Highlights in BioScience ISSN:2682-4043 DOI:10.36462/H.BioSci.202109 Research Article Open Access 1 Department of Microbiology, Faculty of Vet- erinary Medicine, Adnan Menderes University, 09016, Isikli, Aydin, Turkey. 2 Department of Microbiology, Institute of Health Sciences, Adnan Menderes University, 09016, Efeler, Aydin, Turkey. Contacts of authors * To whom correspondence should be addressed: uparin@adu.edu.tr Received: March 11, 2021 Accepted: June 15, 2021 Published: October 10, 2021 Citation: Kirkan Ş, Parin U, Çapakçioglu H . Iden- tification of pathogen bacteria from camel (Camelus dromedarius) mastitis and investigation of antibi- otic susceptibility. 2021 Oct 10;4:bs202109 Copyright: © 2021 Kirkan et al.. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Data Availability Statement: All relevant data are within the paper and supplementary materials. Funding: This research was funded by Aydın Adnan Menderes University Scientific Committee by grant number VTF-18017. Competing interests: The authors declare that they have no competing interests. Identification of pathogen bacteria from camel (Camelus dromedarius) mastitis and investigation of antibiotic susceptibility Şükrü Kirkan1 ><, Uğur Parin*1 ><, Halil Çapakçioglu2 >< Abstract The scope of this study was to investigate the presence of pathogenic bacteria in milk from female camels with mastitis and to select antibiotics for treatment with antibiotic susceptibility testing. A total of 40 milk samples taken from 20 dromedarian females, after application of CMT test and determination of SCC values, the camels were diag- nosed with subclinical mastitis. Milk samples were inoculated into blood agar for iden- tification of bacterial agents leading to mastitis. A total of 4 (12.5%) Staphylococcus aureus, 4 (12.5%) S. auricularis, 2 (6.25%) S. pettenkoperi, 2 (6.25%) S. cohnii spp. cohnii, 2 (6.25%) S. equorum, 2 (6.25%) S. capitis, 2 (6.25%) Streptococcus agalac- tiae, 2 (6.25%) S. dysgalactiae, 4 (12.5%) Escherichia coli, 2 (%) 6.25) Pseudomonas pseudalcaligenes, 2 (6.25%) Corynebacterium pseudotuberculosis, 2 (6.25%) Aerococ- cus viridans and 2 (6.25%) Gemella morbillorum were identified. Gram-positive bacteria were sensitive to Levofloxacin, Linezolid and Tetracycline and Daptomycin, resistant to Beta lactam-group antibiotics and macrolides. Vancomycin resistance was determined in S. aureus and S. cohnii spp. cohnii strains. Gram-negative strains are found generally susceptible to Cefepime and Pipersilin; resistant to Trimethoprim-sulfomethoxazole and Amoxicillin-Clavulanic acid. As a result, it is recommended to use antibiotic use to pre- vent the development of antimicrobial resistance as well as mastitis control methods such as the prevention of infection and monitoring the health status of the mammary of camels. Keywords: Camel, Subclinical mastitis, Identification, Antimicrobial susceptibility. Introduction Camel milk, meat and products have appeared in many cuisines around the world for centuries. Camel milk has beneficial effects on many biological processes such as digestion, absorption, growth and immunity [1]. Additionally, camel milk can be stored at room temperature longer than milk from other animals [2]. Camel whey protein contains a heterogeneous group of proteins, including serum albumin, α-lactoalbumin, immunoglobulin, lactoferrin, and peptidoglycan recognition protein [3]. Compared to milk, camel milk contains more antibacterial substances and has a high concentration of vitamin C, so it has valuable nutritional properties. Milk is characterized by a high percentage of lactoferrin, which can be considered a good source of minerals and vitamins. Also, because camel milk has the most important nutrients, camel milk can meet most of human’s daily needs in these nutrients. Camel milk has a similar composition to human milk due to its protective proteins such as low cholesterol, low