The variable prevalence of bovine tuberculosis among dairy herds in Central Ethiopia provides opportunities for targeted intervention.

Bovine tuberculosis (bTB) is an important disease for dairy productivity, as well as having the potential for zoonotic transmission. Previous prevalence studies of bTB in the dairy sector in central Ethiopia have suggested high prevalence, however, they have been limited to relatively small scale surveys, raising concerns about their representativeness. Here we carried out a cross sectional one-stage cluster sampling survey taking the dairy herd as a cluster to estimate the prevalence of bTB in dairy farms in six areas of central Ethiopia. The survey, which to date is by far the largest in the area in terms of the number of dairy farms, study areas and risk factors explored, took place from March 2016 to May 2017. This study combined tuberculin skin testing and the collection of additional herd and animal level data by questionnaire to identify potential risk factors contributing to bTB transmission. We applied the single intradermal cervical comparative tuberculin (SICCT) test using >4mm cut-off for considering an individual animal as positive for bTB; at least one reactor animal was required for a herd to be considered bTB positive. Two hundred ninety-nine dairy herds in the six study areas were randomly selected, from which 5,675 cattle were tested. The overall prevalence of bTB after standardisation for herd-size in the population was 54.4% (95% CI 48.7-60%) at the herd level, and it was 24.5% (95% CI 23.3-25.8) at the individual animal level. A Generalized Linear Mixed Model (GLMM) with herd and area as random effect was used to explore risk factors association with bTB status. We found that herd size, age, bTB history at farm, and breed were significant risk factors for animals to be SICCT positive. Animals from large herds had 8.3 times the odds of being tuberculin reactor (OR: 8.3, p-value:0.008) as compared to animals from small herds. The effect of age was strongest for animals 8-10 years of age (the oldest category) having 8.9 times the odds of being tuberculin reactors (OR: 8.9, p-value:<0.001) compared to the youngest category. The other identified significant risk factors were bTB history at farm (OR: 5.2, p-value:0.003) and cattle breed (OR: 2.5, p-value: 0.032). Our study demonstrates a high prevalence of bTB in central Ethiopia but with a large variation in within-herd prevalence between herds, findings that lays an important foundation for the future development of control strategies

Isolation, molecular characterization and sero-prevalence study of foot-and-mouth disease virus circulating in central Ethiopia

Abstract Background Ethiopian livestock production and productivity is still very low due to widespread of diseases. Among the diseases, foot-and-mouth disease (FMD) is an extremely contagious and acute viral disease that causes significant economic problems in the country. A cross sectional study design was conducted from September 2015 to May 2016 to isolate and characterize FMD virus from outbreak cases; determine the sero-prevalence of antibodies against FMD virus (FMDV), and assess potential risk factors associated with sero-prevalence of the disease in selected areas of central Ethiopia. A multistage sampling technique was employed to select the study animals. Isolated viruses were characterized by antigen ELISA (IZLER, Brescia, Italy) and by genetic analysis of the sequence of the viral protein 1 (VP1). Sero-prevalence was determined using an ELISA for antibodies against non-structural proteins of FMDV based on the 3ABC proteins (ID Screen® FMD NSP Competition, ID-VET, Grabels, France). Risk factors for sero-prevalence of antibodies against FMD virus was investigated using logistic regression analysis. Result From outbreak investigation, 28.8% (n = 378) cattle showed signs and lesions suggestive of FMD and 34 samples were subjected to virus isolation. Twenty eight of these cultures exhibited cytopathic effect (CPE) and were serotyped as O, A and SAT 2 FMD viruses. One A and two SAT 2 isolates named A-ETH-19-2015, SAT 2-ETH-18-2015 and SAT 2-ETH-20-2015 were further characterized by phylogenetic analysis. The overall sero-prevalence of antibodies against non-structural proteins of FMDV was 24.2% (n = 574). Cattle herds with crossbreed cattle, with older cattle (> 2 years), and kept together with small ruminants had higher sero-prevalences of antibodies against FMDV (p < 0.05). Conclusion This study showed that FMD was present in the study areas. Among the associated risk factors, breed, age and herd composition were significantly associated with presence of antibodies against FMD virus. Three different serotypes (A, O and SAT 2) were responsible for the outbreaks of the disease. Genetic analysis indicated that the isolated viruses clustered differently from previous outbreaks. Thus, further molecular analyses coupled with protection potential of the existing vaccines against the isolates should be performed

Evaluation of Postmortem Inspection Procedures to Diagnose Bovine Tuberculosis at Debre Birhan Municipal Abattoir

