Development and use of fish cell lines as integrative tools for research in basic science, ecotoxicology, and aquaculture

随着渔业的减少，水产养殖在向全球市场提供鱼类和贝类方面发挥着重要作用。然而，水产养殖的增长受到疾病发生率和高死亡率的限制，这些疾病和死亡率是由于拥挤和污染等紧张的环境条件造成的。水产养殖动物的纯粹性质需要大型水生保持设施，使得具有整个生物体作为模型系统来评估疾病进展的方面和/或评估污染化学品或治疗化合物非常麻烦且非常昂贵。如果感兴趣的病原体的研究是必需的细胞内微生物，则尤其如此。与兼性细胞内病原体或可以在宿主细胞外培养和研究的细胞外病原体相反，专性细胞内病原体难以研究，因为它们需要宿主细胞生长和存活。 随着长期可培养细胞或细胞系的开发和可用性，促进了对病毒（最终的专性细胞内病原体）的研究。对于许多陆生生物，包括人类，来自感兴趣的生物体的细胞培养物已成功地用作研究疾病，药物作用和环境变量在调节其健康中的作用的模型。因此，本提案的目的是研究微孢子虫（一种特征不明确的专性细胞内真菌寄生虫），其致病性，传播，繁殖，宿主特异性和与环境污染物的相互作用的机制，以及开发治疗和预防措施，以控制这些病原体在水产养殖物种使用模式鱼细胞培养物。水产养殖的增长导致越来越多的报告，新兴的机会微孢子虫病原体和微孢子虫病的爆发已成为一个重要的问题，在全球以及加拿大的行业。 Loma salmonae、Loma morhua、Glugea sp.是据报道在加拿大重要鱼类中发生的几种影响鱼类的微孢子虫，本提案将沿着研究可培养微孢子虫模式Anncaliia algerae。

With dwindling fisheries, aquaculture is taking an important role in the provision of fish and shellfish to global markets. However, growth of aquaculture is constrained by incidence of disease and high mortalities due to stressful environmental conditions such as crowding and pollution. The sheer nature of aquacultured animals requiring large aquatic holding facilities, makes it very cumbersome and highly expensive to have whole organisms as model systems to evaluate aspects of disease progression and/or evaluation of contaminating chemicals or of therapeutic compounds. This is especially true if the study of the pathogens of interest, are OBLIGATE intracellular microorganisms. As opposed to facultative intracellular pathogens or to extracellular pathogens that can be cultured and studied outside host cells, obligate intracellular pathogens are difficult to study, as they require host cells for growth and survival. The study of viruses, the ultimate obligate intracellular pathogen, was facilitated with the development and availability of long term culturable cells or cell lines. For many terrestrial organisms, including humans, cell cultures derived from the organisms of interest have been successfully used as models to study disease, drug effects, and the role of environmental variables in modulating their health. Thus the present proposal aims to study MICROSPORIDIA (a poorly characterized obligate intracellular fungal parasites), their mechanisms of pathogenicity, transmission, reproduction, host specificity, and interactions with environmental contaminants as well as to develop therapeutic and preventive measures to control these pathogens in aquacultured species using model fish cell cultures. The growth of aquaculture has led to increasing reports of emerging opportunistic microsporidial pathogens and outbreaks of microsporidiosis have become an important issue in the global as well as Canadian industry. Loma salmonae, Loma morhua, Glugea sp. are some of several fish-afflicting microsporidia that have been reported to occur in important Canadian fish species and will be researched in this proposal along with a model culturable microsporidia, Anncaliia algerae.