Abdali, M., Abdulrazzaq, A., Hamouda, R. (2023). Vancomycin-Resistant Enterococcus spp. Isolated From Mastitic cow’s milk. Journal of Applied Veterinary Sciences, 8(3), 8-15. doi: 10.21608/javs.2023.207770.1224
Abdali, M., Abdulrazzaq, A., Hamouda, R. (2023). 'Vancomycin-Resistant Enterococcus spp. Isolated From Mastitic cow’s milk', Journal of Applied Veterinary Sciences, 8(3), pp. 8-15. doi: 10.21608/javs.2023.207770.1224
Abdali, M., Abdulrazzaq, A., Hamouda, R. Vancomycin-Resistant Enterococcus spp. Isolated From Mastitic cow’s milk. Journal of Applied Veterinary Sciences, 2023; 8(3): 8-15. doi: 10.21608/javs.2023.207770.1224
Vancomycin-Resistant Enterococcus spp. Isolated From Mastitic cow’s milk
1Department of Microbiology, College of Veterinary Medicine, University of Baghdad, Baghdad, Iraq
2Head researcher at Animal Reproduction Research Institute, Agriculture Research Center, Giza, Egypt
Receive Date: 30 April 2023,
Revise Date: 01 June 2023,
Accept Date: 15 June 2023
Abstract
This study assessed the recrudescence of Enterococci in mastitis cow's milk and investigated their resistance to vancomycin. 300 samples were compiled from separate places and fields in Kirkuk, Iraq during the period from February to May, 2022. The samples were ioculated on the surface of bile esculin agar plates with sodium azide and then incubated at 37°C for 24–48 h. The characteristic pin-point colonies with a zone of black precipitate and morphologically resembling Enterococci spp. were further subjected to presumptive identification by Gram’ staining, catalase, and oxidase tests. All isolates were kept in BHIB with 30% glycerol at −70°C for further molecular detection. Enterococci isolates were tested for their susceptibility to different antibiotics by a disc diffusion technique. Based on the results of the sensitivity test, the ten isolates with the highest level of multiple resistances were selected from each of E. faecalisand E. faecium to examine the vanA, vanB genes by cPCR.The results of the bacteriological examination revealed that, 61 isolates (20.3%) of Enterococci According to phenotypic criteria; 42isolates were E. faecalis and 19 were E. faecium. Add this to the confirmatory tests that revealed 25 isolates (8.3%) were E. faecalis and 10 isolates (3.3%) were E. faecium detected by PCR. Antimicrobial susceptibility tests indicated high levels of multi-resistant E. faecalis and E. faecium strains. Vancomycin-resistant strains were 40% and 30% for E. faecalis and E. faecium, respectively. The genetic sequences of E. faecalis and E. faecium isolates and phylogenetic trees were established and registered in GenBank-NCBI. They obtained accession numbers (OP566382) for E. faecium and (OP566380) for E. faecalis, which became references in Iraq and around the world.
ABDEEN, E. E., HUSSIEN, H., HUSSAN, Z., and ABDELLA, W., 2016. Genotyping and virulence genes of Enterococcus faecalis isolated form kareish cheese and minced meat in Egypt. ResearchJournal of Microbiology, 11, 133-138.
ABDELTAWAB, A.W. 2019. Genotyping and resistance genes of Enterococcus faecalis isolated from different food sources in Egypt. Benha Vet Med J. 37 (1): 149-53. https://doi.org/10.21608/bvmj.2019.16026.1080
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. Int J Mol Cell Med. 4:240–4.
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. https://file.qums.ac.ir/repository.
CUI, P., FENG, L., ZHANG, .L, HE, J., AN, T., and FU, X., 2020. Antimicrobial resistance, virulence genes, and biofilm formation capacity among enterococcus species from yaks in Aba Tibetan Autonomous Prefecture, China. Front Microbiol. 11:1250.
DURAN, N., OZER, B., DURAN, G.G., ONLEN, Y., and DEMIR, C., 2012. Antibiotic resistance genes & susceptibility patterns in staphylococci. Indian J Med Res. 135(3):389-396. https://www.ncbi.nlm.nih.gov.
EL-ZAMKAN, M.A., and 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(11):e0259584.
EMANEINI, M., HOSSEINKHANI, F. and JABALAMELI, F., 2016. Prevalence of Vancomycin resistant Enterococcus in Iran: a systematic review and meta-analysis. Eur J Clin Microbiol Infect Dis. 35:1387–92. https://doi.org/10.1007/s10096-016-2702-0
FACKLAM, R.R., and COLLINS, M.D., 1989. Identification of Enterococcus species isolated from human infections by a conventional test scheme. J Clin Microbiol. 27(4):731-734.
