Developmental toxicity of pre-production plastic pellets affects a large swathe of invertebrate taxa.

Microplastics pose risks to marine organisms through ingestion, entanglement, and as carriers of toxic additives and environmental pollutants. Plastic pre-production pellet leachates have been shown to affect the development of sea urchins and, to some extent, mussels. The extent of those developmental effects on other animal phyla remains unknown. Here, we test the toxicity of environmental mixed nurdle samples and new PVC pellets for the embryonic development or asexual reproduction by regeneration of animals from all the major animal superphyla (Lophotrochozoa, Ecdysozoa, Deuterostomia and Cnidaria). Our results show diverse, concentration-dependent impacts in all the species sampled for new pellets, and for molluscs and deuterostomes for environmental samples. Embryo axial formation, cell specification and, specially, morphogenesis seem to be the main processes affected by plastic leachate exposure. Our study serves as a proof of principle for the potentially catastrophic effects that increasing plastic concentrations in the oceans and other ecosystems can have across animal populations from all major animal superphyla.

Neglected impacts of plant protection products on invertebrate aquatic biodiversity: a focus on eco-evolutionary processes.

The application of plant protection products (PPPs) may have delayed and long-term non-intentional impacts on aquatic invertebrates inhabiting agricultural landscapes. Such effects may induce population responses based on developmental and transgenerational plasticity, selection of genetic resistance, as well as increased extirpation risks associated with random genetic drift. While the current knowledge on such effects of PPPs is still scarce in non-target aquatic invertebrate species, evidences are accumulating that support the need for consideration of evolutionary components of the population response to PPPs in standard procedures of risk assessment. This mini-review, as part of a contribution to the collective scientific assessment on PPP impacts on biodiversity and ecosystem services performed in the period 2020-2022, presents a brief survey of the current results published on the subject, mainly in freshwater crustaceans, and proposes some research avenues and strategies that we feel relevant to fill this gap.

Review: Abandoned mines as a resource or liability for wildlife.

Abandoned Mine Lands (AMLs) are areas where previous mineral extraction or processing has occurred. Hundreds of thousands of AMLs exist within the United States. Contaminated runoff from AMLs can negatively affect the physiology and ecology of surrounding terrestrial and aquatic habitats and species and can be detrimental to human health. As a response, several U.S. federal and state agencies have launched programs to assess health risks associated with AMLs. In some cases, however, AMLs may be beneficial to specific wildlife taxa. There is a relative paucity of studies investigating the physiological and ecological impacts of AMLs on wildlife. We conducted a systematic review examining published scientific articles that assessed the negative and positive impacts of AMLs across invertebrate and vertebrate taxa. We also offer suggestions on evaluating AMLs to develop effective mitigation strategies that reduce their negative tole on human and wildlife communities. Peer-reviewed publications were screened across WebofScience, PubMed and Google Scholar databases. Abandoned mine lands were generally detrimental to wildlife, with adverse effects ranging from bioaccumulation of heavy metals to decreased ecological fitness. Conversely, AMLs were an overall benefit to imperiled bat populations and could serve as tools for conservation. Studies were unevenly distributed across different wildlife taxa groups, echoing the necessity for additional taxonomically diverse research. We suggest that standardized wildlife survey methods be used to assess how different species utilize AMLs. Federal and state agencies can use these surveys to establish effective remediation plans for individual AML sites and minimize the risks to both wildlife and humans.

Investigating the effects of three trace metals on the viability, embryonic development, and locomotor behavior of the Seminole ramshorn snail at environmentally relevant concentrations.

