East East Afr. J. Biophys. Comput. Sci. (2023), Vol. 4, Issue. 2, 1-12 *Corresponding author: Email: msulayeman@gmail.com https://dx.doi.org/10.4314/eajbcs.v4i2.1S Sero-epidemiology of Foot and Mouth Disease and Farmers Perception on Vaccinating Cattle against the Disease in Sidama Region, Southern Ethiopia Mishamo Sulayeman*1, Tamirat Demissie1, Ayelech Muluneh2, Gizachew Hailegebreal1, Sultan Abda1 1Faculty of Veterinary Medicine, Hawassa University, P.O. Box 05, Hawassa, Ethiopia. 2National Animal Health Diagnostics and Investigation Center, P.O.Box 04, Sebeta, Ethiopia. KEYWORDS: Cattle; Ethiopia; Farmers perception; FMD; Sero-prevalence; Vaccination ABSTRACT Ft and mouth disease (FMD) is a severe, highly cntagious viral disease of livestck that has a significant economic impact. A cross-sectional study was conducted from September 2019 to June2020 in three selected districts of Sidama region, Southern Ethiopia with the objectives of determining the sero-prevalence of cattle against foot and mouth disease virus (FMDV), identifying potential risk factors and assessing farmers’ perception on vaccination against FMD. Purposive and systematic random sampling techniques were employed to select the districts and study animals, respectively. A total of 510 cattle were tested for FMDV antibodies using 3ABC-ELISA. The overall cattle and herd level sero-prevalence were 15.5% and 24.7%, respectively. Among the considered risk factrs, age of the animal, herd size and season were significantly associated with the sero-positivity of FMDV (P<0.05). Out of 120 farmers interviewed 84.2% had never vaccinated their cattle against FMDV. Inaccessibility (83.7%) and unaffordable cost (72.1%) of the vaccine were mentioned as leading causes for the low vaccination practice in the current study areas. Majority of the respondents (68.3%) don’t perceive vaccinating cattle against FMDV as one of the preventive measures. In districts with lower perception of farmers on vaccinating their cattle against FMDV, higher sero-prevalence of the disease were recorded. The present serological and questionnaire survey indicated that the presence of FMD sero-positive animals in the current study areas. Therefore, an integrated strategy for disease control has to be designed and implemented which could include enhancing farmers’ perception about use of vaccination in preventing FMD and government provision of vaccines at affordable cost to the farmers. . INTRODUCTION Ethiopia is one of the countries that possess a huge number of livestock populations in the Africa continent estimated to be 56.5 million cattle, 30.7 million sheep and 30.2 million goats were found in the country (CSA, 2017). The livestock sector contributes about 40% of the agricultural Gross Domestic Product (GDP) and nearly 20% of total GDP, and 20% of national East African Journal of Biophysical and Computational Sciences Journal homepage : https://journals.hu.edu.et/hu-journals/index.php/eajbcs Hawassa University College of Natural & Computational Sciences Year 2021 Volume xx No xx Research article mailto:msulayeman@gmail.com https://dx.doi.org/10.4314/eajbcs.v4i2.1S East Afr. J.Biophys.Comput. Sci. (2023), Vol. 4, No. 2, 1-12 2 foreign exchange earnings in 2017 (World Bank, 2017).Within the cattle population, FMD occurs endemically resulting in several outbreaks every year (Ayelet et al., 2012). The causative agent, FMD virus (FMDV), has seven recognized serotypes (O, A, C, SAT 1, SAT 2, SAT 3 and Asia 1), with discrete immunologic, antigenic and genetic properties. They also differ in distribution across the globe (FAO, 2007). Five of the seven serotypes of FMD (O, A, C, SAT 2, SAT 1) were identified in Ethiopia (Rufael et al., 2008; Ayelet et al., 2009; Negusssie et al., 2010). Serotype C was not identified after 1983; however, a serotype C- specific antibody in cattle was reported (Rufael et al., 2008). Morbidity has been reported to reach as high as 100% in susceptible animal populations but it is rarely fatal except in young animals (Kahn and Scottline, 2005). Infected animals show a spectrum of responses to FMD ranging from unapparent infection to severe disease and death (OIE, 2008). Foot and mouth disease is endemic with high prevalence in Africa, the Middle East, and Asia and is also present in parts of South America (Rweyemamu and