Hygienic status of beef butcher shop facilities and antibiotic resistance profile of Salmonella enterica in Ethiopia.

The microbiological quality of meat is influenced by the conditions of hygiene prevailing during production and handling. Thus, this study aimed to assess the prevalence of Salmonella enterica and its antimicrobial resistance, load of hygiene indicator bacteria including E. coli (ECC), coliforms (CC), total coliform (TCC), Enterobacteriaceae (EB) and aerobic plate count (APC), and meat handler's food safety knowledge and hygiene practices in butcher shops in two cities, Addis Ababa and Hawassa in Ethiopia, during 2020 and 2021. A total of 360 samples of beef carcasses (n = 120), knives (n = 60), chopping boards (n = 60), weighing balance (n = 60), and personnel's hands (n = 60) were randomly collected for microbial analysis. Besides, 120 participants were selected to participate in a food safety knowledge and hygiene practices assessment. The S. enterica isolates were identified by agglutination test followed by qPCR targeting invA gene. Phenotypic antimicrobial resistance profiles of S. enterica were determined using disk diffusion assays as described in CLSI. The ECC, CC, TCC, EB, and APC populations were quantified by plating onto petrifilm plates. A structured questionnaire was used to determine food safety knowledge and hygiene practices of participants. Overall prevalence of S. enterica was 16.7% (95% CI, 8.3-26.7) and location seems to have no effect (p = 0.806). Only 20% of the S. enterica were resistant to ampicillin and tetracycline. However, the majority (80%) of S. enterica isolates were susceptible to the panel of 11 antimicrobials tested. The overall mean ± SD (log CFU/cm2) of ECC, CC, TCC, EB, and APC were 4.31 ± 1.15; 4.61 ± 1.33; 4.77 ± 1.32; 4.59 ± 1.38 and 5.87 ± 1.52, respectively. No significant difference (p = 0.123) in E. coli contamination was observed between samples of beef carcasses and chopping boards. The EB contamination showed no significant difference (p > 0.05) among sample sources. The APC contamination levels on beef carcass were significantly higher (p > 0.05) than other sample sources. A total of 56% (95% CI: 46.7 - 65.0) of the participants had poor knowledge and 65% (95% CI: 56.7 - 73.3) had poor hygiene practices towards food safety. This study highlighted the poor hygiene status of butcher facilities with a potential risk of beef safety. Thus, appropriate food safety control strategies and inspection is needed at retail establishments.

Antimicrobial Resistance of Escherichia coli Isolates from Livestock and the Environment in Extensive Smallholder Livestock Production Systems in Ethiopia.

The objective of this study was to characterize the distribution of antimicrobial resistance (AMR) of Escherichia coli (E. coli) isolated from livestock feces and soil in smallholder livestock systems. A cross-sectional study was carried out sampling 77 randomly selected households in four districts representing two agroecologies and production systems. E. coli was isolated and the susceptibility to 15 antimicrobials was assessed. Of 462 E. coli isolates tested, resistance to at least one antimicrobial was detected in 52% (43.7-60.8) of isolates from cattle fecal samples, 34% (95% CI, 26.2-41.8) from sheep samples, 58% (95% CI, 47.9-68.2) from goat samples and 53% (95% CI, 43.2-62.4) from soil samples. AMR patterns for E. coli from livestock and soil showed some similarities, with the highest prevalence of resistance detected against streptomycin (33%), followed by amoxycillin/clavulanate (23%) and tetracycline (8%). The odds of detecting E. coli resistance to ≥2 antimicrobials in livestock fecal samples were nearly three times (Odd Ratio-OR: 2.9; 95% CI, 1.72-5.17; p = 0.000) higher in lowland pastoral than in highland mixed crop-livestock production systems. These findings provide insights into the status of resistance in livestock and soil, and associated risk factors in low-resource settings in Ethiopia.

Animal Brucellosis: Seropositivity rates, Isolation and Molecular Detection in Southern and Central Ethiopia.

