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
References
- Abbas B, Razig SA (1979): The etiology of keratoconjunctivitis occurring in goats in the Sudan. Vet Rec, 105, 348-350.
- Atkins KA, Powers CN ( 2002): The cytopathology of infectious diseases. Adv Anat Pathol, 9, 52-64.
- Bankemper KW, Lindley DM, Nusbaum KE, Mysinger RH (1990): Keratoconjunctivitis associated with Neisseria ovis infection in a herd of goats. J Vet Diagn Invest, 2, 76-78.
- Bauer AW, Kirby WMM, Sherris JC, Turck M (1966): Antibiotic susceptibility testing by a standardized single disk method. Am J Clin Pathol, 36, 493-496.
- Bauer GA, Spiess BM, Lutz H (1996): Exfoliative cytology of conjunctiva and cornea in domestic animals: a comparison of four collecting techniques. Vet Comp Ophthalmol, 6, 181-186.
- Belloy L, Janovsky M, Vilei EM, Pilo P, Giacomettim M, Frey J (2003): Molecular epidemiology of Mycoplasma conjunctivae in caprinae: Transmission across species in natural outbreaks. Appl Environ Microbiol, 69, 1913- 1919.
- Bolzan, A, Brunelli AJ, Castro M, Souza M, Souza J, Laus J (2005): Conjunctival impression cytology in dogs. Vet ophthalmol, 8, 401-405.
- Busch TJ, Belton DR (1988): Infectious keratoconjunctivitis in goats. N Z Vet J, 36, 153-155.
- Çakır L, Hızlısoy H, Abay S, Kutsal O (2011): Investigation of ocular surface cytology (brush-impression techniques) and Bacteriological Agents in dogs with Conjunctivitis and Healthy. Cytopathology, 22, 19.
- Dagnall GJ (1994): Use of exfoliative cytology in the diagnosis of ovine keratoconjunctivitis. Vet Rec, 135, 127- 130.
- Dubielzig RR, Ketring KL, McLellan GJ, Albert DM (2010): Veterinary Ocular Pathology A Comparative Review. 1st Edition. Saunders Elsevier, Oxford.
- Finger PT, Papp C, Latkany P, Kurli M, Iacob CE (2006):Anterior chamber paracentesis cytology (cytospin technique) for the diagnosis of intraocular lymphoma. Br J Ophthalmol, 6, 690-692.
- Fujihara T, Takeuchi T, Saito K, Kitajima Y, Kobayashi TK, Tsubato K (1997): Evaluation of Human Conjunctival Epithelium by a Combination of Brush Cytology and Flow Cytometry: An Approach to the Quantitative Technique. Diagn Cytopathol, 17, 456-460.
- Giacometti M, Janovsky M, Belloy L, Frey J (2002): Infectious keratoconjunctivitis of ibex, chamois and other Caprinae. Rev Sci Tech Off Int Épizoot, 21, 335–345.
- Gilger BC (2006): Ocular cytology-your key to immediate ocular diagnosis. 871-872. In: Proceedings of the North American Veterinary Conference, NAVG Gainesville, FL, USA.
- Hillström A, Tvedten H, Maria Källberg M, Hanås S, Lindhe A, Holst BS (2012): Evaluation of cytologic findings in feline conjunctivitis. Vet Clin Path, 41, 283–290.
- Jégou JP, Liotet S (1993): The benefit of conjunctival scraping cytology in the biological diagnosis of conjunctivitis in the dog and cat. EJCAP, 3, 83–97.
- Jones GE (1991): Infectious keratoconjunctivitis. 280– 283. In: Martin WB., Aitken ID., Editors: Diseases of sheep, Blackwell Scientific Publications.
- König CD (1983): Keratoconjunctivitis infectiosa ovis (KIO), 'pink eye' or 'zere oogjes' (a survey). Vet Q, 5, 127- 130.
- Lavach JD, Thrall MA, Benjamin MM, Severin GA (1977). Cytology of normal and inflamed conjunctivas in dogs and cats. J Am Vet Med Assoc, 170, 722–727.
- Levine TS, Rolfe KJ, Crow J, Styles S, Perrett CW, Maclean AB, Reid WMN (2001): The use of cytospin monolayer technique in the cytological diagnosis of vulval and anal disease. Cytopathology, 12, 297–305.
