Synaptic Mechanisms of Ethanol Tolerance and Neuroplasticity: Insights from Invertebrate Models.

Alcohol tolerance is a neuroadaptive response that leads to a reduction in the effects of alcohol caused by previous exposure. Tolerance plays a critical role in the development of alcohol use disorder (AUD) because it leads to the escalation of drinking and dependence. Understanding the molecular mechanisms underlying alcohol tolerance is therefore important for the development of effective therapeutics and for understanding addiction in general. This review explores the molecular basis of alcohol tolerance in invertebrate models, Drosophila and C. elegans, focusing on synaptic transmission. Both organisms exhibit biphasic responses to ethanol and develop tolerance similar to that of mammals. Furthermore, the availability of several genetic tools makes them a great candidate to study the molecular basis of ethanol response. Studies in invertebrate models show that tolerance involves conserved changes in the neurotransmitter systems, ion channels, and synaptic proteins. These neuroadaptive changes lead to a change in neuronal excitability, most likely to compensate for the enhanced inhibition by ethanol.

Field-realistic doses of the neonicotinoid acetamiprid impact natural soil arthropod community diversity and structure.

The neonicotinoid acetamiprid is used as a foliar insecticide spray, which results in direct exposure of a wide variety of soil organisms. Laboratory testing indicated that acetamiprid is toxic to the Collembola (springtails) species Folsomia candida, while Acari (mites) seem relatively insensitive to neonicotinoids. Since such opposing effects on different soil arthropods might imbalance natural arthropod communities, this study determined: (i) if field-realistic doses of acetamiprid affect the abundance and diversity in soil arthropod communities, and (ii) whether these potential effects are short-term or persist after degradation of acetamiprid. Intact soil cores collected from an untreated grassland field were placed in the mesocosm set up 'CLIMECS', and the naturally sourced communities were exposed to a control and increasing field-realistic doses of acetamiprid (i.e. 0, 0.05, 0.2, 0.8 mg a.s./kg dry soil). Before and 7 and 54 days after spraying the insecticide, the abundance of mites and springtails and springtail diversity were assessed. Springtail and mite abundances were similar at the start of the experiment, but springtail abundance was significantly lowered while mite abundance increased shortly after exposure to increasing doses of acetamiprid. At the highest dose, springtail numbers decreased by 53% on average while the number of mites increased by 26%. This effect was no longer visible after 54 days, suggesting recovery of the community as a whole reflected by observed changes in community dissimilarity: shortly after application springtail communities clearly diverged from the control in terms of species composition, while communities converged again in the long-term. With our results, we are the first to show that field-realistic applications of N-nitroguanidine neonicotinoids can significantly impact natural soil fauna communities, which might have implications for soil ecosystem functioning.

Thermal tolerance of giant salmonfly nymphs (Pteronarcys californica) varies across populations in a regulated river.

Warming of aquatic ecosystems is transforming the distribution, phenology and growth of the organisms dependent upon these ecosystems. Aquatic insects such as stoneflies are especially vulnerable to warming because the aquatic nymph stage of their life cycle depends on cool, well-oxygenated, flowing water habitat. We tracked thermal effects on available aerobic capacity of the aquatic nymph stage of an iconic and vulnerable stonefly species, the giant salmonfly (Pteronarcys californica), to compare habitat thermal regime measurements for two salmonfly populations from habitats separated by a gradient in summer weekly maximum temperatures. Contrary to expectations, the thermal optima range of the warmer habitat population was cooler than for the cooler habitat population. We posit that this unexpected interpopulation variation in thermal response is more strongly driven by diel and seasonal thermal variability than by the highest summer temperatures experienced within respective habitats. Additionally, we show that summer daily maximum temperatures could result in periodic limits in available aerobic capacity to support work of the warmer habitat nymphs and may be the mechanism underlying reduced abundance relative to the upstream cooler habitat population. Our findings provide insight into potential thermal and metabolic mechanisms that could regulate the success of ecological and culturally important aquatic insect species experiencing global change. We conclude that thermal regimes and thermal variation, not just mean and maximum temperatures, are critical drivers of aquatic insect responses to water temperatures.

