Effluent Exposure Research Articles

Battle of the Bites: The Effect of Sewage Effluent Exposure on Mosquitofish Biocontrol of Mosquitoes in Residential Louisiana.

Mosquitofish, Gambusia affinis, are eponymous larval mosquito predators. Their ability to colonize and survive in habitats that are uninhabitable by other potential predators allows them to naturally manage larval mosquito populations in most ground pools they are present in. However, effluent from residential onsite wastewater treatment systems (OWTSs) appears to limit the presence of fish predators. This is especially problematic in Louisiana, where regulations allow the discharge of OWTS effluent into open drainage conveyances. To determine the effect of effluent on the capacity of mosquitofish for biocontrol in contaminated areas, we assessed the body condition metrics of populations from two effluent-exposed sites and two sites not exposed to effluent, determined the lethal effect of effluent-contaminated drainage water on fish, and measured the prey consumption rates in the presence of effluent. Female fish collected from effluent-impacted sites had a reduced somatic body condition and most females examined displayed masculinized anal fins resembling the male gonopodium structure. This trait was not seen in fish collected from the control sites and has not yet been documented in association with OWTSs or in the state of Louisiana. Fish from the control sites survived at effluent-contaminated water levels < 70%, and the prey clearance rates increased with dilution. Onsite wastewater treatment system effluent has significant effects on both the short- and long-term persistence of mosquitofish, their body composition, reproductive health, and larval mosquito consumption. These effects likely release mosquito larvae from suppression and may increase the threat of mosquito-transmitted pathogens in effluent-contaminated locations.

The Effects of Wastewater Reuse on Smallmouth Bass (Micropterus dolomieu) Relative Abundance in the Shenandoah River Watershed, USA.

Municipal and industrial wastewater effluent is an important source of water for lotic systems, especially during periods of low flow. The accumulated wastewater effluent flows-expressed as a percentage of total streamflow (ACCWW%)-contain chemical mixtures that pose a risk to aquatic life; fish may be particularly vulnerable when chronically exposed. Although there has been considerable focus on individual-level effects of exposure to chemical mixtures found in wastewater effluent, scaling up to population-level effects remains a challenging component needed to better understand the potential consequences of exposure in wild populations. This may be particularly important under a changing climate in which wastewater reuse could be essential to maintain river flows. We evaluated the effects of chronic exposure to wastewater effluent, as measured by ACCWW%, on the relative abundance of young-of-year (YOY), juvenile, and adult smallmouth bass (Micropterus dolomieu) populations in the Shenandoah River Watershed (USA). We found that increases in ACCWW% in the previous year and during the prespawn period were negatively correlated with the relative abundance of YOY, resulting in an average 41% predicted decrease in abundance (range = 0.5%-94% predicted decrease in abundance). This lagged effect suggests that adult fish reproductive performance may be compromised by chemical exposure during periods of high ACCWW%. No relationships between ACCWW% and juvenile or adult relative abundance were found, suggesting that negative effects of ACCWW% on YOY abundance may be offset due to compensatory mechanisms following higher ACCWW% exposure. Understanding the effects of wastewater effluent exposure at multiple levels of biological organization will help in the development of management strategies aimed at protecting aquatic life. Environ Toxicol Chem 2024;43:1138-1148. © 2024 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.

Effects of diluted effluent on aquatic macroinvertebrate communities at the McClean Lake uranium operation in northern Saskatchewan

