Skip to main content
SpringerLink
Log in

Evaluation of the extent of spreading of virulence factors and antibiotic resistance in Enterococci isolated from fermented and unfermented foods

  • Original Article
  • Published:
Annals of Microbiology Aims and scope Submit manuscript

Abstract

The aim of this work was to evaluate the impact of fermentation processes on the incidence of virulence and antibiotic resistance determinants in Enterococcus strains from food origin (milk, fermented milk, cheese, fresh meat and fermented meat) with regards to potential pathogenicity. A total of 167 enterococcal strains were used in this study. Of those, 2 were reference strains, and 165 were isolated and identified by molecular methods and screened for virulence factors including ace, agg, ccf, cpd, esp, efaA fm and gelE as well as resistance against ten antibiotics. Enterococcal isolates were identified as Enterococcus faecium (54), Enterococcus faecalis (86), Enterococcus casseliflavus (10) and other Enterococcus spp. (15). Only 3 out of 88 fermented food isolates were free from virulence determinants while approximately 16 % of strains isolated from unfermented foods were free from virulence determinants. Unusually, the Ace gene was detected in E. faecium, especially in fermented foods. In contrast, antibiotic resistance in enterococci was not potentiated by the fermentation process. This study has revealed the important role that may be played by fermentation processes in virulence gene incidence and the potential of such processes to disseminate these traits throughout the enterococci food chain.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

Similar content being viewed by others

References

  • Barbosa J, Gibbs P, Teixeira P (2010) Virulence factors among enterococci isolated from traditional fermented meat products produced in the North of Portugal. Food Control 21:651–656

    Article  Google Scholar 

  • Ben Belgacem Z, Abriouel H, Ben Omar N, Lucas R, Martínez-Canamero M, Gálvez A, Manai M (2010) Antimicrobial activity, safety aspects, and some technological properties of bacteriocinogenic Enterococcus faecium from artisanal Tunisian fermented meat. Food Control 21:462–470

    Article  CAS  Google Scholar 

  • Campos CA, Rodríguez Ó, Calo-Mata P, Prado M, Barros-Velázquez J (2006) Preliminary characterization of bacteriocins from Lactococcus lactis, Enterococcus faecium and Enterococcus mundtii strains isolated from turbot (Psetta maxima). Food Res Int 39:356–364

    Article  CAS  Google Scholar 

  • Canzek Mahjhenic A, Rogelj I, Perko B (2005) Enterococci from Tolminc cheese: population structure, antibiotic susceptibility and incidence of virulence determinants. Int J Food Microbiol 102:239–244

    Article  Google Scholar 

  • Cariolato D, Andrighetto C, Lombardi A (2008) Occurrence of virulence factors and antibiotic resistances in Enterococcus faecalis and Enterococcus faecium collected from dairy and human samples in North Italy. Food Control 19:886–892

    Article  CAS  Google Scholar 

  • Carlos A, Semedo-Lemsaddek T, Barreto-Crespo M, Tenreiro R (2010) Transcriptional analysis of virulence-related genes in enterococci from distinct origins. J Appl Microbiol 108:1563–1575

    Article  CAS  PubMed  Google Scholar 

  • Clewell D (1990) Movable genetic elements and antibiotic resistance in enterococci. Eur J Clin Infect Dis 9(2):90–102

    Article  CAS  Google Scholar 

  • CLSI (2008) Performance standards for antimicrobial disk and dilution susceptibility tests for bacteria isolated from animals; approved standard, 3rd Ed. CLSI document M31-A3. Clinical and Laboratory Standards Institute (CLSI), Wayne, PA

  • Cocconcelli PS, Porro D, Galandini S, Senini L (1995) Development of RAPD protocol for typing of strains of lactic acid bacteria and enterococci. Lett Appl Microbiol 21:376–379

    Article  CAS  PubMed  Google Scholar 

  • Cocconcelli PS, Cattivelli D, Gazzola S (2003) Gene transfer of vancomycin and tetracycline resistances among Enterococcus faecalis during cheese and sausage fermentations. Int J Food Microbiol 88:315–323

    Article  CAS  PubMed  Google Scholar 

  • Dupre I, Zanetti S, Schito A, Fadda G, Sechi L (2003) Incidence of virulence determinants in clinical Enterococcus faecium and Enterococcus faecalis isolates collected from Sardinia (Italy). J Med Microbiol 52:491–498

    Article  CAS  PubMed  Google Scholar 

  • Eaton TJ, Gasson MJ (2001) Molecular screening of Enterococcus virulence determinants and potential for genetic exchange between food and medical isolates. Appl Environ Microbiol 67:1628–1635

