Norfloxacin affects inorganic nitrogen compound transformation in tailwater containing Corbicula fluminea.

The Asian clam, Corbicula fluminea, commonly used in engineered wetlands receiving tailwater, affects nitrogen compound transformation in water. This study investigates how a commonly observed antibiotic in tailwater, norfloxacin, impact nitrogen compound transformation in tailwater containing C. fluminea. The clam was exposed to artificial tailwater with norfloxacin (0, 0.2, 20, and 2000 µg/L) for 15 days. Water properties, C. fluminea ecotoxicity responses, microorganism composition and nitrification- or denitrification-related enzyme activities were measured. Results revealed norfloxacin-induced increases and reductions in tailwater NH4+ and NO2- concentrations, respectively, along with antioxidant system inhibition, organ histopathological damage and disruption of water filtering and digestion system. Microorganism composition, especially biodiversity indices, varied with medium (clam organs and exposure water) and norfloxacin concentrations. Norfloxacin reduced NO2- content by lowering the ratio between microbial nitrifying enzyme (decreased hydroxylamine oxidoreductase and nitrite oxidoreductase activity) and denitrifying enzyme (increased nitrate reductase and nitrite reductase activity) in tailwater. Elevated NH4+ content resulted from upregulated ammonification and inhibited nitrification of microorganisms in tailwater, as well as increased ammonia emission from C. fluminea due to organ damage and metabolic disruption of the digestion system. Overall, this study offers insights into using benthic organisms to treat tailwater with antibiotic residues, especially regarding nitrogen treatment.

Comprehensive assessment of multiple biomarker mechanisms in the brackish water clam Corbicula japonica exposed to polystyrene microplastics using structural equation modeling.

Using structural equation modeling (SEM), we investigated multiple biomarker mechanisms in terms of biochemical and individual marker responses in the brackish water clam Corbicula japonica following acute exposure to polystyrene microplastic (PS-MP). This study is the first to comprehensively explore multiple biomarker responses in bivalves using SEM. The model revealed that PS-MP accumulation was an independent biomarker, exhibiting significant direct effects on superoxide dismutase (SOD) and catalase (CAT) among the biochemical markers. Although CAT generally interacts closely with SOD, no significant relationship was identified between them, indicating that CAT may have independently responded to PS-MP stress. Among individual markers, significant indirect effects were observed on clearance rate (CR), reflecting feeding activity and valve open rate, indicating excretion activity via SOD and CAT. Finally, the carbon-based scope for growth was significantly influenced by CR. SEM is efficient and useful for identifying significant direct and indirect pathway relationships and for uncovering uncommon relationships in unified multiple biomarker mechanisms in aquatic studies.

Toxicity of environmental and polystyrene plastic particles on the bivalve Corbicula fluminea: focus on the molecular responses.

Among aquatic organisms, filter feeders are particularly exposed to the ingestion of microplastics (MPs) and nanoplastics (NPs). The present study investigates the effect of environmental microplastics (ENV MPs) and nanoplastics (ENV NPs) generated from macro-sized plastic debris collected in the Garonne River (France), and polystyrene NPs (PS NPs) on the freshwater bivalve Corbicula fluminea. Organisms were exposed to plastic particles at three concentrations: 0.008, 10, and 100 µg L-1 for 21 days. Gene expression measurements were conducted in gills and visceral mass at 7 and 21 days to assess the effects of plastic particles on different functions. Our results revealed: (i) an up-regulation of genes, mainly involved in endocytosis, oxidative stress, immunity, apoptosis, and neurotoxicity, at 7 days of exposure for almost all environmental plastic particles and at 21 days of exposure for PS NPs in the gills, (ii) PS NPs at the three concentrations tested and ENV MPs at 0.008 µg L-1 induced strong down-regulation of genes involved in detoxication, oxidative stress, immunity, apoptosis, and neurotoxicity at 7 days of exposure in the visceral mass whereas ENV MPs at 10 and 100 µg L-1 and all ENV NPs induced less pronounced effects, (iii) overall, PS NPs and ENV MPs 0.008 µg L-1 did not trigger the same effects as ENV MPs 10 and 100 µg L-1 and all ENV NPs, either in the gills or the visceral mass at 7 and 21 days of exposure. This study highlighted the need to use MPs and NPs sampled in the environment for future studies as their properties induce different effects at the molecular level to living organisms.