Diluted treated effluent from the McClean Lake uranium mill in northern Saskatchewan is released into Vulture Lake, which flows into the east basin of McClean Lake; this input could potentially cause a variety of disturbances to the aquatic systems. This study aimed to determine the potential effects of diluted effluent exposure (metals and major ions) on benthic macroinvertebrates in Vulture Lake and McClean Lake. Two monitoring locations located in Vulture Lake and eight in McClean Lake were used for collection water, sediment, and benthic macroinvertebrates. Complementary surface water bioassays were performed with larvae of the midge Chironomus dilutus using lake water from selected sites. Results indicated that total macroinvertebrate abundance and Margalef index (MI) did not follow the diluted effluent pattern. In addition, while the MI from artificial substrate samplers showed higher values in Vulture Lake and lower values at McClean Lake sites 4 and 5 (closer to effluent diffuser), the values recorded for sediment grab samples registered lower indices in Vulture Lake and higher values for sites 4 and 5. The final model from a Generalized Additive Modelling (GAM) approach suggested that electrical conductivity (EC), selenium (Se), and chloride (Cl) in water, and total organic carbon (TOC) and cadmium (Cd) in sediment are key variables that collectively may have influenced macroinvertebrate community composition at the study sites. Finally, across all test endpoints in the bioassays, exposure to lake water from Vulture Lake and McClean Lake had no statistically significant effects on C. dilutus.

Evaluation of cyto-genotoxicity biomarkers, changes in histology and antioxidant defense system of Oreochromis niloticus induced by the industrial effluents

Aquatic pollution mainly by industrial effluents has been a major concern since a few decades. The current study evaluated cyto-genotoxicity of industrial effluents on Oreochromis niloticus exposed to sublethal levels by hematotoxicity, blood biochemistry analysis, micronucleus assay, antioxidants and cerebral toxicity. The significant elevation in differential leukocytes of exposed fish was indicative of infections and compromised immune system. The acute and chronic industrial effluent exposure caused significant decline in aspartame transaminase (AST) and alanine transaminase (ALT) and renal function enzymes. Necrosis, hyperplastic growth, hypertrophy and toxicant accumulation exhibited cerebral toxicity potential of industrial toxicants. A significant decrease in antioxidants, GSH, SOD and catalase (0.14, 0.66 and 1549 unit/mg protein) in chronic exposure group in comparison to 0.18, 2.83, 7680 and 6200.8 values of GSH, SOD, GPx and CAT, respectively. Results showed that acute and chronic industrial effluent exposure caused genotoxicity with higher frequencies of formation of micronuclei and cytokaryotic fusion.

Toxicological approaches as tool to assess the effects of a mixture of photocatalytic degradation products originated from the unregulated neonicotinoid acetamiprid employing a terrestrial organism (Eisenia andrei)

Acetamiprid (ACT) has been detected in several water sources in Latin America. The presence of its degradation products in the environment is not negligible and transformation products (TPs) significantly contribute to environmental health risks. Although advanced oxidative processes are promising for the treatment of this neocotinoid, effects of these are still unknown. In this context, the effects of a mixture of photocatalytic degradation products resulting from an ACT treatment for 90 min employing TiO2/UV on cytotoxicity and oxidative stress parameters in Eisenia andrei earthworms in acute and chronic experiments using typical Latin American soil were assessed. Acute contact tests were performed (72 h) using a filter paper moistened with an ACT solution and a chronic test was performed using Oxisoil (200 g) moistened with an ACT solution for 45 days. Catalase (CAT) and glutathione-S-transferase (GST) activities, reduced glutathione (GSH) levels and cytotoxicity (cellular eleocyte and amoebocyte assessments) were investigated. Over 75 % of ACT was degraded within the first 15 min of treatment, with levels below the limit of detection after 60 min. The acute test revealed greater cytotoxic effects associated with the effluents treated for T0 and T15 min, with decreased cell density noted after 48 h of exposure, in addition to CAT induction (in all treatments) and GST induction following T0, T15 and T90 min exposures. Concerning the chronic assay, decreases in cell density (T0, T15, T60 and T90 min) and viability (T0, T60 and T90 min) were observed after 45 days, in addition to induced CAT activity following T0, T15 and T60 exposures and GST induction following the T60 min exposure. Reduced glutathione levels were unaltered, comprising the least sensitive biomarker among the investigated parameters to the treated effluent exposures. The mixture of ACT degradation products can cause toxic effects to non-target organisms, despite parent compound degradation, alerting for the need for ecotoxicological tests to prove decreased effluent toxicity, in addition to the improvement of degradation techniques.

