SCREENING AND CHARACTERIZATION OF MULTI DRUG RESISTANT ENTEROCOCCI FROM DOMESTIC EFFLUENTS WITHIN NASARAWA STATE, NIGERIA.
Main Article Content
Abstract
Enterococci are predominantly pathogens inhabiting the intestinal mucosa of animals and humans. The exchange of Plasmid or transposon mobility facilitates horizontal gene transfer among enterococci which favours the development of drug-resistant strains. This study was aimed to determine the frequency and characterize multidrug resistant Enterococci isolated from various domestic effluents in Nasarawa State, Nigeria. A total of 150 samples of domestic effluents from households, dairy farms, restaurants, water factories and bakeries were obtained. Standard cultural and molecular techniques were employed for isolation and identification. Antimicrobial susceptibility profile of Enterococcus species was determined using the Kirby-Bauer disc diffusion method and Multidrug resistance (MDR) was determined. All isolates were cured of their plasmids using acridine orange technique. The percentage occurrence of Enterococcus species across all the five sources were: E. faecalis (78.0%), E. faecium (17.3%), E. durans (1.3%), E. hirae (1.3%) and E. gallinarum (0.6%). MDR for E. faecalis, the highest percentage was found in Household (81.8%) with MDR index of 0.6 while E. faecium recorded the highest percentage in Household (83.3%) with MDR index of 0.7, other species isolated recorded a higher frequency in water factory (100%) with MDR index of 0.9. The presence of plasmid genes (pTEF, vanA, pAD1, pRUM and pAD1) were detected and the isolates’ percentage reduction in antibiotic resistance before and after plasmid curing range from 28.5-100.0 %. The presence of multiple resistant indices in Enterococcus spp. indicates the possibility of outbreaks of pathogenic and multidrug resistant enterococcal infections from domestic effluents.
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Solomon, A., Nfongeh, J. F., Olugbenga, O. O., Akharenegbe , P., Aliyu , A. D., Aliyu , A. D., Eya , B. S., Eya , B. S., & Zakariah, T. (2025). SCREENING AND CHARACTERIZATION OF MULTI DRUG RESISTANT ENTEROCOCCI FROM DOMESTIC EFFLUENTS WITHIN NASARAWA STATE, NIGERIA. The Bioscientist Journal, 13(2), 117-131. https://doi.org/10.54117/the_bioscientist.v13i1.188
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This work is licensed under a Creative Commons Attribution 4.0 International License.
References
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Abimiku R. H., Ngwai Y. B., Nkene I. H., Bassey B. E., Tsaku P. A., Ibrahim T, Tama S. C., Ishaleku D and Pennap G. R. I. (2019). Molecular Diversity and Extended Spectrum Betalactamase Resistance of Diarrheagenic Escherichia coli from Patients Attending Selected Health Care Facilities in Nasarawa State, Nigeria; International Journal of Pathogen Research, 3(1), 1-18.
Adeniji, O. O., Sibanda, T., and Okoh, A. I. (2021). Molecular detection of antibiotic resistance and virulence gene determinants of Enterococcus species isolated from coastal water in the Eastern Cape Province, South Africa. International Journal of Environmental Studies, 78(2), 208-227.
Almeida-Santos, A. C., Novais, C., Peixe, L., and Freitas, A. R. (2021). Enterococcus spp. as a producer and target of bacteriocins: a double-edged sword in the antimicrobial resistance crisis context. Antibiotics, 10(10), 1215-1220.
Amaechi, N. (2015). Plasmid profile and antimicrobial resistance ratings of enterococci isolates from pigs and poultry birds in Abia state, Nigeria. African Journal of Clinical and Experimental Microbiology, 16(2), 54-61.
Armstrong, J. A., and Schulz, J. R. (2015). Agarose gel electrophoresis. Current Protocols Essential Laboratory Techniques, 10(1), 7-2.
Aryal, N., Deng, D., Jha, M. K., and Ofori Boadu, A. (2019). Monitoring, sampling, and automated analysis. Water Environment Research, 91(10), 1288–1293.
Bibalan, M. H., Eshaghi, M., Sadeghi, J., Asadian, M., Narimani, T., and Talebi, M. (2015). Clonal diversity in multi drug resistant (MDR) enterococci isolated from fecal normal flora. International journal of molecular and cellular medicine, 4(4), 240.