Harmful planktonic Microcystis and benthic Oscillatoria-induced toxicological effects on the Asian clam (Corbicula fluminea): A survey on histopathology, behavior, oxidative stress, apoptosis and inflammation.

Cyanobacterial blooms are worldwide distributed and threaten aquatic ecosystems and public health. The current studies mainly focus on the adverse impacts of planktonic cyanobacteria or pure cyanotoxins, while the benthic cyanobacteria-induced ecotoxic effects are relatively lacking. The cyanobacterial cell-induced toxic effects on aquatic organisms might be more serious and complex than the pure cyanotoxins and crude extracts of cyanobacteria. This study explored the chronic effects of toxin-producing planktonic Microcystis aeruginosa (producing microcystin) and benthic Oscillatoria sp. (producing cylindrospermopsin) on the behaviors, tissue structures, oxidative stress, apoptosis, and inflammation of the Asian clams (Corbicula fluminea) under 28-d exposure. The data showed that both M. aeruginosa and Oscillatoria sp. can decrease the behaviors associated with the feeding activity and induce tissue damage (i.e. gill and digestive gland) in clams. Furthermore, two kinds of cyanobacteria can alter the antioxidant enzyme activities and increase antioxidant, lipid oxidation product, and neurotransmitter degrading enzyme levels in clams. Moreover, two kinds of cyanobacteria can activate apoptosis-related enzyme activities and enhance the proinflammatory cytokine levels of clams. In addition, two kinds of cyanobacteria can disturb the transcript levels of genes linked with oxidative stress, apoptosis, and inflammation. These results suggested harmful cyanobacteria can threaten the survival and health of clams, while the benthic cyanobacteria-induced adverse effects deserve more attention. Our finding also indicated that it is necessary to focus on the entire algal cell-induced ecotoxicity when concerning the ecological impacts of cyanobacterial blooms.

Ceratophyllum demersum alleviates microplastics uptake and physiological stress responses in aquatic organisms, an overlooked ability.

It has been demonstrated that microplastics (MPs) may be inadvertently ingested by aquatic animals, causing harm to their physiological functions and potentially entering the food chain, thereby posing risks to human food safety. To achieve an environmentally friendly and efficient reduction of MPs in freshwater environments, this experiment investigates the depuration effect of C. demersum on MPs using three common aquatic animals: Macrobrachium nipponense, Corbicula fluminea, and Bellamya aeruginosa as research subjects. The amounts of MPs, digestive enzyme activity, oxidative stress index, and energy metabolism enzyme activity in the digestive and non-digestive systems of three aquatic animals were measured on exposure days 1, 3, and 7 and on depuration days 1 and 3. The results indicated that the depuration effect of C. demersum and the species interaction were significant for the whole individual. Concerning digestive tissue, C. demersum was the most effective in purifying B. aeruginosa. When subjected to short-term exposure to MPs, C. demersum displayed a superior depuration effect. Among non-digestive tissues, C. demersum exhibited the earliest purifying effect on C. fluminea. Additionally, C. demersum alleviated physiological responses caused by MPs. In conclusion, this study underscores C. demersum as a promising new method for removing MPs from aquatic organisms.

Assessing the impact of imidacloprid, glyphosate, and their mixtures on multiple biomarkers in Corbicula largillierti.

