Isolation of Enterococcus faecium, characterization of its antimicrobial metabolites and viability in probiotic Minas Frescal cheese
- 68 Downloads
The aim of this study was to isolate Enterococcus faecium from raw milk samples, to characterize its antimicrobial metabolites, and to evaluate its viability in a probiotic Minas Frescal cheese. For this, antagonist activity against Listeria monocytogenes, safety aspects and biochemical, genotypic, and probiotic characteristics of the isolates were evaluated. Minas Frescal cheese was manufactured with the isolate that showed the best characteristics in vitro, and its viability in the product was evaluated. It was observed that of the 478 lactic acid bacteria isolates, only isolate E297 presented antagonist activity, genes encoding for enterocin production and absence of virulence factors. Besides that, E297 presented probiotic characteristics in vitro, and maintained its viability (8.09 log CFU mL−1) for 14 days of cold storage, when it was added to cheese. Therefore, isolate E297 can be considered a promising microorganism for the manufacture of probiotic foods, especially Minas Frescal cheese.
KeywordsBacteriocin-like substances Biopreservation Enterocin LAB Probiotic food
This study was financed in part by Coordenação de Aperfeiçoamento Pessoal de Nível Superior—Brasil (CAPES)—Finance Code 001. The authors would like to thank FAPERGS and CNPq (309101/2016-6) for their financial support for the research.
Compliance with ethical standards
The article complies with all points of ethical standards.
- Aarestrup FM, Agerso Y, Gerner-Smidt P, Madsen M, Jensen LB (2000) Comparison of antimicrobial resistance phenotypes and resistance genes in Enterococcus faecalis and Enterococcus faecium from humans in the community, broilers, and pigs in Denmark. Diagn Microbiol Infect Dis 37:127–137CrossRefGoogle Scholar
- Biavasco F, Foglia G, Paoletti C, Zandri G, Magi G, Guaglianone E, Sundsfjord A, Pruzzo A, Donelli G, Facinelli B (2007) VanA-type enterococci from humans, animals, and food: species distribution, population structure, Tn1546 typing and location, and virulence determinants. Appl Environ Microbiol 73:3307–3319CrossRefGoogle Scholar
- CLSI (Clinical and Laboratory Standards Institute) (2017) Performance Standards for Antimicrobial Susceptibility Testing; Twenty-seven Informational Supplement, CLSI Document M100-S25. PA, USAGoogle Scholar
- FAO/WHO (2001) Evaluation of health and nutritional properties of probiotics in food. Córdoba, Argentina, Food and Agriculture Organization of the United Nations and World Health Organization, pp 1–34Google Scholar
- FAO/WHO (2002) Guidelines for the Evaluation of Probiotics in Food. Food and Agriculture Organization of the United Nations. World Health Organization, London, OntarioGoogle Scholar
- Marco I, Vogel C, Morandin G, Beilke L, Valduga N, Zambiazi E, Martinhago J, Schittler L (2018) Salsichas tipo hot dog: perfil microbiológico, isolamento e caracterização de bactérias ácido láticas (BAL) com potencial antagonista. Revista CSBEA 4:149–162Google Scholar
- Ounissi H, Courvalin P (1987) Appendix B. Nucleotide sequences of streptococcal genes. In: Ferretti JJ, Curtis R (eds) Streptococcal genetics. American Society for Microbiology, Washington DC, p 275Google Scholar
- Perez RH, Himeno K, Ishibashi N, Masuda Y, Zendo T, Fujita K, Wilaipun P, Leelawatcharamas V, Nakayama J, Sonomoto K (2012) Monitoring of the multiple bacteriocin production by Enterococcus faecium NKR-5-3 through a developed liquid chromatography and mass spectrometry-based quantification system. J Biosci Bioeng 114:490–496CrossRefGoogle Scholar
- Prichula J, Zvoboda DDA, Pereira RI, Santestevan NA, Medeiros AW, Motta ADS, Azevedo PA, Giordani AR, Franzzon APG (2013) Perfil de suscetibilidade aos antimicrobianos e diversidade das espécies de enterococos isolados de leite cru de búfalas no Sul do Brasil. Revista Brasileira de Ciência Veterinária 20:104–109CrossRefGoogle Scholar
- Ricciardi A, Blaiotta G, Di Cerbo A, Succi M, Aponte M (2014) Behaviour of lactic acid bacteria populations in Pecorino di Carmasciano cheese samples submitted to environmental conditions prevailing in the gastrointestinal tract: evaluation by means of a polyphasic approach. Int J Food Microbiol 179:64–71CrossRefGoogle Scholar
- Vankerckhoven V, Autgaerden TV, Vael C, Lammens C, Chapelle S, Rossi R, Jabes D, Goossens H (2004) Development of a multiplex PCR for the detection of asa1, gelE, cylA, esp, and hyl genes in enterococci and survey for virulence determinants among european hospital isolates of Enterococcus faecium. J Clin Microbiol 42:4473–4479CrossRefGoogle Scholar
- Van de Klundert JAM, Vliegenthart JS (1993) PCR detection of genes for aminoglycoside-modifying enzymes. In: Persing DH, Smith TF, Tenover FC, White TJ (eds) Diagnostic molecular microbiology. American Society for Microbiology, pp 547–552Google Scholar