Immune responses of hemocytes in the blood clam Tegillarca granosa in response to in vivo Vibrio harveyi infection.

Aquaculture of the blood clam Tegillarca granosa accounts for approximately 50% of Arcidae (ark shell) production in China. Vibrio infection severely threatens the sustainability of the clam aquaculture industry. Exposure to Vibrio induces an immune response in blood clams. However, the underlying mechanism remains poorly understood. In this study, immune responses of hemocytes in blood clams were detected after Vibrio infection; the immersion method was used in vivo to mimic the clam's natural infection process. After 24 h of exposure to Vibrio infection, the Vibrio load in hemolymph fluid in both the treatment Ⅰ (25,033.33 ± 19,563.11 CFU/mL) and treatment Ⅱ (122,163.33 ± 194,409.49 CFU/mL) groups were significantly higher, than that in the control group (13.67 ± 37.73 CFU/mL) (P < 0.05). Correspondingly, the production of intracellular reactive oxygen species was approximately 1.40 (treatment Ⅰ) and 2.12 (treatment Ⅱ) fold higher than that in the control group (P < 0.05), and the induced DNA damage showed a similar trend (P < 0.05). Vibrio infection also significantly increased lysozyme content, adenosine triphosphate content, and peroxidase isozyme activity, in both the serum and hemocyte lysates (P < 0.05). The expression of immune-associated genes (ABCA3, c-Myc, Caspase 3, and HSP70) was upregulated under infection conditions. The phagocytic activity was approximately 1.99 (treatment Ⅰ) and 2.57 (treatment Ⅱ) fold that in control clams (P < 0.05). In addition, the total hemocyte count and red granulocyte percentage both significantly decreased by approximately 75-90% after Vibrio infection. These results provided novel insights into the mechanism of hemocyte immunity in T. granosa against Vibrio infection, which may aid in the future prevention and control of Vibrio infection in vivo.

An Efficient CRT Based Algorithm for Frequency Determination from Undersampled Real Waveform.

The Chinese Remainder Theorem (CRT) based frequency estimation has been widely studied during the past two decades. It enables one to estimate frequencies by sub-Nyquist sampling rates, which reduces the cost of hardware in a sensor network. Several studies have been done on the complex waveform; however, few works studied its applications in the real waveform case. Different from the complex waveform, existing CRT methods cannot be straightforwardly applied to handle a real waveform's spectrum due to the spurious peaks. To tackle the ambiguity problem, in this paper, we propose the first polynomial-time closed-form Robust CRT (RCRT) for the single-tone real waveform, which can be considered as a special case of RCRT for arbitrary two numbers. The time complexity of the proposed algorithm is O(L), where L is the number of samplers. Furthermore, our algorithm also matches the optimal error-tolerance bound.

Characterization of a novel activating protein-1 (AP-1) gene and the association of its single nucleotide polymorphisms with vibrio resistance in Tegillarca granosa.

The blood clam Tegillarca granosa is a commercial marine bivalve of economic value, accounting for approximately 50% of clam production in China. In recent years, the yield of blood clams has been threatened by bacterial infections caused by marine Vibrio species that thrive under a rising sea temperature. The transcription factor activating protein-1 (AP-1) is emerging as an important player in the innate immunity of marine bivalves against viral or bacterial infections. In this study, the full-length cDNA of a novel T. granosa AP-1 (TgAP-1) was cloned for the first time. The 1591-bp cDNA encoded a protein of 292 amino acid residues with a calculated molecular weight of 32.8 kDa. The TgAP-1 protein contained an N-terminal Jun domain and a C-terminal basic region leucine zipper domain typically found in Jun proteins (a subfamily of AP-1 proteins). TgAP-1 was ubiquitously expressed in T. granosa, with the highest expression detected in the gill and foot, followed by the mantle, hemolymph, and hepatopancreas. Exposure to Vibrio harveyi induced TgAP-1 expression in gill tissues and the expression levels of TgAP-1 of resistant blood clams were always lower than that of control population whether Vibro infection or not. A total of 18 single nucleotide polymorphisms (SNPs) of TgAP-1 were detected in T. granosa. SNP-typing and haplotyping of resistant and susceptible populations revealed that six SNPs (AG type of TgSNP-1, GA type of TgSNP-2, TG type of TgSNP-4, CT type of TgSNP-7, AG type of TgSNP-11, and GA type of TgSNP-12) and four haplotypes (fHap2, fHap3, fHap6, and fHap7) were significantly associated with V. harveyi resistance. Risk assessment showed that fHap2 (CG) and fHap7 (GA) were associated with an increased resistance, while fHap3 (CT) and fHap6 (AG) were associated with an increased susceptibility. The results from this study supported a potential role of TgAp-1 in the anti-Vibro immunity of T. granosa. The discovery of the genetic molecular markers and haplotypes related to Vibrio resistance can provide guidance for selective breeding of T. granosa in the future.

