Environmental Science and Pollution Research

, Volume 26, Issue 15, pp 15105–15114 | Cite as

Virulence profiles of vancomycin-resistant enterococci isolated from surface and ground water utilized by humans in the North West Province, South Africa: a public health perspective

  • Daniel Pheeha Matlou
  • Marie Ebob AgborTabot Bissong
  • Christ-Donald Kaptchouang Tchatchouang
  • Mohomud Rashid Adem
  • Frank Eric Tatsing Foka
  • Ajay Kumar
  • Collins Njie AtebaEmail author
Research Article


Vancomycin-resistant enterococci (VRE) have been responsible for numerous outbreaks of serious infections in humans worldwide. Enterococcus faecium and Enterococcus faecalis are the principal species that are frequently associated with vancomycin resistance determinants, thus usually implicated in hospital- and community-acquired infections in humans. The study aim was to determine the antibiotic resistance and virulence profiles of VREs isolated from surface and groundwater samples that are used by humans in the North West Province, South Africa. A total of 170 water samples were collected and analyzed. Eighty-one potential isolates were screened for characteristics of Enterococcus species using preliminary biochemical tests, PCR assays and sequence analysis. The antimicrobial resistance profiles of the isolates against nine antibiotics were determined and a dendrogram was generated to access the relatedness of the isolates. The isolates were screened for the presence of antibiotic resistance and virulence genes by multiplex PCR analysis. A total of 56 isolates were confirmed as Enterococcus species and the proportion of E. faecium (46.9%) was higher than E. faecalis (29%) and E. saccharolyticus (1.2%). Sequence data of E. faecium, E. faecalis, and E. saccharolyticus isolates revealed 97 to 98% similarities to clinical strains deposited in NCBI Genbank. Large proportions (44; 78.6%) of the isolates were resistant to vancomycin while 16 and 3.6% of the isolates possessed the vanA and vanB genes respectively. The MAR phenotype Vancomycin-Nalidixic Acid-Streptomycin-Chloramphenicol-Ampicillin-Oxytetracycline-Gentamycin-Nitrofurantoin-Sulphamethoxazole indicated that some isolates were resistant to all of the nine antibiotics tested. Cluster analysis of antibiotic resistance data revealed two major clusters. Sixteen (36.4%), 14 (27.3%), 3 (6.8%), and 2 (4.5%) of the VRE isolates possessed the gel, asa1, hyl, and esp virulence genes respectively while the cylA gene was not detected in the study. Multiple antibiotic-resistant enterococci were also resistant to vancomycin and possessed virulence determinants indicating that they can pose severe public health complications on individuals who consume contaminated water.


VREs Enterococci virulence genes Vancomycin resistance determinants 



The authors would like to appreciate contributions from colleagues of the Department of Microbiology, Faculty of Natural and Agricultural Sciences—North West University.

Funding information

This study was supported by funding provided by the National Research Foundation, HWSETA postgraduate bursary and the North West University Merit Bursary.


