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The relationships between fecundity and size of Atlantic herring (Clupea harengus) were estimated within five different spawning areas off the coast of Nova Scotia in 2019 and 2020. Statistically significant differences in fecundity relative to body weight were observed among spawning areas and between years. Fecundity-at-length on the German Bank spawning ground was 29%–36% and 22%–28% lower than estimates from 2001 and 1970, respectively. Temporal changes in weight- and relative fecundity-at-age resulted in a decrease in the number of eggs-per-recruit (in an equilibrium unfished state) by 50% and a decrease of 27% in the egg production per tonne of spawning stock biomass (SSB) in 2020 relative to 1970. Decreases in SSB-per-recruit and eggs-per-recruit over time resulted in proportional decreases in equilibrium SSB at maximum sustainable yield (MSY); however, the fishing mortality rate (F) at MSY remained relatively stable over time. Total egg production was shown to be disproportional to SSB. Equilibrium SSB at MSY was greater (and F at MSY lower) when estimated using eggs-per-recruit compared to SSB-per-recruit. Failing to account for fecundity and assuming that egg production is proportional to SSB resulted in an overestimate of stock status.

Many present-day aquatic ecosystems are defined by anthropogenic activities such as the introduction of non-native species, habitat loss, and eutrophication. On Prince Edward Island, Canada, non-native rainbow trout (Oncorhynchus mykiss) have established in highly agricultural coastal watersheds. We used acoustic telemetry with continual oxygen and temperature monitoring to explore the relationship between rainbow trout movements, low dissolved oxygen, high temperature, and other environmental parameters in the eutrophic Dunk River estuary. We tested the hypothesis that rainbow trout capitalize on increased productivity from nutrient stimulation but face a trade-off with disturbed water quality. We divided the salinity transition gradient into zones representing the transition from freshwater to seawater and established that temperature and hypoxia increased with salinity. During hypoxic periods, movements decreased, and residence increased in low salinity habitat ( 6 mg L−1, discharge averaging 2 m3 s−1, and temperatures below 23 °C. This study provides insights into the success of rainbow trout in eutrophic estuaries.

Agricultural activity within coastal watersheds results in estuaries becoming the receiving environment for pesticide inputs. In estuaries, salinity can alter insecticide responses of exposed crustaceans. The acute toxicity of environmentally relevant doses of chlorpyrifos and imidacloprid were examined using the euryhaline amphipod Gammarus lawrencianus at 20 and 30 Practical Salinity Units (PSU). Responses were recorded every 24 h until an incipient (threshold) L(E)C50 was reached. For chlorpyrifos, LC50 ranged from 0.1 to 0.5 µg/L and was two-fold higher at 30 vs. 20 PSU at all time-points over the 96 h exposure. Imidacloprid immobility EC50 ranged from 4 to 40 µg/L over the 144 h exposure. An effect of salinity was only observed at 48 h and the EC50 values showed 1.4 times more potency at 20 PSU compared to 30 PSU. Measured concentrations of both compounds did not differ between salinities. Acetylcholinesterase activity in chlorpyrifos exposed amphipods showed no salinity effect at 96 h. We conclude that salinity level alters G. lawrencianus susceptibility to chlorpyrifos exposure, but not imidacloprid.

The common bully (Gobiomorphus cotidianus) was used as a bioindicator to assess the impacts of geothermal, bleached kraft mill effluent (BKME), sewage, and thermal discharges in the Waikato River, Northern New Zealand. A range of morphological and physiological parameters were measured to assess fitness in fish sampled upstream and downstream from these point sources. Isotopic signatures gave evidence that fish were resident at sites and showed a gradient corresponding to site-specific sources of carbon and nitrogen along the river. There were changes in the size range and age structure of the populations but, no physiological responses were detected. Limited impacts were observed on common bully health at individual discharge sites, or downstream where the river showed gradual deterioration in water quality, particularly nutrient enrichment, and increased turbidity. Despite the gradual deterioration in water quality no concomitant cumulative impacts were observed in common bully. Responses were largely local in nature, responding to point-source discharges. Population responses at the geothermal and BKME discharge sites warrant further study to investigate possible effects on fish recruitment.

This study provides evidence of two subpopulations of Atlantic halibut (Hippoglossus hippoglossus) in the southern Gulf of St. Lawrence. The migrations of 20 Atlantic halibut captured in the coastal waters of Prince Edward Island, Canada, were evaluated using pop-up satellite archival tags (PSAT). Geolocation data showed that Atlantic halibut migrated north to the Laurentian Channel via distinct eastern or western routes. Floy tagging and recapture (recapture of 18.4%) showed that less than 7.0% of Atlantic halibut exhibited dispersive behaviour outside of their annual migratory route. Overwintering occurred on the slope and in the deepest waters of the Laurentian Channel. Atlantic halibut in the deepest waters of the Laurentian Channel exhibited rapid, ∼100 m rises, presumed to be associated with spawning from January to March. The eastern and western migratory cohorts exhibited this behaviour ∼350 km apart, suggesting reproductive isolation as the basis of subpopulations. The results of this study indicate a need to reconsider the management of Gulf of St. Lawrence Atlantic halibut as one continuous population.

