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Livestock, farms, abattoirs, and food supply systems can become the source of foodborne pathogens, including S. aureus , in the absence of monitoring, general hygienic practices, and control. Studies are scarce on reservoirs (hiding places) and routes of entry of S. aureus into the food supply chain in Ethiopia. To fill these gaps, we evaluated the role of cows (milk), meat, equipment, and food handlers on the abundance and AMR of S. aureus in five geographical areas in central Oromia, Ethiopia. We isolated S. aureus from 10 different ecologies per area in 5 areas and tested their sensitivity to 14 antimicrobials of 9 different classes. We ranked the 5 areas and 10 ecologies by computing their multiple AMR index (MARI) at a cut-off value of 0.2 to determine 'high-risk' ecologies for AMR. We recorded as MDR if an isolate had resistance to ≥3 antimicrobial classes. We used a circos diagram to analyze if isolates with identical AMR patterns were shared between different ecologies. S. aureus is prevalent in central Oromia (16.1-18.3%), higher in dairy farms than in abattoirs, and varied among 10 ecologies ( p < 0.001) but not among 5 areas ( p > 0.05). Of the 92 isolates, 94.6% were penicillin-resistant. Their AMR prevalence was above 40% for 9 of 14 antimicrobials. All isolates (100%) had AMR in at least one antimicrobial class (range = 1-9; median = 5), indicating MDR was prevalent. The prevalence of MDR S. aureus varied ( p < 0.05) among areas and 10 ecologies; the highest was in slaughter lines. All isolates had a MARI of >0.2, indicating drug overuse, and S. aureus's AMR burden is high in central Oromia. Dairy farms had higher MARI values (0.44) than abattoirs (0.39). Of 10 ecologies, the highest and lowest MARI values were in the beef supply chain, i.e., slaughter line (0.67) and butcher's hand (0.25). Of the 68 different AMR patterns by 92 isolates against 14 antimicrobials, 53 patterns (77.9%) were unique to individual isolates, indicating they were phenotypically dissimilar. MDR S. aureus was widespread in central Oromia in dairy and meat supply chains, contaminating milk, meat, equipment, and workers in farm and abattoir settings. In the absence of strict regulations and interventions, MDR S. aureus can be disseminated from these epicenters to the public.

Background: In veterinary medicine, three Staphylococcus species are of particular importance as primary causes of specific diseases; S. aureus (mastitis in ruminants, equine botryomycosis, and bumble foot in poultry), S. hycus (porcine exudative epidermitis), and S. intermedius (canine pyoderma). The disease conditions caused by Staphylococcus in poultry vary with site, route, and predisposing factors include wounds as a result of fighting/cannibalism, immunosuppression based on virus infection or parasite infestation, and bad husbandry conditions (overcrowding). The objectives of this study were to isolate and identify Staphylococcus spp from chicken and chicken litter and personnel at chicken farm and to determine the antimicrobial susceptibility profile of the isolates., Methods: A cross-sectional study was conducted on apparently healthy chickens, farm personnel, and chicken litter at poultry farms in Addis Ababa, Ethiopia. A total of 222 samples consisting of 101 cloacal swabs, 90 tracheal swabs, 17 pooled litter swabs, 7 nasal swabs, and 7 pooled hand and boot swabs were collected from six farms and examined for the presence of Staphylococcus species. Antimicrobial resistance against 10 antimicrobial agents was also conducted following recommended standard procedures., Results: Overall proportion of Staphylococcus was 64/222 (28.83%). Of the isolates, 40/64 (62.5%), 11/64 (17.2%), 3/64 (4.7%), and 10/64 (15.6%), were S. aureus , S. hycus , S. intermedius , and coagulase-negative staphylococci (CNS), respectively. Only one isolate of S. aureus was susceptible to all antimicrobials tested. Of the 10 antibiotics tested, the isolates demonstrated highest resistance against Penicillin G (96.9%) followed by Tetracycline (78.1%), and Amoxicillin and Erythromycin at the same level (65.6%). Conversely, the isolates were highly susceptible to Ciprofloxacin (95.3%) followed by Sulphamethoxazole-trimethoprim (85.9%). Out of 64 isolates, 61/64 (95.3%) were resistant to three or more antimicrobials tested. Of the isolates, 38/40 (95%) S. aureus, 10/11 (90.9%) S. hycus , 3/3 (100%) S. intermedius , and 10/10 (100%) CNS showed multidrug resistance., Conclusion: This study showed a considerable proportion of Staphylococcus spp in chicken litter and farm workers with a potential source of resistant Staphylococcus species, and more importantly multidrug resistance strains. Further studies on molecular characterization of the isolates will be essential to identify the resistant genes and establish epidemiological links in the transmission dynamics of resistant Staphylococcus species between poultry and humans., Competing Interests: The authors declare that they have no conflicts of interests., (Copyright © 2022 Fufa Abunna et al.)

