Chaitanya Gottapu¹*, Ch. Bindu Kiranmayi², T. Srinivasa Rao³, K. Aswani Kumar⁴, N. Subashini⁵, B. Swathi vimala⁶, Srinivas K⁷, Suresh Yasarla⁸, Chaitanya Venkata⁶, B. Bhavana ⁶

¹Doctoral student, College of Veterinary Sciences, University of Florida ²Principal, Veterinary Polytechnic college, V.R. Gudem, Sri Venkateswara Veterinary University, Andhra Pradesh, India ³Professor and Head, Department of Veterinary Public Health and Epidemiology, NTR College of Veterinary Science, Gannavaram, Andhra Pradesh, India ⁴Professor and Head, Department of Veterinary Biochemistry, NTR College of Veterinary Science, Gannavaram, Andhra Pradesh, India ⁵Assistant professor, Department of Veterinary Public Health and Epidemiology, NTR College of Veterinary Science, Gannavaram, Andhra Pradesh, India ⁶M.V.Sc. Scholar, NTR College of Veterinary Science, Gannavaram, Andhra Pradesh, India ⁷Ph. D scholar, ICAR – Indian Veterinary Research Institute, Izatnagar, Uttar Pradesh, India ⁸Assistant professor, College of Veterinary Science, Tirupati, Andhra Pradesh, India

*Corresponding Author:Chaitanya Gottapu, Doctoral Student, College of Veterinary Sciences, University of Florida, Florida.

Received: July 18, 2022; Published: July 25, 2022

Abstract

Enterococci are the common opportunistic pathogens having a worldwide food safety concern. The present study was undertaken to characterize the vancomycin resistant Enterococcus species of animal, human and environmental origin by using ERIC-PCR and REP-PCR assays. Out of 608 Enterococcus spp. isolates recovered by phenotypic and genotypic methods, 125 Enterococcus isolates were identified as Vancomycin resistance Enterococcus genotypically. The vancomycin resistant genes vanB, vanC1 and vanC2 were detected in 14 (11.20%), 69 (55.20%) and 42 (36.60%) Enterococcus isolates, respectively. A greater degree of heterogeneity was observed among 124 VRE isolates (one E. gallinarum isolate did not yield any bands for both ERIC-PCR and REP- PCR) of four species of Enterococcus from different sources as revealed by presence of 122 genotypes and 123 genotypes by ERIC and REP-PCR analysis, respectively. Nineteen different E. faecalis, 15 E. faecium, 57 E. gallinarum and 31 E. casseliflavus subtypes were differentiated by ERIC-PCR, whereas 21 different E. faecalis, 15 E. faecium, 56 E. gallinarum and 31 E. casseliflavus subtypes by REP- PCR. Genotyping of VRE species by ERIC- PCR and REP- PCR were found to be highly significant since discriminatory power > 0.9 are considered highly significant (0.9997 for ERIC-PCR and 0.9999 for REP-PCR). Cluster analysis also revealed a great degree of homogeneity among some VRE isolates recovered from different sources and implied at the chance of cross-contamination of foods of animal origin.

Keywords: Enterococci; VRE; ERIC-PCR; REP-PCR; Discriminatory Power; Cluster Analysis