Reproductive Capacity, but not Food Consumption, is Reduced by Continuous Exposure to Typical Genotoxic Stressor γ-Rays in the sentinel species Gammarus fossarum.

The long-term impacts of radiocontaminants (and the associated risks) for ecosystems are still subject to vast societal and scientific debate while wildlife is chronically exposed to various sources and levels of either environmental or anthropogenic ionizing radiation from the use of nuclear energy. The present study aimed to assess induced phenotypical responses in both male and female gammarids after short-term continuous γ-irradiation, acting as a typical well-characterized genotoxic stressor that can interact directly with living matter. In particular, we started characterizing the effects using standardized measurements for biological effects on few biological functions for this species, especially feeding inhibition tests, molting, and reproductive ability, which have already been proven for chemical substances and are likely to be disturbed by ionizing radiation. The results show no significant differences in terms of the survival of organisms (males and females), of their short-term food consumption which is linked to the general health status (males and females), and of the molting cycle (females). In contrast, exposure significantly affected fecundity (number of embryos produced) at the highest dose rates for irradiated females (51 mGy h-1) and males (5 and 51 mGy h-1). These results showed that, in gammarids, reproduction, which is a critical endpoint for population dynamics, is the most radiosensitive phenotypic endpoint, with significant effects recorded on male reproductive capacity, which is more sensitive than in females. Environ Toxicol Chem 2024;00:1-9. © 2024 SETAC.

The multifaceted effects of flavonoids on neuroplasticity.

 There has been a significant increase in the incidence of multiple neurodegenerative and terminal diseases in the human population with life expectancy increasing in the current times. This highlights the urgent need for a more comprehensive understanding of how different aspects of lifestyle, in particular diet, may affect neural functioning and consequently cognitive performance as well as in enhancing overall health. Flavonoids, found in a variety of fruits, vegetables, and derived beverages, provide a new avenue of research that shows a promising influence on different aspects of brain function. However, despite the promising evidence, most bioactive compounds lack strong clinical research efficacy. In the current scoping review, we highlight the effects of Flavonoids on cognition and neural plasticity across vertebrates and invertebrates with special emphasis on the studies conducted in the pond snail, Lymnaea stagnalis, which has emerged to be a functionally dynamic model for studies on learning and memory. In conclusion, we suggest future research directions and discuss the social, cultural, and ethnic dependencies of bioactive compounds that influence how these compounds are used and accepted globally. Bridging the gap between preclinical and clinical studies about the effects of bioactive natural compounds on brain health will surely lead to lifestyle choices such as dietary Flavonoids being used complementarily rather than as replacements to classical drugs bringing about a healthier future.

Combined toxicity of abamectin and carbendazim on enzymatic and transcriptional levels in the soil-earthworm microcosm.

To reveal the toxicological mechanisms of pesticide mixtures on soil organisms, this study concentrated on evaluating enzymatic activity and gene expression changes in the earthworm Eisenia fetida (Savigny 1826). Despite being frequently exposed to multiple pesticides, including the common combination of abamectin (ABA) and carbendazim (CAR), environmental organisms have primarily been studied for the effects of individual pesticides. Acute toxicity results exhibited that the combination of ABA and CAR caused a synergistic impact on E. fetida. The levels of MDA, ROS, T-SOD, and caspase3 demonstrated a significant increase across most individual and combined groups, indicating the induction of oxidative stress and cell death. Additionally, the expression of three genes (hsp70, gst, and crt) exhibited a significant decrease following exposure to individual pesticides and their combinations, pointing toward cellular damage and impaired detoxification function. In contrast, a noteworthy increase in ann expression was observed after exposure to both individual pesticides and their mixtures, suggesting the stimulation of reproductive capacity in E. fetida. The present findings contributed to a more comprehensive understanding of the potential toxicity mechanisms of the ABA and CAR mixture, specifically on oxidative stress, cell death, detoxification dysfunction, and reproductive capacity in earthworms. Collectively, these data offered valuable toxicological insights into the combined effects of pesticides on soil organisms, enhancing our understanding of the underlying risks associated with the coexistence of different pesticides in natural soil environments.

