c������������������������������������������������������������� conference paper validation of a qualitative behaviour score during the capture phase of stray dogs laura menchetti1, cecilia righi2, gabriella guelfi1, claudia enas1, livia moscati2, stefania mancini3, silvana diverio1* 1 laboratory of ethology and animal welfare (leba), department of veterinary medicine, university of perugia, via san costanzo 4, 00126, perugia, italy 2istituto zooprofilattico sperimentale dell'umbria e delle marche “togo rosati”, via salvemini 5, 06126, perugia, italy 3public veterinary services for urban hygiene and prevention of stray dogs, usl umbria 1, municipal rescue dog shelter, strada per brufa snc, 06148 collestrada (perugia), italy *e-mail: silvana.diverio@unipg.it summary there are no studies on the capture phase of stray dogs, although this can be very stressful and have a significant impact on dogs’ welfare. in this preliminary study, we propose a simple qualitative evaluation system of the dog’s behaviour during the capture phase. the assessments of the animal control officers (aco) were compared with qualitative and quantitative evaluations carried out by two testers to verify their reliability and validity. the agreements and correlation analysis showed that the qualitative score of the testers was reliable and valid. conversely, the scores attributed by the aco were not in agreement and not consistent with the behavioural observations of the testers. these results suggest that the aco did not have the necessary familiarity with behavioural assessments. it should also be considered that the testers made their observations in a different context, and the dog can react differently to different stressors according to his personality and past experiences. in conclusion, the qualitative assessments during the capture phase require implementations and further investigations. keywords: shelter dogs; dog behavior evaluation; animal control officer page 7 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution pet behaviour science | 2019, vol.7, 7-10 doi: 10.21071/pbs.v0i7.11799 conference paper introduction as recognised by the world organisation for animal health (oie, 2015), the process of catching a dog before entering the shelter can be very stressful and have a significant impact on dogs’ welfare. however, as far as we know, there is no study on the behaviour and welfare at the capture time of stray dogs. several researchers have proposed welfare assessment systems in the shelter (hiby, rooney, & bradshaw, 2006; part et al., 2014; protopopova, 2016; rooney, gaines, & bradshaw, 2007) but no one involved the animal control officers (aco). moreover, time constraints and limited staff availability, as well as the complexity of the evaluation form, could represent a real obstacle to the systematic use of welfare assessment tools in dog shelters (barnard et al., 2016; kiddie & collins, 2014). we developed a behaviour assessment system involving different staff figures and consisting of simple and quick qualitative processes. this preliminary study aimed to analyse the validity of the behavioural evaluations carried out by the aco during the capture of stray dogs to introduce it as a new management tool for improving the welfare of dogs entering the shelter. materials and methods after the capture time, the aco had to fill in a form with the demographic data of the dog and assign to his behaviour a score from 0 (extremely calm and sociable) to 5 (extremely stressed; overall, 2013) called stress level. each score was alongside a brief description and a schematic graphic representation of the dog's behaviour. moreover, all aco were trained before the start of the study to recognize sign of stress in the dog. within the first three days of shelter intake, the same dog was evaluated by a tester who compiled the same form and performed behavioural observations of the dog while inside his quarantine pen. the tester recorded the dog’s behaviour by using the scan animal sampling method at 10-second intervals for 15 minutes and a specific ethogram. the percentage of each behavioural variable was calculated within each session. besides, the diversity of behaviour patterns performed was calculated for each dog using the shannon diversity index (h index; part et al., 2014). to evaluate construct validity, we analysed the correlation between stress level (as score) and behavioural observations (as percentage of each behaviour) by using the spearman coefficient (ρ), while we used the cohen’s κ and kendall’s τ tests to evaluate inter-observed reliability. two independent testers evaluated five dogs simultaneously to determine the inter-rater reliability of the stress level. page 8 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution conference paper results and discussion in this work, we analysed the validity of the qualitative behavioural evaluations during the capture of stray dogs by determining their inter-observer agreement and construct validity (meagher, 2009). there was a reasonable inter-observer agreement between the two testers who evaluated the same observational situation (τ=0.490; p<0.001) suggesting that our score was reliable (meagher, 2009). moreover, the associations between the score given by the tester and the behavioural observations were consistent (p<0.05 for 6 ρ coefficients), supporting the construct validity of the qualitative measurement. however, we did not find agreement between the score attributed by the testers and aco. moreover, the score of the aco was not associated in a meaningful way with any behaviour. we hypothesise that the aco are not very familiar with the behavioural assessments and/or that the training received was insufficient. however, we must keep in mind that the assessments were made in different contexts: the capture phase and the pen. moreover, behavioural reactions to an intense stressor depend on many factors, including the dog’s personality and past experience, and the environmental context (hiby et al., 2006; protopopova, 2016; rooney et al., 2007; walker et al., 2016). it is therefore plausible that the same dog reacts differently to different stress stimuli, in relation to different factors. in conclusion, the qualitative assessment during the capture phase requires implementations and further investigations. however, our findings show that the behavioural assessment of shelter dogs can benefit from different points of view. in fact, the shelter staff can monitor the dog in diversified contexts, making more accurate the judgment on his adaptability. references barnard, s., pedernera, c., candeloro, l., ferri, n., velarde, a., & villa, p. d. (2016). development of a new welfare assessment protocol for practical application in long-term dog shelters. veterinary record, 178(1), 18. https://doi.org/10.1136/vr.103336 hiby, e. f., rooney, n. j., & bradshaw, j. w. s. (2006). behavioural and physiological responses of dogs entering re-homing kennels. physiology and behavior, 89(3), 385–391. https://doi.org/10.1016/j.physbeh.2006.07.012 kiddie, j. l., & collins, l. m. (2014). development and validation of a quality of life assessment tool for use in kennelled dogs (canis familiaris). applied animal behaviour science, 158, 57–68. https://doi.org/10.1016/j.applanim.2014.05.008 page 9 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution conference paper meagher, r. k. (2009). observer ratings: validity and value as a tool for animal welfare research. applied animal behaviour science, 119(1–2), 1–14. https://doi.org/10.1016/j.applanim.2009.02.026 oie. (2015). stray dog population control. terrestrial animal health code, 1–17. overall, k. (2013). manual of clinical behavioral medicine for dogs and cats. (e. h. sciences, ed.). saint louis mo: elseviermosby. part, c. e., kiddie, j. l., hayes, w. a., mills, d. s., neville, r. f., morton, d. b., & collins, l. m. (2014). physiological, physical and behavioural changes in dogs (canis familiaris) when kennelled: testing the validity of stress parameters. physiology and behavior. https://doi.org/10.1016/j.physbeh.2014.05.018 porritt, f., shapiro, m., waggoner, p., mitchell, e., thomson, t., nicklin, s., & kacelnik, a. (2015). performance decline by search dogs in repetitive tasks , and mitigation strategies. applied animal behaviour science, 166, 112–122. https://doi.org/10.1016/j.applanim.2015.02.013 protopopova, a. (2016). effects of sheltering on physiology, immune function, behavior, and the welfare of dogs. physiology & behavior, 159, 95–103. https://doi.org/10.1016/j.physbeh.2016.03.020 rooney, n. j., gaines, s. a., & bradshaw, j. w. s. (2007). behavioural and glucocorticoid responses of dogs (canis familiaris) to kennelling: investigating mitigation of stress by prior habituation. physiology and behavior, 92(5), 847–854. https://doi.org/10.1016/j.physbeh.2007.06.011 walker, j. k., dale, a. r., d’eath, r. b., & wemelsfelder, f. (2016). qualitative behaviour assessment of dogs in the shelter and home environment and relationship with quantitative behaviour assessment and physiological responses. applied animal behaviour science, 184, 97– 108. https://doi.org/10.1016/j.applanim.2016.08.012 page 10 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution c������������������������������������������������������������� conference paper marking frequency during intraspecific socialization sessions is related to urinary cortisol levels in shelter dogs daniela alberghina*, gina pumilia, pierluigi raffo, giuseppe distefano, giuseppe piccione and michele panzera department of veterinary sciences, university of messina, italy. *e-mail: dalberghina@unime.it summary the aim of this study was to determine whether behavioural indicators such as marking frequency and whether cortisol/creatinine ratio (c/cr) are influenced by three socialization sessions. six adult shelter dogs were selected: group 1 (n.3) with adequate social experience and group 2 with behavioural problems of aggression against humans (n.2) and excessive fear response towards conspecifics (n.1). spontaneous urine samples were collected for c/cr after socialization sessions. c/cr was positively correlated with urinary marking frequency (p<0.01) and negatively with faecal marking frequency (p<0.05). the correlation with urinary marking could be related to increased release of aldosterone under acth stimulus or to chemical information that dogs are in a stressful situation. the negative correlation with frequency of faecal marking could be related to inhibition of act in a novel situation as stress effect. noninvasive monitoring for detection of cortisol could be useful to assess canine behaviour modulation following intraspecific socialization sessions. keywords: urinary cortisol; shelter dog; marking page 1 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution pet behaviour science | 2019, vol.7, 1 – 6 doi: 10.21071/pbs.v0i7.11798 conference paper introduction examination of social behaviour is very important for screening shelter dogs to be placed for adoption or for identifying dogs that need behavioural intervention. shelter should provide behavioural rehabilitation for fearful and un-socialized dogs in order to increase their chances of adoption. intraspecific socialization should be an important component of behavioural rehabilitation of un-socialized dogs. intraspecific social exposure sessions could be useful to modulate undesirable behaviour in owned dogs (distefano and alberghina, 2016) but also to increase social behaviour and working ability in military dogs (gfrerer et al., 2018). urinary cortisol:creatinine ratio (c/cr) is probably the most widely used physiological indicator, which is reported in published studies about canine welfare (hewson et al., 2007) but individual variability and lack of specificity as a stress response have led researchers to question the value of glucocorticoid levels as a welfare indicator (part et al., 2014). in this study, we hypothesized that social exposure sessions could affect cortisol and behaviours, such as urinary and faecal marking, in shelter dogs. material and methods six adult gonactemizated dogs were chosen from the animal shelter in rovereto (tn), italy. they were all housed in the shelter for six months or more. group 1 (wallace, acab and frida) with adequate social experience and group 2 with behavioural problems of aggression against humans (asia and cloe) and excessive fear response towards conspecifics (baloo)(table 1). each dog of group 2 had the opportunity to have contact with one conspecific of group 1 within the fenced area (10 m×20m) from 12.00 to 12.30 pm. frequency of urination and defecation was monitored. table 1. characteristics of gonactemizated dogs used in the study dog estimated age (years) gender breed group wallace 7 m german sheperd i acab 3 f german sheperd frida 7 f crossbred baloo 7 m hound ii cloe 2 f crossbred asia 9 f argentin mastiff page 2 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution conference paper in a first phase, the experimenter controlled the dog with a leash while direct contact with conspecifics was allowed. in a first phase of social sessions, all dogs wore a baskerville muzzle to ensure that no dog could get harmed. the dog trainer supervised all training and decided which dog wore a muzzle or when a dog could be off the leash. the behaviour of the dogs during social exposure session was video recorded. spontaneously voided urine samples were collected 6 hours after the end of socialization session when subjects were let out of their box. urine was collected by p-scoop dog urine collector and transferred in sterile cups. samples were frozen at -20°c until further analysis for cortisol and creatinine. cortisol in urine was analysed by means of a microtiter plate reader (ez read 400 120 elisa, biochrom, cambridge, united kingdom) with a commercial cortisol urine-elisa kit according to the manufacturer’s instructions (cortisol urine elisa, ldn gmbh & co. kg). urinary creatinine (cr) was also assessed in order to index, and control for, varying urine concentration levels. creatinine assessment was completed with a commercially available kit using a modified jaffe method (giesse diagnostics srl, italy) by means of a uv spectrophotometer (model slim seac, firenze, italy). urinary cortisol, corrected for creatinine, to give urinary cortisol creatinine ratio (c/cr nmol/μmol) was calculated. analysis of variance (anova) with repeated measures was used to assess the influence of sessions on c/cr. bonferroni’s multiple comparison test was applied for post hoc comparisons. pearson’s linear correlation was used to evaluate the relationship between urinary and faecal frequency marking and c/cr. all the statistical analyses were performed using statistica 8 software (statsoft inc., tulsa, ok, usa). p-values <0.05 were considered statistically significant. results levels of c/cr after sessions were significantly influenced by time (f=6.196; p<0.05, fig. 1). a significant positive correlation was found between c/cr and urinary marking frequency (r=0.654, p<0.01 fig.2). a negative correlation was found between c/cr and faecal marking frequency (r=-0.546 p<0.05, fig. 3). page 3 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution conference paper figure 1. influence of social sessions on c/cr levels in six shelter dogs figure 2. linear correlation between c/cr and urinary marking frequency page 4 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution conference paper figure 3. linear correlation between c/cr and fecal marking frequency discussion urine capture is painless, non-invasive, and inexpensive, which is an advantage over other measures gained by invasive techniques (e.g. from serum or plasma). cortisol levels in urine reflect a stress response in the previous 4-8 hours (casey, 2005). levels of cortisol, measure the metabolic rate of the animal and increase during arousal in positive events as well as negative events, therefore, it is possible that the elevated cortisol levels after social exposure were due to arousal and the resulting increase in activity and not necessarily an indication of stress (protopopova, 2016). the small group of socialized dogs showed a tendency to have lower cortisol levels, lower frequency of urination and higher frequency of defecation than the small group of dogs with behavioural problems. the positive correlation between c/cr and frequency of urinary marking could be related to the increased release of aldosterone under acth stimulus or to chemical information that dogs are in a stressful situation. no previous research on canine marking behaviour has evaluated relationship with urine cortisol levels. previously, in relation to acute stress, no significant correlations between behavioural and physiological parameters (salivary cortisol and heart rate) have been observed (beerda et al., 1998). tod et al. (2005) reported that defecation and urination were excluded from analysis of a behavioural test for page 5 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution conference paper efficiency of dog appeasing pheromone, in relation to stress and fear related behaviour due the absence or rarity of their occurrence during testing. the negative correlation between fecal marking frequency and c/cr levels in this study, could indicate that in a novel situation related to intraspecific social exposure, defecation could be inhibited. further studies are necessary to evaluate the role of marking communication between dogs in relation to arousal and stress. references beerda, b., schilder, m. b. h., van hooff, j. a. r. a. m., de vries, h. w., mol, j. a. behavioral, saliva cortisol and heart rate responses to different types of stimuli in dogs. applied animal behaviour science 58 (1998), 365–381. casey r. 2002. fear and stress in companion animals. in: horwitz d, mills d, heath s, editors. bsava manual of canine and feline behavioral medicine. gloucester: br small animal veterinary association; p. 144–153. distefano, g., alberghina, d. moderator dogs in modulation of canine behavioral problems. dog behaviour 2 (2016), 74. gfrerer, n., taborsky, m., würbel, h. benefits of intraspecific social exposure in adult swiss military dogs. applied animal behaviour science 201 (2018), 54-60. hewson,c.j., hiby, e.f. bradshaw, j.w.s. assessing quality of life in companion and kennelled dogs: a critical review. animal welfare 16 (2007), 89-95. part,c.e., kiddie, j.l., hayes, w.a., mills, d.s., neville, r.f., morton, d.b., collins, l.m. physiological, physical and behavioural changes in dogs (canis familiaris) when kennelled: testing the validity of stress parameters. physiology & behavior 133 (2014), 260271. protopopova, a. effects of sheltering on physiology, immune function, behavior, and the welfare of dogs. physiology & behavior 159 (2016), 95–103. tod, e., brander, d., waran, n. efficacy of dog appeasing pheromone in reducing stress and fear related behaviour in shelter dogs. applied animal behaviour science 92 (2005), 295– 308. page 6 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution c������������������������������������������������������������� conference paper proposal of a customized animal welfare protocol for military kennels otavio augusto brioschi soares1*, fernanda ishi2, josé luiz vetorazzo2, felipe borges soares1, nivea de mattos goes vieira1 1brazilian army, 2universidade de sorocaba, brasil. *e-mail: capvetaugusto@gmail.com summary the guarantee of animal welfare has been modernly approached in both physical and emotional aspects. the objective of this work was to propose a management protocol that maximizes animal welfare for working dogs, and that takes into account the particularities of brazilian military institutions, so being able to be implemented more easily. a literature search was conducted and a task force was created to inform the project. after the review, meetings and discussions, the writing of the protocol was finalized. it is divided into eight theoretical and practical prompts, sometimes exemplified. the proposed protocol covers the theoretical proposals on animal welfare found in the literature, in addition to respecting the characteristics and peculiarities of the institutions and military routines in question, which would theoretically facilitate their implementation. after consolidating this proposal, new studies are needed to validate the protocol through its implementation. keywords: working dogs’ welfare, animal welfare, military working dogs page 24 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution pet behaviour science | 2019, vol.7, 24-28 doi: 10.21071/pbs.v0i7.11802 conference paper introduction the guarantee of animal welfare has been modernly approached in both physical and emotional aspects (rooney et al., 2009). working dogs, despite having similar needs as other types of dogs, have particularities in their selection, environment and routine that should be considered (rooney et al., 2005). additionally, animal welfare is decisively influenced by beliefs and values, varying according to the local culture, the nature and the importance that animals assume in different communities (food and agriculture organization of the united nations, 2009). working dogs, and particularly those of military employment, are considered valuable tools by the institutions that employ them (brazilian army, 2013) and companions of the military personnel who have them in their daily lives (lefebvre et al., 2007). within this context, the objective of this work was to propose a management protocol that maximizes animal welfare for working dogs, and that considers the particularities of brazilian military institutions, so being able to be implemented more easily. material and methods a search was made in the national and world literature relating to working animals’ welfare for the theoretical basis of the proposal. a task force, composed of heads of military kennels, trainers and veterinarians, involved in the routine of the kennels’ dogs of the brazilian army in the state of são paulo, was assigned to the task. the group held three face-to-face meetings and virtual discussions to develop the protocol. results after the review, meetings and discussions, the writing of the protocol was finalized. it is divided into eight theoretical and practical points, sometimes exemplified: 1.ensure good quality nutrition and hydration, compatible with the specific individual requirements of the animals, following veterinarian’s advice; 2. ensure preventive and healing medical care, with planned follow-up and previously available veterinary medical assistance compatible with the activity performed; 3. provide bio-climatological adequate daily environments and redouble care when the animals are in other environments, for instance, the supply of transport vehicles for dogs; 4. provide appropriate leading and training equipment that ensure the physical and emotional integrity of the animals, such as the reduced use of choke chain collars and the nonuse of electronic collars without the page 25 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution conference paper assistance of a behavioral veterinarian and/or consultant; 5. to reduce as much as possible the aversive stimulations, especially those of chronic character, by optimizing the use of the critical period of socialization of the puppies, the use of training practices that respect the least invasive minimally aversive (lima) protocol and the use of fear free practices in veterinary medicine; 6. promote routines of physical and cognitive activities that promote motivation and relief of mental and physical stress, such as guided physical training routines and practices of cognitive and food environmental enrichment; 7. promote the technical training of handlers with the concepts of ethology, cognition and animal welfare, through the insertion of hours of this knowledge in the various training courses and continuing education programs offered in the institutions; 8. keep records of the abovementioned behaviors and objective parameters of well-being previously listed, such as periodic stress tests on a sample of animals or individualized monitoring of the animals by a behavioral veterinarian. discussion although today there are several currents that categorize the thought proposals on animal welfare, two deserve to be emphasized: the bio-physiological and the mental/emotional. broadly speaking, such proposals refer to the physical aspects of welfare reflected in the organic development and maintenance of good animal health and the mental aspects of welfare reflected in the sensation of mental satisfaction and the absence of chronic stress disease as being essential (duncan, 2005). within these proposals, points 1 to 3 aim to ensure the physical aspects of welfare and points 4 to 7 seek to work with the mental and emotional animal aspects. according to broom and johnson (1993) there are some aspects that can be taken as parameters for measuring welfare levels such as: expression of a wide range of behaviors considered as normal for the species, conditions and characteristics of presentation of preferred behaviors, and analysis of the physiological indicators of pleasure. at that matter, point 8, the one that concerns welfare measures and registration, aims at the production of information that will allow planning, re-evaluation and possible redirection of actions, besides being an objective measurement of the effectiveness of the protocol itself. in this way, each institution or military unit must determine the most appropriate indicators for monitoring the implementation of the protocol. the maintenance of management and training techniques considered as traditional, based on principles that have sometimes been overcome, and without proper evaluation of the page 26 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution conference paper negative implications of non-management of aversive stimuli, may represent, through increased stress levels, a possible risk to emotional health (lieberman, 1999). in this way, there was an intense concern on the part of the working group, demonstrated in prompts 5, 6 and 7, to advocate modern training and behavior modification techniques (international association of animal behavior consultants, 2018) and avoid techniques that may compromise animal welfare and cause behavioral problems, such as choke chain and electronic collars (coopers & mills, 2014). these items become of paramount importance for the military routine, since dogs with low welfare levels may lose explosive detection work performance (rooney et al., 2005). the proposed protocol covers the theoretical proposals on animal welfare found in the literature, in addition to respecting the characteristics and peculiarities of the institutions and military routines in question, which would theoretically facilitate their implementation. after consolidating this proposal, future research is needed to validate the protocol through its implementation references cooper, j.j., cracknell, n., hardiman, j., wright, h., & mills, d. (2014). the welfare consequences and efficacy of training pet dogs with remote electronic training collars in comparison to reward based training. plos one, 9(9), e102722. duncan, i.j. (2005). science-based assessment of animal welfare: farm animals. rev. sci. tech. off. int. epiz., 24(2). 483-492. exército brasileiro (2013). caderno de instrução de emprego de cão de guerra (eb70-ci11.002). 51p. food and agriculture organization of the united nations. (2009). capacity building to implement good animal welfare practices. rome. 80p. international association of animal behavior consultants. (2018). iaabc position statement on lima. retrieved september 1, 2011 from https://m.iaabc.org/about/positionstatements/lima/ page 27 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution conference paper lefebvre. d., diederich, c., delcourt, m. & giffroy, j.m. (2007). the quality of the relation between handler and military dogs influences efficiency and welfare of dogs. applied animal behaviour science, 104, 49–60. lieberman, d. (1999). learning: behavior and cognition. london: wadsworth. rooney, n., bradshaw, j.w.s. & almey, h. (2004). attributes of specialist search dogs—a questionnaire survey of uk dog handlers and trainers. journal of forensic science 49(2), 300-306. rooney, n., gaines, s. & hiby, e. (2009). a practionerś guide to working dog welfare. journal of veterinary behavior 4, 127-134. page 28 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution c������������������������������������������������������������� conference paper cannabidiol as a potential anti-epileptic dietary supplement in dogs with suspected epilepsy: three case reports chie mogi* and takaaki fukuyama yamazaki university of animal health technology, japan e-mail:chiemogi@gmail.com summary epilepsy is the most common chronic neurological disorder in dogs and the almost lifelong administration of anti-epileptic drugs (aeds) is recognized as the most successful treatment in veterinary medicine. current pharmacological therapies for epilepsy have shown undesirable side effects. the dietary use of cannabidiol (cbd) in humans has shown therapeutic potential for the treatment of epileptic seizures. we administered cbd for 8 weeks to three dogs with epileptic seizures; decrease in the seizure interval was observed in two dogs, while one dog showed no improvement. regarding the owners’ impressions, one reported considerable symptom improvement, one that the symptoms improved, and one that the symptoms remained unchanged. keywords: keyword 1, cannabidiol; keyword 2, dog; keyword 3, epilepsy page 11 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution pet behaviour science | 2019, vol.7, 11 –16 doi: 10.21071/pbs.v0i7.11800 conference paper introduction epilepsy is the most common chronic neurological disorder in dogs (heske et al, 2013), and the almost lifelong administration of anti-epileptic drugs (aeds) is recognized as the most successful treatment in veterinary medicine (de risio et al, 2015). however, many potential adverse effects of aeds, including polyphagia, sedation, restlessness, and polydipsia, have been reported (charalambous et al., 2016). to achieve effective seizure control without reducing the quality of life of dogs with epilepsy, new treatment strategies are needed (martlé et al, 2014). in humans, the dietary use of cannabidiol (cbd) is reported to reduce seizure frequency and duration (devinsky et al, 2016); the serum levels of commonly used aeds also increase with increasing cbd dose (gaston et al, 2017). cbd, a phytocannabinoid compound derived from the cannabis plant, has been gaining attention as a potential anticonvulsant without the psychoactive effects seen with tetrahydrocannabinol (gaston et al, 2017); however, these beneficial therapeutic effects have not yet been explored in dogs with epilepsy. therefore, we conducted an open-label cbd study in three dogs with epileptic seizures to investigate the efficacy, tolerability, and safety of cbd. case description clinical and therapeutic history of dog 1: the dog was a 3-year 2-month-old, 33-kg castrated male labrador retriever raised with 5 other dogs. the owner first noted episodes of seizure-like behavior when the dog was about 6 months old. during these episodes, the dog lay shivering on the floor with his head up; these bouts generally lasted several minutes. the dog had not received any aeds. he had experienced 6 episodes this year, with an interval of approximately 30 days. clinical and therapeutic history of dog 2: the dog was a 11-year 2-month-old, 4-kg castrated male papillon raised with 3 other dogs. the owner first noted episodes of epilepsy when the dog was 3-years, 11 months old. zonisamide (consave, ds pharma animal health co., ltd., japan) 15 mg/kg, an aed, was administered twice daily for the last three years. the attacks were spaced about 2-3 months apart. during the cbd treatment period, the dog received only zonisamide 15 mg/kg twice daily. he also had urinary incontinence and salivation (foaming) as a result of the attacks. page 12 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution conference paper clinical and therapeutic history of dog 3: the dog was a 10-year 2-month-old, 2-kg male chihuahua raised alone by a family containing children. the owner first noted episodes of epilepsy when the dog was 3 years old. during these episodes, the dog lay on the floor shivering and salivating. he had experienced 2 episodes this year, and the intervals were irregular. table 1. cases in the study case age years se body weightkg breed cbd trade name dosage (mg/kg/day) aed dog1 2 m 33 labrador retrieve r hemp oil 1700 0.51 none dog 2 10 m 4 papillon hemp oil 1000 (for first 5 weeks) 1.25 zonisamide hemp oil 330 (for remaining 3 weeks) 1.24 zonisamide dog 3 9 m 2 chihuahua hemp oil 1000 5.00 none table 2. seizure interval. seizure dates during the administration period are boldface. case cbd administration period seizure date seizure interval (days) owners’ impressions dog 1 2018/6/26 2018/8/21 2018/6/14 improved 2018/8/8 55 2018/8/11 3 2018/8/26 15 2018/9/3 8 dog 2 2018/6/18 2018/8/14 2018/4/22 unchanged 2018/6/26 65 2018/6/27 1 2018/6/28 1 2018/6/29 1 2018/8/10 42 2018/8/11 1 2018/8/12 1 2018/8/13 1 dog 3 2018/6/11 2018/8/10 2018/5/5 considerably improved 2018/5/8 3 　 　 2018/7/11 64 　 page 13 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution conference paper treatment each dog received a plant-derived formulation of natural cbd-containing full-spectrum hemp extract, pet releaf hemp oil (peat releaf ltd, u.s.a.), in organic coconut oil. cbd was administered in two divided doses (12 hourly) on an empty stomach. the efficacy, tolerability, and safety of cbd were assessed every 2 weeks for 8 weeks. outcomes of cbd treatment and owner perception (dog 1): the male labrador retriever received cbd at 0.51 mg/kg/day. the dog had 2 epileptic attacks during the 8-week treatment course. after withdrawal of cbd, an epileptic attack occurred within 5 days. the owner reported that the dog slept longer and barked less in the daytime, even when other dogs were excited, during the first 2 weeks than in the preceding weeks. overall, the owner felt that the dog showed improvement. outcomes of cbd treatment and owner perception (dog 2): the male papillon received cbd at 1.24-1.25 mg/kg/day. the dog had 8 epileptic attacks during the 8-week treatment course. during the first 2 weeks, the owner reported that the dog ate more willingly. it was also noted that the dogs settled down and slept longer during the day. overall, the owner’s impression was unchanged. outcomes of cbd treatment and owner perception (dog 3): the male chihuahua received cbd at 5.00 mg/kg/day. the dog had only 1 attack during the 8-week treatment course. the owner felt that seizure-like behavior during the attacks had decreased slightly with treatment. the owner also reported that the dog showed less aggression toward familiar people, such as the owner’s children. this study was approved by the committee for institutional animal care and use of yamazaki university of animal health technology (permission number: 20180630-002) and carried out according to the guidelines of the committee. discussion in this study, the seizure frequency improved considerably and owners reported a positive impression. in humans, the mental factors implicated in epileptic seizures have been successfully treated with behavioral interventions (martinovic, 2001). mental stress, such as anxiety, can influence seizure episodes. blessing et al. reported that cbd has considerable potential for treatment of multiple anxiety disorders in patients with epilepsy page 14 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution conference paper (blessing et al., 2015). this anxiolytic effect may attenuate the symptoms of epilepsy in dogs as well as humans. further research is needed for better understanding the neurobiological mechanisms of cbd treatment. this study has several limitations. first, the number of dogs was small and the owners may not be representative of all epileptic dog owners. dogs with more severe epilepsy phenotypes may be administered aeds. to investigate the potential interactions of cbd with aeds, further research is needed. acknowledgment takakura new industries inc. provided the study dietary supplement and collected and collated the data from all sites. toru kimura, dvm, minami koganei animal hospital, kindly provided clinical and therapeutic information of dog 2. references blessing, e. m., steenkamp, m. m., manzanares, j., & marmar, c. r. (2015). cannabidiol as a potential treatment for anxiety disorders. neurotherapeutics, 12(4), 825-836. charalambous, m., shivapour, s. k., brodbelt, d. c., & volk, h. a. (2016). antiepileptic drugs’ tolerability and safety–a systematic review and meta-analysis of adverse effects in dogs. bmc veterinary research, 12(1), 79. de risio, l., bhatti, s., muñana, k., penderis, j., stein, v., tipold, a., ... & mandigers, p. j. (2015). international veterinary epilepsy task force consensus proposal: diagnostic approach to epilepsy in dogs. bmc veterinary research, 11(1), 148. devinsky, o., marsh, e., friedman, d., thiele, e., laux, l., sullivan, j., ... & wong, m. (2016). cannabidiol in patients with treatment-resistant epilepsy: an open-label interventional trial. the lancet neurology, 15(3), 270-278. gaston, t. e., bebin, e. m., cutter, g. r., liu, y., szaflarski, j. p., & uab cbd program. (2017). interactions between cannabidiol and commonly used antiepileptic drugs. epilepsia, 58(9), 1586-1592. heske, l., nødtvedt, a., jäderlund, k. h., berendt, m., & egenvall, a. (2014). a cohort study of epilepsy among 665,000 insured dogs: incidence, mortality and survival after diagnosis. the veterinary journal, 202(3), 471-476. page 15 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution conference paper martinovic, ž. (2001). adjunctive behavioural treatment in adolescents and young adults with juvenile myoclonic epilepsy. seizure european journal of epilepsy, 10(1), 42-47. martlé, v., van ham, l., raedt, r., vonck, k., boon, p., & bhatti, s. (2014). nonpharmacological treatment options for refractory epilepsy: an overview of human treatment modalities and their potential utility in dogs. the veterinary journal, 199(3), 332339. page 16 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution c������������������������������������������������������������� conference paper heart rate, heart rate variability and salivary cortisol as indicators of arousal and synchrony in clients with intellectual disability, horses and therapist during equine-assisted interventions anna naber1, lena kreuzer2, roswitha zink1, eva millesi3, rupert palme4, karin hediger5,6, lisa maria glenk2* 1 e.motion equotherapy, vienna, austria 2 comparative medicine, the interuniversity messerli research institute of the university of veterinary medicine vienna, medical university vienna and university vienna, austria 3 department of behavioural biology, university of vienna, austria 4 unit of physiology, pathophysiology und experimental endocrinology, university of veterinary medicine vienna, austria 5 faculty of psychology, university of basel, switzerland 6 swiss tropical and public health institute, department of epidemiology and public health, switzerland *e-mail: lisa.glenk@vetmeduni.ac.at summary the aim of the study was to analyse interaction processes in equine-assisted therapy (eat) sessions with ten female clients in the period of emerging adulthood with intellectual disability (id). heart rate (hr), heart rate variability and salivary cortisol levels have been analysed in humans and horses before, during and after a standardised therapy session as well as in a control condition. there was a trend of lower cortisol levels and higher variability and parasympathetic tone induced by horses. during challenge however, there was a significant lower hr in the horse condition. significant correlations in heart rate between therapist, client and horse were found with stronger interaction with a familiar horse. our findings suggest that eat may effectively modulate stress in humans with id. our results further elucidate synchronisation patterns in hr highlighting the pivotal role of relationship quality and intensity as modulators of synchrony. keywords: heart rate variability, equine-assisted therapy, cortisol page 17 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution pet behaviour science | 2019, vol.7, 17-23 doi: 10.21071/pbs.v0i7.11801 conference paper introduction a growing body of research has attributed effects of human-horse interaction to improved bio-psycho-social health. intellectual disability (id) refers to impaired intellectual and adaptive functioning and equine-assisted therapy (eat) has been considered a promising practice as complementary treatment of id (e.g. borgi et al. 2016), however its feasibility with regard to stress reduction has been questioned (anestis et al. 2014). while evidence on physiological benefits such as increased motor control exists (del rosario-montejo et al. 2015), psychological benefits and underlying mechanisms of eat are not yet fully settled (kendall et al. 2014). effective autonomic and adrenal regulation of arousal plays a keyrole in the maintenance of (mental) health and progression of diseases (glenk & kothgassner 2017). coordination of nonverbal behaviors between interactive partners takes place during the process of synchronization in many mammalian species. the experience of synchrony roots in the mother-child relationship, and high levels of synchrony have been related to efficient bonding and stress reduction (aztil et al. 2014; leclère et al. 2014). interactions with a high level of synchronization are more efficient, also across species (julius et al. 2014, pirrone et al. 2017). this study sought to elucidate mechanisms and psychological effects of eat within the therapeutic triangle (i.e. participating humans and horses, see figure 1). figure 1. therapeutic triangle in eat. arrows indicate bi-directional relationships between client, horse and therapist. page 18 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution conference paper the aims of this study were: 1) to investigate physiological effects of a standardized eat for young adults with id 2) to explore synchronization patterns between humans and animals material and methods ten eat-experienced women (mn = 21.8, sd = 3.39) with mild (n=5) to moderate (n=5) id according to the glasgow level of ability and developement scale (cooray et al., 2015) participated in the study and were paired with their favourite (n=5) or an unfamilar horse (n=5). the therapist (n=1) took part with familiar (n=2) and less familiar horses (n=2). each client underwent two eat sessions with horse and two control sessions with the same schedule (baseline, relaxation, challenge) but where on-horse activities were carried out on a barrel horse (see table 1). during relaxation phases, therapist-guided recreation exercises were carried out, while the challenge phase included a simple cognitive-motor task. hr was recorded with a polar v800 telemetric system. objective measures of autonomic activation were heart rate (hr) and heart rate variability (hrv), i.e. overall variability (sdnn); sympathicovagal balance (lf/hf ratio) and parasmypathetic tone (rmssd, sd1). salivary cortisol served as an indicator for adrenocortical stimulation and was measured prior to (s1), during (s2) and after (s3, s4) eat. table 1. study design. indicates 5 min intervals of hr recordings; bl…baseline, ch… challenge, r…relaxation, s1-s4….saliva samples page 19 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution conference paper as shown in figure 2, hr of clients during challenge was significantly lower under the horse condition compared to the control (t(19) = -5.569; p<0.01). relaxation phase 2 and baseline 2 indicated a trend toward lower hr with horse. no differences in salivary cortisol emerged between the conditions (x2 (15) = 2.13, p = 0.344) except for a non-significant trend toward lower cortisol levels after eat (mn = 4.74) compard to control (mn = 6.59). rmssd of clients tended to be higher in interaction with a horse compared to the control sessions (see table 2). the same trend was found also for sd1 and sdnn, however not for lf/hf ratio. figure 2. mean (±sd) heart rate of clients over the course of a session compared between conditions. * indicates p<0.01 page 20 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution conference paper table 2. rmssd values of clients over the course of a session compared between conditions e (experimental) and c (control). the quality of bond between humans and the therapy horse influenced the hr of the therapeutic triangle. patterns of synchrony regarding mean hr were found for clients who interacted with their favourite horse (rs=.38, p<0.05) but not for clients who interacted with an unknown horse (see figure 3). a stronger correlation of mean hr was found between the therapist and her own horse (rs=.70, p<0.05) compared to a less familiar horse (rs=.55, p<0.05). the hr of horses appeared to be most stimulated in interaction with familiar humans. figure 3. correlation of mean heart rate (clients and horses). page 21 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution p z conference paper discussion our findings suggest that eat may effectively modulate stress in humans with id. despite the small sample, our data revealed that the horse condition led to decreased hr during a challenging cognitive-motor task. the trend of data in overall autonomic variability (sdnn) and parasympathetic tone (rmssd, sd1) point at a stress reduction. interaction with horses may help people with id mobilize resources and feel more understood without needing words for communication. our results further elucidate synchronization patterns in hr highlighting the pivotal role of relationship quality and intensity as modulators of synchrony in both, horses and people. references anestis md, anestis jc, zawilinski ll (2014). equine-related treatments for mental disorders lack empirical support: a systematic review of empirical investigations. j clin psychol 70 (12):1115-1132atzil, s., hendler, t., & feldman, r. (2014). the brain basis of social synchrony. scan, 9, 1193-1202. borgi, m., loliva, d., cerino, s., chiarotti, f., venerosi, a., bramini, m., nonnis, e., marcelli, m., vinti, c., de santis, c. (2016). effectiveness of a standardized equine-assisted therapy program for children with autism spectrum disorder. journal of autism and developmental disorders 46, 1–9 cooray, s., weber, g., alexander, r., roy, a., bhaumik, s., devapriam, j., …, & cooper, s. a. (2015). towards global screening for disorders of intellectual development (did) – the clinical utility of glasgow level of ability & development scale (glads). in … (chair), symposium conducted at the meeting of the european as-sociation of mental health in intellectual disability, florence del rosario-montejo o, molina-rueda f, muñoz-lasa s, alguacil-diego im (2015). effectiveness of equine therapy in children with psychomotor impairment. neurologia. 30(7):425-32 glenk, l. m., & kothgassner, o. d. (2017). life out of balance: stress-related disorders in animals and humans. in e. jensen-jarolim (hrsg.), comparative medicine: disorders linking humans with their animals (s. 97-107). cham: springer page 22 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution conference paper kendall, e., maujean, a., pepping, c. a., & wright, j. j. (2014). hypotheses about the psychological benefits of horses. explore, 10, 81-87 leclère, c., viaux, s., avril, m., achard, c., chetouani, m., missonnier, s., & cohen, d. (2014). why synchrony matters during mother-child interactions: a systematic re-view. plos one, 9, 1-34 julius, h., beetz, a., kotrschal, k., turner, d. c., & uvnäs-moberg, k. (2014). bindung zu tieren. psychologische und neurobiologische grundlagen tiergestützter interventionen. göttingen: hogrefe pirrone, f., ripamonti, a., garoni, e.c., stradiotti, s., albertini m. (2017). measuring social synchrony and stress in the handler-dog dyad during animal-assisted activities: a pilot study . journal of veterinary behavior, 21, 45-52 page 23 2018 open conference | pet behaviour science creative common license 4.0 – non commercial – share alike – attribution the effect of age on visuo-spatial short-term memory in family dogs the effect of age on visuo-spatial short-term memory in family dogs patrizia piotti*, dóra szabó, lisa wallis, zsófia bognár, bianka stiegmann, anna egerer, pauline marty, enikő kubinyi pet behaviour science | 2017, vol.4, 17 – 19 doi: 10.21071/pbs.v0i4.10130 patrizia piotti, dóra szabó, lisa wallis, zsófia bognár, bianka stiegmann, anna egerer, pauline marty, and enikő kubinyi elte eötvös loránd university, pázmány péter sétány 1/c, 1117 short communication * email: patrizia.piotti@yahoo.it budapest, hungary keywords: dog, cognitive ageing, visuo-spatial memory this paper is based on a communication presented at ‘2017 open conference | pet behaviour science’ by the authors. please, visit https://goo.gl/6y7vjd for the prezi presentation introduction ageing in dogs is associated with the decline of several cognitive domains, such as learning, memory, visuospatial function, executive function, and attention (folstein et al. 1975; head 2014; landsberg et al. 2012; mongillo et al. 2013; svicero & amorim 2017; wallis et al. 2014). the visuo-spatial memory domain is particularly interesting, because its decline precedes the onset of declines in other domains. therefore, visuo-spatial memory decline may be an early marker of ageing (head et al. 1995). furthermore, the capacity to acquire and recall the spatial features of a novel location is critical for adaptation to the environment, and impaired spatial ability can have a great impact on quality of life with advancing age. however, previous research on visuo-spatial function decline in dogs are affected by several limitations e.g. 1) testing required extensive training and lengthy protocols; 2) lack of control for potentially confounding medical conditions, such as sensory-motor impairment; 3) subject selection was limited to the laboratory dog population, which differs from non-laboratory dogs in behaviour patterns and response to social stimuli; 4) low sample sizes page 17creative common license 4.0 – non commercial – share alike – attribution 2017 | vol. 4 | 17 19 abstract decline in the visuo-spatial memory domain may be an early marker for cognitive decline and has a relevant impact on animal welfare. current research on visuo-spatial memory in family dogs is often limited by factors such as the need of extensive pre-training, limited attention to co-occurring medical conditions, a focus on laboratory dogs, or low sample size. therefore, we aimed to develop a test that relies on visuospatial short-term memory, may be performed in a short time, and does not require explicit training. we tested a large sample of young and old dogs, finding that young dogs were more likely to perform correctly, although performance decreased with consecutive trials in both age groups. however, groups did not vary in the severity of mistakes. this task represents the first measure of dogs’ age-related decline of short-term spatial memory that does not require explicit training. the test could potentially be used in veterinary behaviour contexts to monitor cognitive changes in ageing dogs, utilizing a simple binary measure of success. https://goo.gl/6y7vjd http://www.petbehaviourscience.org/ (halmágyi 2010; lazarowski & dorman 2015; szabó et al. 2016; wallis et al. 2014). for these reasons, the use of existing paradigms is limited to laboratory settings, and provides little benefit to the non-laboratory canine population. we therefore designed a novel task (part of a larger battery of tests), which did not require any explicit training of the dog and could be performed in the time span of a few minutes, and tested a large sample of non-laboratory dogs. methods we were interested in measuring whether the task could detect age-dependent short-term memory changes, thus, we compared two groups of dogs, based on their age; ‘young dogs’ (n = 44, mdnage = 4.6 years, iqr = 3.3 – 6.0, females = 61%), and ‘old dogs’ (n = 75, mdnage = 10.7 years, iqr = 9.0 – 11.6, females = 43%). dogs of various breed types, medium to large sized, were recruited through an online survey. before testing, the dogs’ sensory-motor function was assessed in a standardised way, in order to exclude conditions that could potentially impair the dogs’ performance during testing. the tests were performed in an empty room, where 5 identical containers were positioned on the floor equidistant from each other at regular intervals along a semi-circle so that they were all equally distant from a pre-determined location 2 meters away. at the beginning of each trial, the owner stood at the predetermined location and held the dog by the leash. the dog witnessed an experimenter baiting 1 of the 5 identical plastic containers and was then walked out of the room. after a 30 seconds distraction task (petting or playing with the dog), the dog was walked back into the room, was let off leash and was allowed to approach the containers. the experimenter recorded the first choice made by the dog and the severity of mistakes the dog made. the test was repeated once per container; the order of baited locations was counterbalanced across participants. results two linear models were calculated to analyse the effect of age group, breed, and sex on the measured variables. for each model, an automated model selection process generated multiple models with combinations of these factors; models estimating both intercept and/or slope for random effects were also calculated. for each response variable, the model that had the lowest akaike information criterion (aic) score, as tested by likelihood ratio test, was then selected. a generalised linear mixed model (glmm) fit with maximum likelihood approximation, with binomial error structure and logit link function was selected for the response variable ‘first approach’ (correct vs incorrect), with the random intercept factor ‘dog identity’. a main effect without interaction of the fixed factors ‘age group’ (young or old) and ‘trial number’ (1 to 5) on the outcome variable was observed (glmmagegroup+trial(dog), aic= 745.04, n = 595, number of subjects = 119, χ4=14.66, p=0.001). young dogs were more likely to choose the correct container compared to the old dogs (post-hoc tukey: estimateyoung-old ± se = 0.627 ± 0.222, p=0.005), and dogs in both groups were overall less likely to choose the correct container as trials progressed (estimatetrial ± se = 0.165 ± 0.06, p=0.011). for the response variable ‘mistakes’ (scores from 0 to 3, with 0=most severe mistakes; 3=no mistakes), the lowest aic was yielded by another glmm estimated by maximum likelihood with poisson error structure and log link function, with the nested random intercept factors ‘dog identity’ and ‘trial’ and a main effect for the fixed factor ‘age group’ . however, the model was not significantly different from the null model, i.e. including the intercept only (glmmagegroup, aic=1851.2, n=595; number of subjects=119, χ4 = 2.24, p = 0.135). conclusion this task represents the first example of a protocol relying on short-term spatial memory that does not require any explicit training and identifies a difference in performance associated with age. due to its simplicity, the task could potentially be used outside the laboratory environment, e.g. by veterinary professionals, in order to monitor cognitive changes in ageing dogs. care is needed in the selection of the outcome variable: a binary measure might be more effective than scores based on incorrect choices. further research is required to determine performance ranges at the population level and identify changes associated with pathological conditions affecting cognitive abilities, e.g. canine cognitive dysfunction syndrome. page 18 www pet behaviour science org creative commons license 4.0 – non commercial – share alike – attribution 2017 | vol.3 | 17 19 funding this project has received funding from the european research council (erc) under the european union’s horizon 2020 research and innovation programme (grant agreement no. 680040) and was supported by the jános bolyai research scholarship of the hungarian academy of sciences for ek. acknowledgments we would like to thant sarolta marosi, vivien hemző, szandy deés, frida katona, for their help with the data collection and coding and louis le nézet for his help with coding. references folstein, m. f., folstein, s. e., & mchugh, p. r. (1975). a practical state method for. journal of psychiatric research, 12(3), 189–198. http://doi.org/10.1016/00223956(75)90026-6 halmágyi, e. (2010). családi és laboratóriumi beagle kutyák személyiségének összehasonlító elemzése [comparative analysis of the personality of family and laboratory beagle dogs] msc thesis. eötvös loránd university. head, e. (2014). a canine model of human aging and alzheimer’s disease. biochimica et biophysica acta, 1832(9), 1384–1389. http://doi.org/10.1016/j.bbadis.2013.03.016.a head, e., mehta, r., hartley, j., kameka, m., cummings, b. j., cotman, c. w., … milgram, n. w. (1995). spatial learning and memory as a function of age in the dog. behavioral neuroscience, 109(5), 851– 858. http://doi.org/10.1037/0735-7044.109.5.851 landsberg, g. m., nichol, j., & araujo, j. a. (2012). cognitive dysfunction syndrome. a disease of canine and feline brain aging. veterinary clinics of north america small animal practice, 42(4), 749–768. http://doi.org/10.1016/j.cvsm.2012.04.003 lazarowski, l., & dorman, d. c. (2015). a comparison of pet and purpose-bred research dog (canis familiaris) performance on human-guided object-choice tasks. behavioural processes, 110, 60–67. http://doi.org/10.1016/j.beproc.2014.09.021 mongillo, p., araujo, j. a., pitteri, e., carnier, p., adamelli, s., regolin, l., & marinelli, l. (2013). spatial reversal learning is impaired by age in pet dogs. age, 35(6), 2273–2282. http://doi.org/10.1007/s11357-0139524-0 svicero, d. j., & amorim, r. (2017). prevalence of behavioral changes in senile dogs. clinic and surgery, 47(2). http://doi.org/10.1590/0103-8478cr20151645 szabó, d., gee, n. r., & miklósi, a. (2016). natural or pathologic? discrepancies in the study of behavioral and cognitive signs in aging family dogs. journal of veterinary behavior: clinical applications and research, 11, 86–98. http://doi.org/10.1016/j.jveb.2015.08.003 wallis, l. j., range, f., müller, c. a., serisier, s., huber, l., & virányi, z. (2014). lifespan development of attentiveness in domestic dogs: drawing parallels with humans. frontiers in psychology, 5(71), 1–13. http://doi.org/10.3389/fpsyg.2014.00071 page 19 visuo-spatial memory in ageing dogs creative commons license 4.0 – non commercial – share alike – attribution piotti et al. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science influence of low stress handling during clinical visit on physiological and behavioural indicators in adult dogs: a preliminary study influence of low stress handling during clinical visit on physiological and behavioural indicators in adult dogs: a preliminary study bruno scalia, daniela alberghina and michele panzera pet behaviour science | 2017, vol.4, 20 – 22 doi: 10.21071/pbs.v0i4.10131 bruno scalia, daniela alberghina and michele panzera department of veterinary sciences, polo universitario annunziata 98168, university of messina short communication * email: bru.scalia@hotmail.it italy keywords: dog; low-stress handling; veterinary hospital; animal welfare this paper is based on a communication presented at ‘2017 open conference | pet behaviour science’ by the authors please, visit http://bit.ly/scalia2017 to see the poster introduction low stress handling techniques or “fear free principles” in veterinary clinics are becoming an important research area aimed at improving small animal welfare (yin, 2009; overall, 2013; lloyd 2017). if an animal experiences inadequate handling at the veterinary hospital, it is likely to become more fearful and difficult to handle during its next visits due to the poor association between the experience and the environment/personnel (classical conditioning) (lloyd 2017). in a previous study 78.5% of dogs exhibited fear reactions during clinical visits, particularly on the examination table; those with only positive previous experiences were significantly less 'fearful' than those who had previously had a negative experience (döring et al. 2009). increased locomotor activity, panting, lip page 20creative common license 4.0 – non commercial – share alike – attribution 2017 | vol. 4 | 20 22 abstract low stress handling techniques or “fear free principles” in veterinary clinics are becoming an important research area aimed at improving small animal welfare, considering that the majority of dogs who undergo clinical examinations exhibit fear or anxiety signs. objective of this study was to compare a number of physiological and behavioural indicators using low stress handling (lsh) and traditional (tt) techniques in order to assess whether the lsh approach had a positive impact on the dog’s welfare. eight adult dogs were filmed while undergoing both lsh and tt visits (separated by a distance of seven weeks). the same usual sequence of events was followed for both visits (e. g. muzzle wearing, heart and lungs stethoscope examination, etc.) except that 1) during the lsh visit, the dog was free to explore the environment (while receiving treats) and play for five minutes before and after the visit 2) throughout the medical examination the veterinarians’ attitude and handling techniques were always aimed at preventing stress and guaranteeing the best physical support possible. the videos were then evaluated for the number of fear and stress signs the subjects showed. the examined physiological variables were respiration (breath/min), heart rate (hr) and rectal temperature (rt). physiological variables were analysed by t-test for paired data while frequency of behavioural fear indicators by wilcoxon signed-rank test. rectal temperature was within range in both groups but significantly higher (p<0.05) during lsh visit, while low head, lip licks and whale eye behaviours were significantly higher (p<0.05) during tt visit. these results suggest that low stress handling decreases frequency of some fear-related behaviours and could improve the quality of human-dog interactions. future research that aims to replicate and further investigate these results in a large canine population is required. http://bit.ly/scalia2017 http://www.petbehaviourscience.org/ licking, tongue flicking, yawning, paw lifting, shaking, vocalization and lowered body posture have been identified as potential indicators of stress in dogs (csoltova et al. 2017). yin (2009) wrote a useful book to help veterinary staff take a low stress approach to companion animal care in the veterinary hospital. we hypothesized that physiological and behavioural stress indicators in dogs would increase more during traditional clinical examination than during “low stress handling” examination. objective of this study was to compare a number of physiological and behavioural indicators using low stress handling techniques and traditional techniques. methods eight adult dogs (n.3 females and n. 5 males, aged from 3 to 10 years), used for clinical educational purposes at the department of animal medicine and surgery of the university of x, were enrolled in this study. data were collected from march to may 2017. all dogs received routine healthcare and had been previously declared healthy by veterinarians. moreover, they were unfamiliar with the examination room. to reduce the influence of individual variations, each dog was assigned to both the experimental and the control group (separated by a distance of 50 ± 2 days), thus all dogs in this study acted as their own controls. group a (n=4) was assigned to the traditional technique (tt) at the first visit and to low stress handling (lsh) technique during the second visit, whereas group b (n=4) was assigned to lsh at the first visit and to tt during the second visit. the subjects were randomly allocated to either group. the testing procedures were always carried out by two veterinary medicine male students in the role of the veterinarian. the study consisted of two clinical visits: tt: the dog was kept on the examination table, under stationary conditions, using any required restraint. a standardized sequence of examination steps was performed: muzzle wearing, heart and lungs stethoscope examination, muzzle removal; eyes, ears and oral cavity examination; paws inspection; rectal temperature measurement; lateral recumbency positioning and abdominal area inspection; positioning the dog into a sit and simulated jugular venipuncture; sternal recumbency positioning and simulated cephalic vein catheter placement; simulated saphenous venipuncture. throughout the examination, behaviours and heart rate (hr) of the dog were recorded. lsh: before the lsh clinical visit the dog was free to explore the environment for 5 minutes, while the researchers watched the dog's body language, avoided direct eye contact and greeted it correctly; some treats were tossed on the floor. when no signs of fear were detected, the researcher approached the dog in order to begin the low stress handling visit. a soft towel had been laid on the table and tasty treats were available. the sequence of the examination was the same as for the tt visit, except that the veterinarians’ attitude and the handling techniques were aimed at preventing fear and stress and guaranteeing the best physical support possible. after the visit the dog was free to further investigate the environment, received some other treats and was invited to playing activities for approximately 5 minutes. the examined physiological variables were respiration (breath/min), heart rate (hr) and rectal temperature (rt). rectal measurements were taken with a digital thermometer. the thermometer was gently inserted into the rectum for a length of about 2 cm. the thermometer emitted an acoustic signal when the attained temperature remained stable, after approximately 1 minute. the thermometer was disinfected after each sampling. seven behaviours (“low head position”, “panting”, “lip licks”, “yawns”, “whale eye”, “hypervigilance”, “urination and defecation”) were logged as the number of events per 15 min. all behavioural data was collected by video recording. physiological variables were analysed by ttest for paired data while frequency of behavioural fear indicators by wilcoxon signed-rank test using statistica 8 software (statsoft inc., tulsa, ok, usa). the significance level was set at p<0.05. results a significant difference was found between groups on rectal temperature (t=2.942 df=7, p<0.05), low head (w=21, p<0.05), lip licks (w=21, p<0.05) and whale eye behaviours (w=36, p<0.05). rectal temperature was within range in both groups but significantly higher page 21 low stress handling during clinical visit in dogs creative commons license 4.0 – non commercial – share alike – attribution scalia et al. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science during lsh visit than during tt visit (38.78 ± 0.33°c and 38.45 ±0.37°c respectively), while low head, lip licks and whale eye behaviours were significantly higher during tt visit. conclusions the results of this preliminary study indicate that “low stress handling” significantly affected rectal temperature and low head posture, lip licks and whale eye behaviours. the influence on rectal temperature could be related to a different temperature perception through the examination table (lower table temperature during tt than during lsh visits) but many conditions, including digestion, peristaltic movements, fecal masses and physical activity may affect rectal temperature (rexroat et al. 1999; rizzo et al. 2017). lip licking has been previously related to salivary cortisol concentrations in hospitalized dogs (hekman et al. 2012) and could be useful for the evaluation of acute stress levels in a social context (beerda et al. 1998). limitations to the findings of this study are the small number of subjects, the adult age of enrolled dogs and the absence of interaction with an owner. future research that aims to replicate and further investigate these results in a large canine population is required. in conclusion, our results suggest that “low stress handling” decreases frequency of behaviours related to fear and could improve welfare related to human interactions in adult dogs. references beerda, b., schilderm m.b.h., van hooff, j.a.r.a.m., de vries, h.w., mol, j.a. 1998. behavioural, saliva cortisol and heart rate responses to different types of stimuli in dogs. applied animal behavior science 58: 365–381. csoltova, e., martineau, m., boissy, a., gilbert, c. 2017. behavioral and physiological reactions in dogs to a veterinary examination: owner-dog interactions improve canine well being. physiology and behavior 177: 270-281. döring, d., roscher, a., scheipl, f., küchenhoff, h., erhard, m.h. 2009. fear-related behaviour of dogs in veterinary practice. veterinary journal 182: 38–43. hekman, j.p., zaras, a.z., dreschel, n.a. 2012. salivary cortisol concentrations and behavior in a population of healthy dogs hospitalized for elective procedures. applied animal behavior science 141: 149-157. lloyd, j.k.f. 2017. minimising stress for patients in the veterinary hospital: why it is important and what can be done about it. veterinary sciences 4(2) doi:10.3390/vetsci4020022. overall, k.l. 2013. fear factor: is routine veterinary care contributing to lifelong patient anxiety? magazine september 1. rexroat, j., benish, k., fraden, j. 1999. clinical accuracy of vet-temptm instant ear thermometer: comparative study with dogs and cats. advances monitor corporation1–4. rizzo, m., arfuso, f., alberghina, d., giudice, e., gianesella, m., piccione, g. 2017. monitoring changes in body surface temperature associated with treadmill exercise in dogs by use of infrared methodology. journal of termal biology 69:64-68. yin, s. low stress handling, restraint and behavior modification of dogs and cats: techniques for patients who love their visits; cattledog publishing: davis, ca, usa, 2009. page 22 www pet behaviour science org creative commons license 4.0 – non commercial – share alike – attribution 2017 | vol.3 | 20 22 evaluation of the syndromic characterization and diagnostic criteria of the concept of anxiety in animal ethology professionals. 2022 vol. 12 1 10 evaluation of the syndromic characterization and diagnostic criteria of the concept of anxiety in animal ethology professionals aline ixtab morales-estrada1*, andrés ducoingwatty2, itzcóatl maldonado-reséndiz3 abstract: there is a wide range of reported signs for anxious patients in veterinary medicine but specific diagnostic criteria to characterize them is lacking. the objective of this study was to determine the variability in the concepts of anxiety and the criteria used to diagnose anxiety disorders in veterinary medicine. a questionnaire was developed to obtain information through direct responses from 31 professionals in the exercise of clinical ethology with questions based on the subject of canine anxiety (definition, manifestations, and diagnosis). an analysis was performed using contingency tables. seventy‑three percent (73%, n=22) of respondents agreed on one definition. the signs most frequently considered in the diagnosis of this disorder were increased vigilance (90.3%, n=28), increased motor activity (87%, n=27), panting, and altered heart rate (bradycardia/tachycardia) (77.4%, n=24). from these results, homogeneity is observed in the concept of anxiety, but with evidence of diagnostic heterogeneity, which can be related to the wide repertoire of signs that are considered in the questionnaire and are present in dogs, as well as the lack of diagnostic criteria and/or tests that can objectively evaluate each patient in order to obtain more uniform and reliable results. a.i. morales‑estrada*,1 a. ducoing‑wa y2 i. maldonado‑reséndiz3 1 clinical ethology residency specialty program, facultad de medicina veterinaria y zootecnia, avenida universidad 3000, colonia universidad nacional autónoma de méxico cu, alcadía coyoacán, 04510, ciudad de méxico, méxico 2 ruminant animal production department, facultad de medicina veterinaria y zootecnia, universidad nacional autónoma de méxico 3 ethology, wildlife and laboratory animal department, facultad de medicina veterinaria y zootecnia, universidad nacional autónoma de méxico. *e‑mail: ixtab1609@gmail.com highlights creative common license 4.0 – non commercial – share alike – attribution page 1 keywords: canine anxiety; dog behavior; animal behavior, behavioral medicine, animal welfare, veterinary behavior pet behaviour science 2022, vol. 12, 1 10 doi:10.21071/pbs.vi12.12928 creative common license 4.0 – non commercial – share alike – attribution www.petbehaviourscience.org • currently there are no specific diagnostic standards found in literature that can help characterize and diagnose dogs with anxiety disorders • most behavioral professionals selected clinical signs that are usually more evident in the patients and especially those that can be observed during the consultation • this study found homogeneity in the concept of anxiety, but great variability in the components of the anxiety disorder that are considered for diagnosis morales-estrada, ducoing-watty, & maldonado-reséndiz pet behaviour science creative common license 4.0 – non commercial – share alike – attribution aline ixtab morales‑estrada andrés ducoing‑wa y itzcóatl maldonado‑reséndiz pet behaviour science 2022, vol. 12, 1 10 doi:10.21071/pbs.vi12.12928 introduction anxiety is a basic emotion arising since infancy which helps us adapt to a probable danger, meaning it is not specifically pathological (beesdo‑baum & knappe, 2012). psychologists and psychiatrists define anxiety as an emotion of threat or apprehension of what may happen in the future, which the individual perceives as uncontrollable and unpredictable, with a long‑lasting response (antony & swinson, 2014; sah, 2017). in contrast, barlow (barlow, 2000) defines it as a state of impotence due to a perceived inability to predict, control, or obtain the desired results in certain outstanding situations or contexts in the future. this emotion can be maladaptive and pathological when presented persistently or for long periods of time, associated with mental anguish or deterioration (american psychiatric association & american psychiatric association, 2000). in humans, there are a variety of anxiety disorders, including generalized anxiety disorder, panic disorder, post‑traumatic stress disorder, and social affective disorder (sah, 2017). in psychiatry, the diagnostic and statistical manual of mental disorders (dsm‑ v) defines anxiety as the presence of two or more of the following symptoms: feeling nervousness or tension, unusual restlessness, difficulty concentrating due to worry, fear of terrible things happening, and individuals feeling losing control of themselves (american psychiatric association, 2014). to avoid over‑ diagnosing anxiety disorders in pediatric patients, the criteria was widened to include a minimum presentation time of anxiety symptoms, stating these must be present for at least six months (tayeh et al., 2016). in veterinary medicine, landsberg et al (landsberg et al., 2013), ogata (ogata, 2016), and overall (overall, 2013) define anxiety as an emotion of apprehension in the face of an anticipated danger or threat. signs are recognized through physiological responses (e.g., autonomic activation, increased heart and respiratory rates, tremors, salivation, gastrointestinal alterations), and behavioral responses (e.g., paralysis, lip licking, yawning, wandering, vocalization, hypervigilance, and restlessness); however, the anxiety‑triggering stimulus is not o en identified. in order to diagnose anxiety disorders, overall (overall, 2013) mentions that both neurophysiological (tachycardia, blood pressure alterations, mydriasis, vasodilatation, salivation, muscular tension, etc.) and behavioral (increased vigilance, exploration, motor activity, aggression, etc.) markers must be considered. there is a wide range of signs reported in patients suffering from anxiety disorders, yet no specific diagnostic criteria to characterize patients with this disorder were found at the time of this research. therefore, it is likely that physicians dedicated to the area of clinical ethology have a heterogeneous concept of anxiety, its manifestations, and its diagnostic criteria. the objective of this study was to determine the variability in the concepts of anxiety and the criteria used to diagnose anxiety disorder in dogs. page 2 material and methods participants a questionnaire was sent electronically to 130 veterinarians, researchers in animal psychology/behavior or animal science, and biologists dedicated to the area of clinical ethology. this larger sample size was considered because a significant non‑responsiveness was expected given the way the questionnaire was applied. approximately between one third and one fourth of participants were expected to reply. any number over 30 respondents would ensure the possibility of making inferences about the variables studied. the participantsʹ data were obtained through webpages of associations and institutions of veterinary ethology professionals worldwide: animal behavior society (abs) (www.animalbehaviorsociety.org), european board of veterinary specialization (ebvs) (h ps://ebvs.eu/), american college of veterinary behaviorists (acvb) (www.dacvb.org/), school of veterinary medicine and animal zootechnics ‑ department of ethology, wildlife and laboratory animals unam (h p://www.fmvz.unam.mx) and, linkedin (linkedin.com). this last webpage was considered since it displays users’ information and work experience, a er performing a search for veterinarians practicing clinical ethology. questionnaire the instrument used to obtain information was specifically designed for use in the present study under the google® forms platform. this questionnaire compiled conceptual information and criteria used by clinical ethologists to diagnose anxiety disorders in dogs. it contained questions related to the definition of anxiety and its diagnostic characterization. the questions were answered using a combination of checkboxes, multiple choice, and short answers. the first part of the questionnaire focused on the participants’ general information (gender, country, profession, current occupation, place of professional practice, time working in clinical ethology) and the number of cases they a end per year. the second part was based on the subject of anxiety (definition, manifestations, and diagnosis). the questionnaire is comprised of 13 multiple‑choice and checkbox questions, four of which included the option to choose “other” and input the response in the respondents’ own words, and one open question to include techniques used to characterize anxiety. the questionnaire was sent 130 times and was created based off the information currently available regarding anxiety in dogs. a qualitative validation was performed, defining the concepts to be measured and refining the questions through expert judges, but evaluating the internal consistency of the instrument was impossible. statistical analysis the information obtained in the present study was evaluated using means of analysis for contingency tables and obtaining association measures for the creative common license 4.0 – non commercial – share alike – attribution page 3 2022 vol. 12 1 10 www.petbehaviourscience.org aline ixtab morales‑estrada andrés ducoing‑wa y itzcóatl maldonado‑reséndiz pet behaviour science 2022, vol. 12, 1 10 doi:10.21071/pbs.vi12.12928 variables of years of experience and number of cases a ended against each one of the signs considered by the respondents (seoane et al., 2007). results thirty‑two responses were obtained, although one of them was discarded because 80% of the questionnaire had not been answered. of the 31 remaining questionnaires, 65% (n = 20) were answered by women and 35% (n = 11) by men. according to the country of origin, 26% (n = 8) were from the united states of america, 19% (n = 6) from mexico, 13% (n = 4) from spain, and another 13% from italy; other countries represented were belgium, argentina, brazil, chile, france, and the netherlands. most of the participants were veterinarians (91%, n = 28), the rest were biologists, researchers, and animal science professionals. according to their experience, most of the participants had been working in ethology between 11 and 15 years (23%, n = 7), between 6 to 10 years (22%, n = 7), between 16 and 20 years (16%, n = 5), and more than 30 years (16%, n = 5) (figure 1). creative common license 4.0 – non commercial – share alike – attribution page 4 figure 1. participantsʹ work experience in the area of clinical ethology the definition of anxiety included in the questionnaire that the highest percentage of respondents (73.3%, n = 22, p < 0.01) agreed with was ʺit is an emotion of threat or apprehension of what may happen in the future, which the individual perceives as uncontrollable, unpredictable, and with a lasting responseʺ (figure 2). regarding the signs that characterize an anxiety disorder, 29% (n = 9) associated 16 to 20 signs, 26% (n = 8) associated 11 to 15 signs, and 22% (n = 7) selected more than 20 signs. based on the participants’ responses, the signs most associated with an anxious pet behaviour science morales-estrada, ducoing-watty, & maldonado-reséndiz aline ixtab morales‑estrada andrés ducoing‑wa y itzcóatl maldonado‑reséndiz pet behaviour science 2022, vol. 12, 1 10 doi:10.21071/pbs.vi12.12928 state were increased vigilance (90.3%, n = 28), increased motor activity (87%, n = 27), panting, and altered heart rate (bradycardia/tachycardia) with 77.4% (n = 24) each (figure 3). creative common license 4.0 – non commercial – share alike – attribution page 5 figure 2. most accurate definitions of anxiety in the area of clinical ethology as selected by the respondents figure 3. associated anxiety signs that were mentioned by the participants 2022 vol. 12 1 10 www.petbehaviourscience.org aline ixtab morales‑estrada andrés ducoing‑wa y itzcóatl maldonado‑reséndiz pet behaviour science 2022, vol. 12, 1 10 doi:10.21071/pbs.vi12.12928 all questions but one was multiple‑choice, including the signs seen in figure 3. since some concepts may seem similar, the definitions are provided. for instance, increased vigilance refers to the patient evaluating their environment in order to perceive any threats (robine e & ha, 2001), which can be expressed as a heightened response if the patient is touched or interrupted. on the other hand, increased scanning means the patient moves their eyes or head to constantly scan any activities going around in their environment (hammerle et al., 2015). based on the question related to the number of signs, a patient must present in order to be suffering from an anxiety disorder, 47% (n = 14) answered that patients must present between 1 and 5 signs, while 40% (n = 12) replied that the number of signs is not relevant. when asked what their diagnostic criteria were, 80% (n = 24) responded that they did not use a test to characterize anxiety disorders, and only 20% (n = 6) responded that they did use tests. some tests included for diagnosing anxiety disorders were the exit test for separation anxiety, ignoring the patient for 30 minutes during the consultation, the eded scale (evaluation of a dogʹs emotional disorder), validated questionnaires (such as noise fears/phobias), and a brief exposure to potentially threatening stimuli (always in the presence of the owner). lastly, two significant association were found: one between the participants’ experience in the area of clinical ethology (6 to 10 years, 11 to 15 years) and two signs: increased exploration (p = 0.008), and shuddering (p = 0.018). the other: between the number of patients per year (1‑200) and 2 signs: increased exploration (p = 0.05) and urination (p = 0.08). discussion the definition most selected by the respondents was ʺ(anxiety) is an emotion of threat or apprehension of what may happen in the future, which the individual perceives as uncontrollable, unpredictable, and with a lasting response.ʺ this definition was built from three other similar definitions proposed by three veterinary behaviorists (landsberg et al., 2013; ogata, 2016; overall, 2013). therefore, this can be associated to a term mostly known in the area of ethology with more frequent uses within the professional practice. based on the results, the hypothesis that there would be heterogeneity in selecting a definition for anxiety is discarded since most of the participants were inclined towards the same definition, although these results are not conclusive, and it cannot be said that there is a consensus. regarding the question related to the signs that characterize an anxiety disorder, most of the respondents considered between 16 and 20 signs, followed by 11 to 15 signs, and more than 20 signs ranked third place in their selection. since these are high numbers, it is clear that there are many manifestations considered by the participants, leading to heterogeneity in establishing the probable diagnosis of a patient suffering from an anxiety disorder. creative common license 4.0 – non commercial – share alike – attribution page 6 pet behaviour science morales-estrada, ducoing-watty, & maldonado-reséndiz aline ixtab morales‑estrada andrés ducoing‑wa y itzcóatl maldonado‑reséndiz pet behaviour science 2022, vol. 12, 1 10 doi:10.21071/pbs.vi12.12928 mendoza (mendoza, 2011) characterized behavioral alterations in 125 dogs diagnosed with generalized anxiety disorder (gad), finding muscle tension (76.80%), aggressiveness (66.40%), gastrointestinal problems (vomiting, diarrhea, chronic gastritis, etc.) (36.80%), hyperactivity (wandering, problems staying still, inability to relax) (32.80%), hyper‑vigilance (20.80%), and with sleep problems (non‑ continuous sleep, wakes up easily, has no naps during the day) (16.80%). in the present study, increased vigilance, increased motor activity, wheezing, altered heart rate (bradycardia/tachycardia), lip licking, vocalization, increased monitoring of other individualsʹ actions, yawning, and sleep disturbance were selected more frequently by participants, so it can be observed that the participants selected signs that are usually more evident in the patients and especially observed during the consultation. it should be noted that mendoza studied generalized anxiety disorder so it would be expected that clinicians had selected the signs with the same percentage of their presentation. on the other hand, storengen and collaborators (storengen et al., 2014) conducted a study with dogs diagnosed with separation anxiety disorder (n=215) in which their owners reported the signs they observed most frequently. these signs were: vocalization (83.2%, n=163), destruction (36.4%, n=71) and excessive motor activity (26.2%, n=51). destruction, despite existing in an anxiety disorder, was not mentioned by the questionnaire respondents, which could be associated to the fact that this sign usually appears only in patients with separation anxiety and no other types of anxiety disorders. however, both vocalization and excessive motor activity were also selected more frequently in the present study, which could be related to these two signs being more easily observed by the clinician. based on the number of signs a patient must present to be suffering from an anxiety disorder, most responded that they use between 1 and 5 signs, a smaller range. mendoza (mendoza, 2011) focused only on 6 signs proposed by the diagnostic and statistical manual of mental disorders (dsm) and the international classification of diseases and found that 58% of patients presented 0 to 2 signs, and 37% presented between 3 and 5. an important point to highlight from this article is the question of how a patient with generalized anxiety disorder could be diagnosed with 0 to 2 signs, when the dsm mentions that adult patients must present three or more out of six signs of anxiety to be diagnosed, and at least one sign for the diagnosis of children. furthermore, if the patient presented no signs, it would mean that the patient could be diagnosed with gad when there is no evidence of any signs pointing to this disorder. most respondents answered that they do not use complementary diagnostic tests to characterize anxiety; this result can be related to the fact that several of the respondents were from the same country, and they could have received training or education from the same source. all this could be associated with the diagnostic differences that may exist between graduates from the american college of veterinary behavior (dacvb) and the european college of animal welfare and behavioral medicine (ecawbm) (martin et al., 2014), but there was no significant association between these variables. creative common license 4.0 – non commercial – share alike – attribution page 7 2022 vol. 12 1 10 www.petbehaviourscience.org aline ixtab morales‑estrada andrés ducoing‑wa y itzcóatl maldonado‑reséndiz pet behaviour science 2022, vol. 12, 1 10 doi:10.21071/pbs.vi12.12928 only 20% (n=6) of the respondents mentioned that they use tests to evaluate a patient as suffering from an anxiety disorder, such as the exit test for separation anxiety; ignoring the patient for 30 minutes during consultation; the eded (evaluation of a dogʹs emotional disorder) scale; validated questionnaires (noise fear/phobia); and brief exposure to potentially threatening stimuli always in the presence of the owner. unfortunately, such tests have not been evaluated and validated for objective use in the area of clinical ethology. even though both the research and practice of veterinary behavioral medicine has grown significatively in the past 20 years, this field still lacks standardized protocols for the adequate diagnosis of different behavioral problems frequently reported by pet owners (de assis et al., 2020). due to the aforementioned, and with patients showing a significant variability in the number and types of signs presented, there is no clarity regarding specific signs or groups of signs that may be commonly found in patients with anxiety disorder. results obtained in this study suggest some heterogenicity when characterizing a patient suffering from an anxiety disorder. this could be related to the wide array of signs selected in the questionnaire that can be present in dogs presenting anxiety. this heterogenicity highlights the lack of tests and/or criteria that could be used to objectively evaluate each patient, thus obtaining more uniform and reliable results. conclusions this study found homogeneity regarding the definition of anxiety, which could be a useful start for clinical ethologists to define a patient as suffering from an anxiety disorder and have a basis to make a diagnosis. some signs were selected more frequently by clinicians: increased vigilance, increased motor activity, and panting. despite the consensus in the definition of anxiety and some of its signs, other results obtained in this study suggest a heterogeneity in the way of characterizing a patient with anxiety disorder. there are no standardized tests or parameters for the diagnosis of anxiety disorders in dogs, so further research is needed to develop specific criteria to diagnose patients objectively. it is important to conduct subsequent studies in order to characterize an anxiety disorder in patients among all clinical ethologists, to investigate why tests are not currently being used to help guide or confirm a diagnosis of anxiety disorder in dogs, and to build a universal definition and signalment for anxiety in the area of animal behavior. given the sample size in this study, the results must be interpreted with caution. creative common license 4.0 – non commercial – share alike – attribution page 8 pet behaviour science morales-estrada, ducoing-watty, & maldonado-reséndiz aline ixtab morales‑estrada andrés ducoing‑wa y itzcóatl maldonado‑reséndiz pet behaviour science 2022, vol. 12, 1 10 doi:10.21071/pbs.vi12.12928 acknowledgments to the national council of science and technology (consejo nacional de ciencia y tecnología, conacyt) for the scholarship they granted to aline morales during the specialty, and to the universidad nacional autónoma de méxico. also, to flor ortiz and mish castillo for their valuable observations. funding funding for this project was provided by the authors references american psychiatric association (ed.). (2014). guía de consulta de los criterios diagnósticos del dsm‑5. american psychiatric publishing. american psychiatric association, & american psychiatric association (eds.). (2000). diagnostic and statistical manual of mental disorders: dsm‑iv‑tr (4th ed., text revision). american psychiatric association. antony, m. m., & swinson, r. p. (2014). manual práctico para el tratamiento de la timidez y la ansiedad social: técnicas demostradas para la superación gradual del miedo. desclée de brouwer. barlow, d. h. (2000). unraveling the mysteries of anxiety and its disorders from the perspective of emotion theory. american psychologist, 55(11), 1247‑1263. h ps://doi.org/10.1037/0003‑066x.55.11.1247 beesdo‑baum, k., & knappe, s. (2012). developmental epidemiology of anxiety disorders. child and adolescent psychiatric clinics of north america, 21(3), 457‑478. h ps://doi.org/10.1016/j.chc.2012.05.001 de assis, l. s., matos, r., pike, t. w., burman, o. h. p., & mills, d. s. (2020). developing diagnostic frameworks in veterinary behavioral medicine: disambiguating separation related problems in dogs. frontiers in veterinary science, 6, 499. h ps://doi.org/10.3389/fvets.2019.00499 hammerle, m., horst, c., levine, e., overall, k., radosta, l., ra er‑ritchie, m., & yin, s. (2015). 2015 aaha canine and feline behavior management guidelines*. journal of the american animal hospital association, 51(4), 205‑221. h ps://doi.org/10.5326/jaaha‑ms‑6527 landsberg, g. m., hunthausen, w. l., & ackerman, l. j. (2013). behavior problems of the dog and cat (third edition). saunders/elsvier. martin, k. m., martin, d., & shaw, j. k. (2014). small animal behavioral triage: a guide for practitioners. veterinary clinics of north america: small animal practice, 44(3), 379‑399. h ps://doi.org/10.1016/j.cvsm.2014.01.004 creative common license 4.0 – non commercial – share alike – attribution page 9 2022 vol. 12 1 10 www.petbehaviourscience.org aline ixtab morales‑estrada andrés ducoing‑wa y itzcóatl maldonado‑reséndiz pet behaviour science 2022, vol. 12, 1 10 doi:10.21071/pbs.vi12.12928 mendoza, b. (2011). diagnostico del trastorno de ansiedad generalizada (tag) en perros teniendo como base alteraciones en los signos fisiológicos y conductuales [universidad nacional autónoma de méxico]. h p://docplayer.es/17658471‑ universidad‑nacional‑autonoma‑de‑mexico.html ogata, n. (2016). separation anxiety in dogs: what progress has been made in our understanding of the most common behavioral problems in dogs? journal of veterinary behavior, 16, 28‑35. h ps://doi.org/10.1016/j.jveb.2016.02.005 overall, k. l. (2013). manual of clinical behavioral medicine for dogs and cats. elsevier. robine e, r. l., & ha, j. c. (2001). social and ecological factors influencing vigilance by northwestern crows, corvus caurinus. animal behaviour, 62(3), 447‑ 452. h ps://doi.org/10.1006/anbe.2001.1772 sah, p. (2017). fear, anxiety, and the amygdala. neuron, 96(1), 1‑2. h ps:// doi.org/10.1016/j.neuron.2017.09.013 seoane, t., martín, j. l. r., martín‑sánchez, e., lurueña‑segovia, s., & alonso moreno, f. j. (2007). capítulo 7: estadística: estadística descriptiva y estadística inferencial. semergen ‑ medicina de familia, 33(9), 466‑471. h ps://doi.org/ 10.1016/s1138‑3593(07)73945‑x storengen, l. m., boge, s. c. k., strøm, s. j., løberg, g., & lingaas, f. (2014). a descriptive study of 215 dogs diagnosed with separation anxiety. applied animal behaviour science, 159, 82‑89. h ps://doi.org/10.1016/j.applanim.2014.07.006 tayeh, p., agámez, p. m., & chaskel, r. (2016). trastornos de ansiedad en la infancia y la adolescencia. precop scp, 15, 18. creative common license 4.0 – non commercial – share alike – attribution page 10 pet behaviour science morales-estrada, ducoing-watty, & maldonado-reséndiz aline ixtab morales‑estrada andrés ducoing‑wa y itzcóatl maldonado‑reséndiz pet behaviour science 2022, vol. 12, 1 10 doi:10.21071/pbs.vi12.12928 influence of dog presence on the tolerance and evaluation of aversive stimulation an evaluation of respondent conditioning procedures to decrease barking in an animal shelter payen, s. w*. and assemi, k.s. highlights • many animals relinquished to shelters are relinquished due to problem behavior. • a common problem behavior in dog shelters is barking. • researchers tested the effectiveness of a simple respondent conditioning procedure in order to reduce the noise level of barking in a dog shelter. pet behaviour science | 2017, vol.3, 19 – 24 doi: 10.21071/pbs.v0i3.5758 payne s. w*. and assemi, k.s. california state university, fresno usa paper research * corresponding author: spayne@csufresno.edu keywords: animal shelter, barking, dogs, respondent conditioning • the overall noise levels in the shelter decreased as a result of intervention. • reduction in noise may increase time adopters spend in dog area and increase adoptions. 1. introduction approximately 7.6 million animals are residing in animal shelters in the united states at any given time (american society for the protection and care of animals, 2016). in addition, approximately 25% of animals relinquished to animal shelters are relinquished due to engaging in some form of problem behavior (kwan & bain, 2013; salman et al., 1998). lepper, kass, and hart (2002) found that dogs relinquished to animal shelters due to behavior problems were less likely to be adopted than animals relinquished for non-behavioral reasons. therefore, it may be important to reduce the problem behavior of animals in shelters, which could lead to increased adoption rates. page 19creative common license 4.0 – non commercial – share alike – attribution 2017 | vol. 3 | 19 24 abstract a common problem behavior in animal shelters is excessive noise from barking, which can regularly exceed 100dbs. noise levels in animal shelters are correlated with increased stress in dogs, which may lead to increased problem behavior and a decrease in adoption. the purpose of the current study was to evaluate the use of respondent conditioning procedures to reduce barking noise level in an animal shelter by pairing a door chime with edible items. following a baseline and neutral stimulus phase, the door chime was paired with edible items over a period of three weeks. following this pairing phase, the pairing was stopped to determine if the door chime would act as a conditioned stimulus and reduce barking. these procedures were replicated following an additional baseline phase. overall, the procedure was effective in reducing the noise level of the kennel area as compared to baseline levels. implications and future research areas are discussed. http://www.petbehaviourscience.org/ one relatively common problem behavior in animal shelters is excessive barking. sales et al. (1997) found that the noise level in dog kennels in animal shelters regularly reaches over 100 decibels (dbs), mainly due to excessive barking. hearing damage in humans can occur at noise levels of 85 dbs or greater, which is well below the levels commonly seen at animal shelters (national institute on deafness and other communication disorders, 2016). in addition, excessive noise can cause both psychological and physical stress on subjects in animal shelter. (coppola, enns, & grandin, 2006). due to this increased stress, it is possible that animals may engage in higher levels of problem behavior (dreschel & granger, 2005), which may decrease their chances of adoption. despite excessive barking being a relatively common problem at animal shelters, little research has been conducted to attempt to reduce excessive noise in kennel areas. several studies have shown that playing different genres of music (e.g., classical, reggae, and soft rock) in animal shelters can reduce the amount of barking in the short term (e.g., bowman kogan, schoenfeld-tacher, & simon, 2012; wells, graham, & hepper , 2000). however, bowman et al. (2015) found that habituation to classical music occurred rapidly, and the behavioral effects did not maintain. others have suggested redesigning shelters and shelter policies to reduce problem barking. coppola et al. (2006) suggested redesigning shelter environments, including using soundproofing materials, in order to reduce the overall noise. in addition, hewison et al. (2014) demonstrated that by restricting access to the dog kennel area to one adopter at a time with staff supervision, noise was substantially reduced. however, it should be noted that many animal shelters depend on public funding or donations, and may not have the financial resources to redesign their kennel areas or the staff necessary to restrict access to the kennel area. therefore, solutions that require fewer monetary and staff resources may be more beneficial to animal shelters. protopopova and wynne (2015) conducted a study to increase appropriate behavior that was correlated with adoption in the kennel area of an animal shelter. across two experiments, the authors assessed the effects of a response-dependent reinforcement procedure (differential reinforcement of other behavior [dro]) and a response-independent procedure (noncontingent reinforcement [ncr]). in the dro condition, experimenters would stand at the front of the kennel and wait for the dog to stop engaging in problem behavior (including barking). the experimenter would then deliver an edible item to the dog. in the ncr condition, the experimenter would stand at the front of the kennel and deliver an edible item, regardless of the dog’s behavior. the authors found that both procedures were equally effective in reducing problem behavior. the authors suggested that ncr might be a good method for decreasing problem behavior, because it requires no special training in behavioral observation. the authors also suggested that the effects of the ncr procedure might be due to respondent conditioning, and that the pairing of the edible item with the approaching experimenter may have elicited responding that was incompatible with problem behavior. it is possible that pairing the edible item with another stimulus, such as a door chime, could alleviate the need for an additional person, especially given the potential staffing issues in animal shelters. the purpose of the current study was to extend the results of protopopova and wynne (2015) by explicitly conditioning a previously neutral stimulus (a door chime) with the delivery of edible items (unconditioned stimulus) to determine if the door chime would elicit reduced levels of barking in an animal shelter following the conditioning procedure. 