The Ethiopian government has initiatives for expanding the commercial and smallholder market-oriented urban and peri-urban dairy production systems to meet the demands for dairy products. However, there have been only limited on-farm studies on the health performance of commercial dairy breeds. The aim of this longitudinal study was to quantify the incidence and identify predictors of calf morbidity and mortality from birth to 6 months of age in urban and peri-urban dairy farms of Northwest Ethiopia. A total of 439 calves aged below 6 months from 174 dairy farms were included in the study. We collected data on 35 potential risk factors to determine their effect on calf morbidity and mortality in the area. Kaplan-Meier survival analysis was used to summarize survival probability. The Cox proportional hazard regression model with shared frailty to account for unmeasured herd-specific heterogeneity was also used to identify and quantify factors associated with time to morbidity and mortality. Among 439 calves enrolled for 6 months of follow-up period, a total of 141 morbidities and 54 mortality events were recorded. This gives an overall morbidity and mortality incidence rates of 64 per 100-calf 6-months at risk (risk rate of 47.3%) and 19 per 100-calf 6-months at risk (risk rate of 17.9%), respectively. Diarrhea was the most frequent calf health problem with a risk rate of 25.2%. It was the cause of death for 33.3% of all the 54 calf deaths. Next to diarrhea, pneumonia ranked second with risk rate of 8.6% and was responsible for death of 12.9% of all the 54 calf deaths. Among 35 potential risk factors, calf age, vigor status at birth, calf breed, colostrum ingestion, and herd size were significant ( p < 0.05) predictors of calf morbidity and mortality. The Cox-shared frailty model revealed that the herd frailty component had no significant effect on hazard estimates of the covariates of all-cause morbidity and mortality. This implies that the dairy herds participated in the study were homogeneous in the distribution of unmeasured random effects. In conclusion, the magnitude of calf morbidity and mortality was higher and above economically tolerable level in this study. This could impede the success of Ethiopia's dairy development initiative in general, and the livelihood of smallholder dairy producers in particular. Therefore, educating farmers aimed at mitigating the identified risk factors can reduce calf morbidity and mortality in the study areas., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Alemu, Jemberu, Mekuriaw and Abdi.)

Within Ethiopia, there is a lack of information on the genetic relatedness of Salmonella from cattle, beef, and diarrheic patients and its potential transmission from cattle to humans through consumption of contaminated beef. The objective of this study was to assess the prevalence and determine the serotypes, genetic relatedness, and antimicrobial resistance of Salmonella in cattle in two local slaughterhouses, in beef at retail shops, and in diarrheic patients in the only hospital in Bishoftu, Ethiopia. Salmonella was detected in 2.5% (6/240) of cattle samples, in 8.7% (11/127) of beef samples, and in 2.3% (5/216) of the diarrheic patients. Four Salmonella serotypes: Salmonella Typhimurium, Salmonella Eastbourne, Salmonella Saintpaul, and Salmonella Cotham were identified. Salmonella Typhimurium and Salmonella Eastbourne were isolated from cattle and beef, whereas Salmonella Saintpaul and Salmonella Cotham were isolated only from diarrheic patients. Except for serotype Salmonella Saintpaul, all isolates were grouped into five pulsotypes, of which two pulsotypes contained isolates from cattle and beef. Isolates from humans represented unique pulsotypes. Among the 22 Salmonella isolates tested, 95.5% were resistant to at least 1 of the 14 antimicrobials tested. Three Salmonella isolates originating from cattle were multidrug resistant. One human isolate was susceptible to all antimicrobials tested. More specifically, resistance to ampicillin, sulfamethoxazole, tetracycline, tigecycline, and trimethoprim were observed. The most frequently observed resistance was to sulfamethoxazole (90.9%, 20/22) followed by trimethoprim (22.7%, 5/22). The study revealed considerable Salmonella contamination of beef at retail shops, antimicrobial resistance to commonly used antimicrobials, and shared genetically similar Salmonella serotypes between cattle and beef; the link with humans could not be established. Still, the findings of Salmonella in cattle and beef, the propensity of transfer of Salmonella from cattle to beef coupled with the common consumption of raw/undercooked beef are likely to pose public health risk in Ethiopia.

Understanding the potential drivers of microbial meat contamination along the entire meat supply chain is needed to identify targets for interventions to reduce the number of meatborne bacterial outbreaks. We assessed the hygienic practices in cattle slaughterhouses (28 employees) and retail shops (127 employees) through face-to-face interviews and direct personal observations. At the slaughterhouses, stunning, de-hiding and evisceration in vertical position, carcass washing and separate storage of offal were the identified good practices. Lack of hot water baths, absence of a chilling room, infrequent hand washing, insufficiently trained staff and irregular medical check-up were practices that lead to unhygienic handling of carcasses. At the retail shops, cleaning equipment using soap and hot water (81%), storing unsold meat in refrigerators (92%), concrete floors and white painted walls and ceilings were good practices. Adjacently displaying offal and meat (39%), lack of a cold chain, wrapping meat with plastic bags and newspapers, using a plastic or wooden cutting board (57%), infrequent washing of equipment and floors, and inadequately trained employees were practices that could result in unhygienic handling of beef. Our study identified unhygienic practices both at the slaughterhouses and retail shops that can predispose the public to meatborne infections, which could be improved through training and implementation of quality control systems.

Mastitis is the most prevalent and economically important disease caused by different etiological agents, which leads to increased somatic cell count (SCC) and low milk quality. Treating mastitis cases with antimicrobials is essential to reduce SCC and improve milk quality. Non-prudent use of antimicrobials in dairy farms increased the development of antimicrobial resistant bacteria. This study's objectives were (1) to isolate and identify etiological agents of mastitis and (2) to determine antimicrobial resistance profiles of bacterial isolates. A total of 174 quarter milk samples from 151 cows with high SCC and clinical mastitis from 34 dairy farms in Tennessee, Kentucky, and Mississippi were collected. Bacterial causative agents were determined by bacteriological and biochemical tests. The antimicrobial resistance of bacterial isolates against 10 commonly used antimicrobials was tested. A total of 193 bacteria consisting of six bacterial species, which include Staphylococcus aureus , Streptococcus uberis , Streptococcus dysgalactiae , Escherichia coli , Klebsiella oxytoca and Klebsiella pneumoniae were isolated. Staphylococcus aureus was the predominant isolate followed by Strep. spp., E. coli , and Klebsiella spp. Results of this study showed that Gram-negatives ( E. coli and Klebsiella spp.) were more resistant than Gram-positives ( Staph. aureus and Streptococcus spp.). Continuous antimicrobial resistance testing and identification of reservoirs of resistance traits in dairy farms are essential to implement proper mitigation measures.

