Isolation and molecular characterization of multi-drug resistant bacteria from cafeterias at the university campus Peshawar

Isolation and molecular characterization of multi-drug resistant bacteria from cafeterias at the university campus Peshawar | Research Square window.SnipcartSettings = { analytics: { enabled: false } }; (function() { var accessVector = localStorage.getItem('access_vector') || ''; window.dataLayer = window.dataLayer || []; if (accessVector) { window.dataLayer.push({ user: { profile: { profileInfo: { snid: accessVector } } } }); } })(); (function(w,d,s,l,i){w[l]=w[l]||[];w[l].push({'gtm.start':new Date().getTime(),event:'gtm.js'});var f=d.getElementsByTagName(s)[0],j=d.createElement(s),dl=l!='dataLayer'?'&l='+l:'';j.async=true;j.src='https://www.googletagmanager.com/gtm.js?id='+i+dl;f.parentNode.insertBefore(j,f);})(window,document,'script','dataLayer','GTM-K279D39R'); Browse Preprints In Review Journals COVID-19 Preprints AJE Video Bytes Research Tools Research Promotion AJE Professional Editing AJE Rubriq About Preprint Platform In Review Editorial Policies Our Team Advisory Board Help Center Sign In Submit a Preprint Cite Share Download PDF Research Article Isolation and molecular characterization of multi-drug resistant bacteria from cafeterias at the university campus Peshawar Jawad Ahmad, Abdullah Jalal This is a preprint; it has not been peer reviewed by a journal. https://doi.org/ 10.21203/rs.3.rs-6700358/v1 This work is licensed under a CC BY 4.0 License Status: Posted Version 1 posted You are reading this latest preprint version Abstract Microbes are the cause of foodborne illnesses, a serious global public health concern that present continuous medical and financial difficulties. In seven student cafeterias at the University Campus Peshawar, the prevalence and patterns of antibiotic resistance of multi-drug resistant bacteria isolated from food contact surfaces were examined. 42 swab samples were taken from utensils, such as plates, forks, spoons, drinking glasses, cutting boards, and washing water. The isolates were identified by molecular characterization by PCR amplification of the 16S rRNA gene and biochemical assays after the samples were cultivated on MacConkey Agar. With an occurrence rate of 44.44%, E. coli had the highest prevalence, followed by S. Typhi (25.93%), S. aureus (18.52%), P. aeruginosae (11.11%), and Enterobacter spp. (7.04%). The highest rates of bacterial contamination were found on cutting boards (37.3%) and in washing water (25.92%), while the lowest rates were found on spoons and plates (7.40%). It was discovered that six multidrug resistant isolates were extremely resistant to chloramphenicol and B-lactam antibiotics. Two multi-drug resistant isolates were identified as E. coli and one as Enterobacter spp. by 16S rRNA sequencing. Ampicillin, Amoxicillin, and Penicillin, which are Β-lactam antibiotics, showed noticeably narrower zones (≤ 9.66 mm). Nalidixic acid and chloramphenicol similarly showed little activity, with high variability in certain isolates (SD = 2.35 and 3.09, respectively). Ciprofloxacin, on the other hand, demonstrated high efficacy by producing the largest inhibitory zones (14 to 21.66 mm) with very low standard deviations (e.g., SD = 0.47–0.81). Different isolates had different zone sizes for Meropenem and Nalidixic Acid, with higher standard deviations (SD = 2.45–3.29 and 2.05–3.09, respectively). Ciprofloxacin (77.8%) was the most effective antibiotic, however the antimicrobial resistance profile of E. coli isolates revealed strong resistance to Penicillin (90.89%), Cefotaxime, and Ceftriaxone (82.44%), suggesting possible ESBL development. S. aureus exhibited strong