Occurrence, antimicrobial susceptibility, and resistance genes of Staphylococcus aureus in milk and milk products in the Arsi highlands of Ethiopia.

BACKGROUND: In Ethiopia, milk production and handling practices often lack proper hygiene measures, leading to the potential contamination of milk and milk products with Staphylococcus aureus (S. aureus), including methicillin-resistant strains, posing significant public health concerns. This study aimed to investigate the occurrence, antimicrobial susceptibility profiles and presence of resistance genes in S. aureus strains isolated from milk and milk products. METHODS: A cross-sectional study was conducted in the Arsi highlands, Oromia, Ethiopia from March 2022 to February 2023. A total of 503 milk and milk product samples were collected, comprising 259 raw milk, 219 cottage cheese, and 25 traditional yogurt samples. S. aureus isolation and coagulase-positive staphylococci enumeration were performed using Baird-Parker agar supplemented with tellurite and egg yolk. S. aureus was further characterized based on colony morphology, Gram stain, mannitol fermentation, catalase test, and coagulase test. Phenotypic antimicrobial resistance was assessed using the Kirby-Bauer disc diffusion method, while the polymerase chain reaction (PCR) was employed for confirming the presence of S. aureus and detecting antimicrobial resistance genes. RESULTS: S. aureus was detected in 24.9% of the milk and milk products, with the highest occurrence in raw milk (40.9%), followed by yogurt (20%), and cottage cheese (6.4%). The geometric mean for coagulase-positive staphylococci counts in raw milk, yogurt, and cottage cheese was 4.6, 3.8, and 3.2 log10 CFU/mL, respectively. Antimicrobial resistance analysis revealed high levels of resistance to ampicillin (89.7%) and penicillin G (87.2%), with 71.8% of the isolates demonstrating multidrug resistance. Of the 16 S. aureus isolates analyzed using PCR, all were found to carry the nuc gene, with the mecA and blaZ genes detected in 50% of these isolates each. CONCLUSION: This study revealed the widespread distribution of S. aureus in milk and milk products in the Arsi highlands of Ethiopia. The isolates displayed high resistance to ampicillin and penicillin, with a concerning level of multidrug resistance. The detection of the mecA and blaZ genes in selected isolates is of particular concern, highlighting a potential public health hazard and posing a challenge to effective antimicrobial treatment. These findings highlight the urgent need to enhance hygiene standards in milk and milk product handling and promote the rational use of antimicrobial drugs. Provision of adequate training for all individuals involved in the dairy sector can help minimize contamination. These measures are crucial in addressing the threats posed by S. aureus, including methicillin-resistant strains, and ensuring the safety of milk and its products for consumers.

Isolation, antimicrobial susceptibility patterns, and risk factors assessment of non-typhoidal Salmonella from apparently healthy and diarrheic dogs.

BACKGROUND: Dogs are one of the important asymptomatic carriers of antimicrobial resistant and potentially pathogenic strains of Salmonella. They can harbor large bacterial load in the intestines and mesenteric lymph nodes which can be shed in their feces with the possibility of transmission to humans. Therefore, a cross-sectional study was conducted with the objectives of estimating the prevalence of non-typhoidal Salmonella, assessing the risk factors for dog's Salmonella carriage, and profiling the antimicrobial resistance pattern of Salmonella isolates among housed dogs in Harar town, Eastern Ethiopia. A total of 415 rectal swab samples were collected from randomly selected dogs. Samples were examined for non-typhoidal Salmonella using standard bacteriologic culture and biochemical tests. The disk diffusion method (Kirby-Bauer test) was employed to evaluate the isolates for their susceptibility against five antimicrobials. RESULTS: Non-typhoidal Salmonella were isolated from 26 (6.3%) of the rectal swab samples, with significantly higher occurrence in diarrheic (15.2%) than non-diarrheic (5.5%) dogs. The risk of Salmonella harboring was significantly higher in female dogs than in male dogs (OR = 2.5, p = 0.027). Dogs fecal shedding of Salmonella was relatively higher in households who used offal as a main feed type for their dogs (23.1%; 95% CI = 5-53.8) than those who used leftover food (10.1%; 95% CI = 5.7-16.1) and practiced mixed feeding system (17%; 95% CI = 7.6-30.8). Salmonella isolates showed higher resistance to ampicillin (41.7%), while all isolates were fully susceptible to gentamicin. Moreover, 58.3% of Salmonella isolates showed resistance to at least one of the tested antimicrobials. Majorities (72.7%) of the dog owners had no awareness on the risk of zoonotic salmonellosis from dog and all of the respondents use bare hand to clean dog kennel. CONCLUSION: Our study reveals the importance of both diarrheic and apparently healthy housed dogs in the harboring and shedding of antimicrobial resistant non-typhoidal Salmonella. The risk of non-typhoidal Salmonella spread among pet owners is not negligible, especially in households who use offal as main feed type. Therefore, an integrated approach such as: proper dog handling practices; continuous evaluation of antimicrobial resistance; and rational use of antimicrobials in the field of veterinary sector are necessary to tackle the problem.

