Evaluation of minimum inhibitory concentration and multi-drug resistance profile of Salmonella isolated from layer poultry farms in kaski district, Nepal

Suresh Nepali; Sharada  Thapaliya; Narayan Paudyal; Jeevan  Adhikari; Rajesh Gautam

doi:10.64636/ar.19

Authors

Suresh Nepali Faculty of Animal Science, Veterinary Science and Fisheries, Agriculture and Forestry University, Rampur Chitwan, Rampur Chitwan 44209, Nepal

Competing Interests

.The authors declare no conflict of interest.
Sharada Thapaliya Faculty of Animal Science, Veterinary Science and Fisheries, Agriculture and Forestry University, Rampur Chitwan, Rampur Chitwan 44209, Nepal https://orcid.org/0000-0001-7548-5362

Competing Interests

.
Narayan Paudyal Nepal Agriculture Research Council, National Animal Health Research Centre, Khumaltar, Kathmandu 44700, Nepal https://orcid.org/0000-0001-8204-2674

Competing Interests

.
Jeevan Adhikari Faculty of Animal Science, Veterinary Science and Fisheries, Agriculture and Forestry University, Rampur Chitwan, Rampur Chitwan 44209, Nepal

Competing Interests

.
Rajesh Gautam Institute of Agriculture and Animal Science, Tribhuvan University, Paklihawa, Rupandehi 33007, Nepal https://orcid.org/0009-0006-9733-5238

Competing Interests

.

Corresponding Author

Rajesh Gautam

vet.rajesh01@gmail.com ; https://orcid.org/0009-0006-9733-5238

DOI:

https://doi.org/10.64636/ar.19

Abstract

Salmonella species are the most prevalent bacterial pathogens infecting poultry. Antibiotics are administered as growth promoters or a treatment in poultry, raising the prospect of antibiotic resistance. There are very few baselines quantitative studies on antibiotic Minimum Inhibitory Concentration (MIC) against Salmonella of poultry origin. Thus, this study aims to evaluate the MIC and multi-drug resistance (MDR) profile of Salmonella isolated from the poultry farms of Kaski district of Nepal. Laboratory analysis was conducted to determine the minimum inhibitory concentration of nineteen different antibiotic molecules from various classes against Salmonella spp. isolated from poultry feces. The bacteria were identified using standard protocols and confirmed with polymerase chain reaction for the invA gene. Positively confirmed isolates were subjected to a MIC evaluation assay by the E-Test protocol, using Ezy MIC Strips following EUCAST guidelines and interpreted accordingly. Out of 610 samples collected, on screening using the invA gene for Salmonella, 48 isolates were confirmed as Salmonella spp., with an overall farm prevalence of 34.43% (n=61) and a sample-level prevalence of 7.8%. Salmonella spp. displayed the highest resistance to colistin (100%), ciprofloxacin (88%), norfloxacin (72%), ceftriaxone (72%), amoxicillin (77%), ampicillin (66%), erythromycin (66%), cefotaxime (66%), tetracycline (66%), doxycycline (55%), and chloramphenicol (55%). However, amikacin (94%) showed higher susceptibility. MIC analysis revealed elevated values for colistin, ciprofloxacin, amoxicillin, ampicillin, ceftriaxone, and doxycycline compared to epidemiological cut-offs. Multiple drug resistance was observed in 94% of Salmonella spp. isolates, each showing unique multiple antibiotic resistance (MAR) patterns. The study highlights the concerns of antibiotic resistance in commercial layer farms in Kaski district, possibly due to excessive antibiotic use by poultry farm owners. Urgent measures are needed to address this public health issue by regulating antibiotic usage in the poultry industry in the area.

