Study of virulence genes in vancomycin resistant Enterococci (vre) from animals and human clinical isolates

Paper Details

Research Paper 01/01/2021
Views (350) Download (41)

Study of virulence genes in vancomycin resistant Enterococci (vre) from animals and human clinical isolates

Mofoluwaso Adedeji Oyinloye, Bondira Olamide Ndagana, Adeola Egbebi, Pius Abimbola Okiki
Int. J. Biosci.18( 1), 1-14, January 2021.
Certificate: IJB 2021 [Generate Certificate]


With Enterococcus species in the leading cause of nosocomial infections and resistance to an array of antibiotics, this study focused to determine the frequency and distribution of vancomycin-resistant Enterococci, the presence of virulence genes and to determine the relative nucleotide sequence relatedness among isolates using 16S rRNA sequence. A random sampling of 120 fecal samples of cattle, poultry, and piggery, and human clinical isolates was analyzed. Standard bacteriological methods were employed in the isolation and characterization of isolates and the disk diffusion method was used in determining their antibiotic resistance profiles. Results showed Enterococcus species in cattle at 100%, followed by clinical isolates at 80%. Vancomycin resistance was observed at high rates in Enterococcus species from human clinical isolates and cattle isolates at 90% and 80% respectively. Multiple antibiotic-resistant isolates yielded twelve resistance profiles and 16S rDNA sequences identified E. faecalis, E. durans, E. mundtii, and Enterococcus sp. Isolates from cattle samples were the most probable source of clinical isolates at 78% homology of conserved regions with the clinical isolates.  Virulence determinant genes Asa1 was recorded at66.6%, Cyl at 16.6% and GelE at 8.3% among the isolates. This study established farm animals as possible reservoirs of VRE isolates to man. Hence, healthy and professional practices among animal farmers with antibiotic usage, as well as hygienic and preventive measures among hospital workers are here recommended.


Ali SA, Hasan KA, Bin Asif H, Abbasi A. 2014. Environmental Enterococci: Prevalence of Virulence, Antibiotic Resistance and Species Distribution in Poultry and its Related Environment in Karachi, Pakistan. Letters in Applied Microbiology 58(5), 423–432.

Barlow RS, McMillan KE, Duffy LL, Fegan N, Jordan D, Mellor GE. 2017. Antimicrobial Resistance Status of Enterococcus from Australian Cattle Populations at Slaughter. PLoS One 12(5), 1–13.

Bonten MJ, Willems RJ. 2012. Vancomycin-resistant Enterococcus chronicle of a foretold problem. Nederlands Tijdschriftvoor Geneeskunde 156, 52-33.

Byappanahalli MN, Nevers MB, Korajkic A, Staley ZR, Harwood VJ. 2012. Enterococci in the Environment. Microbiology and Molecular Biology Reviews 76(4), 685–706.

Cattoir V, Giard JC. 2014. Antibiotic Resistance in Enterococcus faecium Clinical Isolates. Expert Review of Anti-Infective Therapy 12, 239–248.

Cercenado E, Ûnal S, Eliopoulos CT, Rubin LG, Isenberg HD, Moellering RC, Eliopoulos GM. 1995. Characterization of vancomycin resistance in Enterococcus durans. Journal of Antimicrobial Chemotherapy 36, 821–5.

Clinical, Laboratory Standard Institute. 2016. Performance Standards for Antimicrobial Susceptibility Testing: Test Approved Standard. CLSI document no M100-S26.

Collins MD, Farrow JA, Jones D. 1986. Enterococcus mundtii sp. nov.International Journal of Systematic Evolutionary Microbiology 36, 8–12.

Comerlato CB, Resende MC, Caierao J, d’Azevedo PA. 2013. Presence of virulence factors in Enterococcus faecalis and Enterococcus faecium susceptible and resistant to vancomycin. Memórias do InstitutoOswaldo Cruz 108(5), 590–595.

