The relationship between virulence factors and vancomycin resistance among Enterococci collected from food and human samples in Southern Turkey
Abstract
The aims of this research were to study the prevalence of potential virulence factors, vancomycin resistance and also to evaluate a possible correlation that can exist between vancomycin resistance and potential virulence factors between 51 Enterococcus spp. isolated from food and 50 Enterococcus faecium strains from human in southern Turkey. Identification of the isolates was determined by Vitek-II system. Antimicrobial susceptibility tests were performed by Vitek-II system and disc diffusion method. The presence of vanA and vanB as well as enterococcal virulence genes of cytolysin (cylA), the aggregation substance (asa1), gelatinase (gelE), enterococal surface protein (esp), hyaluronidase (hyl) were investigated by Polymerase Chain Reaction (PCR) method. Haemolysin production was also studied phenotypic method. Apart from one isolate, none of the food originated enterococci were resistant to vancomycin, and none carried vanA and vanB resistance genes. All clinical isolates were resistant to vancomycin and 84% of them carried vanA; 2%, vanB; and 14%, neither vanA nor vanB genes. Except for the cylA gene, all other virulence genes and vancomycin resistance were higher in human strains, and a positive correlation was observed between multivirulence genes and hemolytic activity. For all strains, a positive correlation existed between the esp gene positivity and vancomycin resistance, while for only E. faecium, esp, hyl gene positivity and vancomycin resistance a positive correlation could be seen. Furthermore, “silent cylA” genes were found in two food and one intestinal strains. Based on our findings, we can suggest that virulence increases in parallel to vancomycin resistance, and food may be a potential source for dissemination of gelE, asa1 and hyl virulence genes. Finally, esp and hyl genes presence should carefully be monitored in food originated enterococci
References
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- Archimbaud C, Shankar N, Forestier C, Baghdayan A, Gilmore MS, Charbonne F (2002): In vitro adhesive properties and virulence factors of Enterococcus faecalis strains. Res Microbiol, 53, 75-80.
- Barbosa J, Gibbs PA, Teixeira P (2010): Virulence factors among enterococci isolated from traditional fermented meat products produced in North of Portugal. Food Control, 21, 651-656.
- Baylan O, Nazik H, Bektore B, Citil BE, Turan D, Ongen B, Ozyurt M, Acıkel CH, Haznedaroglu T (2001): The relationship between antibiotic resistance and virulence factors in urinary enterococcus isolates. Mikrobiyol Bul, 453, 430-445.
- Ben Omar N, Castro A, Lucas R., Abrıuel H, Yousıf NMK, Franz CMAP, Holzapfel WH, Perez-Pulido R, Martinez-Canamero M, Galvez A (2004): Functional and safety aspects of enterococci isolated from different spanish foods. Syst Appl Microbiol, 27, 118-130.
- Biavasco F, Foglia G, Paoletti C, Zandri, G, Magi G, Guaglianone E., Sundsfjord A, Pruzzo C, Donelli G, Facinelli, B (2007): VanA-Type Enterococci from humans, animals, and food: species distribution, population structure, n1546 typing and location, and virulence determinants. Appl Environ Microbiol, 7310, 3307–3319.
- CLSI (2011): Performance standards for antimicrobial susceptibility testing, 21th informationel supplement, M 100-S21. Wayne Pa.
- Creti R, Imperi M, Bertuccini L, Faretti F, Orefici G, Di Rosa R (2004): Survey of virulence determinants among Enterococcus faecalis isolated from different sources. J Med Microbiol, 53, 13–20.
- De Fátima Silva Lopes M, Ribeiro T, Abrantes M, Figueiredo Marques JJ, Tenreiro R, Crespo MTB (2005): Antimicrobial resistance profiles of dairy and clinical isolates and type strains of enterococci. Int J Food Microbiol, 103, 191–198.
- Eaton JT, 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.
- 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.
- Franz CMAP, Huch M, Abrıouel H, Holzapfel W, Galvez A (2011): Enterococci as probiotics and their implications in food safety. Int J Food Microbiol, 151, 128-140.
- Foulquie Moreno MR, 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.
- Liu D (2011). Enterococcus. In Semedo-Lemsaddec, Alves, Tenreiro and Crespo. Moleculer detection of human bacterial pathogens. p:239. IN: Tylor &Francis Group LLC, NW.
- Mannu L, Paba A, Daga E, Comunian R, Zanetti S, Dupre` I (2003): Comparison of the incidence of virulence determinants and antibiotic resistance between Enterococcus faecium strains of dairy, animal and clinical origin. Int J Food Microbiol, 88, 291-304.
- Martin 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.
- McGowan-Spicer LL, Fedorka-Cray PJ, Frye JG, Meinersmann RJ, Barrett JB, Jackson CR (2008): Antimicrobial resistance and virulence of Enterococcus faecalis isolated from retail food. J Food Prot, 71, 760-769.
- Ozmen TS, 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.
