Hostname: page-component-586b7cd67f-rcrh6 Total loading time: 0 Render date: 2024-11-25T21:17:16.708Z Has data issue: false hasContentIssue false
  • English
  • Français

Virulence gene profiles in Staphylococcus aureus isolated from cows with subclinical mastitis in eastern Poland

Published online by Cambridge University Press:  01 April 2016

Barbara Kot ,
Piotr Szweda ,
Aneta Frankowska-Maciejewska ,
Małgorzata Piechota  and
Katarzyna Wolska
Barbara Kot*
Affiliation:
Department of Microbiology, Institute of Biology, Siedlce University of Natural Sciences and Humanities, 12 Bolesława Prusa Str., 08-110 Siedlce, Poland
Piotr Szweda
Affiliation:
Department of Pharmaceutical Technology and Biochemistry, Faculty of Chemistry, Gdańsk University of Technology, G. Narutowicza 11/12, 80-233 Gdańsk, Poland
Aneta Frankowska-Maciejewska
Affiliation:
Department of Microbiology, Institute of Biology, Siedlce University of Natural Sciences and Humanities, 12 Bolesława Prusa Str., 08-110 Siedlce, Poland
Małgorzata Piechota
Affiliation:
Department of Microbiology, Institute of Biology, Siedlce University of Natural Sciences and Humanities, 12 Bolesława Prusa Str., 08-110 Siedlce, Poland
Katarzyna Wolska
Affiliation:
Department of Microbiology, Institute of Biology, Siedlce University of Natural Sciences and Humanities, 12 Bolesława Prusa Str., 08-110 Siedlce, Poland
*
*For correspondence; e-mail: [email protected]
Get access Rights & Permissions [Opens in a new window]

Abstract

Staphylococcus aureus is arguably the most important pathogen involved in bovine mastitis. The aim of this study was to determine the virulence gene profiles of 124 Staph. aureus isolates from subclinical mastitis in cows in eastern Poland. The presence of 30 virulence genes encoding adhesins, proteases and superantigenic toxins was investigated by PCR. The 17 different combinations of adhesin genes were identified. Occurrence of eno (91·1%) and fib (82·3%) genes was found to be common. The frequency of other adhesion genes fnbA, fnbB, ebps were 14·5, 50, 25%, respectively, and for cna and bbp were 1·6%. The etA and etD genes, encoding exfoliative toxins, were present in genomes of 5·6 and 8·9% isolates, respectively. The splA and sspA, encoding serine protease, were detected in above 90% isolates. The most frequent enterotoxin genes were sei (21%), sem (19·4%), sen (19·4%), seg (18·5%) and seo (13·7%). The tst gene was harboured by 2·4% isolates. The 19 combinations of the superantigenic toxin genes were obtained and found in 35·5% of isolates. Three of them (seg, sei, sem, sen, seo; sec, seg, sei, sem, sen, seo and seg, sei, sem, sen) were the most frequent and found in 16·1% of the isolates. The most common virulotype, present in 17·7% of the isolates, was fib, eno, fnbB, splA, splE, sspA. The results indicate the variation in the presence of virulence genes in Staph. aureus isolates and considerable diversity of isolates that are able to cause mastitis in cows.

Keywords

Bovine mastitisStaphylococcus aureusvirulence genesvirulence profiles
Type
Research Article
Information
Journal of Dairy Research , Volume 83 , Issue 2 , May 2016 , pp. 228 - 235
Copyright
Copyright © Proprietors of Journal of Dairy Research 2016 

Access options

Get access to the full version of this content by using one of the access options below. (Log in options will check for institutional or personal access. Content may require purchase if you do not have access.)

