Evaluation of brush cytology (cytospin technique) and cultural results in the diagnosis of keratoconjunctivitis in a goat herd

Abstract

In this study, cytological examination of smears prepared with cytospin and bacteriological isolation from the samples taken with cytobrush from corneal surface of goats with keratoconjunctivitis were evaluated. An outbreak of infectious keratoconjunctivitis, which is particularly affecting kids, was determined in a herd of Saanen goats consisting of 200 animals (120 kids and 80 goats) that located in Bursa province between 2011 and 2012. One hundred and twenty kids were affected severely in this occurrence whereas approximately 10% of the mothers were affected. In clinical examinations, lacrimation and mucopurulent ocular discharge were observed in the kids. In the advanced stages of the illness, the most remarkable lesion was severe corneal edema, which is characterized by opacity and vascularizaton. Although the clinical signs started to appear solely in one eye, both eyes of all the kids were, generally, become affected whereas the mothers had unilateral ocular lesions. The mortality rate in kids reached up to 20% even though they received treatment. All of the infected mothers recovered after treatment. Ocular brush samples were randomly taken from 30 kids for cytological and microbiological examinations. Conjunctival brush samples were examined with cytospin technique to determine whether cellular features can be utilized for cytodiagnosis. The epithelial cells collected with cytobrush were deposited onto poly-L-lysine coated slide glasses by cytocentrifuge. Slides were stained with hematoxylin and eosin (HE). All of the brush samples were cultivated at 37 °C under aerobic conditions for microbiological investigations. The suspicious colonies were selected and subcultured for phenotypical tests. The susceptibility of isolates to the antibiotics was determined via disc diffusion method. In cytological examinations, the commonly encountered cell types in eyes with keratoconjunctivitis were neutrophils with or without bacteria located intracellularly as well as eosinophils, lymphocytes and macrophages. Extracellular bacterial clusters were also observed in some cases. Brush cytology samples taken from eyes with corneal ulcers showed significant reactive cellular changes. The most common bacteria isolated and identified were Staphylococcus aureus, Pseudomonas aeruginosa and Escherichia coli with the rates of 61.11, 50 and 47.22% respectively. All isolated strains had different susceptibility to various antibiotics. Seventy of 120 treated kids recovered within a week but the disease recurred in some of them (n=40). As a result, microbial culture and cytospin preparation of ocular surface samples can contribute for identification of keratoconjunctivitis. This is the first report with regard to the occurrence of keratoconjunctivitis in Saanen goats. Moreover, this study is of importance because cytospin technique can be used to aid in the rapid diagnosis and follow up surveillance of the ocular diseases

Keywords

• Brush cytology
• cytospin technique
• goat
• keratoconjunctivitis

Bir keçi sürüsünde keratokonjunktivitisin teşhisinde fırça sitolojisi (sitospin teknik) ve kültürsonuçlarının değerlendirilmesi

Abstract

Bu çalışmada, keratokonjunktivitisli keçilerin korneal yüzeylerinden fırça ile alınan örneklerin bakteriyolojik izolasyonu ve sitospinle hazırlanmış yaymalarının sitolojik incelenmesi değerlendirildi. 2011-2012 yılları arasında Bursa'da bulunan 200 başlık (120 oğlak ve 80 keçi) bir Saanen keçi sürüsünde, özellikle oğlakları etkileyen, keçilerin bulaşıcı keratokonjunktivitisi saptandı. Bu vakada 120 oğlak şiddetli olarak etkilenirken annelerin %10’ u etkilendi. Oğlaklarda klinik olarak lakrimasyon, mukopurulent oküler akıntı saptandı. Hastalığın ilerlemiş aşamalarında opaklaşma ve vaskülarizasyonla karakterize şiddetli korneal ödem belirlendi. Tüm oğlaklarda klinik bulgular bir gözde görülmeye başlayıp her iki gözü de etkiledi. Analarda ise göz lezyonları tek taraflıydı. Tedavi edilmiş olmalarına rağmen oğlaklarda mortalite oranı %20’ye ulaştı. Tedavi sonrası tüm infekte analarda iyileşme gözlendi. Sitolojik ve mikrobiyolojik incelme için rastgele 30 oğlaktan oküler fırça örnekleri alındı. Fırça ile toplanan epitel hücreleri sitosantrifüj ile poli-L-lisin kaplı lamlara alınmıştır. Lamlar, Hematoxylin-eosin (HE) boyama tekniği ile boyanarak incelenmiştir. Tüm fırça örnekleri, 37 °C' de aerobik şartlar altında mikrobiyolojik incelemeler için kültüre edilmiştir. Şüpheli koloniler seçilmiş ve fenotipik testler için pasajlanmıştır. İzolatlarının antibiyotik duyarlılıkları disk difüzyon yöntemi kullanılarak yapılmıştır. Sitolojik incelemelerde, aktif keratokonjuktivitisli gözlerde yaygın olarak, sitoplazmalarında bakteri bulunan ya da bulunmayan polimorfonüklear nötrofil lökositler ile eozinofil lökositler, lenfosit ve makrofajlar saptandı. Ayrıca bazı örneklerde ekstrasellüler bakteri kümeleri görüldü. Kornea ülserli gözlerden alınan fırça örneklerinde önemli reaktif hücresel değişiklikler mevcuttu.Alınan örneklerin mikrobiyolojik incelemesi sonucunda sırasıyla %61,11, %50 ve %47,22 oranında Staphylococcus aureus, Pseudomonas aeruginosa ve Escherichia coli izole ve identifiye edildi. İzole edilen suşlar çeşitli antibiyotiklere karşı farklı duyarlılıklar gösterdi. Tedavi edilen 70 oğlağın bir hafta içinde iyileştiği ancak bu hayvanların bazılarında (n=40) nüks olduğu görüldü. Sonuç olarak, oküler yüzey örneklerinin mikrobiyal kültürü ve sitosantrifüj ile hazırlanan preparatların sitolojik yönden değerlendirilmesi keratokonjunktivitisin tanısına katkı sağlayabilir. Bu çalışma, Saanen keçilerinde keratokonjunktivitis olgusunu sitospin tekniği ile saptayan ilk çalışmadır. Ayrıca, sitospin tekniğinin oküler yüzey hastalıklarının teşhisi ve takibinde kullanılabilirliğini göstermesi yönünden de bu çalışma önem taşımaktadır

Keywords

• Fırça sitolojisi
• keçi
• keratokonjunktivitis
• sitospin teknik

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