Bacterial Community Dynamics During Nursery Rearing of Pacific White Shrimp (Litopenaeus vannamei) Revealed via High-Throughput Sequencing.

A 20-day trial was conducted to reveal bacterial community dynamics in a commercial nursery of larval Litopenaeus vannamei larvae. The bacterial communities in the ambient water were profiled by high-throughput sequencing of the V4-V5 hypervariable region of the 16S rRNA gene. The results indicated that the dominant bacterial phyla between the metamorphosis stage and postlarval stage were Bacteroidetes, Proteobacteria, Cyanobacteria, and Firmicutes, representing more than 80.09% of the bacterial operational taxonomic units. The relative abundance among bacterial phyla notably differed between the two stages. The relative abundance of Cyanobacteria was higher in the metamorphosis stage, while that of Bacteroidetes was higher and more stable in the postlarval stage. At the class level, the relative abundance of Sphingobacteriia and Alphaproteobacteria increased markedly in the postlarval stage, while that of Flavobacteriia decreased. Redundancy analysis showed that bacterial composition in the metamorphosis stage was positively correlated with salinity, alkalinity, and pH, while in the postlarval stage, it was positively correlated with ammonium nitrogen and nitrite nitrogen. Thus, microbial community diversity in the nursery phase varies per rearing stage.

The synergic impacts of TiO2 nanoparticles and 17ß-estradiol (E2) on the immune responses, E2 accumulation, and expression of immune-related genes of the blood clam, Tegillarca granosa.

The extensive use of TiO2 nanoparticles (nTiO2) in industrial products has led to their release into the marine environment, thereby posing a potential risk to marine organisms. However, in addition to affecting marine organisms through its inherent properties, nTiO2 can also act as a vehicle for other toxic pollutants due to their strong adsorption ability through the "Trojan horse" effect. Due to their potential hazard, the endocrine disrupting chemicals (EDCs) such as 17ß-estradiol (E2), have been considered as one of the most serious anthropogenic threats to biodiversity and ecosystem health. However, there is still a lack of knowledge regarding the possible synergistic effects of nTiO2 and endocrine disrupting chemicals (EDCs) on marine organisms to date. Therefore, the combined effects of nTiO2 and 17ß-estradiol (E2) on the immune responses of the blood clam, Tegillarca granosa, were investigated in this study. After 10 days of treatment, the total number, phagocytic activity, red granulocytes ratio, and the phagocytosis of hemocytes were significantly reduced in almost all treatment groups. Furthermore, expressions of genes from NFκß and Toll-like receptor signaling pathways were significantly altered after exposure to nTiO2 and/or E2, indicating a reduced sensitivity to pathogen challenges. In addition, compared to exposure to E2 alone, co-exposure to E2 and nTiO2 led to a significant increase in the content of alkali-labile phosphate (ALP) in hemolymph, suggesting an enhanced E2 bioconcentration in the presence of nTiO2. In general, the present study demonstrated that nTiO2 enhanced the immunotoxicity of E2 to the blood clam, which may be due to the increased E2 uptake in the presence of nTiO2.

Laboratory simulation reveals significant impacts of ocean acidification on microbial community composition and host-pathogen interactions between the blood clam and Vibrio harveyi.

It has been suggested that climate change may promote the outbreaks of diseases in the sea through altering the host susceptibility, the pathogen virulence, and the host-pathogen interaction. However, the impacts of ocean acidification (OA) on the pathogen components of bacterial community and the host-pathogen interaction of marine bivalves are still poorly understood. Therefore, 16S rRNA high-throughput sequencing and host-pathogen interaction analysis between blood clam (Tegillarca granosa) and Vibrio harveyi were conducted in the present study to gain a better understanding of the ecological impacts of ocean acidification. The results obtained revealed a significant impact of ocean acidification on the composition of microbial community at laboratory scale. Notably, the abundance of Vibrio, a major group of pathogens to many marine organisms, was significantly increased under ocean acidification condition. In addition, the survival rate and haemolytic activity of V. harveyi were significantly higher in the presence of haemolymph of OA treated T. granosa, indicating a compromised immunity of the clam and enhanced virulence of V. harveyi under future ocean acidification scenarios. Conclusively, the results obtained in this study suggest that future ocean acidification may increase the risk of Vibrio pathogen infection for marine bivalve species, such as blood clams.