Escherichia coli O157 is a Shiga toxin-producing E. coli causing disease in humans. Cattle are the primary reservoir of the pathogen. Information regarding the contribution of cattle to diarrheal illnesses in humans through consumption of contaminated beef is scarce in Ethiopia. We collected samples from 240 cattle, 127 beef, and 216 diarrheic patients in Bishoftu town in Ethiopia to assess the occurrence and determine the virulence genes, genetic relatedness, and antimicrobial resistance of E. coli O157. E. coli O157 was detected in 7.1% of the rectal content samples from cattle in slaughterhouses, in 6.3% ( n  = 127) of the beef samples, and in 2.8% of the diarrheic patients' stool samples. All isolates were positive for eae gene, 24 (77%) of them were positive for stx 2 gene (21 stx 2c and 3 stx 2a), whereas stx 1 gene was not detected. Molecular typing grouped the isolates into eight pulsed-field gel electrophoresis pulsotypes with three pulsotypes containing isolates from all three sources, one pulsotype containing one isolate from human origin and one isolate from beef. The remaining four pulsotypes contained isolates unique either to beef or to humans. With the exception of 1 multidrug-resistant isolate from beef, which was resistant to 8 antimicrobial drugs, the remaining 30 isolates were susceptible to the 14 antimicrobials tested. In conclusion, the finding of genetically similar isolates in cattle, beef, and humans may indicate a potential transmission of E. coli O157 from cattle to humans through beef. However, more robust studies are required to confirm this epidemiological link.

Dogs are reservoirs of many zoonotic diseases. In Ethiopia, the majority of owned dogs are semi-stray, freely roaming in the community. Studies reporting dog borne zoonotic diseases are scarce in Ethiopia. This study was conducted to assess Dipylidium caninum infection in dogs and in children with gastrointestinal complaints in Bishoftu Town, Oromia. We collected 384 fecal samples from dogs presented to veterinary teaching hospital and 259 stool samples from children presented to Bishoftu Hospital for clinical examination. Samples were first macroscopically examined for the presence of proglotids, followed by microscopic examination for the presence of eggs with the direct smear following flotation technique. The prevalence of D. caninum was 21% (95% CI: 16.6-24.9) in dogs. Although not statistically significant ( p > 0.05), higher prevalence was detected in adult (11.9%), local breed (17.7%), and male (12.6%) dogs compared to young (8.59%), exotic breed (2.86%), and females (7.81%), respectively. Dipylidium caninum was detected in a stool sample obtained from a three year-old child (0.4%, 1/259). This study showed that the prevalence of D. caninum in the dogs is high while it is rare in children. Although the prevalence in children is negligible in this study, the high proportion of infected dogs can pose a significant risk of infection in the general human population. Public health risk can be reduced by eliminating the semi-roaming of owned dogs and proper management of dogs with regular deworming and prevention of environmental contamination with dog feces. Similarly, raising public awareness about dog borne zoonoses and avoiding contact with dog feces are important.

Background: Bovine cysticercosis is a worldwide zoonotic disease that affects cattle caused by the larval stage of the tapeworm Taenia saginata, the adult parasite that causes taeniasis in humans. Although bovine cysticercosis, and the associated human taeniasis, is controlled in developed countries, it is one of the neglected tropical diseases. Like other parts of Ethiopia, raw or undercooked beef consumption is common in the rural community of Yem district with no meat inspection services., Methods: We conducted an abattoir survey to estimate the prevalence of bovine cysticercosis in cattle and a questionnaire survey to determine the level of historical human taeniasis infection in Yem district of Ethiopia., Results: Bovine cysticercosis was detected in 3.1% of 485 cattle examined. Although animal level prevalence was low, a subset of positive animals had a higher rate of cysticercosis infection widely distributed in various parts of the body. Over two-fifths (40%) of the cysts were found in the tongue making it an important organ to look for during meat inspection. Over half of the cysts were viable indicating their potential to infect people. History of taeniasis was reported by 71.5% of 151 residents interviewed during the study. Raw meat consumption was very common (89.4%) among the residents; people who consumed raw meat were 25 times more likely to report taeniasis than those who did not. In addition, traditional treatments using herbs and chemicals of unknown efficacy and risk were very common., Conclusions: Bovine cysticercosis and human taeniasis are important in this rural community. Providing meat inspection services and public health education on intervention measures such as proper cooking of meat, access to latrines and clean water are effective strategies that can break the lifecycle of the parasite and ensure beef safety and public health., (© 2020 Blackwell Verlag GmbH.)

Staphylococcus aureus is one of the major bacterial mastitis pathogens with significant effects on animal and human health. Some studies showed that S. aureus strains that infect different host species are genetically distinct, although most strains can infect a wide range of host species. However, there are no clearly defined clonal patterns of S. aureus strains that are known to infect a specific host. The objectives of this study were to evaluate the clonal diversity and virulence characteristics of S. aureus isolates from cases of bovine mastitis. Bacteriological tests were conducted on milk samples from cases of bovine mastitis from 11 dairy farms including some milk samples from unknown farms in Eastern Tennessee. Overall, a total of 111 S. aureus were isolated and identified, and further evaluated for their genetic diversity by pulsed-field gel electrophoresis (PFGE) and virulence characteristics by PCR. Genotypic virulence factors including staphylococcal enterotoxins, and toxic shock syndrome toxin 1 (tsst-1) were tested by PCR. In addition, the association among several known virulence factors of these isolates based on our current and previous studies in our lab were evaluated. Previously generated data that were included in the analysis of association among virulence factors were the presence of biofilm production associated genes in the ica operon such as icaA, icaD and icaAB, and phenotypic virulence characteristics such as hemolysis on blood agar, slime production and resistance or susceptibility to ten commonly used antimicrobials in dairy farms. The PFGE results showed the presence of 16 PFGE types (designated A - P) throughout farms, of which three pulsotypes, I, M and O were the most frequently isolated PFGE types from most farms. The PFGE type M was the most prevalent of all 16 PFGE types, with 64 isolates being present among nine farms. The PCR results of enterotoxin genes showed that out of the total 111 tested 84 (75.7%) were negative whereas 13 (11.7%), 2 (1.8%), 3 (2.7%), 1 (0.9%) and 8 (7.2%) were positive for seb, seb and sec, sec, see, and tsst-1, respectively. All 111 isolates were negative for sea and sej. Results of the evaluation of I, M and O strains adhesion to and invasion into mammary epithelial cells showed that the total count of each strain of bacteria adhered to and invaded into mammary epithelial cell line (MAC-T cells) was not significantly different (P > 0.05). This may be an indication that there is no significant difference in their ability to establish early host-pathogen interaction and colonization of the host. There were no statistically significant associations among PFGE types and other known virulence factors of these strains. However, PFGE types O and M tend to cluster with β-hemolysin, absence of enterotoxins and susceptibility to antimicrobials. In conclusion, there was not any association between pulsotype and genotypic and phenotypic virulence factors. S. aureus isolates from cases of bovine mastitis had diverse genotypes that possessed variable virulence factors., Competing Interests: Declaration of competing interest No competing financial interests exist for all authors., (Published by Elsevier Ltd.)

