Pathogenic Bacteria and Their Antibiotic Resistance Patterns in Milk, Yoghurt and Milk Contact Surfaces in Debre Berhan Town, Ethiopia.

Background: Bacterial contamination of milk is a primary culprit for causing foodborne illnesses, presenting a significant health hazard for millions of individuals around the globe. The level and variety of microorganisms present in raw milk determine its degree of contamination and the potential health risks it poses. Methods: A cross-sectional survey was conducted from February to August. A questionnaire was used to collect data on socio-demographic characteristics and hygiene practices from milk distributors and traders. Raw milk, yoghurt, swabs from milk containers and drinking cups were collected and processed for bacterial isolation and identification, antibiotic susceptibility testing, MDR screening and confirmation, ESBL screening and confirmation. Finally, all data were pooled and analyzed using SPSS software version 25. Results: A total of 120 samples of fresh milk, yogurt and cotton swabs from milk containers and cups were collected. A total of 80 bacterial isolates were isolated from 120 samples. Among the bacteria isolated, S. aureus 17 (21.3%), E. coli 17 (21.3%), S. epidermidis 14 (17.5%), Klebsiella spp. 9 (11.3%) and Salmonella spp. 7 (8.8%) were detected most often. High rate of contamination was observed in fresh milk 23 (28.8%) and yogurt 23 (28.8%). All isolates were resistant to at least one antibiotic tested. Comparatively, high rates of resistance were observed in all isolates to the most commonly prescribed antibiotics in Ethiopia. However, lower rates of resistance have been observed for recently introduced antibiotics in Ethiopia. Of the isolates, 20 (25.0%) were resistant to eight or more antibiotics. While 16 (20.0%), 12 (15.0%), 9 (11.3%) isolates were resistant to two, three and five antibiotics, respectively. Of the bacteria isolated, 52/80 (65.0%) were MDR, 25/49 (51.0%) were screened for ESBL production, and 20/49 (40.8%) isolates were confirmed as ESBL producer. Conclusion: This study showed a high rate of bacterial isolates along with MDR and ESBL-producing strains in raw milk, yoghurt, milk container swabs and drinking cup swab samples, associated with poor hygiene and sanitation practices.

Prevalence, Antibiotic Susceptibility Pattern, and Associated Factors of Enteric Bacterial Pathogens Among HIV Infected Patients with Diarrhea Attending the ART Clinic of Dilla University Referral Hospital, Southern Ethiopia.

Background: In people with human immunodeficiency virus infection, diarrhea is reportedly associated with significant morbidity and mortality. Therefore, the aim of this study was to determine the prevalence, antibiotic susceptibility pattern, and associated factors of enteric bacterial pathogens among HIV infected patients with diarrhea attending the antiretroviral treatment (ART) clinic of Dilla University Referral Hospital, southern Ethiopia. Methods: This institutional-based cross-sectional study was conducted on 422 study participants attending at ART clinic of Dilla University Referral Hospital from March to August 2022. Demographic and clinical data were collected by using a semi-structured questionnaire. Stool specimens were inoculated on selective media like Butzller's medium and Xylose Lysine Deoxycholate (XLD) agar. Antimicrobial resistance pattern was assessed by using Kirby-Bauer disk diffusion techniques. Adjusted odds ratio (AOR) and 95% Confidence Interval (CI) was used to determine the presence of association. Results: A total of 422 adult patients were enrolled in this study, 51.7% were females. The mean age of the study participants was 27.4 (±15.6 SD) years. The overall prevalence of enteric pathogens was 14.7% (95% CI=11.4-18.2). Shigella spp was the most prevalent organism. Being a farmer (AOR=5.1; 95% CI=1.4-19.1; p<0.015), the habit of hand washing after toilet (AOR=1.9; 95% CI=1.02-3.47; p<0.04), low CD4 cell count of <200 cells (AOR=2.22; 95% CI=1.15-4.27; p<0.02), and longer duration of diarrhea (AOR=2.68; 95% CI=1.23-5.85; p<0.01) were statistically associated. In total, 98.4% of enteric bacterial isolates were sensitive for Meropenem, whereas 82.5% were resistant against Ampicillin. Multidrug resistance was detected in 49.2% of enteric bacteria. Conclusion: We found that enteric bacteria are common causative agents of diarrhea in immune-compromised patients. The high rate of drug resistance calls for escalating antimicrobial susceptibility testing before prescribing antimicrobial agent.

