Molecular typing of methicillin resistant staphylococcus aureus strains isolated from cows and farm workers

Bu çalışma mastitisli sığır sütleri ile bu sığırlar ve çiftlik çalışanlarının nazal sıvaplarından metisilin dirençli Staphylococcus aureus (MRSA) izolasyon ve identifikasyonu, izolatların moleküler özelliklerinin ve Panton–Valentine Leucocidin (PVL) genlerinin varlığının incelenmesi amaçlandı. Elli altı sığırdan 145 mastitisli süt ile 56 sığır ve 34 çiftlik çalışanına ait nazal sıvaplar alındı. MRSA izolatlarının moleküler özellikleri itilmiş alan jel elektroforezisi (PFGE), stafilokokal protein A (spa), çok lokuslu dizi tiplendirme (MLST) ve stafilokokal kromozom kaset mec (SCCmec) tiplendirmesi ortaya konuldu. Tüm suşlar PVL genleri bakımından negatif idi. Toplam 12 MRSA identifikasyonu yapıldı. PFGE analizi sonucunda 12 MRSA izolatında pulsotip A-D olamak üzere dört pulsotip belirlendi. Pulsotip A olan suşların (n=7) sekans tipi (ST) tipi ST 239, spa tipi t030 olarak bulunurken biri hariç hepsinin SCCmec III taşıdığı tespit edildi. Pulsotip B (n=3) ST1294, t459 ve SCCmec Tip III; pulsotip C (n=1) ST1940, t660, SCCmec Tip III ve pulsotip D (n=1) ST737, t542, SCCmec tip II olarak belirlendi. Çalışma sonucunda, Aydın yöresinde sığır süt, insan ve sığır burun sıvaplarından izole edilmiş olan en yaygın spa tipi t030, MLST tipinin ise ST 239 olduğu belirlendi.

Sığırlardan ve sektör çalışanlarından izole edilen metisilin dirençli stafilokokların moleküler tiplendirmesi

The aims of this study were to isolation and identification of methicillin resistant Staphylococcus aureus (MRSA) from mastitic cow milk and nasal swabs of cows and related workers, investigation presence of Panton-Valentine leucocidin (PVL) genes and molecular characteristics of these isolates. One hundred forty five mastitic milk samples were obtained from of 56 cows and nasal swabs were collected from these cows and 34 farm workers. MRSA isolates were recovered by pulsed field gel electrophoresis (PFGE), staphylococcal chromosomal cassette (SCC) mec typing, staphylococcal protein A (spa) typing, multilocus sequence typing (MLST). A total of 12 MRSA were identified. All strains were PVL genes negative. PFGE analysis of all 12 MRSA isolates produced four distinct pulsotypes (PT) designated as PT A-D. The PT A (7 strains) detected sequence type (ST) were ST 239, spa type t030 and SCCmec III except one. The PT B (3 strains) were ST1294, t459 and SCCmec III, the PT C (1 strain) was ST1940, t660, SCCmec III detected, the PT D (1 strain) was ST737, t542, SCCmec II. Consequently, it can be said that t030 and ST239 was the most common spa and MLST types which isolated from cow milk and nasal swabs and nasal swabs from workers of these farms in Aydın region, Turkey.

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1. Weese S: Methicillin resistant Staphylococcus aureus in animals. Ilar J, 51, 233-244, 2010.
2. Skov R, Smyth R, Larsen AR, Bolmstrôm A, Karlsson A, Mills K, Frimodt-Moller N, Kahlmeter G: Phenotypic detection of methicillin resistance in Staphylococcus aureus by disk diffusion testing and etest on Mueller-Hinton agar. J Clin Microbiol, 44, 4395-4399, 2006.
3. Deurenberg RH, Stobberingh EE: The evolution of Staphylococcus aureus. Infect Genet Evol, 8, 747-763, 2008.
4. Oliveira DC, Lencastre H: Multiplex PCR strategy for rapid identification of structural types and variants of the mec element in methicillin-resistant Staphylococcus aureus. Antimicrob Agents Ch, 46, 2155-2161, 2002.
