NO production of granulocytes associated with antibacterial immune response in Tegillarca granosa.

Nitric oxide (NO) is a signaling molecule and immune effector produced by the nitric oxide synthases (NOS), which involved to various physiological processes of animals. In marine bivalves, hemocytes play important roles in antimicrobial innate immune response. Although hemocyte-derived NO has been detected in several bivalves, the immune function of hemocyte-derived NO is not well understood. Here, we investigated the antibacterial response of hemocyte-derived NO in the blood clam Tegillarca granosa. Two types of hemocytes including erythrocytes and granulocytes were isolated by Percoll density gradient centrifugation, their NO production and TgNOS expression level were analyzed. The results showed that NO was mainly produced in granulocytes and almost no detected in erythrocytes. The granulocytes showed significantly higher NO level and TgNOS expression level than the erythrocytes. And the TgNOS expression level was significantly increased in granulocytes after Vibro parahemolyticus challenge. In addition, the NO donor sodium nitroprusside (SNP) significantly increased the NO production of hemocytes to kill pathogenic bacteria. In summary, the results revealed that granulocytes-derived NO play vital roles in the antimicrobial immune response of the blood clam.

Two common nanoparticles exert immunostimulatory and protective effects in Tegillarca granosa against Vibrio parahaemolyticus.

There are many studies revealed that metal-based nanoparticles (NPs) possess excellent bactericidal effect on multitudinous bacteria and fungi. However, the control effect of NPs as antimicrobial agents to against Vibrio parahaemolyticus infection remain in poorly understood for blood clam, Tegillarca granosa. In order to evaluate the effect, the changes in six physiological parameters and the immune-related genes expression of clams exposed to V. parahaemolyticus alone or along with NPs (nZnO or nCuO) were investigated in present study. Results showed that both tested NPs exerted prominent redemptive or mitigative effect in an inverse dose-dependent way on physiological indexes of clam, especially in the total counts, phagocytosis and the cell viability of haemocytes, as well as the concentration and activity of lysozymes, when co-exposed with Vibrio. Gene expression analysis showed NPs at a concentration of 0.1 mg/L generally mitigated the downregulation of immune-related genes after clam exposure to V. parahaemolyticus. The combination of 0.1 mg/mL nZnO and nCuO additives has been shown to significantly enhance the humoral immunity of blood clam, suggesting its potential as a protective measure against V. parahaemolyticus infection in T. granosa.

Chemical and electrochemical conversion of magnetic nanoparticles to Prussian blue for label-free and refreshment-enhanced electrochemical biosensing of enrofloxacin.

Magnetic biosensor takes advantage of rapid and facile magnetic separation/collection of targets, however, generally relies on additional signal labels to generate signal in a tedious and high-cost way. Here, we proposed a chemical and electrochemical conversion (C-ECC) method to develop a label-free electrochemical magnetic biosensor to detect antibiotics enrofloxacin (ENR). The C-ECC method integrates the chemical decomposition of magnetic beads (MBs) to release ironic ions and the simultaneous electrochemical deposition of Prussian blue (PB) analogs through the reaction of ironic ions and co-existing K4Fe(CN)6. Unlike conventional method that relies on the physical magnetic property of MBs, the C-ECC method fully exploited the chemical/electrochemical properties of MBs to produce electrochemically active PB to generate signal, thus endowing MBs with dual roles in both sample treatment and signal generation. The incorporation of chemical and electrochemical conversion produced more PB with higher electroactivity when compared with sole chemical or electrochemical conversion. Moreover, an interesting electrochemical refreshment (ER) was designed to remove insulative species on the electrode surface to improve electroactivity of electrode and benefit amperometric detection significantly. Under optimized conditions, the C-ECC-based biosensor presented limit of detection (LOD) of 4.17 pg mL-1 for ENR, which is lower than most analogs, as well as satisfactory specificity. The biosensor also performed well in fish and chicken meat samples, with LODs lower than maximum residue limits of national standards. The C-ECC method may create a new way to design magnetic sensors and contribute to rapid, facile and sensitive detection in agriculture/food, clinic diagnosis and environmental monitoring.

Integrated RNA-seq and RNAi Analysis of the Roles of the Hsp70 and SP Genes in Red-Shell Meretrix meretrix Tolerance to the Pathogen Vibrio parahaemolyticus.