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Foulquié-Moreno M, Sarantinopoulos P, Tsakalidou E, de Vuyst L (2006) The role and application of enterococci in food and health. Int J Food Microbiol 106:1–24

    Article  PubMed  Google Scholar 

  • Franz C, Holzapfel W, Stiles M (1999) Enterococci at the crossroads of food safety? Int J Food Microbiol 47:1–24

    Article  CAS  PubMed  Google Scholar 

  • Franz C, Muscholl-Silberhorn A, Yousif N, Vancanneyt M, Swings J, Holzapfel W (2001) Incidence of virulence factors and antibiotic resistance among enterococci isolated from food. Appl Environ Microbiol 67:4385–4389

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Franz C, Huch M, Abriouel H, Holzapfel W, Gálvez A (2011) Enterococci as probiotics and their implications in food safety. Int J Food Microbiol 151:125–140

    Article  CAS  PubMed  Google Scholar 

  • Giraffa G (2003) Functionality of enterococci in dairy products. Int J Food Microbiol 88:215–222

    Article  CAS  PubMed  Google Scholar 

  • Gomes BC, Esteves CT, Palazzo ICV, Darini ALC, Felis GE, Sechi LA, Franco BDG, De Martinis ECP (2008) Prevalence and characterization of Enterococcus spp. isolated from Brazilian foods. Food Microbiol 25:668–675

    Article  CAS  PubMed  Google Scholar 

  • Hirt H, Schlievert PM, Dunny GM (2002) In vivo induction of virulence and antibiotic resistance transfer in Enterococcus faecalis mediated by the sex pheromone-sensing system of pCF10. Infect Immun 70:716–723

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Huycke MM, Sahm DF, Gilmore MS (1998) Multiple-drug resistant enterococci: the nature of the problem and an agenda for the future. Emerging Infect Dis 4:239–249

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Klibi N, Jouini A, Borgo F, Said L, Ferrario C, Dziri R, Boudabous A, Torres C, Slama KB (2015) Antibiotic resistance and virulence of faecal enterococci isolated from food-producing animals in Tunisia. Ann Microbiol 65:695–702. doi:10.1007/s13213-014-0908-x

  • Kumar S, Dudley J, Nei M, Tamura K (2008) MEGA: a biologist-centric software for evolutionary analysis of DNA and protein sequences. Brief Bioinform 9:299–306

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Larkin M, Blackshields G, Brown N, Chenna R, McGettigan PA, McWilliam H, Valentin F, Wallace IM, Wilm A, Lopez R, Thompson JD, Gibson TJ, Higgins DG (2007) Clustal W and Clustal X version 2.0. Bioinformatics 23:2947–2948

    Article  CAS  PubMed  Google Scholar 

  • Martín B, Garriga M, Hugas M, Aymerich T (2005) Genetic diversity and safety aspects of enterococci from slightly fermented sausages. J Appl Microbiol 98:1177–1190

    Article  PubMed  Google Scholar 

  • McCabe KM, Zhang YH, Huang BL, Wagar EA, McCabe ER (1999) Bacterial species identification after DNA amplification with a universal primer pair. Mol Genet Metab 66:205–211

    Article  CAS  PubMed  Google Scholar 

  • McGowan-Spicer LL, Fedorka-Cray PJ, Frye JG, Meinersmann RJ (2008) Antimicrobial resistance and virulence of Enterococcus faecalis isolated from retail food. J Food Prot 71:760–769

    CAS  PubMed  Google Scholar 

  • Miranda J, Guarddon A, Mondragon A, Vazquez B, Fente C, Cepeda A, Franco C (2007) Antimicrobial resistance in Enterococcus spp. strains isolated from organic chicken, and turkey meat: a comparative survey. J Food Prot 70:1021–1024

    CAS  PubMed  Google Scholar 

  • Morrison D, Woodford N, Cookson B (1997) Enterococci as emerging pathogens of humans. J Appl Microbiol Symp Suppl 83:89S–99S

    Article  Google Scholar 

  • Ogier J, Serror P (2008) Safety assessment of dairy microorganisms: The Enterococcus genus. Int J Food Microbiol 126:291–301

    Article  CAS  PubMed  Google Scholar 

  • Ozmen S, Celebi A, Acik L, Temiz A (2010) Virulence gene, antibiotic resistance and plasmid profiles of Enterococcus faecalis and Enterococcus faecium from naturally fermented Turkish foods. J Appl Microbiol 109:1084–1092

    Article  Google Scholar 

  • Peters J, Mac K, Wichmann-Schauer H, Klein G, Ellerbroek L (2003) Species distribution and antibiotic resistance patterns of enterococci isolated from food of animal origin in Germany. Int J Food Microbiol 88:311–314