Ben Said, L., Hamdaoui, M., Klibi, A., Ben Slama, K., Torres, C., and Klibi, N. (2017). Diversity of species and antibiotic resistance in enterococci isolated from seafood in Tunisia. Annals of microbiology, 67, 135-141.
Boeder, A. M., Spiller, F., Carlstrom, M., and Izídio, G. S. (2024). Enterococcus faecalis: Implications for host health. World Journal of Microbiology and Biotechnology, 40(190), 1-8
Boehm, A. B., and Sassoubre, L. M. (2014). Enterococci as Indicators of Environmental Faecal Contamination. In M. S. Gilmore, D. B. Clewell, Y. Ike, and N. Shankar (Eds.), Enterococci: From Commensals to Leading Causes of Drug-Resistant Infection. Massachusetts Eye and Ear Infirmary.
Boss, R., Overesch, G., and Baumgartner, A. (2016). Antimicrobial resistance of Escherichia coli, Enterococci, Pseudomonas aeruginosa, and Staphylococcus aureus from raw fish and seafood imported into Switzerland. Journal of food protection, 79(7), 1240-1246.
Canton, John, Kate M. Scott, and Paul Glue. (2012): Optimal Treatment of Social Phobia: Systematic Review and Meta-Analysis. Neuropsychiatric Disease and Treatment 8(1) 203-215.
Caneschi, A., Bardhi, A., Barbarossa, A., and Zaghini, A. (2023). The use of antibiotics and antimicrobial resistance in veterinary medicine, a complex phenomenon: A narrative review. Antibiotics, 12(3), 487-493.
Carattoli, A., Lovari, S., Franco, A., Cordaro, G., Di Matteo, P., and Battisti, A. (2005). Extended-Spectrum β-Lactamases in Escherichia coli Isolated from Dogs and Cats in Rome, Italy, from 2001 to 2003. Antimicrobial Agents and Chemotherapy, 49(2), 833–835.
Chahal, C., van den Akker, B., Young, F., Franco, C., Blackbeard, J., and Monis, P. (2016a). Pathogen and Particle Associations in Wastewater. Advances in Applied Microbiology (97), 63–119. Elsevier.
Cho, S., Barrett, J. B., Frye, J. G., and Jackson, C. R. (2020). Antimicrobial Resistance Gene Detection and Plasmid Typing Among Multidrug Resistant Enterococci Isolated from Freshwater Environment. Microorganisms, 8(9), 1338.
Emamalipour, M., Seidi, K., Zununi Vahed, S., Jahanban-Esfahlan, A., Jaymand, M., Majdi, H., Amoozgar, Z., Chitkushev, L. T., Javaheri, T., Jahanban-Esfahlan, R., and Zare, P. (2020). Horizontal Gene Transfer: From Evolutionary Flexibility to Disease Progression. Frontiers in Cell and Developmental Biology, 8, 229-235.
Emurotu, O.M., Ndukwe, K.J., Owemidu, O.I., and Aminu, F.R. (2023). Detection of enterococcus faecalis and antibiotic resistance genes in wastewater facilities from two Nigerian university campuses. Tropical Journal of Natural Product Research, 7(11), 5316–5326.
Farsi, S., Salama, I., Escalante-Alderete, E., and Cervantes, J. (2023). Multidrug-resistant enterococcal infection in surgical patients, what surgeons need to know. Microorganisms, 11(2), 238-243.
Fisher, K., and Phillips, C. (2009). The ecology, epidemiology and virulence of Enterococcus. Microbiology, 155(6), 1749-1757.
Generalov, I. I. (2017). Medical microbiology, virology and immunology. Vitebsk, Belarus: Ministry of Health of the Republic of Belarus Higher Educational Establishment, 587-588.
Gotkowska-Płachta A. (2021). The Prevalence of Virulent and Multidrug-Resistant Enterococci in River Water and in Treated and Untreated Municipal and Hospital Wastewater. International journal of environmental research and public health, 18(2), 563-569.
Green, M. R., and Sambrook, J. (2019). Analysis of DNA by agarose gel electrophoresis. Cold Spring Harbor Protocols, pdb-top100388.