Commonly Consumed Vegetables as a Potential Source of Multidrug-Resistant and ß-Lactamase-Producing Bacteria in Debre Berhan Town, Ethiopia.

Background: Recently, antibiotic resistance of bacteria contained in foods such as vegetables has become a public health problem. In Ethiopia, the diversity of bacterial contamination and level of antibiotic resistance in vegetables are poorly understood. Local analysis of vegetable contamination and its contribution to the spread of antibiotic resistance are therefore essential for One Health interventions. Therefore, the aim of this study was to investigate the level of bacterial contamination of commonly consumed vegetables and their antimicrobial resistance patterns. Methods: A cross-sectional research was conducted in Debre Berhan town from February to August 2022. Questionnaires were used to collect data on sociodemographic variables, hygiene practices, and market hygiene. Six carefully selected vegetables (30 each, 180 in total) were purchased at a local market. Bacterial isolation and identification, multidrug-resistant (MDR) screening and confirmation, extended-spectrum ß-lactamase (ESBL) screening and confirmation, and antibiotic susceptibility tests were performed using standard operating procedure. The data were analysed statistically using SPSS software version 25. Results: The contamination rate of vegetables was 119 (66.1%). Of the 176 bacteria isolates, E. coli (26.1%; 46/176), S. aureus (18.8%; 176), S. epidermidis (10.8%; 19/176), Klebsiella spp. (9.1%; 16/179) and Acinetobacter spp. (6.8%; 12/176) were the most frequently detected isolates. Of the 180 samples tested, (66.1%; 119/180) were contaminated with at least one type of bacteria. Lettuce (22.7%; 40/176), spinach (18.6%; 33/176), and cabbage (19.2%; 32/176) were the most contaminated vegetables. Of the 176 bacteria isolates, (64.8%; 114/176) were MDR, and (18.5%; 23/124) isolates were ESBL producers. The kind of vegetables, vendor/seller finger-nail status, medium of display, market type, and not cleaned before to display were all significantly associated with bacterial contamination. Conclusion: This study found that commonly consumed vegetables are contaminated with antibiotic-resistant bacteria. Vegetables were also notable for the incidence of multidrug-resistant, extended ß-lactamase-resistant, and methicillin-resistant bacterial isolates. Therefore, we urge local health authorities to develop and implement effective control strategies to reduce vegetable contamination.

High Levels of Multidrug-Resistant and Beta-Lactamase-Producing Bacteria in Meat and Meat Contact Surfaces, Debre Berhan Town, Ethiopia.

Background: Over the years, microbial contamination caused by foodborne bacteria has led to a significant number of food recalls, particularly for meat items that have been related to outbreaks of deadly diseases. Animals often carry Salmonella and Escherichia coli bacteria in their intestines, and these bacteria contaminate raw meat during slaughter. In addition, pathogens such as Staphylococcus aureus can contaminate meat processing equipment and utensils and spread to raw meat. Methods: A cross-sectional study was undertaken between 30 February and 15 March 2022. Sanitary conditions of abattoir and butchers and food handlers' hygienic practices were assessed using a structured questionnaire. An equal number of meat, swabs from carcasses, knife, weighing balance and cutting board samples (24 each, 120 total) were collected from abattoir and butcher shops. The collected samples were processed for bacterial isolation, antimicrobial susceptibility testing, MDR screening and confirmation, and ESBL screening and confirmation. Finally, SPSS software version 25 was used to compile and analyze the data. Descriptive data from surveys and laboratory procedures were cross-tabulated and summarized using statistical tables and figure. Results: A total of 76 bacteria were isolated from 120 samples. Of all bacteria isolated, S. aureus 16 (21.1%). E. coli 13 (17.1%), and S. epidermidis 12 (15.8%) were the most prevalent. The rate of bacterial contamination was high in meat 18 (23.7%), carcasses 15 (19.7%) and weighing balance 15 (19.7%), respectively. Among the isolates, 18 (23.7%) were resistant for eight and more antibiotics. While, 17 (22.4%), 7(9.2%) and 4(5.3%) of the isolates were resistant for two and three, four and five, and six and seven antibiotics, respectively. Of bacteria isolated, 51/76 (67.1%) were MDR, 23/48 (47.9%) were screened for ESBL production and 13/48 (27.1%) isolates were confirmed as ESBL producer. Conclusion: Multidrug-resistant bacterial contamination was common in meat and meat contact surfaces, which was exacerbated by inadequate sanitation and hygiene practices.

Parasitic Contamination and Microbiological Quality of Commonly Consumed Fresh Vegetables Marketed in Debre Berhan Town, Ethiopia.