Trace metal contamination is a widespread issue due to its many natural and anthropogenic sources and known carcinogenic, teratogenic, and reproductive effects. As previous invertebrate trace metal research has primarily focused on model species (Daphnia magna, Chironomidae, etc.), our understanding of effects on non-model invertebrate species remains relatively poor. As such, this study assessed the exposure effects of cadmium, arsenic, and lead on viability, locomotor behavior, and embryonic development of the Seminole ramshorn snail (Planorbella duryi). Exposure treatments of CdCl2 , Na2 HAsO4 • 7H2 O, or Pb (NO3 )2 were prepared at concentrations of 0, 0.01, 0.1, 1, and 10 mg/L and confirmed using inductively coupled plasma optical emission spectroscopy (ICP-OES). Individual adult P. duryi were exposed for 7 days with viability assessed every 24 h, and locomotor behavior was accessed on Days 1 and 7 using ToxTrac v2.97 automated behavior software. Individual embryos from newly laid (<6 h old) embryonic clutches were exposed for 10 days, during which embryonic development stage was documented every 24 h. Based on our results, an additional follow-up study for cadmium was conducted using a lower range of 0-0.1 mg/L to allow for the observation of sublethal endpoints. Adult lead and cadmium exposure resulted in significant mortality in the highest treatments (1 and 10 mg/L), dose-dependent behavioral effects, and delayed embryonic development. Arsenic exposures resulted in little to no impacts for all assessed endpoints. Our results provide new insight into the sublethal impacts of these contaminants and highlight potential for behavior and embryonic development as useful tools for risk assessment. PRACTITIONER POINTS: The exposure effects of lead, cadmium, and arsenic on the viability, embryonic development, and locomotor behavior of a common freshwater snail species was investigated using environmentally relevant concentrations. The severity of impact differed for each trace metal, with cadmium being the most toxic and arsenic the least toxic at concentrations ranging from 0 to 10 mg/L. Embryonic development appeared to be the most sensitive endpoint of those tested in this study, suggesting that exposure may have prolonged effects that extend to population and community levels. The Seminole ramshorn snail serves as a sensitive alternative model species that can be used to assess the impacts of contaminants on freshwater invertebrates in future studies.

Toxicant Responses and Culturing Characteristics of Long-Term Laboratory-Reared and Field Populations of Ceriodaphnia dubia.

Ceriodaphnia dubia is a standardized test organism for regulatory toxicity testing of surface waters and commercial chemicals because of its simplicity to culture and responsiveness to toxicants. For testing convenience, C. dubia is often cultured for extended periods in the laboratory with little knowledge of the impact on subsequent generations. Extended laboratory rearing could impact how they respond to stressors and decrease the accuracy of test results. The present study investigated if C. dubia cultured for an extended period were representative of three recently collected field populations by comparing their culturing characteristics and sensitivities to toxicants. For culturing characteristics, the field cultures were more challenging because they had shorter body lengths, fewer neonates, and higher mortality rates than the laboratory culture. Comparative chronic toxicity tests with sodium chloride and the neonicotinoid insecticide thiamethoxam indicated that the laboratory and field organisms did not differ much in their toxicological responses but did differ in the variability of responses (percentage of coefficient of variation). The differences between the laboratory and field cultures found in the present study highlight the challenges of addressing discrepancies between laboratory and field applications in existing standardized methodologies. Environ Toxicol Chem 2024;43:159-169. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Contaminants of Concern and Spatiotemporal Metabolomic Changes in Quagga Mussels (Dreissena bugensis rostriformis) from the Milwaukee Estuary (Wisconsin, USA).

Environmental metabolomics has emerged as a promising technique in the field of biomonitoring and as an indicator of aquatic ecosystem health. In the Milwaukee Estuary (Wisconsin, USA), previous studies have used a nontargeted metabolomic approach to distinguish between zebra mussels (Dreissena polymorpha) collected from sites of varying contamination. To further elucidate the potential effects of contaminants on bivalve health in the Milwaukee Estuary, the present study adopted a caging approach to study the metabolome of quagga mussels (Dreissena bugensis rostriformis) deployed in six sites of varying contamination for 2, 5, or 55 days. Caged mussels were co-deployed with two types of passive sampler (polar organic chemical integrative samplers and semipermeable membrane devices) and data loggers. In conjunction, in situ quagga mussels were collected from the four sites studied previously and analyzed for residues of contaminants and metabolomics using a targeted approach. For the caging study, temporal differences in the metabolomic response were observed with few significant changes observed after 2 and 5 days, but larger differences (up to 97 significantly different metabolites) to the metabolome in all sites after 55 days. A suite of metabolic pathways were altered, including biosynthesis and metabolism of amino acids, and upmodulation of phospholipids at all sites, suggesting a potential biological influence such as gametogenesis. In the caging study, average temperatures appeared to have a greater effect on the metabolome than contaminants, despite a large concentration gradient in polycyclic aromatic hydrocarbons residues measured in passive samplers and mussel tissue. Conversely, significant differences between the metabolome of mussels collected in situ from all three contaminated sites and the offshore reference site were observed. Overall, these findings highlight the importance of contextualizing the effects of environmental conditions and reproductive processes on the metabolome of model organisms to facilitate the wider use of this technique for biomonitoring and environmental health assessments. Environ Toxicol Chem 2024;43:307-323. © 2023 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.

