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.

Spectroscopic studies for identifying the chemical states of the periostracum of the Corbicula species in Lake Biwa.

Corbicula clam shells consist of thin periostracum and calcareous layers made of calcium carbonate (CaCO3). Depending on habitat conditions, the shell exhibits various colorations, such as yellow, brown, and black. The chemical state of the periostracum of the Corbicula species in Lake Biwa was studied by X-ray absorption fine structure (XAFS) and Raman scattering spectroscopies. Fe K-edge X-ray absorption near edge structure (XANES) revealed that the Fe3+ intensity increases as the color of the shell changes from yellow to black. Raman spectra suggested that quinone-based polymers cover the yellow shell, and the black shell is further covered by dihydroxyphenylalanine (DOPA) rings of amino acid derivatives. From Fe K-edge extended X-ray absorption fine structure (EXAFS), it was found that Fe3+ in the periostracum was surrounded by five to six oxygen atoms with an average Fe-O ligand distance of 2.0 Å. Accordingly, a tris-DOPA-Fe3+ complex is formed, which is responsible for the periostracum's black color.

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.

Exposure to Oxidized Multi-Walled CNTs Can Lead to Oxidative Stress in the Asian Freshwater Clam Corbicula fluminea (Müller, 1774).

The increasing attention that carbon-based nanomaterials have attracted due to their distinctive properties makes them one of the most widely used nanomaterials for industrial purposes. However, their toxicity and environmental effects must be carefully studied, particularly regarding aquatic biota. The implications of these carbon-based nanomaterials on aquatic ecosystems, due to their potential entry or accidental release during manufacturing and treatment processes, need to be studied because their impacts upon living organisms are not fully understood. In this research work, the toxicity of oxidized multi-walled carbon nanotubes (Ox-MWCNTs) was measured using the freshwater bivalve (Corbicula fluminea) after exposure to different concentrations (0, 0.1, 0.2, and 0.5 mg·L-1 Ox-MWCNTs) for 14 days. The oxidized multi-walled carbon nanotubes were analyzed (pH, Raman microscopy, high-resolution electron microscopy, and dynamic light scattering), showing their properties and behavior (size, aggregation state, and structure) in water media. The antioxidant defenses in the organism's digestive gland and gills were evaluated through measuring oxidative stress enzymes (glutathione-S-transferase, catalase, and superoxide dismutase), lipid peroxidation, and total ubiquitin. The results showed a concentration-dependent response of antioxidant enzymes (CAT and GST) in both tissues (gills and digestive glands) for all exposure periods in bivalves exposed to the different concentrations of oxidized multi-walled carbon nanotubes. Lipid peroxidation (MDA content) showed a variable response with the increase in oxidized multi-walled carbon nanotubes in the gills after 7 and 14 exposure days. Overall, after 14 days, there was an increase in total Ub compared to controls. Overall, the oxidative stress observed after the exposure of Corbicula fluminea to oxidized multi-walled carbon nanotubes indicates that the discharge of these nanomaterials into aquatic ecosystems can affect the biota as well as potentially accumulate in the trophic chain, and may even put human health at risk if they ingest contaminated animals.

Intrahabitat Differences in Bacterial Communities Associated with Corbicula fluminea in the Large Shallow Eutrophic Lake Taihu.