  1. Ateba CN, Maribeng MD (2011) Detection of Enterococcus species in groundwater from some rural communities in the Mmabatho area, South Africa: a risk analysis. Afr J Microbiol Res 5:3930–3935CrossRefGoogle Scholar
  2. Ateba CN, Lekoma KP, Kawadza DT (2013) Detection of vanA and vanB genes in vancomycin-resistant enterococci (VRE) from groundwater using multiplex PCR analysis. J Water Health 11:684–691CrossRefGoogle Scholar
  3. Borgen K, Sørum M, Wasteson Y, Kruse H (2001) VanA-type vancomycin-resistant enterococci (VRE) remain prevalent in poultry carcasses 3 years after avoparcin was banned. Int J Food Microbiol 64:89–94CrossRefGoogle Scholar
  4. Cattoir V, Leclercq R (2013) Twenty-five years of shared life with vancomycin-resistant enterococci: is it time to divorce? J Ant Chem 68:731–742Google Scholar
  5. Cetinkaya Y, Falk P, Mayhall CG (2000) Vancomycin-resistant enterococci. Clin Microbiol Rev 13(4):686–707CrossRefGoogle Scholar
  6. Cruickshank R, Duguid J, Marmion B, Swain R (1975) Medical microbiology. 12th edn, Longman Group Limited 2: 34Google Scholar
  7. Da Silva MF, Tiago I, Veríssimo A, Boaventura RA, Nunes OC, Manaia CM (2006) Antibiotic resistance of enterococci and related bacteria in an urban wastewater treatment plant. FEMS Microbiol Ecol 55:322–329CrossRefGoogle Scholar
  8. Depardieu F, Perichon B, Courvalin P (2004) Detection of the van alphabet and identification of enterococci and staphylococci at the species level by multiplex PCR. J Clin Microbiol 42:5857–5860CrossRefGoogle Scholar
  9. Doyle JJ (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15Google Scholar
  10. Fowler VG, Scheld WM, Bayer AS (2015) Eighty two - endocarditis and intravascular infections A2 - Bennett. In: Dolin R, Blaser M (eds) Mandell, Douglas, and Bennett’s principles and practice of infectious diseases, Eighth edn. Content Repository, PhiladelphiaGoogle Scholar
  11. Frieden T, Munsiff S, Williams G, Faur Y, Kreiswirth B, Low D, Willey B, Warren S, Eisner W (1993) Emergence of vancomycin-resistant enterococci in New York City. Lancet 342:76–79CrossRefGoogle Scholar
  12. Goldstein RER, Micallef SA, Gibbs SG, George A, Claye E, Sapkota A, Joseph SW, Sapkota AR (2014) Detection of vancomycin-resistant enterococci (VRE) at four United State wastewater treatment plants that provide effluent for reuse. Sci Total Environ 466:404–411CrossRefGoogle Scholar
  13. Ingram DL, Pearson A, Occhiuti A (1983) Detection of bacterial antigens in body fluids with the Wellcogen Haemophilus influenzae, Streptococcus pneumoniae and Neisseria meningitidis (ACYW135) latex agglutination tests. J Clin Microbiol 18:1119–1121Google Scholar
  14. Jackson CR, Fedorka-Cray PJ, Barrett JB (2004) Use of a genus- and species-specific multiplex PCR for identification of enterococci. J Clin Microbiol 42(8):3558–3565CrossRefGoogle Scholar
  15. Kirby W, Bauer A, Sherris JC, Turck M (1966) Antibiotic susceptibility testing by standardized single disk method. Am J Clin Pathol 45:493CrossRefGoogle Scholar
  16. Klein G (2003) Taxonomy, ecology and antibiotic resistance of enterococci from food and the gastrointestinal tract. Int J Food Microbiol 88(2–3):123–31Google Scholar
  17. Lisboa LF, Miranda BG, Vieira MB, Dulley FL, Fonseca GG, Guimarães T, Levin AS, Shikanai-Yasuda MA, Costa SF (2015) Empiric use of linezolid in febrile hematology and hematopoietic stem cell transplantation patients colonized with vancomycin-resistant Enterococcus species. Int J Inf Dis 33:171–176CrossRefGoogle Scholar
  18. Mohapi IM, Ateba CN (2013) Isolation of vancomycin resistant enterococci isolated from leafy vegetables (lettuce) from North West Province. Life Sci 10(4):1163–1170Google Scholar
  19. Molale L, Bezuidenhout CC (2016) Antibiotic resistance, efflux pump genes and virulence determinants in Enterococcus species from surface water systems. Environ Sci Pollut Res 23:21501–21510CrossRefGoogle Scholar
  20. Ramsey AM, Zilberberg MD (2009) Secular trends of hospitalization with vancomycin-resistant Enterococcus infection in the United States, 2000–2006. ICHE 30:184–186Google Scholar
  21. Ranotkar S, Kumar P, Zutshi S, Prashanth KS, Bezbaruah B, Anand J, Lahkar M (2014) VRE: troublemaker of the 21st centurion. J Glo Ant Res 2:205–212Google Scholar
  22. Roberts MC, Marzluff JM, Delap JH, Turner R (2016) Vancomycin resistant Enterococcus species from crows and their environment in metropolitan Washington State, United States of America: is there a correlation between VRE positive crows and the environment? Vet Microbiol 194:48–54CrossRefGoogle Scholar
  23. Sood S, Malhotra M, Das B, Kapil A (2008) Enterococcal infections and antimicrobial resistance. Ind J Med Res 128:111–121Google Scholar
  24. Takeuchi K, Tomita H, Fujimoto S, Kudo M, Kuwano H, Ike Y (2005) Drug resistance of Enterococcus faecium clinical isolates and the conjugative transfer of gentamicin and erythromycin resistance traits. FEMS Microbiol Lett 243:347–354CrossRefGoogle Scholar
  25. Valenzuela AS, Omar N, Abriouel H, López RL, Ortega E, Cañamero M, Gálvez A (2008) Risk factors in enterococci isolated from foods: Morocco. Food Chem Toxicol 46:2648–2652CrossRefGoogle Scholar
  26. Zirakzadeh A, Patel R (2006) Vancomycin-resistant enterococci: colonization, infection, detection, and treatment. Mayo Clin Proc, Elsevier 81:529–536CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Daniel Pheeha Matlou
    • 1
  • Marie Ebob AgborTabot Bissong
    • 1
  • Christ-Donald Kaptchouang Tchatchouang
    • 1
  • Mohomud Rashid Adem
    • 1
  • Frank Eric Tatsing Foka
    • 1
  • Ajay Kumar
    • 1
  • Collins Njie Ateba
    • 1
    • 2
    Email author
  1. 1.Department of Microbiology, School of Biological Sciences, Faculty of Natural and Agricultural SciencesNorth West UniversityMmabathoSouth Africa
  2. 2.Unit for Environmental Sciences and ManagementNorth-West UniversityPotchefstroomSouth Africa

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