First-time detection of mycobacterium species from goats in Ethiopia.

Tuberculosis (TB) is an important zoonosis affecting a wide range of hosts. An abattoir study was conducted on 1,536 randomly selected male goats slaughtered at Modjo Modern Export Abattoir to determine the prevalence of tuberculosis in slaughtered goats. Carcasses and organs of all the study animals were first examined by routine meat inspection followed by detailed meat inspection. Samples from tuberculous lesions were cultured for mycobacterial isolation and identification. Histopathology was done on 31 samples with tuberculous lesions. Detailed meat inspection detected 65 (4.2%; 95% confidence interval (CI) = 3.3-5.4%) tuberculous lesions. From these, 20 (30.8%) samples were confirmed mycobacterium positive on culture, out of which 18 were Mycobacterium bovis and two were Mycobacterium tuberculosis. Routine meat inspection failed to detect tuberculous lesions in 23% of carcasses with TB lesions detected by detailed examination. However, no statistically significant difference was observed between both methods in detecting tuberculous lesions (Kappa = 0.87). Origin and age of the goats did not statistically affect the disease prevalence (P > 0.05). Histopathologic lesions were observed in 21 samples (68%; 95% CI = 50.1-81.4%) out of the 31 carcasses with gross tuberculous lesions examined by histopathology. Eighteen (58%) tuberculous samples positive for histopathology were also culture positive. The sensitivity and specificity of histopathology were 90% (95% CI = 76.9-100%) and 72.7% (95% CI = 46.4-99%), respectively, using culture as a reference test. To the best of our knowledge, this is the first report of caprine tuberculosis from Ethiopia. Further studies are required at the farm level to determine the prevalence of tuberculosis in the general goat population.

Bovine cysticercosis and human taeniasis in a rural community in Ethiopia.

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.

Dipylidium caninum Infection in Dogs and Humans in Bishoftu Town, Ethiopia.

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.

Zoonotic metacestodes and associated financial loss from cattle slaughtered at Yabello municipal abattoir, Borana-Oromia, Ethiopia.

Cross-sectional study was carried out to determine the prevalence and estimate financial losses associated with zoonotic metacestodes in cattle originated from Borana Pastoral Farming System and slaughtered at Yabello Municipal Abattoir, Ethiopia. A total of 384 cattle were randomly selected at the abattoir. Detailed postmortem examination, cyst count and fertile test were conducted. Significantly higher prevalence of Cystic echinococcosis (CE) (33.3%) with 5.3 times (OR 95% 3.5-8.1) than Cysticercus bovis (8.6%) was observed. Significant difference (p < 0.05) in prevalence of both CE and C. bovis were observed among the animal body condition, but not in other factors of the study. From total examined animals, 32.8%, 8.1% and 0.5% were infected by CE only, C. bovis only and both as concurrent infections, respectively. Significantly higher CE (p < 0.05) than C. bovis were observed within studied categories of risk factors. Tissue level cyst prevalence of 83.1% CE and 26.6% C. bovis were observed. High CE counts on the lungs (42.7%), liver (32.8%) and kidney (5.2%) but high C. bovis count in triceps muscle and masseter muscles (7.3% each) and heart (3.64%) were observed. Concurrent infections were observed in heart and liver. Out of the 347 counted CE cysts, fertile and viable were 34.0% and 23.1%, respectively. Of the counted C. bovis, 30.4% were both fertile and viable. A total of 301,139.6ETB (11,155.52USD) annual financial losses were calculated during the study period. The Borana Pastoral Farming System characterized by seasonal animal movement needs awareness creation on the elders and leaders regarding parasitic epidemiology and control strategies.