resistance to Ampicillin (79.22%) and Penicillin (80%), but high susceptibility to Amoxicillin (82.1%), Nalidixic Acid (92%), Meropenem (92.33%), and Ciprofloxacin (92%). S. Typhi showed significant resistance to Cefotaxime and Penicillin (85% each), but considerable susceptibility to Ciprofloxacin (91%) and Nalidixic Acid (84.23%). Meropenem (91%) and Ciprofloxacin (90%) were the most effective antibiotics, however Enterobacter species showed very high resistance to the majority of them. P. aeruginosae exhibited intermediate to high resistance, with the most vulnerable strains being Ciprofloxacin (92.66%) and Nalidixic Acid (70.66%). Introduction Foodborne diseases remain a major global health challenge, with significant morbidity and mortality associated with contaminated food handling environments. Cross-contamination from food contact surfaces has been identified as a critical vector for microbial transmission, particularly of antibiotic-resistant bacteria. In Malaysia and Pakistan, recent studies have shown that utensils such as cutting boards and knives harbor pathogenic and MDR bacteria such as E. coli , Salmonella spp. , and S. aureus . The persistence and adhesion of these bacteria to abiotic surfaces like stainless steel, plastic, and glass make eradication challenging. Moreover, the misuse of antibiotics in clinical and agricultural settings contributes to the emergence of antimicrobial resistance. This study was designed to identify and characterize MDR bacteria from commonly used food contact surfaces and to analyze their resistance profiles to guide improved sanitation and infection control practices. Materials and Methods Sample Collection Samples were collected from six university cafeterias. Swabbing was conducted on cutting boards, utensils (spoons, forks, plates), knives (used and clean), and dishwashing water. A sterile swab pre-moistened with buffered peptone water (BPW) was used for each 25 cm² surface. Samples were transported to the laboratory under chilled conditions. Bacterial Isolation and Identification Swab samples were vortexed, serially diluted, and cultured on selective media: EMB agar for E. coli , Mannitol Salt Agar for S. aureus , XLD for Salmonella spp. , and SS agar for Shigella spp. Colonies were selected for Gram staining, biochemical characterization, and 16S rRNA gene sequencing for molecular confirmation. Antibiotic Susceptibility Testing The Kirby-Bauer disk diffusion method was employed using 12 antibiotics: amikacin, amoxicillin, amoxicillin-clavulanic acid, levofloxacin, minocycline, neomycin, nitrofurantoin, Oxacillin, penicillin G, streptomycin, sulfamethoxazole-trimethoprim, and tetracycline. Results Prevalence of Bacteria Out of all surfaces tested, cutting boards showed the highest contamination levels, especially with E. coli and Salmonella spp . Utensils showed the least contamination. E. coli was detected in 85.36% of samples, predominantly from cutting boards and knives. Salmonella spp. was found in 14.63% of samples. S. aureus and Shigella spp. were also commonly isolated. Antimicrobial Resistance Patterns A large proportion of isolates exhibited resistance to multiple antibiotics: E. coli showed 100% resistance to nitrofurantoin, Oxacillin, penicillin G, and SXT. Salmonella spp. was 100% resistant to tetracycline, penicillin G, and nitrofurantoin. High resistance was also noted to amoxicillin and streptomycin among most isolates. Molecular Identification 16S rRNA sequencing confirmed the identity of E. coli , S. Typhi , S. aureus , and Enterobacter spp . Phylogenetic analysis showed clustering of isolates with known MDR reference strains. Discussion This study reinforces the findings from previous