- Maggs DJ (2001): Laboratory investigation of ophthalmic disease. 24-29. In: Petersen-Jones SM, Crispin S (Eds), BSAVA Manual of Small Animal Ophthalmology. Second Edition. British Small Animal Veterinary Association, England.
- Marco I, Mentaberre G, Ballesteros C, Bischof DF, Lavín S, Vilei EM (2009): First Report of Mycoplasma conjunctivae from Wild Caprinae with Infectious Keratoconjunctivitis in the Pyrenees (NE Spain). J Wildlife Dis, 45, 238-241.
- Massa KL, Murphy CJ, Hartmann FA, Miller PE, Korsower CS, Young KM (1999): Usefulness of aerobic microbial culture and cytological evaluation of corneal specimens in the diagnosis of infectious ulcerative keratitis in animals. J Am Vet Med Assoc, 215,1671-1674.
- Mayer D, Nicolet J, Giacometti M, Schmitt M, Wahli T, Meier W (1996): Isolation of Mycoplasma conjunctivae from conjunctival swabs of Alpine ibex (Capra ibex ibex) affected with infectious keratoconjunctivitis. J Vet Med Ser B, 43, 155-161.
- Mushi EZ, Binta MG, Chabo RG, Dintwe K (2007): Conjunctival Flora of Fifty Healthy Goats in Sebele Farm, Gaborone, Botswana. J Anim Vet Adv, 6, 1388-1389.
- Prado MR, Rocha MFG, Brito ÉHS, Girão MD, Monterio AJ, Teixeira MFS, Sidrim JC (2005): Survey of bacterial microorganisms in the conjunctival sac of clinically normal dogs and dogs with ulcerative keratitis in Fortaleza, Ceará, Brazil. Vet Ophthalmol, 8, 33-37.
- Shanholtzer CJ, Schaper PJ, Peterson LR (1982): Concentrated gram stain smears prepared with a cytospin centrifuge. J Clin Microbiol, 16, 1052–1056.
- Sharma S (2012): Diagnosis of infectious diseases of the eye. Eye, 26,177-84.
- Takano Y, Fukagawa K, Dogru M, Asano-Kato N, Tsubota K, and Fujishima H (2004): Inflammatory cells in brush cytology samples correlate with the severity of corneal lesions in atopic keratoconjunctivitis. Br J Ophthalmol, 88, 1504–1505.
- Ter Laak EA, Schreuder BE, Smith-Buys CM (1998): The occurance of Mycoplasma conjunctivae in the Netherlands and its association with infectious kerato- conjunctivitis in sheep and goats. Vet Quart, 10, 73-83.
- Tschopp R, Frey J, Zimmermann L, Giacometti M (2005): Outbreaks of infectious keratoconjunctivitis in alpine chamois and ibex in Switzerland between 2001 and 2003. Vet Rec, 157,13-18.
- Uchida Y (1990): Viral diseases of the outer eye–rapid diagnosis by immunohistochemistry. Nippon Ganka Gakkai Zasshi, 94, 889–902.
- Wakamatsu TH, Okada N, Kojima T, Matsumoto Y, Ibrahim, OMA, Adan ES, Fukagawa K, Katakami C, Tsubota K, Shimazaki J, Fujishima H (2009): Evaluation of conjunctival inflammatory status by confocal scanning laser microscopy and conjunctival brush cytology in patients with atopic keratoconjunctivitis (AKC). Mol Vis, 15, 1611–1619.
- Willis M, Bounous DI, Hirsh S, Kaswan R, Stiles J, Martin C, Rakich P, Roberts W (1997): Conjunctival brush cytology: evaluation of a new cytological collection technique in dogs and cats with a comparison to conjunctival scraping. Vet Comp Ophthalmol, 7, 74-81.
- Yağmur M, Ersöz C, Ersöz TR, Varinli S (1997): Brush technique in ocular surface cytology. Diagn Cytopathol, 17, 88-91. Geliş tarihi: 02.08.2013 / Kabul tarihi: 09.10.2013 Address for correspondance Latife ÇAKIR, DVM, PhD Associate Professor Department of Pathology Faculty of Veterinary Medicine University of Erciyes, 38039, Kayseri, Turkey e-mail: lcakir@erciyes.edu.tr
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
References
- Abbas B, Razig SA (1979): The etiology of keratoconjunctivitis occurring in goats in the Sudan. Vet Rec, 105, 348-350.
