STANDARDIZATION OF EGG ECLOSION TEST: IN VITRO DIAGNOSIS OF RESISTANCE TO ALBENDAZOLE IN GASTROINTESTINAL NEMATODES
Abstract
Current analysis standardized, adapted and detailed egg eclosion test (EET) to diagnose the resistance of gastrointestinal nematodes to albendazole in sheep. Three in vitro assays were undertaken at 24 and 48 h, by methodology prescribed by the EMBRAPA Practical Handbook, with negative control distilled water and DMSO at 0.75% and concentration at 600, 300, 150, 112.5, 75, 37.5, 18.75, 9.37 and 4.68 μg/mL and positive control albendazole 25.000 μg/mL. Plates were incubated in buffer with BOD and maintained at 28ºC. After 24h and 48h of incubation, larvae had eclosion in negative control groups and also in groups treated with 112.5 μg/mL and 75 μg/mL, respectively. It was not possible to diagnose anti-helminth resistance in EET due to limiting variables, such as the lack of discriminating dose for albendazole. The standardization, adaptation and detailing were possible to diagnose anti-helminth resistance of nematodes to albendazole.References
ALBONICO, M. et al. Development of the egg hatch assay for detection of anthelminthic resistance in human hookworms. International Journal for Parasitology, v.35, n.7, p. 803-811, 2005.
AMARANTE, A. F. T. et al. Resistance of Santa Inês, Suffolk and Ile de France sheep to naturally acquired gastrointestinal nematode infections. Veterinary Parasitololy, v.120, n.2, p.91-106, 2004.
BIZIMENYERA, E. S. et al. E. In vitro activity of Peltophorum africanum Sond. (Fabaceae) extracts on the egg hatching and larval development of the parasitic nematode Trichostrongylus colubriformis. Veterinary Parasitology, v.142, n.3, p.336-343, 2006.
BRASIL, Portaria SDA nº 48, de 12 de maio de 1997. Aprova como anexo o Regulamento Técnico a ser observado na produção, no controle e no emprego de antiparasitários de uso veterinário. In: Legislação relacionada aos produtos de uso veterinário. Brasília, 2012.
CHAGAS, A.C.S.; NICIURA, S.C.M.; MOLENTO, M.B. Manual Prático: metodologias de diagnóstico da resistência e de detecção de substâncias ativas em parasitas de ruminantes. Embrapa Informação Tecnológica, Brasília, DF, 2011. 153 p.
COLES, G.C. et al. World Association for the Advancement of Veterinary Parasitology (WAAVP) methods for the detection of anthelmintic resistance in nematodes of veterinary importance. Veterinary Parasitology, v. 44, n. 3, p. 35-44, 1992.
COLES, C. G. et al. The detection of anthelmintc resistance in nematodes of veterinary importance. Veterinary Parasitology, v. 136, n. 3, p. 167-185, 2006.
FORTES F.S.; MOLENTO M.B. Resistência anti-helmíntica em nematóides gastrintestinais de pequenos ruminantes: avanços e limitações para seu diagnóstico. Pesquisa Veterinária Brasileira, v.33, n.12, p.1391-1402, 2013.
GORDON, H.M.C.; WHITLOCK, H.V. A new technique for counting nematode eggs in sheep faeces. Journal of Commonwealth Science and Industry Research Organization, v.12, n.1, p.50-52, 1939.
GUGEL, M. et al. Prevalência de endoparasitas em ovinos no oeste de Santa Catarina: resultados preliminares. Synergismus Scyentifica, v.7, n.1, p.1-3, 2012.
KAPLAN, R. M.; VIDYASHANKAR, A. N. An inconvenient truth: Global worming and anthelmintic resistance. Veterinary Parasitology, v.186, n.12, p.70-78, 2012.
ROBERTS, F.H.S.; O´SULLIVAN, J.P. Methods for egg count sand larval cultures for strongyles infesting the gastrointestinal tract of cattle. Autralian Journal of Agricutural Research, v.1, n.1, p.99-102, 1950.
SAMSON-HIMMELSTJERNA, G. et al. Standardization of the egg hatch test for the detection of benzimidazole resistance in parasitic nematodes. Parasitology Research, v.105, n.2, p.825-834, 2009.
SCZESNY-MORAES, E. A. et al. Resistência anti-helmíntica de nematóides gastrintestinais em ovinos, Mato Grosso do Sul. Pesquisa Veterinária Brasileira, v.30, n.3, p.229-236, 2010.
SILVA, M. R. L. et al. Parasitas gastrintestinais de ovinos criados na região de Rondonópolis-MT. Revista Biodiversidade, v.9, n.1, p.67-73, 2010.
UENO, H.; GONÇALVES, P. C. Manual para Diagnóstico das Helmintoses
de Ruminantes. 4ª ed., Tokyo: Japan International Cooperation Agency, 1998. 55p.
WOOD, I.B. et al. World Association for the Advancement of Veterinary Parasitology (W.A.A.V.P.) second edition of guidelines for evaluating the efficacy of anthelmintics in ruminants (bovine, ovine, caprine). Veterinary Parasitology, v.58, n.3, p.181-213, 1995.
In case of approval, the e-mail of acceptance will contain a Letter of Copyright with authors´ name, personal data, and signature of all authors.
