Silver barb Barbodes gonionotus fry were exposed to polyvinyl chloride (PVC) fragments at increasing concentrations of 0.2, 0.5 and 1.0 mg/L for 96 h, following which whole body histological evaluation and analysis of the digestive enzymes trypsin and chymotrypsin were performed. Whole body trypsin and chymotrypsin activities increased significantly in fish exposed to 0.5 and 1.0 mg/L PVC as compared those exposed to zero or 0.2 mg/L PVC. In fish exposed to all tested concentrations, PVCs were observed in both the proximal and distal intestine, and fish exposed to 0.5–1.0 and 1.0 mg/L PVC, respectively, and these particles were associated with localized thickening of the mucosal epithelium. No tissue damage was evident in any other internal organs or gills. This lack of damage may be attributed to the absence of contaminants associated with the PVC fragments and their relatively smooth surface. The increased whole body trypsin and chymotrypsin activities may indicate an attempt to enhance digestion to compensate for epithelial thickening of the intestine and/or to digest the plastics.

Integrated compensatory responses of physiological systems towards homeostasis are generally overlooked when it comes to analysing alterations in biochemical parameters indicative of such processes. Here an hypothesis-driven multivariate analysis accounting for interactive multibiomarker responses was used to investigate effects of long-term exposure of Carcinus maenas to Hazardous and Noxious Substances (HNS). Adult male crabs were exposed to low and high post-spill levels of acrylonitrile (ACN) or aniline (ANL) for 21d. Bioaccumulation, feeding behaviour, and biomarkers related to mode-of-action (MoA) (detoxification, neurotransmission and energy production) were evaluated over time. Distinct temporal patterns of response to low and high exposure concentrations were depicted, with a main set of interactive multibiomarker predictors identified for each HNS (five for ACN and three for ANL), useful to follow coupled evolvement of biomarker responses. ACN caused peripheral neurotoxic effects coupled with enhanced biotransformation and significant oxidative damage particularly relevant in gills. ANL elicited alterations in central neurotransmission affecting ventilation coupled with very low levels of oxidative damage in gills. Results indicate chronic toxicity data are determinant to improve HNS hazard assessment if the aim is to obtain reliable risk calculations, and develop effective predictive models avoiding overestimation but sufficiently protective. Accounting for multibiomarker interactions brought otherwise overlooked information about C. maenas responses and MoA of ACN and ANL.

Since only a few studies have investigated effects of microplastics (MPs) by routes other than ingestion, this study was designed to analyze the accumulation patterns and transfer of toxic substances associated with microplastic exposure by simple attachment to (1) adult zebrafish (Danio rerio) gills and (2) zebrafish embryos. Two sizes of fluorescently labelled polymers (1–5 and 10–20 μm) loaded with the model polycyclic aromatic hydrocarbon (PAH) benzo[a]pyrene (BaP) were used to analyze fate, accumulation and transfer of microplastic-associated persistent organic pollutants (POPs) on gills and embryos.Results indicate that microplastics did not permanently accumulate at high amounts in adult zebrafish gills after 6 nor 24 h of incubation: Most particles only superficially adhered to the mucus layer on the filaments, which is constantly being excreted. In contrast, the smaller and heavier MPs (1–5 μm) accumulated in high numbers on the surface of zebrafish egg chorions. In both exposure scenarios, transfer of BaP could be visualized with fluorescence microscopy: A prominent BaP signal was visible both in gill filaments and arches after 6 and 24 h incubation and in zebrafish embryos after exposure to BaP-spiked microplastics. Furthermore, the gill EROD (Ethoxyresorufin-O-deethylase) assay showed a clear trend to CYP 1A (Cytochrom P450 1 A) induction via exposure to BaP-spiked microplastics. However, BaP from spiked microplastics did not reach sufficiently high concentrations to be able to induce morphological effects in the fish embryo toxicity test (FET). In contrast, control exposure to waterborne BaP did induce effects in the FET.As a conclusion, microplastics can also transfer POPs not only via ingestion, but also by simple attachment to epithelia or via the water column. However, further studies are needed to clarify if these interactions are of environmental concern relative to waterborne exposure to toxic substances.

Glyphosate (GLY) is one of the most used herbicide worldwide. Considering that information concerning the impact of GLY on bivalves is scarce, in this study we evaluated for the first time the effects of environmentally realistic concentrations of GLY (10, 100 and 1000 μg/L) to the mussel Mytilus galloprovincialis. Mussels were exposed for 7, 14 and 21 days and several biomarkers were measured in haemocytes/haemolymph (total haemocyte counts, haemocyte diameter and volume, haemolymph pH, haemolymph lactate dehydrogenase activity, haemocyte lysate lysozyme and acid phosphatase activities), as well as in gills and digestive gland (antioxidant enzyme and acetylcholinesterase activities). The concentrations of GLY and its main metabolite aminomethylphosphonic acid in the experimental tanks were also measured. The MANOVA analysis demonstrated that the experimental variables considered (exposure concentration, exposure duration, and their interaction) affected significantly biomarker responses. In addition, the two-way ANOVA analysis indicated that GLY was able to affect most of the cellular parameters measured, whereas antioxidant enzyme activities resulted to be influenced moderately. Interestingly, exposure to GLY reduced significantly acetylcholinesterase activity in gills. Although preliminary, the results of this study demonstrated that GLY can affect both cellular and biochemical parameters in mussels, highlighting a potential risk for aquatic invertebrates.