Astudillo, 2002). The disease is endemic in Ethiopia and remains largely uncontrolled due to the absence of prophylactic vaccination except for a few dairy herds containing imported breeds (Sahle, 2004;Megersa et al., 2009). Serological surveys reported a sero-prevalence that ranges from 5% to 72.1% at the animal level in different parts of the country (Bayissa et al., 2011, Sulayeman et al., 2018; Shazali et al., 2021). In terms of livestock exports from Africa, FMD is often perceived as a major hindrance to international trade (Thomson et al., 2004). In part, this perception is based on the assumption that disease freedom is required before export is possible, and has resulted in costly and an elaborated FMD control measures such as disease-free zones in Southern Africa and elsewhere (Bruckner, 2004). Commodity based approaches can provide an acceptable level of risk for exported livestock or livestock products according to international standards(Thomson et al., 2004), but in the case of FMD, they still require an understanding of FMD status in cattle entering the market chain. Recommended control measures for FMD include animal movement restrictions, a vaccination programme, animal quarantine, environmental sanitary controls, outbreak investigation, serological surveillance and slaughtering of sick animals (Chaosuancharoen, 2012). However, it is a global problem since the result of the increasing movement of human and livestock and livestock products (Perry, 2007). This is mainly due to lack of vaccination, free livestock movement among different regions in the countries and across international borders, the existence of multiple FMD virus serotypes, and involvement of wildlife (Sahle, 2004; Rufael et al., 2008). Studies undertaken in Ethiopia revealed that the disease is still endemic and occurs in different parts of the country (Sulayeman et al., 2018; Shazali et al., 2021). There is neither a nationwide control strategy nor a legislation for making FMD notifiable to the veterinary authorities or for animal movement restrictions to be imposed. Therefore, livestock is at risk from endemic strains as well as from antigenic variants prevailing in neighboring countries (Sahle, 2004). There is a difference in the epidemiology and economic impacts of FMD in the livestock East Afr. J.Biophys.Comput. Sci. (2023), Vol. 4, No. 2, 1-12 3 production systems (Jemberu et al., 2014) in different parts of the country. Unidentified farmers’ perceptions about risk of the disease, lack of pragmatic vaccination schemes and presence of unrestricted animal movement regardless of certification are the major reasons that could intensify the distribution of FMD alongside the cattle market chain. Despite this fact, there is no published information regarding the status of FMD and farmers’ perceptions and practices on vaccinating their cattle against the disease in Sidama region. Therefore, this study was aimed to generate current information on the sero- prevalence status of FMD and predisposing risk factors and assesses farmers’ perception on vaccination against FMDV in selected districts of Sidama region, Southern Ethiopia. MATERIALS AND METHODS Description of the study area The study was carried out in three purposively selected districts of Sidama region, namely Hawassa zuria, Boricha and Wondo Genet. Sidama region is located northeast of Lake Abaya at an altitude of l500 to 2500 m.a.s.l. The region has gegraphic coordinates of latitude, north,5′ 45″to 6′ 45″ and lngitude, east, 38′ to 39′. Mean annual rainfall of this area varies between 1200 mm and 1599 mm, with 15°C- 19.9°C average annual temperature (CSA, 2015) (Figure 1). . Figure 1: Map showing the study areas East Afr. J.Biophys.Comput. Sci. (2023), Vol. 4, No. 2, 1-12 4 Study design and sampling strategy A cross-sectional study design was implemented for sero-prevalence study of antibodies against FMDV in the study areas. Hawassa Zuria, Boricha and Wondo Genet districts were selected purposively based on their transport accessibility, gegraphical location and presence of large cattle population. From each district 30% of kebeles (the smallest administrative units in Ethiopia) were selected using simple random sampling. From each kebeles, 20% of privately owned herds were randomly selected. From each herd, the study animals (cattle) were then selected using simple random sampling method to achieve the required sample size. The alleged potential risk factors for the occurrence of the disease such as age, breed, sex, districts, herd size and composition, season, vaccination history and management were also recorded. Sampled animals were categorized based on their breed (local and cross), sex (female and male), herd composition (cattle only and cattle mixed with small ruminants), vaccination history (previously vaccinated and non-vaccinated) and management types (intensive and semi- intensive). Ages (young, adult and old) were categorized based on their dental eruption status (Berecha et al., 2011) and herd sizes where also classified as large farms size, with more than 50 animals, medium (20 to 50 animals) and small (< 20 animals) (Edao et al., 2018). Study animal population Local and cross breeds of cattle kept under intensive and semi-intensive system were included. According to Pace and Wakeman (2003), the age groups of cattle were categorized as (≤3.5years) Young, (3.5years‐5.5years) Adult and (> 5.5years) Old. Sample size determination The sample size required for the study was calculated based on the following formula (Thrusfield, 2005). n = Z2*Pexp (1- Pexp) d2 Where, n= required sample size, Z= statistic for level of confidence = 1.96, Pexp = expected prevalence, 95% confidence level and d2 = absolute desired precision of 0.05. Accordingly, based on the above formula and 9.5% expected prevalence (Megersa et al., 2009), the sample size computed for animal level prevalence was 132. To increase the precision, calculated sample size was made four fold to 528, but due to shortage of sample collection materials only 510 cattle were considered. This number was allocated proportionally to the respective districts based on the total cattle population in each districts. Serum sample collection From each cattle, about l0 ml of blood sample was collected from the jugular vein and kept overnight on a table at room temperature. Then serum was aspirated with pasture pipette and transferred into cryovial and transported to Hawassa University Veterinary microbiology laboratory for storage at −20 °C. All the sera were transported with an ice box containing ice packs to the National animal health diagnostic and investigation center (NAHDIC) for serological test. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8464255/#vms3574-bib-0046 East Afr. J.Biophys.Comput. Sci. (2023), Vol. 4, No. 2, 1-12 5 Serological diagnostic tests Sera collected from bovine species was tested by FMDV 3ABC-Ab ELISA (ID Screen®) for the detection of antibody to poly protein called 3ABC which is a useful indicator of FMD virus infection regardless of the serotype involved (Haas, 1997; Mackay et al., 1998). Antibody to 3ABC (nonstructural protein) is found only in virus infected cattles but not in vaccinated animals (De Diego et al., 1997). Briefly, the test was carried out stepwise as per the manufacturer’s manual. First, all reagents were kept at room temperature and homogenized by vortex. The test was carried out in 96 well micro plates. Then 50μl of dilution buffer18 were added in to each well, 30μl of positive control were added in to wells A1 and B1, and the same volume of negative control were also added to wells C1 and D1, the rest wells were filled by 30μl of test sera. Then incubated at 37oC for 2 hours, after incubation the wells were emptied with washing 5 times with 300μl of wash solution along with paying great attention to avoid drying of wells between washing. After washing l00μl of the conjugate lX were added in to each wells and incubated for 30 min at 21oC. The wells were then emptied and washed 5 times with 300μl of wash solution, then l00μl of the substrate solution was added in to each wells and incubated at 2loC for l5 minutes in the dark. After adding a 100μl of Stop Solution to each well, the optical density (OD) reading was noted using a photometer at wavelength of 450 nm within 2 hours after the addition of the stop solution. Questionnaire survey Data concerning farmers’ perception towards vaccinating their cattle was collected by using a semi-structured pre-tested questionnaire. It was administered by interviewing individuals selected by systematic random sampling. Before the interview, the objective of