Background: Safe food is central to social wellbeing. Coagulase-negative staphylococci (CNS) are a threat to food safety because they may harbor multiple enterotoxins and antimicrobial resistance (AMR) genes. CNS bacteria are an emerging nosocomial pathogen in public health. CNS also cause bovine mastitis with a significant economic loss in the dairy industry and may introduce toxins to the food supply chain resulting in foodborne illnesses. However, information on CNS and their AMR status are scarce in food animal production and processing lines in Ethiopia., Methodology: This cross-sectional study evaluated the prevalence and AMR patterns of CNS in dairy farms and abattoirs using samples (n = 1001) from udder milk, beef carcass, personnel, and different abattoir and dairy equipment across five locations of central Oromia. The CNS isolates were identified via standard microbiological protocols and evaluated using disc diffusion test against 14 antimicrobials belonging to nine different broad classes. Uni-and-multivariable logistic regressions were used to analyze the association between potential risk factors (location, sample source, and sample type) and positivity to CNS., Results: The overall prevalence of CNS in the five different geographic locations studied was 9.6% (range: 6.7-12.4%) and varied between abattoirs (11.3%) and dairy farms (8.0%). CNS were prevalent on the carcass, milk, equipment, personnel hands, and nasal samples. Of all CNS isolates, 7.1, 10.7, 7.1, 12.5, 17.9, 10.7, 12.5, 7.1, 1.8, 5.4, 1.8, and 5.4% exhibited AMR simultaneously to single, double, 3, 4, 5, 6, 7, 7, 8, 9, 10, 11, and 13 antimicrobials, respectively. Overall, the isolates displayed 51 different AMR phenotypic patterns in which 50% of the isolates exhibited quadruple-resistance simultaneously based on the nine broad antimicrobial classes tested using 14 representative antimicrobials. The prevalence of multidrug-resistant (MDR) CNS (i.e. ≥ 3 classes of antimicrobials) was significantly (p = 0.037) different between locations with 100, 57.1, 50, 86.7, and 76.9% in Addis Ababa, Adama, Assela, Bishoftu, and Holeta, respectively. However, the prevalence of MDR CNS was not significantly (p = 0.20) different between dairy farms (87.5%) and abattoirs (71.9%). We evaluated the effect of acquiring cefoxitin-resistance of the isolates on the efficacy (i.e. inhibition zone) of the rest antimicrobials using General Linear Model after adjusting geographical locations as a random effect. Isolates with cefoxitin-resistance significantly displayed resistance to eight antimicrobials of 14 tested including amoxicillin, penicillin, cloxacillin, chloramphenicol, nalidixic acid, nitrofurantoin, and tetracycline (p = 0.000), and erythromycin (p = 0.02). On the other hand, cefoxitin-resistant isolates were susceptible to gentamicin, ciprofloxacin, kanamycin, streptomycin, and sulphamethoxazone trimethoprim (p = 0.000). Thus, antimicrobials such as gentamicin and ciprofloxacin may be an alternative therapy to treat cefoxitin-resistant CNS, as 96.4% of CNS isolates were susceptible to these antimicrobials. Overall, 94.1 and 54.5% of the CNS isolates among cefoxitin-resistant and cefoxitin-susceptible, respectively, harbored resistance to 3 or more classes of antimicrobials i.e. MDR., Conclusion: The overall prevalence of CNS in milk, meat, equipment, and food handlers in central Oromia was 9.6% but varied by location and sample source. Some specific niches such as equipment, hands, and nasal cavities of personnel are significant sites for the source of CNS. Most, but not all, MDR CNS isolates were cefoxitin-resistant. Overall, 78.6% of the CNS tested were MDR and 50% had resistance to four or more broad classes of antimicrobials. CNS in food animals (raw milk and meat), equipment, and food handlers can be the source of MDR to the public. Personnel safety and hygienic food handling practices are needed. In addition, further investigation into the risk factors for the transmission and mechanisms of resistance of the CNS is required for intervention., (© 2020 Published by Elsevier Ltd.)

Staphylococcus aureus is the major contagious bovine mastitis pathogen and has no effective vaccine. Strain variation and limited knowledge of common immunogenic antigen/s are among major constraints for developing effective vaccines. S. aureus cell surface proteins that are exposed to the host immune system constitute good vaccine candidates. The objective of this study was to compare two novel S. aureus surface protein extraction methods with biotinylation method and evaluate immune-reactivity of extracted proteins. Surface proteins were extracted from nine genetically distinct S. aureus strains from cases of bovine mastitis. After extraction, bacterial cell integrity was examined by Gram staining and electron microscopy to determine if extraction methods caused damage to cells that may release non-surface proteins. The extracted proteins were separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and evaluated for immune-reactivity using western blot. Results showed that all three extraction methods provided multiple protein bands on SDS-PAGE. Western blot result showed several immunoreactive surface proteins, in which some proteins strongly (well-resolved, thick, dark, and intense band) reacted across the nine strains tested. The three methods are valid for the extraction of surface proteins and hexadecane, and cholic acid methods are more feasible than biotinylation since both are easier, cheaper, and have minor effects on the bacterial cell. Strongly immune-reactive surface proteins may serve as potential candidates for a vaccine to control S. aureus mastitis in dairy cows., (© 2019 The Authors.)

