Thermal stressors during embryo incubation have limited ontogenic carryover effects in brook trout.

Winter climate is changing rapidly in northern latitudes, and these temperature events have effects on salmonid thermal biology. Stressors during winter egg incubation could reduce hatching success and physiological performance of fall-spawning fishes. Here we quantified the potential for ontogenic carryover effects from embryonic thermal stress in multiple wild and hatchery-origin populations of brook trout (Salvelinus fontinalis), a temperate ectotherm native to northeastern North America. Fertilized eggs from four populations were incubated over the winter in the laboratory in four differing thermal regimes: ambient stream-fed water, chronic warming (+2 °C), ambient with a mid-winter cold-shock, and short-term warming late during embryogenesis (to stimulate an early spring). We examined body size and upper thermal tolerance at the embryonic, fry (10 weeks post-hatch and 27-30 weeks post-hatch) and gravid adult (age 2+) life stages (overall N = 1482). In a separate experiment, we exposed developing embryos to acute seven-day heat stress events immediately following fertilization and at the eyed-egg stage, and then assessed upper thermal tolerance (CTmax) 37 weeks post-hatch. In all cases, fish were raised in common garden conditions after hatch (i.e., same temperatures). Our thermal treatments during incubation had effects that varied by life stage, with incubation temperature and life stage both affecting body size and thermal tolerance. Embryos incubated in warmer treatment groups had higher thermal tolerance; there was no effect of the mid-winter melt event on embryo CTmax. Ten weeks after hatch, fry from the ambient and cold-shock treatment groups had higher and less variable thermal tolerance than did the warmer treatment groups. At 27-30 post-hatch and beyond, differences in thermal tolerance among treatment groups were negligible. Collectively, our study suggests 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 after hatch.

CTmax in brook trout (Salvelinus fontinalis) embryos shows an acclimation response to developmental temperatures but is more variable than in later life stages.

Critical thermal maximum (CTmax ) is widely used to measure upper thermal tolerance in fish but is rarely examined in embryos. Upper thermal limits generally depend on an individual's thermal history, which molds plasticity. We examined how thermal acclimation affects thermal tolerance of brook trout (Salvelinus fontinalis) embryos using a novel method to assess CTmax in embryos incubated under three thermal regimes. Warm acclimation was associated with an increase in embryonic upper thermal tolerance. However, CTmax variability was markedly higher than is typical for juvenile or adult salmonids.

Optimization of an amplicon sequencing-based microsatellite panel and protocol for stock identification and kinship inference of lake trout (Salvelinus namaycush).

Kinship-based methods of population assessment such as close-kin mark-recapture require accurate and efficient genotyping methods capable of resolving complex relationships among kin. Inference of such relationships can be difficult using biallelic loci due to the large number of markers required to obtain the necessary power. Sequencing-based microsatellite panels offer an efficient alternative, combining high polymorphism with efficient next-generation methods. Here we construct, optimize, and test one such panel for lake trout (Salvelinus namaycush) using a combination of previously-published loci adapted for sequencing and de novo loci mined from a genome assembly. We performed three rounds of primer optimization, yielding a final panel of 131 loci, followed by testing with two different levels of PCR multiplexing (all primers in one or two groups) and two different reaction volumes (5 and 10 µL). Our results showed that the use of the largest multiplex and smallest reaction volume did not substantially change results, allowing significant cost and time savings. To test panel accuracy, we used both a set of 153 known-origin samples from origins of management interest and a series of hatchery crosses representing nine families with parent-offspring, half-sibling, and largely-unrelated pairs. Our results indicate that sequencing-based microsatellite panels can efficiently and accurately provide the information required for a population genetics analyses including population assignment, calculation of between-population F ST, and kinship-based population estimation techniques. Such techniques are seeing increasing applications for a wide range of taxa; our findings should provide insight and guidance for the development of the necessary molecular resources.

Short-term acclimation dynamics in a coldwater fish.