Hashimoto, Y., Suzuki, M., Kobayashi, S., Hirahara, Y., Kurushima, J., Hirakawa, H., and Tomita, H. (2023). Enterococcal linear plasmids adapt to Enterococcus faecium and spread within multidrug-resistant clades. Antimicrobial Agents and Chemotherapy, 67(4), e01619-22.
Humphries, R. M., Ambler, J., Mitchell, S. L., Castanheira, M., Dingle, T., Hindler, J. A., and Sei, K. (2018). CLSI methods development and standardization working group best practices for evaluation of antimicrobial susceptibility tests. Journal of clinical microbiology, 56(4), 1110-1128.
Igwe, J. C., Onaolapo, J. A., Dauda, E. O. and Oladipo, H. O. (2013). Plasmid Conjugation in E. coli and Drug Resistance. Nigerian journal of Biotechnology. 26, 41- 49.
Ikehata, K. (2012). Hazardous agents in wastewater: public health impacts and treatment options for safe disposal and reuse. In Wastewater reuse and management. 165-191. Dordrecht: Springer Netherlands.
Iweriebor, B. C., Gaqavu, S., Obi, L. C., Nwodo, U. U., and Okoh, A. I. (2015). Antibiotic susceptibilities of Enterococcus species isolated from hospital and domestic wastewater effluents in alice, eastern cape province of South Africa. International journal of environmental research and public health, 12(4), 4231–4246.
Kalli, M., Noutsopoulos, C., and Mamais, D. (2023). The fate and occurrence of antibiotic-resistant bacteria and antibiotic resistance genes during advanced wastewater treatment and disinfection: a review. Water, 15(11), 2084-2090.
Khan, N., Ishfaq, M., Mufti, I. U., and Ishfaq, S. (2024). A Short Review on Enterococcus faecalis. Pakistan Journal of Scientific and Industrial Research Series B: Biological Sciences, 67(3).
Kummerer, K. (2003). Significance of antibiotics in the environment. Journal of Antimicrobial Chemotherapy, 52(1), 5–7.
Krause, A. L., Stinear, T. P., and Monk, I. R. (2022). Barriers to genetic manipulation of Enterococci: Current approaches and future directions. Federation of European Microbiology Society. Microbiology Reviews, 46(6), fuac036.
Krawczyk, B., Wityk, P., Gałęcka, M., and Michalik, M. (2021). The many faces of Enterococcus spp. commensal, probiotic and opportunistic pathogen. Microorganisms, 9(9), 1900.
Lebreton, F., Willems, R. J., and Gilmore, M. S. (2014). Enterococcus diversity, origins in nature, and gut colonization. Enterococci: from commensals to leading causes of drug-resistant infection [Internet].
Li, L. G., and Zhang, T. (2023). Plasmid-mediated antibiotic resistance gene transfer under environmental stresses: insights from laboratory-based studies. Science of The Total Environment, 887, 163870.
Ndubuisi, J. C., O. S. Olonitola, A. T. Olayinka, E. D. Jatau, and K. C. Iregbu (2017). Prevalence and Antibiotics Susceptibility Profile of Enterococcus spp. Isolated from Some Hospitals in Abuja, Nigeria. African Journal of Clinical and Experimental Microbiology 18(3): 154.
Partridge, S. R., Kwong, S. M., Firth, N., and Jensen, S. O. (2018). Mobile Genetic Elements Associated with Antimicrobial Resistance. Clinical Microbiology Reviews, 31(4), e00088-17.
Prescott, J. F. (2024). Antimicrobial drug action and interaction: an introduction. Antimicrobial Therapy in Veterinary Medicine, 1-11.
Růžičková, M., Vítězová, M., and Kushkevych, I. (2020). The characterization of Enterococcus genus: Resistance mechanisms and inflammatory bowel disease. Open Medicine, 15(1), 211–224.
Said, M.S., Tirthani, E., and Lesho, E. (2024). Enterococcus Infections. In: StatPearls [Internet]. Treasure Island (FL), StatPearls
Sanderson, H., Ortega-Polo, R., Zaheer, R., Goji, N., Amoako, K. K., Brown, R. S. and McAllister, T. A. (2020). Comparative genomics of multidrug-resistant Enterococcus spp. isolated from wastewater treatment plants. BioMedical Central Journal microbiology, 20, 1-17.