Benzo[a]pyrene exposure under future ocean acidification scenarios weakens the immune responses of blood clam, Tegillarca granosa.

Persistent organic pollutants (POPs) are known to converge into the ocean and accumulate in the sediment, posing great threats to marine organisms such as the sessile bottom burrowing bivalves. However, the immune toxicity of POPs, such as B[a]P, under future ocean acidification scenarios remains poorly understood to date. Therefore, in the present study, the impacts of B[a]P exposure on the immune responses of a bivalve species, Tegillarca granosa, under present and future ocean acidification scenarios were investigated. Results obtained revealed an increased immune toxicity of B[a]P under future ocean acidification scenarios in terms of reduced THC, altered haemocyte composition, and hampered phagocytosis, which may attribute to the synergetic effects of B[a]P and ocean acidification. In addition, the gene expressions of pathogen pattern recognition receptors (TLR1, TLR2, TLR4, TLR6), pathway mediators (TRAF6, TAK1, TAB2, IKKα and Myd88), and effectors (NF-ĸB) of the important immune related pathways were significantly down-regulated upon exposure to B[a]P under future ocean acidification scenarios. Results of the present study suggested an increased immune toxicity of B[a]P under future ocean acidification scenarios, which will significantly hamper the immune responses of T. granosa and subsequently render individuals more susceptible to pathogens challenges.

Log-less metadata management on metadata server for parallel file systems.

This paper presents a novel metadata management mechanism on the metadata server (MDS) for parallel and distributed file systems. In this technique, the client file system backs up the sent metadata requests, which have been handled by the metadata server, so that the MDS does not need to log metadata changes to nonvolatile storage for achieving highly available metadata service, as well as better performance improvement in metadata processing. As the client file system backs up certain sent metadata requests in its memory, the overhead for handling these backup requests is much smaller than that brought by the metadata server, while it adopts logging or journaling to yield highly available metadata service. The experimental results show that this newly proposed mechanism can significantly improve the speed of metadata processing and render a better I/O data throughput, in contrast to conventional metadata management schemes, that is, logging or journaling on MDS. Besides, a complete metadata recovery can be achieved by replaying the backup logs cached by all involved clients, when the metadata server has crashed or gone into nonoperational state exceptionally.

Association among attention-deficit hyperactivity disorder, restless legs syndrome, and peripheral iron status: a two-sample Mendelian randomization study.

Background: Epidemiological evidence indicates a high correlation and comorbidity between Attention Deficit Hyperactivity Disorder (ADHD) and Restless Legs Syndrome (RLS). Objective: We aimed to investigate the causal relationship and shared genetic architecture between ADHD and RLS, as well as explore potential causal associations between both disorders and peripheral iron status. Methods: We performed two-sample Mendelian randomization (MR) analyses using summary statistics from genome-wide meta-analyses of ADHD, RLS, and peripheral iron status (serum iron, ferritin, transferrin saturation, and total iron binding capacity). Additionally, we employed linkage disequilibrium score regression (LDSC) to assess genetic correlations between ADHD and RLS using genetic data. Results: Our MR results supports a causal effect from ADHD (as exposure) to RLS (as outcome) (inverse variance weighted OR = 1.20, 95% CI: 1.08-1.34, p = 0.001). Conversely, we found no a causal association from RLS to ADHD (inverse variance weighted OR = 1.04, 95% CI: 0.99-1.09, p = 0.11). LDSC analysis did not detect a significant genetic correlation between RLS and ADHD (Rg = 0.3, SE = 0.16, p = 0.068). Furthermore, no evidence supported a causal relationship between peripheral iron deficiency and the RLS or ADHD onset. However, RLS may have been associated with a genetic predisposition to reduced serum ferritin levels (OR = 1.20, 95% CI: 1.00-1.04, p = 0.047). Conclusion: This study suggests that ADHD is an independent risk factor for RLS, while RLS may confer a genetic predisposition to reduced serum ferritin levels. Limitations: The GWAS summary data utilized originated from populations of European ancestry, limiting the generalizability of conclusions to other populations. Clinical implications: The potential co-occurrence of RLS in individuals with ADHD should be considered during diagnosis and treatment. Moreover, iron supplementation may be beneficial for alleviating RLS symptoms.

