Polychlorinated biphenyls (PCBs) are known amongst the most dangerous toxicants entering the coastal marine waters from various polluting sources. Even the smallest PCBs doses are capable to change physiological and biochemical processes exerting toxic, mutagenic and carcinogenic effects. So, the aim of this study was to analyze the impact of PCBs at 1, 100, 1000 µg/L on oxidative stress parameters (level of oxidized proteins (neutral aldehydes and ketones, basic aldehydes and ketones) and lipid peroxidation), antioxidant enzyme activities (superoxide dismutase (SOD), catalase (CAT)) and cholinesterase (ChE) activity in the hepatopancreas of mussel Mytilus galloprovincialis during 5 days’ of the toxicological experiment. Level of all forms of the oxidized proteins was found significantly increased at 100 µg/L and 1000 µg/L (+50-78% and +150-282%, respectively) compared to the control (p<0.05). Level of lipid peroxidation was considerably higher at 1, 100, 1000 µg/L (+59%, +134%, +269%, respectively) compared to the control (p<0.05). SOD activity significantly raised at 1, 100, 1000 µg/L (+63%, +200%, +118%, respectively) compared to the control (p<0.05), while CAT activity reduced at 1000 µg/L compared to the control, 1 µg/L and 100 µg/L (-29%, -66%, -40%, respectively) (p<0.05). ChE activity was found lower at 1, 100, 1000 µg/L (-60%, -93%, -30%, respectively) compared to the control (p<0.05). Possible mechanisms of M. galloprovincialis biochemical response to PCBs are discussed. Studied biochemical parameters can be suitable biomarkers for evaluating the toxicity of PCBs and M. galloprovincialis can be used as a bioindicator in the monitoring of marine ecosystems contaminated with these pollutants.

Metal-based nanoparticles (NPs) are considered detrimental to aquatic organisms due to their potential accumulation. However, little is known about the mechanisms underlying these effects and their species-specificity. Here we used stable silver (Ag) NPs (20 nm, from 10 to 500 μg/L) with a low dissolution rate (≤2.4%) to study the bioaccumulation and biological impacts in two freshwater gastropods: Lymnaea stagnalis and Planorbarius corneus. No mortality was detected during the experiments. Ag bioaccumulation showed a dose-related increase with an enhanced concentration in both species after 7d exposure. L. stagnalis displayed a higher accumulation for AgNPs than P. corneus (e.g., up to 18- and 15-fold in hepatopancreas and hemolymph, respectively) which could be due to the more active L. stagnalis having greater contact with suspended AgNPs. Furthermore, the hepatopancreas and stomach were preferred organs for bioaccumulation compared to the kidney, mantle and foot. Regarding biological responses, the hemolymph rather than hepatopancreas appeared more susceptible to oxidative stress elicited by AgNPs, as shown by significantly increasing lipid peroxidation (i.e., formation of malondialdehyde). Neurotoxicity was detected in L. stagnalis when exposed to high concentrations (500 μg/L). Comparison with impacts elicited by dissolved Ag revealed that the effects observed on AgNPs exposure were mainly attributable to NPs. These results highlighted the relationship between the physiological traits, bioaccumulation, and toxicity responses of these two species to AgNPs and demonstrated the necessity of species-specificity considerations when assessing the toxicity of NPs.

This study reports the shapes, dimensional classes, types and counts of microplastics (MPs) found in 23 individuals of N. norvegicus collected from two wild populations of the Adriatic Sea (Mediterranean basin). The focus was on three different anatomical compartments (gut, hepatopancreas and tail), which were analysed separately. MPs were found in all the investigated individuals with an average of about 17 MPs/individual. Fragments were predominant over fibers with a ratio of about 3:1. The majority of MPs were in the dimensional range 50–100 μm. The predominant polymers were polyester, polyamide 6, polyvinyl chloride and polyethylene, which together constitute about 61% of all the MPs found. Fragments were more concentrated in the hepatopancreas, with no significant difference between gut and tail, while fibers were more concentrated in the gut than in the tail with hepatopancreas somehow in between. The dimensional class of the MPs influences their anatomical distribution. There were no statistical differences among individuals from the two sampling sites. Sex of the individual did not influence the level of retained MPs, while length had a very marginal effect. The information reported here contributes to understanding of the possible risks linked to human consumption of different tissues from contaminated Norway lobsters.