HAMZAH, A.M., and KADIM, H.K., 2018. Isolation and identification of Enterococcus faecalis from cow milk samples and vaginal swab from human. Entomol Zool Sci. 6, 218–22. https://www.entomoljournal.com/archives.
HOMAN, W.L., TRIBE, D., POZNANSKI, S., Li, M., HOGG, G., SPALBURG, E., and WILLEMS, R.J., 2002. Multilocus sequence typing scheme for Enterococcus faecium. J Clin Microbiol. 40(6):1963-1971. https://doi.org/10.1128%2FJCM.40.6.1963-1971.2002
JACKSON, C.R., LOMBARD, J.E., DARGATZ, D.A., and FEDORKA-CRAY, P.J., 2011. Prevalence, species distribution and antimicrobial resistance of enterococci isolated from US dairy cattle. Lett Appl Microbiol. 52(1):41-8. https://doi.org/10.1111/j.1472-765x.2010.02964.x
KARIYAMA, R., MITSUHATA, R., CHOW, J.W., CLEWELL, D.B., and KUMON, H., 2000. Simple and reliable multiplex PCR assay for surveillance isolates of vancomycin-resistant enterococci. J Clin Microbiol. 38(8):3092-3095. https://doi.org/10.1128/jcm.38.8.3092-3095.2000
KIM, H.J., YOUN, H.Y., KANG, H.J., MOON, J.S., JANG, Y.S., SONG, K.Y., and 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(11): 1407. https://doi.org/10.3390/ani12111407
MOHAMMED, F., and AL-MARJAN, I., 2013. vanA in Vancomycin-resistant Enterococcus faecalis isolated in Baghdad. Afri J Microbiol Res. 7(2),: 115-9. http://dx.doi.org/10.5897/AJMR12.1443
MUNDY, L.M., SAHN, D.F., and GILMORE, M., 2000. Relationships between enterococcal virulence and antimicrobial resistance. Clin Microbiol Rev.13:513–522. https://doi.org/10.1128%2Fcmr.13.4.513-522.2000
NAM, H.M., LIM, S.K., MOON, J.S., KANG, H.M., KIM, J.M., JANG, K.C., KANG MI, JOO, Y.S., and JUNG SC., 2010. Antimicrobial resistance of enterococci isolated from mastitic bovine milk samples in Korea. Zoon Public Health. 57, 59-64.
ORABABA, O., and AKINSUYI, S.O., 2021. A systematic review and meta-analysis on the prevalence of Vancomycin-resistant enterococci (VRE) among Nigerians. Porto Biomed J. 6(1): e125. https://doi.org/10.1097%2Fj.pbj.0000000000000125
PATEL, R., UHL, J.R., KOHNER, P., and HOPKINS, M.K., 1997. Cockerill FR 3rd. Multiplex PCR detection of vanA, vanB, vanC-1, and vanC-2/3 genes in enterococci. J Clin Microbiol. 35(3):703-707. https://doi.org/10.1128%2Fjcm.35.3.703-707.1997
PESAVENTO, G., CALONICO, C., DUCCI, B., MAGNANINI, A., and LO NOSTRO, A., 2014. Prevalence and antibiotic resistance of Enterococcus spp. isolated from retail cheese, ready–to–eat salads, ham, and raw meat. Food Microbiol. 41: 1–7. DOI: https://doi.org/10.1016/j.fm.2014.01.008
RÓZANSKA, H., LEWTAK-PIŁAT, A., KUBAJKA, M., and WEINER, M., 2019. Occurrence of enterococci in mastitic cow’s milk and their antimicrobial ˙resistance. J. Vet. Res. 63(1):93–97. https://doi.org/10.2478/jvetres-2019-0014
SAMBROOK, J., FRITSCH, E.R., MANIATIS, T.. MOLECULAR, and CLONING, 1989. A Laboratory Manual (2nd ed.). Cold Spring Harbor, NY: Cold Spring Harbor Laboratory Press.,vol(1).
TEKIN, K., KADIR, S.G., and HARUN, H., 2016. Vancomycin Resistance of Enterococcus faecalis and Enterococcus faecium Isolated from cattle milk, Erciyes Üniv. Vet. Fak. Derg. 13(2) 139-150.
ZOLETTI, G.O., SIQUEIRA, J.F., and SANTOS, K.R.N., 2006. Identification of Enterococcus faecalis in root-filled teeth with or without periradicular lesions by culturedependent and -independent approaches. J Endodont.32(8): 231-9.