RESEARCH ARTICLE

Enterococcus faecalis can be distinguished from Enterococcus faecium via differential susceptibility to antibiotics and growth and fermentation characteristics on mannitol salt agar

  • Maria L. G. Quiloan 1 ,
  • John Vu 2 ,
  • John Carvalho , 3
Expand
  • 1. Biology Program, Biology Department, California State University Dominguez Hills (CSUDH), Carson, CA 90747, USA
  • 2. M. S. Biology Program, Biology Department, California State University Dominguez Hills (CSUDH), Carson, CA 90747, USA
  • 3. Biology Department, California State University Dominguez Hills (CSUDH), Carson, CA 90747, USA

Received date: 08 Sep 2011

Accepted date: 09 Nov 2011

Published date: 01 Apr 2012

Copyright

2014 Higher Education Press and Springer-Verlag Berlin Heidelberg

Abstract

Enterococcus faecalis and Enterococcus faecium are both human intestinal colonizers frequently used in medical bacteriology teaching laboratories in order to train students in bacterial identification. In addition, hospitals within the United States and around the world commonly isolate these bacteria because they are a cause of bacteremia, urinary tract infections, endocarditis, wound infections, and nosocomial infections. Given that enterococci are becoming more of a world health hazard, it is important for laboratories to be able to distinguish these bacteria within hospitalized patients from other bacterial genera. In addition, laboratories must differentiate different species within the Enterococcus genus as well as different strains within each species. Though enterococci are differentiated from other bacterial genera via classical culture and biochemical methods, nucleic acid sequencing is required to differentiate species within the genus—a costly, time consuming, and technically challenging procedure for laboratory technicians that, in itself, does not necessarily lead to speedy identification of bactericidal antibiotics. In this study, we perform antibiogram analysis to show (1) that penicillin can be rapidly employed to distinguish strains and clinical isolates of E. faecalis from E. faecium, (2) E. faecalis is susceptible to ampicillin, and (3) that vancomycin resistance in enterococci shows no sign of abating. Additionally, we show that E. faecalis can grow on mannitol salt agar and ferment mannitol, while E. faecium lacks these phenotypes. These data reveal that we now have rapid, cost effective ways to identify enterococci to the species, and not just genus, level and have significance for patient treatment in hospitals.

Cite this article

Maria L. G. Quiloan , John Vu , John Carvalho . Enterococcus faecalis can be distinguished from Enterococcus faecium via differential susceptibility to antibiotics and growth and fermentation characteristics on mannitol salt agar[J]. Frontiers in Biology, 2012 , 7(2) : 167 -177 . DOI: 10.1007/s11515-012-1183-5

Acknowledgements

We would like to thank Anthony Cipriano for technical assistance and Gaby Gomez for administrative assistance for this project. This work was supported by a California State University Sally Casanova Memorial RSCAAP research grant, a research grant from the Lois Stokes Alliance for Minority Participation (LS-AMP), and a research grant from The Faculty Legacy Fund of the CSUDH Emeritus Faculty Association.
1
Andersson D I (2003). Persistence of antibiotic resistant bacteria. Curr Opin Microbiol, 6(5): 452-456

DOI PMID

2
Archibald L K, Reller L B (2001). Clinical microbiology in developing countries. Emerg Infect Dis, 7(2): 302-305

DOI PMID

3
Blair E B, Emerson J S, Tull A H (1967). A new medium, salt mannitol plasma agar, for the isolation of Staphylococcus aureus. Am J Clin Pathol, 47(1): 30-39

4
Boe L, Gerdes K, Molin S (1987). Effects of genes exerting growth inhibition and plasmid stability on plasmid maintenance. J Bacteriol, 169(10): 4646-4650

PMID

5
Caldwell B A, Ye C, Griffiths R P, Moyer C L, Morita R Y (1989). Plasmid expression and maintanence during long-term starvation-survival of bacteria in well water. J Environ Microbiol, 55: 1860-1864

6
Carvalho J (2011). Importance of clinical microbiologists for U.S. healthcare infrastructure. Clin Lab Sci, 24(3): 136-141

PMID

7
Cetinkaya Y, Falk P, Mayhall C G (2000). Vancomycin-resistant enterococci. Clin Microbiol Rev, 13(4): 686-707

DOI PMID

8
Chan E D, Iseman M D (2008). Multidrug-resistant and extensively drug-resistant tuberculosis: a review. Curr Opin Infect Dis, 21(6): 587-595