Toru, M., Beyene, G., Kassa, T., Gizachew, Z., Howe, R., and Yeshitila, B. (2018). Prevalence and phenotypic characterization of Enterococcus species isolated from clinical samples of pediatric patients in Jimma University Specialized Hospital, southwest Ethiopia. BioMedical Central Journal research notes, 11, 1-6.
Vanamala, P., Sindhura, P., Sultana, U., Vasavilatha, T., and Gul, M. Z. (2022). Common bacterial pathogens in fish: An overview. Bacterial fish diseases, 279-306.
Waddington, C., Carey, M. E., Boinett, C. J., Higginson, E., Veeraraghavan, B., and Baker, S. (2022). Exploiting genomics to mitigate the public health impact of antimicrobial resistance. Genome medicine, 14(1), 15.
Zaheer, R., Cook, S. R., Barbieri, R., Goji, N., Cameron, A., Petkau, A., and McAllister, T. A. (2020). Surveillance of Enterococcus spp. reveals distinct species and antimicrobial resistance diversity across a One-Health continuum. Scientific reports, 10(1), 3937.
Abimiku, R.H., Ngwai, Y.B., Nkene, I.H., Tatfeng, Y. M. (2016). Molecular detection of diarrheagenic pathotypes of Escherichia coli from diarrheic patients in Keffi, Nigeria. Microbioz Journals, Journal of Microbiology and Biomedical Research, 2(3), 1-6.
Abimiku R. H., Ngwai Y. B., Nkene I. H., Bassey B. E., Tsaku P. A., Ibrahim T, Tama S. C., Ishaleku D and Pennap G. R. I. (2019). Molecular Diversity and Extended Spectrum Betalactamase Resistance of Diarrheagenic Escherichia coli from Patients Attending Selected Health Care Facilities in Nasarawa State, Nigeria; International Journal of Pathogen Research, 3(1), 1-18.
Adeniji, O. O., Sibanda, T., and Okoh, A. I. (2021). Molecular detection of antibiotic resistance and virulence gene determinants of Enterococcus species isolated from coastal water in the Eastern Cape Province, South Africa. International Journal of Environmental Studies, 78(2), 208-227.
Almeida-Santos, A. C., Novais, C., Peixe, L., and Freitas, A. R. (2021). Enterococcus spp. as a producer and target of bacteriocins: a double-edged sword in the antimicrobial resistance crisis context. Antibiotics, 10(10), 1215-1220.
Amaechi, N. (2015). Plasmid profile and antimicrobial resistance ratings of enterococci isolates from pigs and poultry birds in Abia state, Nigeria. African Journal of Clinical and Experimental Microbiology, 16(2), 54-61.
Armstrong, J. A., and Schulz, J. R. (2015). Agarose gel electrophoresis. Current Protocols Essential Laboratory Techniques, 10(1), 7-2.
Aryal, N., Deng, D., Jha, M. K., and Ofori Boadu, A. (2019). Monitoring, sampling, and automated analysis. Water Environment Research, 91(10), 1288–1293.
Bibalan, M. H., Eshaghi, M., Sadeghi, J., Asadian, M., Narimani, T., and Talebi, M. (2015). Clonal diversity in multi drug resistant (MDR) enterococci isolated from fecal normal flora. International journal of molecular and cellular medicine, 4(4), 240.
Ben Said, L., Hamdaoui, M., Klibi, A., Ben Slama, K., Torres, C., and Klibi, N. (2017). Diversity of species and antibiotic resistance in enterococci isolated from seafood in Tunisia. Annals of microbiology, 67, 135-141.
Boeder, A. M., Spiller, F., Carlstrom, M., and Izídio, G. S. (2024). Enterococcus faecalis: Implications for host health. World Journal of Microbiology and Biotechnology, 40(190), 1-8
Boehm, A. B., and Sassoubre, L. M. (2014). Enterococci as Indicators of Environmental Faecal Contamination. In M. S. Gilmore, D. B. Clewell, Y. Ike, and N. Shankar (Eds.), Enterococci: From Commensals to Leading Causes of Drug-Resistant Infection. Massachusetts Eye and Ear Infirmary.
Boss, R., Overesch, G., and Baumgartner, A. (2016). Antimicrobial resistance of Escherichia coli, Enterococci, Pseudomonas aeruginosa, and Staphylococcus aureus from raw fish and seafood imported into Switzerland. Journal of food protection, 79(7), 1240-1246.