The Asian clam Corbicula fluminea is a keystone zoobenthos in freshwater ecosystems. However, its associated microbiome is not well understood. We investigated the bacterial communities of this clam and its surrounding environment, including sediment and water simultaneously, in a large lake by means of 16S rRNA gene sequencing. Approximately two-thirds of the bacterial operational taxonomic units (OTUs) associated with clams were observed in the surrounding environment and mostly from particle-associated samples. The associated bacterial communities were site specific and more similar to environmental bacteria from the same site than those at other sites, suggesting a local environmental influence on host bacteria. However, the significant differences in bacterial diversities and compositions between the clam and the environment also indicated strong host selection pressure on bacteria from the surrounding environment. Bacteria affiliated with Firmicutes, Spirochaetes, Tenericutes, Bacteroidetes, Epsilonbacteraeota, Patescibacteria, and Fusobacteria were found to be significantly enriched in the clams in comparison to their local environment. Oligotyping analyses of the core-associated bacterial OTUs also demonstrated that most of the core OTUs had lower relative abundances and occurrence frequencies in environmental samples. The core bacterial OTUs were found to play an important role in maintaining the stability of the bacterial community network. These core bacteria included the two most abundant taxa Romboutsia and Paraclostridium with the potential function of fermenting polysaccharides for assisting host clams in food digestion. Overall, we demonstrate that clam-associated bacteria were spatially dynamic and site specific, which were mainly structured both by local environments and host selection. IMPORTANCE The Asian clam Corbicula fluminea is an important benthic clam in freshwater ecosystems due to its high population densities and high filtering efficiency for particulate organic matter. While the associated microbiota is believed to be vital for host living, our knowledge about the compositions, sources, and potential functions is still lacking. We found that C. fluminea offers a unique ecological niche for specific lake bacteria. We also observed high intrahabitat variation in the associated bacterial communities. Such variations were driven mainly by local environments, followed by host selection pressure. While the local microbes served as a source of the clam-associated bacteria, host selection resulted in enrichments of bacterial taxa with the potential for assisting the host in organic matter digestion. These results significantly advance our current understanding of the origins and ecological roles of the microbiota associated with a keynote clam in freshwater ecosystems.

Seasonal trends of mercury bioaccumulation and assessment of toxic effects in Asian clams and microbial community from field study of estuarine sediment.

This study investigated seasonal trends in bioaccumulation potential and toxic effects of mercury (Hg) in Asian clams (Corbicula fluminea) and microbial community. For this, a clam-exposure experiment was performed during summer, fall, and winter seasons in four different sites (HS1: control/clean site; HS2, HS3, and HS4: contaminated sites) of Hyeongsan River estuary, South Korea. Total mercury (THg) and methylmercury (MeHg) in whole sediments were highest at HS4 site during fall, sustained similar levels during winter, but decreased during summer. Unlike whole sediment, pore water reported higher levels in summer, and gradually declined during fall and winter. Asian clams from HS4 site collected during summer presented highest bioaccumulations of THg (521.52 µg/kg, dry weight) and MeHg (161.04 µg/kg, dry weight), which also correlated with the higher levels of Hg present in pore water in the same season. Moreover, biota-sediment-pore water accumulation factor (BSpAF) were comparatively greater in clams collected from HS2∼HS4 compared to HS1 sites, suggesting that porewater was a better indicator of accumulation of Hg. Upregulation of biomarker genes responsible for detoxifying process (gsts1), scavenging oxidative stress (cat), and protein reparation (hsp70 and hsp90) were observed in clams collected from HS2∼HS4. The overexpression of these biomarkers implied that Asian clams can be considered as promising warning tools for Hg-contamination. Both bacterial and metabolic diversities were negatively affected by higher levels of THg and MeHg. Phylum Proteobacteria was enriched in HS2∼HS4 compared to HS1. In contrast, phylum Bacteroidetes showed a reverse trend. The metabolic profile was highest in HS1 and lowest in HS4, revealing higher stress of Hg in HS4 site. Overall, the outcomes of this field study broaden the information on seasonal trends of bioaccumulation of Hg and its toxic effects. These findings may be helpful in Hg monitoring and management programs in other river systems.

The Effects of Freshwater Clam (Corbicula fluminea) Extract on Serum Tumor Necrosis Factor-Alpha (TNF-α) in Prediabetic Patients in Taiwan.

