The Little Bighorn River is the primary source of water for water treatment plants serving the local Crow Agency population, and has special significance in the spiritual and ceremonial life of the Crow tribe. Unfortunately, the watershed suffers from impaired water quality, with high counts of fecal coliform bacteria routinely measured during run-off events. A metagenomic analysis was carried out to identify potential pathogens in the river water. The Oxford Nanopore MinION platform was used to sequence DNA in near real time to identify both uncultured and a coliform-enriched culture of microbes collected from a popular summer swimming area of the Little Bighorn River. Sequences were analyzed using CosmosID bioinformatics and, in agreement with previous studies, enterohemorrhagic and enteropathogenic Escherichia coli and other E. coli pathotypes were identified. Noteworthy was detection and identification of enteroaggregative E. coli O104:H4 and Vibrio cholerae serotype O1 El Tor, however, cholera toxin genes were not identified. Other pathogenic microbes, as well as virulence genes and antimicrobial resistance markers, were also identified and characterized by metagenomic analyses. It is concluded that metagenomics provides a useful and potentially routine tool for identifying in an in-depth manner microbial contamination of waterways and, thereby, protecting public health.

Riverine ecosystems are biogeochemical powerhouses driven largely by microbial communities that inhabit water columns and sediments. Because rivers are used extensively for anthropogenic purposes (drinking water, recreation, agriculture, and industry), it is essential to understand how these activities affect the composition of river microbial consortia. Recent studies have shown that river metagenomes vary considerably, suggesting that microbial community data should be included in broad-scale river ecosystem models. But such ecogenomic studies have not been applied on a broad “aquascape” scale, and few if any have applied the newest nanopore technology. We investigated the metagenomes of 11 rivers across 3 continents using MinION nanopore sequencing, a portable platform that could be useful for future global river monitoring. Up to 10 Gb of data per run were generated with average read lengths of 3.4 kb. Diversity and diagnosis of river function potential was accomplished with 0.5–1.0 ⋅ 10⁶ long reads. Our observations for 7 of the 11 rivers conformed to other river-omic findings, and we exposed previously unrecognized microbial biodiversity in the other 4 rivers. Deeper understanding that emerged is that river microbial consortia and the ecological functions they fulfil did not align with geographic location but instead implicated ecological responses of microbes to urban and other anthropogenic effects, and that changes in taxa manifested over a very short geographic space.

Neonicotinoids (NEO) represent the main class of insecticides currently in use, with thiamethoxam (THX) and clothianidin (CLO) primarily applied agriculturally. With few comprehensive studies having been performed with non-target amphibians, the aim was to investigate potential biomarker responses along an adverse outcome pathway of NEO exposure, whereby data were collected on multiple biological hierarchies. Juvenile African clawed frogs, Xenopus laevis, were exposed to commercial formulations of THX and CLO at high (100 ppm) and low (20 ppm) concentrations of the active ingredient. Mortality, growth, development, liver metabolic enzyme activity, and gene expression endpoints were quantified. Tadpoles (n > 1000) from NF 47 through tail resorption stage (NF 66) were exposed to NEO or to NEO-free media treatments. Liver cell reductase activity and cytotoxicity were quantified by flow cytometry. Compared to control reference gene expressions, levels of expression for NEO receptor subunits, cell structure, function, and decontamination processes were measured by RT-qPCR by using liver and brain. Mortality in THX high was 21.5% compared to the control (9.1%); the metabolic conversion of THX to CLO may explain these results. The NF 57 control tadpoles were heavier, longer, and more developed than the others. The progression of development from NF 57–66 was reduced by THX low, and weight gain was impaired. Liver reductases were highest in the control (84.1%), with low NEO exhibiting the greatest reductions; the greatest cytotoxicity was seen with THX high. More transcriptional activity was noted in brains than in livers. Results affirm the utility of a study approach that considers multiple complexities in ecotoxicological studies with non-target amphibians, underscoring the need for simultaneously considering NEO concentration-response relationships with both whole-organism and biomarker endpoints.

Walleyes Sander vitreus support important recreational and commercial fisheries, and have been introduced into many systems throughout the United States and Canada. Use of sterile triploid fish is a valuable management strategy for protecting native fish species from naturally reproducing populations of introduced predators like Walleyes. Hydrostatic pressure is one of the more effective methods for inducing triploidy in Walleyes, and the technique is currently evolving. A 2.7‐L capacity electric pressure chamber (TRC‐APV Aqua Pressure Vessel; TRC Hydraulics) was used to produce triploid Walleyes using hydrostatic pressure with eggs collected from Pueblo Reservoir, Colorado. The TRC‐APV has many advantages over manually operated pressure chambers including its portability, large capacity, and safety. Here, the methods used to create triploid Walleyes using the TRC‐APV and the flow cytometric methods for estimating the efficacy of ploidy induction from pooled samples of fry are described. In addition, times of initiation (TIs), the times between fertilization and when eggs were subjected to pressure, were varied to determine the optimal time for maximizing induction and hatching success rates. Results suggest that triploidy in Walleyes is maximized with a TI of approximately 7.5 min, whereas hatching success is maximized with a TI of just over 8 min.

From 24 mating sets, 6300 fingerling of yellow perch (Perca flavescens) were stocked into one pond and equal numbers of progeny from six representative sets out of the 24 were stocked into each of two other ponds. After communal rearing for 21 months, total length and body weight were assessed for n=300 fish in each of the three ponds and molecular pedigrees were performed for each sampled individual to assign the progeny back to the original parents. The overall average number of alleles per locus was A=16.4 and observed and expected heterozygosities were H_o=0.88 and H_e=0.77 respectively. The mean weight of random samples and the top 10% fast-growing fish from the pond with all the sets was significantly greater than those from either of the two replicate ponds with six crosses. For the two replicate ponds, no significant differences were found in family rankings and assignment of the top 10% fast-growing fish, indicating that families with superior growth performance in one pond also exhibited the same superior growth performance in the replicate pond. However, there were no significant correlations detected in family mean weights of the top 10% fish between any two of the three ponds.