EVALUATION OF VIRULENCE AND ANTIMICROBIAL RESISTANCE GENES OF ENTEROCOCCUS SPECIES ISOLATED FROM SHEEP MILK WITH SUBCLINICAL MASTITIS

Authors

DOI:

https://doi.org/10.71336/jabs.1253

Keywords:

Antimicrobial resistance, Enterococcus faecalis, Enterococcus faecium, sheep mastitis, virulence genes

Abstract

This study aimed to investigate the formation of antimicrobial resistance genes associated with vancomycin and gentamicin, as well as the capacity to develop biofilms, in Enterococcus sp. strains isolated from subclinical mastitis in sheep. In total, Enterococcus faecalis (22/26, 84.62%) was found to be the most prevalent species among the samples of mastitic milk collected from sheep. In general, E. faecalis strains exhibited higher resistance compared to Enterococcus faecium. After isolation and identification using polymerase chain reaction, a total of twenty-six enterococci (22 E. faecalis, 4 E. faecium) were submitted to susceptibility tests against eight antimicrobial agents. The isolates indicated multidrug resistance to amoxicillin-clavulanic acid (100%), kanamycin (100%), and trimethoprim-sulfamethoxazole (100%). All isolates were susceptible to ampicillin (100%), and florfenicol (100%) and vancomycin susceptibility was observed in 53.85% of the isolates. The dominant antimicrobial resistance genes detected in our isolates were vanB (19.23%), aph(2”)-Ia1 (61.54%), aph(2”)-Id1 (26.92%), and esp (65.38%) as the most common virulence gene. This research indicated a significant occurrence of antimicrobial resistance in E. faecalis and E. faecium strains obtained from subclinical sheep mastitis. The increasing resistance of enterococci to antibiotics poses a growing challenge to both human well-being and the environment.

References

Ahmed, W., Neubauer, H., Tomaso, H., El Hofy, F. I., Monecke, S., Abd El-Tawab, A. A., Hotzel, H. (2021): Characterization of Enterococci and ESBL-Producing Escherichia coli Isolated from Milk of Bovides with Mastitis in Egypt. Pathogens (Basel, Switzerland) 10: 97. DOI: https://doi.org/10.3390/pathogens10020097

Krawczyk, B., Wityk, P., Gałęcka, M., Michalik, M. (2021): The Many Faces of Enterococcus spp. - Commensal, Probiotic and Opportunistic Pathogen. Microorganisms 9: 1900. DOI: https://doi.org/10.3390/microorganisms9091900

García-Solache, M., Rice, L. B. (2019): The Enterococcus: A Model of Adaptability to Its Environment. Clinical Microbiology Reviews 32: 10-1128. DOI: https://doi.org/10.1128/CMR.00058-18

Gao, X., Fan, C., Zhang, Z., Li, S., Xu, C., Zhao, Y., Liu, M. (2019): Enterococcal Isolates from Bovine Subclinical and Clinical Mastitis: Antimicrobial Resistance and Integron-Gene Cassette Distribution. Microbial Pathogenesis 129: 82-87. DOI: https://doi.org/10.1016/j.micpath.2019.01.031

Różańska, H., Lewtak-Piłat, A., Kubajka, M., Weiner, M. (2019): Occurrence of Enterococci in Mastitic Cow’s Milk and Their Antimicrobial Resistance. Journal of Veterinary Research 63: 93. DOI: https://doi.org/10.2478/jvetres-2019-0014

El-Zamkan, M. A., Mohamed, H. M. (2021): Antimicrobial Resistance, Virulence Genes and Biofilm Formation in Enterococcus Species Isolated from Milk of Sheep and Goat with Subclinical Mastitis. Plos One 16: e0259584. DOI: https://doi.org/10.1371/journal.pone.0259584

de Freitas, G. F., Felipe, F., Fernandes, J. S., Trevizan, G. S., Sossai, P. F. A., de Figueiredo, P. J. C., Helio, L. (2023): Antimicrobial Susceptibility Profile of Enterococcus Species İsolated from Cows with Clinical Mastitis and from Bulk Milk Tanks in Brazil. African Journal of Microbiology Research 17: 29-34. DOI: https://doi.org/10.5897/AJMR2022.9657

Savci, Ü., Şahin, M., Eser. B. (2018): Klinik Örneklerden İzole Edilen Enterococcus faecalis ve Enterococcus faecium Suşlarında Antibiyotik Dirençlerinin Değerlendirilmesi. Journal of Health Sciences and Medicine 1: 4-8. DOI: https://doi.org/10.32322/jhsm.405716

