Multidrug-Resistant Staphylococcus aureus Strains Thrive in Dairy and Beef Production, Processing, and Supply Lines in Five Geographical Areas in Ethiopia.

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.

Fecal Microbiota Transplantation Reduces Campylobacter jejuni Colonization in Young Broiler Chickens Challenged by Oral Gavage but Not by Seeder Birds.

Campylobacter spp., particularly C. jejuni and C. coli, are major food safety concerns, transmitted to humans mainly via contaminated poultry meat. In a previous study, we found that some commercial broiler farms consistently produced Campylobacter-free flocks while others consistently reared Campylobacter-colonized flocks, and significant differences in the gut microbiota compositions between the two types of farm categories were revealed. Therefore, we hypothesized that gut microbiota influences Campylobacter colonization in poultry and that the microbiota from Campylobacter-free flocks may confer colonization resistance to Campylobacter in the chicken intestine. In this study, two fecal microbiota transplantation (FMT) trials were performed to test the hypothesis. Newly hatched chicks were given FMT via oral gavage of the cecal content of Campylobacter-free adult chickens (treatment groups) or PBS (control groups) before the feed consumption. Approximately two weeks after the FMT, the birds were challenged with C. jejuni either by oral gavage (trial 1) or by co-mingling with Campylobacter-colonized seeder birds (trial 2) to evaluate the potential protective effect of the FMT. Cecal contents were collected (3 times, 5 days apart) to determine the Campylobacter colonization levels via culture and microbiota compositions via 16S rRNA gene sequencing. FMT reduced cecal Campylobacter colonization significantly (log10 1.2-2.54 CFU/g) in trial 1 but not in trial 2, although FMT significantly impacted the diversity and compositions of the gut microbiota in both trials. Several genera, such as Butyricimonas, Parabacteroides, Parasutterella, Bilophila, Fournierella, Phascolarctobacterium, and Helicobacter, had increased abundance in the FMT-treated groups in both trials. Furthermore, Campylobacter abundance was found to be negatively correlated with the Escherichia and Ruminococcus_torques_group genera. These findings indicate that even though FMT with adult cecal microbiota can positively affect the subsequent development of the gut microbiota in young broilers, its inhibitory effect on Campylobacter colonization varies and appears to be influenced by the challenge models.

Efficacy of Penicillin-Streptomycin Brands against Staphylococcus aureus: Concordance between Veterinary Clinicians' Perception and the Realities.

Antibiotics must be safe and effective for use in both human and veterinary medicine. However, information about the efficacy of different brands of antibiotics commonly used in veterinary practices is lacking in Ethiopia. In this study, we determined the efficacy of three brands of penicillin-streptomycin (Pen&strep, Penstrep, and Pro&strep) by performing antimicrobial susceptibility testing against Staphylococcus aureus isolated from cow milk from dairy farms in the towns of Sebata and Bishoftu, Central Ethiopia. We also assessed the knowledge, attitudes, and practices (KAP) of veterinarians regarding the quality and use of brand antibiotics and the antibiotic utilization practices of dairy farm personnel using a structured questionnaire. Of 43 S. aureus isolated and tested, 33 (77%), 10 (23%), and 1 (2%) were susceptible to brands A, B, and C, respectively. According to the respondents, all of them reported that penstrep is the most prescribed antibiotic in dairy farms (100%), followed by oxytetracycline (78%) and sulfa drugs (52%). All veterinarians perceived that antibiotics imported from Western countries have a higher efficacy than those from Eastern countries, and they preferred brand A to the other brands, witnessing its better clinical outcome. The majority (87%) and a little more than half (53%) of the respondents perceived the overuse of antibiotics in veterinary clinics and dairy farms, respectively. Our study revealed the better efficacy of brand A against S. aureus compared to the other brands. Interestingly, the veterinarians' perception of and preference toward the use of brand antibiotics agreed with the findings of our antibacterial susceptibility testing. The prudent use of brand A is critically important for sustaining effective treatment, avoiding the risk of antimicrobial resistance, and helping to address animal welfare issues.

Influence of Single Dose Enrofloxacin Injection on Development of Fluoroquinolone Resistance in Campylobacter jejuni in Calves.

