Environmental DNA (eDNA) is an increasingly valuable, non-invasive tool for monitoring freshwater fish biodiversity. While most eDNA studies rely on surface water sampling, few have compared the effectiveness of surface water, deep water, and sediment samples in lake ecosystems. This study evaluates these sampling approaches across five lakes in Alberta's Oil Sands Region, an area of economic and cultural significance where Indigenous communities are leading monitoring through Indigenous Community-Based Monitoring programs. Using species-targeted quantitative PCR (qPCR) assays, we assessed fish detection effectiveness to inform long-term monitoring strategies. Deep-water samples consistently yielded the highest species detection rates and eDNA copy numbers across all lakes, outperforming both surface water and sediment. Surface water showed moderate detection success, while sediment exhibited variable performance, occasionally detecting benthic or rare taxa absent in water samples. These findings demonstrate the value of eDNA for documenting baseline fish communities and identifying rare or at-risk species. Yet, effective integration into ongoing monitoring programs requires tailoring survey designs to the ecological traits and behaviors of target species, including habitat preferences, life history strategies, and movement patterns. Based on this exploratory work, we recommend a multi-habitat, multi-season sampling framework aligned with fish ecology and lake dynamics to maximize detection accuracy and achieve a more comprehensive representation of aquatic biodiversity.

ABSTRACT Sedimentary DNA (sedDNA), a form of environmental DNA (eDNA) shed by aquatic organisms and preserved in sediment, is crucial for reconstructing historical community compositions in aquatic ecosystems. In Cowpar Lake (Dene name: Doghostú), Alberta, a significant landslide event in the early 1940s CE impacted the lake's geochemistry and fish populations, as documented by Indigenous Knowledge from the Chipewyan Prairie First Nation and corroborated by targeted fish sedDNA analyses. The present study used 18S rRNA and cytochrome oxidase I (COI) genes for DNA metabarcoding of a sediment core from Cowpar Lake to assess the effect of the documented landslide and to reconstruct the historical community composition of eukaryotic functional trophic groups, including photoautotrophs, mixotrophs, parasites, and consumers. Between 1948 and 1956 CE, a notable shift in community composition occurred, with a decline in the alpha diversity of eukaryotic amplicon sequence variants. The increased primary productivity and terrestrial organic input post‐1950 is correlated with an increased diversity of phototrophs and mixotrophs, suggesting potential algal blooms. While parasite diversity remained stable, consumer diversity declined, likely due to increased microbial respiration of organic matter, reducing oxygen levels and making the lake less hospitable for consumers like whitefish, which eventually disappeared in the lake. The reconstructed eukaryotic community profiles from sedDNA were consistent with Indigenous Knowledge of natural changes around the lake. The present study highlights the potential of braiding sedDNA data with Indigenous Knowledge to reconstruct long‐term changes in aquatic communities, offering high‐resolution baseline data for environmental monitoring and a deeper understanding of how freshwater systems respond to natural and human‐induced impacts.

In the Alberta Oil Sands Region (AOSR), environmental stressors linked to oil sands industrial activity may have significant and species-specific impacts on local wildlife. This study evaluated the kynurenine–tryptophan ratio (KTR) as a potential biomarker for environmental exposure in longnose suckers (Catostomus catostomus) and white suckers (Catostomus commersonii) collected from various locations within the AOSR. The relationship between KTR and CYP1 enzyme activity (ethoxyresorufin-O-deethylase; EROD) was assessed alongside biometric indices, including gonadosomatic index (GSI), hepatic somatic index (HSI), and fat content. Both species exhibited increased EROD activity when exposed to oil sands natural deposits and potential industrial activity, indicating significant polycyclic aromatic compound (PAC) exposure. However, KTR changes were species-dependent: longnose suckers showed an inversely proportional relationship between KTR and EROD, while white suckers displayed a directly proportional correlation. Longnose suckers downstream of both municipal waste and industrial activity exhibited significant increases in GSI and fat content, with KTR varying more consistently by location rather than sex, suggesting that KTR may be a more reliable marker for location-based exposure. Species-specific differences in KTR and EROD relationships may be influenced by the distinct environmental requirements of each species, and their differing sensitivities to environmental conditions, including temperature, turbidity and flow conditions, during sampling periods. These findings illustrate the complexity of interpreting environmental biomarkers in wildlife and emphasize the need to consider ecological requirements and environmental conditions. Further research is necessary to validate this biomarker across different years and conditions and enhance its application in environmental monitoring and conservation efforts.