sugar, high mineral content (sodium, potassium, iron, copper, zinc and mag- nesium), vitamin C, lactoferrin, lactoperoxidase, immunoglobulins and lysozyme [4]. The present studies confirmed that camel milk is unique in terms of antioxidant, antibacterial, antiviral, anti- fungal, anti-hepatitis, tuberculosis, hypoglycemic, anti-cancer, anti-tumor, anti-aging, auto-immune disease, cosmetic and detergents [5-6]. Mastitis is a major global complex disease among dairy live- stock with high economic losses. Camel mastitis is estimated to affect more than 25% of lactating camels. It is also known to cause about 70% loss from milk production. Mastitis has both extreme animal infections and economic significance. It is responsible for some harmful effects for human health and animal production. There is little published information about the pathogens associated with camel mastitis compared to bovine mastitis. Highlights in BioScience Page 1 of 7 October 2021|Volume 4 https://doi.org/10.36462/H.BioSci.202109 https://creativecommons.org/licenses/by/4.0/ skirkan@adu.edu.tr https://orcid.org/0000-0001-5111-8656 uparin@adu.edu.tr https://orcid.org/0000-0002-0788-5708 halilcapakcioglu@hotmail.com http://bioscience.highlightsin.org/ Kirkan et al., 2021 Identification of pathogen bacteria from camel (Camelus dromedarius) mastitis However, much detailed research still remains to be accom- plished, especially in the mastery of breeding techniques, since these species is confronted with particular health issues, includ- ing mastitis, which can be significant. There has long been a lack of concern in camel mastitis on the grounds that clinical mastitis in this species is infrequent. Bacterial infection can be a major cause of mastitis in livestock. Several bacterial agents are associated with mastitis cases and their presence in milk can have a negative impact on consumer health [8-9]. Some research shows the genus Staphylococcus sp., Streptococcus sp., [10,11], Micrococcus sp., Streptococcus agalactiae, Coagulase Negative Staphylococci [12], Staphylococcus epidermidis, Mannheimia haemolytica, Escherichia coli and Corynebacterium sp. were isolated from camel mastitis cases. Camel has been linked to the cause of mastitis. Mammary infections (mastitis) are the leading risks of herd management in camel breeding as well as in cattle breeding. The scope of this study was to investigate the presence of pathogenic bacteria in milk from female camels with mastitis and to select antibiotics for treatment with antibiotic susceptibil- ity testing. Materials and Methods Sample collection Milk samples were taken from 20 female one-humped camels suspected of having mastitis during lactation. The study was car- ried out from August 2018 to April 2019 at private camel farms in Aydin province, Turkey. In this study, 40 camel milk sam- ples were obtained from the right and left lobes of 20 dromedary camels at farms with a capacity of 5 to 20 camels. Animal material consists of female camels aged 4 to 10 years who are lactating without antibiotic treatment. Mastitis was diagnosed by applying CMT (California Mastitis Test DeLaval®, Sweden) and determining somatic cell counts. NucleoCounter™ SCC- 100 (Chemometec®, Denmark) was used to determine the SCC value. The identification of bacterial isolates was determined by an automatic identification system Phoenix™ M50 (Becton, Dickinson U.K. Limited). Ethics of animal use The authors of this research hereby declare that collection of specimens was carried out in accordance with the guidelines laid down by the US National Guide for the Care and Use of Labora- tory Animals, National Academy Press, Washington, D.C. 2011 and with the European Communities Council Directive of 24 November 1986 (86/609/EEC). California mastitis test Somatic cell number score in milk samples was determined qualitatively. Detergent containing bromocresol purple was