research that food contact surfaces, especially cutting boards and knives, serve as reservoirs for MDR bacteria. The high levels of resistance observed, particularly to first-line antibiotics like penicillin and tetracycline, raise concerns about treatment efficacy and infection control. Moreover, the findings suggest a strong need for enhanced disinfection protocols, staff training in hygiene practices, and reduced antibiotic use in agriculture to mitigate the emergence and spread of MDR bacteria. Conclusion Food contact surfaces in university cafeterias are significantly contaminated with multi-drug resistant bacteria. Cutting boards and knives are the most vulnerable surfaces, highlighting the importance of strict hygiene and regular microbiological monitoring. These findings emphasize the need for targeted interventions to ensure food safety and public health protection. Declarations Author Contribution Abdullah Jalal: Conceived the idea, prepared research protocol and supervised the research work, Provided the technical support and revised the manuscript technically.Jawad Ahmad: Conducted experiments and recorded data, Wrote the first draft, Analyzed the data and prepared results. Acknowledgement Authors are thankful to the Institute of Biotechnology and Genetic Engineering (IBGE), University of Agriculture Peshawar for providing the research facilities. References Zulfakar SS, et al. Microbiological Assessment of Food Contact Surfaces in Residential College Cafeterias. J Sains Kesihatan Malaysia . 2018;16(2):33-38. Ullah R, et al. Antimicrobial resistance in E. coli and Salmonella spp. isolated from table eggs. Microbes and Infectious Diseases . 2023;4(3):968–975. Tsuji M, Yokoigawa K. Attachment of E. coli O157:H7 to Abiotic Surfaces of Cooking Utensils. J Food Sci . 2012;77(4):M194–M197. Eldeeb, H., Serry, F., Ghoname, N. F., & Abbas, H. A. (2020). In vitro activity of tigecycline against local clinical isolates of some Enterobacteriaceae. Zagazig Journal of Pharmaceutical Sciences/Zagazig Journal of Pharmaceutical Science, 29(1), 9. https://doi.org/10.21608/zjps.2020.25134.1007 Grudlewska‐Buda, K., Bauza-Kaszewska, J., Wiktorczyk-Kapischke, N., Budzyńska, A., Gospodarek-Komkowska, E., & Skowron, K. (2023). Antibiotic Resistance in Selected Emerging Bacterial Foodborne Pathogens—An Issue of Concern? [Review of Antibiotic Resistance in Selected Emerging Bacterial Foodborne Pathogens—An Issue of Concern?]. Antibiotics, 12(5), 880. Multidisciplinary Digital Publishing Institute. https://doi.org/10.3390/antibiotics12050880 Gundappa, M., Prabhurajeshwar, C., Ahmed, S., Navya, H. M., Vijayasarathy, M., & Velayuthaprabhu, S. (2021). Microbiological Evaluation and Antibiotic Susceptibility Pattern of Bacterial Isolates Associated with Some Contaminated Edible Fruits and Vegetables. Indian Journal of Agricultural Research. https://doi.org/10.18805/ijare.a-5681 Lamps, L. W. (2003). Pathology of Food-Borne Infectious Diseases of the Gastrointestinal Tract: An Update [Review of Pathology of Food-Borne Infectious Diseases of the Gastrointestinal Tract: An Update]. Advances in Anatomic Pathology, 10(6), 319. Lippincott Williams & Wilkins. https://doi.org/10.1097/00125480-200311000-00002 Miranda, R., & Schaffner, D. W. (2016). Longer Contact Times Increase Cross-Contamination of Enterobacter aerogenes from Surfaces to Food. Applied and Environmental Microbiology, 82(21), 6490. https://doi.org/10.1128/aem.01838-16 Podolak, R., Enache, E., Stone, W. E., Black, D. G., & Elliott, P. H. (2010). Sources and Risk Factors for Contamination, Survival, Persistence, and Heat Resistance of Salmonella in Low-Moisture Foods [Review of Sources and Risk Factors for