References

1. Arthur M., et al. “Glycopeptide resistance in enterococci”. Trends in Microbiology 10 (1996): 401-407.
2. Aarestrup FM. “Occurrence of glycopeptide resistance among Enterococcus faecium isolates from conventional and ecological poultry farms”. Microbial Drug Resistance 1 (1995): 255-257.
3. Klare I., et al. “VanA-mediated high-level glycopeptide resistance in Enterococcus faecium from animal husbandry”. FEMS Microbiology Letters 125 (1995): 165-171.
4. Kruse H., et al. “The use of avoparcine as a growth promoter and the occurrence of vancomycin-resistant Enterococcus species in Norwegian poultry and swine production”. Microbial Drug Resistance 5 (1999): 135-139.
5. Wegener HC., et al. “Use of antimicrobial growth promoters in food animals and Enterococcus faecium resistance to therapeutic antimicrobial drugs in Europe”. Emerging Infectious Diseases 5 (1999): 329-335.
6. Stobberingh E., et al. “Enterococci with glycopeptide resistance in turkeys, turkey farmers, turkey slaughterers, and (sub) urban residents in the south of the Netherlands: evidence for transmission of vancomycin resistance from animals to humans”. Antimicrobial Agents and Chemotherapy 43 (1999): 2215-2221.
7. Leclercq R., et al. “Plasmid-mediated resistance to vancomycin and teicoplanin in Enterococcus faecium”. The New England Journal of Medicine 3 (1988): 157-161.
8. Patel R. “Epidemiology and Mechanisms of Glycopeptide Resistance in Enterococci”. International Symposium on Antimicrobial Agents and Resistance. (5^th edition). ISAAR, Seoul, South Korea (2005): 40-43.
9. Taneja N., et al. “Epidemiology of vancomycin- resistant enterococci”. In: Kobayashi N Drug resistance of enterococci: epidemiology and molecular mechanisms. Research signpost, Trivandrum, Kerala, India (2006a): 79-99.
10. Blanco AE., et al. “Characterization of Enterococcus faecalis isolates by chicken embryo lethality assay and ERIC-PCR”. Avian Pathology1 (2018): 23-32.
11. Bedendo J and Pignatari ACC. “Typing of Enterococcus faecium by polymerase chain reaction and pulsed field gel electrophoresis”. Brazilian Journal of Medical and Biological Research11 (2000): 1269-1274.
12. Mendez-Alvarez S., et al. “Analysis of bacterial genomes by pulsed field gel electrophoresis”. Microbiologia (Madrid, Spain)3 (1995): 323-336.
13. Tenover FC. “Laboratory methods for surveillance of vancomycin-resistant enterococci”. Clinical Microbiology Newsletter1 (1998): 1-5.
14. Monstein HJ., et al. “Division of the genus Enterococcus into species groups using PCR-based molecular typing methods”. Microbiology5 (1998): 1171-1179.
15. Ke D., et al. “Development of a PCR assay for rapid detection of enterococci”. Journal of Clinical Microbiology 11 (1999): 3497-3503.
16. Scheidegger EMD., et al. “RFLP analysis of a PCR-amplified fragment of the 16S rRNA gene as a tool to identify Enterococcus strains”. Memórias do Instituto Oswaldo Cruz7 (2009): 1003-1008.
17. Maiden MC., et al. “Multilocus sequence typing: a portable approach to the identification of clones within populations of pathogenic microorganisms”. Proceedings of the National Academy of Sciences6 (1998): 3140-3145.
18. Enright MC and Spratt BG. “Multilocus sequence typing”. Trends in Microbiology12 (1999): 482-487.
19. Enright MC., et al. “Multilocus sequence typing for characterization of methicillin-resistant and methicillin-susceptible clones ofStaphylococcus aureus”. Journal of Clinical Microbiology3 (2000): 1008-1015.
20. Kalia A., et al. “Directional gene movement from human-pathogenic to commensal-like streptococci”. Infection and Immunity 69 (8 (2001): 4858-4869.
21. Savelkoul PHM., et al. “Amplified-fragment length polymorphism analysis: the state of an art”. Journal of Clinical Microbiology10 (1999): 3083-3091.
22. Antonishyn NA., et al. “Evaluation of fluorescence-based amplified fragment length polymorphism analysis for molecular typing in hospital epidemiology: comparison with pulsed-field gel electrophoresis for typing strains of vancomycin-resistant Enterococcus faecium”. Journal of Clinical Microbiology11 (2000): 4058-4065.
23. Behzadi P., et al. “Nucleic acid-based approaches for detection of viral hepatitis”. Jundishapur Journal of Microbiology1 (2015).