De Kwaadsteniet M, Todorov SD, Knoetze H, Dicks LM. 2005. Characterization of a 3944 Da bacteriocin, produced by Enterococcus mundtii ST15, with activity against Gram-positive and Gram-negative bacteria.International Journal of Food Microbiology 105, 433–444.

Descheemaeker P, Ieven M, Chappelle S, Lammens C, Hauchecorne M, Wijdooghe M, Vandamme P, Goossens H. 2000. Prevalence and molecular epidemiology of glycopeptide-resistant enterococci in Belgian renal dialysis units.Journal of Infectious Diseases 181, 235–41.

Diego C, Andrigetto C, Lombardi A. 2016. Occurrence of virulence factors and Antibiotic Resistance in Enterococcus faecalis and Enterococcus faecium Collected From Dairy and Human Samples in North Italy. Food Control 19(9), 886-892.

Espeche MC, Otero MC, Sesma F, Nader-Macias ME. 2009. Screening of surface properties and antagonistic substances production by lactic acid bacteria isolated from the mammary gland of healthy and mastitic cows. Veterinary Microbiology 135, 346–357.

Facklam RR, Carvalho MG, Teixeira LM. 2002. Enterococcus. In M.S. Gilmore, Clewell D.B., Courvalin P., Dunny G.M., Murray B.E. and Rice L.B. The Enterococci: Pathogenesis, Molecular Biology, and Antibiotic Resistance. 1-54.

Fallavollita L, Di Gioacchino L, Balestrini F. 2016. Bioprosthetic aortic valve endocarditis in association with Enterococcus durans.Texas Heart Institute Journal 43(2), 165–167.

Ferguson DM, Talavera GN, Hernández LA, Weisberg SB, Ambrose RF, Jay JA. 2016. Virulence genes among Enterococcus faecalis and Enterococcus faecium isolated from coastal beaches and human and nonhuman sources in Southern California and Puerto Rico. Journal of Pathogenesis 2016, 3437214.

Ferreira AE, Canal N, Morales D, Fuentefria DB, Corção G. 2007. Characterization of enterocins produced by Enterococcus mundtii isolated from humans feces. Brazilian Archives of Biology and Technology 50, 249–258.

Flokas ME, Karageorgos SA, Detsis M, Alevizakos M, Mylonakis E. 2017.Vancomycin-Resistant Enterococci Colonisation, Risk Factors and Risk for Infection Among Hospitalized Paediatric Patients: a Systematic Review and Metanalysis. International Journal of Antimicrobial Agents 49, 565-572.

Foka FET, Ateba CN. 2019. Detection of Virulence Genes in Multidrug Resistant Enterococci Isolated from Feedlots Dairy and Beef Cattle: Implications for Human Health and Food Safety. BioMedical Research International, Article ID 5921840, 13 pages.

Giraffa G. 2003. Functionality of enterococci in dairy products. International Journal of Food Microbiology 88, 215–222.

Giraffa G, Carminati D, Neviani E. 1997. Enterococci isolated from dairy products: a review of risks and potential technological use. Journal of Food Protection 60, 732–738.

Hall LM, Chen HY Williams RJ. 1992. Vancomycin-resistant Enterococcus durans. Lancet 340, 1105.

Higashide T, Takahashi M, Kobayashi A, Ohkubo S, Sakurai M, Shirao Y, Tamura T, Sugiyama K. 2005. Endophthalmitis Caused by Enterococcus mundtii. Journal of Clinical Microbiology 43(3), 1475–1476.

Kenzaka T, Takamura N, Kumabe A, Takeda K. 2013. A case of subacute infective endocarditis and blood access infection caused by Enterococcus durans. BMC Infectious Diseases 13, 594.

Kondoh M, Fukada K, Fujikura D, Shimada T, Suzuki Y, Iwai A. 2012. Effect of Water-Soluble Fraction from Lysozyme-Treated Enterococcus faecalis FK-23 on Mortality Caused by Influenza A virus in Mice. Viral Immunology 25(1), 86–90.

