Hepatopancreas Research Articles

Potential toxic effects of titanium oxide (TiO2) nanoparticles on the biological, biochemical, and histological aspects of the land snail Helix aspersa

Nanotechnology has come a long way in our lives. However, it maintains some negative effects on the environment. This study aims to use the land snail Helix aspersa as a bioindicator. Titanium dioxide nanoparticles (TiO2NPs) had been used at 70 and 140 µg/L for two weeks by the spraying method. The oxidative biomarkers, condition index (CI), DNA damage, hemocyte count, and phagocytic activity were estimated. The toxicity of TiO2NPs was determined (LC50 = 544 µg/L). The exposure to TiO2NPs caused a significant reduction of the activities of superoxide dismutase (SOD) and catalase (CAT) in the digestive gland of Helix aspersa (the activity of CAT was 3.4 ± 0.1 (P = 0.001), SOD was 11 ± 1 (P = 0.0002) at concentration 140 µg/L after two weeks). The activity of glutathione peroxidase (GPX) was (1.13 ± 0.01 µ/mg protein at 140 µg/L compared with controls (5.47 ± 0.01 µ/mg protein). The treatment caused DNA damage in the hemocytes (tail DNA % = 8.66 ± 0.02 and tail moment = 52.99 ± 0 at140 µg/L (P = 0.002)). In the digestive gland, both tail DNA % and tail moment increased (tail moment = 78.38 ± 0.08 compared with control = 2.29 ± 0.09 (P = 0.0001)). The total count of hemocytes significantly decreased after two weeks (the average number was 71 ± 1.5 compared with controls 79 ± 1.1 at 140 µg/L). Furthermore, TiO2NPs caused histological alterations in the digestive gland of Helix aspersa. It can be concluded that the Helix aspersa can be used as environmental pollution bioindicator. A comprehensive evaluation of toxic effects induced by TiO2NPs in vivo assays must be investigated.

Comparative toxicity of micro and nanopolystyrene particles in Mya arenaria clams

The contamination of coastal marine environments by plastics of sizes ranging from mm down to the nanoscale (nm) could pose a threat to aquatic organisms. The purpose of this study was to examine the toxicity of polystyrene nanoparticles (PsNP) of various sizes (50, 100 and 1000 nm) to the marine clams Mya arenaria. Clams were exposed to concentrations of PsPP for 7 days at 15 °C and analyzed for uptake/transformation, changes in energy metabolism, oxidative stress, genotoxicity and circadian neural activity. The results revealed that PsNP accumulated in the digestive gland was 50 nm > 100 nm > 1000 nm. All sized increased oxidative stress as follows: 50 nm (peroxidase, antioxidant potential and LPO), 100 nm (LPO and antioxidant potential) and 1000 nm (LPO). Tissue damage was also size dependent by increasing genotoxicity. The 100 nm PsPP altered the levels of the circadian metabolite melatonin. We conclude that the toxicity of plastics is size dependent in clams.

Variation of Gene Expression in the Endemic Dinaric Karst Cave-Dwelling Bivalve Mollusk Congeria kusceri during the Summer Season

The cave systems of the Neretva River basin in the Dinaric Karst are home to Congeria kusceri, one of the very few known examples of stygobiotic bivalve mollusks, which displays several unique life history traits and adaptations that allowed its adaptation to the subterranean environment. This endemic species is undergoing rapid decline, most likely linked with habitat degradation, which might seriously threaten its survival in the next few decades. Unfortunately, the urgent need for effective conservation efforts is hampered by the lack of effective regulations aimed at preserving remnant populations as well as by our limited knowledge of the biology of this species. Although the precise factors underlying the disappearance of C. kusceri from its type locations are not entirely clear, the alteration of seasonal changes in water temperatures and alkalinity is most likely involved, as these are the main drivers of shell growth, spawning, and the onset and progression of the new gametogenic cycle. While these aspects have so far only been studied using morphological and physiological observations, future conservation efforts would certainly benefit from an integrated multidisciplinary approach. Here, using RNA-sequencing, we provide an overview of the modulation of gene expression recorded in five key tissues (the mantle, gonads, gills, adductor muscle, and digestive gland) between early June and late September. We highlight the presence of markedly tissue-specific responses, with the most dramatic changes affecting the gonads, mantle, and gills. We further identified a drastic switch in the use of energy budgets between the two periods, with evidence of ongoing shell growth and high metabolic activity in the mantle and gills at the early time point, followed by a massive redirection of all available energy to the gonads for the generation of new gamete primordia in early autumn.