Cadmium (Cd) is being frequently detected in marine organisms. However, dose-dependent effects of Cd challenged unraveling the toxicological mechanisms of Cd to marine organisms and developing biomarkers. Here, the dose-dependent effects of Cd on clams Ruditapes philippinarum following exposure to 5 doses of Cd (3, 9, 27, 81, 243 μg/L) were investigated using benchmark dose (BMD) method. By model fitting, calculation of BMD values was performed on transcriptomic profiles, metals concentrations, and antioxidant indices. Cd exposure induced not only significant Cd accumulation in clams, but also marked alterations of essential metals such as Ca, Cu, Zn, Mn, and Fe. Gene regulation posed little influence on essential metal homeostasis, indicated by poor enrichment of differentially expressed genes (DEGs) associated with metal binding and metal transport in lower concentrations of Cd-treated groups. BMD analysis on biological processes and pathways showed that peptide cross-linking was the most sensitive biological process to Cd exposure, followed by focal adhesion, ubiquitin mediated proteolysis, and apoptosis. Occurrence of apoptosis was also confirmed by TUENL-positive staining in gills and hepatopancreas of clams treated with Cd. Furthermore, many DEGs, such as transglutaminases (TGs), metallothionein (MT), STEAP2-like and laccase, which presented linear or monotonic curves and relatively low BMD values, were potentially preferable biomarkers in clams to Cd. Overall, BMD analysis on transcriptomic profiles, metals concentrations and biochemical endpoints unraveled the sensitiveness of key events in response to Cd treatments, which provided new insights in exploring the toxicological mechanisms of Cd in clams as well as biomarker selection.

The present paper reports on the interrelationships of fish, parasites and the bioaccumulation of hazardous organic compounds in the Zemplínska Šírava water reservoir in eastern Slovakia, which is heavily polluted with polychlorinated biphenyls (PCBs). The concentrations of these contaminants were measured in various fish matrices (dorsal and abdominal muscle tissues, hepatopancreas, intestine wall and adipose tissue) of the freshwater bream, Abramis brama (Cyprinidae), and in its intestinal parasite Caryophyllaeus laticeps (Cestoda), which was used for the first time as a model for a PCB bioaccumulation study. Regarding the fish, the highest concentrations of PCBs were found in the intestine, followed by hepatopancreas and muscle tissues. The amounts of PCBs were higher in abdominal muscles than in their dorsal parts. Concentrations of ∑PCBs above the limits set by European regulations were detected in both muscle parts in the fish, confirming the persistent unfavorable conditions in this locality and high risk for biota and humans. Based on bioconcentration factor values (BCFs), PCBs reached much higher levels in cestodes compared to bream matrices. Some significant differences in PCB amounts between infected and uninfected bream were determined. Fulton's condition factor (CF) significantly differed in infected and non-infected fish (p ˂ 0.05), with CF values surprisingly lower in fish free of parasites compared to parasitized fish, which suggests a “mutualistic” relationship between the parasite and its host.

Various crustaceans are farmed using aquaculture, and food deprivation or fasting can occur due to changing of environmental or management strategies. However, the molecular mechanisms underlying responses to starvation in crustaceans remain unclear. To address this, 12 hepatopancreas transcriptomes were compared for oriental river prawn (Macrobrachium nipponense) from four fasting stages (0, 7, 14 and 21 d). Gene Ontology functional annotation and Kyoto Encyclopaedia of Genes and Genomes pathway enrichment analysis of differentially expressed genes were subsequently performed. During the early stages of starvation (0–7 d), drug metabolism via the cytochrome P450 pathway and metabolism of xenobiotics by the cytochrome P450 pathway were enriched, suggesting that they metabolised compounds generated under starvation stress. As starvation proceeded (7–14 d), the retinol (vitamin A) metabolism pathway was activated, based on three up-regulated genes (CYP3, ADH and UGT), along with the two p450 pathways. Meanwhile, vitamin A was gradually consumed. As acute starvation was reached (14–21 d), vitamin A deficiency decreased the mRNA expression levels of IGF-I that is involved in the mTOR signalling pathway, which ultimately affected the growth and development of M. nipponense. Our results implicate drug/xenobiotic metabolism by cytochrome P450s in adaptation to starvation stress. Furthermore, metabolic cascades (CYP and retinol pathways) and growth (mTOR signalling) pathways are clearly triggered in crustaceans during starvation. The findings expand our understanding of the genes associated with hepatopancreas functioning in M. nipponense, and the underlying molecular mechanisms that govern the responses of crustaceans to starvation stress.

Blue crabs from a contaminated estuary (Hackensack Meadowlands-HM) and a cleaner reference site (Tuckerton-TK) were analyzed for Cr, Cu, Hg, Pb, and Zn in muscle and hepatopancreas. Crabs from each site were taken into the laboratory and fed food from the other site, or in another experiment, transplanted to the other site for eight weeks. All crabs were analyzed for metals. Overall, tissue concentrations reflected environmental conditions. Tissue differences were found for Cu, Pb and Zn (all higher in hepatopancreas), and Hg (higher in muscle). HM muscle had more Hg than TK muscle, but did not decrease after transplanting or consuming clean food. HM crabs lost Cu, Pb and Zn in hepatopancreas after being fed clean food or transplanted. TK crabs increased Hg in muscle and Cr and Zn in hepatopancreas after transplantation or being fed contaminated (HM) food. Concentrations were variable, suggesting that blue crabs may not be fully reliable bioindicators of polluted systems. The accumulation of metals within the muscle and hepatopancreas of blue crabs was highly variable, but often followed environmental concentrations.