This study examined the migratory patterns of introduced rainbow trout (Oncorhynchus mykiss) in three rivers in Prince Edward Island, Canada, using acoustic telemetry and otolith microchemistry. Only 6% of acoustically tagged fish in three river systems left coastal embayments. A cohort of rainbow trout in all three rivers entered saline waters. Habitat use differed among migrants in the three rivers, as Montague River fish occupied estuary habitat (mean 20.79 PSU) more often than West River and Dunk River fish that tended to occupy both riverine tidal (mean 1.27 and 4.29 PSU, respectively) and freshwater habitats (

This study examined the responses of a population of brook stickleback (Culaea inconstans) exposed to pulp mill effluent at Jackfish Bay, Lake Superior, Canada, in May 2007 and May 2011. Brook stickleback were extirpated from the effluent-receiving site, presumably due to anoxia after this period. Females at the effluent-receiving site had significantly larger gonad sizes in 2007 and 2011. In 2011, effluent-exposed female gonadal development was significantly advanced when compared with reference sites; they were the second most mature when compared among three different reference sites. Analysis of 7-ethoxyresorufin-O-deethylase activity revealed that effluent-receiving site females had greater CYP1A induction in 2007 and significantly greater CYP1A induction in 2011. Effluent-receiving site males showed significantly reduced CYP1A induction in 2007 and significantly greater induction in 2011. Chemical evaluation of sediment from the receiving environment showed elevated levels of resin acids and the polycyclic aromatic hydrocarbon, retene. Higher condition factors and more mature gonads were consistent with higher winter and spring temperatures modified by effluent or by lake vs. stream environments. Overall, effects on effluent-exposed brook stickleback were not consistent with reported effects in white sucker exposed to the same effluent in previous studies. This article has been made Open Access thanks to the kind support of CAWQ/ACQE (https://www.cawq.ca).

Simulation of surface water flow and temperature under a non-stationary, anthropogenically impacted climate is critical for water resource decision makers, especially in the context of environmental flow determination. Two climate change scenarios were employed to predict streamflow and temperature: RCP 8.5, the most pessimistic with regards to climate change, and RCP 4.5, a more optimistic scenario where greenhouse gas emissions peak in 2040. Two periods, 2018–2050 and 2051–2100, were also evaluated. In Canada, a number of modelling studies have shown that many regions will likely be faced with higher winter flow and lower summer flows. The CEQUEAU hydrological and water temperature model was calibrated and validated for the Wilmot River, Canada, using historic data for flow and temperature. Total annual precipitation in the region was found to remain stable under RCP 4.5 and increase over time under RCP 8.5. Median stream flow was expected to increase over present levels in the low flow months of August and September. However, increased climate variability led to higher numbers of periodic extreme low flow events and little change to the frequency of extreme high flow events. The effective increase in water temperature was four-fold greater in winter with an approximate mean difference of 4 °C, while the change was only 1 °C in summer. Overall implications for native coldwater fishes and water abstraction are not severe, except for the potential for more variability, and hence periodic extreme low flow/high temperature events.

Many environmental monitoring programs include an assessment of the health of fish populations using a sentinel species and include an indicator of reproductive potential. Knowledge of the reproductive strategy of the fish species is critical for data interpretation but is not always known. The reproductive strategy of a species can be determined from detailed histological analyses of ovaries throughout the reproductive cycle; however, these studies can be costly and can delay the implementation of a monitoring program. Three quick and cost-effective methods of predicting the reproductive strategy (annual single spawning or annual multiple spawning) are evaluated in this study using predicted probabilities from binary logistic regression models as a means of classifying the reproductive strategies of 18 different fish species in Atlantic Canada. The first method was based on the hypothesis that the variability in the ovary weight-body weight relationship in prespawning females is higher in multiple spawners. This method did not have a good classification rate due to some multiple spawners having low variability. The other two methods involved predictor variables representing the proportion of oocytes in different stages of development and predictor variables representing the distribution of oocyte sizes during the prespawning season for 111 fish (25 different samples for species). Predicted probabilities from these regression models could be used to correctly classify the reproductive strategies of all 25 samples (development stage model) and all but one sample (oocyte size distribution model). These models can be used to estimate the reproductive strategy of a species from a single sample of fish collected during the prespawning period to support species selection and data interpretation in environmental monitoring programs.

Agricultural activity within coastal watersheds results in estuaries becoming the receiving environment for pesticide inputs. In estuaries, salinity can alter insecticide responses of exposed crustaceans. The acute toxicity of environmentally relevant doses of chlorpyrifos and imidacloprid were examined using the euryhaline amphipod Gammarus lawrencianus at 20 and 30 Practical Salinity Units (PSU). Responses were recorded every 24 h until an incipient (threshold) L(E)C

Movement of anadromous fishes from marine to freshwater habitats provides a seasonal pulse of marine-derived nutrients (MDNs) to freshwater biota. In northeastern North America, iteroparous...

The purpose of this study was to update rainbow trout (Oncorhynchus mykiss) invasion status, delineate factors that increase the invasion probability, and quantify habitat overlap between invasive rainbow trout and native Atlantic salmon (Salmo salar) and brook trout (Salvelinus fontinalis) on Prince Edward Island, Canada. Analysis of landscape-level variables in 26 watersheds (14 with and 12 without rainbow trout) demonstrated that watershed slope, percent agricultural land use, and distance to the nearest rainbow trout population significantly increased the probability of rainbow trout establishment. Two independent reach-level fish community surveys were conducted by electrofishing in combination with habitat surveys. Reaches with rainbow trout had higher stream slope, Atlantic salmon were found in wider reaches with larger substrate, and brook trout were typically dominant in headwater reaches with finer substrate. Findings at multiple ecological scales illustrated that rainbow trout invasion success is positively influenced by the presence of high-slope habitat. Habitat separation of the three salmonid species indicates that competition with introduced rainbow trout may not be the most significant threat to native salmonid populations.