Parasitic and microbial contamination and the pattern of occurrence of the parasite species depends on weather conditions, socio-cultural conditions, sampling season, analyzed vegetable products, and other factors. Therefore, local assessment of vegetable contamination is crucial for targeted and effective interventions. A cross-sectional study was conducted from February to August 2022. A questionnaire was used to assess factors associated with parasite contamination of vegetables during the marketing period. The selected vegetables were purchased and processed for parasite and microbial analysis using standard methods. Finally, all data were summarized and analyzed using SPSS software version 25. A total of 180 vegetable samples were purchased from 180 vendors. This study identified a total of 129 parasites from 180 vegetable samples, with an overall contamination rate of (75; 41.7%). Both protozoa (41; 31.8%) and helminthes (88; 68.2%) were identified from vegetables. Contamination with more than one parasite species was (38; 21.1%). The kind of produce, finger nail status of vendors/sellers, the medium of the display, the type of market and not washed prior to display were significantly associated with parasite contamination. The results also showed that vegetable microbial load for total heterotrophic count, total coliform count, fecal coliform count, yeast count, and mold count was higher in the afternoon than in the morning. To decrease risks to public health, local health authorities and/or market inspectors should establish and implement strategies to reduce contamination such as encouraging specific display medium and washing of vegetables prior to display.

Foodborne Pathogens and Antimicrobial Resistance in Ethiopia: An Urgent Call for Action on "One Health".

Foodborne pathogens are the most common cause of foodborne disease worldwide. They contaminate food at all stages of the food chain, at the agricultural production level (soil and irrigation), from animal sources or at the time of preparation by food handlers. Therefore, food security is a critical issue that affects everyone in the world. Current studies indicate that the problem is more severe in low-income countries like Ethiopia. The majority of studies in Ethiopia have been conducted partially on human, animal or environmental samples with conventional culture-based diagnostic methods. Therefore, this review was conducted to gather information on the main foodborne pathogens and identify gaps in their traceability. Clinically important foodborne pathogens in Ethiopia include (A. lubmbricoids, Toxocara spp., H. nana. E. histolytica/dispar, G. intestinalis, H. diminuta and C. belli), bacteria (Salmonella spp, E. coli O157:H7, B. anthracis, Yersinia, C. perfringens, Klebsiella spp. M. paratuberculosis, and L. monocytogenes), and viruses like (rotaviruses, enteroviruses and astroviruses, hepatitis E virus). In Ethiopia, all isolated foodborne bacterial pathogens showed high rates of antimicrobial resistance (AMR). In particular, the most studied foodborne pathogens, Staphylococcus spp., Salmonella spp., and E. coli from specific sources, showing high levels of resistance to most of the antibiotics prescribed in Ethiopia. The occurrence and persistence of AMR in food is one of the main factors causing the spread of antimicrobial resistance in different compartments, humans, animals and the environment. Therefore, strategies of coordination and struggle from a One Health perspective is an urgent strategy to control antibiotic resistance in order to achieve better outcomes for human and animal health.

High Prevalence of Multi-Drug Resistance and Extended-Spectrum Beta-Lactamase-Producing Enterobacteriaceae Among Hospitalized Patients Presumptive for Bacterial Infection at Debre Berhan Comprehensive Specialized Hospital, Ethiopia.

Background: Multi-drug resistant Enterobacteriaceae (MDR-E), primarily extended-spectrum beta-lactamase producers (ESBLs), have emerged as a major public health concern. This study aimed to determine the prevalence of multi-drug resistance and extended-spectrum beta-lactamase-producing Enterobacteriaceae among hospitalized patients presumptive for bacterial infections at Debre Berhan Comprehensive Specialized Hospital, Ethiopia. Methods: A hospital-based cross-sectional study was conducted from January to May 2021. A total of 384 hospitalized patients presumptive for bacterial infections were included in the study. Urine, wound, blood, stool, and sputum samples were collected and cultured on MacConkey agar, Cysteine Lactose Electrolyte Deficient medium, and Blood agar. Identification was done using a panel of biochemical tests. The antimicrobial susceptibility test was done by disc diffusion. Screening of ESBL production was done by using cefotaxime and ceftazidime and confirmed by the combination disk method per clinical laboratory standard institute guidelines. Data analysis was performed by Statistical Package for Social Sciences software version 25, and a P-value ≤0.05 was considered as statistically significant. Results: Out of 384 study participants, a total of 164 Enterobacteriaceae were isolated. The overall multi-drug resistance rate (MDR) was 92.1%. The overall prevalence of ESBL-PE was 104 (63.4%). E. coli 50 (30.5%) and K. pneumoniae 24 (14.6%) were the predominant ESBL producers. The highest ESBL producers E. coli (13.4%) and K. pneumoniae (6.1%) were isolated from urine sample. History of antibiotic use for the last three months (P-value=0.01), admission in neonatal intensive care unit (P-value=0.02), history of hospital stays (P-value=0.01), and chronic disease (P-value=0.04) showed statistically significant association with ESBL-PE infection. Conclusion: The prevalence of MDR-E and ESBL-PE was high. Therefore, strong infection prevention and control measures and careful selection of antibiotics are needed in the study area to block the transmission and infection in the healthcare setting.

