Wilson, Chris C

Climate change is a major conservation concern, especially for many cold-adapted species. The rate of warming due to climate change will likely outpace adaptive responses, and many populations will likely need to rely on phenotypic plasticity to cope with environmental warming. It is currently unclear whether plasticity in physiological responses to warming will be sufficient to offset the negative consequences of chronic environmental warming in ectotherms. I studied within-generation and transgenerational plasticity in two cold-adapted species of fishes, lake trout (Salvelinus namaycush) and brook trout (S. fontinalis), following temperature acclimation. Adults of both species were acclimated to either cold or warm temperatures and offspring were generated using a fully factorial breeding design, whereby the family thermal histories included crosses made within each temperature treatment and bidirectional crosses between temperatures. Offspring families were subdivided into two groups and acclimated to either warm or cold temperatures, so that offspring thermal experience matched or mismatched that of one or both parents. Offspring metabolic rate and critical thermal maximum during an acute thermal challenge were measured for both species. Limited transgenerational plasticity was detected in both species, but had a lesser effect than within-generationacclimation. In brook trout, the paternal contribution was greater than the maternal contribution. In lake trout, a mismatch in thermal acclimation, where the offspring were cold-acclimated but the parents warm-acclimated, resulted in elevated offspring metabolic rate without a corresponding increase in growth, suggesting that a mismatch in temperatures across generations could be detrimental to offspring. Using RNA-sequencing, transgenerational plasticity was linked to differential gene expression in the liver of lake trout offspring, in that genes were differentially expressed depending on the parental acclimation temperatures. Within-generation warm acclimation had the greatest effect on gene expression profile of offspring, with more genes differentially expressed under conditions of within-generation warm acclimation compared with transgenerational warm acclimation. Although it has been suggested that transgenerational plasticity may help to buffer the impact of warming due to climate change, my work implies that transgenerational plasticity, like within-generation plasticity, will be insufficient for these two species of cold-adapted salmonids to cope with climate change.

Author Keywords: Brook trout, Climate change, Lake trout, Phenotypic plasticity, Thermal tolerance, Transgenerational plasticity

I investigated the climate vulnerability of a cold-water salmonid by observing the upper thermal tolerance of brook trout (Salvelinus fontinalis) across multiple life stages. Using hatchery- and wild-ancestry fish, I assessed variation in thermal tolerances and carryover effects within and among brook trout life stages and populations that were reared under varying winter thermal regimes as embryos. Embryonic, fry, and gravid adult upper thermal tolerance were tested via CTmax. Warm acclimation was associated with an increase in embryonic upper thermal tolerance. CTmax variability was markedly higher in embryos than juvenile or adult salmonids. Effects of thermal incubation treatments varied by life stage, with incubation temperature and life stage both influencing body size and thermal tolerance. Collectively, these results suggest that brook trout only exhibit short-term carryover effects from thermal stressors during embryo incubation, with no lasting effects on phenotype beyond the first few months (10 weeks) after hatch.

Author Keywords: acclimation response, climate change, ontogeny, salmonid, stress, thermal plasticity

Identifying population declines and mitigating biodiversity loss require reliable monitoring techniques, but complex life histories and cryptic characteristics of anuran species render conventional monitoring challenging and ineffective. Environmental DNA (eDNA) detection is a highly sensitive and minimally invasive alternative to conventional anuran monitoring. In this study, I conducted a field experiment in 30 natural wetlands to compare efficacy of eDNA detection via qPCR to three conventional methods (visual encounter, breeding call, and larval dipnet surveys) for nine anuran species. eDNA and visual encounter surveys detected the greatest species richness, with eDNA methods requiring the fewest sampling events. However, community composition results differed among methods, indicating that even top performing methods missed species detections. Overall, the most effective detection method varied by species, with some species requiring two to three methods to make all possible detections. Further, eDNA detection rates varied by sampling season for two species (A. americanus and H. versicolor), suggesting that species-specific ecology such as breeding and larval periods play an important role in eDNA presence. These findings suggest that optimized monitoring of complex anuran communities may require two or more monitoring methods selected based on the physiology and biology of all target species.