Nomogram-Based Prediction of the Futile Recanalization Risk Among Acute Ischemic Stroke Patients Before and After Endovascular Therapy: A Retrospective Study.

Background and Purpose: Futile recanalization (FRC) is common among large artery occlusion (LAO) patients after endovascular therapy (EVT). We developed nomogram models to identify LAO patients at a high risk of FRC pre- and post-EVT to help neurologists select the optimal candidates for EVT. Methods: From April 2020 to July 2022, EVT and mTICI score ≥2b LAO patients were recruited. Nomogram models was developed by two-step approach for predicting the outcomes of LAO patients. First, the least absolute shrinkage and selection operator (LASSO) regression analysis was to optimize variable selection. Then, a multivariable analysis was to construct an estimation model with significant indicators from the LASSO. The accuracy of the model was verified using receiver operating characteristic (ROC), calibration curve, and decision curve analyses (DCA), along with validation cohort (VC). Results: Using LASSO, age, sex, hypertension history, baseline NIHSS, ASPECTS and baseline SBP upon admission were identified from the pre-EVT variables. Model 1 (pre-EVT) showed good predictive performance, with an area under the ROC curve (AUC) of 0.815 in the training cohort (TrC) and 0.904 in VC. Under the DCA, the generated nomogram was clinically applicable where risk cut-off was between 15%-85% in the TrC and 5%-100% in the VC. Moreover, age, ASPECTS upon admission, onset duration, puncture-to-recanalization (PTR) duration, and lymphocyte-to-monocyte ratio (LMR) were screened by LASSO. Model 2 (post-EVT) also demonstrated good predictive performance with AUCs of 0.888 and 0.814 for TrC and VC, respectively. Under the DCA, the generated nomogram was clinically applicable if the risk cut-off was between 13-100% in the TrC and 22-85% of VC. Conclusion: In this study, two nomogram models were generated that showed good discriminative performance, improved calibration, and clinical benefits. These nomograms can potentially accurately predict the risk of FRC in LAO patients pre- and post-EVT and help to select appropriate candidates for EVT.

Image Synthesis and Modified BlendMask Instance Segmentation for Automated Nanoparticle Phenotyping.

Automated nanoparticle phenotyping is a critical aspect of high-throughput drug research, which requires analyzing nanoparticle size, shape, and surface topography from microscopy images. To automate this process, we present an instance segmentation pipeline that partitions individual nanoparticles on microscopy images. Our pipeline makes two key contributions. Firstly, we synthesize diverse and approximately realistic nanoparticle images to improve robust learning. Secondly, we improve the BlendMask model to segment tiny, overlapping, or sparse particle images. Specifically, we propose a parameterized approach for generating novel pairs of single particles and their masks, encouraging greater diversity in the training data. To synthesize more realistic particle images, we explore three particle placement rules and an image selection criterion. The improved one-stage instance segmentation network extracts distinctive features of nanoparticles and their context at both local and global levels, which addresses the data challenges associated with tiny, overlapping, or sparse nanoparticles. Extensive experiments demonstrate the effectiveness of our pipeline for automating nanoparticle partitioning and phenotyping in drug research using microscopy images.

Toxicity assessment and detoxification metabolism of sodium pentachlorophenol (PCP-Na) on marine economic species: a case study of Moerella iridescens and Exopalaemon carinicauda.