Dysbiosis of gut microbiota by actual long-term exposure to textile wastewater treatment plant effluents in adult zebrafish (Danio rerio) using aquatic microcosm systems

In the future, textile industry wastewater may create greater possibilities of releasing emerging contaminants into aquatic environments. Research on textile dyeing wastewater has focused on its adverse effects on biota. However, the effects of textile wastewater on gut microbiome and host health have not yet been elucidated, especially for the toxic assessment of combined pollutants under receiving water environment conditions. During this study, adult zebrafish were exposed to realistic textile wastewater treatment plant (TWTP) effluents in aquatic microcosm systems (AMS) for six months, a histopathological examination was performed, after which 16S rRNA and metagenomic sequencing of the intestinal microbiota was conducted and fish growth performance was also examined. As a result of exposure to TWTP effluents, adult zebrafish growth was significantly inhibited, the condition factor was exacerbated, and mortality was significantly increased. Also, significant alterations were observed in zebrafish guts after chronic exposure, which are the opportunistic pathogens Flavobacterium, Aeromonas, and Escherichia. The chronic effects of increased probiotic Cetobacterium, Bacteroides, and Planctomyces deserve further attention. Metagenomic sequencing also showed that genes of core metabolism pathways were affected by TWTP effluent exposure. The study has, for the first time, demonstrated gut microbiota's susceptibility and host health of adult zebrafish to TWTP effluents using aquatic microcosm systems, which are assumed to be clean, thus warranting additional research to reveal their mode of toxicity.

Sampling method and season influence selenium dynamics at the base of a boreal lake food chain

Few studies have investigated the potential influence of sampling method and season on Se bioaccumulation at the base of the aquatic food chain. In particular, the effects of low water temperature associated with prolonged ice-cover periods on Se uptake by periphyton and further transfer to benthic macroinvertebrates (BMI) have been overlooked. Such information is crucial to help improve Se modelling and risk assessment at sites receiving continuous Se inputs. To date, this seems to be the first study to address these research questions. Here, we examined potential differences related to sampling methods (artificial substrates vs. grab samples) and seasons (summer vs. winter) on Se dynamics in the benthic food chain of a boreal lake (McClean Lake) receiving continuous low-level Se input from a Saskatchewan uranium milling operation. During summer 2019, water, sediment grab samples and artificial substrates were sampled from 8 sites with varying mill-treated effluent exposure. In winter 2021, water and sediment grab samples were sampled at 4 locations in McClean Lake. Water, sediment, and biological samples were subsequently analyzed for total Se concentrations. Enrichment functions (EF) in periphyton and trophic transfer factors (TTF) in BMI were calculated for both sampling methods and seasons. Periphyton collected with artificial substrates (Hester-Dendy samplers and glass plates) exhibited significantly higher mean Se concentrations (2.4 ± 1.5 μg/g d.w) than periphyton collected from the surface of sediment grab samples (1.1 ± 1.3 μg/g d.w). Selenium concentrations in periphyton sampled in winter (3.5 ± 1.0 μg/g d.w) were significantly greater than summer (1.1 ± 1.3 μg/g d.w). Nevertheless, Se bioaccumulation in BMI was similar between seasons, possibly suggesting that invertebrates are not actively feeding in winter. Further investigations are necessary to verify if peak Se bioaccumulation in BMI takes place in spring, coinciding with the reproductive and developmental windows of some fish species.

Phytotoxicity and cytotoxicity attributes of immobilized Bacillus cereus treated and untreated textile effluents on Vigna mungo seeds and Artemia franciscana larvae

