Comparison between diagnostic methods of Cryptosporidium spp. in fecal samples from cattle

Authors

  • Bruna Baccega Dpto de Microbiologia e Parasitologia da Universidade Federal de Pelotas
  • Pedro de Souza Quevedo Instituto de Estudos do Trópico Úmido da Universidade Federal do Sul e Sudeste do Pará
  • Juliana Montelli Fenalti Instituto de Estudos do Trópico Úmido da Universidade Federal do Sul e Sudeste do Pará
  • Cibele Veleda dos Santos Dpto de Microbiologia e Parasitologia da Universidade Federal de Pelotas
  • Nara Amélia da Rosa Farias Dpto de Microbiologia e Parasitologia da Universidade Federal de Pelotas
  • Elisa Simone Viegas Sallis Dpto de Microbiologia e Parasitologia da Universidade Federal de Pelotas

Keywords:

Cryptosporidiosis, parasitological diagnosis, protozoa, auramina coloration

Abstract

Cryptosporidiosis is an enteric disease with varied clinical manifestations and eventual mortality, mainly in young animals, causing impairment in development. This study aimed to compare staining techniques for the detection of Cryptosporidium spp. oocysts in dairy calf feces from 22 municipalities of the southern region of Rio Grande do Sul, Brazil. Fecal samples were collected directly from the rectum of 359 calves of different breeds, male and female, up to twelve months old. Cryptosporidium spp. oocysts were observed using the colorimetric methods of Kinyoun, Safranina and Ziehl-Neelsen and were visualized under optical microscopy and by fluorescence with auramine stain. Cryptosporidium spp. oocysts were observed in 6.69% (24/359) of the analyzed samples. From these results it can be inferred that the four colorimetric methods were effective in the detection of Cryptosporidium spp., being able to reveal this parasite even in samples

 

with a reduced number of oocysts. There was no statistically significant difference between the methods for diagnosis, even though, the Auramine staining fluorescence method presented the best result in comparison with the colorimetric techniques used in this study.

References

ARAÚJO, F.A.P.; WILSMANN, C.G.F.; TEIXEIRA, M.C. In: Enfermidades parasitárias por protozoários em pequenos animais/organização, Cláudia de Mello Ribeiro. 1ª ed., Rio de Janeiro: Rubio, 2015. 168p.

ARROWOOD, M.J. Diagnosis. In: Cryptosporidium and Cryptosporidiosis, Fayer R. 1a ed., CRC Press, New York, 1997. 576p.

BAXBY, D.; BLUNDELL, N.; HART, C.A. The development and performance of a simple sensitive method for the detection of Cryptosporidium oocysts in faeces. The Journal of Hygiene, v.93, p.317-323, 1983.

BAXBY, D.; HART, C.A.; BLUNDELL, N. Shedding of oocysts by immunocompetent individuals with cryptosporidiosis. The Journal of Hygiene, v.95, p.703-709, 1985.

BOWMAN, D.D.; LUCIO-FORSTER, A. Cryptosporidiosis and giardiasis in dogs and cats: veterinary and public health importance. Experimental Parasitology, v.124, p.121- 127, 2010.

BROGLIA, A.; RECKINGER, S.; CACCIÓ, S.M.; NÖCKLER, K. Distribution of Cryptosporidium parvum subtypes in calves in Germany. Veterinary Parasitology, v.154, p.8-13, 2008.

BROOK, E.; HART, C.A.; FRENCH, N.; CHRISTLEY, R. Prevalence and risk factors for Cryptosporidium spp. infection in young calves. Veterinary Parasitology, v.152, p.46-52, 2008.

CHAKO, C.Z.; TYLER, J.W.; SCHULTZ, L.G.; CHIGUMA, L.; BEERNTSEN, B.T. Cryptosporidiosis in people: It’s not just about the cows. Journal of Veterinary Internal Medicine, v.24, p.37-43, 2010.

CHALMERS, R.M.; SMITH, R.; ELWIN, K.; CLIFTON-HADLEY, F.A.; GILES, M. Epidemiology of anthroponotic and zoonotic human cryptosporidiosis in England and Wales, 2004–2006. Epidemiology and Infection, v.139, p.700-712, 2001.