Introduction: Brucellosis is a neglected bacterial zoonosis with serious veterinary and public health importance throughout the world. A cross-sectional study on animal brucellosis was conducted aiming to estimate seroprevalence and molecular detection. Methods: Blood samples were collected from a total of 4274 individual animals (cattle, small ruminants and camel) from 241 herds/flocks for serology and PCR. Serum samples were tested using multispecies I-ELISA. Blood clots from seropositive animals were also tested for brucellosis via PCR. Additionally, 13 vaginal swab samples were collected from animals (2 from bovine and 11 from small ruminants) with recent abortion history for bacterial isolation and molecular detection. Results: The overall individual animal and herd level seroprevalence was 3.95% (169/4274) and 18.26% (44/241) respectively. The animal level seroprevalence at species level was 1.58% (47/2982), 8.89% (97/1091) and 12.44% (25/201) in bovine, small ruminants (sheep and goat) and camel, respectively. Herd level seroprevalence were 5.43% (10/184), 52.08% (25/48) and 100% (9/9) in bovine, small ruminant and camel, respectively. The animal level seroprevalence of bovine from intensive and extensive systems was 1.10% (31/2808) and 2.87% (5/174) respectively. Blood clots tested for brucellosis via PCR were negative by RT-PCR. Brucella species was isolated from 6/13 (46.15%) vaginal swab samples cultured on Brucella selective agar, and shown to be B. melitensis using Real-Time PCR. Conclusion: Overall, seropositivity for camels was higher than what has been reported previously. Also, there was a notable difference in this study in cattle seroprevalence when comparing extensive with intensive systems, with the extensive system having much greater seropositivity.

Utility of the Intradermal Skin Test in a Test-and-Cull Approach to Control Bovine Tuberculosis: A Pilot Study in Ethiopia.

Bovine tuberculosis (bTB) is one of the top three, high-priority, livestock diseases in Ethiopia and hence, the need for evaluation of potential control strategies is critical. Here, we applied the test-and-segregate followed by cull strategy for the control of bTB in the intensive Alage dairy farm in Ethiopia. All cattle reared on this farm were repeatedly skin tested using the Comparative Cervical Tuberculin (CCT) test for a total of five times between 2015 and 2021. During the first (October 2015) and second (March 2017) rounds of testing, all reactor animals (>4 mm) were culled, while those that were deemed as inconclusive (1-4 mm) were segregated and retested. At retest, animals with CCT >2 mm were removed from the herd. In the third (December 2017) and fourth (June 2018) rounds of tuberculin testing, a more stringent approach was taken wherein all reactors per the severe mode of CCT test interpretation (>2 mm) were culled. A final herd status check was performed in May 2021. In summary, the number of CCT positives (>4 mm) in the farm dropped from 23.1% (31/134) in October 2015 to 0% in December 2017 and remained 0% until May 2021. In contrast, the number of Single Cervical Tuberculin (SCT) test positives (≥4 mm) increased from 1.8 to 9.5% (from 2017 to 2021), indicating that CCT test might not be sufficient to effectively clear the herd of bTB. However, a more stringent approach would result in a drastic increase in the number of false positives. The total cost of the bTB control effort in this farm holding 134-200 cattle at any given time was conservatively estimated to be ~US$48,000. This, together with the need for culling an unacceptably high number of animals based on skin test status, makes the test-and-cull strategy impractical for nationwide implementation in Ethiopia and other low- and middle-income countries (LMICs) where the infection is endemic. Hence, there is an increased emphasis on the need to explore alternate, affordable measures such as vaccination alongside accurate diagnostics to help control bTB in endemic settings.

Spoligotype analysis of Mycobacterium bovis isolates from cattle and assessment of zoonotic TB transmission among individuals working in bovine TB-infected dairy farms in Ethiopia.

