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Validation of diagnostic methods for the detection of Enterospora nucleophila (Microsporidia) in the intestine of gilthead sea bream

AuthorsHossam Eldin, N.; Abu El Ezz, A.; Pozo, R. del; Metwally, A.; Omar, M.; Sitjà-Bobadilla, Ariadna ; Palenzuela, Oswaldo
Issue Date5-Sep-2017
Citation18th International Conference on Diseases of Fish and Shellfish (2017)
AbstractThe microsporidium Enterospora nucleophila is the causative agent of a parasitic emaciative disease in gilthead sea bream (Sparus aurata). This syndrome appears mainly during the winter season in 1. year juvenile gilthead seabream (GSB) as an outstanding growth retardation and size segregation, with serious mortality in some stocks. Diagnosis of E. nucleophila infection is quite challenging due to the minute size of the spores, which develop mainly within the nuclei of enterocytes and rodlet cells in the intestinal epithelium. A remarkable hypercellularity and nuclei alterations in infected cells are commonly observed in the histopathological analysis of infected intestines, but spores are often absent or missed, even in clinical infections. This work describes the optimization and application of a battery of methods for the diagnosis of E. nucleophila infections in GSB. The methods include optimised protocols for fluorescent stain with Calcofluor on tissue sections and smears, a quantitative PCR test, and in-situ hybridization (ISH) techniques. The procedures were applied to a large (>200) panel of clinical cases in order to compare their performance, validate routine qPCR tests, and obtain insights into the pathogenesis of GSB enterosporosis. The work generates validated methods for the diagnosis of E. nucleophila in clinical samples of GSB, which will be valuable for a better understanding of the epidemiology and pathogenesis of this disease. As a follow-up of the analytical and clinical validation covered in this work, a panel of samples will be submitted to diagnostic laboratories participating in an inter-proficiency test as a part of the H2020 project “ParaFishControl”.
DescriptionTrabajo presentado en la 18th International Conference on Diseases of Fish and Shellfish, celebrada en Belfast, del 4 al 8 de septiembre de 2017
Appears in Collections:(IATS) Comunicaciones congresos
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