Prevalence of Multidrug-Resistant and Biofilm-Forming Salmonella enterica in Beef from Peshawar

Authors

  • Muhammad Jawad Ullah Centre of Biotechnology and Microbiology university of Peshawar, Peshawar, 25000, Khyber Pakhtunkhwa
  • Kafeel Ahmad Centre of Biotechnology and Microbiology University of Peshawar, Peshawar, 25000, Khyber Pakhtunkhwa
  • Faryal Khattak Centre of Biotechnology and Microbiology University of Peshawar, Peshawar, 25000, Khyber Pakhtunkhwa
  • Fawad Inayat Department of Biochemistry, Abdul Wali Khan University, Mardan, Khyber Pakhtunkhwa
  • Sawaira Huriya Centre of Biotechnology and Microbiology University of Peshawar, Peshawar, 25000, Khyber Pakhtunkhwa
  • Naila Menhaj Centre of Biotechnology and Microbiology University of Peshawar, Peshawar, 25000, Khyber Pakhtunkhwa

DOI:

https://doi.org/10.15395/mkb.v58.4138

Keywords:

Antimicrobial resistance, beef contamination, biofilm formation, food safety, Salmonella enterica

Abstract

Salmonella enterica is a major public health concern and is frequently linked tobeef contamination, particularly in developing countries where the emergence of multidrug-resistant (MDR) strains has increased health risks associated with the consumption of contaminated meat. This study aimed to determine the prevalence, seasonal distribution, antimicrobial resistance patterns, and biofilm-forming ability of S. enterica isolated from different points of the beef supply chain in Peshawar, Pakistan. A total of 250 beef samples were collected from retail shops, butcheries, and slaughterhouses across 23 locations between February 2021 and February 2023. The isolates were identified using standard biochemical tests, and antimicrobial susceptibility was evaluated against 16 antibiotics using the Kirby–Bauer disk diffusion method. Biofilm-forming ability was assessed using a quantitative microtiter plate assay. The overall prevalence of S. enterica was 68.0% (170/250), and 150 isolates were further subjected to molecular analysis. A clear seasonal pattern was observed, with the highest prevalence in summer (85.7% in August) and the lowest in winter (43.8% in January), showing a strong positive correlation with temperature (r=0.89, p<0.001). Spatial analysis showed higher odds of contamination near slaughterhouses (OR=2.21, 95% CI: 1.15–4.23) and in urban centers (OR = 2.41, 95% CI: 1.38–4.22) compared to rural areas. High resistance levels were detected against azithromycin (87.3%), tetracycline (61.3%), and streptomycin (48.0%). The MDR phenotype was observed in 56.7% of isolates with a mean MAR index of 0.342. Biofilm formation was detected in 87.3% isolates, and strong producers exhibited a higher prevalence of MDR