Naphthenic acids (NAs) are predominant compounds in oil sands influenced waters. These acids cause numerous acute and chronic effects in fishes. However, the mechanism of toxicity underlying these effects has not been fully elucidated. Due to their carboxylic acid moiety and the reported disruption of cellular bioenergetics by similar structures, we hypothesized that NAs would uncouple mitochondrial respiration with the resultant production of reactive oxygen species (ROS). Naphthenic acids were extracted and purified from 17-year-old oil sands tailings waters yielding an extract of 99% carboxylic acids with 90% fitting the classical O2–NA definition. Mitochondria were isolated from rainbow trout liver and exposed to different concentrations of NAs. Mitochondrial respiration, membrane potential, and ROS emission were measured using the Oroboros fluorespirometry system. Additionally, mitochondrial ROS emission and membrane potential were evaluated with real-time flow cytometry. Results showed NAs uncoupled...

Simple empirical models can sometimes capture salient patterns without sacrificing predictive capacity when compared to more complex models. Herein we examine dissolved oxygen as an indicator of eutrophication status for shallow estuaries. Dissolved oxygen was measured hourly in the upper estuary of 15 watersheds along a nutrient-loading and geographic gradient. Metrics describing hypoxia and supersaturation were devised and then analyzed using multivariate statistics. Results revealed independent responses for hypoxia and supersaturation with hypoxia-related metrics correlating strongly with water residence. A metric integrating hypoxia and supersaturation effectively discriminated between seagrass and algae-dominated habitats and was significantly correlated with both water residence and nitrate-N loading. Chlorophyll, measured bi-weekly, was not correlated with our predictor variables likely because it does not account for benthic production. Over 70% of the variability in hypoxia was explained by water residence and nitrate-N loading indicating that this model can be of use for managers.

Chlorpyrifos is an organophosphate that is currently used to reduce arthropod pests for the protection of agricultural crops. Coastal marine ecosystems may be exposed to agricultural pesticides via runoff and pesticide exposure can impact the health and survival of non-target species such as the American lobster (Homarus americanus). In the current study, the gene expression changes of H. americanus stage IV larvae were evaluated to understand the physiological mechanisms affected by exposure to sublethal concentrations of chlorpyrifos. After 48 h chlorpyrifos exposure, surviving lobsters were processed for Illumina RNA sequencing (RNA-seq). Genes of interest that showed significant changes using RNA-seq were verified using reverse transcriptase quantitative polymerase chain reaction (RT-qPCR). Analysis of RNA-seq and the confirmation of gene expression patterns via RT-qPCR found altered expression in genes related to stress response (glutathione peroxidase 3 and heat shock protein 60), hypoxia response (hairy, astakine 2, hemocyanin), moulting (cytochrome P450 307a1 and chitinase), and immunity (astakine 2) pathways. Changes to gene expression were most notable in lobsters exposed to 0.57 μg/L chlorpyrifos.

A data-driven model based on an adaptive neuro-fuzzy inference system (ANFIS) was tested for the estimation of suspended sediment concentrations within watersheds influenced by agriculture. ANFIS models were developed using different combinations of inputs such as precipitation, streamflow, surface runoff and the watershed vulnerability index. A multi-watershed ANFIS model was also developed combining the datasets from all studied watersheds. The best results were obtained from a combination of precipitation, streamflow and watershed vulnerability index as input variables. Nash-Sutcliffe coefficients were improved for the multi-watershed ANFIS compared to watershed-specific ANFIS models. The introduction of the erosion vulnerability index significantly improved the ability of the ANFIS model to estimate suspended sediment concentrations within the watersheds. Furthermore, the inclusion of this index opens the possibility of using the ANFIS model to investigate the impact of land-use changes on sediment delivery.

Seagrasses are declining globally; understanding and monitoring these trends forms an important basis for management practices. This study examined eelgrass (Zostera marina) structural and isotopic indicators as a means of characterizing stress from the impacts of agricultural land use in four estuaries in the Southern Gulf of St. Lawrence, Canada. These estuaries ranged from 5 to 62% agricultural land use in their respective watersheds. Anthropogenic stressors such as increased nutrient loading and decreased light penetration (from sedimentation and phytoplankton) and natural stressors such as low salinity and temperature were measured concurrently with plant structural indicators, e.g., aboveground biomass, belowground biomass, canopy height, and root-to-shoot ratio, across the spatial range of eelgrass presence in each estuary. Plant isotopic indicators included leaf tissue concentrations of C, N, S, and stable isotopes thereof and were measured at each station in eelgrass leaf tissue. Eelgrass plant indicators varied significantly across the gradients within each estuary as well as throughout the growing season, but due to the very high within-estuary variability indicators did not vary significantly between estuaries. Eelgrass leaf nutrients and stable isotopes also varied significantly between stations within estuaries though in this case, among-estuary differences were detected. A distance-based linear model was employed to examine the ability of estuarine characteristics and plant isotopic and elemental variables to predict the plant structural indicators. Results showed that factors related to both salinity and nitrogen loading influenced the biomass and structure of this plant but in different ways. Belowground biomass most correlated to salinity, and aboveground biomass most related to nitrogen loading with a strong relationship with light attenuation. Overall, the high level of within-estuary variability, and the strong influence of salinity, suggests that comparison of plant structural indicators between estuaries may have limited utility as a monitoring tool; eelgrass coverage throughout an estuary may better reflect the influence of land use at a regional scale.