CO2-driven acidification and emerging contaminants, such as pharmaceuticals, pose new threats for the maintenance of natural populations of marine organisms by interfering with their normal biochemical pathways and defences. The combined effects of seawater acidification, as predicted in climate change scenarios, and an emerging contaminant (the non-steroidal anti-inflammatory drug, NSAID, diclofenac) on oxidative stress-related parameters were investigated in the Mediterranean mussel Mytilus galloprovincialis and the Manila clam Ruditapes philippinarum. A flow-through system was used to carry out a three-week exposure experiment with the bivalves. First, the animals were exposed to only three pH values for 7 days. The pH was manipulated by dissolving CO2 in the seawater to obtain two reduced pH treatments (pH −0.4 units and pH −0.7 units), which were compared with seawater at the natural pH level (8.1). Thereafter, the bivalves were concomitantly exposed to the three experimental pH values and environmentally relevant concentrations of diclofenac (0.00, 0.05 and 0.50 μg/L) for an additional 14 days. The activities of superoxide dismutase, catalase and cyclooxygenase, and lipid peroxidation and DNA strand-break formation were measured in both the gills and digestive gland after 7, 14 and 21 days of exposure to each experimental condition. The results show that the biochemical parameters measured in both the mussels and clams were more influenced by the reduced pH than by the contaminant or the pH*contaminant interaction, although the biomarker variation patterns differed depending on the species and tissues analysed. Generally, due to increases in its antioxidant defence, M. galloprovincialis was more resistant than R. philippinarum to both diclofenac exposure and reduced pH. Conversely, reduced pH induced a significant decrease in COX activity in both the gills and digestive gland of clams, possibly resulting in the increased DNA damage observed in the digestive gland tissue.

The goal of this study was to evaluate the adverse effects of wastewater effluents on freshwater crucian carp, Carassius auratus, inhabiting Sincheon stream using the integrated biomarker response (IBR) at the genotoxic (micronucleus [MN] test), oxidative stress (activity of catalase [CAT] and glutathione S-transferase [GST], and level of lipid peroxidation [LPO]), histopathological (degree of tissue changes [DTC]), and physiological (condition factor [CF] and liver somatic index [LSI]) levels. The CF and LSI were significantly (p < 0.05) enhanced in fish from downstream sites (DS1 and DS2) as compared to that of upstream (US) fish samples. Moreover, a significant increase in morphometric indices (DTC) was observed in C. auratus collected from downstream sites (p < 0.05) and histopathological responses showed the degree of pathogenicity in the order of liver > kidney > gills. The activities of CAT, GST, and LPO in fish from the DS1 and DS2 sites were notably increased in gills, liver, and kidney compared to that of fish from the US site. Additionally, the MN test level in C. auratus from the DS1 and DS2 were significantly increased (p < 0.05) when compared with that of the US site. Considering the higher bioaccumulation of Cd, Co, Cr, Mn, Ni, and Pb in gills, liver, and kidney of C. auratus collected from downstream sites compared to that of the upstream site (p < 0.05), the observed toxicity was likely attributable to metal accumulation. The multi-level IBR index was higher at the DS1 site (15.08) than at the DS2 (1.02) and the reference US (0.00) sites. Therefore, these findings demonstrated that wastewater effluent discharge induces significant DNA damage, oxidative stress, and tissue injuries in C. auratus and suggested that the multi-level IBR approach should be used to quantify these effects on fish in streams and rivers.

While the presence of microplastics (MPs) in marine environments has been detected worldwide, the importance of MPs pollution in freshwater environments has also been emphasized in recent years. However, the body of knowledge regarding the biological effects of MPs on freshwater organisms is still much more limited than on marine organisms. The aim of the present study was to evaluate the accumulation and tissue distribution of MPs in the freshwater fish red tilapia (Oreochromis niloticus), as well as the biochemical effects of MPs on O. niloticus. During 14 days of exposure to 0.1 μm polystyrene-MPs at concentrations of 1, 10, and 100 μg L−1, the MPs concentrations in various tissues of O. niloticus generally increased over time following the order gut > gills > liver ≈ brain. Moreover, the acetylcholinesterase (AChE) activity in the fish brain was inhibited by MPs exposure, with a maximum inhibition rate of 37.7%, suggesting the potential neurotoxicity of MPs to freshwater fish. The activities of cytochrome P450 (CYP) enzymes [7-ethoxyresorufin O-deethylase (EROD) and 7-benzyloxy-4-trifluoromethyl-coumarin O-dibenzyloxylase (BFCOD)] in the fish liver exhibited clear temporal variabilities, with significant decreases followed by elevations compared to the control. The alterations of the EROD and BFCOD activities indicate the potential involvement of CYP enzymes for the metabolism of MPs. The activity of antioxidative enzyme superoxide dismutase (SOD) in the liver was significantly induced throughout the exposure period, while the malondialdehyde (MDA) content did not vary with MPs exposure, suggesting that the antioxidative enzymatic system in O. niloticus could prevent oxidative damage. These results highlight the ingestion and accumulation of MPs in different tissues of freshwater fish, which lead to perturbations in fish biological systems and should be considered in environmental risk assessment.

