Investigation of Antibiotic-Resistant Enterococci in Farm Animals and Vegetables: A case study of Selected Towns in Ekiti State, Nigeria
Keywords:
Antimicrobial resistance, Enterococci, Animals, Food-vegetables, EkitiAbstract
Enterococci can be found in unprocessed and ready-to-eat farm produce. The extensive use of antimicrobials in agricultural practices has influenced natural selection in Enterococci. Human consumers can be infected with these antibiotic-resistant strains with resulting clinical complications. In this study, the resistance and virulence potential of Enterococcus spp. from farm animals and food-vegetables in Ekiti, Nigeria was investigated. Ninety-six samples: poultry rectal swabs (n = 35), swine faecal droppings (n = 18) and vegetables (n= 43) were processed for the isolation of Enterococcus spp. Isolated Enterococcus spp. were further identified by different biochemical tests. Susceptibility to antibiotics was investigated using the Kirby-Bauer disk diffusion test on Mueller Hinton agar. A total number of 118 isolates were obtained from which 96 (81%) Enterococciwere recovered, with 63 (66%) E. faecium, while 33 (34%) were identified as other species. Twenty-eight (28%) of the isolates tested were susceptible to Augmentin; resistance to cloxacillin was observed among all isolates while resistance to gentamicin, ofloxacin, vancomycin and erythromycin was observed among 97%, 82%, 75%, and 72%, respectively. Thirty-nine (41%) exhibited the ability to produce biofilms. Ninety-three (97%) isolates expressed multiple drug resistance to three or more antimicrobials with 15 different antimicrobial resistance patterns. AUG/CXC/ERY/GEN/OFL/VAN was the most predominant (n = 36) resistant pattern. This study therefore, emphasises farm animals and food-vegetables to be important carriers of Enterococcus spp., with a high prevalence of multidrug resistance, which could be transferred through the food chain implicatively serving as threat to the public health.
References
Aarestrup, F. M, Kruse, H., Tast, E, Hammerum, A. M. and Jensen, L. B. (2000). Associations between the use of antimicrobial agents for growth promotion and the occurrence of resistance among Enterococcus-faecium from broilers and pigs in Denmark, Finland, and Norway. Microbial Drug Resistance; 6: 63-70.
Adebowale, O. O., Bankole, M. and Olasoju, M. (2020) Farmers’ perceptions and drivers of antimicrobial use and abuse in commercial pig production, Ogun State, Nigeria. Int J Environ Res Public Health.7. 10.3390/ijerph-17103579.
Adesokan, H. K., Akanbi, I. O.,Akanbi, I. M. and Obaweda R. A, (2015). Pattern of antimicrobial usage in livestock animals in south-western Nigeria. The need for alternative plans. Onderstepoort Journal of Veterinary Research; 82: 816.
Alemayehu, T., Hailemariam, M. (2020). Prevalence of Vancomycin-resistant enterococcus in Africa in one health approach: a systematic review and meta-analysis. Sci Rep., 10, 20542.
Almansour, A. M., Meshari, A., Alhadlaq, K. O., Alzahrani, L. E., Mukhtar, A. L. and Sulaiman M. A. (2023). The Silent Threat: Antimicrobial-Resistant Pathogens in Food - Producing Animals and Their Impact on Public Health. Microorganisms11, no. 9: 2127.
Amaechi, N. and Nwankwo, I.U. (2015). Evaluation of prevalence and antimicrobial resistance using enterococci isolates from pigs and poultry birds in Abia State, Nigeria, International Journal Current Microbiology Applied Science, 4, 825-833
Anyanwu, M.U. and Obetta, T. U. (2015). Prevalence and antibiogram of generic enterococci in ready-to-slaughter beef cattle. NotulaeScientia Biologicae. 7, 390 – 399.
Ayeni, F.A., Odumosu, B.T., Oluseyi, A.E. and Ruppitsch, W. (2016). Identification and prevalence of tetracycline resistance in enterococci isolated from poultry in IIishan, Ogun State, Nigeria. Journal of Pharmacy and Bioallied Sciences, 8, 69 – 73.