Azizi, M., Hasanvand, B., Kashef, M., Alvandi, A. H., Abiri, R. (2017): Virulence Factor and Biofilm Formation in Clinical Enterococcal Isolates of the West of Iran. Jundishapur Journal of Microbiology 10: e14379. DOI: https://doi.org/10.5812/jjm.14379

Davis, B. C., Keenum, I., Calarco, J., Liguori, K., Milligan, E., Pruden, A., Harwood, V. J. (2022): Towards the Standardization of Enterococcus Culture Methods for Waterborne Antibiotic Resistance Monitoring: A Critical Review of Trends Across Studies. Water Res X. 19: 100161. DOI: https://doi.org/10.1016/j.wroa.2022.100161

da Silva Fernandes, M., Alvares, A. C. C., Manoel, J. G. M., Esper, L. M. R., Kabuki, D. Y., Kuaye, A.Y. (2017): Formation of Multi-Species Biofilms by Enterococcus faecium, Enterococcus faecalis, and Bacillus cereus Isolated From Ricotta Processing and Effectiveness of Chemical Sanitation Procedures. International Dairy Journal 72: 23-28. DOI: https://doi.org/10.1016/j.idairyj.2017.03.016

Suvarna, K., Mahon, C. R. (2022): Streptococcus, Enterococcus, and Other Catalase-Negative, Gram-Positive Cocci. Textbook of Diagnostic Microbiology-E-Book, 7th Ed., Elsevier Publishing, Amsterdam, Netherland.

Walaa, F. A., Felemban, E. M., Shafie, A., Alhomrani, M., Habeeballah, H., Alsharif, K., Farid, M. A. (2019): The Antimicrobial Resistance and Prevalence of Enterococcus Species in Saudi Arabia. Journal of Pure and Applied Microbiology 13: 2461-2470. DOI: https://doi.org/10.22207/JPAM.13.4.61

El-Mahdy, R., Mostafa, A., El-Kannishy, G. (2018): High Level Aminoglycoside Resistant Enterococci in Hospital-Acquired Urinary Tract Infections in Mansoura, Egypt. Germs, 8: 186. DOI: https://doi.org/10.18683/germs.2018.1145

Moosavian, M., Ghadri, H., Samli, Z. (2018): Molecular Detection of VanA and VanB Genes Among Vancomycin-Resistant Enterococci in ICU-Hospitalized Patients in Ahvaz in Southwest of Iran. Infection and Drug Resistance 11: 2269-2275. DOI: https://doi.org/10.2147/IDR.S177886

Pandey, R., Mishra, S. K., Shrestha, A. (2021): Characterisation of ESKAPE Pathogens with Special Reference to Multidrug Resistance and Biofilm Production in A Nepalese Hospital. Infection and Drug Resistance, 14: 2201-2212. DOI: https://doi.org/10.2147/IDR.S306688

Chow, J. W., Zervos, M. J., Lerner, S. A., Thal, L. A., Donabedian, S. M., Jaworski, D. D., Clewell, D. B. (1997): A Novel Gentamicin Resistance Gene in Enterococcus. Antimicrobial Agents and Chemotherapy 41: 511-514. DOI: https://doi.org/10.1128/AAC.41.3.511

Samani, R. J., Tajbakhsh, E., Momtaz, H., Samani, M. K. (2021): Prevalence of Virulence Genes and Antibiotic Resistance Pattern in Enterococcus faecalis Isolated from Urinary Tract Infection in Shahrekord, Iran. Reports of Biochemistry and Molecular Biology 10: 50. DOI: https://doi.org/10.52547/rbmb.10.1.50

Dutka-Malen, S., Evers, S., Courvalin, P. (1995): Detection of Glycopeptide Resistance Genotypes and Identification to the Species Level of Clinically Relevant Enterococci by PCR. Journal of Clinical Microbiology 33: 24-27. DOI: https://doi.org/10.1128/jcm.33.1.24-27.1995

Vergis, E. N., Shankar, N., Chow, J. W., Hayden, M. K., Snydman, D. R., Zervos, M. J., Muder, R. R. (2002): Association Between the Presence of Enterococcal Virulence Factors Gelatinase, Hemolysin, and Enterococcal Surface Protein and Mortality Among Patients with Bacteremia Due to Enterococcus faecalis. Clinical Infectious Diseases 35: 570-575. DOI: https://doi.org/10.1086/341977

Clinical and Laboratory Standards Institute (CLSI). (2023): Performance Standards for Antimicrobial Disk and Dilution Susceptibility Tests for Bacteria Isolated from Animals. 6th ed. CLSI supplement VET01S.

