Haluk ÇEPOĞLU, Mehmet Cemal ADIGÜZEL, Arzu Funda BAĞCIGİL, Seyyal AK

Tavuklarda Vankomisin Dirençli Enterokokların ve Vankomisin Direnci ile İlişkili Genlerin Dağılımının Araştırılm

Bu çalışmada, tavuklarda vankomisin dirençli enterokokların saptanması ve bu direnci kodlayan genlerinin belirlenmesi amacıyla 300 tavuk altlığı ve kloakal svab incelenmiştir. Örnekler Enterococcosel buyyon ve agara ekildi, elde edilen 107 şüpheli izolatın PZR ile cins düzeyinde identifikasyonları yapıldı. Enterococcus spp. olarak saptanan izolatlarının (n=98) fenotipik olarak vankomisin dirençleri disk difuzyonve makro dilusyon yöntemleri ile incelendi. Disk difüzyon yöntemi ile 49’ u dirençli, 48’ i orta duyarlı saptanan izolatların vankomisin MIC değerleri incelendiğinde,5’ i dirençli (MİK≥64 μg/ml), 83’ ü orta duyarlı (MİK =8-16 μg/ml) ve 9’ u duyarlı ( MİK≤4 μg/ml) olarak bulundu. Vankomisin MİK sonucuna göre dirençli ve orta duyarlı çıkan izolatların (n=88), teikoplanin MİK değerleri belirlendi; 2’ si dirençli (MİK≥32 μg/ml) ve 86’ sı duyarlı (MİK≤8μg/ml) olarak bulundu. Vankonmisin dirençli ve orta duyarlı olduğu belirlenen izolatlarının, PZR ile vankomisinle ilişkili direnç genleri ve tür düzeyinde identifikasyonları yapıldı. PZR sonucunda izolatların 5’inin vanAgeni taşıyanE. faecium, 29’unun vanC1 geni taşıyan E. gallinarum, 54’ ünün vanC2/3geni taşıyan E.casseliflavus oldukları belirlendi.

Anahtar Kelimeler:

Dışkı, Enterococcus spp., PZR, Tavuk

Investigation of Vancomycin-Resistant Enterococci and The Distribution of The Vancomycin Resistance Associated Genes in Chickens

In this study, chicken litter and cloacal swabs (n=300) were investigated for the detection of vancomycin-resistant enterococci and determination of resistance associated genes. The samples were cultured in Enterococcocel broth and on Enterococcocel agar. One hundred seven presumptive isolates were examined by PCR at the genus level. For the determination of vancomycin resistance of Enterococcus spp. isolates (n=98), disc diffusion and macro dilution methods were performed. According to disc diffusion test, 49 isolates were resistant and 48 were intermediate and according to the macro-dilution tests, 5 of the isolates were found to be resistant (MIC≥64 μg/ml), 83 of them were intermediate (MIC = 8-16 μg/ml) and 9 of them susceptible (MIC≤4 μg/ml). Vancomycin resistant and intermediate isolates were examined for detection of MIC value for teicoplan (n=88); 2 were resistant (MIC≥32 μg/ml) and 86 susceptible (MIC≤8 μg/ml). Finally, vancomycin resistant and intermediate isolates were examined by PCR for the detection of vancomycin resistance associated genes and identification at species level by PCR. As a result of PCR, 5 of the isolates were vanA gene carrying E. faecium, 29 were E. gallinarum with vanC1 gene and 54 were E. casseliflavus with vanC2/3 gene.

Keywords:

Chicken, Enterococcus spp., Faeces, PCR,

PDF

___

Akgül Ö., Gülhan T. ve Güdücüoğlu H., (2016). Tavuk ve martı kökenli enterokok türlerinin antibiyotik dirençliliğinin fenotipik ve genotipik analizi. Ankara Üniversitesi Veteriner Fakültesi Dergisi, 63, 235-244.
Bağcıgil AF., İkiz S., Ak S. ve Özgür N., (2015). Isolation of vancomycin resistant enterococci from animal faeces, detection of antimicrobial resistance profiles and vancomycin resistant genes. Kafkas Üniversitesi Veteriner Fakültesi Dergisi, 21, 87-94.
Bağcıgil AF., Koenhemsi L., Çelik B., Metiner K., Or ME. ve Ak S., (2016). Examination of vancomycin resistant enterococci (VRE) isolated from canine and feline rectal swabs. İstanbul Üniversitesi Veteriner Fakültesi Dergisi, 42, 111-116.
Bonten MJM., Willems R., Weinstein RA., (2001). Vancomycin-resistant enterococci: why are they here, and where do they come from?. The Lancet Infectious Diseases, 1, 314-325.
Bortolaia V., Mander M., Jensen LB., Olsen JE. and Guardabassi L., (2015). Persistence of vancomycin resistance in multiple clones of Enterococcus faecium isolated from Danish broilers 15 years after the ban of avoparcin. Antimicrobial Agents and Chemotherapy, 59, 2926-2929.
Clinical and Laboratory Standards Institute, (2013). Performance Standards for Antimicrobial Susceptibility Testing; Sixteenth Informational Supplement. Clinical and Laboratory Standards Institute, Wayne, USA, M100-S23.
Coombs GW., Kay ID., Steven RA., Pearman JW., Bertolatti D. and Grubb WB., (1999). Should genotyping testing be done on all phenotypically vancomycin-resistant enterococci detected in hospitals?. Journal of Clinical Microbiology, 37, 1229-1230.
Cortes C., De La Fuente R., Contreras A., Sanches A., Corrales JC., Ruiz-Santa-Quiteria JA. and Orden JA., (2006). Occurence and preliminasry study of antimicrobial resistance of enterococci isolated from dairy goats in Spain. International Journal of Food Microbiology, 110, 100-103.
Çetinkaya Y., (2000). Vankomisin dirençli enterokoklar: epidemiyoloji ve kontrol. Flora, 5, 24-33.
Dilik Z. ve İstanbulluoğlu E., (2010). Studies on phenotyping and genotyping characterization of Enterococcus spp. isolated from entansive broiler farms and rural poultry establishments. Bornova Veteriner Kontrol Araştırma Enstitüsü Dergisi, 32, 37-46.
Dombrádi Z., Bihari Z., Illésné Horváth K. and Szabó J., (2010). Comparison of the VITEK 2 system with the E-test for the determination of glycopeptide susceptibility of vanA and vanC positive enterococci. Acta Microbiologica et Immunologica Hungarica, 57, 157-163.
Elsayed S., Hamilton N., Boyd D. and Mulve M., (2001). Improved primer design for multiplex PCR analysis of vancomycin-resistant Enterococcus spp. Journal of Clinical Microbiology, 39, 2367-2368.