Comparative Effects of Embryonic Metformin Exposure on Wild and Laboratory-Spawned Fathead Minnow (Pimephales promelas) 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.

Age, body size, growth and dietary habits: What are the key factors driving individual variability in mercury of lacustrine fishes in northern temperate lakes?

Fish total mercury concentration ([THg]) has been linked to various fish attributes, but the relative importance of these attributes in accounting for among-individual variation in [THg] has not been thoroughly assessed. We compared the contributions of ontogeny (age, length), growth (growth rate, body condition), and food web position (δ13C, δ15N) to among-individual variability in [THg] within populations of seven common fishes from 141 north temperate lakes. Ontogenetic factors accounted for most variation in [THg]; age was a stronger and less variable predictor than length for most species. Adding both indices of growth and food web position to these models increased explained variation (R2) in [THg] by 6-25% among species. Fish [THg] at age increased with growth rate, while fish [THg] at length decreased with growth rate, and the effect of body condition was consistently negative. Trophic elevation (inferred from δ15N) was a stronger predictor than primary production source (inferred from δ13C) for piscivores but not benthivores. Fish [THg] increased with δ15N in all species but showed a more variable relationship with δ13C. Among-individual variation in [THg] is primarily related to age or size in most temperate freshwater fishes, and effects of growth rate and food web position need to be considered in the context of these ontogenetic drivers.

Trophodynamics of trace elements in marine organisms from cold and remote regions of southern hemisphere.

Trace metals bioaccumulate in aquatic organisms and some of them biomagnify through food webs, posing a threat to the organisms or their human consumers. Although the trophodynamics of many trace metals is well known in the northern hemisphere, much less is known about metals in aquatic food webs from cold and remote coastal zones of the southern hemisphere. To fill this gap, we investigated the trophodynamics of Al, Co, Cr, Li, Mo, Ni, Sr, and V, which were measured in marine macroinvertebrates and fishes from inshore and offshore locations in each of the Chilean Patagonia and the Antarctic Peninsula area. In Patagonia, there was biodilution of these metals across the whole food web, while biomagnification of Li and Ni was significantly found across the lower food web at the offshore site. In Antarctica, significant biodilution of Al, Li, Ni, Mo, Sr and V occurred through the whole food web for the inshore site, but no tendency (biodilution or biomagnification) was found (p > 0.05) across the organisms at lower trophic levels for the offshore site. Our data suggest that the geographic location and species influences the trophodynamics of these trace elements and expand our understanding of metal fate in remote locations of the southern hemisphere.

Effects of municipal wastewater effluents on the digestive gland microbiome of wild freshwater mussels (Lasmigona costata).

Gut microbial communities are vital for maintaining host health, and are sensitive to diet, environment, and chemical exposures. Wastewater treatment plants (WWTPs) release effluents containing antimicrobials, pharmaceuticals, and other contaminants that may negatively affect the gut microbiome of downstream organisms. This study investigated changes in the diversity and composition of the digestive gland microbiome of flutedshell mussels (Lasmigona costata) from upstream and downstream of two large (service >100,000) WWTPs. Mussel digestive gland microbiome was analyzed following the extraction, PCR amplification, and sequencing of bacterial DNA using the V3-V4 hypervariable regions of the 16 S rRNA gene. Bacterial alpha diversity decreased at sites downstream of the second WWTP and these sites were dissimilar in beta diversity from sites upstream and downstream of the first upstream WWTP. The microbiomes of mussels collected downstream of the first WWTP had increased relative abundances of Actinobacteria, Bacteroidetes, and Firmicutes, with a decrease in Cyanobacteria, compared to upstream mussels. Meanwhile, those collected downstream of the second WWTP increased in Proteobacteria and decreased in Actinobacteria, Bacteroidetes, and Tenericutes. Increased Proteobacteria has been linked to adverse effects in mammals, but their functions in mussels is currently unknown. Finally, effluent-derived bacteria were found in the microbiome of mussels downstream of both WWTPs but not in those from upstream. Overall, results show that the digestive gland microbiome of mussels collected upstream and downstream of WWTPs differed, which has implications for altered host health and the transport of WWTP-derived bacteria through aquatic ecosystems.

Forest management influences the effects of streamside wet areas on stream ecosystems.