Bovine mastitis is a significant cause of economic losses in the dairy industry. Staphylococcus aureus is one of the most common contagious mastitis pathogens, whereas Staphylococcus chromogenes increasingly became a significant cause of subclinical mastitis in dairy cows. Current mastitis control measures are not effective on all mastitis pathogens. There is no effective vaccine to control Staphylococcal mastitis in dairy cows. The objective of this study was to evaluate the immune responses and protection in dairy cows vaccinated with S. aureus surface proteins (SASP) or S. chromogenes surface proteins (SCSP). We divided eighteen Holstein dairy cows randomly into three groups of 6 animals each. We vaccinated group 1 and 2 animals with SASP and SCSP with Emulsigen-D adjuvant, respectively. We injected control (group 3) animals with PBS (pH 7.2) in Emulsigen®-D. We vaccinated animals three times at 28 and 14 days before drying off, and at dry off. Two weeks after the third vaccination, we challenged each animal by dipping all teats in S. aureus culture suspension once daily for 14 consecutive days. We evaluated milk or mammary secretion and serum antibody titers during vaccination and challenge periods. We evaluated milk samples for the number of bacteria shedding and somatic cell counts (SCC). Out of six cows vaccinated with SASP, one cow was removed from the study due to injury, two were infected clinically, another two were infected subclinically, and the remaining cow was not infected. No SCSP vaccinated cows developed clinical or subclinical mastitis. Out of six control cows, two developed clinical mastitis whereas four were infected subclinically. The SCSP vaccine cross-protected against S. aureus mastitis and reduced number of S. aureus shedding in milk. We concluded that the SCSP is a promising vaccine to control Staphylococcal mastitis in dairy cows., (Copyright © 2019 Elsevier B.V. All rights reserved.)

[This corrects the article DOI: 10.3389/fvets.2019.00102.].

Background: Peste des Petits Ruminants (PPR) is a severe, highly infectious and fatal viral disease of small ruminants. Four lineages of PPR virus have been identified globally based on sequence analysis of the nucleoprotein (N) and fusion (F) gene. The aim of this study was to isolate and genetically characterize recently circulating PPR virus in small ruminants in the eastern Amhara region in Ethiopia. A total of 28 anti-mortem samples (gum debris, nasal and ocular swab) were collected from clinically suspicious animals and examined for the presence of PPRV by a one-step RT-PCR assay. Samples positive with RT-PCR were subjected to isolation of the virus which were subsequently genetically characterized by sequencing of the nucleoprotein (N) gene and phylogenetic analysis of PPR virus (PPRV) strains., Results: Of the 28 clinical samples examined, 46.4% were positive with RT-PCR for viral nucleic acid. The PPRV was successfully isolated on CHS-20 cell line with the ovine signaling lymphocyte activation molecule (SLAM) receptor expressed on the cell surface and confirmed with RT-PCR and IFAT assay. The nucleotide sequence and phylogenetic analysis indicated that the PPRV obtained were clustered genetically with Lineage IV isolates of the virus., Conclusion: The successful isolation of the virus and molecular findings of this study confirmed active lineage IV PPRV infections among populations of sheep and goats in eastern Amhara, suggesting risks for potential spread of the disease to currently free areas. Thus, we recommend systematic vaccination to contain outbreaks in affected districts and geographically linked surrounding districts to which the disease could potentially spread due to different epidemiological linkages.

Mastitis is one of the most important diseases in dairy cows throughout the world and is responsible for significant economic losses to the dairy industry. This study was performed to characterize the genetic basis of drug resistance in Escherichia coli isolated from cases of clinical and sub-clinical bovine mastitis. A total of 224 California mastitis test (CMT)-positive milk samples were collected from December 2015 to April 2016 to characterize the phenotypic and genetic basis of antimicrobial resistance in E. coli isolated from raw milk from dairy farms found in Burayu, Sebeta, and Holeta areas of Ethiopia. The prevalence of E. coli was 7.1% (16) and both phenotypic and molecular techniques were used to identify E. coli antimicrobial susceptibility trait. The most commonly observed phenotypic resistance was against ampicillin (68.7%), sulphamethazole-trimethoprim (50%), and streptomycin (25%). Multidrug resistance phenotypes were found in 11 of 16 (68.7%) of E. coli isolates. Tetracycline (tet (A)) and chloramphenicol (cml (A)) genes were the most predominant encoding resistance genes identified (50%) each, followed by gentamycin resistance encoding gene (aac (3)-IV) (37.5%). Overall, 11 (68.7%) of the isolates had multidrug resistance genes responsible to two or more classes of antibiotics. The most common pattern detected was cml (A) and tet (A) together 37.5% followed by aac (3)-IV and tet (A) 25%. The current study indicated that raw milk could be regarded as critical source of antibiotic-resistant pathogenic E. coli.