Beaver-related restoration: An opportunity for sandy lowland streams in a human-dominated landscape.

The expansion of beavers into human-dominated landscapes can help improve the ecological status of degraded streams. This study aimed to assess the changes in the ecological status of a degraded sandy-bottom stream under the influence of processes generated by the activity of a European beaver. We assumed that the processes in the beaver dam-and-pond complex significantly improved the physical, chemical, and bacteriological properties of water and improved the ecological status of a small lowland stream in a human-dominated landscape. To achieve these objectives, water for quality assessment, benthic invertebrates, and fish samples were collected, and the forms of land use and the natural values of the stream were analysed. In the upstream zone of the sandy-bottom stream flowing in a human-dominated landscape, the inflow of pollutants caused a significant reduction in water quality. The amount of nutrients was high and the amount of dissolved oxygen was low. Similarly, the number of meso- and psychrophilic bacteria was significantly higher than in the reference stream. The building activity of beavers initiated a series of positive environmental changes, and beaver ponds improved the self-purification of water in a polluted stream. In the lower part of the studied stream, after the stream passed through the beaver wetland system, the water quality improved, which was indicated by a significant improvement in physicochemical, microbiological, and biotic indicators, that is based on macrozoobenthos and fish. By constructing dams, beavers flooded large areas and created habitats for plant and animal species associated with wetlands, including amphibians, birds, and mammals. The improvement of the ecological status of the stream, combined with the preservation and restoration of wetland habitats in the catchment area, enables the use of ecosystem services provided by the streams in the human-dominated landscape and their surroundings.

Invertebrate dispersal by waterbird species in neotropical wetlands / A dispersão de invertebrados por aves aquáticas na região neotropical

Abstract Endozoochory by waterbirds is particularly relevant to the dispersal of non-flying aquatic invertebrates. This ecological function exercised by birds has been demonstrated in different biogeographical regions, but there are no studies for the neotropical region. In this work, we identified propagules of invertebrates in faeces of 14 syntopic South American waterbird species representing six families, and hatched additional invertebrates from cultured faeces. We tested whether propagule abundance, species richness and composition varied among bird species, and between the cold and warm seasons. We found 164 invertebrate propagules in faecal samples from seven different waterbirds species, including eggs of the Temnocephalida and Notonectidae, statoblasts of bryozoans (Plumatella sp.) and ephippia of Cladocera. Ciliates (including Paramecium sp. and Litostomatea), nematodes and rotifers (Adineta sp. and Nottomatidae) hatched from cultured samples. Potential for endozoochory was confirmed for 12 of 14 waterbird species. Our statistical models suggest that richness and abundance of propagules are associated with bird species and not affected by seasonality. Dispersal by endozoochory is potentially important to a broad variety of invertebrates, being promoted by waterbirds with different ecological and morphological traits, which are likely to drive the dispersal of invertebrates in neotropical wetlands.