Contamination, Survival, Persistence, and Heat Resistance of Salmonella in Low-Moisture Foods]. Journal of Food Protection, 73(10), 1919. Elsevier BV. https://doi.org/10.4315/0362-028x-73.10.1919 Urban‐Chmiel, R., Marek, A., Stępień–Pyśniak, D., Wieczorek, K., Dec, M., Nowaczek, A., & Osek, J. (2022). Antibiotic Resistance in Bacteria—A Review [Review of Antibiotic Resistance in Bacteria—A Review]. Antibiotics, 11(8), 1079. Multidisciplinary Digital Publishing Institute. https://doi.org/10.3390/antibiotics11081079 Additional Declarations No competing interests reported. Cite Share Download PDF Status: Posted Version 1 posted You are reading this latest preprint version Research Square lets you share your work early, gain feedback from the community, and start making changes to your manuscript prior to peer review in a journal. As a division of Research Square Company, we’re committed to making research communication faster, fairer, and more useful. We do this by developing innovative software and high quality services for the global research community. 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Also discoverable on Platform About Our Team In Review Editorial Policies Advisory Board Help Center Resources Author Services Accessibility API Access RSS feed Manage Cookie Preferences © Research Square 2026 | ISSN 2693-5015 (online) Privacy Policy Terms of Service Do Not Sell My Personal Information {"props":{"pageProps":{"initialData":{"identity":"rs-6700358","acceptedTermsAndConditions":true,"allowDirectSubmit":true,"archivedVersions":[],"articleType":"Research Article","associatedPublications":[],"authors":[{"id":458834525,"identity":"984ae2df-5d31-4bba-bc6b-af2cbf7dbd8c","order_by":0,"name":"Jawad Ahmad","email":"data:image/png;base64,iVBORw0KGgoAAAANSUhEUgAAAZAAAAAyAQMAAABI0h/eAAAABlBMVEX///8AAABVwtN+AAAACXBIWXMAAA7EAAAOxAGVKw4bAAABDklEQVRIiWNgGAWjYBACxgYgwcNwAMKT/GMDEms8QIKWhjSwGF4tYADXwthwGEzj1cLcfsbww5s/d+TMZzc/fGC547zd2vbDQFtqbKJxOqwnx1hybtszY5k7x4wNJM/cTt52JhGo5VhabgNOv+RukOZtOJw4QyLBTEKC7Xay2QGgFqALcWvpf7v5N8+fw/UzJNK/AbWcSzY7/5CAlhm526R52A4nSEjkmElIth2wM7tByJYZ779Zzm07bDhD5kyxgcSZ5ASzG0BbEvD4xbA/LfnGmz+H5SWk2zc+lqiwszc7n/7wwYcaG9xa4BISwBAH4kSwQAIO5SAgD2cBlTN+YGCwx6N4FIyCUTAKRigAAFquaamhQ5J4AAAAAElFTkSuQmCC","orcid":"","institution":"The University of Agriculture, Peshawar","correspondingAuthor":true,"prefix":"","firstName":"Jawad","middleName":"","lastName":"Ahmad","suffix":""},{"id":458834526,"identity":"70833670-2331-461f-979b-da7d76aa9543","order_by":1,"name":"Abdullah Jalal","email":"","orcid":"","institution":"The University of Agriculture, Peshawar","correspondingAuthor":false,"prefix":"","firstName":"Abdullah","middleName":"","lastName":"Jalal","suffix":""}],"badges":[],"createdAt":"2025-05-19 15:23:32","currentVersionCode":1,"declarations":"","doi":"10.21203/rs.3.rs-6700358/v1","doiUrl":"https://doi.org/10.21203/rs.3.rs-6700358/v1","draftVersion":[],"editorialEvents":[],"editorialNote":"","failedWorkflow":false,"files":[{"id":88624128,"identity":"53a4d931-7e1a-434e-8d3a-cf2f798bf1a2","added_by":"auto","created_at":"2025-08-08 12:32:24","extension":"pdf","order_by":0,"title":"","display":"","copyAsset":false,"role":"manuscript-pdf","size":342296,"visible":true,"origin":"","legend":"","description":"","filename":"manuscript.pdf","url":"https://assets-eu.researchsquare.com/files/rs-6700358/v1/0fed0297-ade0-4654-b82e-46d9668075e1.pdf"}],"financialInterests":"No competing interests reported.","formattedTitle":"Isolation and molecular characterization of multi-drug resistant bacteria from cafeterias at the university campus Peshawar","fulltext":[{"header":"Introduction","content":"\u003cp\u003eFoodborne diseases remain a major global health challenge, with significant morbidity and mortality associated with contaminated food handling environments. Cross-contamination from food contact surfaces has been identified as a critical vector for microbial transmission, particularly of antibiotic-resistant bacteria. In Malaysia and Pakistan, recent studies have shown that utensils such as cutting boards and knives harbor pathogenic and MDR bacteria such as \u003cem\u003eE. coli\u003c/em\u003e, \u003cem\u003eSalmonella spp.