MOLECULAR CHARACTERIZATION OF MULTI-DRUG RESISTANCE ENTEROCOCCUS FAECALIS ISOLATED FROM MASTITIC COWS MILK

Authors

  • Marwa H. A.
  • A. Abdulrazzaq

DOI:

https://doi.org/10.36103/p6j22287

Keywords:

PCR, bacteria, gene, isolates, strains, contamination

Abstract

The study objects were to assess  Enterococcus faecalis  prevalence in milk from mastitis infected cows, and access their antibiotic resistance as well as investigate their resistance genes. A total of 300 samples were tested during the period from February to May in 2022. The results of initial examination showed that only 81 isolates as E. species according to phenotypic criteria. Confirmatory test was conducted, the results showed that only 25 isolates by 8.3% E.faecalis isolates. Antimicrobial susceptibility test indicated high levels of contamination with multi-drug resistant E.faecalis of with isolation of strains resistant to Vancomycin as 32%. The most high level of E.faecalis resistance was observed to Azithromycin, Cephalosporines, by 80 and 72% respectively. Adopt on results of susceptibility test, the six most isolates that gave high (MDR) were selected  to investigate resistance genes in them by PCR,  norA, tetK and aac6 genes are found in all of the isolate. E. faecalis gene was sequenced, analyzed, and registered in Gen-bank-NCBI and obtained the accession number (OP566380) that became a reference in Iraq and the world.

References

1. Ashraf A. A. 2019. Genotyping and resistance genes of Enterococcus faecalis isolated from different food sources in Egypt. Benha University. Benha Veterinary Medical Journal. September 2019

DOI: 10.21608/bvmj.2019.16026.1080.

2. Atiyah, A.S. and M.H. Alkhafaji.2020. Isolation of Enterococcus Species from Food Sources and Its Antibacterial Activity against Staphylococcus Aureus, Iraqi Journal of Sciences. 61(12): 3164-3171.

DOI: 10.24996/ijs.2020.61.12.3.

3. Bagcigil, A.B., S.Taponen, J. Koort, B.Bengtsson, A Myllyniemi and S.Pyörälä.2012. Genetic basis of penicillin resistance of S. aureus isolated in bovine mastitis. Acta Veterinaria Scandinavica. 54:69.

4. CLSI- M100 2018. Performance Standard for Antimicrobial Susceptibility Testing-Table (2D): Zone Diameter and MIC Breakpoints for Enterococcus spp., Clinical and Laboratory Standards Institute, 28th Edition. 38 (3): 64–67.

5. Duran, N., B.Ozer, G.G. Duran, Y.Onlen and C.Demir. 2012. Antibiotic resistance genes and susceptibility patterns in staphylococci. Indian J. Med. Res. 135: 389-396.

6. Espeche, M. C., Pellegrino, M., Frola, I., Larriestra, A., Bogni, C., & Nader-Macías, M. F. 2012. Lactic acid bacteria from raw milk as potentially beneficial strains to prevent bovine mastitis. Anaerobe, 18(1), 103-109. https://doi.org/10.1016/j.anaerobe.2012.01.002

7. Erbas, G., Parın, U., Turkyılmaz, S., Ucan, N., Ozturk, M., & Kaya, O. 2016. Distribution of antibiotic resistance genes in Enterococcus spp. isolated from mastitis bovine milk.

DOI: 10.1515/acve-2016-0029

8. Facklam, R.R. and M.D. Collins.1989. Identification of Enterococcus species isolated from human infections by a conventional test scheme. J. Clin. Microbiol. 27: 731-734.

9. Ghaidaa, J. M. 2022. Molecular genotyping of Enterococcus faecalis isolated from clinical sources by Box-PCR and Eric-PCR. University of Al-Qadisiyah, International Journal of Health Sciences·

DOI: 10.53730/ijhs.v6nS1.7857.

10. Hamzah, A.M. and H.K. Kadim 2018. Isolation and identification of Enterococcus faecalis from cow milk samples and vaginal swab from human. Entomol Zool Sci. 6:218–222.

11. Kariyama, R.,R. Mitsuhata, J. W. Chow, D.B. Clewell and H. Kumon. 2000. Simple and reliable multiplex PCR assay for surveillance isolates of vancomycin-resistant enterococci. J. Clinic.Microbiol.3092–3095.

