SERAP SAVAŞAN, ŞÜKRÜ KIRKAN, GÖKSEL ERBAŞ, UĞUR PARIN, ALPER ÇİFTCİ

Sığır İzolatı Stafilokokların Virülens Faktörlerinin Belirlenmesi

Stafilokokal mastitisler sığırlar için en önemli ekonomik problemlerden birisidir. Bu çalışmada mastitise neden olan stafilokok türlerinin sahip olduğu virülens faktörlerinin belirlenmesi ve patojenik ile apatojenik türler arasında bu faktörlerin ilişkisinin araştırılması amaçlanmıştır. Çalışma kapsamında 37 S. aureus, 13 S. hyicus, 9 S. simulans, 8 S. chromogenes, 5 S. lentus, 5 S. epidermidis, 2 S. haemolyticus, 2 S. hominis, 1 S. auricularis, 1 S. warneri ve 1 S. sciuri izole ve identifiye edilmiştir. Bu izolatların %41.6'sı koagülaz pozitif olarak belirlenmiştir. Koagülaz pozitif izolatların protein A, DNase, TNase, kapsül, hemoliz, stafilokinaz, slaym (agar'da), biyofilm (mikrodilüsyon) ve hemaglütinasyon özellikleri sırasıyla %71.4, %48.6, %11.4, %40, %97.1, %40, %28.6, % 37.1 ve %, 17.1 olarak tespit edilmiştir. Koagülaz negatif izolatlarda ise bu oranlar sırasıyla %10.2, %12.2, %2, %8.2, %82, %32.7, %12.2, %12.2 ve %10.2 bulunmuştur. Koagülaz pozitif ve negatif izolatlar arasında metisilin direnç oranı %2.9 ve %16.3 olarak saptanmıştır. Sonuç olarak, stafilokok taşıyıcısı hayvanların insanlar için muhtemel infeksiyon kaynağı olabileceği sonucuna varılmıştır.

Determination of Virulence Factors of Staphylococci Isolated from Bovine Mastitis

Staphylococcal mastitis is one of the major economic problems of cattle. The aim of this study was to identify Staphylococcus species that cause mastitis, to examine the virulence factors of these species and to determine the relation between these factors with the pathogenic and nonpathogenic species. In the study, 37 S. aureus, 13 S. hyicus, 9 S. simulans, 8 S. chromogenes, 5 S. lentus, 5 S. epidermidis, 2 S. haemolyticus, 2 S. hominis, 1 S. auricularis, 1 S. warneri, 1 S. sciuri were isolated and identified. The 41.6% of strains were determined as coagulase positive. In the coagulase positive strains, the rate of protein A, DNase, TNase, capsul, hemolyse, staphylokinase, slime (in agar), biofilm (microdilution) and hemagglutination characteristics were found 71.4%, 48.6%, 11.4%, 40%, 97.1%, 40%, 28.6%, 37.1% and 17.1%, respectively. In the coagulase negative strains, the rate of these characteristics were found 10.2%, 12.2%, 2%, 8.2%, 82%, 32.7%, 12.2%, 12.2% and 10.2%, respectively. The methicillin resistance rates in the coagulase positive and negative strains were determined as 2.9 and 16.3%. In conclusion, it was of the opinion that animals are potential carriers of staphylococcus strains that are pathogen for human.