2. methods subjects subjects were 50 dogs (canis lupus familiaris) of various breeds and ages (range: 6 months to 12 years), including mixed breeds. the mean length of stay in the shelter for dogs in the study was 6 months (range: 2 weeks to 5 years). all dogs in the shelter were spayed/neutered. throughout the course of the study, several dogs were adopted and/or introduced to the animal shelter, but the global population remained stable. no steps were taken to control for a stable population throughout the study, as we did not want to prevent the adoption or intake of any shelter dogs. page 20 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2017 | vol.3 | 19 24 additionally, most animal shelters have regular intakes/adoptions, and thus the results of the current study would have more generality to other shelters where this is commonplace. setting all sessions took place in the kennel area of a local nokill animal shelter in fresno, california. the kennel area contained 26 1m x 4m kennels with access to an outdoor kennel area of the same size. dogs could move freely between the outdoor area and indoor area throughout the observation times in the study. the kennels had opaque concrete walls on two sides, and chain-link doors at either end. dogs were housed between one to four dogs per kennel depending on size and temperament, although a majority of the dogs were housed alone. kennels were situated in such a way that dogs could see other dogs in the kennel across from them. kennels contained either raised-platform beds or soft beds. no enrichment items were provided to the dogs per shelter policy. kennels were cleaned twice daily prior to the shelter opening (8:00am-10:00am) and after the shelter closed to the public (6:00pm8:00pm). dogs were fed once per day following the closure of the shelter to the public. the kennel area had three doors, one of which could be entered by the general public and two that were restricted to shelter staff and volunteers. response definition and measurement the dependent variable in the study was the noise level due to barking. barking was measured in decibels (dbs) continuously on a second-by-second basis using a wensn® digital sound level meter (item number ws1361c; manufactured in the u.s.a.) with a range of 30db to 130db. for all sessions, the sound level meter was placed in an unobtrusive location near the center of the kennel area. the sound level meter saved data to a pc based computer program. equipment throughout the study, a wensn® digital sound level meter (described above) was used to record the sound level in the kennel area. additionally, three sabre® digital door chimes (item number hs-dwa2; manufactured in china) were used in the neutral stimulus, pairing procedure, and post-pairing conditions. the door chimes consisted of two parts, a sensor and receiver. the sensor was attached to the door and the receiver was attached to the doorframe on all of the entrances to the kennel area. when the door was opened, the sensor and receiver were separated and the door chime emitted a 120 db tone for 0.5 s. procedures all sessions began when the sound level meter was activated and lasted for 5 minutes. following the activation of the sound level meter, the experimenter left the kennel and did not return until the end of the session. sessions were conducted at varying times per day during the operating hours of the animal shelter when the shelter was open to the public (10:00am4:30pm), with one to three sessions per day, three to five days per week. care was taken to ensure that no single time period was favored in the recording. an abcac (a: basline; b: neutral stimulus; c: postpairing) reversal design was used for experimental control. baseline. during baseline, no programmed stimuli were in place in the kennel area. sound levels were recorded for the duration of the sessions. neutral stimulus (ns). during the neutral stimulus condition, a digital door chime was placed on all three access points into the kennel area such that the door would chime when any door was opened. there was no programmed pairing of this stimulus with any other stimulus under the control of the experimenter. pairing procedure. following the ns condition, the pairing procedure began. during this procedure, the experimenter walked to the front of each kennel. the experimenter activated the digital door chime and provided an edible treat (which varied based on the treats available from donations to the shelter; e.g., milkbone® soft and chewytm, bil-jac® original recipe, pup-peroni® ) to each dog in the kennel. following the delivery of the edible treat, the door chime was sounded two additional times during consumption to provide additional pairings of the chime with the treats. each dog throughout the study accepted all page 21 respondent conditioning to reduce barking creative common license 4.0 – non commercial – share alike – attribution payne and assemi https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science treats. this procedure was conducted once per day, three-to-five days per week for three weeks (for a total of 45 pairings). during the replication of this phase, the procedure was conducted once per day, three-to-five days per week for one week. during this condition, the dogs consumed the treats throughout the time of the two additional chimes. post-pairing (cs). the procedures of the cs condition were identical to the ns condition, with the exception that the pairing procedure was conducted prior to the cs condition. data analysis data were analyzed throughout the course of the study using visual analysis of the graphical depiction of the data. data were also summarized using means, tukey’s tri-mean, and ranges for each phase. statement of approval the procedures described in this manuscript were approved by the institutional animal care and use committee (iacuc) at california state university, fresno. conflict of interests statement no conflicts of interest were present in the conducting of this study. 3. results and discussion the results of the study suggest that the respondent conditioning procedure was effective in decreasing the noise intensity of the kennel area (see figures 1 and 2). during baseline and the ns condition, the noise of the kennel area was variable across sessions, but maintained at relatively high levels (x = 85.1 dbs; tm= 82.6 dbs; range= 55.3 dbs-111.4 dbs), often above the threshold for damage to hearing in humans (85 dbs). following the pairing procedure, the noise level reduced to relatively low levels in the cs condition (x = 68.1 dbs; tm= 67.8 dbs; range= 36.1 dbs-106.1 dbs), with noise levels comparable to normal speech level. we next returned to baseline and sound levels initially remained at relatively low levels, possibly due to carryover from the previous condition (i.e., stimulus control), but gradually increased to levels similar to the initial baseline (x = 71.3 dbs; tm= 70.9 dbs; range = 46.4 dbs 106.3 dbs). finally, we conducted the pairing procedure for one week, and in the subsequent cs condition, we saw the noise levels decrease to levels similar to the initial cs condition (x = 70.1 dbs; tm= 70.8 dbs; range= 48.4 dbs-102.5 dbs). page 22 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2017 | vol.3 | 19 24 figure 1. the mean dbs of noise per session in the kennel area across conditions. the dotted line represents the threshold at which hearing damage in humans can occur following prolonged exposure. overall, the results were similar to those of protopopova and wynne (2015). the non-contingent delivery of edible items paired with the door chime reduced barking behavior in the animal shelter when only the door chime was present. in addition, mean noise levels were reduced below the threshold of hearing damage in humans. the decrease in noise in the kennel area of the animal shelter may reduce physiological stress on the animals and humans in the kennel area. it is also possible that potential adopters may spend more time in the kennel area and be more likely to adopt an animal. there are several possible mechanisms by which barking was reduced. first, following the pairing of the door chime and the edible items, it is possible that the door chime became a conditioned stimulus that elicited responding that was incompatible with barking (such as salivation). second, the door chime, through its pairing with the edible item, may have become a conditioned reinforcer. it is possible that humans entering the kennel area were previously aversive to the dogs, and barking may have been maintained by negative reinforcement in the form of escape from humans. following the pairing of the door chime with the edible item, it is possible that the door chime served as an abolishing operation for the escape-maintained behavior and reduced the aversiveness of humans entering the kennel area and the kennel area in general (similar to enrichment). future researchers should evaluate these specific mechanisms to determine the mechanism by which the pairing reduced barking. there are a few limitations to the current study. first, our data collection mechanism did not discriminate between barking and other sounds in the kennel area. anecdotally, almost all of the sound in the kennel area was from barking, and thus it is unlikely that other extraneous noise influenced the results in a meaningful way. second, dogs were adopted and introduced throughout the course of the study. although not documented, it is possible that dogs were introduced following the pairing procedures, and thus the dogs had never come into contact with the edible item paired with the door chime. third, the housing of other dogs was not controlled. while this may have increased the ecological validity of the current study, it is also possible that the presence of other dogs in the same kennel may have influenced levels of barking during the study. fourth, the respondent conditioning procedure used in this study did not specifically target an unconditioned response, so it was unclear by what mechanism the barking decreased. future researchers should continue to evaluate respondent conditioning procedures to reduce problem behavior and increase appropriate behavior in shelter animals. protopopova and wynne (2015) found that the response-independent delivery of edible items was effective at reducing many different problem behaviors. it is possible that other problem behaviors were reduced in the current study, but due to the nature of measurement we did not obtain this information. therefore, additional measures including observational measures should be used to determine if respondent conditioning procedures have effects on other potentially problematic behaviors in addition to barking in animal shelters. additionally, future researchers should train animal shelter staff and volunteers to use these procedures. it is possible that pairing procedure could be conducted within the normal feeding routines of the shelter and may help maintain the effects of the respondent conditioning procedure. future researchers should also conduct social validity measures to determine if the procedures decrease problem behavior and increased the “perceived adoptability” of the animals in a socially significant way. finally, future researchers should determine the effects of various procedures to reduce problem behavior on the overall adoption rates of animals in animal shelters. page 23 respondent conditioning to reduce barking creative common license 4.0 – non commercial – share alike – attribution payne and assemi figure 2. the mean dbs of noise in the kennel area between the baseline and post-pairing conditions. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science 4. references american society for the protection and care of animals (2016). pet statistics. retrieved from http://www.aspca.org/animal-homelessness/shelterintake-and-surrender/pet-statistics. bowman, a., dowell, f. j., evans, n.p., & scottish spca (2015). ‘four seasons’ in a rescue centre: classical music reduces environmental stress in kenneled dogs. physiology & behavior, 143, 70-82. bowman, a., scottish spca, dowell, f. j., & evans, n. p. (2017). the effect of different genres of music on the stress level of kenneled dogs. physiology and behavior, 171, 207-215. coppola, c. l., enns, r. m., & grandin, t. (2006). noise in the animal shelter environment: building design and the effects of daily noise exposure. journal of applied animal welfare science, 9, 1-7. dreschel, n. a., & grander, d. a. (2005). physiological and behavioral reactivity to stress in thunderstormphobic dogs and their caregivers. applied animal behaviour science, 95, 153-168. hewison, l. f., wright, h. f., zulch, h. e., & ellis, s. l. h. (2014). short term consequences of preventing visitor access to kennels on noise and the behavior and physiology of dogs housed in a rescue shelter. physiology & behavior, 133, 1-7. kogan, l.r., schoenfeld-tacher, r., & simon, a.a. (2012). behavioral effects of auditory stimulation on kenneled dogs. journal of veterinary behavior, 7, 268-275. kwan, j. y., & bain, m. j., (2013). owner attachment and problem behaviors related to relinquishment and training techniques of dogs. journal of applied animal welfare science, 16, 168-183. lepper, m., kass, p. h., & hart, l. a. (2002). prediction of adoption versus euthanasia among dogs and cats in a california animal shelter. journal of applied animal welfare science, 5, 29-42. national institute on deafness and other communication disorders (2016). noise-induced hearing loss. retrieved from https://www.nidcd.nih.gov/health/noise-inducedhearing-loss. protopopova, a., & wynne, c. d. l. (2015). improving in-kennel presentation of shelter dogs through response-dependent and response-independent treat delivery. journal of applied behavior analysis, 48, 590-601. sales, g., hubrecht, r., peyvandi, a., milligan, s., & shield, b. (1997). noise in dog kenneling: is barking a welfare problem for dogs. applied animal behavior science, 52, 321-329. salman, m.d., new, j.c., scarlett, j.m., kass, p.h., hetts, s., ruch-gallie, r. (1998). human and animal factors related to the relinquishment of dogs and cats in 12 selected animal shelters in the usa. journal of applied animal welfare science, 1, 207-226. wells, d. l., graham, l, & hepper, p. g. (2002). the influence of auditory stimulation on the behaviour of dogs housed in a rescue shelter. animal welfare, 11, 385393. page 24 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2017 | vol.3 | 19 24 biomedical scent detection dogs: would they pass as a health technology? biomedical scent detection dogs: would they pass as a health technology? mirkka koivusalo1 and catherine reeve2* pet behaviour science | 2018, vol.6, 1 – 7 doi: 10.21071/pbs.v0i6.10785 mirkka koivusalo and catherine reeve 1 canid and reptile behaviour and olfaction lab. department of psychology & neuroscience, life sciences centre, 1355 oxford street, dalhousie university. halifax, nova scotia. b3h 4r2 canada 2 animal welfare and behaviour school of psychology queen’s university belfast university rd. belfast, northem ireland bt7 1nn review * email: c.reeve@qub.ac.uk keywords: scent detection dogs; canine detection training; biomedical scent detection; health technology assessment; clinical effectiveness; reproducibility. highlights • because biomedical scent detection is a potential tool to identify diseases, in the eyes of regulatory bodies it is comparable to a health technology. • this is the first report evaluating the canines’ sense of smell from the point of view of a health technology. • it is important for researchers to be aware of the required regulatory controls before dogs can have an accredited role in the clinic. • we discuss the current advantages and disadvantages of the method to help guide researchers towards attainable goals. page 1creative commons license 4.0 – non commercial – share alike – attribution 2018 | vol. 6 | 1 7 abstract biomedical scent detection dogs identify the scent profiles of diseases, such as cancer, diabetes or pathogenic micro-organisms. what the field of biomedical scent detection has been lacking, however, is the assessment of the method from the point of view of a health technology. all health technologies undergo a thorough evaluation of safety, clinical effectiveness and costs, as well as ethical, social, organizational and legal evaluations in some cases. passing these regulatory controls is a pre-requisite before a technology is approved for use in decision-making about patient outcomes. biomedical scent detection has a lot of attractive qualities, such as the sensitivity and specificity of the dogs’ noses, safety and relative cost-effectiveness. however, the method also has various challenges, in particular regarding its clinical effectiveness. the most pertinent issues to address before the dogs would pass as a health technology are standardization of the training techniques, both intraand inter-dog reproducibility, and generalization of the detection task to the early stages of disease progression. we recommend setting realistic goals in terms of what the dogs can and cannot do and adopting a collaborative approach between clinicians and animal psychophysicists. http://www.petbehaviourscience.org/ introduction canines’ sense of smell has been utilized successfully for a long time for different purposes, such as detection of explosives, narcotics, and bed bugs, to name just a few. biomedical scent detection (or medical scent detection) is an emerging method that uses the canine olfactory capacity for identifying the scent profiles of diseases, such as different types of cancers (moser and mcculloch 2010; jezierski et al. 2015), diabetes (gadbois and reeve 2014; hardin et al. 2015) or the presence of pathogenic micro-organisms (bomers et al. 2014; bryce et al. 2017; koivusalo et al. 2017; koskinen et al. 2017). it is the combination of the acuity of the sense of smell with the ability to learn by operant conditioning that makes dogs potential biodetectors for different tasks (pirrone and albertini 2017). the ultimate goal in most cases is to involve dogs as a fast method for the diagnosis or screening of human patient disease. research studies illustrate that dogs are capable of distinguishing the volatile organic compounds characteristic of diseases (rudnicka et al. 2014). an important topic to take into consideration when training and testing dogs for this purpose is a comparison with conventional technologies used in screening and diagnosis. if a dog’s indication is to be used in making decisions on a patient’s health and treatment outcomes, canine scent detection would be comparable to a health technology in the eyes of health organizations and regulatory bodies. the purpose of this review it to evaluate canine scent detection as a medical device, to assess whether it would pass a health technology assessment (hta) as it currently stands and what the most pertinent technical challenges are. definition and quality assessment of health technologies the world health organization (who) defines a health technology as (world health organization 2018): “a health technology is the application of organized knowledge and skills in the form of devices, medicines, vaccines, procedures and systems developed to solve a health problem and improve quality of lives.” amongst the different technologies, the dog’s nose could be categorized as a medical device: a highly sensitive, built-in olfactory detection system used to identify diseases. however, before a medical device or any health technology is granted permission for official clinical use in any country, an approval is required by national health organizations, such as food and drug administration (fda) in the us. the requirements for approval include passing several regulatory controls (us food and drug administration, 2018) that set a high standard for all health technologies. health technology assessment (hta) is an evaluation tool a practical tool for evaluating the quality of health technologies is a process known as health technology assessment (hta), which uses scientific research to inform decision-makers on the introduction and use of a health technology. each country’s regulatory bodies implement hta in their own way, but international organizations have set guidelines for the process (eunethta core model® 2016; health technology assessment international 2018; international network of agencies for health technology assessment 2018). the aspects to be evaluated depend on the technology in question and the assessing regulatory body. the following information, which is pertinent also to biomedical scent detection, is always asked: • description of the health problem and currently available technology for solving it. • technical description of the new technology in detail. • clinical effectiveness: does the new technology work equally well or better than a standard method currently in use? the requirement before granting a permission for marketing is to demonstrate that the technology is at least “substantially equivalent” (fda) to a current standard technology. page 2 www pet behaviour science org creative commons license 4.0 – non commercial – share alike – attribution 2018 | vol.6 | 1 7 • safety: the technology has to be safe not only for the patients but also for the people operating it. potential risks and harms are assessed that will determine the risk classification of the technology. • economic evaluation: value-for-money assessment is conducted to evaluate the costs and benefits of the technology in relation to health-related outcomes as well as the economic burden on the healthcare system and society. • ethical, social, organizational and legal aspects may also be evaluated. how does biomedical scent detection rank in evaluation of the most fundamental criteria? clinical effectiveness if we ask whether dogs can smell cancer or a pathogen, the answer would be a ‘yes’, as dogs can detect diseases with very high levels of sensitivity and specificity (jezierski et al. 2015). but to assess the clinical effectiveness of the technology the question is: can dogs do it with the same reproducibility and accuracy as an analytical instrument already in use, and can patient treatment outcomes be planned on the basis of the results? to be approved by health organizations, several aspects need to be addressed before the answer is ‘yes’ for canine scent detection (fig. 1). here are some of the main challenges that scent detection researchers are trying to solve: 1) standardization of the training and testing methods currently there is no standardized way to train and test the dogs, which leads to variability in results (as discussed by elliker et al. 2014; jezierski et al. 2015; gadbois & reeve, 2016). traditionally dogs have been trained and tested using an alternative forced choice (afc) method, where a line-up or a scent wheel contains one target scent (s+, here, a “disease odour”) and several non-target or distractor scents (s–, a healthy control odour), and then running double-blind trials, where neither the dog nor the dog handler knows the sample positioning and the dog is required to locate and indicate the target, diseased, odour. this method is appropriate for a proof-of-principle study, but in a real screening situation the number of s+ and s– samples is unknown. if the dog has been trained to always find one ‘win’ in a line-up, what happens in a screening situation? studies have shown that when dogs were trained using the afc task of one s+ and four s– samples, and then tested with randomized numbers of s+ and s– samples, page 3 detection dogs in healthcare creative commons license 4.0 – non commercial – share alike – attribution koivusalo & reeve figure 1. evaluation of biomedical scent detection as a health technology https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science the sensitivity and specificity of their performance deteriorated significantly (amundsen et al. 2014; hackner et al. 2016). this illustrates the importance of training and testing the dogs with randomized samples as early in the process as possible. the afc method also has a disadvantage due to a memory load it may impose on the detection dog. gadbois and reeve (gadbois and reeve 2014; 2016) have shown that the accuracy of canines decreases significantly the further down the target is in the lineup because of a tax on the dogs’ memory processing. the goal of biomedical scent detection is to assess the sensory-perceptual abilities of dogs to distinguish scents extremely similar to each other, and not make the process a memory load task of a past event. therefore, to avoid the tax on memory, the use of discrimination tasks of 2 or 3 choices is more favourable (gadbois and reeve 2014; 2016; koivusalo et al. 2017). an even more elegant solution is a pure detection task, i.e. a yes/no, go/no-go system, where the dog is presented with one stimulus at a time, which requires a ‘yes’ or ‘no’ answer (gadbois and reeve 2014, 2016). in this approach the dog always gets a win and the task is a pure sensory task, which increases accuracy. 2) reproducibility an analytical laboratory instrument analyzes hundreds of samples daily with high accuracy and throughput, albeit a certain degree of variability (as discussed by moser & mcculloch 2010). dogs cannot be considered as instruments, and they will never obtain an accuracy of 100% due to their own inherent variability (hackner and pleil 2017). furthermore, the published medical scent detection studies are different from real screening situations, where larger sets of samples would be processed daily, and the reproducibility will be harder to accomplish. this stems from several factors. the work of an ‘analytical’ dog is highly repetitive, and it can be difficult to keep up their motivation, as dogs are subject to boredom, fatigue and external distractions. the job of these dogs is different from the scent detection work in the field (e.g. drugs, explosives, search and rescue), where the dogs get to use more of their natural ‘hunting’ behaviors and the task remains more of a game to them. in contrast to laboratory devices, dogs may try and change their strategies in order to be rewarded at minimum effort leading to inaccuracies during doubleblind trials. the handler’s positive and negative sensations are easily transmitted to the dogs. also, in contrast to analytical devices that detect compounds very specifically, each individual dog can perceive complex scent patterns differently leading to variability between different dogs. finding dogs with an ability to generalize the complex detection task is important. 3) detection of diseases in true unknown samples most published studies have been performed with samples from already diagnosed patients. however, it is crucial to improve the overall survival: earlier cancer detection may permit earlier intervention and detection of colonization by a pathogen prior to an actual infection may enable early treatment and containment as well. therefore, the next step after standardization of training/testing methods is to obtain results with completely unknown samples. it is likely that the scent profile changes between samples from patients with an advanced vs. an earlier stage of disease. amundsen et al. (amundsen et al. 2014) showed that the dogs’ performance is affected when they are tested with patients suspected to have different stages and forms of lung cancer. this finding highlights the importance of training the dogs not only with samples from patients with an advanced or malignant disease but with samples from benign stages of disorders. to teach the dogs the subtle discrimination among different stages of a disease may not be a trivial task, as what truly is a positive sample and what is a negative one can turn out to be a grey area. this is where drawing realistic cut-offs for what dogs can and cannot do should be considered. page 4 www pet behaviour science org creative commons license 4.0 – non commercial – share alike – attribution 2018 | vol.6 | 1 7 4) sample presentation one experimental factor to consider and standardize is the way the biological samples are presented and preserved (hackner and pleil 2017; reeve et al. 2017). also the protocol used (or not used) to clean and handle the scent containers between trials differs widely, but ultimately the goal is that the dogs discriminate the target scent amongst any background scent. what is important to acknowledge in biomedical scent detection is that the dog is constantly learning to discriminate subtle differences between samples, and without due diligence they may accidentally learn to indicate something superfluous or superstitious. to circumvent this, many researchers settle for a set cleaning protocol or using disposable scent containers only (jezierski et al. 2015; koivusalo et al. 2017; koskinen et al. 2017). safety this is one of most attractive qualities of biomedical scent detection: it is very non-invasive in nature, and therefore safe for the patients. for some patients a physical interaction with animals may be a concern, but if the detection work takes place in a laboratory, there is no need for contact between the patients and the dogs. an important risk assessment to be made is the evaluation of the consequences of false positive (dog indicates an s– sample) and false negative (dog does not indicate s+) results. false negative results would impose a more serious safety concern, as it would involve missing patients at risk. cost evaluation another appealing quality of biomedical scent detection is its potential to be more cost-effective than many expensive laboratory methods. for example, arnaud (2016) determined that biopsy-based methods of non-small cell lung cancer biomarker testing can cost upwards of $2,500. the analysis of volatile organic compounds (vocs, amann et al. 2014) in exhaled breath samples is a much less expensive, and less invasive procedure, and has proven a promising avenue for cancer diagnosis (shirasu and touhara, 2011). the analytical techniques used to identify disease-specific vocs, such as spme and gc-ms, however, can be expensive and extensive training is required to perform the analysis and analyze the results (shirasu and touhara, 2011). alternatively, the identification of diseased breath samples by detection dogs requires minimal equipment and the analysis (interpretation of the dogs’ indication behaviour), is straight forward (jezierski et al. 2015). mcculloch et al. (2006) examined canine detection of non-small cell lung cancer in donated breath samples and showed that the dogs were 99% sensitive and specific, providing evidence for a highly promising yet cheaper detection system. also, dogs would be a rational solution in areas where high-tech instrumentbased analysis is unavailable (rooney et al. 2013; jezierski et al. 2015) the disadvantage is that the process of training a dog is a time-consuming and hence a money-consuming process because it can take up to a year to train a dog; fully trained medical alert dogs can cost anywhere between $8000 $20,000 usd (beyondtype1.org). even then, it does not mean that every individual dog is suitable for the task, and even at later stages of testing, some dogs may not show the desired sensitivity and specificity (jezierski et al. 2015). conclusion to be approved as a clinically valid health technology by regulatory bodies, biomedical scent detection has not reached the required standard yet. whether it will, depends on the crucial research that needs to be done. in the current age of high-tech medical devices competing in the market, setting a realistic target for the canines might speed up the process as it is a race of who has the most clinically effective and cost-effective method first. in order to make biomedical scent detection a reliable health technology, understanding of the canine olfactory learning processes is needed both through a clinical research and psychophysical approach. acknowledgements we thank dr. hanni uusi-kerttula for critically reading the manuscript. page 5 detection dogs in healthcare creative commons license 4.0 – non commercial – share alike – attribution koivusalo & reeve https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science this research was supported by an nserc pgs-d to catherine reeve. references amann, a., de lacy costello, b., miekisch, w., schubert, j., buszewski, b., pleil, j., ratcliffe, n., and risby, t. 2014. the human volatilome: volatile organic compounds (vocs) in exhaled breath, skin emanations, urine, feces, and saliva. journal of breath research, 8, 034001. amundsen, t., sundstrom, s., buvik, t., gederaas, o. a. and haaverstad, r. 2014. can dogs smell lung cancer? first study using exhaled breath and urine screening in unselected patients with suspected lung cancer. acta oncologica 53: 307–315. arnaud, a. 2016. costs and outcomes comparison of tissue and blood-based biopsies for the purpose of biomarker testing [abstract]. value in health, 19, a143144. beyond type 1. 2018, july. diabetic alert dogs. retrieved from: https://beyondtype1.org/diabetic-alertdogs-dads/ bomers, m. k., van agtmael, m. a., luik, h., vandenbroucke-grauls, c. m. j. e. and smulders, y. m. 2014. a detection dog to identify patients with clostridium difficile infection during a hospital outbreak. journal of infection 69: 456–461. bryce, e., zurberg, t., zurberg, m., shajari, s. and roscoe, d. 2017. identifying environmental reservoirs of clostridium difficile with a scent detection dog: preliminary evaluation. journal of hospital infection 97: 140–145. elliker, k.r., sommerville, b.a., broom, d.m., neal, d.e., armstrong, s., & williams, h.c. 2014. key considerations for the experimental training and evaluation of cancer odour detection dogs: lessons learnt from a double-blind controlled trial of prostate cancer detection. bmc urology 14: 22. eunethta® core model version 3.0. 2016. http://www.eunethta.eu/outputs/hta-core-model-30. accessed march 31, 2018. food and drug administration. 2018. medical devices. https://www.fda.gov/medicaldevices/resourcesforyou/ consumers/default.htm. accessed march 31, 2018. gadbois, s. and reeve, c. 2014. canine olfaction: scent, sign, and situation. in domestic dog cognition and behaviour, 3-29, ed. a. horowitz. berlin, heidelberg: springer-verlag. gadbois, s. and reeve, c. 2016.the semiotic canine: scent processing dogs as research assistants in biomedical and environmental research. dog behaviour 2: 26–32. hackner k., errhalt p., mueller m.r., speiser m., marzluf b.a., schulheim a., schenk p., bilek j. and doll t. 2016. canine scent detection for the diagnosis of lung cancer in a screening-like situation. journal of breath research 10: 46003. hackner, k. and pleil, j. 2017. canine olfaction as an alternative to analytical instruments for disease diagnosis: understanding ‘dog personality’ to achieve reproducible results. journal of breath research 11: 1–5. hardin, d. s., anderson, w. & cattet, j. 2015. dogs can be successfully trained to alert to hypoglycemia samples from patients with type 1 diabetes. diabetes therapy 6: 509–517. health technology assessment international (htai). 2018. https://www.htai.org/. accessed march 31, 2018. international network of agencies for health technology assessment (inahta). 2018. http://www.inahta.org/. accessed march 31, 2018. jezierski, t., walczak, m., ligor, t., rudnicka, j. and buszewski, b. 2015. study of the art: canine olfaction used for cancer detection on the basis of breath odour. perspectives and limitations. journal of breath research 9: 27001. koivusalo, m., vermeiren, c., yuen, j., reeve, c., gadbois, s., & katz, k. 2017. canine scent detection as a tool to distinguish methicillin-resistant staphylococcus aureus. journal of hospital infection 96: 93-96. koskinen, a., koskinen, h., bäck, l., saxen, h. and klockars, t. 2017. a detection dog for paediatric page 6 www pet behaviour science org creative commons license 4.0 – non commercial – share alike – attribution 2018 | vol.6 | 1 7 urinary tract infection caused by escherichia coli. infectious diseases 4235: 1–4. mcculloch, m., jezierski, t., broffman, m., hubbard, a., turner, k., janecki, t. 2006. diagnostic accuracy of canine scent detection in earlyand late-stage lung and breast cancer. integrative cancer therapies, 5, 30-39 moser, e. & mcculloch, m. 2010. canine scent detection of human cancers: a review of methods and accuracy. journal of veterinary behavior 5: 145–152. pirrone, f. and albertini m, 2017. olfactory detection of cancer by trained sniffer dogs: a systematic review of the literature. journal of veterinary behaviour 19: 105–117. reeve, c., wentzell p., wielens, b., jones, c., stehouwer, k. and gadbois s. 2017. assessing individual performance and maintaining breath sample integrity in biomedical detection dogs. behaviuoral processes doi:10.1016/j.beproc.2017.08.008. rooney, n.j., morant, s., & guest, c. 2013. investigation into the value of trained glycaemia alert dogs to clients with type 1 diabetes. plos one 8: e69921. rudnicka, j., walczak, m., kowalkowski, t., jezierski, t., & buszewski, b. 2014. determination of volatile organic compound as potential markers of lung cancer by gas chromatography-mass spectrometry versus trained dogs. sensors and actuators 202: 615-621. shirasu, m., and touhara, k. 2011. the scent of disease: volatile organic compounds of the human body related to disease and disorder. the journal of biochemistry, 150, 257-266. world health organization. 2018. what is a health technology? http://www.who.int/health-technologyassessment/about/healthtechnology/en/. accessed march 31, 2018. page 7 detection dogs in healthcare creative commons license 4.0 – non commercial – share alike – attribution koivusalo & reeve https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science impact of therapy dog visits on outpatient nurse welfare and job satisfaction impact of therapy dog visits on outpatient nurse welfare and job satisfaction stephanie d. clark*, jessica m. smidt, and brent a. bauer pet behaviour science | 2018, vol.6, 8 – 15 doi: 10.21071/pbs.v0i6.11172 stephanie d. clark*, jessica m. smidt, and brent a. bauer * department of integrative medicine and health division of general internal medicine mayo clinic 200 first st sw rochester, mn 55905 research paper * email: clark.stephanie1@mayo.edu keywords: burnout; compassion fatigue; therapy dogs; welfare highlights • outpatient nursing units that experience a therapy dog visit, once a week, self-reported an improvement in emotional wellbeing score, including, depression, stress, happiness, energy level, relaxation, calmness, and overall emotional wellbeing.. • no therapy dog visit group, had the least amount of improvement in the nursing units’ visual analog scale was seen, with six parameters, restlessness, anxiety, depression, relaxation, calmness, and overall emotional wellbeing, being self-reported as worse. page 8creative commons license 4.0 – non commercial – share alike – attribution 2018 | vol. 6 | 8 15 abstract interaction with a therapy dog can decrease blood pressure, heart rate, and improve heart rate variability; due to these responses, it suggests that human-animal interaction can alleviate the stress response. this study aims to observe if the effects of therapy dog visits could alleviate nursing burnout and increase work satisfaction in an outpatient setting. in addition, this study will observe at what visit frequency of therapy dog visits nurses benefited from most. this study is a two-part study, which also observed the salivary cortisol concentrations of the therapy dogs post therapy visit interaction. the study design was a controlled before-after study design with five treatments over the course of six months, each treatment (trt) lasting four weeks: trt a, two therapy dog visits a week; trt b, one visit a week; trt c, two visits; trt d, one visit; and trt e, no visits. four out-patient nursing units were selected and asked to complete a demographic survey, the pet attitude scale-modified, and lexington attachment to pet scale. preand post-treatments, participants completed the human services survey, nursing workplace satisfaction questionnaire, nursing work index (revised), and a visual analog scale. trt a was able to significantly increase the feeling of happiness. in addition trt b, a therapy dog visit once a week, was able to significantly reduce self-reported responses of depression and improve emotional wellbeing. consequently, trt e, control/no therapy dog visits, had the least amount of improvement in the nursing units’ visual analog scale. this study supports the hypothesis that therapy dog visits can help alleviate stress, frustration, feeling drained, and the overwhelming sensation that can come from working in the nursing field. http://www.petbehaviourscience.org/ • participated in the study who currently owned a pet (100%), over 80% believe that owning pets contribute to their happiness. introduction nurse and health care provider burnout has consistently been an upward battle. burnout is described as a syndrome characterized by emotional exhaustion, depersonalization, and a decrease in personal accomplishment (vahey et al., 2004). in addition to decreases in productivity and patient satisfaction, burnout can also lead to psychological distress, an increase in reported complaints, and drug and alcohol abuse (vahey et al., 2004). reports state that over 40% of nurses experience burnout, a higher rate than other health care providers is often a result of unrealistic workloads aiken et al., 2011). while nurses play a primary role in the overall care of patients and daily hospital tasks, the nursing field is inadequately staffed and a main limiting factor in high-quality hospital care (aiken et al., 2002). in addition, these nurses indicated that their stress-coping behaviors included exercise, social support, verbal venting, selfdistraction, and self-blame (shimizutani et al., 2008). in addition to self-care behavior, another strategy that can be easily implemented to decrease nurse burnout is animal-assisted therapy visits. the human-animal bond is growing stronger and the demand for therapy dog visits in a hospital setting is increasing. studies have been conducted observing the beneficial relationship animals have on the elderly, children with autism, hospitalized patients, hospice care, and behavioral conditions (nimer and lundahl, 2007). in particular, one study observed the therapeutic effects of dog visits with hospitalized patients with a psychiatric disorder. the researchers observed that a dog visit significantly reduced the level of anxiety experienced by patients (barker and dawson, 1998). additionally, animal assisted therapy visits can improve quality of life, coping ability, and self-efficacy (berget et al., 2008). animal assisted therapy not only improves mental outlook, but physiologically, 20 minutes of petting a dog can increase oxytocin concentrations, as well as positively affect other endocrine responses, such as a decrease in cortisol, epinephrine, and norepinephrine (beetz et al., 2012). interaction with a friendly animal can also decrease blood pressure, heart rate, and improve heart rate variability; due to these responses, it suggests that human animal interaction can alleviate a stress response (beetz et al., 2012). with this supporting data, animal-assisted therapy visits could provide an easily accessible and minimal time-invested therapy that could potentially alleviate nursing burnout and increase quality of care. methods participants four outpatient nursing units in the general internal medicine department at mayo clinic, rochester, mn were selected for participation in this study. the four nursing units were selected based upon the nurse population that worked full time, did not work in other units, and were able to participate in all five treatments. this selection criterion for the nursing units was applied to ensure that nurses would not see other therapy dogs at different frequencies, but that the nurses would be available during the visits. the nursing units within the department of general internal medicine included 1) women’s health and consulting medicine; 2) home enteral nutrition and home paracentral nutrition; and 3 and 4) two different departments of executive health. each nursing unit had six nurses participating in the study. a total of 24 nurses participated, 23 women and 1 man, with an average age of 43.13 ± sd = 11.76 years with 20.29 ± sd = 11.78 years of experience (table 1). all nurses participating in the study signed a consent form and a health insurance portability and accountability act (hipaa) waiver before completing any questionnaires or visiting with any therapy dogs. page 9 impact of therapy dog visits on nurses creative commons license 4.0 – non commercial – share alike – attribution clark, smidt & bauer https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science therapy dogs four therapy dog teams from mayo clinic, rochester’s caring canine program volunteered for this study. each therapy dog team is an active volunteer, seeing patients at the clinic regularly, registered with a therapy dog organization, and were willing to stop additional therapy dog visits for the duration of this study. the demographics for the four dogs were two males, a 12 yr old bichon (dog 1) and a 10 yr old small mixed breed (dog 2); and two females, a 7 yr old standard poodle (dog 3) and a 10 yr old shi-poo (dog 4). both male dogs had female handlers and were registered with pet partners. in contrast, both female dogs had male handlers; however, dog 3 was registered with therapy dog international and dog 4 was registered with alliance of therapy dogs. mayo clinic rochester’s iacuc committee approved this study (protocol a00003246-17). page 10 www pet behaviour science org creative commons license 4.0 – non commercial – share alike – attribution 2018 | vol.6 | 8 15 table 1. sociodemographics of the out-patient nurses. bdc, breast diagnostic clinic; exec, executive health; hen, home enteral nutrition program; hpn, home parenteral nutrition program; whc, women’s health clinic. *therapy dog that visited out-patient nurses for the duration of the project. study design the study was designed as a controlled before-after study for the nursing units experiencing therapy dog visits. the four therapy dog teams were randomly assigned to a nursing unit, by being asked to pick an envelope that contained the nursing unit they would be visiting. the therapy dog teams were then randomly assigned to the order of the four treatments using proc plan block randomization design on sas version 9.4. the treatments included treatment (trt) a, two visits a week for four weeks; trt b, one visit a week for four weeks; trt c, two visits over four weeks; trt d, one visit over four weeks; and trt e, control, no therapy dog visits. each visit lasted between 15 ± sd = 5 minutes in length and took place between 1130 and 1330. there was a one week washout period in-between each block. therapy dog team visits were consistent on time and day of the week for each team, but due to volunteer basis, it varied between teams. dog 1 and 2 visited on mondays and fridays, dog 3 visited on tuesdays and thursdays, and dog 4 visited on mondays and wednesdays. during this visit, the handler and dog met the nurses at a designated, quiet area. the session consisted of the nurses petting the therapy dog and conversing with the dog handler; visits were non-goal oriented. throughout the duration of the study, the therapy dogs were not allowed to make any additional hospital visits, to control for the exact frequency of visits during the four week period. prior to the start of therapy dog visits, nurses were asked to fill out a general demographic survey, the pet attitude scale-modified survey, and the lexington attachment to pet scale survey. during preand postvisits, participants were asked to fill out the maslach burnout inventory (mbi)―human services survey, the nursing workplace satisfaction questionnaire, the nursing work index―revised questionnaire, and a visual analog scale. statistical analysis the surveys among nursing units were analyzed as a completely randomized design using the mixed procedure of sas version 9.4 (sas institute inc., cary, nc). pre and post-survey scores for each different treatment were analyzed using a t test of sas 9.4. significance was set at (p ≤ 0.05), and the tendency was set at (p ≤ 0.10). results nursing demographic the 24 nurses that participated in this study consisted of 16 (66.66%) who were married, 2 (8.33%) divorced, 4 (16.66%) single, and 2 (8.33%) who chose not to answer. out of the participants, only one marked “yes” to smoking and 14 (58.33%) noted that they consuming 1 to 3 alcoholic beverages a week. two out of the 24 participating nurses did not own pets. when asked, 15 (62.50%) of participants did not have any intentions of leaving their nursing unit, 1 (4.16%) intended to leave their job within 12 months, and 8 (33.33%) were unsure. pet attitude scale-modified and the lexington attached to pet scale prior to the start of the treatments, the outpatient nurses were asked to complete the pet attitude scalemodified and the lexington attachment pet scale to obtain a generalized picture of how the participants felt about pets. nineteen (79.16%) participants responded with, “strongly agree” when asked if they loved pets and strongly agreed that house pets add to their happiness in life. in addition, 21 out of 24 (87.50%) strongly agreed that they would like to have a pet in their home; and strongly disagreed that having pets was a waste of money. in addition, 100% of the participants disagreed when asked if they hate animals. lastly, 21 out of the 24 (87.50%) participants, “strongly disagreed” that they were not very attached to their pets, with the remaining three participants, “somewhat disagreeing”. visual analog scale the visual analog scale is a 10-point scale, with zero meaning the highest possible level of feeling happy, energetic, relaxed, and calm, with a positive overall emotional well-being. a score of zero also indicates none at all for parameters including restlessness, fatigue, anxiety, depressed mood, and feeling a high level of stress. thus, a reduction in post-treatment page 11 impact of therapy dog visits on nurses creative commons license 4.0 – non commercial – share alike – attribution clark, smidt & bauer https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science scores indicates a positive outcome. when comparing pre and post–self-reported responses of the visual analog scale, trt b had a significant decrease (t80 = -2.24; p = 0.02) on depression scores from preto posttreatment responses. in addition, trt b had a significant decrease (t79 = -2.00; p = 0.04) on overall emotional well-being scores. trt a was significant (t80 = -2.60; p = 0.01) at reducing the post–self-reported scores of happiness. when comparing numbers, more post-treatment responses were higher within trt c and trt d, with five out of 10; trt e had six variables out of 10 (table 2). in addition to the notable numeric comparison between treatments, there was a significant nursing unit effect on the responses (anova: f3,94 = 4.64; p < 0.05). maslach burnout index (mbi)―human services survey the survey question regarding personnel feeling drained at work was significantly different (t15 = 1.76; p = 0.04) pre and post-treatment for trt a. the most common response to pre-therapy dog visits was “once a week”; however, after two weekly visits from a therapy dog over 4 weeks, the post-treatment response was “once a month or less.” the responses to the statement, “working with people all day is really a strain for me,” was significantly different between trt a (t17 = 1.89; p = 0.03) and trt d (t17 = -1.27; p = 0.04) when comparing pre and posttreatment responses. trt a’s most common pretreatment response was “once a week.” after the 4 weeks of therapy dog visits, trt a’s most common response was “one a month or less.” in addition, trt d’s most common pre-treatment response was “a few times a year or less,” and after a single visit with a therapy dog over the course of 4 weeks, the responses were “once a month or less.” lastly, the responses for trt a’s pre versus posttreatment responses to the statements, “i feel frustrated by my job,” and “i feel i’m working too hard on my job,” were significantly different (t17 = 3.20; p = 0.02 and t17 = 1.74; p = 0.03, respectively). the post-treatment response by over half of the individuals was “a few times a year.” in addition, trt a’s post-treatment responses for feeling frustrated at the job were “never,” and “a few times a year.” both responses decreased from the severity of their responses before therapy dog visits. nurse working index―revised there was no significant difference among or within treatments for the nurse working index―revised questionnaire. however, there were a unit effects, with three out of the four staffing adequacy questions. “enough time and opportunity to discuss patient care problems with other” was significantly different among units (anova: f3,172= 3.50; p = 0.01). “enough registered nurses on staff to provide quality patient care” was highly significantly different among units (anova: f7,175= 6.99; p < 0.05). lastly, “enough staff to get the work done” was highly significantly different among units (anova: f7,178 = 5.71; p < 0.05). there was also a significant different in regards to nurse-physician relation when asked about collaboration (anova: f7,177 = 2.03; p = 0.05). for all significantly different unit effects, nursing unit two had the highest, most agreeable responses, while nursing unit four had the lowest, most disagreeable responses. page 12 www pet behaviour science org creative commons license 4.0 – non commercial – share alike – attribution 2018 | vol.6 | 8 15 discussion the responses in the pet attitude scalemodified and lexington attachment to pet scale confirmed that over 80% of the participants in the study believe that owning pets contribute to their happiness and having a pet in the house adds to their overall happiness in life. these results suggest that those who participated in the study had a pet that they were attached to and thought it added happiness to their life. thus, the authors conclude that the addition of a dog as stress relief event would not trigger a stress response. the visual analog scale analysis supports the theory that therapy dog visits can help reduce compassion fatigue. in trt a, in which a therapy dog visited twice a week for 4 weeks, a significant increase in the feeling of happiness was indicated. in addition, trt b, which utilized a therapy dog visit once a week for 4 weeks, was able to markedly reduce self-reported responses of depression and improved overall emotional well-being. consequently, in trt e, the control/no therapy dog visit group, the least amount of improvement in the nursing units’ visual analog scale was seen, with six parameters being self-reported as worse. in addition, trt c, in which therapy dogs visited twice total over 4 weeks, and trt d, in which therapy dogs visited just once over 4 weeks, numerically had an increase in five page 13 impact of therapy dog visits on nurses creative commons license 4.0 – non commercial – share alike – attribution clark, smidt & bauer table 2. preand post–self-reported visual analog scores for each treatment (the smaller the number means the best possible/none at all). trt, treatment; 1treatment a: therapy dog visit twice a week for 4 weeks; 2treatment b: therapy dog visit once a week for 4 weeks; 3treatment c: therapy dog visit twice over 4 weeks; 4treatment d: therapy dog visit once over 4 weeks; 5treatment e: control (no therapy dog visit). a significantly decreased from preto post-treatment scores (p < 0.05). b numerically increased post–self-reported scores compared to pretreatment scores (zero meaning none at all or best possible). https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science visual analog scale scores, indicating that these five parameters worsened from pre to post-treatment responses. trt a was able to make a substantial positive effect on the post-treatment responses of the mbi―human service survey, which supports the hypothesis that therapy dog visits can help combat nurses feeling drained after work, strained during work, and overall less frustrated about their job. these preliminary results support the theory that having staff stress-relief events in the form of therapy dog visits, roughly once a week, can help nurses cope with compassion fatigue and burnout. the nurse working index―revised questionnaire; however, did not show any substantial effects from treatments. in contrast, there was a unit effect for staff adequacy questions and nurse-doctor collaboration, with unit two having the most agreeable responses and unit four having the most disagreeable responses. this suggests that the same level of collaboration and staffing are significantly different and the unit the nurse works in could greatly affect their risk for compassion fatigue, burnout, and overall wellbeing. therefore, while therapy dog visits may be effective at reducing compassion fatigue and burnout, they may not be helpful in the satisfaction level of nurses and their relationships with physicians and other nurses. past literature supports our findings. studies have been conducted observing the beneficial relationship that animals have with humans (nimer and lundahl, 2007). it has been noted that visits from dogs can considerably reduce anxiety levels and improve individual coping ability, which are both useful tools in overcoming compassion fatigue and burnout (baker and dawson 1998; berget et al., 2008). one major limitation to this study is that the nursing population was strictly outpatient nurses who were not always available, due to schedule or vacation time, to attend the therapy dog visits. while researchers prescreened nurses who stayed in the same speciality area, worked full time, and we were willing to attend, finding a time and date that worked for the dog and handler and all the participating nurses in each unit posed a challenge. a second limitation was the nursing unit effect. the participants within each unit voluntarily joined the study. however, naturally, some units are always a high stress area and nurses may have had individual hardships within the unit during the study period. in addition to this, there is potential bias from using survey responses. while the questionnaires used in this study are validated, the authors recognize that the nurses’ perception can add bias and to strengthen the study, objective parameters such as, oxytocin, cortisol, and heart rate variability, could be included in future studies. conclusion this small, novel study sheds light on potential avenues to assist nurses in overcoming compassion fatigue and burnout; however, larger studies should be conducted for a longer duration of time and be distributed more evenly among nursing units. in addition, it would add another point of view to introduce this staff stress event to inpatient nurse populations. this study supports the hypothesis that visits, most effectively weekly visits, from a therapy dog, can help alleviate stress and feeling frustrated, drained, and overwhelmed from working in the nursing field. it would be interesting to take this study further to observe the mechanism behind this conclusion. acknowledgements this publication was made possible by the mayo clinic ctsa through grant number ul1tr002377 from the national center for advancing translational sciences (ncats), a component of the national institutes of health (nih). references aiken, l. h., clarke, s. p., douglas, m., and sloane, d. m. 2002. hospital nurse staffing and patient mortality, nurse burnout, and job dissatisfaction. journal of american medicine association 288(16): 1987-1993. doi:10.1001/jama.288.16.1987. aiken, l. h., cimiotti, j. p., sloane, d. m., smith, h. l., flynn, l., and neff, d.f. 2011. page 14 www pet behaviour science org creative commons license 4.0 – non commercial – share alike – attribution 2018 | vol.6 | 8 15 the effects of nurse staffing and nurse education on patient deaths in hospitals with different nurse work environments. medical care 49(12): 1047-1053. doi. 10.1097/mlr.0b013e3182330b6e. barker, s. b., and dawson, k. s. 1998. the effects of animal-assisted therapy on anxiety ratings of hospitalized psychiatric patients. psychology series 49(6): 797-802. beetz, a., julius, h., turner, d., and kotrschal, k. 2012. effects of social support by a dog on stress modulation in male children with insecure attachment. frontier psychology 3: 352. berget, b., ekeberg, o., and braastad, b. o. 2008. animal-assisted therapy with farm animals for person with psychiatric disorders: effects on self-efficacy, coping ability, and quality of life, a randomized controlled trial. clinical practice epidemiology mental health 4(9). nimer, j., and lundahl, b. 2007.animal-assisted therapy: a meta-analysis. anthrozoology 20(3): 225-238. shimizutani, m., odagiri, y., ohya, y., shimomitsu, t., kristensen, t. s., maruta, t. m. and iimori, m. 2008. relationship of nurse burnout with personality characteristics and coping behaviors. industry health 46: 326-335. vahey, d. c., aiken, l. h., sloane, d. m., clarke, s. p., vargas, d. 2004. nurse burnout andpatient satisfaction. medical care 42(2 suppl): ii57-ii6. page 15 impact of therapy dog visits on nurses creative commons license 4.0 – non commercial – share alike – attribution clark, smidt & bauer https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science the health and welfare of dogs belonging to homeless people. david leonard williams and sarah hogg1 highlights dogs owned by homeless people are generally healthy with few behaviour problems. even though lower body condition scores have been found, only one dog was found to be underweight behaviour is not generally an issue in homeless peoples' dogs pet behaviour science | 2016, vol. 1, 23 – 30 david leonard williams and sarah hogg 1. department of veterinary medicine. university of cambridge paper research email: dlw33@cam.ac.uk united kingdom keywords: dog; health; homeless; pet; welfare 1. introduction homelessness affects many thousands of people in the uk and is a growing problem. reliable statistics are difficult to access as these individuals are often difficult to reach, moving between temporary accommodation and the streets (fitzpatrick et al. 2000). in autumn 2015, it was estimated that there were 2700 rough sleepers on the uk streets, with 21% of the national total bedding down in london (thames reach, 2015). in addition there are hundreds of people without a permanent home sleeping in hostels, or in sub-standard or temporary accommodation. homelessness has significant negative impacts both upon those who are homeless and the society which renders them thus (communities and local government. 2011; sakelaropoulos et al. 1998). they are at greater risk of poor health outcomes (o’grady and gaetz 2004; frankish et al. 2005) and increased use of mental health and addiction services than homed individuals (caton et al. 1995; grimm and maldonado page 23creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 23 30 abstract a significant number of homeless people own dogs, with these animals contributing to the well-being of their owners by providing emotional support and in many cases, a reason for living as well as acting as what might be termed a social catalyst, improving bonds between their owners. yet many consider that homeless people should not be allowed, let alone encouraged to keep a dog. they consider that living with homeless people must have a negative impact on the dog’s health and welfare compared to that of a dog owned by people with a home. here we sought to determine the health and welfare of dogs owned by homeless people, comparing 50 dogs owned by homeless people with 50 owned by people living in a home. in contradistinction to the negative view noted above, we found that dogs owned by homeless people were healthy animals, less likely to be obese, had fewer behaviour issues such as aggression to strangers and separation anxiety when compared to dogs owned by people living in a conventional home. we suggest that these findings should be taken into account when deciding whether a homeless person with a dog should be allowed into a hostel, and indeed the general attitude of the public to homeless people living with a dog by their side on the street. http://www.petbehaviourscience.org/ 1995; bogard et al. 1999; burt et al. 2001; taylor et al. 2004). it is not just that homeless people lack the physical protection which a house affords; their predicament is a destructive experience leaving them without social and legal dimensions, roots and identity. they are often bereft of a sense of belonging and the emotional well-being which living in a home provides (crisis, 2011, homeless link. 2009). the human-animal bond promotes the welfare of both parties (davis and balfour 1992, russow 2002, labreque and walsh 2011) with the physical and emotional benefits of pet ownership being well documented (edney1993; serpell 1991; wilson and turner 1998; scas 1999; staats et al. 1999). dogs can act as a ‘social catalyst’ and provide the emotional support and social relationships often lacking in homelessness (robinson 1997; rew 2000). dogs are the most popular pets belonging to the homeless, benefitting homeless people by facilitating communication with others (menteith 2001; kidd and kidd 1994), combating the intense feelings of isolation that homelessness can give. dog ownership can be a vital method in coping with suicidal thoughts (rew 2000) and reducing drug use common amongst the homeless (baker 2001) probably through fear of imprisonment which would necessitate relinquishing the dog (taylor et al. 2004). around one third of the homeless population turn to crime, with minor offences such as shoplifting being common (ballintyne 1999). dog ownership appears to reduce the likelihood of a homeless person committing a crime (taylor et al. 2004) again potentially because imprisonment necessitates loss of the animal. crucially, dogs give their owner a sense of responsibility; in 2000, rew found that they inspired better decision making amongst young homeless people. taylor, williams and gray (2004) found that homeless people are strongly attached to their pets and depend upon them for companionship, stability and security, and showed that the homeless were generally more empathetic towards animals than the general public. having made all those positive comments, it is a sad fact that the majority of homelessness projects in the uk do not accept or admit dogs (dogs trust project 2007) although a growing number are now allowing pets to be accommodated. although they badly need the support of such services, many homeless people are unable to receive help simply because they own a dog (singer 1995). programme facilitators are often concerned about dogs’ potential behavioural problems, such as toileting and aggression and perceived health issues such as fleas and worms (hope project 2015). for the homeless, many of whom have experienced multiple forms of loss, being forced to give up their pet in order to access accommodation may cause further emotional trauma (labrecque and walsh 2011). the majority of homeless dog owners would rather remain homeless than accept accommodation that would not allow their pets (hart and zasloff 1995; baker 2001). a dog, therefore, can be the limiting factor that keeps a homeless person on the streets permanently (masters 1998; menteith 2001). there is little information in the literature concerning homelessness and companion animals (labrecque and walsh 2011). whilst the primary focus of such literature has rightly been the welfare and health of the owner, there is a lack of literature regarding the health or welfare of the dogs themselves. here we present the results of a clinical health examination of 50 dogs owned by homeless people and 50 owned by settled individuals together with details of discussion with the owners on the health and welfare of their pets. 2. methods questionnaire and ‘health check’ examination a purposive convenience sampling technique was used to recruit dogs and their owners for a standardized clinical ‘health check’ examination and questionnaire regarding the dog’s care. 50 homeless dog-owner pairs were approached whilst on the street in cambridge and london; 38 non-homeless participants were approached whilst walking their dogs in the same sampling areas and 12 whilst presenting their healthy dogs for vaccination or post-neuter consultations at the rspca clinic, run by the veterinary school in cambridge. all homeless people with dogs encountered while walking the streets of these two cities were asked whether they would be prepared to take part in the study. without exception they agreed after the reasons for the survey were explained and time taken to gain their trust and confidence. most page 24 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 23 30 seemed happy that positive attention was being paid to them and their pet. similarly homed owners walking their dogs were generally happy to be involved with only three people saying they were too busy to stop and talk. owners were asked the questions noted in table 1; their dog’s age, how long they had owned the dog and its source, about its diet and exercise regime, and any routine veterinary care received. the owners gave a verbal consent to the dogs being examined after which the dogs were then given a brief clinical examination following a sheet detailing each area to be examined (figure 1) concluding with a conversation with the owner where any problems were discussed. the dog's body condition score was assessed using standard techniques previously reported (laflamme 1997) both visually and by palpating how easily the ribs can be felt, as shown on the world small animal veterinary association website (wsava, 2013). rectal temperature was not taken but cardiac auscultation and abdominal palpation was possible. what was the source of your pet? how long have you had the animal? what do you feed your pet? how much exercise do you estimate the dog gets daily? how would you describe your dog’s character? does your dog have any behavioural issues? does your dog have any physical illnesses? does your dog get regular veterinary attention? is your dog regularly vaccinated and dewormed? table 1. questions asked of owners during informal discussion finally, the dogs' demeanour was noted as being friendly, outgoing and approachable, neutral, shy and withdrawn or overtly aggressive and owners were asked about any behavioural problems that their dog showed. interviews were conducted with a technique sensitive to the emotional perspective of homelessness. the study received ethics approval from the university of cambridge veterinary school ethics committee. all participants gave informed consent. no reward was offered for participation in either health checks or the survey, and responses throughout were anonymous. 3. results a significant difference was found between the ages of dogs belonging to homeless and non-homeless individuals (homed: mean±standard deviation 5.4±3.98, n = 50; homeless: 3.7±2.9, n = 50, p = 0.03) with dogs belonging to homeless people being significantly younger. no homeless-owned dogs surveyed was greater than 12 years of age. source of dog, and length of time in owner’s possession a considerable difference in reported sources of dogs was noted, with 80% of dogs belonging to settled owners being purchased compared to only 4.5% of dogs belonging to homeless people (p<0.01). all of these purchased dogs were purchased before the owners became homeless. whereas the great majority of non-homeless-owned dogs had been with their owners since 8 weeks of age or earlier (70%) and were purchased from private sellers or breeders (66%), most homeless-owned dogs were acquired later in life (70%) and were reported to be strays (39%) or acquired from friends, several of whom had died or were going into prison (27%). 69% of dogs belonging to settled owners were in multi-pet household, whereas 81% of dogs belonging to homeless owners were the only pet (p<0.05). diet whereas most settled dog owners used a commercial, branded dog food as the main constituent of their dogs’ diet (38/50), the majority of homeless dog owners chose a supermarket own-brand food (36/50). whilst 95.5% of homeless owners fed their dogs a dry diet, only 46% of settled owners chose a predominantly dry diet, with the remainder either feeding a mixed, or entirely wet diet. exercise many homeless people found it hard to quantify how much exercise their dogs get in a day, as they are out and about being walked for several hours. 77% of homeless owners mentioned extra walks or specific park visits. dogs are often kept off their leads, and so are free to wander as they wish. 88% of dogs belonging page 25 dogs belonging to homeless people creative common license 4.0 – non commercial – share alike – attribution williams, d.l. and hogg, s. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science to settled owners, however, were subject to a twice daily routine of walks, usually lasting 20-30 minutes. 22% of owners reported that this varied with work or social commitments. the remainder of dogs are not walked daily, or are kept in the house permanently. demeanour dogs belonging to homeless people were very significantly more likely to be described as ‘quiet, but friendly’ (χ2 = 10.3, df = 1, p = 0.001), whereas those belonging to settled people were more likely to display aggression towards the person giving them a health check (χ2 = 4.65, df = 1, p = 0.031). dogs belonging to page 26 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 23 30 figure 1 physical examination sheet settled owners were also significantly more likely to be hyperactive or openly disobedient during the interview (χ2 = 3.68, df = 1, p = 0.055). no significance was found for dogs being friendly and moderately outgoing (χ2=1.61, df = 1, p = 0.205), friendly and very outgoing (χ2=0.865, df = 1, p = 0.352), nervous and withdrawn (χ2= 1.31, df = 1, p = 0.284) or defensive of their owner (χ2= 0.491, df = 1, p = 0.484). behavioural issues homeless-owned dogs were significantly less likely to have behavioural problems reported than settledowned dogs (χ2 = 24.2, df = 1, p < 0.001). dogs belonging to settled owners were significantly more likely to display inappropriate barking (χ2 = 9.2, df = 1, p = 0.002) or chewing and destructive behaviour (χ2=7.60, df = 1, p = 0.006), and significantly more likely to have toileting issues (χ2 = 5.02, df = 1, p = 0.025). no significance was found for incidents of aggression (χ2=2.63, df = 1, p = 0.105), separation anxiety (χ2 = 2.42, df = 1, p = 0.102), excessive nervousness (χ2 = 0.476, df= 1, p = 0.494), pulling on the lead or showing frequent attention seeking behaviour (χ2 = 2.73, df = 1, p = 0.099, for each category). physical health a significant difference was noted between the body condition score (bcs) of dogs belonging to the homeless and non-homeless communities (homed: 5.7±1.23, n = 50; homeless: 4.3±0.97, n = 50, p = <0.01) where 5 is considered an acceptable bcs and as value above 5 as over-weight (laflamme 1997). homelessowned dogs had a significantly lower bcs, but spread over a healthier range than for non-homeless-owned dogs, which have a far higher chance of being overweight or obese. there was no significant difference between the numbers of physical abnormalities found in homelessowned or settled owned dogs (χ2 = 0.713, df = 1, p=0.933), nor were there any significant differences found for the following conditions: skin and ear problems (χ2=0.094, df = 1, p = 0.889), ectoparasites (χ2=2.42, df= 1, p = 0.102), wounds, serious scarring or infected feet (χ2 = 3.3, df = 1, p = 0.069), gastrointestinal problems (χ2 = 2.34, df = 1, p = 0.126), cardiac problems (χ2 = 2.73, df = 1, p = 0.099), arthritis and other locomotor problems (χ2 = 3.21, df = 1, p = 0.073), congenital defects (χ2 = 3.68, df = 1, p = 0.055), senile or degenerative changes (χ2 = 0.713, df = 1, p = 0.933). routine veterinary procedures registration and regular access to veterinary care was far higher amongst settled dog owners, with 95.5% reporting routine use of a veterinarian. 15 of the 42 dogs cited a charity clinic, as opposed to a private veterinarian, as their usual provider of veterinary care. amongst homeless participants, 27% reported no access to veterinary care, 46% stated that although they didn’t regularly access veterinary care they would use a charity clinic were it needed, and 27% named a clinic or facility with which they were a registered, regular client. dogs belonging to settled owners were more likely to be vaccinated with 78% of dogs having up-todate cover, as opposed to 39% of homeless dogs (p<0.01). 16% of dogs belonging to settled owners were reported to have recently lapsed vaccinations, compared to 18% of homeless-owned dogs. only 3 out of 50 settled owners chose not to vaccinate, compared to 18 of 44 homeless owners. 80% of settled owners reported regularly worming their dogs, 6% of having wormed but with lapsed treatments and 14% deciding not to treat their dogs; compared to 30%, 30% and 40% of homeless owners respectively. microchipping was reported to be a more common practice amongst settled owners with 62% microchipping their dogs, compared to 22% of homeless owners. many homeless owners cited a lack of fixed address as an obstacle to useful microchipping. several homeless owners who had microchipped their dogs had done so before becoming homeless, and now had the incorrect contact details on record. 50% of settled owners reported regularly treating their dogs for fleas, 10% having given treatments which were now overdue and 40% deciding not to treat; compared to 27%, 25% and 48% of homeless owners respectively. 4. discussion contrary to much opinion, it does not appear from this study that dogs belonging to homeless people are less healthy than those belonging to settled owners, although they do predominantly experience a different page 27 dogs belonging to homeless people creative common license 4.0 – non commercial – share alike – attribution williams, d.l. and hogg, s. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science range of health conditions unique to the conditions in which they live on the streets. this is not what the public would seem to believe, with most people expecting dogs belonging to homeless people to be in poorer health than those belonging to the nonhomeless. aside from fleas, skin complaints affecting homelessowned dogs tended to be calluses, and other physically induced conditions, whereas non-homeless-owned dogs tended to suffer from allergy, atopy and masses such as histiocytomas and lipomas. gastrointestinal complaints in homeless dogs tended to be ownerreported diarrhoea. there is a high chance this could be caused by worms, but could also be due to the higher probability of dietary indiscretion or an irregular diet. the only gastrointestinal problem reported in the nonhomeless population was motion sickness; not a complaint that homeless dogs are likely to suffer from as they rarely, if ever, travel in vehicles. although the statistics showed the numbers of these dogs as insufficient to provide a significant difference between populations, dogs with cardiac, endocrine and senile or degenerative changes were found in the homed, but not the homeless population. this may be because dogs with severe cardiac and endocrine problems cannot survive on the street and dogs on the street do not live to be old enough to develop problems related to senility. the population of dogs belonging to homeless people would appear to have different characteristics than that belonging to settled owners. the majority are strays, as opposed to purpose-bred and purchased, and the population has a younger average with dogs at the oldest end of the spectrum (over 12 years of age) missing entirely from our population sample. in homeless people, the average life expectancy is just 42 years of age (crisis 2002), compared to 78.0 years for settled males and 82.1 years for settled females in england (ons 2010). the cold, damp, unsanitary conditions they are forced to live in leave them more susceptible to disease and injury. if such conditions have such a drastic impact on human life expectancy, then they are likely to impact on animals kept in the same way. dogs belonging to the homeless in this study had significantly fewer behavioural problems than those belonging to settled owners. several factors could explain this. dogs belonging to the homeless have a much greater range and degree of contact both with people and other dogs, so are better socialized. they receive more exercise than dogs belonging to settled owners, tending to be more constantly wandering with additional park visits to allow them to run around freely, as opposed to subject to a two-outings-a-day regime which is subject to their owners work and social commitments. as they are not frequently left alone for long periods of time, they do not suffer from separation anxiety, and as they are usually the only pet or companion, they receive plenty of individual attention and mental stimulation. only just under a third of the survey participants supported this result. homeless dogs had a significantly lower bcs than the non-homeless population. however, only one dog was found to be underweight, and this was matched by a similar dog amongst the housed population. many dogs in the homed population were overweight, or obese. homeless dogs were healthier in this respect, which may be because they receive more exercise, are less likely to be overfed or given calorie-rich food because of financial reasons, or because living outside in the cold means they have a higher basal energy demand. clearly examining the animals on the street places limits on the degree of evaluation possible but physical examination including thoracic auscultation, abdominal palpation, assessment of pulse and respiration are all possible to give a reasonably detailed clinical examination close to what might be possible in a veterinary consulting room. other issues involve the impossibility of performing ancillary tests. without the use of faecal egg counts, for example, it was impossible to screen dogs for endoparasites and without the opportunity to perform skin scrapes for mites, wet paper tests for flea dirt or cytological impression smears, the presence of ectoparasites was difficult to ascertain. the inability to evaluate faecal samples for worm egg counts similarly made detection of gastrointestinal parasitic disease. this would be useful in future extensions of this research to confirm or disregard gastrointestinal page 28 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 23 30 parasites as the causes of reported diarrhoea cases, and to test the hypothesis that endoparasites are more common in dogs belonging to the homeless. additionally, although dogs suffering from symptomatic cardiac problems as noted from auscultation, demonstration of a pulse deficit and a history of poor exercise tolerance were noted in a small number of homed dogs, it is difficult to fully evaluate a dog’s cardiac status, or to detect subtle abnormalities, on the street. another significant issue concerns the fact that it is difficult when interviewing people about the care of their pet to be sure of an entirely truthful answer. whilst many settled owners were happy to chat, particularly in the veterinary surgery at the rspca, about their animals’ veterinary problems, homeless participants were sometimes reluctant to admit such issues. understandably, they are wary of people wishing to document what may be seen as neglectful on their part. it is difficult for the interviewer to build up necessary trust within a short time frame. settled owners sometimes seemed to exaggerate the amount of exercise their dog receives, as they also did not wish to be perceived as neglectful. behavioural issues seemed to be under-reported, with owners telling anecdotes where the dog exhibited a problem, or with dogs sometimes exhibiting behavioural problems during the health check, followed by the owner reporting no problems with their dog. in such cases, issues were noted as part of the health check. literacy weaknesses are inevitable with most questionnaires, leading to varying interpretations of questions. this may have been an issue with the electronic survey, although the opportunity to directly contact the researchers, or leave a comment at the end helped to alleviate this. whilst working with the homeless community, a more phenomenological approach and more flexible interview questions may be useful to allow interviewees to explain their choices (e.g. diet, access to veterinary care) more formally. an ethnographic technique would be the most useful method, in order to ensure a good basis for trust. for any future work, it would be useful to work in partnership with shelters and/or outreach schemes in order to survey a wider number of homeless pet owners, and to perform health checks in more amenable surroundings. offering worming medication, flea treatments, food or toys may be beneficial to dogs of participating homeless owners. 5. conclusion the purpose of this study was to explore the health status of dogs belonging to homeless people; an area that has received very little attention in scientific or sociological literature. the key findings demonstrated that dogs belonging to the homeless are not significantly less healthy than those belonging to nonhomeless individuals. they have lower body condition scores, but this is because the non-homeless dog population tends towards being overweight or obese. they also appear to have fewer behavioural problems. further investigation into the health of dogs belonging to homeless people would be invaluable in improving homeless-accessible veterinary services, improving public education and counterbalancing the negative stigma often attached to homeless pet owners. it would also raise awareness of professionals in the field who are often concerned about dogs’ behaviour and health issues that making their facilities pet-friendly is not tantamount to inviting in a human health risk, or viable criticism that they are supporting owners who do not properly care for their animals. these dogs do not exhibit behaviour that is likely to discourage other users of such services from doing so. dogs can be a valuable asset to the homeless, providing a friend and companion in what may otherwise prove an isolating and trustless way of life. it is not acceptable that they can prevent a homeless person from accessing health care or finding accommodation. 6. references baker, o. 2001. a dog’s life. homeless people and their pets. oxford: blue cross ballintyne, s. 1999. unsafe streets: street homelessness and crime. london: institute for public policy research communities and local government. 2011. statutory homelessness: march quarter 2011 england (isbn 9781409825630) page 29 dogs belonging to homeless people creative common license 4.0 – non commercial – share alike – attribution williams, d.l. and hogg, s. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science crisis. 2002. statistics about homelessness. london: crisis crisis. 2011. statistics about homelessness. london: crisis davis, h., balfour, d. 1992. the inevitable bond: examining scientist-animal 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https://www.moretodogstrust.org.uk/downloads/happy-&-healthy-2007-final.pdf http://www.wsava.org/sites/default/files/body%20condition%20score%20chart%20dogs.pdf http://www.wsava.org/sites/default/files/body%20condition%20score%20chart%20dogs.pdf http://www.thamesreach.org.uk/news-and-views/homelessness-facts-and-figures/ http://www.thamesreach.org.uk/news-and-views/homelessness-facts-and-figures/ http://www.ons.gov.uk/ons/rel/health-ineq/trend-in-life-expectancy-by-socioeconomic-position-by-the-national-statistics-socioeconomic-classification--england-and-wales/1982-86-to-2007-11/stb-trend-in-life-expectancy.html http://www.ons.gov.uk/ons/rel/health-ineq/trend-in-life-expectancy-by-socioeconomic-position-by-the-national-statistics-socioeconomic-classification--england-and-wales/1982-86-to-2007-11/stb-trend-in-life-expectancy.html http://www.ons.gov.uk/ons/rel/health-ineq/trend-in-life-expectancy-by-socioeconomic-position-by-the-national-statistics-socioeconomic-classification--england-and-wales/1982-86-to-2007-11/stb-trend-in-life-expectancy.html http://www.ons.gov.uk/ons/rel/health-ineq/trend-in-life-expectancy-by-socioeconomic-position-by-the-national-statistics-socioeconomic-classification--england-and-wales/1982-86-to-2007-11/stb-trend-in-life-expectancy.html the human response to pet rescue tv commercials 2020 vol. 09 34 44 www.petbehaviourscience.com the human response to pet rescue tv commercials thomas mueller*, twila wingrove and sidney murray abstract: according to the american society for the prevention of cruelty to animals, 6.5 million abandoned pets are housed in shelters in the united states. of those who are sheltered 670,000 dogs are euthanized. an opportunity exists to reduce the number of euthanized animals, primarily through adoption. several benefactors to this animal rescue initiative have prompted pet rescue adoption through television commercials. this study explored the perception of viewer“attitude toward the ad” and“intent to adopt” for a pet rescue television commercial. study respondents (n=335) watched “somebody to love” a commercial featuring a lonely man and his emotional attachment with an abandoned dog. gender identity was significantly related to attitudes toward the commercial. participants identifying themselves as feminine held more favorable attitudes towards the commercial, whereas participants identifying as more masculine reported less favorable attitudes towards the commercial. work status and ethnicity were also significant predictors of attitude toward the ad. non-workers were more highly responsive to the commercial, than were students or those in working capacity. participants identifying as any ethnicity other than white held less favorable attitudes towards the commercial, which features white actors. this implies it is important to examine commercial content regarding plot and context, as the viewer response is affected by gender, ethnicity and employment status. producers of future animal rescue commercials should consider content specific to alternate human identities, where rescue animals can facilitate a common compassion among potentially discordant identity groups. thomas mueller1*, twila wingrove2, sidney murray2 1 department of communication appalachian state university 121 bodenheimer drive boone, north carolina 28608 2 department of psychology appalachian state university 112c smith-wright hall boone, north carolina 28608 *correspondence author: department of communication appalachian state university 121 bodenheimer drive boone, north carolina 28608 e-mail: muellerts@appstate.edu phone: (828)262-2228 highlights creative common license 4.0 – non commercial – share alike – attribution page 34 keywords: animal adoption; animal euthanization; pet adoption; pet advertising; pet rescue. • this paper presents a case study on animal rescue through television commercials. • researchers can assist pet rescue organizations in developing media messages that affect the highest response rates among viewers. • media promoting pet rescue must be sensitive to alternate ethnicities and pet behaviour science 2020, vol. 9, 34 44 doi:10.21071/pbs.vi9.12193 mueller, wingrove & murray thomas mueller, twila wingrove, sidney murray pet behaviour science introduction according to the american society for the prevention of cruelty to animals, 6.5 million abandoned pets are housed in shelters in the united states. the largest proportion of animals are 3.3 million dogs and 3.2 million cats. of those who are sheltered 1.5 million are euthanized (670,000 dogs and 860,000 cats). if any part of this statistic can be considered positive, it is that the number of euthanized animals has decreased from 2.6 million in 2011 (“aspca on pet statistics: how many pets are in the united states? how many animals are in shelters?,” n.d.) an opportunity exists to reduce the number of euthanized animals, primarily through adoption. some countries, such as italy, have enacted laws that do not allow euthanizing animals. without the option of euthanizing, temporary adoption programs have been devised. animals are matched to care givers based on breed and size of animal desired (normando et al., 2006). dotson and hyatt (2008) canvased 749 dog owners to expose underlying dimensions that are part of the dog and human companionship model. the study concluded that gender, age and education were related to the highly reliable association between a pet and owner, which leads to emotional, physical and mental health. the pet-human relationship is also affected by quality time with the dog, length of association with the dog, and whether the dog is identified as mixed or purebred. several studies have explored the likelihood of an adoption. for example, animals who stay in the front portion of the kennel, do not bark, and gaze into the eyes of potential adopters increases the chances for adoption. this study concluded animals that ignored play initiation negatively affected adoption choice, while lying in proximity to the adopter positively affected the adoption choice (protopopova & wynne, 2014). mondelli et al. (2004) indicated behavioral problems can emerge, exacerbated by lack of a yard or garden, or outdoor terrace. some animals are adopted, but are eventually returned to their shelter of origin. as a part of an initiative to increase the number of rescued pets, sponsors of pet rescue organizations have employed marketing messages to heighten the public awareness for lonely and abandoned animals. a study by phillips (1996) indicated when animals are featured in commercials, they can assist in setting the cultural context for products and services. some attachment to animals in television commercial settings has been likened to a consumer consumption experience. for example, people relate to unique benefits depicted in a product or service and respond to a call for action, which is to move towards purchase. people relate to the pets, see benefit in pet interactions, which leads to pet adoption as a perceived positive outcome. pets are welcomed into our most private endeavors, elevated to the status of children or friends. the unconditional love offered by pets make these consumption experiences meaningful in our actions and lifestyles (holbrook, stephens, holbrook & strazar, 2001). associations between animals and society currently exist in culture and those associations can influence product interpretations. when companionship with their dog is a high priority, dog owners can experience emotional wellbeing, lower stress and reduce blood pressure. it has been suggested human attraction to canines is sacred, is in the creative common license 4.0 – non commercial – share alike – attribution page 35 cultures. context and storytelling around the animal and its relationship with humans becomes critical. pet behaviour science 2020, vol. 9, 34 44 doi:10.21071/pbs.vi9.12193 moment and can lead humans into a different level of interaction (maharaj, kazanjian, & haney, 2016). to develop a strong response and success rate in pet rescue, it is essential to understand the motivators to adopt pets within the target audience. societal shifts and cultural change within industrialized democracies has altered how humans view the animals they intend to rescue. markovits and queen (2009) explored the differentiations among gender within a canine rescue group. individuals who identified themselves as females saw their commitment to animal rescue related to raising social capital. it reinforces ties to like-minded humans, more than creating opportunities for new relationships with people holding alternative values. the study concluded that females involved with dog rescue are part of a positive association process. earlier studies indicated dog rescue among females was an effort to alleviate loneliness due to a lack of social contacts (katz, 2003). females may act as dog rescuers as part of a contribution to the social good in society. men in this study held a perception that women had more time, and fewer responsibilities, which affected their ability and desire to adopt pets. animals in commercial and advertising settings are frequently focused on in consumer behavior research. amyx (2017) identified subjects demonstrating high engagement with dogs and cats as pets. dogs and cats as pets were portrayed in a commercial that used an experimental setting. both cat and dog lovers preferred their respective advertisement, preferred the realtor depicted in the advertisement and held higher purchase (adoption) intentions. a scale used in the study indicated cat lovers are motivated by individualism and independence. dog lovers are motivated by dependability and security. however, is important to study gender effect on pet rescue. ramirez (2006) examined pet owners, gender norms, and development of relationship with their dogs. gender norms are correlated with the perception of what is a suitable dog, definition of animal behavior through gender identity, and use of dogs to promote one’s gender identity. for example, dogs have been historically identified as masculine while cats depict a feminine nature. respondents in this study indicated cat ownership is well known as a favorite species among gay men. when audience segmentation is further defined, gender identity reflects a sense of self within social expectations, roles and behaviors. the loosening of gender stereotypes has become a part of social conversations related to experiential identity and lifestyle options. the millennial generation has embraced a gender spectrum where gender fluidity might help individuals feel more authentic yet may cause confusion based on how interaction should occur. parker (2016) suggests society should strive to honor diversity and individuality while avoiding bias, prejudice or violence. this study contributes to the potential for higher survival rate among abandoned animals by exploring consumer response to television commercials promoting animal adoption. personal identities related to gender, spirituality, political association and other key demographic variables were measured and tested as predictors of response to pet rescue commercial appeal. based on the literature that suggests a differentiation among how humans perceive their animal experience, the hypothesis of this study was that the demographic and psychographic differentiation among consumers will produce an effect on the attitude towards the “lonely man” pet rescue television commercial. creative common license 4.0 – non commercial – share alike – attribution page 36 thomas mueller, twila wingrove, sidney murray pet behaviour science 2020, vol. 9, 34 44 doi:10.21071/pbs.vi9.12193 2020 vol. 09 34 44 www.petbehaviourscience.com methods study design & participants this study was conducted as part of a state university research course. eighteen students participated as co-investigators. each student attained accreditation from the collaborative institutional training initiative (citi) as part of the institutional review board (irb) research approval process. data were collected over a 14-day period using the online survey software qualtrics. students identified potential respondents using convenience and judgmental sampling techniques, qualifying respondents who currently had a pet in the home or had cared for a pet in the past. after eliminating 39 cases with incomplete survey responses or a lack of informed consent, 335 participants were retained for the final sample. demographics are reported in the results section. materials commercial video. the animal rescue commercial featured in this study was created by agency siltanen & partners for client coldwell banker, a residential real estate franchise system. coldwell banker utilizes a strategy that frames emotion, rather than the transaction of home buying, in promoting its real estate brand (stanley, 2017). the advertisement anchored a creative campaign that supports the firm’s three-year partnership with adopt-a-pet.com. it was reported that the association between coldwell banker and adopt-a-pet.com facilitated more than 20,000 pet adoptions. the commercial is entitled “somebody to love.” it depicted a sad, isolated man and a stray dog, initially rescued from an alleyway. the animal used in the commercial, named “scout,” was an actual shelter pup. at the time of this writing, the commercial had over 271 million views (“coldwell banker tv commercial, ‘somebody to love’ ispot.tv,” n.d.) (table 1). the adopt-a-pet television campaign was supported by coldwell banker’s 84,000 real estate agents, who facilitated pet adoptions through relationships in their local communities. creative common license 4.0 – non commercial – share alike – attribution page 37 table 1. coldwell banker tv commercial attitude and intent to adopt. responses to the “somebody to love” advertisement were assessed through the 10-item advertising scale developed by spears and singh (2004). five items measured intention to purchase (adopt) and five items measured general attitudes toward the advertisement. all items used four-point semantic differential scales. for example, 1 was used to score “unappealing” and 4 was used to score “appealing.” the scale held internal consistency in this sample (α = .92) and was used as a unidimensional dependent variable (table 2). thomas mueller, twila wingrove, sidney murray pet behaviour science 2020, vol. 9, 34 44 doi:10.21071/pbs.vi9.12193 mueller, wingrove & murraypet behaviour science https://www.youtube.com/watch?v=oi_5-0z_jzu creative common license 4.0 – non commercial – share alike – attribution page 38 table 2. 