5. Shopsin, B, Gomez M, Montgomery SO, Smith DH, Waddington M, Dodge DE, Bost DA, Riehman M, Naidich S, Kreiswirth BN: Evaluation of protein A gene polymorphic region DNA sequencing for typing of Staphylococcus aureus strains. J Clin Microbiol, 37, 3556-3563, 1999.
6. Enright MC, Day NPJ, Davies CE, Peacock SJ, Spratt BG: Multilocus sequence typing for characterization of methicillin-resistant and methicillin- susceptible clones of Staphylococcus aureus. J Clin Microbiol, 38, 1008-1015, 2000.
7. Esel D, Doganay M, Alp E, Sumerkan B: Prospective evaluation of blood cultures in Turkey university hospital: Epidemiology, microbiology and patient outcome. Clin Microbiol Infect, 9, 1038-1044, 2003.
8. Alp E, Klaassen CHW, Doganay M, Altoparlak U, Aydin K, Engin A, Kuzucu C, Ozakin C, Ozinel MA, Turhan O, Voss A: MRSA genotypes in Turkey: Persistence over 10 years of a single clone of ST239. J Infection, 58, 433-438, 2009.
9. Aygen B, Yoruk A, Yildiz O, Alp E, Kocagoz S, Sumerkan B, Doganay M: Bloodstream infections caused by Staphylococcus aureus in a university hospital in Turkey: Clinical and molecular epidemiology of methicillin resistant Staphylococcus aureus. Clin Microbiol Infect, 10, 309-314, 2004.
10. Ciftci A, Findik A, Onuk EE, Savasan S: Detection of methicillin resistance and slime factor production of Staphylococcus aureus in bovine mastitis. Brazilian J Microbiol, 40, 254-261, 2009.
11. Inegol E, Turkyilmaz S: Determination of SCCmec types in methicilline resistant staphylococci isolated from cows and farm workers. Ankara Univ Vet Fak Derg, 59, 89-93, 2012.
12. Turkyilmaz S, Tekbiyik S, Oryasin E, Bozdogan B: Molecular epidemiology and antimicrobial resistance mechanisms of methicillin- resistant Staphylococcus aureus isolated from bovine milk. Zoonoses Public Hlth, 57, 197-203, 2010.
13. Graveland H, Wagenaar JA, Broekhuizen-Stins MJ, Oosting- Schothorst I, Schoormans AH, van Duijkeren E, Huijsdens X, Mevius D, Heederik D: Methicillin-resistant Staphylococcus aureus (MRSA) in veal calf farmers and veal calves in the Netherlands. American Society for Microbiology (ASM) Conference on Antimicrobial Resistance in Zoonotic Bacteria and Foodborne Pathogens. Copenhagen, Denmark, 62-63, 2008.
14. Weese JS, Avery BP, Gow S, Booker C, Reid-Smith R: Methicillin resistant Staphylococcus aureus (MRSA) surveillance in slaughter-age pigs and feedlot cattle. American Society for Microbiology (ASM)-ESCMID Conference on Methicillin-Resistant Staphylococci in Animals, London, 2009.
15. Quinn PJ, Carter ME, Markey BK, Cartey GR: Clinical Veterinary Microbiology. Section 2: Bacteriology, 8. Staphylococcus species. 118-126, Mosby-Year Book Europe Limited, London, 1994.
16. Clinical and Laboratory Standards Institute: Performance Standards for Antimicrobial Susceptibility Testing. 15th informational supplement. Approved standard MS100-S16. CLSI, Wayne, PA, 2006.
17. Kim CH, Khan M, Morin DE, Hurley WE, Tripathy DN, Jr Kerhli M, Oluoch AO, Kakoma I: Optimization of the PCR detection of Staphylococcus aureus nuc gene in bovine milk. J Dairy Sci, 84, 74-83, 2001.