Freshwater clam extract (FCE) is a functional food that regulates the immune system and has been demonstrated in numerous studies to display desirable anti-tumor necrosis factor-alpha (TNF-α) responses. In addition, excess TNF-α production is positively associated with type 2 diabetes. However, few longitudinal clinical studies evaluating the efficiency and toxicity of FCE are available. This article reports that patients with prediabetes who received FCE had a desirable outcome of a reduction in serum TNF-α for a long period. This was a double-blind, randomized, parallel clinical trial conducted using FCE intervention and placebo groups, and 36 patients with prediabetes were enrolled. Two grams of FCE or placebo was consumed daily for 180 consecutive days. The serum of the participants was collected at four time points (0M: before the intervention; 3M: after 3 months of intervention; 6M: after 6 months of intervention; 12M: 6 months after cessation of intervention at 6M). A serum TNF-α concentration higher than 4.05 pg/mL was defined as a cut-off value. FCE reduced serum TNF-α in all participants at 6M and 12M. Moreover, FCE significantly suppressed serum TNF-α concentrations at 6M and 12M and inhibited TNF-α release with time series in subjects with elevated TNF-α values. FCE intervention effectively reduced serum TNF-α and persistently sustained the effects for half a year in patients with prediabetes. Gas chromatography-mass spectrometry (GS-MS) analysis revealed that the major components of FCE were phytosterols and fatty acids, which exerted anti-inflammatory and anti-TNF-α abilities. Hence, FCE has the potential to be developed as a natural treatment for prediabetic patients in Taiwan.

PacBio Full-Length and Illumina Transcriptomes of the Gill Reveal the Molecular Response of Corbicula fluminea under Aerial Exposure.

Air exposure is a common stress for Corbicula fluminea, an economically important freshwater shellfish consumed in China, during aquaculture and transportation. However, little is known about its molecular responses to air exposure. Therefore, this study used a combination of PacBio full-length and Illumina transcriptomes to investigate its molecular responses to air exposure. A total of 36,772 transcripts were obtained using PacBio sequencing. Structural analysis identified 32,069 coding sequences, 1906 transcription factors, 8873 simple sequence repeats, and 17,815 long non-coding RNAs. Subcellular localization analysis showed that most transcripts were located in the cytoplasm and nucleus. After 96-h of air exposure, 210 differentially expressed genes (DEGs) in the gill were obtained via Illumina sequencing. Among these DEGs, most of the genes related to glycolysis, tricarboxylic acid cycle, lipid metabolism, and amino acid metabolism were upregulated. Additionally, many DEGs associated with immunity, cytoskeleton reorganization, autophagy, and ferroptosis were identified. These findings indicated that metabolic strategy change, immune response, cytoskeleton reconstruction, autophagy, and ferroptosis might be the important mechanisms that C. fluminea use to cope with air exposure. This study will enrich the gene resources of C. fluminea and provide valuable data for studying the molecular mechanisms coping with air exposure in C. fluminea and other freshwater mollusks.

Hypoglycemic peptide-enriched hydrolysates of Corbicula fluminea and Chlorella sorokiniana possess synergistic hypoglycemic activity through inhibiting α-glucosidase and dipeptidyl peptidase-4 activity.

BACKGROUND: The prevalence of diabetes mellitus worldwide has increased in recent decades. Maintaining the level of blood glucose is the most basic and important issue for diabetics. This study aimed to investigate the hypoglycemic activity of a combination of hypoglycemic peptide-enriched hydrolysates of Corbicula fluminea (ACH) and Chlorella sorokiniana (PCH). RESULTS: Combined supplementation of ACH and PCH synergistically inhibited α-glucosidase and DPP4 activities in vitro. After 4 weeks of treatment with ACH and/or PCH, the plasma glucose concentration and insulin, homeostasis model assessment-estimated insulin resistance (HOMA-IR), total cholesterol (TC) and triglyceride (TG) levels significantly decreased. The hypoglycemic peptides in ACH and PCH were purified and assayed for α-glucosidase and DPP4 activity. The hypoglycemic peptides in ACH and PCH effectively decreased α-glucosidase and DPP4 activities. In silico assays showed that these two peptide types have different docking poses, which determined their inhibitory effect against α-glucosidase and DPP4 activity. CONCLUSION: Combined treatment with hypoglycemic peptide-enriched ACH and PCH could modulate blood glucose by synergistically inhibiting α-glucosidase and DPP4 activities. © 2021 Society of Chemical Industry.

