Özkan ASLANTAŞ, SÜHEYLA TÜRKYILMAZ, Mehmet Ali YILMAZ, EBRU ŞEBNEM YILMAZ

Prevalence of methicillin-resistant staphylococci in dogs

Bu çalışmanın amacı, köpeklerin nazal mukozalarında metisilin dirençli stafilokokların (MRS) varlığınının ve tür dağılımının belirlenmesidir. Bu amaçla, Hatay’da özel veteriner kliniklerine getirilen 162 köpekten nazal svablar alındı. Metisilin direncinin belirlenmesinde 2 μg/ml oksasillin içeren mannitollü tuzlu agar kullanıldı. Bakteriyel identifikasyon 16S rRNA dizi analizi ile gerçekleştirildi. Stafilokokal kromozomal kaset tiplendirmesi (SCCmec) için multipleks polimeraz zincir reaksiyonu (mPZR) yapıldı. Antimikrobiyal duyarlılıkları disk diffuzyon yöntemi ile ve antimikrobiyal direnç genleri PZR ile incelendi. Köpeklerin %15.42’ünden (25/162) mecA geni taşıyan MRS izole edildi. Yirmibeş MRS izolatı S. epidermidis (n=12), S. lentus (n=6), S. hominis (n=4), S. warneri (n=1), S. arlettae (n=1) ve S. haemolyticus (n=1) olarak identifiye edildi. S. arlettae’da mecA geni ilk kez belirlendi. Metisilin dirençli S. aureus (MRSA) ve S. pseudintermedius (MRSP) izole edilmedi. SCCmec tip I, II, III ve IV sırasıyla 1, 10, 9 and 5 MRS izolatında belirlendi. Sonuçlar, MRSA dahil MRS suşlarının ortaya çıkışını belirlemek için sürekli surveyansın gerekli olduğunu işaret etmektedir.

Anahtar Kelimeler:

hastalık yaygınlığı, köpek, ilaç direnci, epidemiyoloji, genler, direnç mekanizmaları, ribozomal RNA, Staphylococcus aureus, Staphylococcus epidermidis, bakteriyel hastalıklar, kromozomlar

Köpeklerde metisilin dirençli stafilokokların prevalansı

The aim of the study was to investigate the occurence and species distribution of methicillin resistant staphylococci (MRS) in the nasal cavity of dogs. Nasal swabs were collected from 162 dogs entering private veterinary clinics in Hatay. Methicillin resistance was detected onto mannitol salt agar containing 2 μg/ml oxacillin and confirmed by mecA Polymerase Chain Reaction (PCR). Bacterial identification was done using 16S rRNA sequencing. Staphylococcal cassette chromosome mec (SCCmec) typing of these isolates were determined by multiplex PCR. Antimicrobial susceptibility testing were performed disk diffusion method and antimicrobial resistance genes were determined by PCR. Methicillin-resistant coagulase negative staphylococci (MRCNS) harbouring mecA were isolated from 15.4% (25/162) of dogs. The species identified were S. epidermidis (n=12), S. lentus (n=6), S. hominis (n=4), S. warneri (n=1), S. arlettae (n=1) and S. haemolyticus (n=1). mecA-mediated methicillin resistance in S. arlettae was described for the first time. Methicillin resistant Staphylococcus aureus (MRSA) and methicillin resistant Staphylococcus pseudintermedius (MRSP) were not detected. SCCmec type I, II, III and IV were identified in 1, 10, 9 and 5 MRS isolates, respectively. The results indicate that continuous surveillance is necessary to determine the emergence of MRS including MRSA.

Keywords:

disease prevalence, dogs, drug resistance, epidemiology, genes, resistance mechanisms, ribosomal RNA, Staphylococcus aureus, Staphylococcus epidermidis, bacterial diseases, chromosomes,