DA CUNHA, M.J.R.; SILVA, M.B.O.; ARANTES, V.M.; CURY, M.C. Comparação da eficácia do Ziehl-Neelsen e da Reação em Cadeia de Polimerase (PCR) para o diagnóstico do Cryptosporidium spp. em bovinos e suínos provenientes da região de Uberlândia. In: Anais do XXI Congresso Brasileiro de Parasitologia, 2009, Foz do Iguaçu. 2009.

DAS, G.; CHANGKIJA, B.; SARKAR, S.; DAS, P. Genotyping of Cryptosporidium parvum isolates in bovine population in Kolkata and characterization of new bovine genotypes. Research in Veterinary Science, v.91, p.246-250, 2011.

DE CARLI, G.A. Diagnóstico laboratorial das parasitoses humanas –Métodos e Técnicas. 1ª ed., Rio de Janeiro: Editora Médica e Científica Ltda, 1994. 313p.

DIAZ-LEE, A.; MOLINA, R.; DOUGNAC, C.; MERCADOB, R.; RETAMALA, P.; FREDESA, P. Sensibilidad analítica de técnicas de tinción tradicionales y una técnica molecular para la detección de ooquistes de Cryptosporidium spp. aislados de bovinos en muestras de agua: estudio preliminar. Archive Medicine Veterinary, v.47, p.91-96, 2015.

FALL, A.; THOMPSON, R.C.A.; HOBBS, R.P.; MORGAN-RYAN, U. Morphology is not a reliable tool for delineating species within Cryptosporidium. Journal of Parasitology, v.89, p.399-402, 2003.

FAYER, R.; MORGAN, U.; UPTON, S.J. Epidemiology of Cryptosporidium: transmission, detection and identification. International Journal of Parasitology, v.30, p.1305-1322, 2000.

FAYER, R.; SANTIN, M.; TROUT, J.M. Cryptosporidium ryanae n. sp (Apicomplex: Cryptosporidiidae) in cattle (Bos taurus). Veterinary Parasitology, v.156, p.191–190, 2008.

FEITOSA, F.L.; SHIMAMURA, G.M.; ROBERTO, T.; MEIRELES, M.V.; NUNES, C.M.; CIARLINI, P.C.; BORGES, A.S. Prevalência de criptosporidiose em bezerros na região de Araçatuba, estado de São Paulo, Brasil. Ciência Rural, v.34, p.189-193, 2004.

GARCIA, L.S.; BRUCKNER, D.A.; BREWERT, T.C.; SMINITZU, R.Y. Techniques for the recovery and identification of Cryptosporidium oocysts from stoll specimens. Journal Clinical of Microbiology, v.18, p.185-190, 1983.

GRACZYK, T.K.; MAJEWSKA, A.C.;

SSCHWAB, K.J. The role of birds in

dissemination of human waterborne enteropathogens. Trends in parasitology, v.24, p.55- 59, 2008.

HENDERSON, H.J.; SPAULDING, E.H.; GAULT, E.S. Demonstration of globi and leprosy bacilli by fluorescence microscopy. Proceedings of the Society for Experimental Biology and Medicine, v.50, p.91-2, 1942.

IGNATIUS, R.; EISENBLATTER, M.; REGNATH, T.; MANSMANN, U.; FUTH, U.;

HAHN, H.; WAGNER, J. Efficacy of different methods for detection of low Cryptosporidium parvum oocyst numbers or antigen concentrations in stool specimens- Eur. Journal Clinical Microbiology Infections Diseases, v.16, p.732-6, 1997.

KHAN, S.M.; DEBNATH, C.; PRAMANIK, A.K.; XIAO, L.; NOZAKI, T.; GANGULY,

S. Molecular characterization and assessment of zoonotic transmission of Cryptosporidium from dairy cattle in West Bengal, India. Veterinary Parasitology, Amsterdam, v.171, p.41- 47, 2010.

KHEL, K.S.C.; CICIRELLO, H.; HAVENS, P.L. Comparison of four different methods for detection of Cryptosporidium species. Journal of Clinical Microbiology, v.33, p.416- 418, 1995.

