Molecular Features, Epidemiology and Antimicrobial Resistance Profile of Selected Foodborne Bacterial Pathogens Among Diarrheic Patients in Three Selected Cities in Ethiopia
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Date
2025-09-01
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Addis Ababa University
Abstract
Abstract
Background: Foodborne Disease Is a Devastating Global Health Threat, As It May Pose Risks of Serious and Life-Threatening Diseases Along with Chronic Complications. It Disproportionately Affects Low- And Middle-Income Countries Like Ethiopia. However, There Is Fragmented Evidence in Ethiopia on Specific (Mostly Single Pathogen), Restricted Age Groups (Mainly Children Under
Objective: This Study Aimed to Determine the Molecular Features, Epidemiology and Antimicrobial Resistance Profile of Selected Foodborne Bacterial Pathogens (Fbps) Among Diarrheal Patients in Three Selected Cities in Ethiopia. Five Years), Limited Geographic Location, And Season of the Year.
Method: A Cross-Sectional Study Was Conducted from October 2021 to November 2022 in Three Hospitals in Ethiopia (Addis Ababa, Gondar, And Harar). Information On Sociodemographic Characteristics and Clinical Signs and Symptoms Was Collected from Diarrheic Patients Submitting Stool Samples for the Physician Requested Stool Analysis (Microscopic Stool Examination, Culture and Antibiotic Susceptibility Testing) To the Hospitals’ Clinical Laboratories Using a Structured Questionnaire. Stool Samples Were Tested for Shiga-Toxin Producing Escherichia Coli (Stec), Non-Typhoidal Salmonella (Nts), Shigella Spp., And Campylobacter Spp. Using Standardized Conventional Culture and Enzyme Immunoassay Technique. The Stool Samples Were Inoculated on To Selenite F-Broth, And Sub-Cultured on to Xylose Deoxycholate Agar (Xld), And Macconkey Agar (Mac), Followed by the Biochemical Tests (I.E., Triple Sugar Iron Agar, Lysine Iron Agar, Simmon Citrate Agar, Urease Agar, And Sulfide Indole Motility Agar) For the Isolation and Identification of Salmonella and Shigella. The Chromstec Agar Was Used for the Isolation of Stec Characterized by the Mauve Colony Due To Tellurite Resistance. Campylobacter Species Was Detected Using an Enzyme Immunoassay Technique Where Known Enzyme Labeled Antibody Was Used for the Detection of Campylobacter Antigens in the Stool Samples. The Antibiotic Susceptibility Profile of the Isolates Was Determined Using the Phoenix (M50) Method, Using the Broth Microdilution Technique Where Minimum Inhibitory Concentration of Each of the Tested Antibiotics and Their Respective Interpretation Was Provided According to The Inbuilt Clinical and Laboratory Standards Institute (Clsi) and European Committee on Antibiotic Susceptibility Testing (Eucast) Guidelines. Molecular Characterization of Virulence (Stx1, Stx2, Eae, Inva, Stm4200, Sen1392, Ipah, Lacy, 16s Rrna, Hipo, Glya), And the Antibiotic-Resistance (Bla-Tem, Bla-Shv, Bla-Ctx-M, Bla-Oxa-48-Like, Bla-Vim, Bla-Kpc, And Bla-Ndm) Genes Were Conducted Using the Polymerase Chain Reaction. All Statistical Analyses Were Performed Using Rstudio (Version 4.4.3). Univariate Analysis (Descriptive Statistics and Graphics) Was Performed to Summarize the Distribution of Variables. Age Distribution Was Explored Using Box Plots, And Summarized as Means with Standard Deviations, And Median with Interquartile Ranges. Categorical Variables (Socio-Demographic, Clinical, And Environmental Factors) Were Summarized as Frequencies and Proportions. The Prevalence of Targeted Pathogens Was Estimated by Dividing the Total Number of Positive Results for The Targeted Pathogen by the Total Sample Processed Overall, By Study Sites, And by Seasons. Univariable Logistic Regression Was Conducted Between Predictor and the Outcome