Background: Surra is an animal trypanosomosis, caused by infection with Trypanosoma evansi and leading to severe economic loss due to mortality and morbidity. Compared to tsetse-transmitted animal trypanosomoses, little attention is given to the epidemiology and control of surra. Understanding its epidemiology is a first step in local and global efforts to control the disease. We conducted a systematic review and meta-analysis of published studies on distribution, host ranges and prevalence of T. evansi infection., Methods: Four electronic databases were searched for publications on T. evansi that met our inclusion criteria for the systematic review. Subsets of publications were subjected to meta-analysis for the pooled prevalence of T. evansi in various hosts as determined by multiple detection methods., Results: A total of 272 references published between 1906-2017 were included. Trypanosoma evansi was reported from 48 countries; largely confined to Africa and Asia with publications on natural T. evansi infections from 77% (n = 48) of countries, contrasting with seven countries in South America, and four in Europe where T. evansi is not endemic but was imported with infected animals. Although surra is a notifiable disease, many countries do not report surra cases to OIE. Trypanosoma evansi was mainly reported from dromedary camels in Africa and the Middle East, water buffaloes, cattle, dogs and horses in East and Southeast Asia. In South America, the acute form of the disease was reported in horses and dogs. Surra was also reported in a wide range of wild animals. Some rare human cases occurred in India and Vietnam. Meta-analysis on a subset of 165 publications indicated pooled prevalence of T. evansi in domestic animals ranging from 14-31%, 6-28% and 2-9% using respectively antibody detection, molecular and parasitological tests, with camels as the most affected, followed by buffalo and cattle., Conclusions: This study illustrates that T. evansi affects a wide range of domestic and wild animals in Africa, Asia and South America with highest prevalence observed in dromedary camels. For successful control of T. evansi, both locally and globally, the role of wild animals in the epidemiology of surra needs further investigation.

Camelpox is endemic in most camel rearing regions of the world, causing significant economic losses. However, its epidemiology is not extensively investigated. We conducted a cross sectional seroprevalence study of camelpox in Amibara and Awash Fentale districts in Afar region of Ethiopia from November 2014 to May 2015. In addition, participatory epidemiology (PE) was conducted to identify seasonal occurrence of the disease in the study districts. Blood samples were collected from 384 dromedary camels from 31 herds distributed in five pastoral associations (PAs) in the two districts. Serum samples were separated from the blood samples and tested for the presence of viral antibodies using virus neutralization test. Seroprevalence data were analyzed using multilevel mixed effects logistic regression models accounting for the 4-level hierarchical data structure (camels nested in herds-herds in PA, and PA in district). For the participatory data, Kendall's coefficient of concordance was used to assess agreements between the informants in identifying seasonal occurrences of the top five camel diseases. Camelpox antibodies were detected in 19.3% of camels (n = 384), 81% of herds (n = 31), and in all five PAs from the two districts in the Gabi Rasu zone of Afar region, Ethiopia. The seroprevalence did not significantly vary between herds, PAs or districts suggesting the widespread occurrence of the disease. Estimated age stratified basic reproduction number (R 0 ) was 1.25 (95% CI: 0.62-2.19). Camelpox was identified as one of the top five common camel diseases in the area. The widespread occurrence of the disease can be attributed mainly to the commingling of camels from many herds during seasonal migration in search of feed and water, a practice very common under pastoral production systems. Although the PE informants indicated the clinical disease to be more common in young animals, seropositivity was higher in older animals. Camelpox commonly occurs during the minor and major rainy seasons. In conclusion, camelpox is found to be endemic in Afar pastoral region with sporadic outbreaks occurring during rainy seasons. Vaccination and improved camel management practices particularly during the high-risk period can be viable strategies to reduce the burden of the disease., (Copyright © 2018 Elsevier B.V. All rights reserved.)

Staphylococcus aureus is a frequent and major contagious mastitis bacterial pathogen. The antibiotic treatment cure rates vary considerably from 4% to 92%. Staphylococcus aureus readily becomes resistant to antibiotics, resulting in persistent noncurable intramammary infection that usually results in culling of infected animals. Because of its notorious ability to acquire resistance to the commonly used as well as last resort antimicrobials such as methicillin and vancomycin and the development of multidrug-resistant strains, antimicrobial resistance (AMR) in S. aureus is of paramount importance in human medicine. The objective of this study was to evaluate the prevalence of AMR and genetic diversity of S. aureus isolates from milk of dairy cattle. Staphylococcus aureus isolates (n = 239) from 33 dairy farms in Tennessee were tested against 10 antimicrobials by broth microdilution method using the Sensititer system. Genetic diversity of resistant isolates was evaluated by pulsed-field gel electrophoresis (PFGE). Overall, AMR of the S. aureus isolates varied from as low as 1.3% for ceftiofur to as high as 25% for sulfadimethoxine. Out of 239 S. aureus isolates, 82 (34.3%) of them were resistant to at least 1 of the 10 antimicrobials. The AMR isolates belonged to two major PFGE types, indicating the presence of dominant clonal patterns among the resistant isolates. In general, there was a variation of prevalence of AMR within and among farms over time, with an increasing trend in tetracycline resistance. Judicious use of antimicrobials in dairy cattle farms can reduce the development of antimicrobial-resistant S. aureus.

