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Disease epidemiology requires information about ecological and environmental condi- tions to identify factors that can influence disease progression. Bacterial kidney disease (BKD) is an endemic disease among Pacific Northwest salmonids that causes significant morbidity and mortality in artificially propagated stocks, but risk factors for infection among free-living salmon are unknown. We evaluated infection by the causative agent of BKD, Renibacterium salmoninarum, in 1752 fish across 52 sampling sites monthly from May to November 2003 as a component of a broader study of neritic habitat use in Puget Sound by juvenile Chinook salmon Oncorhynchus tshawytscha. Infection intensity was ≤10 cells per slide for 77% of the fish. Correlations between the density of Chinook salmon with infection prevalence and with infection intensity were observed across multiple spatial scales. Capture location was a stronger predictor of infection than fish origin (based on coded wire tags) or genetic stock. Influential risk factors by logistic regression were temperature, densities of marked and unmarked Chinook salmon, and density of river lamprey Lampetra ayresis. Renibac- terium salmoninarum were found in gut contents and kidney of river lamprey, suggesting this species may be a transmission vector. The low infection intensity, lack of an effect of fish origin, effect of capture bay, and strong associations with Chinook salmon density are consistent with horizontal transmission of R. salmoninarum during the juvenile neritic phase, posing a potential for infectious interaction between sympatric hatchery and wild fish.

Renibacterium salmoninarum causes bacterial kidney disease (BKD), a chronic and sometimes fatal disease of salmon and trout that could lower fitness in populations with high prevalences of infection. Prevalence of R. salmoninarum infection among juvenile Chinook salmon Oncorhynchus tshawytscha inhabiting neritic marine habitats in North Puget Sound, Washington, USA, was assessed in 2002 and 2003. Fish were collected by monthly surface trawl at 32 sites within 4 bays, and kidney infections were detected by a quantitative fluorescent antibody technique (qFAT). The sensitivity of the qFAT was within an order of magnitude of the quantitative real-time PCR (qPCR) sensitivity. Prevalence of infection was classified by fish origin (marked/hatchery vs. unmarked/likely natural spawn), month of capture, capture location and stock origin. The highest percentages of infected fish (63.5 to 63.8%) and the greatest infection severity were observed for fish collected in Bellingham Bay. The lowest percentages were found in Skagit Bay (11.4 to 13.5%); however, there was no difference in prevalence between marked and unmarked fish among the capture locations. The optimal logistic regression model of infection probabilities identified the capture location of Bellingham Bay as the strongest effect, and analysis of coded wire tagged (CWT) fish revealed that prevalence of infection was associated with the capture location and not with the originating stock. These results suggest that infections can occur during the early marine life stages of Chinook salmon that may be due to common reservoirs of infection or horizontal transmission among fish stocks.

Although there are a variety of methods available for the detection of Renibacterium salmoninarum, the causative agent of bacterial kidney disease in salmon and trout, the enzyme-linked immunosorbent assay (ELISA) is probably the most widely used method. However, ELISA measures bacterial antigen, which does not necessarily reflect the number of cells present. We hypothesized that dual analysis of kidney tissue by ELISA and a quantitative real-time polymerase chain reaction assay (qPCR) would provide complementary information about antigen level and the number of bacterial genomes. We found that DNA extracted from the insoluble fraction of the ELISA tissue preparation produced the same qPCR result as DNA extracted directly from frozen tissue, permitting true dual analysis of the same tissue sample. We examined kidney tissue in this manner from individual free-ranging juvenile Chinook salmon and antibiotic-treated captive subadult Chinook salmon and observed 3 different patterns of results. Among the majority of fish, there was a strong correlation between the ELISA value and the qPCR value. However, subsets of fish exhibited either low ELISA values with elevated qPCR values or higher ELISA values with very low qPCR values. These observations suggest a conceptual model that allows inferences about the state of infection of individual fish based on dual ELISA/qPCR results. Although this model requires further assessment through experimental infections and treatments, it may have utility in broodstock selection programs that currently apply egg-culling practices based on ELISA alone.