Defenses of susceptible and resistant Chinook salmon (Oncorhynchus tshawytscha) against the myxozoan parasite Ceratomyxa shasta.

We investigated intra-specific variation in the response of salmon to infection with the myxozoan Ceratomyxa shasta by comparing the progress of parasite infection and measures of host immune response in susceptible and resistant Chinook salmon Oncorhynchus tshawytscha at days 12, 25 and 90 post exposure. There were no differences in invasion of the gills indicating that resistance does not occur at the site of entry. In the intestine on day 12, infection intensity and Ig(+) cell numbers were higher in susceptible than resistant fish, but histological examination at that timepoint showed more severe inflammation in resistant fish. This suggests a role for the immune response in resistant fish that eliminates some parasites prior to or soon after reaching the intestine. Susceptible fish had a higher IFNγ, IL-6 and IL-10 response at day 12, but all died of fatal enteronecrosis by day 25. The greatest fold change in IFNγ expression was detected at day 25 in resistant Chinook. In addition, the number of Ig(+) cells in resistant Chinook also increased by day 25. By day 90, resistant Chinook had resolved the inflammation, cytokine expression had decreased and Ig(+) cell numbers were similar to uninfected controls. Thus, it appears that the susceptible strain was incapable of containing or eliminating C. shasta but resistant fish: 1) reduced infection intensity during early intestinal infection, 2) elicited an effective inflammatory response in the intestine that eliminated C. shasta, 3) resolved the inflammation and recovered from infection.

Invasion of Ceratomyxa shasta (Myxozoa) and comparison of migration to the intestine between susceptible and resistant fish hosts.

The myxozoan parasite Ceratomyxa shasta infects salmonids causing ceratomyxosis, a disease elicited by proliferation of the parasite in the intestine. This parasite is endemic to the Pacific Northwest of North America and salmon and trout strains from endemic river basins show increased resistance to the parasite. It has been suggested that these resistant fish (i) exclude the parasite at the site of invasion and/or (ii) prevent establishment in the intestine. Using parasites pre-labeled with a fluorescent stain, carboxyfluorescein succinimidyl diacetate (CFSE), the gills were identified as the site of attachment of C. shasta in a susceptible fish strain. In situ hybridization (ISH) of histological sections was then used to describe the invasion of the parasites in the gill filaments. To investigate differences in the progress of infection between resistant and susceptible fish, a C. shasta-susceptible strain of rainbow trout (Oncorhynchus mykiss) and a C. shasta-resistant strain of Chinook salmon (Oncorhynchus tshawytscha) were sampled at consecutive time points following exposure at an endemic site. Using ISH in both species, the parasite was observed to migrate from the gill epithelium into the gill blood vessels where replication and release of parasite stages occurred. Quantitative PCR verified entry of the parasite into the blood. Parasite levels in blood increased 4days p.i. and remained at a consistent level until the second week when parasite abundance increased further and coincided with host mortality. The timing of parasite replication and migration to the intestine were similar for both fish species. The field exposure dose was unexpectedly high and apparently overwhelmed the Chinook salmon's defenses, as no evidence of resistance to parasite penetration into the gills or prevention of parasite establishment in the intestine was observed.

Effects of Ceratomyxa shasta dose on a susceptible strain of rainbow trout and comparatively resistant Chinook and coho salmon.

Ceratomyxa shasta infects salmon and trout, causing ceratomyxosis, a disease characterized by parasite proliferation in the intestine and death. We used laboratory challenges to investigate the infective dose for 3 fish species: a susceptible strain of rainbow trout Oncorhynchus mykiss and comparatively resistant Chinook O. tshawytscha and coho salmon O. kisutch. For susceptible rainbow trout, we determined the outcome of infection under conditions of varying parasite dose, fish size, and parasite concentration. A single actinospore was sufficient to cause a lethal infection in susceptible rainbow trout. The mean days to death (MDD) did not significantly decrease among doses causing 100% prevalence, indicating a minimum time required for parasites to replicate to a fatal level. When dose was constant, but delivered in a higher parasite concentration, higher infection prevalence and mortality resulted. One actinospore fish(-1) caused 57% infection and mortality in fish challenged in 0.5 1 of water, whereas 10 spores fish(-1) resulted in an average of 49% infection and mortality in 1 l challenges. This effect is most likely due to a higher encounter rate in the smaller water volume. Neither infection prevalence nor MDD was significantly different between large trout (84.9 g) and small trout (6.3 g). Chinook salmon did not become infected even when challenged with 5000 actinospores. One fatal infection occurred in coho salmon challenged with 1000 actinospores. This study confirms that even low doses of C. shasta cause severe infection in highly susceptible fish, describes the dose response on MDD, and demonstrates that parasite concentration influences infection prevalence.

Systems level insights into the stress response to UV radiation in the halophilic archaeon Halobacterium NRC-1.

We report a remarkably high UV-radiation resistance in the extremely halophilic archaeon Halobacterium NRC-1 withstanding up to 110 J/m2 with no loss of viability. Gene knockout analysis in two putative photolyase-like genes (phr1 and phr2) implicated only phr2 in photoreactivation. The UV-response was further characterized by analyzing simultaneously, along with gene function and protein interactions inferred through comparative genomics approaches, mRNA changes for all 2400 genes during light and dark repair. In addition to photoreactivation, three other putative repair mechanisms were identified including d(CTAG) methylation-directed mismatch repair, four oxidative damage repair enzymes, and two proteases for eliminating damaged proteins. Moreover, a UV-induced down-regulation of many important metabolic functions was observed during light repair and seems to be a phenomenon shared by all three domains of life. The systems analysis has facilitated the assignment of putative functions to 26 of 33 key proteins in the UV response through sequence-based methods and/or similarities of their predicted three-dimensional structures to known structures in the PDB. Finally, the systems analysis has raised, through the integration of experimentally determined and computationally inferred data, many experimentally testable hypotheses that describe the metabolic and regulatory networks of Halobacterium NRC-1.