Abstract
The aim of this study is to determine C. parvum in small intestinal samples of lambs (up to 30 days old) with enteritis. For this aim, fresh intestinal smears prepared from 60 lambs (euthanized due to diarrhea) and stained with Modified Ziehl-Neelsen (MZN). In addition to these 60 fresh samples, 85 different small intestinal samples obtained from dead lambs (total 145 small intestinal samples) were embedded in paraffin for histopathology, immunofluorescence (IF) and immunohistochemistry (IHC). Slides were stained according to protocols (MZN, hematoxylin-eosin, IHC and IF). In MZN staining, 13.3% of smear samples were found positive for Cryptosporidium spp. Besides, C. parvum positivity was detected in the rate of 13.8% in IF and IHC stains. In the histopathologic examination, desquamation and necrosis of intestinal epithelium, villus atrophy, Cryptosporidium spp. along the surface of villus epithelium, hyperemia, mononuclear cellular infiltration, mitosis in crypt epithelium, intraepithelial lymphocytes, dilatation in the lymphatic canal and Eimeria spp. in some cases were observed in varying degree. In conclusion, IF, IHC and MZN stains were found effective in revealing cryptosporidiosis in lambs.
Keywords
enteritis histopathology immunofluorescence C. parvum Cryptosporidiosis immunohistochemistry
References
- 1. Ahourai P, Ezzi A, Gholami MR, et al. (1985): Cryptosporidium spp. in new born lambs in Iran. Trop Anim Health Prod, 17, 6-8.
- 2. Angus KW, Tzipori S, Gray EW (1982): Intestinal lesions in specific-pathogen-free lambs associated with a Cryptosporidium from calves with diarrhea. Vet Pathol, 19,67-68.
- 3. Arrowood MJ, Sterling CR (1989): Comparison of conventional staining methods and monoclonal antibody-based methods for Cryptosporidium oocyst detection. J Clinical Microbiol, 27, 1490-5.
- 4. Aytuğ N, Alaçam E, Özkoç Ü (1990) Koyun-Keçi Hastalıkları ve Yetiştiriciliği, TÜM VET Hayvancılık Hizmetleri, İstanbul, 212-213.
- 5. Bejan A, Bolfa P, Magdas C (2011): Morphological, histopathological and immunofluorescence characterization of Cryptosporidium parvum natural infection in goat kids from Romania. Acta Vet, 61, 609-619.
- 6. Bilal T, Bilal T (2005): Koyun-Keçilerin İç Hastalıkları ve Beslenmesi. İstanbul Üniversitesi Basım ve Yayınevi Müdürlüğü, İstanbul, 17-21.
- 7. Brown CC, Baker DC, Barker IK (2007): Alimentary System. 274-277. In: MG Maxie (ed) Jubb Kennedy and Palmer’s Pathology of Domestic Animals. Saunders/Elsevier, Philadelphia.
- 8. Casuape AC, Quilez J, Acedo CS (2002): Prevalence and analysis of potential risk factors for Cryptosporidium parvum infection in lambs in Zaragoza (northeastern Spain). Vet Parasitol, 104, 287-98.
- 9. Coop RL,Wright SE (2003): Cryptosporidiosis and Coccidiosis. 153-155 In: WB Martin, ID Aitken (ed) Diseases of Sheep. 3th ed, Blackwell Science, Philadelphia, USA, 2003.
- 10. Erman N, Beyazıt A, Oz I (2000): Prevalence of cryptosporidiosis in lambs and goat kids in İzmir province. Bornova Vet Kont Araş Enst Derg, 25, 33-38.
- 11. Fayer L, Xiao L (2008): Cryptosporidium and Cryptosporidiosis, Taylor and Francis Group, Florida.
- 12. Gökçe E, Erdoğan HM (2009): An epidemiological study on neonatal lamb health. Kafkas Üniv Vet Fak Derg, 15, 225-236.
- 13. Gökçe E, Ünver A, Erdoğan HM (2010) İshalli neonatal kuzularda enterik patojenlerin belirlenmesi. Kafkas Üniv Vet Fak Derg, 16, 717-722.
- 14. İnci A (2013): Sığırlarda Cryptosporidiosis. 135-142 In: MA Özcel (ed). Veteriner Hekimliğinde Parazit Hastalıkları, Meta Basım Matbaacılık Hizmetleri, İzmir.
- 15. Khurana S, Sharma P, Sharma A et al. (2012): Evaluation of Ziehl-Neelsen staining, auramine phenol staining, antigen detection enzyme linked immunosorbent assay and polymerase chain reaction, for the diagnosis of intestinal cryptosporidiosis. Trop Parasitol, 2, 20-23.