Badul, S., Abia, A. L. K., Amoako, D. G., Perrett, K., Bester, L. A., and Essack, S. Y. (2021). From the Farms to the Dining Table: The Distribution and Molecular Characteristics of Antibiotic-Resistant Enterococcus spp. in Intensive Pig Farming in South Africa. Microorganisms.; 9(5): 882.
Barreto, A., Guimaraes, B., Radhouani, H., Araujo, C., Goncalves, A., Gaspar, E., Rodrigues, J., Igrejas, G. and Poeta, P. (2009). Detection of antibiotic-resistant E. coli and Enterococcus spp. in stool of healthy growing children in Portugal. Journal of Basic Microbiology.49: 503–512.
Bashiru, A., Igbinosa, I. H., Omeje, F. I., Ogofure, A. G., Eyong, M. M., and Igbinosa, E. O. (2017). Multi-antibiotic resistant and putative virulence gene signatures in Enterococcus species isolated from pig farms environment. Microb. Pathog.; 104: 90–96.
Bates, J., Jordens, Z. and Selkon, J. B. (1993). Evidence for an animal origin of Vancomycin-resistant Enterococci. Lancet: 342: 490-491.
Ben Sallem, R., Klibi, N., Klibi, A., Ben Said, L., Dziri, R., Boudabous, A., Torres, C. and Ben Slama K, (2016). Antibiotic resistance and virulence of Enterococci isolates from healthy humans in Tunisia. Annals of Microbiology; 66: 717-725.
Bhardwaj, S. B. (2019). Enterococci: An Important Nosocomial Pathogen. In: Kirmusaoglu S (Ed), Pathogenic Bacteria. Intech Open. Rijeka, Croatia, p. 1-17
Brinkwirth, S., Ayobami, O., Eckmanns, T. and Markwart, R. (2021). Hospital-acquired infections caused by enterococci: a systematic review and meta-analysis, WHO European Region, 1 January 2010 to 4 February 2020. Euro Surveillance. 26(45): 2001628.
Castaño-Arriba, A., González-Machado, C., Igrejas, G., Poeta P., Alonso-Calleja, C., and Capita, R. (2020). Antibiotic Resistance and Biofilm-Forming Ability in Enterococcal Isolates from Red Meat and Poultry Preparations. Pathogens.; 9(12): 1021.
Cebeci, T. (2024). Species prevalence, virulence genes, and antibiotic resistance of enterococci from food-producing animals at a slaughterhouse in Turkey. Scientific Reports.14: 13191.
Chung, Y.S., Kwon, K.H., Shin, S., Kim, J.H., Park, Y.H., and Yoon, J.W., (2014). Characterization of veterinary hospital associated isolates of Enterococcus species in Korea, Journal of Microbiology and Biotechnology. 24: 386 – 393.
Costa, L. F. X., Falcão, D. A., Grassotti, T. T., Christiano, F. D. P., Frazzon, J. and G. Frazzon, A. P. G. (2022). Antimicrobial resistance of enterococci isolated from food in South Brazil: Comparing pre- and post-RDC 20/2011. An Acad Bras Cienc., 94: e20201765.
Dolka, B., Gołębiewska–Kosakowska, M., Krajewski, K., Kwieciński, P., Nowak, T., Szubstarski, J., Wilczyński, J., and Szeleszczuk, P. (2017). Occurrence of Enterococcus spp. in poultry in Poland based on 2014–2015 data. Med. Water.; 73: 220–224.
Eldaly, E. A., Rasha, M., and Elazam, R. A. (2019). Prevalence of Enterococcus species in chicken meat in Sharkia Governorate. Egypt. J. Appl. Sci.; 34: 317–323.
Emeje, M. O., Oloye, S., Bubakari, M., and Adeyeye M. (2022). Use of antibiotics in livestock in Nigeria: Curbing antimicrobial resistance and developing a national regulatory guideline towards monitoring antibiotic use in animal and animal foods. International Research Journal of Public and Environmental Health; Vol.9 (2): pp. 55-66.
Feiyang, M., Shixin, X., Zhaoxin, T., Zekun, L., and Lu, Z. (2021). Use of antimicrobials in food animals and impact of transmission of antimicrobial resistance on humans, Biosafety and Health. 3(1): 32-38.