used as the reagent to release the nucleic acid and rupture the somatic cell membrane, and it was observed whether a gel-like matrix was formed whose viscosity was proportional to the leukocyte number. Plastic shovels with four glasses were used. Equal amounts (3 ml) of milk and reagent were put into each glass of the paddle and the contents were mixed in gentle circular motion in a horizontal plane. Astitis cases were determined according to the degree of agglutination. CMT is a highly sensitive and fast method to detect abnormally cell-rich milk. The principle con- sists of a mixture of milk and tea pools (detergent) in amounts intended to rupture the cells, so that the nuclear DNA of the cells gels when they come into contact with the latter [13]. Determination of somatic cell count Somatic cell count of the samples was determined via au- tomatic device. The device is a counter based on the optical fluorescence principle. Ethidium bromide enters the structure of nuclear DNA, producing a fluorescent signal corresponding to SHS in milk. It is advantageous because it is an automatic and fast technique. The milk sample was brought to room tempera- ture (21 ± 1°C) and homogenized. Then, 0.75 ml of Reagan C at room temperature was added to the clean 1.5 ml ependorfs and 0.75 ml of the milk sample was added. The ependorfs prepared for analysis were homogenized by vortexing. For somatic cell counting, a pre-prepared eppendorf sample at room temperature was drawn into the cassette and loaded into the instrument. After loading, the result is automatically saved by the device within 2 minutes. Bacterial isolation and identification Samples were brought to the laboratory by a cold chain and collected with sterile gauze from milk-filled tubes that were strea- ked in blood agar. The agar plates were incubated at 37 °C for 24-48 h under aerobic conditions. Gram staining is performed on colonies that have grown as a result of incubation. Gram- positive and Gram-negative samples were inoculated onto Tryp- tic soy agar. TSA Petri dishes were incubated for 24 h at 37 °C. Biochemical assays of pure colonies formed in TSA after incu- bation were verified using the BD Phoneix™ M50 device. This method is used for rapid identification and susceptibility testing of many aerobic and facultative anaerobic Gram positive and negative bacteria containing selected antimicrobial agents. 24- hour fresh cultures purified on triptic soy agar were suspended with ID broth in glass tubes according to McFarland 0.5 colony density. BD Phoneix ™ PMIC / ID87 was used for gram posi- tive bacterial isolates and BD Phoneix™ NMIC / ID-400 panel kit was used for gram negative bacterial isolates. Identification was performed on the instrument using separate panels for each sample. ID Broth suspension tubes prepared for each sample were placed in the device for bacterial diagnosis. Biochemical identification data obtained from the device were evaluated. Determination of antibiotic susceptibility Antibiotic susceptibility tests were performed for the isolates that were identified with the automated device using the BD Phoneix™ PMIC / ID87 and NMIC / ID-400 kit. 24 h fresh cul- tures purified on triptic soy agar were prepared with AST Broth present in glass tubes according to McFarland 0.5 colony den- sity. Panels containing bacterial suspensions were placed in the Highlights in BioScience Page 2 of 7 October 2021|Volume 4 http://bioscience.highlightsin.org/ Kirkan et al., 2021 Identification of pathogen bacteria from camel (Camelus dromedarius) mastitis device and MIC detection and sensitivity results were obtained from the electronic system. The antimicrobials and their MIC values of bacterial isolates (n=32) were determined according to CLSI standards [14]. Results California mastitis test and somatic cell count numbers A plastic shovel with four glasses was used. An equal amount (3 ml) of milk and