\u003c/em\u003e, and \u003cem\u003eS. aureus\u003c/em\u003e.\u003c/p\u003e \u003cp\u003eThe persistence and adhesion of these bacteria to abiotic surfaces like stainless steel, plastic, and glass make eradication challenging. Moreover, the misuse of antibiotics in clinical and agricultural settings contributes to the emergence of antimicrobial resistance. This study was designed to identify and characterize MDR bacteria from commonly used food contact surfaces and to analyze their resistance profiles to guide improved sanitation and infection control practices.\u003c/p\u003e"},{"header":"Materials and Methods","content":"\u003cdiv id=\"Sec3\" class=\"Section2\"\u003e \u003ch2\u003eSample Collection\u003c/h2\u003e \u003cp\u003eSamples were collected from six university cafeterias. Swabbing was conducted on cutting boards, utensils (spoons, forks, plates), knives (used and clean), and dishwashing water. A sterile swab pre-moistened with buffered peptone water (BPW) was used for each 25 cm\u0026sup2; surface. Samples were transported to the laboratory under chilled conditions.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eBacterial Isolation and Identification\u003c/h3\u003e\n\u003cp\u003eSwab samples were vortexed, serially diluted, and cultured on selective media: EMB agar for \u003cem\u003eE. coli\u003c/em\u003e, Mannitol Salt Agar for \u003cem\u003eS. aureus\u003c/em\u003e, XLD for \u003cem\u003eSalmonella spp.\u003c/em\u003e, and SS agar for \u003cem\u003eShigella spp.\u003c/em\u003e Colonies were selected for Gram staining, biochemical characterization, and 16S rRNA gene sequencing for molecular confirmation.\u003c/p\u003e\n\u003ch3\u003eAntibiotic Susceptibility Testing\u003c/h3\u003e\n\u003cp\u003eThe Kirby-Bauer disk diffusion method was employed using 12 antibiotics: amikacin, amoxicillin, amoxicillin-clavulanic acid, levofloxacin, minocycline, neomycin, nitrofurantoin, Oxacillin, penicillin G, streptomycin, sulfamethoxazole-trimethoprim, and tetracycline.\u003c/p\u003e"},{"header":"Results","content":"\u003cdiv id=\"Sec7\" class=\"Section2\"\u003e \u003ch2\u003ePrevalence of Bacteria\u003c/h2\u003e \u003cp\u003eOut of all surfaces tested, cutting boards showed the highest contamination levels, especially with \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eSalmonella spp\u003c/em\u003e. Utensils showed the least contamination.\u003c/p\u003e \u003cp\u003e \u003cem\u003eE. coli\u003c/em\u003e was detected in 85.36% of samples, predominantly from cutting boards and knives.\u003c/p\u003e \u003cp\u003e \u003cem\u003eSalmonella spp.\u003c/em\u003e was found in 14.63% of samples.\u003c/p\u003e \u003cp\u003e \u003cem\u003eS. aureus\u003c/em\u003e and \u003cem\u003eShigella spp.\u003c/em\u003e were also commonly isolated.\u003c/p\u003e \u003c/div\u003e \u003cdiv id=\"Sec8\" class=\"Section2\"\u003e \u003ch2\u003eAntimicrobial Resistance Patterns\u003c/h2\u003e \u003cp\u003eA large proportion of isolates exhibited resistance to multiple antibiotics:\u003c/p\u003e \u003cp\u003e \u003cem\u003eE. coli\u003c/em\u003e showed 100% resistance to nitrofurantoin, Oxacillin, penicillin G, and SXT.\u003c/p\u003e \u003cp\u003e \u003cem\u003eSalmonella spp.