The recovery of DNA preserved in the sediment of aquatic systems (sedDNA) has provided short-term and long-term data on biological groups (e.g., bacteria, phytoplankton, zooplankton, and fishes) and has advanced our understanding of how environmental changes have affected aquatic communities. Here, we demonstrate the use of fish sedDNA from lake sediment cores in reconstructing the history of fish occurrences in four lakes within the Oil Sands region in Alberta, Canada. A general fish and eight targeted qPCR-based eDNA assays for freshwater fish species (cisco, lake whitefish, northern pike, chain pickerel, burbot, rainbow trout, lake trout, and walleye) were rigorously designed and validated. Utilizing these assays, we detected sedDNA from several fish species in sediment cores spanning over a century, which aligned with conventional fish surveys and historical records of human-mediated introductions. The use of fish sedDNA provided greater temporal resolution into the historical fish faunal records, bridging the knowledge gap that spanned from 100- to 150-year-old data. The present study also allowed documentation of human-mediated introductions of fish populations to the examined lakes. These findings can be used to refine native freshwater fish ranges and clarify the influence of human-mediated introductions on fish diversity in lakes, providing essential baseline data for conservation management and environmental impact assessments.

Due to its widespread use for the treatment of Type-2 diabetes, metformin is routinely detected in surface waters globally. Laboratory studies have shown that environmentally relevant concentrations of metformin can adversely affect the health of adult fish, with effects observed more frequently in males. However, the potential risk to wild fish populations has yet to be fully elucidated and remains a topic of debate. To explore whether environmentally relevant metformin exposure poses a risk to wild fish populations, the present study exposed wild fathead minnows (Pimephales promelas) to 5 or 50 mu g/L metformin via 2 m diameter in-lake mesocosms deployed in a natural boreal lake in Northern Ontario at the International Institute for Sustainable Development Experimental Lakes Area (IISD-ELA). Environmental monitoring was performed at regular intervals for 8-weeks, with fish length, weight (body, liver and gonad), condition factor, gonadosomatic index, liver-somatic index, body composition (water and biomolecules) and hematocrit levels evaluated at test termination. Metabolic endpoints were also evaluated using liver, brain and muscle tissue, and gonads were evaluated histologically. Results indicate that current environmental exposure scenarios may be sufficient to adversely impact the health of wild fish populations. Adult male fish exposed to metformin had significantly reduced whole body weight and condition factor and several male fish from the high -dose metformin had oocytes in their testes. Metformin- exposed fish had altered moisture and lipid (decrease) content in their tissues. Further, brain (increase) and liver (decrease) glycogen were altered in fish exposed to high -dose metformin. To our knowledge, this study constitutes the first effort to understand metformin's effects on a wild small -bodied fish population under environmentally relevant field exposure conditions.

