Temporal Dynamics of DNA Methylation Patterns in Response to Rearing Juvenile Steelhead (Oncorhynchus mykiss) in a Hatchery versus Simulated Stream Environment.

Genetic selection is often implicated as the underlying cause of heritable phenotypic differences between hatchery and wild populations of steelhead trout (Oncorhynchus mykiss) that also differ in lifetime fitness. Developmental plasticity, which can also affect fitness, may be mediated by epigenetic mechanisms such as DNA methylation. Our previous study identified significant differences in DNA methylation between adult hatchery- and natural-origin steelhead from the same population that could not be distinguished by DNA sequence variation. In the current study, we tested whether hatchery-rearing conditions can influence patterns of DNA methylation in steelhead with known genetic backgrounds, and assessed the stability of these changes over time. Eyed-embryos from 22 families of Methow River steelhead were split across traditional hatchery tanks or a simulated stream-rearing environment for 8 months, followed by a second year in a common hatchery tank environment. Family assignments were made using a genetic parentage analysis to account for relatedness among individuals. DNA methylation patterns were examined in the liver, a relatively homogeneous organ that regulates metabolic processes and somatic growth, of juveniles at two time points: after eight months of rearing in either a tank or stream environment and after a subsequent year of rearing in a common tank environment. Further, we analyzed DNA methylation in the sperm of mature 2-year-old males from the earlier described treatments to assess the potential of environmentally-induced changes to be passed to offspring. Hepatic DNA methylation changes in response to hatchery versus stream-rearing in yearling fish were substantial, but few persisted after a second year in the tank environment. However, the early rearing environment appeared to affect how fish responded to developmental and environmental signals during the second year since novel DNA methylation differences were identified in the livers of hatchery versus stream-reared fish after a year of common tank rearing. Furthermore, we found profound differences in DNA methylation due to age, irrespective of rearing treatment. This could be due to smoltification associated changes in liver physiology after the second year of rearing. Although few rearing-treatment effects were observed in the sperm methylome, strong family effects were observed. These data suggest limited potential for intergenerational changes, but highlight the importance of understanding the effects of kinship among studied individuals in order to properly analyze and interpret DNA methylation data in natural populations. Our work is the first to study family effects and temporal dynamics of DNA methylation patterns in response to hatchery-rearing.

Relationships between mercury body concentrations, standard metabolic rate, and body mass in eastern mosquitofish (Gambusia holbrooki) from three experimental populations.

Eastern mosquitofish (Gambusia holbrooki) were sampled from three experimental populations (two Hg-exposed populations and one reference population) to determine whether transgenerational exposure (lifelong exposure of multiple generations) to Hg adversely affects standard metabolic rate (SMR). Mosquitofish subjected to lifelong Hg exposure accumulated significant concentrations of Hg in their tissues compared to fish from the reference population (mean: 3.89-4.13 vs 0.08 microHg/g wet mass, respectively). Less than 10% of the variability in Hg tissue concentrations could be explained by fish body mass, likely because of the short life span and/or dietary habits of this species. Despite the high body burdens of Hg in exposed fish, we found no significant difference in SMR among individuals from Hg-exposed or reference populations. Our findings contrast recent laboratory work describing elevated SMR in mosquitofish exposed to 100 microg/L dissolved inorganic Hg for 48 h. To account for contrasting results between studies, we hypothesize that acute exposure to dissolved inorganic Hg damages gill epithelium, resulting in increased metabolic rate, but that lifelong Hg exposure via trophic uptake of methyl mercury does not affect fish respiratory structures. Alternative hypotheses include the possibility that G. holbrooki is a species that can tolerate high body burdens of Hg or that more than four years of genetic isolation during Hg exposure (8-12 generations) resulted in selection for Hg-tolerant or -resistant individuals.

Copper-induced olfactory toxicity in salmon and steelhead: extrapolation across species and rearing environments.

Recent research has shown that hatchery coho salmon (Oncorhynchus kisutch) are vulnerable to the olfactory neurotoxicity caused by copper from urban runoff, pesticide use, and mining activities. To explore the broader application of this data to salmonids living in the wild, we exposed naturally-reared steelhead (O. mykiss) to copper (5 and 20 µg/L; 3h) and measured losses in olfactory function via electro-olfactogram (EOG) recordings. Copper exposure disrupted the olfactory responsiveness of steelhead to an amino acid (L-serine) in a dose-dependent manner that was equivalent to previously published data for hatchery coho. Our findings support extrapolation of copper toxicity data across species and from fish raised in hatcheries to fish in the wild.