Mastitisli İneklerden İzole Edilen Candida albicans İzolatlarında Virülens Faktörlerinin Belirlenmesi

Abstract

Son yıllarda sığır mastitisinin etiyolojisinde mayaların önemi giderek artmaktadır. Bu çalışmada mastitisli ineklerden toplanan süt örneklerinde Candida albicans’ın neden olduğu mastitis prevalansının belirlenmesi, C. albicans’ın fenotipik ve genotipik yöntemlerle identifikasyonu ve virülens faktörlerinin saptanması amaçlandı. Bu amaçla anket sonuçlarına göre mastitis problemi bulunan 20 süt sığırı işletmesinden 178 hayvandan 686 süt örneği toplandı. Bu örneklerden 49 maya izolatı elde edildi. Fenotipik testlerle (germ tüp testi, klamidospor oluşumu, kromojenik besiyerinde ve 45°C’de üreme) 5 izolat Candida albicans olarak identifiye edildi. Fenotipik testlerle C. albicans olduğu saptanan 5 izolat, C. albicans CALB1 genine spesifik primerler kullanılarak yapılan PZR ile doğrulandı. C. albicans izolatlarında adhezyondan (ALS1) ve fosfalipaz (PLB1) üretiminden sorumlu virülens genleri araştırıldı. İzolatların 2 (%40)’sinde ALS1 ve PLB1, 1 (%20)’inde ALS1 ve 1 (%20)’inde PLB1 genleri saptanırken, 1(%20) izolatta her iki genin de bulunmadığı belirlendi. C. albicans izolatlarının tüp adherens yöntemiyle biyofilm oluşturma özellikleri incelendiğinde 3 (%60) izolatın güçlü pozitif, 1 (%20) izolatın zayıf pozitif ve 1 (%20) izolatın ise negatif olduğu belirlendi. Sonuç olarak, bu çalışma ile mastitisli ineklerden izole edilen C. albicans izolatlarında önemli virülens genleri olan ALS1 ve PLB1 genleri ve biyofilm oluşumu belirlendi.

Keywords

• Candida albicans
• Mastitis
• Virülens genleri.

Determination of Virulence Factors in Candida albicans isolated from Cattle with Mastitis

Abstract

In recent years, the importance of yeast in the etiology of bovine mastitis is increasing. The aim of this study was to determine the prevalence of Candida albicans in milk samples collected from cows with mastitis, to identify the phenotypic and genotypic methods and to determine the virulence factors. According to the results of the surveys, 686 milk samples were collected from 178 animals with mastitis problems from 20 dairy cattle farms. Forty nine yeast isolates were obtained from these samples. Five isolates were identified as C. albicans with phenotypic tests (germ tube test, chlamydospor formation, chromogenic medium and reproduction at 45°C). Five isolates that were found to be phenotypically C. albicans were confirmed by PCR using specific primers for the C. albicans CALB1 gene. It was determined that 2 (%40) of the C. albicans isolates had ALS1 and PLB1, 1 (%20) had ALS1 and 1 (%20) had PLB1 genes and no gene were not found in 1 (%20) isolate. When the biofilm formation properties of C. albicans isolates were examined by tube adherence method, it was determined that 3 (%60) isolates were strong, 1 (%20) isolate was weak positive and 1 (%20) isolate was negative. In conclusion, in the C. albicans isolates which were isolated from cows with mastitis had ALS1 and PLB1 genes and biofilm formation.

Keywords

• Candida albicans
• Mastitis
• Virulence genes.

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