Safety Evaluation of Enterocin B Producer Enterococcus Faecalis Mye58 Strain Isolated From Raw Milk (Turkish with English Abstract)

Yıl 2014, Cilt: 39 Sayı: 5, 275 - 282, 01.10.2014

Merve Tuncer Banu Özden Tuncer Yasin Tuncer

https://doi.org/10.15237/gida.GD14023

Öz

In this study, we investigated the occurrence of enterocin, virulence factor and vancomycin resistance genes in bacteriocin producer Enterococcus faecalis MYE58 isolated from raw milk. We also tested hemolytic activity, gelatinase production and antibiotic resistance of this strain. It was determined that MYE58 strain inhibited Gram-positive bacteria, such as Listeria monocytogenes, Bacillus cereus and Staphylococcus aureus. The presence of enterocin B structural gene (entB) was detected in MYE58 strain. MYE58 strain did not exhibit haemolysis and gelatinase activity. In MYE58 strain, the presence of gelE and espfs genes were detected, but agg, ace, efaAfs, ccf, cob, cpd, cat, cylM, cylB, cylA, vanA and vanB genes were not detected. MYE58 was found resistant to streptomycin and tetracycline. The results of this study showed that enterocin B producer E. faecalis MYE58 strain used as starter culture could pose a risk to consumer health. However, purified or semi-purified enterocin B produced by MYE58 strain may have a potential to use for food preservation against Listeria monocytogenes, Bacillus cereus and Staphylococcus aureus.

Anahtar Kelimeler

Enterococcus faecalis, enterocin B, antibiotic resistance, virulence factor, raw milk

Çiğ Sütten İzole Edilen Enterosin B Üreticisi Enterococcus Faecalis Mye58 Suşunun Güvenlik Değerlendirmesi

Öz

Bu çalışmada, çiğ sütten izole edilen bakteriyosin üreticisi Enterococcus faecalis MYE58’de enterosin, virülens faktör ve vankomisin direnç genlerinin varlığını araştırdık. Ayrıca, bu suşun hemolitik aktivite, jelatinaz üretimi ve antibiyotik dirençlilik özelliklerini de test ettik. MYE58 suşunun Listeria monocytogenes, Bacillus cereus ve Staphylococcus aureus gibi Gram pozitif bakterileri inhibe ettiği belirlenmiştir. MYE58 suşunda enterosin B yapısal geninin (entB) varlığı tespit edilmiştir. MYE58 suşu hemoliz ve jalatinaz aktivitesi göstermemiştir. MYE58 suşunda gelE ve espfs genlerinin varlığı tespit edilmiş fakat agg, ace, efaAfs, ccf, cob, cpd, cat, cylM, cylB, cylA, vanA ve vanB genlerinin varlığı tespit edilmemiştir. MYE58 suşu streptomisin ve tetrasikline dirençli bulunmuştur. Bu çalışmanın sonuçları, enterosin B üreticisi E. faecalis MYE58 suşunun starter kültür olarak kullanılmasının tüketici sağlığı açısından risk teşkil edebileceğini göstermiştir. Ancak, MYE58 suşu tarafından üretilen enterosin B’nin saflaştırılmış veya kısmi saflaştırılmış preparatlarının Listeria monocytogenes, Bacillus cereus'e Staphylococcus aureus’a karşı gıda muhafazasında kullanılabilme potansiyeli vardır.

Anahtar Kelimeler

Enterococcus faecalis, enterosin B, antibiyotik dirençlilik, virülens faktör, çiğ süt