Fitness assessment of Mytilus galloprovincialis Lamarck, 1819 after exposure to herbicide metabolite propachlor ESA

The lack of data on the chronic effects of chloroacetanilide herbicide metabolites on non-target aquatic organisms creates a gap in knowledge about the comprehensive impacts of excessive and repeated pesticide use. Therefore, this study evaluates the long-term effects of propachlor ethanolic sulfonic acid (PROP-ESA) after 10 (T1) and 20 (T2) days at the environmental level of 3.5 μg.L−1 (E1) and its 10x fold multiply 35 μg.L−1 (E2) on a model organism Mytilus galloprovincialis. To this end, the effects of PROP-ESA usually showed a time- and dose-dependent trend, especially in its amount in soft mussel tissue. The bioconcentration factor increased from T1 to T2 in both exposure groups – from 2.12 to 5.30 in E1 and 2.32 to 5.48 in E2. Biochemical haemolymph profile and haemocyte viability were not affected by PROP-ESA exposure. In addition, the viability of digestive gland (DG) cells decreased only in E2 compared to control and E1 after T1. Moreover, malondialdehyde levels increased in E2 after T1 in gills, and DG, superoxidase dismutase activity and oxidatively modified proteins were not affected by PROP-ESA. Histopathological observation showed several damages to gills (e.g., increased vacuolation, over-production of mucus, loss of cilia) and DG (e.g., growing haemocyte trend infiltrations, alterations of tubules). This study revealed a potential risk of chloroacetanilide herbicide, propachlor, via its primary metabolite in the Bivalve bioindicator species M. galloprovincialis. Furthermore, considering the possibility of the biomagnification effect, the most prominent threat poses the ability of PROP-ESA to be accumulated in edible mussel tissues. Therefore, future research about the toxicity of pesticide metabolites alone or their mixtures is needed to gain comprehensive results about their impacts on living non-target organisms.

Transcriptome Analysis Reveals the Genes Involved in Oxidative Stress Responses of Scallop to PST-Producing Algae and a Candidate Biomarker for PST Monitoring.

Paralytic shellfish toxins (PST) could be accumulated in bivalves and cause safety problems. To protect public health, bivalves are examined for PST contamination before entering the market, usually by high-performance liquid chromatography (HPLC) or LC-tandem mass spectrometry (LC-MS/MS) in the lab, which needs PST standards not all available and is time-consuming for large sample sizes. To detect PST toxicity in bivalves rapidly and sensitively, a biomarker gene is highly demanded, but the related study is very limited. In this study, we fed a commercially important bivalve, Patinopecten yessoensis, with the PST-producing dinoflagellate Alexandrium catenella. After 1, 3, and 5 days of exposure, both PST concentrations and toxicity levels in the digestive gland continuously increased. Transcriptome analysis revealed that the differentially expressed genes were significantly enriched in oxidation-reduction process, which included the cytochrome P450 genes (CYPs), type I iodothyronine deiodinase (IOD1s), peroxidasin (PXDN), and acyl-Coenzyme A oxidase 1 (ACOX1) at day 1 and a superoxide dismutase (SOD) at day 5, highlighting the crucial roles of these genes in response to oxidative stress induced by PST. Among the 33 continuously upregulated genes, five showed a significant correlation between gene expression and PST concentration, with the highest correlation present in PyC1QL4-1, the gene encoding Complement C1Q-like protein 4, C1QL4. In addition, the correlation between PyC1QL4-1 expression and PST toxicity was also the highest. Further analysis in another aquaculture scallop (Chlamys farreri) indicated that the expression of CfC1QL4-1, the homolog of PyC1QL4-1, also exhibited significant correlations with both PST toxicity and concentration. Our results reveal the gene expression responses of scallop digestive glands to PST-producing algae and indicate that the C1QL4-1 gene might be a potential biomarker for PST monitoring in scallops, which may provide a convenient way for the early warning and sensitive detection of PST contamination in the bivalves.