References

Akineden, Ö, Annemüller, C, Hassan, AA, Lämmler, C, Wolter, W & Zschöck, M 2001 Toxin genes and other characteristics of Staphylococcus aureus isolates from milk of cows with mastitis. Clinical and Diagnostic Laboratory Immunology 8 959–964CrossRefGoogle Scholar
Bania, J, Dąbrowska, A, Bystron, J, Korzekwa, K, Chrzanowska, J & Molenda, J 2006 Distribution of newly described enterotoxin-like genes in Staphylococcus aureus from food. International Journal of Food Microbiology 108 36–41Google Scholar
Becker, K, Roth, R & Peters, G 1998 Rapid and specific detection of toxigenic Staphylococcus aureus: use of two multiplex PCR enzyme immunoassays for amplification and hybridization of staphylococcal enterotoxin genes, exfoliative toxin genes, and toxic shock syndrome toxin 1 gene. Journal of Clinical Microbiology 36 2548–2553Google Scholar
Bystroń, J, Bania, J, Lis, E, Molenda, J & Bednarski, M 2009 Characterisation of Staphylococcus aureus strains isolated from cows’ milk. Bulletin of the Veterinary Institute in Pulawy 53 59–63Google Scholar
Carneiro, CR, Postol, E, Nomizo, R, Reis, LF & Brentani, RR 2004 Identification of enolase as a laminin-binding protein on the surface of Staphylococcus aureus. Microbes and Infection 6 604–608Google Scholar
Downer, R, Roche, F, Park, PW, Mecham, RP & Foster, TJ 2002 The Elastin-binding protein of Staphylococcus aureus (EbpS) ss expressed at the cell surface as an integral membrane protein and not as a cell wall-associated protein. Microbes and Infection 277 243–250Google Scholar
Fournier, C, Kuhnert, P, Frey, J, Miserez, R, Kirchhofer, M, Kaufmann, T, Steiner, A & Graber, HU 2008 Bovine Staphylococcus aureus: association of virulence genes, genotypes and clinical outcome. Research in Veterinary Science 85 439–448Google Scholar
Haveri, MM, Roslöf, A & Pyörälä, S 2008 Molecular types and genetic profiles of Staphylococcus aureus strains isolated from bovine intramammary infections and extramammary sites. Journal of Clinical Microbiology 46 3728–3735CrossRefGoogle ScholarPubMed
Herron-Olson, L, Fitzgerald, JR, Musser, JM & Kapur, V 2007 Molecular correlates of host specialization in Staphylococcus aureus. PLoS ONE 2 e1120CrossRefGoogle ScholarPubMed
Jagielski, T, Puacz, E, Lisowski, A, Siedlecki, P, Dudziak, W, Międzobrodzki, J & Krukowski, H 2014 Antimicrobial susceptibility profiling and genotyping of Staphylococcus aureus isolates from bovine mastitis in Poland. Journal of Dairy Science 97 1–7Google Scholar
Jarraud, S, Peyrat, MA, Lim, A, Tristan, A, Bes, M, Mougel, C, Etienne, J, Vandenesch, F, Bonneville, M & Lina, G 2001 egc, a highly prevalent operon of enterotoxin gene, forms a putative nursery of superantigens in Staphylococcus aureus. Journal of Immunology 166 669–677Google Scholar
Karahan, M, Açik, MN & Cetinkaya, B 2009 Investigation of toxin genes by polymerase chain reaction in Staphylococcus aureus strains isolated from bovine mastitis in Turkey. Foodborne Pathogens and Disease 6 1029–1035CrossRefGoogle ScholarPubMed
Kot, B, Piechota, M, Antos-Bielska, M, Zdunek, E, Wolska, KM, Binek, T, Olszewska, J, Guliński, P & Trafny, EA 2012a Antimicrobial resistance and genotypes of staphylococci from bovine milk and the cowshed environment. Polish Journal of Veterinary Sciences 15 741–749Google Scholar
Kot, B, Piechota, M, Wolska, KM, Frankowska, A, Zdunek, E, Binek, T, Kłopotowska, E & Antosiewicz, M 2012b Phenotypic and genotypic antimicrobial resistance of staphylococci from bovine milk. Polish Journal of Veterinary Sciences 15 677–683CrossRefGoogle ScholarPubMed
Le Loir, Y, Baron, F & Gautier, M 2003 Staphylococcus aureus and food poisoning. Genetics and Molecular Research 2 63–76Google Scholar
Malinowski, E, Lassa, H, Kłossowska, A, Markiewicz, H, Kaczmarowski, M & Smulski, S 2006 Relationship between mastitis agents and somatic cell mount in foremilk samples. Bulletin of the Veterinary Institute in Pulawy 50 349–352Google Scholar
Monday, SR & Bohach, GA 1999 Use of multiplex PCR to detect classical and newly described pyrogenic toxin genes in staphylococcal isolates. Journal of Food Protection 37 3411–3414Google ScholarPubMed
Omoe, K, Hu, D-L, Takahashi-Omoe, H, Nakane, A & Shinagawa, K 2003 Identification and characterization of a new staphylococcal enterotoxin-related putative toxin encoded by two kinds of plasmids. Infection and Immunity 71 6088–6094Google Scholar
Ote, I, Taminiau, B, Duprez, JN, Dizier, I & Mainil, JG 2011 Genotypic characterization by polymerase chain reaction of Staphylococcus aureus isolates associated with bovine mastitis. Veterinary Microbiology 153 285–292Google Scholar
Park, JY, Fox, LK, Seo, KS, McGuire, MA, Park, YH, Rurangirwa, FR, Sischo, WM & Bohach, GA 2011 Detection of classical and newly described staphylococcal superantigen genes in coagulase-negative staphylococci isolated from bovine intramammary infections. Veterinary Microbiology 147 149–154Google Scholar