Zheng, J. X., Bai, B., Lin, Z. W., Pu, Z. Y., Yao, W. M., Chen, Z., Yu, Z. J. (2018): Characterization of Biofilm Formation by Enterococcus faecalis Isolates Derived from Urinary Tract Infections in China. Journal of Medical Microbiology 67: 60. DOI: https://doi.org/10.1099/jmm.0.000647

Cheng, W. N., Han, S. G. 2020: Bovine Mastitis: Risk Factors, Therapeutic Strategies, and Alternative Treatments-A Review. Asian-Australasian Journal of Animal Sciences 33: 1699. DOI: https://doi.org/10.5713/ajas.20.0156

Fiore, E., Van Tyne, D., Gilmore, M. S. (2019): Pathogenicity of Enterococci. Microbiology Spectrum 7: 7-4. DOI: https://doi.org/10.1128/microbiolspec.GPP3-0053-2018

Kudo, M., Nomura, T., Yomoda, S., Tanimoto, K., Tomita, H. (2014): Nosocomial Infection Caused by Vancomycin‐Susceptible Multidrug‐Resistant Enterococcus faecalis Over a Long Period in A University Hospital in Japan. Microbiology and Immunology 58: 607-614. DOI: https://doi.org/10.1111/1348-0421.12190

Yoon, S., Lee, Y. J. (2021): Molecular Characteristics of Enterococcus faecalis and Enterococcus faecium from Bulk Tank Milk in Korea. Animals 11: 661. DOI: https://doi.org/10.3390/ani11030661

Kim, H. J., Youn, H. Y., Kang, H. J., Moon, J. S., Jang, Y. S., Song, K. Y., Seo, K. H. (2022): Prevalence and Virulence Characteristics of Enterococcus faecalis and Enterococcus faecium in Bovine Mastitis Milk Compared to Bovine Normal Raw Milk in South Korea. Animals 12: 1407. DOI: https://doi.org/10.3390/ani12111407

Hanchi H., Mottawea W., Sebei K., Hammami R. (2018): The Genus Enterococcus: Between Probiotic Potential and Safety Concerns-An Update. Frontiers in Microbiology 9: 1791. DOI: https://doi.org/10.3389/fmicb.2018.01791

Gürler, H., Fındık, A., Gültiken, N., Ay, S. S., Çiftçi, A., Koldaş, E., Arslan, S., Fındık, M. (2015): Investigation on the Etiology of Subclinical Mastitis in Jersey and Hybrid Jersey Dairy Cows. Acta Veterinaria-Beograd, 65: 358-370. DOI: https://doi.org/10.1515/acve-2015-0030

Kuyucoğlu, Y. (2011): Sığır Subklinik Mastitisinden İzole Edilen Enterokokların Antibiyotik Dirençleri. Eurasian Journal of Veterinary Sciences 27: 231-234.

Herkmen, T. B., Türkyılmaz, S., (2016): Mastitisli Sığırlardan İzole Edilen Enterococcus faecium İzolatlarında GelE, Esp ve EfaAfm Genlerinin Varlığının İncelenmesi. Kocatepe Veterinary Journal 9: 54-60. DOI: https://doi.org/10.5578/kvj.20995

Cameron, M., Saab, M., Heider, L., McClure, J. T., Rodriguez-Lecompte, J. C., Sanchez, J. (2016): Antimicrobial Susceptibility Patterns of Environmental Streptococci Recovered from Bovine Milk Samples in the Maritime Provinces of Canada. Frontiers in Veterinary Science 3: 79. DOI: https://doi.org/10.3389/fvets.2016.00079

Cervinkova, D., Vlkova, H., Borodacova, I., Makovcova, J., Babak, V., Lorencova, A., Jaglic, Z. (2013): Prevalence of Mastitis Pathogens in Milk from Clinically Healthy Cows. Vet Med 58: 567-575. DOI: https://doi.org/10.17221/7138-VETMED

Kateete, D. P., Kabugo, U., Baluku, H., Nyakarahuka, L., Kyobe, S., Okee, M., Joloba, M. L. (2013): Prevalence and Antimicrobial Susceptibility Patterns of Bacteria from Milkmen and Cows with Clinical Mastitis in and Around Kampala, Uganda. Plos One 8: e63413. DOI: https://doi.org/10.1371/journal.pone.0063413

Hamzah, A. M., Kadim, H. K. (2018): Isolation and Identification of Enterococcus faecalis from Cow Milk Samples and Vaginal Swab from Human. Journal of Entomology and Zoology Studies 6: 218-222.

