| Summary: | Poster presentado en el 15th International Conference on Diseases of Fish and Shellfish celebrado en Croacia del 12 al 16 de septiembre de 2011 Enteromyxum scophthalmi and E. leei are myxozoan parasites producing severe diseases in marine aquaculture. These intestinal parasites cause catarrhal enteritis and emaciative syndromes and they have direct, fish-to-fish transmission, and wide host spectrum. E. scophthalmi infection typically results in 100% loses in turbot stocks, and E. leei produces chronic losses in sea bream culture by a rather slow-progressing enteritis. Due to the lack of efficacious treatments only management measures such as hygiene and culling of infected stocks are currently available. Identification of relevant Enteromyxum genes, expressed in the course of their development in the fish, would provide essential information related to the mechanisms of invasion, proliferation, maturation and virulence. These data would offer new opportunities to interfere in the parasites development and to design strategies for enteromyxoses control. Next-generation sequencing techniques are able to generate large amounts of genetic data and constitute an ideal platform for gene discovery in myxozoan parasites. The objective of the current work was to develop methods for parasite purification for downstream analysis and identification of expressed genes. Enteromyxum spp. are histozoic parasites developing in close contact with host cells within the intestinal mucosa and cannot be grown in vitro. Previous attempts to isolate developmental stages by mechanical and chemical techniques have been unsuccessful. A method was developed to generate cell suspensions containing free parasites from Enteromyxum-infected intestines by adapting protocols for purification of GALT lymphocytes [1, 2]. Briefly, pieces of intestine were cut and the mucosal layer was detached and cells disaggregated in buffer containing DTT and EDTA. The resulting cell suspension was washed and filtered through cell strainers (40 µm) and cell ...
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