Sodium pentachlorophenol (PCP-Na) is widespread in the marine environment; however, its impact on marine organisms remains under-researched. Moerella iridescens and Exopalaemon carinicauda are marine species of economic importance in China and under threat from PCP-Na pollution. Thus, this study aimed to assess the toxicity and detoxification metabolism of PCP-Na on M. iridescens and E. carinicauda. The study revealed that the 96 h median lethal concentration (LC50) of PCP-Na for M. iridescens and E. carinicauda were 9.895 mg/L and 14.143 mg/L, respectively. A species sensitivity distribution (SSD) for PCP-Na was developed specifically for marine organisms, determining a hazardous concentration to 5% of the species (HC5) of 0.047 mg/L. During the sub-chronic exposure period, PCP-Na accumulated significantly in M. iridescens and E. carinicauda, with highest concentrations of 41.22 mg/kg in the soft tissues of M. iridescens, 42.58 mg/kg in the hepatopancreas of E. carinicauda, and only 0.85 mg/kg in the muscle of E. carinicauda. Furthermore, the study demonstrated that detoxifying metabolic enzymes and antioxidant defense system enzymes of E. carinicauda responded stronger to PCP-Na compared to M. iridescens, suggesting that E. carinicauda may possess a stronger detoxification capacity. Notably, five biomarkers were identified and proposed for monitoring and evaluating PCP-Na contamination. Overall, the results indicated that M. iridescens and E. carinicauda exhibit greater tolerance to PCP-Na than other marine species, but they are susceptible to accumulating PCP-Na in their tissues, posing a significant health risk. Consequently, conducting aquatic health risk assessments in areas with potential PCP-Na contamination is strongly recommended.

Study on the toxic effects of sodium pentachlorophenol (PCP-Na) on razor clam (Sinonovacula constricta).

Currently, research on toxic effects of PCP Na is greatly insufficient. The aim of this study is to explore the toxic effects of PCP-Na for better conducting future work on PCP-Na toxicology. For this purpose, S. constricta adults were exposed to PCP-Na for toxicity testing. The results showed that PCP-Na could easily bioaccumulate in S. constricta and significantly affected both phrase I and II metabolism enzymes. Meanwhile, PCP-Na strongly activated antioxidant system and caused PC, LPO and DNA damage. In addition, neurotoxicity and immunotoxicity of PCP-Na was demonstrated in this study. Interestingly, we observed that PCP-Na significantly affected the expression of genes of electron transport chain and induced key enzymes of glycolysis, indicating that PCP-Na may act as an uncoupler of oxidative phosphorylation, interfering with energy supply and causing energy compensation. This study is the first to fully analyze and provide a new perspective on the toxicity of PCP-Na.

3-Nitro-2H-chromenes as a new class of inhibitors against thioredoxin reductase and proliferation of cancer cells.

A series of 3-nitrochromenes were designed and synthesized. These compounds showed good inhibitory activity against thioredoxin reductase (TrxR) and the proliferation of A549 cancer cells. The structure-activity relationship analysis indicates that the 3-nitrochromene scaffold is the crucial pharmacophore for achieving good inhibitory activity. The bromo-substitutions at the 6- and 8-position of 3-nitrochromene significantly increase the inhibitory activity.

Expression of ABCA3 transporter gene in Tegillarca granosa and its association with cadmium accumulation.

Exposure to cadmium (Cd), a heavy metal, can cause strong and toxic side effects. Cd can enter the body of organisms in several ways, leading to various pathological reactions in the body. Tegillarca granosa is a kind of bivalve shellfish favored by people in the coastal areas of China. Bivalve shellfish can easily absorb heavy metal pollutants from water bodies while filter feeding. T. granosa is considered a hyper-accumulator of Cd, and the TgABCA3 gene is highly expressed in individuals with a high content of Cd-exposed blood clam. However, it is unclear whether TgABCA3 is involved in Cd ion transport in blood clam and the molecular mechanism for the mechanism of the Cd-induced responses for maintaining cell homeostasis. In this study, the complete cDNA of the TgABCA3 gene was analyzed to provide insights into the roles of TgABCA3 in resistance against Cd in blood clam. The complete sequence of TgABCA3 showed high identity to that of TgABCA3 from other bivalves and contained some classical motifs of ATP-binding cassette transport proteins. TgABCA3 expression in different tissues was measured using real-time quantitative polymerase chain reaction (qRT-PCR) and western blot analysis. The tissue-specific expression showed that TgABCA3 expression was highest in the gill tissue. The TgABCA3 expression in the gill tissue was silenced using the RNA interference technique. After TgABCA3 silencing, the TgABCA3 expression decreased, the Cd content increased, the oxygen consumption and ammonia excretion rates increased, and the ingestion rate decreased. These results showing that the extents of Cd accumulation and resulting toxic effects are related to expression levels and activity of TgABCA3 indicate that TgABCA3 has a protective function against Cd in the clam. This increase in Cd accumulation results in serious damage to the body, leading to the enhancement of its physiological metabolism. Therefore, the findings of the study demonstrated that TgABCA3 can participate in the transport of Cd ions in the blood clam through active transport and play a vital role in Cd detoxification.