\u003c/em\u003e was 100% resistant to tetracycline, penicillin G, and nitrofurantoin.\u003c/p\u003e \u003cp\u003eHigh resistance was also noted to amoxicillin and streptomycin among most isolates.\u003c/p\u003e \u003c/div\u003e\n\u003ch3\u003eMolecular Identification\u003c/h3\u003e\n\u003cp\u003e16S rRNA sequencing confirmed the identity of \u003cem\u003eE. coli\u003c/em\u003e, \u003cem\u003eS. Typhi\u003c/em\u003e, \u003cem\u003eS. aureus\u003c/em\u003e, and \u003cem\u003eEnterobacter spp\u003c/em\u003e. Phylogenetic analysis showed clustering of isolates with known MDR reference strains.\u003c/p\u003e"},{"header":"Discussion","content":"\u003cp\u003eThis study reinforces the findings from previous research that food contact surfaces, especially cutting boards and knives, serve as reservoirs for MDR bacteria. The high levels of resistance observed, particularly to first-line antibiotics like penicillin and tetracycline, raise concerns about treatment efficacy and infection control.\u003c/p\u003e \u003cp\u003eMoreover, the findings suggest a strong need for enhanced disinfection protocols, staff training in hygiene practices, and reduced antibiotic use in agriculture to mitigate the emergence and spread of MDR bacteria.\u003c/p\u003e"},{"header":"Conclusion","content":"\u003cp\u003eFood contact surfaces in university cafeterias are significantly contaminated with multi-drug resistant bacteria. Cutting boards and knives are the most vulnerable surfaces, highlighting the importance of strict hygiene and regular microbiological monitoring. These findings emphasize the need for targeted interventions to ensure food safety and public health protection.\u003c/p\u003e"},{"header":"Declarations","content":"\u003ch2\u003eAuthor Contribution\u003c/h2\u003e\u003cp\u003eAbdullah Jalal: Conceived the idea, prepared research protocol and supervised the research work, Provided the technical support and revised the manuscript technically.Jawad Ahmad: Conducted experiments and recorded data, Wrote the first draft, Analyzed the data and prepared results.\u003c/p\u003e\u003ch2\u003eAcknowledgement\u003c/h2\u003e\u003cp\u003eAuthors are thankful to the Institute of Biotechnology and Genetic Engineering (IBGE), University of Agriculture Peshawar for providing the research facilities.\u003c/p\u003e"},{"header":"References","content":"\u003col\u003e\n\u003cli\u003eZulfakar SS, et al. Microbiological Assessment of Food Contact Surfaces in Residential College Cafeterias. \u003cem\u003eJ Sains Kesihatan Malaysia\u003c/em\u003e. 2018;16(2):33-38.\u003c/li\u003e\n\u003cli\u003eUllah R, et al. Antimicrobial resistance in \u003cem\u003eE. coli\u003c/em\u003e and \u003cem\u003eSalmonella spp.\u003c/em\u003e isolated from table eggs. \u003cem\u003eMicrobes and Infectious Diseases\u003c/em\u003e. 2023;4(3):968\u0026ndash;975.\u003c/li\u003e\n\u003cli\u003eTsuji M, Yokoigawa K. Attachment of \u003cem\u003eE. coli\u003c/em\u003e O157:H7 to Abiotic Surfaces of Cooking Utensils. \u003cem\u003eJ Food Sci\u003c/em\u003e. 2012;77(4):M194\u0026ndash;M197.\u003c/li\u003e\n\u003cli\u003eEldeeb, H., Serry, F., Ghoname, N. F., \u0026amp; Abbas, H. A. (2020). In vitro activity of tigecycline against local clinical isolates of some Enterobacteriaceae. Zagazig Journal of Pharmaceutical Sciences/Zagazig Journal of Pharmaceutical Science, 29(1), 9. https://doi.org/10.21608/zjps.2020.25134.1007\u003c/li\u003e\n\u003cli\u003eGrudlewska‐Buda, K., Bauza-Kaszewska, J., Wiktorczyk-Kapischke, N., Budzyńska, A., Gospodarek-Komkowska, E., \u0026amp; Skowron, K. (2023). Antibiotic Resistance in Selected Emerging Bacterial Foodborne Pathogens\u0026mdash;An Issue of Concern? [Review of Antibiotic Resistance in Selected Emerging Bacterial Foodborne Pathogens\u0026mdash;An Issue of Concern?]