Riparian zones contain areas of strong hydrological connectivity between land and stream, referred to as variable source areas (VSAs), and are considered biogeochemical control points. However, little is known about whether VSAs influence stream communities and whether this connectivity is affected by forest management. To address this, we used multiple biotic and abiotic indicators to (1) examine the influence of VSAs on riparian vegetation and stream ecosystems by comparing VSA and non-VSA reaches and (2) explore how forest management may affect the influence of VSAs on stream ecosystems. We detected some significant differences between VSA and non-VSA reaches in the riparian vegetation (greater understory and lower tree density) and stream ecosystem indicators (greater dissolved organic matter aromaticity, microbial biomass, peroxidase activity and collector-gatherer density, and lower dissolved organic carbon concentrations, algal biomass, and predatory macroinvertebrate density), which suggests that VSAs may create a more heterotrophic ecosystem locally. However, we show some evidence that forest management activities (specifically, road density) can alter the influence of VSAs and eliminate the differences observed at lower forest management intensities, and that the most hydrologically connected areas seem more sensitive to disturbance. Therefore, we suggest that the heterogeneity in hydrological connectivity along riparian zones should be considered when planning forest harvesting operations and road building (e.g., wider riparian buffers around VSAs).

Prevalence and Intensity of Salmincola edwardsii in Brook Trout in Northwest New Brunswick, Canada.

Parasites can compromise the health and fitness of individual fish, and it is important to generate baseline information that can then be used to document changes in the abundance and distribution of potentially pathogenic parasites. The ectoparasitic copepod Salmincola edwardsii was assessed with respect to prevalence (percentage of infected fish per site), infection intensity (number of parasites per infected fish), and attachment location on Brook Trout Salvelinus fontinalis in northwest New Brunswick, Canada. Ten sample sites were assessed, with six sites on two streams in the Quisibis River basin and four sites on three streams in the Restigouche River basin. Parasite species identity was supported by 100% sequence identity with S. edwardsii in a variable region within 28S rDNA. The prevalence of fish infected per site ranged from 19.0% to 79.6%, with an overall prevalence of 48.5 ± 19.1% (mean ± SD) per site. Mean infection intensity was 1.5 ± 0.9 copepods/fish (range = 1-7), with parasites almost exclusively surrounding the dorsal fin and/or adipose fin (97.6%). There was no influence of trout age-class on parasite prevalence. Some fish presented with fin erosion at the site of parasite attachment (12.5%), and 6.2% also presented with hyperplastic skin lesions where no parasites were observed, that could be misinterpreted as secondary bacterial or fungal infections. Skin and fin damage were significantly more common when fish were infected with three or more individual parasites. The pathogenic potential of this parasite makes its presence noteworthy as a risk to salmonids that are both recreationally and ecologically important.

Ecological Legacy of DDT Archived in Lake Sediments from Eastern Canada.

Historic forest management practices led to widespread aerial application of insecticides, such as dichlorodiphenyltrichloroethane (DDT), to North American conifer forests during ∼1950-1970. Lake basins thus may provide an important archive of inputs and aquatic responses to these organochlorines. We use dated sediment cores from five study lakes in multiple watersheds in New Brunswick (NB), Canada, to provide a regional paleo-ecotoxicological perspective on this potential legacy stressor in remote lake ecosystems. Peak sedimentary levels of p, p'- and o, p'-DDT (ΣDDT) and breakdown products ΣDDE (dichlorodiphenyldichloroethylene) and ΣDDD (dichlorodiphenyldichloroethane) generally occurred during the 1970s to 1980s. Sediments exceeded probable effect levels (PELs) by ∼450 times at the most impacted lake. Modern sediments in all study lakes still contained levels of DDT-related compounds that exceed PELs. For the first time, we show that aerial applications of DDT to eastern Canadian forests likely resulted in large shifts to primary consumers within several lake food webs, principally through lake-specific impacts on zooplankton community composition. Modern pelagic zooplankton communities are now much different compared to communities present before DDT use, suggesting that a regional organochlorine legacy may exist in the modern food webs of many remote NB lakes.

Global change-driven effects on dissolved organic matter composition: Implications for food webs of northern lakes.

