Identification and antimicrobial susceptibility of Corynebacterium pseudotuberculosis isolated from sheep

Year 2015, Volume: 31 Issue: 2, 116 - 121, 01.06.2015

Aslı Sakmanoğlu Hasan Hüseyin Hadimli Osman Erganiş Yasemin Pınarkara Zafer Sayın Kürşat Kav

Abstract

Aim: The aim of this study was to identification of Corynebacterium pseudotuberculosis (C. pseudotuberculosis) isolated from lymph node of sheep with caseous lypmhadenitis (CLA) by biochemical characteristics and confirmation by Polymerase Chain Reaction (PCR). Also, phospholipase D enzyme activities and antimicrobial susceptibility of isolates were determined. Materials and Methods: A total of 1176 sheep were examined the presence of CLA, and 1176 of lymph node were collected for bacteria isolation. The samples were inoculated onto Blood Agar Base supplemented with 5% defibrinated sheep blood at 37ºC for 48 hours in microaerophilic atmosphere. Morphological and biochemical characteristic of C. pseudotuberculosis suspicious isolates were determined. PCR analysis was performed to confirmation of biochemical identification. Also, phospholipase D enzyme activities with Rhodococcus equi (R. equi) and Staphylococcus aureus (S. aureus) and antimicrobial susceptibility of isolates for 16 antimicrobial were determined by disc diffusion methods.Results: Totally of 72 C. pseudotuberculosis were identified according to biochemical characteristics. All isolates showed PLD enzyme activities antagonists with S. aureus and synergistically with R. equi. All isolates were confirmed by PCR as C. pseudotuberculosis. Isolates were determined to be sensitive to different antibiotics at different rates. The majority of the isolates were susceptible to florfenicol (98%), while ampicillin and lincomycin resistance was determined as 62.5%.Conclusions: Although there are many data related to the failure of the in-vivo treatment of CLA infection by antibiotics, C. pseudotuberculosis was determined as in-vitro sensitive to many antibiotics used in this study. PCR method and primers used in this study was determined very useful for confirmation of C. pseudotuberculosis identified by biochemical characteristic.

Keywords

Corynebacterium pseudotuberculosis, sheep, caseous lypmhadenitis, PCR

Koyunlardan izole edilen Corynebacterium pseudotuberculosis suşlarının identifikasyonu ve antibiyotiklere duyarlılıkları

Abstract

Amaç: Bu çalışmada, Kazeöz Lenfadenetis (KLA)’li koyunların lenf düğümlerinden izole edilen Corynebacterium pseudotuberculosis (C. pseudotuberculosis) izolatlarının biyokimyasal identifikasyonu, Polimeraz Zincir Reaksiyonu (PZR) ile doğrulanması, fosfolipaz D (PLD) enzim aktivitesinin belirlenmesi ve antibiyotiklere duyarlılıklarının tespit edilmesi amaçlandı. Gereç ve Yöntem: Toplam 1176 adet koyun KLA yönünden muayene edildi ve 1176 lenf düğümü bakteri izolasyonu için alındı. Örnekler, kanlı agara ekildi ve 37°C’de 48 saat mikroaerofilik olarak inkübe edildi. Şüpheli izolatların, morfolojik, biyokimyasal özellikleri PZR analizleri, Rhodococcus equi (R. equi) ve Staphylococcus aureus (S. aureus) ile PLD enzim aktiviteleri belirlendi. Ayrıca, C. pseudotuberculosis izolatlarının 16 antibiyotiğe duyarlılıkları disk diffüzyon metodu ile belirlendi. Bulgular: Toplam 72 şüpheli izolat biyokimyasal özelliklerine göre C. pseudotuberculosis olarak identifiye edildi. Tüm izolatlar S. aureus ile antagonist ve R. equi ile sinerjik olarak PLD enzim aktivitesi gösterdi. İzolatların tümü PZR ile C. pseudotuberculosis olarak doğrulandı. C. pseudotuberculosis izolatlarının farklı antibiyotiklere değişik oranlarda duyarlı oldukları, izolatların büyük çoğunluğunun (%98) florfenikole duyarlı, %62.5’inin ampisilin ve linkomisine dirençli olduğu belirlendi. Öneri: Enfeksiyonun antibiyotiklerle in-vivo tedavide başarısızlığı ile ilgili çok sayıda veri bulunmasına rağmen etkenin bu çalışmada kullanılan pek çok antibiyotiğe in-vitro duyarlı olduğu, PZR'ın ve bu çalışmada kullanılan primerlerin, biyokimyasal özelliklerine göre identifiye edilen izolatların doğrulanmasında faydalı olduğu ifade edilebilir.

Keywords

Corynebacterium pseudotuberculosis, koyun, kazeöz lenfadenitis, PZR

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