Characterization of Springtail (Arrhopalites caecus) for Use in Soil Ecotoxicity Testing.

Springtails (subclass: Collembola) represent one of the most extensively studied invertebrate groups in soil ecotoxicology. This is because of their ease of laboratory culture, significant ecological role, and sensitivity to environmental contaminants. Folsomia candida (family: Isotomidae) is a globally widespread parthenogenetic species that is prevalent in laboratory toxicity testing with springtails. Conversely, Arrhopalites caecus (family: Arrhopalitidae), a parthenogenic globular springtail species, remains untested in soil ecotoxicology. This species is found in diverse habitats, including cave systems and forest leaf litter, and has a global distribution. The sensitivity of A. caecus to environmental contaminants, such as neonicotinoid insecticides, as well as its life history and optimal culturing conditions, are largely unknown. The present study describes the establishment of a pure A. caecus laboratory culture and characterization of its life cycle and culturing conditions. We assessed the sensitivity of A. caecus to various insecticides, including exposures to the neonicotinoid thiamethoxam in soil and through a novel feeding assay as well as to clothianidin and cyantraniliprole in spiked soil exposures. In 7- and 14-day exposures to thiamethoxam in agricultural soil, the 50% lethal concentration (LC50) values were determined to be 0.129 mg/kg dry weight and 0.010 mg/kg dry weight, respectively. The 14-day LC50 for exposure to thiamethoxam via spiked food was determined to be 0.307 mg/kg dry weight. In addition, the 28-day 50% effect concentration for inhibition of juvenile production from cyantraniliprole exposure in the same soil type was 0.055 mg/kg dry weight. Challenges encountered in using this species included susceptibility to mite infestation and low adult survival rates in the 28-day cyantraniliprole test. Environ Toxicol Chem 2024;43:1820-1835. © 2024 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Guidance on assessing the potential impacts of selenium in freshwater ecosystems.

Despite decades of fate and effects studies, environmental selenium (Se) contamination and management remain an issue for many freshwater systems in North America. Several regulatory bodies have promulgated updated targets or management levels for Se; however, additional guidance on best practices for monitoring Se to protect freshwater aquatic life is warranted. In this article, we describe current approaches to assessing the ecological risks of Se in impaired freshwater systems and outline recommended methods for collecting and analyzing biological and abiotic samples and interpreting data. Because reproductive impairment of fish populations is most commonly used to determine the potential impacts of Se, several biological factors that could affect Se toxicity are explored, including diet, trophic positions, reproductive biology, body size and maturity, migratory movements, and use of seasonal habitats. Measuring Se concentrations in mature eggs is the most reliable metric for estimating potential reproductive impairment in fish populations because the range of toxicity thresholds is relatively narrow for all but a few tolerant fish species. In situations where collecting mature eggs is not feasible, we review the use of alternative fish tissue for estimating potential effects. Factors affecting Se uptake from freshwater are also considered with guidance on collecting abiotic (e.g., water and sediment) and biotic components of aquatic food webs (e.g., macroinvertebrates, biofilm). Integr Environ Assess Manag 2024;00:1-16. © 2024 SETAC.

Unveiling the underlying drivers of Phanerozoic marine diversification.