Industrial development in Canada’s oil sands region influences the ambient environment. Some of these influences, such as the atmospheric deposition of emitted particles and gases are well-established using chemical indicators, but the effects of this process on bioindicators examined in field studies are less well-supported. This study used an extensive dataset available from 1997 to 2019, spatio-temporal modeling (Integrated Nested Laplace Approximation), and data on industrial and non-industrial covariates, including deposition patterns estimated using HYSPLIT (Hybrid Single Particle Lagrangian Integrated Trajectory) to determine if changes in sentinel fishes collected in streams from Canada’s Oil Sands Region were associated with oil sands industrial activity. While accounting for background variables (e.g., precipitation), estimated deposition of particles emitted from mine fleets (e.g., Aurora North), in situ stacks (e.g., Primrose and Cold Lake), mine stacks (e.g., Kearl), mine dust (e.g., Horizon), road dust (e.g., Muskeg River mine), land disturbance in hydrologically-connected areas, and wildfires were all associated with at least one fish endpoint. While many individual industrial stressors were identified, a specific example in this analysis parallels other work: the potential influence of emissions from both Suncor’s powerhouse and dust emitted from Suncor’s petroleum coke pile may both negatively affect fish health. Comparisons of fitted values from models with the estimated industrial effects and with deposition rates set to zero suggested some negative (and persistent) influences of atmospheric deposition at some locations, such as the gonadosomatic index (GSI) in the lower Muskeg and Steepbank rivers. While there is evidence of some large differences at individual locations the mean GSI and body condition estimates have improved throughout the region since the beginning of these collections in the late 1990s potentially highlighting improved environmental performance at the facilities, widespread enrichment effects, or interactions of stressors. However, mean liver-somatic indices have also slightly increased but remain low. These results, coupled with others suggest the utility of spatio-temporal approaches to detect the influence and effects of oil sands development at both local and regional scales.

The oil sands area of northern Alberta has river sediments that contain natural bitumen. Eggs and fish in these rivers may be exposed to bitumen-related chemicals early in life. This paper assesses a short embryo-larval fish exposure to oil sands sediment and follows the fish behaviour as they mature in clean water and examines their breeding success as adults (5 months afterwards). The three different oil sands river sediments tested were: a sediment collected outside of the bitumen deposit (tested at 3 g/L, Reference sediment from upstream Steepbank River site), and two sediments collected within the deposit (each tested at low (1 g/L) and high (3 g/L) concentrations). The sediments within the bitumen deposit were from the Ells and Steepbank (Stp) Rivers, and both contained significant total PAHs (>170 ng/g wet weight sediment) and alkylated PAHs (>4480 ng/g). Fish were exposed to these sediments for 21 days (as eggs and larval fish), and then transferred permanently to clean water to mature and breed. There was a significant decrease in the number of egg clutches produced by fish exposed early in life to Stp downstream high sediment (compared to Reference sediment). There was also a decrease in overall cumulative egg production, with fish from Stp downstream high sediment producing just over 1000 eggs in total while fish exposed to Ref sediment produced nearly 6900 eggs. The fish with reduced egg production were also less social than expected as they matured, and they had a lower % of early vitellogenic eggs in their ovaries. Overall, the exposure shows that a single, brief exposure during early life stages to natural bitumen can affect fish in adulthood. Naturally occurring bitumen-derived PAHs can reduce fish reproductive output by complex mechanisms, measurable as lower ovary maturity and changes in social behaviour.

Lake sediment serves as a natural archive of historical biological information. The use of sedimentary DNA (sedDNA), a form of environmental DNA (eDNA) shed by aquatic organisms and preserved in sediment, has been instrumental in reconstructing past faunal composition in aquatic communities. However, the low abundance of fish sedDNA and the often humic-rich nature of lake sediments create methodological challenges for the accurate detection of target sedDNA using quantitative polymerase chain reaction (qPCR)-based approaches. Herein, we present a consolidated qPCR-based eDNA workflow to reconstruct past and current fish fauna in Cowpar Lake located in the Oil Sands region in Alberta (Canada), which were then validated using Indigenous Knowledge from Chipewyan Prairie First Nation community members. The present study highlights the importance of combining column- and precipitation-based PCR inhibitor clean-up, nucleic acid concentration, incorporating endogenous chloroplast DNA as a sample integrity control. Robust qPCR-based eDNA assays were also useful in preventing the false-negative detection of low copies of target fish DNA. The presence of Northern pike (1905 to 2019) and Cisco (1919 to 1942) in Cowpar Lake was confirmed based on detected sedDNA from sediment core. The reconstructed fish records from sedDNA-inferred data aligned with the Indigenous accounts of natural and human-mediated changes in land use around the lake. Overall, the present study addresses common methodological concerns in processing lake sediment samples for fish eDNA detection and demonstrates the great potential of combined eDNA-inferred data and Indigenous Knowledge in reconstructing historical fish records in aquatic communities.