DOI PMID

9
Clark N C, Cooksey R C, Hill B C, Swenson J M, Tenover F C (1993). Characterization of glycopeptide-resistant enterococci from U.S. hospitals. Antimicrob Agents Chemother, 37(11): 2311-2317

PMID

10
Courvalin P (2006). Vancomycin resistance in gram-positive cocci. Clin Infect Dis, 42(Suppl 1): S25-S34

DOI PMID

11
Devriese L, Baele M, Butaye P (2006). The Genus Enterococcus. The Procaryotes. New York: Springer, 163-174

12
Domig K J, Mayer H K, Kneifel W (2003). Methods used for the isolation, enumeration, characterisation and identification of Enterococcus spp. 2. Pheno- and genotypic criteria. Int J Food Microbiol, 88(2-3): 165-188

DOI PMID

13
Elzinga G, Raviglione M C, Maher D (2004). Scale up: meeting targets in global tuberculosis control. Lancet, 363(9411): 814-819

DOI PMID

14
Emori T G, Gaynes R P (1993). An overview of nosocomial infections, including the role of the microbiology laboratory. Clin Microbiol Rev, 6(4): 428-442

PMID

15
Farmer P (2005). Pathologies of Power: Health, Human Rights, and the New War on the Poor. Berkley: University of California Press

16
Fisher K, Phillips C (2009). The ecology, epidemiology and virulence of Enterococcus. Microbiology, 155(6): 1749-1757

DOI PMID

17
Flint S J, Enquest L W, Krug R M, Racaniello V R, Skalka A M (2000). Principles of Virology: Molecular Biology, Pathogenesis, and Control, Washington D C: ASM Press

18
Forbes B A, Sahm D F, Weissfeld A S (2007). Laboratory methods and strategies for antimicrobial susceptibility testing (12th Ed), St. Louis: Mosby Elsevier, 188-189

19
Gerdes K, Rasmussen P B, Molin S (1986). Unique type of plasmid maintenance function: postsegregational killing of plasmid-free cells. Proc Natl Acad Sci USA, 83(10): 3116-3120

DOI PMID

20
Godwin D, Slater J H (1979). The influence of the growth environment on the stability of a drug resistance plasmid in Escherichia coli K12. J Gen Microbiol, 111(1): 201-210

PMID

21
Guerrier-Takada C, Salavati R, Altman S (1997). Phenotypic conversion of drug-resistant bacteria to drug sensitivity. Proc Natl Acad Sci USA, 94(16): 8468-8472

DOI PMID

22
Harrison L (2007). StaphylococciTextbook of Diagnostic Microbiology (3rd Ed), St. Louis, MO: Saunders-Elsevier, 377-379

23
Huycke M M, Sahm D F, Gilmore M S (1998). Multiple-drug resistant enterococci: the nature of the problem and an agenda for the future. Emerg Infect Dis, 4(2): 239-249

DOI PMID

24
Jorgensen J H, Turnridge J D (2007). Susceptibility Test Methods: Dilution and Disk Diffusion Methods. Manual of Clinical Microbiology 9th Edition,Washington D C: ASM Press, 1152-1159

25
Kateete D P, Kimani C N, Katabazi F A, Okeng A, Okee M S, Nanteza A, Joloba M L, Najjuka F C (2010). Identification of Staphylococcus aureus: DNase and Mannitol salt agar improve the efficiency of the tube coagulase test. Ann Clin Microbiol Antimicrob, 9(1): 23

DOI PMID

26
Kües U, Stahl U (1989). Replication of plasmids in gram-negative bacteria. Microbiol Rev, 53(4): 491-516

PMID

27
Leboffe M J, Pierce B E (2005). A Photographic Atlas for the Microbiology Laboratory (3rd Ed), Englewood, CO: Morton Publishing Company, 18-19

28
Lehman D C, Mahon C R, Swarna K (2007). Streptococcus, Enterococcus, and other catalase-negative gram-positive cocci. Textbook of Diagnostic Microbiology (3rd Ed), St Louis, MO: Saunders-Elsevier, 382-409

29
Lenski R E, Bouma J E (1987). Effects of segregation and selection on instability of plasmid pACYC184 in Escherichia coli B. J Bacteriol, 169(11): 5314-5316