Resumo A endozoocoria promovida por aves aquáticas é particularmente relevante para a dispersão de invertebrados aquáticos não-voadores. Essa função ecológica exercida pelas aves tem sido demonstrada para diferentes regiões biogeográficas, porém, não existem estudos para a região neotropical. Neste trabalho nós identificamos propágulos de invertebrados encontrados em fezes de 14 espécies sintópicas de aves aquáticas da América do Sul, representando seis famílias de aves, e também invertebrados emergidos de amostras fecais cultivadas em laboratório. Testamos se a abundância, riqueza de espécies e composição de propágulos de invertebrados variavam entre as espécies de aves e entre estações. Nós encontramos 164 propágulos de invertebrados em amostras fecais de sete espécies de aves, incluindo ovos de Temnocephalida e Notonectidae, estatoblastos de briozoários (Plumatella sp.) e efípios de Cladocera. Ciliados (incluindo Paramecium sp. e Litostomatea), nematóides e rotíferos (Adineta sp. e Nottomatidae) eclodiram de amostras cultivadas. O potencial para endozoocoria foi confirmado para 12 das 14 espécies de aves aquáticas investigadas. Nossos modelos estatísticos sugerem que a riqueza e abundância de propágulos estão associadas às espécies de aves e não são afetadas pela sazonalidade. A dispersão por endozoocoria é importante para uma ampla variedade de invertebrados, sendo promovida por aves aquáticas com diferentes características ecológicas e morfológicas as quais provavelmente regulam a dispersão de invertebrados entre áreas úmidas neotropicais.

Combined effects of climate warming and pharmaceuticals on a tri-trophic freshwater food web.

Multiple anthropogenic stressors influence the functioning of lakes and ponds, but their combined effects are often little understood. We conducted two mesocosm experiments to evaluate the effects of warming (+4 °C above ambient temperature) and environmentally relevant concentrations of a mixture of commonly used pharmaceuticals (cardiovascular, psychoactive, antihistamines, antibiotics) on tri-trophic food webs representative of communities in ponds and other small standing waters. Communities were constituted of phyto- and zooplankton and macroinvertebrates (molluscs and insects) including benthic detritivores, grazers, omnivorous scrapers, omnivorous piercers, water column predators, benthic predators, and phytophilous predators. We quantified the main and interactive effects of warming and pharmaceuticals on each trophic level in the pelagic community and attributed them to the direct effects of both stressors and the indirect effects arising through biotic interactions. Warming and pharmaceuticals had stronger effects in the summer experiment, altering zooplankton community composition and causing delayed or accelerated emergence of top insect predators (odonates). In the summer experiment, both stressors and top predators reduced the biomass of filter-feeding zooplankton (cladocerans), while warming and pharmaceuticals had opposite effects on phytoplankton. In the winter experiment, the effects were much weaker and were limited to a positive effect of warming on phytoplankton biomass. Overall, we show that pharmaceuticals can exacerbate the effects of climate warming in freshwater ecosystems, especially during the warm season. Our results demonstrate the utility of community-level studies across seasons for risk assessment of multiple emerging stressors in freshwater ecosystems.

Invertebrate dispersal by waterbird species in neotropical wetlands

Abstract Endozoochory by waterbirds is particularly relevant to the dispersal of non-flying aquatic invertebrates. This ecological function exercised by birds has been demonstrated in different biogeographical regions, but there are no studies for the neotropical region. In this work, we identified propagules of invertebrates in faeces of 14 syntopic South American waterbird species representing six families, and hatched additional invertebrates from cultured faeces. We tested whether propagule abundance, species richness and composition varied among bird species, and between the cold and warm seasons. We found 164 invertebrate propagules in faecal samples from seven different waterbirds species, including eggs of the Temnocephalida and Notonectidae, statoblasts of bryozoans (Plumatella sp.) and ephippia of Cladocera. Ciliates (including Paramecium sp. and Litostomatea), nematodes and rotifers (Adineta sp. and Nottomatidae) hatched from cultured samples. Potential for endozoochory was confirmed for 12 of 14 waterbird species. Our statistical models suggest that richness and abundance of propagules are associated with bird species and not affected by seasonality. Dispersal by endozoochory is potentially important to a broad variety of invertebrates, being promoted by waterbirds with different ecological and morphological traits, which are likely to drive the dispersal of invertebrates in neotropical wetlands.