The Antarctic Scallop Adamussium colbecki Is Unable to Transcriptomically Respond to Captivity and Moderate Thermal Stress

Adamussium colbecki is a scallop endemic of the Antarctic Ocean, the only modern survivor of the Adamussiini tribe and one of the few bivalves living in polar environments. Compared with other Antarctic animals, very little is known concerning the evolutionary adaptations which allow this species to thrive at sub-zero temperatures. Due to its local abundance and sensitivity to environmental changes, A. colbecki is an interesting model for studying the effects of pollution and climate change in the Antarctic Ocean. Here, we report, for the first time, the application of transcriptomic tools to the study of the effects of a short-to-medium term exposure to a +1.5 °C water temperature increase on three tissues. Although this approach did not highlight any significant change in response to thermal stress, we observed slight alterations in energetic metabolism and nutrient adsorption in the digestive gland, most likely linked with stabling in experimental tanks. The results of our study suggest that A. colbecki may be particularly vulnerable to the effects of climate change due to its complete inability to adapt to temperature increase at the transcriptomic level.

Multiple responses of Mytilus galloprovincialis to plastic microfibers

Plastic microfibers (MF) represent the major source of MF found in the environment, the majority made of polyester (PES). Marine bivalves, suspension feeders widespread in coastal areas subjected to higher anthropogenic input, can accumulate MF from the water column in their tissues. This raised some concern about their possible impact on bivalve health and potential transfer along the food chain. In this work, the effects of PES-MF on the mussel Mytilus galloprovincialis were investigated, utilizing MF obtained by cryo-milling of a fleece cover. Fiber characterization indicated the polymer composition as polyethylene terephthalate (PET); the size distribution was in a length range resembling that of MF released from textile washing, and including those that can be ingested by mussels. MF were first screened for short-term in vitro immune responses in mussel hemocytes. The effects of in vivo exposure (96 h, 10 and 100 μg/L, corresponding to about 150 and 1500 MF/mussel/L, respectively), were then evaluated. Data are presented on hemolymph immune biomarkers (Reactive Oxygen Species and nitric oxide production, lysozyme activity), and on antioxidant biomarkers (catalase and glutathione S-transferase) and histopathology in gills and digestive gland. Tissue MF accumulation was also evaluated.MF exposure stimulated extracellular immune responses both in vitro and in vivo, indicating induction of immune/inflammatory processes. In both tissues, stimulation of antioxidant enzyme activities, suggesting oxidative stress conditions, and histopathological changes were observed, with stronger effects often observed at lower concentration. Although mussel retained a very small fraction of MF, their accumulation was higher in the digestive gland than in gills, and in both tissues of mussels exposed to the lowest concentration. Selective accumulation of shorter MF was also observed, particularly in gills. Overall, the results demonstrate that at environmental exposure levels, PET-MF have a significant impact on mussel physiology, affecting multiple processes in different tissues.