Grassotti, T. T., Zvoboda, D. A., Costa, L. F. X., Araújo, A. J. G., Pereira, R. I., Soares, R. O., Wagner, P. G. C., Frazzon, J., and Frazzon, A. P. G. (2018). Antimicrobial resistance profiles in Enterococcus spp. isolates from fecal samples of wild and captive black capuchin monkeys (Sapajusnigritus) in South Brazil. Frontiers in Microbiology. 9: 2366.
Grudlewska-Buda, K., Skowron, K., Bauza-Kaszewska, J., Budzyńska, A., Wiktorczyk-Kapischke, N., Wilk, M., Wujak, M., and Paluszak, Z. (2023). Assessment of antibiotic resistance and biofilm formation of Enterococcus species isolated from different pig farm environments in Poland. BMC Microbiology. 23(1):89.
Hammerum, A. M., Lester, C. H., Neimann, J.et al. (2004). A Vancomycin-resistant Enterococcusfaecium isolate from a Danish healthy volunteer, detected 7 years after the ban of avoparcin, is possibly related to pig isolates. Journal of Antimicrobial Chemo-therapy; 53: 547-549.
Hollenbeck, B. L., and Rice, L. B. (2012). Intrinsic and acquired resistance mechanisms in Enterococcus. Virulence, 3: 421–569.
Iweriebor, B. C., Obi, L. C., and Okoh, A. I., (2015). Virulence and antimicrobial resistance factors of Enterococcus spp. isolated from faecal samples from piggery farms in Eastern Cape, South Africa. BMC Microbiology; 15: 1-136.
Iweriebor, B. C., Obi, L. C., and Okoh, A. I., (2016). Macrolide, glycopeptide resistance and virulence genes in Enterococcus species isolates from dairy cattle. Journal of Medical Microbiology; 65: 641-648.
Jeżak, K., Kozajda, A. (2022). Occurrence and spread of antibiotic-resistant bacteria on animal farms and in their vicinity in Poland and Ukraine - review. Environ Science and Pollution Research. 29: 9533–9559.
Kim, H. N., Kim, W. H., Park, M. S., Seo, H. G., Cho, J. T., Yu, R. H., Kim, H. S., Hwang, H. J., Choi, C., and Rhee, S. M. (2020). Virulence patterns and prevalence of seven Enterococcus species isolated from meats and leafy vegetables in South Korea. Food Control, Volume 108: 106867, ISSN 0956-7135.
Klibi, A., Ben, Lagha, A., Ben Slama, K., Boudabous, A., and Torres, C. (2013). Faecal Enterococci from camels in Tunisia: species, antibiotic resistance and virulent genes. Veterinary Records. 172: 213.
Kühn, I., Iversen, A., Finn, M., Greko, C., Burman, L. G., Blanch, A. R., Vilanova, X., Manero, A., Taylor, H., Caplin, J., Domínguez, L., Herrero, I. A., Moreno, M. A., and Möllby, R. (2005). Occurrence and relatedness of Vancomycin-resistant enterococci in animals, humans, and the environment in different European regions. Applied and Environmental Microbiology. 71(9):5383-90.
Kwon, K. H., Hwang, S. Y., Moon, B. Y., Park, Y. K., Shin, S., Hwang, C. Y., and Park, Y. H., (2012). Occurrence of antimicrobial resistance and virulence genes, and distribution of enterococcal clonal complex 17 from animals and human beings in Korea. Journal of Veterinary Diagnostic Investigation; 24: 924-931.
Molechan, C., Amoako, D. G., Abia, A. L. K., Somboro, A. M., Bester, L. A., and Essack, S. Y. (2019). Molecular epidemiology of antibiotic-resistant Enterococcus spp. from the farm-to-fork continuum in intensive poultry production in KwaZulu-Natal, South Africa. Sci. Total Environ.;692: 868–878.
Mudenda, S., Matafwali, S. K., Malama, S., Munyeme, M., Yamba, K., Katemangwe, P., Siluchali, G., Mainda, G., Mukuma, M., Bumbangi, F.N., et al. (2022). Prevalence and antimicrobial resistance patterns of Enterococcus species isolated from laying hens in Lusaka and Copperbelt provinces of Zambia: A call for AMR surveillance in the poultry sector. JAC-Antimicrob. Resist.; 4:126.