10-item attitude and intent scale gender identity. a single item semantic differential 10-point scale was used to measure gender identity. the item was anchored with “feminine” indicating 1 to “masculine” indicating 10. participants were prompted with the following: “please help us understand your gender, mark your identity on the scale below.” spirituality. a single item semantic differential 10-point scale was used to measure participants’ degrees of spirituality. the item was anchored with “atheist” indicating 1 to “highly spiritual” indicating 10. participants were prompted with the following: “religious affiliation can be important to consider in our study. the scale below is labeled from atheist to highly spiritual. where would you place yourself on this scale?” political affiliation. a single item semantic differential 10-point  scale was used to measure political affiliation. the item was anchored with “alt-left” indicating 1 to “alt-right” indicating 10. participants were prompted with the following: “the political scale below is labeled from alt-left to alt-right. where would you place yourself on the scale?” work status. participants were prompted with a single item: “what’s your current work status?” they chose from the following options in a drop-down menu: “student don’t work,” “student + work,” “business executive,” “business manager,” “administrative work,” “employee,” “teaching and instruction,” “military,” “self-employed,” “unemployed,” “stay at home parent,” “retired,” and “other.” for data analysis, we created three categories: student, currently employed, and unemployed. income. participants were prompted with: “information about income is very important to understand. would you please give your best guess? please indicate the answer that includes your entire household income in (previous year) before taxes.” participants chose from the following drop-down menu options: “less than $10,000,” “$10,000 to $19,999,” “$20,000 to $29,999,” “$30,000 to $39,999,” “$40,000 to $49,999,” “$50,000 to $59,999,” “$60,000 to $69,999,” “$70,000 to $79,999,” “$80,000 to $89,999,” “$90,000 to $99,999,” “$100,000 to $149,999,” and “$150,000 or more.” in the analysis, income was broken into four groups: up to $29,999, between $30,000 and $59,999, between $60,000 and $89,999, and $90,000 and above. thomas mueller, twila wingrove, sidney murray pet behaviour science 2020, vol. 9, 34 44 doi:10.21071/pbs.vi9.12193 2020 vol. 09 34 44 www.petbehaviourscience.com analytical plan our primary research question was whether demographic variables were related to attitudes towards the commercial. therefore, we first evaluated the characteristics of the sample on these key demographic variables, then tested for relationships between each demographic characteristic with the dependent variables using t-tests, anovas, and correlations. finally, we tested the combined predictive effect of gender, spirituality, politics, work status, income, ethnicity, age, education, relationship status and dog owner status that were significantly related to commercial attitudes using multiple regression. results creative common license 4.0 – non commercial – share alike – attribution page 39 ethnicity: participants were prompted with, “what is your racial or ethnic heritage?” the subsequent answer choice options were: “non-hispanic white or euro-american,” “black, afro-caribbean, or african american,” “latino or hispanic american,” “east asian or asian american,” “south asian or indian american,” “middle eastern or arab american, “native american or alaskan native,” and “other.” due to small numbers of non-white participants, we created two groups: white participants and all non-white participants. age. participants were prompted with a single item: “please share your age with us.” they chose from the following drop-down menu options: “18-24,” “24-34,” “35-44,” “45-54,” “55-64,” “65-74,” and “75 years or older.” for the analyses, age was grouped by 18-34; 35-54; and 55+. education. a single item, “what is the highest level of education you have completed?” prompted participants. the following choices appeared on a dropdown menu: “less than high school,” “high school/ged,” “some college,” “2-year college degree,” “4-year college degree,” “master’s degree,” “doctoral degree,” and “professional degree (jd, md).” education was analyzed at three levels: less than an associate’s; associate’s or bachelor’s degree; and master’s degree and above. relationship status. the prompt: “what is your current relationship status?” was presented with the following answer choices: “single,” “married,” “cohabiting,” “widowed,” “divorced,” “separated,” and “other.” relationship status was analyzed at three levels: single; married or cohabiting; and widowed, divorced, or separated. “other” was excluded from the analysis. dog ownership status. to gauge whether participants have owned or currently own a dog, participants were prompted with: “i have a dog now, or had a dog in the past.” they chose from response options “yes” and “no” in a drop-down menu format. sample demographics are reported in table 3. ninety percent of the sample had owned a dog. the sample was mostly non-students on the lower end of income status. they were mostly white young adults with a college degree or less. the participants’ average scores leaned towards feminine, religious, and conservative. a strong majority were dog owners. next, we tested using correlation analysis to determine if each demographic variable was independently related to attitudes about the commercial. the only demographic variable that was significantly related to attitudes about the commercial was gender, r = -0.19, p < 0.001. participants identifying themselves as more feminine held more favorable attitudes towards the commercial, whereas participants identifying themselves as more masculine reported less favorable attitudes towards the commercial. neither political affiliation nor spirituality were related to commercial attitudes. thomas mueller, twila wingrove, sidney murray pet behaviour science 2020, vol. 9, 34 44 doi:10.21071/pbs.vi9.12193 mueller, wingrove & murraypet behaviour science creative common license 4.0 – non commercial – share alike – attribution page 40 table 3. sample demographics table 4. viewer correlation attitude towards ad a one-way analysis of variance (anova) was used to assess the relationships between categorical variables and attitudes towards the commercial. there were no significant differences in attitudes about the commercial for income level, age, education level, relationship status, or dog ownership. the anova for work status was significant, f(2) = 4.42, p = 0.013. post-hoc testing among categorical employment measures related to attitude toward the ad revealed that there was a significant difference between students’ attitudes (m = 3.02, sd = 0.81) and non-workers’ attitudes (m = 3.30, sd = .63), t = -2.45, p = 0.015, such that non-workers held more favorable attitudes towards the commercial than thomas mueller, twila wingrove, sidney murray pet behaviour science 2020, vol. 9, 34 44 doi:10.21071/pbs.vi9.12193 2020 vol. 09 34 44 www.petbehaviourscience.com creative common license 4.0 – non commercial – share alike – attribution page 41 discussion the results in this study support the findings of markovits and queen (2009), where those identifying at feminine were more attracted to the commercial than those who identified as masculine. the response to the commercial content could be associated with social identity or social proof, where those with similar values and virtues assimilate together. it is interesting to note that this commercial deals with the alleviation of loneliness, which would support the katz (2003) study stating that female attraction to pet rescue is based on feminine nurturing aspects, which are more compatible with dog rescue, as opposed to stereotypical male behavior which is purported to be aloof and disconnected from emotion. related to attraction to dogs, one might consider the work of amyx (2017) who stated cat lovers are individualistic, while dog lovers yearn for security and dependability. “somebody to love” in context provides the lonely man with both attributes and also promotes a greater connection with the community around the man and dog. the rescue dog – in an emotional sense – replaces a lost partner and stays at the man’s side, indicating a consistent, dependable nature. the practical nature of pet adoption may strongly affect perceptions of the commercial. non-workers, those who may have flexibility and free time at home, were most attracted to the pet rescue scenario. consideration could be based the margin of time, if resources are available to invest in the rescue animal. students and those in the workplace may have been affected by the advertisement, but the practicality of their personal situations may have been reflected in their responses. a bias exists, one that might explain the weak response from non-white respondents who viewed the commercial. the lead actor in the commercial is white. a photo displayed on the actor’s bed stand indicates there was a white, female partner who is no longer in relationship with the assumed heterosexual “lonely man.” the dog catcher depicted in the commercial is white, as are actors seen walking the street. one brown female is shown in the dog rescue shelter; however, she is placed in the background and is out of the field of focus. did students. there was also a significant difference between workers’ attitudes (m = 3.08, sd = .73) and non-workers’ attitudes (m = 3.30, p = 0.632), t = -2.68, p = 0.008, such that non-workers held more favorable attitudes towards the commercial than did workers.  the anova for ethnicity reached trend toward significance, f(1)  = 3.32, p = 0.069, albeit having a small minority group sample (n = 32). white participants reported more favorable attitudes (m = 3.18, sd = 0.69), while participants identifying as any ethnicity other than white held less favorable attitudes towards the commercial (m = 2.93, sd = 0.87).  finally, we tested for the combined utility of demographic variables in predicting attitudes scores. a multiple regression analysis revealed significant predictive power for the collection of independently significant or almost significant demographic variables of work status, ethnicity, and gender on attitudes towards the commercial, f(6) = 3.87, p = 0.001, with an r2 = 0.07. when all three demographic variables were included in the model, work status no longer predicted attitudes about the commercial: student versus nonworker, b = -0.42, p = 0.11; worker versus non-worker, b = -0.13, p = 0.62. on the other hand, both gender, b = -0.03, p = -0.01, and ethnicity were significant predictors, b = 0.29, p = 0.04. thomas mueller, twila wingrove, sidney murray pet behaviour science 2020, vol. 9, 34 44 doi:10.21071/pbs.vi9.12193 mueller, wingrove & murraypet behaviour science grooms and biddle (2018) suggest it is important to better understand the benefits of the canine-human relationship and to gain more knowledge related to that experiential bonding. this is a highly applicable advice related to context in pet rescue messaging. understanding how humans and animals build relationships, within the purview of gender and ethnicity, can open a new frontier for media facilitated animal rescue. creating association with rescue pets, while increasing support through gender identified community groups, is an advantage. coldwell banker and adopt-a-pet.com came together to present their plea to a national television audience, bolstered with local promotional efforts by area agents. the bond between like-believing individuals and their pets, was productive for both pet rescue and the real estate business. animal non-profit administrators, rescue shelter fundraisers and civic officials can apply findings from this research to develop targeted messaging, heighten interest in animal rescue, frame promotional campaigns and better connect with a diverse faction of animal-compassionate humans. marketing strategies specific to social identities and ethnicity can complement current pet rescue initiatives. a specialized, sensitive approach to these constituencies may help engender a positive attitude that can heighten intent to adopt rescue animals. identifying and targeting multiple personal identities and publics is essential if adoption groups and supporting sponsors wish to save the maximum number of rescue animals. embracing segmentation of ideologies, values, acquired knowledge and perceived relationships can work towards creating a bond between consumer, adoption organization and rescue animal (panoch & pearson, 2017). recognition of alternate publics and consumer segments will lead to a more realistic human depiction of the bond between adopter and rescue animal. the challenge to advertisers is which identifiers to feature. what is the engagement opportunity for building content around gender framed within a specific relational model? is there a possibility to target ethnicity and culture specific to geographic areas where adoption of abandoned animals is most critical? the context of home environment might also prove to be fertile for development of dog rescue commercials. working students, many in off campus housing, live busy lives that incorporate pet ownership. that lifestyle tests as significantly different from those who identified as non-working or stay at home parents. the differentiation may be framed based on optimum care for pets based on available time. advertising for pet rescue that acknowledges a differentiated clustering of potential adopters might adapt well to strategy in finding a common, healing message among discordant groups. society has become polarized through conflicted beliefs. consumers exist in social media “silos” that have become part of our new social identity. pfetsch (2018) states that although diversity of opinions are fundamental in pluralistic societies, politics are currently wrought with polarization, dissonance and eventually, disconnection from public spheres. public segmentation among religious groups may also heighten dissonance. lundy, adebayo and hayes (2018) suggest that for some, religion is perceived as a catalyst to peacebuilding. others view religion as exacerbating vengeance, conflict and hostility among social factions. rescue animals have the potential to bring a commonality of love and compassion to discordant identity groups. consider a commercial that presents a gender-feminine highly spiritual individual, finding common ground with a gender-masculine alt-right individual – through the conciliatory act of rescuing dogs through pet adoption. there are potential associations across political thomas mueller, twila wingrove, sidney murray pet behaviour science 2020, vol. 9, 34 44 doi:10.21071/pbs.vi9.12193 2020 vol. 09 34 44 www.petbehaviourscience.com views and gender identity, embraced through inclusive religious interpretation. while differences exist, the compassion and universal love for rescue animals can reign supreme. this study was conducted through a university classroom setting using convenience and judgmental sampling. the data collected is useful for testing in this pilot study; however, the demographics may not be representative of the general population. analysis of variance was utilized to identify significant differences among consumer segments. this study is quantitative in nature and does not disclose what prompts those disassociations. this would best be addressed through focus groups, inclusive of members stratified for gender identity, ethnicity and work status. it should be noted human identities are multi-dimensional. humanity is complex, as is each individual’s personal relationship with their pet. acknowledgments the authors wish to acknowledge students who participated in appalachian state university’s com 3928 research methods course as co-investigators in this study. they also wish to acknowledge statistical support and guidance from appalachian state university’s research design & analysis division in the office of research. the authors received no specific funding for this work. references amyx, d. 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(2017, march 27). coldwell banker’s adorable “somebody to love” ad extends commitment to pet adoption. retrieved january 3, 2018, from http://www.adweek.com/creativity/coldwell-bankers-adorable-somebody-tolove-ad-extends-commitment-to-pet-adoption/ thomas mueller, twila wingrove, sidney murray pet behaviour science 2020, vol. 9, 34 44 doi:10.21071/pbs.vi9.12193 2020 vol. 09 34 44 www.petbehaviourscience.com influence of dog presence on the tolerance and evaluation of aversive stimulation influence of dog presence on the tolerance and evaluation of aversive stimulation laura sodré galvão garcia*,1, isabela zaine1,2, camila domeniconi1 highlights • animals have been used for therapeutic purposes in various settings and can influence human well-being. • due to the history of domestication and cooperation with humans, the dog (canis familiaris) has the most potential to deliver benefits. pet behaviour science | 2016, vol.2, 16 – 23 doi: 10.21071/pbs.v0i2.4002 laura sodré galvão garcia*1, isabela zaine2, camila domeniconi1 1. department of psychology. universidade federal de são carlos 2. instituto de ciências matemáticas e de computação. universidade de são paulo departamento de psicologia fundação hermínio ometto – uniararas. paper research * email: lao_garcia@hotmail.com brazil. keywords: tolerance of aversive stimulation; humananimal relationship; dog; animal-assisted intervention • results indicate that that the presence of a dog mitigated the sensation of discomfort felt by the participants, enabling them to show more tolerance to the uncomfortable situation. • the present findings extend the current body of knowledge in the area of human-animal studies and have implications for animalassisted interventions. 1. introduction animals have been used for therapeutic purposes in hospitals, nursing homes and for people with various special needs since the 9th century (brodie and biley 1999). many animal species can influence human wellbeing. health benefits from interaction with pets may be explained by the social nature of humankind. however, due to the history of domestication and cooperation with humans, the dog has the most potential to deliver benefits (savalli and ades 2015). in fact, the first animal to be domesticated was the dog (de mello 2012; larson et al. 2012; axelsson et al. 2013). in therapeutic contexts, dogs have been used as distractors from situations that provoke emotional distress (brickel 1982). many studies have been conducted to understand people’s emotional responses to certain kinds of aversive sounds (zald and pardo page 16creative common license 4.0 – non commercial – share alike – attribution 2016 | vol. 2 | 16 23 abstract this study aimed to investigate the effects of the presence of a domestic dog in the evaluation and tolerance of auditory aversive stimulation. eighteen undergraduate college students participated. we analysed the latency of escape response from the aversive sounds and evaluation of the session through a semantic differential in three conditions: no distractors, a book of paintings as a distractor, and a dog as a distractor. latency of escape response was significantly higher in the presence of a dog and participants also evaluated the session more positively, suggesting that dogs can positively affect the perception of an aversive stimulation. http://www.petbehaviourscience.org/ 2001; cox 2007; cox 2008; neumann, waters and westbury, 2008; tajadura-jiménez et al. 2010). aversive auditory stimulation is known to stimulate the amygdala (zald and pardo 2001), a brain structure essentially related to emotional evaluation including the modulation of stress responses (oken et al. 2014). however, studies on human-dog relationships investigating aversive sounds as a stressful experience are missing. the mere presence of a dog during a task may affect physiological and/or behavioral indicators of wellbeing. house (1981) suggested that social support may reduce the perception of a stressful situation and may also tranquilize the neuroendocrine system so that individuals are less reactive to perceived stress. social support can come from different types of sources, such as a close friend, a spouse or even an animal (allen et al. 1991). in the last case, for example, kotrschal and ortbauer (2003) reported that the presence of a dog in a first grade classroom was associated to an enhancement of students’ behavior of paying attention to the teacher, as well as greater cooperation and social integration among peers. to understand the effects of the presence of domestic dogs on human health, researchers previously conducted correlational studies measuring heart beat and blood pressure (vormbrock and grossberg 1988; allen et al. 1991; eddy 1996; de mello 1999; allen et al. 2002; wolff and frishman 2004; wells 2005) as well as questionnaires and interviews (siegel 1990; hoffmann, et al. 2009; beetz et al. 2011). despite all reports about the positive effects regarding dog-animal interactions, it is still needed to produce researches with replicable methods and operationally described measures that may lead to behavior predictions. experimental analysis can more clearly demonstrate whether or not behavior can be attributed to environmental events. identifying the psychophysiological procedures that underlie in the interspecific relationships may provide a picture to the probable direct effect on human health by dogs (virues-ortega and buela-casal 2006). based on the results of previous literature, the purpose of this study was to verify the effects of a dog’s presence in the tolerance and evaluation of a situation in which the participants were exposed to aversive auditory stimuli in three different conditions. the removal of the headphone was defined as an escape response. tolerance to the aversive stimulation was inferred by the period of time participants endured the aversive stimulation. therefore, the longer participants remained with the headphones listening to the aversive sounds, the greater is the tolerance in relation to the sound. 2. methods participants eighteen undergraduate college students, 10 female and 8 male, aged 18 to 35 years (m = 25, sd = 2.37) participated. participants were self-recruited by informative flyers placed throughout the university campus. selection criteria were that participants did not fear dogs and did not have any hearing impairments. prior to the experiment, participants received information about the procedure, ethical concerns and signed an informed consent form agreeing to participate in the research. none of the participants withdrew from the procedure and they did not receive monetary compensation. moreover, our procedure was reviewed and approved by the committee of ethics in research with humans (protocol caae: 28071014.6.0000.5504) and by the committee of ethics in research with animals (protocol # ceua: 8077050415) of federal university of são carlos. materials and setting the experimental sessions were conducted individually in a room (2.5×2.5 m²) that contained a laptop, headphones and a futon for participants to sit on. the aversive stimuli, three low-pitched (1khz; 1.3khz; 1.6khz), continuous, monophonic sounds, were developed using the software audacity 2.0.5. halpern, blake and hillenbrand (1986) state that sounds composed by different low-frequency notes contribute to the perception of discomfort towards the auditory stimulation. participants were exposed only once to each one of the aversive sounds during the experiment to avoid habituation. since we used three different aversive auditory stimuli (1, 2 and 3) there were six possible sequences of sound presentation, from a to f, page 17 influence of dog presence on aversive stimulation creative common license 4.0 – non commercial – share alike – attribution garcia et al. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science throughout the procedure (a: 1-2-3; b: 1-3-2; c: 2-3-1; d: 2-1-3; e: 3-2-1; f: 3-1-2). the sequence of sounds was counterbalanced across participants and conditions, as shown in table 1. sounds were presented to participants via headphones connected to a laptop at 42 db, which is inside the limit established by the world health organization for sound levels heard via headphones – under 70 db (berglund, lindvall and schwela 1999). depending on the experimental session, distractor stimuli, either a dog or a book with paintings and text written in dutch were also present. we selected a book in an unfamiliar foreign language to avoid participants to engage in complex symbolic behavior, such as reading with comprehension. the dog was a one year-old male border collie previously trained to quietly lay down beside the participant and interact with him/her by eating dog treats. all sessions were videotaped for analysis of latency of removal of the headphones. a semantic differential scale was used to access participants’ evaluation of the aversive degree of the task. the semantic differential is a technique that permits measurement of the meaning of concepts, such as pictures, figures and, in our case, words (osgood, suci and tannenbaum 1957). participants had to assign likert-type scales ranging from -3 to +3 to 10 pairs of opposite adjectives: stressful / relaxing, bad / good, uncomfortable / comfortable, irritating / enjoyable, unpleasant / pleasant, restless / calm, exhausting / stimulating, boring / exciting, disturbing / peaceful and intolerable / tolerable. negative numbers correlate to a general negative evaluation of the task, positive numbers to positive evaluations, and zero to a neutral evaluation. for the first five pairs of adjectives, the numbers of the scale were organized from left to right starting from the number +3, that was the highest positive number. for the next five pairs, the scale started from left to right from the number -3, the highest negative number. this was done in order to avoid automatic responses to the numbers immediately below one another or side bias. procedure all participants were exposed to three different experimental conditions, control, book and dog, that were carried out once a day for three consecutive days. participants were randomly assigned to one of six possible sequences regarding the order of the experimental conditions, as presented in table 1. they were individually brought to the experimental room and instructed to sit on the futon. in the control condition, they were instructed to put on the headphones and to remove them and leave the room at the first sign of discomfort. in the book condition, the same instruction was given and the only change was that there was a book next to the futon that they could flip through while wearing the headphones. in the dog condition, they were instructed as in control and they were given the additional instruction that they could interact with the dog by playing with it or giving it treats while wearing the headphones. written instructions were given to the participants prior to session beginning in order to assure that instructions were consistently delivered across participants and sessions. in all conditions the sound was already being broadcasted via headphones, so that participants would hear it immediately upon wearing the headphones. we evaluated the tolerance of the aversive sounds by the latency until removing the headphones, measuring the amount of time that elapsed. after each condition, participants answered the semantic differential in order to assess their evaluation about the session. table 1. experimental conditions sequence assigned to participants. sound sequences were a: 1-2-3; b: 1-3-2; c: 2-3-1; d: 2-1-3; e: 3-2-1; f: 3-1-2. participants experimental sequence sound sequence p1 p2 p3 dog-book-control a b c p4 p5 p6 dog-control-book d e f p7 p8 p9 control-dog-book a b c p10 p11 p12 control-book-dog d e f p13 p14 p15 book-control-dog a b c p16 p17 p18 book-dog-control d e f page 18 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.2 | 16 23 data analysis tolerance to the aversive auditory stimuli was analyzed using the latency to escape from the aversive stimuli, which corresponded to the time elapsed between the participants’ positioning of the headphones and its removal. one-way between conditions analysis of variance (anova) for correlated samples was used to compare the average latencies across experimental conditions and participants’ scores on the semantic differential, followed by post-hoc tukey hsd tests to compare the two conditions with each other. pearson correlation tests were conducted between the latencies of escape response and scores on the semantic differential scale. all statistical analysis were two-tailed with a confidence level of 0.05. the data was analyzed using the statistical software spss version-20. 3. results participants showed a higher average of latency to remove the headphones in the dog condition (m = 24.4 min, sd = 10.6), followed by the book (m = 20.9 min, sd = 11) and control conditions (m = 15.1 min, sd = 6.3). this difference across conditions was statistically significant (one-way anova, f2,34 = 4.1, p <0.02). these results indicate that, in general, participants in the dog condition tolerated the aversive stimulation for longer periods of time. however, there was some variability. for example, latency from two participants significantly differed from the group mean: one participant from the book condition presented the highest latency score, enduring the aversive stimulation for 50 min; and one participant from the control group presented the lowest latency score, by immediately removing the headphones. the post-hoc tukey hsd test indicated that the latency to remove the headphones was significantly higher only when comparing dog vs. control (p <0.05) but no significant difference was found between dog vs. book and book vs. control conditions. this means that although participants usually endured the aversive stimulation for a longer period of time in the dog condition, the dog was not a significantly better distractor than the book. on the other hand, having the book of paintings as a distractor did not significantly alter the latency in the escape response when compared to the group that was not provided with any distractors. participants’ average latencies to remove the headphones across experimental conditions are shown in figure 1. figure 1. average latency of escape response in the control, book and dog experimental conditions. columns represent mean values. *tukey hsd test with p <0.05. error bars represent standard deviation. participants’ scores in the semantic differential also differed across the three experimental conditions. figure 2 shows the medians of the evaluations in each experimental condition. participants tended to give more positive evaluations in the dog condition (m = 1.2, sd = 1.3) and negative evaluations for the book and control conditions (m = 0.62, sd = 1.4 and m = 0.02, sd = 1.8, respectively). the difference across conditions was statistically significant (one-way anova, f2,358=71.2, p<0.001), with scores being significantly higher in the dog condition when compared to the book and control conditions and in the book condition compared to control (tukey hsd test, p <0.01). pearson correlation was used to calculate latency of escape response and the mean scores in the semantic differential to access the relationship between tolerance to the aversive sounds and evaluation of the sessions. there was a positive, though weak, correlation between the two variables (r =0.34 , n = 54 , p =0.009), meaning that participants who took longer to remove the headphones also tended to evaluate the sessions as more positive. these results are summarized in figure 3. page 19 influence of dog presence on aversive stimulation creative common license 4.0 – non commercial – share alike – attribution garcia et al. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science figure 2. median values of the semantic differential scale of the experimental sessions across different conditions (control, book and dog). the scatter plot was sectioned in four quadrants based on the medians of evaluation scores on the semantic differential and average latency of escape response. it can be observed that in the upper right quadrant, which represents the higher evaluations and latency of escape response, the predominant data points refer to the dog condition. this result indicate that although participants from the dog condition were exposed for more time to the aversive sounds, they also tended to evaluate the session using more positive describers of subjective private states. on the other hand, in the lower left quadrant, that represents the lowest evaluation scores and latency, the more prevalent data points represent scores from control condition, which indicate that when no distractors were available, participants tended to evaluate the session more negatively and endured the aversive stimulation for less time. figure 3. correlation between latency until escape response and mean scores in the semantic differential. tendency line represents pearson correlation, r = 0.34. dashed lines indicate the median values of the semantic differential score and latency to emit the escape response. 4. discussion our study aimed to investigate the impact of the presence of a dog in the tolerance and evaluation of aversive auditory stimulation. the results show that participants in the presence of a dog took significantly longer time to emit an escape response from the aversive sounds than in the control condition. moreover, they tended to evaluate the situation with the dog as more positive than when the distractor was a book of pictures or when there was no distractor available. also, there was a positive correlation between the latency of escape response from the aversive stimuli, experimental condition and evaluation of the session. in general, participants that rated the session as more positive also endured the aversive stimulation for a longer period of time, and the experimental condition that favoured higher latencies before the interruption of the aversive stimuli was the dog condition. our results corroborate other findings in the literature that point out that the short-term interactions with domesticated animals, dogs included, may decrease signs of physiological and behavioural distress (allen et al. 1991; odendaal 1999; hunt and chizkov 2014) and increase tolerance to aversive stimulation (kanfer and goldfoot 1966; peyron et al. 1999). as presented by savalli and ades (2015), health benefits from interaction with pets may be explained by page 20 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.2 | 16 23 the social nature of humankind. as such, humankind needs social acquaintances and, more than that, has the necessity of physical contact with living organisms. albeit the necessity of positive interactions exists in behavioral patterns of many living organisms, the need of attention becomes clearly identified only in advanced and social beings, as if it was a universal emotion, as stated by odendaal (2000). this may explain the fact that participants’ tolerance to the aversive stimuli did not significantly differ between control and book conditions, but differed between dog and control, suggesting that the dog was a stronger distractor, while the book of paintings did not significantly enhance participants’ tolerance to the aversive sounds. also, vormbrock and grossberg (1988) discussed that interactions with dogs might be less stressful than people due to classic conditioning because most people interact with pets in relaxed situations. thus the dog may be generally associated with activities that are positively reinforcing. to finalize, there are some limitations worth discussing. the first issue relates to the quality of the aversive stimuli here utilized. for ethical reasons we deliberately did not to use noxious or painful stimuli in our procedure. for definition, escape responses correspond to responses that lead to the interruption or termination of an ongoing aversive stimulation (catania, 1999). this way, escape responses are only emitted in the presence of aversive stimulation. we consider that any stimulation that a person behaves in order to escape is considered aversive (cooper et al. 2007), therefore, any noise that a person works to escape from can be considered aversive, which is the case of the sounds we used. also, there is evidence that continuous low-frequency sounds are usually perceived with discomfort (halpern et al. 1986). although, participants from the control and book conditions tended to use more negative describers, the adjective “boring” was the most frequent and salient one in subjects evaluation in the semantic differential scale. this might indicate that the sounds we used as aversive stimuli might have only been mildly aversive; therefore, participants were able to tolerate them for some time, with variation, in all experimental conditions. we consider that future research on this topic should use auditory aversive stimuli that have been empirically validated to control for these possible confounding variables. another possible limitation of our study was acoustic insulation. although the surroundings of the experimental room were relatively quiet and people were not allowed to transit in this area when the experimental sessions were ongoing, and the participants were wearing headphones, the experimental room was not soundproof. this way, it is possible that other sounds from the outside may have interfered at some level. a final concern refers to participants’ hearing. an inclusion criterion to our experiment was that participants did not have any hearing impairments, a condition that was self-stated. thus, future work might include audiometric screening of participants. 5. conclusion overall our study shows that the presence of a dog had a positive effect on the tolerance. here, we proposed an experimental study on the function of a dog as a distractor in the tolerance of aversive auditory stimulation, that are easily operationalized and replicable not only in experimental, but also in clinical and educational interventions. it would be interesting, though, to also include physiological measures of distress as complimentary data to gain a more comprehensive appraisal of the phenomenon. 6. acknowledgements isabela zaine was supported by a doctoral fellowship (fapesp, grant# 2011/06288-0). camila domeniconi was 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209–213, doi: 10.1002/smi.1057 wolff, a.i. and frishman, w.h. 2004. animal-assisted therapy in cardiovascular disease. seminars in integrative medicine 2: 4, 131-134. zald, d.h. and pardo, j.v. 2001. the neural correlates of aversive auditory stimulation. neuroimage 16: 746753, doi:10.1006/nimg.2002.1115 page 23 influence of dog presence on aversive stimulation creative common license 4.0 – non commercial – share alike – attribution garcia et al. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science influence of dog presence on the tolerance and evaluation of aversive stimulation predictors of proximity to others in colony housed shelter cats (felis silvestris catus) malini suchak, michael piombino, and kalina bracco highlights • cats showed high variability in time spent near conspecifics in the shelter • cats who were surrendered as strays were most likely to be classified as non-sociable pet behaviour science | 2016, vol.2, 24 33 doi: 10.21071/pbs.v0i2.5186 malini suchak, michael piombino, and kalina bracco animal behavior, ecology, and conservation canisius college, buffalo, ny paper research email: suchakm@canisius.edu united states keywords: cats, colony housing, felis catus, multi-cat household, proximity, shelters • none of the data collected upon intake significantly predicted which cats tended to be in proximity to each other 1. introduction from an evolutionary standpoint it is unknown exactly when or how the domestic cat (felis silvestris catus) evolved from a solitary ancestor to a species that, under the right circumstances, can and will form social bonds with each other (bradshaw et al. 2012). the beststudied examples of social behavior in cats come from free-ranging or feral cats who will form groups when there are ample resources as a result of provisioning (crowell-davis et al. 2004; bradshaw 2009; carfazzo and natoli 2009). these cats show a wide variety of affiliative behaviors including allogrooming, allorubbing, and sleeping in close proximity to each other. in general, agonistic behaviors are not frequently observed within feral cat colonies, although avoidance behavior such as turning the head away to avoid a direct stare may suggest that the cats are resolving page 24creative common license 4.0 – non commercial – share alike – attribution 2016 | vol. 2 | 24 33 abstract colony housing of cats allows shelters to maximize the number of cats housed in limited space. most research on colony-housed cats examines stress in relation to group size or enclosure size. while this is important for evaluating welfare, it is equally important to understand how cats are interacting socially in these colonies. we observed 259 adult cats housed in groups of two to eight individuals. scan samples were used to assess how frequently individual cats were in close proximity to other cats. these data were used to measure individual differences in sociability and patterns of proximity to certain partners. we used information about the past history of the cat, which was collected upon admission to the shelter to identify predictors of time spent in proximity. there was a high degree of inter-individual variability in sociability. strays tended to spend less time in proximity to other cats, and this effect was most pronounced in females. however, none of the information collected upon admission predicted patterns of proximity to certain partners, or which cats spent time in association with each other. future studies should explore the implications of differences in sociability by associating observations of social behavior and stress behaviors. http://www.petbehaviourscience.org/ conflicts through subtle signals rather than overt fighting (dards 1983). given their large home ranges, avoidance may be more important than overt aggression for cats. the potential use of large spaces by free-ranging cats to avoid conflict has led many to question how they adapt to social living in the home, where the space per cat is typically of an order of magnitude less than outdoors. cats may form individual territories or home ranges within the house and frequently spend time out of sight of each other (bernstein and strack 1996; barry and crowell-davis 1999). they even seem to “time share” favored spots, each occupying the spot at a different time of day (bernstein and strack 1996). familiarity and relatedness are important factors in whether or not household cats spend time together, with related individuals spending the most time together (bradshaw and hall 1999; curtis et al. 2003; crowell-davis et al. 2004). overall, the rates of aggression observed in the household are low and are most often negatively correlated with familiarity (bernstein and strack 1996; barry and crowell-davis 1999; bradshaw and hall 1999). the role of familiarity on aggression and affiliation has significant implications for shelters that wish to house cats in groups. group housing may have several benefits. depending on the amount of space and provisioning of environmental enrichment, cats that live in a larger enclosure may have more hiding places and perches, which are important for reducing stress (casey and bradshaw 2007; kry and casey 2007; ellis 2009). furthermore, cats housed in communal housing are adopted just as quickly as cats housed in enriched, single cages, and more quickly than those housed in basic, single cages (gourkow and fraser 2006). however, given that the space per cat is significantly less than they would have in the household, there may also be costs to living in a group without the opportunity to avoid one another by leaving (ottaway and hawkins 2003). most of the work done on group housing and cats has focused on the stress levels of cats (finka et al. 2014). while stress is undoubtedly important in assessing the welfare of shelter cats, little attention has been given to whether or not the cats are engaging in positive social interactions or avoiding each other. since socialization appears to occur during a critical period of 3-7 weeks of age (landsberg 1996), it is unlikely a shelter will have knowledge of the socialization of an adult cat, particularly for those individuals who are surrendered with unknown histories or from single-cat households. coming from a multi-cat household may ease the adjustment of a cat even in single caging, as they are used to having other cats around (broadley, mccobb and slater 2014). the goal of the current study was to identify predictors of time spent in proximity to conspecifics from the information collected upon admission to the shelter. many previous studies have established that proximity is a reliable measure of affiliation between cats (macdonald and app 1978; van den bos 1994; barry and crowell-davis 1999; curtis et al. 2003; crowelldavis et al. 2004) and many other species (dunbar and shultz 2010; silk et al. 2013,). based on the literature, we hypothesized that cats who were surrendered together from the same household were more likely spend time in proximity than cats who were previously strangers. furthermore, we hypothesized that if the individual history of the cat, such as their previous living situation (owned versus free-ranging), or the number of other cats they lived with reflected social experience, then cats that had indicators of previous social experience may spend more time in proximity to others. alternatively, if the cats were primarily seeking to avoid each other in the shelter colony (sensu ottaway and hawkins 2003), we predicted that a higher density of cats in the room (number of cats per m2 of space) would lead to increased proximity between cats. 2. methods participants and setting. participants were 259 adult cats (128 males and 131 females) of various breeds, including mixed breeds, housed at the spca serving erie county. group sizes ranged from two to eight cats, with a median of three cats per group. the staff at the spca decided group composition upon admission. there were no strict criteria for inclusion in a group, although cats that had contagious medical issues or were surrendered for a page 25 predictors of proximity in shelter cats creative common license 4.0 – non commercial – share alike – attribution suchak et al. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science history of aggression towards other pets were excluded from the groups. upon introduction, a staff member or volunteer observed the group for approximately an hour in case of severe aggression. in total, we observed 87 groups of cats in this study. among the groups we observed there were single-sex (n = 32) and mixed sex groups (n = 55), as well as groups of individuals that were brought in together (n = 41) and groups where individuals were introduced at the shelter (n = 46). the groups were built on an all-in, all out policy, that is, individuals were placed together and no new individuals were added until all of the individuals were adopted out. thus, there were cats leaving the groups, but not entering the groups during the time of the study. the average length of stay (los) in the shelter was 39 days (range: 1-205) for our participants. this measure, called shelter los throughout, represents the total amount of time from intake to the date of observation, including time when the cat was not available for adoption or present in the colony housing. on average, cats only spent 15 days in the colony setting (range 1-85, but note that only 6 cats spent more than 39 days in colony housing). this measure is called colony los throughout. since groups were created using an all-in, all-out policy, this represents the length of time each group was together, on average. the cats were housed in one of four colony rooms, ranging in size from 3.06m2 to 5.41m2. each room contained 1-2 litterboxes, several beds and towels, and typically contained a karunda bed ®, 1-2 plastic milk crates, and a long shelf approximately 1.19m high. the two larger rooms also contained a mid-level shelf (approximately 0.6 m high). food and water were provisioned by the shelter staff and at no time were participants food or water deprived as part of this study. this study was approved by the spca serving erie county prior to the commencement of data collection. this study did not require approval of the canisius college iacuc because it was observational in nature. background data on the cats. using the spca’s petpoint database, we gathered background information on each of our participants that was collected upon admission to the shelter. this included the cause for relinquishment (owner surrendered (n=192), stray (n=37), or seized (n=30)), number of cats in the intake (number admitted together, m=6.6, range 1-50), and names of cats that came in together. for the purposes of this study, any individuals that were brought in together were considered “familiar” and any individuals that were not brought in together were considered “strangers.” based on the intake data we were unable to determine relatedness or length of time spent together for familiar individuals. we also recorded demographic information including sex, age, and spay/neuter status. procedures. observations were collected from january 2014december 2015 (92.6 hours of observation). each group was observed for 10 minutes, twice per day, 2 to 4 times per week. because we could not control when cats were adopted, not all cats were observed for the same amount of time, but all cats were observed for at least two sessions. the average number of scans each cat was observed was 71.3 (range: 22-251). all observations occurred between 10:00h and 16:30h, during the normal operating hours at the shelter, and were balanced across time of day. observations were suspended during husbandry procedures, or if shelter staff or visitors entered the room. thus, observations were limited to periods when just the cats were present in the room without any human interference. during each 10-minute observation period, the observer stood outside the clear glass door of the colony room. scan samples were taken at 1-minute intervals to record the proximity of each cat to all others. two short, separate observation periods per day ensured that we were not overestimating the preferences of 2 cats who happened to be sleeping near each other at any given time. each cat was scored as being in contact with another cat, within one body length but not touching, or greater than one body length away from each other cat. only cats who were resting, sleeping, or grooming with another cat were scored as in proximity. thus, cats who were transiently passing by other cats were not counted as in proximity. as per previous research (macdonald and app 1978; van den bos 1994; barry and crowell-davis 1999; page 26 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.2 | 24 33 curtis et al. 2003; crowell-davis et al. 2004) time spent in proximity can be used a proxy for tolerance towards another. although previous research has used <1m as a measure of proximity, we chose one body length as a more conservative measure given that the size of the room would automatically put most cats within one meter, rendering this measure useless in determining tolerance. body length is a frequently used measure of proximity in other species (i.e., primates: perry 1996; cetaceans: conner et al. 2006) and is defined as the length from nose to tailbase of an adult individual. we also recorded all occurrences of any aggressive interactions including swat, hiss, growl, and chase. inter-rater reliability was excellent on all measures (97.78% agreement between raters), and was collected by two live scorers present in front of the colony enclosure for a random subset of sessions. cats were not videotaped due to blind spots in the room (e.g., hiding places). analysis each cats’ individual propensity to spend time near conspecifics (regardless of partner) was calculated using as the percentage of scans in which an individual was in proximity (defined as a body length or closer) to another cat. because the data from each scan are not independent from each other (a cat sleeping in proximity during one scan is likely to be in proximity the next), data were collapsed across the day. due to the large number of zeros (cats who were never within a body length of another cat) in the data, we collapsed this measure into a binary sociability index, designating each cat as either sociable or non-sociable (1/0). cats who were never within a body length or closer of another cat were scored as non-sociable (0), and cats who were, at some point, within a body length or closer of another cat were scored as sociable. we used the background data collected from the petpoint database to see if any characteristics of the cats predicted whether or not an individual might be classified as sociable. to control for repeated sampling of individuals, we ran a generalized linear mixed model (glmm), using the information criterion (ic) method for selecting the best model (bolker et al 2009). the ic method uses model selection to compare fits of models, estimating their predictive power on the dependent variable. since the current study was interested in which factors were predictive of proximity to other cats in the shelter, the ic method was used to identify these factors. the purpose of running multiple models testing different combinations of the fixed effects is to determine which fixed effect or combination of effects is the best predictor of sociability. we determined the model with the most explanatory power by comparing the akaike information criteria (aic) values for all of the possible models. we then tested the best fit model against a full model with all of the fixed effects and a null model with only the random effects. all analyses were run using r statistical software (r development core team, 2014) using the glmer function of the lme4 package (bates, maechler, and bolker 2012). after obtaining aic from all of the models, we used the anova function to determine which model had the most explanatory power. the significant interaction was explored using the ggplot function. the binary sociability index (1/0) was the dependent variable. individual, date of observation, and observer were entered as random effects. fixed effects tested included the density of cats in the room (cats/m2), number of cats in the intake, source of intake (categorical: owner surrendered, stray, or seized), spay/neuter status (1/0), age, sex, length of stay at the shelter and length of stay in the colony housing. since data were aggregated by day, we calculated the length of stay measures specific to each observation day; that is, for shelter los, we calculated the difference (in days) between the date the animal came into the shelter and the date of observation, for colony los it was the difference between when the cat came into the colony and the date of observation. we tested density, number in intake, and source of intake individually and also the 2-way interaction of these effects with demographic information such as spay/neuter status, age, sex and the two los measures. since shelter los and colony los were highly correlated (r = 0.507, p < 0.001), we did not include those two measures in the same model together, but rather ran separate models for each of them. see table 1 for a summary of models. due to frequent changes in group composition, most typical measures of patterns of proximity (e.g., affinity matrices, social network analysis, etc.), are not page 27 predictors of proximity in shelter cats creative common license 4.0 – non commercial – share alike – attribution suchak et al. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science appropriate for this setting. we calculated patterns of proximity using the percentage of scans in which two individuals were in proximity (a body length or closer) to each other. there were also a large number of zeros for this measure (e.g., pairs of cats who were never in proximity to each other), so we again created a binary variable, designating each pair as either in proximity or not (1/0). we again used a glmm to determine which predictors had the most explanatory value in proximity, our dependent variable. random effects were dyad, date and observer, to control for repeated sampling of the same individuals across multiple days. fixed effects tested included whether the cats were familiar or strangers, sex composition of the dyad (e.g., m-m, m-f, page 28 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.2 | 24 33 table 1. summary of models tested for predictors of proximity. a model including source of surrender, sex, and the interaction between source and sex was the best fit model. and f-f), and days together in the colony setting. among dyads who were at some point in proximity (dyads scored as 1 above, n = 142) we ran a follow-up analysis to see if there was a difference in dyads who were in contact (e.g., touching) vs. those who were within a body length, but not in contact. the dependent variable was contact, the random effects, fixed effects, and models were the same as above. we were unable to analyze aggression as it occurred too rarely (only on 3 occasions). 3. results sociability. there was extremely high inter-individual variability in sociability (as measured by percent of scans in proximity to another individual; figure 1). one hundred ten cats (42.47%) were never seen in proximity to another individual, the remaining ranged from 4.34% to 100% percent of scans in proximity to another cat (median = 4.50%). only 40 cats (15.4%) were recorded in proximity to other cats more than 50% of the time. the results of the glmm revealed that a model including the reason for surrender and the sex of the cats had the most explanatory power in whether or not a cat was categorized as sociable (aic = 921.81, χ2 = 20.74, χ2 df= 5, p < 0.001; table 1). the intercept was not significantly contributing to the fit of the model. there was a main effect of source (z = -3.53, p < 0.001) and a significant interaction with sex (z = 2.59, p = 0.01). a closer look at the data revealed that both males and females that were surrendered to the shelter as strays were less likely to be classified as social (figure 2). however, this difference was much more pronounced for females than males, with female strays only having a 0.10 probability of being classified as social. the results of the second glmm on patterns of proximity revealed that the model including familiarity had the most explanatory power (aic = 929.24, χ2 = 2.5, χ2df = 1, p = 0.11; table 2), however it was not significantly better at explaining the variance than either the null or full model. a closer look at the best fit model revealed that the intercept was significant (z = -3.54, p <0.001), but familiarity was not (z = 1.61, p = 0.11). thus, none of the fixed effects tested in this study significantly predicted time spent in proximity to a specific partner. when the analysis was limited to only cats that were in proximity, and compared those in contact versus those within a body length, but not in contact, the null model had the most explanatory power (aic = 287.67, table 3). again, none of the fixed effects predicted time spent in contact over general proximity, there was only a trend towards significance for the random effects (z = -1.92, p= 0.052). page 29 predictors of proximity in shelter cats creative common license 4.0 – non commercial – share alike – attribution suchak et al. figure 1. variation in proximity to conspecifics. there was extremely high inter-individual variability among cats regarding the amount of time spent in proximity to conspecifics. the histogram shows the number of cats by percent scans in proximity, grouped into blocks of 10%. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science model model df aic deviance null (random effects only) 4 929.75 921.75 full (all fixed effects) 8 934.21 918.58 familiarity (best fit) 5 929.24 919.24 other models tested sex composition 6 932.90 920.90 time together 5 931.56 921.56 familiarity * sex composition 9 933.91 915.91 familiarity * time together 7 933.13 919.13 sex composition * time together 9 936.58 918.58 table 2. summary of models tested for patterns of proximity. a model including whether the cats were familiar (came from the same intake), was the best fit model, but did not explain the variance better than the null and full models. model model df aic deviance null (random effects only) 4 287.67 279.67 full (all fixed effects) 8 295.24 279.24 other models tested familiarity 5 289.35 279.35 sex composition 6 291.56 279.59 time together 5 295.24 279.66 known * sex composition 9 291.72 273.72 known * time together 7 291.73 277.73 sex composition * time together 9 295.37 277.37 table 3. a summary of the models tested for predictors of which days might be in contact versus simply within a body length, but not touching. the null model had the most explanatory power. 4. discussion in this study we used proximity as a behavioral measure to assess how cats in colony housing interact with each other. overall, the cats varied highly on propensity to be sociable with one another. approximately 42% of the cats were never in proximity to another cat during the period of observation. there might be several reasons why cats may try to maintain some distance in the colony room. first, cats may wish to avoid social contact with other cats. the use of body length as a measure in other work is often chosen because it is close enough between two individuals that there is potential for contact (i.e., perry 1996). keeping a further distance may allow individuals more of an opportunity to avoid contact. although nearly half of the cats observed in this study were more than a body length away, if they were seeking to avoid contact, we cannot determine from the current data if there was sufficient space for them to do so. an alternative explanation is that non-social cats were avoiding other cats due to stress. stress can lead to suppression of behavior, and while this is typically applied to page 30 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.2 | 24 33 figure 2. cats surrendered as strays are less likely to be classified as social. this effect was more pronounced in females (circles) than males (triangles). error bars represent interquartile range (first through third quartile). maintenance behavior, it could impact social behavior as well (rehnberg, robert, watson and peters 2015; stella, croney and buffington 2013). a behavioral or physiological measure of stress could shed light on how frequently highly stressed cats are in proximity to others. on the other hand, a large number of cats spent time in proximity to each other, some of them frequently so. as determined by the glmm, the best predictor of sociability was a model including sex and source of intake (owner surrendered, stray, or seized). overall, cats that were surrendered as strays were less likely to be categorized as sociable. it is important to note that this cannot merely be explained by number in intake or number in the room, as both of these factors were accounted for in the analysis and source alone was the best predictor. the observed interaction between source and sex is puzzling, as female strays were less likely to be categorized as sociable than male strays. many studies have found that female feral cats tend to have much stronger social bonds than males (macdonald and apps 1978; crowell-davis et al. 2004; bradshaw 2009; carfazzo and natoli 2009; houpt 2011) and have smaller home ranges, suggesting greater tolerance than males (dards 1983). however, these cats were classified by the shelter as “stray” rather than feral, since they were socialized enough with humans to be placed on the adoption floor. far more is known about the social interactions of owned cats (i.e. bernstein and strack 1996; barry and crowell-davis 1999) and feral cats (i.e. carfazzo and natoli 2009), than free-ranging socialized cats. previous work has shown owner surrendered cats tend to be more stressed than strays (dybdall, strasser and katz 2007) and that male cats tend to be more stressed than females (rehnberg et al 2015). thus, our findings demonstrate cats less likely to be stressed (females, strays) are also less likely to be in proximity to other cats. although above we note that stress may lead to the suppression of social behavior, theoretically it might also lead to increased proximity. for example, if an individual is very inactive, they may not move away from another individual, even if they prefer to be alone. this would, on the surface, look like proximity, but have nothing to do with tolerance or a preference to be near others. second, the cats observed in this study are on the adoption floor. members of the public frequently enter the rooms to interact with the cats. there may be a “safety in numbers” effect, whereby proximity to other cats lessens the chance of someone directly approaching a particular cat for interaction. more detailed knowledge of how cats end up in proximity (e.g., who approaches whom?), measures of stress, and observations of the distance between cats when people are in the room may help determine how the setting is impacting their social spacing. we also looked at patterns of proximity to see if there were particular individuals who tended to be near each other. such a measure has implications for the cats’ well being if not adopted together or moved independently while at the shelter, and also might help shelters decide which cats to place in rooms together. however, none of the data collected upon intake predicted patterns of proximity, and our hypothesis was not supported. this is in contrast to previous reports where familiarity predicted proximity; however, it is important to note that those were studies of homed cats whose histories were known in much more detail (e.g., bradshaw and hall 1999; curtis et al. 2003). we did not know how long two cats had been together or kin relationships, two factors, which in these previous studies of homed cats have been implicated in affiliation patterns. behavioral signs that two individuals are closely bonded, such as allogrooming, or signs of distress when separated may be informative to shelter staff in addition to time spent in contact or proximity. however, it is important to consider that stress may also suppress these behaviors as well. in addition to collecting stress data, longer data collection periods may also shed light on patterns of proximity better than the short, multiple data collection periods used in this study. we chose to use two short sessions per day so as to capture, but not overrepresent proximity between two cats who might be sleeping near each other. this created a significant limitation as doing two short sessions may have missed some opportunities for particular pairs to be near each other. we also rarely observed activity, limiting our opportunity to observe aggression and affiliative behavior (such as approaches with a tail up, nose to page 31 predictors of proximity in shelter cats creative common license 4.0 – non commercial – share alike – attribution suchak et al. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science nose sniffing), which occur randomly and infrequently throughout the day. furthermore, because our observations were live, we were limited to only the times in which the shelter was open to the public. given the potential role of stress in social behavior, collecting data when the shelter is not open could be highly useful in understanding the patterns observed in our study. 5. conclusion understanding the group dynamics of cats living in shelter colonies can help shelter employees make informed decisions about how to place cats upon admission. our data contribute to this growing body of knowledge, specifically by suggesting that strays are less likely to be in proximity to other cats in colony housing. however, given the puzzling sex difference observed and limited observation time, our results should be interpreted with some caution. future research should investigate the interaction between stress and social behavior in colony housed cats. 6. acknowledgements we would like to thank the spca serving erie country for allowing us to collect data on their colony housing and julia watzek for statistical advice. this research was funded in part by a grant from the al and noura gress foundation. 7. references barry, k.j., and crowell-davis, s.l. 1999. gender differences in the social behavior of the neutered indoor-only domestic cat. applied animal behaviour science 64: 193-211. bates, d., maechler, m., and bolker, b. 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lybica f. catus l.): a study of dominance in a group of female laboratory cats. ethology 98 14-37. page 33 predictors of proximity in shelter cats creative common license 4.0 – non commercial – share alike – attribution suchak et al. http://www.r-project.org/ http://www.r-project.org/ https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science integrating animals in the classroom: the attitudes and experiences of australian school teachers toward animal-assisted interventions for children with autism spectrum disorder. bradley p smith and ashley a dale1 pet behaviour science | 2016, vol. 1, 13 – 22 bradley p smith and ashley a dale 1. central queensland university paper research email: b.p.smith@cqu.edu.au australia keywords: animals; companion animals; animal-assisted activities; autism spectrum disorder; special education; school; teacher attitudes. highlights providing children with the opportunity to interact with animals in classroom settings has many potential benefits, especially for individuals with autism spectrum disorder. yet, our understanding of the experiences and attitudes of school teachers towards integrating animals in the classroom is limited. page 13creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 13 22 abstract the introduction of animals into school classrooms has been posited as a beneficial intervention for individuals with autism spectrum disorder (asd). whilst evidence that animal-assisted interventions or activities can positively influence classroom behaviour and learning outcomes is emerging, little is known about the experiences and attitudes of those who implement it. we presented a series of open and closeended questions via an online survey to australian school teachers working with students on the autistic spectrum. whether teachers had experienced companion animals in the classroom or not, companion animals were believed to provide a means for improving social skills and engagement within the classroom, as well as decreasing stress, anxiety, and the occurrence of problematic behaviours. yet, despite an overall positive attitude, and 68% having had animals or pets in their classroom, only 16% of respondents had experience with ‘formal’ animal-assisted interventions. explanations for why both formal and informal animal-assisted interventions were either not being adopted, or was not currently being considered, included a lack of knowledge, lack of support and resources, reactions of the student in relation to allergies and behaviour, and issues relating to animal welfare. it was also acknowledged that the evidence-base for animal-assisted interventions for students with asd is currently lacking, and that such interventions were not suitable for all students, or all classroom situations. moving forward, it is important that the inclusion of companion animals and more formal based animal intervention programs in classrooms be adequately designed and evaluated, because implementing or promoting time consuming and financially costly strategies without the evidence is problematic. http://www.petbehaviourscience.org/ this study begins the process of uncovering some of the benefits and barriers relating to the integration of animals in australian schools. the inclusion of animals in classrooms, particularly dedicated animal-assisted therapies, needs to be adequately evaluated, as introducing time consuming and financially costly interventions without adequate evidence is problematic. 1. introduction the physical, social and emotional benefits that can be achieved through human-animal interactions has led to the rise of animal-assisted interventions as a treatment option for a variety of clinically diagnosed conditions (nimer and lundahl 2007). this includes neurodevelopmental disorders such as autism spectrum disorder (asd) (grandin et al. 2010; o’haire 2013). asd is a chronic disorder that emerges throughout childhood. it is characterised by significant deficits in social interaction and communication, and the presence of restricted and repetitive interests (american psychiatric association 2013). these deficits often lead to on going negative consequences for the individual, family (smyth and slevin 2010), and the economy (roth 2013). a sharp rise over the last decade of children diagnosed with asd (roth 2013) has signalled a need for effective interventions, particularly those that are economically viable (lord et al. 2005), and can be conducted in school-based environments (anderson and olson 2006; berry et al. 2013). one viable treatment option available for teachers, school counsellors and psychologists in educational settings involves the promotion of animal contact (kaufmann 1997; chandler 2001). the term ‘animalassisted interventions’ has been used to describe the formal (animal-assisted therapy) as well as the less formal (animal-assisted activities) treatment options. animal-assisted therapy involves the deliberate and purposeful inclusion of an animal to help participants meet specific treatment goals that may be best addressed or facilitated with the assistance of the animal. however animal-assisted activities encourages activities with animals to provide opportunities for motivational, educational, recreational, and/or therapeutic benefits without the presence of specified treatment goals or objectives. the latter does not have to be implemented by trained personnel, and session content is spontaneous (nimer and lundahl 2007; fine 2010; o’haire et al. 2014). numerous studies have highlighted the benefits of animal-assisted interventions for school aged children, particularly children on the autistic spectrum (hergovich et. al. 2002; kotrschal and ortbauer 2003; jalongo et al. 2004; anderson and olson 2006; friesen and delisle 2007; daly and suggs 2010; grandgeorge et. al. 2012; o’haire et al. 2013; o’haire, et al. 2014). the ability of an animal to change the dynamic of the classroom to be more positive and engaging has been recognised and harnessed into the creation of specific animal-assisted programs, such as ones focusing on literacy (jalongo 2005). domestic dogs, with which children with asd can form strong bonds (carlisle 2015), have been successfully used in classrooms to support student learning through modelling trust and acceptance, and have been linked to an increase in positive attitudes towards other students, learning, and attendance (anderson and olson 2006; friesen and delisle 2007; jalongo et al. 2004; beetz 2013). animal-assisted interventions may also serve as a conduit for pro-social behaviours, such as social inclusion, play, communication and bonding opportunities for asd students (friesen and delisle 2007; o’haire et al. 2013). in one study for example, typically developing peers were ten times more likely to interact with a fellow student who had a disability if they were in the company of a dog (jalongo et al. 2004). several evaluations of animal-assisted programs for children with asd have reported increases in overall social and emotional behaviours such as verbal, gestural and visual communication towards the animal, teachers’ and peers, and decreased self-absorption and self-stimulatory play (kršková et al. 2010; o’haire et al. 2013; silva, et al. 2011; carlisle 2015). teachers regularly report positive changes in students when animals have been involved in the classroom, such as sustained emotional benefits to animal and human-directed empathy, increased social interactions, reduced aggressive behaviours and a decrease in hyperactivity, all of which are domains where students with asd require support (ascione and weber 1996; hergovich et. al. 2002; daly and suggs 2010). page 14 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 13 22 although there is a great deal of anecdotal evidence that animals can lead to beneficial outcomes for certain individuals with asd, there are few scientifically rigorous studies evaluating the use of assistance animals in classroom settings (see nimer and lundahl 2007 and o’haire 2013 for reviews). the evaluations that exist suggest that the use of animals in classrooms (and elsewhere), although promising, suffer from methodological issues (e.g. single-subject experimental designs), only show small to moderate effects in relation to improving outcomes, and have yet to be shown to promote long term and meaningful change (nimer and lundahl 2007; o’haire 2013; kasari and smith 2013). part of the issue with conducting such evaluations is the breadth of animal-related interventions available, the contexts and disciplines in which the use of animals in classrooms have been implemented, and the teaching strategies used (likely a reflection of inadequate training, support, and intervention manuals available). in addition to a paucity of scientific evidence, several practical barriers to introducing animals in classrooms have been identified (hergovich et. al. 2002; kotrschal and ortbauer 2003; prothmann et al. 2009). a study of teachers in the united states highlighted several factors preventing teachers from having classroom pets including policy and legal concerns; a dislike of animals; a lack of adequate space for the animal; and the potential of an increased workload (rud and beck 2003). ladarola et al. (2015) found several challenges in introducing new interventions into schools for children with asd. these included tension between stakeholders (i.e., between staff and administration, staff and parents, special education and general education teachers), lack of training, and a lack of culture of accepting difference. it is likely, but uncertain whether these perceptions and challenges exist in the australian school context. teachers’ play a pivotal role in the implementation and success of classroom interventions. understanding the association between teacher attitudes, experiences, and their likelihood of integrating animals in classrooms therefore, is paramount to animal-assisted interventions becoming more widely practised. it also increases the opportunity for such interventions to be appropriately evaluated. this study begins the process of uncovering some of the obstacles that may prevent the successful integrating of companion animals and animal-assisted interventions in australian schools from the perspective of those that implement it. 2. methods participants a total of 73 current australian school teachers working specifically with students diagnosed with autism spectrum disorder participated in the online survey (67 female, 5 male). just over half were aged between 25 – 34 years (53.0%), with 15.8% between 45 and 54, 10.5% between 55 and 64 years, 13.2% between 35-44 years, 6.6% between 18-24, and 1.3% between 6574 years of age. the majority of the sample had positive inclinations towards animals, with 90.4% identifying as ‘an animal person’, and animal ownership among the sample (74.0%) slightly above the national average (australian companion animal council, 2010). the total amount of teaching experience varied across the sample, with 65.8% having between 1 and 10 years experience, 13.7% between 11 and 20 years, and 17.9% with 21 or more years experience. the majority of the teachers worked in state/government schools (78.1%), and identified the schools as residing within either an urban (68.5%) or rural setting (31.5%). participants taught across all year levels, with 32.9% working in a primary school, 24.7% in a secondary school, and 9.6% in a joint primary/secondary (prep to year 12) school. about one third of teachers (32.9%) worked specifically within a special education context. materials the research team designed an online questionnaire based on the literature on animal-assisted interventions in general. the survey included a maximum of 58 question items (41 closed and 14 open-ended questions). the exact number of questions answered was determined by responses to specific items. questions required responses to a mixture of yes/no questions, list of options, likert-type scales, and openended questions. the questionnaire covered topics relating to the participant (e.g. gender, age, education, page 15 attitudes of australian school teachers towards animals in the classroom creative common license 4.0 – non commercial – share alike – attribution smith, d.p. and dale, a.a. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science training specialisation, current position, number of years teaching); the school where they were employed at the time of the survey (e.g. number of students, location, percentage of students that own pets, the degree to which animals play in the lives of the children); their attitudes towards animals (e.g. current and past animal ownership, whether they identify as an animal person, negative experiences with animals); their experience and knowledge of animal-assisted therapies (e.g. whether they had read any information on animal-assisted therapies and what prompted the search, what animals they believe would be effective in the classroom, training with animal-related therapies, whether the school has active animal programs, the details of such programs including what sessions or activities animals were used for, and reasons for involving animals); potential barriers that exist preventing them from having used or including animals in the classroom; and resources that might be needed in order to develop an animal-related program. open ended questions included, for example; “what is the first thing that comes to mind when you hear the phrase animal-assisted therapy?“; “in 150 words or less, please provide a brief example of how animalassisted therapy was successful [or not] in achieving your initial goals”; “in 150 words or less, please provide some insight into your likelihood of continuing, or not continuing, to use animal-assisted therapy”; “please explain or list some of the reasons why you have not had a pet in the classroom”; and "do you have any other comments relating to animalassisted therapy and children with autism spectrum disorder that you would like to share?". questionnaire is available from the author by request. procedure australian teachers with experience teaching children with autism spectrum disorder were invited to partake in an anonymous survey through various online interest groups that encouraged teachers to communicate and share knowledge and best practices. members who chose to participate were directed to an online survey and provided further detailed information about the study and a consent form. the survey was available between july and october 2014, and took approximately 20 minutes to complete. the project was approved by the central queensland university human research ethics committee (h14/05-094). analyses closed-ended response items were analysed using descriptive statistics. a thematic analytical approach (braun and clarke, 2006) was utilized to identify the main themes within the data corpus. thematic analysis is a rigorous method of discourse analysis useful for analysing patterns (themes) within a data corpus. the first stage of analysis requires data familiarisation, and therefore involved reading over the qualitative responses. in the second stage, the data were searched, asking three major questions: (1) what are the motivators for integrating companion animals into the classroom; (2) what are the perceived benefits of integrating companion animals into the classroom; (3) what are the perceived barriers to the integration of animals into the classroom? extracts from participants’ responses were collated under each of these three questions. these extracts were discussed within the research team to establish agreement that they were coherent with the themes and representative of the patterns across different participants’ responses (green et al. 2007). 3. results experience and knowledge of animal-assisted classroom interventions the majority of teachers had had animals or pets in their classroom (68%), with a total of 14 teachers (18% of respondents) reported having directly experienced formal animal-assisted therapy in the classroom. only 5% of all teachers were currently working in a school that had an active animal-related program. the majority of teachers who had incorporated animals worked in special education schools (50%), were situated within a city/metro location (75%), and were government based/public (66.6%). the experience of teachers who had engaged with animal-assisted interventions varied, with years of teaching ranging between 5 to 44 years of age. teachers were mostly between 55-64 years (66.6%), followed by those between 25-34 years (33.3%). for teachers who had not been directly involved with animals in the classroom, page 16 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 13 22 an overwhelming number (96.7%) expressed interest in pursuing it in the future. when asked to rank a provided list of 10 animals by their perceived effectiveness for classroom based interventions, domestic dogs were ranked as number 1 by 60/75 (80%; mean ranking 1.77), followed by cats as number 2 (38%; mean ranking 4.2), and rabbits as number 3 (31%; mean ranking 3.97). animals used in existing animal-therapy programs primarily involved the use of domestic dogs (n=3), but the list of species extended to horses, rabbits, guinea pigs, chickens, and farm animals (with each species/group being identified by one respondent) a quarter (26%) of teachers surveyed had read literature relating to animal-assisted interventions. when prompted for the source of the information, participants mentioned academic journal articles, followed by newspaper articles, set readings as part of a university course, and the internet. when asked what encouraged them to research animal-assisted interventions, few had actively sought out the material, but had heard about it from colleagues, reading about it whilst studying at university, or in a general articles in the newspaper. only two teachers surveyed had ever received any formal training with animal-assisted therapies, however most (80.8%) indicated they would be interested in attending a training session. motivators for integrating companion animals into the classroom when asked why they would consider involving animals or animal-assisted therapies as an intervention for students with asd, teachers gave a range of motivators. animals were identified as being able to offer something unique to the therapeutic and/or classroom environment. that is, animals resonated specifically with children with asd, and could be useful in a wide variety of contexts. “have always believed that animals have a way of reaching children that don't always respond well to adults or their peers” “animals do not discriminate nor judge others and can be trained to provide specific therapy to individuals. they bring out the best in people and give teachers more insight into students strengths” “animals generally provide an avenue to encourage asd children to actively participate in the world, rather than existing in retreat/ solitude” a personal interest in the intervention was expressed, as was a willingness to try new ‘things’ or ‘methods’ that might benefit their students. some took this further, indicating that it was their responsibility to find the most effective methods for individual students, but recognising that animal-assisted interventions may be effective for some, but not all. “i work with many children with asd. each child is different and as a teacher, it is my aim and responsibility to discover what works for each child to better their learning experience” “it depends on the student and their situation but if they like animals it might be a different perspective to helping them” there were some teachers who were already convinced that animal-assisted interventions are effective, and “been proven to work”. the rationale behind such strong views were not explored further. however, in a few cases, teachers had witnessed animal-assisted interventions being used effectively, and wanted to try it out with their own students. “i have visited a school where they have two golden retrievers as school pets and i was very impressed at the calm that they bought to a classroom and the obvious affection that the students had for them” perceived benefits of integrating companion animals into the classroom teachers who had introduced companion animals into their classrooms were prompted to share their experiences and provide brief examples of success. all reported positive experiences, and described a range of benefits to individual students as well as the classroom as a whole. perceived benefits of animal-assisted interventions fell into four distinct themes, including: page 17 attitudes of australian school teachers towards animals in the classroom creative common license 4.0 – non commercial – share alike – attribution smith, d.p. and dale, a.a. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science improving social skills. a variety of the social deficits that affect individuals with asd were reported to have improved considerably when animals were present. “asd students often show a strong bond with their pets and the ability to be open and expressive with these animals” “the dog taught an autistic child to play fetch. the child previously didn't interact or play with people and was afraid of animals” “teaching students turn taking skills and engaging students with animals and treating animals with respect” facilitating student engagement. it was noted that particular students were more likely to be focused and engaged in learning and participate in the classroom and curriculum based activities when animals were present. “kids love animalsyes, it’s a generalization but ‘generally’ they do. the children in my class would talk all day, if i let them, about their pets and their antics. they all bought along pet photos” “positive opportunity for teachers to engage children in games and fun and to enhance their participation and willingness to read” decreased student stress and anxiety. without a feeling of connectedness to the learning episode, peers or teacher, the classroom often becomes an environment where stress and anxiety, as well as problem behaviours rise and interfere with learning and social interaction. “students were drawn to the animal and displayed decreased anxiety and arousal. this calming nature allowed good discussion time and focus for students” “a student was very anxious and upset as he has not been picked up at school to go home. minty (animal assisted therapy dog) calmed the student while waiting” reducing problem behaviours. in addition to a decrease in stress and anxiety in students with asd, a decrease in problem behaviours were also noted. “i believe animals can transcend minor difficulties with mood and behaviour as they have keen senses and naturally helpful attitudes” “therapy dog worked with one of the ots [occupational therapist] in the school. was an incentive for students with behavioural concerns. dog made some children very happy” perceived barriers to the integration of animals into the classroom despite the encouraging examples provided by those who had experience with companion animals in the classroom, the teachers in this sample identified multiple barriers that they either have experienced, or expect to experience regarding the successful introduction of animals within their school and or classroom. some key logistical issues that prevented the use of animals included the employment status of the teacher (i.e., casual or part time teachers reported that it were difficult to instigate any animal related programs), and where animals were perceived not to be of obvious benefit to the subjects or classes being taught (e.g. maths, history and geography teachers felt animals would not provide any additional benefit). employment status and subject aside, several concerns or perceived barriers were raised that could be grouped according to four major themes: a lack of knowledge. a common barrier identified by teachers was a lack of knowledge about animal-assisted intervention, as well as how to access suitable information. of particular concern, was the lack of evidence base for which to support the decision to introduce animals (or animalrelated therapies) into the classroom. “if research showed that aat [animal assisted therapy] could have a positive benefit for my students with asd i would be willing to try it” page 18 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 13 22 “i think it is a developing area and evidence based practices are required before a higher level of implementation occurs” a lack of support and facilities. teachers felt that they would not have the support of their school if they were to enquire about implementing animal-assisted interventions. this includes resources (space, facilities, and funding) as well as support from administration. “i think it’s a great idea, especially if the program could focus on providing information to principals and getting them 'on board'” animal welfare concerns. teachers expressed concern for the well being of the animals when interacting with the children. “dog was overwhelmed by students who had not yet been able to have their own needs met and therefore were unresponsive to the animals needs” in addition, animal-assisted interventions were likely to cause additional work beyond the classroom, particularly the adequate care for the animal outside of school hours. “need to have a teacher that is willing to take an animal for training and house the animal. not very easy with most staff already having animals” “issues arose when the dog was sick and required careful management and support for students and staff and when therapy dog had to be retired” student behavioural reactions and allergies. another apprehension surrounded the reaction of the students to the animals. teachers felt that not all students would react well to having an animal in the classroom. “kids with asd can be just as unpredictable as a frightened dog. there are many factors that affect the success of a program like this” “…need to get parental approval and ensure there is not background of trauma with animals” of equal concern was the spread of disease and allergies of the children. “allergies are a major concern. i am very allergic to most animals and i would imagine many of our students could be.” “due to the nature of high schools as students change classrooms every hour, concerns for the almost certain spread of bacteria affects allergies” 4. discussion this study explored the attitudes and experiences of australian school teachers’ towards the use of animalassisted activities as a classroom based intervention for students with autism spectrum disorder. despite limited knowledge, training, or experience with more formal therapies, teachers showed a positive attitude toward, and a high interest in pursuing the inclusion of animal-assisted interventions. their interest was fuelled by a personal interest in animals; having previously witnessed the effective use of animals in the classroom; the belief that animals bring something unique to the classroom environment; and the belief that children with asd often resonate with animals. the teachers that had experience with animals in schools reported a range of benefits at the individual student and classroom level. the key benefits identified included improving social skills; facilitating student engagement by providing a means for the student to connect with the activity, teacher and peers; decreasing student stress and anxiety; and reducing classroom behavioural problems. these benefits mirror the positive teacher experiences reported elsewhere (hergovich et. al. 2002; kotrschal and ortbauer 2003; rud and beck 2003; prothmann et al. 2009), as well as studies identifying the benefits of children with asd interacting with animals (o’haire et al. 2013). of course, the use of animals or animal-assisted interventions in the classroom is not appropriate for all students (e.g. not all students like dogs, allergies), teachers (e.g. employment status), or situations (e.g. school, classroom setup, subject). explanations as to page 19 attitudes of australian school teachers towards animals in the classroom creative common license 4.0 – non commercial – share alike – attribution smith, d.p. and dale, a.a. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science why animal-assisted interventions were either not being adopted, or not currently considered included a lack of knowledge; lack of support and resources; extra workload; reactions of the student in relation to allergies and behaviour; and issues relating to animal welfare (both inside and outside of school hours). although these reflect some of the barriers reported by rud and beck (2003), and more recently ladarola et al. (2015), we found no indication that a lack of collegial or parental support was an issue for this sample of australian teachers. it is in the best interest of both schools and teachers to proactively source innovative interventions to support students with disabilities. overall, this study revealed that australian teachers are open to emerging or novel interventions to support students with asd. in fact, teachers expressed a personal responsibility to seeking the most effective teaching strategies for their students. as few teachers had actually used any form of animalassisted interventions in their classroom, it is important that the barriers identified to doing so are addressed. equally, resources that inform teachers, parents, and school administrators about animal-related programs must be made available. the lack of evidence-base from which to form a decision to implement animal-related interventions for students with asd was identified as an issue. the use of animals, although it may be incredibly beneficial for students, has yet to be supported by scientifically valid or reliable research in the classroom (o’haire 2013; kasari and smith 2013). thus, it is interesting that these teachers were willing to implement a time consuming and potentially financially costly intervention, despite a lack of evidence or training, and in a climate where it is often difficult to get schools to implement strategies and modifications that have been shown to genuinely make a difference (ladarola et al. 2015). indeed, it is important to keep an open mind in exploring different avenues, but there is an obligation to ensure that only interventions that have some empirical data and research base behind them are encouraged. moving forward, it is imperative that the inclusion of companion animals and more formal based animal intervention programs or activities in classrooms be adequately evaluated, because implementing or promoting any strategies without evidence is problematic. this scoping study represents a small sample of australian teachers that relies on self-report. additionally, it utilised a sample of convenience that likely attracted teachers with a pre-existing interest in animal-assisted interventions. a wider sample from a broad range of schools might reveal different attitudes, experiences, and barriers. there also may be some confusion among participants as to the nature of animal-related interventions (for example, around the differences between ‘formal’ animal-assisted therapies, and ‘informal’ animal-assisted activities). although a definition of animal-assisted therapy was provided to the participants at the beginning of the survey, their understanding of these terms may have influenced their responses unintentionally. despite these limitations, this study provides a useful starting point for further investigations into novel interventions that can support our increasing population of students with asd. equally important however, is to consider the attitudes and perspectives of the schools and school teachers because they will have the greatest influence on its adoption and its success. 5. references american psychiatric association. (2013). diagnostic and statistical manual of mental disorders: dsm-5. washington, d.c: american psychiatric association. anderson, k.l., and olson, m.r. 2006. the value of a dog in a classroom of children with severe emotional disorders. anthrozoös 19: 35-49. doi: 10.2752/089279306785593919. ascione, f.r., and weber, c.v. 1996. children’s attitudes about the humane treatment of animals and empathy: one-year follow-up of a school-based intervention. anthrozoös 9: 188–195. doi: 10.2752/089279396787001455 australian companion animal council. 2010. contribution of the pet care industry to the australian economy. 7th edn. http://www.acac.org.au/pdf/acac %20report%200810_sm.pdf. accessed on november 16, 2015. beetz, a. 2013. socio-emotional correlates of a school page 20 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 13 22 dog-teacher-team in the classroom. frontiers in psychology 4: 1-7. doi: 10.3389/fpsyg.2013.00886 berry, a., borgi, m., francia, n., alleva, e., and cirulli, f. 2013. use of assistance and therapy dogs for children with autism spectrum disorders: a critical review of the current evidence. the journal of alternative and complimentary medicine 19: 73-80. doi: 10.1089/acm.2011.0835 braun, v., and clarke, c. 2006. using thematic analysis in psychology. qualitative research in psychology 3: 77– 101. doi: 10.1191/1478088706qp063oa carlisle, g.k. 2015. the social skills and attachment to dogs of children with autism spectrum disorder. journal of autism and developmental disorders 45: 11371145. doi: 10.1007/s10803-014-2267-7. chandler, c. (2001). animal-assisted therapy in counseling and school settings (report no. edo-cg01-05). greensboro, nc: eric clearinghouse on counseling and student services. (eric document reproduction service no. ed459404). daly, b., and suggs, s. 2010. teachers' experiences with humane education and animals in the elementary classroom: implications for empathy development. journal of moral education 39: 101-112. doi: 10.1080/03057240903528733 fine, a. ed. 2010. handbook on animal-assisted therapy: theoretical foundations and guidelines for practice (3rd ed). san diego, ca: elsevier. friesen, l., and delisle. e. 2012. animal-assisted literacy. childhood education 88: 102-107. doi: 10.1080/00094056.2012.662124 grandgeorge, m., tordjman, s., lazartigues, a., lemonnier, e., deleau, m., hausberger, m., and young, l. 2012. does pet arrival trigger prosocial behaviors in individuals with autism? plos one 7: 1-8. doi: 10.1371/journal.pone.0041739 grandin, t., fine, a. h., & bowers, c. m. 2010. the use of therapy animals with individuals with autism. in handbook on animal-assisted therapy: theoretical foundations and guidelines for practice (3rd ed), 247–264, ed. a.h. fine san diego, ca: elsevier. green, j., willis, k., hughes, e., small, r., welch, n., gibbs, l., and daly, j. 2007. generating best evidence from qualitative research: the role of data analysis. australian and new zealand journal of public health 31: 545–550. doi: 10.1111/j.1753-6405.2007.00141.x hergovich, a., monshi, b., semmier, g and ziegler, v. 2002. the effects and presence of a dog in the classroom. anthrozoös 15: 37-50. doi: 10.2752/089279302786992775 jalongo, m., astorino, t., and bomboy, n. 2004. canine visitors: the influence of therapy dogs on young children's learning and well-being in classrooms and hospitals. early childhood education journal 32: 9-16. doi: 10.1023/b:ecej.0000039638.60714.5f jalongo, m. 2005. what are all these dogs doing at school? using therapy dogs to promote children's reading practice. childhood education 81: 152-158. doi: 10.1080/00094056.2005.10522259 kaufmann, m. 1997. creature comforts: animalassisted activities in education and therapy. national education service 1: 27-31. kasari, c., and smith, t. 2013. interventions in schools for children with autism spectrum disorder: methods and recommendations. autism 17: 254-267. doi: 10.1177/1362361312470496 kotrschal, k. and ortbauer, b. 2003. behavioural effects of the presence of a dog in a classroom. anthrozoös 16: 147-159. doi: 10.2752/089279303786992170 kršková, l., talarovičová, a., and olexová, l. 2010. guinea pigs—the "small great" therapist for autistic children, or: do guinea pigs have positive effects on autistic child social behavior? society and animals 18: 139-151. doi: 10.1163/156853010x491999 page 21 attitudes of australian school teachers towards animals in the classroom creative common license 4.0 – non commercial – share alike – attribution smith, d.p. and dale, a.a. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science ladarola s, hetherington s, clinton c, dean m, reisinger e, huynh l, locke j, conn k, heinert s, kataoka s, harwood r, smith t, mandell ds, kasari c. 2015. services for children with autism spectrum disorder in three, large urban school districts: perspectives of parents and educators. autism 19: 694703. doi: 10.1177/1362361314548078 lord, c., wagner, a., rogers, s., szatmari, p., aman, m., charman, t., dawson, g. et al. 2005. challenges in evaluating psychosocial interventions for autistic spectrum disorders. journal of autism and development disorders 35: 695-708. nimer, j., and lundahl, b. 2007. animal-assisted therapy: a meta-analysis. anthrozoös 20: 225-238. doi: 10.2752/089279307x224773 o'haire, m. 2013. animal-assisted intervention for autism spectrum disorder: a systematic literature review. journal of autism and developmental disorders 43: 1606-1622. doi: 10.1007/s10803-012-1707-5. o’haire, m.e., mckenzie, s.j., beck, a.m., and slaughter, v. 2013. social skills increase in children with autism in the presence of animals compared to toys. plos one 8: e57010. doi: 10.1371/journal.pone.0057010 o'haire, m.e., mckenzie, s.j., mccune, s., and slaughter, v. 2014. effects of classroom animal-assisted activities on social functioning in children with autism spectrum disorder. journal of alternative and complementary medicine 20: 162-168. doi: 10.1089/acm.2013.0165 prothmann, a., ettrich, c., and prothmann, s. 2009. preference for, and responsiveness to, people, dogs and objects in children with autism. anthrozoös 22: 161-171. doi: 10.2752/175303709x434185 roth, l. 2013. autism spectrum disorder, briefing paper no 5/2013. nsw parliamentary research service. isbn 978-0-7313-1901-5. retrieved from http://www.parliament.nsw.gov.au/prod/parlment/publ ications.nsf/0/b2142391f94516feca257b78001fb961/$f ile/autism%20spectrum%20disorder.briefing %20paper.pdf. accessed on november 16, 2015. rud jr., a.g., and beck, a.m. 2003. companion animals in indiana elementary schools. anthrozoös 16: 241-251. doi: 10.2752/089279303786992134 silva, k., correia, r., lima, m., magalhães, a., and de sousa, l. 2011. can dogs prime autistic children for therapy? evidence from a single case study. journal of alternative and complementary medicine 17: 655-659. doi: 10.1089/acm.2010.0436. smyth, c., and slevin, e. 2010. experiences of family life with an autism assistance dog. learning disability practice 13: 12-17. doi:10.7748/ldp2010.05.13.4.12.c7758. page 22 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 13 22 http://www.parliament.nsw.gov.au/prod/parlment/publications.nsf/0/b2142391f94516feca257b78001fb961/$file/autism%20spectrum%20disorder.briefing%20paper.pdf http://www.parliament.nsw.gov.au/prod/parlment/publications.nsf/0/b2142391f94516feca257b78001fb961/$file/autism%20spectrum%20disorder.briefing%20paper.pdf http://www.parliament.nsw.gov.au/prod/parlment/publications.nsf/0/b2142391f94516feca257b78001fb961/$file/autism%20spectrum%20disorder.briefing%20paper.pdf dogs trigger attention during animal assisted intervention in prison: a preliminary study 2021 vol. 11 1 14 dogs trigger attention during animal assisted intervention in prison: a preliminary study marine grandgeorge1, martine hausberger2, christine heyraud3, astrid hirschelmann4 abstract: animal‑assisted interventions (aai) seem to offer promising possibilities to prevent daily conditions of inmates (overcrowding or social isolation); however, nothing is known either about the potential processes involved or impact aai on the development of interactions between inmates. we hypothesized that either dogs would be a source and the centre of a ention, thereby that dog may induce more dog‑inmate interactions, or dogs would be social catalyst, i.e. facilitator of social interactions between humans. for that, we analysed first one‑hour aai sessions involving 10 adult male inmates, 7 service dogs and one dog handler. an observer recorded, using ethological methods, spatial distances between dogs and inmates and between humans, direction of inmates’ gazes and their vocal behaviour. hypothesis that dogs could be social catalyst was not supported: each inmate interacted mainly with his own dog. own dog was the almost only exclusive partner with whom they communicated: target of their visual gazes, vocal production and physical contact. based on literature and this preliminary research, we suggested that the animal/human ratio could be a crucial factor influencing the quality and quantity of aai interactions. m. grandgeorge*,1 m. hausberger2 c. heyraud3 a. hirschelmann4 1 univ rennes, normandie univ, cnrs, ethos (éthologie animale et humaine) ‑ umr 6552, f‑35380 paimpont, france 2 cnrs, univ rennes, normandie univ, ethos (éthologie animale et humaine) ‑ umr 6552, f‑35380 paimpont, france 3 univ rennes, normandie univ, cnrs, ethos (éthologie animale et humaine) ‑ umr 6552, f‑35000 rennes, france. 4 laboratoire de psychologie caen normandie, ea 7452, 14032 caen cedex, france *e‑mail: marine.grandgeorge@univ‑rennes1.fr highlights creative common license 4.0 – non commercial – share alike – attribution page 1 keywords: animal assisted intervention, prison inmate, visual a ention, interaction, group pet behaviour science 2021, vol. 11, 1 14 doi:10.21071/pbs.vi11.13330 creative common license 4.0 – non commercial – share alike – attribution www.petbehaviourscience.org • social isolation can occur even in overcrowded situations as social bonding can have been precluded • dogs here are not social catalysts, i.e. facilitator of social interactions between humans • dogs were the almost only exclusive partner with whom inmates had physical contact, preventing social isolation grandgeorge, hausberger, heyraud, & hirschelmannpet behaviour science creative common license 4.0 – non commercial – share alike – attribution marine grandgeorge martine hausberger christine heyraud astrid hirschelmann pet behaviour science 2021, vol. 11, 1 14 doi:10.21071/pbs.vi11.13330 introduction life in jail is characterised, amongst other aspects, by spatial and social restrictions (or overcrowding) that can lead to “a variety of health problems, injuries, and selected symptoms of psychological distress” (bonta & gendreau, 1990). it decreases self‑esteem (schni ker & john, 2007) and different factors have negative effects on inmates’ mental health (e.g. depression and increased risk of suicide) such as “overcrowding, various forms of violence, enforced solitude or conversely, lack of privacy, lack of meaningful activity, isolation from social networks, insecurity about future prospects” (e.g. work, relationships) (beynon & drew, 2001). the specific social organisation in jails, where both social isolation and overcrowding add to inmates’ supposed earlier social difficulties, is a major aspect of the difficulties inmates encounter in jail and it can impact their preparation for their future return to the « outside world » (beynon & drew, 2001). social isolation can occur even in overcrowded situations as social bonding can have been precluded. interestingly animal models provide insights concerning the processes involved: starlings, in the absence of social bonding despite being in a group, developed neurological disorders similar to those related to physical social separation, revealing that « psychological » social isolation can be as deleterious as « physical » isolation (cousillas et al., 2006). for example, young children lacking dedicated care and a ention develop atypical behaviours and cognitive and social deficits, e.g. children’s language development is slower when they are neglected (allen & oliver, 1982), inducing general delays in many domains (romanian orphans: kaler & freemann, 1994). spatial restrictions imposed on animals as well as social isolation or instability elicit « psychological disorders » such as stereotypic behaviours or depression‑like profiles, as for instance in horses (e.g. fureix et al., 2012; mills & mcdonnell, 2005) or primates (e.g. camus et al., 2013; lutz et al., 2003). in particular, individuals can become apathetic and indifferent to environmental including social stimuli (e.g. camus et al., 2013; fureix et al., 2012; fureix & meagher, 2015; rochais et al., 2016), but in many cases, they can work harder to obtain social stimuli, a need for any social species, especially humans (e.g. perret et al., 2015; søndergaard & ladewig, 2004). jail conditions and inmates’ past experiences could however prevent social stimuli to be perceived as positive. in order to improve well‑being and also cognitive and social skills, special procedures may be required. heterospecific interactions could help develop the intraspecific social skills of individuals with social difficulties (e.g. children with autism spectrum disorders, grandgeorge et al., 2012a). positive outcomes have been observed for socially deprived people (e.g. elderly living alone garrity et al., 1989), people for who human‑animal relationships can promote interactive behaviours increasing, for instance, visual awareness or seeking proximity/ contact (grandgeorge et al., 2017; grandgeorge et al., 2012b; hunt et al., 1992; mertens & turner, 1988). such observations have been the basis for the development of animal‑assisted interventions (aai) (grandgeorge & hausberger, 2011) which are based on triadic interactions between a professional, a human recipient and an animal. first dedicated to various human populations (e.g. with asd, alzheimer disease, or physical disabilities, page 2 nimer & lundahl, 2007), they have become popular in prisons as a potential tool for increasing wellbeing and stimulating social skills, the animal being here a potential social “substitute”. some studies have however focused on such interventions with inmates, highlighting that dog‑assisted interventions reduce recidivism and suggesting an increase of inmates’ patience, sense of responsibility, self‑esteem and self‑worth and decrease disciplinary records and tension that need to be further explore (for a review, see cooke & farrington, 2016; duindam et al, 2020). for example, thank to dog assisted interventions, inmates could “build an alternative anticriminal identity” (e.g. duindam et al, 2020, andrews et al., 2006, hill, 2018). however, nothing is known either about the potential processes involved or about the potential impacts these interventions on the development of intraspecific skills, e.g. interactions between inmates. only direct observations of inmates’ behaviours during aai sessions can help understand the potential role of this triadic situation. moreover, questions remain concerning how they should be performed: should each inmate be a ributed one particular dog so as to develop a specific bond with it or should the sessions be centered on one dog “shared” by a group of inmates? a recent study of aai with dogs and children with asd showed that the children’s visual a ention was triggered by the dog’s presence, especially when in a situation of “social rivalry” when the dog handler focused entirely on the dog (grandgeorge et al., 2017). another field of researches develop the concept of a unement and synchronization (i.e. coordination of behaviors between interacting partners; for review see duranton & gaunet, 2016). since first work of daniel stern and colleagues (1985) and development of studies about infant and mother’s affect a unement, numerous studies confirmed the importance of coordinate behaviour as crucial key for the infants development of social skills (e.g. tronick and cohn, 1989). to date, such concepts are extended to interspecific interactions between humans and dogs (duranton & gaunet, 2015). this synchronization is expressed, for example, throughout direction of gazes and behaviors linked to spatial distances (duranton et al., 2017). this human‑pet dog synchronization is frequently linked to increased affiliation and social responsiveness both in adults and children (e.g. duranton et al., 2017; wanser et al., 2021). interestingly, this behavioural synchrony exists during sessions of dog assisted interventions (pirronne et al., 2017). in the present study, we observed inmates’ behaviours during aai performed in a group with dogs, focusing especially on “social aspects” that could be linked to synchrony. aai programs in jail are based on the assumption that a dog can be a social catalyst, i.e. facilitator of social interactions between humans (messent, 1983) as observed in daily situations (hunt et al., 1992; mcnicholas & collis, 2000), especially if they are trained especially (eddy et al., 1988). however, given that in the present case each inmate was allocated one dog, we hypothesised that these animals would be a source and the centre of a ention, thereby potentially improving inmates’ well‑being (brickel, 1982). to evaluate the influence of aai on inmates’ well‑being, we developed an ethological approach involving direct observations, focusing on key aspects of social cognition: visual and vocal a ention as well as inter‑individual spatial distances (blois‑heulin & girona, 1999; feh, 2005; hausberger & cousillas, 1995; lemasson et al., 2003; mason, 1976; seyfarth, 1977). 2021 vol. 11 1 14 www.petbehaviourscience.org creative common license 4.0 – non commercial – share alike – attribution pet behaviour science 2021, vol. 11, 1 14 doi:10.21071/pbs.vi11.13330 page 3 marine grandgeorge martine hausberger christine heyraud astrid hirschelmann material and methods study location and dog‑assisted program. the study was performed in a french jail unit (condé‑sur‑sarthe, france) from february to may 2016. this correctional institution is especially for adult male offenders with very long sentences (over 20 years). this jail offers different types of special treatment programs to help inmates prepare to reintegrate society. the dog‑assisted program was one of these programs and was proposed on a voluntary basis to all motivated male inmates who do not fear dogs and a er an interview with their integration and probation officer. this dog‑assisted program was conducted in a specific room in the jail unit and was a group activity as 2‑4 inmates were present simultaneously during the sessions in addition to the dog‑handler (always the same woman). each inmate was allocated a service dog by the dog‑handler in accordance with the subjectively assessed inmate’s and dog’s temperament (i.e. 2‑4 service dogs were present simultaneously). inmates were asked their preference and in our study dog handler and inmate choices were similar. the dog‑handler interacted with the inmates with an open‑minded a itude and was unaware of the inmates’ life history unless an inmate chose to inform her. each session lasted one hour and there was one session per week. the intervention was semi‑structured, with at least three 20‑min activities during each session, namely grooming dog (e.g. brushing), feeding and finally walking it. between activities, inmates were free to occupy themselves otherwise (e.g. inmate talked to his dog or played with a ball). ethical concern regarding service dogs, the study was conducted in accordance with the french regulations governing the care and use of animals. these animals were not research dogs but were under the supervision of an accredited dog‑handler. the research was observational and did not influence dog treatment. all participants gave their free and informed consent to participate. all human‑related methods were performed in accordance with the declaration of helsinki (6th revision) and french regulations. all procedures were approved by the penal multidisciplinary commi ee, including the jail administration (commission pluridisciplinaire unique). all data were anonymous and neither video nor audio recordings were performed. participants inmates only inmates imprisoned for a long sentence (i.e. more than 10 years) and who had been in prison for at least 5 years were included a er having given their consent. eight male inmates (mean age of 38±5.0 years old, min‑max: 28‑51 years old), with a mean sentence of 24.7±3.8 years (range: 12‑30 years) participated in this study. although pet ownership or an interest for animals were not criteria for inclusion, all except one inmate had had at least one pet (i.e. pet behaviour science creative common license 4.0 – non commercial – share alike – attribution pet behaviour science 2021, vol. 11, 1 14 doi:10.21071/pbs.vi11.13330 page 4 grandgeorge, hausberger, heyraud, & hirschelmann marine grandgeorge martine hausberger christine heyraud astrid hirschelmann cat and/or dog) before going to jail and said they were interested in animals. to our knowledge, none was diagnosed with psychiatric or developmental disorders, but all presented relationship disorders. service dogs seven service dogs participated in this dog‑assisted program: 3 males and 4 females (mean age±sd: 18.9±2.9 months, from two breeds: golden and labrador retrievers). they were provided by the handi’chiens association where they had been trained appropriately and their behavioural evaluated (www.handichiens.org for more information). all service dogs had received the same training, which limited individual behavioural variations during sessions. dog‑handler a 40‑years‑old female dog‑handler working for the handi’chiens association animated all dog‑assisted sessions. she had 20 years’ experience of animal assisted interventions as a professional dog handler and had been working with inmates regularly for several years. experimental design the present study focused on the first aai session performed with each group of inmates, i.e. 2 groups of 4 inmates and one of 2 inmates. groups were composed according to inmates’ jail zone (i.e. all inmates in a group were familiar with each other). each session lasted one hour. dog aai material was available, i.e. leash, ball, brush, chair and floor mat. at the beginning of a session, each inmate was introduced to his service dog by the dog‑handler. all sessions were observed by the same female observer who was unfamiliar to inmates (only one encounter before to explain the research and to obtain their consent). data collection and analyses to respect correctional institution rules, no video recording material was used. all data were recorded through direct observation by the same observer, previously trained for coding interactions directly using ethological sampling methods (altmann, 1974: instantaneous scan sampling at 2‑minute intervals to record, for each inmate, the following behavioural items: ‑ direction of inmate’s gazes (independently of behaviour): gaze directed either towards his service dog, another service dog, another inmate, dog‑handler, observer or environment. ‑ target of inmate’s vocal behaviour (independently of content): speech or vocalisation addressed to his service dog, another service dog, another inmate, dog‑handler or observer. it could be a word, a sentence or an onomatopoeia. ‑ spatial distance between inmate subject and his partners, i.e. his service dog, another closest service dog, closest inmate and dog‑handler. our spatial distance unit was one inmate’s arm length: physical contact (i.e. touch the target), proximity (i.e. one to two arm lengths), medium distance (i.e. three to 2021 vol. 11 1 14 www.petbehaviourscience.org creative common license 4.0 – non commercial – share alike – attribution pet behaviour science 2021, vol. 11, 1 14 doi:10.21071/pbs.vi11.13330 page 5 marine grandgeorge martine hausberger christine heyraud astrid hirschelmann four arm lengths) and farthest away (i.e. more than 5 arm lengths) instantaneous scan sampling yielded two types of data: (1) frequency (in % of scans) of the different behavioural items recorded (i.e. vocal and eye direction) and (2) frequency of time spent at a given distance category from the different partners (i.e. proximity). statistical analyses as our data did not fit a normal distribution, we applied nonparametric statistical tests (siegel & castellan, 1988), i.e. kruskall wallis and mann– whitney u tests to compare independent samples, friedmann and wilcoxon signed rank tests to compare dependent samples and spearman rank‑order correlation coefficient to compare strength and direction of associations between behaviours and inmates’ characteristics (e.g. age). power analysis was done using kendallʹs coefficient of concordance. these analyses were run using statistica so ware© with the accepted p‑level set at 0.05. results visual a ention (figure 1a) globally, inmates gazed most of the time at living partners, i.e. humans or dogs (in only 5.64±3.86% scans were gazes directed towards the environment). around half of inmates’ visual a ention was focused on their own dog (45.84±5.38% scans, kendall w=0.779, friedman test=38.64 p<0.001). inmates gazed at the dog handler for approximately a fi h of the session duration (21.49±6.96%) and spent more time gazing at the observer than at the other inmates (12.41±4.18% versus 3.53±0.64% respectively; wilcoxon test z=2.93 p=0.003). inmates paired with a dog gazed less at the other dogs than at their own dog (9.73±1.4%; wilcoxon test z=2.8 p=0.005). inmates gazed rarely at each other (i.e. other participants were gazed significantly less 3.53±0.64% than all dogs, observer or dog handler, all wilcoxon tests, p<0.01). vocal behaviours (figure 1b) first, during approximately half the session the inmates did not talk (47.19±11.09%; kendall w=0.779, friedman test p<0.001). their own dog was the privileged target of an inmate’s vocal u erances (53.09±17.98%, kendall w=0.728, friedman test=36.72 p<0.001). inmates directed their vocal u erances significantly more o en to the dog handler than to the observer, the other inmates or the other dogs (27.88±16.87% versus 9.35±6.65%, 3.09±2.21%, 6.58±6.29%, respectively; all wilcoxon tests, p<0.01). pet behaviour science creative common license 4.0 – non commercial – share alike – attribution pet behaviour science 2021, vol. 11, 1 14 doi:10.21071/pbs.vi11.13330 page 6 grandgeorge, hausberger, heyraud, & hirschelmann marine grandgeorge martine hausberger christine heyraud astrid hirschelmann spatial distribution and physical contact subjects spent most of the duration of the session in direct contact or close proximity (i.e. <2 arms lengths) with their allocated dog that was almost exclusively the only partner with whom they had any physical contact (kruskal wallis test h=33.11, <0.001). subjects kept farther away from the other dogs, mainly at medium distance (friedman test=22.44, kendall w=0.762, p<0.001) and from their closest inmates (medium: 44.22±6.88%, and farthest: 35.77±8.23%, friedman test=23.48, kendall w=0.58, p<0.001). in addition, inmates kept far from the dog handler (farthest: 69.76±7.07%, friedman test=28.92, kendall w=0.749, p<0.001) figure 1. direction of (a) the inmates’ gazes (frequency of number of scans) and (b) the inmates’ vocal u erances, towards their own dog, another dog, another inmate, the dog‑handler, the observer or the environment (for visual a ention only). wilcoxon tests, p<0.05 when le ers differ. 2021 vol. 11 1 14 www.petbehaviourscience.org creative common license 4.0 – non commercial – share alike – attribution pet behaviour science 2021, vol. 11, 1 14 doi:10.21071/pbs.vi11.13330 page 7 marine grandgeorge martine hausberger christine heyraud astrid hirschelmann discussion their allocated dog appeared to be a privileged partner for the inmates, as their dog was their most frequent visual and vocal target, and was almost exclusively the only partner with whom they had any physical contact. thus, our results support our second hypothesis: inmates seemed to interact significantly more with their own dogs to the detriment of interactions with other dogs and or humans (supposed throughout few gazes and vocalizations). thus, the hypothesis that a dog could be a social catalyst (messent, 1983) especially dogs trained for being service dogs (eddy et al., 1988) seemed not supported by our data. that dogs can be privileged partners of interactions (i.e. vocal u erances, gazes and physical contact) is not specific to our study. indeed, dogs are powerful a ractive partners and can be a source and centre of a ention (brickel, 1982). during their first encounter where several species were present, young children are a racted to the dog, seek physical contact with it and address it vocally (nielsen & delude, 1989). when they have the choice, children with asd interact more frequently and for longer with an unknown dog than with an unknown human or toy (prothmann et al., 2009). nevertheless, the dog handler was a target of some inmates’ behaviours. one may argue that the social catalyst effect of a dog could then be involved (mcnicholas & collis, 2000). however, we must keep in mind the distinctive position of the dog handler, i.e. the person who knows the dogs and the inmates. being familiar with them could provide an optimal opening for inmates to exchange about the dogs and to interact be er with the dogs. the other humans were not the target of such behaviours, as it would have been the case if the dog played the role of social catalyst. if the dog played the role of social catalyst the other humans would have been the target of similar behaviours, but this was not the case here. our study focused on the first aai session, when all participants first met each other, the dogs and other humans in this context. this is first step in group building when affiliative dyads are created (hinde, 1979). here, at the beginning of aai one inmate was a ributed one dog that became his “own dog”. one could imagine that once, aai session a er aai session, a subject‑dog relationship is established, interactions and then relationships with other individuals would develop more easily, group cohesion would become stronger, even at a triadic level (simmel & wolff, 1964), as observed in other animal groups (mason, 1976; matsuda et al., 2012). if inmates and dogs synchronize their behaviours during aai, gaze to each other and be in physical contact, humans may benefit from these interactions (duranton & gaunet, 2016; nagasawa et al, 2009; pirronne et al., 2017). nevertheless, we could not be sure that the relationship network never got beyond a strong dyadic level between one inmate and his own dog as, for example, no specific competition appears between them (e.g. social rivalry, grandgeorge et al., 2017; schneider & krueger, 2012). the composition of a group is crucial for the development of social skills. even if the mother remains the first social model for many animal species to help learn social rules, other members of the group also appear important and so is pet behaviour science creative common license 4.0 – non commercial – share alike – attribution pet behaviour science 2021, vol. 11, 1 14 doi:10.21071/pbs.vi11.13330 page 8 grandgeorge, hausberger, heyraud, & hirschelmann marine grandgeorge martine hausberger christine heyraud astrid hirschelmann the adult/young ratio (bertin et al., 2007; bourjade et al., 2009). one could argue that the animal/human ratio would have a crucial influence on the quality and quantity of interactions. for example, the results of an aai with one dog and several elderly people indicated that direct physical contact with the animal remained limited but elicited other behaviours, i.e. gazing at the dog and at the other human participants as well as increased vocal exchanges between humans (olsen et al., 2016). as imprisonment is a highly stressful event (holmes & rahe, 1967) that can lead to “a variety of health problems, injuries, and selected symptoms of psychological distress” (bonta & gendreau, 1990), we could expect that time spent in prison or at least length of sentence would have an effect on inmates’ behaviours. however, this was not the case here, only inmates’ age had an effect on gazes and spatial distribution. older inmates focused more on their own dog and less on the environment (including observer), suggesting that – for them – their own dog was a more important source and a centre of a ention (brickel, 1982). this is interesting as, when adults encounter an unknown cat, seeking physical contact is not the only way to interact (mertens & turner, 1988), but strategy could depend of the animal species (nielsen & delude, 1989). to conclude, these direct observations are the first step towards understanding the potential processes associated with benefits of aai in jail (for reviews see cooke & farrington, 2016; duindam et al, 2020). to our knowledge, nothing is known about differences of benefits between aai performed in a group or individually, or about the influence of the animal/human ratio or species chosen. should each inmate be a ributed one particular dog so as to develop a specific bonding with it or should the sessions be centered on one dog “shared” by a group of inmates? here, it appears that the ratio one inmate/one dog elicited more interspecific that intraspecific a ention from the inmate point of view. although the question of whether aai strengthens the social bond remains unresolved, the mediated relationship with an animal may have a greater impact on psychological well‑being, which is part of the individualʹs primary needs. in this respect, this research opens up prospects for accompanying the period of execution of sentences that are in accordance with the good lives model theories that emphasize the involvement of all external and internal elements that strengthen human capital and make desistance a positive and consciously formulated life choice (ward & brown, 2004). in this context, the ʺprogramme de prévention de la recidiveʺ (criminal recidivism prevention program), published by the french prison administration, invites to build prevention programs which consist in bringing together a group of persons (convicted or pre‑trial) presenting a common problem, linked to the type of offence commi ed. the aim is to use group dynamics and the use of educational tools to make participants think about the consequences of their behaviour, get them to know themselves be er and give them the opportunity to adapt their behaviour to the rules of life in society. initiatives that enhance the quality of exchanges with others are all the more important because life in jail is characterised, amongst other aspects, by spatial and social restrictions (or overcrowding). at last, the aai may contribute to enabling the individual to invest the animal and through it a positive identity by opening up a field of 2021 vol. 11 1 14 www.petbehaviourscience.org creative common license 4.0 – non commercial – share alike – attribution pet behaviour science 2021, vol. 11, 1 14 doi:10.21071/pbs.vi11.13330 page 9 marine grandgeorge martine hausberger christine heyraud astrid hirschelmann possibilities towards the achievement of its fundamental needs. helping the individual to achieve his or her primary goods would therefore allow the individual to flourish and achieve a higher level of psychological well‑being, which is not a neutral effect given the prison context. we agree that our study has limitations (e.g. sample size, no longitudinal observations, no video recordings) that could not be overcome as they are inherent to prison conditions. for example, we were unable to study synchrony between inmates and dogs, as well as with others partners (both other dogs and humans) – or at least the cooperation and social interaction of the inmate with each other and with the dog. further studies need to be developed in the prison context as well as with other populations with disorders. nevertheless, extended contacts that can develop positive relationships are the best way to yield benefits from aai as well as from human‑pet bonds (melson, 2005). declarations funding not applicable conflicts of interest/competing interests the authors declare that they have no competing interests. availability of data and material not applicable code availability not applicable authorsʹ contributions a.h., c.h., m.g. designed the experiment, c.h. organized the population recruitment, c.h. collected the data, m.g., c.h. performed the analyses, m.g. and m.h. contributed to the statistical analysis, m.g., c.h., a.h., m.h. wrote the manuscript. acknowledgement we are grateful to the staff of both the handi’chiens association and the french jail unit at condé‑sur‑sarthe. we thank the participants, the foundation adrienne & pierre sommer for their support, veronique guyot for statistical help and the french gis ccs (groupement d’intérêt scientifque ‑ comportement cerveau et société). we are grteful to ann cloarec for the pet 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(2004). the good lives model and conceptual issues in offender rehabilitation. psychology, crime & law 10, 3, 243‑257. pet behaviour science creative common license 4.0 – non commercial – share alike – attribution pet behaviour science 2021, vol. 11, 1 14 doi:10.21071/pbs.vi11.13330 page 14 grandgeorge, hausberger, heyraud, & hirschelmann marine grandgeorge martine hausberger christine heyraud astrid hirschelmann h���������������������������� how old are (pet) dog breeds? christoph jung1*, daniela pörtl2 pet behaviour science | 2019, vol.7, 29 – 37 doi: 10.21071/pbs.v0i7.11494 christoph jung1*, daniela pörtl2 1.petwatch 2. psychiatric department, saaleunstrut klinikum, teaching hospital leipzig and jena universities, naumburg review * email: jung@petwatch.de germany keywords: dog; evolution; breed; coevolution; human-dogbonding highlights • dog breeds and even pet dog breeds have a long history • most of today's pet dog breeds were derived from working breeds • dog breeds may be understood as a reflection of human culture • we should understand dog breeds as continuously evolving populations in changing ecologies • understanding the history of breeds is crucial for a better understanding of dogs’ behavior and to improve human-dog-bonding page 29creative common license 4.0 – non commercial – share alike – attribution 2019 | vol. 7 | 29 37 abstract dogs are our pets. everybody knows dog breeds, at least we are familiar with common breads. a dog is often understood only as a specimen of a breed or a mongrel of several breeds. some scholars argue that dog breeds would be created as an artificial product starting 150 years ago in the victorian era. the original dog would be an uniform dog type called “village dog”, hanging around human settlements while scavenging human waste and faeces. our review of the literature suggests, that there is limited data and evidence relevant to the evolution and history of dog breeds and dog breeding in general to support this theory. in our article we will examine further data and present additional evidence. we found many records in history, archaeology and genetics pointing out that dog breeds have a long history likely starting in prehistoric times or at least in antiquity. dog breeds are not static. we should understand dog breeds as steadily evolving populations in changing ecologies like each other species. dogs’ ecological positions were defined primarily by humans. we are able to identify and clearly differentiate dogs according to their breeds; each breed is fitting to its special niches. we are using dogs’ different traits for thousands of years. dogs always have and continue to have their jobs related to hunting-, herding-, sledding-partners or as pets. consequently, dogs have been moulded to fit to each job and accordingly. in the long run, dogs evolved through their changing jobs and according to continually evolving human civilisation. breeds have not been simply created. breeds did not derive artificially during some decades in the victorian era. victorian dog breeding culture only switched the focus from the behaviour to the appearance and that mainly with regard to fashion dogs. even standardized modern purebred dogs on the official shows are continuously changing their traits and appearance following human fashions. dog breeds may be understood as a reflection of human culture. understanding the history of dog breeds is helpful for a better understanding of our dogs, the human-dog bonding and ourselves. http://www.petbehaviourscience.org/ introduction over 350 dog breeds are described all over the world. they are recognized by at least one of the big international kennel clubs. people love mongrels or their special breed. pet dog breeding has become a big business for over 100 years. however, we do not really know very much about the origin and history of dog breeding and dog breeds. some scholars spread the idea that dog breeds would be a relative new phenomenon in the evolution and history of dogs and humankind. they declare, that the era of dog breeding at first started in the victorian britain, 150 to 200 years ago (coppinger and coppinger 2003, 2016; lord et al. 2013; hekman 2018) as simply an invention of this era (worboys et al. 2018). they argue that village dogs would have been the original dogs, not living together with people but only nearby humans, scavenging on human waste. today's breeds would have been shaped due to artificial selection from village dogs imposed by fanciers during the last less than 200 years. our aim is to shed a light on the history of dog breeding and consequently human history and evolution as well. dogs are surely the first non-human animals living with us for at least 15.000 years (thalmann et al. 2013; botigué et al. 2017). we think it is important to study the evolution of dogs as a part of our own evolution and human culture. the question we would like to address and analyse: is it the nature of dogs hanging around human settlements as scavengers like most scholars and media argue? alternatively, was the relationship between dogs and humans a partnership? were dogs and humans living and “working” together already in ancient times (brewer et al. 2001; pörtl and jung 2017; jung and pörtl 2018; germonpré et al. 2018)? if so, dog breeds must have evolved a long time ago. we recognize that dog breeds evolved as a manifestation of optimized traits for their diverse jobs catering to humans, including hunting-, herding-, guarding-, sledding-partners or simply as pets. assigned tasks and jobs were, however, changing. dogs and people have shaped each other and impacted on each other permanently during their evolution and history. from the chihuahua to the great dane, dogs have been separated in breeds to fit to their different jobs. thus, they are quite different in appearance and behaviour. nevertheless, all dogs belong to exactly one species (canis familiaris) or sub-species (canis lupus familiaris). belonging to one species only the plant species cabbage shows so extensive different shapes like dogs do. can this be the fast result of diversification during only the last 150 to 200 years based on an uniform “original” dog type called village dog? can it be an artificial event? we find thousands of titles describing single pet dog breeds and their history in our book stores. quite all are written by breeders or fanciers of the described. recent breed monographs trend to idealize their special breed, often appearing like tales for marketing purposes. they are not suitable for an answer nor as a source of science. methods we adopted a broad multi-disciplinary approach including archaeology, paleo genetics, genetics and history. we investigated studies and ancient literature beginning with xenophon and aristotle and ending with a lot of dog books from the victorian era. interestingly charles darwin himself provides many hints and evidence for better understanding the nature and the history of dog breeds. results we found evidence towards the existence of a first prehistoric dog breeds specializing in polar bear hunting and also other breeds specializing in dog sledding. this research encompasses a time period of 9000 years (pitulko and kasparov 2017). ten fossilized dogs and a wooden sled were found on zhokhov island in the far north of siberia. the dogs surely could be separated in two types, seven appeared like a today’s siberian husky, and three like a today’s greenland dog. the authors assume, that “it can be hypothesized that sled dog teams might have been used in siberia as early as 15,000 years ago.” since the beginning of the neolithic period we found growing evidence for dogs as specialized working partners for hunting, herding, sledding, guarding in many regions page 30 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2018 | vol.7 | 29 37 (guagnin et al. 2018; perri 2016; jung 2011a). we know cave paintings and rock art from northern-africa or the arabian peninsula 9 to 10,000 years ago showing man and dog hunting or herding together (guagnin et al. 2018; coulson and campbell 2001; holl 2004). we have evidence for the development of specialized breeds at least starting during the first great civilizations like egypt, mesopotamia and greece (brewer et al. 2001). the ancient egyptians had been dog-lovers and bred at least ten documented breeds. it is remarkably that these breeds stood over all dynasties quite constantly, which means over 3,000 years (brewer et al. 2001; janssen and janssen 1999). about 2,350 years before present (bp), the great greek philosopher and scientist aristotle, named the founder of zoology, described seven dog breeds among them the maltese as a pet dog breed. aristotle gave hints how to feed and how to breed. "of dogs there are several breeds. of these the laconian hound of either sex is fit for breeding purposes when eight months old."(aristotle 2011, part 20). already 100 years before, another greek, xenophon, had published his book titled “cynegeticus” about hunting with dogs (xenophon 2017). we can understand this book as the first written collection of breed-standards with detailed dog breeding instructions. that means intentional dog breeding in ancestry. “cave canem” – it was common practice in ancient rome indicating that dogs guarded house and yard. that is not the typical job for a stray or village dog. the greek geographer strabo describes 2,200 years bp sophisticated dogs from the british isles: "britain produces hunting dogs with keen senses. the celts use these as their domestic dogs for war purposes." (strabo 2016) the roman geographer oppianus deepens this report and describes the british dogs as extremely strong and courageous fighters who could take on bulls (oppianus 2012). the germanic peoples compiled one article in their law, one from 22 articles, specially dedicated to dogs (lex baiuvariorum 2016). their law described eleven dog breeds. a good "leiti-hund" (like today’s bloodhound) had the legally based value of a horse. in 1868 charles darwin summarized the recent evolution of dog breeding: "as several breeds of the dog have been slightly but sensibly modified within so short a period as the last one or two centuries, by the selection of the best individuals, modified in many cases by crosses with other breeds; and as we shall hereafter see that the breeding of dogs was attended to in ancient times, as it still is by savages, we may conclude that we have in selection, even if only occasionally practiced, a potent means of modification." (darwin 1868, p. 43) intentional breeding is well documented from the middle age to the victorian era mainly concerning hunting and companion dogs (russell 2018). it was a privilege of aristocracy to breed hunting dogs. in the middle age, around 1387, the french count gaston fébus wrote a book about hunting in which the different types of hunting dogs, their use, their training, ailments and care are described in detail (klemettilä 2015). royals, aristocrats, and large landowners were competing for the best-specialized hunting dogs for each hunting tasks. thus, the great majority of dog breeds were and still are sophisticated breeds, specialized in one hunting profession, for example greyhounds for hounding game, beagles or foxhounds as pack hounds for fox hunts, pointers to find and point out the den of birds and hares, dachshunds for hunting the badger out of his hole. the aristocratic breeding farms (in germany called "jägerhöfe") were working very professionally with standards, pedigrees and stud books. lord orford, was the founder of the first ever coursing club, swaffham coursing society in 1776. he was respected as a very systematically working greyhound breeder, well documented not only by charles darwin (darwin 1860; russell 2018). not only hunting dogs have been bred intentionally for hundreds and thousands of years. interestingly, several pet dog breeds have a long and well documented breeding history. the maltese dog is recognised since aristotle. the pug or the blenheim spaniel were several times cited by comte de buffon (2009) and charles darwin (darwin 1860, 1868). konrad gessner (1551) and john caius (1570) described 21 british dog breeds in “de canibus britannicis“ (gessner 2008; caius 2018). comte de buffon (2009), the great french naturalist, mentioned 30 races in his “histoire naturelle” (1753 to 1767). georges cuvier, called the founder of modern zoology, mentioned in 1817 breeds like bulldog, page 31 jung & pörtl creative common license 4.0 – non commercial – share alike – attribution how old are (pet) dog breeds? https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science greyhound, pug, spaniel, barbet, terriers, pointer, great dane (cuvier 2012). in 1846 william youatt described 31 breeds, varieties and crosses (youatt 2015). he noted basic breeding instructions and wrote “it is probable that all dogs sprang from one common source, but climate, food, and cross-breeding caused variations of form, which suggested particular uses; and these being either designedly or accidentally perpetuated, the various breeds of dogs thus arose.” john henry walsh, called “stonehenge”, described 66 dog breeds (36 hunting, 15 pet) and six cross-breeds (walsh 2009). the austrian zoologists leopold fitzinger (2017) described 134 breeds and its varieties (among them 35 hound and 35 hunting breeds) and furthermore crosses of them in his report for the imperial austrian academia in 1867. h.d. richardson (2018) described 86 dog breeds very comprehensively along with illustrations; among them six pet dog breeds and furthermore nine mongrels among them the bull-terrier; the latter became an official breed in 1936 (akc). he also decribed basic dog breeding instructions (1857). both, charles darwin and richardson were documenting in detail and over seven generations the attempts to improve the braveness of greyhounds by crossbreeding them with bulldogs. in 1859, stonehenge mentioned that a greyhound pedigree should record 20 generations (walsh 2009). the dog breeds, charles darwin and others were describing explicitly and verbatim addressed as “breeds” or “races”, had never been "land races" or "village dogs". dog breeds were accurately described long before the kennel club was established in 1873. darwin and the naturalists of the mid 19th century strictly differentiated between mongrels and breed dogs and were discussing the effects of crossbreeding. this denotes the existence of breeds. consequently, we may hypothesize that intentional dog breeding has a very long tradition in human civilization, and did not suddenly start with modern dog breeding organizations. nevertheless, the victorian era set a caesura in the history of dog breeding. corresponding to basic changes in economy (industrial revolution), society (urbanization) and further changes pertinent to human lifestyle, for example, hunting practices, the tasks for dogs changed accordingly. still, hunting dogs have always had the broadest diversity pertinent to dog breeds; until to date the majority of dog breeds belongs to this group. about 60 page 32 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2018 | vol.7 | 29 37 figure 1. 140 years pug evolution under the same standard percent of the recognized breeds by the fédération cynologique internationale (fci) are hunting dogs. among them we are aware of 35 pointing dog breeds and 65 scent dog breeds. in the 19th century, huntsmen started hunting alone with modern guns. they needed a single all-purpose gun dog. mostly ancient pointers, retrievers, and spaniels have been modified to fit to the new requirements. later on some new dog breeds have been created by cross-breeding to produce a modern all-purpose police and working dog like the dobermann pinscher or the german shepard. the ancient companion dog breeds like maltese, pug, blenheim spaniel experienced a boom now being available not only for aristocratic ladies but also for urban citizens. thus, in the end of the 19th century, breeders began to establish "new" breeds stemming from varieties of the original breeds. this, to boost their business especially concerning bichons like the maltese (aka bolognaise, havanese, coton de tuléar, löwchen) or small terriers like the scottish terrier (aka west highland white, skye, cairn terrier). in victorian britain, many breeds had lost their “old jobs” for several reasons. some had gone extinct. the "turnspit", a dachshund-like but very strong dog, was designed and intentionally bred to run on a wheel, called the turnspit wheel, to turn meat over the fire, to power spinning wheels, pumps and bellows over hundreds of years. his nickname was "the underdog". he has become unemployed because of steam engines and electric motors. after hundreds of years he has gone extinct very fast. in north america, the same destiny befell other dog breeds as well. the indigenous coast salish peoples (bc, canada) had a special woolly dog. his hair became yarn for particularly valuable blankets. labor-intensive dog-hair weaving disappeared quickly after the introduction of machine made blankets by trading companies. consequently, the woolly-dogs were reportedly extinct by the third quarter of the 19th century (barsh et al. 2002). the prohibition of dogfights with the “cruelty to animals act 1835“ was fortunately the destruction of the last bulldog jobs in the united kingdom. decades earlier, dog fighting had gone out of fashion. bulldogs had a very high reputation because of their courage and braveness, and was addressed as the british national dog. for that reason, urban gentlemen were looking for a bulldog as companion dog. clever entrepreneurs quickly recognized their opportunity. at least since 1825 breeders like ben white and bill george operated puppy mills to satisfy the big demand for the new pet bulldog (farman 2010). the bulldog was the first breed that had been converted from a working dog to a pet dog. later on, in 1873, it should become the very first breed of the victorian era officially recognized by the recently established kennel club. we all know what deep modifications this exactly “standardized” breed had to suffer through, taking not only our modern breeding system. we all know what a modest future the new system has provided for the bulldog (jung 2011b). the bulldog has been the very first, but not the only dog breed switching from a working to a companion or pet dog. starting with the bulldog in the 1820th over more than 150 years until today about 60 dog breeds basically changed their role that way. the victorian era marks the dawn of companion and pet dog breeding as a business. however, that are less than one-third of totally recognized breeds by the akc, the kc or the fci. until today, business people keep on trying to place working dog breeds as pet dogs to conquer a new profitable market. even breeds like livestock guarding dogs, e.g. kangals or ovcharkas or high-end working dogs like border collies have been offered as pet dogs. thus, many breeds have been splitted in a working and a show line to cover both markets. however, the majority of modern dog breeds still represent ancient working breeds, mostly used for hunting purposes. the victorian era was an era of standardization that began with the standardization of thread sizes to boost production and trade. official stud books for horses were starting in the dawn of the 19th century. british horses had been exported worldwide. some decades later british breed dogs followed up. thus, dog breeds have been standardized and uniformed as well. the spread of railroads enabled dogs from distant regions to mate, homogenizing their appearance on a national scale (russell 2018). this new breeding industry needed regulation and quality standards to improve business worldwide. fanciers demanded breed standards for the preservation of the old working breeds, which had lost their original jobs. the kennel club (kc) was founded on 4 april 1873. gradually, new guidelines were set. they are officially valid until page 33 jung & pörtl creative common license 4.0 – non commercial – share alike – attribution how old are (pet) dog breeds? https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science today: • registration for each dog for unique identification • stud books, closed or with appendix registries; stud books provide a record of results for all championship dog shows • pedigree for each dog (certificate for "pure bred") • written standards to establish a detailed image of the ideal appearance of each breed. "breed standards still reflect the reasons why the breed came into being" (the kennel club 2018) • championship dog shows in which (mainly companion not hunting/working-) dogs are rated for how well their appearance conforms to a standard • rating on the shows by independent judges • breeding clubs for each breed joined to the kennel club that is the theory. standardization in the 19th century did not lead to unification in dog breeding and each breed has steadily changed its appearance more or less depending on fashions. even the classification as breed sometimes seems to be arbitrary. as an example, 192 dog breeds are officially recognized by the american kennel club, 220 recognized by the kc and not less than 345 recognized by the fci on a so-called "definitive basis" (2018). from 192 to 345 officially recognized breeds, that is quite a difference and we mentioned only the three biggest international kennel clubs. that gives us another hint that the victorian breeding culture does not fit to the functional, zoological and historical view on dog breeds nor to evolution nor to ecology (figure 1). in particular, the principle of purebred breeding does not fit into reality and has never done so. in theory, we have closed stud books. even officially, we see open stud books or stud books with appendix registries. unofficially it is normal to fresh up the blood of a kennel from time to time by inter-breeding with phenotypically similar specimen or those from other kennel clubs. in particular, after the second world war, most companion and pet dog breeds had lost their breeding potential. there was a genetic bottleneck. thus, most stud books have been opened for many years. the real purebred dog is an idealized image and in some cases simply a breeders marketing tale. for dogs’ fitness that are good news. however, modern puppy mills are breeding their own way to gain maximal profit. the rules of kennel clubs do not matter. in germany, two out of three breeding puppies are produced in such farms without any control. especially young fanciers, buying their pups online, are not interested in basic breeding standards (packer et al. 2017). nevertheless, we can clearly identify a dog breed by appearance, behaviour and its genetic fingerprint (vonholdt et al. 2010; parker et al. 2017). given such a variety in appearance and behaviour, it is unlikely that several and well described dog types would have derived from uniform village dogs in just a few decades and until the official initiation of dog breeding in 1873 with dozens of modern breed standards to follow. dogs seem to live with human as specialized breeds since ancient times. we have found evidence that dog breeds and intentional dog breeding have been a very deep-rooted part of human culture – this evidence dates back for about 200 years. in his pioneering work (1860, 1868, 1874) charles darwin does not leave room for any doubt that dogs are separated in breeds and that these breeds have an ancient even “wild” origin. he described many dog breeds with an in each and classified them according to their different and clearly distinct behaviour including what he called “domestic instincts”, e.g. pointers, retrievers, shepherds (1860, p. 213). darwin assumed that „of another breed for another purpose; when we compare dog breeds, each breed seems to serve a purpose for humans in very different ways.” (1860, p. 34) and “no instance is on record of such dogs as bloodhounds, spaniels, true greyhounds having been kept by savages: they are the product of long-continued civilization.” darwin dedicates a separate chapter related to the question of “differences between the several breeds of the dog.” (1868, p. 33) page 34 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2018 | vol.7 | 29 37 the victorian era has set a caesura for numerous dog breeds. the focus was transferred from appearance towards of behaviour and fitness. the novel focus focused on “dog fashion” instead of dogs fitting to provide certain jobs. consequently, dog entrepreneurs introduced several policies pertinent to pedigree dog breeding, including dog shows. the victorian era primarily provided a framework for pet dog breeding as an upcoming international business not more. quite all dog breeds have reliable roots in ancient times. many – but not the majority – of dog breeds have been switched over a time window from 1825 (bulldog) until today (e.g. australian shepherd) from working to pet dogs. regulations taking place in victorian britain were the beginning of nationwide dog breeding policies, in particular for dog shows (russell 2018). nevertheless, it was not the dawn of the supported breeds. fashion dog breeding guidelines just adopted ancient dog breeding practices to the new requirements in modern human society and economy, their new ecological niche. only a few new breeds have been created until now. it is likely, that most breeds have been modified and every so often gave rise to several “new” breeds. that is the nature of breeds – not only concerning dogs. breeds are made for and by humankind. all breeds horses, sheep, grains, cabbages or dogs are designed for special purposes in special environments, climates and ecologies. purposes and ecologies are steadily evolving; thus breeds steadily adapted to new requirements. while some dog breeds adapted, some breeds became extinct (turnspit, woolly dog), some were modified (bulldog), some were created new (german shepherd), some were reinvented (hovaward) and some were recreated (irish wolfhound). given the above, we have evidence for highly heritable and functionally relevant breed differences in dog behaviour (mclean et al. 2019). for example, the victorian era of dog breeding simply introduced new policies to adapt classical intentional dog breeding to modern business purposes. nevertheless, even and in particular, under the policies of the victorian breeding culture like official standards, show-judges and stud books, dog breeds have been modified steadily. if we look only at what has been bred entirely under the rules of the british kennel club, we have to realize that many pet dog breeds have changed their appearance and their behaviour dramatically e.g. pug, bulldog, basset hound, german shepard and are still changing until now (fig. 1). conclusion and discussion we present evidence related to dog breeds, their origin and evolution long before the victorian era began. we also present data pertinent to intentional dog breeding dating back for at least 2,400 years (brewer et al. 2001). dog breeds evolved and were later purposefully bred to perform and optimally various tasks as ordered by man. hunting, guarding, sledding and herding are thought to be the earliest dog jobs. even to serve simply as a pet is an ancient purpose. the separation in breeds or races is a need and a fundamental mark for the dog as a human working companion and partner (jung and pörtl 2018). humans desired and thus bred a specialist for each job even for the job as pet. cultures and technologies developed, as a consequence jobs carried out by working dogs did so as well. we may understand dog breeds as a reflection of human cultural evolution and history. possibly, we may address it as a form of coevolution (russel 2018; schleidt and shalter 2003, 2018). darwin referred to this process as “coadaptation” (1860, p. 31). however, dog breeds are not and have never been set in stone (russel 2018). in particular the so called pure bred dogs under the kennel clubs like kc, akc and fci are changing their appearance steadily and sometimes fundamentally especially in the last decades. comparing the official show champions and the “best of breed” winners from 1970, 1990 and 2010 the differences are astonishing. if the traits of a breed change, it does not mean that it is a new breed now. humankind has formed every breed for its own purposes. this dates back to prehistoric times – including plants or animals. formerly, we have had dozens of clearly different tomato breeds for taste, climate, usage. therefore, we did not need the european union bureaucracy with its standardization and tomato specifications filling pages. nevertheless, today only very few and selected new eu tomato breeds are officially recognized in europe. the same is valid for dogs. not kennel clubs invented breeds but breeds “invented and shaped” clubs. dogs as breeds have not been invented in victorian britain. breeds did not derive during decades from village dogs. victorian page 35 jung & pörtl creative common license 4.0 – non commercial – share alike – attribution how old are (pet) dog breeds? https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science breeding simply standardized dog breeds by appearance. even dog breed standardization was much older and was not invented at that time (xenophon 2017; klemettilä 2015). only the criteria, the scope and the methods were and have changed. in former times and regarding working dogs until today primarily traits, behaviour and fitness were the leading features of each breed. in ancient times, breed standards had not been set up by references aristotle 2011. historia animalium. cambridge: cambridge classical texts and commentaries. barsh r. l., jones j. m. & suttles w. 2002. history, ethnography, and archaeology of the coast salish woolly-dog. in l. m. snyder and e. a. moore (eds.), dogs and people in social, working, economic or symbolic interaction (pp. 1-11). oxford: oxford books. botigué l. r., song, s., scheu a., gopalan s., pendleton, a., oetjens, m., taravella, a., seregély, t., zeeb-lanz, a., arbogast, r., bobo, d., daly k., unterländer m., burger, j., kidd, j., veeramah, k. 2017. ancient european dog genomes reveal continuity since the early neolithic. nat. commun. 8 16082. brewer d. et al., 2001. dogs in antiquity. anubis to cerberus. the origins of the domestic dog. warminster: aris & phillips. caius, j. 2018. de canibus britannicis. reprint outlook verlag. comte de buffon. 2009. histoire naturelle. reprint komet. coppinger, r. p., coppinger, l. 2001. dogs: a new understanding of canine origin, behavior and evolution. chicago: university of chicago press. coppinger, r. p., coppinger, l. 2016. what is a dog? chicago: university of chicago press. coulson, d., campbell, a. 2001. african rock art: paintings and engravings on stone new york: harry n abrams. cuvier, g. 2012. animal kingdom. reprint forgotten books. darwin, c. r. 1860. on the origin of species by means of natural selection, or the preservation of favoured races in the struggle for life. london: john murray 2nd edition. darwin, c. r. 1868. the variation of animals and plants under domestication. london: john murray. darwin, c. r. 1874. the descent of man, and selection in relation to sex. london: john murray 2nd edition. farman, e. 2010. the bulldog a monograph. vintage reprint dog books. fitzinger, l. j. 2017. der hund und seine rassen: naturgeschichte des zahmen hundes, seiner formen, rassen und kreuzungen. norderstedt: reprint verlag der wissenschaften. germonpré, m., láznicková-galetová, m., sablin, m.v., bocherens, h. 2018. self-domestication or human control? the upper palaeolithic domestication of the dog. in vigne, j.d., stépanoff, c. 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– attribution how old are (pet) dog breeds? https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science pet attitudes predicting preferences for pets over people 2022 vol. 13 16 31 pet attitudes predicting preferences for pets over people katherine aumer1, *, michael erickson2, jack krizizke3, marc jaksuwijitkorn4, jennifer åbb5 abstract: the preference for animal companionship over human companionship may be predicted by a itudes about pet ownership. we hypothesized that pet a itudes could predict preferences for relationships with pets over humans. we sampled 182 people who named a person and a pet they love and care about. participants rated their feelings of love, time spent, enjoyment, and equity in both their human and pet relationships. we also presented seven hypothetical negative event scenarios that involve both the pet and human and asked participants to predict their feelings and reactions based on these events. the pet a itudes scale (templer et al., 1981) was used to assess a itudes towards pets. people had similar positive feelings about their pet‑human and human‑human relationships. however, people were more likely to react negatively towards a human compared to a pet. positive pet a itudes predicted more positive and less negative reactions to pets. positive pet a itudes can predict preference for pet relationships over human relationships and may help researchers identify what relationships work best depending on a personʹs pet a itudes. highlights creative common license 4.0 – non commercial – share alike – attribution keywords: animal‑companionship; pet a itudes; relationships; pets pet behaviour science 2022, vol. 13, 16 31 doi:10.21071/pbs.vi13.13473 creative common license 4.0 – non commercial – share alike – attribution www.petbehaviourscience.org page 16 katherine aumer1, michael erickson2, jack krizizke3, marc jaksuwijitkorn4, jennifer åbb5 1 university of hawai‘i–west oʹahu. 91‑ 1001 farrington hwy, kapolei, hi 96707 2 university of california, riverside. 900 university ave, riverside, ca 92521 3 antioch university 2400 3rd ave #200, sea le, wa 98121 4 roosevelt university. 430 s michigan ave, chicago, il 60605 5 hawaiʹi pacific university. 1 aloha tower dr, honolulu, hi 96813 address for correspondence: address for correspondence: katherine aumer, 91‑1001 farrington hwy, kapolei, hi 96707. e‑mail: kaumer@hawaii.edu • as pet ownership increases and some report preferences of animal companionship over human companionship, it is important to identify key factors that are related to positive pet ownership. • a itudes towards pets and pet ownership was related to a preference of pets over human companionship, with more love and less negative responses aimed at pets versus people. • assessing a itudes towards pets and pet ownership may provide insight into animal vs. human companionship preferences and suitability for effectiveness of animal‑assisted therapy and companionship. aumer, erickson, krizizke, jaksuwijitkorn, & åbb pet behaviour science creative common license 4.0 – non commercial – share alike – attribution katherine aumer, michael erickson, jack krizizke, marc jaksuwijitkorn, jennifer åbb introduction both humans and pets can provide people with love and support. unlike non‑ domesticated animals, pets or animal companions have a special relationship with people that is a result of various historical and cultural norms (herzog & foster, 2010; serpell & paul, 2011). pets have helped with hunting, husbandry, prevention of pests or vermin, providing personal guidance, and are recognized in their ability to help humans medically and emotionally (see blouin, 2012 for a review). as discussed by veevers (1985) pets can also have a variety of emotional and social functions in the human‑pet relationship. for example, a pet may have a projective function, in which they help a human through their selection, breed, size, or other characteristics make a statement about the human and their social standing (hirschman, 1994; veevers, 1985). pets can also help serve as a kind of social lubricant, helping people get to know each other or bond because of their pet relationships (robins, sanders, & cahill 1991; veevers, 1985). pets can also serve as a surrogate, either supplementing or in some cases, providing an alternative to human‑human relationships (basten, 2009; belk, 1996; veevers, 1985; walsh, 2009). pets can serve many of these functions and more. of course, like any relationship, the human‑pet relationship does have costs: sharing of food (vale & vale, 2009), resources (pew research center, 2006), shelter (appma, 2008), time (voith, 2009), and sharing or spreading of disease (center for disease control and prevention, 2021). however, most of these costs do not seem to deter people from owning pets as over 57% of american families own at least one pet (strochak, goodman, & zhu 2018). some researchers have argued that pet or animal companion relationships share similar qualities as that between a parent and a child (franklin 1999; serpell 1996). pets may help humans provide an outlet for that desire to give and receive a achment love, while avoiding the costs and social challenges of raising a child (bha arai, 2017). some evidence suggests that people may find that growing their family with a pet may be more appealing than growing their family with a human child (cohen, 2002). for example, recent trends in pet ownership have shown that households with at least one pet are increasing while households with at least one child are decreasing (strochak, goodman, & zhu 2018). during the lockdown and stay‑at‑home orders of the covid‑19 pandemic, many individuals sought dogs and cats to add to their families (hedgpeth, 2021). however, adoption and fostering of children during the pandemic decreased (white & blackburn, 2020). some evidence has suggested that people actually prefer pet‑human relationships over human‑human relationships. for example, results of a survey by onepoll in 2019 suggested that not only do many see pets as people, but a majority of parents preferred their pets over their children (haaland, 2019). however, we have yet to find any scholarly research to support a preference for pets over human relationships. many studies, using interviews and qualitative analyses, were able to establish the strong bond between people and their pets (e.g., basten, 2009; laurent‑ simpson, 2017). additionally, observational studies as done by robins and colleagues (1991) and sanders (1990) have solidified the important role pets can play in people’s social lives with other humans. however, few studies have a empted to quantify these relational impacts. for example, dotson and hya (2008) surveyed over 700 dog owners about the underlying dimensions of dog page 17 pet behaviour science 2022, vol. 13, 16 31 doi:10.21071/pbs.vi13.13473 creative common license 4.0 – non commercial – share alike – attribution page 18 ownership and their measures established relationships between a person’s demographic characteristics and their likelihood to belong to certain dimensions related to the construct of dog‑ownership. however, dotson and hya ’s (2008) study does not help explain a possible preference for pets over people. several scales regarding pet commitment have been developed, including: the lexington a achment to pets scale (laps: johnson, garrity, & stallones, 1992) and the miller‑rada commitment to pets scale (cps: staats et al., 1996). these studies and scales provide evidence of the importance of pet‑ownership and animal companionship. the laps (johnson, garrity, & stallones 1992) is especially useful when measuring feelings, a achment, and anthropomorphism of pets. the cps (staats et al., 1996) has been used to measure commitment and care towards pets and predict behaviors in regards to treatment of pets (brackenridge et al., 2012). both of these scales provide quantitative measurements regarding the quality of pet relationships, however to be er understand the factors that contribute to preferences for pets over people we turn to a itudes. a itudes a itudes are global evaluations toward an object or issue (eagly & chaiken, 1998). whether one likes or dislikes pets is an a itude about pets. for the purposes of this study, if one has a positive a itude about pets then one generally likes pets, the relationship one has (had or could have) with pets, and taking care of pets. a negative a itude about pets would denote disliking pets, the relationship one has (had or could have) with pets, and taking care of pets. of course, one may have dual a itudes (both positive and negative) about pets and given our definition, pet a itudes could be measured as a distribution. a itudes are separate from a achment as one could theoretically have a strong a achment to an animal companion, but have a negative a itude about pets. cute pets and ones that express child‑like or human‑like responses or behaviors may be especially likely to evoke positive human reactions and positive a itudes (sherman & haidt, 2011). beyond cuteness, animals who have certain anthropomorphic traits may also be more likely to a ract the a ention and care of people in need of non‑human companionship and support (serpell, 2003). one may also argue that if a pet has a shared history with a person, then one could develop a more positive a itude about animal companionship and pets (sable, 1995). beyond positive feelings, there are practical and several adaptive benefits from animal companionship that may influence pet a itudes. animal companionship can increase feelings of social support which can impact the likelihood to survive and the fitness of the human owner (eriksen, 1994; serpell, 1991; serpell, 2003; serpell & paul, 2011). however, from an evolutionary psychology perspective, any preference for pet‑human relationships over human‑ human relationships is a bit puzzling, especially if one considers the preference of a pet over a human that one is related to genetically (archer, 1997). given the theory of kin selection (smith, 1964), one would suppose that people are much more willing to engage with and help others that one is related to or share genes with than those one does not (e.g., help feed and change the diapers of oneʹs own child or parent, but not necessarily the children or parents of others). however, pets or animal companions share considerably less genetic material with their owners than a child, parent, or significant other. so why might people prefer pets 2022 vol. 13 16 31 www.petbehaviourscience.org katherine aumer, michael erickson, jack krizizke, marc jaksuwijitkorn, jennifer åbb pet behaviour science 2022, vol. 13, 16 31 doi:10.21071/pbs.vi13.13473 creative common license 4.0 – non commercial – share alike – attribution page 19 over humans? a itudes may help explain this possible preference. predicting behavior from a itudes has been a long topic of debate in social psychology (for review see ajzen et al., 2018 and glasman & albarracín, 2006). however, there is some evidence that interest in or involvement with an a itude object, can be powerful in predicting behaviors (e.g., fazio & zanna, 1981). a itudes towards pets and pet ownership may provide some insight into whether one will rate their relationship with a pet as be er than that with a human. pet a itudes have been found to be related to a variety of relational outcomes. for example, planchon, templer, stokes, and keller (2002) found that those whose pet died were more likely to grieve if they had higher pet a itudes as measured with the pas, pet a itude scale (templer et al., 1981). additionally, those with more positive pet a itudes seem to receive even more benefit from medical assistance from animals, as those with more positive pet a itudes had lower blood pressure a er pe ing dogs (jenkins, 1986) and lower arterial pressure and systolic pressure a er pe ing horses (hama, yogo, & matsuyama, 1996). practical benefits and relational bonds aside, a itudes about pets in general may provide some indication for a preference for pets over human relationships. hypotheses this study measures the quality of relationships one has with both a loved and cared for pet and human and is intended to discover if pet a itudes are related to relationships with pets. we hypothesize that pet a itudes will be related to differences in pet‑human and human‑human relationships. specifically, people with higher (more positive) pet a itudes will be more likely to report more positive feelings (e.g., love, time spent, and enjoyment) with pets than with humans. similarly, people with strong pet a itudes will be more likely to report less negative reactions and feelings (e.g., hate, need to a ack, upsetness, and anger) towards pets than humans. we also explore differences in human and pet relationships, specifically: love, hate, time, and enjoyment. h1: people will report more equity with human relationships than pet relationships, because pets tend to take more time and resources than human relationships as previous literature has found. h2: higher scores on the pet a itudes scale will be positively correlated to higher reports of love for pets vs. human companions. h3: higher scores on the pet a itudes scale will relate to lower reports of hate for pets vs. human companions. h4: higher scores on the pet a itudes scale will relate to lower reports of upsetness, anger, and hate for negative scenarios for pets vs. human companions. aumer, erickson, krizizke, jaksuwijitkorn, & åbb pet behaviour science katherine aumer, michael erickson, jack krizizke, marc jaksuwijitkorn, jennifer åbb pet behaviour science 2022, vol. 13, 16 31 doi:10.21071/pbs.vi13.13473 creative common license 4.0 – non commercial – share alike – attribution page 20 method participants participants were recruited through social media and through the hawaiʹi pacific university subject pool. all methods have been reviewed and approved by hawaiʹi pacific universityʹs institutional review board (irb) protocol #560420042. participants were provided a consent form and if they consented to continue with the study were asked a series of demographic questions including age, race, gender, and location. we surveyed 182 people or which 103 (86 female & 17 male) were born and currently reside in the u.s. measures a er the consent form and demographics, participants identified both a human and a pet they currently love and care about (we allowed for relationships between humans or pets that have ended as it was assumed that people may still have feelings about this person or pet. the important part was identifying instantly, a person or pet that the participant loves and cares about.). the order of identification (human or pet) was randomized and balanced along with the following measures: relationship measures, negative event scenarios, and pas. participants identified the relationship with the human (e.g., parent, sibling, significant other, or friend) and the species of pet (e.g., dog or cat). for repeated measures anova analyses, sphericity assumptions were met. when normality was not met for one‑way anova, the welchʹs statistic is reported. relationship measures participants answered the same four relationship questions about their human and pet relationships including: (1) how much do you love (pet name or human name)? (2) how much do you hate (pet name or human name)? (3) how much time do you spend with (pet name or human name)? (4) how much do you enjoy spending time with (pet name or human name)? all of these questions were answered on a 1‑5 likert scale with 1 being “not at all/ none at all” to 5 being “completely/a lot”. participants also rated the equity in their relationship using the global measure of equity scale (traupmann, peterson, utne, & hatfield, 1981; young & hatfield, 2009) which asked one question: “consider what you put into your relationship, compared to what you get out of it, how does your relationship with pet/person stack up?” was answered on a +3 to ‑3 scale. positive values indicated ge ing a “be er deal” or being over‑benefited, a rating of 0 indicated “both ge ing the same deal” or equitable, and negative values indicated the “pet/person ge ing a be er deal” or under‑benefi ed in the relationship. to examine if pet a itudes were related to preferences for pet relationships, we used residual change scores for relationship and negative scenarios with human 2022 vol. 13 16 31 www.petbehaviourscience.org katherine aumer, michael erickson, jack krizizke, marc jaksuwijitkorn, jennifer åbb pet behaviour science 2022, vol. 13, 16 31 doi:10.21071/pbs.vi13.13473 ratings as the predictor and pet ratings as the dependent variable (jennings & cribbie, 2016). a negative standardized residual change score would mean that the rating was higher for the human and a positive standardized residual change score would mean that the rating was higher for the pet. we then regressed residual change scores across the relationship and negative scenario questions onto pet a itudes to see if pet a itudes could predict differences in relationship quality between human‑pet and human‑human relationships. negative event scenarios participants were presented with seven negative events scenarios and were asked to rate the degree of (1) upsetness, (2) anger, and (3) hate they would feel towards the human or pet if that scenario occurred. scenarios included: (1) if your (human/pet) a acked you, how likely would you a ack back? (2) if your (human/pet) destroyed something you loved, how much would you feel (upset/angry/hate)? (3) if your (human/pet) ignored you, how much would you feel (upset/angry/ hate)? (4) if your (human/pet) betrayed you, how much would you feel (upset/angry/ hate)? (5) if your (human/pet) treated someone else be er than you, how much would you feel (upset/angry/hate)? (6) if your (human/pet) ignored a gi you gave them, how much would you feel (upset/angry/hate)? (7) if your (human/pet) threatened you, how much would you feel (upset/angry/ hate)? all scenarios were developed based on previous data from research regarding hate in human relationships (aumer et al., 2015; aumer et al., 2016). all scenarios were first piloted with seven research lab members who were unfamiliar with the study. these seven scenarios were finalized a er 100% agreement. scenarios were answered using a 5 point likert scale with 1 “not at all” to 5 being “extremely”. the scenario regarding a ack was answered on a 1 to 5 scale with 1 being “very unlikely” and 5 being “very likely”. pet a itudes participants rated their a itudes towards pets using templer and colleagues (1981) pet a itudes scale (pas) modified. the modified version allows for those who have not owned a pet, but have in the past or would like to in the future, take the scale (templer et al., 2004). additionally, the pas measures a itudes as opposed to a achment or commitment to the pet. this 18 item measure has questions like: “i like house pets” and “i hate animals (reverse scored)” and uses a rating scale from 1 to 7 with 1 being “strongly disagree” and 7 being “strongly agree”. given that the pas can be used as a general scale (templer et al., 2004) we did not examine the factors of the pas separately in relation to our hypotheses. creative common license 4.0 – non commercial – share alike – attribution page 21 aumer, erickson, krizizke, jaksuwijitkorn, & åbb pet behaviour science katherine aumer, michael erickson, jack krizizke, marc jaksuwijitkorn, jennifer åbb pet behaviour science 2022, vol. 13, 16 31 doi:10.21071/pbs.vi13.13473 creative common license 4.0 – non commercial – share alike – attribution page 22 results descriptives for relationship with human all participants named someone they loved and cared about, 47% named a parent, 30% named a significant other, 8% named a friend, 7.5% named a sibling, 7% named a family member other than a parent or sibling (e.g., grandparent, uncle, aunt, etc), and 1% named other, specifically “myself”. for the purposes of our study, we excluded the answers from the person who marked “myself” as the person they loved and cared about. we did find significant relationships between age and equity in human relationships; older individuals reported being more underbenefi ed (r(125) = –0.193, p = 0.031), age and enjoyment with pets; older people reported less joy being with their pet (r(124) = –0.182, p = 0.043), and older individuals were less likely to report hating pets (r(122) = –0.179, p = 0.048) or hating humans (r(122) = ‑0.196, p = 0.030) when in a negative situation. we controlled for age when analyzing the variables of equity, enjoyment, and hate. descriptives for relationship with pet most participants reported a dog or cat as a pet they currently (72 dog, 21 cat) or previously owned (27 dog, 9 cat). eleven participants reported a different animal they either currently, wanted, or previously owned (3 rabbit, 3 fish, 1 bird, 1 guinea pig, 1 goat, 1 monkey, 1 gecko). however for the purposes of this paper, we report only the results from those who reported a dog or cat as a current or previously owned pet (n = 138). additionally, 58 participants (42%) had missing answers either in a measure or as part of the demographics. we include their answers and report the results with the degrees of freedom reflecting the sample that was used in the analysis. comparison of human and pet relationships love and hate due to technical issues with the survey, only 53 people answered the love and hate questions. all other questions were not impacted. we compared the love in both human and pet relationships using repeated measures anova. preliminary one‑way anova with post‑hoc analyses using bonferroni correction revealed no differences in love towards dogs or cats, however we did find differences in love depending on the type of human relationship with friends (m = 4.43, sd = 0.535) being loved less compared to parents (m = 4.90, sd = 0.309), siblings (m = 4.60, sd = 0.548), other family members (m = 5.00, sd = 0.000), and significant others (m = 4.89, sd = 0.323), f(4, 48) = 2.997, p = 0.028. in the repeated measures anova we used relationship type as a between subjects variable to account for these differences. we found no differences of love between human (m = 4.81, sd = 0.395) and pet (m = 4.70, sd = 0.607) relationships. we also found no differences in hate between dog or cat owners or human relationship types. the repeated measures anova did not reveal any significant differences in hate between human (m = 1.11, sd = 0.393) and pet (m = 1.08, sd = 0.277) relationships. see table 1 for summary. 