Patti, JM, Allen, BL, McGavin, MJ & Hook, M 1994 MSCRAMM-mediated adherence of microorganisms to host tissues. Annual Review of Microbiology 48 585–617Google Scholar
Piccinini, R, Borromeo, V & Zecconi, A 2010 Relationship between Staph. aureus gene pattern and dairy herd mastitis prevalence. Veterinary Microbiology 145 100–105CrossRefGoogle ScholarPubMed
Potempa, J, Dubin, A, Korzus, G & Travis, J 1988 Degradation of elastin by a cysteine proteinase from Staphylococcus aureus. Journal of Biological Chemistry 263 2664–2667Google Scholar
Rall, VL, Vieira, FP, Rall, R, Vieitis, RL, Fernandes, A Jr, Candeias, JM, Cardoso, KF & Araújo, JP Jr 2008 PCR detection of staphylococcal enterotoxin genes in Staphylococcus aureus strains isolated from raw and pasteurized milk. Veterinary Microbiology 132 408–413Google Scholar
Rice, K, Huesca, M, Vaz, D & McGavin, MJ 2001 Variance in fibronectin binding and fnb locus polymorphisms in Staphylococcus aureus: identification of antigenic variation in a fibronectin binding protein adhesin of the epidemic CMRSA-1 strain of methicillin-resistant Staph. aureus. Infection and Immunity 69 3791–3799Google Scholar
Rosec, JP & Gigaud, O 2002 Staphylococcal enterotoxin genes of classical and new types detected by PCR in France. International Journal of Food Microbiology 77 61–70Google Scholar
Silveira-Filho, VM, Luz, IS, Campos, AP, Silva, WM, Barros, MP, Medeiros, ES, Freitas, MF, Mota, RA, Sena, MJ & Leal-Balbino, TC 2014 Antibiotic resistance and molecular analysis of Staphylococcus aureus isolated from cow's milk and dairy products in northeast Brazil. Journal of Food Protection 77 583–591CrossRefGoogle ScholarPubMed
Singh, RS, Kumar, R & Yadav, BR 2011 Distribution of pathogenic factors in Staphylococcus aureus strains isolated from intra-mammary infections in cattle and buffaloes. Indian Journal of Biotechnology 10 410–416Google Scholar
Smyth, DS, Hartigan, PJ, Meaney, WJ, Fitzgerald, JR, Deobald, CF, Bohach, GA & Smyth, CJ 2005 Superantigen genes encoded by the egc cluster and SaPIbov are predominant among Staphylococcus aureus isolates from cows, goats, sheep, rabbits and poultry. Journal of Medical Microbiology 54 401–411Google Scholar
Srinivasan, V, Sawant, AA, Gillespie, BE, Headrick, SJ, Ceasaris, L & Oliver, SP 2006 Prevalence of enterotoxin and toxic shock syndrome toxin genes in Staphylococcus aureus isolated from milk of cows with mastitis. Foodborne Pathogens and Disease 3 274–283CrossRefGoogle ScholarPubMed
Stephan, R, Annemüller, C, Hassan, AA & Lämmler, C 2001 Characterization of enterotoxigenic Staphylococcus aureus strains isolated from bovine mastitis in north-east Switzerland. Veterinary Microbiology 78 373–382Google Scholar
Szweda, P, Schielmann, M, Milewski, S, Frankowska, A & Jakubczak, A 2012 Biofilm production and presence of ica and bap genes in Staphylococcus aureus strains isolated from cows with mastitis in the eastern Poland. Polish Journal of Microbiology 61 65–69CrossRefGoogle ScholarPubMed
Szweda, P, Schielmann, M, Frankowska, A, Kot, B & Zalewska, M 2014 Antibiotic resistance in Staphylococcus aureus strains isolated from cows with mastitis in eastern Poland and analysis of susceptibility of resistant strains to alternative nonantibiotic agents: lysostaphin, nisin and polymyxin B. Journal of Veterinary Medical Science 76 355–362Google Scholar
Tristan, A, Ying, L, Bes, M, Etienne, J, Vandenesch, F & Lina, G 2003 Use of Multiplex PCR to identify Staphylococcus aureus adhesins involved in human hematogenous infections. Journal of Clinical Microbiology 41 4465–4467Google Scholar
Van Leeuwen, WB, Melles, DC, Alaidan, A, Al-Ahdal, M, Boelens, HA, Snijders, SV, Wertheim, H, van Duijkeren, E, Peeters, JK, van der Spek, PJ, Gorkink, R, Simons, G, Verbrugh, HA & van Belkum, A 2005 Host- and tissue-specific pathogenic traits of Staphylococcus aureus. Journal of Bacteriology 187 4584–4591Google Scholar
Wang, SC, Wu, CM, Xia, SC, Qi, YH, Xia, LN & Shen, JZ 2009 Distribution of superantigenic toxin genes in Staphylococcus aureus isolates from milk samples of bovine subclinical mastitis cases in two major diary production regions of China. Veterinary Microbiology 137 276–281Google Scholar
Zecconi, A, Cesaris, L, Liandris, E, Daprà, V & Piccinini, R 2006 Role of several Staphylococcus aureus virulence factors on the inflammatory response in bovine mammary gland. Microbial Pathogenesis 40 177–183Google Scholar
Zschöck, M, Riße, K & Sommerhäuser, J 2004 Occurrence and clonal relatedness of sec/tst-gene positive Staphylococcus aureus isolates of quartermilk samples of cows suffering from mastitis. Letters in Applied Microbiology 38 493–498Google Scholar
Zschöck, M, Kloppert, B, Wolter, W, Hamann, HP & Lämmler, Ch 2005 Pattern of enterotoxin genes seg, seh, sei and sej positive Staphylococcus aureus isolated from bovine mastitis. Veterinary Microbiology 108 243–249Google Scholar