- Psoni L, Kotzamanides C, Andrighetto C, Lombardi A, Tzanetakis N, Litopoulou-Tzanetaki E (2006): Genotypic and phenotypic heterogeneity in Enterococcus isolates from Batzos, a raw goat milk cheese. Int J Food Microbiol, 109, 109-120.
- Salem-Bekhlit MM, Moussa IMI, Muharram MM, Alanazy FK, Hefni HM (2012): Prevalence and antimicrobial resistance pattern of multidrug-resistent enterococci isolated from clinical specimens. Int J Med Microbiol, 301, 44-51.
- 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, 69, 4366-4372.
- Shankar N, Lockatell CV, Baghdayan AS, Drachenberg C, Gilmore MS, Johnson DE (2001): Role of Enterococcus faecalis surface protein esp in the pathogenesis of ascending urinary tract infection. Infect Immun, 69, 4366-4372.
- Semedo T, Almeida Santos M, Martins P, Silva Lopes MF, Figueiredo Marques JJ, Tenreiro R (2003): Comparative study using type strains and clinical and food isolates to examine haemolytic activity and occurrence of the cyl operon in enterococci. J Clin Microbiol, 41, 2569–2576.
- Sussmuth SD, Muscholl-Silberhorn A, Wirth R, Susa M, Marre R, Rodzinski E (2000): Aggregation substance promotes adherence, phagocytosis and intracellular survival of Enterococcus faecalis within human macrophages and suppresses respiratory burst. Infect Immun, 68, 4900-4906.
- Vankerckhoven V, Van Autgaerden T, 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, 4473-4479.
- Vankerckhoven V, Huys G, Vancanneyt M (2008): Genotypic diversity, antimicrobial resistance, and virulence factors of human isolates and probiotic cultures constituting two intraspecific groups of Enterococcus faecium isolates. Appl Environ Microbiol, 74, 4247-4255.
Türkiye’nin güneyinde gıda ve insan örneklerinden toplanan Enterokok izolatlarının vankomisindirenci ve virülans faktörleri arasındaki ilişki
Abstract
Bu çalışmanın amaçları, Türkiye’nin güneyinde gıda kaynaklı 51 adet Enterococcus spp. ile insan kaynaklı 50 adet Enterococcus faecium izolatlarının potansiyel virülans faktörlerinin, vankomisin direnç özelliklerinin prevalansının belirlenmesi ve vankomisin direnci ile virülans faktörler arasında muhtemel ilişkinin değerlendirilmesidir. İzolatlarının identifikasyonu Vitek-II sistemiyle, antimikrobiyel direnç testleri Vitek-II sistemi ve disk difüzyon metodlarıyla çalışıldı. vanA ve vanB ile sitolizin (cylA), agregasyon faktörü (asa1), jelatinaz (gelE), entrokok yüzey proteini (esp), hiyolüronidaz (hyl), genlerinden oluşan enterokokal virülans genler Polimeraz Zincir Reaksiyonu (PCR) metoduyla, hemolizin üretimi fenotipik yöntemle çalışıldı. Bir izolat dışında, gıda izolatlarının hiçbiri vankomisine dirençli değildi ve hiçbiri vanA ve vanB geni taşımamaktaydı. Klinik izolatların tümü vankomisine dirençliydi ve bu izolatların %84’ü vanA, %2’si vanB genlerini taşımakta; %14’ü vanA ve vanB genlerini taşımamaktaydı. cylA geni dışındaki test edilen diğer virülans genler ve vankomisin direnci, klinik izolatlarda daha yüksekti ve çoklu virülans genler ile vankomisin direnci arasında pozitif bir korelasyon gözlendi. Tüm izolatlar arasında esp gen pozitifliği ile vankomisin direnci, E. faecium türlerinde ise esp, hyl gen pozitiflikleri ile vankomisin direnci arasında pozitif bir korelasyon olduğu görüldü. Ayrıca, iki gıda ve bir intestinal enterokok suşunda ‘’sessiz cylA geni’’ bulundu. Bulgularımıza göre, virülans faktörler vankomisin direncine paralel olarak artabilir ve gıda kaynaklı enterokoklar gelE, asa1, hyl virülans genlerinin yayılmasında potansiyel kaynak olabilir. Sonuç olarak, gıda kaynaklı enterokoklarda esp ve hyl gen varlığı dikkatle izlenmelidir
References
- Abriouel H, Omar NB, Molinos AC, Lopez RL, Grande MJ, Martinez-Viedma P, Ortega E, Canamero A (2008): Comperative analysis of genetic diversity and incidance of virulance factors and antibiotic resistance among enterococcal populations from raw fruit and vagetable foods, water and soil, and clinical samples. Int J Food Microbiol, 123, 38-49.
- Archimbaud C, Shankar N, Forestier C, Baghdayan A, Gilmore MS, Charbonne F (2002): In vitro adhesive properties and virulence factors of Enterococcus faecalis strains. Res Microbiol, 53, 75-80.