Duplicated paralog of sulfide: quinone oxidoreductase contributes to the adaptation to hydrogen sulfide-rich environment in the hydrothermal vent crab, Xenograpsus testudinatus

The hydrothermal crab, Xenograpsus testudinatus (xtcrab) inhabits shallow-water, hydrogen sulfide (H2S)-rich hydrothermal vent regions. Until now, the adaptative strategy of xtcrab to this toxic environment was unknown. Herein, we investigated the sulfide tolerance and detoxification mechanisms of xtcrabs collected in their high-sulfide hydrothermal vent habitat. Experimental immersion of xtcrab in various sulfide concentrations in the field or in aquaria assessed its high sulfide tolerance. HPLC measurement of hemolymph sulfur compounds highlighted xtcrab detoxification capacity via catabolism of sulfide into much less toxic thiosulfate. We focused on a key enzyme for H2S detoxification, sulfide: quinone oxidoreductase (SQR). Cloning and phylogenetic analysis revealed two SQR paralogs in xtcrab, that we named xtSQR1 and xtSQR2. As shown by qPCR, xtSQR2 and xtSQR1 were expressed in the digestive gland, suggesting the involvement of both paralogs in the detoxification of food-related H2S. In contrast, xtSQR1 transcript was highly expressed in the gill, while xtSQR2 was not detectable, suggesting a specific role of SQR1 in gill detoxification of H2S of environmental origin. Comparison between xtcrabs in their hydrogen sulfide-rich hydrothermal habitat, and xtcrabs maintained for one month in sulfide-free seawater aquarium, showed higher transcript levels of gill xtSQR1 in sulfide-rich habitat, further supporting the specific role of xtSQR1 paralog in environmental H2S detoxification in the gill. Gill SQR protein level as measured by Western blot, and gill SQR enzyme activity were also higher in sulfide-rich habitat. Immunohistochemical staining further showed that SQR expression was co-localized with Na+/K+-ATPase-positive epithelial and pillar cells of the gill filament. This is the first evidence of duplicate SQR genes in crustaceans. Overall, our study suggests that the subfunctionalization of duplicate xtSQR genes may play an important role in sulfide detoxification to maintain the sulfide homeostasis in X. testudinatus, providing an ecophysiological basis for its adaptation to the high-sulfide hydrothermal vent environment.

Physiological and biochemical responses to caffeine and microplastics in Mytilus galloprovincialis

Caffeine (Caff) is one of the most widely used substances in the human diet and a well-recognized drug. Its input into surface waters is remarkable, but biological effects on aquatic organisms are unclear, particularly in combination with pollutants of suspected modulatory activity, like microplastics (MP). The aim of this study was to reveal the impact on the marine mussel Mytilus galloprovincialis (Lamark, 1819) of Caff (20.0 μg L−1) in the environmentally relevant combination (Mix) with MP 1 mg L−1 (size 35–50 μm) after the exposure for 14 days. Untreated and exposed to Caff and MP separately groups were also examined. Cell viability and cell volume regulation in hemocytes and digestive cells, as well as the indexes of oxidative stress, glutathione (GSH/GSSG) and metallothioneins levels, and caspase-3 activity in digestive gland were assessed. MP and Mix reduced Mn-superoxide dismutase, catalase, and glutathione S-transferase activities and level of lipid peroxidation, but increased the digestive gland cell viability, GSH/GSSG ratio (by 1.4–1.5-fold), metallothioneins level and their Zn content, while Caff did not affect oxidative stress indexes and metallothionein-related Zn chelation. Protein carbonyls were not targeted in all exposures. The distinguishing feature of the Caff group was the decline (2-fold) in caspase-3 activity and low cell viability. The multi-stress effect of Mix was shown by the worsening of the volume regulation of digestive cells and confirmed by discriminant analysis of biochemical indexes. The special capabilities of M. galloprovincialis as a sentinel organism make it an excellent bio-indicator reflecting the multi-stress effects in sub-chronic exposures to potentially harmful substances. The identification of the modulation of individual effects in combined exposure increases the need to base monitoring programs on studies of multi-stress effects in sub-chronic exposures.

Biochemical and histopathological effects of copper oxide nanoparticles exposure on the bivalve Chambardia rubens (Lamarck, 1819).