the survey was properly explained and verbal consent was obtained from the respondents. The interviews were conducted in local languages (Sidaamu Afoo or Amharic). A total of 120 farmers, 40 farmers from each three districts were interviewed for the questionnaire survey. Data management and statistical analysis Data generated from the laboratory investigations and survey was recorded and coded using a Microsoft excel spread sheet (Microsoft Corporation) and analyzed using STATA version 13.0 for Windows (Stata Corp. College Station, TX, USA). The association between dependent and independent variables was analyzed at individual cattle level by using univariable and multivariable lgistic regression. Multivariable logistic model was used for variables with a p-value ≤ 0.05 on univariable analysis model. Further selection of variables in the final model was based on stepwise backward elimination procedure. Accordingly, Odds ratio (OR) was used to assess the strength of association between the putative risk factors and sero-positivity of the disease. RESULTS FMD sero-prevalence and risk factors Based on the total 510 sampled cattle, overall sero-prevalence of FMD was 15.5% and 24.7% at the individual animals and herd levels, respectively. Comparatively higher sero- prevalence (32.4%) was recorded in Hawassa East Afr. J.Biophys.Comput. Sci. (2023), Vol. 4, No. 2, 1-12 6 zuria district (p=0.02; OR=2.67%; 95%CI=1.14- 5.33) (Table 1). Table 1: Individual cattle level and herd level sero-prevalence of FMD Districts Farmers associations Individual cattle Herds Tested Positive (%) Tested Positive (%) HawassaZuria Labu-koromo 68 9(13.2) 27 8(29.6) Udo-wotate 65 16(24.6) 22 7(31.8) Galo-argisa 64 12(18.7) 22 8(36.4) Sub total 197 37(18.8) 71 23(32.4) Boricha Konser-fulasa 50 16(32) 31 11(35.5) Fulasa-aldada 48 5(10.4) 24 5(20.8) Hanja-chefa 52 7(13.5) 36 7(19.4) Aldada-dela 58 5(8.6) 27 3(11.1) Sub total 208 33(15.8) 118 26(22) Wondo Genet Watara-qachama 56 5(8.9) 13 2(15.4) Abayye 49 4(8.2) 20 4(20) Sub total 105 9(8.6) 33 6(18.2) Total 510 79(15.5) 222 55(24.7) Risk factors Breed of the cattle, sex, age, districts, herd composition and size, season, vaccination history and management types were the major exposure or predictor variables considered to predict the response of the outcome variable. Table 2:Lgistic regression analysis of FMD and its putative risk factors for sero-positivity of cattle. Risk factors Category No examined Prevalence Univariable Multivariable No (%) positive OR 95% CI P- value OR 95% CI P- value Districts Wondo genet 105 9(8.6) Ref. - - - - - Hawassa zuria 197 37(18.8) 2.67 1.14 – 5.33 0.02 0.63 0.37 – 5.23 0.63 Boricha 208 33(15.8) 2.01 0.92 – 4.37 0.04 1.38 0.16 – 2.48 0.51 Age Young 58 3(5.2) Ref. - - - - - Adult 67 5(7.5) 1.47 0.33 – 6.47 0.03 1.38 0.31 – 6.23 0.67 Old 385 71(18.4) 4.14 1.26 – 13.6 0.02 3.60 1.04 – 12. 47 0.04 Herd size Small 262 31(11.8) Ref. - - - - - Medium 229 46(20.1) 1.87 0.14 – 3.07 0.01 2.18 1.27 – 3.76 0.005 Large 19 2(10.5) 0.87 0.19 – 3.97 0.46 1.03 0.21 – 5.03 0.97 Managt. type Intensive 78 6(7.7) Ref. - - - - - Semi- intensive 432 73(16.9) 2.44 1.02 – 5.82 0.04 2.65 0.34 – 20.97 0.35 Season Wet 38 14(36.8) Ref. - - - - - Dry 472 65(13.7) 0.27 0.13 – 0.56 0.000 0.18 0.07 – 0.42 0.000 OR= odds ratio; CI= confidence interval Most of the documented variables revealed a high degree of association with FMDV infection or sero-positivity. The final multivariable lgistic regression model (Table 2) revealed that age, herd size and season were significantly associated with the sero-prevalence of the disease (P<0.05). Old cattle were 3.6 times at a higher risk of FMD than young cattle. East Afr. J.Biophys.Comput. Sci. (2023), Vol. 4, No. 2, 1-12 7 Farmers’ perception and practices related to FMD Out of 120 respondents 64(53.3%) and 56 (46.7%) of them responded that their production is dairy cattle and mixed production type (Table 3). Bovine pasteurellosis, blackleg, lumpy skin disease, anthrax and FMD were listed in order of vaccination practice by the respondents (Figure 2). Figure 2: List of diseases and farmers practice to vaccinate their cattle against different animal disease. Among the major cause for low vaccination practices against FMD in the study area, inaccessibility and unaffordable cost of the vaccine were mentioned by 