Fluoroquinolone (FQ) resistance in a major foodborne bacterial pathogen, Campylobacter jejuni, derived from cattle has recently become prevalent and poses a significant public health concern. However, the underlying factors for this increase are not entirely clear. To evaluate the effect of enrofloxacin treatment on FQ-resistance development in C. jejuni, 35 commercial calves were equally divided into five groups (Groups 1-5) and were orally inoculated with FQ-susceptible (FQ-S) C. jejuni. Eight days later, Groups 4 and 5 were challenged with Mannheimia haemolytica via a transtracheal route to induce a respiratory disease; after 8 days, Groups 2, 3, 4, and 5 were injected subcutaneously with enrofloxacin (7.5 mg/kg for Groups 2 and 4, and 12.5 mg/kg for Groups 3 and 5). Colonization levels by FQ-resistant (FQ-R) and FQ-S Campylobacter in rectal feces were determined via differential culture throughout the experiment. Before oral inoculation with C. jejuni, only five calves were naturally colonized by Campylobacter, four of which were also colonized by FQ-R C. jejuni (three in Group 1 and one in Group 3). Soon after the oral inoculation, almost all calves in the groups became stably colonized by FQ-S C. jejuni (~3-6 log10 CFU/g), except that the four calves that were pre-colonized before inoculation remained positive with both FQ-R and FQ-S C. jejuni. Following enrofloxacin administration, C. jejuni colonization declined sharply and rapidly in all treated groups to undetectable levels; however, the vast majority of the animals were recolonized by C. jejuni at comparable levels 72 h after the treatment. Notably, no FQ-R C. jejuni was detected in any of the calves that received enrofloxacin, regardless of the drug dose used or disease status of the animals. The lack of detection of FQ-R C. jejuni was likely due to the localized high concentration of the antibiotic in the intestine, which may have prevented the emergence of the FQ-R mutant. These findings indicate that single-dose enrofloxacin use in cattle poses a low risk for selection of de novo FQ-R mutants in C. jejuni.

Impacts of Gut Microbiota on the Immune System and Fecal Microbiota Transplantation as a Re-Emerging Therapy for Autoimmune Diseases.

The enormous and diverse population of microorganisms residing in the digestive tracts of humans and animals influence the development, regulation, and function of the immune system. Recently, the understanding of the association between autoimmune diseases and gut microbiota has been improved due to the innovation of high-throughput sequencing technologies with high resolutions. Several studies have reported perturbation of gut microbiota as one of the factors playing a role in the pathogenesis of many diseases, such as inflammatory bowel disease, recurrent diarrhea due to Clostridioides difficile infections. Restoration of healthy gut microbiota by transferring fecal material from a healthy donor to a sick recipient, called fecal microbiota transplantation (FMT), has resolved or improved symptoms of autoimmune diseases. This (re)emerging therapy was approved for the treatment of drug-resistant recurrent C. difficile infections in 2013 by the U.S. Food and Drug Administration. Numerous human and animal studies have demonstrated FMT has the potential as the next generation therapy to control autoimmune and other health problems. Alas, this new therapeutic method has limitations, including the risk of transferring antibiotic-resistant pathogens or transmission of genes from donors to recipients and/or exacerbating the conditions in some patients. Therefore, continued research is needed to elucidate the mechanisms by which gut microbiota is involved in the pathogenesis of autoimmune diseases and to improve the efficacy and optimize the preparation of FMT for different disease conditions, and to tailor FMT to meet the needs in both humans and animals. The prospect of FMT therapy includes shifting from the current practice of using the whole fecal materials to the more aesthetic transfer of selective microbial consortia assembled in vitro or using their metabolic products.

Antimicrobial Resistance of Staphylococci at Animal Human Interface in Smallholders and Dairy Farms in Central Oromia, Ethiopia.