Canton, John, Kate M. Scott, and Paul Glue. (2012): Optimal Treatment of Social Phobia: Systematic Review and Meta-Analysis. Neuropsychiatric Disease and Treatment 8(1) 203-215.
Caneschi, A., Bardhi, A., Barbarossa, A., and Zaghini, A. (2023). The use of antibiotics and antimicrobial resistance in veterinary medicine, a complex phenomenon: A narrative review. Antibiotics, 12(3), 487-493.
Carattoli, A., Lovari, S., Franco, A., Cordaro, G., Di Matteo, P., and Battisti, A. (2005). Extended-Spectrum β-Lactamases in Escherichia coli Isolated from Dogs and Cats in Rome, Italy, from 2001 to 2003. Antimicrobial Agents and Chemotherapy, 49(2), 833–835.
Chahal, C., van den Akker, B., Young, F., Franco, C., Blackbeard, J., and Monis, P. (2016a). Pathogen and Particle Associations in Wastewater. Advances in Applied Microbiology (97), 63–119. Elsevier.
Cho, S., Barrett, J. B., Frye, J. G., and Jackson, C. R. (2020). Antimicrobial Resistance Gene Detection and Plasmid Typing Among Multidrug Resistant Enterococci Isolated from Freshwater Environment. Microorganisms, 8(9), 1338.
Emamalipour, M., Seidi, K., Zununi Vahed, S., Jahanban-Esfahlan, A., Jaymand, M., Majdi, H., Amoozgar, Z., Chitkushev, L. T., Javaheri, T., Jahanban-Esfahlan, R., and Zare, P. (2020). Horizontal Gene Transfer: From Evolutionary Flexibility to Disease Progression. Frontiers in Cell and Developmental Biology, 8, 229-235.
Emurotu, O.M., Ndukwe, K.J., Owemidu, O.I., and Aminu, F.R. (2023). Detection of enterococcus faecalis and antibiotic resistance genes in wastewater facilities from two Nigerian university campuses. Tropical Journal of Natural Product Research, 7(11), 5316–5326.
Farsi, S., Salama, I., Escalante-Alderete, E., and Cervantes, J. (2023). Multidrug-resistant enterococcal infection in surgical patients, what surgeons need to know. Microorganisms, 11(2), 238-243.
Fisher, K., and Phillips, C. (2009). The ecology, epidemiology and virulence of Enterococcus. Microbiology, 155(6), 1749-1757.
Generalov, I. I. (2017). Medical microbiology, virology and immunology. Vitebsk, Belarus: Ministry of Health of the Republic of Belarus Higher Educational Establishment, 587-588.
Gotkowska-Płachta A. (2021). The Prevalence of Virulent and Multidrug-Resistant Enterococci in River Water and in Treated and Untreated Municipal and Hospital Wastewater. International journal of environmental research and public health, 18(2), 563-569.
Green, M. R., and Sambrook, J. (2019). Analysis of DNA by agarose gel electrophoresis. Cold Spring Harbor Protocols, pdb-top100388.
Hashimoto, Y., Suzuki, M., Kobayashi, S., Hirahara, Y., Kurushima, J., Hirakawa, H., and Tomita, H. (2023). Enterococcal linear plasmids adapt to Enterococcus faecium and spread within multidrug-resistant clades. Antimicrobial Agents and Chemotherapy, 67(4), e01619-22.
Humphries, R. M., Ambler, J., Mitchell, S. L., Castanheira, M., Dingle, T., Hindler, J. A., and Sei, K. (2018). CLSI methods development and standardization working group best practices for evaluation of antimicrobial susceptibility tests. Journal of clinical microbiology, 56(4), 1110-1128.
Igwe, J. C., Onaolapo, J. A., Dauda, E. O. and Oladipo, H. O. (2013). Plasmid Conjugation in E. coli and Drug Resistance. Nigerian journal of Biotechnology. 26, 41- 49.
Ikehata, K. (2012). Hazardous agents in wastewater: public health impacts and treatment options for safe disposal and reuse. In Wastewater reuse and management. 165-191. Dordrecht: Springer Netherlands.