Cui, P., Feng, L., Zhang, L., He, J., An, T., Fu, X., Yang, X. (2020): Antimicrobial Resistance, Virulence Genes, and Biofilm Formation Capacity Among Enterococcus Species from Yaks in Aba Tibetan Autonomous Prefecture, China. Frontiers in Microbiology 12: 1250. DOI: https://doi.org/10.3389/fmicb.2020.01250

Lazzaro, L. M., Cassisi, M., Stefani, S., Campanile, F. (2022): Impact of PBP4 Alterations on Β-Lactam Resistance and Ceftobiprole Non-Susceptibility Among Enterococcus faecalis Clinical Isolates. Frontiers in Cellular and Infection Microbiology 11: 1441. DOI: https://doi.org/10.3389/fcimb.2021.816657

Amini, F., Krimpour, H. A., Ghaderi, M., Vaziri, S., Ferdowsi, S., Azizi, M., Amini, S. (2018): Prevalence of Aminoglycoside Resistance Genes in Enterococcus Strains in Kermanshah, Iran. Iranian Journal of Medical Sciences 43: 487.

Moussa, A. A., Md Nordin, A. F., Hamat, R. A., Jasni, A. S. (2019): High Level Aminoglycoside Resistance and Distribution of the Resistance Genes in Enterococcus faecalis and Enterococcus faecium from Teaching Hospital in Malaysia. Infection and Drug Resistance 12: 3269-3274. DOI: https://doi.org/10.2147/IDR.S219544

Taji, A., Heidari, H., Ebrahim-Saraie, H. S., Sarvari, J., Motamedifar, M. (2019): High Prevalence of Vancomycin and High-Level Gentamicin Resistance in Enterococcus faecalis Isolates. Acta Microbiologica et Immunologica Hungarica 66: 203-217. DOI: https://doi.org/10.1556/030.65.2018.046

Ahmed, M. O., Baptiste, K. E. (2018): Vancomycin-Resistant Enterococci: A Review of Antimicrobial Resistance Mechanisms and Perspectives of Human and Animal Health. Microbial Drug Resistance 24: 590-606. DOI: https://doi.org/10.1089/mdr.2017.0147

Quiñones, D., Aung, M. S., Martins, J. S., Urushibara, N., Kobayashi, N. (2018): Genetic Characteristics of VanA-Type Vancomycin-Resistant Enterococcus faecalis and Enterococcus faecium in Cuba. New Microbes and New Infections 21: 125-127. DOI: https://doi.org/10.1016/j.nmni.2017.12.001

Padmasini, E., Padmaraj, R., Ramesh, S. S. (2014): High Level Aminoglycoside Resistance and Distribution of Aminoglycoside Resistant Genes Among Clinical İsolates of Enterococcus Species in Chennai, India. The Scientific World Journal 329157. DOI: https://doi.org/10.1155/2014/329157

Li, W., Li, J., Wei, Q., Hu, Q., Lin, X., Chen, M., Lv, H. (2015): Characterization of Aminoglycoside Resistance and Virulence Genes Among Enterococcus sp. Isolated From A Hospital in China. International Journal of Environmental Research and Public Health 12: 3014-3025. DOI: https://doi.org/10.3390/ijerph120303014

Downloads

Published

2023-10-25

How to Cite

Özavci, V., Yüksel Dolgun, H. T., Seferoğlu, Y. ., & Kırkan, Şükrü. (2023). EVALUATION OF VIRULENCE AND ANTIMICROBIAL RESISTANCE GENES OF ENTEROCOCCUS SPECIES ISOLATED FROM SHEEP MILK WITH SUBCLINICAL MASTITIS. Journal of Applied Biological Sciences, 17(3), 467–479. https://doi.org/10.71336/jabs.1253

Issue

Section

Articles

Similar Articles

1 2 3 4 5 6 7 8 9 10 > >> 

You may also start an advanced similarity search for this article.