Kaynakça

• 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.
• Tuncer Y. 2009. Some technological properties of phenotypically identified enterococci strains isolated from Turkish tulum cheese. Afr J Biotechnol, 8 (24), 7008-7016.
• Özden Tuncer B, Ay Z, Tuncer Y. 2013. Occurrence of enterocin genes, virulence factors, and antibiotic resistance 3 bacteriocin-producer Enterococcus faecium strains isolated from Turkish tulum cheese. Turk J Biol, 37, 443-449.
• Yogurtcu NN, Tuncer Y. 2013. Antibiotic susceptibility patterns of Enterococcus strains isolated from Turkish tulum cheese. Int J Dairy Technol, 66, 236-242.
• Ogier JC, Seror P. 2008. Safety assessment of dairy microorganisms: the Enterococcus genus. Int J Food Microbiol, 126, 291-301.
• Khan H, Flint S, Yu PL. 2010. Enterocins in food preservation. Int J Food Microbiol, 141, 1-10.
• Franz CMAP, van Belkum MJ, Holzapfel WH, Abriouel H, Gálvez A. 2007. Diversity of enterococcal bacteriocins and their grouping in a new classification scheme. FEMS Microbiol Rev, 31, 293-310.
• Edalatian MR, Najafi MBH, Mortazavi SA, Alegría A, Delgado S, Bassami MZ, Mayo B. 2012. Production of bacteriocins by Enterococcus spp. isolated from traditional, Iranian, raw milk cheeses, and detection of their encoding genes. Eur Food Res Technol, 234, 789-796.
• Morandi S, Brasca M, Andrighetto C, Lombar- di A, Lodi R. 2006. Technological and molecular characterisation of enterococci isolated from North-West Italian dairy products. Int Dairy J, 16, 867-875.
• 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.
• Ino¤lu ZN, Tuncer Y. 2013. Safety assessment of Enterococcus faecium and Enterococcus facalis strains isolated from Turkish tulum cheese. Journal of Food Safety, 33, 369-377.
• Van Belkum MJ, Hayema BJ, Geis A, Kok J, Venema G. 1989. Clonning of two bacteriocin genes from a lactococcal bacteriocin plasmid. Appl Environ Microbiol, 55, 1187-1191
• Ryan MP, Rea MC, Hill C, Ross RP. 1996. An application in Cheddar cheese manufacture for a strain of Lactococcus lactis producing a novel broad spectrum bacteriocin lacticin 3147. Appl Environ Microbiol, 62, 612-619.
• Cancilla MR, Powell IB, Hillier AJ, Davidson BE. 1992. Rapid genomic fingerprinting of Lactococcus lactis strains by arbitrarily primed polymerase chain reaction with 32P and fluorescent labels. Appl Environ Microbiol, 58, 1772-1775.
• Edwards U, Rogall T, Blocker H, Emde M, Bottger EC. 1989. Isolation and direct complete nucleotide determination of entire genes. Characterization of a gene coding for 16S ribosomal RNA. Nucleic Acids Res, 17, 7843-7853.
• Reviriego C, Eaton T, Martín R, Jiménez E, Fernández L, Gasson MJ, Rodríguez JM. 2005. Screening of virulence determinants of Enterococcus faecium strains isolated from breast milk. Journal of Human Lactation, 21, 131-137.
• CLSI M100-S21. 2011. Performance Standards for Antimicrobial Susceptibility Testing 21th Informational Supplement. Clinical and Laboratory Standards Institute, Wayne, PA, USA.
• Dutka-Malen S, Evers S, Courvalin P. 1995. Detection of glycopeptide resistance genotypes and identification to the species level of clinically relevant enterococci by PCR. Journal of Clinical Microbiology, 33, 24-27.
• De Vuyst L, Vandamme EJ (ed). 1994. Bacteriocins of Lactic Acid Bacteria. Microbiology, Genetics and Applications. Chapman and Hall, New York, USA, 539 p.
• Brandão A, Almeida T, Muñoz-Atienza E, Torres C, Igrejas G, Hernández PE, Cintas LM, Poeta P, Herranz C. 2010. Antimicrobial activity and cccurence of bacteriocin structural genes in Enterococcus spp. of human and animal origin isolated in Portugal. Arch Microbiol, 192, 927-936.
• Favaro L, Basaglia M, Casella S, Hue I, Dousset X, de Melo Franco BDG, Todorov SD. 2014. Bacteriocinogenic potential and safety evaluation of non-starter Enterococcus faecium strains isolated from home made white brine cheese. Food Microbiol, 38, 228-239.
• Pangallo D, Harichova J, Karelova E, Drahovská H, Chovanová K, Ferianc P, Turna J, Timko J. 2004. Molecular investigation of enterococci isolated from different environmental sources. Biologia, Bratislava, 56 (6), 829-837.
• 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 Tunusian fermented meat. Food Control, 21, 462-470.
• Koch S, Hufnagel M, Theilacker C, Huebner C. 2004. Enterococcal infections: host response, therapeutic, and prophylactic possibilities. Vaccine, 22, 822-830.
• Valenzuela AS, Ben Omar N, Abriouel H, López RL, Veljovic K, Cañamero MM, Topisirovic MKL, 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.
• Gomes BC, Esteves CT, Palazzo ICV, Darini AL, Felis GE, Sechi LA, Franco BD, De Martinis EC. 2008. Prevalence and characterization of Enterococcus spp. isolated from Brazilian foods. Food Microbiol, 25, 668-675.
• Upadhyaya PMG, Ravikumar KL, UmapathyBL. 2009. Review of virulence factors of Enterococcus: an emerging nosocomial pathogen. Indian Journal of Medial Microbiology, 27, 301-305.
• Franz CMAP, Muscholl-Silberhorn AB, Yousif NMK, Vancanneyt M, Swings J, Holzapfel WH. 2001. Incidence of virulence factors and antibiotic resistance among enterococci isolated from food. Appl Environ Microbiol, 67, 4385-4389.
• Özmen To¤ay S, Çelebi Keskin A, Açık L, Temiz A. 2010. Virulence genes, antibiotic resistance and plasmid profiles of Enterococcus faecalis and Enterococcus faecium from naturally fermented Turkish foods. J Appl Microbiol, 109, 1084-1092.