Northern ecosystems are experiencing some of the most dramatic impacts of global change on Earth. Rising temperatures, hydrological intensification, changes in atmospheric acid deposition and associated acidification recovery, and changes in vegetative cover are resulting in fundamental changes in terrestrial-aquatic biogeochemical linkages. The effects of global change are readily observed in alterations in the supply of dissolved organic matter (DOM)-the messenger between terrestrial and lake ecosystems-with potentially profound effects on the structure and function of lakes. Northern terrestrial ecosystems contain substantial stores of organic matter and filter or funnel DOM, affecting the timing and magnitude of DOM delivery to surface waters. This terrestrial DOM is processed in streams, rivers, and lakes, ultimately shifting its composition, stoichiometry, and bioavailability. Here, we explore the potential consequences of these global change-driven effects for lake food webs at northern latitudes. Notably, we provide evidence that increased allochthonous DOM supply to lakes is overwhelming increased autochthonous DOM supply that potentially results from earlier ice-out and a longer growing season. Furthermore, we assess the potential implications of this shift for the nutritional quality of autotrophs in terms of their stoichiometry, fatty acid composition, toxin production, and methylmercury concentration, and therefore, contaminant transfer through the food web. We conclude that global change in northern regions leads not only to reduced primary productivity but also to nutritionally poorer lake food webs, with discernible consequences for the trophic web to fish and humans.

Is There a Risk to Humans from Consuming Octopus Species from Sites with High Environmental Levels of Metals?

Although octopuses are in high-demand globally and can bioaccumulate high concentrations of metals, the risk associated to its consumption is poorly understood. We compiled literature data from eight locations in Europe, North America and Northern Africa with different environmental levels of metals to evaluate: (1) the risk to human health through consumption of muscle tissues and digestive glands of Octopus hubbsorum, O. vulgaris and Eledone cirrhosa, and (2) the maximum allowable consumption rates (CRlim). The assessments were done according the United States Environmental Protection Agency. The results indicate the absence of health risk through consumption of muscle tissues, and high health risk through consumption of digestive glands (attributed to its high Cd levels), reflected in its 1ow CRlim values. In conclusion, even in contaminated sites the consumption of octopus muscle tissues is not an issue of concern, while the consumption of digestive glands should be more carefully monitored to avoid health problems.

Mercury bioaccumulation in aquatic biota along a salinity gradient in the Saint John River estuary.

Although estuaries are critical habitats for many aquatic species, the spatial trends of toxic methylmercury (MeHg) in biota from fresh to marine waters are poorly understood. Our objective was to determine if MeHg concentrations in biota changed along a salinity gradient in an estuary. Fourspine Stickleback (Apeltes quadracus), invertebrates (snails, amphipods, and chironomids), sediments, and water were collected from ten sites along the Saint John River estuary, New Brunswick, Canada in 2015 and 2016, with salinities ranging from 0.06 to 6.96. Total mercury (proxy for MeHg) was measured in whole fish and MeHg was measured in a subset of fish, pooled invertebrates, sediments, and water. Stable sulfur (δ34S), carbon (δ13C), and nitrogen (δ15N) isotope values were measured to assess energy sources (S, C) and relative trophic level (N). There were increases in biotic δ13C and δ34S from fresh to more saline sites and these measures were correlated with salinity. Though aqueous MeHg was higher at the freshwater than more saline sites, only chironomid MeHg increased significantly with salinity. In the Saint John River estuary, there was little evidence that MeHg and its associated risks increased along a salinity gradient.

Quantification of sulphur amino acids by ultra-high performance liquid chromatography in aquatic invertebrates.

We examined the performance of an ultra-high performance liquid chromatography method to quantify protein-bound sulphur amino acids in zooplankton. Both cysteic acid and methionine sulfone were linear from 5 to 250 pmol (r2 = 0.99), with a method detection limit of 13 pmol and 9 pmol, respectively. Although there was no matrix effect on linearity, adjacent peaks and co-eluting noise from the invertebrate proteins increased the detection limits when compared to common standards. Overall, performance characteristics were reproducible and accurate, and provide a means for quantifying sulphur amino acids in aquatic invertebrates, an understudied group.

A proposed framework for the systematic review and integrated assessment (SYRINA) of endocrine disrupting chemicals.