reagent was placed in each glass of the shovel and the contents mixed in a horizontal plane with a slight circu- lar motion. According to the degree of agglutination, the CMT score was determined as "Negative" (-), suspicious "+/-", Mild (+), Significant (++) and Severe (+++) [9]. The CMT score and somatic cell numbers of 40 milk samples determined by the Nu- cleoCounter™ SCC-100 (Chemometec®, Denmark) device are shown in Table 1. Table 1. CMT Score and Somatic cell counts of milk samples. Milk samples (n=40) Milk samples CMT scores Somatic cell count standards 8 Negative (-) 100 16 Suspicious (+/-) 100-300 14 Mild (+) 300-900 2 Severe (++) 900-2700 Identification Bacterial growth was not observed in 8 samples taken from 4 (20%) female camels. Bacterial growth was detected in 16 (80%) female camel milk samples (n = 32 samples). As a result of identification, 4 (12.5%) Staphylococcus aureus, 4 (12.5%) Staphylococcus auricularis, 2 (6.25%) Staphylococcus pettenko- feri, 2 (6.25%) Staphylococcus cohnii spp. cohnii, 2 (6.25%) Staphylococcus equorum, 2 (6.25%) Staphylococcus capitis, 2 (6.25%) Streptococcus agalactiae, 2 (6.25%) Streptococcus dys- galactiae, 4 (12.5%) Escherichia coli, 2 (%) 6.25) Pseudomonas pseudalcaligenes, 2 (6.25%) Corynebacterium pseudotuberculo- sis, 2 (6.25%) Aerococcus viridans and 2 (6.25%) Gemella mor- billorum were identified (Table 2). Antibiotic susceptibility The evaluation of MIC values indicating the antibiotic sus- ceptibility of the strains identified in our study (n = 32) showed that S. aureus (n=4) strains were 100% resistant to Clindamycin and Trimethoprim-Sulfomethoxazole, and 75% resistant to Ery- thromycin, Penicillin G and Vancomycin. S. auricularis (n=4) strains were resistant to Daptomycin, Fusidic acid at a rate of 100%, and against Quinupristine dalfopristin and Trimethoprim- Sulfomethoxazole at a rate of 75%. S. pettenkoferi (n=2) strains were 100% resistant to Oxacillin and Penicillin G. S. cohniispp. cohnii (n=2) strains were 100% resistant to Ampicillin, Clin- damycin, Fosfomycin, Oxacillin, Penicillin G and Vancomycin.S. equorum (n=2) strains were 100% resistant to Ampicillin, Fu- sidic acid and Penicillin G. S. capitis (n=2) strains were exam- ined, it was found that the strains were 100% resistant to Van- comycin, Amoxicillin-Clavulanic acid and Ampicillin.S. agalac- tiae (n=2) strains were 100% resistant to Ciprofloxacin, Ery- thromycin, Fusidic acid, Gentamicin, Penicillin G and Teicop- lanin. S. dysgalactiae (n=2) strains were 100% resistant to Ampi- cillin, Cefoxacin, Ciprofloxacin, Daptomycin, Erythromycin, Fu- sidic Acid, Gentamicin, Nitrofurantoin, Penicillin G, Quinupris- tine dalfopristin, Rifampin and Teicoplanin. C. pseudotuberculo- sis (n=2) strains were 100% resistant to Ampicillin, Ciprofloxacin, Fusidic acid, Nitrofurantoin and Penicillin G. A. viridans (n=2) strains were 100% resistant to Ciprofloxacin, Erythromycin and Gentamicin. G. morbillorum (n=2) strains were 100% resistant to Ciprofloxacin, Erythromycin and Gentamicin. E. coli (n=4) strains were 100% resistant to Cefotaxime, Ciprofloxacin, Gen- tamicin, Meropenem, Netilmicin, Tigecycline and Trimethoprim- Sulfomethoxazole. P. pseudalcaligenes (n=2) strains were 100% resistant to Amoxicillin-Clavulanic acid, Aztreonam, Cefurox- ime and Trimethoprim-Sulfomethoxazole (Table 3) Discussion Subclinical camel mastitis has not been extensively studied and reported in prevalence studies. Subclinical mastitis, where macroscopic clinical symptoms cannot be seen and indirect tools are needed for diagnosis, remains a significant public health problem for camel populations and consumers. Consuming camel milk has effects on animal health and reduces the quantity and quality of milk. In studies on the relationship between the Cal- ifornia mastitis test (CMT) and the number of bacteria in the camel milk test, it was shown that a high proportion of