The "Wanlihong" Meretrix meretrix (WLH-M) clam is a new variety of this species that has a red shell and stronger Vibrio tolerance than ordinary M. meretrix (ORI-M). To investigate the molecular mechanisms responsible for the WLH-M strain's tolerance to Vibrio, we challenged clams with Vibrio parahaemolyticus and then assessed physiological indexes and conducted transcriptome analysis and RNA interference experiments. The mortality, tissue bacterial load, and hemocyte reactive oxygen species level of ORI-M were significantly higher than those of WLH-M, whereas the content and activity of lysozyme were significantly lower. Gene Ontology functional annotation analysis and Kyoto Encyclopedia of Genes and Genomes pathway enrichment analysis revealed that immune and metabolic pathways were enriched in Vibrio-challenged clams. The expressions of the heat shock protein 70 (Hsp70) and serine protease (SP) genes, which are involved in antibacterial immunity, were significantly upregulated in WLH-M but not in ORI-M, while the expression of the kynurenine 3-monooxygenase gene, a proinflammatory factor, was significantly downregulated in WLH-M. RNA interference experiments confirmed that Hsp70 and SP downregulation could result in increased mortality of WLH-M. Therefore, we speculate that Hsp70 and SP may be involved in the antibacterial immunity of WLH-M in vivo. Our data provided a valuable resource for further studies of the antibacterial mechanism of WLH-M and provided a foundation for the breeding of pathogen-resistant strains.

Acute sulfide exposure induces hemocyte toxicity and microbiota dysbiosis in blood clam Tegillarca granosa.

Sulfide are widely accumulated in aquatic environments under anaerobic conditions, which cause health problems of aquatic animals, yet their toxic effects to benthic bivalves are not well understood. We investigated the effects of sulfide on innate immunity of the blood clam Tegillarca granosa. Immunity-related indicators and hemolymph microbiota were investigated in the clams exposed to sulfide (via 10, 100 and 1000 µmol/L of Na2S) over a 7-day period. The results showed that cellular immune responses in T. granosa were affected by exposure to high sulfide concentration (1000 µmol/L Na2S), as indicated by total counts of hemocytes (THC), cell viability, ROS levels and phagocytic activities, suggesting that sulfide stress induces T. granosa more vulnerable to pathogen challenges. In addition, the Na2S-induced stress also reshaped the hemolymph microbial community structure of T. granosa that some original genera decreased, such as Lactobacillus, Desulfovibrio and Akkermansia; some genera increased, such as Vibrio and Pseudoalteromonas in sulfide stress group. Sulfide exposure promoted the proliferation of opportunistic pathogen and reduced the diversity of microbial community in the hemolymph of T. granosa. In summary, sulfide stress had marked hemocytotoxicity, reduced immune-cell activity and increased bacterial infections in the blood clam.

Prediction and characterization of a novel hemoglobin-derived mutant peptide (mTgHbP7) from Tegillarca granosa.

The hemoglobin (Hb) is identified in Tegillarca granosa and its derived peptides have been proved to possess antibacterial activity against gram-positive and gram-negative bacteria. In this study, we identified a series of novel antimicrobial peptides (AMPs) and artificially mutated AMPs derived from subunits of T. granosa Hbs, among which, a mutant T. granosa hemoglobin peptide (mTgHbP) mTgHbP7, was proved to possess predominant antibacterial activity against three bacteria strains (Vibrio alginolyticus, V. parahaemolyticus and Escherichia coli). Besides, mTgHbP7 was predicted to form α-helical structure, which was known to be an important feature of bactericidal AMPs. Furthermore, upon contact with HEK293 cell line, we confirmed that mTgHbP7 had no cytotoxicity to mammalian cell even at a high concentration of 160 µM. Therefore, the findings reported here provide a rationalization for antimicrobial peptide prediction and optimization from mollusk hemoglobin, which will be useful for future development of antimicrobial agents.

Hypoxia-mediated immunotoxicity in the blood clam Tegillarca granosa.

In marine ecosystems, dissolved oxygen (DO) is essential for maintaining intracellular energy balance during aerobic metabolism. Bivalve mollusks are frequently exposed to hypoxia environments due to tides, temperature changes, and anthropogenic activities. The blood clam, Tegillarca granosa, mainly inhabits intertidal mudflats and is more susceptible to low oxygen events. In this study, we investigated the effect of hypoxia on immune responses in clams, and showed that hypoxia exposure reduced total hemocyte counts (THC), hemoglobin concentrations, and intracellular reactive oxygen species (ROS) levels. Also, phagocytic and cell activities of hemocyte were significantly inhibited. Furthermore, immune-related gene expression was also down-regulated. In conclusion, hypoxia greatly affected immune functions in blood clams, and our research provided the foundation for further mechanistic studies on hypoxia tolerance in clams.