Biological and chemical endpoints were measured in white sucker collected downstream of Athabasca oil sands developments (AB, Canada) and compared with those at Calling Lake (AB, Canada), a reference location upstream of the Athabasca oil sands deposit. Naphthenic acid concentrations were also measured at 14 sites in the Athabasca River watershed. Concentrations of naphthenic acids were elevated in tributaries adjacent to oil sands mining developments. Tributary naphthenic acid profiles were more similar to aged oil sands process water than samples from the Athabasca River, suggesting an influence of tailings in the tributaries. White sucker showed higher energy storage in the Athabasca River as indicated by significantly higher condition and liver size. White sucker were not investing that energy into reproductive effort as measured by gonad size and fecundity, which were significantly reduced relative to the reference location. White sucker showed increased exposure to polycyclic aromatic hydrocarbons as indicated by hepatic cytochrome P4501A (CYP1A) activity and fluorescent bile metabolites, as well as higher concentrations of naphthenic acids in bile. Cadmium, copper, nickel, and selenium were also elevated in white sucker liver tissue compared with the reference location. Based on the exposure profile and response pattern observed, effects on energy storage and utilization in white sucker from the Athabasca River most likely resulted from exposure to polycyclic aromatic hydrocarbons derived from petrogenic and pyrolytic sources. Environ Toxicol Chem 2017;36:2058-2067. © 2017 SETAC.

The increased soil loss in an agricultural watershed raises challengers for river water quality and a reliable automated monitoring for suspended sediment concentrations (SSC) is crucial to evaluate sediment budgets variation in systems. The aims of this study were (1) to test if an acoustic doppler current profiler (ADCP) would give similar results to turbidity probe measurements as a high frequency monitoring tool for suspended sediment, and (2) to analyze the relationship between sediment drivers and SSC in a typical agricultural drainage basin. The acoustic and optical backscatter sensors were used to collect SSC data during the ice-free seasons of four consecutive years in the Dunk River (PEI, Canada). The slopes of the relationships between the two SSC indirect measurements were not significantly different than 1. Correlations between SSC and hydro-meteorological variables showed that the high SSC values were more associated with the streamflow and water velocity than precipitation. This study highlighted the great potential of ADCP for the continuous monitoring of suspended sediment in an agricultural watershed. For summer periods the prevalence of clockwise hysteresis (74.1% of measured rainstorm events with SSC &gt, 25 mg L&minus, 1) appeared related to rainstorm behaviors.

Riparian buffers are important features that help to mitigate suspended sediment loads within rivers running through agricultural landscapes. Evaluating their effectiveness for sediment control by different modelling approaches can help direct beneficial management practices. The Soil and Water Assessment Tool (SWAT) model and the Adapted Neuro-Fuzzy Inference System (ANFIS) based model were used for prediction of suspended sediment concentrations (SSC) and sediment loads in the Mill River watershed (PEI, Canada). Those models were then used to assess the impact of riparian buffer widths in reducing sediment loads. The ANFIS model predicted measured SSC more accurately than the SWAT model. The relationship between buffer width and sediment reduction was asymptotic, and the relationship begins to plateau when the width reaches 50 m. Increasing the buffer width from 15 to 100 m led to an increase in sediment loads retention of 30.5% and 36.2% of the total stream sediment load for the SWAT and ANFIS models, respectively. This study highlighted that a data-driven ANFIS based model can be used to simulate the impact of land use changes on the sediment delivery in a river.

We evaluated passage of Osmerus mordax (Rainbow Smelt), Salvelinus fontinalis (Brook Trout), and Alosa pseudoharengus (Alewife) at 4 fishways on Prince Edward Island, Canada. We implanted fish with passive integrated transponder (PIT) tags and recorded their movements using stationary PIT antennas. Passage at different fishway types was highly variable. Nature-like fishways passed over 96% of Brook Trout and a maximum of 41.8% of Rainbow Smelt. The latter percentage is the highest yet reported in the literature for Rainbow Smelt. Pool-and-weir fishways passed nearly 100% of Brook Trout, 13.2% of Rainbow Smelt, and only 5.6% of Alewife. After passing a fishway, upstream residency was highly variable both among species as well as among individuals within species. Rainbow Smelt were observed spawning within nature-like fishways, suggesting that this type of fishway may also provide spawning habitat. Results highlight the challenge of passing non-salmonid species and indicate nature-like fishways are more appropriate than technical designs for passing Rainbow Smelt.

The organophosphate pesticide chlorpyrifos has been introduced to the marine environment via adsorption to agricultural soil runoff or as spray drift. Chlorpyrifos affects the survival of some larval decapod crustaceans, but no data exist on the impacts to the American lobster, Homarus americanus. The purpose of the present study was to assess the levels at which chlorpyrifos affects the survival of postlarval H. americanus. Using acute saltwater exposures, the 24- and 48-h median lethal concentrations were established for stage IV H. americanus (1.56 and 1.33 µg/L, respectively). Movement, acetylcholinesterase activity, intermoult period, specific growth rate, and moult increment were measured during exposure to sublethal concentrations. Movement patterns were assessed to establish a 48-h median inhibition concentration for cessation of normal movement (0.66 µg/L). Acetylcholinesterase activity was found to be inhibited immediately post-exposure to 0.50, 0.57, and 0.82 µg/L chlorpyrifos but could be recovered within a period (9-15 d) in clean seawater. Sublethal growth effects of increased intermoult period, decreased specific growth rate, and decreased moult increment were observed during exposure to an environmentally relevant concentration (0.82 µg/L). The present study suggests that H. americanus stage IV larvae were marginally less sensitive to chlorpyrifos compared with other decapods and that acute lethality of H. americanus postlarvae is not likely to occur with chlorpyrifos concentrations previously reported from aquatic environments. Environ Toxicol Chem 2019;38:1294-1301. © 2019 SETAC.