Application of a Multi-Metal Stable-Isotope-Enriched Bioassay to Assess Changes to Metal Bioavailability in Suspended Sediments.

The direct measurement of particulate contaminant bioavailability is a challenging aspect for the environmental risk assessment of contaminated sites. Here, we demonstrated a multi-metal stable-isotope-enriched bioassay to simultaneously measure the bioavailability of Cd, Cu, and Zn in naturally contaminated sediments following differing periods of resuspension treatment. Freshwater filter-feeding clams were pre-labeled with the isotopes 114Cd, 65Cu, and 68Zn to elevate isotope abundances in their tissues and then exposed to metal-contaminated suspended sediments. The assimilation of sediment-associated metals by clams would decrease the isotope ratios (Cd114/111, Cu65/63, and Zn68/64) in tissues, providing a direct measurement of metal bioavailability. For the sediments tested here, the method revealed bioavailable cadmium and non-bioavailable copper in sediments but was inconclusive for zinc. With a longer resuspension time, the bioavailability of particulate cadmium increased, but that of copper was unaffected. Metal bioavailability predicted using traditional wet-chemical extraction methods was inconsistent with these findings. The study indicated that multi-metal stable-isotope-enriched bioassay provides a new tool for directly assessing metal bioavailability in sediments, and this method is amenable for use in in situ assessments.

Acorbine, a Corbicula japonica-derived tripeptide containing non-proteinogenic amino acids, suppresses ethanol-induced liver injury.

Since ancient times, Corbicula extract has been believed in Japan to have hepatoprotective effects, but it remains unclear whether these claims are true, and if so, which component is responsible for hepatoprotection. In this study, we showed that Corbicula extract exerted a protective effect against liver damage. Recent work identified acorbine (ß-alanyl-ornithyl-ornithine), a novel tripeptide containing non-proteinogenic amino acids, in the extract of Corbicula japonica. Synthesized acorbine cured alcohol-induced liver damage in mice. In addition, acorbine purified from Corbicula extract exerted a protective effect against alcohol-induced hepatotoxicity in a culture liver model derived from mouse ES/iPS cells. Thus, acorbine is one of the components of Corbicula extract that protects hepatocytes against ethanol-induced death.

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.

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 microplastics and mercury in the freshwater bivalve Corbicula fluminea (Müller, 1774): Filtration rate, biochemical biomarkers and mercury bioconcentration.

The main objectives of this study were to investigate the effects of a mixture of microplastics and mercury on Corbicula fluminea, the post-exposure recovery, and the potential of microplastics to influence the bioconcentration of mercury by this species. Bivalves were collected in the field and acclimated to laboratory conditions for 14 days. Then, a 14-day bioassay was carried out. Bivalves were exposed for 8 days to clean medium (control), microplastics (0.13 mg/L), mercury (30 µg/L) and to a mixture (same concentrations) of both substances. The post-exposure recovery was investigated through 6 additional days in clean medium. After 8 and 14 days, the following endpoints were analysed: the post-exposure filtration rate (FR); the activity of cholinesterase enzymes (ChE), NADP-dependent isocitrate dehydrogenase (IDH), octopine dehydrogenase, catalase, glutathione reductase, glutathione peroxidase and glutathione S-transferases (GST), and the levels of lipid peroxidation (LPO). After 8 days of exposure to mercury, the bioconcentration factors (BCF) were 55 in bivalves exposed to the metal alone and 25 in bivalves exposed to the mixture. Thus, microplastics reduced the bioconcentration of mercury by C. fluminea. Bivalves exposed to microplastics, mercury or to the mixture had significantly (p ≤ 0.05) decreased FR and increased LPO levels, indicating fitness reduction and lipid oxidative damage. In addition, bivalves exposed to microplastics alone had significant (p ≤ 0.05) reduction of adductor muscle ChE activity, indicating neurotoxicity. Moreover, bivalves exposed to mercury alone had significantly (p ≤ 0.05) inhibited IDH activity, suggesting alterations in cellular energy production. Antagonism between microplastics and mercury in FR, ChE activity, GST activity and LPO levels was found. Six days of post-exposure recovery in clean medium was not enough to totally reverse the toxic effects induced by the substances nor to eliminate completely the mercury from the bivalve's body. These findings have implications to animal, ecosystem and human health.