In investigating global patterns of biodiversity through deep time, many large-scale drivers of diversification have been proposed, both biotic and abiotic. However, few robust conclusions about these hypothesized effectors or their roles have been drawn. Here, we use a linear stochastic differential equation (SDE) framework to test for the presence of underlying drivers of diversification patterns before examining specific hypothesized drivers. Using a global dataset of observations of skeletonized marine fossils, we infer origination, extinction and sampling rates (collectively called fossil time series) throughout the Phanerozoic using a capture-mark-recapture approach. Using linear SDEs, we then compare models including and excluding hidden (i.e. unmeasured) drivers of these fossil time series. We find evidence of large-scale underlying drivers of marine Phanerozoic diversification rates and present quantitative characterizations of these. We then test whether changing global temperature, sea-level, marine sediment area or continental fragmentation could act as drivers of the fossil time series. We show that it is unlikely any of these four abiotic factors are the hidden drivers we identified, though there is evidence for correlative links between sediment area and origination/extinction rates. Our characterization of the hidden drivers of Phanerozoic diversification and sampling will aid in the search for their ultimate identities.

Immediate Impacts of Wildfires on Ground-dwelling macroinvertebrate Communities under Stones in Mediterranean Oak Forests.

Wildfires are considered a major disturbance to forest ecosystems in the Mediterranean countries of Southern Europe. Although ground-dwelling macroinvertebrates are crucial to many soil functions, there is a fundamental lack of understanding of how wildfires impact this community in the immediate term and of the role of stones in their survival. Hence, in the present study we assessed the immediate effects of wildfires in the ground-dwelling macroinvertebrate community found under stones by comparing communities in burnt and non-burnt Mediterranean oak forests. Our results revealed that stones allowed the survival of many taxa in the burnt area. However, abundance, richness, diversity, and equitability per stone were significantly lower at the burnt than unburnt sites. Furthermore, the results also showed that richness and abundance increased significantly with increasing stone depth and area, both at the burnt and unburnt sites. Significant changes at the trophic level were observed in the burnt area comparing to the unburnt, particularly a decline in predators. No significant differences were identified concerning habitat associations among taxa. Overall, this study stressed the role of stones as microhabitats and refuge for the ground-dwelling macroinvertebrate community during wildfires.

Exploring BPA alternatives - Environmental levels and toxicity review.

Bisphenol A alternatives are manufactured as potentially less harmful substitutes of bisphenol A (BPA) that offer similar functionality. These alternatives are already in the market, entering the environment and thus raising ecological concerns. However, it can be expected that levels of BPA alternatives will dominate in the future, they are limited information on their environmental safety. The EU PARC project highlights BPA alternatives as priority chemicals and consolidates information on BPA alternatives, with a focus on environmental relevance and on the identification of the research gaps. The review highlighted aspects and future perspectives. In brief, an extension of environmental monitoring is crucial, extending it to cover BPA alternatives to track their levels and facilitate the timely implementation of mitigation measures. The biological activity has been studied for BPA alternatives, but in a non-systematic way and prioritized a limited number of chemicals. For several BPA alternatives, the data has already provided substantial evidence regarding their potential harm to the environment. We stress the importance of conducting more comprehensive assessments that go beyond the traditional reproductive studies and focus on overlooked relevant endpoints. Future research should also consider mixture effects, realistic environmental concentrations, and the long-term consequences on biota and ecosystems.

Progress in the environmental risk assessment of plant protection products in Brazil: An overview of aquatic organism proposals.

Since 2019, the Brazilian Institute of Environment and Renewable Natural Resources (IBAMA) has actively developed pesticide environmental risk assessment (ERA) frameworks adapted to Brazil's specific ecological contexts. This endeavor, supported by funding from the Brazilian Ministry of Justice and in partnership with academic institutions, has led to a concerted effort to establish ERA protocols for various taxa, including birds and mammals, soil organisms, aquatic organisms, and reptiles and amphibians. The outcomes of this initiative were conveyed in two workshops held in February and November 2023, during which the agency communicated its findings to the technical-regulatory community. This article represents one of three articles that provide more detailed insights into the ERA propositions for all taxa. In this article, we summarize the proposals for aquatic organisms presented and discussed during the workshops, which can be used as an informational source by the technical-regulatory community. Integr Environ Assess Manag 2024;00:1-6. © 2024 The Author(s). Integrated Environmental Assessment and Management published by Wiley Periodicals LLC on behalf of Society of Environmental Toxicology & Chemistry (SETAC).