Pesticide mixtures are frequently utilized in agriculture, yet their cumulative effects on aquatic organisms remain poorly understood. Aquatic animals can be effective bioindicators and invasive bivalves, owing to their widespread distribution, provide an opportunity to assess these impacts. Glyphosate and imidacloprid, among the most prevalent pesticides globally, are frequently detected in freshwater systems in South America. This study aims to understand the cumulative effects of pesticide mixtures on aquatic organisms, using invasive Corbicula largillierti clams from a natural stream in northwestern Argentina. We conducted 48-hour exposure experiments using two concentrations of imidacloprid (20 and 200 µg L-1 a.i), two concentrations of glyphosate (0.3 and 3 mg L-1 a.i), and two combinations of these pesticides (both at low and high concentrations, respectively), simulating the direct contamination of both pesticides based on their agronomic recipe and observed values in Argentine aquatic environments. Clam metabolism was assessed through the examination of multiple oxidative stress parameters and measuring oxygen consumption rate as a proxy for standard metabolic rate (SMR). Our findings revealed that imidacloprid has a more pronounced effect compared to glyphosate. Imidacloprid significantly decreased clam SMR and cellular levels of reduced glutathione (GSH). However, when both pesticides were present, also cellular glycogen and thiobarbituric acid-reactive substances (TBARS) were affected. Proteins and glutathione S-Transferase (GST) activity were unaffected by either pesticide or their mixture at the assayed concentrations, highlighting the need to test several stress parameters to detect toxicological impacts. Our results indicated additive effects of imidacloprid and glyphosate across all measured parameters. The combination of multiple physiological and cytological biomarkers in invasive bivalves offers significant potential to enhance biomonitoring sensitivity and obtain insights into the origins and cellular mechanisms of chemical impacts. These studies can improve pollution regulatory policies and pesticide management.

Combined effects of toxic Microcystis aeruginosa and high pH on antioxidant responses, immune responses, and apoptosis of the edible freshwater bivalve Corbicula fluminea.

Due to increasing anthropogenic perturbation and water eutrophication, cyanobacterial blooms (CYBs) have become a global ecological and environmental problem. Toxic CYBs and elevated pH are considered to be the two key stressors associated with eutrophication in natural waters, particularly in the event of CO2 depletion induced by dense blooms. However, previous research has been focused on investigating the impacts of toxic CYBs or pH changes in isolation, whereas the interactive effects of such stressors on edible bivalves that inhabit CYB waters still lack information. In this study, the combined effects of toxic Microcystis aeruginosa and pH shifts on the antioxidant responses, immune responses, and apoptosis of the edible freshwater bivalve Corbicula fluminea were explored. The results showed that the activity of antioxidant enzymes was significantly impacted by the interactive effects between toxic M. aeruginosa exposure and time course, yet pH shifts showed no significant effects on the activities of these antioxidant enzymes, implying that the antioxidant response in C. fluminea was mainly triggered by toxic M. aeruginosa exposure. Toxic M. aeruginosa also induced an increased production of reactive oxygen species and malondialdehyde in treated clams, particularly under high pH settings. The elevated lysosomal enzyme activity helped C. fluminea defend against toxic M. aeruginosa exposure under high pH conditions. The principal component analysis (PCA) and the integrated biomarker response (IBR) results suggested that the treated clams were subjected to the elevated toxicity of toxic M. aeruginosa in conditions of high pH. The heat shock proteins-related genes might be triggered to resist the oxidative damage in treated clams. Moreover, the upregulation of TNF and casp8 genes indicated the potential activation of the caspase8-mediated apoptotic pathway through TNF receptor interaction, potentially resulting in apoptosis. The TUNEL assay results further confirmed that apoptosis appeared in treated clams. These findings improve our understanding of the combined toxicological effects of harmful algae and pH shifts on bivalves, which will provide insights into a comprehensive ecological risk assessment of toxic CYBs to edible bivalve species.

Asian clam Corbicula fluminea as potential biomonitor of microplastics and metal(oid)s in a Patagonian River.