References

  1. Han J, Aljahdali N, Zhao S, Tang H, Harbottle H, Hoffmann M, et al. Infection biology of Salmonella enterica. EcoSal Plus. 2024;12(1):eesp00012023. doi:10.1128/ecosalplus.esp-0001–2023
  2. Galán-Relaño Ã, Valero Díaz A, Huerta Lorenzo B, Gómez-Gascón L, Mena Rodríguez MÃ, Carrasco Jiménez E, et al. Salmonella and salmonellosis: an update on public health implications and control strategies. Animals (Basel). 2023;13(23):3666. doi:10.3390/ani13233666
  3. Abebe E, Gugsa G, Ahmed M. Review on major food-borne zoonotic bacterial pathogens. J Trop Med. 2020;2020:4674235. doi:10.1155/2020/4674235
  4. Ehuwa O, Jaiswal AK, Jaiswal S. Salmonella, food safety and food handling practices. Foods. 2021;10(5):907. doi:10.3390/foods10050907
  5. Zeng H, Yang D, Huang N, Li Y, Chen J, Yu Z, et al. Prevalence and antimicrobial susceptibility of salmonella in retail meat collected from different markets in Sichuan, China. Pathogens. 2025;14(3):222. doi:10.3390/pathogens14030222.
  6. Majeed Y, Waseem M. Postharvest Handling Systems. In: Zhang Q, ed. Encyclopedia of smart agriculture technologies. Cham: Springer International Publishing; 2022. p. 1–9. doi: 10.1007/978–3–030–89123–7_125–1
  7. Nazir J, Manzoor T, Saleem A, Gani U, Bhat SS, Khan S, et al. Combatting salmonella: a focus on antimicrobial resistance and the need for effective vaccination. BMC Infect Dis. 2025;25(1):84. doi:10.1186/s12879–025–10478–5
  8. Galán-Relaño Ã, Valero Díaz A, Huerta Lorenzo B, Gómez-Gascón L, Mena Rodríguez MÃ, Carrasco Jiménez E, et al. Salmonella and salmonellosis: an update on public health implications and control strategies. Animals (Basel). 2023;13(23):3666. doi:10.3390/ani13233666
  9. Ahmed MJ, Hossain MI, Bhuiyan MIH, Rahman MA, Kaderi IJ, Nirob AI, et al. Antimicrobial resistance in meat and meat products from Asia: An urgent one health challenge- a systematic review and meta-analysis. Poultry Science. 2025;104(11):105811. doi:10.1016/j.psj.2025.105811
  10. Zhao A, Sun J, Liu Y. Understanding bacterial biofilms: from definition to treatment strategies. Front Cell Infect Microbiol. 2023;13:1137947. doi:10.3389/fcimb.2023.1137947
  11. Sharma S, Mohler J, Mahajan SD, Schwartz SA, Bruggemann L, Aalinkeel R. Microbial biofilm: a review on formation, infection, antibiotic resistance, control measures, and innovative treatment. Microorganisms. 2023;11(6):1614. doi:10.3390/microorganisms11061614.
  12. McDermott PF, Zhao S, Tate H. Antimicrobial resistance in nontyphoidal salmonella. microbiol spectr. 2018;6(4):10.1128/microbiolspec.arba-0014-2017. doi:10.1128/microbiolspec.ARBA-0014–2017
  13. Ferrari RG, Rosario DKA, Cunha-Neto A, Mano SB, Figueiredo EES, Conte-Junior CA. Worldwide epidemiology of salmonella serovars in animal-based foods: a meta-analysis. Appl Environ Microbiol. 2019;85(14). doi:10.1128/AEM.00591-19
  14. Stepanović S, Vuković D, Hola V, Di Bonaventura G, Djukić S, Cirković I, et al. Quantification of biofilm in microtiter plates: overview of testing conditions and practical recommendations for assessment of biofilm production by staphylococci. APMIS. 2007;115(8):891–9. doi:10.1111/j.1600-0463.2007.apm_630.x
  15. Fatima A, Saleem M, Nawaz S, Khalid L, Riaz S, Sajid I. Prevalence and antibiotics resistance status of Salmonella in raw meat consumed in various areas of Lahore, Pakistan. Sci Rep. 2023;13(1):22205. doi: 10.1038/s41598-023-49487-2
  16. Yang X, Huang J, Su Y, Cai S, Zhang J, Guo W, et al. Incidence and antimicrobial resistance of Salmonella serovars in fresh retail aquatic products from China. LWT. 2022;171:114123. doi:10.1016/j.lwt.2022.114123
  17. Lake IR, Gillespie IA, Bentham G, Nichols GL, Lane C, Adak GK, et al. A re-evaluation of the impact of temperature and climate change on foodborne illness. Epidemiol Infect. 2009;137(11):1538–47.
  18. Li B, Webster TJ. Bacteria antibiotic resistance: New challenges and opportunities for implant-associated orthopedic infections. J Orthop Res. 2018;36(1):22–32. doi:10.1002/jor.23656
  19. Manyi-Loh C, Mamphweli S, Meyer E, Okoh A. Antibiotic use in agriculture and its consequential resistance in environmental sources: potential public health implications. Molecules. 2018;23(4):795. doi:10.3390/molecules23040795
  20. Founou LL, Founou RC, Essack SY. Antibiotic resistance in the food chain: a developing country-perspective. Front Microbiol. 2016;7:1881. doi:10.3389/fmicb.2016.01881
  21. Rahman M, Rahman A, Islam M, Alam M. Detection of multi–drug resistant Salmonella from milk and meat in Bangladesh. Bangladesh J Veterinary Med 2018;16(1):115–20. doi:10.3329/bjvm.v16i1.37388
  22. Kumar R, Adeyemi NO, Chattaraj S, Alloun W, Thamarsha A, Anđelković S, et al. Antimicrobial resistance in Salmonella: One Health perspective on global food safety challenges. Sci One Health. 2025;4:100117. doi:10.1016/j.soh.2025.100117.
  23. Xia Y, Hu Z, Jin Q, Chen Q, Zhao C, Qiang R, et al. Structural characteristics, functions, and counteracting strategies of biofilms in Staphylococcus aureus. Comput Struct Biotechnol J. 2025;27:488–500. doi:10.1016/j.csbj.2025.01.021.
  24. Aleksandrowicz A, Carolak E, Dutkiewicz A, BÅ‚achut A, Waszczuk W, Grzymajlo K. Better together-Salmonella biofilm-associated antibiotic resistance. Gut Microbes. 2023;15(1):2229937. doi:10.1080/19490976.2023.222993
  25. Carrascosa C, Raheem D, Ramos F, Saraiva A, Raposo A. Microbial biofilms in the food industry-a comprehensive review. Int J Environ Res Public Health. 2021;18(4):2014. doi:10.3390/ijerph18042014
  26. Samtiya M, Matthews KR, Dhewa T, Puniya AK. Antimicrobial resistance in the food chain: trends, mechanisms, pathways, and possible regulation strategies. Foods. 2022;11(19):2966. doi:10.3390/foods11192966

Downloads

Published

2026-03-31

Issue

Vol. 58 No. 1 (2026)

Section

Articles

License

This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License (CC BY-NC 4.0).

Under this license, authors retain copyright and grant Majalah Kedokteran Bandung the right of first publication. Others may share, remix, and adapt the work for non-commercial purposes, provided that appropriate credit is given to the author and to Majalah Kedokteran Bandung as the initial place of publication.

For details, please see: https://creativecommons.org/licenses/by-nc/4.0/

 

How to Cite

Prevalence of Multidrug-Resistant and Biofilm-Forming Salmonella enterica in Beef from Peshawar. (2026). Majalah Kedokteran Bandung, 58(1), 1-9. https://doi.org/10.15395/mkb.v58.4138