- 16. Loose JH, Sedergran DJ, Cooper HS (1989): Identification of cryptosporidium in paraffin-embedded tissue sections with the use of a monoclonal antibody. American J Clinical Pathology, 91, 206-209.
- 17. Milli ÜH (1997): Sindirim Sistemi. 132-134, In: ÜH Milli, R Hazıroğlu (ed) Veteriner Patoloji, Tamer Matbaacılık, Ankara.
- 18. Munoz M, Alvarez M, Lanza I et al. (1996): Role of enteric pathogens in the aetiology of neonatal diarrhoea in lambs and goat kids in Spain. Epidemiology & Infection, 117, 203-211.
- 19. Naciri M, Paul Lefay MP, Mancassola R, et al. (1999): Role of Cryptosporidium parvum as a pathogen in neonatal diarrhoea complex in suckling and dairy calves in France. Vet Parasitol, 85, 245-247.
- 20. OIE Terrestrial Manual (2015). Cryptosporidiosis. http://www.oie.int/fileadmin/Home/eng/Health_standards/tahm/2.09.04_CRYPTO.pdf (02 Mayıs 2015).
- 21. Ozdal N, Tanrıtanır P, Göz Y et al. (2009): Parasitic protozoans (Eimeria, Giardia, and Cryptosporidium) in lambs with diarhoea in the Van province. Bull Vet Inst Pulawy, 53, 47-51.
- 22. Ozer E, Erdogmus SZ, Koroglu E (1990). Investigation on the incidence of Cryptosporidia of calves and lambs in Elazig vicinity. Turk J Vet Anim Sci, 14, 439–445.
- 23. Özmen Ö, Serpin N, Jamshidi K (2016): Kuzu ve oğlaklarda kriptosporidioziste patolojik ve immunohistokimyasal bulgular. MAE Vet Fak Derg, 1, 7-13.
- 24. Perkins ME, Riojas YA, Wu TW et al. (1999): CpABC, a Cryptosporidium parvum ATP-binding cassette protein at the host-parasite boundary in intracellular stages. Proc Natl Acad Sci USA, 96, 5734–5739.
- 25. Quílez J, Sánchez-Acedo C, Clavel A et al. (1996): Comparison of an acid-fast stain and a monoclonal antibody-based immunofluorescence reagent for the detection of Cryptosporidium oocysts in faecal specimens from cattle and pigs. Vet Parasitol, 67, 75-81.
- 26. Rusnak J, Hadfield TL, Rhodes MM et al. (1989): Detection of Cryptosporidium oocysts in human fecal specimens by an indirect immunofluorescence assay with monoclonal antibodies. J Clinical Microbiol, 27,1135-6.
- 27. Sari B, Arslan MO, Gicik Y et al. (2009): The prevalence of Cryptosporidium species in diarrhoeic lambs in Kars province and potential risk factors. Trop Anim Health Prod, 41, 819-826.
- 28. Sevinc F, Irmak K, Sevinç M (2003): The prevalence of Cryptosporidium parvum infection in the diarrhoiec and non- diarrhoeic calve. Rev Med Vet, 5,357-361.
- 29. Sevinç F, Dik B (2010): Cryptosporidiae. 155-161, In: N Dumanlı, Z Karaer (ed) Veteriner Protozooloji, 1. Baskı, Medisan, Ankara.
- 30. Sevinç F, Uslu U, Derinbay Ö (2005): The prevalance of Cryptosporidium parvum in lambs around Konya. Turk J Vet Animal Sci, 29, 1191-1194.
- 31. Stibbs HH, Ongerth JE (1986): Immunofluorescence detection of Cryptosporidium oocysts in fecal smears. J Clinical Microbiol, 24, 517-21.
- 32. Ulutaş B, Voyvoda H (2004): Cryptosporidiosis in diarrhoeic lambs on a sheep farm. Turkiye Parazitol Derg, 28,15-17.
