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Genetic relationships and virulence factors in Staphylococcus aureus isolated from raw poultry in South Brazil

Abstract

The genetic relationships in Staphylococcus aureus isolated from raw poultry obtained from various Brazilian broiler chicken processors were analyzed using repetitive extragenic palindromic-polymerase chain reaction (rep-PCR). The distribution of accessory gene regulator (agr) groups was determined, and the presence of biofilm-associated genes and phenotypic features, including biofilm formation and proteolytic, lipolytic, and β-hemolytic activities, was assessed. Isolates were grouped into three major clusters based on rep-PCR fingerprints typed with RW3A primer. The agr group I was the most common genotype identified (86.21 %), followed by groups II (10.34 %) and III (3.45 %). All strains were positive for the sasG gene; the next most frequent genes were icaA (93.1 %) and atlA (51.72 %). Twenty-six of the 29 isolates were biofilm producers. In this study, 96.55 %, 72.41 %, and 62.06 % of the isolates displayed lipolytic, β-hemolytic, and proteolytic activity, respectively. In conclusion, the rep-PCR results suggested a clonal relationship among the S. aureus isolated from raw poultry produced by different broiler chicken processors. Our results also showed that most isolates belonged to agr group I. The presence of biofilm-forming S. aureus strains in raw poultry, their ability to harbor biofilm-associated genes, and the spoilage features that they exhibit are indicative of their pathogenic potential, and may represent a serious problem in the food processing industry.

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References

  • Almeida LM, Zilta M, De Almeida PRB, Mendonça CL, Mamizuka LM (2013) Comparative analysis of agr groups and virulence genes among subclinical and clinical mastitis Staphylococcus aureus isolates from sheep flocks of the Northeast of Brazil. Braz J Microbiol 44:493–498

    PubMed Central  Article  PubMed  Google Scholar 

  • Bibalan MH, Shakeri F, Javid N, Ghaemi A, Ghaemi EA (2014) Accessory gene regulator types of Staphylococcus aureus isolated in Gorgan, North of Iran. J Clin Diagn Res 8:07–09

    Google Scholar 

  • Del Vecchio VG, Petroziello JM, Gress MJ, McCleskey FK, Melcher GP, Crouch HK, Lupski JR (1995) Molecular genotyping of methicillin-resistant Staphylococcus aureus via fluorophore-enhanced repetitive-sequence PCR. J Clin Microbiol 33:2141–2144

    PubMed Central  PubMed  Google Scholar 

  • Devita MD, Wadhera RK, Theis ML, Ingham SC (2007) Assessing the potential of Streptococcus pyogenes and Staphylococcus aureus transfer to foods and customers via a survey of hands, hand-contact surfaces and food-contact surfaces at foodservice facilities. J Food Serv 18:76–79

    Article  Google Scholar 

  • Gharsa H, Sallem RB, Slama KB, Gómez-Sanz E, Lozano C, Jouini A, Klibi N, Zarazaga M, Boudabous A, Torres C (2012) High diversity of genetic lineages and virulence genes in nasal Staphylococcus aureus isolates from donkeys destined to food consumption in Tunisia with predominance of the ruminant associated CC133 lineage. BMC Vet Res 8:203

    PubMed Central  Article  PubMed  Google Scholar 

  • Giaouris E, Heir E, Hébraud M, Chorianopoulos N, Langsrud S, Møretrø T, Habimana O, Desvaux M, Renier S, Nychas GJ (2014) Attachment and biofilm formation by foodborne bacteria in meat processing environments: causes, implications, role of bacterial interactions and control by alternative novel methods. Meat Sci 97:298–309

    Article  PubMed  Google Scholar 

  • Gillet Y, Issartel B, Vanhems P, Fournet JC, Lina G, Bes M, Vandenesch F, Piémont Y, Brousse N, Floret D, Etienne J (2002) Association between Staphylococcus aureus strains carrying gene for Panton-Valentine leukocidin and highly lethal necrotising pneumonia in young immunocompetent patients. Lancet 359:753–759

    CAS  Article  PubMed  Google Scholar 

  • Gilot P, Van Leeuwen JW (2004) Comparative analysis of agr locus diversification and overall genetic variability among bovine and human Staphylococcus aureus isolates. Clin Microbiol 42:1265

    CAS  Article  Google Scholar 

  • Goerke C, Esser S, Kummel M, Wolz C (2005) Staphylococcus aureus strain designation by agr and cap polymorphism typing and delineation of agr diversification by sequence analysis. Int J Med Microbiol 295:67–75

    CAS  Article  PubMed  Google Scholar 

  • Greig JD, Ravel A (2009) Analysis of foodborne outbreak data reported internationally for source attribution. Int J Food Microbiol 130:77–87

    CAS  Article  PubMed  Google Scholar 

  • Gundogan N, Ataol O, Gunal S (2012) Determination of some virulence factors in staphylococci isolated from meat and milk products. Arch Lebensmittelhyg 63:182–186