2022 vol. 13 16 31 www.petbehaviourscience.org katherine aumer, michael erickson, jack krizizke, marc jaksuwijitkorn, jennifer åbb pet behaviour science 2022, vol. 13, 16 31 doi:10.21071/pbs.vi13.13473 creative common license 4.0 – non commercial – share alike – attribution page 23 time spent a preliminary one‑way anova with post‑hoc analyses using bonferroni correction revealed that participants reported spending most of their time with their significant others (m = 4.13, sd = 0.686) compared to parents (m = 3.27, sd = 0.936), siblings (m = 3.40, sd = 1.265), other family members (m = 3.33, sd = 1.00), and friends (m = 3.18, sd = 1.25), f(4, 125) = 5.826, p < 0.001. repeated measures anova with human relationship type as a between subjects variable, compared time spent between human (m = 3.50, sd = 0.986) and pet (m = 3.56, sd = 1.040) relationships and was not significant. see table 1 for summary. enjoyment a preliminary one‑way anova with post‑hoc analyses using bonferroni correction revealed that participants reported most enjoyment from their relationships with significant others (m = 4.63, sd = 0.540) and other family members (m = 4.67, sd = .500) and the least amount of enjoyment from their relationship with their parents (m = 4.18, sd = 0.725), f(4, 125) = 3.548, p = 0.009. repeated measures anova with human relationship type as a between subjects variable, compared enjoyment between human (m = 4.39, sd = 0.645) and pet (m = 4.33, sd = 0.912) relationships and was not significant. see table 1 for summary. emotions during negative event scenarios we asked participants to rate their upsetness, anger, and hate towards a human or pet across different negative event scenarios. we also asked participants to rate the likelihood of a acking a human or pet if that human or pet a acked them. four paired samples t‑tests using bonferonni correction to control for type 1 error revealed that all differences concerning reactions to negative events were significant between humans and pets. we averaged ratings of upsetness, anger, and hate for pets and humans across all the negative event scenarios and conducted three paired samples‑tests. we found that people reacted with less upsetness, t(125) = ‑15.209, p <.001, cohenʹs d = ‑1.355, 95% ci difference [‑1.36, ‑1.05], anger, t(124) = ‑14.979, p <.001, cohenʹs d = ‑1.34, 95% ci difference [‑1.32, ‑1.01], and hate, t(125) = ‑6.924, p <.001, cohenʹs d = ‑0.62, 95% ci difference table 1. average (m) ratings of love, hate, time spent, and enjoyment for humans vs. pets. there were no significant differences between human and pet relationships for love, hate, time spent, and enjoyment. there was a significant difference for equity, with people reporting being more overbenefi ed in their human relationships and underbenefi ed in their pet relationships (*p < 0.001). aumer, erickson, krizizke, jaksuwijitkorn, & åbb pet behaviour science katherine aumer, michael erickson, jack krizizke, marc jaksuwijitkorn, jennifer åbb pet behaviour science 2022, vol. 13, 16 31 doi:10.21071/pbs.vi13.13473 [‑0.498, ‑0.28], towards a pet than they would towards a human in similar hypothetical negative events. see figure 1 for details. additionally, participants were more likely to report a acking back, at a human than a pet, t(124) = 4.321, p <0.001, cohenʹs d = 0.387, 95% ci difference [0.299, 0.805]. creative common license 4.0 – non commercial – share alike – attribution page 24 figure 1. differences in emotions between human and pet relationships. given negative scenarios in a relationship, people report a higher average of being upset, angry at, and hate towards humans in these negative scenarios than towards their pets. all differences between human pets were statistically significant (*p < .001). testing hypotheses equity to test our first hypothesis that people will report more equity with human relationships than pet relationships, participants answered the global measure of equity scale (traupmann, peterson, utne, & hatfield, 1981; young & hatfield, 2009) from a ‑3 (greatly underbenefi ed) to +3 (greatly over‑benefited) scale, with scores around 0 being most equitable. a one‑way anova with post‑hoc analyses using bonferroni correction revealed that participants reported being overbenefi ed in their relationships with their parents compared to all other human relationships (m = +1.43, sd = 1.731), f(4, 125) = 9.522, p < 0.0001, partial η 2 = 0.234. data were normally distributed, but violated the assumption of homogeneity of variance (levene statistic(4, 125) = 5.275, p < 0.001). the welch statistic demonstrates that the main effect is still significant (welch (4, 26.362) = 9.403, p < 0.001). to directly test the differences between human and pet relationships in terms of equity (h1) we conducted a repeated measures anova comparing equity between human and pet relationships with human relationship type as a between subjects variable, revealed that people report being more over‑ benefi ed in their human relationships (m = +0.611, sd = 1.62, 95% ci [0.233, 1.194]) and under‑benefi ed in their pet relationships (m = –0.5238, sd = 1.72, 95% ci [ ‑1.194, ‑0.233]), f(1,121) = 8.634, p = 0.004, partial η 2 = 0.067. the interaction between equity and the human relationship type was significant, f(4,121) = 5.657 p < 0.001, partial η2 = 0.158. mauchlyʹs test of sphericity was 1.00. examining the 2022 vol. 13 16 31 www.petbehaviourscience.org katherine aumer, michael erickson, jack krizizke, marc jaksuwijitkorn, jennifer åbb pet behaviour science 2022, vol. 13, 16 31 doi:10.21071/pbs.vi13.13473 simple effects analysis revealed that the difference in equity between human and pet relationships was only significant when people were comparing the equity in their parent relationship with their equity in their pet relationship (mean difference = 2.034, se = 0.271, p < 0.001, 95% ci [1.498, 2.571]). h1 was partially supported. see table 1 for summary. pet a itudes to test our second, third, and fourth hypotheses, we used average pet a itudes and emotions. average a itudes about pets and pet ownership was positive (m = 5.88, sd = 0.72). pet a itudes significantly predicted residual change scores for love, with higher pet a itudes related to more love directed towards pets than humans, ᵯ� = 0.59, t(47) = 5.049, p < 0.001, and explained a significant proportion of variance in the difference in reported love, r2 = 0.352, f(1, 47) = 25.497, p < 0.001. similar results were also found for time (ᵯ� = 0.520, t(117) = 6.590, p < 0.001; r2 = 0.27, f(1, 117) = 43.428, p < 0.001) and enjoyment (ᵯ� = 0.635, t(117) = 8.902, p < 0.001; r2 = 0.40, f(1, 117) = 79.239, p < 0.001). as people’s pet a itudes increased, their love, enjoyment, and time spent with pets was greater than that for humans and therefore h2 was supported. when it came to hate, pet a itudes did not predict residual change scores in hate (p = 0.733), therefore h3 was not supported. in regards to peopleʹs responses to negative event scenarios, participantsʹ pet a itudes were good predictors of their differences in anger and hate, with increases in pet a itudes predicting more anger (ᵯ� = ‑0.270, t(116) = ‑3.018, p = 0.003; r2 = 0.07, f(1, 116) = 9.108, p = 0.003) and hate (ᵯ� = ‑0.287, t(114) = ‑3.20, p = 0.002; r2 = 0.08, f(1, 114) = 10.24, p = 0.002) towards human companions in negative scenarios. additionally, pet a itudes were good at predicting the likelihood of a acking back, with higher pet a itudes leading to a greater likelihood to a ack back at a human, but not a pet if the human companion was threatened (ᵯ� = ‑0.287, t(116) = ‑3.227, p = 0.002; r 2 = 0.08, f(1, 116) = 10.41, p = 0.002). however, pet a itudes were not good predictors of residual change scores for upsetness, therefore, h4 was only partially supported. discussion the growing trends towards pet ownership may be impacted by several social and financial factors. whether people prefer pets over human companionship can also be impacted by several personal and social factors. in our study, we found some insights into how human‑pet and human‑human relationships are different as well as found support for most of our hypotheses. overall, people report having quality human and pet relationships, especially in regards to positive feelings like love, time spent, and enjoyment. however, more positive pet a itudes significantly predicted more positive feelings (i.e., love, time, and enjoyment) for pets than for humans. not supporting our hypothesis, pet a itudes did not predict differences in overall negative emotions, specifically hate. when it came to hypothetical negative events, participants seemed much more likely to react with anger and hate towards a human being than a pet if they had positive pet a itudes. again, this may be evidence that pets are o en seen as almost “child‑like” or dependent upon their human companions and such a negative emotional response may not be appropriate for some one or some creative common license 4.0 – non commercial – share alike – attribution page 25 aumer, erickson, krizizke, jaksuwijitkorn, & åbb pet behaviour science katherine aumer, michael erickson, jack krizizke, marc jaksuwijitkorn, jennifer åbb pet behaviour science 2022, vol. 13, 16 31 doi:10.21071/pbs.vi13.13473 creative common license 4.0 – non commercial – share alike – attribution page 26 animal in that role (franklin 1999; serpell 1996; serpell, 2003). while people who serve as parents, significant others, other family members, or friends may be viewed as acceptable targets of anger and hate given their independence and maturity. this willingness to provide preferential treatment to pets is further supported by the report that most people would not a ack their loved pet if the pet a acked them, but would be more likely to with a loved human who had a acked them. this study provides more supporting evidence to polls that find some preference for pet relationships compared to human relationships (haaland, 2019) and scholarly work (e.g., basten, 2009; belk, 1996; veevers, 1985; walsh, 2009) that pet relationships can serve a variety of functions, especially that of a surrogate for a achment love. additional evidence that pets are seen on a similar level with that of a child is the differences in equity scores. people reported being slightly more underbenefi ed with their pets than in their human relationships and this difference in equity was largely carried by the comparison of parent‑ child relationships to human‑pet relationships. people reported being significantly under‑benefited in their relationship with their pet, especially when compared to their relationship with a parent, bolstering evidence that pets do have sufficient costs along with benefits (vale & vale, 2009). importantly, differences in relationship quality between pets and humans was predicted by the explicit reports of one’s a itudes about pets in general. a itudes are general evaluations of an object (eagly & chaiken, 1998) and in this study, positive a itudes about pets were positively related to preferential treatment of pets over humans. those who reported more love, time, and enjoyment with their pets than their human relationship had more positive pet a itudes. similarly, those who were more likely to respond with anger, hate, and a ack back in negative situations towards their loved humans rather than their pets, also had more positive pet a itudes. although people were likely to report being as upset about a negative event, whether it involved a human or a pet, people were more likely to react with anger and hate towards a person than they were a pet. the impact of oneʹs a itudes about pets may not only impact the relationship they have with their pet, but with the humans in their lives. preferences for certain types of people, or in this case species, may impact how we relate to other potential relationship partners. this study provides some evidence that pet a itudes may be a good predictor of differences in the quality of relationships one may have between pets/animal companions compared to their human companions. for the ordinary pet owner, pets can provide companionship in ways a human companion may not be able to. specifically, pet relationships may serve as a surrogate to help fulfill the need to belong and a achment love that are not readily accessible from human companionship. for those that, for a variety of reasons, are not able to access or maintain human relationships and have positive pet a itudes, a pet may provide not just an acceptable outlet, but a mental and physical health benefit. finally, these results suggest that relationships, despite how costly or inequitable they may be, can provide significant positive relationship outcomes if the companion is viewed or framed not as a human, but as a companion requiring oneʹs presence to survive. 2022 vol. 13 16 31 www.petbehaviourscience.org katherine aumer, michael erickson, jack krizizke, marc jaksuwijitkorn, jennifer åbb pet behaviour science 2022, vol. 13, 16 31 doi:10.21071/pbs.vi13.13473 creative common license 4.0 – non commercial – share alike – attribution page 27 limitations the negative event scenarios are only an estimation of behavior. observation under a variety of situations may provide a be er indication of actual behavior. future research could use observation or informant reports to confirm or provide more insight into these results as well as the perspective of the pet. in addition to observation of behavior, future research may want to explore relationships with other pets besides dogs and cats. although dogs and cats are the majority species for pet owners in the u.s. it would be important to see if these results carry over to relationships with other pet species (strochak et al., 2018). we also did not measure a achment styles with pets. people’s a achments may also moderate or mediate the relationship between pet a itudes and relationship differences observed between human‑pet and human‑human relationships (kurdek, 2008; zilcha‑mano, mikulincer, & shaver 2011). no measures regarding human a itudes or a itudes about human relationships were administered. future research may want to consider creating and incorporating such measures. additionally, we did not incorporate any measures that would help be er understand the pets perspective. other researchers (serpell, 2003) have noted this drawback in other research as well and although we agree this is an important element in the research of pet and human relations, we were specifically interested in the human perspective of preference of pets and therefore did not incorporate any pet‑perspective measures. pet a itudes seem to be positively related to positive feelings and negatively related to negative responses towards pets. although pet a itudes accounted for a significant portion of variance in these situations, there are likely many other factors that may go into the inequitable treatment of human and pet relations. in this study, we asked participants to name a human and a pet they love and care about. we did not ask them to name their favorite human or pet or most ideal human or pet. we assumed the human or pet named by participants was one who received a significant, although possibly not similar amount of love and care. given that we wanted to compare differences in treatment towards humans and pets, we expected some variance. future research may want to try to maintain a certain standard of comparison between the human and pet named by the participant. finally, we did not measure the a itude towards human relationships. measuring this may also serve as an important indicator of treatment of human companions and pets. conclusion overall, people tend to target more negative emotions at other humans than animals in negative event scenarios. however, having positive pet a itudes can predict significant differences between pet and human treatment. a person’s pet a itudes may reflect more than just how they feel about their pet, but also indicate the dynamics of their human‑human relationships. as pet a itudes became more positive the report of loving a pet, spending more time with a pet, and enjoying their pet more than their loved one increased. also, positive pet a itudes were indicative of more negative reactions, specifically anger and hate towards humans compared to pets in the same negative scenarios. although we did not assess preference for human or pet relationships, pet a itudes may be a aumer, erickson, krizizke, jaksuwijitkorn, & åbb pet behaviour science katherine aumer, michael erickson, jack krizizke, marc jaksuwijitkorn, jennifer åbb pet behaviour science 2022, vol. 13, 16 31 doi:10.21071/pbs.vi13.13473 creative common license 4.0 – non commercial – share alike – attribution page 28 useful measurement to assess this preference. given that pet ownership is increasing and the use of an animal companion is o en recommended to help with physical and psychological health (smith, 2012), assessing pet a itudes may be a good indicator to see if animal companionship would be beneficial. acknowledgements mahalo (thank you) to amanda daniels, jun sato, briana staples, william hagblom, dylan light, and kristin gray who all tested the survey and engaged in conversations around people and their relationship to pets. references american pet products manufacturers’ association (appma). 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(2011). an a achment perspective on human–pet relationships: conceptualization and assessment of pet a achment orientations. journal of research in personality, 45(4), 345‑357. creative common license 4.0 – non commercial – share alike – attribution page 31 aumer, erickson, krizizke, jaksuwijitkorn, & åbb pet behaviour science katherine aumer, michael erickson, jack krizizke, marc jaksuwijitkorn, jennifer åbb pet behaviour science 2022, vol. 13, 16 31 doi:10.21071/pbs.vi13.13473 food preference predicts speed of approach on a runway task by dogs food preference predicts speed of approach on a runway task by dogs kristie e. cameron1*, jane de garnham1, kristeen jensen1 & lewis a. bizo2 pet behaviour science | 2019, vol.8, 1-10 doi: 10.21071/pbs.v0i8.11179 kristie e. cameron, jane de garnham, kristeen jensen & lewis a. bizo 1 unitec institute of technology, auckland, new zealand * environmental and animal sciences pathway unitec – institute of technology carrington road, auckland new zealand tel: +64 9 8154321 ext 8578 2 university of new england, armidale, australia research paper * email: kcameron@unitec.ac.nz keywords: dog, food preference, reinforcer assessment, raw food, response latency; runway highlights • high-value foods are reported to function as more effective reinforcers than low-value foods. • dogs do not value staple foods highly. • dogs move faster in a runway ask for highly preferred foods compared to staple or lowly preferred foods. • dog owners are encouraged to conduct preference assessments to identify their dog’s most preferred food for use as a reinforcer during training. page 1creative commons license 4.0 – non commercial – share alike – attribution 2019 | vol. 8 | 1 10 abstract the effective and quick assessment of food preference is important when attempting to identify foods that might function as effective reinforcers in dogs. in the current experiment, more highly preferred foods were expected to be associated with faster approaches in a subsequent runway task. eight dogs experienced combinations of two of six types of raw food in a paired preference assessment. these included the dog’s staple diet, to identify a rank order of preference for the foods. a different raw food was offered as the staple in two preference tests. in the runway task, the dogs were required to walk five metres to obtain a small amount of their most preferred, least preferred or staple foods and latency of approach to the foods was recorded. the results showed that the staple foods were not preferred as highly as the other foods and that each dog displayed unique and stable preferences for the different foods. the approach latencies were faster for their most-preferred food compared to their least preferred and the staple foods. the use of a runway to assess reinforcer effectiveness combined an effortful behaviour to obtain food while also requiring the dogs to make a choice, thus precluding the need for more complicated and time-consuming methods of preference assessment. the application of this method for fast and effective identification of preferred reinforcers is currently being investigating further to inform pet owners and behavioural scientists better about simple methods that they might use to identify highly preferred foods for use as reinforcers in training and behavioural testing. http://www.petbehaviourscience.org/ introduction to promote the welfare of domestic pets, it is important that pet owners employ positive training techniques. an example of a positive training technique is the delivery of a reward that is contingent on an animal producing the appropriate behaviour. delivery of a reward should increase the likelihood an animal will repeat that behaviour in the future (e.g., hiby, rooney and bradshaw 2004; haverbeke, laporte, depiereux, giffroy and diederich 2008;); a technique used in behavioural science where the occurrence of behaviour is measured in relation to the delivery of reinforcement, usually food, according to a schedule of reinforcement. in addition, rewarding desired behaviour is reported to strengthen the human-animal bond (deldalle and gaunet 2014; payne, bennett and mcgreevy 2015), decrease animal stress during training (deldalle and gaunet 2014) and mitigate the development of problematic behaviour which can result in relinquishment of the dog by the owner if it persists (blackwell, twells, seawright and casey 2008). to successfully implement positive reinforcement, one must use a functional reinforcer; this means that the reinforcer is highly valued by the animal (gaalema, perdue and kelling 2011; vicars, miguel and sobie 2014). conducting systematic preference assessments in humans is a method of identifying food and leisure items to reward desired behaviour (deleon and iwata 1996; fisher et al. 1992). more recently the same technology has been used by animal keepers to provide preferred food and enrichment to animals such as with: horses (equus caballus; elia, erb and houpt 2010); orangutans (pongo spp.; clay, bloomsmith, marr and maple 2009); cats (felis silvestris catus; vitale shreve, mehrkam and udell 2017); giant pandas (ailuropoda melanoleuca) and african elephants (loxodonta africana; gaalema et al. 2011), all of which found idiosyncratic differences between individuals of each species. behavioural scientists also use these tests to identify preferred foods to supply as reinforcers in operant experiments with various animals including brushtail possums (trichosurus vulpecula; cameron, bizo and starkey 2013) and hens (gallus gallus; sumpter, foster and temple 2002) and to examine preferred or aversive environmental components such as ammonia concentration by sheep (ovis aries; phillips, pines and muller 2012). one particular instance where this is particularly important is when animals are on a restricted diet such as in a laboratory environment where the motivation to perform for food is often a critical component of the experimental method (cameron, bizo and starkey 2015) or a veterinarian has suggested a specialised diet. there are populations of pet dogs that are fed raw food diets for various reasons (ackerman 2016). given these dogs are unable to consume commercial treats, it is necessary to be able to identify foods they can eat, that will be highly valued, and that would function as a reward in dog training and operant experiments to motivate and reinforce the occurrence of target behaviour for this population. for this experiment, raw food fed dogs were chosen as subjects for this study because it is recommended by commercial sellers of raw food that dogs should not eat store-bought treats or high-value human food as it decreases their gut-acidity and results in the dog having digestive issues and gastrointestinal upset (thompson 2016). instead, owners have reported using a particular ‘flavour’ of their dogs staple raw food such as rabbit, horse or veal mixed with tripe in packets sold by retailers as a reinforcer or reward for their dog. it is unclear, however, given these dogs are restricted to raw food, whether using a dog’s normal diet would be as effective as a reinforcer during training as other novel foods. it is reported in numerous studies that a staple food, such as ‘dry dog biscuit’ is commonly of lower preference than other types of food such as sausage, cheese or ‘treats’ as indicated in preference and reinforcer assessments with dogs (e.g., thompson, riemer, ellis and burman 2016; riemer, ellis, thompson and burman 2018). others have identified a ‘novelty’ or ‘monotony effect’ in preference tests with dogs and cats where novel foods are preferred over a long-term staple diet (e.g., ferrell 1984; bradshaw 2006; vondran 2013). there is; however, a report of puppies preferring the diet on which they were weaned over a diet of novel foods (ferrell 1984). a well-researched method used for assessing preference with animals is the paired stimulus preference assessment (e.g., cameron et al. 2013; clay et al. 2009) as it requires little effort on the part of the page 2 www pet behaviour science org creative commons license 4.0 – non commercial – share alike – attribution 2019 | vol.8 | 1 10 researcher, does not involve large amounts of food and is relatively quick to administer (vicars et al. 2014; riemer et al. 2018;). it is also a valid and reliable method for assessing preference and involves pairs of food or items being systematically offered to the subject with their choice recorded (fisher et al. 1992) and is reliable over time (e.g., cameron et al. 2013). the use of operant manipulanda, where an animal can respond on a lever or a response key so that they can either work for or indicate a choice of a particular commodity is not a new technology. collier, hirsch and hamlin (1972) required rats to respond on increasing fixed ratio schedules to earn their entire food ration. this closed economy resulted in consistently high numbers of responses. the requirement to earn one’s food is clearly a great motivator. responding to simple schedules of reinforcement has been used to measure the value of reinforcers to an organism in single (e.g., bizo and killeen 1997; jarmolowicz and lattal 2010) and concurrent arrangements (e.g., sumpter et al. 2002) and the demand for commodities by requiring them to commit physical effort to obtain a particular item (e.g., hursh, madden, spiga, deleon and francisco 2013). tests designed to measure the effort put forth to gain a commodity have been conducted in a variety of animals such as possums (cameron et al. 2015; cameron, clarke, bizo and starkey 2016), horses (elia et al. 2010) and dogs (vicars et al. 2014). previous studies have reported that foods of low preference, including staple foods, produced lower rates of responding compared to foods of high value, for example to those with a higher sugar content, such as berries for possums (cameron et al. 2015) and high protein or fat coated treats for dogs (e.g., rashotte, foster and austin 1984; hewson-hughes et al. 2012). more recently, researchers have investigated methods of measuring captive animal’s ‘demand’ for commodities such as particular foods by pressing levers in possums (cameron et al. 2015; cameron et al. 2016), pressing keys in hens (e.g., foster, sumpter, temple, flevill and poling 2009), using a touch response (vicars et al. 2014) or runway movement for two particular commodities in dogs (riemer et al. 2018; thompson et al. 2016). a ‘work’ requirement has also been used to measure demand for a variety of commodities such as substrate in pigs (e.g., holm, jensen, pedersen and ladewig 2008) and hens (e.g., de jong, wolthuis-fillerup and van reenen 2007), and enclosure enrichment in various species kept in captivity such as lizards (e.g., januszczak et al. 2016), and rabbits (e.g., seaman, waran, mason and d’eath 2008). an animal indicating a need for a particular event or commodity, by committing physical effort in responding to obtain it, suggests that it should be provided to maintain the wellbeing of the animal (dawkins 1988; 2004). the same logic should hold for selecting a functional reinforcer for successful training; if an animal commits physical effort to obtain one type of food over another, it should be used as a reinforcer for training to be effective. preference and reinforcer assessments have been conducted with dogs to measure the palatability of commercially available dog food where dogs display a tendency to choose a novel food option over their staple diet (e.g., vondran 2013). dogs have also been observed to forego a low-value food or a small quantity of food for a high-value or larger amount of food (leonardi, vick and dufour 2012), indicating an ability to discriminate between larger and smaller amounts of food (e.g., mcguire, bizo, mcbride and kocek, 2018). dogs showed a preference for dried meat, cheese or treat-type foods over mundane dog food and biscuits. the dogs did not respond for the low-value food but responded to higher response requirements and for longer to obtain the most-preferred food type when the dogs were required to perform a nose-touch response to a fixed object to obtain the mostor least-preferred food in a reinforcer assessment task (vicars et al. 2014). dogs reinforced with a high-value liver treat held the experimenter’s gaze for a longer duration than dogs either trained using dog pellets or those that had been reinforced previously with the high-value treat (bentosela, jakovcevic, elgier, mustaca and papini 2009). similarly, dogs spend more time interacting with an inaccessible high-value meat reinforcer than dry food in tests where the food was covered with a wire-netting cover (thompson et al. 2016). when dogs are exposed to choice tests of foods of differing qualities and quantities they tended to select the larger amount and more highly valued foods more often (riemer et al. 2018). the authors concluded that a more highly valued page 3 food preference on a runway task by dogs creative commons license 4.0 – non commercial – share alike – attribution cameron, garnham, jensen & bizo https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science food is likely to be a more effective reinforcer, and that is not likely to include staple food. the current study aimed to assess the appropriateness and practicality of a preference assessment method using a combined effortful and choice procedure to assess preferences in dogs. this involved identifying a rank order of preference for six flavours of raw food using a paired stimulus preference assessment and then assess if that rank order predicted rates of responding for those foods on a simple schedule of reinforcement. the reinforcer assessment required dogs to walk down a 5-m runway to obtain their staple, mostor least-preferred foods. we predicted that dogs would approach their most-preferred food faster compared to a staple or least-preferred food. method subjects eight domestic dogs of various breeds participated in the experiment (table 1), and owners gave their permission before testing. the university of auckland ethics committee approved this research (approval number 001769). table 1. subject information; name, sex, breed and life stage of each dog. ‘food’ represents the amount of food offered per trial name (sex) breed life stage food (g) bradley (m) australian shepard adult 5 miika (f) alaskan malamute adult 5 indy (f) rhodesian ridgeback x adult 5 moe (m) mixed breed young adult 5 max (m) border collie adult 5 poppy (f) miniature schnauzer adult 3 rex (m) maltese shih tzu adult 3 runty (f) staffordshire terrier x young adult 3 skye (f) german shepard adult 5 apparatus trials were conducted indoors in the dog’s home or in a private room at the local doggy-daycare facility where the dog was familiar. a ‘virtual runway’ was created with a straight space approximately 2-3m long where two plates with a food sample on each marked the end of the runway. before the first paired-stimulus preference assessment, owners were instructed to feed their dog their normal ration of a specific raw food type for each meal for the three days before testing. dogs were offered different food types of the same branded raw food made from a “natural, preservative free, species-appropriate raw food diet” (gourmet petfood kitchen limited, 2015). before the first test, the dogs received ‘rabbit,’ and before the second test conducted two weeks later, they received ‘horse.’ the other foods offered were chicken, tripe, duck and lamb. to calculate the amount of food offered each dog would receive in a trial, the dog’s normal morning ration of food was divided by the number of trials conducted in a session (approximately 36). a food sample of this amount of food type was then offered to the dog on each trial during the assessment (see table 1 for the amount of food each dog received). procedure the paired preference assessment consisted of 30 trials where food presentation was counter-balanced across sides each food was paired with every other food ̶ and offered on the left and right plate in separate trials. the food pairs were presented in a pseudorandom order such that the same foods were not offered on successive trials. the experimenter would prepare the test foods according to the pre-determined order of presentation and replace the plate as the dog was walked to the start point. plates were washed thoroughly between dogs. at the beginning of each trial, the dog would be on a loose lead in a ‘sit’ position at the beginning of the route. the owner would instruct the dog to move and then walk behind the dog to avoid developing an owner-induced side bias to the food samples. initially, page 4 www pet behaviour science org creative commons license 4.0 – non commercial – share alike – attribution 2019 | vol.8 | 1 10 each food was presented singly on either the left or right side to familiarise the dog with the procedure. once a food was selected, operationalised as the dog picking the food up in their mouth and not expelling it, the owner would allow the dog to eat the food. the owner then led the dog in the same direction as the dog’s choice (left or right) in a circular motion back to the starting point to await the next trial. reinforcer assessment the dogs were required to move from a sitting position down the ‘runway’ to a point 5-m distant for a sample of either their most, least preferred or staple foods. owners would hold their dogs in place by the collar then release the dog to move toward the food. the latency from the ‘start’ to the consumption of food was measured. the owner would then attach a lead and return the dog to the start position. to ensure the reliability of the impact of a dog’s demand for an individual food item the dog was given five separate opportunities to obtain each food in an abcd repeated measures design; (a = most-preferred food for 5 trials, b = least-preferred food for 5 trials, c = rabbit (or lamb for indy) as the staple food for 5 trials, and d = horse as the staple food for 5 trials). results all food types were selected at least once by all dogs in the paired stimulus preference assessments. table 2 shows the rank order of preference for each dog and the proportion of trials (out of 20) when each food was chosen across the two sessions. wilcoxon signs ranks tests revealed no differences between the rank orders of food for the first preference test when rabbit was the staple food, and the second test when horse was the staple food for all of the dogs [bradley w = -0.41, p = 0.684; indy w = -0.27, p = 0.786; max w = 0.00, p = 1.00; miika w = 0.00, p = 1.00, moe w = 0.00, p = 1.00; poppy w = -0.41, p = 0.686; rex w = -0.11, p = 0.916; runty w = 0.00, p = 1.00; skye w = 0.00, p = 1.000]. overall, lamb, duck, and tripe featured as the most-preferred food for two each of the nine dogs, with the remaining dogs indicating their most-preferred food was either chicken, rabbit or horse. preference order dog 1st 2nd 3rd 4th 5th 6th bradley c (0.60) d (0.60) l (0.60) t (0.45) r (0.40) h (0.35) indy l (0.70) d (0.60) r (0.55) c (0.40) h (0.40) t (0.35) max l (0.75) d (0.70) t (0.50) r (0.45) c (0.30) h (0.30) miika d (0.75) l (0.60) t (0.55) h (0.50) c (0.30) r (0.30) moe d (0.85) c (0.60) l (0.60) h (0.40) t (0.35) r (0.20) poppy r (0.75) c (0.60) d (0.50) l (0.45) h (0.35) t (0.35) rex t (0.75) h (0.65) d (0.45) l (0.45) c (0.35) r (0.35) runty h (0.75) t (0.60) r (0.50) c (0.40) d (0.40) l (0.35) skye t (0.75) l (0.60) d (0.60) r (0.50) c (0.40) h (0.15) table 2. relative preference for each food type (c chicken; d duck; l lamb; t tripe; r rabbit; h horse) for each dog summed across two sessions. the proportion of trials when each food was chosen is given in parentheses. the average proportion of trials when each food was chosen was compared across dogs to assess whether preference was affected by the quality of staple food (figure 1). the data for indy was not included in the analysis as she received lamb as a staple food instead of rabbit. overall duck was preferred in significantly more trials than chicken when rabbit [z = -2.3, p = 0.050] and horse were offered [z = -2.17, p = 0.038]. preference for the each of the staple foods was not significantly higher when that food was available as the staple food; for rabbit [z = -1.83, p = 0.068] and for horse [z = -0.99, p = 0.322]. the reinforcer assessment required each dog to move 5m to earn a sample of food. the samples were the mostand least-preferred foods and the staple foods for each dog. as there were five trials per food type, the first trial was omitted in the analysis (figure 2). a friedman statistical analysis revealed significant differences between the average latencies to travel 5-m to obtain either the mostand least-preferred, and the two staple foods [χ2 (3) = 36.98, p < 0.001, w = 0.39]. page 5 food preference on a runway task by dogs creative commons license 4.0 – non commercial – share alike – attribution cameron, garnham, jensen & bizo https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science data for one dog (runty) was omitted from the analysis as their most-preferred food was not tested during the reinforcer assessment. pairwise comparisons revealed significantly faster average latencies to receive the most-preferred food compared to both staple foods [all p’s < 0.001], and the least-preferred food [p < 0.001]. latencies to obtain the least-preferred foods were significantly slower than for the rabbit staple [p = 0.005] and horse staple [p = 0.024]. there were no significant differences in latency to obtain staple foods [p = 0.270]. the absence of a difference in latency means that when the dogs are required to expend effort to obtain a reinforcer, they will move faster for a food of higher value compared to their staple diet or least-preferred food. discussion in the current study, a paired stimulus preference assessment was used to determine a rank order of foods preferred by dogs fed a specialised ‘raw’ diet and to measure the effect of staple foods on preference. there were two factors of interest, firstly, whether the rank order of preference was stable over time and if the preference for the staple foods was affected by the provision of these foods (as the normal diet) in the three days before testing. the results indicate a high degree of idiosyncrasy in preference similar to that identified previously in dogs (e.g., vicars, et al. 2014) and other species such as possums (cameron et al. 2013). the data also showed that some dogs would cover a short distance faster to obtain their mostpreferred food compared to a lowly-preferred or staple food similar to previous works where dogs needed to ‘move’ to gain a reinforcer (e.g., riemer et al. 2018; thompson et al. 2016). to measure the effect of the staple, most and least preferred foods on behaviour, we used a task combining effort and choice. the dogs were required to cover a 5-m distance to obtain a reinforcer. this method produced significant differences in latencies for five dogs when offered the reinforcer options and provides an ‘unconditioned’ utility for measuring effort to obtain a reinforcer in shelter, young or minimally trained dogs. the utility of quick and reliable preference assessments is of considerable applied interest. a variety of methods and techniques have been developed that vary in complexity and ease of use. these extend from simple single stimulus presentations that are useful for palatability tests to paired stimulus and multiple stimulus presentations methods that provide rank orders of preference for foods (for examples see cameron et al. 2013, fisher et al. 1992 and page 6 www pet behaviour science org creative commons license 4.0 – non commercial – share alike – attribution 2019 | vol.8 | 1 10 test foods chicken duck tripe lamb horse rabbit a v e ra g e p ro p o rt io n o f t ri a ls 0.0 0.2 0.4 0.6 0.8 1.0 rabbit staple horse staple figure 1. proportion of trials when each test food was selected by each dog. the darker columns represent the proportion of trials when rabbit was the staple food; the light grey columns represent the proportion of trials when horse was the staple food. the data for indy was not included in the analysis. error bars are the standard error of the mean. sumpter et al. 2002). the reinforcer assessment for each dog consisted of blocks of five trials of the most-, least preferred and the two staple foods, presented in that order to each dog. latencies were longer latencies in the final trials of the session which one could attribute to an order effect, especially as they were receiving their staple food. the data in figure 2, however, shows that dogs moved with similar latencies for both staple foods across the penultimate (rabbit staple) and final set (horse staple) of five trials. in future experiments, the order of presentation for the staple, most and least-preferred foods will be randomized across dogs and repeated sessions. furthermore, as the task required to obtain the food was not behaviourally different from that of the paired stimulus preference assessment that all dogs had experienced in earlier sessions, the ‘novelty’ of the task is unlikely to have caused the dogs to move faster to gain the food for any reason other than whether the food was of highor low-value to the dog. use of a runway in previous experiments with dogs have functioned more so as a reinforcer assessment and have measured either latency to approach (riemer et al. 2018) or interactions with inaccessible food (thompson et al. 2016). they did not provide different choices for food over trials as one might in a preference assessment. in the current experiment, the design of the preference assessment combined an effortful behaviour to obtain food, while also choosing competing foods, effectively precluding the need for a reinforcer assessment; as the results of the assessments corroborated the identification and effectiveness of the reinforcers. the requirement to move down the runway functioned as a fixed ratio schedule of reinforcement and if latency to approach the food was measured concurrently with choice one could simultaneously identify the relative demand of two commodities in dogs. future research will attempt to determine a reliable and easy to administer preference-assessment methodology that can be used by dog owners to easily and reliably assess their dog’s preferred reinforcer. also, a comparison of different preference assessment methods such as the paired stimulus preference assessment and multiple stimuli without replacement methods (mswo; deleon and iwata 1996) with the results of a reinforcer assessment. by presenting a dog with an effortful task that results in a reinforcer of their choice one would predict that an animal would select the foods of highest-value before they are willing to repeat the work required to choose the next most preferred item. such a technology would hopefully improve the likelihood that chosen rewards would function as reinforcers and page 7 food preference on a runway task by dogs creative commons license 4.0 – non commercial – share alike – attribution cameron, garnham, jensen & bizo food type most least staple (r) staple (h) a ve ra g e l a te n cy ( se co n d s) 0 1 2 3 4 5 6 figure 2. average latency (seconds) to complete a 5 m distance across dogs for the mostand least preferred and staple food (r = rabbit; h = horse). the food types are shown in order of presentation to the dogs with the final four trials used for each block of trials. error bars are the standard error of the mean. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science improve training outcomes. in conclusion, this study has identified that a high value ‘flavour’ could be withheld and used as a reinforcer for training dogs on a restricted diet. the present study has demonstrated the utility of simple reinforcer preference assessments with dogs and builds on a previous body of research on humans and animals that attempts to find practical and cost-effective methods for assessing reinforcer preference. our method was practical and reliable and will be a useful tool for pet owners and behavioural scientists in determining a high-value food for reinforcing animals for the desired behaviour in training and behavioural testing. acknowledgements portions of these data were also reported at the new zealand companion animal council conference in auckland, new zealand 2018. the authors would like to thank the dog owners and dogs for participating in the experiment. this research was conducted following the relevant ethics guidelines and approved by the animal ethics committee at the university of auckland where this research was conducted. the first author can be contacted at unitec institute of technology; kcameron@unitec.ac.nz. references ackerman, n. 2016. evidence surrounding the feeding of natural/raw diets to dogs and cats. the veterinary nurse 7: 5–8. bentosela, m., jakovcevic, a., elgier, a. m., mustaca, a. e., and papini, m. r. 2009. incentive contrast in domestic dogs (canis familiaris). journal of comparative psychology 123: 125–130. bizo, l. a., and killeen, p. r. 1997. models of ratio schedule performance. journal of experimental psychology: animal behavior processes 23: 351–367. blackwell, e. j., twells, c., seawright, a., and casey, r. a. 2008. the relationship between training methods and the occurrence of behavior problems, as reported by owners, in 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(2018). discrimination of food amounts by the domestic dog (canis familiaris). international journal of comparative psychology, 31. page 9 food preference on a runway task by dogs creative commons license 4.0 – non commercial – share alike – attribution cameron, garnham, jensen & bizo https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science payne, e., bennett, p. c., and mcgreevy, p. d. 2015. current perspectives on attachment and bonding in the dog-human dyad. psychology research and behavior management 8: 71–79. phillips, c. j., pines, m. k., and muller, t. 2012. the avoidance of ammonia by sheep. journal of veterinary behavior: clinical applications and research 7: 43–48. rashotte, m. e., foster, d. f., and austin, t. 1984. twopan and operant lever-press tests of dogs’ preference for various foods. neuroscience and biobehavioral reviews 8: 231–237. riemer, s., ellis, s. l., thompson, h., and burman, o. h. 2018. reinforcer effectiveness in dogs—the influence of quantity and quality. applied animal behaviour science 206: 87-93. seaman, s. c., waran, n. k., mason, g., and d’eath, r. b. 2008. animal economics: assessing the motivation of female laboratory rabbits to reach a platform, social contact and food. animal behaviour 75: 31–42. sumpter, c. e., foster, t. m., and temple, w. 2002. assessing animals’ preferences: concurrent schedules of reinforcement. international journal of comparative psychology 15: 107-126. thompson, h., riemer, s., ellis, s., and burman, o. 2016. behaviour directed towards inaccessible food predicts consumption – a novel food preference test. animal cognition 178: 111–117. thompson, l. 2016. raw essentials faq. retrieved from http://www.rawessentials.co.nz/site/webpages/general/ gastric-acidity vicars, s. m., miguel, c. f., and sobie, j. l. 2014. assessing preference and reinforcer effectiveness in dogs. behavioural processes 103: 75–83. vitale shreve, k. r., mehrkam, l. r., and udell, m. a. r. 2017. social interaction, food, scent or toys? a formal assessment of domestic pet and shelter cat (felis silvestris catus) preferences. behavioural processes, 141: 322–328. vondran, j. c. 2013. a two pan feeding trial with companion dogs: considerations for future testing. unpublished master’s thesis, kansas state university, kansas, usa. page 10 www pet behaviour science org creative commons license 4.0 – non commercial – share alike – attribution 2019 | vol.8 | 1 10 influence of dog presence on the tolerance and evaluation of aversive stimulation dogs in the criminal justice system: consideration of facility and therapy dogs elizabeth spruin* and katarina mozova highlights • there are important differences between therapy and facility dogs and their utility pet behaviour science | 2018, vol.5, 1 – 12 doi: 10.21071/pbs.v0i5.10084 elizabeth spruin* and katarina mozova canterbury christ church university review * email: liz.spruin@canterbury.ac.uk uk keywords: criminal justice, facility dog, therapy dog, victim, witness • dogs have a place in the criminal justice system • anecdotal evidence portrays the use of dogs as a beneficial for victims, witnesses and defendants. • research surrounding the use of dogs in the legal setting is lacking introduction whilst the role dogs’ play in human society has had a long and complex history, the impact that these animals have had on our society is undeniable. dogs have been shown to enhance human wellbeing, through simple page 1creative commons license 4.0 – non commercial – share alike – attribution 2018 | vol. 5 | 1 12 abstract it is known that the criminal justice process is most often perceived as a negative experience by victims, witnesses, as well as defendants. whilst measures have been put into place across the globe to improve their experiences, there is still much more which needs to be done, especially as the process can involve secondary victimisation of those participating in it and prolonged trauma. the current opinion piece centres on the use of trained dogs to help the experiences of criminal justice system users during active cases. whilst this practice is mostly used in north america, hints at bringing varying types of dogs into the criminal justice system are visible elsewhere, too. with the criminal justice users in mind, it is key to establish, from the offset, the positives of such service, but also be very aware of its limitations and challenges, in order for the service delivering what it aims without causing a disruption to the criminal justice process or its users. this piece provides a theoretical and practical analysis of topics surrounding the use of specially trained dogs to support criminal justice system users with the view of highlighting our lack of knowledge on the topic and practical challenges of this service. http://www.petbehaviourscience.org/ pet ownership or active intervention (friedmann 1995; fine and mackintosh 2016). they have been used in a variety of settings (e.g. schools, nursing homes, etc.) and for centuries, dogs have also been utilised within law enforcement where they are specially trained to fulfil a specific role within a variety of areas, from searching for drugs (lorenzo et al. 2003) and explosives (gazit and terkel, 2003), to locating suspects or finding evidence and missing people (browne, et al. 2006). only recently, the use of dogs within the criminal justice system to specifically help its users’ wellbeing (victims, witnesses, defendants) has been explored, originating in the usa through pioneering work of the courthouse dogs foundation, the only not-for-profit organisation advocating for the use of specially trained facility dogs to be utilised to help vulnerable individuals throughout the criminal justice system process (courthouse dogs foundation, 2018). this is due to overwhelming evidence highlighting the negative associations criminal justice system users create with their participation (e.g beckett and warrington 2015), the need to better care for them so they can provide their best evidence (e.g. o’mahony, et al. 2016), and the potential of the process re-victimising individuals (i.e. secondary victimisation; e.g. jordon 2013). consequently, a multitude of areas for consideration, not necessarily present in other settings, arise, as dogs in this setting are utilised during active cases which is associated with legal challenges. it must be noted that legal systems across the world differ and this piece is not an attempt at highlighting legal issues specific to certain countries; however, some common challenges are considered here which are likely to be visible in a variety of legal systems. further, academic literature is vastly lacking in certain areas and so other available literature was considered – it was attempted to only utilise unbiased, objective literature (e.g. national regulatory bodies); where this was not possible, information was provided as an example only. dog typology: therapy and assistance dogs when discussing or researching the effects that dogs can have on human wellbeing, therapy dogs or assistance dogs are utilised (e.g. berry, et al. 2013); however, differences between these categories are often neglected; an effect of the presence of, or interaction with dogs generally, tend to be considered (e.g. hoffmann, et al. 2009; lundqvist, et al. 2017), rather than their effect based on different training and type of intervention. these two categories are vastly different in their behaviours, training, or demeanour and such differences must be appreciated especially when utilising dogs within a legal setting where the ramifications of how dogs are utilised are important. first, brief background relating to these types of needs to be understood. since the turn of the 20th century, specific dog breeds have been purposefully bred and specially trained as assistance dogs to aid people with disabilities (fossum 2013). different breeds of dogs are, historically and contemporarily, associated with different behaviours (such as playfulness, sociability, aggressiveness; svatberg 2006); further breeding of specific breeds to fit with a job role ensures trainability and predictability of dogs to fulfil a role appropriately (weiss & greenberg 1997; wilsson and sundgren 1997). one such role is that of assistance dogs which fall into two broad and distinctive categories: service dogs and facility dogs. whilst both types of assistance dogs receive extensive training before becoming working dogs, their roles differ. in general, service dogs are trained to do work or perform tasks for people with disabilities, such as visual and hearing impartments, mental illnesses and immobility needs, therefore performing duties such as guiding people who are blind, alerting people who are deaf, pulling a wheelchair, alerting and protecting a person who is having a seizure, or reminding a person with mental illness to take prescribed medications (camp 2001; cavalli et al. 2017; huss 2017). a facility dog is another type of assistance dog that works alongside a professional in a service capacity to assist multiple people with different needs (tedeschi et al. 2010; himot, et al. 2017). in this capacity, both the facility dog and working professional are specially trained. therefore, where a service dog works for one specific person and is trained for their needs, a facility dog works with a professional within an institution and provides services to a variety of individuals. typical situations in which a facility dog would be utilised would be in educational settings to help facilitate interaction with students, or in healthcare environments to help patients with symptom management (e.g. bradley and maldonado 2013; krause-parello et al. 2016; lutwack-bloom et al. 2008). page 2 www pet behaviour science org creative commons license 4.0 – non commercial – share alike – attribution 2018 | vol.5 | 1 12 on top of task-oriented benefits, dogs have also been shown to affect human wellbeing in a variety of ways, such as through decreasing anxiety and pain (barker, et al. 2015), attenuating cortisol and heart rate (polheber and matchock 2014), or promoting simple feelings of calmness and happiness (carew-lyons 2016). as a result, and due to the wide-ranging possibilities of providing such assistance to people, the use of therapy dogs has been introduced in a number of contexts (e.g. nursing homes, majic et al. 2013; prisons, zimmer 2014; or schools and universities, barker et al. 2016) to provide affect and comfort to people, showing positive results. therapy dogs, however, are vastly different to assistance dogs. they are not trained to assist individuals, but rather, they are personal pets. guidelines as to what criteria a therapy dog needs to fulfil vary across countries. usually, they are evaluated based only on their temperament (i.e. how a dog reacts in new or unexpected situation) and have to pass a medical exam (grangen and kogan 2006). therapy dogs can undergo further training before being utilised (lafrance et al. 2007; obrusnikova et al. 2012); however, for their certification, this is not necessary and it is at the discretion of the institution which aims to include them in their work whether supplementary training is needed (connor and miller 2000; schoenfeld-tacher et al. 2017). generally, the selection criteria for therapy dogs to participate in a therapeutic relationship, their level of training or skill set are unsystematic across the world which has implication relating to their possible lack of predictability, sociability or controllability (cavalli et al. 2017; grangen and konan 2006; mongillo, et al. 2015). further, it has been found that the guidelines relating to how therapy dogs are certified are open to interpretation and so this assessment is not consistent (weiss, 2002). understanding the general background of these dogs, it should not come unexpectedly that as the important role that dogs play within human society continues to expand, it is crucial that best practice guidelines and standards are not only established, but also strictly adhered to. as each of these types of dogs have a very different training from the other, interchanging the terms and job roles can therefore be detrimental in diluting their value and creating issues of liability and legal issues. both types of dogs require accreditation from a recognised body (e.g. assistance dogs international; therapy dogs international) which set national or international standards relating to minimum level of training or skills; though variations in training still exist. further, not all countries are members of these organisations and so specific regulations relating to these need to be considered by professionals. more specifically, both facility and service dogs are intentionally raised and socialised from early puppyhood in order to effectively support individuals; facility dogs are professionally trained for two years by an internationally recognised assistance dog international (adi) program, which sets strict standards (cavalli et al. 2017; tedeschi, et al. 2010). facility dogs also work with a volunteer or professional who is also trained by an adi program. the facility dog, as well as the handler, have to be re-assessed regularly by the organisation. these dogs therefore work with a variety of people directly through their trained handler. both, service and facility dogs, are socialised throughout their training with people of different ages and backgrounds, other animals, and are not reactive to disturbing stimuli (crenshaw and stella 2015). in contrast to assistance dogs, as therapy dogs are personal pets, they have typically undergone basic obedience training with the owner (connor and miller 2000; schoenfeld-tacher, et al. 2017) and then, in order to be certified by a recognised organisation (e.g. pets as therapy, therapy dogs international, delta society), undergo a temperament assessment which assesses that the dog is sociable and friendly, calm and gentle when being stroked or handled and is not overly fearful of new and unexpected stimuli (jalongo, et al. 2004; marcus 2013). however, these dogs do not have to undergo structured training and so their behaviours cannot be as easily predicted (cavalli et al. 2017; mongillo, et al. 2015). it must be noted the differences in regulations relating to training of these different types of dogs vary across countries or jurisdictions – it is important for professionals to follow procedures and guidelines in their specific location. page 3 dogs in the cjs creative commons license 4.0 – non commercial – share alike – attribution spruin, e. & mozova, k. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science utility of different types of dogs in the criminal justice system it is indisputably clear that there are vast differences in training standards and procedures when it comes to the working role that dogs play in our society. as such, although both the level and specificity of a dog’s training should be reflected in the type of assistance they provide, there are a growing number of organisations which place dogs within environments for which they might not adequately trained. in particular, the practice of using facility dogs in supporting witnesses throughout the process of a criminal case (also known as ‘courthouse facility dog’), has been increasingly adopted across north america. this ranges from courthouse facility dogs joining witnesses for initial forensic interviews, to offering support in actual court settings (kaiser 2015). these dogs have been used across a number of settings, including vulnerable adult victims (e.g., ullman 2007) and children of different ages (e.g., parish-plass 2008), to help provide support and alleviate stress and anxiety for witnesses when providing evidence and/or testimony (crenshaw and stella 2015). the underlying aim of such service is that individuals will be providing best quality evidence, whilst at the same time their levels of discomfort will decrease. recently however, a growing number of therapy dogs have begun to take similar roles in legal settings, by comforting children throughout the legal process (e.g. bradley 2014). for example, the american humane association published the therapy animals supporting kids (task) manual which encouraged pet therapy teams to assist children in the investigation and prosecution of crimes (phillips and mcquarrie 2009). however, it seems that in this manual the authors refer to facility dogs that underwent a ‘career change’ as synonymous to therapy dogs. however, such inference does not adequately highlight the lack of training of therapy dogs and their handlers. consequently, placing therapy dogs with witnesses in legal settings, such as courtrooms and interviews, can lead to a number of liability and legal issues, as unlike facility dogs, these dogs (and handlers) might not be adequately trained within these settings and the predictability of therapy dogs, even with training, is questionable. for instance, many therapy dog organizations do not require that a dog present for evaluation to have any obedience or training classes (connor and miller 2000; schoenfeld-tacher et al. 2017). this can have repercussions both for the people the therapy dog comes in contact with, but also for the welfare of the therapy dog. zamir (2006) stated that the use of therapy dogs in any setting requires the animal to be treated as participants in a mutually beneficial relationship and that the needs of the animal must always be considered, accommodated and balanced with the needs of the client. as such, if the therapy dog shows signs of stress while a witness is testifying, the therapy dog must be removed immediately (mongillo et al. 2015), this can have clear consequences to the court proceedings and the welfare of the witness. with that, there are differences in working patterns between therapy and facility dogs. there are no clear guidelines relating to how long a therapy dog should work (mongillo et al. 2015). previous research in the area usually utilised this type of dog for short periods of time without causing distress (60 minutes maximum; e.g. glenk et al. 2014; 20 minutes palestrini et al. 2017) and the expectation is that a therapy dog should only be expected to tolerate physical intimacy with those unknown to them for minutes (glenk, 2017). the animal assisted intervention international (the only organisation aiming to facilitate global standards surrounding animal assisted intervention; couling, 2015) suggests that dogs which are not specially trained for a specific purpose (as facility dogs are) should not work more than one hour at any one time, followed by at least a half hour break. generally, there are no set guidelines as to how long a therapy dog can work for; however, it they have to be removed from a situation upon showing any signs of distress and often need to take breaks frequently (haubenhofer and kirchengast 2007; marcus 2011). this creates major issues as it is unlikely a therapy dog will be able to support an individual throughout the whole interview/testimony and may impact upon the legal process. it is impossible to provide an accurate account of how long a police interview or a court testimony lasts, as this varies depending on age or special needs of an individual, offence in question, and more (e.g. memon, et al. 2010; wilson and powell 2012). whilst the interview itself might only last for a short time, there might be frequent breaks, a waiting period, and an introduction period of page 4 www pet behaviour science org creative commons license 4.0 – non commercial – share alike – attribution 2018 | vol.5 | 1 12 the dog to the individual. the welfare of facility dogs is equally as important; however, they have been professionally trained work full working days and provide quiet companionship to vulnerable individuals in the legal setting (crenshaw and stella 2015). through their training, they enjoy being petted and enjoy resting quietly (crenshaw and stella 2015). these dogs have further been specially bred and chosen because of their calm demeanour and ability to work in high stress environments (timmins and fine 2006; weiss and greenberg 1997; wilsson and sundgren 1997), thereby decreasing the risk of creating legal issues. further to the wellbeing of the dog, only facility dogs are appropriately socialised and tested (baun et al. 2006; sachs-ericsson and merbitz, 2002). organisations responsible for certifying therapy dogs do not require contact with children during their evaluation process, rather, the evaluation focuses on and measures different aspects of the dog’s temperament (e.g., stability, shyness, aggressiveness, friendliness), and does not test dogs for safe physical contact with children; this puts children at risk for dog bites, becoming uncomfortable, and the dog becoming distressed (turner, 2006). another concern is that pet therapy organisations require that the dog’s handler always be attached to the dog by a leash (lafrance et al. 2007). this would mean that the handler must be in attendance for all private or legally sensitive matters with witnesses (e.g., forensic interviews, medical exams, therapy sessions, defence interviews, testifying). this would potentially impact on the quality of evidence gained, which is against the legal and moral objectives of acquiring evidence (e.g. cooke and davies, 2001; gordon and fleischer 2010; phillips 2015 powell, et al. 2010; sandoval 2010). further, it is questionable what the presence of the handler means in regard to the handler potentially becoming a witness to the case or being required to testify about the interview they were a part of. another issue, relevant to the use of therapy dogs in the criminal justice system, is the training of the dog handler. there is no standardised vetting or screening of handlers, rather, handlers simply need to be of ‘good character’ (not further defined) and are only trained if they choose to – often through basic obedience training they underwent with their dog (tedeschi et al. 2010). there is no screening or criminal background check carried out for handlers, which can have detrimental ramifications when working in the legal system – to the authors’ knowledge, this is something not considered in literature. it then becomes the responsibility of a specific agency to undergo such screening which can further complicate the process. in contrast, each courthouse facility dog also has a professionally trained handler working in the legal field (e.g., victim advocate, forensic interviewer, detectives, lawyer, etc), who are also carefully vetted and selected to receive one of their highly trained canines (marquad 2017). similarly to handling therapy dogs, regular re-certification of the dog/handler team is necessary. table 1. training standards: therapy vs facility dogs therapy dog type of training likely behaviour recommendation no standard training non predictable undergo appropriate training increase their socialization (individuals of different backgrounds, or ages) facility dog type of training likely behaviour recommendation standard training (> 2 years) early socialisation purposeful breeding highly predictable facility dogs can be utilised within the criminal justice system on top of issues surrounding training, certification, or handlers, there are possible challenges to consider simply due to a dog being present. one possible issue with having a dog present during interview or testimony is that the dog is used as a ‘reward’ and as such can invoke a false confession (lassiter and page 5 dogs in the cjs creative commons license 4.0 – non commercial – share alike – attribution spruin, e. & mozova, k. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science meissner 2010). whilst all interviewers are aware that the dog should not be used as such, because of the temperament and training of facility dogs, these will simply lay down by the individual and so no prompting is necessary for the dog to engage with an individual. as such, the dog serves more like a ‘comfort item’ (normally an object used to provide psychological comfort), or a ‘communication aid’ (normally an object or service used to aid individuals communicate better) special measures commonly accepted in many legal systems (e.g. cross and whitcomb 2017; hamlyn, et al. 2004). however, therapy dogs are not necessarily trained to this extent and might not engage with an individual as needed (e.g. resting beside an individual for a prolonged time period), without compromising their welfare (glenk 2017). alternatively, the authors put forward the argument that the handler might misunderstand or overstep their role and say something inappropriate, potentially portraying the dog as a reward. this leads to another issue for consideration: disruption. as stated before, criminal justice systems operate on the premise of seeking the truth and so aim to achieve best available evidence (gordon and fleischer 2010; powell et al. 2010; sandoval 2010). a therapy dog which is not trained to simply laydown and might become bored, distressed, or walk around, will likely affect the quality of evidence (disturb or distress the individual), or might potentially cause an interview/testimony to be stopped (mongillo et al. 2015; zamir 2006). as stated before, therapy dogs are not socialised and trained to the same standards as facility dogs and so there is a higher likelihood that a therapy dog will become distressed and so removed (baun et al. 2006; cavalli et al. 2017; turner 2006). lastly, there is the argument that simply having a dog present during a police interview (which might be shown as evidence in front of a jury) or during trial will automatically evoke sympathy in the jury (grimm 2013). whilst studies have shown that jurors do hold opinions about those testifying (e.g. cohen 2013), a mock juror study showed that an appropriately trained facility dog (when compared to a teddy bear or no intervention) had no effect on the jurors’ perceptions of witness credibility (burd, 2013). further exploration is needed into how the presence of a facility dog, or a therapy dog, might impact on jury/judge perceptions. however, based on available literature, the authors suggest that as facility dogs are allowed to be quietly placed into a witness box without a handler, they would not be perceived as a disruption, as compared to, therapy dogs, as they might exude more personality, move around, and similar. table 2. handler training: therapy vs facility dogs therapy dog training / selection of the handler dilemma recommendation or comments no training no standardised vetting or screening possibility of not acting appropriately in a legal situation have unsuitable background history appropriate vetted and screened specific training to such institution facility dog training / selection of the handler dilemma recommendation or comments trained vetted and screened legal professional trained handlers or trained legal professionals understand the criminal justice system environment page 6 www pet behaviour science org creative commons license 4.0 – non commercial – share alike – attribution 2018 | vol.5 | 1 12 conclusions and recommendations overall, whilst there is a very valid argument to introducing dogs, generally, into the criminal justice system processes, the differences between therapy and facility dogs need to be appreciated and so they need to be used in specific settings. following the literature discussed herein, we suggest a number of considerations for good practice when using these two different categories of dogs (tables 1, 2, 3 & 4). therapy dog guidelines no guidelines they likely work short hours with frequent breaks not trained to be utilised in an emotionally charged environment dilemma handler attached to the dog via a leash dog removed if showing signs of distress likely not be able to support an individual during the whole of a police interview/court testimony recommendation / comments therapy dogs should not be expected to fulfil a position of a working dog. their presence can be very useful in specific settings only – where they are not expected to fulfill a certain role (e.g. remain next to one person for a prolonged time period). table 3. therapy dog guidelines and recommendations facility dog guidelines trained to lay still trained to not be effected by stressful environments dilemma dogs can accompany an individual without the handler having to hold the dog’s leash recommendation / comments these dogs are able to support individuals for prolonged periods of time. therefore, they are suitable for all setting within the criminal justice system process. table 4. facility dog guidelines and recommendations we advise that therapy dogs could be introduced into some aspects of the legal process but only following a thorough selection process; they should never be allowed to enter a police interview, or a courtroom. facility dogs, however, can be utilised throughout the whole of the process, including being present during forensic/police interviewing, medical examinations, or whilst giving testimony. whilst best practice guidelines have been developed for these types of dogs separately, these guidelines have little empirical evidence behind them which calls for a need to establish an evidence base. we believe that criminal justice system users can benefit from utilising specially trained dogs in supporting them and the little available evidence is supportive of this. however, introducing such service needs to be well thought out and planned in order to truly make the criminal justice journey better for its users. page 7 dogs in the cjs creative commons license 4.0 – non commercial – share alike – attribution spruin, e. & mozova, k. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science references barker, s.b., barker, r.t., mccain, n.l. and schubert, c.m. 2016. a randomized cross-over exploratory study of the effect of visiting therapy dogs on college student stress before final exams. a multidisciplinary journal of the interactions of people and animals 29(1): 35-46. barker, s.b., knisely, j.s., schubert, c.m., green, j.d. and ameringer, s. 2015. the effect of an animal-assisted intervention on anxiety and pain in hospitalized children. a multidisciplinary journal of the interactions of people and animals, 28 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vol.5 | 1 12 recent developments in canine cognitive dysfunction syndrome. alejandro seisdedos benzal and alba galán rodríguez1 pet behaviour science | 2016, vol. 1, 47 59 alejandro seisdedos benzal and alba galán rodríguez 1. department of medicine and animal surgery. university of cordoba paper review email: ale_6d2bz@hotmail.com spain keywords: abnormal behaviour; alzheimer; cognitive disorder; cognitive dysfunction syndrome; dog highlights although canine cognitive dysfunction syndrome is an underdiagnosed disease, research in this field is growing and clinical advances have been made, and as a result, this syndrome is becoming more common. ccd causes a decline in quality of life for both the dog and the owner. it is necessary to improve the overall knowledge about this condition in order to make an early diagnosis to slow its progression. page 47creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 47 59 abstract canine cognitive dysfunction syndrome (ccd) is a neurodegenerative disease affecting aging dogs. ccd is an underdiagnosed disease that involves at least 14% of geriatric dogs, but apparently less than 2% of diseased dogs are diagnosed. there are several physiopathological similarities between alzheimer disease (ad) and ccd, developing amyloid-β deposits in brain parenchyma and blood vessels, brain atrophy and neuronal loss. the clinical signs lead to behavioural changes. they are unspecific and could appear as soon as seven years of age, but are more relevant in senior dogs. the abnormal behaviour could be classified following the acronym disha: disorientation in the immediate environment; altered interactions with humans and other animals; sleep-wake cycle disturbances; house-soiling; and changes in activity levels. there is no specific diagnostic test or biomarker to demonstrate the presence of ccd; therefore, it is often assessed by ruling out other diseases that may cause similar behavioural changes. veterinarians have to be able to make an accurate account of veterinary history asking for abnormal behaviour that could be misreported by the owners. ccd is a neurodegenerative disorder that cannot be cured. it is possible to delay the progression of the clinical signs and improve the quality of life of patients, but like in ad, the progression of the illness will depend on the individual. there are three treatment pathways, which could be used in combination: drug therapy to improve cognition and reduce anxiety, antioxidant diet and nutraceutical supplements to reduce the progression of the illness, and finally, environmental enrichment to maintain brain activity. the aim of this review article is to contribute to the knowledge of the illness, presenting recent advances in the pathophysiology, diagnosis and treatment of the disease. http://www.petbehaviourscience.org/ this review article presents the recent advances in diagnosis and treatment of the illness and contributes to our understanding of the syndrome. 1. introduction aging is a natural process that affects every cell of the body including neurons. as animals age, different illnesses that could compromise their quality of life (qol) appear. both human and veterinary medicine have increased the longevity of people and pets, respectively. therefore, the development of neurodegenerative diseases is becoming more frequent (landsberg et al. 2012). the relationship between clinical signs and physiopathology in alzheimer disease (ad) and canine cognitive dysfunction syndrome (ccd) makes the dog an excellent animal model for the understanding of ad (cummings et al. 1996; fast et al. 2013). canine ccd is an age-related neurodegenerative illness that generally affects dogs over seven years of age and is characterized by gradual cognitive decline and increasing brain disorders (landsberg 2005; landsberg et al. 2011, laflamme 2012; nagasawa et al. 2014). the age of onset of the clinical signs depends on the breed according to their life span differences. despite the fact that the prevalence did not differ between breeds (salvin et al. 2006), smaller breeds have a longer life span than the larger ones, thus it is possible to recognize more clinical signs of ccd in small breeds (vite and head 2014; schmidt et al. 2015). ccd prevalence is reported to range from 14% (salvin et al. 2006) to 22% (azkona et al. 2009). in a study of 497 dogs from eight to 19 years, salvin and colleagues (2006) recruited owners to an internet-based survey. this 84-item questionnaire was used to estimate the incidence of ccd. the survey included questions about activity levels, behaviour, phobias, eating and drinking habits, management and health. they found an overall prevalence of 14.2%, but it could be up to 41% in dogs older than 14 years. consequently, salvin et al. (2006) suggested that the prevalence seems to be higher in older dogs. azkona et al. (2009) used 325 geriatric dogs to describe the prevalence of the disease. they used phone interviews to gather information about four behavioural categories related to cognitive impairment: sleep-wake cycles, social interaction, learning and house training and signs of disorientation. regardless of breed, the prevalence seems to be higher in females than in males. prevalence is also reportedly higher in neutered males, than entire males (landsberg et al. 2011). there are numerous nonspecific clinical signs of ccd, like disturbance of the sleep/wake cycle, changes in social interactions with people, urinary and faecal incontinence, disorientation in well-known places, hearing loss, and excessive vocalization among others (dewey 2008, p. 123; nagasawa et al. 2014). because of these nonspecific clinical signs and the fact that the owners usually think that the behavioural changes of their pets are due to the typical changes caused by aging, it is often difficult to make an accurate and early diagnosis of ccd. recent studies have focused on the development of diagnostic tools based on food searching or problem solving tasks. there are also recent advances in the knowledge of the physiopathology of the aging brain, and about new treatments with drugs, diets and environmental enrichment (landsberg et al. 2011; laflamme 2012; landsberg et al. 2012; gonzález-martínez et al. 2013; vite and head 2014). it is documented that cognitive impairment in older dogs can be a challenging problem, both for the animal and for the owners. the high percentage of geriatric cognitive impaired dogs and the reduction in dog’s qol suggest a better knowledge of ccd is necessary (landsberg et al. 2011). the aim of this review is to compile the recent advances in physiopathology, diagnosis and treatment options for ccd. 2. physiopathology there are several histopathologic similarities between human brains affected by ad and dog brains suffering from ccd. in both diseases, the aged brain develops an abnormal β-amyloid (aβ) deposit in brain parenchyma and blood vessels, neuronal loss and hyperphosphorylated tau protein (htp) (schmidt et al. 2015). aβ is a protein produced by the degradation of amyloid precursor protein (app) (vite and head 2014). in the aging dog brain the deposits present as diffuse plaques in cortical regions, and tend to appear from eight years of age (landsberg et al. 2012; fast et al. page 48 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 47 59 2013; schmidt et al. 2015). the prefrontal cortex is the first area affected, followed by the temporal cortex, the hippocampus and the occipital cortex. regardless of position, the amount and extent of aβ deposits are correlated with the degree of cognitive impairment and leads to oxidative damage (fast et al. 2013; vite and head 2014; romanucci and della salda 2015; schmidt et al. 2015). in addition, a local inflammatory reaction has been found around aβ deposits in both humans and dogs (schmidt et al. 2015). aβ deposits are also found in brain blood vessels in ad and ccd. cerebrovascular amyloid angiopathy (caa) is related to cognitive impairment in both humans and dogs (romanucci and della salda 2015). the occipital cortex is the main affected area and these deposits could disrupt the blood-brain-barrier and cause microhemorrhages (vite and head 2014). the brain consumes 20% of the total body´s oxygen and is very susceptible to oxidative damage due to the high percentage of polyunsaturated fatty acids and the lower levels of endogenous antioxidant activity (head 2008). the metabolic process of the cells causes the release of reactive oxygen species (ros) that lead to oxidative damage to proteins, lipids, dna and rna (head 2008). the production of ros depends on the mitochondria (romanucci and della salda 2015); therefore, a mitochondrial dysfunction could cause the overproduction of ros. this, together with a lesser activity of antioxidants enzymes in the aged canine or in the neurodegenerative disease could lead to oxidative damage, aβ deposition and cognitive dysfunction (dowling and head 2012; davis and head 2014; vite and head 2014; romanucci and della salda 2015). iwata and colleagues (2005) described the aβ catabolism and found that an imbalance between production and clearance of aβ could provoke the accumulation of aβ in the brain. aβ could be eliminated by two mechanisms: active transport through the blood brain barrier and enzymatic degradation by peptidases like neprilysin (npl) (canudas et al. 2014). in the brain areas in which the aβ deposits are more frequent, npl is expressed in lower levels than areas with lesser amounts of aβ accumulation (reilly 2001). canudas et al. (2014) found that the expression of npl was five times higher in the cognitive unimpaired dogs than in the cognitive impaired dogs. this suggests that npl could protect against ccd. in a recent study, schmidt and colleagues (2015) provided the first description and quantification of pyroglutamyl aβ (pe3aβ) in the canine brain. the pe3aβ is a peptide derived from aβ peptides that suffers from n-terminal truncation and subsequent cyclization of n-terminal glutamate (hartlagerübsamen et al. 2011). this peptide encourages the accumulation of aβ facilitating the onset of ccd. the ventral hippocampus and entorhinal cortex of aged dogs’ brain seem to be more affected by pe3aβ deposits; the location of these deposits correlates with clinical signs of ccd, such as disorientation and memory loss (schmidt et al. 2015). in human medicine there is a documented relationship between htp and cytotoxicity of aβ deposits (schmidt et al. 2015). in a study by schmidt et al. (2015), only one out of 24 dogs displayed formation of neuro fibrillary tangles of htp. therefore, research reinforces that the aβ deposits and the htp are two independent processes in the aged canine brain (head 2002; schmidt et al. 2015). other physical pathological changes in ccd are different processes related to brain atrophy, such as: thinning of the cerebral cortex and subcortical white matter; widened sulci; marked ventriculomegaly; meningeal calcification; demyelination; increased lipofuscin; neuroaxonal degeneration; and apoptotic bodies (laflamme 2012; landsberg et al. 2012; vite and head 2014). brain atrophy may be due to neuronal loss. in the brain of dogs, new neurons are produced continually from progenitor cells placed in specific brain regions such as the dentate gyrus of the hippocampus. in a previous study, siwak-tapp and colleagues (2007) found a 90% to 96% decline in neurogenesis in the hippocampus of aged beagles. this suggests that the neuronal loss in the brain atrophy process could be related to cognitive performance in aged dogs. according to pan and colleagues (2012), reduction in neurogenesis could be caused by the lesser extent of progenitor cell page 49 canine cognitive dysfunction syndrome creative common license 4.0 – non commercial – share alike – attribution seisdedos benzal, a. and galán rodríguez, a. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science proliferation and the shorter survival of the neurons. alterations in neurochemical systems such as the cholinergic, glutaminergic, dopaminergic and gabaergic neurotransmitter systems, with reduced neuronal and synaptic function have also been observed (landsberg et al. 2012). all these brain changes could contribute to the primary clinical signs of ccd like memory loss, alteration in spatial orientation, and alteration in rem sleep (landsberg et al. 2012). 3. clinical signs when dogs are aged seven years old or older, we consider that they are senior canines at risk of developing an age-related health problem (laflamme 2012). although clinical signs of ccd may appear at seven years old, these become more relevant as dogs age. nevertheless, owners often overlook ccd signs, dismissing them as normal aging. delay in the recognition of cognitive impairment could be due to the fact that the owners sometimes do not realize subtle signs of behavioural changes or associate the changes with the advanced age of their pets. it is very important to consider the complete veterinary history and to question dog owners/handlers about behavioural changes that could be indicative of cognitive impairment when diagnosing the condition (landsberg et al. 2012). vite and head (2014) describe a case of ccd in an 11 year old female border collie used on a farm as a livestock herding dog. according to her owner, when the dog reached 11 years, she was almost deaf, had begun to show some confusion with commands and had lost interest in pursuing sheep. over the next two years the hearing loss progressed and her behaviour was perceived by the owner as stubbornness. she started to stalk the other resident dog and began pacing, panting, and showing signs of anxiety when her owner was not at home. she was no longer making eye contact with people and sometimes she lost the route that she had previously followed daily. she was euthanized at 14 years old and an extensive diffuse aβ deposit was found in the dentate gyrus and prefrontal cortex. all the physiopathological changes in ccd cause several clinical signs that are classified following the acronym disha (table 1): disorientation in the immediate environment, altered interactions with humans and other animals, sleep-wake cycle disturbances, house-soiling, and changes in activity levels (manteca 2011; gonzález-martínez et al. 2013). in addition, it is important to consider that there is individual variability between aged dogs in terms of clinical signs for the same pathology (nagasawa et al. 2014). despite this, in a study by mariotti et al. (2009), aggression was the most reported clinical sign (53%), followed by fear and anxiety reported in a much lower page 50 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 47 59 table 1. clinical signs of ccd that can be classified following the acronym disha proportion of subjects. to summarise, in cases of suspected ccd, it is essential that the veterinarian assesses for clinical signs related to disha. dogs with ccd may also express signs of anxiety; however, these may also appear with pain or relate to other systemic illnesses. these signs could be new behavioural patterns like eliminating in unusual places at home, cessation of jumping behaviour when greeting people, avoidance of the owner or home residents, unusual growling or aggressiveness, excessive panting, yawning, nose licking, or momentary immobilization. social alterations related to anxiety, such as decreased play-time with people and other pets, decreased interest in toys, altered quality of sleep, and hiding behaviour may also be relevant. when signs of anxiety appear at night it may pose a significant problem due to sleep disruption of the home residents, thus the relationship between owner and pet may become negatively affected (landsberg et al. 2011). nearly all of the clinical signs in the disha suite could be confused with different diseases. in this way, veterinarians have to be cautious when diagnosing ccd without ruling out other pathologies. disorientation in the immediate environment may be caused by hearing loss as a result of aging or chronic otitis, visual loss because of the development of cataracts, glaucoma or retinal detachment among others (bernardini 2010, p. 30, 36). changes in interactions with humans or other animals could be due to some painful illnesses. it is frequent that senior dogs developing arthritis and/or arthrosis are reluctance to move and play with other dogs, and may even become aggressive in an attempt to avoid contact (nelson and couto 2009, p. 1121). epileptic dogs can show an abnormal behaviour during the preand postseizure period (de risio and platt, 2014, p.39). the allodynia or hypersensitivity to skin pressure due to neuropathic pain could also deteriorate the interaction with humans. in this case, when owners caress the affected region the dog may even yell or moan due to pain (grubb 2010). sleep-wake cycle disturbances and house-soiling are also related to systemic illnesses, like renal or hepatic failure. the development of acute or chronic kidney disease is common in geriatric dogs. these diseases may provoke changes to urination patterns, pain and a reduction on the activity of the dog (nelson and couto, p. 491, 617). there are several disorders that may change the normal activity of the dog. metabolic disorders like hypothyroidism and hypoadrenocorticism could cause, among other symptoms, lethargy, reluctance to move and tendency to gain weight (ettinger and feldman 2010). cardiovascular disorders like dilated or hypertrophic cardiomyopathy, heart blockage or valve heart disease, among others, could cause a reluctance to move or quick fatigue during exercise (ettinger and feldman 2010,). to the authors’ knowledge, disha constitutes the best model to explain the clinical signs of ccd since it puts together the main behavioural changes of a cognitively impaired dog. anxiety might be a new letter to add to the acronym but we consider that all the behavioural changes of disha could cause some discomfort and anxiety to the dog. 4. diagnosis salvin and colleagues (2011) conducted an epidemiological study surveying 497 dogs ranging from eight to 19 years. the overall prevalence of ccd was calculated based on owner reports to be 14.2%; however, only 1.9% had a presumptive diagnosis of ccd from a veterinarian. underdiagnosis may be frequent due to the lack of a specific diagnostic tool and the unspecific symptoms. as commented previously, some owners are not able to report subtle behavioural changes in their pets or may have dismissed them as typical aging processes. therefore, veterinarians must be active in asking about any changes or abnormalities to the dog´s behaviour, asking specifically for disorientation in daily life, longer sleeping time along the day, loss of house training and house-soiling. it is also important to know the normal behaviour of the dog during its life to assess any potential abnormal changes in behaviour due to an underlying disease. there are several types of chronic illnesses that may provoke changes in canine behaviour. orthopaedic disease could cause pain, cardiovascular impairment may lead to reluctance to exercise and metabolic disorders could reduce the normal activity of the animal or increase aggressiveness. to ensure accuracy, page 51 canine cognitive dysfunction syndrome creative common license 4.0 – non commercial – share alike – attribution seisdedos benzal, a. and galán rodríguez, a. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science the diagnosis of ccd should be made following exclusion of all these diseases (landsberg et al. 2011). therefore, it is necessary to make an exhaustive clinical exam (physical, neurological, ophthalmologic, orthopaedic and ethological examination) to assess behavioural changes related to any physical illness (landsberg et al. 2012). intracranial pathologies related to behavioural changes, such as brain tumours, are more frequent in aged dogs. endocrine disorders could cause irritability, lethargy and aggressiveness. metabolic disorders like hepatic or renal failure lead to an altered mentation and house-soiling. orthopaedic, gastrointestinal, urogenital and dermatological diseases likewise neuropathic pain could cause severe pain conditions that may provoke behavioural changes. cardiovascular diseases are related to restless (landsberg et al. 2011). in addition, it is common to find two or more of these diseases in geriatric patients. knowledge of existing and/or previous medications and treatments is also important, and possible cognitive impairment associated with these drugs should be ruled out. steroids have been shown to be associated with changes to eating, drinking and urination as well as nervousness and aggression. in addition, pain medication can sometimes cause sedation (landsberg et al. 2012). when all these pathologies have been ruled out it is important to carry out an owner-based observational questionnaire to assess the cognitive function of the patient (gonzález-martínez et al. 2013). nielson and colleagues (2001), carried out a study with 180 dogs from 11 to 16 years old, using a questionnaire including items related to four categories of cognitive impairment (disorientation, altered social interactions, disturbed sleep-wake cycle and house training). they assessed the severity of the cognitive impairment depending on the number of categories affected and the number of new behavioural changes found in each category. to the authors´ knowledge, it could be essential to include items related to all categories of disha in any ownerbased observational questionnaire. because there is individual variability in the presentation of clinical signs and, due to the fact that disha includes all the behavioural changes known to be found in a dog with ccd, a questionnaire based on disha could be an appropriate diagnostic tool. gonzález-martínez et al. (2013), searched for a more objective diagnostic procedure than these owner-based observational questionnaires. they assessed the effectiveness of two simple tasks (a food searching task [fs] and a problem solving task [ps]) administered in a clinical setting in 87 dogs. they found differences in the two tasks between the cognitively impaired group and the young-middle aged group. the younger dogs comprising the second group were able to find and obtain the food more quickly than their aged counterparts in the cognitively impaired group. in addition, some dogs in the latter group did not even attempt to search for the food. these assessment tasks could help in the accurate diagnosis of ccd. 5. treatment it is important for veterinarians to advise owners of dogs diagnosed with ccd that it is a neurodegenerative disorder that cannot be cured, although it is possible to delay the progression of the clinical signs and improve the qol of the dog. however, as in ad, the advancement of the illness is dependent on the individual (landsberg et al. 2011). current treatments tend to combine specific diets, environmental enrichment and drugs, obtaining better success and improving pets´ qol. the sooner treatment begins, the better the response, therefore it is important to make an early diagnosis (landsberg et al. 2012). nutritional treatment in human medicine, there are several studies about the best diet option for the treatment of ad. the combination of various antioxidants seems to be more efficacious as part of the diet than as a single nutraceutical supplementation (dowling and head 2012). in veterinary medicine, studies shown that an antioxidant diet, in addition to some dietary supplements, is the best nutritional therapy (laflamme 2012; landsberg et al. 2012)(table2). page 52 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 47 59 initially, the nutrition of the pet’s diet must be evaluated because of the changes in the nutritional requirements associated with aging. maintenance energy requirements decrease about 25% in dogs greater than seven years of age and brain glucose metabolism is also reduced. thus it is mandatory to adapt the current diet to their current nutritional requirements (laflamme 2012). dowling and head (2012) used an antioxidant diet formulated with vitamins a, c and e, l-carnitine, lipoic acid, zinc, selenium and dehydrated fruits and vegetables (spinach flakes, tomatoes, grape pomace, carrots and citrus pulp in a 1% proportion each). they proved that reduced oxidative stress leads to cognitive benefits. they used 48 senior beagles divided in four groups: two control diet groups, one with and the other without behavioural enrichment (groups 1 and 2), and other two groups of antioxidant diets with and without behavioural enrichment respectively (groups 3 and 4). there were significant improvements over a two week period related to spatial attention, visual discrimination and learning ability in the groups fed with antioxidant diet, especially in the group with environmental enrichment. in addition, reduced oxidative damage and increased endogenous antioxidant activity were found in groups 3 and 4 in the study. glucose is the main energy source of neurons. however, glucose metabolism is reduced with aging and other energy sources are needed to maintain neurons´ metabolism (pan 2011; laflamme 2012). fatty acids derived from mct could provide up to 20% of brain energy request. for instance, lactate and ketones cross the blood-brain barrier and serve as energy source (laflamme 2012; landsberg et al. 2012). pan and colleagues (2010) found significant improvement in cognitive function in dogs fed with medium chain triglycerides. it has been proven that diets supplemented with this type of lipid also reduce levels of app and aβ deposits and improve mitochondrial function (taha et al. 2009). nowadays, we can find numerous commercial diets containing medium chain triglycerides and antioxidants such as hill's pet nutrition b/d and purina one vibrant maturity 7+ senior. due to the possibility of two or more concomitant diseases in senior dogs, often it is not possible to make changes in the diet. in this case the use of single nutraceuticals can be an alternative. phosphatidylserine is a phospholipid that improves social interactions, house soiling and orientation in dogs (heath et al. 2007). in a study of aged beagles fed with phosphatidylserine, gingko biloba, vitamin e and pyridoxine an improvement in their short term memory was observed (dowling and head 2012). sadenosyl-l-methionine increases endogenous production of antioxidant glutathione and improves neural conduction enhancing executive function (landsberg et al. 2012). apoaequorin is a protein that page 53 canine cognitive dysfunction syndrome creative common license 4.0 – non commercial – share alike – attribution seisdedos benzal, a. and galán rodríguez, a. table 2. diet and dietary supplementation used in ccd (landsberg et al. 2012). https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science seems to have neuroprotection hallmarks in aging dogs. this protein improves learning and attention in laboratory trials (milgram et al. 2011). docosahexaenoic acid (dha) and eicosapentaenoic acid (epa) are two polyunsaturated omega-3 fatty acid present in fish oil that protects the normal neural functions. it is seen that these fatty acids decrease in ad but there are no data about the beneficial properties in ccd (laflamme 2012). galán and colleagues (2014) used a nutritional supplement (aktivait®, vetplus) that includes n-acetil cysteine, a-lipoic acid, vitamins c and e, l-carnitine, co-enzyme q10, and polyunsaturated fatty acids (pufa, phosphatidylserine, dha and epa) in order to evaluate changes in cerebrospinal fluid after a 50-dayperiod of treatment. they found increased levels of glucose and sodium and lower levels of lactate and lactate/pyruvate ratio, suggesting an improvement in brain energy metabolism after dietary supplementation. there are several studies about the beneficial effects of acetylcarnitine and lipoic acid in ad and ccd. both of them enhance mitochondrial function reducing reactive oxygen species and also reducing oxidative damage. nevertheless, when these two products are used in combination with antioxidant substances, the beneficial effects are not seen (dowling and head 2012). alphacasozepine is a biopeptide from milk that is proved to reduce anxiety in rodents and it has been used as a nutraceutical compound to treat anxiety and behavioural disorders like aggression in dogs (manteca 2011). in summary, an antioxidant diet combined with nutraceuticals (mct, phosphatidylserine, apoaequorin, dha and epa) improves cognitive impairment in dogs. this treatment enhances visuospatial function, learning ability and the ability to focus the attention. environmental enrichment it has been demonstrated that behavioural enrichment is protective against neuron loss in the hippocampus of dogs, improving all disha parameters (dowling and head 2012). the enrichment has to include physical exercise, social interactions and cognitive training (romanucci and della salda 2015). mental activity is an important component to maintain a good qol. effective strategies to increase the attention of cognitively impaired dogs include the use food/treat puzzle toys, walks in new environments and to play games such as fetch and chase. in using environmental enrichment, it is also important to improve the comfort of the pet. new odour, tactile and sound cues could help the dog to orientate in the environment. if there is an increase in the urine frequency, it may be necessary to increase the number of walks or to put a toilet area inside home (landsberg et al. 2012). when there are problems of night time waking and anxiety, it is important to improve dog´s sleeping patterns. owners should pet the dog when it is resting in its sleeping area, turn off the tv, reduce noises and intensity of lights during the night, and place the dog´s bed in its preferred resting area. a useful routine to enhance the sleeping time is to schedule a physical activity such as game playing, or a long walk to tire the dog, before to the dog’s rest time (landsberg et al. 2011). drug therapy the aim of the drug therapy is to restore the natural brain function and to slow the progression of the illness. due to the fact that an inflammatory reaction around aβ deposits exists, it has been proposed the use of anti-inflammatory drugs like carprofen (4,5mg/kg/24h) (dewey 2008, p. 126). there are several medications for the treatment of ccd (see tables 3 and 4 for the main and complimentary drugs used in the treatment of ccd respectively). traditionally, the three drugs selegiline, nicergoline and propentofylline, have been used for the treatment of ccd. they confer beneficial effects in activity levels, improving catecholamines´ levels in the cortex and also the blood flow to the brain. propentofylline also increases spatial attention and it is effective treating dullness, lethargy and depression; whereas selegiline also enhances social interactions and sleep-wake cycles (landsberg et al. 2012). however it is not possible to enhance all the disha categories by using these drugs alone (landsberg et al. 2012). there are other drugs such as adrafanil and modafinil, two noradrenergic-enhancing drugs, that improve locomotion and learning in dogs (dewey 2008, p. 126). inagawa and colleagues (2005) found that γpage 54 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 47 59 aminobutyric acid (gaba) improves qol of cognitive impaired dogs. different behavioural changes like whines, barks during the night and when the dog is home alone, wandering, and sleeping patterns were all effectively/moderately alleviated with the use of a test preparation containing gaba. a gaba-ergic drug that could be used for the treatment of ccd is gabapentin (dewey 2008, p. 126; landsberg et al. 2012). a recent study by nagasawa and colleagues (2014) showed that n-acetyl-d-mannosamine (mannac) alleviates clinical signs of ccd. four out of the five dogs treated with mannac showed a permanent improvement in sleep quality. to the authors’ knowledge, selegiline is the most useful drug in treating ccd (jaggy 2010, p.485), however it is necessary to combine this with complimentary therapy in order to enhance all the behavioural changes associated to the physiopathology of the syndrome. benzodiazepines are useful to treat sleep-wake cycle disturbances and anxiety because of their sedative and short-acting anxiolytic effects (landsberg et al. 2011). these substances are often used to treat fears and phobias and can also help the dog to sleep better. phenobarbital, diphenhydramine and trazodone may also be used for sleep support due to their sedative effect. it is recommended these drugs be given 30 minutes before sleeping (landsberg et al. 2011). other clinical signs related to anxiety, such as noise and night phobias and separation anxiety, could be treated with buspirone, fluoxetine or tricyclic antidepressants. nevertheless, the anticholinergic effects need to be taken into consideration (jaggy 2010, p. 487, landsberg et al. 2011). there is some controversy surrounding the use of statins for the treatment of ccd. atorvastatin is a selective inhibitor of hmg-coa reductase that acts lowering cholesterol levels and reducing aβ deposits. this drug has been used in human medicine reducing the risk of developing ad. however, it has been shown that atorvastatin reduces levels of coenzyme q10 (coq10), a mitochondrial electron transporter and a page 55 canine cognitive dysfunction syndrome creative common license 4.0 – non commercial – share alike – attribution seisdedos benzal, a. and galán rodríguez, a. table 3 main drugs and doses for ccd treatment table 3. maind drugs and doses for ccd treatment https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science powerful cellular antioxidant. low levels of coq10 in the brain could be related to cognitive impairment (martin et al. 2011). as seen in martin and colleagues (2011) study, serum coq10 was significantly lower in atorvastatin-treated dogs. therefore, more studies are needed to clarify the relationship between atorvastatin and coq10 and to determine the effectiveness of a combined treatment with atorvastatin and coq10 oral supplementation. in a recent study, bosch and colleagues (2015) developed an anti-aβ immunotherapy that modified the equilibrium between soluble and insoluble aβ in aged canines. in the immunized group, the amyloid plaques were smaller and less compact than those of the unimmunized group. among natural products that could help reduce anxiety and improve qol for canine with ccd, the valerian root, honokiol, berberine extracts, α-casein, suntheanine, lavender essential oils, and a pheromone collar have been shown to be useful (dewey 2008, p. 126; landsberg et al. 2011). to conclude, it is important to emphasise the need to use a combined therapy to treat ccd. according to disha, d and h parameters are specially improved with the nutritional treatment using an antioxidant or mct enriched diet (i and a parameters could be also improved with an alpha-casozepine and s-adenosyl-lmethionine supplementation, respectively). s and a parameters and anxiety could be enhanced by using different drugs. finally, environmental enrichment helps in all the disha categories but especially in i parameter. the stimulation of mental activity along with physical exercise, cognitive training and games, improves social interactions with both humans and other animals. summary ccd is an underdiagnosed neurodegenerative disease that affects a high proportion of the aged canine population. the number of unspecific clinical signs makes it difficult to diagnose and there are no specific biomarkers of the illness. therefore, the veterinarian has to rule out other diseases and medical treatments that may cause behavioural changes associated with the acronym disha. there are numerous choices for the treatment of ccd but some of these require more research to clarify their effectiveness. however, the best treatment option we currently know of is to combine drug and nutritional therapy with an environmental enrichment program to improve the qol of both the owners and the canines. page 56 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 47 59 table 4: complimentary drugs and doses used in the treatment of ccd table 4. complimentary drugs and doses used in the treatment of ccd 6. references azkona, g., garcía-belenguer, s., chacón, g., rosado, b., león, m., and palacio, j. 2009. prevalence and risk factors of behavioural changes associated with agerelated cognitive impairment in geriatric dogs. journal of small animal practice 50: 87-91. bernardini, m. 2010. esame neurologico. in neurologia del cane e del gatto, 30-69, ed. m. bernardini. milano: poletto editore. bosch, m.n., pugliese, m., andrade, c., gimeno-bayón, j., mahy, n., and rodríguez, m.j. 2015. amyloid-β immunotherapy reduces amyloid plaques and astroglial reaction in aged domestic dogs. neurodegenerative diseases 15(1): 24-37. canudas, j., insua, d., sarasa, l., gonzález-martínez, á., suárez, m.l., santamarina, g., pesini, p., and sarasa, m. 2014. neprelysin is poorly expressed in the prefrontal cortex of aged dogs with canine cognitive dysfunction syndrome. international journal of alzheimer ´s disease 2014. cummings, b.j., head, e., 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disorders. in textbook of veterinary internal medicine, 1711-1873, ed s.j. ettinger and e.c. feldman. st. louis, missouri: saunders elsevier. ettinger, s.j. and feldman e.c. 2010. pathophysiology of heart failure. in textbook of veterinary internal medicine, 1143-1158, ed s.j. ettinger and e.c. feldman. st. louis, missouri: saunders elsevier. fast, r., rodell, a., gjedde, a., mouridsen, k., alstrup, a.k., bjarkam, c.r., west, m.j., berendt, m., and moller, a. 2013. pib fails to map amyloid deposits in cerebral cortex of aged dogs with canine cognitive dysfunction. frontiers in aging neuroscience 5: article number 99. galán, a., carletti, b.e., morgaz, j., granados, m.m., mesa, i., navarrete, r., lombardo, r., martínez, c.m., and martín-suárez, e.m. 2014. comparative study of select biochemical markers in cerebrospinal fluid of healthy dogs before and after treatment with nutraceuticals. veterinary clinical pathology 43(1): 72-77. gonzález-martínez, á., rosado, b., pesini, p., garcíabelenguer, s., palacio, j., villegas, a., suárez, m.l., santamarina, g., and sarasa, m. 2013. effect on age and severity of cognitive dysfunction on two simple tasks in pet dogs. the veterinary journal 198: 176-181. grubb, t. 2010. chronic neuropathic pain in veterinary patients. topics in companion animal medicine 25(1): 4552. hartlage-rübsamen, m., morawski, m., waniek, a., jäger, c., zeitschel, u., koch, b., cynis, h., schilling, s., schliebs, r., demuth, h.u., and robner, s. 2011. glutaminyl cyclase contributes to the formation of focal and diffuse pyroglutamate (pglu)-aβ deposits in hippocampus via distinct cellular mechanisms. acta neuropathologica 121: 709-719. page 57 canine cognitive dysfunction syndrome creative common license 4.0 – non commercial – share alike – attribution seisdedos benzal, a. and galán rodríguez, a. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science head, e. 2002. brain aging in dog: parallels with human brain aging and alzheimer’s disease. journal of veterinary pharmacology and therapeutics 2(3): 247-260. head, e. 2008. oxidative damage and cognitive dysfunction: antioxidant treatments to promote healthy brain aging. neurochemical research 34(4): 670-678. heath, s.e., barabas, s., and craze, p.g. 2007. nutritional supplementation in cases of canine cognitive dysfunction-a clinical trial. applied animal behaviour science 105: 274–83. inagawa, k., seki, s., bannai, m., takeuchi, y., mori, y., and takahashi, m. 2005. alleviate effects of γaminobutyric acid on behavioural abnormalities in aged dogs. the journal of veterinary medical science 67(10): 1063-1066. iwata, n., higuchi, m., and saido, t.c. 2005. metabolism of amyloid-β peptide and alzheimer´s disease. pharmacology and therapeutics 108: 129-148. jaggy, a. 2010. behavioural problems and abnormal behaviour. in small animal neurology, 467-489, ed. a. jaggy. germany: schlütersche. laflamme, d.p. 2012. nutritional care for aging cats and dogs. veterinary clinics of north america-small animal practice 42: 769-791. landsberg, g. 2005. therapeutics agents for the treatment of cognitive dysfunction syndrome in senior dogs. progress in neuro-psychopharmacology and biological psychiatry 29: 471-479. landsberg, g., deporter, t., and araujo, j.a. 2011. clinical signs and management of anxiety, sleeplessness and cognitive dysfunction in the senior pet. veterinary clinics of north america-small animal practice 41: 565-590. landsberg, g., hunthausen, w., and ackerman, l. 2013. appendix d drug dosages. in behaviour problems of the dog and cat, 415-422, ed. r. edwards. china: saunders elsevier. landsberg, g.m., nichol, j., and araujo, j.a. 2012. cognitive dysfunction syndrome. a disease of canine and feline brain aging. veterinary clinics of north america-small animal practice 42: 749-768. manteca, x. 2011. nutrition and behavior in senior dogs. topics in companion animal medicine 26: 33-36. mariotti, v.m., landucci, m., lippi, i., amat, m., manteca, x. and guidi, g. 2009. epidemiological study of behavioural disorders in elderly dogs [abstract]. in proceedings 7th international meeting of veterinary behaviour medicine, 241-243, ed. s. e. heath. belgium: european society of veterinary clinical ethology. martin, s.b., cenini, g., barone, e., dowling, a.l.s., mancuso, c., butterfield, d.a., murphy, m.p., and head, e. 2011. coenzyme q10 and cognition in atorvastatin treated dogs. neurosciences letters 501: 9295. milgram, n.w., landsberg, g.m. and visnesky, m. 2011. effect of apoaequroin on cognitive function in aged canines. paper presented at the 17th congress of esvce “social communications in companion animals” and 1st annual congress of ecawbm, avignon, france, november 25-26, 2011. nagasawa, m., shimozawa, a., mogi, k., and kikusui, t. 2014. n-acetyl-d-mannosamine treatment alleviates age-related decline in place learning ability in dogs. journal of veterinary medical science 76: 757-761. nelson, r.w. and couto c.g. 2009. clinical manifestations of hepatobiliary disease. in small animal internal medicine, 485-495, ed. r.w. nelson, c.g. couto. st, louis, missouri: mosby elsevier. nelson, r.w. and couto c.g. 2009. clinical manifestations of urinary disorders. in small animal internal medicine, 607-622, ed. r.w. nelson, c.g. couto. st, louis, missouri: mosby elsevier. nelson, r.w. and couto c.g. 2009. disorders of the joints. in small animal internal medicine, 1127-1141, ed. r.w. nelson, c.g. couto. st, louis, missouri: mosby elsevier. nielson, j.c., hart, b.l., cliff, k.d., and ruehl, w.w. page 58 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 47 59 2001. prevalence of behavioral changes associated with age-related cognitive impairment in dogs. journal of the american veterinary medical association 218: 1787-1791 pan, y., larson, b., araujo, j.a., lau, w., de rivera, c., santana, r., gore, a., and milgram, n.w. 2010. dietary supplementation with medium-chain tag has longlasting cognition-enhancing effects in aged dogs. british journal of nutrition 103: 1746–1754. pan, y. 2011. enhancing brain function in senior dogs: a new nutritional approach. topics in companion animal medicine 26: 10-16. pugliese, m., gangitano, c., ceccariglia, s., carrasco, j.l., del fa, a., rodríguez, m.j., michetti, f., mascort, j., and mahy, n. 2007. canine cognitive dysfunction and the cerebellum: acetylcholinesterase reduction, neuronal and glial changes. brain research 1139: 85–94. reilly, c.e. 2001. neprilysin content is reduced in alzheimer brain areas. journal of neurology 248: 159-160. romanucci, m. and della salda, l. 2015. oxydative stress and protein quality control systems in the aged canine brain as a model for human neurodegenerative disorders. oxidative medicine and cellular longevity 2015. salvin, h.e., mcgreevy, p.d., sachev, p.s., and valenzuela, m.j. 2006. under diagnosis of canine cognitive dysfunction; a cross-sectional survey of older companion dogs. veterinary journal 184: 277-281. schmidt, f., boltze, j., jäger, c., hofmann s., willems, n., seeger, j., härtig, w., and stolzing, a. 2015. detection and quantification of β-amyloid, pyroglutamil aβ, and tau in aged canines. journal of neuropathology and experimental neurology 74: 912-923. siwak-tapp, c.t., head, e., muggenburg, b.a., milgram, n.w., and cotman, c.v. 2007. neurogenesis decreases with age in the canine hippocampus and correlates with cognitive function. neurobiology of learning and memory 88(2): 249–259. taha, a.y., henderson, s.t., and burnham, w.m. 2009. dietary enrichment with medium chaintriglycerides (ac-1203) elevates polyunsaturated fatty acids in the parietal cortex of aged dogs; implications for treating age-related cognitive decline. neurochemical research 34(9): 1619–1625. vite, c.h. and head, e. 2014. aging in the canine and feline brain. veterinary clinics of north america-small animal practice 44: 1113-1129. page 59 canine cognitive dysfunction syndrome creative common license 4.0 – non commercial – share alike – attribution seisdedos benzal, a. and galán rodríguez, a. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science does cat attachment have an effect on human health? a comparison between owners and volunteers filipa alexandra benchimol da silva garcia dinis and thaís lima fernandes martins1 highlights physiological responses are reported before, during and after cat presence the effect of reported levels of attachment for both owners and volunteers is investigated pet behaviour science | 2016, vol. 1, 1 – 12 filipa alexandra benchimol da silva garcia dinis thaís lima fernandes martins 1. cornwall college newquay. centre for applied zoology paper research email: thais.martins@cornwall.ac.uk tr7 2lz united kingdom keywords: attachment, companion animals, cat, human-animal interaction (hai), human health the importance of pet ownership for health benefits, particularly long-term ownership, is highlighted questions about the possible effect of the combined effect of home and ownership in providing health benefits are raised 1. introduction there is a growing scientific interest across a range of disciplines in the relationship between pets and humans, which is often referred to as human animal interaction (hai). there is growing evidence of benefit in particular when related to physical and psychological effects (reviews in chur-hansen et al. 2010; arhant-sudhir et al. 2011; beetz et al. 2012). owning a pet is associated with reduced stress (e.g. allen et al. 1991; allen et al. 2002; beetz et al. 2011), lower salivary cortisol (e.g. beetz et al. 2011), lower heart rate and blood pressure (e.g. nagengast et al. 1997; demello 1999), and improved self-esteem (e.g. mcnicholas et al. 2005). reductions in stress seem to involve reduction in activity in the central autonomic nervous system (allen et al 1991; arhant-sudhir et al. page 1creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 1 12 abstract cat owners and volunteers from a rehoming centre were given the lexington attachment to pet scale (laps) questionnaire to assess their level of attachment to their own or rescue cats. in addition, heart rate and blood pressure were measured 10 minutes before, during, and after spending time with the cats. consistent with other studies, the results here show that spending time with a cat can reduce heart rate and both systolic and diastolic blood pressure, and that this reduction is generally more pronounced in the cats’ owners rather than in volunteers from a cat rehoming centre. for owners, levels of attachment as measured by the laps scale were positively associated with this reduction in metabolic measurements before and during pet presence; i.e. the difference (b-d) was positively correlated with the level of attachment. this was not observed for volunteers. interestingly, however, reported levels of attachment were not significantly different between owners and volunteers. for owners, duration of ownership had a positive effect on the level of attachment reported and this effect increased sharply after two plus years of ownership. this contribution to human animal interaction (hai) research suggests that attachment is an important factor in promoting health benefits to owners. http://www.petbehaviourscience.org/ 2011), probably mediated by the oxytocin system (handlin et al. 2011; beetz 2012). pets are usually chosen on the basis of their ability to respond in an affiliative manner (schuelke et al. 1992) and they are perceived as friends which are nonjudgmental, loyal, and fond of their owners (allen 2003; turner et al. 2003; cavanaugh et al. 2008; smolkovic et al. 2012). however, physiological effects on humans from owning a pet can be influenced by psychological, physical, and social measures in a complex way (wood et al. 2005). for example, the level of human-pet attachment has been mostly ignored or under-valued as most studies investigating physiological effects have failed to report it (review in beetz 2012). one exception was the study by vormbrock and grossberg (1988) where attachment towards companion animals was associated with lower mean arterial pressure and systolic blood pressure. attachment is reported more often in studies of mental health benefits because pet owners who are closer to their pets feel less lonely and are more sociable (walsh 2009; herzog 2011). beetz et al. (2012) reviewed 69 studies on hai between 1983 and 2011. several of these studies investigated the effects of hai on the cardiovascular system and reported reductions in blood pressure and heart rate in the presence of a pet such as a dog, particularly during demanding tasks (demello 1999; arhant-sudhir et al. 2011; beetz et al. 2012). other research has shown that petting or talking to a dog is correlated with a significant decrease in blood pressure and heart rate in both owners and pets (allen et al. 1991), as well as better survival rates after myocardial failure (allen et al. 2002; wells 2009; 2011). however, no effects of companion animal have also been found in relation to blood pressure and heart rate measurements as exemplified by straatman et al. (1997) and hansen et al. (1999). only a few studies in the field of hai focused on cats (allen et al. 2002; beetz 2012). however, the cat population in the uk alone is approximately 8.5 million. therefore, it is important to include research on the effects of cats and not just dogs. furthermore, a large portion of the studies were conducted in a laboratory environment. however, such environments are not likely to be conducive to observing normal behaviours. for example, in order to encourage natural behaviour, merola et al. (2015), in his laboratory experiment, used only cats that were accustomed to changes in their living environment. wells (2011) reported some conflicting research results on cats as physiological benefits from interactions were not always present (e.g. friedmann and thomas 1995, reported cat owners more likely to die of a heart attack than non-cat owners), highlighting the need for more studies on this pet species as human-cat relationship are known to be functional and mutually beneficial. for example, cats behaved sensitively to human depressive moods and engaged in more allorubbing of the head and flank (rieger and turner 1999), approached owners who described themselves as feeling numb less often, and approached owners who felt extroverted or agitated more frequently (turner and rieger 2001, review in vitale shreve and udell 2015). cats can also change their behaviour towards an object in line with the emotional message given by the owner (merola et al. 2015) and show a number of attachment behaviours including physical contact, allorubbing, playing, and vocalising with the owner (vitale shreve and udell 2015). in addition, cats have been shown to alleviate negative moods and this effect was comparable to the effect of a human partner (turner et al 2003). health depends on various aspects of a person’s lifestyle: dog ownership is known to be one of them (wood et al. 2005) as this has been linked to more exercise (wells 2011). in addition, the confounding effect of better health associated with dog walking (serpell 1991, wood et al. 2005, wells 2011) is not a factor when studying cats and so studies on cats are more likely to detect any health benefits associated with pet ownership alone. this study contributes to an on-going discussion on hai and is concerned with the effect of levels of attachment between pet and owner as a possible underlying cause of cardiovascular health benefit (winefield et al. 2008; wells 2009) and well-being (crawford et al. 2006). this study focused on whether level of attachment affects physiological measures such as heart rate and blood pressure in the presence and absence of the cat. this study compares the impacts on cat owners with that on volunteers in a cat rescue centre. the attachment bond formed with one’s own pet should promote stronger physiological responses (schuelke et al. 1992; julius et al. 2013). since research has already shown that pet owners gain greater health benefits from their own pets compared to people who use animals as a method of therapy (allen et al. 2002), it is predicted that owners petting their own cat will gain greater short term relaxation, measured through blood pressure and heart rate, than volunteers. among pet owners, it is predicted that higher levels of attachment will correspond with greater decreases in page 2 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 1 12 blood pressure and heart rate. this study contributes to hai research by assessing the effects of cats on human health and thus has implications for healthcare and the promotion of well-being. 2. methods questionnaire the lexington attachment to pets scale (laps, johnson et al. 1992) was used in this study to measure the level of emotional comfort in the owners’ relationships to their cats. this scale consists of 23 items but 2 items (’i think my pet is just a pet’ and ‘i am not very attached to my pet’) were excluded as they were somewhat negative questions and had the potential to cause offence (bps 2014). answers were provided on a 4-point scale (1=strongly disagree to 4=strongly agree; zasloff 1996). therefore possible scores ranged from 23 to 92 with the highest scores representing a higher level of attachment (see appendix 1). participants were given the questionnaire upon acceptance of participation and prior to the beginning of the experiment. measurement of blood pressure and heart rate the omron m2 classic upper arm blood pressure monitor was used to measure blood pressure (systolic and diastolic). pressure was measured in millimetres of mercury (mmhg) and heart rate (hr) in beats per minute (bpm) taken at the upper left arm, before, during, and after the presence of the cat. this was a non-invasive technique and fully automatic. participants participants (age 18+) in this study were recruited across cornwall. cat owners who agreed to participate (n=20) completed a questionnaire, the lexington attachment to pets scale (laps, appendix 1) to establish their level of attachment with their cat. the sample of owners included people aged 18 to 64 (mean=34.58, se=3.39). the same questionnaire was used to assess the level of attachment with volunteers at the cats protection league centre in truro, uk (n=20). the sample included people between the ages of 18 and 65 (mean=41.87, se= 3.86). there was no significant difference between the two groups in terms of age (ttest, t=-1.41, df=38, p=0.16) but to correct for any effect of age on the physiological measurements, age was included in the data analysis (see section 2.6.). none of the participants in this study had previous problems with hypertension, were on any related medication or were smokers. due to participants’ commitments, sessions were run mostly on weekends. experimental setting to provide cat owners and cats with familiar surroundings and to encourage the most natural behaviour from both cat and owner, measurements were taken at the owner’s residence. the living room was the room used in all cases since this is the room where normally the owner and cat most frequently interact. all electronic devices were turned off to avoid disturbances which could have had an effect on the measurements taken. for volunteers, all measurements were taken at the volunteer’s common room in the cats protection league centre in truro, uk, and all measurements for ‘during’ cat presence were carried out in the cat’s pen a few steps away from the common room. procedure upon arrival at each participant’s home, informed consent and the completed questionnaire were collected. the cat was removed from the room for this procedure. in the absence of the cat, the participant was free to rest and sit quietly in the room for a period of 10 minutes to allow for blood pressure and heart rate to stabilize at baseline level. at the end of next 10 minutes (20 min into the session), measurements were taken of blood pressure (systolic and diastolic, mmhg) and heart rate (bpm). the left arm was always used for more accurate measurements. these measurements were called ‘before’ in the data analysis. the cat was then welcomed into the room and no attempt was made to constrain the cat’s movements or actions. most cats did seek attention from the owner, who remained seated. the cat often remained with the owner during the next measurements which were taken after 10 minutes of the cat’s presence (30 min into the session). this measurement was called ‘during’. the cat was then removed from the room by the experimenter and after 10 mins of the cat’s absence (40 min into the session); measurements were taken again and called ‘after’. in all cases, the researcher (fd) sat on the left hand side for ease of measurement and all measurements were taken by fd. upon arrival at the cat rescue centre, informed consent and the completed questionnaire were collected. the volunteer was free to rest and sit quietly in the common room for a period of 10 minutes. blood page 3 cat attachment and human health: owner and volunteer comparison creative common license 4.0 – non commercial – share alike – attribution dinis, f.a.b.s.g. and martins, t.l.f. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science pressure and heart rate measurements were taken at the end of the next 10 minutes (‘before’), on the left arm for accurate results. the volunteer then went to the cat pen a few steps away, rested again for 10 minutes (on a chair placed outside) and then entered the pen. again there was no restriction on the cat’s movements or actions. most cats would show interaction and play with volunteers. after 10 minutes the volunteer would exit the pen, sit on the chair, and blood pressure and heart rate were measured again; this measurement was called ‘during’. the volunteer would return to the common room and after 10 minutes, blood pressure and heart rate would be measured again termed ‘after’). in all cases, fd sat on the left hand side of the volunteer in the common room and outside the cat’s pen and carried out all the measurements. statistical analysis 1. repeated measures data were analysed using the general linear mixed models, gee (repeated measures) procedure in spss (version 21). data on hr, systolic and diastolic pressure were tested for normality and once the distribution was identified the appropriate link function was applied to transform the data. the distribution of the data was identified in all cases as being ‘inversed gaussian’ and therefore ‘log link function’ was used. models included the type of participant (owner or volunteer), time of measurement (before, during and after), the interaction between the two and age of participant as a covariate because age is an important factor in blood pressure (lewington et al. 2002). analyses that were found to be significant were then analysed further with the least significant variables removed using a stepwise deletion process, prior to repeating the analysis. this was repeated for main effects and interactions; however if the interaction was significant no main effects were removed. age remained in the model even if not significant, to control for possible physiological differences due to age (lewington et al. 2002). 2. the laps score: level of attachment the difference between physiological measurements before cat and during cat presence (b-d) was used to assess the effect of level of attachment in both owners and volunteers. the differences (b-d) for hr, systolic and diastolic pressure were tested for normality and data for owners was normal (shapiro-wilk test, all p>0.05) but the data for volunteers was not (all p<0.05). however, the data distribution for level of attachment for owners was identified as not normal (shapiro-wilk test, all p<0.05) and therefore glmms (inversed gaussian, log link) were carried out between b-d differences and level of attachment for both owners and volunteers. in all cases the analyses were carried out using b-d for hr, systolic and diastolic as response and level of attachment as predictor (covariate). however, for volunteer b-d differences, the data had to be made positive by the addition of 15 (as some differences were negative (up to -14) and ‘log link function’ does not accept negative numbers). the figures show the data without this addition. the importance of years of ownership on the level of attachment was explored given that this predictor has been reported elsewhere as being a factor in the laps scores of levels of attachment (cavanaugh et al. 2008). as length of ownership was not normal (shapiro-wilk test, p=0.049), a glmm (inversed gaussian, log-link function) was carried out between level of attachment and time of ownership (years), with the latter defined as a covariate. 3. results physiological measurements heart rate was significantly different over time (fig 1a, 2=27.74, df=2, p<0.001), and it reduced during the cats’ presence in relation to before and after. no significant effect of type of participant was detected (2=3.34, df=1, p=0.07); however the interaction term was significant, showing that owners and volunteers reacted differently to the experiment (2=29.59, df=2, p<0.001), with owners showing a decrease and a corresponding increase in the sequence b-d-a, with volunteers remaining the same. age of participant was not significant (2=2.64, df=1, p=0.10). systolic blood pressure was significantly different over time (fig 1b, 2=27.06, df=2, p<0.001), although participant type was not a significant factor (2=1.04, df=1, p=0.31), showing that both volunteers and owners reacted in the same way. there was no significant interaction term indicating that the decrease was the same in both groups and this term was therefore removed from the model. the covariate participant age, however, was significant (2=16.79, df=1, p<0.001) showing that age was a factor in determining systolic blood pressure. diastolic blood pressure was also significantly different over time and like all other physiological parameters, it reduced during and increased after the pet was page 4 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 1 12 removed from the room (fig 1c, time: 2=10.99, df=2, p<0.001), with owners showing a decrease and a corresponding increase in the sequence b-d-a while the level in volunteers remained the same. for diastolic blood pressure there was no significant difference between owners and volunteers (participant: 2=0.03, df=1, p=0.86) but the interaction term was significant demonstrating that there was a difference in the way owners and volunteers diastolic blood pressure changed over time (interaction term: 2=9.27, df=2, p=0.01). as for systolic pressure, participant age was significant (2=34.02, df=1, p<0.001) indicating that age is also a factor determining diastolic blood pressure. laps scorethe importance of levels of attachment for owners, there was a positive significant effect of level of attachment on the difference in hr between before and during (b-d: 2=21.09, df=1, p<0.001, see fig2a). this means that the reduction in hr with the cat present was positively linked to the level of attachment the owner showed towards the pet. the same significant positive effect of level of attachment was present in both systolic and diastolic blood pressure (systolic b-d: 2=27.45, df=1, p<0.001; diastolic b-d: 2=5.93, df=1, p=0.01, see fig 2b and 2c). for volunteers, reductions in hr, systolic and diastolic blood pressure between before and during the cat’s presence were not affected by volunteers’ level of attachment to cats at the rescue centre (hr b-d, 2=0.25, df=1, p=0.62; systolic b-d, 2=0.00, df=1, p=0.96; diastolic b-d, 2=0.04, df=1, p=0.83). there was no significant difference between owners’ and volunteers’ level of attachment measured by the laps questionnaire (mean score owner=64.95, se=4.96; mean score volunteer=61.90, se=2.07; mann-whitney u=137.5, z=-1.69, p=0.09). level of attachment increased significantly with years of pet ownership for owners (fig. 3; 2=9.40, df=1, p=0.002), showing that the longer owners had the cat, the stronger the laps scores in the questionnaire. 4. discussion consistent with other studies, the results here show that the presence of a cat or petting a cat can reduce heart rate and both systolic and diastolic blood pressure, and that this reduction is generally more pronounced in the cats’ owners rather than in the volunteers from a rehoming centre. for owners, levels of attachment as measured by the laps scale were positively associated with this reduction in metabolic measurements before and during pet presence; i.e. the difference (b-d) was positively correlated with the level of attachment. however, interestingly, reported levels of attachment were not significantly different between owners and volunteers. in addition, duration of ownership had a positive effect on the level of attachment reported by owners and this relationship increased sharply after two plus years of ownership. page 5 cat attachment and human health: owner and volunteer comparison creative common license 4.0 – non commercial – share alike – attribution dinis, f.a.b.s.g. and martins, t.l.f. a c b fig 1. a) heart rate (bpm), b) systolic blood pressure (mmhg) and c) diastolic blood pressure (mmhg) for owners and volunteers before, during, and after exposure to a cat, either their own or from a rehoming centre. for statistical tests see results. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science the reduction in heart rate and blood pressure in the presence of a pet suggests a health effect particularly for owners. some studies have reported significant physiological benefits related to pets while others have not (see review in beetz 2012). benefits have involved reduction in heart rate (demello 1999; handlin et al. 2011) and blood pressure, both arterial and systolic (grossberg and alf 1985; vormbrock and grossberg 1988), and one study has even shown a significant difference between visual, verbal and tactile experience of pets (e.g. dog, vormbrock and gossberf 1988), the latter being significantly more important in reducing blood pressure. the mere presence of a pet, usually a dog, resulted in lower blood pressure and heart rate (allen et al. 2002; wells 2009; beetz et al. 2012). allen et al. (2002) found presence of a pet to be more effective in reducing blood pressure and heart rate than the presence of a partner and friend. the current study adds to the body of evidence that shows these physiological effects but goes further to suggest that these health benefits are related to the reported level of attachment between owner and cat, which is in turn affected by the length of ownership. the experimental design in this study takes into account several criticisms raised in the field since it compares within individuals, in a before, during and after set-up (moody et al. 1996), uses cats rather than dogs (as dogs are related to physical exercise, e.g. friedmann et al. 2003; arhant-sudhir et al. 2011 and are therefore known to have a longer term benefit in cardiovascular disease, wolff and frishman 2005), and considers physiological parameters taken at home or at the work place rather than in a laboratory environment. in addition, the laps attachment scale used here was edited to accommodate the volunteers’ responses (e.g. zasloff 1996) to make it comparable to the owners’ reported attachment levels. other criticisms in this subject area could not be covered in this study (i.e. using people who did not choose to have pets or be around animals, such as in allen et al. 2001), however, we used two sets of people who liked cats, one that owned cats and one that volunteered at a homing centre. the decision to use two sets of people who showed attachment to cats in this study was preferable; page 6 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 1 12 a c b fig 3. relationship between level of attachment (laps score) and years of cat ownership, showing that the level of attachment increases significantly with duration of ownership, particularly after 3 years (2=9.40, df=1, p=0.002). fig 2. laps scores relationship with physiological measurement differences between before and during (b-d) for owners. a) heart rate (bpm), 2=21.09, df=1, p<0.001; b) systolic blood pressure (mmhg), 2=27.45, df=1, p<0.001; and c) diastolic blood pressure (mmhg), 2=5.93, df=1, p=0.01. given that we wanted to carry out the measurements in a familiar set-up and to use people who did not choose to spend time with cats would create a situation where these ‘ordinary’ people would be meeting cats in an artificial set-up (such as in a laboratory environment). research suggests that owners are just as attached to cats as they are to dogs. serpell (1996) assessed owner satisfaction with cats a year after re-homing and found that moderately attached owners were not consistently any less satisfied with pets overall than very attached ones. the importance of ‘affection’ in the cat-owner relationship suggests that simpler behavioural criteria are relevant to cat owner’s attachments (serpell 1996). one possible explanation is that cat behaviour is influenced by characteristics of the human and family, as found by mertens (1995). a body of evidence supports the notion that cats have developed a range of mechanisms in their interaction with humans (review in vitale shreve and udell 2015). even though interactions between cats and owners can be overtly operational (i.e. cats can be trained to go for walks and to follow instructions etc.), the humans and cats used in this study did not have an overtly operational relationship. however, human-cat relationships are surely functional in the social sense, for example; cats in households have been shown to be valuable social companions and social supporters (rieger and turner 1999, wedl et al. 2011). evidence shows that cats are sensitive to human cues such as gaze (merola et al 2015) and comply with humans intent to interact, with resultant less negative moods in their owners (turner et al. 2003). the results in this study do suggest that the functionality of people-cat relationship can extend from effects on mood and perceived social support to provision of physiological benefits. in this study, owners’ physiological responses are compared to volunteers’ and several differences were found. volunteers were willing to give their free time to cats and this commitment to animal care was shown in the lack of difference in their reported level of attachment when compared to cat owners. interestingly, the pet presence effect for volunteers was not strong (even though they also experienced some reduction, particularly in systolic blood pressure) and was not related to the level of reported attachment. it would be expected, however, that levels of self-esteem would be raised by volunteering and increases in selfesteem benefits accrued by a pet have been related to levels of attachment (triebenbacher 1999, crawford et al. 2006) but increases in self-esteem have not yet been linked to health benefits. as volunteers’ systolic blood pressure was also reduced during cat presence, it is important to conduct further research into this effect. owners, as opposed to volunteers, may be more prone to possess personality traits likely to dispose them to enhanced health and well-being (mcnicholas and collins 1998, wells 2011). however this would have to be assessed in a separate experiment comparing owners to non-owners and volunteers. it can be argued that since volunteers are also ‘cat people’, the most likely explanation is that it is the presence of a wellknown pet, and staying at home, that results in positive physiological consequences. the use of the common room in the cat homing centre and the pet holding pen (after resting) when measuring volunteers’ responses should have yielded similar metabolic changes as owners at home, given that laboratory studies have also shown reductions in physiological measurements when in tactile contact with pets (allen et al. 2012; handlin et al. 2011). as measurements on volunteers were always taken after a rest, the different experimental set-up should not have been responsible for the differences between owners and volunteers. the interesting possibility raised by this study is the combination of both ‘family pet and the home environment’ that generates the health benefits, and that tactile contact with a cat that is not ‘very familiar in the work set-up’ yields fewer health benefits. it is important to consider that the effect of a companion animal may be small in comparison to the effect of staying at home (as opposed to going to a shelter), so that the benefit of a companion animal may be additional to the effect of being at home. wells (2009) also reported greater benefit with familiar animals; however, volunteers are familiar with some cats in the centre and develop companion bonds with certain animals. further research could concentrate on responses to tactile contact to pets in different set-ups and using cats with which volunteers have special relationships. in addition, wedl et al. (2011) assessed the interactions between cats and owners and found out that these interactions depended very much on features of the owners not the cat. perhaps not surprisingly, cats have been shown to be neutral pets that comply with the owners’ wishes for some or no interaction (rieger and turner 1999). future studies may concentrate on owners’ and volunteers’ state (such as self-esteem) as well as attachment. the possible effect of familiarity of pet and environment for the owner’s health suggested here is somewhat reflected in the effect of duration of ownership on the levels of attachment reported by page 7 cat attachment and human health: owner and volunteer comparison creative common license 4.0 – non commercial – share alike – attribution dinis, f.a.b.s.g. and martins, t.l.f. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science owners. it suggests that ‘familiarity breeds attachment’ as shown in the positive relationship between the duration of ownership and the level of attachment, and the possible physiological impact. the value of a pet has been shown to increase with length of time of ownership (word 2012) and to be stronger after 3 years (smolkovic et al. 2012); cavanaugh et al. (2008) also reported a relationship between well-being and duration of ownership of dogs. the attachment bond created between owner and pet may have implications for how their relationship is maintained over time (smolkovic et al. 2012). interestingly, in human relationships, satisfaction tends to decline over time (over the first 4 years) and contributes less to overall well-being (kurdek 1998). there has been some interest in the possible mechanisms behind the reported reductions, by many studies, in heart rate and blood pressure. the most likely mechanism is related to cortisol rates mediated by the oxytocin system (beetz 2012). oxytocin reduces glucocorticoid levels in humans (neumann et al. 2000) particularly in relation to social stress (kirsch et al. 2005). oxytocin is known to increase in the blood of both humans and dogs after 5 to 24 minutes of stroking a dog (odental and meintjes 2003). salivary cortisol has been measured in dog owners when either petting their own or an unfamiliar dog, or reading a book quietly; both dog treatments reduced cortisol but reading did not (odendaal 2000; odendaal and meintjes 2003). the increase in oxytocin and reduction in cortisol correlated positively with the quality of the relationship, including physical contact. in humans, oxytocin is also linked with human attachment and bonding (odendaal 2000). in the pet-human situation, the release of oxytocin and reduction in cortisol levels would be expected to be higher in the owner-home combination than in the volunteer-rehoming centre set-up. another suggested mechanism is a decrease in the central autonomic activity as a consequence of positive mood induction (arhant-sudhir et al. 2011), but whichever the possible mechanism, this study suggests that, if positive mood induction is occurring, owners at home are benefiting more than volunteers at a re-home centre. further studies may concentrate on oxytocin in different people-cat-place combinations. 5. conclusions this study suggests that interactions with cats result in positive health benefits for people who enjoy cats. health benefits were larger the stronger the level of attachment between pet and owner, but the familiarity of home and own pet may be an important factor in these health benefits. for rehoming centre volunteers, the health effects of petting cats were smaller. further research may focus on pet-cat-place combinations and on the measurement of oxytocin in addition to blood pressure and heart rate. 6. acknowledgements we gratefully acknowledge the permission granted by libby luckett, manager of the truro cat’s protection league re-homing center and the patience of all participants involved, owners, volunteers and cats. we would also like to thank dr kathy baker from newquay zoo for comments on the statistical analysis and to the following people for comments on earlier versions of this manuscript: roger kayes, rebecca allen, dr peter mcgregor and two anonymous referees. this manuscript would not have been possible without the support and time for scholarly activity generously given to the corresponding author by lawrence moores. the work described here was carried out in accordance with eu directive 2010/63/eu for animal experiments and approval for the study was not necessary since animals used in this experiment were allowed to behave normally and were always in their home or holding pen at a uk cat rehoming centre,. great care was taken during this experiment not to distress the cats in any way. this study has also followed the british psychological association guidelines in seeking consent and making sure the questionnaire and the procedures did not cause offence or lasting psychological harm to the human participants. 7. references allen, k. 2003. are pets a healthy pleasure? the influence of pets on blood pressure. current directions in psychological science 12: 236-239. doi: 10.1046/j.0963-7214.2003.01269.x allen, k. m., blascovich, j. , tomara, j. 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t.l.f. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science individual data for one million adults in 61 prospective studies. the lancet 360: 1903-1013. wos:000179870200007 mcnicholas, j and collins, g.m. 1998. could type a (coronary prone) personality explain the association between pet ownership and health? in: companion animals and human health, ed. c.c. wilson and d.c. turner (eds.). ca: sage. mcnicholas, j., gilbey, a., rennie, a., ahmedzai, s., dono, j.and ormerod, e. 2005. pet ownership and human health: a brief review of evidence and issues. british medical journal 331: 1252-1255. doi: 10.1136/bmj.331.7527.1252 merola, i., lazzaroni, m., marshall-pescini, s., pratoprevide, e. 2015. social referencing and cat-human communication. animal cognition 18:639-648. doi: 10.1007/s10071-014-0832-2 mertens, c. 1995. the human cat relationship. tierarztliche umschau 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animals in an older population. international journal of behavioural medicine 15: 303-310. doi: 10.1080/10705500802365532 wolff, a.i. and frishman, w.h. 2005. animal-assisted therapy in cardiovascular disease. seminars in integrative medicine 2: 131-134. wood. l., giles-corti, b. and bulsara, m. 2005. the pet connection: pets as a conduit for social capital?. social science & medicine 61: 1159-1173. doi: 10.1016/j.socscimed.2005.01.017 word, j. 2012. pet perks: an examination and analysis of the relationship between companion animals and the development of empathy. honors, texas state university-san marcos. texas. zasloff, r. 1996. measuring attachment to companion animals: a dog is not a cat is not a bird. applied animal behaviour science 47: 43-48. doi: 10.1016/01681591(95)01009-2 page 11 cat attachment and human health: owner and volunteer comparison creative common license 4.0 – non commercial – share alike – attribution dinis, f.a.b.s.g. and martins, t.l.f. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science appendix i lexington attachment to pets scale (laps) age group: 18-25 | 25-34 | 35-44 | 45-54 | 55-64 | 65+ female male please state whether you agree or disagree with the following statements about your cat or rescue cat. circle the one which applies best strongly disagree somewhat disagree somewhat agree strongly agree 1. my cat/rescue cat means more to me than any of my friends. 2. quite often i confide in my cat/ rescue cat. 3. i believe that pets should have the same right and privileges as family members. 4. i believe my cat/ rescue cat is my best friend. 5. quite often, my feelings toward people are affected by the way they react to my cat/ rescue cat. 6. i love my cat/ rescue cat because he/she is more loyal to me than most of the people in my life. 7. i enjoy showing other people pictures of my cat/ rescue cat. 9. i love my cat/ rescue cat because it never judges me 10. my cat/ rescue cat knows when i'm feeling bad. 11. i often talk to other people about my cat/ rescue cat. 12. my cat/ rescue cat understands me. 13. i believe that loving my cat/ rescue cat helps me stay healthy. 14. pets deserve as much respect as humans do. 15. my cat/ rescue cat and i have a very close relationship. 16. i would do almost anything to take care of my cat/ rescue cat. 17. i play with my cat/ rescue cat quite often. 18. i consider my cat/ rescue cat to be a great companion. 19. my cat/ rescue cat makes me feel happy. 20. i feel that my cat/ rescue cat is a part of my family. 22. owning a pet / volunteering with pets adds to my happiness. 23. i consider my cat/ rescue cat to be a friend. page 12 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 1 12 oxytocin blocks pet dog (canis familiaris) object choice task performance being predicted by owner-perceived intelligence and owner attachment. jessica lee oliva1, jean-loup rault2, belinda appleton3, alan lill4 pet behaviour science | 2016, vol. 1, 31 -46 jessica lee oliva1, jean-loup rault2, belinda appleton3, alan lill4 1. school of biological sciences, monash university 2. animal welfare science centre, university of melbourne 3. life and environmental sciences, deakin university, geelong 4. school of life sciences, la trobe university, melbourne paper research email: jss_oliva@yahoo.com.au australia. keywords: attachment, bonding, dog, owner, oxytocin highlights this study demonstrates that an owner’s style of attachment to his/her dog may affect both the type of human social cue that the dog can use, and the degree to which it can do so. the study also reveals the importance of past experiences of bonding, with both pets and children, in both the owners’ perceptions of their dog’s cognitive skills, and the style of attachment to it. page 31creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 31 46 abstract a positive association has been found between owner-rated dog cognition and owner-perceived closeness to their dog, using the perceptions of dog intelligence and cognitive skills (podiacs) survey and the monash dog owner relationship scale (mdors). oxytocin has been positively associated with bonding in mammals and with non-verbal intelligence in humans and could therefore explain this relationship between owner-rated questionnaires. the aims of this study were to ascertain: i) whether a pet dog’s performance on an object choice task (oct), which objectively measures dogs’ ability to use human non-verbal, social gestures to find a food reward, could be predicted by their owners’ scores on three different surveys: (a) the mdors, (b) the pet attachment questionnaire (paq), which measures levels of anxious and avoidant attachment styles, and (c) a modified version of the podiacs, and ii) if intranasal administration of oxytocin to dogs, known to enhance dogs’ performance on such tasks, would disable the ability of an owner to predict their dogs’ performance. it was hypothesized that higher ratings of owner-reported closeness to their dog, and higher ratings of owner-perceived intelligence of their dog, would positively predict dog oct performance after saline, but not after oxytocin. seventy-five pet dogs and their owners were recruited to participate in two octs, 5-15 days apart, once after the dog received intranasal oxytocin and once after receiving saline. owners completed the podiacs and another survey relating to pet ownership before oct 1, and the mdors and paq before oct 2. after saline administration, scores on the anxious subscale of the paq were a negative predictor of dog oct performance using pointing cues, while subscale 6 of the podiacs, ‘contagion of human emotions’, positively predicted performance using gazing cues. none of the questionnaire subscales were predictive of performance on the oct after oxytocin administration. results suggest that a dog’s ‘natural’ ability to follow human pointing cues and anxious attachment in owners are inversely related, whilst a dog’s ‘natural’ ability to follow human gazing cues is positively related to ownerrated empathic ability of the dog. http://www.petbehaviourscience.org/ this has implications for training dogs, both for working roles and for pet obedience, which may be complicated by the attachment style of the handler/owner. the study also showed that administration of intranasal oxytocin, which increases dogs’ performance on octs, diminished the relationship between owner attachment and dogs’ oct performance. 1. introduction like human infants, the domestic dog displays attachment behaviours to humans that are absent in even intensively hand-reared wolves (topál et al. 2005), the phylogenetically closest species to the domestic dog (pang et al. 2009). it appears as though the domestication of dogs and subsequent exploitation of them as human companion animals has facilitated the formation of human-dog bonding. one way to evaluate the strength of a human-dog bond is by using questionnaires targeting the dogs’ owners. in 2006, a 28-item questionnaire, the monash dog owner relationship scale (mdors), was developed; it quantifies the ‘perceived emotional closeness’ of the human-dog bond, the degree of ‘dog-owner interactions’, as well as the ‘perceived costs’ (both monetary and to lifestyle) involved in the ownership (dwyer et al. 2006). the strength of the bond, however, does not provide an indication of attachment style. ainsworth et al. (1978) first demonstrated that human infants demonstrate different styles of attachment to their mothers and classified them as either ‘secure’ or ‘insecure’. insecure attachment can be characterised by anxious (anger or rejection of proximity seeking with their mothers after a period of separation) or avoidant (inconsistencies in greeting their mother after a period of separation) behaviours in a strange situation (ainsworth et al.). it has been postulated that internal working models of attachment are developed in infancy and are carried forward into adulthood (for a review see, bretherton and munholland 2008). in adulthood, a secure style of attachment is characterised by a relative ease and comfort in close relationships, with minimal distressing fears of abandonment. an anxious style of attachment is characterised by a desire to be very close in relationships, often to the point which is uncomfortable for the partner, and accompanied by fears of abandonment and inadequacy (brennan et al. 1998). on the other hand, an avoidant style of attachment is characterised by an uncomfortableness in being too close to, or dependent on, others (brennan et al. 1998). recently, zilcha-mano et al. (2011) developed a 26-item questionnaire to evaluate these human, adult internal working models of attachment in relation to pets, the pet attachment questionnaire (paq). in line with literature regarding the different styles of attachment towards other adults (brennan et al.), anxious attachment to a pet involves constant thoughts that something bad might happen to their pet and that they will be left alone, resulting in a constant desire to be near their pet and frustration when their pet does not reciprocate with the same need for closeness (zilchamano et al.). on the other hand, avoidant attachment involves feelings of discomfort when their pet gets too close to them and an avoidance of intimacy with their pet (zilcha-mano et al.). absence of anxious and avoidant attachment types would suggest a secure attachment in which the owner is comfortable with the level of intimacy their pet shows them and is not affected by distressing thoughts or feelings (zilchamano et al.). interestingly, while the paq anxious subscale is moderately correlated with both the anxious and avoidant subscales of the experiences in close relationships scale (ecr), which measures anxious and avoidant attachment in human relationships (brennan et al.), the paq avoidant subscale is only weakly correlated with the ecr anxious subscale. this suggests that people with anxious internal working models of attachment are likely to form an insecure style of attachment to their pet, which is likely to be anxious in nature, but could also be avoidant. on the other hand, people with avoidant internal working models of attachment are also likely to form an insecure attachment to their pets, but which is anxious in nature. this suggests that a pet may allow someone with an avoidant internal working model of attachment to interact with them in a way that they are unable to do with other humans. whilst questionnaires are good tools for quantifying the human-dog bond, they obviously cannot be used on dogs and so, are inherently biased towards the owner’s perspective. currently, there is no biological page 32 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 31 46 marker to assess mammalian bonding; however, many studies have implicated oxytocin as having potential in this role, including in human-dog bonds. in a pioneering study, odendaal and meintjes (2003) found that when humans interacted positively with a dog, gently touching and talking to it, mean arterial blood pressure decreased in both species at the same time serum levels of oxytocin increased significantly, and this was significantly greater than if the human was simply relaxing whilst quietly reading a book. this study employed both men and women, and male and female dogs of differing breeds, but these groupings were not accounted for in the analysis. to account for these potentially confounding variables, miller et al. (2009) replicated odendaal and meintjes’ study using 10 men and 10 women and analysed data for the sexes separately. interestingly, women’s oxytocin secretion significantly increased, whilst men’s secretion level significantly decreased after interaction with their dogs. the reason for this gender dichotomy is unknown; however, it may be due to the influence of sex hormones, known to modulate both the oxytocin peptide and the oxytocin receptor (for a review see, gabor et al. 2012), or differences in the type of interaction between male and female humans, and dogs. for example, although not recorded in miller et al.’s study, calm, soft petting may be more common from women, whilst rough play may be more common from men. indeed, it was demonstrated that plasma cortisol concentrations rose in dogs petted for twenty minutes by men after a venipuncture procedure, but not in dogs petted by women (hennessy et al. 1997), indicating that, contrary to women, men did not stimulate a significant anxiolytic effect in dogs petted after venipuncture. however, when men were trained to pet in a similar fashion to most women, no differences between cortisol levels in dogs petted by men and women were evident (hennessy et al. 1998). there was, however, considerable individual variability in serum oxytocin levels among the men and among the women in miller et al.’s study. in addition, one woman and one man appeared as outliers, with values two standard deviations greater than the mean. the authors noted that the aberrant woman had the greatest number of pet dogs of all participants in the study, whilst the man was the only participant with small children. this might suggest that their oxytocinergic systems had been primed by past bonding experiences. however, caution should be applied when interpreting these findings because while odendaal and meintjes used established chromatographic methods to measure oxytocin, miller et al. employed enzyme immunoassay methods without prior serum extraction, which has recently been shown to be necessary in obtaining valid results in plasma (christensen et al. 2014; szeto et al. 2011). miller et al. also obtained values much higher than the normal biological range yielded using validated methods (christensen et al.; mccullough et al. 2013). it is also important to note that both of the abovementioned studies measured peripheral oxytocin concentrations, which may not reflect central oxytocin function as the peptide does not readily pass the bloodbrain barrier (robinson 1983; veening et al. 2010) and can be independently released in the brain and in the body (amico et al. 1990; engelmann et al. 2000; ludwig and leng 2006; robinson and jones 1982). despite this, peripheral measures of oxytocin concentrations have continued to be the most widely investigated neurochemical metric in human-dog bonding studies (nagasawa et al. 2009; nagasawa et al. 2015; handlin et al. 2011; mitsui et al. 2011). keeping gender and dog breed constant by only including female humans and male labrador dogs, handlin et al. found that plasma oxytocin levels significantly increased in both the dog and female owner after just three minutes of petting, stroking and talking to the dog. however, this study’s findings also need to be interpreted with caution as they too carried out enzyme immunoassays using unextracted plasma samples, and obtained unnaturally high levels of oxytocin. in addition, when analysing grouped data, it is difficult to conclude which dogs, if any, are more bonded with their owners. to investigate this, nagasawa et al. separated participants into two distinct groups, one which expressed a significantly higher perceived degree of satisfaction and communication with their dogs than the other group. after 30 minutes of interaction with their dogs, members of the former group excreted significantly higher concentrations of oxytocin in their urine than members of the latter group, and owned dogs that gazed at them for longer periods within the 30 minute interaction. this was only true, however, for interactions where owners were instructed to look back at their dogs, and not for interactions where they were page 33 oxytocin blocks owner perceptions predicting dog object choice task performance creative common license 4.0 – non commercial – share alike – attribution oliva et al. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science instructed not to look back at their dogs. urine is not yet a validated method for measuring oxytocin (mccullough et al. 2013), however, this may support the role of oxytocin as at least one of the neuropeptides involved in human-dog bonding, and also of gaze as an important factor, similar to the situation observed in human infants (striano et al. 2006; dickstein et al. 1984). the importance of gaze was further reinforced in a similar study which demonstrated that mutual dogowner gaze increased oxytocin in the owner’s urine, and intranasally administering oxytocin increased gazing behaviour in female dogs, which subsequently increased their owners’ urinary oxytocin levels (nagasawa et al. 2015). intranasal administration of neuropeptides is a popular method to study their central effects because, despite a poor understanding of their mechanisms of action (leng and lugwig 2016), increased levels of neuropeptides can be found in brain dialysates (neumann et al. 2013) and cerebrospinal fluid (csf) (born et al. 2002; dal monte et al. 2014; rault 2016; striepens et al. 2013) following administration. in humans, intranasally administered oxytocin increases the salience of social cues (see review by bartz et al. 2011), as well as causing an increase in gaze towards the eye region of other human faces (guastella et al. 2008) (also observed in monkeys (dal monte et al. 2014)). furthermore, variations in the oxytocin receptor gene have been associated with human infant attachment (chen et al. 2011) and interact with internal working models of attachment in adulthood in assessing risk and in feelings of closeness (denes 2015). variations in this gene have also been associated with non-verbal intelligence (lucht et al. 2009), amygdala activation during the processing of emotionally-salient human faces (tost et al. 2010), performance on the “reading the mind in the eyes” test (rodrigues et al. 2009) and autism (wu et al. 2005), a syndrome characterized by social deficits (for a review see, volkmar 2011). in dogs variations in the oxytocin receptor gene have been associated with proximity seeking towards humans and friendliness towards strangers (kis et al. 2014). intranasal oxytocin in dogs has been shown to increase positive expectations (kis et al. 2015), decrease friendliness in response to a threatening person (hernádi et al. 2015), and increase mutual gaze with their owners (nagasawa et al. 2015). it has also been shown to influence dogs’ ability to perform an object choice task (oct) which tests their ability to use human social cues to find hidden food treats (oliva et al. 2015). data from this study will be drawn on in the current investigation of the relationship between the dog owners’ perceptions of the strength of the bond with their dog and of the dog’s intelligence, and the dog’s actual performance level on the oct (see table 1 for relevant means and standard deviations). recently a survey was developed that aimed to determine owner-perceived cognitive abilities in dogs (in general), the perceptions of dog intelligence and cognitive skills (podiacs) survey (howell et al. 2013). the survey measures owner-perceived abilities across eight cognitive domains: ‘recognition of human emotions’, ‘learned problem-solving abilities’, ‘instinctive awareness of human attention’, ‘learned awareness of human attention’, ‘deception’, ‘contagion of human emotions’, ‘instinctive problem-solving abilities’ and ‘general intelligence compared to humans’. while each cognitive domain was significantly correlated with the second subscale of the mdors, the owner’s ‘perceived emotional closeness’ with his/her dog (howell et al.), it is currently unknown if either of these variables relates to actual cognitive ability. if central oxytocin facilitates i) bonding of a dog to a human and ii) the ability of the dog to use human social cues, we would expect to find a positive correlation between owner-rated measures of bonding and social cognition, and actual ability of the dogs to perform well on a task that tests their ability to use human social cues. hence, the first aim of our study was to elucidate whether dog owners’ scores on the mdors, paq and podiacs were related to the dogs’ actual performance on an oct. in particular, we expected to find positive associations between the mdors subscale 2, which measures the ‘perceived emotional closeness’ of the bond, and the podiacs subscales 3 or 4, which contain questions appropriate to the oct, such as “my dog can instinctively understand human gestures like pointing at food or toys” (subscale 3) and “my dog can learn to understand human gestures like pointing at food or toys” (subscale 4), and oct scores. the influence of attachment style page 34 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 31 46 (known to be affected by the oxytocin receptor gene (chen et al. 2011; denes 2015)) and other factors relating to the dog and owner that may influence oxytocin function (such as parental and pet ownership history (miller et al. 2009) and order of intranasal treatments (oliva et al. 2015)) were also explored. the second aim of the study was to investigate the effect of intransally administering dogs with oxytocin on this predictive relationship. due to the fact the oxytocin enhances dog performance on octs, we expected this predictive relationship to be diminished as dogs with owners scoring low on the mdors subscale 2 and the podiacs subscales 3 and 4 may be performing the task equally as well as dogs with owners who score highly on these subscales, after oxytocin administration. 2. methods study animals seventy-five pet dogs (33 males, 42 females) and their owners (14 males, 48 females) were recruited for the study. owners were recruited voluntarily through poster advertisements at monash university’s caulfield and clayton campuses, as well as through university enewsletters and social media websites. the dataset of behavioural performance is the same as published in oliva (2015). the study was approved by the monash university school of biological sciences animal ethics committee (bsci/2013/07) and the monash university human research ethics committee (cf12/08472012000385). materials 0.5 ml saline solutions containing oxytocin (24 iu) or saline only, were administered to the nostrils of the dogs using a mucosal atomizer device connected to a 1ml syringe. four identical, opaque spaniel bowls were used to conceal the food treats and oct scores (out of 20 per cue) were recorded by the experimenter using pen and paper. a modified version of the podiacs (specifically about the owner’s dog, as opposed to dogs in general), and another questionnaire about other factors relating to the dog and owner that may influence oxytocin function were administered to participants before the first oct (refer to the appendix). each cognitive domain of the podiacs has been previously found to have good internal consistency, with cronbach alpha values (a co-efficient of internal consistency commonly used as an estimate of reliability) of 0.74-0.91 (howell et al. 2013). the mdors and paq were administered before the second oct, and completed by participants using pen and paper. the mdors scale has been found to have good content validity and good internal consistency, with cronbach alpha values of 0.84 for both ‘perceived emotional closeness’ (subscale 2) and ‘perceived costs’ (subscale 3) (dwyer et al. 2006). the ‘dog-owner interaction’ (subscale 1) co-efficient was not as strong, with an alpha value of 0.67 (dwyer et al.). the paqs questionnaire has also been found to have good content and construct validity, good test-retest reliability (r=.75) and good internal consistency, with cronbach alpha values for avoidant attachment between 0.84 and 0.91 and for anxious attachment between 0.86 and 0.92, with a weak, non-significant correlation between the two (r = 0.1 to 0.4) (zilcha-mano et al. 2011). procedure dogs were pseudo-randomly separated into two groups: those that received oxytocin in their first session and saline in their second session (oxy-sal) and vice versa (sal-oxy). upon arrival at the testing room in session 1, the dogs received one of two intranasal treatments, oxytocin or saline. when dogs arrived for their second session, they received the other intranasal treatment. there was a 5-15 days washout period between sessions. both the experimenter and the dog owner were ‘blind’ to which treatment the dog received on which day, as solutions were labelled ‘a’ or ‘b’ and only decoded after the study concluded. regardless of which group the dogs were allocated to for the experimental study, owners of all dogs completed the questionnaires in the same order mentioned above, while the dog was free to roam the testing room, interact with the owner and experimenter, or wander outside. the questionnaires were completed in this order so as not to influence the owner’s opinion about their dogs’ ability to perform the task before completion of the podiacs. forty-five minutes after the treatment was administered, the oct commenced, which involved (a) pre-training (before each 10 trial block of the oct proper): four correct trials in a row where the dog was shown a food treat being dropped page 35 oxytocin blocks owner perceptions predicting dog object choice task performance creative common license 4.0 – non commercial – share alike – attribution oliva et al. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science into one of two dog bowls, either side of the experimenter (delivered using the ipsilateral hand from a kneeling position, 75cm from the target) (b) 20 trials (block 1 and block 3) where the dog was given a momentary distal pointing cue (a 1-2 second point delivered using the ipsilateral index finger from the same kneeling position) to indicate the location of the hidden food reward located in one of the two dog bowls and (c) 20 trials (block 2 and block 4) using a gaze cue (a 1-2 second gaze shift delivered from the same kneeling position, keeping the head straight). five control trials per block, where no cue was given, were also delivered to ensure the dogs were not relying on scent to solve the task. the task usually lasted between 45 and 60 minutes and was carried out by the experimenter with the help of the owner, who called the dog back to the starting position between trials and restrained it until the experimenter said the release word “ok”. the experimenter recorded the score for each trial as either correct (if the dog approached the bowl with the hidden food treat) or incorrect (if the dog approached the empty bowl or the experimenter) before starting the next trial. trials were also considered incorrect if the dog made no choice but was deemed motivated to complete the task by participating when two of the easier pre-training cues were then given as a test of motivation. data pertaining to oct performance is the same as that which was published in oliva (2015) and means and standard deviations pertaining to this data are reported in table 1. data analysis the raw scores (out of 20) were calculated for each cue used in the oct proper after oxytocin and saline administration and entered into ibm spss statistics version 22 (spss ibm, new york, u.s.a., 2013). totals for each subscale of the podiacs, mdors and paq were calculated and divided by the number of items in that subscale to obtain overall mean score values. some items were reverse-scored according to the scoring instructions for the particular questionnaire. all mdors subscales were scored such that higher values reflect greater relationship quality, so higher values on mdors subscale 3 ‘perceived costs’ actually represent lower perceived costs. cronbach alpha values were calculated to obtain an indication of internal consistency for each of the subscales used in the current sample. pearson’s correlations were run between the cognition and the attachment subscales to see if previous findings of an association between owner-reported bonding/attachment and owner-perceived cognition ratings (howell et al. 2013) were replicated in our study, and to investigate the effect of attachment style. multiple regression was employed to test the predictive power of other factors relating to the dog, and ownership of the dog, in response to each subscale. to test whether dogs’ performance on the oct could be predicted by i) their owners’ questionnaire data, ii) other factors relating to ownership, and iii) gender of the dog and order of treatment administration, hierarchical multiple regression was run for each of the outcome variables: point oxytocin, point saline, gaze oxytocin and gaze saline. dummy variables were created for the ‘other factors’ predictors: previous dog owner, previous non-dog owner (owners with a history of owning pets other than dogs), owner parental history, young dog, old dog, gender of owner, entire dog, inside dog. dummy variables were also created for order of treatment administration and gender of dog. each subscale of the podiacs, mdors and paq was entered at step 1 of the regression; ‘other factors’ were entered at step 2 and gender and order of treatment administration were entered at step 3. preliminary analyses were conducted for each of the regressions to ensure that there was no violation of the assumptions of normality, linearity, multicollinearity and homoscedasticity. an alpha level of 0.05 was selected for all tests of significance. page 36 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 31 46 table 1. mean (± standard deviation) correct object choices out of 20 for all dogs combined, according to treatment. 3. results chronbach alpha values for each subscale are given in table 2. sample means and standard deviations for each of the subscales are given in table 3. pearson’s correlations between each of the perceived intelligence subscales and each of the attachment subscales are given in table 4. as evident from table 4, only subscale 8 of the podiacs was positively associated with the mdors subscale 2, subscales 3, 5, 8 were positively associated with the mdors subscale 1 and the podiacs subscale 8 was positively associated with the mdors subscale 3. furthermore, we found a positive association between subscale 6 of the podiacs and paq anxious subscale. to test whether any ‘other factors’ relating to the dog and ownership of the dog contributed to questionnaire scores, multiple regressions were run for each subscale of each questionnaire with ‘other factors’ as predictor variables. none of the factors were successful in predicting scores for the podiacs subscales 1-5, and 8, the mdors subscales 1-3 nor the paq anxious subscale. however, ‘previous dog owner’ negatively influenced podiacs 6 scores (contagion of human emotions), whilst parental history positively influenced podiacs 6 scores, with 20% of the variance being explained by the model: f8, 66 = 2.11; p=.047. male owners and previous non-dog owners positively influenced podiacs 7 scores (instinctive problemsolving abilities), with 23% of the variance being explained by the model: f8, 66 = 2.49; p=.020. parental history positively influenced paq avoidant scores, with 23% of the variance being explained by the model: f8, 66 = 2.52; p=.019. page 37 oxytocin blocks owner perceptions predicting dog object choice task performance creative common license 4.0 – non commercial – share alike – attribution oliva et al. table 2. cronbach alpha values for each subscale of the podiacs, mdors and paq, compared with values previously found in the literature. n = 75 https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science regression co-efficients for the above-mentioned significant models can be found in table 5. of the 75 dogs involved in the study, two males and 11 females did not complete the two sessions of the experimental study successfully, leaving a total of 31 males and 31 females in the following hierarchical multiple regression analysis. none of the models were able to predict scores for pointing or gazing after oxytocin administration. for pointing scores, the subscale scores entered at step 1 explained an insignificant 18% of the variance: f13, 49 = 0.84; p=.62. the ‘other factors’ were entered at step 2 but did not contribute significantly to the model: r2 change = .18; f change 21, 41 = 1.39; p=.23. additionally, ‘oxytocin administered first’ and ‘female’ were entered at step 3, but they too did not significantly contribute to the model: r2 change =.01; f change 23, 39 = 0.23; p=.79. for gazing scores, the subscale scores entered at step 1 explained an almost significant 35% of the variance: f13, 48 = 1.93; p=.050 (with only the podiacs subscale 4 – learned awareness of human attention – negatively predicting performance). once again, the ‘other factors’ entered at step 2 did not contribute significantly to the model: r2 change = .09; f change 21, 40 = 0.74; p=.66, and ‘oxytocin administered first’ and ‘female’ also didn’t significantly contribute to the model when entered at step 3: r2 change =.02; f change 23, 38 = 0.76; p=.48. in contrast, model 3 was successful in predicting pointing scores after saline administration and model 1 was successful in predicting gazing scores after saline administration. for pointing scores, the subscale scores entered at step 1 explained an insignificant 29% of the variance: f13, 50 = 1.57; p=.13. the ‘other factors’ entered at step 2 did not contribute significantly to the model: r2 change = .07; f change 21, 42 = 0.58; p=.79. page 38 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 31 46 table 3. descriptive statistics for owners’ scores on each subscale of the podiacs, mdors and paq. n = 75 however, when ‘oxytocin administered first’ and ‘female’ were entered at step 3, they did contribute significantly to the model: r2 change =.22; f change 23, 40 = 10.44; p<.0001. for gazing scores, the subscale scores entered at step 1 explained a significant 35% of the variance: f13, 49 = 2.07; p=.034. the ‘other factors’ entered at step 2 did not significantly contribute more to the model: r2 change = .14; f change 21, 41 = 1.41; p=.22, nor did ‘oxytocin administered first’ and ‘female’ when entered at step 3: r2 change =.02; f change 23, 39 = 0.77; p=.47. regression coefficients for the above-mentioned significant models are given in table 6. as evident from table 6, the paq anxious subscale was a significant negative predictor of performance on the oct with pointing cues after saline administration; indeed, dogs owned by people who score one point higher on this subscale than other dog owners would be expected to score 1.49 points lower on the task than dogs owned by people who score one point lower on this subscale. scores were also likely to be lower for female dogs and higher for dogs that received oxytocin on their first visit (thus making pointing after saline scores their pointing scores for their second visit). in contrast, the podiacs 6 subscale (contagion of human emotions) was a significant positive predictor of performance on the oct with gazing cues after saline administration whereby dogs owned by people who score one point higher on this subscale than other dog owners would be expected to score 1.72 points higher on the task than dogs owned by people who score one point lower on this subscale. page 39 oxytocin blocks owner perceptions predicting dog object choice task performance creative common license 4.0 – non commercial – share alike – attribution oliva et al. table 4. pearson correlation matrix describing relationships between the 8 podiacs subscales and the 3 mdors and 2 paq subscales. n = 75 https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science page 40 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 31 46 table 5. unstandardized (b) and standardized (β) regression coefficients for each predictor variable in a hierarchical multiple regression for podiacs 6, podiacs 7 and paq avoidant scores. 4. discussion contrary to our hypothesis, no significant, positive correlation was obtained between the mdors 2 (perceived emotion closeness) and the podiacs 3 (instinctive awareness of human attention) or 4 (learned awareness of human attention). none of these subscales predicted oct scores (after saline nor oxytocin) either, although, interestingly, podiacs 4 was shown to negatively predict gaze scores after oxytocin, however this model did not quite reach significant overall. there was, however, a significant, positive correlation between the mdors 2 and the podiacs 8 (general intelligence compared to humans). in addition, the paq anxious subscale negatively predicted oct scores (as well as being female and receiving oxytocin second) using pointing cues and the podiacs 6 predicted oct scores using gazing cues. in contrast to howell et al.’s (2013) study, which showed a positive association between mdors subscale 2 and all 8 subscales of the podiacs, mdors subscale 2 was correlated with only one of the eight cognitive domains assessed in the podiacs, subscale 8. the discrepancies between our findings and previous results are possibly due to the fact that whilst our modified version of the podiacs assessed perceived intelligence and cognitive skills of owned dogs, howell et al.’s original survey assessed perceived intelligence and cognitive skills of dogs in general. they may also be due to the differences in internal consistency, refer to table 2, particularly the podiacs subscale 3 which resulted in a questionable cronbach alpha value of .64 in the current study but a much more acceptable value of .80 in howell et al.’s study. for other subscales, such as 7 and 8, cronbach alpha values in the current study are much higher than in howell et al’s study, however, howell et al.’s study also had a much larger cohort of participants, so it is possible the discrepant findings are due to the current study having lower statistical power. the podiacs subscale 8 was also associated with subscale 3 of the mdors (perceived costs), which is consistent with howell et al.’s findings, while the mdors subscale 1 (dog-owner interactions), was associated with subscales 3: (instinctive awareness of human attention), 5: (deception) and 8, in the current study (and all but subscale 7 in the study by howell et al.). taken together, these findings suggest that time spent with one’s dog is more important than how psychologically close one feels towards their dog in perception of intelligence and only minimally support our hypothesis that owners who are more closely bonded to their dog will perceive him/her to be more page 41 oxytocin blocks owner perceptions predicting dog object choice task performance creative common license 4.0 – non commercial – share alike – attribution oliva et al. table 6. unstandardized (b) and standardized (β) regression coefficients for each predictor variable in model 3 (mdp cues) and model 1 (gaze cues) of a hierarchical multiple regression for point and gaze cue scores after saline administration on the object choice task. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science intelligent. additively, we found a positive association between subscale 6 of the podiacs, (contagion of human emotions), and the paq anxious subscale. this may suggest that anxiously-attached owners believe more than non-anxiously attached owners that their dog can feel sad, afraid, angry and/or happy when its owner feels the same emotion. this may be a misperception, due to having higher levels of anxiety, or the result of these owners expressing more emotions, particularly negative ones, which their dog finds distressing and to which it reacts more overtly. however, our results suggest that dogs of owners who score highly on this subscale are more likely to perform more poorly on the oct using pointing cues, in the absence of exogenous oxytocin administration. o’farrell (1995, as cited in o'farrell 1997) found a significant correlation between owner anxiety and over-excitement and displacement activities in the dog, hence, this poorer performance may be a direct effect of the anxiously attached owner who was present in the room at all times. when ‘other factors’ relating to ownership were considered, it was found that podiacs 6 scores (contagion of human emotions) were negatively influenced by the factor ‘previous dog owner’, whilst ‘parental history’ positively influenced these scores, suggesting that first time dog owners and owners who have had children are more prone to believe that their dogs feel their emotions more than experienced dog owners and owners who have not had children. ‘male owners’ and ‘previous non-dog owners’ positively influenced podiacs 7 scores (instinctive problemsolving abilities). this may highlight differences in interactions between male and female owners and their dogs (hennessy et al. 1998, hennessy et al. 1997) and also suggests that past experiences of pet ownership may affect the perception of current dog ownership, whereby experienced pet owners believe their dogs to have more instinctive problem-solving abilities. furthermore, ‘parental history’ positively influenced paq avoidant scores, which suggests that owners who have had children are more likely to form avoidant type attachments to their dog, than owners who have not had children. according to zilcha-mano et al. (2011), there is no correlation between avoidant styles of attachment towards humans and towards pets. therefore, it is possible that due to the parent-offspring relationship(s) in their lives, the relationship to their pet is relatively low in priority and this is reflected as an avoidant owner-pet relationship. performance on the oct after oxytocin administration could not be predicted by any of the statistical models. this is likely to be due to the fact that dogs’ abilities to perform octs correctly are enhanced by oxytocin administration (oliva et al. 2015) and therefore differs from what the owner would otherwise expect. while oliva et al. were able to demonstrate an increase in dogs’ ability to correctly use human social cues following oxytocin administration, it remains unknown why this is so. it may reflect an effect of oxytocin in increasing a dogs’ ability to understand human gestures, or, in light of the findings relating to the paq anxious subscale and poorer performance on the oct using pointing cues after saline, it could that mean that oxytocin, also known also for its anxiolytic properties (de oliveira et al. 2012; heinrichs et al. 2003), reduces the anxiety these dogs may ‘absorb’ from their owner, thereby disinhibiting their natural cognitive ability. oxytocin’s performance enhancing effects may also may explain why the podiacs 4 was a negative predictor of performance, rather than a positive one as was hypothesized, in the almost significant multiple regression for gaze after oxytocin administration i.e. if the owners rated their dogs low for ‘learned awareness of human attention’ and the dogs performed better than expected due to oxytocin. in contrast to the above-mentioned lack of predicative power following oxytocin administration, performance on the oct after saline administration, could be predicted by the models. as previously mentioned, paq anxious scores negatively predicted oct scores for pointing cues; thus, dogs owned by people who score higher on this subscale would be expected to perform more poorly on the task than dogs owned by people who score lower on this subscale. in addition, point scores after saline administration were likely to be lower for female dogs, suggesting that male dogs are more skilled at the oct task, and higher for dogs performing the task for the second time (i.e. their second testing session). findings in oliva et al. (2015) suggest that this is a reflection of the enhanced performance from the oxy-sal group dogs being page 42 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 31 46 maintained in session 2. performance using gazing cues was predictable from scores on the podiacs 6 subscale (contagion of human emotions) after saline administration i.e. dogs owned by people who score higher on this subscale would be expected to score higher on the task than dogs owned by people who score lower on this subscale. this is interesting because this subscale was also associated with the paq anxious subscale, which was negatively influenced by previously owning a dog and positively influenced by parental history. this suggests that past experiences of owning a dog or parenting a child may not only affect serum oxytocin levels (miller et al. 2009), but perceptions about the current dog-owner relationship as well. this finding suggests that gaze is important in human-dog communication for dogs with owners who are anxiously attached to them and hence, potentially more likely to express their emotions to their dog, who may respond accordingly. 6. conclusions overall, this study found little evidence supporting an association between owner-perceived closeness with their dog, owner perceptions of the intelligence of their dog and the actual intelligence of their dog as manifested in the oct (performed after saline and oxytocin administration). the investigation did, however, reveal that past experiences of bonding, with both pets and children, affected owners’ perceptions of the cognitive skills of the currently-owned dog and the style of attachment to it. higher scores for anxious attachment to dogs significantly predicted lower scores on the oct following saline administration using pointing cues. an association was also found between anxious attachment and perceived ‘contagion of human emotions’, which significantly predicted higher scores on the oct following saline administration using gazing cues. this suggests that communicative signals delivered to dogs may differ between anxious and nonanxiously attached owners, affecting the human cues the dog learns to follow. alternatively, communicative signals may be interpreted differently by dogs owned by anxiously-attached humans versus those owned by non-anxiously attached humans. as such, training dogs, both for working roles and for pet obedience, may be further complicated by the attachment style of the handler/owner. 7. acknowledgements we would like to thank the clear dog shop and love’ em for sponsoring the study and providing dog food rewards. we would also like to thank lachlan macquarie, department of econometrics and business statistics, monash university, for his statistical advice. 8. references ainsworth m.d.s., blehar m.c., waters e., and wall s. eds. 1978. patterns of attachment: a psychological study of the strange situation. 