Mukuna, W., Aniume, T., Pokharel, B., Khwatenge, C., Basnet, A., &Kilonzo-Nthenge, A. (2023). Antimicrobial Susceptibility Profile of Pathogenic and Commensal Bacteria Recovered from Cattle and Goat Farms. Antibiotics (Basel, Switzerland), 12(2), 420. https://-doi.org/10.3390/antibiotics12020420
Mwikuma, G., Kainga, H., Kallu, S. A., Nakajima, C., Suzuki, Y., and Hang'ombe, B. M. (2023). Determination of the Prevalence and Antimicrobial Resistance of Enterococcusfaecalis and Entero-coccusfaecium Associated with Poultry in Four Districts in Zambia. Antibiotics (Basel).; 12(4): 657.
Ngbede, E. O., Raji, M. A., Kwanashie, C. N., and PajhiKwaga, J. K. (2016). Antimicrobial resistance and virulence profile of Enterococci isolated from poultry and cattle sources in Nigeria. Tropical Animal Health Production; pp5.
Novais, C., Freitas, A. R., Silveira, E., Antunes, P., Silva, R., Coque, T. M., and Peixe, L. (2013). Spread of multidrug-resistant Enterococcus to animals and humans: An underestimated role for the pig farm environment. J. Antimicrob. Chemother., 68: 2746–2754.
O’Driscoll, T. and Crank, C.W., (2015). Vancomycin-resistant enterococcal infections: epidemiology, clinical manifestation and optimal management, Infection and Drug Resistance, 8: 217 – 230.
Oguntoyinbo, F. A., and Okueso, O. (2013). Prevalence, distribution and antibiotic resistance pattern among enterococci species in two traditional fermented dairy foods. Annals of Microbiology, 63: 55.
Olonitola, O. S., Fahrenfeld, N., and Pruden, A., (2015). Antibiotic resistance profile among mesophilic aerobic bacteria in Nigerian chicken litter and associated antibiotic resistance genes. Poultry Science; 94: 867-874.
Oluwasile, B. B., Agbaje, M., Ojo, O. E., and Dipeolu, M. A., (2014). Antibiotic usage pattern in selected poultry farms in Ogun state. Sokoto Journal of Veterinary Science. 12: 45-50.
Orababa, O. Q., Soriwei, J. D., Akinsuyi, S. O., Essiet, U. U., and Solesi, O. M. (2021). A systematic review and meta-analysis on the prevalence of Vancomycin-resistant enterococci (VRE) among Nigerians. Porto Biomedical Journal. 6(1):e125.
Osafo, R., Balali, I. G., Amissah-Reynolds, P., Gyapong, F., Addy, R., Nyarko, A. A., and Wiafe, P. (2022). Microbial and Parasitic Contamination of Vegetables in Developing Countries and Their Food Safety Guidelines. Journal of Food Quality. 2022 (1): 4141914.
Osman, K.M., Ali, M.N., Radwan, I., ElHofy, F., Abed, A.H., Orabi, A., and Fawzy, N.M., (2016). Dispersion of the Vancomycin resistance genes van A and van C of Enterococcus isolated from Nile Tilapia on retail sale: A public health hazard. Frontiers in Microbiology.7: 1354.
Popović, N., Dinić, M., Tolinački, M., Mihajlović, S., Terzić-Vidojević, A., Bojić, S., Djokić, J., Golić, N., and Veljović, K. (2018). New Insight into Biofilm Formation Ability, the Presence of Virulence Genes and Probiotic Potential of Enterococcus sp. Dairy Isolates. Frontiers in Microbiology: 9 -2018.
Prieto, A. M. G., Van Schaik, W., Rogers, M. R. C., Coque, T. M., Baquero, F., Corander, J., and Willems, R. J. L. (2016). Global emergence and dissemination of enterococci as nosocomial pathogens: attack of the clones? Front Microbiol7: 788.
Pruksakom, C., Pimam, C., Boonsoongnem, A., and Narongsak, W., (2016). Detection and phenotypic characterization of Vancomycin-resistant Enterococcus in pigs in Thailand, Agriculture and Natural Resources. 50: 199 – 203.