Summary Excessive nitrate loads from intensive potato production have been linked to the reoccurring anoxic events in many estuaries in Prince Edward Island (PEI), Canada. Community-led watershed-based nutrient reduction planning has been promoted as a strategy for water quality restoration and initial nitrate load criteria have been proposed for the impacted estuaries. An integrated modeling approach was developed to predict base flow nitrate loads to inform the planning activities in the groundwater-dominated agricultural watersheds. Nitrate load is calculated as base flow multiplied by the average of nitrate concentration at the receiving watershed outlet. The average of nitrate concentration is estimated as the integration of nitrate leaching concentration over the watershed area minus a nitrate loss coefficient that accounts for long-term nitrate storage in the aquifer and losses from the recharge to the discharge zones. Nitrate leaching concentrations from potato rotation systems were estimated with a LEACHN model and the land use areas were determined from satellite image data (2006–2009) using GIS. The simulated average nitrate concentrations are compared with the arithmetic average of nitrate concentration measurements in each of the 27 watersheds for model calibration and in 138 watersheds for model verifications during 2006–2009. Sensitivity of the model to the variations of land use mapping errors, nitrate leaching concentrations from key sources, and nitrate loss coefficient was tested. The calibration and verification statistics and sensitivity analysis show that the model can provide accurate nitrate concentration predictions for watersheds with drainage areas more than 5 km 2 and nitrate concentration over 2 mg N L −1 , while the model resolution for watersheds with drainage areas below 5 km 2 and/or nitrate concentration below 2 mg N L −1 may not be sufficient for nitrate load management purposes. Comparisons of normalized daily stream discharges among the active hydrometric stations indicated that stream base flow could be prorated for nitrate load calculation from the nearest gauging station in the absence of discharge measurements. Annual nitrate losses, including aquifer long-term storage, denitrification, and riparian plant uptake were estimated to be 0.8 mg N L −1 , corresponding to 3.4 kg N ha −1 . The maximum and median base flow nitrate loads to the estuaries from among the 27 calibration watersheds were predicted to be 28.4 and 8.7 kg N ha −1 respectively with a root mean square error (RMSE) as 2.3 kg N ha −1 . From among the 75 watersheds selected for model verification, the maximum and median base flow nitrate loads to the estuaries were estimated to be 29 and 5.5 kg N ha −1 respectively with RMSE as 2.6 kg N ha −1 . At the estuaries with nitrate loads above the medians, the predominant nitrate sources (75–98%) were derived from the potato rotation lands, highlighting the importance of N use management with potato production for water quality restoration; nitrate load derived from atmospheric N deposits was estimated to account for 3.6–13% of annual nitrate loads in watersheds with nitrate loads exceeding the median values. The application of the model to nutrient reduction planning in the Southwest River watershed implies that a significant change on cropping practices has to be made in order to mitigate the anoxic events in this highly impacted agricultural watershed.

Zinc and calcium have highly interwoven functions that are essential for cellular homeostasis. Here we first present a novel real-time flow cytometric technique to measure mitochondrial redox state and show it is modulated by zinc and calcium, individually and combined. We then assess the interactions of zinc and calcium on mitochondrial H2O2 production, membrane potential (ΔΨm), morphological status, oxidative phosphorylation (OXPHOS), complex I activity, and structural integrity. Whereas zinc at low doses and both cations at high doses individually and combined promoted H2O2 production, the two cations individually did not alter mitochondrial redox state. However, when combined at low and high doses the two cations synergistically suppressed and promoted, respectively, mitochondrial shift to a more oxidized state. Surprisingly, the antioxidants vitamin E and N-acetylcysteine showed pro-oxidant activity at low doses, whereas at high antioxidant doses NAC inhibited OXPHOS and dyscoupled mitochondria. Individually, zinc was more potent than calcium in inhibiting OXPHOS, whereas calcium more potently dissipated the ΔΨm and altered mitochondrial volume and ultrastructure. The two cations synergistically inhibited OXPHOS but antagonistically dissipated ΔΨm and altered mitochondrial volume and morphology. Overall, our study highlights the importance of zinc and calcium in mitochondrial redox regulation and functional integrity. Importantly, we uncovered previously unrecognized bidirectional interactions of zinc and calcium that reveal distinctive foci for modulating mitochondrial function in normal and disease states because they are potentially protective or damaging depending on conditions.

Oil sands-influenced process waters have been observed to cause reproductive effects and to induced CYP1A activity in fishes; however, little progress has been made in determining causative agents. Naphthenic acids (NAs) are the predominant organic compounds in process-affected waters, but due to the complexity of the mixture, it has been difficult to examine causal linkages in fishes. The aim of this study was to use in vitro assays specific to reproductive and CYP1A mechanisms to determine if specific acid extractable fractions of NAs obtained from oil sands-influenced water are active toward reproductive processes or interact with the Ah receptor responsible for CYP1A activity. NAs were extracted from aged oil sands-influenced waters by use of acid precipitation, and the mixture was fractionated into three acidic and one neutral fraction. The four fractions were examined for Ah receptor-mediated potency by use of the H4IIE-luc bioassay, effects on production of steroid hormones by use of the H295R steroidogenesis assay, and sex steroid receptor binding activity using the yeast estrogen screen and yeast androgen screen. The mixtures were characterized by high resolution mass spectrometry, (1)H nuclear magnetic resonance, and attenuated total reflectance infrared spectroscopy. The neutral fraction elicited Ah-receptor mediated activity after 24 h but not after 48 or 72 h. None of the fractions contained measurable levels of estrogen or androgen receptor agonists nor did they cause reductions in steroidogenesis. A number of fractions showed antiestrogenic or antiandrogenicity potency, with the neutral and main acidic fractions being the most potent. Neutral aromatic compounds are likely responsible for the CYP1A activity observed. Direct estrogenic, androgenic, or steroidogenic mechanisms are unlikely for NAs based on these results, but NAs act as potent antiandrogen or antiestrogens.