Municipal wastewater treatment plant (WWTP) effluent is one of several point sources of contaminants (nutrients, pharmaceuticals, estrogens, etc.) which can lead to adverse responses in aquatic life. Studies of WWTP effluent impacts on rainbow darter (Etheostoma caeruleum) collected downstream of WWTPs in the Grand River, Ontario have reported disruption at multiple levels of biological organization, including altered vitellogenin gene expression, lower levels of in vitro steroid production, and high frequency of intersex. However, major upgrades have occurred at treatment plants in the central Grand River over the last decade. Treatment upgrades to the Waterloo WWTP were initiated in 2009 but due to construction delays, the upgrades came fully on-line in 2017/2018. Responses in rainbow darter have been followed at sites associated with the outfall consistently over this entire time period. The treatment plant upgrade resulted in nitrification of effluent, and once complete there was a major reduction in effluent ammonia, selected pharmaceuticals, and estrogenicity. This study compared several key responses in rainbow darter associated with the Waterloo WWTP outfall prior to and post upgrades. Stable isotopes signatures in fish were used to track exposure to effluent and changed dramatically over time, corresponding to the effluent quality. Disruptions in in vitro steroid production and intersex in the darters that had been identified prior to the upgrades were no longer statistically different from the upstream reference sites after the upgrades. Although annual variations in water temperature and flow can potentially mask or exacerbate the effects of the WWTP effluent, major capital investments in wastewater treatment targeted at improving effluent quality have corresponded with the reduction of adverse responses in fish in the receiving environment.

For over a decade, intersex has been observed in rainbow darter (RD) (Etheostoma caeruleum) populations living downstream wastewater treatment plants (WWTPs) in the Grand River, Ontario, Canada. To further our understanding of intersex development in adult male fish, the current study addressed three objectives: i) can intersex be induced in adult male fish, ii) is there a specific window of exposure when adult male fish are more susceptible to developing intersex, and iii) can pre-exposed adult male fish recover from intersex? To assess intersex induction in adult male fish, wild male RD were exposed in the laboratory for 22 weeks (during periods of spawning, gonadal regression, and gonadal recrudescence) to environmentally relevant concentrations of 17 & alpha;ethinylestradiol (EE2) including nominal 0, 1, and 10 ng/L. Intersex rates and severity at 10 ng/L EE2 were similar to those observed historically in adult male populations living downstream WWTPs in the Grand River and confirmed previous predictions that 1-10 ng/L EE2 would cause these adverse effects. To assess a window of sensitivity in developing intersex, male RD were exposed to nominal 0, 1 or 10 ng/L EE2 for 4 weeks during three different periods of gonadal development, including (i) spawning, (ii) early recrudescence and (iii) late recrudescence. These short-term exposures revealed that intersex incidence and severity were greater when RD were exposed while gonads were fully developed (during spawning) compared to periods of recrudescence. To assess if RD recover from intersex, wild fish were collected downstream WWTPs in the Grand River and assessed for intersex both before and after a 22-week recovery period in clean water that included gonadal regression and recrudescence. Results showed that fish did not recover from intersex, with intersex rates and severity similar to those both before and after the transition to clean water. This study further advances our knowledge on intersex manifestation in adult male fish including their sensitivity to endocrine active compounds during different periods of their annual reproductive cycle and their limited ability to recover from intersex after onset of the condition.