Critical thermal maximum (CTmax) is widely used for measuring thermal tolerance but the strong effect of acclimation on CTmax is a likely source of variation within and among studies/species that makes comparisons more difficult. There have been surprisingly few studies focused on quantifying how quickly acclimation occurs or that combine temperature and duration effects. We studied the effects of absolute temperature difference and duration of acclimation on CTmax of brook trout (Salvelinus fontinalis), a well-studied species in the thermal biology literature, under laboratory conditions to determine how each of the two factors and their combined effects influence critical thermal maximum. Using an ecologically-relevant range of temperatures and testing CTmax multiple times between one and 30 days, we found that both temperature and duration of acclimation had strong effects on CTmax. As predicted, fish that were exposed to warmer temperatures longer had increased CTmax, but full acclimation (i.e., a plateau in CTmax) did not occur by day 30. Therefore, our study provides useful context for thermal biologists by demonstrating that the CTmax of fish can continue to acclimate to a new temperature for at least 30 days. We recommend that this be considered in future studies measuring thermal tolerance that intend to have their organisms fully acclimated to a given temperature. Our results also support using detailed thermal acclimation information to reduce uncertainty caused by local or seasonal acclimation effects and to improve the use of CTmax data for fundamental research and conservation planning.

A new GTSeq resource to facilitate multijurisdictional research and management of walleye Sander vitreus.

Conservation and management professionals often work across jurisdictional boundaries to identify broad ecological patterns. These collaborations help to protect populations whose distributions span political borders. One common limitation to multijurisdictional collaboration is consistency in data recording and reporting. This limitation can impact genetic research, which relies on data about specific markers in an organism's genome. Incomplete overlap of markers between separate studies can prevent direct comparisons of results. Standardized marker panels can reduce the impact of this issue and provide a common starting place for new research. Genotyping-in-thousands (GTSeq) is one approach used to create standardized marker panels for nonmodel organisms. Here, we describe the development, optimization, and early assessments of a new GTSeq panel for use with walleye (Sander vitreus) from the Great Lakes region of North America. High genome-coverage sequencing conducted using RAD capture provided genotypes for thousands of single nucleotide polymorphisms (SNPs). From these markers, SNP and microhaplotype markers were chosen, which were informative for genetic stock identification (GSI) and kinship analysis. The final GTSeq panel contained 500 markers, including 197 microhaplotypes and 303 SNPs. Leave-one-out GSI simulations indicated that GSI accuracy should be greater than 80% in most jurisdictions. The false-positive rates of parent-offspring and full-sibling kinship identification were found to be low. Finally, genotypes could be consistently scored among separate sequencing runs >94% of the time. Results indicate that the GTSeq panel that we developed should perform well for multijurisdictional walleye research throughout the Great Lakes region.

Within-Generation and Transgenerational Plasticity of a Temperate Salmonid in Response to Thermal Acclimation and Acute Temperature Stress.

AbstractThe rise in temperature associated with climate change may threaten the persistence of stenothermal organisms with limited capacities for beneficial thermal acclimation. We investigated the capacity for within-generation and transgenerational thermal responses in brook trout (Salvelinus fontinalis), a cold-adapted salmonid. Adult fish were acclimated to temperatures within (10°C) and above (21°C) their thermal optimum for 6 mo before spawning, then mated in a full factorial breeding design to produce offspring from cold- and warm-acclimated parents and bidirectional crosses between parents from both temperature treatments. Offspring from families were subdivided and reared at two acclimation temperatures representing their current (15°C) and anticipated future (19°C) habitat temperatures. Offspring thermal physiology was measured as the rate of oxygen consumption (Mo2) during an acute change in temperature (increase of 2°C h-1) to observe their Mo2-temperature relationship. We recorded resting Mo2, peak (highest achieved, thermally induced) Mo2, and critical thermal maximum (CTM) as performance metrics. Although limited, within-generation plasticity was greater than transgenerational plasticity, with offspring warm acclimation elevating CTM by 0.5°C but slightly lowering peak thermally induced Mo2. Transgenerational plasticity was evident as a slightly elevated resting Mo2 and a shift of the Mo2-temperature relationship to higher rates overall in offspring from warm-acclimated parents. Furthermore, offspring whose parents were warm acclimated were in worse condition than those whose parents were cold acclimated. Both parents contributed to offspring thermal responses; however, the paternal effect was stronger. Despite the existence of within-generation and transgenerational plasticity in brook trout, it is unlikely that these will be sufficient for coping with long-term changes to environmental temperatures.

A chromosome-anchored genome assembly for Lake Trout (Salvelinus namaycush).