To investigate impacts of proposed oil sands aquatic reclamation techniques on benthic fish, white sucker (Catostomus commersonii Lacepede, 1803) were stocked in 2 experimental ponds-Demonstration Pond, containing aged fine tailings capped with fresh water, consistent with proposed end-pit lake designs, and South Bison Pond, containing aged unextracted oil sands material-to examine the effects of unmodified hydrocarbons. White sucker were stocked from a nearby reservoir at both sites in May 2010 and sampled 4 mo later to measure indicators of energy storage and utilization. Comparisons were then made with the source population and 2 reference lakes in the region. After exposure to aged tailings, white sucker had smaller testes and ovaries and reduced growth compared with the source population. Fish introduced to aged unextracted oil sands material showed an increase in growth over the same period. Limited available energy, endocrine disruption, and chronic stress likely contributed to the effects observed, corresponding to elevated concentrations of naphthenic acids, aromatic compounds in bile, and increased CYP1A activity. Because of the chemical and biological complexity of these systems, direct cause-effect relationships could not be identified; however, effects were associated with naphthenic acids, polycyclic aromatic hydrocarbons, ammonia, and high pH. Impacts on growth have not been previously observed in pelagic fishes examined in these systems, and may be related to differences in sediment interaction.

Oil sands-affected water from mining must eventually be incorporated into the reclaimed landscape or treated and released. However, this material contains petrogenic organic compounds, such as naphthenic acids and traces of polycyclic aromatic hydrocarbons. This has raised concerns for impacts of oil sands process-affected waters on the heath of wildlife and humans downstream of receiving environments. The objective of this study was to evaluate the temporal association of disease states in fish with water chemistry of oil sands-affected waters over more than a decade and determine the pathogens associated with disease pathologies. Yellow perch (Perca flavescens) captured from nearby lakes were stocked into two experimental ponds during 1995-1997 and 2008-2010. South Bison Pond is a drainage basin that has received unextracted oil sands-contaminated material. Demonstration Pond is a constructed pond containing mature fine tailings capped with fresh water. Two disease pathologies, fin erosion for which a suspected bacterial pathogen (Acinetobacter Iwoffi) is identified, and lymphocystis (confirmed using a real-time PCR) were associated with oil sands-affected water exposure. From 1995 to 1997 pathologies were most prevalent in the South Bison Pond; however, from 2008 to 2009, disease was more frequently observed in the Demonstration Pond. CYP1A activity was 3-16 fold higher in fish from experimental ponds as compared to reference populations and this pattern was consistent across all sampling years. Bile fluorescence displayed a gradient of exposure with experimental ponds being elevated over local perch populations. Naphthenic acids decreased in the Bison Pond from approximately 12 mg/L to4 mg/L while naphthenic acids increased in the Demonstration Pond from 6 mg/L to 12 mg/L due to tailings densification. Temporal changes in naphthenic acid levels, CYP1A activity and bile fluorescent metabolites correlate positively with incidence of disease pathologies whereas all inorganic water quality changes (major ions, pH, metals) were not associated with disease responses.

Eutrophic aquatic habitats are characterized by the proliferation of vegetation leading to a large standing biomass that upon decomposition may create hypoxic (low-oxygen) conditions. This is indeed the case in nutrient impacted estuaries of Prince Edward Island, Canada, where macroalgae, from the genusUlva,form submerged ephemeral mats. Hydrological forces and gases released from photosynthesis and decomposition lead to these mats occasionally floating to the water’s surface, henceforth termed floating mats. Here, we explore the hypothesis that floating mats are refugia during periods of sustained hypoxia/anoxia and examine how the invertebrate community responds to it. Floating mats were not always present, so in the first year (2013) sampling was attempted monthly and limited to when both floating and submerged mats occurred. In the subsequent year sampling was weekly, but at only one estuary due to logistical constraints from increased sampling frequency, and was not limited to when both mat types occurred. Water temperature, salinity, and pH were monitored bi-weekly with dissolved oxygen concentration measured hourly. The floating and submerged assemblages shared many of the same taxa but were statistically distinct communities; submerged mats tended to have a greater proportion of benthic animals and floating mats had more mobile invertebrates and insects. In 2014, sampling happened to occur in the weeks before the onset of anoxia, during 113 consecutive hours of sustained anoxia, and for four weeks after normoxic conditions returned. The invertebrate community on floating mats appeared to be unaffected by anoxia, indicating that these mats may be refugia during times of oxygen stress. Conversely, there was a dramatic decrease in animal abundances that remained depressed on submerged mats for two weeks. Cluster analysis revealed that the submerged mat communities from before the onset of anoxia and four weeks after anoxia were highly similar to each other, indicating recovery. When mobile animals were considered alone, there was an exponential relationship between the percentage of animals on floating mats, relative to the total number on both mat types, and hypoxia. The occupation of floating mats by invertebrates at all times, and their dominance there during hypoxia/anoxia, provides support for the hypothesis that floating mats are refugia.

This study builds upon the work of a multiagency consortium tasked with determining cost-effective solutions for the effects of pulp mill effluents on fish reproduction. A laboratory fathead minnow egg production test and chemical characterization tools were used to benchmark 81 effluents from 20 mills across Canada, representing the major pulping, bleaching, and effluent treatment technologies. For Kraft and mechanical pulp mills, effluents containing less than 20 mg/L BOD5 were found to have the greatest probability of having no effects. Organic loading, expressed as the total detected solvent-extractable components by gas chromatography/mass spectrometry (GC/MS), also correlated with decreased egg laying. Exceptions were found for specific Kraft, mechanical, and sulfite mills, suggesting yet unidentified causative agents are involved. Recycled fiber mill effluents, tested for the first time, were found to have little potential for reproductive effects despite large variations in BOD5 and GC/MS profiles...