Spatiotemporal distribution and economic loss associated with bovine cysticercosis and human taeniasis in Ethiopia.

A metacestode stage (bovine cysticercosis) and adult stage Taenia saginata, accounted for zoonotic and economic losses from organ contamination and treatment cost. The objective of this paper is to assess the spatiotemporal distribution and economic loss from bovine cysticercosis and human taeniasis with treatment approach in different parts of Ethiopia. All available published research articles from Ethiopia on the disease were collected. The data were assessed using the current regional administrative, the locality and chronology of the reports for prevalence and economic loss with treatment trend. Prevalence ranging from 1.9% at Addis Ababa City to as high as of 26.3% Hawasa-SNNPR using postmortem examination. However, one study confirmed 92.7% of 41 isolates as bovine cysticercosis using polymerase chain reaction (PCR) method. Teaniasis was ranged from 7.8% in Modjo-Oromia to as high as 89.4% in Addis Ababa City. Except for 17.9% at Addis Ababa City, 29.0% at Mekele-Tigray and 19.0% at Halaba Kulito -SNNPR teaniasis in Ethiopia were greater than 30% over 2001-2018 years. Questionnaire survey and retrospective data were the major tools used for teaniasis reporting either pharmaceutical and/or traditional (herbs) medicines were used for teaniasis treatments. Niclosamide, mebendazole and praziquantel were pharmaceutical teaniacidal drugs used in Ethiopia until the years of 2009, uses of albendazole begins from 2011 to date (2018 year). Hagenica abyssinia "Kosso/Heto" flowers is still widely used in Ethiopia in that it contains Kosso-toxin, closely related to folic acid, are responsible for some cases of hepatic carcinoma and blindness. Only few data (reports) were available on spatiotemporal economic loss associated with bovine cysticercosis and Taenia saginata in Ethiopia showing an overall 4,052,278.16 ETB (212,202.76 USD) from five reports. Highest economic loss (88,500.00 USD) was registered at Yirgalem-SNNPR [2005-2009]. Study at Kombolcha-Amhara region during 2016 accounted 1,841,311.00ETB (73,652.44 USD) from C. bovis and teaniasis treatment cost. Total cost for the used pharmaceutical drugs accounted 921,112.00ETB over 2005-2016. The maximum drug expenditure was reported during the year 2013. Thus, bovine cysticercosis and Taenia saginata in Ethiopia indicating the health and economic challenges. It needs to Taenia saginata cycle interruption via stopping human open defecation with assessing the dynamics of the disease in the country.

Sero-prevalence and community awareness on the risks associated with Livestock and Human brucellosis in selected districts of Fafan Zone of Ethiopian-Somali National Regional State.

A cross-sectional study was conducted to estimate the sero-prevalence, potential risk factors for transmission and spread of brucellosis in livestock and human in Jigjiga and Gursum Woredas of Fafan Zone in Ethiopian-Somali. Two Kebeles were purposively selected from each Worada based on accessibility and willingness of livestock owners. For serology, a total of 268 cattle, 108 sheep, 172 goats, 183 camels, 211 humans were included. For questionnaire, 99 volunteers were recruited. Blood samples were collected from livestock and human. The serum was subjected to Rose Bengal Plate Test (RBPT) and Complement Fixation Test (CFT) to detect Brucella antibody. Out of the total 731 livestock examined, 3.0% were positive for Brucella antibodies using RBPT. Highest sero-prevalence was recorded in camels (4.9%) followed by goat (2.9%), cattle (2.6%), and sheep (0.9%). Using CFT, 0.4% of animals were found positive for brucellosis. A sero-prevalence of 1.7% was recorded in goats using CFT but no in other animal species. From the 211 human serum samples, 5 (2.4%) were positive for Brucella infection using RBPT. One (0.4%) was confirmed by CFT. Questioner survey revealed, almost all respondents (98%) were not aware about zoonotic risks of brucellosis. Cattle and camel milking were mainly performed by housewives. Although 97-99% of respondent had habits of cooked meat consumption, the majorities (99%) consume raw milk. In the pastoral community, the observed sero-prevalence of human brucellosis along with the practices of animal husbandry and animal food consumption habits, might give an insight that brucellosis could pose a public health hazard.