Routine meat inspection in the abattoir was used to examine carcass for subsequent approval for consumption. However, the chance of missing lesions results in approval of carcass and/or the offal with lesions of tuberculosis. A cross-sectional study was conducted at Debre Birhan Municipal abattoir from October 2016 to May 2017. Lesion prevalence estimation and two meat inspection procedures’ efficacy evaluation was attempted. The breeds of the animals inspected were zebu breeds. Routine abattoir meat inspection involves visual inspection, palpation and incision of intact organs such as the liver and kidneys, as well as inspection, palpation and incision of tracheobronchial, mediastinal and prescapular lymph nodes. The detailed meat inspection involves inspection of each of the carcass. In this case, the seven lobes of the two lungs, lymph nodes and organs were also thoroughly examined. The cut surfaces were examined under bright light sources for the presence of an abscess, cheesy mass, and tubercles in detail. The study involved and compared both routine and detailed meat inspections at the abattoir. Chi-square test of independence and odds ratio were used to see the association of lesion and different risk factors. Based on detailed meat inspection, the overall lesion prevalence of bovine tuberculosis in the carcass of cattle slaughtered at Debre Birhan municipality abattoir was found to be 4.7% but only 0.5% of the carcass examined had detectable bovine tuberculosis lesions when routine abattoir meat inspection alone was used. The majority of the lesions were distributed to the lungs and associated lymph nodes. There was a significant association (p &lt; 0.05) in TB infection rate and body condition score. In conclusion, this study has clearly indicated the prevalence of bovine tuberculosis lesions in the abattoir that are missed by routine abattoir meat inspection. In addition, it showed low sensitivity of the routine meat inspection procedure used. Hence, our study warrants immediate attention to strengthen the current meat inspection practices at Debre Birhan public abattoir

Seroprevalence and risk factors of lumpy skin disease in Ethiopia

Lumpy skin disease (LSD) is an acute or inapparent viral disease of cattle which is endemic in many African and Middle East countries. LSD is one of the major transboundary livestock diseases in Ethiopia. A cross-sectional study using multistage cluster sampling was undertaken in central and north-western parts of Ethiopia with the objectives to estimate seroprevalence and to identify and quantify risk factors contributing to the occurrence of the disease. A total of 2386 cattle sera were sampled from 605 herds and 30 clusters (kebeles) located in 10 districts and tested for presence of LSD virus antibodies using virus neutralization test. All the serum samples were collected from cattle having no history of LSD vaccination. The overall animal level and herd level apparent seroprevalences were 25.4% (95% CI: 23.7–27.2) and 48.9% (95% CI: 44.9-52.9), respectively and varied significantly between districts. The true animal level and herd level prevalences were estimated as 26.5% (95% CI: 24.7–28.3) and 52.6% (95% CI: 48.3–56.9), respectively. At animal level, adult age (OR = 2.44 (95% CI: 1.67–3.55) compared to calf), contact with other animals (OR = 0.41 (95% CI: 0.23-0.74), compared to no contact) and presence of water bodies (OR = 1.61 (95% CI: 1.03–2.52), compared to no such bodies) were identified as the most important risk factors in relation to testing LSD positive. The putative risk factors altitude, breed, sex, and presence of animal trade route showed no significant association with LSD sero-status. Generally, cattle population with many adult animals and that live in wet areas are at highest risk, whereas cattle in frequent contact with other animals and animal species have lower risk, potentially due to a dilution effect of vectors.</p

Temporal and Spatial Patterns and a Space&ndash;Time Cluster Analysis of Foot-and-Mouth Disease Outbreaks in Ethiopia from 2010 to 2019