Resumo A endozoocoria promovida por aves aquáticas é particularmente relevante para a dispersão de invertebrados aquáticos não-voadores. Essa função ecológica exercida pelas aves tem sido demonstrada para diferentes regiões biogeográficas, porém, não existem estudos para a região neotropical. Neste trabalho nós identificamos propágulos de invertebrados encontrados em fezes de 14 espécies sintópicas de aves aquáticas da América do Sul, representando seis famílias de aves, e também invertebrados emergidos de amostras fecais cultivadas em laboratório. Testamos se a abundância, riqueza de espécies e composição de propágulos de invertebrados variavam entre as espécies de aves e entre estações. Nós encontramos 164 propágulos de invertebrados em amostras fecais de sete espécies de aves, incluindo ovos de Temnocephalida e Notonectidae, estatoblastos de briozoários (Plumatella sp.) e efípios de Cladocera. Ciliados (incluindo Paramecium sp. e Litostomatea), nematóides e rotíferos (Adineta sp. e Nottomatidae) eclodiram de amostras cultivadas. O potencial para endozoocoria foi confirmado para 12 das 14 espécies de aves aquáticas investigadas. Nossos modelos estatísticos sugerem que a riqueza e abundância de propágulos estão associadas às espécies de aves e não são afetadas pela sazonalidade. A dispersão por endozoocoria é importante para uma ampla variedade de invertebrados, sendo promovida por aves aquáticas com diferentes características ecológicas e morfológicas as quais provavelmente regulam a dispersão de invertebrados entre áreas úmidas neotropicais.

When and How to Conduct Ecotoxicological Tests Using Natural Field-Collected Sediment.

In recent years, the sediment compartment has gained more attention when performing toxicity tests, with a growing emphasis on gaining more ecological relevance in testing. Though many standard guidelines recommend using artificially formulated sediment, most sediment studies are using natural sediment collected in the field. Although the use of natural field-collected sediment contributes to more environmentally realistic exposure scenarios and higher well-being for sediment-dwelling organisms, it lowers comparability and reproducibility among studies as a result of, for example, differences in the base sediment depending on sampling site, background contamination, particle size distribution, or organic matter content. The aim of this methodology contribution is to present and discuss best practices related to collecting, handling, describing, and applying natural field-collected sediment in ecotoxicological testing. We propose six recommendations: (1) natural sediment should be collected at a well-studied site, historically and by laboratory analysis; (2) larger quantities of sediment should be collected and stored prior to initiation of an experiment to ensure a uniform sediment base; (3) any sediment used in ecotoxicological testing should be characterized, at the very least, for its water content, organic matter content, pH, and particle size distribution; (4) select spiking method, equilibration time, and experimental setup based on the properties of the contaminant and the research question; (5) include control-, treated similarly to the spiked sediment, and solvent control sediment when appropriate; and (6) quantify experimental exposure concentrations in the overlying water, porewater (if applicable), and bulk sediment at least at the beginning and the end of each experiment. Environ Toxicol Chem 2023;00:1-10. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Optimizing Sex Ratios of Hyalella azteca to Reduce Variability in Reproduction and Improve Reproductive Toxicity Test Methods.

Standard toxicological test methods with Hyalella azteca incorporate both lethal and sublethal (growth, reproduction) endpoints, although lethal endpoints are often favored in routine testing. However, sublethal endpoints are important to consider because they are ecologically relevant and are often more sensitive than lethality. It is difficult to achieve robust data for reproduction in H. azteca because high biological variability is associated with reproductive yield, likely due to complex interactions in mate selection, which can add to the variability in brood sizes that females produce. In addition, effects on reproduction often co-occur with effects on growth, making it difficult to differentiate effects on growth from those on reproduction in standard tests initiated with juveniles. The present study characterized the reproductive capacity of H. azteca by investigating the role of sex ratios in reproductive yield. Experiments were initiated in the absence of toxicants with sexually mature (6-8-week-old) individuals that were placed in different female-to-male ratios (1:1, 2:3, 3:2, and 7:3). Reproduction was monitored during weekly static renewals for 7 weeks. The results indicated that a higher female:male ratio (seven females to three males) improved reproductive success and lowered biological variability in reproduction. In addition, the body lengths and weights of newly mature amphipods were used to create a size distribution to aid in the identification of small female amphipods. Based on the results of our study, a novel reproductive toxicity test method is proposed that begins with sexually mature amphipods in a ratio of seven females to three males to minimize reproductive variability in amphipods and to enable a more effective assessment of contaminants using reproduction as a highly sensitive endpoint. However, evaluation of this novel method with toxicants is needed to demonstrate that results are comparable to standard methods. Environ Toxicol Chem 2023;00:1-11. © 2023 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Daphnia magna Multigeneration Exposure to Carbendazim: Gene Transcription Responses.