Here, we present an annotated, chromosome-anchored, genome assembly for Lake Trout (Salvelinus namaycush) - a highly diverse salmonid species of notable conservation concern and an excellent model for research on adaptation and speciation. We leveraged Pacific Biosciences long-read sequencing, paired-end Illumina sequencing, proximity ligation (Hi-C) sequencing, and a previously published linkage map to produce a highly contiguous assembly composed of 7378 contigs (contig N50 = 1.8 Mb) assigned to 4120 scaffolds (scaffold N50 = 44.975 Mb). Long read sequencing data were generated using DNA from a female double haploid individual. 84.7% of the genome was assigned to 42 chromosome-sized scaffolds and 93.2% of Benchmarking Universal Single Copy Orthologues were recovered, putting this assembly on par with the best currently available salmonid genomes. Estimates of genome size based on k-mer frequency analysis were highly similar to the total size of the finished genome, suggesting that the entirety of the genome was recovered. A mitochondrial genome assembly was also produced. Self-versus-self synteny analysis allowed us to identify homeologs resulting from the salmonid specific autotetraploid event (Ss4R) as well as regions exhibiting delayed rediploidization. Alignment with three other salmonid genomes and the Northern Pike (Esox lucius) genome also allowed us to identify homologous chromosomes in related taxa. We also generated multiple resources useful for future genomic research on Lake Trout, including a repeat library and a sex-averaged recombination map. A novel RNA sequencing data set for liver tissue was also generated in order to produce a publicly available set of annotations for 49,668 genes and pseudogenes. Potential applications of these resources to population genetics and the conservation of native populations are discussed.

Limited transgenerational effects of environmental temperatures on thermal performance of a cold-adapted salmonid.

The capacity of ectotherms to cope with rising temperatures associated with climate change is a significant conservation concern as the rate of warming is likely too rapid to allow for adaptative responses in many populations. Transgenerational plasticity (TGP), if present, could potentially buffer some of the negative impacts of warming on future generations. We examined TGP in lake trout to assess their inter-generational potential to cope with anticipated warming. We acclimated adult lake trout to cold (10°C) or warm (17°C) temperatures for several months, then bred them to produce offspring from parents within a temperature treatment (cold-acclimated and warm-acclimated parents) and between temperature treatments (i.e. reciprocal crosses). At the fry stage, offspring were also acclimated to cold (11°C) or warm (15°C) temperatures. Thermal performance was assessed by measuring their critical thermal maximum (CTM) and the change in metabolic rate during an acute temperature challenge. From this dataset, we also determined their resting and peak (highest achieved, thermally induced) metabolic rates. There was little variation in offspring CTM or peak metabolic rate, although cold-acclimated offspring from warm-acclimated parents exhibited elevated resting metabolic rates without a corresponding increase in mass or condition factor, suggesting that transgenerational effects can be detrimental when parent and offspring environments mismatch. These results suggest that the limited TGP in thermal performance of lake trout is unlikely to significantly influence population responses to projected increases in environmental temperatures.

Mixed-stock analysis using Rapture genotyping to evaluate stock-specific exploitation of a walleye population despite weak genetic structure.

Mixed-stock analyses using genetic markers have informed fisheries management in cases where strong genetic differentiation occurs among local spawning populations, yet many fisheries are supported by multiple, weakly differentiated stocks. Freshwater fisheries exemplify this problem, with many populations supported by multiple stocks of young evolutionary age and isolated across small spatial scales. Consequently, attempts to conduct genetic mixed-stock analyses of inland fisheries have often been unsuccessful. Advances in genomic sequencing offer the ability to discriminate among populations with weak population structure, providing the necessary resolution to conduct mixed-stock assignment among previously indistinguishable stocks. We used genomic data to conduct a mixed-stock analysis of eastern Lake Erie's commercial and recreational walleye (Sander vitreus) fisheries and estimate the relative harvest of weakly differentiated stocks (pairwise F ST < 0.01). Using RAD-capture (Rapture), we sequenced and genotyped individuals from western and eastern basin local spawning stocks at 12,081 loci with 95% reassignment accuracy, which was not possible in the past using microsatellite markers. A baseline assessment of 395 walleye from 11 spawning stocks identified three reporting groups and refined previous assessments of gene flow among walleye stocks. Genetic assignment of 1,075 walleye harvested in eastern Lake Erie's recreational and commercial fisheries indicated that western basin stocks constituted the majority of harvest during the peak walleye fishing season (July-September), whereas eastern basin individuals comprised much of the early season harvest (May-June). Clear spatial structure in harvest composition existed; catches in more easterly sites contained more individuals of eastern basin origin than did more westerly sites. Our study provides important stock contribution estimates for Lake Erie fishery management and demonstrates the utility of genomic data to facilitate mixed-stock analysis in exploited fish populations having weak population structure or limited existing genetic resources.