LANGONI, H.; LINHARES, A.C.; AVILA, F.A.; SILVA, A.V. DA; ELIAS, A.O.

Contribution to the study of diarrhea etiology in neonate dairy calves in São Paulo state, Brazil. Brazilian Journal of Veterinary Research and Animal Science, v.41, p.313-319, 2004.

MACEDO, R. Detecção e caracterização molecular de picobirnavirus em fezes de bezerros naturalmente infectados. 65p. (Dissertação) Paraná, Universidade Federal do Paraná, 2014.

MEIRELES, M.V.; OLIVEIRA, F.P.; TEIXEIRA, W.F.P.; COELHO, W.M.D.; MENDES,

L.C.N. Molecular characterization of Cryptosporidium spp. in dairy calves from the state of São Paulo, Brazil. Parasitology Research, v.109, p.949-951, 2011.

MIRHASHEMI, M.E.; ZINTL, A.; GRANT, T.; LUCY, F. E.; MULCAHY, G.; DE

WAAL, T. Comparison of diagnostic techniques for the detection of Cryptosporidium oocysts in animal samples. Experimental Parasitology, v.151, p.14-20, 2015.

MOURA, H.; OLIVEIRA, L.M. Cryptosporidium: parasito de imunocomprometidos. Revista Brasileira Patologia Clínica, v.21, p.198-201, 1985.

MUNDIN, M.J.S.; SOUZA, L.M. DE; MUNDIM, A.V.; MORAIS, R.N. Frequência de

oocistos de Cryptosporidium sp em fezes de bezerros criados sob condições naturais no município de Uberlândia, analisadas por quatro métodos laboratoriais. Veterinária Notícias, v.1, p.33-36, 1995.

MUNÕZ, P.; FREDES, F.; DIAZ-LEE, A.; MERCADO, R.; OZAKI, L. Deteccion de

Cryptosporidium spp. em terneras de lecherias de la Region Metropolitana mediante Ziehl Neelsen y confirmada por inmunocromatografia y ensayo molecular. Archive Medicine Veterinary, v.43, p.111-116, 2011.

NAZEMALHOSSEINI-MOJARAD, E.; HAGHIGHI, A.; TAGHIPOUR, N.; KESHAVARZ, A.; MOHEBI, S.R.; ZALI, M.R.; XIAO, L. Subtype analysis of

Cryptosporidium parvum and Cryptosporidium hominis isolates from humans and cattle in Iran. Veterinary Parasitology, v.179, p.250-252, 2011.

NETA, E.S.M.; SAMPAIO, D.C.; GALVÃO, G.S.; MUNHOZ, A.D. Oocistos de

Cryptosporidium (Apicomplexa: cryptosporidiidae) em bovinos leiteiros de uma área endêmica na Microrregião de Ilhéus-Itabuna, Estado da Bahia, Brasil. Revista Brasileira de Medicina Veterinária, v.32, p.75-78, 2010.

PANCIERA, R.J.; THOMASSEN, R.W.; GARNER, F.M. Cryptosporidial infection in a calf. Veterinary Pathology, v.8, p.479-484, 1971.

QUADROS, R.M.D.; ARAÚJO, F.A.P. Ocorrência de Cryptosporidium sp. Tyzzer, 1907 detectada pelo método de imunofluorescência através da técnica de coloração da auramina em bovinos em propriedades rurais do município de Lages (SC), Brasil. Revista de Ciência Agro veterinária, v.2, p.68-73, 2003.

QUADROS, R.M.D.; MARQUES, S.M.T.; AMENDOEIRA, C.R.; SOUZA, L.A.;

AMENDOEIRA, P.R.; COMPARIN, C.C. Detection of Cryptosporidium oocysts by auramine and Ziehl-Neelsen staining methods. Parasitologia Latinoamericana, v.61, p.117- 120, 2006.

REY, L. Parasitologia. 2ªed., Guanabara Koogan, Rio de Janeiro.1991. 731p.