Bovine tuberculosis (bTB) is a disease with impact on dairy productivity, as well as having the potential for zoonotic transmission. Understanding the genetic diversity of the disease agent Mycobacterium bovis is important for identifying its routes of transmission. Here we investigated the level of genetic diversity of M. bovis isolates and assessed the zoonotic potential in risk groups of people working in bTB-infected dairy farms in central Ethiopia. M. bovis was isolated and spoligotyped from tissue lesions collected from slaughtered cattle as well as from raw milk collected from bTB positive cows in dairy farms from six urban areas of central Ethiopia. From consented dairy farm workers, knowledge and practices related to zoonotic TB transmission, together with demographic and clinical information, was collected through interviews. Sputum or Fine Needle Aspirate (FNA) samples were collected from suspected TB cases. Spoligotyping of 55 M. bovis isolates that originated either from cattle tissues with tuberculous lesion or from raw milk revealed seven spoligotype patterns where SB1176 was the most prevalent type (47.3%). Most isolates (89.1%) were of the M. bovis African 2 clonal complex. All sputum and FNA samples from 41 dairy farm workers with symptoms of TB were culture negative for any mycobacteria. Among the 41 TB suspected farm workers, 61% did not know about bTB in cattle and its zoonotic potential, and over two-third of these workers practiced raw milk consumption. Our spoligotype analysis suggests a wider transmission of a single spoligotype in the study area. The results reported here may be useful in guiding future work to identify the source and direction of bTB transmission and hence design of a control strategy. Isolation of M. bovis from milk, knowledge gap on zoonotic TB and practice of consumption of raw milk in the study population showed potential risk for zoonotic transmission.

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.

Population structure and transmission of Mycobacterium bovis in Ethiopia.

Bovine tuberculosis (bTB) is endemic in cattle in Ethiopia, a country that hosts the largest national cattle herd in Africa. The intensive dairy sector, most of which is peri-urban, has the highest prevalence of disease. Previous studies in Ethiopia have demonstrated that the main cause is Mycobacterium bovis, which has been investigated using conventional molecular tools including deletion typing, spoligotyping and Mycobacterial interspersed repetitive unit-variable number tandem repeat (MIRU-VNTR). Here we use whole-genome sequencing to examine the population structure of M. bovis in Ethiopia. A total of 134 M. bovis isolates were sequenced including 128 genomes from 85 mainly dairy cattle and six genomes isolated from humans, originating from 12 study sites across Ethiopia. These genomes provided a good representation of the previously described population structure of M. bovis, based on spoligotyping and demonstrated that the population is dominated by the clonal complexes African 2 (Af2) and European 3 (Eu3). A range of within-host diversity was observed amongst the isolates and evidence was found for both short- and long-distance transmission. Detailed analysis of available genomes from the Eu3 clonal complex combined with previously published genomes revealed two distinct introductions of this clonal complex into Ethiopia between 1950 and 1987, likely from Europe. This work is important to help better understand bTB transmission in cattle in Ethiopia and can potentially inform national strategies for bTB control in Ethiopia and beyond.

Genotype Diversity of Mycobacterium bovis and Pathology of Bovine Tuberculosis in Selected Emerging Dairy Regions of Ethiopia.