18. Lina G, Piemont Y, Godail-Gamot F, Bes M, Peter MO, Gauduchon V, Vandenesch F, Etienne J: Involvement of Panton-Valentine leukocidin- producing Staphylococcus aureus in primary skin infections and pneumonia. Clin Infect Dis, 29, 1128-1132, 1999.
19. Murray BE, Singh KV, Heath JD, Sharma BR, Weinstock GM: Comparison of genomic DNAs of different enterococcal isolates using restriction endonucleases with infrequent recognition sites. J Clin Microbiol, 28, 2059-2063, 1990.
20. Barber M Methicillin-resistant staphylococci. J Clin Pathol, 14, 385- 393, 1961.
21. Devriese LA, Van Damme LR, Famaree L: Methicillin (cloxacillin) resistant Staphylococcus aureus strains isolated from bovine mastitis cases. Zentralbl Veterinarmed B 19, 598-605, 1972.
22. Vanderhaeghen W, Cerpentier T, Adriaensen C, Vicca J, Hermans K, Butaye P: Methicillin-resistant Staphylococcus aureus (MRSA) ST398 associated with clinical and subclinical mastitis in Belgian cows. Vet Microbiol, 144, 166-171, 2010.
23. Murakami K, Minamido W: Identification of methicillin-resistant strains of Staphylococci by polymerase chain reaction. J Clin Microbiol, 29, 2240-2244, 1991.
24. Budimir A, Deurenberg RH, Plecko V, Vink C, Kalenic S, Stobberingh EE: Molecular characterization of methicillin-resistant Staphylococcus aureus bloodstream isolates from Croatia. J Antimicrob Chemoth, 57, 331-334, 2006.
25. Blanc DS, Petignat C, Wenger A, Kuhn G, Vallet Y, Fracheboud D, Trachsel S, Reymond A, Troillet N, Siegrist HH, Oeuvray S, Bes M, Etienne J, Bille J, Francioli P, Zanetti G: Changing molecular epidemiology of methicillin-resistant Staphylococcus aureus in a small geographic area over an eight-year period. J Clin Microbiol, 45, 3729-3736, 2007.
26. Chongtrakool P, Ito T, Ma XX, Kondo Y, Trakulsomboon S, Tiensasitorn C, Jamklang M, Chavalit T, Song JH, Hiramatsu K: Staphylococcal cassette chromosome mec (SCCmec) typing of methicillin resistant Staphylococcus aureus strains isolated in 11 Asian countries: a proposal for a new nomenclature for SCCmec elements. Antimicrob Agents Ch, 50, 1001-1012, 2006.
27. Kilic A, Guclu AU, Senses Z, Bedir O, Aydogan H, Basustaoglu AC: Staphylococcal cassette chromosome mec (SCCmec) characterization and Panton-Valentine leukocidin gene occurrence for methicillin-resistant Staphylococcus aureus in Turkey, from 2003 to 2006. Antonn Leeuw Int J G, 94, 607-614, 2008.
28. Laplana LM, Cepero MA, Ruiz J, Zolezzi PC, Calvo MA, Erazo MC, Gómez-Lus R: Molecular typing of Staphylococcus aureus clinical isolates by pulsed-field gel electrophoresis, staphylococcal cassette chromosome mec type determination and dissemination of antibiotic resistance genes. Int J Antimicrob Ag, 30, 505-513, 2007.
29. Chini V, Petinaki E, Meugnier H, Foka A, Bes M, Etienne J, Dimitracopoulos G, Spiliopoulou I: Emergence of a new clone carrying Panton-Valentine leukocidin genes and staphylococcal cassette chromosome mec type V among methicillin-resistant Staphylococcus aureus in Greece. Scand J Infect Dis, 40, 368-372, 2008.
30. Loeffler A, Lloyd DH: Companion animals: A reservoir for methicillin- resistant Staphylococcus aureus in the community? Epidemiology and Infection, 138, 595-605, 2010.
31. Morgan M: Methicillin resistant Staphylococcus aureus in animal: Zoonosis or humanosis. J Antimicrob Chemoth, 10, 1093-1105, 2008.