24. Ranjbar R., et al. “The study of genetic relationship among third generation cephalosporin-resistant Salmonella enterica strains by ERIC-PCR”. The Open Microbiology Journal 7 (2013a):
25. Ranjbar R., et al. “Molecular typing of Salmonella enteritidis strains isolated in several laboratory centers in Tehran by ERIC-PCR”. Scientific Journal of Kurdistan University of Medical Sciences2 (2013b).
26. Ranjbar R., et al. “Typing methods used in the molecular epidemiology of microbial pathogens: a how-to guide”. The New Microbiologica1 (2014): 1-15.
27. Hiett KL and Seal BS. “Use of repetitive element palindromic PCR (rep-PCR) for the epidemiologic discrimination of foodborne pathogens”. In Molecular Epidemiology of Microorganisms. Humana Press, Totowa, NJ (2009): 49-58.
28. Pavia M., et al. “Vancomycin resistance and antibiotic susceptibility of enterococci in raw meat”. Journal of Food Protection 63 (2000): 912-915.
29. Gambarotto K., et al. “Occurrence of vancomycin-resistant enterococci in pork and poultry products from a cattle-rearing area of France”. Journal of Clinical Microbiology39.6 (2001): 2354-2355.
30. Power EG., et al. “VanA genes in vancomycin-resistant clinical isolates of Oerskovia tura and Arcanobacterium (Corynebacterium) haemolyticum”. Journal of Antimicrobial Chemotherapy 4 (1995): 595-606.
31. Biavasco F., et al. “In vitro conjugative transfer of VanA vancomycin resistance between Enterococci and Listeriae of different species”. European Journal of Clinical Microbiology and Infectious Diseases 1 (1996): 50-59.
32. Franke AE and Clewell DB. “Evidence for a chromosome-borne resistance transposon (Tn916) in Streptococcus faecalis that is capable of "conjugal" transfer in the absence of a conjugative plasmid”. Journal of Bacteriology 145.1 (1981): 494-502.
33. Billot-Klein D., et al. “Modification of peptidoglycan precursors is a common feature of the low-level vancomycin-resistant VANB-type Enterococcus D366 and of the naturally glycopeptide-resistant species Lactobacillus casei, Pediococcus pentosaceus, Leuconostoc mesenteroides, and Enterococcus gallinarum”. Journal of Bacteriology 8 (1994): 2398-2405.
34. Klare I and Witte W. “Glycopeptide resistent Enterococcus: zur Situation in Deutschland”. Hygiene Microbiology 1.31-38 (1997):
35. Felsenstein J. “PHYLIP - phylogeny inference package” (version 3.6) cladistics 5 (1989): 164-166.
36. Hunter PR and Gaston MA. “Numerical index of the discriminatory ability of typing systems: an application of Simpson's index of diversity”. Journal of Clinical Microbiology11 (1988): 2465-2466.
37. Abadía-Patiño L., et al. “VanE-type vancomycin-resistant Enterococcus faecalis clinical isolates from Australia”. Antimicrobial Agents and Chemotherapy 48.12 (2004): 4882-4885.
38. Xavier DB., et al. “Absence of VanA and VanB containing enterococci in poultry raised on nonintensive production farms in Brazil”. Applied and Environmental Microbiology 72.4 (2006): 3072-3073.
39. Nishiyama M., et al. “Identification of Enterococcus faecium and Enterococcus faecalis as vanC-type vancomycin-resistant enterococci (VRE) from sewage and river water in the provincial city of Miyazaki, Japan”. Journal of Environmental Science and Health, Part A 50.1 (2015): 16-25.
40. Latha P., et al. “Multiplex PCR based genotypic characterization of pathogenic vancvancomycin-resistanterococcus faecalis recovered from an Indian river along a city landscape”. Springerplus 5.1 (2016):
41. Mac K., et al. “Species identification and detection of vancomycin resistance genes in enterococci of animal origin by multiplex PCR”. International Journal of Food Microbiology 88.2-3 (2003): 305-309.
42. Schwaiger K., et al. “Presence of the resistance genes vanC1 and pbp5 in phenotypically vancomycin and ampicillin susceptible Enterococcus faecalis”. Microbial Drug Resistance 18.4 (2012): 434-439.
43. Moura TMD., et al. “Detection of vanC1 gene transcription in vancomycin-susceptible Enterococcus faecalis”. Memórias do Instituto Oswaldo Cruz 108.4 (2013): 453-456.

Citation

Citation: Chaitanya Gottapu., et al. “Genetic Diversity of Vancomycin Resistance Enterococcus Spp. Isolated from Animal, Human and Environmental Origin". Acta Scientific Veterinary Sciences 4.8 (2022): 130-146.

Copyright

Copyright: © 2022 Chaitanya Gottapu., et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.