SIĞIRLARDA DOĞAL THEILERIA ANNULATA ENFEKSİYONLARINDA MONOSİT NİTRİK OKSİT DÜZEYLERİ

Abstract

Bu çalışma, Kayseri’nin tropikal theileriosis yönünden enzootik stabiliteye sahip Yeşilhisar ve İncesu yörelerinde Theileria annulata (T.annulata) ile doğal enfekte sığırlarda, enfeksiyon ile monosit nitrik oksit (NO) düzeyleri arasındaki ilişkinin belirlenmesi amacıyla yapılmış olup bu konuda Türkiye’de yapılan ilk çalışmadır. Theileriosis sezonunda, theileriosis şüpheli sığırlardan kan örnekleri alınmış ve kene taşıdığı belirlenen sığırlar üzerindeki keneler toplanmıştır. Negatif kontrol amacıyla Erciyes Üniversitesi Çiftliğindeki sağlıklı sığırlardan kan alınmıştır. T.annulata enfeksiyonu mikroskobik bakı (MB), İndirekt Floresan Antikor Testi (IFAT) ve moleküler metotlarla [Polymerase Chain Reaction (PCR), Reverse Line Blotting (RLB)] araştırılmıştır. Toplanan kenelerin türleri teşhis edilmiş ve T.annulata enfeksiyonu yönünden incelenmişlerdir. Kan örneklerinden lökositler izole edilmiş ve NO düzeyinin göstergesi olan nitrit konsantrasyonu kolorimetrik olarak belirlenmiştir. T.annulata prevalansı, mikroskobik bakı ile %53,33, IFAT ile %60,00, RLB ile %50,00 olarak belirlenmiş, belirlenen prevalans değerleri arasında istatistiksel açıdan önemli fark olmadığı (p>0.05) saptanmıştır. Hayvanların %31,6’sında ergin kene bulunmuştur. Toplanan keneler, Hyalomma anatolicum anatolicum, H.a.excavatum, H.marginatum marginatum ve H.detritum detritum olarak teşhis edilmiş ve bunlardan sadece H.a.anatolicum kenelerinde %20 oranında T.annulata enfeksiyonu saptanmıştır. Nitrik oksit düzeyleri değerlendirildiğinde; akut tropikal theileriosis’li ve agoni halindeki hayvanlarda subklinik ve sağlıklı hayvanlara oranla istatistiki önemde yüksek (p<0.001) bulunmuştur. Hematokrit düzeyi (PCV), hastalık saptanamamış, subklinik enfekte, akut hasta, agoni halindeki hayvanlarda ve kontrol grubu sığırlarda değerlendirildiğinde akut tropikal theileriosis’li ve agoni halindeki hayvanlarda subklinik ve sağlıklı hayvanlara oranla düşük bulunmuştur (p<0.001). Akut tropikal theileriosis’li ve agoni halindeki hayvanların PCV seviyeleri arasındaki farklılığın da istatistiksel açıdan önemli olduğu görülmüştür (p<0.001). Çalışma sonunda sığırların doğal T.annulata enfeksiyonlarında monositlerin NO düzeyinin arttığı gösterilmiştir

Keywords

• Monosit, nitrik oksit, sığır, Theileria annulata

Monocyte Nitric Oxide Levels in Natural Theileria annulata Infections in Cattle

Abstract

This study was performed to determine the monocyte nitric oxide (NO) levels in naturally infected cattle with Theileria annulata (T.annulata) in Yeşilhisar and İncesu districts of Kayseri where the disease is enzootic. This is the first field study investigate the NO levels in monocytes of cattle naturally infected with T. annulata in Turkey. Blood samples and ticks were collected from the animals which were suspected of theileriosis. Blood samples were also collected from the cattle on Erciyes University Farm as negative control. Theileria annulata infection was determined by microscopical examination (ME), Indirect Fluorescent Antibody Technique (IFAT) and molecular (PCR, RLB) examinations. Ticks collected from the animals were identified and examined for the T. annulata infection. Leukocytes were isolated by buffy-coat method and nitrite levels as indicator of NO were determined with a colorimetric method using Griess reagent. The prevalence of T. annulata was 53.33% by ME, 60.00% by IFAT and 50.00% by RLB, and no statistically significant difference was found among these methods (p>0.05). Adult ticks were observed in 31.6% of the animals included in the study, and they were identified as Hyalomma anatolicum anatolicum, H. m. marginatum, H. d. detritum, H. a. excavatum. Only 20% of H. a. anatolicum were found to be naturally infected with T. annulata. The NO levels in animals with acutely and severely infected were significantly higher (p<0.001) than the subclinically infected and control animals. When packet cell volume (PCV) levels were determined in uninfected, subclinically infected, acutely infected, severely infected (agony) and control animals, The PCV in acutely and severely infected animals were significantly lower (p<0.001) than the subclinically infected and control animals. Furthermore, with respect to PCV, the difference between acutely and severely infected animals was also significant (p<0.001). The results of this study to ınvestigate that infected macrophages with T. annulata result in the induction of NO production compared to uninfected cells

Keywords

• Cattle, nitric oxide, monocyte, theileria annulata

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