The disruption of the Fundão dam released 43 million m³ of mine tailings into the Doce River until it flowed into the ocean through the estuary. The mine tailing changed the composition of metals in water and sediment, creating a challenging scenario for the local biota. We used multivariate analyzes and the integrated biomarker response index (IBR) to assess the impact of mine tailings on the bioaccumulation profile (As, Cd, Cr, Cu, Fe, Mn, Pb and Zn) as well as the biomarkers response in gills, hepatopancreas and muscle of shrimps sampled from different sectors during two dry seasons (dry1 and dry2) (Sep/Oct 2018; 2019) and two wet seasons (wet1 and wet2) (Jan/feb 2019; 2020). There was seasonal and local effect under bioaccumulation and biomarker response revealing that the pattern responses seen in each sector sampled changed according to the season. The greater IBR added to the strong association among the most metals tissue content (Cd, Cr, Cu and Mn) and sectors sampled during dry 1 suggests greater bioavailability of these metals to the environment in this period. Estuarine sectors stand out for high Fe bioavailability, especially during wet1, which seems to be associated with greater metallothionein content in hepatopancreas of shrimps. Native species of marine shrimps proved to be successful indicators of sediment quality besides being sensitive to water contamination by metals. The multi-biomarkers approach added to multivariate analysis supports the temporal and seasonal effects, signalizing the importance of continuous monitoring of the estuarine region to better know about the bioavailability of these metals, mainly Fe, and their long-term effects on the local biota.

Cadmium (Cd) is a toxic heavy metal that can be discharged into water environment through industrial activities, threatening the health of aquatic organisms and humans. MicroRNA (miRNA) plays an important role in the process of autophagy. The purpose of this experiment was to study the mechanism of Cd-induced autophagy in common carp hepatopancreas. We established a Cd poisoning model of common carp and explored ultrastructure, two oxidation indicators, three antioxidant indicators, miR-25-3p, two heat shock proteins (Hsps), and nine autophagy-related genes. The results confirmed that deleterious effect of Cd caused the injury of hepatopancreas and the appearance of hepatopancreas autophagic cells in common carp. At the same time, Cd exposure increased the contents of hydrogen peroxide (H₂O₂) and malonaldehyde (MDA), and decreased the activities of catalase (CAT), superoxide dismutase (SOD), and total antioxidative capacity (T-AOC), meaning that Cd caused oxidative stress via the imbalance between peroxide level and antioxidant capacity. Moreover, exposure to Cd increased mRNA expression of microtubule associated protein-1 light chain 3 beta (LC3-II), Dynein, Beclin 1, autophagy-related gene 5 (Atg5), and autophagy-related gene 12 (Atg12); and decreased mRNA expression of mechanistic target of rapamycin kinase (mTOR), indicating that excess Cd caused autophagy, and AMPK/mTOR/ULK1 signaling pathway took part in autophagy induced by Cd in common carp hepatopancreas. Furthermore, Cd down-regulated miR-25-3p and up-regulated its three target genes (AMPK, ULK1 as well as PTEN), suggesting that miR-25-3p mediated autophagy induced by Cd. In addition, we found that Hsps were activated via the up-regulation of Hsp70 and Hsp90. Moreover, oxidative stress mediated autophagy via Hsps in Cd-treated common carp hepatopancreas and Cd-induced autophagy was time dependent. In summary, miR-25-3p, oxidative stress, and Hsps participated in autophagy caused by Cd in common carp hepatopancreas. This study provided a new idea for the mechanism of Cd-induced autophagy in hepatopancreas.

DCOIT (4,5-dichloro-2-n-octyl-4-isothiazolin-3-one) is the main component of SeaNine-211, a new antifouling agent that replaces tributyltin to prevent the growth of undesirable organisms on ships. There have been some studies on the toxicity of DCOIT, but the mechanism of DCOIT’s toxicity to crustaceans still requires elucidation. This study examined the chronic toxicity (4 weeks) of 0, 3, 15, and 30 μg/L DCOIT to the Pacific white shrimp (Litopenaeus vannamei) from the aspects of growth and physiological and histological changes in the hepatopancreas and gills. A transcriptomic analysis was performed on the hepatopancreas to reveal the underlying mechanism of DCOIT in shrimp. The exposure to 30 μg/L DCOIT significantly reduced the survival and weight gain of L. vannamei. High Na⁺/K⁺-ATPase activity and melanin deposition were found in the gills after 4 weeks of 15 μg/L or 30 μg/L DCOIT exposure. The highest concentration of DCOIT (30 μg/L) induced changes in hepatopancreatic morphology and metabolism, including high anaerobic respiration and the accumulation of triglycerides. Compared with the exposure to 3 μg/L DCOIT, shrimp exposed to 15 μg/L DCOIT showed more differentially expressed genes (DEGs) than those in the control, and these DEGs were involved in biological processes such as starch and sucrose metabolism and choline metabolism in cancer. The findings of this study indicate that L. vannamei is sensitive to the antifouling agent DCOIT and that DCOIT can induce altered gene expression at a concentration of 15 μg/L and can interfere with shrimp metabolism, growth and survival at 30 μg/L.