Fish consumption advisories for mercury (Hg) are common in rivers, highlighting connections between landscape sources of Hg and downstream fluvial ecosystems. Though watershed conditions can influence concentrations of Hg in smaller streams, how Hg changes downstream through larger rivers and how these changes associate with Hg concentrations in fish is not well understood. Here we present a continuum of concentrations and yields of total mercury (THg) and methylmercury (MeHg) from small tributary systems draining diverse western Canadian headwater landscapes through to major transboundary rivers. We associate these downstream patterns with THg concentrations in tissues of resident fish in major rivers. Mean concentrations and yields of unfiltered THg from over 80 monitored tributaries and major rivers were highly variable in space ranging from 0.28 to 120 ng L −1 and 0.39 to 170 µg ha −1 d −1 , respectively. Using spatial data and a hierarchical cluster analysis, we identified three broad categories of tributary catchment conditions. Linear mixed modeling analysis with water quality variables revealed significantly lower THg concentrations in tributaries draining cordillera-foothills (geometric mean: 0.76 ng L −1 ) regions relative to those draining forested (1.5 ng L −1 ) and agriculturalized landscapes (2.4 ng L −1 ), suggesting that sources and mobility of THg in soils and surface waters were different between landscapes. However, these concentration differences were not sustained downstream in major rivers as local sources and sinks of THg in river channels smoothed differences between landscape types. Extensive fish tissue monitoring in major rivers and ANCOVA analysis found that site-specific, river water THg and MeHg concentrations and local catchment conditions were stronger associates of THg concentrations in fish than broader trends in rivers within and across landscape classes. Consequently, site-specific, targeted monitoring of THg and MeHg concentrations in water and fish is a preferred study design when assessing regional-level patterns in fish tissue concentrations.

The collection of sentinel fish species for Environmental Effects Monitoring (EEM) has provided ancillary fish assemblage surveys on several tributaries in the Athabasca Oil Sands Region (AOSR) of Alberta, Canada over the last decade. Using available, comparable data we investigated baseline fish assemblage variability along the Ells River, a tributary of the Athabasca River experiencing increasing proximity to natural bitumen deposits and proposed mining development as it approaches confluence with the mainstem Athabasca. Transect-based electrofishing data from four sites surveyed in September 2013, 2014 and 2018 showed significant spatiotemporal variability in assemblages, where spatial variability was greatest in 2013 and temporal variability was observed in assemblages both upstream (2013–2014) and downstream (2014–2018) of proposed development. Habitat assessments in 2018 revealed significant relationships among pH, algae cover and site slope with fish assemblages of the same year. Due to the complementary nature of assemblage surveys, data challenges (changing methodologies, sampling effort, and limited ancillary physiochemical data) have presented limitations to the multivariate approach applied in the study. Moving forward, employing consistent methods for fish collections and fine-scale habitat assessments will improve the ability to correlate assemblage variability with changes in the physical environment. Ultimately, this will aid in developing potential triggers of change that may be attributed to or confound adjacent, expanding Oil Sands activities. These findings will also inform monitoring programs on the use of fish assemblages as indicators of change, potentially providing an alternative to existing biomonitoring approaches in small streams with small fish populations.

Metformin is routinely detected in aquatic ecosystems because of its widespread use as a treatment for Type 2 diabetes. Laboratory studies have shown that exposure to environmentally relevant concentrations of metformin can alter metabolic pathways and impact the growth of early life stage (ELS) fish; however, it is unknown whether these effects occur in wild populations. Herein, we evaluate whether findings from laboratory studies are representative and describe the relative sensitivities of both populations. Duplicate exposures (0, 5, or 50 μg/L metformin) were conducted using wild- and lab-spawned fathead minnow (Pimephales promelas) embryos. Apart from the water source, exposure conditions remained constant. Wild embryos were exposed to previously dosed lake water to account for changes in bioavailability, while reconstituted freshwater was used for the laboratory study. Developmental metformin exposure differentially impacted the growth and morphology of both cohorts, with energy dyshomeostasis and visual effects indicated. The fitness of wild-spawned larvae was impacted to a greater extent relative to lab-spawned fish. Moreover, baseline data reveal important morphological differences between wild- and lab-spawned ELS fatheads that may diminish representativeness of lab studies. Findings also confirm the bioavailability of metformin in naturally occurring systems and suggest current exposure scenarios may be sufficient to negatively impact developing fish.

