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.

Gonad pathology, sex hormone modulation and vitellogenin expression in Chrysichthys nigrodigitatus from Lagos and Epe lagoons within the southern-lagoon system, Nigeria.

Introduction: Estrogenic chemicals in aquatic environments impact fish reproductive health, with vitellogenin protein levels serving as a crucial biomarker for xenoestrogen exposure. Limited knowledge exists on estrogenic effects in tropical environments, prompting an investigation into the influence of environmental estrogens on Chrysichthys nigrodigitatus in Lagos and Epe lagoons. Methods: A total of 195 fish samples underwent analysis for vitellogenin protein, sex hormones (testosterone and 17 ß-estradiol), and gonad pathology in effluent-receiving areas of the specified lagoons. Results: Gonadal alterations were observed in male and female fish, including empty seminiferous tubules and distorted ovaries. Intersex occurred in 3.81% of Lagos and 3.33% of Epe. Testosterone levels were generally higher in females and males from both lagoons, while E2 levels were higher in females from both lagoons, with Lagos showing higher levels than Epe. Vtg levels were higher in males than females in Lagos samples but showed no significant difference in Epe samples. Discussion: Contaminant analysis revealed similar trends in metals (Hg, As, Cr) and phthalates (DEHP, DBP, DEP) in both sexes in the Epe population. Multivariate depictions from the PCA showed sex-specific patterns of metal uptake (Cd) in male fishes at the Lagos Lagoon. The positive association between higher pH loadings and metal and DBP levels in sediment at the Lagos lagoon suggests the influence of higher alkalinity in lower bioavailability of contaminants. Conclusion: Endocrine disrupting effects were observed in male and female Chrysichthys nigrodigitatus in Lagos and Epe lagoons populations, with notable differences in hormone and contaminant concentrations between the two lagoon systems. Identification of specific contaminants and their spatial and temporal trends can inform targeted management and remediation efforts to protect and restore these valuable aquatic ecosystems.

Understanding target-specific effects of antidepressant drug pollution on molluscs: A systematic review protocol.

BACKGROUND: The environmental prevalence of widely prescribed human pharmaceuticals that target key evolutionary conserved biomolecules present across phyla is concerning. Antidepressants, one of the most widely consumed pharmaceuticals globally, have been developed to target biomolecules modulating monoaminergic neurotransmission, thus interfering with the endogenous regulation of multiple key neurophysiological processes. Furthermore, rising prescription and consumption rates of antidepressants caused by the burgeoning incidence of depression is consistent with increasing reports of antidepressant detection in aquatic environments worldwide. Consequently, there are growing concerns that long-term exposure to environmental levels of antidepressants may cause adverse drug target-specific effects on non-target aquatic organisms. While these concerns have resulted in a considerable body of research addressing a range of toxicological endpoints, drug target-specific effects of environmental levels of different classes of antidepressants in non-target aquatic organisms remain to be understood. Interestingly, evidence suggests that molluscs may be more vulnerable to the effects of antidepressants than any other animal phylum, making them invaluable in understanding the effects of antidepressants on wildlife. Here, a protocol for the systematic review of literature to understand drug target-specific effects of environmental levels of different classes of antidepressants on aquatic molluscs is described. The study will provide critical insight needed to understand and characterize effects of antidepressants relevant to regulatory risk assessment decision-making, and/or direct future research efforts. METHODS: The systematic review will be conducted in line with the guidelines by the Collaboration for Environmental Evidence (CEE). A literature search on Scopus, Web of Science, PubMed, as well as grey literature databases, will be carried out. Using predefined criteria, study selection, critical appraisal and data extraction will be done by multiple reviewers with a web-based evidence synthesis platform. A narrative synthesis of outcomes of selected studies will be presented. The protocol has been registered in the Open Science Framework (OSF) registry with the registration DOI: 10.17605/OSF.IO/P4H8W.

Endocrine disruptor responses in African sharptooth catfish (Clarias gariepinus) exposed to di-(2-ethylhexyl)-phthalate.

In the present study, we have investigated the endocrine disruptive effects of waterborne di-(2-ethylhexyl) phthalate (DEHP: 0 (control), 10, 100, 200 and 400 µg/L) on juvenile Clarias gariepinus by analyzing transcript patterns for hepatic vitellogenin (vtg), estrogen receptor-α (er-α), aromatase (cyp19a1b) and peroxisome proliferator activated receptor-α (ppar-α) using quantitative real-time PCR after 3, 7 and 14 days exposure period. In addition, we analyzed CYP19 and PPAR protein levels using enzyme-linked immunosorbent assay (ELISA), while cellular testosterone (T) and 17ß-estradiol (E2) levels were measured using enzyme immune assay (EIA). Tissue burden of DEHP was measured in the liver using gas chromatography-mass spectroscopy (GC-MS). We observed apparent concentration- and time-dependent increases of vtg, er-α, cyp19a1b transcripts, E2 and T levels after exposure to DEHP. A biphasic pattern of effect was observed for ppar-α, showing a concentration-dependent increase that peaked at 100 µg/L and thereafter, an apparent concentration-dependent decrease at 200 and 400 µg/L at all exposure days. Given that the post-differentiation changes of gonads in C. gariepinus corresponded with the 14-day sampling period, we separated the analyzed data into sexes after histological examination of the gonads, showing that females responded stronger, compared to males, to DEHP exposure at all exposure concentrations. Oocyte atresia, intersex (ovotestis) and karyoplasmic clumping were observed in females while male fish showed distortion and degeneration of seminiferous tubules and condensation of tubular cells in the 400 µg/L exposure group after 14 days. Corresponding canonical analysis (CCA) of all analyzed variables revealed a positive relationship between analyzed biological variables with increasing DEHP concentrations. Overall, molecular, biochemical and physiological responses presented in the present study indicate that exposure of C. gariepinus to waterborne DEHP produced endocrine disruptive responses with potential consequences for overt reproduction, development, physiology and general health of fish populations inhabiting phthalate contaminated aquatic environments. These responses represent valuable and effective biomarkers of exposure and effects, that can be adopted for screening the presence of EDCs in Nigeria and other developing countries.