PMID

30
Lightfoot N F, Scot R J D, Turnball P C B (1990). Antimicrobial susceptibility of Bacillus anthracis. Salisbury Med Bull, 68(Suppl): 95-98

31
Megran D W (1992). Enterococcal endocarditis. Clin Infect Dis, 15(1): 63-71

DOI PMID

32
Modi R I, Adams J (1991). Coevolution in bacterial-plasmid populations. Evolution, 45(3): 656-667

DOI

33
Moellering R C Jr (1992). Emergence of Enterococcus as a significant pathogen. Clin Infect Dis, 14(6): 1173-1178

DOI PMID

34
Murray P R, Rosenthal K S, Pfaller M A (2009). Enterococcus and other gram-positive cocci. Medical Microbiology (6th Ed), Philadelphia: Mosby Elsevie243-246

35
Noble W C, Virani Z, Cree R G (1992). Co-transfer of vancomycin and other resistance genes from Enterococcus faecalis NCTC 12201 to Staphylococcus aureus. FEMS Microbiol Lett, 93(2): 195-198

DOI PMID

36
Pang T, Peeling R W (2007). Diagnostic tests for infectious diseases in the developing world: two sides of the coin. Trans R Soc Trop Med Hyg, 101(9): 856-857

DOI PMID

37
Rhee J I, Ricci J C D, Bode J, Schugerl K (1994). Metabolic enhancement due to plasmid maintenance. Biotechnol Lett, 16: 881-884

DOI

38
Rhode C (1995). Technical information sheet No. 12: plasmid isolation from bacteria: some fast procedures. World J Microbiol Biotechnol, 11(3): 367-369

DOI

39
Rice L B (2006). Antimicrobial resistance in gram-positive bacteria. Am J Med, 119(6 Suppl 1): S11-S19, discussion S62-S70

DOI PMID

40
Seo J H, Bailey J E (1985). Effects of recombinant plasmid content on growth properties and cloned gene product formation in Escherichia coli. Biotechnol Bioeng, 27(12): 1668-1674

DOI PMID

41
Smith M A, Bidochka M J (1998). Bacterial fitness and plasmid loss: the importance of culture conditions and plasmid size. Can J Microbiol, 44(4): 351-355

DOI PMID

42
Stetler H C, Granade T C, Nunez C A, Meza R, Terrell S, Amador L, George J R (1997). Field evaluation of rapid HIV serologic tests for screening and confirming HIV-1 infection in Honduras. AIDS, 11(3): 369-375

DOI PMID

43
Sung J M L, Lindsay J A (2007). Staphylococcus aureus strains that are hypersusceptible to resistance gene transfer from enterococci. Antimicrob Agents Chemother, 51(6): 2189-2191

DOI PMID

44
Teixeira L M, Carvalho M G S, Shewmaker P L, Facklam R R (2011). Manual of Clinical Microbiology. Washington, D C: ASM Press, 350-364

45
Usdin M, Guillerm M, Calmy A (2010). Patient needs and point-of-care requirements for HIV load testing in resource-limited settings. J Infect Dis, 201(s1 Suppl 1): S73-S77

DOI PMID

46
Valenzuela M S, Ikpeazu E V, Siddiqui K A (1996). E. coli growth inhibition by a high copy number derivative of plasmid pBR322. Biochem Biophys Res Commun, 219(3): 876-883

DOI PMID

47
Vu J, Carvalho J (2011). Enterococcus: review of its physiology, pathogenesis, diseases and the challenges it poses for clinical microbiology. Front Biol, 6(5): 357-366

DOI

48
Wilkinson D, Wilkinson N, Lombard C, Martin D, Smith A, Floyd K, Ballard R (1997). On-site HIV testing in resource-poor settings: is one rapid test enough? AIDS, 11(3): 377-381

DOI PMID

49
Woodford N, Johnson A P, Morrison D, Speller D C (1995). Current perspectives on glycopeptide resistance. Clin Microbiol Rev, 8(4): 585-615

PMID

50
Yamahara K M, Layton B A, Santoro A E, Boehm A B (2007). Beach sands along the California coast are diffuse sources of fecal bacteria to coastal waters. Environ Sci Technol, 41(13): 4515-4521

DOI PMID

Outlines

/