Bovine tuberculosis (bTB) is endemic in Ethiopia with higher prevalence in cattle, particularly in the central parts. Spread of Mycobacterium bovis (M. bovis) to wider regions is inevitable in uncontrolled conditions. This study was conducted to explore the pathology, characterize M. bovis strains, and describe genotypic diversity to demonstrate possible epidemiological links in emerging dairy areas of Ethiopia, namely, Mekelle and Gondar. Twenty-seven bTB positive cattle identified by the Single Intradermal Comparative Cervical Tuberculin (SICCT) test were subjected to post-mortem inspection to determine lesion distribution and pathological score. Samples from tissues with visible tuberculous or suspected non-visible lesions were processed and cultured following a standard protocol. Isolates identified as M. bovis by Region of Difference (RD)-based Polymerase Chain Reaction (PCR) were also spoligotyped to determine their spoligotype patterns. Post-mortem inspection of visceral organs indicated bTB suggestive lesions in 41% of the animals, with 25% being in the lungs. Lymph nodes from 77% of the animals had lesions. Fifty-five isolates identified from 24 of the slaughtered animals were confirmed as M. bovis. No other mycobacterial species were isolated. Spoligotyping classified strains from 21 of these animals into seven spoligotype patterns: SB0133, SB0134, SB1176, SB2233, SB2290, SB2467, and SB2520. More than one spoligotype were identified from five of these animals, and none of the last four spoligotypes had been reported in Ethiopia before. SB0134 was the most predominant type (47%) followed by SB0133 (25.5%). SB0133, SB2290, SB2467, and SB1176 are spoligotypes lacking spacers 3-7, characteristics of M. bovis strains of the African 2 (Af2) clonal complex, while SB0134, SB2233, and SB2520 do not belong to any of the established clonal complexes and likely to have a different evolutionary history. Despite a small sample size, the present study showed strain diversity with multiple genotypes identified in a single herd and even within a single animal, and the genotypes showed no sign of geographical localization, which could be a consequence of significant movement of bTB diseased cattle around the country, spreading the disease. Therefore, any future control programme of bTB in Ethiopia needs to address the risks of cattle movement.

Survival Analysis of Clinical Cases of Caseous Lymphadenitis of Goats in North Shoa, Ethiopia.

Caseous lymphadenitis (CLA) is a bacterial disease of small ruminants causing abscesses in lymph nodes of the body and internal organs. A longitudinal study from 2011 to 2019 was undertaken at Ataye site of Debre Birhan Research Center to estimate the prevalence and incidence, identify associated risk factors, and characterize the causative bacteria. 1025 goats were followed up for the CLA study. Survival analysis was done using SAS 9.4 software program. Biochemical tests and Biolog microbial identification system were used to characterize the bacteria. In the recurrent survival data analysis, there were 1,138 clinical observations and 214 CLA cases (18.8% prevalence) based on clinical diagnosis. The incidence rate was 0.14 cases per animal year. From a total of 214 cases, 130 have occurred once and 84 cases are recurrent cases following first cases. The cumulative failure rates were 68% for newborn and 64% for yearling age groups. The parotid lymph node was the most frequently affected site. Proportional hazard analysis results showed that sex, age, and breed were associated (p < 0.05) with CLA incidence. Females, newborn age group, Boer, and Boer × Central Highland Goat (CHG) were at higher risks compared to males, yearling age group, and CHG goats, respectively. The present study indicated that a high incidence rate of CLA in the goat farm is difficult to control and prevent because of its poor response to clinical treatment. Thus, control and prevention should focus on the spread of the disease such as isolation of clinically sick animals, culling, and vaccination of flocks.

Antibiograms and risk factors of Salmonella isolates from laying hens and eggs in Jimma Town, South Western Ethiopia.

OBJECTIVES: Salmonella is the most important causes of foodborne illness especially from poultry and poultry products. So the aim of this study was to carryout phenotypic characterization, antimicrobials susceptibility pattern and risk factors of Salmonella isolates from farms and markets eggs, cloacae swabs of chickens and stool of egg collectors. A cross-sectional study was conducted from January 2018 to September 2018. Samples were, processed; Salmonella was isolated, phenotypically identified by OmniLog and antimicrobials susceptibility were carried out. RESULT: Over all; 11 (2.65%) of Salmonella enterica were phenotypically characterized out of 415 samples from farms egg content (n = 83), farms eggshell (n = 83), cloacae (n = 83), market eggshell (n = 83) and market egg contents (n = 83) with 2.4%, 0%, 2.4%, 4.8% and 3.6% prevalence, respectively. Out of isolates, 8 (72.72%) displayed multidrug resistance. All isolates showed susceptibility to Gentamicin, Kanamycin and Streptomycin. Lack of separating cracked eggs, washing hand, eggs stay longer unsold, and mixing excreta with feed were associated risk factors for Salmonella presence (P-value < 0.05). The presence of drug resistant Salmonella enterica within egg/and chicken can pose serious health problem. Good hygienic practices are important to reduce risk factors of Salmonella contamination.