Purpose: Staphylococcus species come from a variety of sources and can contaminate milk during milking, cause mastitis and other diseases in animals and humans. The enterotoxins they produce cause food poisoning. Our objectives were to isolate, biochemically characterize, and determine antimicrobial susceptibility profiles of Staphylococcus species from dairy farms in central Oromia, Ethiopia. Methods: A total of 339 samples (n = 135 [raw milk], n = 135 [udders' swabs], n = 25 [milkers' hands swabs], n = 44 [pooled milking utensils' swabs]) were collected from smallholders and dairy farms. Bacteriological culture and biochemical tests were performed to isolate and identify Staphylococcus species, and the Kirby Bauer disk diffusion method was used for antimicrobial susceptibility testing. Results: Across all sample types and dairy farms, 247 (72.9%) Staphylococcus isolates were obtained which comprised of 101 (74.8%) isolates from raw milk, 98 (72.6%) from udder swabs, 30 (68.2%) from pooled utensil swabs, and 18 (72%) from milkers' hand swabs. Fifty coagulase-positive Staphylococcus isolates (20 S. aureus, 20 S. hyicus and 10 S. intermedius) subjected to antimicrobial susceptibility tests have shown various degrees of resistance. All S. aureus isolates were 100% resistant to ampicillin and penicillin. Out of 20 S. hyicus isolates, 90% were resistant to ampicillin and 85% to penicillin. S. intermedius isolates (n=10) were 70% resistant to nalidixic acid and penicillin whilst remaining 100% resistant to ampicillin. Five S. aureus, three S. intermedius and two S. hyicus isolates from raw milk, milk utensil swabs and milkers' hand swabs were multidrug-resistant (resistance to at least three classes of antimicrobials). Conclusion: This study revealed a high prevalence of staphylococci in the dairy cattle, milkers and milking utensils with multidrug-resistant coagulase-positive Staphylococcus species suggesting the significance of pasteurization. Further research is encouraged on the factors leading to antibiotic resistance in Staphylococcus species.

Effect of Danofloxacin Treatment on the Development of Fluoroquinolone Resistance in Campylobacter jejuni in Calves.

Campylobacter is a leading cause of foodborne gastroenteritis. Recent studies have indicated a rise in fluoroquinolone-resistant (FQ-R) Campylobacter in cattle, where FQ is used to control bovine respiratory disease (BRD). To assess the effect of danofloxacin treatment on the development of FQ-resistance in C. jejuni, 30 commercial calves were divided into Group 1, Group 2, and Group 3 (n = 10), and were all inoculated orally with FQ-susceptible (FQ-S) C. jejuni; seven days later, Group 3 was challenged with transtracheal Mannheimia haemolytica, and one week later, Group 2 and Group 3 were injected subcutaneously with danofloxacin. Rectal feces were collected to determine relative percentages of FQ-R Campylobacter via culture. Before oral inoculation with C. jejuni, 87% of calves were naturally colonized by FQ-R C. jejuni. Two days after the inoculation, FQ-R C. jejuni decreased substantially in the majority of calves. Within 24 h of danofloxacin injection, almost all C. jejuni populations shifted to an FQ-R phenotype in both FQ-treated groups, which was only transitory, as FQ-S strains became predominant during later periods. Genotyping indicated that the spike seen in FQ-R C. jejuni populations following the injection was due mainly to enrichment of preexisting FQ-R C. jejuni, rather than development of de novo FQ resistance in susceptible strains. These results provide important insights into the dynamic changes of FQ-resistant Campylobacter in cattle in response to FQ treatment.

Comparisons of plasma and fecal pharmacokinetics of danofloxacin and enrofloxacin in healthy and Mannheimia haemolytica infected calves.

Danofloxacin and enrofloxacin are fluoroquinolones (FQs) used to treat and control bovine respiratory disease (BRD) complex. While low toxicity, high bactericidal activity, and availability in single and multiple dosing regimens make them preferable, the increasing incidence of FQ-resistance in foodborne pathogens and effects on gut microbiota necessitate evaluating their pharmacokinetics (PKs). The objective of this study was to determine the exposure level of gut microbiota to subcutaneously administered FQs and compare their PKs between plasma and feces in healthy and Mannheimia haemolytica infected calves. A single dose of danofloxacin (8 mg/kg), low dose (7.5 mg/kg), or high dose (12.5 mg/kg) of enrofloxacin was administered to calves. Blood and feces were collected from calves under experimental conditions over 48 h, and FQ concentrations were measured using Ultra High-Pressure Liquid Chromatography. While moderate BRD signs were exhibited in most calves in the infected cohorts, the plasma PKs were similar between healthy and sick calves. However, the fecal danofloxacin concentration was lower in the BRD group (area under concentration-time curve [AUCinf], BRD median = 2627, healthy median = 2941 h*µg/mL, adj.P = 0.005). The dose normalized plasma and fecal danofloxacin concentrations were higher than those of enrofloxacin and its metabolite ciprofloxacin. Further, FQs had several fold higher overall concentrations in feces than in plasma in both groups. In conclusion, parenterally administered FQs expose gut microbiota to high concentrations of the antibiotics.