The physicochemical attributes of textile effluents collected from secondary treatment stage was investigated in this study and also assess the biosorption potential of membrane immobilized Bacillus cereus and free form of Bacillus cereus on textile effluent through bioreactor model study to find a sustainable solution to manage the textile effluent as vital need. Furthermore, the phytotoxicity and cytotoxicity nature of treated and untreated textile effluents on Vigna mungo and Artemia franciscana larvae under laboratory conditions as a novel approach. The textile effluent physicochemical parameter analysis results showed that the properties such as colour (Hazen unit), pH, turbidity, As, Biological Oxygen Demand (BOD), Chemical Oxygen Demand (COD), Cd, Cl, Cr, Cu, Hg, Ni, Pb, SO42−, and Zn were beyond the acceptable limits. Bacillus cereus immobilized on a polyethylene membrane eliminated greater amounts of dye (25.0 ± 1.3, 56.5 ± 1.8, 57.18 ± 1.5, and 54.34 ± 1.7 Hazen unit from An1, Ae2, Ve3, and So4 respectively) and pollutants (As: 0.9–2.0, Cd: 6–8, Cr: 300–450, Cu: 5–7, Hg: 0.1–0.7, Ni: 8–14, Pb: 4–5, and Zn: 4–8 mg L−1) from textile effluent in a week of biosorption investigation using a bioreactor model (batch type) compared to a free form of B. cereus on textile effluent. The phytotoxicity and cytotoxicity study results revealed that the membrane immobilized B. cereus treated textile effluent exposure showed reduced phytotoxicity and minimal cytotoxicity (including mortality) percentage compared with free form B. cereus treated and untreated textile effluents. These entire results conclude that the membrane immobilized B. cereus may considerably minimize/detoxify the harmful pollutants from the textile effluents. A large scale level biosorption approach need to be performed to validate the maximum pollutants removing potential of this membrane immobilized bacteria species and optimal conditions for effective remediation.

Transcriptome signatures of wastewater effluent exposure in larval zebrafish vary with seasonal mixture composition in an effluent-dominated stream

Wastewater treatment plant (WWTP) effluent-dominated streams provide critical habitat for aquatic and terrestrial organisms but also continually expose them to complex mixtures of pharmaceuticals that can potentially impair growth, behavior, and reproduction. Currently, few biomarkers are available that relate to pharmaceutical-specific mechanisms of action. In the experiment reported in this paper, zebrafish (Danio rerio) embryos at two developmental stages were exposed to water samples from three sampling sites (0.1 km upstream of the outfall, at the effluent outfall, and 0.1 km below the outfall) during base-flow conditions from two months (January and May) of a temperate-region effluent-dominated stream containing a complex mixture of pharmaceuticals and other contaminants of emerging concern. RNA-sequencing identified potential biological impacts and biomarkers of WWTP effluent exposure that extend past traditional markers of endocrine disruption. Transcriptomics revealed changes to a wide range of biological functions and pathways including cardiac, neurological, visual, metabolic, and signaling pathways. These transcriptomic changes varied by developmental stage and displayed sensitivity to variable chemical composition and concentration of effluent, thus indicating a need for stage-specific biomarkers. Some transcripts are known to be associated with genes related to pharmaceuticals that were present in the collected samples. Although traditional biomarkers of endocrine disruption were not enriched in either month, a high estrogenicity signal was detected upstream in May and implicates the presence of unidentified chemical inputs not captured by the targeted chemical analysis. This work reveals associations between bioeffects of exposure, stage of development, and the composition of chemical mixtures in effluent-dominated surface water. The work underscores the importance of measuring effects beyond the endocrine system when assessing the impact of bioactive chemicals in WWTP effluent and identifies a need for non-targeted chemical analysis when bioeffects are not explained by the targeted analysis.

Impact of Paper Mill Effluent on Carbohydrate Metabolism of Freshwater Snake Headed Fish, Channa punctatus (Bloch, 1793)

The snake-headed fish, Channa sp. Bloch (Actinoptrygii: Channidae) is animportant component of inland fishery in the Indian subcontinent and one of the fashionable commercial fish. The industrialized effluents’ accomplishment to the aquatic ecosystem is one of the foremost causes of environmental pollution, and its hazardous influence on the exposed edible fishes may be a serious threat to human health. The present investigation has been designed to study the effects of sublethal concentrations of paper mill effluent on the serum glucose, serum lactic acid, liver glycogen, and liver lactic acid contents of Channa punctatus (Bloch) after in vitro exposure for96 hours. The investigation recorded a significant increase in serum glucose, serum lactic acid, and liver lactic acid contents with a significant quantitative decline in liver glycogen among effluent exposed fishes were recorded during the investigation. Thus the authors supposed to propose that the carbohydrate metabolism of freshwater snake-headed fish, C. punctatus affected during effluent exposure based on their experimental findings.