Inferential reasoning abilities in wild-caught bumblebees.

The ability to make a decision by excluding alternatives (i.e. inferential reasoning) is a type of logical reasoning that allows organisms to solve problems with incomplete information. Several species of vertebrates have been shown to find hidden food using inferential reasoning abilities. Yet little is known about invertebrates' logical reasoning capabilities. In three experiments, I examined wild-caught bumblebees' abilities to locate a 'rewarded' stimulus using direct information or incomplete information-the latter requiring bees to use inferential reasoning. To do so, I adapted three paradigms previously used with primates-the two-cup, three-cup and double two-cup tasks. Bumblebees saw either two paper strips (experiment 1), three paper strips (experiment 2) or two pairs of paper strips (experiment 3) and experienced one of them being rewarded or unrewarded. At the test, they could choose between two (experiment 1), three (experiment 2) or four paper strips (experiment 3). Bumblebees succeeded in the three tasks and their performance was consistent with inferential reasoning. These findings highlight the importance of comparative studies with invertebrates to comprehensively track the evolution of reasoning abilities, in particular, and cognition, in general.

Impact of pesticides on non-target invertebrates in agricultural ecosystems.

In fact, less than 1% of applied pesticides reach their target pests, while the remainder pollute the neighboring environment and adversely impact human health as well as non-target organisms in agricultural ecosystem. Pesticides can contribute to the loss of agrobiodiversity, which are essential to maintaining the agro-ecosystem's structure and functioning in order to produce and secure enough food. This review article examines the negative effects of pesticides on non-target invertebrates including earthworms, honeybees, predators, and parasitoids. It also highlights areas where further research is needed to address unresolved issues related to pesticide exposure, aiming to improve conservation efforts for these crucial species. These organisms play crucial roles in ecosystem functioning, such as soil health, pollination, and pest control. Both lethal and sub-lethal effects of pesticides on the selected non-target invertebrates were discussed. Pesticides affect DNA integrity, enzyme activity, growth, behavior, and reproduction of earthworms even at low concentrations. Pesticides could also induce a reduction in individual survival, disruption in learning performance and memory, as well as a change in the foraging behavior of honeybees. Additionally, pesticides adversely affect population growth indices, reproduction, development, longevity, and consumption of predators and parasitoids. As a result, pesticides must pass adequate ecotoxicological risk assessment to be enlisted by regulatory authorities. Therefore, it is important to adopt integrated pest management (IPM) strategies that minimize pesticide use and promote the conservation of beneficial organisms in order to maintain agrobiodiversity and sustainable agricultural systems. Furthermore, adopting precision agriculture and organic farming lessen these negative effects as well.less than.

Noise accelerates embryonic development in a key crab species: Morphological and physiological carryover effects on early life stages.

Anthropogenic noise is considered one important global pollutant. The impact of noise on marine invertebrates has been less assessed. The present study evaluated the chronic effect of the motorboat noise obtained from a lagoon's soundscape, the natural habitat of the key crab Neohelice granulata, on its whole embryonic development, considering morphological and physiological carryover effects on embryos and hatched larvae. Results demonstrated that embryonic development was shortened under noise exposure. The effects on advanced embryos, larvae and adult females were: increased heartbeats and non-viable eggs, and decreased fecundity. Biochemical responses showed lipid peroxidation in embryos while antioxidant enzymes were activated in larvae and adults, indicating a counteracting effect related to the life stage. The negative effects on fitness offspring may imply ecological consequences at the population level. Results are discussed in terms of the ecosystem engineer species studied and the habitat, a MAB UNESCO Reserve lagoon, suggesting the urgent need to develop mitigation plans.

See-Star: a versatile hydrogel-based protocol for clearing large, opaque and calcified marine invertebrates.