Veterinary Drug Prescribing Practices at Selected District Veterinary Clinics of Rift Valley Areas of Ethiopia.

The rational use of drugs in veterinary medicine has various significances, such as reducing the risk of drug resistance, increasing efficacy, reducing drug residue, and decreasing adverse drug reactions. A retrospective study was conducted to assess veterinary drug prescribing practices at Batu and Arsi-Negelle district veterinary clinics in the rift valley areas of Ethiopia. A total of 2,464 cases were recorded from the case registration books at both the clinics for diseases treated between September 2012 and February 2015. The study results showed that for a total of 2,464 cases diagnosed at both clinics, 3,811 different drugs were prescribed, with an average per encounter of 1.6. Among the total drugs, oxytetracycline, ivermectin, penstrep, sulfa drugs, and albendazole were the most leading prescribed drugs with a frequency of 43.0%, 17.6%, 10.2%, 6.5%, and 1.3%, respectively. All drugs were prescribed by the generic name without any laboratory support of the disease. About 68.3% of the cases were diagnosed by unspecified professionals, whereas 21.7% and 10.1% were done by animal health assistants and veterinarians, respectively. The prescribing practices showed 61.0% of antibiotics and 29.7% of anthelmintics where 45.3% and 54.7% of antibiotics and 17.8% and 82.2% of anthelmintics were given at Batu and Arsi-Negelle veterinary clinics, respectively. Of the prescribed drugs, 4.6% oxytetracycline and 2.6% penstrep were prescribed irrationally to treat diseases that were tentatively diagnosed as parasitic cases. Similarly, 40.5% ivermectin and 17.7% albendazole were prescribed for bacterial infections. In conclusion, this study revealed problems in antibiotics and anthelmintics use, description of routes of administration and length of treatment, and shortage of laboratory diagnostic facilities. Therefore, veterinary drugs, particularly antibiotics and anthelmintics, should be used appropriately to safeguard the public from residual drug impacts and resistance development.

Danofloxacin Treatment Alters the Diversity and Resistome Profile of Gut Microbiota in Calves.

Fluoroquinolones, such as danofloxacin, are used to control bovine respiratory disease complex in beef cattle; however, little is known about their effects on gut microbiota and resistome. The objectives were to evaluate the effect of subcutaneously administered danofloxacin on gut microbiota and resistome, and the composition of Campylobacter in calves. Twenty calves were injected with a single dose of danofloxacin, and ten calves were kept as a control. The effects of danofloxacin on microbiota and the resistome were assessed using 16S rRNA sequencing, quantitative real-time PCR, and metagenomic Hi-C ProxiMeta. Alpha and beta diversities were significantly different (p < 0.05) between pre-and post-treatment samples, and the compositions of several bacterial taxa shifted. The patterns of association between the compositions of Campylobacter and other genera were affected by danofloxacin. Antimicrobial resistance genes (ARGs) conferring resistance to five antibiotics were identified with their respective reservoirs. Following the treatment, some ARGs (e.g., ant9, tet40, tetW) increased in frequencies and host ranges, suggesting initiation of horizontal gene transfer, and new ARGs (aac6, ermF, tetL, tetX) were detected in the post-treatment samples. In conclusion, danofloxacin induced alterations of gut microbiota and selection and enrichment of resistance genes even against antibiotics that are unrelated to danofloxacin.

Enrofloxacin Alters Fecal Microbiota and Resistome Irrespective of Its Dose in Calves.

Enrofloxacin is a fluoroquinolone drug used to prevent and control bovine respiratory disease (BRD) complex in multiple or single doses, ranging from 7.5 to 12.5 mg/kg body weight. Here, we examined the effects of high and low doses of a single subcutaneously injected enrofloxacin on gut microbiota and resistome in calves. Thirty-five calves sourced for this study were divided into five groups: control (n = 7), two low dose groups (n = 14, 7.5 mg/kg), and two high dose groups (n = 14, 12.5 mg/kg). One group in the low and high dose groups was challenged with Mannheimia haemolytica to induce BRD. Both alpha and beta diversities were significantly different between pre- and post-treatment microbial communities (q < 0.05). The high dose caused a shift in a larger number of genera than the low dose. Using metagenomic ProxiMeta Hi-C, 32 unique antimicrobial resistance genes (ARGs) conferring resistance to six antibiotic classes were detected with their reservoirs, and the high dose favored clonal expansion of ARG-carrying bacterial hosts. In conclusion, enrofloxacin treatment can alter fecal microbiota and resistome irrespective of its dose. Hi-C sequencing provides significant benefits for unlocking new insights into the ARG ecology of complex samples; however, limitations in sample size and sequencing depth suggest that further work is required to validate the findings.