83.7% (36/43) and 72.1% of the respondents, respectively. Moreover, 68.3% (82/120) of the farmers interviewed don’t perceive vaccination as a preventive measure for the disease (Table 4). Table 4: Farmer’s perception on vaccinating their cattle Variables Response Frequency (%) Dairy cattle production type Yes 64(53.3) Mixed production type Yes 56(46.7) Vaccinated their cattle against disease Yes 120(100) Perceive as vaccination is better than treatment Yes 104(86.7) Perceive vaccination as preventive measure against FMD Yes 38 (31.7) Vaccinated their cattle against FMD Yes 19(15.8) FMD is a common disease Yes 77(64.2) Pervious occurrence of FMD in the farm Yes 66(55%) Information about FMD Yes 108(90) Information about FMD vaccination Yes 43(35.8) 0.00% 10.00% 20.00% 30.00% 40.00% 50.00% 60.00% 70.00% 80.00% 90.00% 100.00% Bovine pasteurellosis Black leg disease Lumpy skin disease Anthrax disease FMD 94.20% 90.00% 84% 60.80% 15.80% V ac ci n at in g a n im al s ag ai n st t h e d is ea se s East Afr. J.Biophys.Comput. Sci. (2023), Vol. 4, No. 2, 1-12 8 Sero-prevalence study of FMD Overall sero-prevalence of FMD recorded in this study (15.5%)was in agreement with the previous findings of15.4% (Mohamoud et al., 2011) and 14.05% (Zerabruk et al., 2014)from Jijiga zone and Tigray respectively. In contrast, it is higher than previous reports made from different parts of Ethiopia which range from 4.8% - 12.08% (Negussie et al., 2011; Abunna et al., 2013; Beyene et al., 2015; Gelana et al., 2016; Belina et al., 2016). Compared to the present finding relatively higher sero-prevalence in bovine was reported as, 24.22%, 38.9% and 21.4% from central Ethiopia (Sulayeman et al., 2018), Borena (Melkamsew, 2018) and West Ethiopia (Desissa et al., 2014) respectively. Similarly, higher sero-prevalence of the disease was also reported from the neighboring countries of Africa, 52.5% in Kenya (Kibore et al., 2013), 61% in Uganda (Miaron et al., 2004) and 72.62% in Nigeria (Lazarus et al., 2012). These differences in the prevalence of the disease among the studies could be partly explained by the variation in agro-ecology; epidemiology of the disease and variations in the production or herding systems, vaccination coverage against FMD vaccine, immune status, interaction with cattle with other animals like small ruminants and management type of different study areas. Cattle managed semi-intensively were shown higher sero-prevalence than those kept under intensive management. Similarly, higher sero- prevalence was previously recorded in cattle kept under semi-intensive managements (Bedru, 2006). Free movement of animals for watering point and grazing areas, and relatively larger herd holding capacity were the possible causes for the disease prevalence difference in different management system. This is supported by the work of previous studies report that the movement of animals in search of feeds from one area to another and interaction of small ruminants is a significant risk factor for the occurrence of FMD (Gelaye et al., 2005;Fevre et al., 2006; Habiela et al., 2010). In the current study, sero-prevalence of FMD was significantly higher in old animals than in young groups. Similar findings were also previously reported from central Ethiopia (Sulayeman et al., 2018) and Awbere and Babille districts of Jijiga zone (Mohamoud et al., 2011). Aged caatle are more probably to have been exposed to FMDV during their lifetime and have developed immunity to the virus. Additionally, old animals are driven freely in grazing and watering points where infection could increase by contact (Jenbere et al., 2011). Higher sero-prevalence was recorded during the dry seasons, which might be associated with herd movement to grazing area after crops were collected. This finding is supported by previous study as dry season increase the risk of FMD occurrence (Sarker et al., 2011)and also described as FMD is a seasonal disease mostly seen during the dry season (Jibat et al., 2013). Because during the dry season, cattle may experience physiological stress due to the factors such as high temperatures, low humidity, and limited availability of fresh forage and water. This can deteriorate their immune response, making them more susceptible to FMD infection and increasing the sero-prevalence. Nearly 87% of the respondents’ perceived vaccination is better than treatment, but only East Afr. J.Biophys.Comput. Sci. (2023), Vol. 4, No. 2, 1-12 9 15.8% of them had vaccinated animals against the disease. Similarly Megersa et al (2009) reported that vaccine as prophylactic measures against FMD was accepted by most farmers, but very few of them regularly vaccinate their animals. On other hand, some farmers did not consider vaccination of FMD as significant prevention methods due to self-limiting disease and low mortality among affected animals. In a district, Wondo genet, where farmers perceive and practice vaccine as a preventive measure, lower FMD sero-prevalence was recorded than the other districts. The study further revealed that 90% of respondents had information about FMD in the selected districts. Similarly 92.5% awareness level was also previously reported from Bale zone (Misgana et al., 2013). From the respondents only15.8% vaccinate their cattle against FMD. Lower vaccination practice against FMD was also reported from Nigeria (Olabode et al., 2014). However, comparable higher vaccination practices against the disease were reported from Tanzania (Miaron et al., 2004, Moenga et al., 2013). Inaccessibility and unaffordable cost of the vaccine were mentioned as a leading cause for the low vaccination practice in the current study areas. Moenga et al. (2013) and Soko et al. (2018) were also stated that aforementioned causes were the major reason for lower vaccination practices of the farmers. Most of the respondents from the selected districts had experienced FMD outbreak in their farm at least once before the interview. In line with this investigation previous work reported that FMD is endemic, widely distributed and frequently noted in different farming systems and agro-ecological zones of the country (Asfaw and Sintaro, 2000; Sahle, 2004; Leforban, 2005). Despite this fact 82(68.3%) of the farmers interviewed don’t perceive vaccination as preventive measure for the disease. In line with this finding most livestock owners don’t perceive vaccinating animals against FMDas one of the important preventive measures (Moenga et al., 2013; Soko et al., 2018). CONCLUSIONS & RECOMMENDATIONS The present serological study indicates that the presence of FMD sero-positive animals in the current study areas. Semi-structured questionnaire based surveys indicated that farmers’ awareness about FMD vaccine is very low. Even some farmers’ having awareness on FMD vaccine, their perception on vaccinating cattle against FMD is significantly low due to inaccessibility and unaffordability of the vaccine. The current research work explored the complex epidemiological situation of FMD and farmer’s perception on vaccine against the disease; thus needs more detailed investigation for vaccine- based control methods and improved veterinary extension services. Authors’ contribution All authors included in this article are directly or indirectly participated in the planning, execution & analysis of this study. MS and TD participated in data gathering, statistical analysis and writing up of the final manuscript. GH andSA participated in editing of the manuscript. AM assisted the laboratory test. All authors read and accepted the final manuscript. Funding This research work was funded by Hawassa University. East Afr. J.Biophys.Comput. Sci. (2023), Vol. 4, No. 2, 1-12 10 Ethics approval and consent to participate The study was approved by college of natural and computational science research proposal review committee, Hawassa University. Oral informed consent was obtained for both questionnaires interview and blood sample collection to keep the privacy of specific farmers at the time of sample collection. All methods employed for this research were carried out in accordance with pertinent guidelines and regulations. Competing interests Authors declare no conflict of interest. Acknowledgements The authors extend gratitude to Hawassa University and NAHDIC for financial support and laboratory logistics provision. The authors are also thankful to the three district veterinary administrative farm owners and respondents who participated in this study. References Abunna F., Fikru S. and Rufael T. 2013. Sero-prevalence, of FMD at Dire Dawa and its surroundings, Eastern Ethiopia. Glob. 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