The world population is experiencing colossal growth and thus demand for food, leading to an increase in the use of pesticides. Persistent pesticide contamination, such as carbendazim, remains a pressing environmental concern, with potentially long-term impacts on aquatic ecosystems. In the present study, Daphnia magna was exposed to carbendazim (5 µg L-1) for 12 generations, with the aim of assessing gene transcription alterations induced by carbendazim (using a D. magna custom microarray). The results showed that carbendazim caused changes in genes involved in the response to stress, DNA replication/repair, neurotransmission, ATP production, and lipid and carbohydrate metabolism at concentrations already found in the environment. These outcomes support the results of previous studies, in which carbendazim induced genotoxic effects and reproduction impairment (increasing the number of aborted eggs with the decreasing number of neonates produced). The exposure of daphnids to carbendazim did not cause a stable change in gene transcription between generations, with more genes being differentially expressed in the F0 generation than in the F12 generation. This could show some possible daphnid acclimation after 12 generations and is aligned with previous multigenerational studies where few ecotoxicological effects at the individual and populational levels and other subcellular level effects (e.g., biochemical biomarkers) were found.

Water beetles (Coleoptera) associated with Afrotemperate Forest patches in the Garden Route National Park, South Africa.

Southern Afrotemperate Forest is concentrated in the southern Cape region of South Africa and whilst it is relatively well known botanically, the fauna, specifically the aquatic invertebrate fauna, is poorly documented. The majority of remaining intact forest habitat is contained within the Garden Route National Park (GRNP), which straddles the provincial boundary between the Western and Eastern Cape. This study undertakes a survey of the water beetle fauna inhabiting the GRNP. The aquatic ecosystems within temperate forests of the region are poorly researched from an ecological and biodiversity perspective, despite being known to harbour endemic invertebrate elements. We collected water beetles and in situ physico-chemical data from a total of 31 waterbodies across the park over two seasons (summer and late winter) in 2017. The waterbodies sampled were mostly small freshwater perennial streams and isolated forest ponds. A total of 61 beetle taxa was recorded (29 Adephaga, 32 Polyphaga) from these waterbodies. The water beetle fauna of these forests appears to be diverse and contains many species endemic to the fynbos-dominated Cape Floristic Region, but very few of the species appear to be forest specialists. This is in contrast to the fynbos heathland habitat of the region, which harbours a high number of water beetle species endemic to this habitat, often with Gondwanan affinity. Our study is the first to document the water beetles of Afrotemperate Forests in the southern Cape region and provides an important baseline for future work on such habitats in the region and in other parts of southern Africa.

Methylation and Demethylation of Emerging Contaminants Changed Bioaccumulation and Acute Toxicity in Daphnia magna.

Contaminants of emerging concern (CECs) in the environment undergo various transformations, leading to the formation of transformation products (TPs) with a modified ecological risk potential. Although the environmental significance of TPs is increasingly recognized, there has been relatively little research to understand the influences of such transformations on subsequent ecotoxicological safety. In this study, we used four pairs of CECs and their methylated or demethylated derivatives as examples to characterize changes in bioaccumulation and acute toxicity in Daphnia magna, as a result of methylation or demethylation. The experimental results were further compared to quantitative structure-activity relationship (QSAR) predictions. The methylated counterpart in each pair generally showed greater acute toxicity in D. magna, which was attributed to their increased hydrophobicity. For example, the LC50 values of methylparaben (34.4 ± 4.3 mg L-1) and its demethylated product (225.6 ± 17.3 mg L-1) differed about eightfold in D. magna. The methylated derivative generally exhibited greater bioaccumulation than the demethylated counterpart. For instance, the bioaccumulation of methylated acetaminophen was about 33-fold greater than that of acetaminophen. In silico predictions via QSARs aligned well with the experimental results and suggested an increased persistence of the methylated forms. The study findings underline the consequences of simple changes in chemical structures induced by transformations such as methylation and demethylation and highlight the need to consider TPs to achieve a more holistic understanding of the environmental fate and risks of CECs.