Variables, For the Crude Odds Ratios With 95% Confidence Intervals Were Calculated. Multivariable Logistic Regression Analyses Were Used to Identify Associated Factors Associated with Occurrence of Pathogens by Controlling the Potential Confounders, And Adjusted Odds Ratio With 95% Confidence Intervals Were Calculated. Variables With P < 0.20 0 In Univariable Analysis Were Included, And Adjusted Odds Ratios (Aor) With 95% Confidence Intervals (Ci) Were Calculated to Quantify Effect Sizes. Multicollinearity Among Explanatory Variables Was Assessed Using the Variance Inflation Factor (Vif) With Values < 10 Considered Acceptable. Variables Exhibiting High Collinearity Would Have Been Considered for Exclusion or Combination to Ensure Model Stability. Results: A Total Of 2,331 Patients Were Enrolled in the Study, With A Median Age Of 28 Years (Range: 2 Months To 94 Years). The Majority of the Study Participants Were Within the Age Group Of 20-29 Years (23.51%) And the Lowest Was in the Age Group Of 15-19 Years (4.59%). All Participants from Addis Ababa Were Urban Residents While About a Quarter of Participants from Harar and Gondar Reported Living in Rural Areas. Most Participants in All Study Sites Were Males (54.78%). The Overall Prevalence of Nts, Stec, Shigella Spp. And Campylobacter Spp. Were 1.29% (95%Ci:0.91,1.84), 12.56% (95%Ci:11.29,13.98), 2.79% (95%Ci:2.19,3.54), And 4.46% (95%Ci:4.61,8.00), Respectively. Moreover, The Prevalence of Eae Harboring Stec Was Found to Be 3.43% (95%Ci:2.77,4.25). Harar Had the Highest Prevalence of All the Pathogens Compared to Addis Ababa and Gondar, For Stec (P<0.001), Nts (P <0.001), Shigella Spp. (P=0.030), And Campylobacter Spp. (P=0.004). Multidrug Resistance Was Detected In 83.33% (25/30) Of Nts, 71.64%Of Shigella Spp., And 48.12% Of Stec Isolates. Moreover, 2.5% Of Stec Isolates Were Extensively Drug-Resistant, All from Harar and Children Under 15 Years. Virulence Profiling of Stec Showed Stx1 In 66.89% Of Isolates, And the Eae Gene In 27.30%. Among Esbl-Positive Stec (N=110), Bla-Ctx-M Predominated (97.12% Overall; 100% In Gondar, 98.31% In Harar, and 86.67% In Addis Ababa), Followed by Bla-Tem (75.00%) And Bla-Shv (15.38%, Highest in Addis Ababa at 26.67%). Moreover, Among the Carbapenem Resistant Stec Isolates, 92.59% And 7.41% Were Found Positive for Bla-Ndm and Bla-Oxa-48-Like Genes Respectively. In Multivariable Logistic Regression Analysis, Foodborne Pathogen Infections Were Significantly Associated with Study Site, Season, Age, Water Source, Sanitation, And Household Animal Ownership. All Predictors Included in the Multivariable Models Had Acceptable Variance Inflation Factor (Vif) Values (<10), Indicating No Problematic Collinearity. Modifiable Factors With The Largest Effect Sizes Included Unimproved Water Sources (Shigella: Aor:1.95, 95%Ci: 1.27,2.74, P = 0.003; Eae-Harboring Stec: Aor:1.06, 95%Ci: 1.01,1.10, P = 0.004), Unimproved Sanitation (Shigella: Aor:1.39, 95%Ci: 1.27,1.74, P = 0.030; Eae-Harboring Stec: Aor:1.08, 95%Ci: 1.01,1.16, P = 0.031), And Household Animal Ownership (Nts: Cattle Aor:1.38, 95%Ci: 1.18,1.46, P = 0.01; Sheep Aor:1.08, 95%Ci: 1.02,1.14, P < 0.001). Non-Modifiable Factors, Including Age, Season, And Study Site, Identified High-Risk Groups and Periods for Targeted Public Health Action
Conclusion: The Findings of This Study Reveal Alarming Levels of Virulent and Drug Resistant Fbps, Particularly in Harar, Posing Urgent Public Health Threats. Targeted Interventions Addressing Seasonal, Environmental, And Antimicrobial Resistance Patterns Are Critical to Control the Spread and Impact of These Infections.
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Molecular Features, Epidemiology and Antimicrobial Resistance Profile, Among Diarrheic Patients