    Google Scholar 

  • Gundogan N, Ataol O, Torlak FO (2013) Determination of some virulence factors in Staphylococcus aureus, Enterococcus faecalis and Enterococcus faecium isolated from meat and milk products. J Food Safety 33:387–393

    Article  Google Scholar 

  • Gutiérrez D, Delgado S, Vázquez-Sánchez D, Martínez B, Cabo ML, Rodríguez A (2012) Incidence of Staphylococcus aureus and analysis of associated bacterial communities on food industry surfaces. Appl Environ Microbiol 78:8547–8554

    PubMed Central  Article  PubMed  Google Scholar 

  • Hanselman BA, Kruth SA, Rousseau J, Weese JS (2009) Coagulase positive staphylococcal colonization of humans and their household pets. Can Vet 50:954–958

    Google Scholar 

  • Jarraud S, Mougel C, Thioulouse J, Lina G, Meugnier H, Forey F, Nesme X, Etienne J, Vandenesch F (2002) Relationships between Staphylococcus aureus genetic background, virulence factors, agr groups (alleles), and human disease. Infect Immun 70:631–641

    PubMed Central  CAS  Article  PubMed  Google Scholar 

  • Jessen B, Lammert L (2003) Biofilm and disinfection in meat processing plants. Int Biodeterior Biodegrad 51:265–279

    CAS  Article  Google Scholar 

  • Khan S, Rasheed F, Zahra R (2014) Genetic polymorphism of agr locus and antibiotic resistance of Staphylococcus aureus at two hospitals in Pakistan. Pak J Med Sci 30:172–176

    PubMed Central  PubMed  Google Scholar 

  • Martins PD, De Almeida TT, Basso AP, De Moura TM, Frazzon J, Tondo EC, Frazzon APG (2013) Coagulase-positive staphylococci isolated from chicken meat: pathogenic potential and vancomycin resistance. Foodborne Pathog Dis 10:771–776

    CAS  Article  PubMed  Google Scholar 

  • Moise-Broder PA, Sakoulas G, Eliopoulos GM, Schentag JJ, Forrest A, Moellering RCJR (2004) Accessory gene regulator group II polymorphism in methicillin-resistant Staphylococcus aureus is predictive of failure of vancomycin therapy. Clin Infect Dis 38:1700–1705

    CAS  Article  PubMed  Google Scholar 

  • Moura TM, Campos FS, D´Azevedo PA, Van Der Sand ST, Franco AC, Frazzon J, Frazzon APG (2012) Prevalence of enterotoxin-encoding genes and antimicrobial resistance in coagulase-negative and coagulase-positive Staphylococcus isolates from black pudding in Southern Brazil. Rev Soc Bras Med Trop 45:579–585

    Article  PubMed  Google Scholar 

  • Njage PMK, Dolci S, Jans C, Wangoh J, Lacroix C, Meile L (2013) Biodiversity and enterotoxigenic potential of staphylococci isolated from raw and spontaneously fermented camel milk. British Microb Research J 3:128–138

    Article  Google Scholar 

  • Novick RP, Schlievert P, Ruzin A (2001) Pathogenicity and resistance islands of staphylococci. Microbes Infect 3:585–594

    CAS  Article  PubMed  Google Scholar 

  • Otto M (2013) Staphylococcal infections: mechanisms of biofilm maturation and detachment as critical determinants of pathogenicity. Annu Rev Med 64:175–188

    CAS  Article  PubMed  Google Scholar 

  • Parkash M, Rajasekar K, Karmegam N (2007) Bacterial population of raw milk and their proteolytic and lipolytic activities. Res J Bas Appl Sci 3:848–851

    Google Scholar 

  • Peacock SJ, Moore CE, Justice A, Kantzanou M, Story L, Mackie K, O’Neill G, Day NP (2002) Virulent combinations of adhesin and toxin genes in natural populations of Staphylococcus aureus. Infect Immun 70:4987–4996

    PubMed Central  CAS  Article  PubMed  Google Scholar 

  • Pereira V, Lopes C, Castro A, Silva J, Gibbs P, Teixeira P (2009) Characterization for enterotoxin production, virulence factors, and antibiotic susceptibility of Staphylococcus aureus isolates from various foods in Portugal. Food Microbiol 26:278–282

    CAS  Article  PubMed  Google Scholar 

  • Pu S, Wang F, Ge B (2011) Characterization of toxin genes and antimicrobial susceptibility of Staphylococcus aureus isolates from Louisiana retail meats. Foodborne Pathog Dis 8:299–306

    CAS  Article  PubMed  Google Scholar 

  • Reinoso EB, El-Sayedb A, Lämmlerb C, Bognia C, Zscho¨ckc M (2008) Genotyping of Staphylococcus aureus isolated from humans, bovine subclinical mastitis and food samples in Argentina. Microbiol Res 163:314–322