In spite of the increasing prevalence of Streptococcus uberis mastitis, its pathogenesis and associated virulence factors are not clearly defined. The aim of this study was to identify virulence associated genes and their products that can be used to develop effective vaccine to control bovine S. uberis mastitis. S. uberis was co-cultured with primary bovine mammary epithelial cells (PBMEC) or infused into mammary gland of dairy cows. The messenger RNA (mRNA) from S. uberis associated with PBMEC after 2 h or 4 h of co-culture was purified and sequenced. Results showed that virulence-associated genes such as surface lipoprotein (slp), infection induced histidine kinase (iihK), infection induced response regulator (iirR) and extracellular sugar binding protein 1 and 2 (exsbP1 and exsbP2) were among the top-up-regulated genes. To verify this observation in vivo, quantitative real time PCR (qRT - PCR) was conducted on mRNA of S. uberis recovered from milk of infected mammary glands 24 h post infection. Results revealed that in vitro up-regulated virulence-associated genes were also significantly up regulated under in vivo conditions. The iihK and iirR are flanked by exsbP1 and exsbP2 genes and this entire operon seems to be involved in adaptation to glands micro-environment, survival and colonization of the bovine mammary glands. Based on immunogenic epitope prediction of proteins encoded by these up-regulated genes during early stages of host-bacterial interactions slp, exsbP1 and exsbP2 genes were selected, cloned and expressed in E. coli. The purified recombinant proteins (rSlP, rExsbP1 & rExsbP2) reacted strongly with convalescent serum from cows experimentally infected with S. uberis confirming that they are immunogenic. These proteins may serve as potential targets to develop an effective vaccine against S. uberis mastitis., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Background: Consumption of meat contaminated by E. coli causes a serious illness and even death to affected individuals. Recently the emerging of antibiotic resistant foodborne E. coli poses serious public health risks worldwide. However, little is known about the antibiotic resistance profile of E. coli in Ethiopia. This study aimed to determine the prevalence and Antimicrobial resistance (AMR) status of E. coli isolated from different type of meat., Methods: Overall 292 samples were collected from December 2015 to April 2016 from slaughterhouses to determine the prevalence and AMR of E. coli isolated from raw beef, mutton, chevon and chicken meat from Addis Ababa and Bishoftu, Ethiopia. The isolates were screened for AMR against commonly used antibiotics circulating in the Ethiopian market. Both phenotypic and genotypic approach were employed for AMR detection using disc diffusion test and PCR respectively., Results: The prevalence of E. coli was 63 (21.6%), indicating one sample in every five samples harbors E. coli. Among these, the highest E. coli isolates was observed in chicken meat samples (37.0%; 27), followed by mutton (23.3%; 17), chevon (20.6%; 15) and beef (5.5%; 4). Results of disk diffusion test on the 63 isolates showed that only 4.8% of them were not resistance to all antimicrobials tested. Multiple drug resistance (resistance to ≥3 drugs) was 46.0%. Significantly high resistance to ampicillin (71.4%) and tetracycline (47.6%) was observed. Identification of genes associated with AMR was also done using PCR. The prevalence of E. coli isolates harboring resistance gene responsible for tetracycline (tet(A)), beta lactams (blaCMY) and sulphanamide (sulI) antibiotics were found 65.1, 65.1 and 54.0%, respectively. Twenty-five out of the 63 (39.7% %) E. coli isolates have got antimicrobial resistance gene to three or more classes of drugs. The associations of antimicrobial resistance phenotypes and resistance genes was also determined. The detection of resistance trait against tetracycline, sulphametazole and chloramphenicol measured either phenotypically or genotypically were high., Conclusions: The rising levels of resistance E. coli to multiple antimicrobial dictate the urgent need to regulate and monitor antimicrobial use in both animals and humans.

Background: Ethiopia set an ambitious masterplan to increase chicken meat and egg production from 2015 to 2020. Poultry breeding, multiplication and distribution centers in the country have received executive order to import, amplify and distribute commercial chickens to end users. The biosecurity and the pathogen fauna of the centers have not been evaluated as to whether the centers could implement the mission effectively without any risk. Thus, the aim of this study was to evaluate the biosecurity practices and the pathogen prevalence, risk factors and their antimicrobial resistance (AMR) using Salmonella as case study., Methods: Routine farm workers of the centers were interviewed about the different management (biosecurity) practices using a checklist. Samples (n = 270) from different sources consisting of chicken's cloacal swab (n = 244), personnel hand swab (n = 9) and bedding (n = 17) were collected from three chicken multiplication centers. Standard bacteriological methods were used for the isolation of Salmonella. Disk diffusion method was used for drug sensitivity testing., Results: Antimicrobials were often over prescribed without confirming the cause of ill health and without susceptibility testing. The general biosecurity and flock management practices were substandard. Salmonella was isolated from 45 (16.7%) of the 270 samples. Its prevalence was significantly (p<0.05) associated with location of the multiplication center, 27% at Bonga and 10.6% at Hawassa. Sample type was also significantly (p<0.05) affected in that it was higher in the bedding (35.3%) and personnel hand swabs (33.3%) than in the chicken cloaca (14.8%), which demonstrates the poor biosecurity and personnel hygienic practices in the centers. All of the 45 isolates (100%) exhibited resistance to kanamycin and sulfamethoxazole-trimethoprim, nalidixic acid (97.8%), ampicillin (97.8%), cefoxitin (97.8%), streptomycin (97.8%) tetracycline (97.8%), chloramphenicol (91.3%), ciprofloxacin (31.1%), and gentamicin (0%). Alarmingly, 42 isolates (93.4%) exhibited multidrug resistance (MDR) to ≥ 8 drugs and all 45 isolates had resistance to ≥ 3 drugs. The high rate of Salmonella isolation from (i) bedding, (ii) personnel hand swabs (iii) chickens, (iv) presence of more MDR isolates, (v) coupled with poor biosecurity practices in the centers could pose a risk for spreading of pathogens and drug resistant genes to the smallholder chicken producers and the public., Conclusions: We conclude that the poultry breeding, multiplication and distribution centers in Ethiopia, as they stand currently, seem to be a source of pathogens and AMR isolates at least for Salmonella. Therefore, strict biosecurity, personnel safety, prudent drug use, regular monitoring and traceability of Salmonella serotypes or genotypes and AMR are recommended.

The aim of this study was to investigate the prevalent Bibersteinia, Mannheimia and Pasteurella serotypes, risk factors and degree of serotype co-infections in sheep and goats in the Tigray region of Ethiopia. Serum was collected from 384 sheep and goats from the Tanqua-Abergelle district of Tigray region using cross-sectional random sampling. An indirect haemagglutination test was used for serotyping. Risk factors for infections were evaluated by logistic regression. Potential clustering of multiple serotypes within individual animals due to common risk factors was evaluated by redundancy analysis. Eight serotypes were identified: all studied animals were serologically positive for at least one serotype. Overall, 355 (92·45%) of the animals were infected by four or more serotypes. Of the five risk factors studied, peasant association (PA), animal species, age (serotype A1), and bodyweight (serotype T15) were significantly associated with infection, but sex was not significant. Only PA explained a significant proportion of the variation (adjusted R 2 = 0·16) in the serological responses. After the effect of PA was accounted for, T3 and T4; A7 and Pasteurella multocida A; and A7 and T10 were positively correlated for co-infection, while T4 and T10 were less likely to be found within the same animal. Diverse serotypes were circulating in the Tigray region and could be a challenge in selecting serotypes for vaccine.

