Flavobacterium columnare immersion challenges are affected by water-related environmental parameters and thus are difficult to reproduce. Whereas these challenges are typically conducted using flow-through systems, use of a recirculating challenge system to control environmental parameters may improve reproducibility. We compared mortality, bacterial concentration, and environmental parameters between flow-through and recirculating immersion challenge systems under laboratory conditions using 20 rainbow trout families. Despite identical dose concentration (1:75 dilution), duration of challenge, lot of fish, and temperature, average mortality in the recirculating system (42%) was lower (p < 0.01) compared to the flow-through system (77%), and there was low correlation (r = 0.24) of family mortality. Mean days to death (3.25 vs. 2.99 d) and aquaria-to-aquaria variation (9.6 vs. 10.4%) in the recirculating and flow-through systems, respectively, did not differ (p ≥ 0.30). Despite 10-fold lower water replacement rate in the recirculating (0.4 exchanges h-1) compared to flow-through system (4 exchanges h-1), differences in bacterial concentration between the 2 systems were modest (≤0.6 orders of magnitude) and inconsistent throughout the 21 d challenge. Compared to the flow-through system, dissolved oxygen during the 1 h exposure and pH were greater (p ≤ 0.02), and calcium and hardness were lower (p ≤ 0.03), in the recirculating system. Although this study was not designed to test effects of specific environmental parameters on mortality, it demonstrates that the cumulative effects of these parameters result in poor reproducibility. A recirculating immersion challenge model may be warranted to empirically identify and control environmental parameters affecting mortality and thus may serve as a more repeatable laboratory challenge model.
β‐zeolite assisted lignin‐first fractionation in a flow‐through reactor