    Article  CAS  PubMed  Google Scholar 

  • Reviriego C, Eaton T, Martín R, Jiménez E, Fernández L, Gasson M, Rodríguez J (2005) Screening of virulence determinants in Enterococcus faecium strains isolated from breast milk. J Hum Lact 21:131–137

    Article  PubMed  Google Scholar 

  • Saitou N, Nei M (1987) The neighbor-joining method: a new method for reconstructing phylogenetic trees. Mol Biol Evol 4:406–425

    CAS  PubMed  Google Scholar 

  • Santos KM, Vieira AD, Rocha CR, Nascimento JC, Lopes AC, Bruno LM, Carvalho JD, Franco BD, Todorov SD (2014) Brazilian artisanal cheeses as a source of beneficial Enterococcus faecium strains: characterization of the bacteriocinogenic potential. Ann Microbiol 64:1463–1471. doi:10.1007/s13213-013-0789-4

    Article  Google Scholar 

  • Semedo-Lemsaddek T, Nobrega C, Ribeiro T, Pedroso N, Sales-Luís T, Lemsaddek A, Tenreiro R, Tavares L, Vilela C, Oliveira M (2013) Virulence traits and antibiotic resistance among enterococci isolated from Eurasian otter (Lutra lutra). Vet Microbiol 163:378–382

    Article  CAS  PubMed  Google Scholar 

  • Shankar V, Baghdayan AS, Huycke MM, Lindahl G, Gilmore MS (1999) Infection derived Enterococcus faecalis strains are enriched in esp, a gene encoding a novel surface protein. Infect Immun 67:193–200

    CAS  PubMed  PubMed Central  Google Scholar 

  • Templer SP, Baumgartner A (2007) Enterococci from Appenzeller and Schabziger raw milk cheeses: Antibiotic resistance, virulence factors and persistence of particular strains in the products. J Food Prot 70:450–455

    CAS  PubMed  Google Scholar 

  • Valenzuela A, Ben Omar N, Abriouel H, López R, Ortega E, Cañamero M, Gálvez A (2008) Risk factors in enterococci isolated from foods in Morocco: determination of antimicrobial resistance and incidence of virulence traits. Food Chem Toxicol 46:2648–2652

    Article  PubMed  Google Scholar 

  • Valenzuela AS, Omar NB, Abriouel H, López RL, Veljovic K, Cañamero MM, Topisirovic MK, Gálvez A (2009) Virulence factors, antibiotic resistance, and bacteriocins in enterococci from artisan foods of animal origin. Food Control 20:381–385

    Article  CAS  Google Scholar 

  • Wirth R (1994) The sex pheromone system of Enterococcus faecalis. More than plasmid collection mechanisms? Eur J Biochem 222:235–246

    Article  CAS  PubMed  Google Scholar 

  • Xu Y, Kong J (2013) Construction and potential application of controlled autolytic systems for Lactobacillus casei in cheese manufacture. J Food Prot 7:1187–1193

    Article  Google Scholar 

Download references

Acknowledgment

Special appreciation is extended to all the lab team for assistance to accomplish this work.

Author information

Authors and Affiliations

  1. Department of Analytical Chemistry, Nutrition and Food Science, School of Veterinary Sciences, University of Santiago de Compostela, Campus Universitario, 27002, Lugo, Spain

    Alexandre Lamas, Marcos Quintela-Baluja, José Manuel Miranda, Alberto Cepeda & Carlos Manuel Franco

  2. Dairy Department, Faculty of Agriculture, Suez Canal University, 41522, Ismailia, Egypt

    Mohamed Abouelnaga & Magdy Osman

Authors
  1. Mohamed Abouelnaga
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Alexandre Lamas
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Marcos Quintela-Baluja
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Magdy Osman
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. José Manuel Miranda
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Alberto Cepeda
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Carlos Manuel Franco
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Carlos Manuel Franco.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Figure S1

Phylogenetic tree based on 16S rRNA gene sequences, showing the relationships between related enterococcal bacterial species isolated from foodstuffs. Estimates of the statistical significance of phylogenies were calculated by performing 1000 neighbour-joining bootstrap replicates. Scale bar 0.01 substitutions per nucleotide position (DOCX 57 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Abouelnaga, M., Lamas, A., Quintela-Baluja, M. et al. Evaluation of the extent of spreading of virulence factors and antibiotic resistance in Enterococci isolated from fermented and unfermented foods. Ann Microbiol 66, 577–585 (2016). https://doi.org/10.1007/s13213-015-1138-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13213-015-1138-6

Keywords

Search

Navigation