Pharmacokinetics of tulathromycin in pregnant ewes (Ovis aries) challenged with Campylobacter jejuni.

The purpose of this study was to evaluate the pharmacokinetics of tulathromycin in the plasma and maternal and fetal tissues of pregnant ewes when administered within 24 hours of a single, IV Campylobacter jejuni (C. jejuni) challenge. Twelve, pregnant ewes between 72-92 days of gestation were challenged IV with C. jejuni IA3902 and then treated with 1.1 ml/45.36 kg of tulathromycin subcutaneously 18 hours post-challenge. Ewes were bled at predetermined time points and euthanized either at a predetermined time point or following the observation of vaginal bleeding or abortion. Following euthanasia, tissues were collected for bacterial culture, pharmacokinetics and histologic examination. The maximum (geometric) mean tulathromycin plasma concentration was estimated at 0.302 µg/mL, with a peak level observed at around 1.2 hours. The apparent systemic clearance of tulathromycin was estimated at 16.6 L/h (or 0.28 L/kg/h) with an elimination half-life estimated at approximately 22 hours. The mean tissue concentrations were highest in the uterus (2.464 µg/g) and placentome (0.484 µg/g), and were lowest in fetal liver (0.11 µg/g) and fetal lung (0.03 µg/g). Compared to previous reports, results of this study demonstrate that prior IV administration of C. jejuni appeared to substantially alter the pharmacokinetics of tulathromycin, reducing both the peak plasma concentrations and elimination half-life. However, additional controlled trials are required to confirm those observations.

Tracking Reservoirs of Antimicrobial Resistance Genes in a Complex Microbial Community Using Metagenomic Hi-C: The Case of Bovine Digital Dermatitis.

Bovine digital dermatitis (DD) is a contagious infectious cause of lameness in cattle with unknown definitive etiologies. Many of the bacterial species detected in metagenomic analyses of DD lesions are difficult to culture, and their antimicrobial resistance status is largely unknown. Recently, a novel proximity ligation-guided metagenomic approach (Hi-C ProxiMeta) has been used to identify bacterial reservoirs of antimicrobial resistance genes (ARGs) directly from microbial communities, without the need to culture individual bacteria. The objective of this study was to track tetracycline resistance determinants in bacteria involved in DD pathogenesis using Hi-C. A pooled sample of macerated tissues from clinical DD lesions was used for this purpose. Metagenome deconvolution using ProxiMeta resulted in the creation of 40 metagenome-assembled genomes with ≥80% complete genomes, classified into five phyla. Further, 1959 tetracycline resistance genes and ARGs conferring resistance to aminoglycoside, beta-lactams, sulfonamide, phenicol, lincosamide, and erythromycin were identified along with their bacterial hosts. In conclusion, the widespread distribution of genes conferring resistance against tetracycline and other antimicrobials in bacteria of DD lesions is reported for the first time. Use of proximity ligation to identify microorganisms hosting specific ARGs holds promise for tracking ARGs transmission in complex microbial communities.

A Homologous Bacterin Protects Sheep against Abortion Induced by a Hypervirulent Campylobacter jejuni Clone.

Campylobacter jejuni clone SA has emerged as the predominant cause of ovine abortion outbreaks in the United States (US). Despite the fact that commercial Campylobacter vaccines are available, their efficacy in protecting abortion induced by C. jejuni clone SA is uncertain, and a protective vaccine is needed to control the disease. In this study, an experimental homologous bacterin (made of a clone SA isolate) and two commercial Campylobacter vaccines were evaluated for their protection against C. jejuni clone SA-induced sheep abortion. All vaccines induced high levels of antibodies against C. jejuni clone SA in pregnant ewes, but only the experimental homologous bacterin produced significant protection (80%). Immunoblotting showed that the experimental vaccine elicited more specific antibodies against C. jejuni clone SA. These findings strongly suggest the necessity of developing a homologous vaccine for the control C. jejuni clone SA induced abortion on sheep farms.