Prevalence of camel trypanosomosis (surra) and associated risk factors in Borena zone, southern Ethiopia.

A study was made to determine the prevalence of camel trypanosomosis (surra) and its associated risk factors in Borena zone, southern Ethiopia during 2013-2014. A total of 2400 blood samples were collected and examined by the buffy coat and thin blood film laboratory methods, and data were analyzed using the SPSS statistical software. The overall prevalence of camel trypanosomosis in the area was found to be 2.33 %. Prevalence was significantly different among the surveyed districts (P = 0.000), the pastoral associations (F = 6.408, P = 0.000), altitudinal divisions (P = 0.000), age groups (P = 0.034), and between animals possessing packed cell volume (PCV) values greater than 25 % and less than 25 % (P = 0.000); whereas, prevalence of the disease was not statistically significantly different between the sexes (P = 0.311) and among the body condition score groups (P = 0.739). The PCV of trypanosome positive and trypanosome negative camels differ significantly (P = 0.001), and prevalence of trypanosomosis was seen to be negatively correlated with packed cell volume (r = -0.069, P = 0.000) revealing the effect of camel trypanosomosis on anemia state of parasitized animals. In conclusion, camel trypanosomosis is a serious and economically important disease hampering camel production and productivity in southern Ethiopia. Further studies involving more sensitive molecular techniques to reveal the precise magnitude of the disease and to identify the vector species of the parasite are recommended.

Spatial distribution of Glossina sp. and Trypanosoma sp. in south-western Ethiopia.

BACKGROUND: Accurate information on the distribution of the tsetse fly is of paramount importance to better control animal trypanosomosis. Entomological and parasitological surveys were conducted in the tsetse belt of south-western Ethiopia to describe the prevalence of trypanosomosis (PoT), the abundance of tsetse flies (AT) and to evaluate the association with potential risk factors. METHODS: The study was conducted between 2009 and 2012. The parasitological survey data were analysed by a random effects logistic regression model, whereas the entomological survey data were analysed by a Poisson regression model. The percentage of animals with trypanosomosis was regressed on the tsetse fly count using a random effects logistic regression model. RESULTS: The following six risk factors were evaluated for PoT (i) altitude: significant and inverse correlation with trypanosomosis, (ii) annual variation of PoT: no significant difference between years, (iii) regional state: compared to Benishangul-Gumuz (18.0%), the three remaining regional states showed significantly lower PoT, (iv) river system: the PoT differed significantly between the river systems, (iv) sex: male animals (11.0%) were more affected than females (9.0%), and finally (vi) age at sampling: no difference between the considered classes. Observed trypanosome species were T. congolense (76.0%), T. vivax (18.1%), T. b. brucei (3.6%), and mixed T. congolense/vivax (2.4%). The first four risk factors listed above were also evaluated for AT, and all have a significant effect on AT. In the multivariable model only altitude was retained with AT decreasing with increasing altitude. Four different Glossina species were identified i.e. G. tachinoides (52.0%), G. pallidipes (26.0%), G.morsitans submorsitans (15.0%) and G. fuscipes fuscipes (7.0 %). Significant differences in catches/trap/day between districts were observed for each species. No association could be found between the tsetse fly counts and trypanosomosis prevalence. CONCLUSIONS: Trypanosomosis remains a constraint to livestock production in south-western Ethiopia. Four Glossina and three Trypanosoma species were observed. Altitude had a significant impact on AT and PoT. PoT is not associated with AT, which could be explained by the importance of mechanical transmission. This needs to be investigated further as it might jeopardize control strategies that target the tsetse fly population.