Copper nanoparticles are widely incorporated into many applications, including air and liquid filters, wood preservatives, batteries, thermal and electrical conductivity, inks and skin products. Their potential toxicity and environmental fate, however, are poorly studied in the freshwater bivalves. The aim of the present study was to evaluate the different effects of copper oxide nanoparticles and ionic copper on the digestive glands and gills of the mussel Chambardia rubens. Mussels were treated with 100 and 1000 µg Cu L-1 of copper oxide nanoparticles (CuONPs) or ionic copper (Cu2+) for 3, 7, and 14 days. The Cu accumulation and markers of oxidative stress in the digestive glands and gills were evaluated. The results show that the digestive gland collected higher levels of the two forms of copper than the gills. Exposure to CuONPs or Cu2+ induced significant elevations in superoxide dismutase, glutathione peroxidase and lipid peroxidation. Notably, a significant decrease was observed in the glutathione levels after exposure to both copper forms. CuONPs only induced a significant increase in glutathione reductase and glutathione S-transferase. The ionic copper only induced a significant decrease in catalase activities in the gill tissues. Overall, CuONPs and Cu2+ provoked oxidative stress, and further research is needed to clarify their genotoxic and neurotoxic effects on freshwater mussels and other biota.

Determination of gonad reproductive state using non-lethal ultrasonography in endangered black (Haliotis cracherodii) and white abalone (H. sorenseni)

IntroductionBlack (H. cracherodii) and white abalone (H. sorenseni) are federally listed as endangered species in the United States. Conservation efforts include captive breeding programs; however, determination of the reproductive state of individual abalone is notoriously difficult using traditional visual assessments. Ultrasonography is a well-recognized technology used to assess gonad reproductive condition accurately and nonlethally in cultured and wild fish, and more recently cultured red abalone (H. rufescens). Here, we implemented the use of ultrasound imaging technology to monitor the gonad condition of endangered black and white abalone.MethodsRepeated ultrasound assessments of the gonad were used to assess seasonal changes in reproductive development in wild black (n=20), and captive white abalone (n=25).ResultsA modified ultrasound gonad index score was developed to incorporate multiple species of abalone. The ultrasound index scores ranged from one to five, with an index score of one being the lowest (gonad margin is thinly wrapped around the digestive gland or not present) and an index score of five being the highest (gonad margin is thick and significantly compressing the digestive gland).ConclusionWe show that non-lethal ultrasound imaging technology is useful for tracking cyclical changes in the gonad reproductive condition as well as a more precise selection of individuals that are in peak reproductive condition for captive breeding programs.

Comparative biochemical and molecular responses of biotransformation and antioxidant systems in three species of Crassostrea (Sacco, 1897) oysters exposed to chrysene

Chrysene (CHR) is among the most persistent polycyclic aromatic hydrocarbons (PAH) in water and a priority compound for pollutants monitoring, due to its carcinogenic, mutagenic and genotoxic potential. Aquatic animals exposed to CHR may present alterations of biomarkers involved in the biotransformation and oxidative stress-related parameters. The aim of this study was to investigate differences in antioxidant and biotransformation (phase I and II) systems of Crassostrea gigas, C. gasar and C. rhizophorae and its effects resulting from CHR exposure. Adult oysters of these species were exposed to 10 μg L−1 of CHR for 24 h and 96 h. In gills, the transcripts CYP1-like, CYP2-like, CYP2AU1-like, GSTO-like, MGST-like, SULT-like were evaluated after 24 h of exposure. The activity of SOD, CAT, GPx, GR and G6PDH were analyzed in gills and digestive glands after 96 h of exposure. CHR bioaccumulated in tissues. Differences in the remaining levels of CHR in water after 96 h were observed in aquaria containing C. gigas or C. gasar oysters and may be associated to the different filtration rates between these species. Downregulate of biotransformation genes were observed in gills of C. gasar (CYP2AU1-like and GSTO-like) and C. rhizophorae (CYP1-like1, CYP2-like, MGST-like and SULT-like), suggesting that biotransformation responses may be species-specific. Differential activity of antioxidant enzymes were observed in gills and digestive gland of oysters exposed to CHR. Biochemical responses suggested that C. gigas and C. gasar are more responsive to CHR. Differential responses observed among the three Crassostrea species can be related to evolutionary differences, ecological niches and adaptation to environment.