This study summarizes the concentration in dry weight (dw) of several metal(oid)s (As, Cd, Cr, Cu, Mn, Ni, Pb, and Zn), as well as the abundance and characteristics of microplastics (MP) in wet weight (ww) of the soft tissues of clam Corbicula fluminea from Chubut River (Patagonia, Argentina). The contents of essential elements were in the following decreasing order: Zn > Mn > Cu > Ni; meanwhile, non-essential elements Cd, Cr and Pb were below the detection limit (<0.5 µg/g dw). A high mean concentration of As (6.1 ± 0.3 µg/g dw) was found, surpassing the maximum allowable limit established by the Argentine Food Code for bivalve molluscs. The number of MP ranged from 0.07 to 1.27 items/ind. and from 0.2 to 2.9 items/g ww. Fibers were the most common shape, mainly transparent. The size of MP ranged from 42 to 1917 µm, accounting for 62 % of MP between 50 and 450 µm. The dominant polymer was PET based on the results of Raman spectroscopy. Based on the widespread distribution of MP in the environment and the wide range of effects on organisms, it is necessary to develop long-term monitoring programs for MP contamination in different environmental matrices. Understanding the bioaccumulation of MP in bivalves is crucial to assess the potential risk to human health through consumption and to the ecosystem. We propose that the widespread Asian clam could serve as a useful biomonitor for MP and As pollution in freshwater and estuarine environments such as the Chubut River.

New insights into the mechanism of ammonia toxicity: Focus on Cactus.

The NF-κB signaling pathway is the most critical pathway in innate immunity. IκB (Cactus) is the primary cytoplasmic inhibitor of NF-κB (Dorsal). In this study, we found that ammonia exposure could significantly induce the expression of Cactus, in a dose-dependent manner in different tissues, with the highest expression in the gill of Corbicula fluminea. The expression pattern-related elements (Tube and Dorsal) in the NF-κB signaling pathway were also analyzed, showing significant up-regulation in 48 h. There was an inhibitory effect between up-regulated Cactus and Dorsal in 72 h, which may regulate Dorsal as a negative feedback pathway function to control the expression of pro-inflammatory cytokines (IL-1ß, IL-6, and TNF-α). Besides, through molecular docking simulation, we found that the Cactus could be directly activated by NH3, complementing the regulatory mechanism of the Cactus. To further test our hypothesis, the levels of pro-inflammatory cytokines decreased after adding PDTC (the antioxidant of Cactus/IκB), suggesting that PDTC can prevent the degradation of Cactus, inhibit Dorsal translocating into the nucleus, and activate the pro-inflammatory cytokines. This revealed the inhibitory effect of Cactus on activating Dorsal/NF-κB factors in the NF-κB signaling pathway. Thus, we suggested that the Cactus is an essential regulator of ammonia-activated inflammation in C. fluminea, which was reported to be activated only by bacteria and immune stimulators. Our study provides a new perspective on the mechanism of ammonia toxicity in invertebrates.

What does the freshwater clam, Corbicula largillierti, have to tell us about chlorothalonil effects?

Chlorothalonil (CLT) is a broad spectrum, and non-systemic fungicide applied in foliar structures to prevent and treat pathogens. This compound reaches to aquatic environments and affects the biota. In this context, the main goal of this study was to assess the effects of CLT at biochemical, tissular, and individual levels of biological organization using the invasive bivalve Corbicula largillierti as a bioindicator species. Clams were exposed to different sublethal concentrations (0, 10, 20 and 50 µg. L-1 CLT) for 96 h. At biochemical level, the enzymatic activity (Glutathione-s-Transferase, Catalase, Acetyl-, Butiryl- and Carboxyl-esterases) and lipid peroxidation were measured in gills and the visceral mass. Also, the digestive gland morphometry through quantitative histological indexes was registered at the tissular level. Finally, filtering activity and burial behavior at the individual level were measured. At the highest CLT concentration, the most significant changes were observed in enzymatic activity (except for butyrylcholinesterase), lipid peroxidation and in digestive gland morphometry. It was also registered increases of the filtering activity and the latency time to burial. Most of the biomarkers assessed showed significant responses under CLT exposure. Therefore, taking into account that C. largillierti was affected by CLT, it can be expected that other species could be in a potential risk if this fungicide is present in freshwater systems.