Keywords:

Antibiotic resistance, Poultry, Salmonella spp., minimum inhibitory concentration, multi-drug resistance

Author Biography

Jeevan Adhikari, Faculty of Animal Science, Veterinary Science and Fisheries, Agriculture and Forestry University, Rampur Chitwan, Rampur Chitwan 44209, Nepal

Department of Veterinary Surgery and Pharmacology, Faculty of Veterinary Science, Agriculture and Forestry University, Rampur Chitwa

References

Acharya, K. P., Subramanya, S. H., & Lopes, B. S. (2019). Combatting antimicrobial resistance in Nepal: The need for precision surveillance programmes and multi-sectoral partnership. In JAC-Antimicrobial Resistance, 1(3). Oxford University Press, UK. https://doi.org/10.1093/jacamr/dlz066

Acharya, K. P., & Wilson, R. T. (2019). Antimicrobial Resistance in Nepal. Frontiers in Medicine, 6(May), 7–9. https://doi.org/10.3389/fmed.2019.00105

Agbaje, M., Begum, R. H., Oyekunle, M. A., Ojo, O. E., & Adenubi, O. T. (2011). Evolution of Salmonella nomenclature: A critical note. Folia Microbiologica, 56(6), 497–503. https://doi.org/10.1007/s12223-011-0075-4

Bantawa, K., Sah, S. N., Subba Limbu, D., Subba, P., & Ghimire, A. (2019). Antibiotic resistance patterns of Staphylococcus aureus, Escherichia coli, Salmonella, Shigella and Vibrio isolated from chicken, pork, buffalo and goat meat in eastern Nepal. BMC Research Notes, 12(1), 1–6. https://doi.org/10.1186/s13104-019-4798-7

Basnyat, B., Pokharel, P., Dixit, S., & Giri, Sjj. (2015). Antibiotic use, its resistance in Nepal and recommendations for action: a situation analysis. https://doi.org/10.33314/jnhrc.v0i0.632.

Cheesbrough, M. (2006). District Laboratory Practice in Tropical Countries (2nd ed.). Cambridge University Press. https://doi.org/10.1017/CBO9780511543470

Citron, D. M., Ostovari, M. I., Karlsson, A., & Goldstein, E. J. C. (1991). Evaluation of the E test for susceptibility testing of anaerobic bacteria. Journal of Clinical Microbiology, 29(10), 2197–2203. https://doi.org/10.1128/jcm.29.10.2197-2203.1991

DLS. (2025). Livestock and fisheries statistics of Nepal 2080/81 (2023/24). Animal Genetic Resources and Economic Analysis Division, Livestock Statistics Management and Economic Analysis Section, Department of Livestock Services, Ministry of Agriculture and Livestock Development. https://dls.gov.np/

Fowler, P. D., Sharma, S., Pant, D. K., Singh, S., & Wilkins, M. J. (2021). Antimicrobial-resistant non-typhoidal Salmonella enterica prevalence among poultry farms and slaughterhouses in Chitwan, Nepal. Veterinary World, 14(2), 437–445. https://doi.org/10.14202/vetworld.2021.437-445

Gautam, N., Poudel, R., Lekhak, B., & Upreti, M. K. (2019). Antimicrobial Susceptibility Pattern of Gram-Negative Bacterial Isolates from Raw Chicken Meat Samples. Tribhuvan University Journal of Microbiology, 6(1), 89–95. https://doi.org/10.3126/tujm.v6i0.26590

Gyawali, A. (2018). Antimicrobial Resistance Profiling of Salmonella enterica Isolated from Commercial Poultry of Chitwan, Nepal Arrogya Gyawali (Research Associate) Poultry Conference 2018.

Hossain, Md. J., Attia, Y., Ballah, F. M., Islam, Md. S., Sobur, Md. A., Islam, Md. A., Ievy, S., Rahman, A., Nishiyama, A., Islam, Md. S., Hassan, J., & Rahman, Md. T. (2021). Zoonotic Significance and Antimicrobial Resistance in Salmonella in Poultry in Bangladesh for the Period of 2011–2021. Zoonotic Diseases, 1(1), 3–24. https://doi.org/10.3390/zoonoticdis1010002

Ibrahim, S., Wei Hoong, L., Lai Siong, Y., Mustapha, Z., C. W. Zalati, C. W. S., Aklilu, E., Mohamad, M., & Kamaruzzaman, N. F. (2021). Prevalence of antimicrobial resistance (AMR) Salmonella spp. and escherichia coli isolated from broilers in the east coast of peninsular malaysia. Antibiotics, 10(5). https://doi.org/10.3390/antibiotics10050579