Industrial and other human activities in Canada’s oil sands region (OSR) influence the environment. However, these impacts can be challenging to separate from natural stresses in flowing waters by comparing upstream reference sites to downstream exposure locations. For example, health indicators of lake chub (Couesius plumbeus) compared between locations in the Ells River (Upper and Lower) in 2013 to 2015 and 2018 demonstrated statistical differences. To further examine the potential sources of variation in fish, we also analyzed data at sites over time. When fish captured in 2018 were compared to pooled reference years (2013–2015), results indicated multiple differences in fish, but most of the differences disappeared when environmental covariates were included in the Elastic Net (EN) regularized regression models. However, when industrial covariates were included separately in the EN, the large differences in 2018 also disappeared, also suggesting the potential influence of these covariables on the health of fish. Further ENs incorporating both environmental and industrial covariates along with other variables which may describe industrial and natural influences, such as spring or summer precipitation and summer wind speeds and distance-based penalty factors, also support some of the suspected and potential mechanisms of impact. Further exploratory analyses simulating changes from zero and the mean (industrial) activity levels using the regression equations respectively suggest effects exceeding established critical effect sizes (CES) for fish measurements may already be present or effects may occur with small future changes in some industrial activities. Additional simulations also suggest that changing regional hydrological and thermal regimes in the future may also cause changes in fish measurements exceeding the CESs. The results of this study suggest the wide applicability of the approach for monitoring the health of fish in the OSR and beyond. The results also suggest follow-up work required to further evaluate the veracity of the suggested relationships identified in this analysis.

Liver and skin tumours in brown bullhead (Ameiurus nebulosus) and white sucker (Catostomus commersoni) have been associated with contamination of the aquatic environment. In this study, we present a Bayesian hierarchical logistic-Bernoulli framework that considers the role of fish covariates (e.g., age, total weight, gonad weight, liver weight, fork length) on tumour prevalence to identify the presence of “hot-spots” around the Canadian waters of the Great Lakes, where high fish tumour frequencies are registered. We developed methods to discern the degree of impairment that are either based on the comparison of tumour frequencies in contaminated (or impacted) sites against those predicted in their corresponding reference areas, or the assessment of the prevailing conditions in impaired sites independently, without the need to establish baseline conditions for comparison purposes. Our modelling study predicts low frequencies of neoplastic tumours in all the impacted locations. In contrast, the same comparisons with the preneoplastic lesions provided evidence of distinct differences between impacted and reference sites in Jackfish Bay, St. Mary’s River, Niagara River, Hamilton Harbour, and Bay of Quinte. We also found weak to moderately strong relationships between tumour occurrence and fish physical characteristics that varied considerably in terms of their strength and nature (sign) among the different locations. Our study concludes that the prevalence of neoplastic tumours appears to have reached acceptable levels around the Great Lakes, but the distinctly higher levels of preneoplasms in several impacted locations underscore the need to improve our understanding of the lesions that may lead to carcinogenesis.