Characterization of the GABAergic system in Asian clam Corbicula fluminea: Phylogenetic analysis, tissue distribution, and response to the aquatic contaminant carbamazepine.

Gamma-aminobutyric acid (GABA) is the major inhibitory neurotransmitter involved in the neuro-endocrine-immune (NEI) system. In this study, we sequenced the partial length of cDNA fragments of three genes involved in GABA neurotransmitter system of the Asian clam (Corbicula fluminea) (GABAA receptor-associated protein (GABARAP), GABARAPL2 and GABA transporter (GAT-1)). These genes exhibited high amino acid sequence identity compared with other invertebrate orthologs. Expression patterns of the three genes were determined in mantle, gill, gonad, digestive gland and muscle, and the steady state levels of mRNA for each were determined to be highest in gonad and lowest in muscle. To determine their regulation by pharmaceuticals that are present as contaminants in waterways, clams were exposed to carbamazepine (CBZ) for 30 days. CBZ is an agonist for GABA receptors and is an anticonvulsant pharmaceutical that is often detected in aquatic ecosystems. GABARAP and GABARAPL2 mRNA levels were significantly downregulated by 5 and 50 µg/L CBZ in mantle and gill (p < 0.05), while in the gonad and digestive gland, steady state levels (p < 0.05) were decreased with exposure to all three doses. GAT-1 mRNA was upregulated by CBZ (p < 0.05) in the mantle and gill at all three doses tested and in the gonad and digestive system with 5 and 50 µg/L. These data suggest that CBZ disrupt the expression of the GABAergic neurotransmitter system in C. fluminea. Moreover, GABARAP, GABARAPL2 and GAT-1 may be useful biomarkers for the screening of substances that are hazardous to the NEI system of mollusks.

Feeding inhibition following in-situ and laboratory exposure as an indicator of ecotoxic impacts of wildfires in affected waterbodies.

Among the various environmental disturbances caused by wildfires, their impacts within burnt areas and on the downhill aquatic ecosystems has been receiving increased attention. Post-fire rainfalls and subsequent runoffs play an important role in transporting ash and soil to aquatic systems within the burnt areas. These runoffs can be a diffuse source of toxic substances such as metals. The present work aims at assessing the effects of ash-loaded runoff on feeding rates of three representative aquatic invertebrates (Daphnia magna, Corbicula fluminea and Atyaephyra desmarestii) and the mosquitofish, Gambusia holbrooki, through post-exposure feeding inhibition bioassays carried out in-situ and in the laboratory using water collected from the experimental field sites. Four sites were selected in a partially burnt basin for bioassay deployment and sample collection: one site upstream of the burnt area (RUS); three sites receiving runoff directly from the burnt area, one immediately downstream of the burnt area (RDS) and two in permanent tributary streams within the burnt area (BS1 and BS2). The in-situ exposure lasted four days and began following the first post-fire major rain events. At sites affected by the wildfire, post-exposure feeding rates for D. magna, A. desmarestii and G. holbrooki were lower, which is consistent with the highest levels of metals found at these sites compared to the unaffected site, although the individual concentrations of each metal were generally below corresponding ecological safety benchmarks. Thus, interactions between metals and/or between metals and other environmental parameters certainly played a role in modulating the ecotoxic effects of the runoffs; this was further supported by a Toxic Units Summation exercise. Even if direct causal links between the ecotoxicological effects observed in D. magna, A. desmarestii and G. holbrooki and the physicochemical parameters of the water samples could not be established, the results suggest an important role of major and trace elements in explaining post-exposure feeding rate variation.

Proteome changes in muscles, ganglia, and gills in Corbicula fluminea clams exposed to crude oil: Relationship with behavioural disturbances.

