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The ichthyotoxic raphidophyte Heterosigma akashiwo is associated with harmful algal blooms (HABs). Although the influence of H. akashiwo on fish has been reported, little is known of its effects on other marine organisms, such as shellfish. In this study, the physiological and transcriptomic responses of the Asiatic hard clam Meretrix meretrix to H. akashiwo were investigated. The results showed that clam survival was not influenced by H. akashiwo (at a concentration of 2 × 104 cells/mL), and M. meretrix eliminated the alga effectively after 48 h in co-culture. H. akashiwo did not alter the antioxidant capacity of the clams. However, 544 unigenes were found to be differentially expressed between the control and H. akashiwo-treated clams. Interestingly, the significant up-regulation of lysosome-related genes and transport proteins suggested remodeling of the clam metabolism possibly for digestion of H. akashiwo. In summary, our results indicate that the clam M. meretrix can effectively feed on H. akashiwo, and intensification of shellfish farming may be helpful to control and mitigate H. akashiwo blooms. [ABSTRACT FROM AUTHOR]

Simple Summary: Meretrix meretrix lives in substrates at a depth of 1–20 cm; the substrate has an influence on its growth, survival, living habits, and behavioral characteristics. In this study, we investigated the effects of different grain size substrates on substrate preference, burrowing ability, and behavior during the substrate selection process of M. meretrix. These results indicated that juvenile M. meretrix had a significant preference for and a stronger burrowing ability in fine sand. As the substrate grain size increased, the burrowing ability and preference of M. meretrix decreased, and these bivalves showed behavioral characteristics such as a prolonged selection time and an increased percentage of movement. In addition, by observing the substrate selection behavior of M. meretrix, we divided the selection process of the substrate by juvenile M. meretrix into four stages: preparation, selection, burrowing, and end stages. The substrate is the key environmental factor that affects the growth, survival, population and distribution of dwelling mollusks in mudflat settings. To clarify the effect of the substrate grain size on soft substrate preference, burrowing ability and behavior during the selection process of juvenile Meretrix meretrix, four different grain size substrates (coarse sand, medium sand, fine sand, and natural substrate) were set up for comparison. The results indicated that: (1) the burrowing ability of juvenile specimens in fine sand was the strongest; (2) the degree (from high to low) of the juvenile's preference for the four substrates was in the order of fine sand > natural substrate > medium sand > coarse sand; and (3) the selection process of the substrate by the juveniles could be divided into four stages: preparation, selection, burrowing and end stages. These stages showed the behavioral characteristics of a longer selection time and higher percentage of movement in coarse sand. Therefore, our results demonstrated that sea areas or ponds with fine sand as the main component are more suitable for stock enhancement with M. meretrix. These results provide basic data for habitat selection and suitability evaluations for the aquaculture of M. meretrix. [ABSTRACT FROM AUTHOR]

Despite mass mortality, hard clams remain among the main cultured shellfish in Taiwan. Using cross-sectional data, this study applies threshold regression modeling with stocking density and feed costs as the threshold variables to explore the nonlinear relationship between input and output factors. The findings show that the production output levels may be increased by different combinations of factor inputs and variations in input factors. More specifically, the higher output levels may be achieved by increasing labor input while reducing capital input factors in farming households with a higher stocking density (HSD > 1,087,870 inds/ha) or higher feed cost (HFC > 13,889 NTD/ha). Farming households with a lower stocking density (LSD ≤ 1,087,870 inds/ha) may enhance production output levels by increasing feed input while reducing capital inputs. Moreover, the maximum output level of HSD and HFC farming households was estimated to reach 9255 kg/ha and 9807 kg/ha, respectively. Results of the production output simulation suggest that the feeding cost per hectare in LSD households should not exceed 25,119 NTD to avoid overfeeding, which may cause culture deterioration and lower survival rates. As such, farmers are advised to adjust their feed costs according to the stocking density to maximize production output. [ABSTRACT FROM AUTHOR]