. Antibiotics, 12(5), 880. Multidisciplinary Digital Publishing Institute. https://doi.org/10.3390/antibiotics12050880\u003c/li\u003e\n\u003cli\u003eGundappa, M., Prabhurajeshwar, C., Ahmed, S., Navya, H. M., Vijayasarathy, M., \u0026amp; Velayuthaprabhu, S. (2021). Microbiological Evaluation and Antibiotic Susceptibility Pattern of Bacterial Isolates Associated with Some Contaminated Edible Fruits and Vegetables. Indian Journal of Agricultural Research. https://doi.org/10.18805/ijare.a-5681\u003c/li\u003e\n\u003cli\u003eLamps, L. W. (2003). Pathology of Food-Borne Infectious Diseases of the Gastrointestinal Tract: An Update [Review of Pathology of Food-Borne Infectious Diseases of the Gastrointestinal Tract: An Update]. Advances in Anatomic Pathology, 10(6), 319. Lippincott Williams \u0026amp; Wilkins. https://doi.org/10.1097/00125480-200311000-00002\u003c/li\u003e\n\u003cli\u003eMiranda, R., \u0026amp; Schaffner, D. W. (2016). Longer Contact Times Increase Cross-Contamination of Enterobacter aerogenes from Surfaces to Food. Applied and Environmental Microbiology, 82(21), 6490. https://doi.org/10.1128/aem.01838-16\u003c/li\u003e\n\u003cli\u003ePodolak, R., Enache, E., Stone, W. E., Black, D. G., \u0026amp; Elliott, P. H. (2010). Sources and Risk Factors for Contamination, Survival, Persistence, and Heat Resistance of Salmonella in Low-Moisture Foods [Review of Sources and Risk Factors for Contamination, Survival, Persistence, and Heat Resistance of Salmonella in Low-Moisture Foods]. Journal of Food Protection, 73(10), 1919. Elsevier BV. https://doi.org/10.4315/0362-028x-73.10.1919\u003c/li\u003e\n\u003cli\u003eUrban‐Chmiel, R., Marek, A., Stępień\u0026ndash;Pyśniak, D., Wieczorek, K., Dec, M., Nowaczek, A., \u0026amp; Osek, J. (2022). Antibiotic Resistance in Bacteria\u0026mdash;A Review [Review of Antibiotic Resistance in Bacteria\u0026mdash;A Review]. Antibiotics, 11(8), 1079. Multidisciplinary Digital Publishing Institute. https://doi.org/10.3390/antibiotics11081079\u003c/li\u003e\n\u003c/ol\u003e"}],"fulltextSource":"","fullText":"","funders":[],"hasAdminPriorityOnWorkflow":false,"hasManuscriptDocX":true,"hasOptedInToPreprint":true,"hasPassedJournalQc":"","hasAnyPriority":true,"hideJournal":true,"highlight":"","institution":"","isAcceptedByJournal":false,"isAuthorSuppliedPdf":false,"isDeskRejected":"","isHiddenFromSearch":false,"isInQc":false,"isInWorkflow":false,"isPdf":false,"isPdfUpToDate":true,"isWithdrawnOrRetracted":false,"journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true},"keywords":"","lastPublishedDoi":"10.21203/rs.3.rs-6700358/v1","lastPublishedDoiUrl":"https://doi.org/10.21203/rs.3.rs-6700358/v1","license":{"name":"CC BY 4.0","url":"https://creativecommons.org/licenses/by/4.0/"},"manuscriptAbstract":"\u003cp\u003eMicrobes are the cause of foodborne illnesses, a serious global public health concern that present continuous medical and financial difficulties. In seven student cafeterias at the University Campus Peshawar, the prevalence and patterns of antibiotic resistance of multi-drug resistant bacteria isolated from food contact surfaces were examined. 