12. Kehrenberg, C. and S. Schwarz. 2006. Distribution of florfenicol resistance genes fexA and cfr among chloramphenicol-resistant Staphylococcus isolates. Antimicrob.Agets and Chomether. Apr. 2006, P: 1156–1163.

13. Kim, H. J., H. Y. Youn, H.J. Kang, J.S. Moon, K.Y. Jang and, K.H.Song.2002. 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.

https://doi.org/10.3390/ ani12111407.

14. Kuyucouğlu ,Y. 2011. Antibiotic resistances of enterococci isolated from bovine subclinical mastitis. Eurasian. J .Vet. Sci. 27:231–234.

15. Nam, H. M., Lim, S. K., Moon, J. S., Kang, H. M., Kim, J. M., Jang, K. C., ... and Jung, S. C. 2010. Antimicrobial resistance of enterococci isolated from mastitic bovine milk samples in Korea. Zoonoses and public health, 57(7‐8), e59-e64. https://doi.org/10.1111/j.1863-2378.2009.01307.x

16. Nam, H.M., S.K. Lim, J.S. Moon, H.M. Kang, J.M. Kim, K.C. Jang, J.M.Kim, M.I. Kang, Y.S. Joo and S.C. Jung.2010. Antimicrobial Resistance of Enterococci Isolated from Mastitic Bovine Milk Samples in Korea. Zoonoses Public Health. 57, pp: 59–e64.

17. Patel, R., J.R. P.Uhl, M.K. Kohner and F.R. Cockerill.1997. Multiplex PCR detection of vanA, vanB, vanC-1, and vanC-2/3 genes in Enterococci. J. Clinic. Microbiol. 35(3):703-707.

18. Pourmand, M.R., M.Yousefi, S.A. Salami and M.Amini.2014. Evaluation of expression of norA efflux pump in ciprofloxacin resistant Staphylococcus aureus against hexahydroquinoline derivative by Real-Time PCR. Acta Med. Iran.52(6):424-9.

19. Róza ´nska, H., A.Lewtak-Piłat, M.Kubajka and M.Weiner.2019. Occurrence of enterococci in mastitic cow’s milk and their antimicrobial ˙resistance. J. Vet. Res.63: 93–97.

20. Rowe, S. M., S. M. Godden, E. Royster, J. Timmerman, and M. Boyle, 2021. Postcalving udder health and productivity in cows approaching dry-off with intramammary infections caused by non-aureus Staphylococcus, Aerococcus, Enterococcus, Lactococcus, and Streptococcus species. Journal of Dairy Science, 104(5), 6061-6079.

https://doi.org/10.3168/jds.2020-19288

21. Sambrook, J., E.F.Fritscgh, and Mentiates T. 1989. Molecular Coloning. A Laboratory Manual. V. 3. Cold Spring Harbor Laboratotry Press, New York. vol(1). https://www.scirp.org.

22. Schlegelova, J., H.Vlkova, V.Babak, M.Holasova, Z. Jaglic, T.Stosova and P. Sauer.2008.Resistance to erythromycin of Staphylococcus spp. isolates from the food chain. Veterinarni Medicina. 53 (6): 307–314.

23. Sumaya, Y.A. Bacterial Bovine Mastitis in Iraq. College of Veterinary Medicine . Bas.J.Vet. Res.19 (2) 2020 .

24. Van Tyne D. and M.S. Gilmore. 2014. Friend turned foe: Evolution of enterococcal virulence and antibiotic resistance, Annual Review of Microbiology. 68: 337–356.

25. Zoletti, G.O., J.F. Siqueira and K.R.N. Santos. Identification of Enterococcus faecalis in root-filled Teeth with or without periradicular lesions by culture dependent and independent Approaches. J.O.E. 32(8) 2006.

Downloads

Published

2025-06-28

Issue

Section

Articles

How to Cite

Marwa H. A., & A. Abdulrazzaq. (2025). MOLECULAR CHARACTERIZATION OF MULTI-DRUG RESISTANCE ENTEROCOCCUS FAECALIS ISOLATED FROM MASTITIC COWS MILK. IRAQI JOURNAL OF AGRICULTURAL SCIENCES, 56(3), 1021-1028. https://doi.org/10.36103/p6j22287