Distribution and antimicrobial resistance profile of coagulase-negative staphylococci from cattle, equipment, and personnel on dairy farm and abattoir settings.

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.

Experimental evaluation of tulathromycin as a treatment for Campylobacter jejuni abortion in pregnant ewes.

OBJECTIVE: To evaluate the efficacy of tulathromycin for prevention of abortion in pregnant ewes when administered within 24 hours after experimental inoculation with Campylobacter jejuni. ANIMALS: 20 pregnant ewes between 72 and 92 days of gestation. PROCEDURES: All ewes were inoculated with a field strain of C jejuni (8.5 × 108 to 10.6 × 108 CFUs, IV). Eighteen hours later, ewes received either tulathromycin (1.1 mL/45 kg [2.4 mg/kg], SC; n = 10) or sterile saline (0.9% NaCl) solution (1.1 mL/45 kg, SC; sham; 10). Ewes were euthanized immediately after observation of vaginal bleeding, abortion, or completion of a 21-day observation period. Necropsy was performed on all ewes, and tissue specimens were obtained for bacterial culture and histologic examination. RESULTS: 1 sham-treated ewe and 1 tulathromycin-treated ewe developed signs of severe endotoxemia and were euthanized within 24 hours after C jejuni inoculation. Seven sham-treated and 2 tulathromycin-treated ewes developed vaginal bleeding or aborted and were euthanized between 4 and 21 days after C jejuni inoculation. The proportion of tulathromycin-treated ewes that developed vaginal bleeding or aborted during the 21 days after C jejuni inoculation (2/9) was significantly less than that for the sham-treated ewes (7/9). CONCLUSIONS AND CLINICAL RELEVANCE: Results suggested that administration of tulathromycin to pregnant ewes following exposure to C jejuni was effective in decreasing the number of C jejuni-induced abortions. Because of concerns regarding the development of macrolide resistance among Campylobacter strains, prophylactic use of tulathromycin in sheep is not recommended.

Veterinary medicinal product usage among food animal producers and its health implications in Central Ethiopia.

BACKGROUND: Antimicrobials and anthelmintics are the most commonly used veterinary drugs to control animal diseases. However, widespread use of these drugs could contribute to the emergence of drug resistance. Information on the practice of antimicrobial usage among food animal raising communities in Central Ethiopia is scarce. We used a standardised questionnaire survey to assess knowledge, awareness, and practices related to drug use and resistance in food animals among the farmers in and around Bishoftu town. RESULTS: Of the total of 220 livestock owners interviewed, around 80% of the respondents were not able to define what antimicrobials are and for what purposes they are used. Only 14.1% (n = 31) of the respondents had awareness about antimicrobial resistance (AMR) and its consequences; and 35.5% (n = 11/31) and 9.7% (n = 3/31) of them agreed that the irrational use of antimicrobials in animals could lead to AMR in animals and humans. Oxytetracycline was the most commonly available antibiotic in veterinary drug shops/pharmacies and the most widely used drug in the area. However, 43.3% of the respondents did not see clinical improvements after using antibiotics. Similarly, the respondents explained that no response was observed in 73.3, 70.8 and 52.5% of the cases after medication with anthelmintics, antiprotozoal and acaricides, respectively. About 56.7% of the respondents considered traditional medicines equally important to modern medicines. It was also noted that there were illegal drug vendors, dispensing medicines under unfavourable conditions which include a direct exposure to sunlight, which practice violates the drug handling and storage recommendations given by WHO. CONCLUSION: The study revealed that there is a general lack of awareness among food animal owners about the correct use of antibiotics and anthelmintics. The widespread misuse and improper drug dispensing and handling practices observed in this study can affect the drug quality and can also contribute to the development of drug resistance in central Ethiopia.

Risk Perceptions and Protective Behaviors Toward Bovine Tuberculosis Among Abattoir and Butcher Workers in Ethiopia.