Impacts of environmental matching on the routine metabolic rate and mass of native and mixed-ancestry brook trout (Salvelinus fontinalis) fry.

The environment an organism experiences during early development can impact its physiology and survival later in life. The objective of this study was to determine if temperatures experienced at embryonic life stages of brook trout (Salvelinus fontinalis) affected mass and routine metabolic rate (RMR) of a subsequent life stage (free-swimming fry). As part of this, we assessed the contributions and importance of hierarchical levels of biological organization [ancestral type (native vs. hatchery-introgressed), population, and family] to variability in mass and RMR of fry. As embryos and alevin, individuals were reared at either natural environmental (5°C) or elevated (9°C) temperatures and then acclimated to either matched or mismatched temperature treatments once yolk sacs were resorbed. Mass differences among fry were strongly influenced by population of origin as well as initial rearing and final acclimation temperatures. Variation in mass-adjusted RMR of fry was also strongly accounted for by source population, acclimation temperature, and individual mass. A significant interaction between population RMR and final acclimation temperature indicated that not all brook trout populations responded the same way to temperature changes. In contrast to expectations, the highest ancestry category (native vs. introgressed) did not significantly influence mass or mass-adjusted RMR.

Supplementation stocking of Lake Trout (Salvelinus namaycush) in small boreal lakes: Ecotypes influence on growth and condition.

Supplementation stocking is a commonly used management tool to sustain exploited fish populations. Possible negative consequences of supplementation on local stocks are a concern for the conservation of wild fish populations. However, the direct impacts of supplementation on life history traits of local populations have rarely been investigated. In addition, intraspecific hybridization between contrasting ecotypes (planktivorous and piscivorous) has been seldom considered in supplementation plans. Here, we combined genetic (genotype-by-sequencing analysis) and life history traits to document the effects of supplementation on maximum length, growth rates, body condition and genetic admixture in stocked populations of two Lake Trout ecotypes from small boreal lakes in Quebec and Ontario, Canada. In both ecotypes, the length of stocked individuals was greater than local individuals and, in planktivorous-stocked populations, most stocked fish exhibited a planktivorous-like growth while 20% of fish exhibited piscivorous-like growth. The body condition index was positively related to the proportion of local genetic background, but this pattern was only observed in stocked planktivorous populations. We conclude that interactions and hybridization between contrasting ecotypes is a risk that could result in deleterious impacts and possible outbreeding depression. We discuss the implications of these findings for supplementation stocking.

Metabolic rates of embryos and alevin from a cold-adapted salmonid differ with temperature, population and family of origin: implications for coping with climate change.

Early developmental stages of cold-adapted ectotherms such as brook trout (Salvelinus fontinalis) are at higher risk of mortality with increasing water temperatures. To determine the amount of variation present in early life, which may allow for potential adaptation to increasing temperature, we examined the routine metabolic rates (RMR) of wild-origin brook trout embryos and alevins reared at normal (5°C) and elevated (9°C) temperatures. The experiment was structured to examine variation in RMR within and among several levels of biological organization (family, population and ancestral type (native vs. mixed ancestry)). As expected, family and temperature variables were most important for predicting RMR and body mass, although population-level differences also existed when family was excluded for more detailed analysis. Additionally, body mass strongly influenced RMR at all life stages except for eyed embryos. When family identity was removed from the analysis, population became the most significant variable. Variation in RMR and mass within and among populations may indicate existing adaptive potential within and among brook trout populations to respond to predicted warming under climate change scenarios.

Acclimation capacity of the cardiac HSP70 and HSP90 response to thermal stress in lake trout (Salvelinus namaycush), a stenothermal ice-age relict.

The ability of cold-adapted species to persist in the face of climate change will depend on the capacity of individuals for thermal acclimation, adaptation, and the degree of physiological variation that exists among populations. We tested the acclimation capacity of cardiac HSP70 and HSP90 in four populations of lake trout (Salvelinus namaycush), a cold-adapted salmonid. We acclimated fish to four temperatures (8°C, 11°C, 15°C, and 19°C) and measured cardiac HSP70 and HSP90 levels by western blot prior to and following an acute (1h) heat shock at 23°C. Basal and induced cardiac HSP70 and HSP90 expression was similar among lake trout populations. Induced cardiac HSP70 was significantly lower in heat-shocked lake trout acclimated to 19°C compared to all other temperatures. Acclimation temperature had no significant effect on cardiac HSP90 prior to heat shock (control HSP90) and a significant effect on HSP90 in heat-shocked fish, although most significant treatment comparisons were marginal. However, for warm-acclimated fish (15°C and 19°C), cardiac HSP90 levels were lower in heat-shocked versus control fish. Together, this suggests that the induction temperature of cardiac HSP synthesis may increase with warm acclimation in lake trout. This plasticity in the cardiac HSR could assist lake trout populations in coping with longer periods of thermal stress as predicted by climate change models. The cardiac HSR supports previous research suggesting lake trout thermal physiology may be conserved across a wide geographic range.