Iweriebor, B. C., Gaqavu, S., Obi, L. C., Nwodo, U. U., and Okoh, A. I. (2015). Antibiotic susceptibilities of Enterococcus species isolated from hospital and domestic wastewater effluents in alice, eastern cape province of South Africa. International journal of environmental research and public health, 12(4), 4231–4246.
Kalli, M., Noutsopoulos, C., and Mamais, D. (2023). The fate and occurrence of antibiotic-resistant bacteria and antibiotic resistance genes during advanced wastewater treatment and disinfection: a review. Water, 15(11), 2084-2090.
Khan, N., Ishfaq, M., Mufti, I. U., and Ishfaq, S. (2024). A Short Review on Enterococcus faecalis. Pakistan Journal of Scientific and Industrial Research Series B: Biological Sciences, 67(3).
Kummerer, K. (2003). Significance of antibiotics in the environment. Journal of Antimicrobial Chemotherapy, 52(1), 5–7.
Krause, A. L., Stinear, T. P., and Monk, I. R. (2022). Barriers to genetic manipulation of Enterococci: Current approaches and future directions. Federation of European Microbiology Society. Microbiology Reviews, 46(6), fuac036.
Krawczyk, B., Wityk, P., Gałęcka, M., and Michalik, M. (2021). The many faces of Enterococcus spp. commensal, probiotic and opportunistic pathogen. Microorganisms, 9(9), 1900.
Lebreton, F., Willems, R. J., and Gilmore, M. S. (2014). Enterococcus diversity, origins in nature, and gut colonization. Enterococci: from commensals to leading causes of drug-resistant infection [Internet].
Li, L. G., and Zhang, T. (2023). Plasmid-mediated antibiotic resistance gene transfer under environmental stresses: insights from laboratory-based studies. Science of The Total Environment, 887, 163870.
Ndubuisi, J. C., O. S. Olonitola, A. T. Olayinka, E. D. Jatau, and K. C. Iregbu (2017). Prevalence and Antibiotics Susceptibility Profile of Enterococcus spp. Isolated from Some Hospitals in Abuja, Nigeria. African Journal of Clinical and Experimental Microbiology 18(3): 154.
Partridge, S. R., Kwong, S. M., Firth, N., and Jensen, S. O. (2018). Mobile Genetic Elements Associated with Antimicrobial Resistance. Clinical Microbiology Reviews, 31(4), e00088-17.
Prescott, J. F. (2024). Antimicrobial drug action and interaction: an introduction. Antimicrobial Therapy in Veterinary Medicine, 1-11.
Růžičková, M., Vítězová, M., and Kushkevych, I. (2020). The characterization of Enterococcus genus: Resistance mechanisms and inflammatory bowel disease. Open Medicine, 15(1), 211–224.
Said, M.S., Tirthani, E., and Lesho, E. (2024). Enterococcus Infections. In: StatPearls [Internet]. Treasure Island (FL), StatPearls
Sanderson, H., Ortega-Polo, R., Zaheer, R., Goji, N., Amoako, K. K., Brown, R. S. and McAllister, T. A. (2020). Comparative genomics of multidrug-resistant Enterococcus spp. isolated from wastewater treatment plants. BioMedical Central Journal microbiology, 20, 1-17.
Toru, M., Beyene, G., Kassa, T., Gizachew, Z., Howe, R., and Yeshitila, B. (2018). Prevalence and phenotypic characterization of Enterococcus species isolated from clinical samples of pediatric patients in Jimma University Specialized Hospital, southwest Ethiopia. BioMedical Central Journal research notes, 11, 1-6.
Vanamala, P., Sindhura, P., Sultana, U., Vasavilatha, T., and Gul, M. Z. (2022). Common bacterial pathogens in fish: An overview. Bacterial fish diseases, 279-306.
Waddington, C., Carey, M. E., Boinett, C. J., Higginson, E., Veeraraghavan, B., and Baker, S. (2022). Exploiting genomics to mitigate the public health impact of antimicrobial resistance. Genome medicine, 14(1), 15.
Zaheer, R., Cook, S. R., Barbieri, R., Goji, N., Cameron, A., Petkau, A., and McAllister, T. A. (2020). Surveillance of Enterococcus spp. reveals distinct species and antimicrobial resistance diversity across a One-Health continuum. Scientific reports, 10(1), 3937.