Seasonal dynamics of the microbiome-host response to pharmaceuticals and pesticides in Mytilus galloprovincialis farmed in the Northwestern Adriatic Sea

Marine mussels, especially Mytilus galloprovincialis, are well-established sentinel species, being naturally resistant to the exposure to multiple xenobiotics of natural and anthropogenic origin. Even if the response to multiple xenobiotic exposure is well known at the host level, the role of the mussel-associated microbiome in the animal response to environmental pollution is poorly explored, despite its potential in xenobiotic detoxification and its important role in host development, protection, and adaptation. Here, we characterized the microbiome-host integrative response of M. galloprovincialis in a real-world setting, involving exposure to a complex pattern of emerging pollutants, as occurs in the Northwestern Adriatic Sea. A total of 387 mussel individuals from 3 commercial farms, spanning about 200 km along the Northwestern Adriatic coast, and in 3 different seasons, were collected. Multiresidue analysis (for quantitative xenobiotic determination), transcriptomics (for host physiological response), and metagenomics (for host-associated microbial taxonomical and functional features) analyses were performed on the digestive glands. According to our findings, M. galloprovincialis responds to the presence of the complex pattern of multiple emerging pollutants – including the antibiotics sulfamethoxazole, erythromycin, and tetracycline, the herbicides atrazine and metolachlor, and the insecticide N,N-diethyl-m-toluamide – integrating host defense mechanisms, e.g., through upregulation of transcripts involved in animal metabolic activity, and microbiome-mediated detoxification functions, including microbial functionalities involved in multidrug or tetracycline resistance. Overall, our data highlight the importance of the mussel-associated microbiome as a strategic player for the orchestration of resistance to the multixenobiotic exposure at the holobiont level, providing strategic functionalities for the detoxification of multiple xenobiotic substances, as occurring in real world exposure settings. Complementing the host with microbiome-dependent xenobiotic degradative and resistance genes, the M. galloprovincialis digestive gland associated microbiome can have an important role in the detoxification of emerging pollutants in a context of high anthropogenic pressure, supporting the relevance of mussel systems as potential animal-based bioremediation tool.

Biochemical and behavioral effects of zinc oxide nanorods on the freshwater mussel Potomida littoralis

ABSTRACT Our study aimed to investigate the impact of nano-zinc oxide (nZnO), a widely used pollutant in industry, pharmaceuticals, and personal care products, on the behavior and oxidative stress of freshwater mussels (Potomida littoralis) an indicator species and also a model non-target organism in ecotoxicology. To this end mussels were exposed to nZnO (50 and 100 µg/L) and Zn2+ from ZnSO4 (50 and 100 µg/L) for 7 days. ZnSO4 was used for comparison purposes and to determine if the toxicity of nZnO was due to the release of ions into the aquatic environment. We evaluated changes in oxidative stress markers, including catalase (CAT), glutathione-S-transferase (GST), acetylcholinesterase (AChE), and malondialdehyde (MDA) levels, on the mussel gills and digestive glands. Additionally, the effect of nZnO on the filtration rate of bivalves was studied. The findings showed that the mussel tissue’s different parameters were significantly affected by exposure to various concentrations of nZnO, causing changes in their behavior that led to a decrease in filtration rate. Additionally, noteworthy increments were observed in CAT activity, AChE activity, and MDA levels, whereas GST activity displayed a decreasing trend, implying that oxidative stress contributes to the toxicity of nZnO. The purpose of this review is to present a framework for comprehending the toxicological impacts of nanoparticles from an environmental standpoint. Additionally, it includes novel information about the connections between nanoparticles (NPs) and bivalve species.

