Since their first discovery in the 19th century, microsporidian species have been found to be successful obligate intracellular parasites capable of infecting a wide variety of hosts including economically and ecologically important organisms as well as model organisms for biomedical research. Recently, cases of infection of the widely used animal model Danio rerio, commonly known as the zebrafish, by the newly described microsporidium Pseudoloma neurophilia have been reported in an increasing number of research facilities. Current knowledge of the biology of this parasite found in 75% of the Zebrafish Resource Center facilities is limited to microscopic analyses on its lifecycle as well as its physical, behavioral and psychic impact on its hosts. Despite the growing relevance of this parasite in biomedical research no current data is available on its genome. In this dissertation, I provide additional knowledge on the basic biology of P. neurophilia by acquiring and exploring the content and structure of the first genome draft of the zebrafish parasite. My findings reveal that the 5.25 Mb genome of P. neurophilia harbors an unusually high amount of transposable elements as well as numerous inserts found in coding regions typically conserved in microsporidia and other organisms. This peculiar obligate parasite demonstrates strong phylogenetic and genetic relationships with other fish-mosquito microsporidia. Similar to what is observed in closely related species, intra-genomic analyses of P. neurophilia’s genome suggest that it is diploid and possesses a large repertoire of over a thousand putative genes unique to this specie. Overall, my findings provide new insights into the basic biology of this parasite and represent a milestone in the understanding of P. neurophilia and D. rerio host-parasite interaction and ultimately in the development of treatments against this parasite that has been infecting the zebrafish research industry for the past decades.

Zebrafish are becoming increasingly popular to use in different kinds of research projects as research animals, replacing rodents in many fields. When using animals for research, it is important to keep track of the animal health in order to get reliable results. The purpose of the study was to investigate whether these pathogens could be found analyzing environmental materials with real-time PCR instead of euthanizing fish and submitting them for histopathology. Also, to see if any material differentiated from the rest regarding accessibility to work with in a routine diagnostic laboratory.     This study was performed on environmental samples, such as filters, swabs, detritus and water, from a recirculating water system holding zebrafish. The pathogens analyzed were Mycobacterium chelonae, M. haemophilum, M. abscessus, M. marinum, M. fortuitum and Pseudoloma neurophilia, all common pathogens that can affect zebrafish.     All materials tested gave at least one positive result for most of the pathogens tested. Two pathogens were not detected, M. marinum and M. abscessus. Due to poorly working PCR-system for M. fortuitum, the results for that bacteria were deemed inconclusive. The filter materials and the swabs of the filter materials gave the best results in this small study, although all materials gave satisfactory results. In conclusion this study shows that environmental samples can be used to detect pathogens in zebrafish, but larger studies should be performed to better evaluate which material is the best one to use.