RIBEIRO, M.G.; LANGONI, H.; JEREZ, J.A.; LEITE, D.S.; FERREIRA, F.; GENNARI,

S.M. Identification of enteropathogens from buffalo calves with and without diarrhoea in the Ribeira Valley, State of São Paulo, Brazil. Brazilian Journal of Veterinary Research and Animal Science, v.37, p.34-39, 2000.

RIBEIRO, C.M.; KATAGIRI, S. Criptosporidiose. In: Enfermidades parasitárias por protozoários em pequenos animais/organização, Cláudia de Mello Ribeiro, 1ª ed., Rio de Janeiro: Rubio, 2015. 168p.

RITCHIE, L.S. An ether sedimentation technique for routine stool examination. Bulletin of the United States Army Medical Department, v.8, p.326, 1948.

ROBERTSON, L.J.; CHALMERS, R.M. Foodborne cryptosporidiosis: is there really more in Nordic countries? Trends in parasitology, v.29, p.3-9, 2013.

RODRIGUES, R.D.; GOMES, L.R.; SOUZA, R.R.; BARBOSA, F.C. Comparação da

eficiência das colorações de Ziehl-Neelsen modificado e Safranina modificada na detecção de oocistos de Cryptosporidium spp. (Eucoccidiorida, Cryptosporidiidae) a partir de amostras fecais de bezerros de 0 a 3 meses. Ciência Animal Brasileira, v.17, n.1, p.119- 125, 2016.

RYAN, U.; FAYER, R.; XIAO, L. Cryptosporidium species in humans and animals: current understanding and research needs. Parasitology, v.141, p.1667-1685, 2014.

SILVERLAS, C.; EMANUELSON, U.; VERDIER, K.; BJORKMAN, C. Prevalence and

associated management factors of Cryptosporidium shedding in 50 Swedish dairy herds. Preventive Veterinary Medicine, v.90, p.242-253, 2009.

TAYLOR, M.A.; WEBSTER, K.A. Recent advances in the diagnosis in livestock of Cryptosporidium, Toxoplasma, Giardia and other protozoa of veterinary importance. Research and Veterinary Science, v.65, p.83-193, 1998.

TEIXEIRA, W.F.P.; COELHO, W.M.D.; NUNES, C.M.; MEIRELES, M.V. Detection of

Cryptosporidium parvum oocysts in calf fecal samples by direct immunofluorescence assay. Revista Brasileira de Parasitologia Veterinária, v.20, n.4, p.269-273, 2011.

THOMPSON, R.C.; PALMER, C.S.; O'HANDLEY, R. The public health and clinical significance of Giardia and Cryptosporidium in domestic animals. Veterinary Journal, v.177, p.18-25, 2008.

XIAO, L.; HERD, R.P. Infection pattern of Cryptosporidium and Giardia in calves. Veterinary Parasitology, v.55, p.257-62, 1993.

XIAO, L. Molecular epidemiology of cryptosporidiosis: An update. Experimental Parasitology, v.124, p.80-89, 2010.

YODER, J.S.; WALLACE, R.M.; COLLIER, S.A.; BEACH, M.J.; HLAVSA, M.C.

Cryptosporidiosis surveillance United States, 2009–2010. CENTERS FOR DISEASE, C.; PREVENTION. The Morbidity and Mortality Weekly Report Surveillance Summaries, v.61, p.1-12, 2012.

YOUNG, K.H.; BULLOCK, S.L.; MELVIN, D.M.; SPRUILL, C. L. Ethyl Acetat as a

substitute for Diethyl e Ether in the Formalin-Ether Sedimentation Tecnique. Journal Clinical Microbiology, v.10, p.852-853, 1979.

Published

2020-12-31

How to Cite

BACCEGA, B.; QUEVEDO, P. de S. .; FENALTI, J. M. .; SANTOS, C. V. dos .; FARIAS, N. A. da R.; SALLIS, E. S. V. . Comparison between diagnostic methods of Cryptosporidium spp. in fecal samples from cattle. Ciência Animal, [S. l.], v. 30, n. 1, p. 36–46, 2020. Disponível em: https://revistas.uece.br/index.php/cienciaanimal/article/view/9650. Acesso em: 21 nov. 2024.

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