BACKGROUND: The issue of endocrine disrupting chemicals (EDCs) is receiving wide attention from both the scientific and regulatory communities. Recent analyses of the EDC literature have been criticized for failing to use transparent and objective approaches to draw conclusions about the strength of evidence linking EDC exposures to adverse health or environmental outcomes. Systematic review methodologies are ideal for addressing this issue as they provide transparent and consistent approaches to study selection and evaluation. Objective methods are needed for integrating the multiple streams of evidence (epidemiology, wildlife, laboratory animal, in vitro, and in silico data) that are relevant in assessing EDCs. METHODS: We have developed a framework for the systematic review and integrated assessment (SYRINA) of EDC studies. The framework was designed for use with the International Program on Chemical Safety (IPCS) and World Health Organization (WHO) definition of an EDC, which requires appraisal of evidence regarding 1) association between exposure and an adverse effect, 2) association between exposure and endocrine disrupting activity, and 3) a plausible link between the adverse effect and the endocrine disrupting activity. RESULTS: Building from existing methodologies for evaluating and synthesizing evidence, the SYRINA framework includes seven steps: 1) Formulate the problem; 2) Develop the review protocol; 3) Identify relevant evidence; 4) Evaluate evidence from individual studies; 5) Summarize and evaluate each stream of evidence; 6) Integrate evidence across all streams; 7) Draw conclusions, make recommendations, and evaluate uncertainties. The proposed method is tailored to the IPCS/WHO definition of an EDC but offers flexibility for use in the context of other definitions of EDCs. CONCLUSIONS: When using the SYRINA framework, the overall objective is to provide the evidence base needed to support decision making, including any action to avoid/minimise potential adverse effects of exposures. This framework allows for the evaluation and synthesis of evidence from multiple evidence streams. Finally, a decision regarding regulatory action is not only dependent on the strength of evidence, but also the consequences of action/inaction, e.g. limited or weak evidence may be sufficient to justify action if consequences are serious or irreversible.

The combined influence of two agricultural contaminants on natural communities of phytoplankton and zooplankton.

Concentrations of glyphosate observed in the environment are generally lower than those found to exert toxicity on aquatic organisms in the laboratory. Toxicity is often tested in the absence of other expected co-occurring contaminants. By examining changes in the phytoplankton and zooplankton communities of shallow, partitioned wetlands over a 5 month period, we assessed the potential for direct and indirect effects of the glyphosate-based herbicide, Roundup WeatherMax(©) applied at the maximum label rate, both in isolation and in a mixture with nutrients (from fertilizers). The co-application of herbicide and nutrients resulted in an immediate but transient decline in dietary quality of phytoplankton (8.3 % decline in edible carbon content/L) and zooplankton community similarity (27 % decline in similarity and loss of three taxa), whereas these effects were not evident in wetlands treated only with the herbicide. Thus, even at a worst-case exposure, this herbicide in isolation, did not produce the acutely toxic effects on plankton communities suggested by laboratory or mesocosm studies. Indirect effects of the herbicide-nutrient mixture were evident in mid-summer, when glyphosate residues were no longer detectable in surface water. Zooplankton abundance tripled, and zooplankton taxa richness increased by an average of four taxa in the herbicide and nutrient treated wetlands. The lack of significant toxicity of Roundup WeatherMax alone, as well as the observation of delayed interactive or indirect effects of the mixture of herbicide and nutrients attest to the value of manipulative field experiments as part of a comprehensive, tiered approach to risk assessments in ecotoxicology.

Recovery of a wild fish population from whole-lake additions of a synthetic estrogen.

Despite widespread recognition that municipal wastewaters contain natural and synthetic estrogens, which interfere with development and reproduction of fishes in freshwaters worldwide, there are limited data on the extent to which natural populations of fish can recover from exposure to these compounds. We conducted whole-lake additions of an active component of the birth control pill (17α-ethynylestradiol; EE2) that resulted in the collapse of the fathead minnow (Pimephales promelas) population. Here we quantify physiological, population, and genetic characteristics of this population over the 7 years after EE2 additions stopped to determine if complete recovery was possible. By 3 years post-treatment, whole-body vitellogenin concentrations in male fathead minnow had returned to baseline, and testicular abnormalities were absent. In the spring of the fourth year, adult size-frequency distribution and abundance had returned to pretreatment levels. Microsatellite analyses clearly showed that postrecovery fish were descendants of the original EE2-treated population. Results from this whole-lake experiment demonstrate that fish can recover from EE2 exposure at the biochemical through population levels, although the timelines to do so are long for multigenerational exposures. These results suggest that wastewater treatment facilities that reduce discharges of estrogens and their mimics can improve the health of resident fish populations in their receiving environments.

Perfluorinated and polyfluorinated compounds in lake food webs from the Canadian high Arctic.