Lebreton F, Depardieu F, Bourdon N, Fines-Guyon M, Berger P, Camiade S. 2011. D- Ala-d-SerVanN-type Transferable Vancomycin Resistance in Enterococcus faecium. Antimicrobial Agents Chemotherapy 55, 4606–4612.

Liliana L, Diana M, Bessa L, Angelo M, Augusto J, DeMatos J, Paulo Martins da Costa. 2014. Spread of Multidrug-Resistant Enterococcus faecalis Within the Household Setting. Microbial drug resistance 20, 5-7.

Meyer K, Schonfeld H. 1926. Über die Unterscheidung des Enterococcus von Streptococcus viridans und die BeziehungbeiderzumStreptococcus lactis. Zentralbl. Bakteriol. Parasitenkol. Infektionskr. Hyg. Abt. 1 Orig. 99, 402–416.

Moraes PM, Perin LM, Todorov SD, Silva A, Franco BD, Nero LA. 2012. Bacteriocinogenic and Virulence Potential of Enterococcus Isolates Obtained from raw Milk and Cheese. Journal of Applied Microbiology 113(2), 318–328.

Naser S, Thompson FL, Hoste B, Gevers D, Vandemeulebroecke K, Cleenwerck I, Thompson CC, Vancanneyt M, Swings J. 2005. Phylogeny and Identification of   Enterococci by atpA Gene Sequence Analysis. Journal of Clinical Microbiology 43(5), 2224–2230.

Olawale AK, Onasanya A, Oyelakin OO, David OM, Famurewa O. 2015. Enterococcus faecalis isolates of Food Origin and Detection of Their Virulence Determinant Genes in Osun State, Nigeria. Microbiology Research International 2(2), 18-27.

Ryu B, Hong J, Jung J, Min JK, Heungsup S, Mi-Na K, Yong PC, Sung-Han K, Sang-Oh L, Yang SK, Jun HW, Sang-Ho C. 2019. Clinical characteristics and treatment outcomes of Enterococcus durans bacteremia: a 20-year experience in a tertiary care hospital. European Journal of Clinical Microbiology and Infectious Diseases 38, 1743–1751.

Settanni L, Valmorri S, Suzzi G, Corsetti A. 2008. The role of environmental factors and medium composition on bacteriocin-like inhibitory substances (BLIS) production by Enterococcus mundtii strains.Food Microbiology 25, 722–728.

Stepanovic S, Jovanovic M, Lavadinovic L, Stosovic B, Pelemis M. 2004. Enterococcus durans endocarditis in a patient with transposition of the great vessels. Journal of Medical Microbiology 53(3), 259–261.

Trivedi K, Cupakova S, Karpiskova R. 2011. Virulence factors and antibiotic resistance in enterococci isolated from food-stuffs. Veterinarni Medicina 56(7), 352–357.

Upadhyaya GPM, Ravikumar KL, Umapathy BL. 2009. “Review of virulence factors of Enterococcus: an emerging nosocomial pathogen,” Indian Journal of Medical Microbiology 27(4), 301–305.

Van Harten RM, Willems RJL, Martin NI, Hendrickx APA. 2017. Multidrug-Resistant Enterococcal Infections: New Compounds, Novel Antimicrobial therapies. Trends in Microbiology 25, 467–479.

Vijayakrishnan R, Rapose A. 2012. Fatal Enterococcus durans aortic valve endocarditis: a case report and review of the literature. BMJ Case Report.

Yilema A, Moges FS, Tadele LL. 2017. Isolation of Enterococci, their antimicrobial susceptibility patterns and associated factors among patients attending at the University of Gondar Teaching Hospital. BMC Infectious Diseases 17, 276.

Yılmaz EŞ, Aslantaş O, Onen SP, Turkyilmaz S, Kurekci C. 2016. Prevalence, Antimicrobial Resistance and Virulence Traits in Enterococci from food of animal origin in Turkey. Food Science and Technology 66, 20–26.

Zala A, Collins N. 2016. Enterococcus durans prosthetic valve endocarditis: a previously unreported clinical entity. Heart, Lung and Circulation 25(10), e133–e136.