Author Keywords: amphibian, anuran, conventional monitoring, eDNA, environmental DNA, species richness

In the face of climate change, the persistence of cold-adapted species and populations will depend on their capacity for evolutionary adaptation of physiological traits. As a cold-adapted Ice Age relict species, lake trout (Salvelinus namaycush) are ideal for studying potential effects of climate change on coldwater fishes. I studied the thermal acclimation capacity and aerobic metabolism of age 2+ lake trout from four populations across four acclimation temperatures (8ºC, 11ºC, 15ºC, and 19ºC). One population had marginally significant higher active metabolic rate (AMR) and metabolic scope across all temperatures. There was no interpopulation variation for critical thermal maximum (CTM), standard metabolic rate (SMR), or thermal acclimation capacity. Acclimation resulted in a 3ºC increase in thermal tolerance and 3-fold increase in SMR for all populations. At 19ºC, SMR increased and AMR declined, resulting in sharply reduced metabolic scope for all populations. The limited intraspecific variation in thermal physiology suggests that climate change may threaten lake trout at the species rather than population level.

Author Keywords: Climate Change, Lake Trout, Metabolic Rate, Salvelinus namaycush, Temperature, Thermal Acclimation

Researchers have shown increasing interest in biological invasions for the associated ecological and economic impacts as well as for the opportunities they offer to study the mechanisms that induce range expansion in novel environments. I investigated the strategies exhibited by invasive species that facilitate range expansion. Invasive populations exhibit shifts in life-history strategy that may enable appropriate responses to novel biotic and abiotic factors encountered during range expansion. The spatio-temporal scales at which these shifts occur are largely unexplored. Furthermore, it is not known whether the observed dynamic shifts represent a consistent biological response of a given species to range shifts, or whether the shifts are affected by the abiotic characteristics of the new systems. I examined the life-history responses of female round gobies Neogobius melanastomus across fine and coarser spatial scales behind the expansion front and investigated whether invasive populations encountering different environmental conditions (Ontario vs France) exhibited similar life-history shifts. In both study systems, I found an increase in reproductive investment at invasion fronts compared to longer established areas at coarse and fine scales. The results suggest a similar response to range shifts, or a common invasion strategy independent of environmental conditions experienced, and highlight the dynamic nature of an invasive population's life history behind the invasion front.

The second part of my research focused on the development of an appropriate eDNA method for detecting invasive species at early stages of invasion to enable early detection and rapid management response. I developed a simple, inexpensive device for collecting water samples at selected depths for eDNA analysis, including near the substrate where eDNA concentration of benthic species is likely elevated. I also developed a protocol to optimise DNA extraction from water samples that contain elevated concentration of inhibiters, in particular near-bottom samples. Paired testing of eDNA and conventional surveys was used to monitor round goby expansion along its invasion pathway. Round gobies were detected in more sites with eDNA, permitting earlier, more accurate, upstream detection of the expansion front. My study demonstrated the accuracy and the power of using eDNA survey method to locate invasion fronts.

Author Keywords: Age-specific reproductive investment, DNA extraction, Energy allocation, Fecundity, Invasion front, Range expansion

Effective population size (Ne) is a foundational concept in conservation biology, in part due to its relationship to the adaptive potential of populations. Although Ne is often estimated for wild populations, it is rarely calibrated against actual population estimates (Nc) other than to produce Ne/Nc ratios. This project used demographic and genetic data for from two intensively-studied populations of lake trout (Salvelinus namaycush) in Ontario's Experimental Lake Area (ELA) as baseline data for evaluating the performance of multiple Ne estimators. Several temporal and single-time (point) genetic methods of estimating Ne were compared against demographic Ne estimates and known population data, as well as variation and consistency within and among Ne estimators. Changes in genetic Ne estimates over time were also compared to changes in demographic structure and fluctuating census estimates, including the effect of an experimentally manipulated population bottleneck on demographic and genetic Ne estimates during population reduction and recovery. Sampling years that included the most pre-, during and post-bottleneck data revealed the lowest estimates using temporal estimators (Ne = 16 to 18) despite pre- and post-bottleneck census estimates of 591 and 565. Estimation of Ne had increasingly tighter confidence intervals as sample sizes approached the actual number of breeding individuals in each population. Performance differences among the tested estimators highlight their potential biases and reliance on different assumptions, illustrating their potential value and caveats for assessing adaptive potential of wild populations.