High Magnitude of Fecal Carriage of Extended-Spectrum Beta-Lactamase-Producing Enterobacteriaceae at Debre Berhan Comprehensive Specialized Hospital, Ethiopia.

Background: Gastrointestinal colonization rate of extended-spectrum beta-lactamase-producing Enterobacteriaceae (ESBL-PE) is the major risk factor for infection and dissemination of resistance clones in healthcare facilities. This study aimed to investigate the magnitude of the fecal carriage of ESBL-PE and associated factors among hospitalized patients at Debre Berhan Comprehensive Specialized Hospital, North Shoa, Amhara Regional State, Ethiopia. Methods: A hospital-based cross-sectional study was conducted among 383 hospitalized patients from November 2020 to March 2021. Stool sample or rectal swab was aseptically collected and cultured on different culture media for isolation of Enterobacteriaceae. Identification was done by conventional biochemical tests. Screening of extended-spectrum beta-lactamase (ESBL) production was done by using cefotaxime and ceftazidime and confirmed by the combination disk method. Data analysis was performed by Statistical Package for Social Sciences software version 25 and a P-value ≤0.05 was considered as statistically significant. Results: From the total of 383 hospitalized patients, a total of 347 Enterobacteriaceae were isolated. The overall gastrointestinal colonization rate of ESBL-PE was 47.3% (164/347). The predominant ESBL-PE were E. coli 54.9% (90/164) and K. pneumoniae 33.5% (55/164). The overall multi-drug resistance rate (MDR) was 87.8% (305/347). The highest resistance was observed to ampicillin (98.3%), followed by gentamicin (80.7%), and tetracycline (73.3%), respectively. ESBL-PE were highly susceptible to meropenem (90.2%) and imipenem (89.0%). History of antibiotic use in the past 3 months (p<0.001), admission in the neonatal intensive care unit (p=0.023), and presence of chronic disease (p<0.001) were independently associated with fecal carriage of ESBL-PE. Conclusion: The magnitude of ESBL-PE and MDR was high in the study area. Meropenem and imipenem were active against ESBL-PE. Therefore, strict infection control measure is needed in the study area to limit the infection and dissemination of ESBL-PE.

High Burden of Antibiotic-Resistant Bacteria from Wastewater in Ethiopia: A Systematic Review.

BACKGROUND: Currently, antibiotic-resistant bacteria (ARB) have become a serious global problem and considered as One Health challenge. Despite, wastewater contains a wide range of microbial pathogens and plays a significant role in the dissemination of ARB in the environment. However, it is the most overlooked in developing countries, particularly in Ethiopia. METHODS: Different article searching devices like PubMed, Web of Science, Scopus, and Google Scholar were used to select research article by using the key terms. Hand search using a reference list is also used to retrieve the article. Preferred reporting items for systematic review and meta-analysis (PRISMA) guideline was used for literature search strategy, selection of publications, data extractions, and reporting result for the review process. RESULTS: A total of seven original research articles were included from a total of 35,999 research articles obtained from the different searching techniques. The selected articles were used, the same study design and laboratory methods to isolate different types of resistant bacteria. All studies isolate pathogenic bacteria and highlighted the presence of resistant bacteria for multiple antibiotics. CONCLUSION: Multidrug resistance (MDR) bacteria were isolated from wastewater. This is an indication for the possible presence of pathogenic organisms that are discharged into the receiving environment probably waterbodies (lake and revers) posing risk to public health, animal, and environment. In Ethiopia the coverage of safe water supply is poor.  This obligated the society to use untreated water from rivers, lakes and others. The outlet of most urban sewage from hospital, hotels, and industries are directly running to those water bodies due to lack of proper wastewater management system.  Therefore ARB is a direct threat to those people that use those water bodies.