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1. Weese JS, Van Duijkeren E: Methicillin-resistant Staphylococcus aureus and Staphylococcus pseudintermedius in veterinary medicine. Vet Microbiol, 140, 418-429, 2010.
2. Piette A, Verschraegen G: Role of coagulase-negative staphylococci in human disease. Vet Microbiol, 134, 45-54, 2009.
3. Pyörälä S, Taponen S: Coagulase-negative staphylococci-emerging mastitis pathogens. Vet Microbiol, 134, 3-8, 2009.
4. Bagcigil FA, Moodley A, Baptiste KE, Jensen VF, Guardabassi L: Occurrence, species distribution, antimicrobial resistance and clonality of methicillin- and erythromycin-resistant staphylococci in the nasal cavity of domestic animals. Vet Microbiol, 121, 307-315, 2007.
5. Vengust M, Anderson ME, Rousseau J, Weese JS: Methicillin-resistant staphylococcal colonization in clinically normal dogs and horses in the community. Lett Appl Microbiol, 43, 602-606, 2006.
6. Bağcıgil AF, İkiz S, Güzel Ö, Parkan Ç, Ilgaz A: Kliniğe getirilen köpek-lerden ve klinik ortamından metisiline dirençli stafilokokların izolasyonu ve mecA geninin PCR ile araştırılması. VII. Ulusal Veteriner Mikrobiyoloji Kongresi (Uluslararası Katılımlı), 26-28 Eylül 2006, Side-Antalya, 2006.
7. Oliveira DC, de Lencastre H: Multiplex PCR strategy for rapid identification of structural types and variants of the mec element in methicillin-resistant Staphylococcus aureus. Antimicrob Chemother, 46, 2155-61, 2002.
8. Kaynarca S, Türkyılmaz S: Methicillin resistance and slime positivity of staphylococci isolated from bovine mastitis. Kafkas Univ Vet Fak Derg, 16 (4): 567-572, 2010.
9. İnegöl E, Türkyılmaz S: Determination of SCCmec types in methicilline resistant staphylococci isolated from cows and farm workers. Ankara Üniv Vet Fak Derg, 59, 89-93, 2012.
10. Suau A, Bonnet R, Sutren M, Godon JJ, Gibson G, Collins MD, Dore´ J: Direct rDNA community analysis reveals a myriad of novel bacterial lineages within the human gut. Appl Environ Microbiol, 65, 4799-4807, 1999.
11. Sghir A, Antonopoulos D, Mackie RI: Design and evaluation of a lacto- bacillus group-specific ribosomal RNA-targeted hybridization probe and its application to the study of intestinal microecology in pigs. Syst Appl Microbiol, 21, 291-296, 1998.
12. Clinical and Laboratory Standards Institute: Performance standards for Disc and Dilution Susceptibility Tests for Bacteria Isolated from Animals; Approved Standard-third Edition (M31-A3), CLSI, Wayne, PA, 2008
13. Toma E, Barriault D: Antimicrobial activity of fusidic acid and disk diffusion susceptibility testing criteria for Gram-positive cocci. J Clin Microbiol, 33, 1712-1715, 1995.
14. Fuchs PC, Jones RN, Barry AL: Interpretive criteria for disk diffusion susceptibility testing of mupirocin, a topical antibiotic. J Clin Microbiol, 28, 608-609, 1990.
15. Mclaws F, Chopra I, O’Neill AJ: High prevalence of resistance to fusidic acid in clinical isolates of Staphylococcus epidermidis. J Antimicrob Chemother, 61, 1040-1043, 2008.
16. Choi SM, Kim SH, Kim HJ, Lee DG, Choi JH, Yoo JH, Kang JH, Shin WS, Kang MW: Multiplex PCR for the detection of genes encoding aminoglycoside modifying enzymes and methicillin resistance among Staphylococcus species, 18, 631-636, 2003.
17. Strommenger B, Kettlitz C, Werner G, Witte W: Multiplex PCR assay for simultaneous detection of nine clinically relevant antibiotic resistance genes in Staphylococcus aureus. J Clin Microbiol, 41, 4089-4094, 2003.
18. Lüthje P, Schwarz S: Antimicrobial resistance of coagulase-negative staphylococci from bovine subclinical mastitis with particular reference to macrolide-lincosamide resistance phenotypes and genotypes. J Antimicrob Chemother, 57, 966-969, 2006.
19. Martineau F, Picard FJ, Lansac N, Ménard C, Roy PH, Ouellette M, Bergeron MG: Correlation between the resistance genotype determined by multiplex PCR assays and the antibiotic susceptibility patterns of Staphylococcus aureus and Staphylococcus epidermidis. Antimicrob Agents Chemother, 44, 231-238, 2000.
20. Jensen LB, Frimodt-Møller N, Aarestrup FM: Presence of erm gene classes in gram-positive bacteria of animal and human origin in Denmark. FEMS Microbiol Lett, 170, 151-158, 1999.
21. Lina G, Quaglia A, Reverdy ME, Leclercq R, Vandenesch F, Etienne J: Distribution of genes encoding resistance to macrolides, lincosamides, and streptogramins among staphylococci. Antimicrob Agents Chemother, 43, 1062-1066, 1999.
22. Nunes EL, dos Santos KR, Mondino PJ, Bastos Mdo C, Giambiagi-de Marval M: Detection of ileS-2 gene encoding mupirocin resistance in methicillin-resistant Staphylococcus aureus by multiplex PCR. Diagn Microbiol Infect Dis. 34, 77-81, 1999.
23. Köksal F, Yasar H, Samasti M: Antibiotic resistance patterns of coagulase- negative staphylococcus strains isolated from blood cultures of septicemic patients in Turkey. Microbiol Res, 164, 404-410, 2009.
24. Taponen S, Pyörälä S: Coagulase-negative staphylococci as cause of bovinemastitis-Not so different from Staphylococcus aureus? Vet Microbiol, 134, 29-36, 2009.
25. van Duijkeren E, Box AT, Heck ME, Wannet WJ, Fluit AC: Methicillin- resistant staphylococci isolated from animals. Vet Microbiol, 103, 91-97, 2004.
26. Martins A, Cunha de ML: Methicillin resistance in Staphylococcus aureus and coagulase negative staphylococci: epidemiological and molecular aspects. Microbiol Immunol, 51, 787-795, 2007.
27. Kılıç A, Güçlü AÜ, Şenses Z, Bedir B, Aydogan H, Başustaoğlu AC: Staphylococcal cassette chromosome mec (SCCmec) characterization and panton-valentine leukocidin gene occurrence for methicillin-resistant Staphylococcus aureus in Turkey, from 2003 to 2006. Antonie van Leeuwenhoek, 94, 607-614, 2008.
28. Schleifer KH, Kilpper-Balz R, Devriese LA: Staphylococcus arlettae sp. nov., S. equorum sp. nov. and S. kloosii sp. nov.: Three new coagulase-negative, novobiocin-resistant species from animals. Syst Appl Microbiol, 5, 501-509, 1984.
29. Biçer AT: Hastane izolatı Staphylococus aureus ve koagülaz negatif staphylococcus suşlarında metisilin direncinin farklı yöntemlerle araştırıl-ması. Uzmanlık Tezi. Çukurova Üniversitesi Tıp Fakültesi Mikrobiyoloji Anabilim Dalı, Adana, 2009.
30. Hanssen AM, Ericson Sollid JU: SCCmec in staphylococci: genes on the move. FEMS Immunol Med Microbiol, 46, 8-20, 2006.