Identification, characterization, and antimicrobial activity of a novel big defensin discovered in a commercial bivalve mollusc, Tegillarca granosa.

Molluscs, the second largest animal phylum on earth, primarily rely on cellular and humoral immune responses to fight against pathogen infection. Although antimicrobial peptides (AMPs) such as big defensin play crucial roles in the humoral immune response, it remains largely unknown in the ecological and economic important blood clam (Tegillarca granosa). In this study, a novel big defensin gene (TgBD) was identified in T. granosa through transcripts and whole genome searching. Bioinformatic analyses were conducted to explore the molecular characteristics of TgBD, and comparisons of TgBD with those reported in other molluscs were performed by multiple alignments and phylogenetic analysis. In addition, the expression patterns of TgBD in various tissues and upon bacterial challenge were investigated while the antimicrobial activity of synthetic N-terminal domain of TgBD was confirmed in vitro by radial diffusion experiment. Results obtained showed TgBD had an open reading frame (ORF) of 369 bp, encoding a prepropeptide containing a signal peptide and a propeptide. Similar to big defensins reported in other species, TgBD consists of a hydrophobic N-terminal domain containing ß1-α1-α2-ß2 folds and a cysteine-rich cationic C-terminal domain with three disulfide bonds between C1-C5, C2-C4, and C3-C6. Phylogenetic analysis showed that TgBD shared 76.80% similarity to its close relative ark shell (Scapharca broughtoni). In addition, TgBD expression was observed in all tissues investigated under normal conditions and was significantly induced by injection of Vibrio parahaemolyticus. Furthermore, synthetic N-terminal peptide of TgBD exhibited strong antimicrobial activity against Gram-positive bacteria tested. Our results indicated that TgBD is a constitutive and inducible acute phase AMP, which provides a universal and prompt protection for T. granosa.

Impacts of four commonly used nanoparticles on the metabolism of a marine bivalve species, Tegillarca granosa.

The rapid development of nanotechnology boosts the massive production and utilization of various nanoparticles (NPs). However, the NPs escaped into environments form emergent pollutants, which pose a potential threat to marine organisms and ecosystems. Due to their sessile filter-feeding lifestyle, marine bivalves live in pollution-prone coastal areas are more susceptible to land-sourced pollutants such as NPs. However, the impacts of many NPs on the metabolism, one of the most critical physiological processes of an organism, still remain largely unknown in bivalve species. To fill up this knowledge gap, in this study the impacts of four commonly used NPs (nZnO, nFe2O3, nCuO, and multi-walled carbon tube (MWCNT)) on the filtration rate, oxygen consumption rate, ammonia excretion rate, and O:N ratio were investigated in the blood clam, Tegillarca granosa. In addition, the expressions of ten key metabolism-related genes upon exposure to these NPs were also analyzed. The results demonstrated that exposure of blood clams to the NPs resulted in a reduction in the food intake (indicated by declined filtration rate), a shift in the metabolism substance, and disruptions in key metabolism-related molecular pathways (i.e., glycolysis and tricarboxylic acid cycle), which may render blood clam in energy shortage and thus pose significant threat to the health of this important bivalve species.

Genome-wide identification and characteristic analysis of PGRP gene family in Tegillarca granosa reveals distinct immune response of the invasive pathogen.

The peptidoglycan recognition proteins (PGRPs) are conserved innate immune molecular in invertebrates and vertebrates, which play important roles in immune system by recognize the peptidoglycans of bacterial cell walls. Although PGRPs have been extensively characterized in insects, a systematic analysis of PGRPs in bivalves is lacking. In the present study, the phylogenic relationships, gene structures and expression profiles of PGRPs in marine bivalves were analyzed. The results indicated that the most PGRPs of bivalves were predicted to degrade the peptidoglycans and prevent excessive immunostimulation of bacteria. In addition, the results of the present study showed that the protein diversity of PGRPs in most marine bivalves was mainly generated by the alternative splicing of genes, however the alternative splicing of PGRP gene family was absent in Tegillarca granosa. The differences of PGRPs might be related to the genetic and environmental differences of marine bivalves. Spatiotemporal expression profiling in T. granosa suggested that PGRPs play important roles in the immune response of invasive pathogens. The present study describes a comprehensive view of PGRPs in the blood clam T. granosa and provides a foundation for functional characterization of this gene family in innate immune of marine bivalves.