Functional and phylogenetic responses of motile cryptofauna to habitat degradation.

Biodiversity of terrestrial and marine ecosystems, including coral reefs, is dominated by small, often cryptic, invertebrate taxa that play important roles in ecosystem structure and functioning. While cryptofauna community structure is determined by strong small-scale microhabitat associations, the extent to which ecological and environmental factors shape these communities are largely unknown, as is the relative importance of particular microhabitats in supporting reef trophodynamics from the bottom up. The goal of this study was to address these knowledge gaps, provided coral reefs are increasingly exposed to multiple disturbances and environmental gradients that influence habitat complexity, condition and ecosystem functioning. We compared the density, biomass, size range, phylogenetic diversity and functional roles of motile cryptofauna in Palau, Western Micronesia, among four coral-derived microhabitats representing various states of degradation (live coral [Acropora and Pocillopora], dead coral and coral rubble) from reefs along a gradient of effluent exposure. In total, 122 families across ten phyla were identified, dominated by the Arthropoda (Crustacea) and Mollusca. Cryptofauna biomass was greatest in live Pocillopora, while coral rubble contained the greatest density and diversity. Size ranges were broader in live corals than both dead coral and rubble. From a bottom-up perspective, effluent exposure had mixed effects on cryptic communities including a decline in total biomass in rubble. From a top-down perspective, cryptofauna were generally unaffected by predator biomass. Our data show that, as coral reef ecosystems continue to decline in response to more frequent and severe disturbances, habitats other than live coral may become increasingly important in supporting coral reef biodiversity and food webs.

Selenium Distribution and Trophic Transfer in the Periphyton-Benthic Macroinvertebrate Food Chain in Boreal Lakes Downstream from a Milling Operation.

Selenium (Se) is an essential micronutrient with a narrow essentiality-toxicity range known to bioaccumulate in aquatic food webs. Selenium uptake and trophic transfer at the base of aquatic food chains represent a great source of uncertainty for Se risk assessment. The goal of the present study was to investigate Se distribution in water and sediment and its subsequent transfer into the periphyton-benthic macroinvertebrate (BMI) food chain in boreal lakes downstream from a Saskatchewan uranium mill. In particular, the present study aimed to assess potential differences in Se bioaccumulation patterns by BMI taxa to contribute to the current knowledge gap. During summer 2018 and 2019, water, sediment, periphyton, and BMI were sampled at two sites in Vulture Lake, seven sites in McClean Lake east basin, and one reference site in McClean Lake west basin. Periphyton and BMI taxa were sampled with artificial substrates (Hester-Dendy) deployed for 5 weeks in 2018 and 7 weeks in 2019; BMI were sorted into the lowest practical achievable taxonomic level and analyzed for total Se concentrations. At the diluted effluent exposure sites, Se concentrations in BMI ranged from 1.3 to 18.0 µg/g dry weight and from 0.3 to 49.3 µg/g dry weight in 2018 and 2019, respectively, whereas concentrations ranged from 0.01 to 3.5 µg/g dry weight at the reference site. Selenium concentrations in periphyton and some BMI taxa sampled near the effluent diffuser (Se < 1 µg/L) reached levels comparable to higher effluent exposure sites (Se > 2 µg/L). Despite differences in Se bioaccumulation among BMI taxa, an approximately one-to-one trophic transfer ratio was observed for benthic primary consumers and benthic predatory taxa. Environ Toxicol Chem 2022;41:2181-2192. © 2022 SETAC.

Delineating Effluent Exposure and Cumulative Ecotoxicological Risk of Metals Downstream of a Saskatchewan Uranium Mill Using Autonomous Sensors.