subclin- ical mastitis was CMT positive and that there was a significant difference between positive / negative CMT cases [15-17]. In an- other study conducted in Sudan, CMT, somatic cell count (SCC) and bacterial isolates were compared and showed that the mean values of CMT and SCC were higher in samples taken from an infected mammary gland [18]. In addition, 47.3% of 336 milk samples tested positive in CMT and somatic cell count of 757 samples were reported to have a count range of 5 X 105 to 7.5 X 106 cells / ml [19]. CMT has a sensitivity of about 70% and a specificity of 91% in camel mastitis [20]. Additionally, in another study, it was suggested that detection of SCC in camels was more sensitive in detecting subclinical mastitis in camels than the N-acetyl beta-D- glucosaminidase test [21]. The prevalence of subclinical masti- tis in camels in Riyadh, Saudi Arabia was 33% in milk samples examined according to CMT [15]. In other studies, the over- all prevalence of mastitis was 44.8% and the prevalence of sub- clinical mastitis was 46% [16,17]. In this study, the CMT test and digital CCS were used to investigate subclinical mastitis in female camels. Milk samples were obtained from 20 camels for the diagnosis of subclinical mastitis and the CMT score was negative, contaminated with +1, +2 and +3. The mean CCS obtained from the assessed healthy camel was 100,000 cells/ml. In our study, no bacterial growth was found in animals with so- Highlights in BioScience Page 3 of 7 October 2021|Volume 4 http://bioscience.highlightsin.org/ Kirkan et al., 2021 Identification of pathogen bacteria from camel (Camelus dromedarius) mastitis Table 2. The distribution of bacteria isolated from samples. Milk samples with bacterial growth Bacrterial Identification Right lobe Left lobe Sample 1 Staphylococcus aureus Staphylococcus aureus Sample 2 Staphylococcus aureus Escherichia coli Sample 3 Staphylococcus auricularis Staphylococcus auricularis Sample 4 Staphylococcus aureus Staphylococcus auricularis Sample 5 Escherichia coli Staphylococcus auricularis Sample 6 Staphylococcus pettenkoferi Staphylococcus pettenkoferi Sample 7 Staphylococcus cohnii spp. cohnii Staphylococcus cohnii spp. cohnii Sample 8 Staphylococcus equorum Staphylococcus equorum Sample 9 Staphylococcus capitis Staphylococcus capitis Sample 10 Streptococcus agalactiae Streptococcus agalactiae Sample 11 Streptococcus dysgalactiae Streptococcus dysgalactiae Sample 12 Escherichia coli Escherichia coli Sample 13 Pseudomonas pseudalcaligenes Pseudomonas pseudalcaligenes Sample 14 Corynebacterium pseudotuberculosis Corynebacterium pseudotuberculosis Sample 15 Aerococcus viridans Aerococcus viridans Sample 16 Gemella morbillorum Gemella morbillorum matic cell counts less than 100,000 cells/ml. In our study, the prevalence of subclinical mastitis was determined to be 80% in milk samples randomly selected from healthy-looking camels. However, previous studies have shown that CMT has a sensitiv- ity of 70% and specificity of 91% in camel mastitis and there is a positive correlation between SCC/CMT scores [20-22]. Because early and effective treatment of mastitis is very important, accu- rate diagnosis of subclinical mastitis is always a priority. The technique in which the intranuclear dye penetrates into the DNA of the cell and the stained cells are detected by spectrophotome- ter has been shown to be applicable in the diagnosis of subclin- ical mastitis in camels. This study was confirmed by isolating and identifying the causative agent from milk samples from ani- mals evaluated for subclinical mastitis by determining CMT and SCC. Epidemiological data collected in recent years show that the prevalence of clinical mastitis in camels is between 31-38% in Europe and 31% in Uruguay. While the prevalence of general mastitis in camels was 18.52%, subclinical mastitis was more common (24.7%) than clinical mastitis (11.67%). The