This research aimed to evaluate the ecological risk of xenobiotics associated with agricultural activities by determining metal contents and biomarker responses using tucunaré (Cichla sp.) as a bioindicator. The work was conducted in the southwest region of the state of Tocantins, in the cities of Lagoa da Confusão and Pium. Water samples and specimens of Cichla sp. were collected in the Javaés and Formoso Rivers at three collection points (A, B and C). The concentrations of Cd, Pb, Cu, Cr, Mn, Ni and Zn in water and fish were analyzed. In fish, genotoxic, biochemical (glucose serum levels, AST (aspartate aminotransferase) and ALT (alanine aminotransferase) and histological (gills and liver) biomarkers were assessed. In the water, the Cr and Mn concentrations at the three collection points exceeded the values for Class 1 rivers. In the muscle, Cr was above the maximum limit allowed for human consumption at the three collection points, although the values at Points B and C were not significantly different from that at Point A (p > 0.05). At the three collection points, the micronucleus test revealed a low frequency of micronuclei. Significant hyperglycemia and a decrease in the AST activity of the fish collected at Point C was observed. In the gills, the most frequent alterations were at Stages I and II, which indicated mild to moderate damage, and epithelial detachment was the most frequent variation. In the liver tissue, the most frequently observed histological changes were at Stages I and II and included cytoplasmic vacuolization, nuclear hypertrophy, dilated sinusoids and bile stagnation. The integrated evaluation of these biomarkers indicated that fish collected from areas with intense agricultural activities presented adaptive responses that were likely caused by the availability and bioaccumulation of certain xenobiotics in the environment.

Selenium (Se) is an essential micronutrient for animals and humans with a relatively narrow margin between nutritional essentiality and potential toxicity. Even though our previous studies have demonstrated algae could efficiently remove Se, mainly through volatilization, concern is raised about eco-risks posed by the remaining Se in algae. Here, Sinanodonta woodiana was investigated as a biofilter for the removal of Se-containing Chlorella vulgaris and for its potential risk to human health. Our results suggest filtration rates of S. woodiana were independent of Se levels in algal biomass, with a removal efficiency of between 60 and 78%. However, Se concentrations accumulated in mussels were significantly correlated with algal-borne Se levels, with a dietary assimilation efficiency ranging from 12% to 46%. Thus, a pilot biofiltration system was set up to assess uptake and depuration processes. The system was found to efficiently remove Se laden algae through the uptake by mussels, while 21% of Se in mussels could be depurated in 6 days. Among tissues, gills accumulated the highest Se concentration after assimilating algal-borne Se but shed Se compounds in the fastest pace during depuration. Health risks posed by consumption of mussels exposed to different sources of Se were further assessed. S. woodiana accumulated the highest Se concentration after exposure to waterborne SeMet, followed by dietary Se, selenite and control. The relatively higher Se levels were found in gills for all the treatments. After boiling, the most common method of cooking mussels, the greatest reduction in Se concentration occurred in mantle for the control and dietary Se groups and in muscle for the SeMet and selenite treatments. Therefore, within the safe limits, Se-containing mussels can be consumed as a dietary supplement. Overall, our research suggests incorporation of mussels into an algal treatment system can improve Se removal efficiency and also provide financial incentives for practitioners.

This study extends our knowledge of the bioaccumulation of polycyclic aromatic hydrocarbons (PAHs) in marine organisms and investigates its possible determinants. PAH levels were measured in Solea solea tissue and in marine sediments collected from three areas of the northern Adriatic Sea characterized by different anthropic impacts (Venetian Lagoon, Po Delta, and fishing grounds off Chioggia). The possibility of differential PAH bioaccumulation in different tissues (muscle, liver and gills) was investigated by seeking relationships between mean individual and total PAH concentrations in tissue and sediment samples, the physicochemical properties of PAHs (rings and Kow), and some key biological variables (lipid content of tissues, body size, habitat). The present study demonstrated that the lipid content might not be the only determinant of PAH bioaccumulation in common sole tissues. The habitat characteristics, the tissue types and some physicochemical properties of compounds were closely related to PAH bioaccumulation.