Rahman, M., Alam, M. U., Luies, S. K., Kamal, A., Ferdous, S., Lin, A., Sharior, F., Khan, R., Rahman, Z., Parvez, S. M., Amin, N., Hasan, R., Tadesse, B. T., Taneja, N., Islam,M. A., and Ercumen, A. (2021). Contamination of Fresh Produce with Antibiotic-Resistant Bacteria and Associated Risks to
Human Health: A Scoping Review. International Journal of EnvironmentalResearch and Public Health. 30; 19(1):360.
Ramos, S., Silva, V., Dapkevicius, M., Igrejas, G., and Poeta, P. (2020). Enterococci, from Harmless Bacteria to a Pathogen. Microorganisms. 8(8):1118.
Ristori, C.A., Rowlands, R.E.G., Bergamini, M.M., Lopes, G.I.S.L., de Paula, M.R., de Olivera, M.A., Lima, M., Tegani, L.S., Watanabe, A.H., Jakabi, M., and Zanella, R.C., (2012). Prevalence and antimicrobial susceptibility profile of Enterococcus species isolated from frozen chicken carcasses. Revista do Instituto Adolfo Lutz (Impresso),.71: 237 – 243.
Steoien-Pysniak, D., Marek, A., Banach, T., Adaszek, L., Pysik, E., Wilczyrisk, J., and Winiarcyk, S., (2016). Prevalence and antibiotic resistance of Enterococcus strains isolated from poultry. ActaVeterinariaHungarica; 64.
Tan, S. C., Chong, C. W., Teh, C. S., Ooi, P. T., and Thong, K. L. (2018). Occurrence of virulent multidrug-resistant Enterococcusfaecalis and Enteroco-ccusfaecium in the pigs, farmers and farm environments in Malaysia. Peer J.; 6: e5353.
U.S. Environmental Protection Agency. (2024). Indicators: Enterococci. U.S. Environmental Protection Agency. Available online at: https://-www.epa.gov/national-aquatic-re-source-surveys/indicators-enterococci
Urakawa, H., Hancock, T., Kratz, M., and Armstrong, R. (2021). QT-AMP: Quanti-Tray-based amplicon sequencing for simultaneous quantification and identification of enterococci for microbial source tracking. bioRxiv. https://doi.org/-10.1101/2021.08.02.454799.
Van Boeckel, T. P., Brower, C., Gilbert, M., Grenfell, B. T., Levin, S. A., Robinson, T. P., et al., (2015). Global trends in antimicrobial use in food animals. Proc Natl Acad Sci U. S. A, 112: pp. 5649-5654.
Wegener, H. C., (1998). Historical yearly usage of glycopeptides for animals and humans: the American–European paradox revisited. Antimicrobial Agents and Chemotherapy. 42: 3049.
WHO (2024). WHO's List of Medically Important Antimicrobials for Human Medicine: a risk management tool for mitigating antimicrobial resistance due to non-human use; World Health Organization: Geneva: World Health Organization; 2024. Licence: CC BY-NC-SA 3.0 IGO.
Wist, V.,Morach, M.,Schneeberger, M.,Cernela, N., Stevens, M.J.A.,Zurfluh, K., Stephan, R.. and Nüesch-Inderbinen, M. (2020). Phenotypic and genotypic traits of Vancomycin-resistant enterococci from healthy food-producing animals. Microorganisms. 8: 261.
Woolhouse, M., Ward, M., van Bunnik, B. and Farrar, J., (2015). Antimicrobial resistance in humans, livestock and the wider environment. Philosophical Transactions of the Royal Society; B: 370.
World Health Organization. (2023). Antimicrobial resistance. World Health Organization. https://www.-who.int/news-room/fact-sheets/-detail/antimicrobial-resistance
Xu, W., Fang, Y., and Zhu, K. (2023). Enterococci facilitate polymicrobial infections. Trends in Microbiology. 32 (2): 162-177.
Yilmaz, E.S., Aslantas, O., Onen, S.P., Turkylmaz, S., and Kurekci, C. (2016). Prevalence, antimicrobial resistance and virulence traits in enterococci from food of animal origin in Turkey, LWT - Food Science and Technology, 66: 20 – 26
Downloads
Published
How to Cite
Issue
Section
License
Copyright (c) 2024 Researchers Journal of Science and Technology
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