Antimicrobial resistance and genotypic profiles of Salmonella Saintpaul isolated along beef processing and distribution continuum.

Salmonella Saintpaul (SSa) is increasingly reported from food and foodborne outbreak cases. Pulsed field gel electrophoresis (PFGE) is used for screening and tracking of Salmonella infections. Widespread use of antimicrobial agents in humans and food animals could result in antimicrobial resistant Salmonella serotypes. The aim of this study was to characterize S. Saintpaul (n = 28) isolated from various sampling locations at abattoir and meat processing plant lines in Ethiopia for phenotypic antimicrobial resistance and genotypic diversity, and to track its transfer routes. Sampling location, steps and occasions were considered for each isolate description. Antimicrobial sensitivity testing was performed against seven different antimicrobial agents using disc diffusion method. PFGE with XbaI ® enzymatic genomic digestion with BioNumerics® analysis was used for genotypic diversity. Of all the isolates tested, only 17.9% were pan susceptible, and 82.1% were resistant to at least one and at most to three antimicrobials. All isolates were susceptible to gentamycin, trimethoprim-sulfamethoxazol and trimethoprim. Resistance to oxytetracycline (82.2%) was predominant followed by 3.6% resistance to each of chloramphenicol, neomycin and polymyxin B. PFGE analysis revealed three distinguishable clusters of pulsotypes but the majority of the isolates (25/28) belonged to cluster-I (SSaX1-4) pulsotype. Indistinguishable/similar cluster of (SSaX 1-4) isolates among and between sampling location, steps and occasions were observed. Majorities of S. Saintpaul (88%) in the cluster-I pulsotype were resistant to oxytetracycline. Our study indicated that oxytetracycline resistance is very common among the S. Saintpaul isolates studied; and the isolates were diverse with similar resistance profiles within the same genomic pulsotypes. Transfer of S. Saintpaul within, between and across sampling locations, during the same or different occasion were determined from SSaX 1-4 pulsotype while cluster-II (SSaX5) indicates transfer from abattoir to butchery. The unique isolate in cluster-III (SSaX6) shows the presence of other possible source of S. Saintpaul for the beef chain contamination.

Contamination of Ethiopian paper currency notes from various food handlers with E. coli.

Contamination rate of Ethiopian paper currency notes handled by various food handlers with Escherichia coli and antimicrobial susceptibility of the isolates was assessed. A total of 384 Ethiopian Birr (ETB) notes were randomly sampled from meat handlers at butchers, bread and the related food handlers at cafeteria, fruit and vegetables handlers at supermarket, and milk sellers both at open market and dairy station. Fifty control new currencies were also sampled from Commercial Bank of Ethiopia. Both surfaces of the currency were swabbed using wet sterile cotton. The swab was overnight incubated in buffered peptone water. A loop full was streaked on eosin methylene blue agar and followed by biochemical test on presumptive E. coli colonies. Randomly selected isolates were exposed to chloramphenicol (C-30 µg), neomycin (N-30 µg), oxytetracycline (OT-30 µg), polymyxin-B (PB-300 IU) and trimethoprim-sulfamethoxazole (SXT-1.25/23.75/µg) susceptibility using disc diffusion techniques. E. coli was not isolated from currency used as control. A total of 288 (75 %) currency notes were found carrying E. coli. E. coli prevalence was ranges from 67.2 % at open market milk sellers to 87.2 % at dairy station milk sellers; from 64.8 % on ETB 100 to 82.9 % on ETB 1. Differences were not observed in E. coli prevalence on currency notes from among almost all food handlers (P > 0.05). Susceptibility of tested isolates to each chloramphenicol, oxytetracycline and trimethoprim-sulfamethoxazole was 100 %, and to polymyxin-B was 97.3 %. High resistance (83.7 %) was observed to neomycin. The finding indicates, contaminated food can be a source of E. coli for further contamination of currency which again transfer through various foods ready for consumption.