Iwamoto, M., Reynolds, J., Karp, B. E., Tate, H., Fedorka-Cray, P. J., Plumblee, J. R., Hoekstra, R. M., Whichard, J. M., & Mahon, B. E. (2017). Ceftriaxone-resistant nontyphoidal salmonella from humans, retail meats, and food animals in the United States, 1996-2013. Foodborne Pathogens and Disease, 14(2), 74–83. https://doi.org/10.1089/fpd.2016.2180

Jajere, S. M., Hassan, L., Zakaria, Z., Abu, J., & Aziz, S. A. (2020). Antibiogram profiles and risk factors for multidrug resistance of salmonella enterica recovered from village chickens (Gallus gallus domesticus linnaeus) and other environmental sources in the central and Southern Peninsular Malaysia. Antibiotics, 9(10), 1–13. https://doi.org/10.3390/antibiotics9100701

Koirala, R., Panthee, N., Pradhan, S., Rajbhandari, N., Shrestha, D. K., Chhetri, S., Shrestha, Y., Dahal, A., Dhakal, S., & Thapa, S. (2020). Multi-drug Resistant and Extended Spectrum β-lactamase Producing Salmonella Species Isolated from Fresh Chicken Liver Samples. Kathmandu University Medical Journal (KUMJ), 18(70), 133–138. https://doi.org/10.3126/kumj.v18i1.33342

Kowalska-Krochmal, B., & Dudek-Wicher, R. (2021). The minimum inhibitory concentration of antibiotics: Methods, interpretation, clinical relevance. Pathogens 10(2), 1 –21. https://doi.org/10.3390/pathogens10020165

Kuehn, B. M. (2022). News From the Centers for Disease Control and Prevention. JAMA - Journal of the American Medical Association, 327(14), 1324. https://doi.org/10.1001/jama.2022.4829

Kumar, D., Pornsukarom, S., & Thakur, S. (2019). Antibiotic Usage in Poultry Production and Antimicrobial-Resistant Salmonella in Poultry. In Food Safety in Poultry Meat Production (pp. 47–66). Springer International Publishing. https://doi.org/10.1007/978-3-030-05011-5_3

Maharjan, A., Dhungel, B., Bastola, A., Shrestha, U. T., Adhikari, N., Banjara, M. R., Lekhak, B., Ghimire, P., & Rijal, K. R. (2021a). Antimicrobial susceptibility pattern of salmonella spp. isolated from enteric fever patients in Nepal. Infectious Disease Reports, 13(2), 388–400. https://doi.org/10.3390/idr13020037

Maharjan, A., Dhungel, B., Bastola, A., Shrestha, U. T., Adhikari, N., Banjara, M. R., Lekhak, B., Ghimire, P., & Rijal, K. R. (2021b). Antimicrobial susceptibility pattern of salmonella spp. isolated from enteric fever patients in Nepal. Infectious Disease Reports, 13(2), 388–400. https://doi.org/10.3390/idr13020037

Majumder, M. A. A., Rahman, S., Cohall, D., Bharatha, A., Singh, K., Haque, M., & Gittens-St Hilaire, M. (2020). Antimicrobial stewardship: Fighting antimicrobial resistance and protecting global public health. Infection and Drug Resistance, 13, 4713–4738. https://doi.org/10.2147/IDR.S290835

Mąka, Ł., & Popowska, M. (2016). Antimicrobial resistance of Salmonella spp. isolated from food. Roczniki Panstwowego Zakladu Higieny, 67(4), 343–358.

Nair, D. V. T., Venkitanarayanan, K., & Johny, A. K. (2018). Antibiotic-resistant Salmonella in the food supply and the potential role of antibiotic alternatives for control. Foods, 7(10). https://doi.org/10.3390/foods7100167

Nelson, A., Manandhar, S., Ruzante, J., Gywali, A., Dhakal, B., Dulal, S., Chaulagai, R., & Dixit, S. M. (2020). Antimicrobial drug resistant non-typhoidal Salmonella enterica in commercial poultry value chain in Chitwan, Nepal. One Health Outlook, 2(1). https://doi.org/10.1186/s42522-020-00025-4