Background: Staphylococcus species cause mastitis and wound infection in livestock and food poisoning in humans through ingestion of contaminated foods, including meat and dairy products. They are evolving pathogens in that they readily acquire drug resistance, and multiple drug-resistant (MDR) isolates are increasing in human and veterinary healthcare. Therefore, this study was conducted to evaluate the prevalence of Staphylococci and their drug resistance in dairy farms and abattoir settings of Addis Ababa., Methods: In this cross-sectional study, 193 samples of milk, meat, equipment and humans working in the dairy farms and abattoir were collected (dairy farms = 72 and abattoir sources = 121). Staphylococcus isolation and identification at the species level was done according to ISO-6888-3 using biochemical characteristics. An antimicrobial susceptibility test was conducted for 43 of the isolates using 15 antimicrobial agents commonly used for humans and livestock by the Kirby Bauer disk diffusion method following CLSI guidelines., Results: Staphylococcus organism were isolated from 92 (47.7%) of the total 193 samples, 50% in the dairy farms and 46.3% in the abattoir. The isolated species were S. aureus (n = 31; 16.1%), S. intermedius (n = 21; 10.9%), S. hyicus (n = 16; 8.3%), and coagulase negative Staphylococcus (CNS) (n = 24; 12.4%). Gentamycin was effective drug as all isolates (n = 43; 100%) were susceptible to it and followed by kanamycin (n = 39; 90.7%). However, the majority of the isolates showed resistance to penicillin-G (95.3%), nalidixic acid (88.4%), cloxacillin (79.1%), vancomycin (65.1%) and cefoxitin (55.8%). Of the 15 S. aureus tested for drug susceptibility, 73.3% of them were phenotypically resistant to vancomycin (VRSA) and all of the 15 isolates showed multi-drug resistance (MDR) to >3 drugs. Also, all of the tested CNS (100%), S. hyicus (100%) and the majority of S. intermedius isolates (88.9%) developed MDR., Conclusion: Alarmingly, the Staphylococcus isolates circulating in the dairy farms and abattoir in the study area harbor MDR. High level of Staphylococcus species isolation from personnel and equipment besides food (meat and milk) samples in dairy farms and abattoir settings reveals that the hygiene practice in the dairy farm and abattoir is substandard. Prudent drug use and improved hygienic practice is recommended in the dairy farms and abattoir to safeguard the public from the risk of acquiring infections and MDR pathogenic Staphylococcus.

Background: The optimisation of trypanosomosis control programs warrants a good knowledge of the main vector of animal and human trypanosomes in sub-Saharan Africa, the tsetse fly. An important aspect of the tsetse fly population is its trypanosome infection prevalence, as it determines the intensity of the transmission of the parasite by the vector. We therefore conducted a systematic review of published studies documenting trypanosome infection prevalence from field surveys or from laboratory experiments under controlled conditions. Publications were screened in the Web of Science, PubMed and Google Scholar databases. Using the four-stage (identification, screening, eligibility and inclusion) process in the PRISMA statement the initial screened total of 605 studies were reduced to 72 studies. The microscopic examination of dissected flies (dissection method) remains the most used method to detect trypanosomes and thus constituted the main focus of this analysis. Meta-regression was performed to identify factors responsible for high trypanosome prevalence in the vectors and a random effects meta-analysis was used to report the sensitivity of molecular and serological tests using the dissection method as gold standard., Results: The overall pooled prevalence was 10.3% (95% confidence interval [CI] = 8.1%, 12.4%) and 31.0% (95% CI = 20.0%, 42.0%) for the field survey and laboratory experiment data respectively. The country and the year of publication were found to be significantly factors associated with the prevalence of trypanosome infection in tsetse flies. The alternative diagnostic tools applied to dissection positive samples were characterised by low sensitivity, and no information on the specificity was available at all., Conclusion: Both temporal and spatial variation in trypanosome infection prevalence of field collected tsetse flies exists, but further investigation on real risk factors is needed how this variation can be explained. Improving the sensitivity and determining the specificity of these alternative diagnostic tools should be a priority and will allow to estimate the prevalence of trypanosome infection in tsetse flies in high-throughput.

Background: Conventional Newcastle disease (ND) vaccination strategies in village chicken production settings is impractical due to shortage of cold-chain, unsuitability of vaccine administration routes and demanding trained personnel and hence affected its adoption. Results from earlier works elsewhere showed that the heat stable vaccines such as NDI2 are thought to be promising for village chickens. This study investigated the suitability and efficacy of Ethiopian cereal grains as carriers for the orally administrated NDI2 vaccine in chickens., Results: Of the 15 treatment groups, drinking water, cracked maize and parboiled barley induced significantly higher HI antibody titer than the other carrier grains and naive control. The higher mean HI titer of chickens in drinking-water, cracked maize and parboiled barley group resulted in 100 % survival rate. In general, there was an inverse relationship between chicken mortality (%) and mean HI titer. Chickens with higher HI antibody titers had better survival rate to the challenge experiment. Booster vaccination at age of day 35 and 105 induced progressively higher HI antibodies titers in all treatment groups., Conclusions: Vaccine coated parboiled grains could be a good carrier followed by cracked grains while untreated vaccine carrier grains had lower serological responses and protection levels. The current finding gives insights on suitable vaccine delivery system in villages with weak health and transportation infrastructure, unreliable electricity, and minimally trained health workers without catching chickens individually.