Per- and polyfluorinated alkyl substances (PFASs) enter Arctic lakes through long-range atmospheric transport and local contamination, but their behavior in aquatic food webs at high latitudes is poorly understood. This study compared the concentrations of perfluorocarboxylates, perfluorosulfonates, and fluorotelomer sulfonates (FTS) in biotic and abiotic samples from six high Arctic lakes near Resolute Bay, Nunavut, Canada. Two of these lakes are known to be locally contaminated by a small airport and Arctic char (Salvelinus alpinus) from these lakes had over 100 times higher total [PFAS] when compared to fish from neighboring lakes. Perfluorononanoate (PFOA) and perfluorooctanesulfonate (PFOS) dominated in char, benthic chironomids (their main prey), and sediments, while pelagic zooplankton and water were dominated by lower chain acids and perfluorodecanesulfonate (PFDS). This study also provides the first measures of perfluoroethylcyclohexanesulfonate (PFECHS) and FTS compounds in water, sediment, juvenile char, and benthic invertebrates from lakes in the high Arctic. Negative relationships between [PFAS] and δ(15)N values (indicative of trophic position) within these food webs indicated no biomagnification. Overall, these results suggest that habitat use and local sources of contamination, but not trophic level, are important determinants of [PFAS] in biota from freshwater food webs in the Canadian Arctic.

Manufacturing doubt about endocrine disrupter science--A rebuttal of industry-sponsored critical comments on the UNEP/WHO report "State of the Science of Endocrine Disrupting Chemicals 2012".

We present a detailed response to the critique of "State of the Science of Endocrine Disrupting Chemicals 2012" (UNEP/WHO, 2013) by financial stakeholders, authored by Lamb et al. (2014). Lamb et al.'s claim that UNEP/WHO (2013) does not provide a balanced perspective on endocrine disruption is based on incomplete and misleading quoting of the report through omission of qualifying statements and inaccurate description of study objectives, results and conclusions. Lamb et al. define extremely narrow standards for synthesizing evidence which are then used to dismiss the UNEP/WHO 2013 report as flawed. We show that Lamb et al. misuse conceptual frameworks for assessing causality, especially the Bradford-Hill criteria, by ignoring the fundamental problems that exist with inferring causality from empirical observations. We conclude that Lamb et al.'s attempt of deconstructing the UNEP/WHO (2013) report is not particularly erudite and that their critique is not intended to be convincing to the scientific community, but to confuse the scientific data. Consequently, it promotes misinterpretation of the UNEP/WHO (2013) report by non-specialists, bureaucrats, politicians and other decision makers not intimately familiar with the topic of endocrine disruption and therefore susceptible to false generalizations of bias and subjectivity.

A path forward in the debate over health impacts of endocrine disrupting chemicals.

Several recent publications reflect debate on the issue of "endocrine disrupting chemicals" (EDCs), indicating that two seemingly mutually exclusive perspectives are being articulated separately and independently. Considering this, a group of scientists with expertise in basic science, medicine and risk assessment reviewed the various aspects of the debate to identify the most significant areas of dispute and to propose a path forward. We identified four areas of debate. The first is about the definitions for terms such as "endocrine disrupting chemical", "adverse effects", and "endocrine system". The second is focused on elements of hormone action including "potency", "endpoints", "timing", "dose" and "thresholds". The third addresses the information needed to establish sufficient evidence of harm. Finally, the fourth focuses on the need to develop and the characteristics of transparent, systematic methods to review the EDC literature. Herein we identify areas of general consensus and propose resolutions for these four areas that would allow the field to move beyond the current and, in our opinion, ineffective debate.

Environmental, geographic and trophic influences on methylmercury concentrations in macroinvertebrates from lakes and wetlands across Canada.

Macroinvertebrates are a key vector in the transfer of methylmercury (MeHg) to fish. However, the factors that affect MeHg concentrations and bioaccumulation in these organisms are not as well understood as for fish, and studies on a broad geographic scale are lacking. In this study, we gathered published and unpublished MeHg and carbon (δ(13)C) and nitrogen (δ(15)N) stable isotope data for freshwater macroinvertebrates from 119 lakes and wetlands across seven Canadian provinces, along with selected physical, chemical and biological characteristics of these systems. Overall, water pH was the most important determinant of MeHg concentrations in both predatory and non-predatory invertebrates [[Formula: see text] = 0.32, p < 0.001; multivariate canonical redundancy analysis (RDA)]. The location of lakes explained additional variation in invertebrate MeHg (partial R(2) = 0.08 and 0.06 for latitude and longitude, respectively; RDA), with higher concentrations in more easterly and southerly regions. Both invertebrate foraging behaviour and trophic position (indicated by functional feeding groups and δ(15)N values, respectively) also predicted MeHg concentrations in the organisms. Collectively, results indicate that in addition to their feeding ecology, invertebrates accumulate more MeHg in acidic systems where the supply of MeHg to the food web is typically high. MeHg concentrations in macroinvertebrates may also be influenced by larger-scale geographic differences in atmospheric mercury deposition among regions.