42 swab samples were taken from utensils, such as plates, forks, spoons, drinking glasses, cutting boards, and washing water. The isolates were identified by molecular characterization by PCR amplification of the 16S rRNA gene and biochemical assays after the samples were cultivated on MacConkey Agar. With an occurrence rate of 44.44%, \u003cem\u003eE. coli\u003c/em\u003e had the highest prevalence, followed by \u003cem\u003eS. Typhi\u003c/em\u003e (25.93%), S. aureus (18.52%), P. aeruginosae (11.11%), and Enterobacter spp. (7.04%). The highest rates of bacterial contamination were found on cutting boards (37.3%) and in washing water (25.92%), while the lowest rates were found on spoons and plates (7.40%). It was discovered that six multidrug resistant isolates were extremely resistant to chloramphenicol and B-lactam antibiotics. Two multi-drug resistant isolates were identified as \u003cem\u003eE. coli\u003c/em\u003e and one as Enterobacter spp. by 16S rRNA sequencing. Ampicillin, Amoxicillin, and Penicillin, which are Β-lactam antibiotics, showed noticeably narrower zones (\u0026le;\u0026thinsp;9.66 mm). Nalidixic acid and chloramphenicol similarly showed little activity, with high variability in certain isolates (SD\u0026thinsp;=\u0026thinsp;2.35 and 3.09, respectively). Ciprofloxacin, on the other hand, demonstrated high efficacy by producing the largest inhibitory zones (14 to 21.66 mm) with very low standard deviations (e.g., SD\u0026thinsp;=\u0026thinsp;0.47\u0026ndash;0.81). Different isolates had different zone sizes for Meropenem and Nalidixic Acid, with higher standard deviations (SD\u0026thinsp;=\u0026thinsp;2.45\u0026ndash;3.29 and 2.05\u0026ndash;3.09, respectively). Ciprofloxacin (77.8%) was the most effective antibiotic, however the antimicrobial resistance profile of \u003cem\u003eE. coli\u003c/em\u003e isolates revealed strong resistance to Penicillin (90.89%), Cefotaxime, and Ceftriaxone (82.44%), suggesting possible ESBL development. S. aureus exhibited strong resistance to Ampicillin (79.22%) and Penicillin (80%), but high susceptibility to Amoxicillin (82.1%), Nalidixic Acid (92%), Meropenem (92.33%), and Ciprofloxacin (92%). \u003cem\u003eS. Typhi\u003c/em\u003e showed significant resistance to Cefotaxime and Penicillin (85% each), but considerable susceptibility to Ciprofloxacin (91%) and Nalidixic Acid (84.23%). Meropenem (91%) and Ciprofloxacin (90%) were the most effective antibiotics, however Enterobacter species showed very high resistance to the majority of them. P. aeruginosae exhibited intermediate to high resistance, with the most vulnerable strains being Ciprofloxacin (92.66%) and Nalidixic Acid (70.66%).\u003c/p\u003e","manuscriptTitle":"Isolation and molecular characterization of multi-drug resistant bacteria from cafeterias at the university campus Peshawar","msid":"","msnumber":"","nonDraftVersions":[{"code":1,"date":"2025-05-20 07:37:27","doi":"10.21203/rs.3.rs-6700358/v1","editorialEvents":[{"type":"communityComments","content":0}],"status":"published","journal":{"display":true,"email":"[email protected]","identity":"researchsquare","isNatureJournal":false,"hasQc":true,"allowDirectSubmit":true,"externalIdentity":"","sideBox":"","snPcode":"","submissionUrl":"/submission","title":"Research Square","twitterHandle":"researchsquare","acdcEnabled":true,"dfaEnabled":false,"editorialSystem":"","reportingPortfolio":"","inReviewEnabled":false,"inReviewRevisionsEnabled":true}}],"origin":"","ownerIdentity":"f3c53240-1b70-40c1-b547-409e9f336db3","owner":[],"postedDate":"May 20th, 2025","published":true,"recentEditorialEvents":[],"rejectedJournal":[],"revision":"","amendment":"","status":"posted","subjectAreas":[],"tags":[],"updatedAt":"2025-08-08T12:24:17+00:00","versionOfRecord":[],"versionCreatedAt":"2025-05-20 07:37:27","video":"","vorDoi":"","vorDoiUrl":"","workflowStages":[]},"version":"v1","identity":"rs-6700358","journalConfig":"researchsquare"},"__N_SSP":true},"page":"/article/[identity]/[[...version]]","query":{"redirect":"/article/rs-6700358","identity":"rs-6700358","version":["v1"]},"buildId":"8U1c8b4HqxoKbykW_rLl7","isFallback":false,"isExperimentalCompile":false,"dynamicIds":[84888],"gssp":true,"scriptLoader":[]}