Abstract
Çalışmanın amacı kuzu ishallerinde önemli bir patojen olan Cryptosporidium parvum (C. parvum) etkenlerinin bağırsak smearı ve parafin kesitlerden, Modifiye Ziehl-Neelsen (MZN), histopatoloji, immunohistokimya (İHK) ve immunfloresan (İF) boyama yöntemleri ile belirlemek ve Cryptosporidiosis hastalığının 30 günlüğe kadar olan kuzulardaki yaygınlığını ortaya koymaktır. Bu amaçla, ishal belirtisi gösteren ve agoni halde iken nekropsi yapılmış 60 adet ve yine ishal belirtisiyle ölmüş 85 adet (toplam 145 adet) kuzudan alınan bağırsak örnekleri kullanılmıştır. MZN boyama sadece, agoni halde iken nekropsileri yapılan 60 kuzudan alınan taze incebağırsak örneklerinden hazırlanan smear örneklerine yapılmıştır. Bu 60 örnekle birlikte ölü olarak getirilen 85 kuzudan (toplam 145) elde edilen incebağırsak örneklerinden hazırlanan parafin kesitlerden ise histopatoloji, İF ve İHC boyamaları prosedürüne uygun olarak yapıldı. MZN boyamalarda (60 örnek) %13.3 oranında Cryptosporidium spp., parafin blokları hazırlanan toplam 145 örnekten alınan kesitlerin IF ve IHC boyamalarda ise %13.8 oranında C. parvum pozitif reaksiyon gözlenmiştir. Etken tespit edilen bağırsak kesitlerinin histopatolojik incelemelerinde; değişen derecelerde olmak üzere; epitel hücrelerinde nekroz, deskuamasyon ve villuslarda kütleşme, epitel yüzeyi boyunca Cryptosporidium spp. etkenleri, propria mukozada hiperemi ve mononükleer hücre infiltrasyonu, kript epitelinde mitoz artışı, lenfatik kanalda genişleme, intraepitelyal lenfositler ve bazı örneklerde Eimeria spp gözlendi. Sonuç olarak çalışmada İF, İHC ve MZN boyamalarının kuzularda Cryptosporidiosis hastalığını ortaya koymada etkili olduğu görülmüştür.
References
- 1. Ahourai P, Ezzi A, Gholami MR, et al. (1985): Cryptosporidium spp. in new born lambs in Iran. Trop Anim Health Prod, 17, 6-8.
- 2. Angus KW, Tzipori S, Gray EW (1982): Intestinal lesions in specific-pathogen-free lambs associated with a Cryptosporidium from calves with diarrhea. Vet Pathol, 19,67-68.
- 3. Arrowood MJ, Sterling CR (1989): Comparison of conventional staining methods and monoclonal antibody-based methods for Cryptosporidium oocyst detection. J Clinical Microbiol, 27, 1490-5.
- 4. Aytuğ N, Alaçam E, Özkoç Ü (1990) Koyun-Keçi Hastalıkları ve Yetiştiriciliği, TÜM VET Hayvancılık Hizmetleri, İstanbul, 212-213.
- 5. Bejan A, Bolfa P, Magdas C (2011): Morphological, histopathological and immunofluorescence characterization of Cryptosporidium parvum natural infection in goat kids from Romania. Acta Vet, 61, 609-619.
- 6. Bilal T, Bilal T (2005): Koyun-Keçilerin İç Hastalıkları ve Beslenmesi. İstanbul Üniversitesi Basım ve Yayınevi Müdürlüğü, İstanbul, 17-21.
- 7. Brown CC, Baker DC, Barker IK (2007): Alimentary System. 274-277. In: MG Maxie (ed) Jubb Kennedy and Palmer’s Pathology of Domestic Animals. Saunders/Elsevier, Philadelphia.
- 8. Casuape AC, Quilez J, Acedo CS (2002): Prevalence and analysis of potential risk factors for Cryptosporidium parvum infection in lambs in Zaragoza (northeastern Spain). Vet Parasitol, 104, 287-98.
- 9. Coop RL,Wright SE (2003): Cryptosporidiosis and Coccidiosis. 153-155 In: WB Martin, ID Aitken (ed) Diseases of Sheep. 3th ed, Blackwell Science, Philadelphia, USA, 2003.
- 10. Erman N, Beyazıt A, Oz I (2000): Prevalence of cryptosporidiosis in lambs and goat kids in İzmir province. Bornova Vet Kont Araş Enst Derg, 25, 33-38.
- 11. Fayer L, Xiao L (2008): Cryptosporidium and Cryptosporidiosis, Taylor and Francis Group, Florida.
- 12. Gökçe E, Erdoğan HM (2009): An epidemiological study on neonatal lamb health. Kafkas Üniv Vet Fak Derg, 15, 225-236.
- 13. Gökçe E, Ünver A, Erdoğan HM (2010) İshalli neonatal kuzularda enterik patojenlerin belirlenmesi. Kafkas Üniv Vet Fak Derg, 16, 717-722.
- 14. İnci A (2013): Sığırlarda Cryptosporidiosis. 135-142 In: MA Özcel (ed). Veteriner Hekimliğinde Parazit Hastalıkları, Meta Basım Matbaacılık Hizmetleri, İzmir.