Pal, P., Bhattarai, R.K., Bhatta, R., Acharya, P., Singh, S., & Harries, A.D. (2022). Antimicrobial resistance in bacteria isolated from the poultry production system in Nepal. The Public Health Action, Dec21;12(4):165-170 doi:10.5588/pha.22.0014

Prajapati, M., Ranjit, E., Shrestha, R., Shrestha, S. P., Adhikari, S. K., & Khanal, D. R. (2018). Status of antibiotic residues in poultry meat of Nepal. Nepalese Veterinary Journal, 35, 55–62. https://doi.org/https://doi.org/10.3126/nvj.v35i0.25240

Sharma, A., & Tripathi, P. (2015). Seasonal Prevalence of Poultry Diseases and their interaction with IBD and Mycotoxicosis in Chitwan, Nepal. The Research Gate, August 2015. https://doi.org/10.13140/RG.2.2.27054.41283

Sharma, S., Fowler, P. D., Pant, D. K., Singh, S., & Wilkins, M. J. (2021). Prevalence of non-typhoidal Salmonella and risk factors on poultry farms in Chitwan, Nepal. Veterinary World, 14(2), 426–436. https://doi.org/10.14202/vetworld.2021.426-436

Shrestha, A., Regmi, P., Dutta, R. K., Khanal, D. R., Aryal, S. R., Thakur, R. P., Karki, D., & Singh, U. M. (2010). First report of antimicrobial resistance of Salmonella isolated from poultry in Nepal. Veterinary Microbiology, 144(3–4), 522–524. https://doi.org/10.1016/j.vetmic.2010.04.015

Tenover, F. C. (2006). Mechanisms of Antimicrobial Resistance in Bacteria. American Journal of Medicine, 119(6 SUPPL. 1). https://doi.org/10.1016/j.amjmed.2006.03.011

Upadhyaya, N., Karki, S., Oli, M., & Tiwari, R. N. (2020). Status of antimicrobial use in livestock sector in Nepal. Department of Livestock Services, Ministry of Agriculture and Livestock Development, Government of Nepal Ministry: Katmandu, Nepa.

Upadhyaya, N., Karki, S., Rana, S., Elsohaby, I., Tiwari, R., Oli, M., & Paudel, S. (2023a). Trend of Antimicrobial Use in Food-Producing Animals from 2018 to 2020 in Nepal. Animals, 13(8). https://doi.org/10.3390/ani13081377

Upadhyaya, N., Karki, S., Rana, S., Elsohaby, I., Tiwari, R., Oli, M., & Paudel, S. (2023b). Trend of Antimicrobial Use in Food-Producing Animals from 2018 to 2020 in Nepal. 1–11.

Wright, G. D. (2007). The antibiotic resistome: The nexus of chemical and genetic diversity. Nature Reviews Microbiology, 5(3), 175–186. https://doi.org/10.1038/nrmicro1614

Zhai, L., Kong, X., Lu, Z., Lv, F., Zhang, C., & Bie, X. (2014). Detection of Salmonella enterica serovar Dublin by polymerase chain reaction in multiplex format. Journal of Microbiological Methods, 100(1), 52–57. https://doi.org/10.1016/j.mimet.2014.02.014

Ziaul Haque, A. K. M., Akter, M. R., Islam, S. K. S., Alam, J., Neogi, S. B., Yamasaki, S., & Lutful Kabir, S. M. (2021). Salmonella gallinarum in small-scale commercial layer flocks: Occurrence, molecular diversity and antibiogram. Veterinary Sciences, 8(5). https://doi.org/10.3390/vetsci8050071

Type: Article

Evaluation of minimum inhibitory concentration and multi-drug resistance profile of Salmonella isolated from layer poultry farms in kaski district, Nepal

Authors

Corresponding Author

DOI:

Abstract

Keywords:

Author Biography

References

View and Download

Downloads

Published

Competing Interest

Data Availability Statement

Issue

Section

Categories

License

How to Cite

Share

Downloads

Most read articles by the same author(s)

Similar Articles

Journal Highlights

Indexing and Abstracting

Digital Preservation

Animal Reports’ Commitment to Long-Term Digital Preservation

Keywords

Reviewer Credits

Browse