Endocrine-disrupting chemicals are exogenous substances that can impact the reproduction of fish, potentially by altering circulating concentrations of 17β-estradiol (E2), testosterone (T), and 11-ketotestosterone (11-KT). Common methods to measure steroids in plasma samples include radioimmunoassays (RIAs) and enzyme-linked immunosorbant assays (ELISAs). The present study examines variability in E2, T, and 11-KT across 8 laboratories measuring reference and pulp mill effluent-exposed white sucker (Catostomus commersoni) plasma. We examine the contribution of assay type (RIA vs ELISA), standardized hormone extraction, location of values on the standard curve (upper and lower limits), and other variables on the ability to distinguish hormone levels between reference and exposed fish and the impact of these variables on quantitation of hormones in different laboratories. Of the 8 participating laboratories, 7 of 8 and 7 of 7 identified differences between sites for female E2 and female T, respectively, and 7 of 7 and 4 of 5 identified no differences between male T and male 11-KT. Notably, however, the ng/mL concentration of steroids measured across laboratories varied by factors of 10-, 6-, 14-, and 10-fold, respectively. Within laboratory intra-assay variability was generally acceptable and below 15%. Factors contributing to interlaboratory variability included calculation errors, assay type, and methodology. Based on the interlaboratory variability detected, we provide guidelines and recommendations to improve the accuracy and precision of steroid measurements in fish ecotoxicology studies. Environ Toxicol Chem 2014;33:847–857. © 2013 SETAC

Estuarine eutrophication as a result of agricultural land use, including the use of chemical fertilizers, is increasing worldwide. Prince Edward Island (PEI), Canada has very high agricultural intensity by international standards with approximately 44% of the land area under production, and some watersheds in excess of 75% agricultural land-use. The type of agriculture is also intensive with primarily row crops that have high chemical fertilizer and pesticide usage. In light of these stressors, the hypothesis of this study was that mummichog (Fundulus heteroclitus) population parameters would change with point and nonpoint source pollution, and that multivariate statistics could be used to draw associations with specific stressors. Fish were sampled on a monthly basis from May through August at 7 estuaries spanning a range of land use, nutrient, and contaminant loadings. A suite of environmental variables were simplified into 3 principal components: PC1 representing agricultural land use, N loading, and plant habitat, PC2 being dominated by sediment sand and silt distribution, and PC3 largely reflecting P loading and sediment organic matter. There were significant differences in abundance of both adult and young-of-the-year mummichog, and these changes associated most strongly with PC1, the largely N-driven agricultural influences. In contrast, somatic variables such as liver and gonad size did not show strong association with the environmental quality principal component scores. The sand and silt PC2 appeared to have the opposite association with the biological data, with siltier environments correlating to older, larger, less dense populations of mummichog. Although pesticide residues were detected in estuarine sediment, there was no clear relationship between these and watershed agricultural intensity or biochemical indicators. There was, however, a strong relationship between agricultural environmental variables (PC1) and in vitro steroid production that is suggestive of a potential chemical effect. Eutrophication appeared to be a primary stressor affecting mummichog populations, as nutrient enrichment was associated with changes in habitat variables and these in turn were associated with high mummichog density. Thus, mummichog population demographics appear to have use as an indicator of adverse or worsening conditions in estuaries. We concluded that, based on the subset of environmental factors evaluated, the nonpoint–source inputs of sediments and nutrients exerted the greatest influence on mummichog populations in PEI estuaries. Integr Environ Assess Manag 2013;9:496–507. © 2013 SETAC

Historic deforestation has deprived many river systems of their natural wood loadings. To study the effects of the loss of wood from waterways, a field trial was conducted in three small forested streams in New Zealand. The objectives were to (i) examine differences in fish assemblages among wooded pools (where wood provided cover), open pools and riffles and (ii) measure the effects of wood removal on channel morphology and fish assemblages. In the first part of the study, no significant differences were found in total fish density among the three habitats. However, total fish biomass was significantly higher in wooded pools (64% of total fish biomass) compared with open pools and riffles. Mean density and biomass of banded kokopu (Galaxias fasciatus) and mean biomass of longfin eel (Anguilla dieffenbachii) were highest in wooded pools, whereas the density and biomass of bluegill bully (Gobiomorphus hubbsi) and torrentfish (Cheimarrichthys fosteri) were highest in riffles. In the second part of the study, wood was removed from a 200-m section (treatment) in each stream, significantly reducing pool area and increasing the proportion of channel area and length in riffles. At the habitat scale, banded kokopu and large longfin eel were the two species mostly affected by wood removal. At the reach scale, banded kokopu biomass was significantly lower in the treatment sections. Although wooded pools were a small portion of total habitat, they provided important habitat for two of New Zealand's larger native fish taxa.

Biological and chemical endpoints were measured in white sucker collected downstream of Athabasca oil sands developments (AB, Canada) and compared with those at Calling Lake (AB, Canada), a reference location upstream of the Athabasca oil sands deposit. Naphthenic acid concentrations were also measured at 14 sites in the Athabasca River watershed. Concentrations of naphthenic acids were elevated in tributaries adjacent to oil sands mining developments. Tributary naphthenic acid profiles were more similar to aged oil sands process water than samples from the Athabasca River, suggesting an influence of tailings in the tributaries. White sucker showed higher energy storage in the Athabasca River as indicated by significantly higher condition and liver size. White sucker were not investing that energy into reproductive effort as measured by gonad size and fecundity, which were significantly reduced relative to the reference location. White sucker showed increased exposure to polycyclic aromatic hydrocarbons as indicated by hepatic cytochrome P4501A (CYP1A) activity and fluorescent bile metabolites, as well as higher concentrations of naphthenic acids in bile. Cadmium, copper, nickel, and selenium were also elevated in white sucker liver tissue compared with the reference location. Based on the exposure profile and response pattern observed, effects on energy storage and utilization in white sucker from the Athabasca River most likely resulted from exposure to polycyclic aromatic hydrocarbons derived from petrogenic and pyrolytic sources. Environ Toxicol Chem 2017;36:2058-2067. © 2017 SETAC.

