DETECTION of SOME VIRULENCE FACTORS and ANTIBIOTIC SENSITIVITY TEST of ENTEROCOCCUS FAECALIS ISOLATED FROM SHEEP by MULTIPLEX PCR
European Journal of Molecular & Clinical Medicine,
2020, Volume 7, Issue 9, Pages 68-74
AbstractEnterococcus faecalis form an important population of commensal bacteria and have been reported to possess numerous virulence factors considered significantly important in exacerbating diseases caused by them. Objectives: The present study was conducted to evaluates the presence of virulence factors and antibiotic susceptibility among Enterococcus faecalis isolated from sheep. Methods: The study included the collection of 50 samples (25 Milk samples collected from the udder was washed and the teats were disinfected and dried using alcohol, the first milk drop removed. 5ml of milk collected on aseptic tube and 25 Feces samples collected from sheep diarrhea from rectal by aseptic gloves. (from October 2018 to March 2019 ) and transported to laboratory as soon as possible in sterile Brain heart infusion broth that incubated at 37 C for at least 24-28 hours to increasing chances of isolation. Enterococcus faecalis that were recognized by cultural characteristics, Gram stain, and biochemical reactions. Results: The results of the laboratory cultural of 50 cotton swabs used s show that the isolation rate of Enterococcus spp. were 32% and 56% from milk and feaces respectively. the result of PCR test for detection of Enterococcus faecalis: show that the Enterococcus faecalis detected in rate of 66.6% from total Enterococcus spp. While the result of Enterococcus faecalis virulence factors showed that the Surface proteins, Gelatinase and Hemolysin were 75%, 33.3%, 25.5% respectively. Results of antibiotic sensitivity test showed the most bacterial isolated sensitive Nitrofurantoin , Imipenem and Nalidixic acid were 91.6%,83.3% and 58.3% % respectively Conclusion: We report that our simple modification of the existing multiplex PCR had increased the detection of the enterococcal virulence genes. Predominance of virulence genes was in order of Surface proteins, Gelatinase and Hemolysin were 75%, 33.3%, 25.5%. This modified PCR protocol could be useful to resolve the problem of decreased detection of virulence determinants in enterococci.
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