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behavioral effects; a review. physiology and behavior 101(2): 193-210. volkmar f.r. 2011. understanding the social brain in autism. developmental psychobiology 53(5): 428-434. wu s., jia m., ruan y., liu j., guo y., shuang m., gong x., zhang y., yang x., and zhang d. 2005. positive association of the oxytocin receptor gene (oxtr) with autism in the chinese han population. biological psychiatry 58(1): 74-77. zilcha-mano s., mikulincer m., and shaver p.r. 2011. an attachment perspective on human-pet relationships: conceptualization and assessment of pet attachment orientations. journal of research in personality 45: 345357. page 46 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.1 | 31 46 influence of dog presence on the tolerance and evaluation of aversive stimulation evaluating fido: developing and pilot testing the field instantaneous dog observation tool amy e. bauer, mary jordan, monica colon, traci shreyer, and candace c. croney pet behaviour science | 2017, vol.4, 1 – 15 doi: 10.21071/pbs.v0i4.5766 amy e. bauer, mary jordan, monica colon, traci shreyer, and candace c. croney 1. department of comparative pathobiology purdue university college of veterinary medicine usa 2. purdue university college of veterinary medicine usa 3. university of puerto rico at mayaguez. puerto rico. paper research * email: ccroney@purdue.edu united states keywords: dogs; commercial breeding; behavior; welfare; assessment highlights • the field instantaneous dog observation (fido) tool, a non-invasive, field ready tool designed to capture the immediate welfare status of dogs housed in commercial breeding facilities was developed and pilot tested for inter-rater reliability. • affiliative or neutral (green) responses to approach were most commonly identified by both novice and expert raters in this study. • both expert and novice raters achieved substantial levels of agreement when using the behavioral component of the fido tool. • there was no statistically significant effect of caretaker presence on response to approach in this study. page 1creative common license 4.0 – non commercial – share alike – attribution 2017 | vol. 4 | 1 15 abstract field assessments of the health and welfare of kenneled dogs (canis lupus familiaris) must be both accurate and rapid. in order to facilitate such evaluations, especially by individuals with limited training in canine behavior and welfare, a non-invasive tool was developed and pilot-tested utilizing dogs housed in commercial breeding facilities. behavioral responses to approach were organized into three categories: red, indicating a fearful response to approach, green, indicating an affiliative or neutral response to approach, and yellow, indicating an ambivalent response to approach. the inter-rater reliability (irr) of the tool when used by both behavioral experts and novice raters was evaluated with and without the presence of the dog’s familiar caretaker. utilizing cohen’s kappa, the experts had almost perfect levels of agreement (kappa=0.87). the novice raters had substantial levels of agreement (kappa=0.74). overall, the dogs assessed by the novice raters had high proportions of green responses to approach and there was no statistically significant effect of caretaker presence on the proportion of green responses to approach. the assessment tool evaluated herein appears to have a high degree of irr whether used by experts in canine behavior or by novice raters and may be a useful screening tool to determine the need for more in-depth welfare assessments. http://www.petbehaviourscience.org/ 1. introduction accurate assessments of dog behavior and welfare help to determine both how well individual dogs cope with their environments and the adequacy of those environments. these evaluations are especially important in facilities where dogs are housed for extended periods of time such as commercial breeding kennels. dogs in high volume breeding facilities are widely believed to live in environments lacking in enrichment and social contact with their caretakers (mcmillan et al. 2011). stress associated with these environmental inadequacies can affect physical, behavioral, and emotional health for both the breeding dogs and their puppies (mcmillan et al. 2011). achieving the goal of providing an ethical source for companion dogs (bir et al. 2016) depends in part on a reliable welfare assessment tool that can be utilized in breeding and other kennel settings from which dogs may be acquired. the welfare of an animal is defined as its ability to cope with its environment (broom 1991) and is both an individual metric and one that changes over the animal’s lifetime. welfare has physical and behavioral manifestations (broom 1991) and both aspects must be included in an assessment tool. however, because of concerns about observer subjectivity and situational variation in animal responses, behavioral metrics of welfare are often minimal or absent as components of welfare assessment tools. an example of situational variation is play behavior, which is a good indicator of positive well-being (boissy et al. 2007; vinke et al. 2005). however, failure to observe play during an assessment does not necessarily indicate poor welfare, particularly if a different behavior, such as eating, was being performed at the time of observation. nevertheless, when validated and tested for reliability, many behaviors can be excellent proxy metrics of animal mental health and well-being. along these lines, assessments of fear have been included as key behavioral indices in evaluations of welfare in production species (muri et al. 2013; powell et al. 2016). likewise, fear in dogs has been used as an indicator of their early rearing experiences that has implications for their overall quality of life. for example, dogs with little human contact during the socialization window have been reported to have a lasting, generalized fear of people, resulting in related deficits that also significantly impact their puppies (serpell and jagoe 1995; lindsay 2000). incorporation of behavior into welfare assessments is therefore essential to fully understand how well animals are coping in a given environment. animal-based assessments often focus on ethograms, lists of specifically defined behaviors associated with both negative and positive emotional states (kiddie and collins 2014; protopopova et al. 2014). challenges with ethogram-type assessments include observer drift (interpretation of behavior changing over time with increased familiarity) (kiddie and collins 2014; protopopova 2016), fixation on specific behaviors that results in improper interpretation of the animal’s overall state (vas et al. 2008), and limited time for assessment of each specific behavior in the field setting. however, these challenges can be addressed via validation of the behaviors of interest as well as robust observer training. in addition, a benefit of ethogramtype assessments is their detail, which allows the prevalence of and the ability of raters to identify each behavior to be determined. behaviors that are rare or difficult to interpret consistently can be identified and either removed from the assessment or focused on in training. a rapid, non-invasive, field ready welfare assessment tool that is reliable is greatly needed for commercial breeding facilities as it could be utilized by researchers, facility inspectors, and staff interested in promoting high welfare standards for dogs. among the concerns reported about dogs in such kennels is heightened fearfulness when confronted with unfamiliar people that has been noted in dogs originating from commercial breeding facilities (mcmillan et al. 2011; mcmillan 2017). however, to our knowledge, no studies have been conducted in the commercial breeding environments in which the dogs are kept. a tool that permits objective evaluations of dogs in these types of kennels would facilitate improved understanding of the quality of their early rearing environments relative to the development of behaviors observed later. page 2 www pet behaviour science org creative commons license 4.0 – non commercial – share alike – attribution 2017 | vol.3 | 1 15 the first objective of this study, therefore, was to develop a rapid, non-invasive, field ready assessment tool useable by individuals without extensive training in canine behavior. the second objective was to pilot test the field readiness of the newly created field instantaneous dog observation (fido) tool, and determine inter-rater reliability (irr). it was important to do this with experts as well as with raters with limited training in canine behavior (novice raters) to ensure broad usefulness of the tool. while dogs kept in commercial breeding facilities have not been well studied, the responses of dogs to unfamiliar people have been found to differ based on the presence of a familiar person (pullen et al. 2012; kis et al. 2014). thus, a final objective was to evaluate the dogs’ responses to approach both with and without their caretakers present. 2. methods facilities all observations were conducted in united states department of agriculture (usda) licensed commercial breeding facilities. kennel size varied by facility to meet or exceed the usda requirements for dogs (usda, 2017). all dogs were provided unrestricted access to an outdoor run during the day. statement of approval the procedures described in this manuscript were reviewed and approved by the purdue university institutional animal care and use committee. initial tool development the assessment tool developed in this study includes measurements of physical health and behavioral wellbeing. the physical health variables are described in table 1 (hurt, 2016). the behavioral component of the tool utilizes response to approach by a stranger as a surrogate measure of socialization toward people. an ethogram consisting of 43 different variables, including the dog’s position in the kennel when initially approached by a stranger (table 2) was developed for test use based on established indicators of affiliative, maintenance and defensive behaviors in dogs (beerda et al. 1997; lindsay 2001; boissy et al. 2007; sontag and overall 2014). table 1. measures of physical health and cleanliness measure scale body condition score (bcs) 1=emaciated 2=thin 3=moderate 4=stout 5=obese body cleanliness (percent of the body covered in debris) 0=0% 1=1-25% 2=26-50% 3=51-75% 4= >76% nasal discharge present or absent ocular discharge present or absent tear staining present or absent sneezing present or absent coughing present or absent missing fur or poor coat present or absent wounds, sores or lesions present or absent lameness present or absent ninety seven dogs maintained at five commercial breeding kennels that were licensed by the usda in indiana, usa were scored on the physical and behavioral metrics compiled. during this initial scoring, two female observers approached the front of the dog's home kennel. the testers stood side by side 0.61m away from the kennel door, and turned their bodies so that their sides faced the door and dog to avoid direct eye contact that could have been aversive or threatening to the dog. the experimenter closest to the door bent at the waist and extended a hand toward the dog. using the ethogram, the dog’s immediate response to approach was recorded. the dog’s physical metrics were then recorded. following this scoring, the ethogram was revised to facilitate ease and accuracy of data collection obtained by live-observations in the field. categories were adjusted such that manifestations of the freeze, flight, and fight responses (lindsay 2001), were captured in a single category, fearful. behaviors that were never or only rarely observed during pilot testing were removed and behaviors that were observed and classified as "other" were added (table 3). page 3 developing the field instantaneous dog observation tool creative commons license 4.0 – non commercial – share alike – attribution bauer et al. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science category behavior description of behavior friendly/affiliative approach the dog moves toward the observer play bow the dog lowers the front half of its body while keeping the hindquarters elevated to indicate playfulness solicit attention the dog attempts to gain the observer’s attention (e.g. scrambling at the front of the pen) other maintenance eat the dog is consuming food rest the dog is laying down in a relaxed position groom the dog is cleaning itself other fidget/high arousal displacement behaviors the dog is demonstrating a normal behavior that is not appropriate to its current situation increased rr or harsh panting the dog is breathing at a more rapid rate than expected dilate pupils the dog's pupils are wider than would be expected given ambient light excessive vocalizations the dog vocalizes more than would normally be expected for the situation other fearful/avoidant move to back of enclosure/ outside the dog moves to the back of or leaves the kennel when approached climbing the walls the dog is attempting to scale the walls of its enclosure attempt to hide the dog moves to be unseen by the observer attempt to escape the dog is attempting to get away from the observer ambivalent (approach & avoid) the dog alternately moves toward and away from the observer displacement behaviors the dog is demonstrating a normal behavior that is out of context and reflects conflict trembling/ shaking the dog’s body is shaking crouched, slinking body posture the dog’s body is held low and close to the ground other fearful/shutdown freeze the dog holds its body completely still in the presence of the observer catatonic the dog is completely unresponsive in the presence of the observer trembling/ shaking the dog’s body is shaking other fear aggression offensive lunge the dog moves toward the observer quickly and aggressively vertical lip retraction the lips are pulled up to expose the front and canine teeth tail-up the tail is held tense, above hip level or vertical front of enclosure the dog is already at or moves to the front of the kennel when approached forward body posture the dog's weight and gaze are directed toward the observer hard body the dog’s entire body is tense and rigid other fear aggressiondefensive horizontal lip retraction the lips are pulled back to expose the molars withdrawal the dog moves away from the observer ambivalent (approach & avoid) the dog alternately moves toward and away from the observer backward body posture the dog's weight is directed away from the observer tail down the tail is held tense, below hip level other stereotypies circling the dog repeatedly moves in a circular pattern in the same direction pacing the dog walks back and forth in the kennel in a repeated pattern wall bouncing the dog jumps and pushes off of the walls of the kennel repeatedly other table 2. behavioral metrics used in assessment development page 4 www pet behaviour science org creative commons license 4.0 – non commercial – share alike – attribution 2017 | vol.3 | 1 15 category behavior description of behavior maintenance eat dog is consuming food rest dog is laying down in a relaxed position drink dog is consuming water affiliative approach dog moves toward the observer neutral wag the tail is held in a relaxed position at hip level and wagged solicit the dog attempts to gain the observer’s attention (e.g. scrambling at the front of the pen) play bow the dog lowers the front half of its body while keeping the hindquarters elevated to indicate playfulness lean near the dog leans on the door nearest to the observer fearful nonresponsive the dog sits frozen and does not respond to approach tail tuck the tail is tucked under the belly low wag the tail is held tense, below hip level, and wagged high wag the tail is held tense, above hip level or vertical, and wagged tremble the dog’s body is shaking crouch the dog’s body is held low and close to the ground hide the dog moves to be unseen by the observer growl the dog growls in response to approach bark the dog barks in response to approach hard body the dog’s entire body is tense and rigid hackles raised the fur on the back of the dogs neck is erect (piloerection) lip lift the lip is retracted either horizontally or vertically, exposing teeth lunge the dog moves toward the observer quickly and aggressively repetitive circling the dog repeatedly moves in a circular pattern in the same direction pacing the dog walks back and forth in the kennel in a repeated pattern wall bouncing the dog jumps and pushes off of the walls of the kennel repeatedly continuous vocalization the dog vocalizes without stopping calming behaviors paw lift one paw is lifted from the ground avert gaze the dog looks away from the observer yawn the dog yawns lip lick the dog licks its lips shake off the dog shakes its body after interaction with the observer positional front start in front of kennel the dog is already at the front of the kennel when approached move toward front of kennel the dog moves to the front of the kennel when approached positional back start in back of kennel the dog is at the back of the kennel when approached move toward back of kennel the dog moves to the back of the kennel when approached move from front to back repeatedly the dog moves from the front of the kennel to the back of the kennel and then returns to the front of the kennel repeatedly (or from the back to the front) other frantic the dog is highly excited play with waterer the dog manipulates the waterer when approached play with feeder the dog manipulates the feeder when approached table 3: categories of behavior and the behaviors included in each category pilot tested by expert raters study one: expert pilot testing and refinement methods two female experts in canine behavior and welfare science (one with a phd in applied animal ethology and the other an applied animal behaviorist with an ma in animal behavior) used the revised ethogramstyle form of the assessment to evaluate the response to approach of 38 dogs from two usda licensed commercial breeding facilities located in missouri, usa (facilities 1 and 2, table 4). facilities 1 2 3 4 5 total evaluator type (e expert; n novice) e e n n n number observed both with and without caretaker n/aa n/a 18 16 17 51 number observed without caretaker only n/a n/a 0 1 1 2 number observed with caretaker only n/a n/a 1 2 2 5 facility total 20 18 19 19 20 total by evaluator type • expert 38 • novice 58 total overall 96 a. n/a not applicable table 4: number of dogs observed at each facility. in all cases, the facility type was ‘breeder’ the approach test was performed in the manner described above. the raters indicated which behaviors in table 3 were performed in response to approach. these behavioral responses were then condensed into the behavioral categories. finally, the results were categorized into one of three rating codes: red, yellow, or green (ryg). responses classified as red included fearful behaviors (both offensive and defensive page 5 developing the field instantaneous dog observation tool creative commons license 4.0 – non commercial – share alike – attribution bauer et al. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science aggression indicators and shut down responses, such as freezing) and stereotypic behaviors. responses classified as green included affiliative behaviors such as approaching or soliciting attention from the rater, or neutral responses such as continued engagement in maintenance behaviors (e.g., eating, resting) despite the presence of the rater. dogs categorized as displaying a yellow response did not clearly respond to approach in a manner that could be classified as either green or red. irr, the ability of different raters utilizing the tool to reach the same conclusions (meagher, 2009) was evaluated. percent agreement was calculated for the physical metrics, individual behaviors, categories of behavior, and responses to approach classified as red, yellow or green based upon the recorded behaviors and notes made by the raters. percent agreement does not account for agreement due to chance, but can be an important baseline index of reliability (burn and weir 2011). in order to account for the presence of chance agreement, cohen’s kappa (landis and koch 1977) was used as a measure of irr for each of the documented behaviors and the behavioral categories and was calculated using spss statistics for windows, version 22.0 (ibm corp. released 2013. armonk, ny: ibm corp.). for the physical metrics recorded on an ordinal scale [body condition score (bcs) and body cleanliness] a weighted kappa statistic was calculated in sas, version 9.4 (sas institute inc., cary, nc) using proc freq with 0.0001 added to 0 values to facilitate the analysis. a weighted kappa statistic was also calculated for the ryg portion of the tool. kappa statistics <0.00 indicate poor agreement, 0.00-0.20 indicate slight agreement, 0.21-0.40 indicate fair agreement, 0.41-0.60 indicate moderate agreement, 0.61-0.80 indicate substantial agreement, and 0.81-1.00 indicate almost perfect agreement (landis and koch 1977). cohen’s kappa is influenced by homogeneity in responses and as the prevalence of a given response approaches either 0 or 100%, interpretation of the kappa estimate becomes more challenging. to aid in interpretation of the cohen’s kappa values, prevalence indices (pis) were calculated for the physical and behavioral components of the fido tool as a measure of homogeneity of response using the formula: pi = |g-o|/n where g indicates the number of agreed upon identifications of the behavior, o indicates the number of agreed upon instances where the behavior was not identified and n is the total number of observations (burn and weir 2011). the closer the pi is to 0, the more balanced the sample is in regard to the distribution of responses and the less influence prevalence has on the kappa estimate. the closer the pi is to 1, the more likely it is that sample homogeneity influenced the kappa estimate (burn and weir 2011). the scoring tool was further refined (table 5) so that only the behaviors regularly observed during the initial scoring sessions were included in the ryg categories on which novices were later trained. the dog’s position in the pen at the time of approach was removed at this stage of refinement since that information could generally be determined by the response to approach. testers were able to note the occurrence of any additional behaviors observed. category behavior description of behavior red fight the dog demonstrates offensively aggressive behavior (e.g. lunging at the observer) flight the dog moves to the back of or leaves the kennel when approached hard and forward body language the dog’s entire body is tense and rigid and oriented toward the observer frozen or catatonic the dog is completely still or unresponsive in the presence of the observer stereotypic behaviors the dog performs a pattern of behavior repeatedly yellow ambivalent body language the dog displays behavioral cues that may be interpreted in contradictory ways (e.g. growling or barking while tail wagging) ambivalent approach the dog alternately moves toward and away from the observer green friendly approach dog moves toward the observer solicits attention the dog attempts to gain the observer’s attention (e.g. scrambling at the front of the pen) neutral the dog is engaged in another activity (e.g. eating, resting, or play) and is undisturbed by the presence of the observer frantic/overstimulated the dog is highly excited table 5: behaviors included in the ryg evaluation page 6 www pet behaviour science org creative commons license 4.0 – non commercial – share alike – attribution 2017 | vol.3 | 1 15 results there was almost perfect agreement between the expert evaluators on the physical health metrics (table 6). measurement % agreement kappa (95% cia) pib bcs 100 1.00 0.95 c body cleanliness 94.74 0.48 (0.12, 1.00) 0.89 d nasal discharge 100 * 1.0 ocular discharge 100 1.00 0.95 tear staining 97.37 0.89 (0.69, 1.00) 0.71 sneezing 100 * 1.0 coughing 100 * 1.0 missing fur or poor coat 100 * 1.0 wounds, sores or lesions 100 * 1.0 lameness 100 * 1.0 table 6: percent agreement, kappa estimate of inter-rater reliability, and prevalence indices for the physical health attributed measured by expert raters. * indicates that agreement entirely or almost entirely in 1 category precluded calculation of a kappa statistic. a. ci confidence interval b. pi prevalence index / c. dogs were only rated as 3 or 4. d dogs were only rated as 0 or 1 percent agreement was 99.67% and kappa was 0.92 (95% ci: 0.77, 1.00). although bcs and body cleanliness were evaluated on an ordinal scale, only two levels of each variable were observed in this sample, allowing the use of cohen’s kappa and calculation of a pi for these variables. when the 1444 pairs of individual behaviors recorded in the initial ethogram-style assessment were evaluated, the overall percent agreement was 96.95% (n=1400), kappa was 0.86 (95% ci: 0.82, 0.90), and the pi was 0.77. table 7 provides the percent agreement, kappa, and pi for each of the behaviors in the ethogram-style assessment. a kappa value was unable to be calculated for 42.10% (16/38) of the individual behaviors because the raters either did not observe the behavior or it was very rarely identified. for an additional 23.68% of the behaviors (9/38), the raters were in perfect agreement. when the data were collapsed into behavioral categories, 304 pairs of evaluations were available for analysis. table 8 provides the percent agreement, kappa, and pi for each of the categories in this assessment. the overall percent agreement was 94.41% (n=287), kappa was 0.88 (95% ci: 0.82, 0.93), and the pi was 0.26. a kappa statistic was able to be calculated for all of the categories. the raters were in perfect agreement in two categories, maintenance and repetitive behaviors. the categories calm and fearful had low pis. thus, the substantial agreement indicated by their kappa values (0.74 and 0.73 respectively) could be considered reliable. when the data was further collapsed into ryg categories, an overall rating was made for each dog, giving a total of 38 pairs of evaluations. a green response to approach was identified by both raters for the majority of dogs (table 9). the overall percent agreement was 92.10% (n=35) with a kappa value of 0.87 (95% ci: 0.72, 1.000), indicating almost perfect agreement between the raters (landis & koch, 1977). a pi was not calculated because the weighted kappa was used to take into account all 3 outcome categories. category behavior % agreement kappa (95% cia) pib maintenance eat 100 1.00 0.89 rest 97.37 * 0.97 drink 100 1.00 0.89 affiliative approach 86.84 0.54(0.18-0.89) 0.66 neutral wag 84.21 0.66(0.43-0.90) 0.26 solicit 86.84 0.74(0.53-0.94) 0.13 play bow 100 1.00 0.68 lean near 100 * 1 fearful non-responsive 100 * 1 tail tuck 100 1.00 0.95 low wag 97.37 0.65(0.03-1.00) 0.92 high wag 100 1.00 0.95 tremble 100 1.00 0.95 crouch 100 * 1 hide 100 * 1 growl 94.74 0.48(-0.12-1.00) 0.89 bark 89.47 0.77(0.56-0.98) 0.32 hard body 97.37 0.84(0.54-1.00) 0.81 hackles raised 100 * 1 lip lift 100 * 1 lunge 100 * 1 page 7 developing the field instantaneous dog observation tool creative commons license 4.0 – non commercial – share alike – attribution bauer et al. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science category behavior % agreement kappa (95% cia) pib repetitive circling 100 1.00 0.84 pacing 100 * 1 wall bouncing 100 * 1 continuous vocalization 100 1.00 0.84 calming signal paw lift 100 1.00 0.95 avert gaze 97.37 * 0.97 yawn 97.37 0.65(0.01-1.00) 0.92 lip lick 86.84 0.65(0.38-0.93) 0.50 shake off 100 * 1 position start in front of kennel 89.47 0.54(0.15-0.93) 0.74 move toward front of kennel 100 * 1 start in back of kennel 97.37 0.84(0.54-1.00) 0.81 move toward back of kennel 94.74 0.72(0.35-1.00) 0.79 move from front to back repeatedly 94.74 * 0.95 other frantic 97.37 0.79(0.38-1.00) 0.87 play with waterer 100 * 1 play with feeder 100 * 1 table 7: percent agreement, kappa estimate of inter-rater reliability, and prevalence indices for the behaviors measured by expert raters in the ethogram-style assessment. * indicates that agreement entirely or almost entirely in 1 category precluded calculation of a kappa statistic for this behavior. a. ci confidence interval b. pi prevalence index study two: novice pilot testing methods to assess the ability of the ryg scoring tool to be reliably used by novices in canine behavior, two female undergraduate student raters with limited formal training in canine behavior and welfare evaluated the response to approach of 58 dogs from three usda licensed commercial breeding facilities located in indiana, usa (facilities 3, 4, and 5). % agreement kappa (95% cia) pib maintenance 100 1.00 0.79 affiliative 94.74 0.64 (0.17-1.00) 0.84 fearful 86.84 0.73 (0.52-0.94) 0.18 repetitive 100 1.00 0.74 calm 89.47 0.74 (0.51-0.98) 0.42 positional front 89.47 0.54 (0.15-0.93) 0.74 positional back 97.37 0.91 (0.73-1.00) 0.66 other 86.84 0.79 (0.38-1.00) 0.87 overall 94.41 0.88 (0.82-0.93) 0.26 table 8: categorical agreement when assessed by expert raters. a. ci confidence interval b. pi prevalence index rater 1 rater 2 n. red (%) 5 (13.16) 4 (10.53) n. yellow (%) 4 (10.53) 7 (18.42) n. green (%) 29 (76.31) 27 (71.05) total 38 38 table 9: summary of dogs with response to approach rated as red, yellow or green by expert raters prior to evaluating the dogs, these raters were trained on how to perform the approach and identify behaviors falling into the ryg categories. as with the expert raters, the rating pair approached the dog with body angled perpendicular to the kennel door. the experimenter closest to the door bent at the waist and extended a hand toward the dog. each dog’s immediate response to approach was recorded as red, yellow, or green. on the first approach, only the raters were present. the raters approached the front of the kennel, noted the selected dog’s response to approach and then evaluated the metrics of physical well-being. after each selected dog at the facility was evaluated, the raters waited for 30 minutes, then repeated the test with a caretaker present in order to gauge whether dogs’ responses differed with a familiar person. table 4 details the number of dogs observed at each facility and, for the novice raters, those observed without a caretaker present, with a caretaker present, and both with and without a caretaker present page 8 www pet behaviour science org creative commons license 4.0 – non commercial – share alike – attribution 2017 | vol.3 | 1 15 percent agreement was calculated for the physical metrics and responses to approach classified as red, yellow or green. as described in the expert pilot testing section, cohen's kappa was calculated for physical metrics recorded as present or absent as a measure of irr. a weighted kappa was utilized for the physical metrics recorded on an ordinal scale and the ryg portion of the tool. additionally, the percentage of dogs with response to approach in each category was calculated for approaches conducted with and without the caretaker present at the individual facility level and for the overall sample. where the raters agreed upon the dogs' responses to approach a wilcoxon signed rank test was performed to determine if the presence of a caretaker made a statistically significant difference in the responses to approach using spss statistics for windows, version 22.0 (ibm corp. released 2013. armonk, ny: ibm corp.). measurement % agreement kappa (95% cia) pib bcs 82.98 -0.04 (-0.11, 0.03) 0.83 body cleanliness 67.39 0.37 (0.15, 0.60) n/a nasal discharge 100 * 1.0 ocular discharge 90.12 * 0.90 tear staining 68.08 0.20 (-0.00, 0.40) 0.55 sneezing 100 * 1.0 coughing 100 * 1.0 missing fur or poor coat 100 * 1.0 wounds, sores or lesions 100 * 1.0 lameness 100 * 1.0 table 10: percent agreement, kappa estimate of inter-rater reliability, and prevalence indices for the physical health attributed measured by novice raters. * indicates that agreement entirely or almost entirely in 1 category precluded calculation of a kappa statistic. a prevalence index was not calculated for body cleanliness as this was assessed on an ordinal scale with a weighted kappa. a. ci confidence interval b.pi-prevalence index results when the binary indicators of physical well-being were evaluated in aggregate, percent agreement between the novice raters was 96.29% and kappa was 0.26 (95% ci: 0.02, 0.50), fair agreement. the kappa value was strongly influenced by a homogeneous sample as indicated by the overall pi of 0.95. table 10 provides the percent agreement, kappa, and pi for each of the indicators of physical well-being. the novice raters were in agreement as to the type of response to approach of 81.13% of the 53 dogs observed when a caretaker was not present (n=43). when a caretaker was present, the novice raters were in agreement as to the type of response to approach of 85.71% of the 56 dogs observed (n=48). table 11 displays each rater’s classification of responses to approach with and without a caretaker present. no caretaker caretaker rater 3 rater 4 rater 3 rater 4 n. red (%) 15 (28.30) 9 (16.98) 8 (14.28) 6 (10.71) n. yellow (%) 4 (7.55) 8 (15.09) 12 (21.43) 15 (26.79) n. green (%) 34 (64.15) 36 (67.92) 36 (64.29) 35 (62.50) total 53 53 56 56 table 11: summary of dogs with response to approach rated as red, yellow or green with and without caretaker presence by novice raters interrater reliability table 12 displays percent agreement and kappa values for the behavioral portion of the fido tool with and without a caretaker present. because irr reflects an assessment of the tool, rather than the subjects of observation, it is important to evaluate the raters’ responses in aggregate and thus the facility level values are not reported. when all of the observations were considered regardless of caretaker presence, the raters agreed 83.49% of the time (91/109). the weighted kappa value was 0.74 (95% ci: 0.63, 0.85), indicating substantial agreement (landis and koch 1977). a pi was not calculated because the weighted kappa was used to take into account all 3 outcome categories and at this time a pi or equivalent has not been developed for the weighted kappa (burn and weir 2011). page 9 developing the field instantaneous dog observation tool creative commons license 4.0 – non commercial – share alike – attribution bauer et al. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science without caretaker present with caretaker present overall % agreement k (95% cia) % agreement k (95% ci) % agreement k (95% ci) 81.13 0.72 (0.56 0.87) 85.71 0.76 (0.61 0.92) 83.49 0.74 (0.63 0.85) table 12: percent agreement and weighted kappa for the ryg assessment by caretaker presence when performed by novice evaluators. a. ci confidence interval effect of caretaker presence fifty-one dogs were observed both with and without a caretaker present. the raters agreed on the response to approach of 37 dogs (72.55%). table 13 displays the percentage of responses to approach classified in each of the ryg categories when the raters agreed upon the dog’s response to approach both with and without a caretaker. there was a high proportion of green responses to approach both without a caretaker present (81.08%, n=30) and with a caretaker present (78.38%, n=29). the percentage of yellow responses increased when a caretaker was present (16.21%, n=6 versus 2.70%, n=1). the percentage of red responses decreased when a caretaker was present (5.40%, n=2 versus 16.22%, n=6). upon analysis of the agreed-upon paired samples from all breeding facilities using a wilcoxon signed rank test the presence of a caretaker did not produce a statistically significant change in response to approach (t=-0.69; n=37; p=0.490). discussion the first goal of this study was to develop a field-ready tool for use by laypersons in behavior to quickly and accurately gauge the immediate health and welfare status of kenneled dogs. we also aimed to pilot-test the tool for irr, and evaluate the effect of caretaker presence in the context of commercial breeding facilities. importantly, the fido tool developed was intended to provide a useful gauge of key aspects of dog welfare status at the time of observation in a field evaluation, as is often needed for routine site inspections or investigation of animal welfare complaints. under such circumstances, it is important that inspectors or investigators have reliable, standardized, animal-based metrics for use to facilitate consistent, objective evaluations. the fido tool is not intended to be used as a comprehensive assessment of the long-term welfare status of dogs. however, fido may provide an important screening tool that permits users to determine if and where more detailed dog welfare assessments are needed. red % (n) yellow % (n) green % (n) total n wilcoxon t (p) facility 3 without caretaker present 33.33 (5) 0.00 (0) 66.67 (10) 15 -0.63 (0.529) with caretaker present 13.33 (2) 33.33 (5) 53.33 (8) 15 facility 4 without caretaker present 0.00 (0) 10.00 (1) 90.00 (9) 10 -0.49 (0.627) with caretaker present 0.00 (0) 0.00 (0) 100.0 0 10 10 facility 5 without caretaker present 8.33 (1) 0.00 (0) 91.67 (11) 12 -1.51 (0.132) with caretaker present 0.00 (0) 8.33 (1) 91.67 (11) 12 all facilities without caretaker present 16.22 (6) 2.70 (1) 81.08 (30) 37 -0.69 (0.490) with caretaker present 5.40 (2) 16.22 (6) 78.38 (29) 37 table 13: percentage of types of agreed upon response to approach expressed by dogs at commercial breeding facilities based on caretaker presence. while behavioral and welfare assessment tools have been developed with a focus on dogs living with families and housed in animal shelters (kiddie and collins, 2014; belshaw et al. 2015; barnard et al. 2016) and tools for the evaluation of personality traits in working dogs have also been developed (sinn et al. 2010) this tool is unique. first, it was developed for those who need to quickly and accurately evaluate the physical and behavioral status of breeding dogs in the field without the use of video-recordings or other methodologies and resources that normally permit detailed analyses of long term behavioral patterns (beerda et al. 1999; hepper and wells 2000). second, the tool was developed to be particularly useful and applicable for evaluating dogs housed in commercial page 10 www pet behaviour science org creative commons license 4.0 – non commercial – share alike – attribution 2017 | vol.3 | 1 15 breeding facilities. further, it is the only tool to our knowledge reported to have been tested on-site with this particular population of dogs. the experiences of these dogs are likely to be very different from those housed in shelters as they may have limited exposure to environments and people outside of the breeding kennel. this has implications for those kennels that rehome their dogs, as dogs rehomed from commercial breeding operations have been reported to be more likely to exhibit behaviors associated with fear than companion dogs raised in other settings (mcmillan et al. 2011). thus, the dogs’ responses to approach as a measure of fear (and a surrogate measure of socialization) were evaluated. percent agreement, irr, and pis were calculated as part of the development of the tool. as part of pilot testing the ryg behavioral assessment portion of the tool, irr was determined and the effect of caretaker presence was evaluated. third, the tool not only allows an evaluator to indirectly assess the dog’s socialization to strangers, it also may provide insight into the quality of the relationship with the dog’s caretakers when scoring is done with and without their presence. the expert evaluators had near perfect agreement on the physical metrics of the dogs evaluated. the novices likewise had high levels of agreement here, suggesting that the tool has high irr on these components. additionally, the experts had very high levels of percent agreement and kappa values that indicated almost perfect agreement when the data were coded as individual behaviors. however, the high or low prevalence of certain individual behaviors prohibited the calculation of cohen’s kappa for many of the behaviors. when investigating a construct, such as fear of strangers, a rating system such as the ryg system used in this study may be a better choice than coding individual behaviors (kubinyi et al. 2015). in a comparison of predictive validity between behavioral and trait (or construct) ratings for success in a working dog training program, only slight differences in the predictive validity between methods were detected (wilsson and sinn 2012). similarly, in this study, despite moving from behavioral coding to construct ratings, almost perfect inter-rater reliability was maintained as indicated by kappa values greater than 0.8 (landis and koch 1977). when the ryg rating system was used by the novice raters, percent agreement was also high while the kappa statistic indicated substantial agreement (between 0.61 and 0.80) (landis and koch 1977). although this tool is intended for use by raters without expertise in canine behavior, our findings suggest that additional training for raters in both behavior and the use of the tool may further improve irr. as the majority of dogs evaluated by the novice raters demonstrated a green response to approach, it is possible that homogeneity of responses influenced the kappa statistic. further evaluation of the tool in kennels of different types and management styles may elicit more variation in responses, which may help to confirm the strong reliability of the tool. the prominence of green responses to approach may be challenging analytically, but it also provides insight into the behavior of commercial breeding dogs at their home facilities. the high proportion of green responses to approach contrasts with previous reports of high levels of fear in rehomed breeding dogs (mcmillan et al. 2011). the discrepancies may be due to the fact that the approach tests were performed on-site at the dogs' home pens and they were therefore unlikely to have experienced the stressors that cumulatively might result in the global fear observed in other studies focusing on owner reports or observations made after leaving the commercial breeding environment. the owners of the breeding facilities chose to participate in the study, and thus the facilities included here may not be representative of all commercial breeding facilities. additionally, this test has not yet been validated against long term metrics of dog well-being and therefore might only reflect the immediate welfare status. further evaluation is needed to determine if this is indeed the case. the findings of this study may also potentially provide insight into facility management and the quality (and perhaps frequency) of interactions dogs are having with people they regularly encounter. overall, the presence of a caretaker in this study resulted in a decrease in the proportion of responses to approach classified as red. although no statistically significant difference was detected across facilities when a caretaker was present versus absent, different patterns in behavioral responses to approach by strangers with page 11 developing the field instantaneous dog observation tool creative commons license 4.0 – non commercial – share alike – attribution bauer et al. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science and without caretaker presence emerged at the individual facilities. if indeed response to approach is a useful surrogate measure for socialization, such changes may indicate the effectiveness of different management and socialization programs on dogs’ responses to both caretakers and strangers. for example, in facilities where the proportion of red responses to approach decreased when the primary caretaker was present, it is likely that social buffering of stress induced by the presence of strangers occurred. such patterns have been reported in other species (boissy et al. 1998; hennessy et al. 2000; ruis et al. 2001; hennessy et al. 2009; croney 2014). this would suggest that the dogs were well socialized to their caretakers, and likely had positive interactions with them. however, keeping in mind breed variation relative to selection for or against affinity for strangers, having a high percentage of dogs showing red responses to strangers approaching in a nonthreatening manner may indicate a need for improved socialization to people. likewise, in facilities where the proportion of green responses to approach decreased when the primary caretaker was present, poor or insufficient interactions with caretakers may be indicated. thus, there is potential for this tool to help identify facilities and individual dogs within facilities that would benefit from improved socialization efforts. there is also potential to monitor and potentially improve the quality of dog-caretaker interactions. overall, the assessment tool evaluated in this study appears to be a useful means of quickly gaining some insight into the immediate welfare status of kenneled dogs. there has been debate about the validity of behavioral assessment tools in predicting aggressive behavior in dogs (kis et al. 2014; patronek and bradley 2016). although the tool studied in this paper focused on capturing fearful behaviors and fear may manifest in aggression (lindsay 2001; sonntag and overall 2014), additional testing of its validity is needed before anything might be concluded about its predictive power. instead, our initial testing focused on reliability given the need for consistency of behavior identification in those tasked with on-site evaluations of dogs. as further evidence of the need for such consistency, in a study of 60 observers, the groups of dog owners, veterinarians, dog trainers, and non-dog owners did not differ in ability to identify behavior, but there were differences between individuals within and between all categories (tami and gallagher 2009). comparing agreement between expert and novice raters is a necessary next step in the evaluation of reliability and a crucial step in determining that the novice raters are interpreting responses to approach in the same manner as experts. in addition to being reliable, robust welfare assessment tools must be valid (meager 2009). this pilot study’s major limitation is that the instantaneous metrics of well-being used have not yet been validated against those reflective of long-term welfare. however, this tool is not intended to replace the in-depth, detailed or longer term assessments of dog welfare which are needed to fully understand dogs’ states relative to their attempts to cope with their environments (broom 1991). nor is it intended to stand alone as a predictor of a dog’s ability to be an acceptable companion. to accomplish these goals, the validity of the metrics used would need to be investigated in various ways. for example, comparing the results of this tool to validated tools as has been done with other species (powell et al. 2016), or to physiologic measures such as cortisol levels would be helpful in confirming that the response to approach by a stranger is correlated with overall welfare status. further, scoring dogs outside of their home pens and in response to different stimuli would lend additional insight into both the behavioral wellbeing of dogs and the validity of the tool. these limitations notwithstanding, the assessment does appear to offer good reliability and is potentially useful in identifying individual dogs potentially in need of interventions who appear at least initially to be fearful upon approach. given this, while this study focused on the commercial dog breeding industry, the tool has the potential to be of benefit to people working with dogs housed in other types of kennels in other types of industries, including animal shelters and research facilities. future studies will focus on evaluating the effects of additional training for novices, applying the fido tool to more diverse groups of dogs in different types of facilities, and testing the validity of the tool through comparisons between novice and expert raters and through comparisons with other methods of evaluating welfare in dogs housed in kennels. page 12 www pet behaviour science org creative commons license 4.0 – non commercial – share alike – attribution 2017 | vol.3 | 1 15 conclusion this study aimed to develop and assess the reliability of the fido tool, a non-invasive, instantaneous field ready evaluation of the welfare status of dogs housed in commercial breeding facilities. this tool is intended to be useful to kennel inspectors, caretakers and others who require the ability to quickly gauge dogs’ welfare states. overall, the tool provides a useful means to assess individual dogs, identify those having difficulty coping with their environment, and inform recommendations to improve dog welfare in kennel settings. it may therefore be helpful to commercial breeding facilities aiming to promote the physical, emotional, and behavioral well-being of their breeding dogs and puppies. further, although refinement of training methods and further testing in more diverse environments are needed, this tool has the potential to be useful in other types of facilities where dogs are kenneled. acknowledgements funding for this project was provided by the world pet association and the pet food institute. mary jordan's participation was supported by the merial veterinary scholars summer research program. monica colon's participation was supported by the purdue university summer research opportunities program. the authors would like to thank dr. judith stella for her critical review of the manuscript. 7. references barnard, s., pedernera, c., candeloro, l., ferri, n., velarde a., and dalla villa, p. 2016. development of a new 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10.1016/j.jveb.2016.10.002. page 15 developing the field instantaneous dog observation tool creative commons license 4.0 – non commercial – share alike – attribution bauer et al. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science the oxytocin receptor gene, an integral piece of the evolution of canis familaris from canis lupus. the oxytocin receptor gene, an integral piece of the evolution of canis familaris from canis lupus jessica lee oliva1, yen t wong2, jean-loup rault3, belinda appleton2, alan lill4 highlights this study revealed genomic differences between the domestic dog and the wolf. these differences were close to the oxytocin receptor gene and implicate this gene in the dog’s evolution from the wolf. pet behaviour science | 2016, vol.2, 1-15 jessica lee oliva1, yen t wong2, jean-loup rault3, belinda appleton2, alan lill4 1. school of biological sciences, monash university 2. life and environmental sciences, deakin university, geelong 3.animal welfare science centre, university of melbourne. 4. department of ecology, environment and evolution, school of life sciences, la trobe university, melbourne paper research email: jss_oliva@yahoo.com.au australia. keywords: dog, gene, object, oxytocin, wolf although some dogs are naturally better than other dogs at solving such tasks, this study did not reveal any genomic differences between good versus poor performing domestic dogs. while further genetic studies are warranted, this suggests that other nongenetic influences may effect object choice task performance, such as the early life experiences of the dog. 1. introduction the neuropeptide, oxytocin, is commonly known for its role in mammalian bonding (lim and young 2006). several studies have also found an association between plasma oxytocin (handlin et al 2001; odendaal and meintjes 2003), serum oxytocin (miller et al. 2009), or urinary oxytocin (nagsawa et al. 2009), and human-dog bonding. however, caution should be applied when interpreting these findings for two reasons. firstly, while odendaal and meintjes used established chromatographic methods to measure oxytocin, handlin et al. and miller et al. employed enzyme immunoassay methods without prior sample extraction, which has recently been shown to be page 1creative common license 4.0 – non commercial – share alike – attribution 2016 | vol. 2 | 115 abstract previous research in canids has revealed both group (dog versus wolf) and individual differences in object choice task (oct) performance. these differences might be explained by variation in the oxytocin receptor (oxtr) gene, as intranasally administered oxytocin has recently been shown to improve performance on this task by domestic dogs. this study looked at microsatellites at various distances from the oxtr gene to determine whether there was an association between this gene and: i) species (dog/wolf) and ii) good versus bad oct performers. ten primer sets were designed to amplify 10 microsatellites that were identified at various distances from the canine oxtr gene. we used 94 (52 males, 42 females) blood samples from shelter dogs, 75 (33 males, 42 females) saliva samples from pet dogs and 12 (6 males, 6 females) captive wolf saliva samples to carry out our analyses. significant species differences were found in the two markers closest to the oxtr gene, suggesting that this gene may have played an important part in the domestic dogs’ evolution from the wolf. however, no significant, meaningful differences were found in microsatellites between good versus bad oct performers, which suggests that other factors, such as different training and socialisation experiences, probably impacted task performance. http://www.petbehaviourscience.org/ necessary in obtaining valid results (christensen et al. 2014; szeto et al. 2011). similarly, nagasawa and colleagues validated their methodology for measuring oxytocin in dogs’ (but not human) urine using plasma samples assayed with no mention of extraction (mitsui et al. 2011). however we can assume extraction did not take place due to the high values they obtained, typical from unextracted samples, out of the normal biological range (in humans) measured using validated methods (christensen et al.; mccullough et al. 2013). secondly, and more importantly, peripheral measures of oxytocin are not necessarily a reflection of central function. indeed, studies have demonstrated that oxytocin, as well as its structurally similar neuropeptide, arginine vasopressin, do not readily cross the blood-brain barrier (robinson 1983; veening et al. 2010; vorherr et al. 1968) and can be independently released in the brain and in the body (amico et al. 1990; engelmann et al. 2000; ludwig and leng 2006; robinson and jones 1982). an alternative method to better evaluate the effects of central oxytocin function is to administer oxytocin intranasally. although their mechanisms of action are poorly understood (leng and lugwig 2016), neuropeptides administered in this way are believed to gain access to the brain, as increased levels can be found in brain dialysates (neumann et al. 2013) and cerebrospinal fluid (csf) (born et al. 2002; dal monte et al. 2014; rault 2016; striepens et al. 2013) following administration. therefore, rather than measuring the peptide itself, behaviours can be measured after peptide administration. this method has been exploited in many human studies and oxytocin is now receiving increasing attention for its role in human social cognition, as demonstrated by increasing gaze to the eye region of human faces (guastella et al. 2008), improving perception of human movements (keri and benedek 2009), enhancing facial processing, emotion recognition, memory and encoding of facial stimuli (see review, guastella and macleod 2012), and increasing trust (kosfeld et al. 2005). it is important to note, however, that the effects of oxytocin have not always been pro-social; it has caused decreases in sociality under certain circumstances, for example, in experiments that involve out-groups, competition and/or rejection sensitive participants (see review, bartz et al. 2011). in dogs, intranasally administering oxytocin has been shown to increase positive expectations (kis et al. 2015), decrease friendliness in response to a threatening person (hernádi et al. 2015), increase affiliation towards owners and conspecifics (romero et al. 2014), increase play behaviours towards conspecifics (romero et al. 2015) and increase mutual gaze with their owners (nagasawa et al. 2015). our recent study involving intranasal oxytocin administration in domestic dogs (canis familiaris) found that it also increased the ability of dogs to use human communicative gestures to find food in an object choice task (oct) (oliva et al. 2015). dogs were tested on this task on two separate days, 515 days apart, once after oxytocin administration and once after saline. order of treatments were predetermined according to group allocation, oxy-sal or sal-oxy. the task involved a human experimenter using momentary distal pointing cues to indicate the location of a hidden food reward in one of two opaque containers to the right and to the left of her. results showed that the dogs that received oxytocin in their first testing session (oxy-sal dogs) outperformed dogs that received saline, and this performance was maintained up to 15 days later, in the absence of further oxytocin administration, where they performed equally as well as dogs receiving oxytocin in their second session (sal-oxy dogs) (oliva et al. 2015). we suggest that changes in the canine oxytocinergic system may have been integral to the evolution of the domestic dog from the wolf (canis lupus), the phylogenetically closest species to the domestic dog (wang et al. 2013). evidence supporting this theory comes from studies which have shown that whilst domesticated dogs perform well on octs employing relatively difficult momentary distal pointing cues (miklósi et al. 2005; soproni et al. 2002; virányi et al. 2008), hand-reared wolves do not (miklósi et al. 2003; virányi et al.), at least without extensive training (virányi et al.). moreover, extensive socialization with humans does not further enhance domestic dogs’ ability to use momentary distal pointing cues at four months old. this is evidenced by the fact that, with hand-reared domestic dogs who were separated from their mothers a few days after birth and intensively hand-reared by humans, and pet dogs who remained with their mothers for several weeks after birth before being adopted out to a human family, performed equally well on octs employing this cue. the same page 2 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.2 | 1-15 intensive socialization did not help hand-reared wolves of the same age perform the same task as they demonstrated an ability no better than chance. however, these wolves did demonstrate an ability to learn this cue over time and at age eleven months were able to perform at above chance level, comparable to naïve dogs of the same age (virányi et al.). further evidence for the important influence of learning in wolves comes from a study by udell et al. (2008b) which demonstrated that when adult pet dogs and adult hand-reared wolves were both tested in an outdoor enclosure (an environment familiar to wolves but unfamiliar to dogs) using momentary distal pointing cues given by a familiar experimenter, wolves significantly outperformed dogs. however, pet dogs that were tested indoors, even by an unfamiliar experimenter, performed as well as the wolves tested outdoors. this was not true for shelter dogs tested indoors. the reason that the wolves in udell et al.’s study were readily able to use this cue, while the wolves in miklósi (2003) et al.’s study who were tested in a similar environment were not, may be due to the fact that, as pointed out by hare et al. (2010, p.e6): “the wolves that udell and colleagues tested probably had received previous training” and udell and wynne (2010) agree with this suggestion in a later publication. but this does not explain why the dogs in this study performed so poorly. ried (2009) has suggested that the olfactory, auditory and visual contact with conspecifics in this experimental set-up may have been unfairly distracting to the dogs who were likely to be unfamiliar with the other dogs in the experiment, compared with the wolves who were used to their conspecific companions. this would suggest that an important factor in performing accurately on an oct is a familiar or natural environment. indeed, results from dogs tested in an unnatural environment (kirchhofer et al. 2012; udell et al. 2008b; udell et al. 2010b) (behind a fence, outside or in a shelter) suggest that their ability can be compromised by environmental factors that probably impact on the animals’ level of stress. nagasawa et al. (2015)’s study has provided further evidence supporting the claim that the oxytocinergic system may have been shaped through the process of domestication in a way which allowed for dogs to both communicate and bond with humans. this was evidenced by the fact that mutual dog-owner gaze (which increased measures of oxytocin in the owner’s urine) increased in female dogs following intranasal oxytocin administration and was not achieved with hand-raised wolves, who only rarely demonstrated mutual gazing with their owners. geneticists are continuing to investigate the association between human social cognition and the oxytocin receptor (oxtr) gene. for example, single nucleotide polymorphisms (snps) have been associated with attachment style (chen et al. 2011; denes 2015), and autism (jacob et al. 2007; wu et al. 2005), as well as less extreme disorders of social functioning such as the presentation of callous-unemotional traits and conduct problems (dadds et al. 2014). associations have also been found between oxtr gene snps and mothers’ sensitivity to their toddlers (bakermans-kranenburg and van ijzendoorn 2008; riem et al. 2011), empathy and stress (rodrigues et al. 2009), optimism, personal mastery and self-esteem (saphire-bernstein et al. 2011), positive affect and non-verbal intelligence (lucht et al. 2009), and sociality (tost et al. 2010). in addition, associations have been found between oxtr gene snps and amounts of oxytocinergic-rich connections in the hypothalamus and connections between the hypothalamus and both the amygdala and the dorsal anterior cingulate cortex (tost et al. 2010). whilst there is strong evidence to support variants of this gene functioning differently in humans, there is little evidence to date supporting differing effects of variants of this gene in dogs. it is reasonable to assume that this is likely, however, given the vast array of homologous human-dog phenotypes that have been associated with analogous genes in the two species (see reviews, parker et al. 2010; sutter and ostrander 2004). allelic variability of the canine oxtr gene is poorly known. one study investigated snps within the oxtr gene in dogs and found associations with proximityseeking and friendliness in two different dog breeds (kis et al. 2014). however, the snp associated with friendliness was associated with opposing phenotypes in the two different breeds. similar findings have been found with oxtr gene snps which are associated with autism affecting people of different ethnicities page 3 the oxytocin receptor gene in wolf-dog evolution creative common license 4.0 – non commercial – share alike – attribution oliva et al. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science differently (jacob et al. 2007; wu et al. 2005). an alternative approach to increase the probability of finding genomic differences, especially in a relative small sample of dogs of different breeds, is to look at microsatellites close to the gene of interest. microsatellites are short tandem repeats that can occur within both the coding and non-coding (un-translated) regions of the genome and have been used extensively to demonstrate association between a genomic location and a trait (see review, montaldo and meza-herrera 1998). despite the fact that dogs generally perform well on octs when tested in a natural environment, many studies report a wide range of individual variability in performance (agnetta et al. 2000; hare et al. 2002; miklósi et al. 1998; udell et al. 2008a; udell et al. 2008b, udell et al. 2010b; virányi et al. 2008; wobber et al. 2009) and this may reflect differences in the oxtr gene. in dogs, the oxtr gene is 2.41 million base pairs long and located on chromosome 20 at position 9.36 million base pairs within the genome (ncbi 2012). we used microsatellites located at different distances from the oxtr gene (refer to table 1) to investigate genetic association with oct performance phenotypes. given the species differences demonstrated in several studies (hare et al. 2002; miklósi et al. 2003; udell et al. 2008b; virányi et al. 2008), the objectives of the current study were to search for genotypic differences near the oxtr gene between: i) good and poor oct performers and ii) domestic dogs and wolves. 2. methods study animals owners of 75 pet dogs (33 males, 42 females) volunteered their dogs for a study investigating the effect of intranasal oxytocin on dogs’ performance on an oct. dogs were required to be more than 12 months old, healthy, and not pregnant or lactating. no restriction was put on breed, as this was too difficult to control using pet and shelter dogs whose breeds were often mixed and not confirmable. recruitment took place through poster advertisements at university campuses and on university e-newsletters and social media websites. to increase the effective sample size of domestic dogs for genetic analysis, an additional 94 blood samples (52 from males, 42 from females) were obtained from the animal aid animal shelter, coldstream, victoria, australia. blood, rather than saliva, was used as a matter of convenience as 0.5-1ml blood samples were routinely collected from shelter dogs to test for heart-worm, and it was easy to take an extra 2-2.5 ml for the purposes of our study. twelve samples of wolf saliva were supplied by wolf park, battle ground, indiana, usa. all wolves were a mix of the subspecies: arctos, occidetalis and/or nubilus, and had been human-reared from 10-14 days of age and whilst were living as a pack in a large outdoor enclosure, still interacted with humans on a daily basis. the study was approved by the monash university school of biological sciences animal ethics committee (bsci/2013/07). materials intranasal treatments containing either oxytocin dissolved in saline, or containing saline only were stored in frozen tubes labelled ‘a’ or ‘b’. both the experimenter and the pet dog’s owner were ‘blind’ as to which tubes contained which treatments. twenty-four international units of oxytocin (auspep, melbourne, au) dissolved in a 0.1 ml saline solution was administered to the nostrils of the dogs via a mucosal atomizer device connected to a 1ml syringe, with a half dose (0.05 ml) in each nostril. dogs received each treatment on a separate testing day in a predetermined, pseudo-randomized and counterbalanced order. oragene animal swabs (canada) were used to collect deoxyribonucleic acid (dna) samples in saliva of the pet dogs involved in the study, as well as from the wolves. approximately 2-2.5 ml blood samples were drawn from the cephalic or jugular vein of the shelter dogs. prior to collection, the area was clipped with surgical blades and cleaned with methylated spirits. ten tandem repeats were visually identified close to the dog oxtr gene (ncbi 2012) and primers were designed using the online program, primer 3 (untergasser et al. 2012; see table 1). ‘lamb puffs’ were used as dog treats in the oct and were concealed using four identical, opaque spaniel bowls. pen and paper were used to score performance (correct/incorrect choice of concealed food reward) on the oct. page 4 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.2 | 1-15 procedure when the pet dog arrived at the testing location for their first testing session, he/she was administered an intranasal treatment of saline or oxytocin, depending on which group he/she has been pseudo-randomly assigned to: those that received oxytocin in their first session and saline in their second session (oxy-sal) or vice versa (sal-oxy). when the dog arrived for their second session, they received the other intranasal treatment and there was a 5-15 days washout period between sessions. after the intranasal administration a swab was rubbed against the inside of the animals’ cheeks for approximately 30 seconds and stored in a fridge at the testing location (this usually occurred in session 1) before being transported to the laboratory for genetic analysis. the dog was required to wait for 45 minutes after receiving the nasal spray before the testing began. during this time the dog was free to roam the testing room, interact with its owner and/or the experimenter, or wander outside. after the 45 minute waiting period lapsed, the oct commenced, which involved a) pre-training (before each 10 trial block of the oct proper): four correct trials in a row where the dog was shown a food treat being dropped into one of two dog bowls, either side of the experimenter (delivered using the ipsilateral hand from a kneeling position, 75 cm from the target) (b) 20 trials (block 1 and block 3) where the dog was given a momentary distal pointing cue (a 1-2 second point delivered using the ipsilateral index finger from the same kneeling position) to indicate the location of the hidden food reward located in one of the two dog bowls and (c) 20 trials (block 2 and block 4) using a gaze cue (a 1-2 second gaze shift delivered from the same kneeling position, keeping the head straight). five control trials per block, where no cue was given, were also delivered to ensure the dogs were not relying on scent to solve the task. the task usually lasted between 45 and 60 minutes and was carried out by the experimenter with the help of the owner, who called the dog back to the starting position between trials and restrained it until the experimenter said the release word “ok”. the experimenter recorded the score for each trial as either correct (if the dog approached the bowl with the hidden food treat) or incorrect (if the dog approached the empty bowl or the experimenter) before starting the next trial. trials were also considered incorrect if the dog made no choice but was page 5 the oxytocin receptor gene in wolf-dog evolution creative common license 4.0 – non commercial – share alike – attribution oliva et al. table 1. microsatellites and their distances from oxtr gene within the canine genome. the oxtr gene is located at 9.36 million base pairs within the genome (ncbi 2012). https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science deemed motivated to complete the task by participating when two of the easier pre-training cues were then given as a test of motivation. for the purposes of the current study, only the momentary distal pointing data were analysed because dogs were not able to use the gazing cue above chance levels (oliva et al. 2015). this data is the same as that which was published in oliva et al. sample preparation and polymerase chain reaction ninety-four genomic dna samples were extracted from shelter dogs’ blood using axyprep blood genomic dna miniprep kit (axygen, usa). eightyseven saliva samples were obtained from pet dogs and wolves using oragene animal/saliva kits (oragene, canada). polymerase chain reaction (pcr) was performed in a total of 12.5 l volume reactions in a 96 well pcr plate (interpath services, australia). each well contained a 6.25 l aliquot of genomic dna and a 6.25 l pcr reagent mix. the pcr reagent mix contained distilled water, 2.5 mm mgcl2 , 1mm deoxynucleotide triphosphates (dntps), 1x pcr buffer, 1u of taq polymerase, 0.28 µm fluorescent-labelled m13(-21) primer (fam, vic, pet or ned) (promega, usa), 0.072 µm forward primer and 0.28 µm reverse primer (macrogen, korea) per reaction. for all the forward primers the 5’ end was modified with a m13(21) universal sequence tag (5’-tgtaaaacggccagt3’) to enable the incorporation of the universal fluorescent labelled m13(-21) primer for detection on abi3730 capillary instrument (macrogen, korea) (schuelke, 2000). all pcrs were performed with a veriti 96 well fast thermal cycler (applied biosystem, australia). the thermal cycler was programmed to 1 cycle of 5 min at 94˚c as initial hot start, then 30 sec at 94˚c, annealing step of 30 sec at 55-65˚c and extension step of 40 sec at 72˚c. this was followed by a repeat of the cycle above 30 times and then by 8 extra cycles to ensure the oligo dye attached to the maximum amount of fragments. then followed denaturation at 94˚c for 30 sec, annealing at 53˚c for 45 sec and extension at 72˚c for 45 sec. finally, 1 cycle of 10 min at 72˚c was run for final extension and held at 14˚c. electrophoresis of amplified products after amplification, a 2 l aliquot of the amplified pcr product was combined with 2 l of loading buffer (0.4% (w/v) bromo-phenol blue, 0.5m edta and 6 ml of glycerol) and analyzed directly on 1% (w/v) agarose le (benchmark scientific, australia) gel in 1 x tae buffer (50 mm tris acetate, 1 mm edta). two l of hyper ladder i (bioline, australia) was used as a size marker to compare the molecular weight of the amplified products. gels were run at 100 volts for 25 mins and the gel images were documented by molecular imager chemi doc xrs4 imaging system (syngene, uk). then, 4 l of each of four different microsatellite amplicons were pooled for the same animal. these pooled samples were combined into a master 96 well plate and sequenced by macrogen (korea). data analysis deoxyribonucleic acid fragment analysis was carried out using nucleic acid analysis software (toonan and hughes 2001). this software allowed fast analysis of the multiplexed microsatellite markers. contingency tables were used to compare the case and control canine with the risk and wild type genotypes. this allowed assessment of the degree of association of performance and species traits. in line with recommendations by campbell (2007), contingency tables with expected frequencies > 1 were analysed with an n-1 χ2, and contingency tables with expected frequencies < 1 were analysed with a fisher-irwin test by irwin’s rule. these analyses were carried out using spss version 22.0 ( spss ibm, new york, usa, 2013), following methods from weaver (2013). given the observed effect of intranasal oxytocin on performance in the oct which lasted across the sessions for dogs that received oxytocin in session 1 (oliva et al. 2015), we separated good performing versus poor performing dogs in the following three ways so as not to confound the results. firstly, we looked at only the group of dogs that received saline in their first testing session (sal-oxy dogs) and used performance data from that session only. secondly, because there was no difference between the performance level of the two groups of dogs in session 2 (with oxy-sal dogs having maintained their enhanced performance from session 1, and sal-oxy dogs having their performance enhanced by oxytocin in session 2), we used “oxytocin-induced performance” data (i.e. session 2 data) from all dogs in the study. we considered good performers as dogs that scored 18/20 page 6 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.2 | 1-15 correct points, and poor performers as those that scored ≤ 12/20 correct points. dogs scoring between 1317/20 were excluded from the analysis. finally, we wanted to see if some dogs were more susceptible than other dogs to the oxytocin treatment and so analysed difference scores in performance between the two sessions for sal-oxy dogs only. we considered high oxytocin responders to be dogs that improved their performance by 3-7 points between sessions, and poor responders as those whose performance remained the same, or declined, between sessions. dogs scoring a difference of 1-2 points between sessions were excluded from the analysis. 3. results primers 9-10 were not analysed because they did not anneal correctly to the target template. as such, only results pertaining to primers 1-8 are shown below. tables 2-4 show the microsatellite markers identified for the analyses. within the sal-oxy group of dogs, there were 13 poor performers, five good performers, and 15 mid-range performers that were excluded from the analysis. there was no significant association between session 1 performance and any of the primers (table 2). in session 2, there were 10 poor performers (5 oxy-sal dogs and 5 sal-oxy dogs), 28 good performers (19 oxysal dogs and 9 sal-oxy dogs), and 24 mid-range performers (8 oxy-sal dogs and 16 sal-oxy dogs) that were excluded from the analysis. there was no significant association between session 2 performance and any of the primers (table 3). when evaluating difference scores between sessions for the sal-oxy group of dogs, there were 10 poor oxytocin responders, 13 high responders, and eight mid-range responders that were excluded from the analysis. there was no significant association between oxytocin response and any of the primers (table 4). finally, we tested whether there were any differences between the oxtr gene of the domestic dog and that of the wolf. table 5 shows the microsatellite markers identified for the analysis. tables 6-7 show the contingency tables for the significant analyses. there was a significant association between species and primer 5, p = 0.038 (table 5). odds ratio for the allele (risk/wild-type) = 12.00, indicating that a canine with the risk allele is twelve times more likely to be a dog than a wolf, than if it has a wild-type allele. page 7 the oxytocin receptor gene in wolf-dog evolution creative common license 4.0 – non commercial – share alike – attribution oliva et al. table 2. microsatellite markers and session 1 performance association analysis by contingency n-1 χ2 or fisher-irwin test by irwin’s rule. table 3. microsatellite markers and session 2 performance association analysis by contingency n-1 χ2 or fisher-irwin test by irwin’s rule. table 4. microsatellite markers and difference in performance between sessions association analysis by contingency n-1 χ2 or fisher-irwin test by irwin’s rule. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science there was also a significant association between species and primer 6, p = 0.026 (table 5). odds ratio for allele (risk/wild-type) = 11.61, indicating that a canine with the risk allele is almost twelve times more likely to be a dog than a wolf, than if it has a wild-type allele. 4. discussion the study revealed significant species differences between dogs and wolves, using 2 microsatellite primers close to the oxtr gene. considering that the domestication of the dog occurred approximately 32,000 years ago (wang et al. 2013), the identified allelic differences between dog and wolf are likely to reflect an old mutation. in an old mutation under strong selection, we expect linkage disequilibrium to decay rapidly as we move away from the causative gene. therefore, we would expect to see a strong association between genetic variation and species near the oxtr gene and no association when we move along the chromosome away from the gene. this study found that two markers close to the oxtr gene, primers 5 and 6, were different in dogs and wolves. of all the primers, these are the two closest to the oxtr gene at positions 9.11 million and 9.66 million base pairs, respectively, supporting the above-hypothesized strong selection. for primer 5, the wolves did not demonstrate a common allele, probably due to the very small number of wolf samples that amplified (refer to table 6). by examining the raw data, we see that all of these samples expressed a different microsatellite size, one of which was the risk allele. for primer 6, the risk allele was not present in any of the 6 wolf samples that amplified and could be analysed (refer to table 7). due to the small sample sizes used in the current study, it is difficult to draw any firm conclusions, however, given that we are seeing significance in markers closest to the gene and an absence of significance in markers further away from it, these findings certainly warrant further research to be carried out with a larger sample of wolf dna. another limitation of our study was that the blood samples from shelter dogs, and the saliva samples from pet dogs and wolves, were analysed in two separate batches with no blood-saliva samples that had come from the same canine which we could use to standardize the samples. as such, while unexpected, there is no way of knowing if there were biases in the samples and therefore, future studies should be mindful to collect additional samples from a subgroup of animals on which to carry out this standardization. future studies could also utilize other markers in this region to map the association more accurately e.g. snps. the current study failed to detect genomic differences in goodversus poor -performing dogs on an oct. this may be because we were not focusing closely enough on the oxtr gene itself. due to the novelty of the study and the unknown variation within the canine oxtr gene, rather than looking within the gene, we looked at microsatellites close to the gene to increase the chance of finding any association. however, it is possible that we have missed a genetic difference within the gene itself that differentiates good from poor oct performing domestic dogs. dog breed, which was not controlled in the current study, may also affect behavioural phenotypes as observed by kis et al. (2014) with respect to friendliness, and jakovcevic et al. (2010) with respect to gaze towards the human face. lastly, the transmembrane enzyme, cd38, is attracting research interest, as variations of this gene have been page 8 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2016 | vol.2 | 1-15 table 5. microsatellite markers and species association analysis by contingency n-1 χ2 or fisher-irwin test by irwin’s rule. *p <.05 table 6. contingency table for primer 5 and primer 6. linked with oxytocin secretion and associated social behaviours (see review, macdonald 2012). hence, future studies should also investigate this gene, in addition to the oxtr gene, when examining phenotypes believed to be governed by the oxytocin system. alternatively, the lack of significant disparities between good and poor performances may be due to the fact these differences are not genetic in nature. for example, there may have been differences in the individual dogs’ learning and training. indeed, dogs seem to possess a degree of cognitive flexibility in their use of human cues, for example, learning not to follow human pointing in an oct when it is no longer reinforcing, and can even learn to avoid a pointed at object if a non pointed at object suddenly becomes the reinforcer (elgier et al. 2009). furthermore, if dogs have learned to associate the hidden food reward in an oct with a physical (non-human) cue this may then hinder their ability to successfully use a human pointing cue (elgier et al. 2012). prior training and learning was not investigated in the current study but may be responsible for a dogs’ level of performance. socialisation history is also an important factor which may explain individual differences in oct performances. for example, early stimulation of the oxytocinergic system, for example, via hormonal imprinting, has been shown to have implications for function in later life (see review, csaba 2000). as csaba explains, a critical window exists in the first few days after birth in which this hormonal imprinting can occur. the process is defined by the provocation of a hormone receptor-to-be by a circulating hormone. if this process does not occur, the receptor does not mature and is unable to bind with the hormone in a suitable quantity. for most receptor cells, this inability is life-long and passed down to daughter cells (except in the brain where most cells do not differentiate) and in some cases, even to offspring of the animal. in his review, csaba (2008) surmises that this is a result of a change in heritable dna methylation; however, the exact mechanism behind hormonal imprinting remains unknown. faulty imprinting can occur as a result of a lack of the appropriate hormone which reduces receptor density (csaba 2008). csaba (2000; 2008) supports the notion of imprinting as a “memory-like process” (csaba 2000, p. 409) in which short repeated bouts of exposure result in greater imprinting than one single, long exposure. faulty imprinting likely explains why adult rats demonstrate significantly less oxytocin or vasopressin receptors in the central nucleus of the amygdalae and the bed nucleus of the stria terminalis after receiving low levels of maternal licking and grooming as pups, compared to those who received high levels of maternal licking and grooming (francis et al. 2002). tanaka et al. (2010) also demonstrated similar findings whereby isolation-reared rat pups have fewer immunoreactive vasopressin cells (males) in the dorsal, medial parvocellular part of the paraventricular nucleus of the hypothalamus, than socially-reared pups, and fewer oxytocin cells (females) in the ventral, medial parvocellular part of the paraventricular nucleus of the hypothalamus. isolation-reared males were also more cautious than socially-reared ones in an elevated plus-maze and neither male nor female rat pups displayed signs of familiarity during a social recognition test. faulty imprinting may also explain findings from a study of nursery-reared rhesus monkeys that displayed significantly less oxytocin in their cerebrospinal fluid and less affiliative and more aggressive and abnormal repetitive behaviours compared to mother-reared rhesus monkeys. moreover, a retrospective study found that women with a history of childhood abuse, particularly repeated and emotional abuse, displayed lower levels of oxytocin in their cerebrospinal fluid in adulthood (heim et al. 2009). in addition to this critical period for hormonal imprinting, there is another critical period for socialisation in birds and mammals, in which they willingly accept other animals, including humans, into their environment (scott 1962). if socialisation with humans does not occur within a dogs’ critical period, 310 weeks of age (scott), a fear of humans may override their cognitive capacity to use humans’ social cues in a beneficial way. this lead udell et al. (2010a) to the ‘two stage hypothesis’ which states that sensitivity to human cues requires i) interactions with, and acceptance of, humans during the sensitive period of the canid’s social development, followed by ii) learning, through classical page 9 the oxytocin receptor gene in wolf-dog evolution creative common license 4.0 – non commercial – share alike – attribution oliva et al. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science and operant conditioning, to pair human gestures with the acquisition of something favourable. this has been supported by a study demonstrating better oct performance by pet dogs versus purpose-bred research dogs that have been socialised to humans at a young age, but who do not cohabit with them and have relatively less contact with them than pet dogs have with their owners (lazarowski and dorman 2015). all the dogs in our study were currently-owned pets whose backgrounds were not investigated, however, while some had come from responsible breeders, others were shelter ‘rescues’, and others came from pet shops renowned in australia for selling puppies obtained from puppy farms (rspca 2010). dogs born into poor environments, such as puppy farms, may not have experienced adequate imprinting of oxytocin to its receptors, or adequate socialisation within the critical period due to deprivation of positive social experiences, and may have comprised the poor performers in our study for this reason. this possibility requires further research from future studies that should take dogs’ socialisation backgrounds into account. 5. conclusions by measuring microsatellites close to the oxtr gene in samples of domestic dogs and wolves, this study provides evidence that mutations in this gene may have played a part in the domestic dog’s evolution from the wolf. the study did not, however, produce evidence for the same mutations 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between dogs and humans. nature communications 4. doi: 10.1038/ncomms2814 weaver b. 2013. assumptions/restrictions for chisquare tests on contingency tables. retrieved march 27, 2015 wobber v., hare b., koler-matznick j., wrangham r., tomasello m. 2009. breed differences in domestic dogs' (canis familiaris) comprehension of human communicative signals. interaction studies 10(2): 206224. wu s., jia m., ruan y., liu j., guo y., shuang m., gong x., zhang y., yang x., zhang d. 2005. positive association of the oxytocin receptor gene (oxtr) with autism in the chinese han population. biological psychiatry 58(1): 74-77. doi: 10.1016/j.biopsych.2005.03.013 page 15 the oxytocin receptor gene in wolf-dog evolution creative common license 4.0 – non commercial – share alike – attribution oliva et al. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science influence of dog presence on the tolerance and evaluation of aversive stimulation comparing trainers’ reports of clicker use to the use of clickers in applied research studies: methodological differences may explain conflicting results lynna c. feng*, tiffani j. howell, pauleen c. bennett pet behaviour science | 2017, vol.3, 1 – 18 doi: 10.21071/pbs.v0i3.5786 lynna c. feng*, tiffani j. howell, pauleen c. bennett school of psychology and public health, la trobe university. p.o. box 199, bendigo, victoria 3552 paper research * corresponding author: l.feng@latrobe.edu.au australia. keywords: clicker training; dog learning; dog trainer perceptions; dog training; learning theory highlights • clicker training is both a training technique and a philosophical approach to dog training • dog trainers recommend using a clicker-type signal when training novel behaviours but not when rewarding known behaviours • a mismatch exists between applied and experimental criteria for sufficient signal conditioning • empirical studies and individual perceptions disagree on the relative efficacy of clickers versus other signals page 1creative common license 4.0 – non commercial – share alike – attribution 2017 | vol. 3 | 1 18 abstract clicker training refers to an animal training technique, derived from laboratory-based studies of animal learning and behaviour, in which a reward-predicting signal is delivered immediately following performance of a desired behaviour, and is subsequently followed by a reward. while clicker training is popular amongst dog training practitioners, scientific evaluation in applied settings has been largely unsuccessful in replicating the benefits of reward-predicting signals seen in laboratory animal studies. here we present an analysis of dog trainers’ advice and perceptions, conducted to better understand clicker training as it occurs in the dog training industry. twentyfive sources (13 interviews with dog trainers, 5 websites, and 7 books) were analysed using a deductive content analysis procedure. we found that, for many sources, “clicker training” referred not only to the technique, but also to a philosophy of training that emphasises positive reinforcement and the deliberate application of learning theory principles. many sources reported that clicker training was fun, for both dog and handler, but that it could be frustrating for handlers to learn and sometimes cumbersome to juggle the extra equipment. in addition, while most sources recommended clicker training particularly when training new behaviours, many stated that it was no longer needed once the dog had learned the desired behaviour. when comparing industry recommendations to methods used in applied studies, different criteria were used for predictor signal conditioning. inadequate conditioning of the predictor signal in empirical evaluations could partly explain the lack of learning benefits in applied studies. while future research is needed to verify the practitioner beliefs in a wider population, these results provide an in-depth description of what clicker training is, at least for the sources analysed, and a potential starting point for understanding methodological factors that could contribute to previous studies’ failure to demonstrate the benefits purported to exist by industry practitioners. http://www.petbehaviourscience.org/ 1. introduction clicker training is an animal training technique that employs a clicker (hand-held device that makes a clicking sound when pressed) or other signal to predict the subsequent presentation of a reward (i.e. something the animal wants). originally introduced to the dog training industry by karen pryor in her book called don’t shoot the dog! (historical account by gillaspy et al. 2014; pryor 1999), clicker training is currently extremely popular. a google keyword search in august 2016 for the term “clicker training” yielded approximately 1.35 million hits, and karen pryor clicker training reported over 1400 clicker trainers attending dedicated clickerexpo seminars in 2016 (clayton 2016). somewhat surprisingly, then, a review by feng et al. (2016) reported that “clicker training” itself is poorly defined and motivations underlying the use of clicker training have yet to be systematically investigated. fundamentally, clicker training is derived from laboratory-based studies on operant conditioning (skinner 1938) and the search for mechanisms capable of reducing the adverse effects of delayed reinforcement on learning (e.g. grice 1948). laboratory studies suggest that animals learn most quickly when they receive immediate consequences after performing a desired behaviour, but learn more slowly (as reviewed by lattal 2010) or fail to learn the task at all (browne et al. 2013) when the consequences are delayed even by a very short interval. the use of a predictor signal, such as the clicker, that predicts the presentation of a subsequent reward, can mitigate the detrimental effects of delayed primary reinforcement (i.e. a time delay between the desired behaviour and reinforcing consequence) in laboratory settings (lattal 1984). in applied training contexts, delayed reinforcement is often inevitable (e.g. reinforcing behaviour that is performed at a distance). immediate predictor signals, such as the click of a clicker, have therefore been proposed to mitigate the learning deficit that would normally follow such delays (as reviewed by feng et al. 2016). empirical investigations of the efficacy of clicker training in animal learning typically compare the effect of an auditory reward-predicting signal (such as a clicker) followed by a primary reinforcer with the effect of a primary reinforcer alone (control group). such studies have been conducted in the following companion animal species: horses (mccall and burgin 2002; williams et al. 2004), dwarf goats (langbein et al. 2007), and dogs (blandina n.d.; chiandetti et al. 2016; smith and davis 2008). as reviewed by feng et al. (2016), the findings are surprisingly inconclusive. while goats appeared to learn a shape discrimination task more quickly in a predictor signal treatment group as compared to a control group (langbein et al. 2007), none of the other studies found the predictor signal treatment condition to be more effective than the treatment used with the control group. in fact, an unpublished thesis reported that dogs in a food-only (control) treatment group reached a higher learning criteria than those in the predictor signal treatment groups (clicker+food and verbal “next”+food) (blandina n.d.). numerous differences across and within the study methodologies, and in comparison with prior laboratory research, could contribute to these seemingly paradoxical findings. to the best of our knowledge, however, behavioural scientists have not yet experimentally evaluated the methodological variations present in these studies. these include: how to introduce the predictor signal, the type of predictor signal used, the task being trained, and the presence and location of a human trainer (as reviewed by feng et al. 2016). considering the lack of consistency in methodologies used in empirical studies, the primary aim of this study was to rigorously describe and critically evaluate clicker training methodologies used by or recommended by animal trainers. it was expected that this would provide a better understanding of how clicker training is implemented in applied settings. we discuss this information in the context of available scientific literature and propose additional avenues of research into how methodological differences might influence the efficacy of clicker training protocols. 2. methods advice and perceptions regarding “clicker training” were collected from three sources: interviews with selfidentified dog trainers, bestselling books on clicker training, and clicker training websites. the interviews page 2 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2017 | vol.3 | 1 18 conducted for this project were approved by the la trobe university human research ethics committee (approval number: s15-274). all sources were subjected to a content analysis which is described in more detail below. interviews with dog trainers a convenience sample of volunteers (n=13; 12 women and one man) were interviewed for this study. all were self-identified dog trainers, fluent in english and ranging in age from 24 – 73 years (m = 41 years; sd = 17.59). recruitment was conducted at a dedicated clicker training conference (reno, nevada), online through social media, and via word of mouth until data saturation was reached and no new information was obtained (as suggested by morse 1995). each participant completed a brief demographic survey and scheduled an interview at his/her convenience. semistructured interviews were conducted, in which overall topics included: the participant’s experience with clicker training, their beliefs regarding when it is appropriate, and whether it is an effective learning tool. the interview schedule was open ended and participants were encouraged to elaborate on topics they raised. interviews were held in person, via telephone, or online by web-conference between january-march 2016. all interviews were audio recorded and transcribed for analysis. interview participants lived in the united states (n=7) and australia (n=6). all participants self-identified as dog trainers in response to the question “on a scale from 1 (completely disagree) to 5 (completely agree), to what degree do you identify with the statement ‘i am a dog trainer’” (completely agree n=12, somewhat agree n=1). although the interview recruitment mentioned clicker training, the advertisement was purposely designed to recruit both trainers who used clickers (hereafter referred to as clicker trainers) and those who did not, in hopes of capturing a range of perspectives. the resulting interviewees were skewed towards identifying as clicker trainers, but the sample included a number of trainers who did not use clickers. in response to the question “on a scale from 1 (completely disagree) to 5 (completely agree), to what degree do you identify with the statement ‘i am a clicker trainer’” participants responded as follows: completely agree (n=6), somewhat agree (n=2), neither agree nor disagree (n=2), somewhat disagree (n=2), completely disagree (n=1). for the purpose of having the thoughts and perceptions of trainers who both used and did not use clickers represented in the qualitative analysis, rather than trying to draw comparisons between these individuals and any other sample, this sample was deemed sufficient. books about clicker training the primary objective when choosing books for this review was to examine the most readily available sources of information accessed by everyday dog owners. it was felt that this would be the most objective method of determining predominant advice and practices. as such, a non-personalised google books search was conducted with the search terms ‘clicker training dog’ in november 2015, with the top five results (excluding advertisements) selected for review. the books selected were as follows, in the order they appeared in the google search: • book1: spector, m. (1999). clicker training for obedience. sunshine books. • book2: pryor, k. (2002). getting started: clicker training for dogs. sunshine books. • book3: fisher, g. t. (2009). the thinking dog: crossover to clicker training. dogwise publishing. • book4: ray, m. (2008). click & train your dog: using clicker training to transform your common canine into a superdog. tfh publications, inc. • book5: meagher, j. m. (2014). the wonder of clicker training: the complete guide to a nonviolent, positive, compassionate, & effective way of dog training with clickers. createspace publishing. of these books, three appeared to be guides for dog owners interested in clicker training their own dogs (book2, book4, book5), while the remaining two appeared to be written for individuals, such as trainers, more interested in the nuances of clicker training and why it should be used (book1, book3). page 3 clicker training: research versus practice creative common license 4.0 – non commercial – share alike – attribution feng et al. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science websites about clicker training as with choosing books to review, the goal in selecting websites on clicker training was to choose the most readily available sources, believed most likely to reflect industry recommendations received by dog owners. hence, the top five results (excluding advertisements) from a google.com query for the term ‘clicker training’ were selected. the non-personalised search was conducted in march 2016 and the google.com database algorithm yielded the following results: • web1: karen pryor clicker training: the leader in positive reinforcement training http://www.clickertraining.com/ • web2: clicker training wikipedia, the free encyclopedia https://en.wikipedia.org/wiki/clicker_training • web3: clicker training lessons http://www.clickerlessons.com/whatis.htm • web4: clicker training center: learn to clicker train your dog. http://www.clickertrain.com • web5: clickersolutions training articles http://www.clickersolutions.com/articles similar to the books selected, there appeared to be websites written for both dog owners interested in training their own dogs (web2, web4) and those with more in-depth discussions on clicker training practices and controversies, perhaps targeted more towards professional or semi-professional trainers (web1, web3, web5). preliminary analysis a cursory glance at the list of books and websites selected for analysis indicates that choosing sources based on search engine results, rather than purposefully selecting a variety of source from different backgrounds, may have resulted in three potential problems. first, all sources focused on basic pet dog training, rather than on training for other activities, such as working or performance roles, where clicker training is very popular (e.g. gerritsen et al. 2013; vegh and bertilsson 2010). second, there appeared to be a bias towards sources stemming from karen pryor clicker training. third, a website entry on clicker training from the online collaborative encyclopaedia wikipedia was the second highest website search result. to deal with the first potential problem, it was decided to include two additional books, purposely selected to represent the use of clickers in training dogs for working roles and competitive dog sports. the topranking books in an amazon.com search of “dog sport clicker training” and “working dog clicker training” sorted by popularity were chosen in december 2015. the books selected appeared to be targeted towards individuals who wanted to become agility dog trainers and service dog trainers, respectively: • book6: bertilsson, e. & johnson vegh, e. (2010). agility right from the start: the ultimate training guide to america's fastestgrowing dog sport. sunshine books. • book7: shaw, r. (2015). service dog training – guide dogs, hearing dogs, therapy dogs, working dogs, puppies, pet therapy, emotional support, disabled, clicker training, registration, certification – all covered. clovelly publishing. while we were cautious about including too much information from karen pryor clicker training, due to the company’s dominant role in the industry, the repeated appearance of associated sources was to be expected. as such, it was decided to proceed as planned. similarly, while we are aware of the known risks of relying on information from wikipedia (as discussed by hilles 2014), it was noted that the opinions presented in the wikipedia article were not in conflict with those voiced on the other websites. consequently, we chose to include this website. since it was ranked second in the google.com query we conducted, it is likely a leading source of advice on clicker training. main analysis to analyse the sources, the deductive qualitative content analysis process described by elo and kyngäs (2008) was followed. the purpose was to describe the phenomenon of clicker training within the framework of pre-defined questions. an attempt was made to page 4 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2017 | vol.3 | 1 18 http://www.clickertraining.com/ http://www.clickersolutions.com/articles http://www.clickertrain.com/ http://www.clickerlessons.com/whatis.htm https://en.wikipedia.org/wiki/clicker_training address the following questions: 1) what is clicker training? 2) why do people use clicker training? and 3) what methods are generally considered ‘best practice’ in relation to clicker training? to address these questions, an unconstrained categorisation matrix was developed, where original categories were created based on the overarching questions of interest and a preliminary reading of all sources. following initial transcription and coding, all coded data from the interviews (int1-int13), books (book1-book7), and websites (web1-web5) were pooled in the hope of generating a broad sense of ideas and perceptions present in the industry. because this potentially results in some confusion about the source of specific information, we noted where ideas were consistent across source types and, in the analysis reported below, provide the source of sample quotes where appropriate. during the coding process, sub-categories were created following elo and kyngäs’ (2008) inductive content analysis approach of creating additional categories that describe the phenomena outside of the pre-determined categories. this allowed for flexibility in expanding on topics based on the information provided by the source materials, while maintaining a focus on the questions of interest. finally, relevant empirical studies were analysed for the categories that were generated in order to draw methodological comparisons. based on the analyses, we present a synopsis of the results and, where applicable, discuss the findings within the context of available empirical studies. 3. results and discussion what is clicker training? the first question addressed what came to mind when people thought about the term “clicker training”. when defining clicker training, two general aspects were described (figure 1). sources defined clicker training in terms of either a training philosophy or a technique. some even recognised the term as having a dualfaceted nature. web5 stated: “clicker training is both a training technique and a training philosophy”. clicker training as a philosophy as a philosophy, sources such as web3 stated that engaging in clicker training meant “using no physical compulsion or corrections whatsoever…using almost entirely positive reinforcement”, with an emphasis on applying the “[scientific] principles of learning” (web5). in addition, sources mentioned the development of a partnership and emphasised a positive experience for both the dog and handler. according to int9, “it's a partnership that forms…i feel more in tune with the dog”. these philosophical aspects of the definition of clicker training were thought to contrast with other popular dog training methods. for example, when comparing clicker training to other methods she had previously page 5 clicker training: research versus practice creative common license 4.0 – non commercial – share alike – attribution feng et al. figure 1. coded categories with regards to the question: “what do people understand by the term ‘clicker training’?” https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science used, int9 described the following: “it's more… teamwork rather than a dictatorship”. clicker training as a technique in terms of the technique of clicker training, most sources described it as a technique employing the use of a clicker to identify desirable behaviours and predict subsequent food rewards. a typical statement was that clicker training is a training method in which the handler “[uses] a ‘clicker,’ a mechanical device that makes a short, distinct ‘click’ sound which tells the animal exactly when they're doing the right thing” (web1). however, some sources indicated that the sound did not necessarily need to be from a clicker device. instead it could be, as int4 said, “some [other] kind of marker signal…not always a clicker” such as a spoken word or a whistle, and would still be considered under the umbrella term “clicker training”. what appeared to be emphasised was the idea that the signal, whether it was a clicker or otherwise, had to be a form of communication to the dog. multiple interviewees suggested that, particularly when a dog was first being introduced to the signal, “the click means the food is coming” (int8), but with practice, the signal begins “[communicating] to the dog that that was what i wanted you to do” (int12). dog perceptions in clicker training next, we investigated how people thought dogs perceived the signal. in 2016, the authors reviewed three potential theoretical hypotheses for mechanisms underlying clicker training: reinforcing, bridging, and marking (feng et al. 2016). the theoretical review found that clicker-type reward-predicting signals most likely function as a secondary reinforcer, but could have bridging and marking properties as well, where the primary function of the clicker-type predictor signal depended on the context in which it was applied. when sources were analysed to investigate industry perceptions of clicker functioning, only a few believed that the clicker functioned as a reinforcer. int13 stated that “the dog should feel the same way about the ‘yes’ that it feels about the food.” web5 suggested that, with sufficient pairing, “the click means the same thing to the dog that the presentation of food would.” however, not all sources took this position. int6 stated “i don't think it's a reward.” book5 suggested something similar to the bridging mechanism instead: “[the] clicker serves…to bridge the gap between the time the dog performs the desired behavior and the time they receive the treat.” on the other hand, many sources reported that the more important function of the signal was to “[tell] the animal exactly when they're doing the right thing,” as stated by web1. similar to the marking hypothesis, multiple sources described the clicker as “an event marker” (web5). however, unlike the marking function described in the scientific literature, where the event marker merely emphasises a point in time without providing reinforcing or punitive feedback, multiple sources believed that the clicker signal communicates to the dog not just “the precise time the dog did the behaviour” (int10) but that “they're doing the right thing” (web1) and specifically communicates “what [behaviour] they're being rewarded for” (int8). for example, instead of using the signal to broadly mean that the dog had done something correct, int12 said that “giving the clicker sound is like a photograph of the correct behaviour.” based on these results, it appears that there is not a single, widely accepted consensus on what “clicker training” is. nor do people agree on what it means (or should mean) to the animal receiving the click. this could potentially explain why it has proved so difficult to demonstrate in applied settings that the clicker is an effective tool. perhaps those researchers conducting the studies are using the tool in a different way to how it is used in applied settings. scientific evaluation of clickers in both dogs (smith and davis 2008) and horses (williams et al. 2004) used protocols that tested the clicker for its power as a secondary reinforcer. williams et al. (2004) found no effect of the clicker as a secondary reinforcer in horses. likewise, smith and davis (2008) also found no difference in rate of task acquisition between clicker and non-clicker groups. however, they did find that dogs rewarded with the click sound alone after performing a behaviour continued to do so for longer than those who received no feedback after performing the behaviour. this suggests that the click was acting as a secondary reinforcer in this context. given that sources tended to focus more on the communicative effects of the clicker than its power as a reinforcer, it is possible that the context in which page 6 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2017 | vol.3 | 1 18 studies were investigating clicker training differed from how practitioners use it in an applied setting. this could explain the discrepancy between practitioner experiences and empirical results. regardless, clicker training continues to grow in popularity. the following section reports factors that sources suggest motivate people to use clicker training and those that discourage people from wanting to use clicker training. why do people clicker train? the second question we aimed to answer was “why do people clicker train?” to investigate this question, we coded sources based on arguments for and against clicker training relating to the dog, the handler, and the handler-dog relationship (see figure 2). arguments for clicker training consistent with previously published reports of industry perceptions (smith and davis 2008), sources suggested that clicker training helps dogs learn faster. however, sources additionally reported numerous other benefits of clicker training for the dog (figure 2). one unexpected benefit that was reported across multiple sources was that clicker training “encourages [the dog] to think for himself” (book4) and become a “fully active, thinking participant” (web1). as int9 explained, this characteristic of clicker training particularly sets it apart from more traditional training methods where “command compliance” is valued as opposed to nurturing active learning and problem solving skills. furthermore, sources reported that, with clicker training, dogs appeared eager to learn; book6 described “a happy and confident dog” and stated that “clicker training will turn your dog into a ‘learning junkie,’ a dog who is eager to offer behaviors and to experiment to get you to reward.” to the best of our knowledge, these page 7 clicker training: research versus practice creative common license 4.0 – non commercial – share alike – attribution feng et al. figure 2. coded categories with regards to the question: “why do people use clicker training?” https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science additional benefits of clicker training have not yet been scientifically evaluated, nor have empirical studies been published using dogs or horses who had prior experience with clicker training. perhaps benefits such as active learning and problem solving develop over time, resulting in more efficient learning after a dog becomes experienced with clicker training. int9 suggested that “to actually really be a solid clicker training dog would…probably take two months.” if so, this could explain why empirical studies, which normally test animals with no previous clicker training experience after a small number of click-treat pairings (20 or fewer in blandina n.d.; smith and davis 2008; williams et al. 2004; wood 2007), fail to replicate the enhanced rates of learning reported by clicker training practitioners. for the most part, sources reported handler benefits that matched those of the dogs. clicker training was thought to help people learn to train more effectively. sources attributed this benefit to the focus on precision and timing of reinforcements, and the emphasis on the handler attending to specific criteria in the dog’s behaviour during training sessions. in addition, int4 suggested that clicker training gave handlers an opportunity to “see the little lightbulb” when the dog understood a new task and int5 said that trainers “[start] to better understand how a dog actually learns.” with regards to the handler-dog relationship, multiple sources stated that clicker training “reinforces the bond between dog and trainer” (book5) and improves communication between dog and handler by providing “a dialogue between trainer and dog” (book7). arguments against clicker training although the sources analysed in this study were heavily biased towards those who support the use of clicker training (all books and websites were on the use of clicker training, and 8 out of the 13 interviewees identified as clicker trainers) there were a number of recurring arguments presented against clicker training. as with the benefits, sources reported costs relating to the dog, handler, and the dog-handler relationship. all reported costs are included in figure 2. most commonly, sources stated that clicker training can be time consuming and frustrating for the handler to learn. this came in the form of statements such as: “clicker training is too difficult for beginners” (web5), “can…be frustrating” (int6), and “unless you [have] got an actual [person] to learn from, it is actually quite hard” (int5). many people also noted that handlers were often unable or unwilling to carry and handle a clicker device at all times. int10 described a training situation as follows: “i can't handle it because [i have] a lead in one hand, i've got treats here, the dog's pulling this way and that way and i can't handle it… the clicker is in the way.” however, these individuals generally chose to use a verbal signal, such as “yes” or “good,” in place of the mechanical click, rather than using no signal at all: “i found it was easier for me to always have my voice on me than to make sure i always have a clicker on me” (int13). in addition to frustration with learning the mechanical skills, book3 suggested that, when first learning to use clicker training, handlers might feel “loss of control,” which could be related to the coordination required or the less “command-oriented” training philosophy. in light of the reported difficulty and frustration experienced by handlers first learning to clicker train, these consequences should be carefully considered. with regards to the handler-dog relationship, int7 voiced concern that using clicker training techniques was “a lot of bribery” and the handler was “putting [themselves] into the position of a servant.” web5 agreed that some “dogs will work only when food or the clicker is present.” however, web5 clarified that this issue happens when “food is misused” and to instead “reward, don’t bribe…the food or toy should be produced only after the dog has performed the behavior.” these concerns suggest that clicker training can have negative consequences when used in certain ways. in addition, the concerns presented here have been collected from a sample that predominantly supports the use of clicker training. as such, these reported costs of clicker training are likely not fully representative of the beliefs and experiences of those who do not support the use of clicker training and future research is warranted to better understand these concerns. advice and recommendations many of the arguments against clicker training related to handlers successfully learning to implement clicker training techniques. it is not surprising, then, that sources provided a myriad of rules and suggestions for how clicker training should be implemented (see page 8 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2017 | vol.3 | 1 18 figure 3). the third question addressed was: “what methods are generally considered ‘best practice’ in relation to clicker training?” overall, there were four broad categories of advice offered: 1) how the signal should be introduced, 2) what signals should be used, 3) how the signal should be used, and 4) in what contexts clicker training should be used. how the signal should be introduced there were two main methods recommended when introducing a new dog to clicker training: 1) conditioning the signal-food pairing, or 2) skipping the pairing and immediately starting to use the signal when reinforcing desired behaviours (figure 3). the first method was the more common of the two methods. sources explained a method called “loading” or “charging” the clicker by which the signal was repeatedly paired with a food reward, essentially building an association between the signal and the subsequent reward. “create a reflexive link between the sound (‘click’) and the primary reinforcer (‘treat’)…the dog learns that click = treat” (book7). during this “loading” process, sources emphasised that the click-treat is not contingent on the dog performing a specific behaviour, but some clarified that “i'm usually looking for the dog not to be doing a behaviour i don't necessarily want” (int9) and only click and treat “as long as your dog isn't doing anything naughty at the moment” (web3). interestingly, there was vast variation in the number of pairing presentations that sources recommended for the dog to develop an association between the signal and reward. many sources did not provide a recommendation, but of those who did, recommendations ranged from “two or three times” (web1) to “one to two hundred reps” (int13). however, int13 clarified that 100-200 repetitions was “a gross overestimation” and that dogs seemed to catch on rather quickly, adding that “3 to 4 can create a classically conditioned response.” in fact, a number of sources followed the second method of introducing a new dog to clicker training. these sources suggested that “loading” the clicker by simply pairing the clicker signal with food is unnecessary. web1 advised to “immediately choose some specific behavior to click” and int3 described success without “loading” the clicker: “i just start clicking and treating. i just teach a very very simple behaviour and they very quickly understand what the clicker means.” this advice suggests that the “loading” procedure is perhaps unnecessary, but rather that dogs may learn the function of the clicker as their experience increases. regardless of the recommended number of repetitions, most sources stated that they had some way of knowing when the dog understood the click-treat association. book5 explained a common test to determine whether the click-treat association was sufficiently “loaded”: “wait for a time during the day when he is not paying attention to you or he is distracted” then click the clicker and “if he looks at you or comes toward you…he is ‘loaded’.” other sources listed behavioural clues such as pairing the click-treat “until the dog begins to look for the treat when he hears the click” (web4) or “until, when you click they look at you… [with] that expectant face” (int6). for sources who chose to introduce the clicker within the context of reinforcing a behaviour, a dog was considered to understand the clicker when “at some point, the learner figures it out and begins offering the behavior ‘on purpose’” (web1). page 9 clicker training: research versus practice creative common license 4.0 – non commercial – share alike – attribution feng et al. figure 3. coded categories for the sub-question: “how should the signal be introduced?” under the main question of “what methods are generally considered ‘best practice’ with relation to clicker training?” https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science somewhat surprisingly, the criteria used in empirical studies for ensuring sufficient click-treat association differ quite considerably from what was mentioned by most of the sources. smith and davis (2008) used a sample of clicker-naïve dogs and reported that “[to] ensure that dogs in the clicker group understood that the click predicted food delivery, they were required to meet the criterion of eating the food from the bowl within 5 s of the click for 10 consecutive trials or to receive a maximum of 20 click-food pairings” (p. 321). this criterion does not exclude the possibility that the dogs merely approached the food once it appeared in the bowl, rather than understanding that the click predicted food delivery. in addition, all dogs, regardless of foodapproaching behaviour, were considered to have been sufficiently conditioned to the click-food pairing after “a maximum of 20 click-food pairings”. in this case, industry recommendations appeared more sophisticated, but perhaps less precise, than methods used in empirical studies. perhaps the precision required by empirical studies has contributed to their failure to demonstrate the efficacy of clicker training, whereas in applied settings individual differences are more easily accommodated. what signals can be used the choice of reward-predicting signal appeared to be a contentious issue. a list of all suggested auditory, tactile, and visual signals identified by the interviewees, books and websites considered in this study is presented in figure 4. some sources firmly believed that the mechanical clicker device was the ideal signal, while others thought that a verbal marker would be best. int9 commented that, “unlike the verbal or other markers, you have to have a clicker with you. it's just a tool that you have to carry around.” sources who supported the clicker listed traits such as: “precise and unambiguous” (book6), “[a] clear, unique and consistent sound” (book5), and “does not convey emotionally loaded approval or disapproval” (web1), while those who supported a verbal marker listed traits such as: “mouldable as far as what you need at a specific time,” “[not] a tool that you have to carry around” (int13), and “easily available” (book1). these differences in desirable characteristics of a signal appear to explain individuals’ signal of choice. for example, some sources heavily emphasised the importance of handlers being “neutral in their training” (int13), thus preferring the clicker over a verbal signal for its “non-emotional” (web5) characteristic. on the surface, this focus on emotional neutrality appears to be in conflict with the fun, partnership-building aspects of clicker training that sources had mentioned as benefits. yet, web1 suggests that for handlers, “the word page 10 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2017 | vol.3 | 1 18 figure 4. coded categories for the sub-question: “what signals can be used?” under the main question of “what methods are generally considered ‘best practice’ with relation to clicker training?” ‘yes’ conveys a sense of social approval…[and] when you can't say ‘yes’ you may feel frustrated and disappointed, and your posture may actually say ‘no!’ the dog feels punished —and immediately the learning slows down or stops.” of course, these claims are based on personal experiences rather than empirical evaluation, but undoubtedly warrant further investigation. one published manuscript and two unpublished master’s theses have compared the efficacy of various signals. chiandetti et al. (2016) compared a clicker+food treatment group to a verbal “bravo”+food treatment group and found no significant differences between the two groups in dogs’ learning or generalizing in a boxopening task. wood (2007) compared a clicker+food treatment group to a verbal “good”+food treatment group. unlike chiandetti et al. (2016), wood reported that the clicker+food treatment group learned more quickly and required fewer reinforcements compared to the verbal “good”+food group. in wood’s nose-target task, the clicker+food group required an average of 36 minutes to complete the task compared to an average of 59 minutes required for the verbal “good”+food group. similarly to chiandetti et al. (2016), blandina (n.d.) found no significant differences between clicker+food and verbal “next”+food groups with regard to criteria level reached in a shaping protocol; however, both of the groups performed worse than the food-only group. in light of these incongruent results, further investigation is required before empirical studies can provide useful recommendations to practitioners on the relative efficacy of different signals. how the signal should be used there were three main categories of advice with regards to how the signal should be used: what to click, how to click, and how to reward after the click (figure 5). in terms of what to click, sources generally agreed that the trainer should pre-define a training criteria (“making sure i have clear criteria” (int3)) and the signal should be used “[the] exact moment the dog does what you want” (web3). there was disagreement, however, as to whether the dog should be required to remain in position after the signal in order to receive the subsequent food reward. “[the] click ends the behaviour” (int8) was a commonly repeated saying. book2 explained this as follows: “we don’t care what the dog is doing when we feed him: only when we click.” in contrast, book5 advised: “if she hears the click and makes a move to come out of her stay, do not give the treat. rather, lure her back into a sit position, and then give the treat.” at present, empirical evidence is unavailable to help determine whether or not choosing to require the dog to maintain the behaviour after the click would impact learning. with regard to how the signal should be delivered, sources emphasised the importance of delivering the page 11 clicker training: research versus practice creative common license 4.0 – non commercial – share alike – attribution feng et al. figure 5. coded categories for the sub-question: “how should the signal be used?” under the main question of “what methods are generally considered ‘best practice’ with relation to clicker training?” https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science signal prior to the primary reinforcement. “click first, then treat” (book5). sources also emphasised “not clicking multiple times per treat” (int3); one correct behaviour, regardless of how good or impressive, should only be followed by one click. most sources also agreed that “you must always reward after a click” (book5). int10 explained that she considers the click “as the promise between me and the dog…if i've clicked and don't give them reinforcement you can really see their disappointment.” in contrast, a few sources argued that a tangible reward was not required after every signal. for example, int2 described the following training situation: “‘click for this, click for this, click for this’ then get the reinforcer at the end.” numerous studies performed with laboratory animals such as rats (e.g. zimmerman 1959; zimmerman and hanford 1966) have demonstrated that bar-pressing behaviours can be successfully trained and maintained when the secondary reinforcement (equivalent to the click) was only followed by primary reinforcement (e.g. food) intermittently or even when secondary reinforcement was not followed by primary reinforcement at all. however, these subjects were conditioned to the predictor signals under food or water deprivation conditions (e.g. holding at 80% freefeeding weight or going 23 hours without water). such deprivation protocols are generally considered unnecessary and unethical for companion animal training (pryor 1999), making direct comparisons difficult. whether or not this or other distinctions between pet dog training conditions and laboratory training conditions affect the function of clicker-type predictor signals is unclear. an unpublished thesis from 2007 began studying the effect of an intermittent ratio of primary reinforcement following a click on learning in dogs. wennmacher (2007) reported “increased noncompliance and other unwanted behaviors”, along with a decreased rate of learning, during a 50% rate of reinforcement (treat after every other click) as compared to a 100% rate of reinforcement (treat after every click). these results support the belief that every click needs to be followed by a reward. however, whether this reward needs to be a food reward is unclear. some sources thought that the reward after the click did not necessarily have to be food and could be praise, petting, or a game of tug instead. web5 advised to “[give]…food one time, then play with a toy, then just rub his ears and praise him.” in fact, in a non-learning context, a recent study using fmri and preference tests suggested that some dogs prefer social praise over food rewards (cook et al. 2016). another study found that dogs preferred petting over praise (feuerbacher and wynne 2015). however, when training a nose-targeting response, praise was found to be a relatively ineffective reinforcer compared to food (feuerbacher and wynne 2012). further research is required to determine whether wennmacher’s findings of decreased rates of learning hold true when the click is occasionally followed by a reward other than food. surprisingly few sources emphasised the importance of a quickly delivered food reward after the signal. web1 suggested “delivering the treat as soon as possible after the click,” however most interviewees noted that the clicker allowed them to be less strict with the timing of their food reward (“the clicker gives me time to fumble and actually get the actual reward” (int6)). a 2015 unpublished dissertation, investigating the effect of delayed positive reinforcement on learning in dogs, reported that an immediate signal followed by a onesecond delayed food reinforcement resulted in lower rates of success than immediate food reinforcement. it was also not significantly different from a one-second delayed signal and food (browne 2015). these results warrant further research, as it is possible that the advice being offered by many practitioners could be inappropriate. in the only peer-reviewed publication on the efficacy of clicker training compared to food alone with dogs, smith and davis (2008) compared a food-only group to a clicker+food group where the food was delivered “approximately 1 s after a click”. they found no differences in rate of learning between dogs who received a click+treat versus treat-only. browne’s finding regarding the impact of a one second delay provide one potential explanation for these null results in smith and davis (2008), who compared a food-only treatment to an immediate signal, 1 second delayed food treatment. overall, the available empirical evidence in dogs appears to support the recommendation that the treat should be delivered as soon as possible after the click, and suggest that page 12 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2017 | vol.3 | 1 18 perhaps the clicker does not in fact “give [the trainer] time to fumble and…get the actual reward” (int6). contexts where clicker training should be used we asked interviewees if there were particular contexts in which clicker training would be expected to be beneficial and collected a wide range of beliefs. some considered clicker training to be appropriate “in all situations” (int11), while others would only use a clicker-type reward predicting signal “in a controlled environment” (int7). others stated that clicker training should be exclusive to professionals: “don’t do it unless you’re a professional” (int1) (figure 6). the majority of sources fell near the middle and found clicker-type training to be beneficial in some, but not all, contexts. in terms of training contexts where clicker training would not be beneficial, multiple interviewees noted that they “would not use it with bat®” (int3) (behaviour page 13 clicker training: research versus practice creative common license 4.0 – non commercial – share alike – attribution feng et al. figure 6. coded categories for the sub-question: “in what contexts should clicker training be used?” under the main question of “what methods are generally considered ‘best practice’ with relation to clicker training?” https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science adjustment training) or for “scent work” (int9). when asked about the contexts in which clicker training was most beneficial, sources listed characteristics such as “activities requiring multiple steps” (book7) (i.e. taught by shaping), “training behaviours at a distance” (int13), and “some sort of physical behaviour” (int1). in addition, across training contexts, most sources emphasised that clicker training was meant to be used when “teaching any new behaviour” (web5) but “once it's a learned behaviour you don't need [the clicker] ” (int9). in particular, multiple sources recommended clicker training when using a method of behaviour generation called “shaping” (book2), where new behaviours are taught by systematically reinforcing behaviour approximations closer and closer to a goal behaviour (yin 2004). while the efficacy of clicker training in a shaping context has not been assessed relative to a food-only control group, three studies (one unpublished thesis and two peer-reviewed articles) have investigated the efficacy of shaping (combined with clicker training) as compared to other methods of behaviour generation. d’onofrio’s (2015) unpublished thesis investigated the efficacy of shaping as compared to simply capturing spontaneous behaviour and found that, for training the tasks of picking up a wallet or medicine bag, shaping was more effective than just waiting for the behaviour to occur. fugazza and miklósi published a pair of manuscripts comparing clicker training (in the context of shaping) to the ‘do as i do’ (topal et al. 2006) training protocol (fugazza and miklósi 2014; 2015). these two studies suggest that, in terms of training time, ‘do as i do’ was superior to shaping/clicker training for more complex and object-related actions; however, these differences were less pronounced for simple behaviours and body movement-related tasks. these findings are interesting and potentially suggest a shift toward training techniques that are more cognitively complex than the trial-and-error learning methods historically associated with clicker training. unfortunately, the extent to which these results can be used to inform judgements regarding the efficacy of the clicker as a reward-predicting signal is limited, as this was not a question directly addressed in the study design. strengths, limitations and suggestions for further research this study used a range of interviews with dog trainers, and also analysed a number of books and websites, in order to describe and evaluate the phenomenon of clicker training. such an approach provided a strong foundational understanding of clicker training; however, there are a few limitations that motivate further investigation. sources were only accessible in english and the majority were from female proponents of clicker training. it is unknown if differences found between sources or source-types are generalisable, and it is possible that not all common opinions have been captured. a larger-scale investigation is required to determine if the results of this study are generalisable to a wider population of dog owners and trainers. the decision to interview self-identified dog trainers rather than only interviewing those with specific certifications or credentials meant that any individual, including those with no formal training, could have been interviewed. unfortunately, there does not exist an international governing body for the certification or regulation of dog trainers that could have provided an unbiased means of restricting participants. the qualitative sampling method used for the interviews obtained a diverse sample of individual beliefs existing in the industry, rather than a sample representative of specific groups in a way that would permit generalisations about industry standard beliefs and practices regarding clicker training (morse 1995). interviewing individuals with varied dog training backgrounds likely contributed to obtaining a broad and varied perspective. the use of google.com database queries allowed the source selection to mimic a likely method by which individuals interested in learning about clicker training might go about locating relevant reading materials. however, this method has a number of potential limitations to its validity and generalisability. first, the database search excludes any sources of advice without an internet presence and could over-represent those sources with better search engine optimisation strategies. secondly, these sources reflect items that the google.com proprietary algorithm deemed most relevant and may not reflect the sources most often page 14 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2017 | vol.3 | 1 18 recommended by industry professionals. finally, this method required the use of keywords specific to clicker training, which resulted in sources that generally recommended its use. as a result, it is unlikely that the content from the sources alone fully represents the beliefs and practices of dog trainers across the industry. specifically, this method of source selection resulted in limited representation of the thoughts and beliefs of dog trainers who choose not to use clicker training. nevertheless, that was not the primary objective of this study, and the relative objectivity of this method of source selection compared to alternative methods outweighed these limitations. evaluation of a wider population of individuals would make it possible to begin determining the relative popularity of certain opinions. in addition, the use of the google.com keyword searches did not permit differentiation between sources directed towards dog owners or trainers. it is reasonable to suspect that sources written for a professional audience would include different materials than those written for first-time dog owners. somewhat fortuitously, however, the results from the search engine query captured both types of sources and, as such, both types of information were represented in the analysis. ultimately, the use of google.com to locate clicker training specific books and websites provided a reasonable starting point for understanding the motivations behind clicker training and common “best practice” recommendations. to compare and contrast the opinions of specific groups or source types, future research requires much larger participant numbers and more directed recruitment of particular sample groups, such as those who are for versus against clicker training, or novice dog owners versus experienced dog trainers. finally, a number of follow-up studies are recommended based on the findings of this preliminary research. it would be beneficial to test the hypothesis that benefits of clicker training are more pronounced (or perhaps only present) in applied contexts when training dogs that have sufficient prior experience with clicker training. empirical assessment of the number of click-treat pairings required for pet dogs to develop the association between the clicker sound and subsequent food is also warranted. furthermore, it would be valuable to know if this click-treat association is required prior to introducing the clicker in a training context. in addition, it would be worthwhile to investigate reported benefits aside from improved rates of task acquisition. for example, the claim that clicker training improves the relationship between the handler and the dog could have important implications by strengthening pet-owner relationships. these and other follow-up studies would begin to elucidate the surprisingly inconclusive results from existing empirical studies, while also informing future methodologies and industry recommendations. 4. conclusion in a recent review, we provided three theoretical accounts of why a device such as a clicker might be expected to enhance learning (feng et al. 2016). it was noted in this review that empirical studies examining the use of such devices in applied settings had so far produced mixed, but largely unfavourable, results. to begin to explain this paradox, the first aim in this study was to describe and evaluate dog training practitioners’ beliefs and perceptions of clicker training relative to empirical studies, so as to better understand the phenomenon as it is practiced in an applied setting. a second aim was to use this information to inform discussion regarding the conflicting evidence on clicker training benefits found both between and within applied and empirical settings. to gather a varied perspective, data were collected from 25 sources across three source types: interviews with dog trainers, popular clicker training books, and websites dedicated to clicker training. these sources were evaluated using a deductive qualitative content analysis process within the framework of three main areas: 1) what clicker training is, 2) why people use clicker training, and 3) ‘best practice’ clicker training recommendations. the results of this study provide a comprehensive description of previously unavailable perspectives in the dog training industry with regards to clicker training. overall, it was evident that many of those practitioners who routinely use clickers or similar devices as training tools, believe strongly that these tools are effective. this may be because clickers are used in practice in different ways than they have been evaluated in available studies. the main findings were page 15 clicker training: research versus practice creative common license 4.0 – non commercial – share alike – attribution feng et al. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science that: the definition of ‘clicker training’ is multidimensional; a wide range of beliefs and perceptions exist in regards to why, how, and in what contexts, dog trainers should incorporate clicker-type training methods into their applied practice; and, there is a marked contrast between sophisticated industry tests and seemingly arbitrary empirical criteria for judging when sufficient predictor signal conditioning has taken place. each of these factors may contribute to differences in clicker training efficacy seen in practice versus empirical assessments. the multi-dimensionality of clicker training suggests that assessing the clicker as a secondary reinforcer, as has been done in scientific literature (as reviewed by williams 1994), may not be reflective of what clicker training entails in an applied setting. likewise, differences in criteria for determining when a dog has had sufficient exposure to the predictor signal for it to function as a learning aid, could have similar implications. these inconsistencies could mean that studies aiming to assess clicker training are not evaluating the phenomenon as used by dog trainers, and as such do not see the same benefits. the varied opinions on how to introduce and use a clicker and the contexts in which they are appropriate may also contribute to differences between practitioner experiences and empirical findings. as such, it is essential that these factors be considered before empirical studies on clicker-type reward predicting signals can confidently be used to inform applied practice. follow-up studies expanding on these initial results are therefore required to further elucidate the mismatch between industry experiences and empirical evidence. 5. acknowledgements this study was carried out with the support of a la trobe university postgraduate research scholarship and a la trobe university full fee research scholarship. 6. references blandina, a.g. n.d. to click or not to click: positive reinforcement methods on the acquisition of behavior. unpublished thesis, university of florida. browne, c.m. 2015. the effects of delayed positive reinforcement on learning in dogs. unpublished thesis, university of waikato. browne, c.m., starkey, n.j., foster, t.m., mcewan, j.s. 2013. delayed reinforcement – does it affect learning? journal of veterinary behavior: clinical applications and research 8: e37-e38, doi: 10.1016/j.jveb.2013.04.039. chiandetti, c., avella, s., fongaro, e., cerri, f. 2016. can clicker training facilitate conditioning in dogs? applied animal behaviour science 184: 109-116, doi: 10.1016/j.applanim.2016.08.006. clayton, a. 2016. opening session. lecture notes from clickerexpo, reno, nevada, january 22, 2016. cook, p.f., prichard, a., spivak, m., berns, g.s. 2016. awake canine fmri predicts dogs’ preference for praise versus food. social cognitive and affective neuroscience, doi: 10.1093/scan/nsw102. d'onofrio, j. 2015. measuring the efficiency of clicker training for service dogs. unpublished thesis, the pennsylvania state university. elo, s., kyngäs, h. 2008. the qualitative content analysis process. journal of advanced nursing 62: 107115, doi: 10.1111/j.1365-2648.2007.04569.x. feng, l.c., howell, t.j., bennett, p.c. 2016. how clicker training works: comparing reinforcing, marking, and bridging hypotheses. applied animal behaviour science 181: 34-40, doi: 10.1016/j.applanim.2016.05.012. feuerbacher, e.n., wynne, c.d. 2015. shut up and pet me! domestic dogs (canis lupus familiaris) prefer petting to vocal praise in concurrent and singlealternative choice procedures. behavioural processes 110: 47-59, doi: 10.1016/j.beproc.2014.08.019. feuerbacher, e.n., wynne, c.d.l. 2012. relative efficacy of human social interaction and food as reinforcers for domestic dogs and hand-reared wolves. journal of the experimental analysis of behavior 98: 105129, doi: 10.1901/jeab.2012.98-105. page 16 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2017 | vol.3 | 1 18 fugazza, c., miklósi, á. 2014. should old dog trainers learn new tricks? the efficiency of the do as i do method and shaping/clicker training method to train dogs. applied animal behaviour science 153: 53-61, doi: 10.1016/j.applanim.2014.01.009. fugazza, c., miklósi, á. 2015. social learning in dog training: the effectiveness of the do as i do method compared to shaping/clicker training. applied animal behaviour science 171: 146-151, doi: 10.1016/j.applanim.2015.08.033. gerritsen, r., haak, r., prins, s. 2013. k9 behavior basics: a manual for proven success in operational service dog training. brush education. gillaspy, j.a., brinegar, j.l., bailey, r.e. 2014. operant psychology makes a splash in marine mammal training (1955–1965). journal of the history of the behavioral sciences 50: 231-248, doi: 10.1002/jhbs.21664. grice, g.r. 1948. the relation of secondary reinforcement to delayed reward in visual discrimination learning. journal of experimental psychology 38: 1-16, doi: 10.1037/h0061016. hilles, s. 2014. to use or not to use? the credibility of wikipedia. public services quarterly 10: 245-251, doi: 10.1080/15228959.2014.931204. langbein, j., siebert, k., nuernberg, g., manteuffel, g. 2007. the impact of acoustical secondary reinforcement during shape discrimination learning of dwarf goats (capra hircus). applied animal behaviour science 103: 3544, doi: 10.1016/j.applanim.2006.04.019. lattal, k.a. 1984. signal functions in delayed reinforcement. journal of the experimental analysis of behavior 42: 239-253, doi: 10.1901/jeab.1984.42-239. lattal, k.a. 2010. delayed reinforcement of operant behavior. journal of the experimental analysis of behavior 93: 129-139, doi: 10.1901/jeab.2010.93-129. mccall, c.a., burgin, s.e. 2002. equine utilization of secondary reinforcement during response extinction and acquisition. applied animal behaviour science 78: 253-262, doi: 10.1016/s0168-1591(02)00109-0. morse, j.m. 1995. the significance of saturation. qualitative health research 5: 147-149, doi: 10.1177/104973239500500201. pryor, k. 1999. don't shoot the dog! the new art of teaching and training. 2 ed. bantam books, new york, new york. skinner, b.f. 1938. the behavior of organisms: an experimental analysis. appleton-century, oxford, england. smith, s.m., davis, e.s. 2008. clicker increases resistance to extinction but does not decrease training time of a simple operant task in domestic dogs (canis familiaris). applied animal behaviour science 110: 318329, doi: 10.1016/j.applanim.2007.04.012. topal, j., byrne, r.w., miklosi, a., csanyi, v. 2006. reproducing human actions and action sequences: "do as i do!" in a dog. animal cognition 9: 355-367, doi: doi 10.1007/s10071-006-0051-6. vegh, e.j., bertilsson, e. 2010. agility right from the start: the ultimate training guide to america's fastestgrowing dog sport. sunshine books, inc. wennmacher, p.l. 2007. effects of click+continuous food vs. click+intermittent food on the maintenance of dog behavior. unpublished thesis, university of north texas. williams, b.a. 1994. conditioned reinforcement: experimental and theoretical issues. the behavior analyst 17: 261-285, doi: williams, j.l., friend, t.h., nevill, c.h., archer, g. 2004. the efficacy of a secondary reinforcer (clicker) during acquisition and extinction of an operant task in horses. applied animal behaviour science 88: 331-341, doi: 10.1016/j.applanim.2004.03.008. wood, l. 2007. clicker bridging stimulus efficacy. unpublished thesis, hunter college, new york, new york. yin, s.a. 2004. how to behave so your dog behaves. tfh publications, c2004. page 17 clicker training: research versus practice creative common license 4.0 – non commercial – share alike – attribution feng et al. https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science zimmerman, d.w. 1959. sustained performance in rats based on secondary reinforcement. journal of comparative & physiological psychology 52: 353-358, doi: 10.1037/h0045807. zimmerman, j., hanford, p.v. 1966. sustaining behavior with conditioned reinforcement as the only responseproduced consequence. psychological reports 19: 391401, doi: 10.2466/pr0.1966.19.2.391. page 18 www pet behaviour science org creative common license 4.0 – non commercial – share alike – attribution 2017 | vol.3 | 1 18 the relevance of affiliative relationships in horses: review and future directions the relevance of affiliative relationships in horses: review and future directions helena costa, sara fragoso, filipa heitor* pet behaviour science | 2019, vol.8, 11 26 doi: 10.21071/pbs.v0i8.11463 helena costa, sara fragoso, filipa heitor centro para o conhecimento animal *avenida dos bombeiros voluntários de algés, 40a, 1495020 algés telephone: +351 919129548 portugal paper review * email: heitor.filipa@gmail.com keywords: affiliative relationships; horses; management; social behaviour; welfare highlights • many studies on horse social behaviour focused on social organization, dominance and aggression, but studies on affiliative relationships are fewer and further research is still needed. • affiliative relationships are a social need and they contribute to the stability of social groups, reproductive success and welfare of horses. • studies on affiliative relationships were conducted on domestic and feral horse populations, on a wide variety of ecological and management conditions, and used different data collection and analysis methods, which make comparisons between studies more difficult. page 11creative commons license 4.0 – non commercial – share alike – attribution 2019 | vol. 8 | 11 26 abstract for many years, studies on horse social behaviour focused mainly on social organization, dominance and aggression. there are comparatively fewer studies on affiliative relationships among horses, despite their impact on the stability of social groups, reproductive success and welfare. we believe that it is important to gain a more complete understanding of this dimension of horse social behaviour and to identify areas of research which need to be addressed in more detail. this review summarizes the existing body of scientific knowledge on affiliative relationships among horses. studies were conducted on a large variety of horse populations and environmental conditions, from feral to domestic horses under different management conditions. moreover, studies conducted to date used different methodologies for data collection and analysis which make meaningful comparisons of their results more difficult. we present their main findings concerning the importance of affiliative relationships for horses and the individual and social factors associated with these relationships. furthermore, we discuss the implications of these findings for management of domestic horses and propose avenues for future studies. we hope this review stimulates further research in this area and may contribute scientific knowledge to improve husbandry practices and horse welfare. http://www.petbehaviourscience.org/ • this review summarizes the main findings concerning the importance of affiliative relationships for horses and the individual and social factors associated with these relationships. • we discuss how the scientific knowledge obtained may contribute to improve husbandry practices and welfare of domestic horses and propose avenues for future research. introduction early studies on horse social behaviour focused mainly on social organization as well as dominance relationships and aggression. there is an extensive body of literature concerning social organization (e.g. klingel 1975; feist and mccullough 1976; berger 1986; feh 1999; linklater et al. 1999), dominance relationships and agonistic behaviours in both feral and domestic horses, as well as their implications for management and housing (e.g. clutton-brock et al. 1976; houpt et al. 1978; houpt and keiper 1982; rutberg and greenberg 1990; van dierendonck et al. 1995; weeks et al. 2000; heitor et al. 2006a). affiliative relationships were not as thoroughly addressed, despite their importance for the cohesion and stability of horse social groups, management and welfare. therefore, we believe that affiliative relationships among horses deserve further study. studies on affiliative relationships conducted to date were performed on different horse breeds and populations, as well as on a large variety of environments and management conditions. these studies used a large variety of data collection and data analysis methods which hinders comparisons between their results and makes it more difficult to extract meaningful information. we believe that comparing the main findings of these research studies would allow us to obtain further information on the adaptive value of affiliative behaviours, the influence of environmental and management conditions on affiliative relationships and the factors that affect their development. the knowledge obtained would be very useful in guiding future research and supporting horse management decisions. the present review intends to summarize and compare the main findings of studies conducted to date on affiliative relationships among feral and domestic horses. we also aim to discuss the implications of these findings for husbandry practices in order to improve the welfare of domestic horses. finally, we identify gaps in knowledge and propose avenues for future research. methods data collection was done in august 2017. we compiled scientific papers by searching for publications on affiliative behaviour in horses using the google scholar search engine with combinations of the following keywords: affiliative, behaviour, conflict, equus caballus, grooming, group, horses, play, relationships and social. papers of empirical or observational research specific to the intended topic and published in peer-reviewed english-language journals were included. we also searched the reference list of papers to identify studies that were missed in the initial search. diversity of subjects and methodology in studies on affiliative behaviour studies on affiliative relationships were conducted on different horse populations (equus ferus caballus) and przewalski horse (equus ferus przewalskii) (kolter and zimmermann 1988; keiper 1988; klimov 1988). the horses lived within a diverse range of environmental and social conditions, from feral populations with minimal human intervention (wells and von goldschmidt-rothschild 1979; kimura 1998; cameron et al. 2009) to managed populations in captivity (kolter and zimmermann 1988; keiper 1988; van dierendonck et al. 1995; weeks et al. 2000). therefore, studies focused on feral and semi-feral horse populations composed of naturally formed groups with occasional introductions and removals of horses, as well as artificially formed groups of domestic horses where group composition was completely determined by humans. some horses had been together for many months or years (van dierendonck 1995; weeks et al. 2000; heitor et al. 2006b; heitor and vicente 2010), while other groups had been formed shortly before the study (araba and crowell-davis 1994; bourjade et al. page 12 www pet behaviour science org creative commons license 4.0 – non commercial – share alike – attribution 2019 | vol.8 | 11 -26 2008). in some studies, stallions were permanent members in breeding groups (kimura 1998; cameron et al. 2009; bouskila et al. 2015), while in others, stallions were present only during the reproductive season (heitor et al. 2006b; heitor and vicente 2010). in some cases, stallions were absent (clutton-brock et al. 1976; van dierendonck et al. 1995; weeks et al. 2000; sigurjónsdóttir et al. 2003; van dierendonck et al. 2004) or existed in very low numbers due to the removal of colts (tyler 1972). sometimes foals were weaned and removed from the group to prevent inbreeding, usually before 1-year old, either both sexes (araba and crowell-davis 1994; weeks et al. 2000; heitor et al. 2006b; heitor and vicente 2010) or only males (kimura 1998). in other cases, foals remained in the group and yearlings as well as two-year-olds were present (wells and von goldschmidt-rothschild 1979; crowell-davis et al. 1986; van dierendonck et al. 1995; sigurjónsdóttir et al. 2003; van dierendonck et al. 2004). most authors evaluated affiliative relationships through a combination of measures of spatial association and affiliative interactions (e.g. van dierendonck et al. 1995; kimura 1998; sigurjónsdóttir et al. 2003; van dierendonck et al. 2004; heitor et al. 2006b; cameron et al. 2009; heitor and vicente 2010). nevertheless, some studies used only spatial association measures (weeks et al. 2000; bouskila et al. 2015). proximity was based on the nearest neighbour (wells and von goldschmidtrothschild 1979; arnold and grassia 1982; crowelldavis et al. 1986; kimura 1998; heitor et al. 2006b; heitor and vicente 2010) or associates, i.e. horses within a given distance of the focal animal (araba and crowell-davis 1994; van dierendonck et al. 1995; weeks et al. 2000; sigurjónsdóttir et al. 2003; heitor et al. 2006b; cameron et al. 2009; heitor and vicente 2010). most commonly, associates were defined as being within two body-lengths of each other, which is purported to correspond to to a horse’s personal space (van dierendonck et al. 1995; sigurjónsdóttir et al. 2003; van dierendonck et al. 2004; cameron et al. 2009; bouskila et al. 2015). some authors recorded only affiliative interactions related to mutual grooming and play (clutton-brock et al. 1976; crowell-davis et al. 1986; araba and crowell-davis 1994; van dierendonck et al. 1995; kimura 1998; sigurjónsdóttir et al. 2003; van dierendonck et al. 2004; rho et al. 2007). other studies also included approaches, follows or friendly contacts (wells and von goldschmidt-rothschild 1979; arnold and grassia 1982; heitor et al. 2006b; cameron et al. 2009; heitor and vicente 2010). when assessing the relationship between affiliative relationships and dominance rank, criteria for assessing dyadic dominance relationships was similar between studies, but the methods for constructing the dominance hierarchy and assigning ranks was not always the same. when studying factors related to affiliative relationships, many authors used simple correlations or two-sample tests (e.g. clutton-brock et al. 1976; crowell-davis et al. 1986; araba and crowell-davis 1994; weeks et al. 2000; rho et al. 2007), while others used also matrix correlation tests (e.g. van dierendonck et al. 1995; sigurjónsdóttir et al. 2003; van dierendonck et al. 2004; heitor et al. 2006b; heitor and vicente 2010), cluster analysis (wells and von goldschmidtrothschild 1979; kimura 1998), principal components analysis (arnold and grassia 1982; van dierendonck et al. 2004), generalized linear models (cameron et al. 2009) or social network analysis (bouskila et al. 2015). given the diversity of environmental and management conditions of horse populations studied to date and the diversity of methods applied, comparisons between studies can only provide suggestive evidence of important variables and associations, which must be later confirmed by experimental studies. bearing this in mind, we present the differences as well as the similarities in the findings of those studies and point out the most relevant patterns that emerged from our analysis. the importance of affiliative relationships horses are social animals. the basic reproductive unit of feral horses is the band, a stable group composed of several mares, their offspring and one or more stallions that defend the mares from other males year round, as is typical of female defense polygyny (salter and hudson 1982; kaseda et al. 1995; linklater et al. 1999; linklater 2000). several hypotheses have been formulated to explain the existence of single-stallion bands and multiple stallion bands. feh (1999) found evidence of cooperation among stallions in bands with page 13 the relevance of affiliative relationships in horses creative commons license 4.0 – non commercial – share alike – attribution costa, fragoso, & heitor https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science multiple stallions. linklater and cameron (2000) rejected cooperative hypotheses and proposed mate parasitism and consort hypotheses as better alternative explanations. young males and females usually leave their natal bands upon reaching sexual maturity (2-3 years old) and form new groups or join existing ones (klingel 1975; salter and hudson 1982; berger 1986; feh 1999) to prevent inbreeding (monard and duncan 1996; monard et al. 1996). females join other bands directly, but males integrate bachelor groups for a few years where they develop social skills which are necessary to maintain a band (hoffmann 1985; mcdonnell and haviland 1995; khalil and kaseda 1998). despite frequent changes in composition within bachelor groups, some long-term associations among males can be developed (salter and hudson 1982). the adaptive value of affiliative relationships horses tend to form stable affiliative relationships in feral and domestic horse groups that include adult mares (e.g. stability over a 3-year period: tyler 1972; van dierendonck et al. 2004; heitor and vicente 2010), but unstable bonds that suffer seasonal changes have also been observed (kimura 1998). stable relationships within bands enhance female reproductive success by reducing stallion harassment and inter-mare aggression associated with band change and male takeovers (berger 1983; kaseda et al. 1995; linklater et al. 1999). long-term bonds between stallions and mares may also improve oestrus detection by stallions (duncan 1980; salter and hudson 1982). in addition, cameron et al. (2009) reported that mares with stable group membership that contributed more for social bonding had higher reproductive success. affiliative relationships among horses are expressed by spending time in proximity and participating in affiliative interactions, such as mutual grooming and social play (tyler 1972; wells and von goldschmidtrothschild 1979; arnold and grassia 1982; kimura 1998; sigurjónsdóttir et al. 2003; van dierendonck and spruijt 2012). horses usually have one or more preferred partners for affiliative relationships (tyler 1972; arnold and grassia 1982; estep et al. 1993; van dierendonck et al. 1995; kimura 1998; sigurjónsdóttir et al. 2003; van dierendonck et al. 2004; bouskila et al. 2015). some horses are more popular than others, as measured by the number of group members that have them as a preferred partner (sigurjónsdóttir et al. 2003). preferred partners for mutual grooming tend to be the same as preferred partners for proximity (cluttonbrock et al. 1976; sigurjónsdóttir et al. 2003; van dierendonck et al. 2004; heitor et al. 2006b), but sometimes these partnerships differ (kimura 1998; gilbert-norton et al. 2004). these findings suggest that, although proximity and mutual grooming are both related to bonds in horses, their functions may be slightly different. maintaining affiliative relationships with particular partners within the group is important for horses, both in bands and bachelor groups, as shown by interventions in social interactions of group members (heitor et al. 2006a; van dierendonck et al. 2009; granquist et al. 2012; schneider and krueger 2012; krueger et al. 2015). interventions in agonistic behaviours seemed to promote group cohesion and to prevent social disruption in a bachelor group of przewalski horses (krueger et al. 2015). interventions in affiliative interactions seemed to function as a means to prevent a potential weakening of a horse’s own bonds with preferred partners (van dierendonck et al. 2009, schneider and krueger 2012). in addition, there is some preliminary evidence that affiliative interactions exchanged between horses after a conflict could function as reconciliation behaviours, thereby preventing further aggression and maintain the affiliative relationship between former opponents (cozzi et al. 2010). it has been suggested that affiliative relationships may provide other indirect benefits to mares by reducing aggression received (cameron et al. 2009). aggression increases stress levels and decreases body condition, thereby reducing reproductive success (linklater et al. 1999). nevertheless, frequency of agonistic interactions is not always lower among preferred partners, both among adults (clutton-brock et al. 1976; weeks et al. 2000; heitor et al. 2006b) and among foals (araba and crowell-davis 1994). page 14 www pet behaviour science org creative commons license 4.0 – non commercial – share alike – attribution 2019 | vol.8 | 11 -26 benefits associated with affiliative interactions there are benefits associated with performing affiliative interactions, such as mutual grooming and play. mutual grooming enables horses to obtain care of the coat in areas of the body difficult for them to reach (tyler 1972), promotes appeasement (feist and mccullough 1976) and reduces social tension (feh and de mazières 1993). this behaviour is performed by horses of all sex-age classes but its frequency varies greatly (clutton-brock et al. 1976; keiper 1988; heitor et al. 2006b). in some groups mutual grooming was relatively frequent (sigurjónsdóttir et al. 2003; van dierendonck et al. 2004), but in other groups the frequency was low and some horses were never observed participating in it (wells and von goldschmidt-rothschild 1979; arnold and grassia 1982; crowell-davis et al. 1986; heitor et al. 2006b; heitor and vicente 2010). mutual grooming frequencies vary seasonally, with higher frequencies during spring and summer due to coat shedding and higher density of tabanid flies (tyler 1972; wells and von goldschmidt-rothschild 1979), which could partly account for the variation in study results. play behaviour in horses was described by mcdonnell and poulin (2002) and it is generally believed to improve motor, cognitive and social skills (bekoff and allen 1998). social play is mainly performed by younger horses and males but it is rarely seen in adult mares (tyler 1972; wells and von goldschmidt-rothschild 1979; sigurjónsdóttir et al. 2003; zharkikh and andersen 2009). van dierendonck and spruijt (2012) argued that mutual grooming and play could be considered “ethological needs” because these behaviours are performed by all individuals, self-rewarding, have a rebound effect and chronic stress is induced in absence of a social partner. in addition, van dierendonck and spruijt (2012) reviewed evidence that horses that are deprived from social contact with other horses can suffer chronic stress and engage in abnormal behaviours such as stereotypies. for example, van dierendonck (2006) observed that horses housed in a social-contact-at-adistance system conspicuously anticipated opportunities for direct physical contact with other horses and showed stress reactions when social contact was no longer allowed. consistent with these findings, visser et al. (2008) observed that stress-related behaviours and stereotypies were displayed more frequently in individually housed horses than in horses kept in pairs. in addition, hartmann et al. (2012) reviewed evidence that the opportunity for social contact is related to improved development of social skills, decreased reactivity and decreased aggression in domestic horses. housing young horses singly does not give them the opportunity to practice their social skills, so they are more prone to injuries when interacting with other horses as adults (søndergaard and christensen 2007). in sum, affiliative relationships promote group stability, which is related to increased reproductive success in feral horses. moreover, the opportunity to engage in affiliative interactions is an ethological need for horses and it is necessary for normal development of their social skills. factors associated with affiliative relationships although the importance of affiliative relationships for horses is undeniable, factors that determine the strength of affiliative relationships and choice of preferred partners are less clear. we found contradicting results between studies, which could be due to the differences in horse breeds, management conditions, data collection and data analysis methods of studies conducted to date. we present the main findings of studies that assessed the association between affiliative relationships and the factors age, gender, dominance, kinship and familiarity, reproductive state and social environment. age many authors observed that horses formed stronger bonds with other horses of similar age, especially within their age class (e.g. foals, subadults, adults). this could be due to their similar environmental and social needs. when integrating new bands after natal dispersal, young females usually had one particular subadult female as preferred partner for proximity and affiliative interactions, which was close in age (monard and duncan 1996). in groups without mature stallions, horses more often performed affiliative interactions page 15 the relevance of affiliative relationships in horses creative commons license 4.0 – non commercial – share alike – attribution costa, fragoso, & heitor https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science (mutual grooming and play) and maintained proximity with horses of similar age (clutton-brock et al. 1976; sigurjónsdóttir et al. 2003). in a camargue horse herd with a mature stallion, wells and von goldschmidtrothschild (1979) also observed that, excluding close kin relationships via the mother, horses spent more time in proximity to other horses of similar age. in foals, mutual grooming was most common with other foals (crowell-davis et al. 1986; rho et al. 2007). in addition, when mothers and close relatives were excluded, foals were more often nearest neighbours with other foals (crowell-davis et al. 1986) or partners for affiliative interactions (wells and von goldschmidtrothschild 1979). nevertheless, some exceptions were found: age similarity was not related to affiliative relationships among adult sorraia mares (heitor et al. 2006b) and konik horses (bouskila et al. 2015). studies show that foals and subadults take part in affiliative interactions more frequently than adults, particularly social play (tyler 1972; feist and mccullough 1976; wells and von goldschmidtrothschild 1979; sigurjónsdóttir et al. 2003; zharkikh and andersen 2009) and mutual grooming (keiper 1988; granquist et al. 2012). among adult horses, age seems less important, as it was not associated with frequency of mutual grooming in highland pony mares (clutton-brock et al. 1976) or frequency of affiliative interactions in bachelor przewalski males (zharkikh and andersen 2009). subadults had more preferred partners for mutual grooming than adult mares in harems (granquist et al. 2012). however, the strength of affiliative relationships and diversity of partners was not related to age in adult sorraia mares (heitor and vicente 2010). consistent with these findings, the number of preferred partners for affiliative interactions (mutual grooming and play) and popularity as a partner were not related to age in icelandic horses (sigurjónsdóttir et al. 2003). therefore, studies in horses have not shown a decline in sociability with aging such as referred in humans and nonhuman primates (pavelka 1991; veenema et al. 1997). nevertheless, the effects of aging on affiliative relationships would be best examined by longitudinal studies to allow the control of confounding variables. we do not know of any longitudinal study conducted to date that has investigated the changes in affiliative relationships of horses as they age. gender icelandic horses spent more time in proximity and groomed more often with horses of the same sex (sigurjónsdóttir et al. 2003). araba and crowell-davis (1994) observed that foals associated more often with foals of the same sex than with foals of the opposite sex, both before and after weaning. nevertheless, weeks et al. (2000) reported that the most common spatial associate of foals was not related to gender either before or after weaning. crowell-davis et al. (1986) found that male foals groomed almost exclusively with female foals, fillies groomed other foals irrespective of gender but they were more likely to have other fillies as nearest neighbours. foals were as likely to mutual groom with a foal of the opposite sex as with a foal of the same sex in jeju ponies (rho et al. 2007). young males preferred to play within their own sex-age class, while the subadult females played with both sexes (sigurjónsdóttir et al. 20003). monard et al. (1996) reported that before natal dispersal, young camargue mares played more often with other immatures of both sexes. therefore, it seems that young males tend to mutual groom more often with opposite sex partners and more often play with same-sex partners, while females do not display gender-related differences. female foals took part in mutual grooming more often than male foals in welsh ponies (crowelldavis et al. 1986). young males played more often than females (wells and von goldschmidt-rothschild 1979; sigurjónsdóttir et al. 2003), and they also had more playing partners and were more popular as play partners than females (sigurjónsdóttir et al. 2003). these differences between genders may reflect different functional benefits of affiliative relationships for stallions and mares. for mares, affiliative relationships established with both the band stallion and other mares contribute to stable group membership which increases reproductive success (linklater et al. 1999). for males, bonding with other stallions may be less important because bachelor bands are unstable and the greatest reproductive success is achieved through relationships established with mares in a band (linklater et al. 1999). nevertheless, social play may be more frequent in males than females because a male’s play-fighting experiences at an early age could help develop skills which will be important for stallions in acquiring, page 16 www pet behaviour science org creative commons license 4.0 – non commercial – share alike – attribution 2019 | vol.8 | 11 -26 maintaining and defending a harem from other males (rho et al. 2007). further study is necessary to assess whether mutual grooming and social play at an early age affect the strength of affiliative relationships and reproductive success later in life. dominance relatively stable and linear dominance hierarchies based mainly on age, are usually developed within horse social groups (e.g. clutton-brock et al. 1976; wells and von goldschmidt-rothschild 1979; van dierendonck et al. 1995; sigurjónsdóttir et al. 2003; gilbert-norton et al. 2004; heitor et al. 2006a), although feist and mccullough (1976) found no consistent dominance hierarchy among mares in feral harem groups. contribution to affiliative relationships may be related to dominance relationships within each pair of horses. the dominant individual within each pair of horses has been reported to initiate affiliative interactions more frequently than the subordinate (clutton-brock et al. 1976; wells and von goldschmidt-rothschild 1979; heitor et al. 2010). however, tyler (1972) noted that mutual grooming bouts were most often initiated by subordinates (tyler 1972). subordinates may be inhibited from initiating affiliative interactions with dominants (wells and von goldschmidt-rothschild 1979) due to the increased probability of receiving agonistic interactions from them (heitor et al. 2006a). moreover, subordinates may leave dominants due to agonistic interactions received from them, thereby contributing less to proximity (heitor and vicente 2010). some studies found that horses with similar dominance rank developed stronger affiliative relationships (clutton-brock et al. 1976; wells and von goldschmidtrothschild 1979; kimura 1998; sigurjónsdóttir et al. 2003), spent more time in proximity (van dierendonck et al. 1995; kimura 1998; heitor et al. 2006b) or groomed more often (clutton-brock et al. 1976; sigurjónsdóttir et al. 2003). however, other studies did not find relationships between mutual grooming and rank distance (van dierendonck et al. 1995; van dierendonck et al. 2004; heitor et al. 2006b) or between proximity and rank distance (van dierendonck et al. 2004; bouskila et al. 2015). mutual grooming frequency was not related to rank in free-ranging highland ponies (clutton-brock et al. 1976). nevertheless, lower-ranking przewalski horses in captivity were involved in mutual grooming significantly more often (keiper 1988). popularity as a partner for mutual grooming or play was not related to rank in icelandic horses (sigurjónsdóttir et al. 2003). in addition, rank was not related to sociability (based on the number of spatial associates) in mares and foals (weeks et al. 2000) or to the number of play bouts initiated or terminated by foals (araba and crowelldavis 1994). in sum, affiliative relationships may be influenced by dyadic dominance relationships and rank similarity, but dominance rank seems less important. kinship and familiarity in feral horses’ natal bands, females bond more strongly with their mother and siblings (tyler 1972; wells and von goldschmidt-rothschild 1979; monard et al. 1996). these bonds are usually broken at the time of natal dispersal (klingel 1975; salter and hudson 1982; waring 1983; berger 1986; feh 1999) but longterm bonds may develop among adult matrilineal relatives if they ever meet again (tyler 1972; monard and duncan 1996). some authors found that mares who were close relatives via the mother spent more time in proximity (gilbert-norton et al. 2004) or participated in mutual grooming more often (keiper 1988; sigurjónsdóttir et al. 2003; van dierendonck et al. 2004; heitor et al. 2006b). by contrast, other studies found no relationship between mutual grooming and kinship via the mother (clutton-brock et al. 1976; van dierendonck et al. 1995). moreover, affiliative relationships were not stronger between close relatives in sorraia mares (heitor et al. 2006b; heitor and vicente 2010) or icelandic horses of all age-sex classes without intact stallions (van dierendonck et al. 1995). in herds where all horses were familiar to each other, affiliative relationships were sometimes related to kinship beyond close relatives via the mother, as measured by the degree of genetic relatedness (sigurjónsdóttir et al. 2003; van dierendonck et al. 2004; page 17 the relevance of affiliative relationships in horses creative commons license 4.0 – non commercial – share alike – attribution costa, fragoso, & heitor https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science heitor et al. 2006b). nevertheless, the relationship between kinship and affiliative relationships among mares reported by heitor et al. (2006b) was no longer significant in later years (heitor and vicente 2010). moreover, bouskila et al. (2015) found no significant relationship between genetic relatedness and strength of spatial associations in semi-feral konik horses. monard and duncan (1996) reported that young dispersing females more often joined bands with familiar subadult females that had previously belonged to their maternal groups and they formed close affiliative relationships with them, at least initially. however, in their study, familiar females were often close relatives, so it was not possible to separate the familiarity and kinship effects. arnold and grassia (1982) observed that mares that had been in the same group before the study spent more time close to each other when resting. in icelandic horses, the relationship between frequency of mutual grooming and kinship was not significant when familiarity was controlled for (van dierendonck et al. 2004). van dierendonck et al. (2004) stated that, when unfamiliar animals were introduced into the group, familiarity was a more powerful predictor of the frequency of mutual grooming than kinship. these findings suggest that horses may be able to recognize close kin via the mother and may also be able to discriminate genetic relatedness to a certain extent, although these abilities need to be tested experimentally. affiliative relationships with kin seem more important in natal bands between motheroffspring and siblings than among adults. familiarity with another horse seems to have greater influence on affiliative relationships when animals move to new groups where most other horses are unfamiliar to them. we suggest that selective pressure for developing affiliative relationships based on kin and familiarity may be low in horses because they are unlikely to find close relatives and familiar individuals in their new groups after natal dispersal. reproductive state foaling leads to changes in affiliative relationships among mares (estep et al. 1993; van dierendonck et al. 2004; heitor and vicente 2010). the distance of new mothers to other group members increased after foaling (tyler 1972; klimov 1988; estep et al. 1993; van dierendonck et al. 2004; heitor and vicente 2010). mothers seemed to be mainly responsible for this because they did not receive lower frequency of affiliative interactions (heitor and vicente 2010) but they initiated affiliative interactions less often and contributed less to proximity with group members after foaling (estep et al. 1993; heitor and vicente 2010). estep et al. (1993) also observed that preferred partners for proximity changed after foal birth. mares spent more time in proximity to others in the same reproductive state, so barren mares and mares with foals were spatially separated into sub-groups (arnold and grassia 1982; van dierendonck et al. 2004; heitor and vicente 2010; bouskila et al. 2015). despite changes in spatial proximity, patterns of affiliative interactions suggest that affiliative relationships among mares before foaling are not broken and replaced by new relationships with mares in the same reproductive state. in icelandic horses, mares groomed more often with others in the same reproductive state but they maintained their preferred mutual grooming partners (van dierendonck et al. 2004). in sorraia horses, mares did not engage in affiliative interactions more frequently with others in the same reproductive state (heitor and vicente 2010). social isolation after parturition is important for imprinting and individual recognition between dam and foal, preventing foals from bonding with other horses in the first few days after birth (estep et al. 1993; van dierendonck et al. 2004). the sub-grouping of dams and foals may facilitate protection of those foals from interactions with other group members (van dierendonck et al. 2004). in addition, mutual attraction between mares with foals may be a by-product of mutual attraction between foals (van dierendonck et al. 2004). as foals begin to interact with each other and their dams maintain proximity to them, mares in the same reproductive state may end up spending more time in proximity. social environment sigurjónsdóttir et al. (2003) noticed that mutual grooming seemed to be more common in groups page 18 www pet behaviour science org creative commons license 4.0 – non commercial – share alike – attribution 2019 | vol.8 | 11 -26 without stallions. in addition, in these groups mares more often groomed with other mares of similar age (tyler 1972; clutton-brock et al. 1976; sigurjónsdóttir et al. 2003) while mares in harems engaged in affiliative interactions more frequently with their 0-3 year old offspring but rarely with other mares (wells and von goldschmidt-rothschild 1979). granquist et al. (2012) observed that in stable groups with a stallion, mares showed less developed dominance hierarchies, lower frequencies of aggression, fewer preferred mutual grooming partners and made fewer interventions in affiliative interactions of group members than in groups without stallions. the overall mutual grooming frequency was similar in harems and non-stallion groups. granquist et al. (2012) suggested that the presence of stallions and their herding movements to maintain cohesiveness of the group may reduce the need or opportunity for interactions among mares (granquist et al. 2012). moreover, sigurjónsdóttir et al. (2003) suggested that affiliative relationships could be dependent on the dominance status of the stallion and that the social structure of groups without a stallion could be similar to that of harems with a low-ranking stallion. affiliative relationships among mares were also deeply affected by stallion behaviour in a group of przewalski horses in captivity: the stallion directed intense aggression towards some of the mares and herded other mares away, causing splitting of the group (kolter and zimmermann 1988). mares affect the choice of preferred partners of their offspring before weaning and also to some extent after weaning, although less significantly so. foals and yearlings associate more with the offspring of their dam’s preferred spatial associate (wells and von goldschmidt-rothschild 1979; araba and crowelldavis 1994; weeks et al. 2000). nevertheless, the stability of this influence in the long term was not yet addressed. heitor and vicente (2008) reported that rank and maternal experience of mares had little influence on the development of affiliative relationships of their foals with other horses. sociability rates based on the number of spatial associates were significantly correlated in dams and their foals prior to weaning, but not after weaning (weeks et al. 2000). the presence of adults beyond the mother and the adult-young ratio are important for social development of young horses. bourjade et al. (2008) found that when adults were present in same-sex groups, young horses between 1-2 years old had clear preferred partners, displayed new behaviour patterns and showed decreased aggression, compared with same-sex groups without adults. in groups with lower adult-young ratio, young przewalski horses between 1-2 years old spent more time in association with other young horses, segregated more from adults and were more aggressive than in higher adult-young ratio groups (bourjade et al. 2009). nevertheless, the frequency of affiliative interactions and number of preferred spatial partners was not related to the adult-young ratio (bourjade et al. 2009). because dams and other adults can be important learning models for young horses (bourjade et al. 2009), more studies are needed to address the consequences of management procedures such as weaning and keeping weaned young horses in same-age groups on the development of social skills and affiliative relationships in horses. implications for husbandry practices and welfare broom (1986) defined welfare as the state of an animal with regards to its attempts to cope with its environment. domestic horses do not have control over some features of their environment (e.g. home range, group members and mating partners). therefore, understanding the ethological needs of horses regarding affiliative behaviour and relationships may provide valuable information to improve husbandry practices and horse welfare. domestication caused changes in horse behaviour, especially decreased fear and reactivity, but did not significantly affect social behaviour (van dierendonck and spruijt 2012). as stated by these authors, engaging in affiliative interactions such as mutual grooming and social play is an ethological need. although husbandry conditions have improved over the last decades, many domestic horses are still housed individually in enclosed stables with limited space available, most of them in boxes (hartmann et al. 2012). physical contact with other horses is typically limited, especially among mature stallions (hartmann et al. 2012). because they are deprived of social contact, most affiliative behaviours cannot be performed. in addition, page 19 the relevance of affiliative relationships in horses creative commons license 4.0 – non commercial – share alike – attribution costa, fragoso, & heitor https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science whereas horses kept at pasture tend to associate with preferred group members and distance themselves from others, social relationships with a stabled horse’s closest neighbours are not often taken into account when deciding on housing arrangements (redgate and davidson 2007). in other cases, horses are (semi-)permanently housed in large groups. confined spaces where animals cannot avoid dominant individuals and aggressive behaviours may result in more defined dominance hierarchies, increased competition for resources or higher aggression rates (houpt and keiper 1982; keiper 1986; mills and nankervis 1999; price 1999; andersen et al. 2006). in addition, in managed horse herds where group composition is frequently determined or changed by man, bonds may not develop or be disrupted (tyler 1972) and relationships need to be readjusted periodically, which may cause increased aggression (waring 1983). based on the main findings of this review, we propose some recommendations regarding management and husbandry of domestic horses in order to improve their welfare. individual versus group housing the findings of this review concerning the importance of affiliative relationships provide a strong basis for recommending the use of housing conditions where horses are allowed permanent social contact and development of affiliative relationships. horses should be kept as a group at pasture or with access to an outdoor paddock. the enclosure area will depend on group size, but it should provide them ample space to move away from other horses if needed. this is important, for example, for new mothers to create some distance between their newborn foal and other group members. group composition horses tend to form stronger bonds with others of similar age, especially among young horses. moreover, young horses get frequently involved in affiliative interactions with partners of both sexes. the presence of adults beyond the mother is important for the development of social skills, especially for young horses, because adults may serve as role models. therefore, it seems that young horses would benefit most from being kept in groups with different age-sex classes for as long as possible, instead of the common practice of weaning foals before 1-year old and keeping them in same-age groups. however, our findings show that it is generally difficult to predict whether horses will form strong affiliative relationships based on individual factors such as those assessed in this review (e.g. age, gender, dominance, kinship) because these factors were not consistently related to affiliative relationships across different horse populations. when decisions have to be made on which horses will be grouped together, it may then be more useful for horse keepers to monitor affiliative relationships between horses than trying to predict these relationships on the basis of individual factors. observing social behaviour will allow them to make the necessary adjustments in group composition, housing conditions and management procedures, so that welfare is improved. this monitoring is especially important after introduction or removal of horses and after the birth of foals to assess the effects on affiliative relationships and prevent possible cases of increased aggression. this review presents some behavioural measures that may be used by horse keepers for this purpose. introduction and removal of horses from groups because horses develop strong and stable affiliative relationships which affect their welfare, changes in group composition due to introduction and removal of horses should be avoided as much as possible. when animals moved to new groups where most other horses were unfamiliar to them, familiarity was an important factor related to the development of affiliative relationships. therefore, when a horse is to be transferred to a new group, choosing a group with at least one familiar horse should be preferred, if one is available. gaps in knowledge and avenues for future research considering the findings of previous studies, we identified areas of research which could be addressed page 20 www pet behaviour science org creative commons license 4.0 – non commercial – share alike – attribution 2019 | vol.8 | 11 -26 in more detail in order to gain a more complete understanding of affiliative relationships among horses. here we present themes which deserve further investigation and suggest improvements considering methodological issues. differences in personality, life history, social competence and social learning may cause interindividual variation in the way each horse expresses an affiliative relationship. for example, as mentioned before, some horses were never observed performing mutual grooming but expressed other types of affiliative behaviour. therefore, we believe it is important to apply statistical tests that take individual variation into account and to use multiple behavioural measures to assess affiliative relationships. we recommend the use of several types of affiliative interactions (e.g. approach, follow, friendly contacts, mutual grooming) and proximity measures (e.g. associates, nearest neighbours) and the assessment of correlations between these measures. for the study of proximity relationships, biologically meaningful information can best be obtained by evaluating which individuals are within the personal space (van dierendonck et al. 2004) because this is the area immediately around the horse in which only close companions are tolerated (for review see mills and nankervis 1999). proximity measures could be more relevant if they take into account a horse’s personal space as well as the enclosure or pasture area weighted by the number of horses. the terminology for describing affiliative relationships could be made more objective by using descriptive terms. for example, some authors used the term “preferred partners”, which suggests that affiliative relationships were based on a cognitive ability to make a choice based on preference. this cognitive feature was not tested in observational studies. in addition, there may be social constraints that prevent horses from associating with certain group members thereby imposing limits on their choices. therefore, “most common partners” or other descriptive terms would be more adequate. although affiliative relationships are commonly assessed through a variety of behavioural measures, social skills have not been objectively defined and measured in horses. more study is also needed to understand how the development of these skills is affected by maternal investment, age at weaning, group members (e.g. peer number, age and gender) and social play. future studies could also address the impact of social skills on reproductive success later in life. the underlying motivations and functions of interference and reconciliation behaviours among horses and their role in affiliative relationships could be studied in greater detail. regarding interference behaviours, it is important to analyze the type of behaviour that was used to interfere (affiliative or agonistic), the identity of the target horse and the immediate effect of interference (e.g. separation, replacement). regarding reconciliation, we believe that the type and intensity of agonistic interactions and the kind of affiliative interactions exchanged between the horses after the agonistic interaction should be taken into account. moreover, when studying interference and reconciliation, social interactions may be directed to group members that are interacting with one another merely by chance. for example, horses that intend to interfere in interactions of group members may be expected to pay attention to those social interactions prior to intervening on them. therefore, attention could also be measured, through gazing, ear turning, head and neck lifting and orientation of the body towards the target. cooperative behaviours and their association with affiliative relationships have not been investigated in horses. cooperation in horses has been given little attention, except among males in bands with multiple stallions, as referred earlier. heitor et al. (2011) reported that sexual interference behaviours in sorraia horses seemed to be related to mare protection, but more studies are needed to understand whether these behaviours could be explained by cooperation. it could also be tested whether interference in agonistic interactions of other mares or their foals can be explained through cooperative hypotheses. vigilance behaviours and tolerance at feeding sites could also be investigated as subtle forms of cooperation. conclusions horses develop strong and stable affiliative page 21 the relevance of affiliative relationships in horses creative commons license 4.0 – non commercial – share alike – attribution costa, fragoso, & heitor https://es.pinterest.com/petbehavioursci https://twitter.com/petbehav_sci https://www.facebook.com/petbehaviourscience https://www.linkedin.com/company/pet-behaviour-science relationships that have been favoured by natural selection through increased survival and reproductive success. affiliative relationships are associated with a variety of individual and social factors, but the relative importance of these factors shows large variation between horse populations. age similarity, mare reproductive state and social environment were some of the most relevant factors associated with affiliative relationships. we argue that horse husbandry practices should be supported by the body of scientific knowledge that has been gathered to date on horses’ social needs and affiliative behaviour. domestic horses should be provided with conditions to express their natural affiliative behaviours and develop social skills. regarding fundamental research, we suggest that promising areas for future study include social skills, interference, reconciliation and cooperative behaviours. ethical statement the work described in this article did not require approval by eu directive 2010/63/eu for animal experiments. conflict of interest statement we declare to have no conflict of interest. references 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