It has been demonstrated that the sekelsky mothers against decapentaplegic homolog 3 (Smad3) plays an important role in the growth and development of vertebrates. However, little is known about the association between the Smad3 gene and the growth traits of mollusks. In this study, Smad3 from the hard clam Meretrix meretrix (Mm-Smad3) was cloned, characterized, and screened for growth-related single nucleotide polymorphisms (SNPs) in its exons. The full-length cDNA of Mm-Smad3 was 1938 bp, encoding a protein with 428 amino acid residues. The protein sequence included an MH1 (27–135 aa) and MH2 domain (233–404 aa). Promoter analysis showed that the promoter sequence of Mm-Smad3 was 2548 bp, and the binding sites of Pit-1a, Antp, Hb, and other transcription factors are related to the growth and development of hard clams. The phylogenetic tree was divided into two major clusters, including mollusks and vertebrate. The expression level of Mm-Smad3 was predominantly detected in the mantle and foot, while extremely less expression was observed in the digestive gland. The low expression level of Mm-Smad3 was detected at the stages of unfertilized mature eggs, fertilized eggs, four-cell embryos, blastula, gastrulae, trochophore, and D-shaped larvae, whereas an opposite trend was observed regarding the highest expression at the umbo larvae stage (p < 0.05). In the mantle repair experiment, the time-course expression profiles showed that compared to the expression level at 0 h, Mm-Smad3 significantly decreased at 6 h (p < 0.05) but increased at 12 and 48 h. Further, the association analysis identified 11 SNPs in the exons of Mm-Smad3, of which three loci (c.597 C > T, c.660 C > T, c.792 A > T) were significantly related to the growth traits of clam (p < 0.05). Overall, our findings indicated that Mm-Smad3 is a growth-related gene and the detected SNP sites provide growth-related markers for molecular marker-assisted breeding of this species. [ABSTRACT FROM AUTHOR]

Paralytic shellfish toxins (PSTs) contamination of seafood has become a growing global problem. However, the molecular response of bivalves, some of the most popular seafoods, to PSP toxins has seldom been reported and the underlying molecular mechanisms of the interactions between Meretrix meretrix bivalves and PSTs-producing dinoflagellates are scarcely known. This study compared the protein expression profiles between PSP toxin-contaminated and non-PSP toxin contaminated M. meretrix, determined proteome responses and identified potential biomarkers based on feeding experiments. Results showed that the content of total PSP toxins in contaminated bivalves was 40.63 ± 4.08 μg saxitoxin (STX) equivalents per gram, with 95.3% in hepatopancreas, followed by gill (1.82%) and foot (1.79%). According to two-dimensional gel electrophoresis (2-DE), 15 differentially expressed proteins (at least 2-fold difference) between the hepatopancreas of bivalves with and without PSP toxins were detected. Eight of them were successfully identified by MALDI-TOF MS. These were catalase, protein ultraspiracle homolog, G2 and S phase-expression protein, paramyosin, Mn-superoxide dismutase, response regulator receiver domain-containing protein, sarcoplasmic calcium-binding protein and major facilitator superfamily transporters. The differences in the expression levels of the last three proteins involving in cell signaling, structure and membrane transport were 4.2, 5.3 and 4.9-fold, respectively. These proteins could be further developed as potential biomarkers. The other two up-regulated proteins, Mn-superoxide dismutase and catalase, were involved in cell defence mechanisms against oxidative stress, suggesting PSP toxin acts as xenobiotics and poses oxidative stress in bivalves. This study gives insights into the response of bivalves to PSP toxin-producing dinoflagellate at the proteomic level and the potential of using 2-DE to develop specific protein markers in bivalves. [ABSTRACT FROM AUTHOR]

The hard clam Meretrix meretrix, which has been traditionally used as medicine and seafood, was used in this study to isolate antioxidant peptides. First, a peptide-rich extract was tested for its protective effect against paraquat-induced oxidative stress using the nematode model Caenorhabditis elegans. Then, three novel antioxidant peptides; MmP4 (LSDRLEETGGASS), MmP11 (KEGCREPETEKGHR) and MmP19 (IVTNWDDMEK), were identified and were found to increase the resistance of nematodes against paraquat. Circular dichroism spectroscopy revealed that MmP4 was predominantly in beta-sheet conformation, while MmP11 and MmP19 were primarily in random coil conformation. Using transgenic nematode models, the peptides were shown to promote nuclear translocation of the DAF-16/FOXO transcription factor, a pivotal regulator of stress response and lifespan, and induce the expression of superoxide dismutase 3 (SOD-3), an antioxidant enzyme. Analysis of DAF-16 target genes by real-time PCR reveals that sod-3 was up-regulated by MmP4, MmP11 and MmP19 while ctl-1 and ctl-2 were also up-regulated by MmP4. Further examination of daf-16 using RNA interference suggests that the peptide-increased resistance of C. elegans to oxidative stress was DAF-16 dependent. Taken together, these data demonstrate the antioxidant activity of M. meretrix peptides, which are associated with activation of the stress response factor DAF-16 and regulation of the antioxidant enzyme genes. [ABSTRACT FROM AUTHOR]