Studies of morphology and developmental patterning in adult stages of many invertebrates are hindered by opaque structures, such as shells, skeletal elements, and pigment granules that block or refract light and necessitate sectioning for observation of internal features. An inherent challenge in studies relying on surgical approaches is that cutting tissue is semi-destructive, and delicate structures, such as axonal processes within neural networks, are computationally challenging to reconstruct once disrupted. To address this problem, we developed See-Star, a hydrogel-based tissue clearing protocol to render the bodies of opaque and calcified invertebrates optically transparent while preserving their anatomy in an unperturbed state, facilitating molecular labeling and observation of intact organ systems. The resulting protocol can clear large (> 1 cm3) specimens to enable deep-tissue imaging, and is compatible with molecular techniques, such as immunohistochemistry and in situ hybridization to visualize protein and mRNA localization. To test the utility of this method, we performed a whole-mount imaging study of intact nervous systems in juvenile echinoderms and molluscs and demonstrate that See-Star allows for comparative studies to be extended far into development, facilitating insights into the anatomy of juveniles and adults that are usually not amenable to whole-mount imaging.

Baseline Inventory of Benthic Macrofauna in German Marine Protected Areas (2020-2022) before Closure for Bottom-Contact Fishing.

The response of benthic habitats and organisms to bottom-contact fishing intensity is investigated in marine protected areas (MPAs) of the German EEZ in the North and Baltic Seas. We examined the current state of macrofauna biodiversity in 2020-2022. Comparative analysis for macrofauna (in- and epifauna) inhabiting nine Natura 2000 MPAs constitutes a baseline to assess the effects of bottom-contact fishing exclusion in the future. Aspects of spatial and temporal variability are briefly summarized and discussed. We provide a species list for each region, including 481 taxa, of which 79 were found in both regions, 183 only in the North Sea, and 219 only in the Baltic Sea. The Baltic Sea dataset surprisingly included higher numbers of taxa and revealed more Red List species. The share of major taxonomic groups (polychaetes, bivalves and amphipods) in species richness showed peculiar commonalities between the two regions. In the North Sea, multivariate analysis of community structure revealed significantly higher within-similarity and stronger separation between the considered MPAs compared to the Baltic MPAs. Salinity, temperature and sediment fractions of sand were responsible for over 60% of the variation in the North Sea macrofauna occurrence data. Salinity, mud fraction and bottom-contact fishing were the most important drivers in the Baltic Sea and, together with other considered environmental drivers, were responsible for 53% of the variation. This study identifies aspects of macrofauna occurrence that may be used to assess (causes of) future changes.

An evaluation of fish and invertebrate mercury concentrations in the Caribbean Region.

Mercury is a ubiquitous pollutant of global concern but the threat of exposure is not homogenously distributed at local, regional, or global scales. The primary route of human exposure to mercury is through consumption of aquatic foods, which are culturally and economically important in the wider Caribbean Region, especially for Small Island Developing States (SIDS). We compiled more than 1600 samples of 108 unique species of fish and aquatic invertebrates collected between 2005 and 2023 from eleven countries or territories in the wider Caribbean Region. There was wide variability in total mercury concentrations with 55% of samples below the 0.23 µg/g wet weight (ww) guideline from the U.S. FDA/EPA (2022) for 2 or 3 weekly servings and 26% exceeding the 0.46 µg/g ww guideline consistent with adverse effects on human health from continual consumption, particularly for sensitive populations. Significant relationships were found between total mercury concentrations and taxonomic family, sampling country, fish length, and trophic level. The data analyzed here support the need for further sampling with concrete geospatial data to better understand patterns and mechanisms in mercury concentrations and allow for more informed decision making on the consumption of fish and invertebrates from the wider Caribbean Region as well as supporting efforts to evaluate the effectiveness of national, regional, and international mercury policies.

Effects of the insecticide imidacloprid on aquatic invertebrate communities of the Ecuadorian Amazon.