Increased agricultural land use leads to accelerated erosion and deposition of fine sediment in surface water. Monitoring of suspended sediment yields has proven challenging due to the spatial and temporal variability of sediment loading. Reliable sediment yield calculations depend on accurate monitoring of these highly episodic sediment loading events. This study aims to quantify precipitation-induced loading of suspended sediments on Prince Edward Island, Canada. Turbidity is considered to be a reasonably accurate proxy for suspended sediment data. In this study, turbidity was used to monitor suspended sediment concentration (SSC) and was measured for 2 years (December 2012-2014) in three subwatersheds with varying degrees of agricultural land use ranging from 10 to 69 %. Comparison of three turbidity meter calibration methods, two using suspended streambed sediment and one using automated sampling during rainfall events, revealed that the use of SSC samples constructed from streambed sediment was not an accurate replacement for water column sampling during rainfall events for calibration. Different particle size distributions in the three rivers produced significant impacts on the calibration methods demonstrating the need for river-specific calibration. Rainfall-induced sediment loading was significantly greater in the most agriculturally impacted site only when the load per rainfall event was corrected for runoff volume (total flow minus baseflow), flow increase intensity (the slope between the start of a runoff event and the peak of the hydrograph), and season. Monitoring turbidity, in combination with sediment modeling, may offer the best option for management purposes.

Rainbow trout were exposed in situ to oil sands-affected waters for 21 d, either with or without an immune stimulation using inactivated Aeromonas salmonicida. Three aquatic systems were utilized for the experiment: a pond containing oil sands tailings capped with approximately 3 m of natural surface water, a second pond where unextracted oil sands materials were deposited in the watershed, and a reservoir receiving Athabasca River water as a reference caging location. The three systems showed a gradient of oil sands-related compounds, most notably, total naphthenic acids were highest in the system containing tailings (13 mg/L), followed by the system influenced by unextracted oil sands (4 mg/L), followed by the reference cage location (1 mg/L). Biochemical and chemical measures of exposure in rainbow trout showed the same trend, with the tailings-influenced system having the highest hepatic EROD activity and elevated bile fluorescence measured at phenanthrene wavelengths. Trout caged in the tailings-influenced location had significantly fewer leukocytes and smaller spleens as compared to the reference fish, though liver size and condition factor were unaffected. Fish in the tailings-influenced waters also demonstrated increased fin erosion, indicative of opportunistic infection. The trout exposed to tailing-influenced waters also showed a significantly decreased ability to produce antibodies to the inactivated A. salmonicida. Given the complexity of the exposure conditions, exact causative agents could not be determined, however, naphthenic acids, polycyclic aromatic hydrocarbons and pH correlate with the immunotoxic effects while elevated salinity or metals seem unlikely causes.

Artificial fertilizers are contributing to the replacement of eelgrass (Zostera marina) by sea lettuce (Ulva lactuca) in estuaries of Prince Edward Island (PEI), Canada. In this study, we found that the nearshore fish community differed between areas dominated by these two vegetations within an estuary in every month sampled (April–August). Adult northern pipefish (Syngnathus fuscus), threespine stickleback (Gasterosteus aculeatus), blackspotted stickleback (Gasterosteus wheatlandi), and Atlantic silverside (Menidia menidia) were most strongly associated with eelgrass, while mummichog (Fundulus heteroclitus), ninespine stickleback (Pungitius pungitius), and American eel (Anguilla rostrata) were often more numerous in sea lettuce. Sea lettuce stations tended to have more young-of-the-year mummichog, fourspine stickleback (Apeltes quadracus), and Gasterosteus sp. than eelgrass stations but fewer young-of-the-year northern pipefish and Atlantic silverside. Fish richness and abundance were significantly lower in the sea lettuce than eelgrass habitat during August when benthic hypoxia occurred. We conclude that the loss of eelgrass from PEI estuaries will result in significant declines in fish biodiversity.

A long-term monitoring study was conducted on effluents from a bleached kraft pulp and paper mill located in Eastern Canada. The study was designed to gain insights into temporal effluent variability with respect to fish reproduction as it related to production upsets, mill restarts and conditions affecting biological treatment performance. Final effluent quality was monitored between February 2007 and May 2009 using biochemical and chemical oxygen demand, total suspended solids, resin and fatty acids, a gas chromatographic profiling index, and the presence of methyl substituted 2-cyclopentenones. Selected effluent samples were evaluated for effects on fish reproduction (egg production) using a shortened version of the adult fathead minnow reproductive test. The events relating to negative effects on fish reproduction were upsets of the pulping liquor recovery system resulting in black liquor losses, operational upsets of the hardwood line resulting in the loss of oxygen delignification filtrates, and conditions that reduced the performance of biological treatment (e.g., mill shutdown and low ambient temperatures). The reductions in egg production observed in fathead minnow were associated with biochemical oxygen demand values > 20 mg/L, GC profiling indices > 1.2 and the presence of methyl-substituted 2-cyclopentenones at concentrations > 100 μg/L. This study demonstrated the importance of both in-plant measures for controlling the loss of organics as well as the optimum operation of biological effluent treatment for eliminating effluent-related effects on fish reproduction (egg production) in the laboratory.