Bovine Tuberculosis (BTB) is a serious cause of economic losses and public health threat, especially in developing countries. Humans acquire BTB through consumption of raw or undercooked meat, inhalation of aerosol and occupational exposure. A cross-disciplinary approach to study diseases connecting society and biology helps to understand the ways in which social, cultural, behavioral, and economic circumstances influence a healthy life. The objective of this study was to assess the risk perceptions and protective behaviors toward BTB among abattoir and butcher workers in central Ethiopia. A health belief model was used to generate the desired data following health belief model constructs. A total of 300 meat handlers working in local abattoirs, export abattoirs and butcher houses in Bishoftu, Modjo, Dukem, and Akaki towns of central Ethiopia were selected using a systematic random sampling method. Univariate and multivariable logistic regression analysis were used to assess factors associated with risk of exposure to BTB through the consumption of raw meat. The results showed that among the study participants, 95% heard about BTB and 93% knew that eating raw meat could be a source of BTB for humans. More than 62.7% of the respondents in the high risk group strongly agreed that contracting BTB would prevent them from coming to work, keep them in bed for an extended period of time and cause death. The majority of the respondents believed that free provision of personal protective clothing, compensation with test and slaughter campaigns, television and radio advertisements, educational programs and government-imposed penalties would help in prevention of BTB. Despite the high perceived severity and risk perception, the multivarable logistic regression model showed low-risk protective behavior among male (OR: 2.3, 95% CI: 1.2-4.3) and older age (>30) individuals (OR: 14.4 95% CI: 2.1-125.8). The study also noted the importance of media for health education as means for prevention of BTB. The authors strongly recommended the need of promotion of behavioral change toward the consumption of raw meat wich would have potential implications for the public health impacts of zoonotic tuberculosis and ultimately help national and global efforts toward prevention and control of tuberculosis.

A Smartphone-Based Application Improves the Accuracy, Completeness, and Timeliness of Cattle Disease Reporting and Surveillance in Ethiopia.

Accurate disease reporting, ideally in near real time, is a prerequisite to detecting disease outbreaks and implementing appropriate measures for their control. This study compared the performance of the traditional paper-based approach to animal disease reporting in Ethiopia to one using an application running on smartphones. In the traditional approach, the total number of cases for each disease or syndrome was aggregated by animal species and reported to each administrative level at monthly intervals; while in the case of the smartphone application demographic information, a detailed list of presenting signs, in addition to the putative disease diagnosis were immediately available to all administrative levels via a Cloud-based server. While the smartphone-based approach resulted in much more timely reporting, there were delays due to limited connectivity; these ranged on average from 2 days (in well-connected areas) up to 13 days (in more rural locations). We outline the challenges that would likely be associated with any widespread rollout of a smartphone-based approach such as the one described in this study but demonstrate that in the long run the approach offers significant benefits in terms of timeliness of disease reporting, improved data integrity and greatly improved animal disease surveillance.

Assisting differential clinical diagnosis of cattle diseases using smartphone-based technology in low resource settings: a pilot study.

BACKGROUND: The recent rise in mobile phone use and increased signal coverage has created opportunities for growth of the mobile Health sector in many low resource settings. This pilot study explores the use of a smartphone-based application, VetAfrica-Ethiopia, in assisting diagnosis of cattle diseases. We used a modified Delphi protocol to select important diseases and Bayesian algorithms to estimate the related disease probabilities based on various clinical signs being present in Ethiopian cattle. RESULTS: A total of 928 cases were diagnosed during the study period across three regions of Ethiopia, around 70% of which were covered by diseases included in VetAfrica-Ethiopia. Parasitic Gastroenteritis (26%), Blackleg (8.5%), Fasciolosis (8.4%), Pasteurellosis (7.4%), Colibacillosis (6.4%), Lumpy skin disease (5.5%) and CBPP (5.0%) were the most commonly occurring diseases. The highest (84%) and lowest (30%) levels of matching between diagnoses made by student practitioners and VetAfrica-Ethiopia were for Babesiosis and Pasteurellosis, respectively. Multiple-variable logistic regression analysis indicated that the putative disease indicated, the practitioner involved, and the level of confidence associated with the prediction made by VetAfrica-Ethiopia were major determinants of the likelihood that a diagnostic match would be obtained. CONCLUSIONS: This pilot study demonstrated that the use of such applications can be a valuable means of assisting less experienced animal health professionals in carrying out disease diagnosis which may lead to increased animal productivity through appropriate treatment.