Hypoxia in aquatic invertebrates: Occurrence and phenotypic and molecular responses.

Global deoxygenation in aquatic systems is an increasing environmental problem, and substantial oxygen loss has been reported. Aquatic animals have been continuously exposed to hypoxic environments, so-called "dead zones," in which severe die-offs among organisms are driven by low-oxygen events. Multiple studies of hypoxia exposure have focused on in vivo endpoints, metabolism, oxidative stress, and immune responses in aquatic invertebrates such as molluscs, crustaceans, echinoderms, and cnidarians. They have shown that acute and chronic exposure to hypoxia induces significant decreases in locomotion, respiration, feeding, growth, and reproduction rates. Also, several studies have examined the molecular responses of aquatic invertebrates, such as anaerobic metabolism, reactive oxygen species induction, increased antioxidant enzymes, immune response mechanisms, regulation of hypoxia-inducible factor 1-alpha (HIF-1α) genes, and differently expressed hemoglobin/hemocyanin. The genetic basis of those molecular responses involves HIF-1α pathway genes, which are highly expressed in hypoxic conditions. However, the identification of HIF-1α-related genes and understanding of their applications in some aquatic invertebrates remain inadequate. Also, some species of crustaceans, rotifers, sponges, and ctenophores that lack HIF-1α are thought to have alternative defense mechanisms to cope with hypoxia, but those factors are still unclear. This review covers the formation of hypoxia in aquatic environments and the various adverse effects of hypoxia on aquatic invertebrates. The limitations of current hypoxia research and genetic information about the HIF-1α pathway are also discussed. Finally, this paper explains the underlying processes of the hypoxia response and presents an integrated program for research about the molecular mechanisms of hypoxic stresses in aquatic invertebrates.

Respirometry reveals major lineage-based differences in the energetics of osmoregulation in aquatic invertebrates.

All freshwater organisms are challenged to control their internal balance of water and ions in strongly hypotonic environments. We compared the influence of external salinity on the oxygen consumption rates (MO2) of three species of freshwater insects, one snail and two crustaceans. Consistent with available literature, we found a clear decrease in MO2 with increasing salinity in the snail Elimia sp. and crustaceans Hyalella azteca and Gammarus pulex (r5=-0.90, P=0.03). However, we show here for the first time that metabolic rate was unchanged by salinity in the aquatic insects, whereas ion transport rates were positively correlated with higher salinities. In contrast, when we examined the ionic influx rates in the freshwater snail and crustaceans, we found that Ca uptake rates were highest under the most dilute conditions, while Na uptake rates increased with salinity. In G. pulex exposed to a serially diluted ion matrix, Ca uptake rates were positively associated with MO2 (r5=-0.93, P=0.02). This positive association between Ca uptake rate and MO2 was also observed when conductivity was held constant but Ca concentration was manipulated (1.7-17.3 mg Ca l-1) (r5=0.94, P=0.05). This finding potentially implicates the cost of calcium uptake as a driver of increased metabolic rate under dilute conditions in organisms with calcified exoskeletons and suggests major phyletic differences in osmoregulatory physiology. Freshwater insects may be energetically challenged by higher salinities, while lower salinities may be more challenging for other freshwater taxa.

Elucidating the spatial distribution of organic contaminants and their biotransformation products in amphipod tissue by MALDI- and DESI-MS-imaging.