Salmonella serovars along two beef chains in Ethiopia.

INTRODUCTION: Salmonella has been reported from foods and the food production environment, with outbreaks occurring in the human population worldwide. METHODOLOGY: A survey on Salmonella in two beef production lines (a beef abattoir line and a processing line) in Addis Ababa, Ethiopia was conducted, with a total of 668 various samples randomly collected from animal-related materials, the environment, and a beef product (mortadella). RESULTS: Overall, a 12.9% prevalence (26.3% from the abattoir line, 5.3% from the processing plant line) was observed. The prevalence in the abattoir line environment (36.6%) was higher than that in animal-related samples (14.7%); the reverse was true for the processing plant line. Out of 86 isolates, 10 serovars were identified, and 8 remained unidentified. The predominant serotypes were S. Saintpaul (32.5%), S. Muenchen (19.8%), and S. Larochelle (12.8%). S. Kastrup and S. London were isolated for the first time in Ethiopia. CONCLUSIONS: Data indicate open ports of entry for Salmonella, with possible transfer along the line. Further investigations from farm to fork are recommended in order to identify these positions of entry.

Microbiological load and zoonotic agents in beef mortadella from Addis Ababa city supermarkets.

Processing changes the meat ecosystem and, hence, has an impact on the microbiological load and quality of the product. Microbial contamination decreases shelf life and, if a pathogen is present, increases the risk of zoonoses in humans. The aim of this study was to assess the microbiological load, including zoonotic agents, in ready-to-eat beef mortadella from Ethiopian supermarkets. A total of 119 samples from 8 supermarkets in Addis Ababa City (Ethiopia) were examined for aerobic plate count, Enterobacteriaceae, Salmonella, and Escherichia coli. An overall mean of 4.0 log for aerobic plate count and 2.2 log for Enterobacteriaceae was observed. The Salmonella prevalence was 0.8%. An overall E. coli prevalence of 29% (range, 13 to 53%) was observed at individual supermarkets.

Occurrence of Escherichia coli O157:H7 in retail raw meat products in Ethiopia.

BACKGROUND: Escherichia coli O157:H7 (E. coli O157:H7) is now recognized as a major cause of diarrhoea, hemorrhagic colitis and hemolytic-uremic syndrome worldwide. Consumption of raw or undercooked meat of bovine origin has been the most common means of transmitting this organism. METHODOLOGY: Over a period of 7 months (October 2006 to April 2007), a total of 738 raw meat samples were collected from bovines (n= 250), sheep (n=243) and goat (n=245) and investigated for the presence of E. coli O157: H7. Antimicrobial susceptibility testing was performed using the disk diffusion method. RESULTS: E. coli O157:H7 were isolated from 31 (4.2%) out of 738 meat samples examined. Among meat samples examined, the highest prevalence (8%) was recorded in beef, followed by lamb and mutton (2.5%) and goat meat (2%). The isolated strains were found to be susceptible (100%) to amikacin, chloramphenicol, gentamicin, kanamycin, nalidixic acid, norfloxacin, polymyxin B and trimethoprim-sulfamethoxazole. Multidrug resistance to three or more drugs was detected in 7/31 (22.6%) strains. CONCLUSIONS: The results of this study revealed the presence of E. coli O157:H7 in retail raw meats reaching consumers, indicating possible risks of infection to people through the consumption of raw/under-cooked meat or cross-contamination of other food products. Coordinated actions are needed to reduce or eliminate the risks posed by this organism at various stages in food chain. Multiple drug resistant isolates detected in the present study may pose a threat to humans and further limit therapeutic options.