Genomic Insights into the Origin and Evolution of Molluscan Red-Bloodedness in the Blood Clam Tegillarca granosa.

Blood clams differ from their molluscan kins by exhibiting a unique red-blood (RB) phenotype; however, the genetic basis and biochemical machinery subserving this evolutionary innovation remain unclear. As a fundamental step toward resolving this mystery, we presented the first chromosome-level genome and comprehensive transcriptomes of the blood clam Tegillarca granosa for an integrated genomic, evolutionary, and functional analyses of clam RB phenotype. We identified blood clam-specific and expanded gene families, as well as gene pathways that are of RB relevant. Clam-specific RB-related hemoglobins (Hbs) showed close phylogenetic relationships with myoglobins (Mbs) of blood clam and other molluscs without the RB phenotype, indicating that clam-specific Hbs were likely evolutionarily derived from the Mb lineage. Strikingly, similar to vertebrate Hbs, blood clam Hbs were present in a form of gene cluster. Despite the convergent evolution of Hb clusters in blood clam and vertebrates, their Hb clusters may have originated from a single ancestral Mb-like gene as evidenced by gene phylogeny and synteny analysis. A full suite of enzyme-encoding genes for heme synthesis was identified in blood clam, with prominent expression in hemolymph and resembling those in vertebrates, suggesting a convergence of both RB-related Hb and heme functions in vertebrates and blood clam. RNA interference experiments confirmed the functional roles of Hbs and key enzyme of heme synthesis in the maintenance of clam RB phenotype. The high-quality genome assembly and comprehensive transcriptomes presented herein serve new genomic resources for the super-diverse phylum Mollusca, and provide deep insights into the origin and evolution of invertebrate RB.

Ocean acidification impedes gustation-mediated feeding behavior by disrupting gustatory signal transduction in the black sea bream, Acanthopagrus schlegelii.

Growing evidence suggests that ocean acidification (OA) may affect animal behaviors such as feeding. Although gustation plays a crucial role in evaluating the quality and palatability of food and ultimately influences whether or not teleosts consume the food, the potential impact of OA on gustation-mediated feeding behavior remains unknown. In this study, gustation mediated-feeding behavior, as indicated by the consumption rate (CR) and swallowing rate (SR) of agar pellets with or without feed upon OA exposure was investigated in black sea bream (Acanthopagrus schlegelii). Results showed that the exposure to acidified seawater led to significant reductions in the CR and SR of feed-containing agar pellets. In addition, the in vivo contents of three neurotransmitters and expression of genes from the gustatory signal transduction pathway were all significantly suppressed by the OA treatment. In general, the data obtained indicated that OA may hinder the gustation-mediated feeding behavior of A. schlegelii by disrupting gustatory signal transduction, which may aggravate the issue of food shortage for wild populations of black sea bream.

Immunotoxicity of microplastics and two persistent organic pollutants alone or in combination to a bivalve species.

Both microplastics and persistent organic pollutants (POPs) are ubiquitously present in natural water environment, posing a potential threat to aquatic organisms. While it has been suggested that the immune responses of aquatic organisms could be hampered by exposure to microplastics and POPs, the synergistic immunotoxic impact of these two types of pollutants remain poorly understood. In addition, little is known about the mechanism behind the immunotoxic effect of microplastics. Therefore, in the present study, the immunotoxicity of microplastics and two POPs, benzo[a]pyrene (B[a]P) and 17ß-estradiol (E2), were investigated alone or in combination in a bivalve species, Tegillarca granosa. Evident immunotoxicity, as indicated by alterations of haemocyte count, blood cell composition, phagocytic activity, intracellular content of ROS, concentration of Ca2+ and lysozyme, and lysozyme activity, was revealed for both microplastics and the two POPs examined. In addition, the expression of six immune-, Ca2+ signalling-, and apoptosis-related genes was significantly altered by exposure of clams to the contaminants studied. Furthermore, the toxicity of POPs was generally aggravated by smaller microplastics (500 nm) and mitigated by larger ones (30 µm). This size dependent effect on POP toxicity may result from size dependent interactions between microplastics and POPs. Data obtained in this study also indicate that similar to exposure to B[a]P and E2, exposure to microplastics may hamper the immune responses of clams through a series of interdependent physiological and molecular processes.