Evidences of metabolic alterations and cellular damage in different tissues of scallops Aequipecten tehuelchus exposed to cadmium

Scallops Aequipecten tehuelchus (Patagonia, Argentina) were exposed to 0, 2, 5 and 12 μg Cd/L for 7 and 14 days, causing in digestive gland a significant production of reactive oxygen and nitrogen species (RONS), induction of catalase (CAT) and glutathione S-transferase (GST) activities and metallothioneins (MT) synthesis. In gills, there was inhibition of GST and induction of CAT, MT and α-tocopherol (α-Toc). In muscle, a significant increment of MT was also registered and inhibition of CAT. Lipid peroxidation, measured as TBARS, was not promoted in any tissue. More significant effects were observed in digestive gland than in gills and muscle, evidencing the critical role of digestive gland in Cd accumulation and metabolisation. This research would evidence dose-dependent effects of Cd on MT, GST, CAT and α-Toc in the three organs assayed, as well as a time-dependent effect of Cd on the response of CAT, GST and TBARS in digestive gland.

Portunus pelagicus (Linnaeus, 1758) as a Sentinel Species to Assess Trace Metal Occurrence: A Case Study of Kuwait Waters (Northwestern Arabian Gulf).

Heavy metal pollution can adversely impact marine life, such as crabs, which can accumulate it in different organs and potentially transfer and biomagnify along the food chain in aquatic ecosystems. This study aimed to examine the concentrations of heavy metals (Cd, Cu, Pb, and Zn) in sediment, water, and crab tissues (gills, hepatopancreas, and carapace) of the blue swimmer crab Portunus pelagicus in the coastal areas of Kuwait, northwestern Arabian Gulf. Samples were collected from Shuwaikh Port, Shuaiba Port, and Al-Khiran areas. The accumulation of metals in crabs were higher in the carapace > gill > digestive gland, and the highest metal concentration was found in crabs collected from Shuwaikh > Shuaiba > Al-Khiran. The metal concentrations in the sediments were in the order Zn > Cu > Pb > Cd. Zn was the highest metal concentration detected in marine water sampled from the Al-Khiran Area, whereas the lowest metal was Cd sampled in water from the Shuwaikh Area. The results of this study validate the marine crab P. pelagicus as a relevant sentinel and prospective bioindicator for evaluating heavy metal pollution in marine ecosystems.

Content of Trace Elements in Tissues of Gastropod Mollusks of the Family Buccinidae (Gastropoda) off the Coasts of South-Eastern Sakhalin (the Sea of Okhotsk)

The content of Pb, Cd, Cu, Cr, Mn, Ni, Fe, and As in the muscle, mantle, and digestive glands of the gastropod mollusks Neptunea excelsior, N. behringiana, N. multistriata, and N. varicifera, as well as Buccinum verkruezeni, living near the coasts of the South-Eastern Sakhalin has been estimated. Representatives of the ge-nera Neptunea and Buccinum showed some differences in the ranges of variation and series of value sequences of trace element concentrations in the muscle. Exceeding the hygienic standard for the content of As in the hepatopancreas was found in all the studied mollusks, and in the muscle – in all, except for N. behringiana. The maximum permissible level of Cd concentration was exceeded in the hepatopancreas of N. multistriata.