- Atkins KA, Powers CN ( 2002): The cytopathology of infectious diseases. Adv Anat Pathol, 9, 52-64.
- Bankemper KW, Lindley DM, Nusbaum KE, Mysinger RH (1990): Keratoconjunctivitis associated with Neisseria ovis infection in a herd of goats. J Vet Diagn Invest, 2, 76-78.
- Bauer AW, Kirby WMM, Sherris JC, Turck M (1966): Antibiotic susceptibility testing by a standardized single disk method. Am J Clin Pathol, 36, 493-496.
- Bauer GA, Spiess BM, Lutz H (1996): Exfoliative cytology of conjunctiva and cornea in domestic animals: a comparison of four collecting techniques. Vet Comp Ophthalmol, 6, 181-186.
- Belloy L, Janovsky M, Vilei EM, Pilo P, Giacomettim M, Frey J (2003): Molecular epidemiology of Mycoplasma conjunctivae in caprinae: Transmission across species in natural outbreaks. Appl Environ Microbiol, 69, 1913- 1919.
- Bolzan, A, Brunelli AJ, Castro M, Souza M, Souza J, Laus J (2005): Conjunctival impression cytology in dogs. Vet ophthalmol, 8, 401-405.
- Busch TJ, Belton DR (1988): Infectious keratoconjunctivitis in goats. N Z Vet J, 36, 153-155.
- Çakır L, Hızlısoy H, Abay S, Kutsal O (2011): Investigation of ocular surface cytology (brush-impression techniques) and Bacteriological Agents in dogs with Conjunctivitis and Healthy. Cytopathology, 22, 19.
- Dagnall GJ (1994): Use of exfoliative cytology in the diagnosis of ovine keratoconjunctivitis. Vet Rec, 135, 127- 130.
- Dubielzig RR, Ketring KL, McLellan GJ, Albert DM (2010): Veterinary Ocular Pathology A Comparative Review. 1st Edition. Saunders Elsevier, Oxford.
- Finger PT, Papp C, Latkany P, Kurli M, Iacob CE (2006):Anterior chamber paracentesis cytology (cytospin technique) for the diagnosis of intraocular lymphoma. Br J Ophthalmol, 6, 690-692.
- Fujihara T, Takeuchi T, Saito K, Kitajima Y, Kobayashi TK, Tsubato K (1997): Evaluation of Human Conjunctival Epithelium by a Combination of Brush Cytology and Flow Cytometry: An Approach to the Quantitative Technique. Diagn Cytopathol, 17, 456-460.
- Giacometti M, Janovsky M, Belloy L, Frey J (2002): Infectious keratoconjunctivitis of ibex, chamois and other Caprinae. Rev Sci Tech Off Int Épizoot, 21, 335–345.
- Gilger BC (2006): Ocular cytology-your key to immediate ocular diagnosis. 871-872. In: Proceedings of the North American Veterinary Conference, NAVG Gainesville, FL, USA.
- Hillström A, Tvedten H, Maria Källberg M, Hanås S, Lindhe A, Holst BS (2012): Evaluation of cytologic findings in feline conjunctivitis. Vet Clin Path, 41, 283–290.
- Jégou JP, Liotet S (1993): The benefit of conjunctival scraping cytology in the biological diagnosis of conjunctivitis in the dog and cat. EJCAP, 3, 83–97.
- Jones GE (1991): Infectious keratoconjunctivitis. 280– 283. In: Martin WB., Aitken ID., Editors: Diseases of sheep, Blackwell Scientific Publications.
- König CD (1983): Keratoconjunctivitis infectiosa ovis (KIO), 'pink eye' or 'zere oogjes' (a survey). Vet Q, 5, 127- 130.
- Lavach JD, Thrall MA, Benjamin MM, Severin GA (1977). Cytology of normal and inflamed conjunctivas in dogs and cats. J Am Vet Med Assoc, 170, 722–727.
- Levine TS, Rolfe KJ, Crow J, Styles S, Perrett CW, Maclean AB, Reid WMN (2001): The use of cytospin monolayer technique in the cytological diagnosis of vulval and anal disease. Cytopathology, 12, 297–305.
- Maggs DJ (2001): Laboratory investigation of ophthalmic disease. 24-29. In: Petersen-Jones SM, Crispin S (Eds), BSAVA Manual of Small Animal Ophthalmology. Second Edition. British Small Animal Veterinary Association, England.