Author Keywords: Effective Population Size, Experimental Lakes Area, Fish Population Assessment, Lake Trout, Population Demographics, Population Genetics

Understanding the maintenance and spread of invasive diseases is critical in evaluating threats to biodiversity and how to best minimize their impact, which can by done by monitoring disease occurrences across time and space. I sought to apply existing and upcoming molecular tools to assess fluctuations in both presence and strain variation of frog virus 3 (FV3), a species of Ranavirus, across Canadian waterbodies. I explored the temporal patterns and spatial distribution of ranavirus presence across multiple months and seasons using environmental DNA techniques. Results indicate that ranavirus was present in approximately 72.5% of waterbodies sampled on a fine geographical scale (<10km between sites, 7,150 km2), with higher detection rates in later summer months than earlier. I then explored the sequence variability at the major capsid protein gene (MCP) and putative virulence gene (vIF-2α) of FV3 samples from Ontario, Alberta, and the Northwest Territories, with the premise of understanding pathogen movement across the landscape. However, a lack of genetic diversity was found across regions, likely due to a lack of informative variation at the chosen genetic markers or lack of mutation. Instead, I found a novel FV3-like ranavirus and evidence for a recombinant between FV3 and a ranavirus of another lineage. This thesis provides a deeper understanding into the spatio-temporal distribution of FV3, with an idea of how widespread and threatening ranaviruses are to amphibian diversity.

Keywords: ranavirus, frog virus 3, amphibians, environmental DNA, phylogenetics, wildlife disease, disease surveillance, major capsid protein, vIF-2α

Author Keywords: amphibians, environmental DNA, frog virus 3, phylogenetics, ranavirus, wildlife disease

North American freshwater fishes are declining rapidly due to habitat fragmentation, degradation, and loss. In some cases, translocations can be used to reverse local extirpations by releasing species in suitable habitats that are no longer naturally accessible. Lake sturgeon (Acipenser fulvescens) experienced historical overharvest across their distribution, leading to endangered species listings and subsequent protection and recovery efforts. Despite harvest and habitat protections, many populations do not appear to be recovering, which has been attributed to habitat alteration and fragmentation by dams. In 2002, 51 adult lake sturgeon from the Mattagami River, Ontario, Canada were translocated 340 km upstream to a fragmented 35 km stretch of the river between two hydroelectric generating stations, where sturgeon were considered extirpated. This study assessed the translocation effort using telemetry (movement), demographics and genetic data. Within the first year, a portion of the radio-tagged translocated individuals dispersed out of the release area, and released radio-tagged individuals used different areas than individuals radio-tagged ten years later. Catches of juvenile lake sturgeon have increased over time, with 150 juveniles caught within the duration of this study. The reintroduced population had similar genetic diversity as the source population, with a marked reduction in effective population size (Ne). The results indicate that the reintroduction effort was successful, with evidence of successful spawning and the presence of juvenile lake sturgeon within the reintroduction site. Overall, the results suggest adult translocations may be a useful tool for re-establishing other extirpated lake sturgeon populations.

Author Keywords: conservation, endangered species, lake sturgeon, reintroduction, telemetry, translocation

Environmental DNA (eDNA) detection uses species-specific markers to screen DNA from bulk samples, such as water, to infer species presence. This study involved the development and testing of species-specific markers for four freshwater pearly mussels (Unionidae). The markers were applied to water samples from intensively sampled mussel monitoring sites to compare species detections from eDNA with established sampling method detections. Target species were detected using eDNA at all sites where they had previously been detected by quadrat sampling. This paired design demonstrated that eDNA detection was at least as sensitive as quadrat sampling and that high species specificity can be achieved even when designing against many sympatric unionids. Detection failures can impede species conservation efforts and occupancy estimates; eDNA sampling could improve our knowledge of species distributions and site occupancy through increased sampling sensitivity and coverage.

Author Keywords: conservation genetics, cytochrome oxidase subunit I (COI), environmental DNA (eDNA), quantitative PCR (qPCR), species at risk (SAR)

Recent range reductions of endangered species have been linked to urban development, increased agricultural activities, and introduction of non-native species. I used Redside Dace (Clinostomus elongatus) as a focal species to examine the utility of novel monitoring approaches, and to understand historical and contemporary processes that have influenced their present distribution. I tested the efficacy of environmental DNA (eDNA) to detect Redside Dace, and showed that eDNA was more sensitive for detecting species presence than traditional electrofishing. Parameters such as season, number of replicates, and spatial versus temporal sampling need to be accounted for when designing an eDNA monitoring program, as they influence detection effectiveness and power. I also assessed the species' phylogeographic structure using both mitochondrial and microsatellite DNA analysis. The data from the microsatellite markers indicate that Redside Dace populations are genetically structured, with the exception of several populations from the Allegheny River basin. Combined sequence data from three mitochondrial genes (cytochrome b, ATPase 6 and ATPase 8) indicated that Redside Dace persisted within three Mississippian refugia during the last glaciation. Secondary contact between two lineages was indicated by both mitochondrial and microsatellite data. The combined results from the eDNA and conservation genetics studies can be used to inform Redside Dace recovery efforts, and provide a template for similar efforts for other aquatic endangered species.

Author Keywords: eDNA, endangered, genetics, phylogeography