major pathogens isolated were Staphylococcus sp. (41.67%) followed by Streptococcus sp. (21.67%), Enterobacter sp. (15.00%), Corynebacterium pyogenes (10.00%), Micrococcus sp. (5.00%), Pasteurella sp. (5.00%) and Pseudomonas sp. 1.66%. In North Kordofan, Sudan, in 2013, the incidence rate was found to be 25% subclinical mastitis, 13.3% and 15% using other techniques such as SCC and White Side Test. The isolated pathogens were Staphylococcus sp. (80.30%), Bacillus sp. (9.09%), Pasteurella sp. (6.06%), Corynebacterium sp. (3.03%) and Streptococcus sp. (1.52%) [23]. Staphylococcus sp. As a result of the study conducted on 90 female camels in Jordan to identify and gen- erate data on camel mastitis and pathogens, 21% of the camels had clinical symptoms of mastitis and Micrococcus sp., Staphy- lococcus aureus, Streptococcus sp. and Corynebacterium sp. identified. The isolates were susceptible to antibiotics such as Gentamicin, Ampicillin and Tetracycline. The study concluded that Gram-positive cocci are predominantly the result of masti- tis [24]. The data obtained are in parallel with the findings of our research. In this study, 81% of Gram-positive bacteria were isolated. Staphylococcus sp., Streptococcus sp. and Micrococ- cus sp. isolated camels have been reported to be the dominant pathogen of these species [25,26]. The incidence and causes of mastitis in camels differ sig- nificantly due to geographical region and individual herd man- agement [17]. The prevalence of camel mastitis in Sudan was 30.2%, of which 4.9% were clinically and 25.3% subclinically in the Jigjiga region. There was a high rate of chronic form (72.41%) followed by acute form (24.14%), and three types of clinical mastitis were diagnosed, the least seen gangrenous form (3.45%). Clinical mastitis occurred in animals older than 10 years of age and late in lactation (55%). In our study, subclinical mastitis cases were detected in animals younger than 10 years of age. The dominant isolated microorganisms are Staphylococcus sp. (37.8%), E. coli (18.9%), Streptococcus sp. (13.5%), Bacil- lus sp. (10.8%), Micrococcus sp. (8.1%), Corynebacterium sp. (5.4%) and Salmonella sp. (5.4%) [27]. Of the mastitis (76.0%) evaluated in Eastern Ethiopia and observed in camel herds, Staphylococcus aureus (4.2%), Coagulase Negative Staphy- lococci (39.6%), Streptococcus agalactiae (3.5%), Streptococ- cus dysagalactiae (22.2%), Corynebacterium sp. (9%), Bacil- lus sp. (7.6%), Streptococcus uberis (7.6%), Escherichia coli (6.37%) were identified [28]. Similar to these findings, Staphy- lococcus sp., Streptococcus sp., E. coli, Corynebacterium pseu- dotuberculosis and Pseudomonas pseudalcaligenes species were identified in addition to A. viridans and Gemella morbillorum. Highlights in BioScience Page 4 of 7 October 2021|Volume 4 http://bioscience.highlightsin.org/ Kirkan et al., 2021 Identification of pathogen bacteria from camel (Camelus dromedarius) mastitis Table 3. Antibacterial susceptibility results of identified bacterial strains according to MIC values. AMC: Amoxicillin-Clavulanic acid, AM: Ampicillin, FOX: Cefoxitin, CIP: Ciprofloxacin, CC: Clindamycin, DAP: Daptomycin, E: Erythromycin, FF: Fosfomycin, FA: Fusidic acid, GM: Gentamycin, LVX: Levofloxacin, LZD:Linezolid, FM:Nitrofurantoin, OX: Oxacillin, P: Penicillin G, SYN: Quinupristin-dalfopristin, RA: Rifampin, TEC: Teicoplanin, TE: Tetracycline, NN: Tobramycin, SXT: Trimethoprim-Sulfomethoxazole, VA:Vancomycin, AN:Amikacin, ATM: Aztreonam, FEP: Cefepime, CAZ: Ceftazidime, CRO: Ceftriax- one, CXM: Cefuroxime, CL:Colistin, ETP:Ertapenem, IPM:Imipenem, MEM: Meropenem, NET:Netilmicin, PIP:Piperacillin, TZP: Piperacillin-Tazobactam, TGC:Tigecycline. AMC AM FOX CIP CC DAP E FF FA GM LVX LZD S I R S I R S I R S I R S I R S I R S I R S I R S I R S I R S I R S I R Staph. aureus (n=4) 2 1 1 1 1 2 2 2 1 2 1 4 4 1 3 1 3 2 2 2 2 3 1 4 Staph. auricularis (n=4) 3 1 4 4 3 1 3 1 4 3 1 4 4 4 3 1 2 2 Staph. pettenkoferi (n=2) 2 2 2 2 2 2 2 2 2 2 2 2 Staph. cohnii spp. cohnii (n=2) 2 2 1 1 2 2 2 2 2 2 2 2 2 Staph. equorum (n=2) 2 2 2 2 2 2 2 2 2 2 2 2 Staph. capitis (n=2) 2 2 2 2 2 2 2 2 2 2 2 Str. agalactiae (n=2) 2 2 2 2 2 2 2 2 2 2 2 2 Str. dysgalactiae (n=2) 1 1 2 2 2 2 2 2 2 2 2 2 2 C. pseudotuberculosis (n=2) 2 2 2 2 2 2 2 2 2 2 2 2 A. viridans (n=2) 2 2 2 2 2 2 2 2 2 2 2 2 G. morbillorum (n=2) 2 2 2 2 2 2 2 2 2 2 AN AMC AM ATM FEP CAZ CRO CXM CIP CL ETP GM S I R S I R S I R S I R S I R S I R S I R S I R S I R S I R S I R S I R E. coli (n=4) 3 1 1 3 4 4 4 4 4 4 4 2 2 3 1 4 P. pseudalcaligenes (n=2) 2 2 2 2 2 2 2 2 2 2 2 2 FM OX P SYN RA TEC TE TGC NN SXT VA S I R S I R S I R S I R S I R S I R S I R S I R S I R S I R S I R Staph. aureus (n=4) 4 2 2 1 3 3 1 4 3 1 4 4 4 4 1 3 Staph. auricularis (n=4) 3 1 2 1 1 3 1 1 3 3 1 4 4 4 4 1 3 4 Staph. pettenkoferi (n=2) 2 2 2 1 1 2 2 2 2 2 2 2 Staph. cohnii spp. cohnii (n=2) 2 2 2 2 2 2 2 2 2 2 2 Staph. equorum (n=2) 2 1 1 2 2 1 1 2 2 2 2 2 2 Staph. capitis (n=2) 2 2 2 2 2 2 2 2 2 2 2 Str. agalactiae (n=2) 2 2 2 2 2 2 2 Str. dysgalactiae (n=2) 2 2 2 2 2 2 2 C. pseudotuberculosis (n=2) 2 2 2 2 2 2 2 A. viridans (n=2) 2 2 2 2 2 2 2 G. morbillorum (n=2) 2 2 2 2 2 2 IPM MEM NET PIP TZP TGC SXT S I R S I R S I R S I R S I R S I R S I R E. coli (n=4) 1 3 4 4 4 4 4 4 P. pseudalcaligenes (n=2) 2 2 2 2 2 2 2 Besides, 23.8% of Gram-negative bacteria species have been reported in previous studies [29]. In our study, Gram negative species were determined as Pseudomonas pseudalcaligenes, es- pecially E. coli as a cause of circumferential mastitis. In our study, 12.5% of Staphylococcus aureus were isolated. S. auricu- laris, S. pettenkoferi, S. cohnii spp. cohnii, S. equorum, S. capitis were other Staphylococcal species isolated in this research. Our result (12.5%) was found to be lower than the results reported before [30] (44.82%). In the same study, E. coli (18.92%) was the second most isolated bacterium. In our study, E. coli was identified at a rate of 12.5%. Previous antimicrobial susceptibility tests showed high sus- ceptibility to commonly used antibiotics against bacteria. The isolates were found to be highly sensitive to Gentamicin , Cipr -ofloxacin, Cloxacillin and Amikacin, moderately sensitive to Ampicillin / Sulbactam and Trimoxazole, and highly resistant to Tetracycline and Chloramphenicol [16, 29, 30, 31, 32]. In the study conducted by Abdelgadir [25], Oxytetracycline, Tetracycline and Chloramphenicol were determined as effective antibiotics against mastitis pathogens in camels. According to the antibiotic susceptibility findings obtained in our study, Staphy- lococcus species were found to be susceptible to Daptomycin, Tetracycline, Levofloxacin, Tigecycline and Tetracycline antibi- otics and Trimetoprim-Sulfometoxazole and Penicillin G resis- tant. As an important result of antibiotic susceptibility find- ings, Vancomycin resistance in S. aureus and Staph. cohnii spp. Highlights in BioScience Page 5 of 7 October 2021|Volume 4 http://bioscience.highlightsin.org/ Kirkan et al., 2021 Identification of pathogen bacteria from camel (Camelus dromedarius) mastitis cohnii strains is remarkable. Streptococcus species identified in our study were found to be susceptible to Clindamycin, Lev- ofloxacin, Linezolid and Tetracycline and resistant to Penicillin G, Gentamicin, Erythromycin and Ciprofloxacin. Resistance to beta lactam group and macrolides was determined. Other Gram- positive strains identified as C. pseudotuberculosis, A. viridans and G. morbillorum were also susceptible to Amoxicillin-Clavu -lanic acid, Levofloxacin and Daptomycin, and resistant to Ampi- cillin and Penicillin G. Antimicrobial susceptibility of E. coli strains was found to be susceptible to Aztreonam, Cefepime and Piperacillin, and re- sistant to Gentamycin, Meropenem, Tigecycline, Trimethoprime -Sulfometoxazole and Amoxicillin Clavulanic acid; Pseudomonas pseudalcaligenes strains were susceptible to Cefepime, Ciproflox -acin, Piperacillin. Gram-positive bacteria identified in our study were found to be susceptible to Levofloxacin, Linezolid, Tetracycline, Dapto- mycin, and resistant to beta lactam group antibiotics and macro -lides, Vancomycin resistance was determined in S. aureus and S. cohnii spp. cohnii strains. Gram negative strains were generally susceptible to Cefepime and Piperacillin; resistant to Trimetho- prime -Sulphomethoxazole and Amoxicillin-Clavulanic acid. As a result, it has been shown that multiple antibiotic resistance de- velops in bacteria causing mastitis in camels. Our study suggests that subclinical mastitis in camels is more common than other forms of mastitis and that infected animals may be a source of contamination as a microorganism reser- voir; identification of both infectious and environmental masti- tis pathogens. Since the prevalence of mastitis in camels dif- fers considerably depending on geographical area and individ- ual herd management, it is recommended to use antibiotics to prevent the development of antimicrobial resistance as well as mastitis control methods; such as elimination of existing infec- tion, prevention of new infection and monitoring the health sta- tus of the mammary. Considering that camel milk is used raw for human consumption without heat treatment in order not to affect the nutritional and immunological factors it contains, the pathogens causing subclinical mastitis can easily threaten public health, and it is essential to inform the farmers and consumers about the related pathogens. Acknowledgement This research was funded by Aydın Adnan Menderes Univer- sity Scientific Committee by grant number VTF-18017. References 1. Korhonen H, Pihlanto A. Food-derived bioactive peptides oppor- tunities for designing future foods. Curr. Pharm. Des. 2001, 9: 1297-1308. 2. 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Cross-sectional study of mastitis in camels (Camelus dromedarius) in selected sites of Ethiopia[dissertation]. Freie Universität Berlin and Addis Ababa University; 2001. 26. Alamin MA, Alqurashi AM, Elsheikh AS, Yasin TE. Mastitis incidence and bacterial causative agents isolated from lactating she-camel (Camelus dromedarius). J. Agric. Vet. 2013, 2(3): 7-10. 27. Blood DC, Radostits OM. Veterinary Medicine: A Textbook of disease of cattle, sheep, pigs, Goats and Horses. Baillion Tindall: London; 2007. 28. Eyassu S, Bekele T. Prevalence and etiology of mastitis in tra- ditionally managed camels (Camelus dromedarius) in selected pastoral areas in eastern Ethiopia. Ethiop. Vet. J. 2010, 14(2): 103-113. 29. Al-Tofaily, Y.I., and Alrodhan, M.A.N.: Study on clinical mas- titis (Bacteriological) in she-camels (Camelus dromedaries) in some areas of middle Euphrates in Iraq. QJVMS. 2011, 10(2): 66-76. 30. Al-Juboori AA, Kamat NK, Sindhu JI. Prevalence of some masti- tis causes in dromedary camels in Abu Dhabi, United Arab Emi- rates. Iraqi J. Vet. Sci. 2013a, 27(1): 9-14. 31. Fazlani SA, Khan SA, Farazl S, Awan MS. Antimicrobial sus- ceptibility of bacterial species indentified from mastitic milk sam- ples of camel. Afr. J. Biotechnol. 2011, 10(15): 2959-2964. 32. Alqurashi AM, Alamin MA, Elsheikh AS, Yasin TE. Sensitivity of bacterial isolates from mastitic she-camel (Camelus dromedaries) to antibiotics. Am.J. Sci. 2013, 9(4): 47-52. Highlights in BioScience Page 7 of 7 October 2021|Volume 4 http://bioscience.highlightsin.org/ Abstract Introduction Materials and Methods Sample collection Ethics of animal use California mastitis test Determination of somatic cell count Bacterial isolation and identification Determination of antibiotic susceptibility Results California mastitis test and somatic cell count numbers Identification Antibiotic susceptibility Discussion Acknowledgement References