There is increasing interest in using autonomous sensor technology to monitor aquatic ecosystems in real time and in employing such monitoring data to perform better ecological risk assessments. At seven locations in McClean Lake in northern Saskatchewan (Canada) that received diluted uranium milling effluent, we deployed sensor units to track effluent distribution and help predict potential biological effects on aquatic invertebrates. Water was also collected from each location on multiple occasions to measure major ions, dissolved metals, and routine water quality, and sediment was sampled to analyze total metals. The ecotoxicological risk to aquatic invertebrates was estimated using hazard quotients (HQs). The cumulative risk was estimated by summing the individual HQs, and the major ions risk was based on total osmolarity. The results indicated temporal and spatial variations in effluent exposure based on sensor electrical conductivity (EC) measurements in the McClean Lake East Basin. Individual HQs for water ranged from "moderate" (0.40-0.69) to "very high" (greater than 1) for silver, cadmium, arsenic, selenium, mercury, iron, and thallium. At all sites, major ions risk was less than 1. Individual HQs for sediment were "moderate" (0.40-0.69), "high" (0.7-0.99), and "very high" (greater than 1) for vanadium and cadmium. The cumulative risk in water and sediment for all metals combined was greater than 1 at some sites in Vulture Lake (which discharged into McClean Lake) and in McClean Lake itself. A more detailed estimation of the risks for aqueous selenium and arsenic (the only two metals that had good correlation with sensor EC data) indicated that their 90th percentile HQ values were less than 1 in McClean Lake, suggesting that these contaminants of concern do not represent a significant direct risk to aquatic invertebrate communities. Environ Toxicol Chem 2022;41:1765-1777. © 2022 SETAC.

Wastewater effluent affects behaviour and metabolomic endpoints in damselfly larvae

Wastewater treatment plant effluents have been identified as a major contributor to increasing anthropogenic pollution in aquatic environments worldwide. Yet, little is known about the potentially adverse effects of wastewater treatment plant effluent on aquatic invertebrates. In this study, we assessed effects of wastewater effluent on the behaviour and metabolic profiles of damselfly larvae (Coenagrion hastulatum), a common aquatic invertebrate species. Four key behavioural traits: activity, boldness, escape response, and foraging (traits all linked tightly to individual fitness) were studied in larvae before and after one week of exposure to a range of effluent dilutions (0, 50, 75, 100%). Effluent exposure reduced activity and foraging, but generated faster escape response. Metabolomic analyses via targeted and non-targeted mass spectrometry methods revealed that exposure caused significant changes to 14 individual compounds (4 amino acids, 3 carnitines, 3 lysolipids, 1 peptide, 2 sugar acids, 1 sugar). Taken together, these compound changes indicate an increase in protein metabolism and oxidative stress. Our findings illustrate that wastewater effluent can affect both behavioural and physiological traits of aquatic invertebrates, and as such might pose an even greater threat to aquatic ecosystems than previously assumed. More long-term studies are now needed evaluate if these changes are linked to adverse effects on fitness. The combination of behavioural and metabolomic assessments provide a promising tool for detecting effects of wastewater effluent, on multiple biological levels of organisation, in aquatic ecosystems.

Exposure duration and composition are important variables to predict short-term toxicity of effluents to a tropical copepod, Acartia sinjiensis

Predicting the toxicity of effluent exposures, which vary in duration, composition, and concentration, poses a challenge for ecological risk assessments. Effluent discharges may frequently result in the exposure of aquatic organisms to high concentrations of mixed contaminants for short durations. In the receiving environment effluents will undergo dilution and physical or chemical processes that further reduce contaminant concentrations at varying rates. To date, most studies comparing toxicity risks of continuous and pulsed contaminant exposures have focused on individual contaminants. In this study, the toxicity to the tropical euryhaline copepod Acartia sinjiensis of two complex effluents was assessed, comparing 6- and 18-h pulses and 78-h continuous exposures. Observations of larval development success and population size were completed after a 78-h incubation period, to observe for latent effects after pulse exposures. The chemical compositions of the effluents were assessed over time and different contaminants (i.e., metals, ammonia or organics) declined at differing rates. These were characterized as either a minimal, steady, or rapid decline. Nauplii development and population after 78 h were more impacted by effluent exposures following an 18-h pulse, compared to a 6-h pulse. Based on pulse-exposure concentrations, the 50% effect concentrations (EC50) were similar for continuous and 18-h exposures but up to 3-fold greater (lower toxicity) for the shorter 6-h exposures. Time-weighted average concentrations did not accurately predict toxicity from pulse exposures of the effluents. Concentration-addition toxicity modelling using toxicity data from pulse exposures of single contaminants was useful for predicting the toxicity of chemical mixtures exposed for varying durations. Recommendations for modified approaches to assessing risks of short-term effluent discharges are discussed.