Bio-/Nanoimmobilization Platform Based on Bioinspired Fibrin-Bone@Polydopamine-Shell Adhesive Composites for Biosensing.

Inspired by blood coagulation and mussel adhesion, we report novel adhesive fibrin-bone@polydopamine (PDA)-shell composite matrix as highly efficient immobilization platform for biomacromolecules and nanomaterials. Fibrin, as a bioglue, and PDA, as a chemical adhesive, are integrated in a one-pot simultaneous polymerization consisting of biopolymerization of fibrinogen and chemical polymerization of dopamine. Fibrin fibers act as adhesive bones to construct scaffold, while PDA coat on the scaffold to form adhesive shell, generating 3D porous composite matrix with unique bone@shell structure. Two types of enzymes (glucose oxidase and acetylcholinesterase) and Au nanoparticles were adopted as respective model biomolecules and nanomaterials to investigate the immobilization capability of the matrix. The bionanocomposites showed high efficiency in capturing nanoparticles and enzymes, as well as significant mass-transfer and biocatalysis efficiencies. Therefore, the bionanocomposites exhibited significant potential in biosensing of glucose and paraoxon with limits of detection down to 5.2 µM and 4 ppt, respectively. The biological-chemical-combined polymerization strategy and composite platform with high immobilization capacity and mass-transfer efficiency open up a novel way for the preparation of high-performance bionanocomposites for various applications, in particular, biosensing.

Exposure to Waterborne nTiO2 Reduces Fertilization Success and Increases Polyspermy in a Bivalve Mollusc: A Threat to Population Recruitment.

Fertilization success is crucial for the population recruitment of an organism. However, little is known about the threat of nanoparticles (NPs) to the fertilization of broadcast spawners. Therefore, the effects of nTiO2 on fertilization success, polyspermy rate, sperm velocity, gametic DNA damage, sperm-egg collision probability, gamete fusion, and oocyte ultrastructure were investigated in a broadcast spawning bivalve, Tegillarca granosa. The results obtained show that fertilization success significantly decreased, whereas polyspermy risk markedly increased upon nTiO2 exposure. In addition, nTiO2 exposure led to a significant reduction in sperm swimming velocity, which would subsequently constrain gamete collisions. In addition, nTiO2 exposure resulted in a significant decline in gamete fusion per collision along with aggravated DNA damage in gametes. Furthermore, ultrastructural analysis illustrated the attachment of nTiO2 to the oocyte surface, which subsequently resulted in microvillus disassociation and plasma membrane damage. In conclusion, the results obtained suggest a significant threat from NP pollution to the recruitment of broadcast spawning invertebrates.

Exogenous Ca2+ mitigates the toxic effects of TiO2 nanoparticles on phagocytosis, cell viability, and apoptosis in haemocytes of a marine bivalve mollusk, Tegillarca granosa.

Phagocytosis suppression induced by nanoparticles (NPs) exposure is increasingly reported in marine species. However, the mechanisms underlying this impact remain poorly understood. In order to improve our present understanding of the immunotoxicity of NPs, acute (96 h) TiO2 NP exposure and rescue trials via exogenous supply of Ca2+ were performed in the blood clam, Tegillarca granosa. The results show that the phagocytosis rate, cell viability, and intracellular Ca2+ concentration of haemocytes were significantly suppressed, whereas the intracellular ROS concentration of haemocytes significantly increased upon nTiO2 exposure. Exposure to nTiO2 also led to the significant downregulation of Caspase-3, Caspase-6, apoptosis regulator Bcl-2, Bcl-2-associated X, calmodulin kinase II, and calmodulin kinase kinase II. Furthermore, the toxic impacts of nTiO2 were partially mitigated by the addition of exogenous Ca2+, as indicated by the recovery tendency in almost all the measured parameters. The present study indicates that Ca2+ signaling could be one of the key pathways through which nTiO2 attacks phagocytosis.

Immunotoxicity of four nanoparticles to a marine bivalve species, Tegillarca granosa.