PDF

___

1. Contreras GA, Rodríguez JM: Mastitis: Comparative etiology and epidemiology. J Mammary Gland Biol Neoplasia, 16, 339-356, 2011. DOI: 10.1007/s10911-011-9234-0
2. Kaya O, Gülal M: Aydın yöresinde ineklerde klinik mastitise neden olan mikroorganizmaların saptanması ve bunların antibiyotiklere duyarlılıklarının incelenmesi. Pendik Vet Mikrobiyol Derg, 30 (1): 25-29, 1999.
3. Fitzgerald JR, Hartigan PJ, Meaney WJ, Smyth CJ: Molecular population and virulence factor analysis of Staphylococcus aureus from bovine intramammary infection. J Appl Microbiol, 88, 1028-1037, 2000. DOI: 10.1046/j.1365-2672.2000.01071.x
4. Kawano J, Shimizu A, Saitoh Y, Yagi M, Saito T, Okamoto R: Isolation of methicillin resistance coagulase negative staphylococci from chickens. J Clin Microbiol, 34 (9): 2072-2077, 1996.
5. Quinn PJ, Carter ME, Markey BK, Cartey GR: Clinical Veterinary Microbiology, Vol. 2, 118-126, Mosby-Year Book Europa Limited, Lynton House, London, 2013.
6. Duguid JP: Staining methods. In, Mackie and McCartney: Practical Medical Microbiology. 14th ed., 796-799, Churchill Livingstone, 1996.
7. Gülhan T, Boynukara B, Çiftci A, Söğüt MÜ, Fındık A: Determination of biofilm production, genotype and antibiotic resistance profiles of Enterococcus feacium isolates originated from dog, cat and human. Kafkas Univ Vet Fak Derg, 21 (4): 553-561, 2015. DOI: 10.9775/kvfd.2015.12956
8. Cramton SE, Gerke C, Schnell NF, Nichols WW, Gotz F: The intercellular adhesion (ica) locus is present in Staphylococcus aureus and is required for biofilm formation. Infect Immun, 67 (10): 5427-5433, 1999.
9. Rupp ME, Archer GL: Hemagglutination and adherence to plastic by Staphylococcus epidermidis. Infect Immun, 60 (10): 4322-4327, 1992.
10. Ak S, Horoz H, Ilgaz A: Trakya bölgesinde sığır mastitislerinden sorumlu bulaşıcı ve çevresel bakteriyel etkenler ve antibiyotik duyarlılıkları. Istanbul Univ Vet Fak Derg, 26, 353-365, 2000.
11. Güler L, Ok Ü, Gündüz K, Gülcü Y, Hadimli HH: Antimicrobial susceptibility and coagulase gene typing of Staphylococcus aureus ısolated from bovine clinical mastitis cases in Turkey. J Dairy Sci, 88, 3149- 3154, 2005. DOI: 10.3168/jds.S0022-0302(05)72998-2
12. Kırkan S, Göksoy EÖ, Kaya O: Identification and antimicrobial susceptibility of Staphylococcus aureus and coagulase negative staphylococci from bovine mastitis in the Aydın region of Turkey. Turk J Vet Anim Sci, 29, 791-796, 2005.
13. Türkyılmaz S, Tekbıyık S, Oryasin E, Bozdoğan B: Molecular epidemiology and antimicrobial resistance mechanisms of methicillinresistant Staphylococcus aureus ısolated from bovine milk. Zoonoses Public Hlth, 57, 197-203, 2010. DOI: 10.1111/j.1863-2378.2009.01257.x
14. Wann ER, Fehringer AP, Ezepchuk YU, Schlievert PM, Bina P, Reiser RF, Hook MM, Leung DYM: Staphylococcus aureus isolates from patients with Kawasaki disease Express high levels of protein A. Infect Immun, 67 (9): 4737-4743, 1999.
15. Quinn PJ, Carter ME, Markey BK, Cartey GE: Clinical Veterinary Microbiology. Section 2. Bacteriology, 8. Staphylococcus species. 118-126, Mosby-Year Book Europe Limited, Lynton House, London, England, 1994.
16. Langlois BE, Harmon RJ, Akers K, Aaron DK: Comparison of methods for determining DNase and phosphatase activities of staphylococci. J Clin Microbiol, 27, 1127-1129, 1989.
17. Erganiş O, Kuyucuoglu Y, Ok Ü: İnek ve koyun mastitislerine sebep olan koagulaz negatif ve pozitif stafilokokların biyotiplendirilmesi. Veterinarium, 6, 23-27, 1995.
18. Soell M, Diab M, Haan-archipoff G, Beretz A, Herbelin C, Poutrel B, Klein JP: Capsular polysaccharide type 5 and 8 of Staphylococcus aureus bind specifically to human epithelial (KB) cells, endothelial cells and monocytes and induce release of cytokines. Infect Immun, 63 (4): 1380-1386, 1995.
19. Fattom A, Shepherd S, Karakawa W: Capsular polysaccharide serotyping scheme for Staphylococcus epidermidis. J Clin Microbiol, 30 (12): 3270-3273, 1992.
20. Guidry A, Fattom A, Patel A, O'Brien C: Prevalence of capsular serotypes among Staphylococcus aureus isolates from cows with mastitis in the United States. Vet Microbiol, 59, 53-58, 1997. DOI: 10.1016/S0378- 1135(97)00172-7
21. Poutrel B, Sutra L: Type5 and 8 capsular polysaccharides are expressed by Staphylococcus aureus isolates from rabbits, poultry, pigs and horses. J Clin Microbiol, 31 (2): 467-469, 1993.
22. Ammendolia MG, Di Rosa R, Montanaro L, Arciola CR, Baldassarri L: Slime production and expression of the slime associated antigen by staphylococcal clinical isolates. J Clin Microbiol, 37, 3235-3238, 1999.
23. Baselga R, Albizu I, De LaCruz M, DelCacho E, Barberan M, Amorena B: Phase variation of slime production in Staphylococcus aureus: Implications in colonization and virulence. Infect Immun, 61 (11): 4857-4862, 1993.
24. Dinges MM, Orwin PM, Schlievert PM: Exotoxins of Staphylococcus aureus. Clin Microbiol Rev, 13 (1): 16-34, 2000.
25. Males BM, Rogers WA, Parisi JT: Virulence factors of biotypes of Staphylococcus epidermidis from clinical sources. J Clin Microbiol, 1 (3): 256-261, 1975.
26. Rupp ME, Han J, Gatermann S: Hemagglutination by Staphylococcus aureus strains responsible for human bacteremia or bovine mastitis. Med Microbiol Immunol, 184, 33-36, 1995.
27. Kenar B, Bagcigil AF, Kuyucuoglu Y, Kahraman BB, Konak S: Antimicrobial susceptibility profiles and coagulase gene polymorphism of Staphylococcus aureus isolated from bovine subclinical mastitis. Kafkas Univ Vet Fak Derg, 23 (4): 535-540, 2017. DOI: 10.9775/ kvfd.2016.17247
28. Van duijkeren E, Box ATA, Heck MEOC, Wannet WJB, Fluit AC: Methicillin resistant staphylococci isolated from animals. Vet Microbiol, 103, 91-97, 2004. DOI: 10.1016/j.vetmic.2004.07.014
29. Burriel AR: Resistance of coagulase negative staphylococci isolated from sheep to various antimicrobial agents. Res Vet Sci, 63, 189-190, 1997. DOI: 10.1016/S0034-5288(97)90016-3
30. Juhász-Kaszanyitzky É, Jánosi S, Somogyi P, Dán Á, Bloois L G, Duijkeren E, Wagenaar JA: MRSA transmission between cows and humans. Emerg Infect Dis, 13, 630-632, 2007. DOI: 10.3201/eid1304.060833
31. Leonard FC, Markey BK: Methicillin-Resistant Staphylococcus aureus in Animals. Vet J, 175, 27-36, 2008. DOI: 10.1016/j.tvjl.2006.11.008
32. Stefani S, Varoldo PE: Epidemiology of methicillin resistant staphylococci in Europe. Clin Microbiol Infect, 9, 1179-1189, 2003. DOI: 10.1111/j.1469-0691.2003.00698.x