Molecular cloning, characterization, and tissue distribution of c-Myc from blood clam Tegillarca granosa and its role in cadmium-induced stress response.

Cadmium (Cd) pollution threatens the cultivation of the blood clam Tegillarca granosa (T. granosa) in coastal regions of the East China Sea. The molecular mechanisms regulating Cd stress response and detoxification in blood clams are largely unclear. In the present study, the full-length T. granosa c-Myc (Tgc-Myc) cDNA was cloned for the first time. The 3063-bp cDNA consisted of a 129-bp 5' untranslated region (UTR), a 1746-bp 3' UTR, and a 1188-bp open reading frame encoding a predicted protein of 395 amino acid residues. The predicted protein had a calculated molecular weight of 44.9 kDa and an estimated isoelectric point of 6.82. The predicted protein contained an N-terminal transactivation domain and a C-terminal basic helix-loop-helix leucine zipper domain, which are conserved functional domains of c-Myc proteins. Tgc-Myc showed broad tissue distribution in blood clams, with the highest expression detected in the gill and hepatopancreas. Exposure to Cd, a major heavy metal pollutant in coastal regions of the East China Sea, induced Tgc-Myc expression in gill tissues. Tgc-Myc knockdown led to reduced expression of a variety of stress response/detoxification genes in blood clams cultivated in Cd-contaminated seawater. Tgc-Myc knockdown also led to decreased expression of IGF1R, a proto-oncogene that promotes cell proliferation. These findings indicated that Tgc-Myc regulates Cd-induced stress response and detoxification in blood clams. The upregulation of Tgc-Myc may serve as an approach to generate strains with an enhanced detoxification response and consequently a low heavy metal buildup.

Microplastics boost the accumulation of tetrabromobisphenol A in a commercial clam and elevate corresponding food safety risks.

Marine bivalve molluscs are one of the primary seafood for consumers. Inhabiting terrigenous pollutant-convergent coastal areas and feeding through seawater filtration, edible bivalves are subjected to waterborne emerging pollutants such as microplastics (MPs) and tetrabromobisphenol A (TBBPA). Nevertheless, the potential risks of consuming MP-TBBPA contaminated seafood are still largely unknown. With that, accumulation of TBBPA with and without the presence of MPs in a commercial bivalve species, blood clam (Tegillarca granosa), was determined in the present study. Meanwhile, corresponding target hazard quotients (THQs) as well as margins of exposure (MoEs) were estimated to evaluate the potential health risks for clam consumers. Furthermore, the impacts of pollutants accumulation on the detoxification process and energy supply were analysed. The data obtained demonstrated that MPs aggravate the accumulation of TBBPA in clams, leading to elevated potential food safety risks (indicated by higher THQ values and lower MoE values) for consumers. In addition, the in vivo contents of CYP1A1 and UDP-glucuronosyltransferase, the enzymatic activity of glutathione-S-transferase, and the expression levels of five detoxification-related genes were all dramatically suppressed by MP-TBBPA. Furthermore, clams exposed to MP-TBBPA had significantly lower adenosine triphosphate contents and lower pyruvate kinase and phosphofructokinase activities. These results indicated that the aggravation of TBBPA accumulation may be due to the hence disruption of detoxification process and limited energy available for detoxification.

Microplastics aggravate the bioaccumulation of three veterinary antibiotics in the thick shell mussel Mytilus coruscus and induce synergistic immunotoxic effects.

Marine bivalves living in pollution-prone coastal areas may be simultaneously coexposed to veterinary antibiotic residuals and microplastics (MPs). However, the synergistic effects of these two types of emergent pollutants have not been fully elucidated in mussel species. Therefore, the immunotoxic effects of the three representative antibiotics, oxytetracycline (OTC, 270 ng/L), florfenicol (FLO, 42 ng/L), and sulfamethoxazole (SMX, 140 ng/L), with and without the copresence of polystyrene MPs (0.26 mg/L, dimeter: 500 nm), were investigated in the thick shell mussel. Our data showed that the immunity was significantly hampered by exposure to the pollutants and MP-antibiotic coexposure induced synergistic immunotoxicity. For instance, compared to those treated with antibiotics (OTC, FLO, and SMX) alone, mussels coexposed to antibiotic and MPs had significantly lower phagocytic rate (further decline by approximately 28.80%, 34.21%, and 11.22%, respectively) and total hemocyte count (further reduced by approximately 37.45%, 61.67%, and 46.32%, respectively). Exposure to the pollutants tested also led to inductions in intracellular reactive oxygen species (ROS), decreases in the F-actin cytoskeleton, declines in the cell viability of hemocytes, and downregulation of cytoskeleton- and immune-related genes. In addition, mussels coexposed to antibiotic-MP accumulated significantly greater amounts of antibiotics, which may partially explain the synergic immunotoxic effect detected. Exposure to pollutants tested also led to suppression in the activity of glutathione-S-transferase (GST) and downregulation of detoxification-related genes whereas induction in the level of lipid peroxidation (indicated by MDA content) in gills, which may facilitate the entry whereas constrain the exclusion of antibiotics and therefore result in an elevation in accumulation of antibiotics.