- 15. Khurana S, Sharma P, Sharma A et al. (2012): Evaluation of Ziehl-Neelsen staining, auramine phenol staining, antigen detection enzyme linked immunosorbent assay and polymerase chain reaction, for the diagnosis of intestinal cryptosporidiosis. Trop Parasitol, 2, 20-23.
- 16. Loose JH, Sedergran DJ, Cooper HS (1989): Identification of cryptosporidium in paraffin-embedded tissue sections with the use of a monoclonal antibody. American J Clinical Pathology, 91, 206-209.
- 17. Milli ÜH (1997): Sindirim Sistemi. 132-134, In: ÜH Milli, R Hazıroğlu (ed) Veteriner Patoloji, Tamer Matbaacılık, Ankara.
- 18. Munoz M, Alvarez M, Lanza I et al. (1996): Role of enteric pathogens in the aetiology of neonatal diarrhoea in lambs and goat kids in Spain. Epidemiology & Infection, 117, 203-211.
- 19. Naciri M, Paul Lefay MP, Mancassola R, et al. (1999): Role of Cryptosporidium parvum as a pathogen in neonatal diarrhoea complex in suckling and dairy calves in France. Vet Parasitol, 85, 245-247.
- 20. OIE Terrestrial Manual (2015). Cryptosporidiosis. http://www.oie.int/fileadmin/Home/eng/Health_standards/tahm/2.09.04_CRYPTO.pdf (02 Mayıs 2015).
- 21. Ozdal N, Tanrıtanır P, Göz Y et al. (2009): Parasitic protozoans (Eimeria, Giardia, and Cryptosporidium) in lambs with diarhoea in the Van province. Bull Vet Inst Pulawy, 53, 47-51.
- 22. Ozer E, Erdogmus SZ, Koroglu E (1990). Investigation on the incidence of Cryptosporidia of calves and lambs in Elazig vicinity. Turk J Vet Anim Sci, 14, 439–445.
- 23. Özmen Ö, Serpin N, Jamshidi K (2016): Kuzu ve oğlaklarda kriptosporidioziste patolojik ve immunohistokimyasal bulgular. MAE Vet Fak Derg, 1, 7-13.
- 24. Perkins ME, Riojas YA, Wu TW et al. (1999): CpABC, a Cryptosporidium parvum ATP-binding cassette protein at the host-parasite boundary in intracellular stages. Proc Natl Acad Sci USA, 96, 5734–5739.
- 25. Quílez J, Sánchez-Acedo C, Clavel A et al. (1996): Comparison of an acid-fast stain and a monoclonal antibody-based immunofluorescence reagent for the detection of Cryptosporidium oocysts in faecal specimens from cattle and pigs. Vet Parasitol, 67, 75-81.
- 26. Rusnak J, Hadfield TL, Rhodes MM et al. (1989): Detection of Cryptosporidium oocysts in human fecal specimens by an indirect immunofluorescence assay with monoclonal antibodies. J Clinical Microbiol, 27,1135-6.
- 27. Sari B, Arslan MO, Gicik Y et al. (2009): The prevalence of Cryptosporidium species in diarrhoeic lambs in Kars province and potential risk factors. Trop Anim Health Prod, 41, 819-826.
- 28. Sevinc F, Irmak K, Sevinç M (2003): The prevalence of Cryptosporidium parvum infection in the diarrhoiec and non- diarrhoeic calve. Rev Med Vet, 5,357-361.
- 29. Sevinç F, Dik B (2010): Cryptosporidiae. 155-161, In: N Dumanlı, Z Karaer (ed) Veteriner Protozooloji, 1. Baskı, Medisan, Ankara.
- 30. Sevinç F, Uslu U, Derinbay Ö (2005): The prevalance of Cryptosporidium parvum in lambs around Konya. Turk J Vet Animal Sci, 29, 1191-1194.
- 31. Stibbs HH, Ongerth JE (1986): Immunofluorescence detection of Cryptosporidium oocysts in fecal smears. J Clinical Microbiol, 24, 517-21.
- 32. Ulutaş B, Voyvoda H (2004): Cryptosporidiosis in diarrhoeic lambs on a sheep farm. Turkiye Parazitol Derg, 28,15-17.
Details
| Primary Language | Turkish |
|---|---|
| Subjects | Veterinary Surgery |
| Journal Section | Research Article |
| Authors | |
| Publication Date | April 5, 2019 |
| Published in Issue | Year 2019Volume: 66 Issue: 2 |