Accurate detection of the amphibian fungal pathogen Batrachochytrium dendrobatidis (Bd) is critical for wildlife disease research; however, false negatives in detection do occur. Here we compared different DNA extraction methods to determine the threshold for Bd detection and identify an optimal extraction method to improve detection and quantification of the pathogen. We extracted both lab-created cell suspension standards using PrepMan Ultra, Chelex resin, and 3 spin column DNA extraction kits (Qiagen DNeasy Blood and Tissue, Zymo Quick DNA miniprep, and IBI gMAX mini kit), and further compared extraction methods using field-collected samples. We found that when extracting Bd DNA from cells in lab-created culture, the spin column extraction methods and PrepMan Ultra were equivalent, while the resin method detected higher Bd DNA quantities, especially at higher loads. However, when swabs from live animals were analyzed, low Bd quantities were more than twice as likely to be detected using a spin column extraction than with the PrepMan Ultra extraction method. All tested spin column extraction methods performed similarly across both field and lab samples. Samples containing low Bd quantities yielded inconsistent detection and quantification of Bd DNA copies regardless of extraction method. To manage imperfect detection of Bd, we suggest that presence/absence analyses are more informative than attempting to quantify Bd DNA when quantities are low. Overall, we recommend that a cost-benefit analysis of target species susceptibility and epidemiology be taken into consideration when designing an experiment to determine the most appropriate DNA extraction method to be used, because sometimes detecting low Bd quantities is imperative to the study, whereas in other situations, detecting low DNA quantities is less important.The Atlantic wolffish (AW) and the spotted wolffish (SW) are long-lived fish found in the North Atlantic and Arctic oceans and are respectively classified as special concern and threatened species, mainly due to fisheries bycatch. To better understand health issues associated with the care of these species in public aquaria, reports from all necropsies performed in 2 zoological institutions between 2009 and 2019 were reviewed (31 AW and 8 SW).  https://www.selleckchem.com/mTOR.html  These wolffish were fed with a similar fish-based diet and kept in multi-species exhibits with comparable environmental parameters. The most frequent necropsy findings were the presence of xanthomas (AW 41.9%; SW 75.0%), nephrocalcinosis (AW 42.9%; SW 75.0%) and urocystoliths (AW 6.5%; SW 62.5%). Xanthomas were mostly located at the base of pectoral fins and were characterized by extensive granulomatous inflammation centered on accumulations of partly mineralized degenerate fatty material, mainly composed of cholesterol crystals. Nephrocalcinosis was characterized by the deposition of calcium salts within the renal tubules and was commonly associated with tubular necrosis. The aquarium-housed wolffish were fed a coldwater fish-based diet. However, the natural diet of wolffish is composed mostly of invertebrates such as urchins and crustaceans. Differences in nutrient composition between these diets, such as lipid and mineral content, may have contributed to the development of xanthomatosis, nephrocalcinosis and urocystolithiasis in wolffish housed in these institutions.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 (175 dilution), duration of challenge, lot of fish, and temperature, average mortality in the recirculating system (42%) was lower (p less then 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.Hyperpigmented melanistic skin lesions (HPMLs) of smallmouth bass Micropterus dolomieu are observed in the Potomac and Susquehanna rivers, Chesapeake Bay watershed, USA. Routine, nonlethal population surveys were conducted at 8 sites on the mainstem Susquehanna River and 9 on the Juniata River, a tributary of the Susquehanna River, between 2012 and 2018, and the prevalence of HPMLs was documented. A total of 4078 smallmouth bass were collected from the mainstem Susquehanna River and 6478 from the Juniata River. Lesions were primarily seen in bass greater than 200 mm, and prevalence in the Susquehanna River (8%) was higher (p less then 0.001) than in the Juniata River (2%). As part of ongoing fish health monitoring projects, smallmouth bass were collected at additional sites, primarily tributaries of the Susquehanna (n = 758) and Potomac (n = 545) rivers between 2013 and 2018. Prevalence in the Susquehanna River (13%) was higher (p less then 0.001) than the Potomac (3%). Microscopically, HPMLs were characterized by an increased number of melanocytes in the epidermis or within the dermis and epidermis.