Transcriptome analysis of sex-related genes in the blood clam Tegillarca granosa.

BACKGROUND: Blood clams (Tegillarca granosa) are one of the most commercial shellfish in China and South Asia with wide distribution in Indo-Pacific tropical to temperate estuaries. However, recent data indicate a decline in the germplasm of this species. Furthermore, the molecular mechanisms underpinning reproductive regulation remain unclear and information regarding genetic diversity is limited. Understanding the reproductive biology of shellfish is important in interpreting their embryology development, reproduction and population structure. Transcriptome sequencing (RNA-seq) rapidly obtains genetic sequence information from almost all transcripts of a particular tissue and currently represents the most prevalent and effective method for constructing genetic expression profiles. RESULTS: Non-reference RNA-seq, an Illumina HiSeq2500 Solexa system, and de novo assembly were used to construct a gonadal expression profile of the blood clam. A total of 63.75 Gb of clean data, with at least 89.46% of Quality30 (Q30), were generated which was then combined into 214,440 transcripts and 125,673 unigenes with a mean length of 1,122.63 and 781.30 base pairs (bp). In total, 27,325 genes were annotated by comparison with public databases. Of these, 2,140 and 2,070 differentially expressed genes (DEGs) were obtained (T05 T08 vs T01 T02 T04, T06 T07 vs T01 T02 T04; in which T01-T04 and T05-T08 represent biological replicates of individual female and male clams, respectively) and classified into two groups according to the evaluation of biological replicates. Then 35 DEGs and 5 sex-related unigenes, in other similar species, were investigated using qRT-PCR, the results of which were confirmed to data arising from RNA-seq. Among the DEGs, sex-related genes were identified, including forkhead box L2 (Foxl2), sex determining region Y-box (Sox), beta-catenin (ß-catenin), chromobox homolog (CBX) and Sex-lethal (Sxl). In addition, 6,283 simple sequence repeats (SSRs) and 614,710 single nucleotide polymorphisms (SNPs) were identified from the RNA-seq results. CONCLUSIONS: This study provided the first complete gonadal transcriptome data for the blood clam and allowed us to search many aspects of gene sequence information, not limited to gender. This data will improve our understanding of the transcriptomics and reproductive biology of the blood clam. Furthermore, molecular markers such as SSRs and SNPs will be useful in the analysis of genetic evolution, bulked segregant analysis (BSA) and genome-wide association studies (GWAS). Our transcriptome data will therefore provide important genetic information for the breeding and conservation of germplasm.

Transcriptome analysis of shell color-related genes in the clam Meretrix meretrix.

Color polymorphism has received much attention due to its strong implications for speciation and adaptation. In contrast to body color, little is currently known about the molecular mechanism of shell color formation. This study represents the first analysis of the relationship between whole-scale gene expression and shell color variations in the marine bivalve mollusks via comparative transcriptome analyses. Three clam Meretrix meretrix strains with different and monotonous shell color patterns, which were developed by our 10-year artificial selection, combined with clams with nearly white shell color were used in the analyses. The results supported the idea that there was a relationship between gene expression and shell pigmentation in the clam M. meretrix, and complex signal transduction were involved. It was proposed that Notch signaling pathway played a crucial role in shell pigmentation in a gene-dosage dependent pattern and also potentially involved in the shell color patterning. Calcium signaling process may equally be implicated in shell color formation via activation of Notch pathway. Other differentially expressed genes (e.g., Myl, Mitf) potentially implicated in shell color pigmentation were also noticed. This study provides information on the expression profiles of clams with different shell color morphs and sheds light on color formation mechanism of shell.