Genetic mating system and mate selection in smallmouth bass.

Mating systems are an important factor influencing the variance in reproductive success among individuals within natural populations and thus have important ecological and evolutionary implications. We used molecular pedigree reconstruction techniques with microsatellite DNA data to characterize the genetic mating system and mate selection in adult smallmouth bass spawning in Lake Opeongo. The genetic mating system of smallmouth bass in this system can be characterized as predominantly monogamous with a low rate of polygynandry particularly among larger individuals. Iteroparous individuals showed a complete absence of interannual mate fidelity, presumably due to the low annual return rate of spawning adults. Within a season, individuals from both sexes pursued additional mating opportunities with males showing greater variance in mate number than females. Female mate selection appeared to be largely random with little evidence for elevated levels of inbreeding in this population. Multiple mating females pursued additional males to whom they were less related than the first male with which they spawned within a given season, however, this pattern varied among years. The mating pattern observed in this population would likely limit the strength of sexual selection and thus could account for the lack of sexual dimorphism and the absence of alternative reproductive tactics in this species.

Development and Validation of Environmental DNA (eDNA) Markers for Detection of Freshwater Turtles.

Environmental DNA (eDNA) is a potentially powerful tool for detection and monitoring of rare species, including threatened native species and recently arrived invasive species. Here, we develop DNA primers for a suite of nine sympatric freshwater turtles, and use it to test whether turtle eDNA can be successfully detected in samples from aquaria and an outdoor pond. We also conduct a cost comparison between eDNA detection and detection through traditional survey methods, using data from field surveys at two sites in our target area. We find that eDNA from turtles can be detected using both conventional polymerase chain reaction (PCR) and quantitative PCR (qPCR), and that the cost of detection through traditional survey methods is 2-10X higher than eDNA detection for the species in our study range. We summarize necessary future steps for application of eDNA surveys to turtle monitoring and conservation and propose specific cases in which the application of eDNA could further the conservation of threatened turtle species.

Intraspecific variation in thermal tolerance and acclimation capacity in brook trout (Salvelinus fontinalis): physiological implications for climate change.

Cold-water fishes are becoming increasingly vulnerable as changing thermal conditions threaten their future sustainability. Thermal stress and habitat loss from increasing water temperatures are expected to impact population viability, particularly for inland populations with limited adaptive resources. Although the long-term persistence of cold-adapted species will depend on their ability to cope with and adapt to changing thermal conditions, very little is known about the scope and variation of thermal tolerance within and among conspecific populations and evolutionary lineages. We studied the upper thermal tolerance and capacity for acclimation in three captive populations of brook trout (Salvelinus fontinalis) from different ancestral thermal environments. Populations differed in their upper thermal tolerance and capacity for acclimation, consistent with their ancestry: the northernmost strain (Lake Nipigon) had the lowest thermal tolerance, while the strain with the most southern ancestry (Hill's Lake) had the highest thermal tolerance. Standard metabolic rate increased following acclimation to warm temperatures, but the response to acclimation varied among strains, suggesting that climatic warming may have differential effects across populations. Swimming performance varied among strains and among acclimation temperatures, but strains responded in a similar way to temperature acclimation. To explore potential physiological mechanisms underlying intraspecific differences in thermal tolerance, we quantified inducible and constitutive heat shock proteins (HSP70 and HSC70, respectively). HSPs were associated with variation in thermal tolerance among strains and acclimation temperatures; HSP70 in cardiac and white muscle tissues exhibited similar patterns, whereas expression in hepatic tissue varied among acclimation temperatures but not strains. Taken together, these results suggest that populations of brook trout will vary in their ability to cope with a changing climate.

Does human-induced hybridization have long-term genetic effects? Empirical testing with domesticated, wild and hybridized fish populations.