The use of online remote control for 24/7 behavioural monitoring can play a key role in estimating the environmental status of aquatic ecosystems. Recording the valve activity of bivalve molluscs is a relevant approach in this context. However, a clear understanding of the underlying disturbances associated with behaviour is a key step. In this work, we studied freshwater Asian clams after exposure to crude oil (measured concentration, 167 ± 28 µg·L-1) for three days in a semi-natural environment using outdoor artificial streams. Three complementary approaches to assess and explore disturbances were used: behaviour by high frequency non-invasive (HFNI) valvometry, tissue contamination with polycyclic aromatic hydrocarbons (PAH), and proteomic analysis. Two tissues were targeted: the pool adductor muscles - retractor pedal muscle - cerebral and visceral ganglia, which is the effector of any valve movement and the gills, which are on the frontline during contamination. The behavioural response was marked by an increase in valve closure-duration, a decrease in valve opening-amplitude and an increase in valve agitation index during opening periods. There was no significant PAH accumulation in the muscle plus nervous ganglia pool, contrary to the situation in the gills, although the latter remained in the low range of data available in literature. Major proteomic changes included (i) a slowdown in metabolic and/or cellular processes in muscles plus ganglia pool associated with minor toxicological effect and (ii) an increase of metabolic and/or cellular processes in gills associated with a greater toxicological effect. The nature of the proteomic changes is discussed in terms of unequal PAH distribution and allows to propose a set of explanatory mechanisms to associate behaviour to underlying physiological changes following oil exposure. First, the first tissues facing contaminated water are the inhalant siphon, the mantle edge and the gills. The routine nervous activity in the visceral ganglia should be modified by nervous information originating from these tissues. Second, the nervous activity in the visceral ganglia could be modified by its own specific contamination. Third, a decrease in nervous activity of the cerebral ganglia close to the mouth, including some kind of narcosis, could contribute to a decrease in visceral ganglia activity via a decrease or blockage of the downward neuromodulation by the cerebro-visceral connective. This whole set of events can explain the decrease of metabolic activity in the adductor muscles, contribute to initiate the catch mechanism and then deeply modify the valve behaviour.

A multibiomarker approach to assess toxic effects of wastewater treatment plant effluents and activated defence mechanisms in marine (Ruditapes philippinarum) and fresh water (Corbicula fluminea) bivalve species.

Since it has been demonstrated that urban effluents can have adverse effects on aquatic organisms, a multibiomarker study was used to evaluate the effects of wastewater treatment plant (WWTP) effluents discharged into the marine and freshwater environments on clams in Cádiz, Spain. One bioassay was performed in the Bay of Cádiz, exposing Ruditapes philippinarum (marine) to a reference site as well as two sites close to WWTP discharges for 14 days. A second bioassay was performed in the Guadalete River, exposing Corbicula fluminea (fresh water) to three sites for 21 days. The biomarkers analysed included defence mechanisms and various toxic effects. Results indicated that WWTP effluents activated defence mechanisms and induced toxic effects in clams exposed to both environments, thus indicating bioavailability of contaminants present in water. Elevated enzymatic activity was found in clams deployed in La Puntilla and El Trocadero compared to control clams and those exposed to the reference site, and 96% of clams deployed at G2 in the Guadalete River died before day 7. Clams exposed to G1 and G3 indicated significant differences in all biomarkers analysed with respect to control clams (p < 0.05). Both species were sensitive to contaminants present in studied sites. This is the first time that these species were used in cages to assess the environmental risk of wastewater effluent discharges in freshwater and marine column environments. The multibiomarker approach provided important ecotoxicological information and is useful for the assessment of the bioavailability and effect of contaminants from WWTP effluents on marine and fresh water invertebrates.

Effects of Bisphenol A on the microalga Chlamydomonas reinhardtii and the clam Corbicula fluminea.