- Marco I, Mentaberre G, Ballesteros C, Bischof DF, Lavín S, Vilei EM (2009): First Report of Mycoplasma conjunctivae from Wild Caprinae with Infectious Keratoconjunctivitis in the Pyrenees (NE Spain). J Wildlife Dis, 45, 238-241.
- Massa KL, Murphy CJ, Hartmann FA, Miller PE, Korsower CS, Young KM (1999): Usefulness of aerobic microbial culture and cytological evaluation of corneal specimens in the diagnosis of infectious ulcerative keratitis in animals. J Am Vet Med Assoc, 215,1671-1674.
- Mayer D, Nicolet J, Giacometti M, Schmitt M, Wahli T, Meier W (1996): Isolation of Mycoplasma conjunctivae from conjunctival swabs of Alpine ibex (Capra ibex ibex) affected with infectious keratoconjunctivitis. J Vet Med Ser B, 43, 155-161.
- Mushi EZ, Binta MG, Chabo RG, Dintwe K (2007): Conjunctival Flora of Fifty Healthy Goats in Sebele Farm, Gaborone, Botswana. J Anim Vet Adv, 6, 1388-1389.
- Prado MR, Rocha MFG, Brito ÉHS, Girão MD, Monterio AJ, Teixeira MFS, Sidrim JC (2005): Survey of bacterial microorganisms in the conjunctival sac of clinically normal dogs and dogs with ulcerative keratitis in Fortaleza, Ceará, Brazil. Vet Ophthalmol, 8, 33-37.
- Shanholtzer CJ, Schaper PJ, Peterson LR (1982): Concentrated gram stain smears prepared with a cytospin centrifuge. J Clin Microbiol, 16, 1052–1056.
- Sharma S (2012): Diagnosis of infectious diseases of the eye. Eye, 26,177-84.
- Takano Y, Fukagawa K, Dogru M, Asano-Kato N, Tsubota K, and Fujishima H (2004): Inflammatory cells in brush cytology samples correlate with the severity of corneal lesions in atopic keratoconjunctivitis. Br J Ophthalmol, 88, 1504–1505.
- Ter Laak EA, Schreuder BE, Smith-Buys CM (1998): The occurance of Mycoplasma conjunctivae in the Netherlands and its association with infectious kerato- conjunctivitis in sheep and goats. Vet Quart, 10, 73-83.
- Tschopp R, Frey J, Zimmermann L, Giacometti M (2005): Outbreaks of infectious keratoconjunctivitis in alpine chamois and ibex in Switzerland between 2001 and 2003. Vet Rec, 157,13-18.
- Uchida Y (1990): Viral diseases of the outer eye–rapid diagnosis by immunohistochemistry. Nippon Ganka Gakkai Zasshi, 94, 889–902.
- Wakamatsu TH, Okada N, Kojima T, Matsumoto Y, Ibrahim, OMA, Adan ES, Fukagawa K, Katakami C, Tsubota K, Shimazaki J, Fujishima H (2009): Evaluation of conjunctival inflammatory status by confocal scanning laser microscopy and conjunctival brush cytology in patients with atopic keratoconjunctivitis (AKC). Mol Vis, 15, 1611–1619.
- Willis M, Bounous DI, Hirsh S, Kaswan R, Stiles J, Martin C, Rakich P, Roberts W (1997): Conjunctival brush cytology: evaluation of a new cytological collection technique in dogs and cats with a comparison to conjunctival scraping. Vet Comp Ophthalmol, 7, 74-81.
- Yağmur M, Ersöz C, Ersöz TR, Varinli S (1997): Brush technique in ocular surface cytology. Diagn Cytopathol, 17, 88-91. Geliş tarihi: 02.08.2013 / Kabul tarihi: 09.10.2013 Address for correspondance Latife ÇAKIR, DVM, PhD Associate Professor Department of Pathology Faculty of Veterinary Medicine University of Erciyes, 38039, Kayseri, Turkey e-mail: lcakir@erciyes.edu.tr
Details
| Primary Language | English |
|---|---|
| Subjects | Veterinary Surgery |
| Other ID | JA58TM98BV |
| Journal Section | Research Article |
| Authors | |
| Publication Date | March 1, 2014 |
| Published in Issue | Year 2014Volume: 61 Issue: 1 |