Foot-and-mouth disease (FMD) is an endemic disease in Ethiopia, although space&ndash;time cluster and monthly variation studies have never been assessed at national level. The current study aimed to identify the spatial and temporal distribution of FMD outbreaks in Ethiopia from national outbreak reports over a period of ten years from 1 January 2010 to 31 December 2019. To this end, a total of 376,762 cases and 1302 outbreaks from 704 districts were obtained from the Minister of Agriculture for analyses. In general, the dry periods, i.e., October to March, of the year were recorded as the peak outbreak periods, with the highest prevalence in March 2012. The monthly average and the outbreak trends over ten years show a decrease of outbreaks from 2010 to 2019. Decomposing the FMD outbreak data time series showed that once an outbreak erupted, it continued for up to five years. Only 12% of the reported outbreaks were assigned to a specific serotype. Within these outbreaks, the serotypes O, A, SAT-2, and SAT-1 were identified in decreasing order of prevalence, respectively. When a window of 50% for the maximum temporal/space cluster size was set, a total of seven FMD clusters were identified in space and time. The primary cluster with a radius of 380.95 km was identified in the southern part of Ethiopia, with a likelihood ratio of 7.67 (observed/expected cases). The third cluster, with a radius of 144.14 km, was identified in the northeastern part of the country, and had a likelihood ratio of 5.66. Clusters 1 and 3 occurred from January 2017 to December 2019. The second cluster that occurred had a radius of 294.82 km, a likelihood ratio of 6.20, and was located in the central and western parts of Ethiopia. The sixth cluster, with a radius of 36.04 km and a likelihood ratio of 20.60, was set in southern Tigray, bordering Afar. Clusters 2 and 6 occurred in the same period, from January 2014 to December 2016. The fourth cluster in northern Tigray had a calculated radius of 95.50 km and a likelihood ratio of 1.17. The seventh cluster occurred in the north-central Amhara region, with a radius of 97 km and a likelihood ratio of 1.16. Clusters 4 and 7 occurred between January 2010 and December 2013. The spatiotemporal and cluster analysis of the FMD outbreaks identified in the context of the current study are crucial in implementing control, prevention, and a prophylactic vaccination schedule. This study pointed out October to March as well as the main time of the year during which FMD outbreaks occur. The area that extends from the south to north, following the central highlands, is the main FMD outbreak area in Ethiopia

Additional file 1: of Isolation, molecular characterization and sero-prevalence study of foot-and-mouth disease virus circulating in central Ethiopia

Serological diagnostic tests procedures. Procedures and principles of the test during the study were discussed in detail. (DOCX 12 kb

Seroprevalence and risk factors of lumpy skin disease in Ethiopia

Lumpy skin disease (LSD) is an acute or inapparent viral disease of cattle which is endemic in many African and Middle East countries. LSD is one of the major transboundary livestock diseases in Ethiopia. A cross-sectional study using multistage cluster sampling was undertaken in central and north-western parts of Ethiopia with the objectives to estimate seroprevalence and to identify and quantify risk factors contributing to the occurrence of the disease. A total of 2386 cattle sera were sampled from 605 herds and 30 clusters (kebeles) located in 10 districts and tested for presence of LSD virus antibodies using virus neutralization test. All the serum samples were collected from cattle having no history of LSD vaccination. The overall animal level and herd level apparent seroprevalences were 25.4% (95% CI: 23.7–27.2) and 48.9% (95% CI: 44.9-52.9), respectively and varied significantly between districts. The true animal level and herd level prevalences were estimated as 26.5% (95% CI: 24.7–28.3) and 52.6% (95% CI: 48.3–56.9), respectively. At animal level, adult age (OR = 2.44 (95% CI: 1.67–3.55) compared to calf), contact with other animals (OR = 0.41 (95% CI: 0.23-0.74), compared to no contact) and presence of water bodies (OR = 1.61 (95% CI: 1.03–2.52), compared to no such bodies) were identified as the most important risk factors in relation to testing LSD positive. The putative risk factors altitude, breed, sex, and presence of animal trade route showed no significant association with LSD sero-status. Generally, cattle population with many adult animals and that live in wet areas are at highest risk, whereas cattle in frequent contact with other animals and animal species have lower risk, potentially due to a dilution effect of vectors.</p

Additional file 2: of Isolation, molecular characterization and sero-prevalence study of foot-and-mouth disease virus circulating in central Ethiopia

Antigen detection ELISA procedures. Antigen detection ELISA procedures and result interpretation during the study were discussed in detail. (DOCX 13 kb

Humoral Immune Response in Calves Vaccinated with Monovalent Vaccines or a Trivalent Combination Thereof and Matching of These Vaccines to the Selected Circulating Foot-and-Mouth Disease Viruses in Ethiopia