Bisphenol A (BPA) is used throughout the world and it could enter aquatic ecosystems causing harmful effects on humans, animals and plants. The current study relies on the investigation of the toxicity of this emerging pollutant on two freshwater species from different trophic levels: the microalga Chlamydomonas reinhardtii and the clam Corbicula fluminea. After 96 h of exposure to several concentrations of BPA, the growth of C. reinhardtii was affected, being the 96 h-EC50 value for growth 30 mg L-1. The toxicity and bioaccumulation of 30 mg L-1 BPA in microalgae after 24 h of exposure were studied. Several cytotoxicity biomarkers such as vitality, oxidative stress and cytoplasmic membrane potential were altered in exposed cells and microalgae accumulated 0.16 pg BPA cell-1. Regarding C. fluminea, four treatments were established: control without BPA (C); BPA in the food (microalgae pre-exposed for 24 h to 30 mg L-1) (M); BPA in the water (7.5 mg L-1) (W); BPA in both food and water (M + W). After one month of exposure, treated bivalves showed a significantly decrease in the filtration rate and increased lipid peroxidation levels, indicating fitness reduction and oxidative damage. Furthermore, the activities of catalase, glutathione reductase, Se-dependent and total glutathione peroxidase enzymes increased significantly in W and M + W treatments with respect to the control. Clams of the M + W treatment were the most affected, indicating that the little amount of BPA bioaccumulated by microalgae could increase the damage. Emerging contaminants may accumulate in several organisms, such as microalgae, and could have negative impacts on ecosystems.

Effects of acute ammonia toxicity on oxidative stress, DNA damage and apoptosis in digestive gland and gill of Asian clam (Corbicula fluminea).

Ammonia is one of the major pollutants associated with the main river basins due to ammonification of uneaten food and animal excretion, which usually brings detrimental health effects to aquatic invertebrate. However, the mechanisms of ammonia toxicity in aquatic invertebrate have rarely been reported. In this study, C. fluminea was exposed to different levels of ammonia (control group, 10 mg/L, and 25 mg/L) for 24 h and 48 h, and digestive gland and gill were collected to explore toxic effects on oxidative stress, DNA damage and apoptosis under ammonia stress. The results showed that ammonia poisoning could increase the activity of oxidative stress enzyme (SOD and CAT), inducing differentially expressed genes (DRAM2, GADD45, P53, BAX, BCL2, CASP8, CASP9, CASP3, HSP70 and HSP90) and different cytokines (IL-1 beta, IL-8, IL-17 and TNF-alpha) of DNA damage and apoptosis. The difference of toxic effects induced by ammonia among digestive gland and gill were also observed by real-time PCR and TUNEL staining. Our results will be helpful to understand the mechanism of aquatic toxicology induced by ammonia in C. fluminea.

Effect of imidacloprid on the behavior, antioxidant system, multixenobiotic resistance, and histopathology of Asian freshwater clams (Corbicula fluminea).

In the current study, to investigate the effect of imidacloprid on benthic bivalves, adult Asian clams (Corbicula fluminea) were exposed to 0, 20, 200, and 2000 µg/L imidacloprid for 30 d. Imidacloprid significantly inhibited the siphoning and burrowing behaviour (p < 0.05) of the clams. Significant histopathological changes were associated with degeneration of the cilium, the contraction and adhesion of the lymphocyte, and the swelling of epithelium cells in gills, and there was notable degeneration in the digestive tubules, haemolytic infiltration in the connective tissue and epithelial cell necrosis in the digestive glands in the 2000 µg/L treatment group. The activity of AChE in the digestive glands was significantly inhibited at all treatment levels, whereas this inhibition was observed in gills only in the 2000 µg/L treatment (p < 0.05). Additionally, indicators of the antioxidant system (e.g., SOD, CAT, and GST activity) and MDA content were significantly increased in the gills and digestive glands with all treatments (p < 0.05). Moreover, the mRNA expression levels of Hsp genes (hsp 22, hsp 40, hsp 60, hsp 70, hsp 90) and multixenobiotic resistance (MXR) system-related genes (abcb1, abcc1) were significantly downregulated (p < 0.05). Therefore, our results suggest that imidacloprid changes the oxidative stress, cellular detoxification, and MXR system of C. fluminea. Our findings provide new insights into the effects of neonicotinoids on benthic bivalves such as C. fluminea.

Asiatic clam Corbicula fluminea exhibits distinguishable behavioural responses to crude oil under semi-natural multiple stress conditions.