Temperature modulates the impacts of wastewater exposure on the physiology and behaviour of fathead minnow

Municipal wastewater treatment plant (WWTP) effluent is a substantial source of pollution in aquatic habitats that can impact organisms across multiple levels of biological organization. Even though wastewater effluent is discharged continuously all year long, its impacts across seasons, specifically during winter, have largely been neglected in ecotoxicological research. Seasonal differences are of particular interest, as temperature-driven metabolic changes in aquatic organisms can significantly alter their ability to respond to chemical stressors. In this study, we examined the effects of multiple levels of wastewater effluent exposure (0, 25, or 50% treated effluent) on the physiological and behavioural responses of adult fathead minnow (Pimephales promelas) at temperatures simulating either summer (20 °C) or winter (4 °C) conditions. At 20 °C, wastewater exposure posed a metabolic cost to fish, demonstrated by higher standard metabolic rate and was associated with increased haematocrit and a reduction in boldness. In contrast, fish exposed to wastewater at 4 °C experienced no change in metabolic rate but performed fewer social interactions with their conspecifics. Taken together, our results demonstrate that wastewater exposure can lead to metabolic and behavioural disruptions, and such disruptions vary in magnitude and direction depending on temperature. Our findings highlight the importance of studying the interactions between stressors, while also underscoring the importance of research during colder periods of the year to broaden and deepen our understanding of the impacts of wastewater contamination in aquatic ecosystems.

RNA-seq reveals potential gene biomarkers in fathead minnows (Pimephales promelas) for exposure to treated wastewater effluent.

Discharged wastewater treatment plant (WWTP) effluent greatly contributes to the generation of complex mixtures of contaminants of emerging concern (CECs) in aquatic environments which often contain neuropharmaceuticals and other emerging contaminants that may impact neurological function. However, there is a paucity of knowledge on the neurological impacts of these exposures to aquatic organisms. In this study, caged fathead minnows (Pimephales promelas) were exposed in situ in a temperate-region effluent-dominated stream (i.e., Muddy Creek) in Coralville, Iowa, USA upstream and downstream of a WWTP effluent outfall. The pharmaceutical composition of Muddy Creek was recently characterized by our team and revealed many compounds there were at a low microgram to high nanogram per liter concentration. Total RNA sequencing analysis on brain tissues revealed 280 gene isoforms that were significantly differentially expressed in male fish and 293 gene isoforms in female fish between the upstream and downstream site. Only 66 (13%) of such gene isoforms overlapped amongst male and female fish, demonstrating sex-dependent impacts on neuronal gene expression. By using a systems biology approach paired with functional enrichment analyses, we identified several potential novel gene biomarkers for treated effluent exposure that could be used to expand monitoring of environmental effects with respect to complex CEC mixtures. Lastly, when comparing the results of this study to those that relied on a single-compound approach, there was relatively little overlap in terms of gene-specific effects. This discovery brings into question the application of single-compound exposures in accurately characterizing environmental risks of complex mixtures and for gene biomarker identification.

Metabolomics reveals changes in metabolite profiles due to growth and metamorphosis during the ontogeny of the northern damselfly