Whole-transcriptome response to wastewater treatment plant and stormwater effluents in the Asian clam, Corbicula fluminea.

The increase in human population and urbanization are resulting in an increase in the volume of wastewater and urban runoff effluents entering natural ecosystems. These effluents may contain multiple pollutants to which the biological response of aquatic organisms is still poorly understood mainly due to mixture toxicity and interactions with other environmental factors. In this context, RNA sequencing was used to assess the impact of a chronic exposure to wastewater treatment plant and stormwater effluents at the whole-transcriptome level and evaluate the potential physiological outcomes in the Asian clam Corbicula fluminea. We de-novo assembled a transcriptome from C. fluminea digestive gland and identified a set of 3,181 transcripts with altered abundance in response to water quality. The largest differences in transcriptomic profiles were observed between C. fluminea from the reference site and those exposed to wastewater treatment plant effluents. On both anthropogenically impacted sites, most differentially expressed transcripts were involved in signaling pathways in relation to energy metabolism such as mTOR and FoxO, suggesting an energy/nutrient deficit and hypoxic conditions. These conditions were likely responsible for damages to proteins and transcripts in response to wastewater treatment effluents whereas exposure to urban runoff might result in immune and endocrine disruptions. In absence of comprehensive chemical characterization, the RNAseq approach could provide information regarding the mode of action of pollutants and then be useful for the identification of which parameters must be studied at higher integration level in order to diagnose sites where the presence of complex and variable mixtures of chemicals is suspected.

Integrative assessment of sediment quality in lower basin affected by former mining in Brazil.

The Ribeira de Iguape River (Southeast Brazil) is metal contaminated by mining activities. Despite it has been cataloged as "in via of restoration" by the literature, this basin is still a sink of pollution in some segments of the fluvial system. This study aimed to assess the sediment quality in the lower part of the RIR basin. The employed approach was based on biological responses of the freshwater clam Corbicula fluminea after 7-day exposure bioassays using as the reference site the Perequê Ecological Park. Toxic responses (burial activity and lethality) and biochemical biomarkers (GST, GR, GPx, LPO, MTs, AChE and DNA damage) were evaluated and then integrated with metal bioavailability and chemical concentrations to address the sediment quality in the area through the weight-of-evidence approach. A multivariate analysis identified linkages between biological responses and contamination. Results pointed that, despite being below the benchmarks of the US Environmental Protection Agency, there is slight metal contamination in the lower part of the basin which induces oxidative stress in C. fluminea; other toxic responses were sometimes attributed to As and Cr bioaccumulation. The sediment quality values (TEL-PEL values in mg/kg) were calculated for the current study for As (0.63-1.31), Cr (3.5-11.05), Cs (1.0-1.17), Cu (6.32-7.32), Ni (6.78-7.46), Ti (42.0-215), V (1.77-8.00). By comparison with other international guidelines, the sediment quality of the lower basin of the Vale de Ribeira does not identify a significant environmental risk.

Freshwater clam extract supplementation improves wound healing by decreasing the tumor necrosis factor α level in blood.