Current conservation practices exclude human-generated hybridized populations from protection, as the genetic effects of hybridization in the wild have been observed to be long-lasting based on neutral genetic markers and are considered potentially irreversible. Theory, however, predicts otherwise for genes under selection. We transplanted combinations of wild, domesticated and hybridized populations of a fish species to new environments. We then compared survival, phenotypic variation and plasticity to determine whether hybridization affects adaptive potential after multiple generations of selection in the wild. Although the fitness of our hybridized populations at the onset of hybridization cannot be assessed, our results suggest that within five to eleven generations, selection can remove introduced foreign genes from wild populations that have hybridized with domesticated conspecifics. The end result is hybridized populations that, in terms of survival, phenotypic plasticity, mean trait expression and overall general responses to environmental change, closely resemble neighbouring wild populations. These results have important implications for considering the potential conservation value of hybridized populations and illustrate the effectiveness of selection in a local environment.

Ice age fish in a warming world: minimal variation in thermal acclimation capacity among lake trout (Salvelinus namaycush) populations.

In the face of climate change, the persistence of cold-adapted species will depend on their adaptive capacity for physiological traits within and among populations. The lake trout (Salvelinus namaycush) is a cold-adapted salmonid and a relict from the last ice age that is well suited as a model species for studying the predicted effects of climate change on coldwater fishes. We investigated the thermal acclimation capacity of upper temperature resistance and metabolism of lake trout from four populations across four acclimation temperatures. Individuals were reared from egg fertilization onward in a common environment and, at 2 years of age, were acclimated to 8, 11, 15 or 19°C. Although one population had a slightly higher maximal metabolic rate (MMR), higher metabolic scope for activity and faster metabolic recovery across all temperatures, there was no interpopulation variation for critical thermal maximum (CTM) or routine metabolic rate (RMR) or for the thermal acclimation capacity of CTM, RMR, MMR or metabolic scope. Across the four acclimation temperatures, there was a 3°C maximal increase in CTM and 3-fold increase in RMR for all populations. Above 15°C, a decline in MMR and increase in RMR resulted in sharply reduced metabolic scope for all populations acclimated at 19°C. Together, these data suggest there is limited variation among lake trout populations in thermal physiology or capacity for thermal acclimatization, and that climate change may impact lake trout populations in a similar manner across a wide geographical range. Understanding the effect of elevated temperatures on the thermal physiology of this economically and ecologically important cold-adapted species will help inform management and conservation strategies for the long-term sustainability of lake trout populations.

Genetic and ecological assessment of population rehabilitation: walleye in Lake Superior.

Sustainable management of exploited species is an ongoing challenge, particularly where populations have collapsed or been depleted by overharvest and habitat alteration. The walleye (Sander vitreus) population in Lake Superior's Black Bay historically supported more than 90% of the commercial walleye harvest from the entire lake, but collapsed in 1968 and has still not recovered despite long-term closure of the fishery. In an effort to rehabilitate this population, hatchery-origin walleye from exogenous sources were released into Black Bay between 2003 and 2005. We used individual-based analysis of genetic data collected between 2007 and 2010 to examine the contributions of different wild sources and hatchery stocking events to the contemporary walleye population in Black Bay. We found that 75% of the walleye in Black Bay originated from above- and below-barrier native populations in the Black Sturgeon River. The hatchery stocking events differed considerably in their effectiveness: the 2003 release of fry had no measurable contribution, whereas the 2004 and 2005 releases of fingerlings contributed 71% and 45% of the fish in their respective age classes. Hatchery and wild fish were similar in size, but hatchery fish rarely utilized the river habitat where Black Bay walleye historically spawned, and there was little genetic evidence of interbreeding or natural recruitment of stocked fish. Overall, our results suggest that restoring habitat connectivity to facilitate wild recruitment has greater potential than further exogenous stocking to contribute to the recovery of walleye in this system.

Ovarian fluid influences sperm performance in lake trout, Salvelinus namaycush.

The objectives of this study were to determine whether (i) the presence and concentration of ovarian fluid (OF) affects sperm performance traits, and (ii) variation in sperm performance traits is due to male identity, female identity, and/or male×female interactions in lake trout, Salvelinus namaycush. Spermatozoa from four males were activated in river water and OF from four females at two concentrations (10 and 15%). Presence of ovarian fluid influenced sperm traits; no differences were detected between 10 and 15% OF. Sperm traits varied depending on parental identity, such that sperm of some males perform better in the OF of all females and that in OF of some females sperm traits are higher for all males.