Imidacloprid is a neonicotinoid insecticide that has received particular attention due to its widespread use and potential adverse effects for aquatic and terrestrial ecosystems. Its toxicity to aquatic organisms has been evaluated in central and southern Europe as well as in (sub-)tropical regions of Africa and Asia, showing high toxic potential for some aquatic insects and zooplankton taxa. However, its toxicity to aquatic organisms representative of tropical regions of Latin America has never been evaluated. To fill this knowledge gap, we carried out a mesocosm experiment to assess the short- and long-term effects of imidacloprid on freshwater invertebrate communities representative of the Ecuadorian Amazon. A mesocosm experiment was conducted with five weekly applications of imidacloprid at four nominal concentrations (0.01 µg/L, 0.1 µg/L, 1 µg/L and 10 µg/L). Toxic effects were evaluated on zooplankton and macroinvertebrate populations and communities, as well as on water quality parameters for 70 days. Given the climatic conditions prevailing in the study area, characterized by a high solar radiation and abundant rainfall that resulted in mesocosm overflow, there was a rapid dissipation of the test compound from the water column (half-life: 4 days). The macroinvertebrate taxa Callibaetis pictus (Ephemeroptera), Chironomus sp. (Diptera), and the zooplankton taxon Macrocyclops sp., showed population declines caused by the imidacloprid treatment, with a 21-d Time Weighted Average No Observed Effect Concentrations (21-d TWA NOEC) of 0.46 µg/L, except for C. pictus which presented a 21-d TWA NOEC of 0.05 µg/L. In general terms, the sensitivity of these taxa to imidacloprid was greater than that reported for surrogate taxa in temperate zones and similar to that reported in other (sub-)tropical regions. These results confirm the high sensitivity of tropical aquatic invertebrates to this compound and suggest the need to establish regulations for the control of imidacloprid contamination in Amazonian freshwater ecosystems.

Environmentally relevant concentrations of the tricyclic antidepressant, amitriptyline, affect feeding and reproduction in a freshwater mollusc.

Antidepressant drugs (ADDs) are one of the most extensively used pharmaceuticals globally. They act at particularly low therapeutic concentrations to modulate monoamine neurotransmission, which is one of the most evolutionary conserved pathways in both humans and animal species including invertebrates. As ADDs are widely detected in the aquatic environment at low concentrations (ng/L to low µg/L), their potential to exert drug-target mediated effects in aquatic species has raised serious concerns. Amitriptyline (AMI) is the most widely used tricyclic ADD, while monoamines, the target of ADDs, are major bioregulators of multiple key physiological processes including feeding, reproduction and behaviour in molluscs. However, the effects of AMI on feeding, reproduction and mating behaviour are unknown in molluscs despite their ecological importance, diversity and reported sensitivity to ADDs. To address this knowledge gap, we investigated the effects of environmentally relevant concentrations of AMI (0, 10, 100, 500 and 1000 ng/L) on feeding, reproduction and key locomotor behaviours, including mating, in the freshwater gastropod, Biomphalaria glabrata over a period of 28 days. To further provide insight into the sensitivity of molluscs to ADDs, AMI concentrations (exposure water and hemolymph) were determined using a novel extraction method. The Fish Plasma Model (FPM), a critical tool for prioritization assessment of pharmaceuticals with potential to cause drug target-mediated effects in fish, was then evaluated for its applicability to molluscs for the first time. Disruption of food intake (1000 ng/L) and reproductive output (500 and 1000 ng/L) were observed at particularly low hemolymph levels of AMI, whereas locomotor behaviours were unaffected. Importantly, the predicted hemolymph levels of AMI using the FPM agreed closely with the measured levels. The findings suggest that hemolymph levels of AMI may be a useful indicator of feeding and reproductive disruptions in wild population of freshwater gastropods, and confirm the applicability of the FPM to molluscs for comparative pharmaceutical hazard identification.