Many insects have complex life cycles where a drastic ontogenetic change happens between the larval stages and the adult stage, i.e. metamorphosis. Damselflies (order Odonata, suborder Zygoptera) are widely distributed and ecologically important semi-aquatic insects with a complex life cycle. Phenotypic changes over damselfly ontogeny have been documented, however, if and how metabolite profiles are also changing is currently unknown. Here we used a metabolomics methodology to gain insights into the metabolic changes during the life cycle of the Northern damselfly (Coenagrion hastulatum). Hatchlings of wild-caught damselflies were reared in the laboratory and metabolomics analyses using liquid chromatography and gas chromatography coupled to mass spectrometry were carried out at three larval stages and on adult damselflies. Additionally, a subset of larvae was exposed to wastewater effluent to assess how metabolite profiles responded to an environmental stressor. A total of 212 compounds belonging to several classes (e.g. amino acids, fatty acids, sugars) were annotated. Across metamorphosis, we found that damselflies shifted from protein catabolism to lipid catabolism. Wastewater effluent exposure resulted in ontogenetic stage-dependent changes of individual metabolites, but not to a marked extent. Overall, our study is one of the first to describe changes of metabolite profiles during ontogeny of an insect, and it provides a first step towards a greater understanding of the physiological changes occurring during general insect—but especially damselfly—ontogeny.

Effect of Chronic Exposure to Textile Wastewater Treatment Plant Effluents on Growth Performance, Oxidative Stress, and Intestinal Microbiota in Adult Zebrafish (Danio rerio)

The ever-increasing production and processing of textiles will lead to greater risks of releasing pollutants into the environment. Textile wastewater treatment plants (TWTPs) effluent are an important source of persistent toxic pollutants in receiving water bodies. The effects of specific pollutants on organisms are usually studied under laboratory conditions, and therefore, comprehensive results are not obtained regarding the chronic combined effects of pollutants under aquatic environmental conditions. Thus, this study aimed to determine the combined effects of TWTP effluents on the growth performance, oxidative stress, inflammatory response, and intestinal microbiota of adult zebrafish (Danio rerio). Exposure to TWTP effluents significantly inhibited growth, exacerbated the condition factor, and increased the mortality of adult zebrafish. Moreover, markedly decreases were observed in the activities of antioxidant enzymes, such as CAT, GSH, GSH-Px, MDA, SOD, and T-AOC, mostly in the intestine and muscle tissues of zebrafish after 1 and 4 months of exposure. In addition, the results demonstrated that TWTP effluent exposure affected the intestinal microbial community composition and decreased community diversity. Slight changes were found in the relative abundance of probiotic Lactobacillus, Akkermansia, and Lactococcus in zebrafish guts after chronic TWTP effluent exposure. The chronic toxic effects of slight increases in opportunistic pathogens, such as Mycoplasma, Stenotrophomonas, and Vibrio, deserve further attention. Our results reveal that TWTP effluent exposure poses potential health risks to aquatic organisms through growth inhibition, oxidative stress impairment of the intestine and muscles, and intestinal microbial community alterations.

Microbial risk assessment and mitigation options for wastewater treatment in Arctic Canada

Populations in Arctic Canada are strongly connected to, and draw sustenance from, the physical environment. Recreation and food harvesting locations, however, may be impacted by the basic wastewater treatment and disposal processes used in the region. Within these mixed socio-ecological systems, people may unknowingly be exposed to wastewater pathogens, either by direct contact or indirectly through activities resulting in exposure to contaminated locally harvested food. The objectives of this research are to estimate microbial health risks attributable to wastewater effluent exposure in Arctic Canada and evaluate potential mitigation options. A participatory quantitative microbial risk assessment (QMRA) approach was used. Specifically, community knowledge and information describing human activity patterns in wastewater-impacted environments was used with microbial water quality data to model a range of exposure scenarios and risk mitigation options. In several exposure scenario results, estimated individual annual risk of acute gastrointestinal illness exceeds a proposed tolerable target of 10−3. These scenarios include shore recreation and consumption of shellfish harvested near primary mechanical treatment plants at low tide, as well as travel in wetland portions of passive treatment sites during spring freshet. These results suggest that wastewater effluent exposures may be contributing to gastrointestinal illness in some Arctic communities. Mitigation strategies, including improved treatment and interventions aimed at deterring access to disposal areas reduce risk estimates across scenarios to varying degrees. Overall, well-designed passive systems appear to be the most effective wastewater treatment option for Arctic Canada in terms of limiting and managing associated microbial health risks. This research demonstrates a novel application of QMRA and provides science-based evidence to support public health, water, and sanitation decisions and investment in Arctic regions.