Integrated lipidomics and transcriptomics analysis reveal lipid metabolism disturbance in scallop (Chlamys farreri) exposure to benzo[a]pyrene

Benzo[a]pyrene (B[a]P) commonly bioaccumulates in lipid-rich tissues due to its lipophilicity and further affects lipid metabolism. The present study systematically investigated the lipid metabolism disturbance in digestive glands of scallops (Chlamys farreri) exposure to B[a]P, based on lipidomics, transcriptomics, molecular and biochemical analysis. We exposed the scallops to environmentally relevant concentrations of B[a]P for 21 days. The bioaccumulation of B[a]P, lipid content and lipid peroxidation in digestive glands were measured. Integrated lipidomics and transcriptomics analysis, the differential lipid species were identified and key genes based on the pathways in which genes and lipid species involved together were selected in scallop exposure to 10 μg/L B[a]P. The changes of lipid profile showed that triglycerides (TGs) were accumulated after 21 days exposure, while the phospholipids (PLs) decreased demonstrated membrane structures were disrupted by B[a]P. In combination with the change of gene expression, we speculated that B[a]P could induce lipids accumulation by up-regulating lipid synthesis-related genes expression, down-regulating lipolysis-related genes expression and interfering with lipid transport. Overall, this study provides new insights into the mechanisms of lipid metabolism disturbance in bivalves exposed to PAHs, and establishes a foundation for understanding the bioaccumulation mechanism of B[a]P in aquatic organisms, which is of great importance for further ecotoxicological study.

Biochemical and histopathological responses of Biomphalaria alexandrina to RIPEX (plant growth regulator)

BackgroundPlant growth regulators are widely used in agriculture for increasing the growth and ripening of plants, but they endanger the aquatic ecosystem. The current study assessed the effect of sublethal exposure to RIPEX 48% EC concentrations (8 and 16 µL/L) on oxidative stress parameters, sex hormones, immune potential enzymes, differential hemocyte counts, and the histopathology of digestive glands and ovotestis in Biomphalaria alexandrina snails.ResultsRIPEX exposure caused an overall increase in Superoxide dismutase and Glutathione-S-Transferase activities in B. alexandrina. However, extreme RIPEX exposure inhibits SOD activity in snails. Malondialdehyde activity showed an increase in B. alexandrina exposed to both concentrations after all exposure periods. RIPEX also caused a significant increase in testosterone in snails exposed to 16 µL/L, it did, however, reduce hormone levels in snails exposed to 8 µL/L at 7 days. Regarding estradiol, there was a significant increase after 3 days of exposure to 16 µl/L and 7 days of exposure to 8 µL/L. RIPEX exposure also increased the activities of Myeloperoxidase and Adenosine deaminase enzymes in the digestive glands of snails. It increased the total hemocyte count of exposed snails as well as the number of granulocytes. Snail digestive glands and ovotestis showed pathological alterations after 7 days of RIPEX exposure.ConclusionsThese findings suggest that RIPEX is toxic to B. alexandrina and that this snail can be used as a bioindicator for environmental contamination with plant growth regulators.

A Genomic and Transcriptomic Analysis of the C-Type Lectin Gene Family Reveals Highly Expanded and Diversified Repertoires in Bivalves

C-type lectins belong to a widely conserved family of lectins characterized in Metazoa. They show important functional diversity and immune implications, mainly as pathogen recognition receptors. In this work, C-type lectin-like proteins (CTLs) of a set of metazoan species were analyzed, revealing an important expansion in bivalve mollusks, which contrasted with the reduced repertoires of other mollusks, such as cephalopods. Orthology relationships demonstrated that these expanded repertoires consisted of CTL subfamilies conserved within Mollusca or Bivalvia and of lineage-specific subfamilies with orthology only between closely related species. Transcriptomic analyses revealed the importance of the bivalve subfamilies in mucosal immunity, as they were mainly expressed in the digestive gland and gills and modulated with specific stimuli. CTL domain-containing proteins that had additional domains (CTLDcps) were also studied, revealing interesting gene families with different conservation degrees of the CTL domain across orthologs from different taxa. Unique bivalve CTLDcps with specific domain architectures were revealed, corresponding to uncharacterized bivalve proteins with putative immune function according to their transcriptomic modulation, which could constitute interesting targets for functional characterization.