Effluents from municipal wastewater treatment plants (MWTPs) are complex mixtures of chemicals including endocrine-disrupting compounds (EDCs) and 17α-ethynylestradiol (EE2). The objective of this study was to evaluate selected responses of two fish species, in two different years, exposed in situ to MWTP effluent. Biological markers of exposure (plasma vitellogenin (VTG) and antioxidant enzymes) were measured in two species of male fish, rainbow trout (Oncorhynchus mykiss) and fathead minnow (Pimephales promelas), caged at sites associated with wastewater outfall. The estrogenicity of the final effluent in 2010 was determined to be 17.0 + 0.4 ng/L estrogen equivalents (EEQ) and reduced to 7.5 + 2.9 ng/L EEQ after infrastructure upgrades. Pharmaceuticals and personal care products in the effluent and surface water in both years confirmed the exposures at each downstream site. Despite the presence of estrogenic compounds in the MWTP effluent, no effluent-caged male fish demonstrated plasma VTG induction. Minnows and trout that received an intraperitoneal injection of 5 mg/g EE2 showed VTG induction at both field sites. In 2012, the liver somatic index (LSI) of both species increased with exposure, as did changes in antioxidant enzymes, and reactive oxygen species (ROS) activity. Multiple biological mechanisms are modified by effluent exposure, and multiple endpoints are needed to assess risk.

Jackfish Bay is an isolated bay on the north shore of Lake Superior, Canada that has received effluent from a large bleached-kraft pulp mill since the 1940s. Studies conducted in the late 1980s found evidence of reductions in sex steroid hormone levels in multiple fish species living in the Bay, and increased growth, condition and relative liver weights, with a reduction in internal fat storage, reduced gonadal sizes, delayed sexual maturation, and altered levels of circulating sex steroid hormones in white sucker ( Catostomus commersonii ). These early studies provided some of the first pieces of evidence of endocrine disruption in wild animals. Studies on white sucker have continued at Jackfish Bay, monitoring fish health after the installation of secondary waste treatment (1989), changes in the pulp bleaching process (1990s), during facility maintenance shutdowns and during a series of facility closures associated with changing ownership (2000s), and were carried through to 2019 resulting in a 30-year study of fish health impacts, endocrine disruption, chemical exposure, and ecosystem recovery. The objective of the present study was to summarize and understand more than 75 physiological, endocrine, chemical and whole organism endpoints that have been studied providing important context for the complexity of endocrine responses, species differences, and challenges with extrapolation. Differences in body size, liver size, gonad size and condition persist, although changes in liver and gonad indices are much smaller than in the early years. Population modeling of the initial reproductive alterations predicted a 30% reduction in the population size, however with improvements over the last couple of decades those population impacts improved considerably. Reflection on these 30 years of detailed studies, on environmental conditions, physiological, and whole organism endpoints, gives insight into the complexity of endocrine responses to environmental change and mitigation.

The St. Clair River, which flows from Lake Huron to Lake St Clair, receives discharges and emissions from numerous petroleum refineries, petrochemical facilities, and other sources. We used a multi-factor ANOVA to test for spatial and temporal changes in concentrations of trace metals/metalloids in homogenates of fish from Lake Huron (reference site) and two river sites - an industrialized region (Stag Island) and downstream Walpole Island/Chenal Écarte. In 2002/2003 and in 2014, we sampled 20 adult fish of each sex of the following species: shorthead redhorse (Moxostoma macrolepidotum), yellow perch (Perca flavescens); we added emerald shiner (Notropis atherinoides; unsexed) in 2014. There was a temporal decline of most metals/metalloids in shorthead redhorse at the river sites. Linear Discriminant Analysis separated the 2014 shorthead redhorse by site. Emerald shiners had higher concentrations of metals/metalloids, apart from mercury, than the other species; those concentrations were highest at Chenal Écarte, which warrants further research. Tissue concentrations of mercury, which declined temporally at Stag Island and Walpole Island, were above the protective guidance for wildlife consumers at all sites, but were lower than predicted to affect fish health. Apart from increased liver size (ANCOVA) in shorthead redhorse at Walpole Island (2014), the health variables in the riverine fish were stable or differed slightly from reference, which indicated good health and reproductive potential. In shorthead redhorse, tissue metals correlated inconsistently with health indices. Further research could ascertain if temporal declines in metals/metalloids in riverine SHRH are benefits of remediation actions or fluctuations with other causes.