The application of mass spectrometry imaging (MSI) is a promising tool to analyze the spatial distribution of organic contaminants in organisms and thereby improve the understanding of toxicokinetic and toxicodynamic processes. MSI is a common method in medical research but has been rarely applied in environmental science. In the present study, the suitability of MSI to assess the spatial distribution of organic contaminants and their biotransformation products (BTPs) in the aquatic invertebrate key species Gammarus pulex was studied. Gammarids were exposed to a mixture of common organic contaminants (carbamazepine, citalopram, cyprodinil, efavirenz, fluopyram and terbutryn). The distribution of the parent compounds and their BTPs in the organisms was analyzed by two MSI methods (MALDI- and DESI-HRMSI) after cryo-sectioning, and by LC-HRMS/MS after dissection into different organ compartments. The spatial distribution of contaminats in gammarid tissue could be successfully analyzed by the different analytical methods. The intestinal system was identified as the main site of biotransformation, possibly due to the presence of biotransforming enzymes. LC-HRMS/MS was more sensitive and provided higher confidence in BTP identification due to chromatographic separation and MS/MS. DESI was found to be the more sensitive MSI method for the analyzed contaminants, whereas additional biomarkers were found using MALDI. The results demonstrate the suitability of MSI for investigations on the spatial distribution of accumulated organic contaminants. However, both MSI methods required high exposure concentrations. Further improvements of ionization methods would be needed to address environmentally relevant concentrations.

Neonicotinoid mixture alters trophic interactions in a freshwater aquatic invertebrate community.

Neonicotinoids are increasingly and widely used systemic insecticides in agriculture, residential applications, and elsewhere. These pesticides can sometimes occur in small water bodies in exceptionally high concentrations, leading to downstream non-target aquatic toxicity. Although insects appear to be the most sensitive group to neonicotinoids, other aquatic invertebrates may also be affected. Most existing studies focus on single-insecticide exposure and very little is known concerning the impact of neonicotinoid mixtures on aquatic invertebrates at the community level. To address this data gap and explore community-level effects, we performed an outdoor mesocosm experiment that tested the effect of a mixture of three common neonicotinoids (formulated imidacloprid, clothianidin and thiamethoxam) on an aquatic invertebrate community. Exposure to the neonicotinoid mixture induced a top-down cascading effect on insect predators and zooplankton, ultimately increasing phytoplankton. Our results highlight complexities of mixture toxicity occurring in the environment that may be underestimated with traditional mono-specific toxicological approaches.

Laboratory-Derived Bioaccumulation Kinetic Parameters for Four Per- and Polyfluoroalkyl Substances in Freshwater Mussels.

Although freshwater mussels are imperiled and identified as key conservation priorities, limited bioaccumulation information is available on these organisms for contaminants of emerging concern. In the present study we investigated the bioaccumulation of per- and polyfluoroalkyl substances (PFAS) in the model freshwater pond mussel Sagittunio subrostratus because mussels provide important ecosystem services and are important components of aquatic systems where PFAS occur. In the present study we selected four representative perfluorinated carboxylic acids and sulfonic acids, then determined the bioaccumulation kinetics of freshwater mussels in a controlled laboratory study. Because uptake (ku ) and elimination (ke ) rate constants and time to steady state are important parameters for food web bioaccumulation models, we derived bioaccumulation kinetic parameters following exposure to perfluorohexane sulfonic acid (PFHxS), perfluorooctane sulfonic acid (PFOS), and perfluorodecanoic acid (PFDA) at 10 µg/L and perfluoroundecanoic acid (PFUnDA) at 1 µg/L during a 14-day uptake period followed by a 7-day elimination period. Kinetic and ratio-based bioaccumulation factors (BAFs) were subsequently calculated, for example ratio-based BAFs for mussel at day 7 were determined for PFHxS (0.24 ± 0.08 L/kg), PFOS (7.73 ± 1.23 L/kg), PFDA (4.80 ± 1.21 L/kg), and PFUnDA (84.0 ± 14.4 L/kg). We generally observed that, for these four model PFAS, freshwater mussels have relatively low BAF values compared with other aquatic invertebrates and fish. Environ Toxicol Chem 2023;42:1190-1198. © 2023 SETAC. This article has been contributed to by U.S. Government employees and their work is in the public domain in the USA.