Aquatic ecosystems are subject to many anthropogenic disturbances, and understanding their possible impacts is a real challenge. Developing approaches based on the behaviour of bivalve mollusks, an integrating marker of the state of the organisms, and therefore of their environment, is relevant, whether within a natural ecosystem or an ecosystem subject to industrial activities. The main objective of this study was to identify by HFNI Valvometry a reliable and reproducible clam behavioural response in the presence of crude oil in a multistress context. To closely replicate actual field conditions, Corbicula fluminea was exposed in outdoor artificial streams that were subject to natural variations and were continuously fed by fresh water from the Gave de Pau (S.W. France). After a period of 26 days in these artificial streams, the clams (n = 14-16 per condition) were separately exposed for 10 days to crude oil alone, crude oil and barium, crude oil and noise pollution, crude oil and turbidity pulses, barium alone, noise pollution alone, turbidity pulses alone or natural changes alone. The secondary objective was to characterize the accumulation of polycyclic aromatic hydrocarbons (PAH) in 3 tissues (gills, adductor muscles and foot) in clams exposed for 10 days to crude oil alone or under multistress conditions (n = 5 clams per condition) and then to compare the accumulation and behaviour of clams under these conditions. The response of clams to crude oil alone or under multistress conditions was visually and statistically significant and not confounded by the other disturbances tested, despite large variations in water temperature. In the presence of crude oil, the behaviour of clams was characterized by an increase in valve-closure duration, a decrease in valve-opening amplitude and an increase in valve agitation index. In the presence of crude oil, the clam behaviour showed no direct relationship with PAH accumulation in the gills, adductor muscles or foot, although hypothetical mechanisms are discussed. This work supports the growing interest in studying the behaviour of bivalve mollusks in the context of biomonitoring of the aquatic environment surrounding oil facilities.

An Assessment of Heavy Metals Toxicity in Asian Clam, Corbicula fluminea, from Mekong River, Pa Sak River, and Lopburi River, Thailand.

High population density and economic development attributing to the changes in water quality in Pa Sak River, Lopburi River, and Mekong River have attracted great attention. This research aimed to determine the pollution of heavy metals in collected clams at three different study sites. Bioaccumulation of heavy metals in Asian clam (Corbicula fluminea) may be likely to cause serious health effects on human beings. The clams sampled from three different rivers (Mekong, Pa Sak, and Lopburi) from Thailand were analyzed for the presence of heavy metals (Zn, Cu, Cd, Cr, Mn, and Pb) with an air-acetylene flame atomic absorption spectrophotometer (AAS). Among the heavy metals studied, Zn was recorded as having the highest concentration (127.33-163.65 µg/g) among the three rivers. The observed mean concentration of Cu was in the range of 84.61-127.15 µg/g followed by Mn (13.96-100.63 µg/g), Cr (5.79-15.00 µg/g), Pb (3.43-8.55 µg/g), and Cd (0.88-1.95 µg/g). Overall, Asian clam from Pa Sak River was found to contain high concentrations of Zn, Cu, Cd, Cr, and Pb compared to Mekong and Lopburi River.

Transcriptome analysis of response mechanism to ammonia stress in Asian clam (Corbicula fluminea).

Corbicula fluminea is highly sensitive to ammonia, and its response mechanism to ammonia stress is unclear. In this study, C. fluminea was exposed to different levels of ammonia (control group, 10 mg/L, and 25 mg/L) for 24 h and 48 h. A comparative analysis of transcriptome sequencing (RNA-seq) of C. fluminea digestive gland showed that the expression of 6742 genes (11.54%) was significantly affected by ammonia stress. The TLR, NF-κB, FOXO, and apoptotic signaling pathways were involved in the regulation. The differential expression of 14 genes was confirmed by real-time PCR. In summary, the response mechanism of C. fluminea digestive gland under ammonia stress may be different from that of oxidative stress in marine vertebrates. Also, the NMDAR-mediated pathway may not be the main mechanism in the response to ammonia stress in C. fluminea. The present study is a preliminary study for further investigation into ammonia toxicity in shellfish.