- Barbosa J, Gibbs PA, Teixeira P (2010): Virulence factors among enterococci isolated from traditional fermented meat products produced in North of Portugal. Food Control, 21, 651-656.
- Baylan O, Nazik H, Bektore B, Citil BE, Turan D, Ongen B, Ozyurt M, Acıkel CH, Haznedaroglu T (2001): The relationship between antibiotic resistance and virulence factors in urinary enterococcus isolates. Mikrobiyol Bul, 453, 430-445.
- Ben Omar N, Castro A, Lucas R., Abrıuel H, Yousıf NMK, Franz CMAP, Holzapfel WH, Perez-Pulido R, Martinez-Canamero M, Galvez A (2004): Functional and safety aspects of enterococci isolated from different spanish foods. Syst Appl Microbiol, 27, 118-130.
- Biavasco F, Foglia G, Paoletti C, Zandri, G, Magi G, Guaglianone E., Sundsfjord A, Pruzzo C, Donelli G, Facinelli, B (2007): VanA-Type Enterococci from humans, animals, and food: species distribution, population structure, n1546 typing and location, and virulence determinants. Appl Environ Microbiol, 7310, 3307–3319.
- CLSI (2011): Performance standards for antimicrobial susceptibility testing, 21th informationel supplement, M 100-S21. Wayne Pa.
- Creti R, Imperi M, Bertuccini L, Faretti F, Orefici G, Di Rosa R (2004): Survey of virulence determinants among Enterococcus faecalis isolated from different sources. J Med Microbiol, 53, 13–20.
- De Fátima Silva Lopes M, Ribeiro T, Abrantes M, Figueiredo Marques JJ, Tenreiro R, Crespo MTB (2005): Antimicrobial resistance profiles of dairy and clinical isolates and type strains of enterococci. Int J Food Microbiol, 103, 191–198.
- Eaton JT, 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.
- 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.
- Franz CMAP, Huch M, Abrıouel H, Holzapfel W, Galvez A (2011): Enterococci as probiotics and their implications in food safety. Int J Food Microbiol, 151, 128-140.
- Foulquie Moreno MR, 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.
- Liu D (2011). Enterococcus. In Semedo-Lemsaddec, Alves, Tenreiro and Crespo. Moleculer detection of human bacterial pathogens. p:239. IN: Tylor &Francis Group LLC, NW.
- Mannu L, Paba A, Daga E, Comunian R, Zanetti S, Dupre` I (2003): Comparison of the incidence of virulence determinants and antibiotic resistance between Enterococcus faecium strains of dairy, animal and clinical origin. Int J Food Microbiol, 88, 291-304.
- Martin 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.
- McGowan-Spicer LL, Fedorka-Cray PJ, Frye JG, Meinersmann RJ, Barrett JB, Jackson CR (2008): Antimicrobial resistance and virulence of Enterococcus faecalis isolated from retail food. J Food Prot, 71, 760-769.
- Ozmen TS, 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.
- Psoni L, Kotzamanides C, Andrighetto C, Lombardi A, Tzanetakis N, Litopoulou-Tzanetaki E (2006): Genotypic and phenotypic heterogeneity in Enterococcus isolates from Batzos, a raw goat milk cheese. Int J Food Microbiol, 109, 109-120.
- Salem-Bekhlit MM, Moussa IMI, Muharram MM, Alanazy FK, Hefni HM (2012): Prevalence and antimicrobial resistance pattern of multidrug-resistent enterococci isolated from clinical specimens. Int J Med Microbiol, 301, 44-51.
- 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, 69, 4366-4372.
- Shankar N, Lockatell CV, Baghdayan AS, Drachenberg C, Gilmore MS, Johnson DE (2001): Role of Enterococcus faecalis surface protein esp in the pathogenesis of ascending urinary tract infection. Infect Immun, 69, 4366-4372.
- Semedo T, Almeida Santos M, Martins P, Silva Lopes MF, Figueiredo Marques JJ, Tenreiro R (2003): Comparative study using type strains and clinical and food isolates to examine haemolytic activity and occurrence of the cyl operon in enterococci. J Clin Microbiol, 41, 2569–2576.
- Sussmuth SD, Muscholl-Silberhorn A, Wirth R, Susa M, Marre R, Rodzinski E (2000): Aggregation substance promotes adherence, phagocytosis and intracellular survival of Enterococcus faecalis within human macrophages and suppresses respiratory burst. Infect Immun, 68, 4900-4906.
- Vankerckhoven V, Van Autgaerden T, 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, 4473-4479.
- Vankerckhoven V, Huys G, Vancanneyt M (2008): Genotypic diversity, antimicrobial resistance, and virulence factors of human isolates and probiotic cultures constituting two intraspecific groups of Enterococcus faecium isolates. Appl Environ Microbiol, 74, 4247-4255.
Details
| Primary Language | English |
|---|---|
| Subjects | Veterinary Surgery |
| Other ID | JA73DJ72ZG |
| Journal Section | Research Article |
| Authors | |
| Publication Date | June 1, 2014 |
| Published in Issue | Year 2014 |