The increasing application of nanomaterials drives the unintentional release of nanoparticles (NPs) into the ocean, which may pose a potential threat to marine organisms. It has been demonstrated that exposure to NPs could chanllenge the immune responses of marine species. However, the affecting mechanism behind remains poorly understood. In this study, the immunotoxic impacts and the mechanisms underpinning the effects of four major NPs, including nZnO, nFe2O3, nCuO, and carbon nanotube (MWCNT), were investigated in blood clam, Tegillarca granosa. The results showed that exposure to tested NPs resulted in reduced total counts, altered cell composition, and constrained phagocytic activities of haemocytes. The intracellular contents of reactive oxygen species (ROS) and the degree of DNA damage of haemocytes were significantly induced, whereas the haemocyte viability was suppressed. Furthermore, NP exposures led to significant increases in the in vivo contents of neurotransmitters. Down-regulations of the immune- and neurotransmitter-related genes were detected as well. Our data suggest that NP exposures hampered the immune responses of blood clams most likely through (1) inducing ROS, causing DNA damage, and reducing cell viability of haemocytes, (2) altering the in vivo contents of neurotransmitters, and (3) affecting the expression of immune- and neurotransmitter-related genes.

Fluoxetine suppresses the immune responses of blood clams by reducing haemocyte viability, disturbing signal transduction and imposing physiological stress.

The antidepressant fluoxetine (FLX), a selective serotonin reuptake inhibitor, is widely prescribed for the treatment of depression and anxiety disorders. Nowadays, measurable quantities of FLX have been frequently detected in the aquatic ecosystems worldwide, which may pose a potential threat to aquatic organisms. Although the impacts of FLX exposure on immune responses are increasingly well documented in mammals, they remain poorly understood in aquatic invertebrates. Therefore, to gain a better understanding of the ecotoxicological effects of FLX, the impacts of waterborne FLX exposure on the immune responses of blood clam, Tegillarca granosa, were investigated in this study. Results obtained showed that both cellular and humoural immune responses in T. granosa were suppressed by exposure to waterborne FLX, as indicated by total counts of haemocytes (THC), phagocytic rate, and activities of superoxide dismutases (SOD) and catalase (CAT), suggesting that waterborne FLX renders blood clams more vulnerable to pathogen challenges. To ascertain the mechanisms explaining how waterborne FLX affects immune responses, haemocyte viabilities, intracellular Ca2+ levels, in vivo concentrations of neurotransmitters, physiological stress conditions (as indicated by in vivo concentrations of cortisol), and expressions of key regulatory genes from Ca2+ and neurotransmitter signal transduction, as well as immune-related signalling pathways, were examined after 10 days of FLX exposure by blood clams via 1, 10 and 100 µg/L waterborne FLX. The results obtained indicated that immune response suppression caused by waterborne FLX could be due to (i) inhibited haemocyte viabilities, which subsequently reduce the THC; (ii) altered intracellular Ca2+ and neurotransmitter concentrations, which lead to constrained phagocytosis; and (iii) aggravated physiological stress, which thereafter hampers immune-related NFκB signalling pathways.

Anthropogenic Noise Aggravates the Toxicity of Cadmium on Some Physiological Characteristics of the Blood Clam Tegillarca granosa.

Widespread applications of cadmium (Cd) in various products have caused Cd contamination in marine ecosystems. Meanwhile, human activities in the ocean have also generated an increasing amount of noise in recent decades. Although anthropogenic noise and Cd contaminants could be present simultaneously in marine environments, the physiological responses of marine bivalve mollusks upon coexposure to anthropogenic noise and toxic metal contaminants, including Cd remain unclear. Therefore, the combined effects of anthropogenic noise and Cd on the physiological characteristics of the blood clam Tegillarca granosa were investigated in this study. The results showed that 10 days of coexposure to anthropogenic noise and Cd can enhance adverse impacts on metabolic processes, as indicated by the clearance rate, respiration rate, ammonium excretion rate, and O:N ratio of T. granosa. In addition, both the ATP content, ATP synthase activity and genes encoding important enzymes in ATP synthesis significantly declined after coexposures to anthropogenic noise and Cd, which have resulted from reduced feeding activity and respiration. Furthermore, the expressions of neurotransmitter-related genes (MAO, AChE, and mAChR3) were all significantly down-regulated after coexposure to anthropogenic noise and Cd, which suggests an enhanced neurotoxicity under coexposure. In conclusion, our study demonstrated that anthropogenic noise and Cd would have synergetic effects on the feeding activity, metabolism, and ATP synthesis of T. granosa, which may be due to the add-on of stress responses and neurotransmitter disturbances.