    CAS  Article  PubMed  Google Scholar 

  • Reiter KC, Villa B, Da Silva Paim TG, Sambrano GE, De Oliveira CF, D'azevedo PA (2012) Enhancement of antistaphylococcal activities of six antimicrobials against sasG negative methicillin-susceptible Staphylococcus aureus: an in vitro biofilm model. Diagn Microbiol Infect Dis 74:101–105

    CAS  Article  PubMed  Google Scholar 

  • Roche FM, Meehan M, Foster TJ (2003) The Staphylococcus aureus surface protein SasG and its homologues promote bacterial adherence to human desquamated nasal epithelial cells. Microbiology 149:2759–2767

    CAS  Article  PubMed  Google Scholar 

  • Rodrigues LB, Dos Santos LR, Tagliari VZ, Rizzo NN, Trenhago G, De Oliveira AP, Goetz F, Nascimento VP (2010) Quantification of biofilm production on polystyrene by Listeria, Escherichia coli and Staphylococcus aureus isolated from a poultry slaughterhouse. Braz J Microbiol 41:1082–1085

    PubMed Central  Article  PubMed  Google Scholar 

  • Ruaro A, Andrighetto C, Torriani S, Lombardi A (2013) Biodiversity and characterization of indigenous coagulase-negative staphylococci isolated from raw milk and cheese of North Italy. Food Microbiol 34:106–111

    CAS  Article  PubMed  Google Scholar 

  • Sadoyama G, Santos KR, Brilhante AP, Filho PP (2008) Staphylococcus aureus as source of catheter related bloodstream infection evaluated by PFGE and rep-PCR typing in a Brazilian hospital. APMIS 116:953–960

    CAS  Article  PubMed  Google Scholar 

  • Sakoulas G, Eliopoulos GM, Moellering RCJR, Novick RP, Venkataraman L, Wennersten C, Degirolami PC, Schwaber MJ, Gold HS (2003) Staphylococcus aureus accessory gene regulator (agr) group II: is there a relationship to the development of intermediate-level glycopeptide resistance? J Infect Dis 187:929–938

    CAS  Article  PubMed  Google Scholar 

  • Santos KRN, Fonseca LS, Teixeira LM, Gontijo Filho PP (2001) Typing of Staphylococcus aureus from surgical site infections: comparison of pulsed-field gel electrophoresis (PFGE) and PCR technique using repetitive extragenic palindromic (rep) and Tn916-Shine-Dalgarno (TnSD) target sequences. Int J Med Microbiol 291:231–236

    Article  Google Scholar 

  • Spanu V, Spanu C, Virdis S, Cossu F, Scarano C, De Santis EP (2012) Virulence factors and genetic variability of Staphylococcus aureus strains isolated from raw sheep’s milk cheese. Int J Food Microbiol 153:53–57

    Article  PubMed  Google Scholar 

  • Stepanović S, Vuković D, Hola V, Bonaventura G, Djukić S, Ćirković I, Ruzicka F (2007) Quantification of biofilm in microtiter plates: overview of testing conditions and practical recommendations for assessment of biofilm production by staphylococci. APMIS 115:891–899

    Article  PubMed  Google Scholar 

  • Takeuchi S, Kinoshita T, Kaidoh T, Hashizume N (1999) Purification and characterization of protease produced by Staphylococcus aureus isolated from a diseased chicken. Vet Microbiol 67:195–202

    CAS  Article  PubMed  Google Scholar 

  • Tang J, Chen J, Li H, Zeng P, Li J (2013) Characterization of adhesin genes, staphylococcal nuclease, hemolysis, and biofilm formation among Staphylococcus aureus strains isolated from different sources. Foodborne Pathog Dis 10:757–763

    CAS  Article  PubMed  Google Scholar 

  • Vázquez-Sánchez D, Habimana O, Holck A (2013) Impact of food-related environmental factors on the adherence and biofilm formation of natural Staphylococcus aureus isolates. Curr Microbiol 66:110–121

    Article  PubMed  Google Scholar 

  • Weese JS, Reid-Smith R, Rousseau J, Avery B (2010) Methicillin-resistant Staphylococcus aureus (mrsA) contamination of retail pork. Can Vet J 51:749–52

    PubMed Central  PubMed  Google Scholar 

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Acknowledgments

We thank the Conselho Nacional de Desenvolvimento Científico e Tecnológico do Brasil (CNPq) and the Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) of the Brazilian government for the support received.

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Correspondence to Ana Paula Guedes Frazzon.

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Pinto, J.B., Rossatto, F.C.P., Martins, P.D. et al. Genetic relationships and virulence factors in Staphylococcus aureus isolated from raw poultry in South Brazil. Ann Microbiol 65, 1933–1940 (2015). https://doi.org/10.1007/s13213-014-1031-8

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  • DOI: https://doi.org/10.1007/s13213-014-1031-8

Keywords

  • Staphylococcus aureus
  • Raw poultry
  • agr group
  • rep-PCR
  • Virulence factor