BACKGROUND: The freshwater clam (Corbicula fluminea) is a widely consumed functional food in Asia and is traditionally used to improve health and either prevent or treat inflammation-related diseases. Numerous studies have proposed that freshwater clams act to prevent and attenuate inflammatory responses, and also serve as a possible inhibitor to systemic inflammation. However, there is limited information available about the effects of freshwater clams on wound healing. RESULTS: The present study investigated the influence of freshwater clam extract (FCE) on wound healing and inflammatory responses in a cutaneous incision model. Sixteen rats were used and divided into two groups: the FCE group and the normal saline (NS) group. The rats underwent dorsal full-thickness skin excisional wounds (diameter 20 × 10 mm). FCE or NS was administered for oral feeding twice daily for 14 days after wounding. Blood samples were taken and analyzed, and wound areas were measured at several time points during the 2 weeks after excision. On day 14 after wounding, skin biopsies from the wound sites were sent for histological examination. Treatment with FCE (71.63 ± 9.51 pg mL-1 ) decreased tumor necrosis factor-α levels compared to the NS group (109.86 ± 12.55 pg mL-1 ) after wounding at 3 h (P < 0.05). There were no significant differences in the levels of white blood cells, interleukin (IL)-6, or IL-10. The wound areas of the NS group (23.9%) were larger than those in the FCE group (8.26%) on day 14 (P < 0.05). Numerous fibroblasts and collagen fiber organization were observed in the FCE group. CONCLUSION: FCE supplementation improves the wound healing process. © 2016 Society of Chemical Industry.

In vitro test systems supporting the development of improved pest control methods: a case study with chemical mixtures and bivalve biofoulers.

Using the bivalve macrofouler Corbicula fluminea, the suitability of in vitro testing as a stepping stone towards the improvement of control methods based on chemical mixtures was addressed in this study. In vitro cholinesterase (ChE) activity inhibition following single exposure of C. fluminea tissue to four model chemicals (the organophosphates dimethoate and dichlorvos, copper and sodium dodecyl phosphate [SDS]) was first assessed. Consequently, mixtures of dimethoate with copper and dichlorvos with SDS were tested and modelled; mixtures with ChE revealed synergistic interactions for both chemical pairs. These synergic combinations were subsequently validated in vivo and the increased control potential of these selected combinations was verified, with gains of up to 50% in C. fluminea mortality relative to corresponding single chemical treatments. Such consistency supports the suitability of using time- and cost-effective surrogate testing platforms to assist the development of biofouling control strategies incorporating mixtures.

The use of a Weight-of-Evidence approach to address sediment quality in the Odiel River basin (SW, Spain).

The fluvial systems of the Iberian Pyrite Belt (SW Iberian Peninsula) are affected by acid mine drainage (a lixiviate residue product of mining activities derived from sulfide oxidation). The high acidity and high concentrations of sulfates and metal(loid)s are the main causes of the environmental degradation of the Odiel River basin. The use of weight of evidence approach in areas of the Odiel River basin implies the integration of different lines of evidence (chemistry, toxicity and bioaccumulation) using the freshwater clam Corbicula fluminea as target species. The integration of the results from the application of the different lines of evidence showed that the index of pollution was higher downvalley (Ptriad=12,312), moderate after mining effluent discharges (Ptriad=13.9) and very low where the Odiel River sources (Ptriad=6.31). The multivariate analysis indicated that variables and chemicals were associated with geochemical matrix and background levels (% of fines and toxic metal(loid) concentrations), toxic effects, and metal(loid) bioaccumulation reflecting the geographical distribution of the contamination towards the estuary. Metal(loid) thresholds were calculated for the study area as site-specific values of interim freshwater sediment quality values: As≥171; Cd≥0.48; Co≥8.82; Cr≥38.4; Cu≥451; Ni≥18.4; Pb≥377; Sb≥17.7; Zn≥221mg/kg of freshwater fluvial dry sediment. These results revealed the possibility of using the TEL values proposed by the USEPA and the NOAA for sediments from this site, and proposed lower PEL values for the Iberian Pyrite Belt as result of toxicity effects found in the Asian clam due to the combination of extreme acidity and high metal(loid) concentrations.

Assessing the potential of the European Atlantic sturgeon Acipenser sturio to control bivalve invasions in Europe.

This pilot study explored the potential of juvenile European Atlantic sturgeon Acipenser sturio to feed on two invasive bivalve species, the Asian clam Corbicula fluminea and the Eurasian zebra mussel Dreissena polymorpha. Preliminary results indicate that native A. sturio were feeding on D. polymorpha at a very limited rate and their potential to prevent the establishment of invasive bivalve species, in new and previously invaded areas, is considered limited.