Foot-and-mouth disease (FMD) is an endemic, highly contagious, and devastating disease of livestock production in Ethiopia. Control of this disease relies mainly on prophylactic vaccination by willing farmers without a countrywide vaccination program. The objectives of this study were to quantify the humoral immune response and evaluation of the serological relationship of the vaccine strain used with representative field strain isolates. This was performed by primo vaccination of 6–9-month-old Holstein Friesian calves (35 treatment and 4 control calves) on day one and booster vaccination on day 28. Calves were vaccinated using the locally available National Veterinary Institute (NVI), Bishoftu, Ethiopia, inactivated aluminum hydroxide adjuvant monovalent (either O, A, SAT-2 alone) or trivalent (combination of A, O, SAT-2) vaccine (A/ETH/6/2000 (G-VII, O/ETH/38/2005(EA-3) and SAT-2/ETH/64/2009(XIII)). A 2 mL or 4 mL dose was used to vaccinate all calves except the animals that served as a control. In the case of the trivalent vaccine, a 4 mL dose was used to vaccinate calves. The serum was collected at 7, 14, 21, 28, and 56 days post-vaccination (d.p.v.). The humoral immune response was quantified by the solid-phase competitive enzyme-linked immunosorbent assay (SPC ELISA) and the virus-neutralization test (VNT). The serological relationship of heterologous and homologous viruses was also evaluated by adjuvant vaccine matching tests. The r1-value was determined using serum collected 21 d.p.v. An increase in immune response was observed from 7 d.p.v. to 28 d.p.v. in calves who received a 4 mL dose containing a 107.24 antigen load of 100 tissue culture infective dose (100TCID50) virus titer in the formulation. Upon receiving a booster dose on day 28, the humoral immune response was checked on the 56th day post-initial vaccination. Amounts of 54%, 72%, 79%, and 72% of inhibition for A, O, SAT-2, and trivalent vaccine in the three serotypes SPCE, respectively, was measured. Here, it was found that the immune response of calves increased from day 7 to 56, as evidenced by SPCE analysis. Likewise, an increase in antibody titer measured by a one-dimensional virus neutralization assay was also in line with SPCE analysis. This indicates that the vaccine is capable of inducing a neutralizing antibody that confers a protective immune response in 70%, 62%, and 100% heterologous field strains of A, O, and SAT-2 isolates, respectively, and has an average antigenic relationship of >0.3 with a standard deviation of +0.05 (N = 3) to the vaccine strains A/ETH/6/2000, O/ETH/38/2005 and SAT-2/ETH/64/2009, respectively, when using the one-dimensional virus neutralization test. The contribution and importance of this study is a confirmation of the vaccine and the field strain serological relationship for serotype SAT-2 strain and further research/change of vaccination strategy/ improvement in the currently used vaccine to cover a wide range of prevailing genotypes/lineages and induction of sound immune response after vaccination for serotype A and O strain. This study suggests that the trivalent vaccine produced by the National Veterinary Institute containing viral isolates from serotype O, A, and SAT-2 has a good serological relationship with the majority of circulating field strains in Ethiopia

Phenotypic, molecular detection and antibiogram analysis of Aeromonas Hydrophila from Oreochromis Niloticus (Nile Tilapia) and Ready-To- eat fish products in selected Rift Valley lakes of Ethiopia

Abstract Background Aeromonas hydrophila is a zoonotic bacterial pathogen that frequently causes disease and mass mortalities among cultured and feral fishes worldwide. In Ethiopia, A. hydrophila outbreak was reported in Sebeta fish ponds and in Lake Tana fishery. However, there is no to little information on the molecular, and phenotypical characteristics of A. hydrophila in Ethiopian fisheries. Therefore, a cross-sectional study was conducted from November 2020 to May 2021 in selected Ethiopian Rift valley lakes. Results A total of 140 samples were collected aseptically from fish (Muscle, Gill, Intestine, Spleen and Kidney) from fish landing sites, market and restaurants with purposive sampling methods. Aeromonas selective media (AMB), morphological and biochemical tests were used to isolate and identify A. hydrophila. Accordingly, the pathogen was isolated from 81 (60.45%) of samples. Among the isolates 92.59% expressed virulence trait through β hemolysis on blood agar media with 5% sheep blood. Moreover, 54 strains (66.67%) were further confirmed with Real-Time PCR (qPCR) using ahaI gene specific primers and optimized protocol. The highest (68.51%) were detected from live fish, (24.07%) were from market fish and the lowest (7.4%%) were from ready-to-eat products. Antibiogram analysis was conducted on ten representative isolates. Accordingly, A. hydrophila isolates were susceptible to ciprofloxacin (100%), chloramphenicol (100%) and ceftriaxone (100%). However, all ten isolates were resistant to Amoxicillin and Penicillin. Conclusions The study indicates A. hydrophila strains carrying virulence ahaI gene that were ß-hemolytic and resistant to antibiotics commonly used in human and veterinary medicine are circulating in the fishery. The detection of the pathogen in 140 of the sampled fish population is alarming for potential outbreaks and zoonosis. Therefore, further molecular epidemiology of the disease should be studied to establish potential inter host transmission and antibiotic resistance traits. Therefore, raising the public awareness on risk associated with consuming undercooked or raw fish meat is pertinent