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Cadmium (Cd) is a toxic heavy metal that, when present as a pollutant in the marine environment, is readily accumulated by marine bivalves, causing oxidative stress and tissue damage. This study explored whether short-term depuration could reverse Cd2+-induced toxicity in the ovary of the clam Meretrix meretrix. Clams exposed to 3 mg L-1 Cd2+ for three days showed increased accumulated Cd2+ in their ovaries with obvious tissue damage as shown by loose structure and some apoptotic cells compared with non-exposed clams. Increased oxidative stress in the ovarian tissue was also obvious, as revealed by increased levels of oxidative indicators such as reactive oxygen species (ROS), malondialdehyde (MDA), DNA-protein crosslinking (DPC), and protein carbonylation (PCO) and increased expression levels of genes related to oxidative stress and apoptosis, which included the Bax, Bcl-2, caspase-3, HO-1, Hsp70, NQO1, Nrf2, and MT genes. When the clams were exposed to Cd2+ for three more days, the accumulated Cd2+ level in the ovary increased to more than 10-fold the level in the control clams, accompanied by more severe damage and cell death as well as oxidative stress. However, when the initial three-day Cd2+ exposure was followed by three days of depuration in Cd2+-free seawater, the Cd2+ level in the ovary was reduced by as much as 20%, accompanied by some recovery of tissue damage and reduced oxidative stress, suggesting that short-term depuration may mitigate Cd2+-induced toxicity in M. meretrix, allowing the clams to recover and potentially reducing the risk of Cd2+ exposure from consuming contaminated clams. [ABSTRACT FROM AUTHOR]

Meretrix meretrix is one of the economical shellfish that has been exploited in Kendari Bay, Indonesia. Several studies have been done, but its growth pattern and condition index are not fully known. This study aims to determine its growth pattern and condition index at the Kambu River estuary, Kendari Bay. This research was conducted for 6 months from August 2021 to January 2022. Clams were collected using a rake, cleaned, and brought to the laboratory to be measured for length and weight using calipers and scales with an accuracy of 0.1 mm and 0.01g, respectively. Furthermore, the clam meat was dried using an oven and weighed. Data on growth and condition index were analyzed using simple regression and the Mann-Whitney test, respectively. The results showed that the growth of M. meretrix was in negative allometric conditions, with b values in males and females in the range of 2.20-3.00 and 1.86-2.73, respectively. The condition index of male and female clam ranged from 1.86-2.62 and 1.66-2.41, respectively, and differed significantly by month. The results of the Mann-Whitney test on clam showed that there was no significant difference between August-November and December-January. In conclusion, male and female clam had relatively different patterns based on the observation period, while the condition index was in the skinny and medium category. [ABSTRACT FROM AUTHOR]

In this study, the complete cDNA sequence of argininosuccinate synthetase was cloned in the saltwater clam Meretrix meretrix using the rapid amplification of cDNA ends (RACE) approach. The complete MmASS cDNA measures 1588 bp in length with an open reading frame encoding 415 amino acids. The molecular weight of MmASS was 46.81 kD and the theoretical isoelectric point was 5.51. The deduced MmASS protein comprises six conserved sequences, containing the substrate-binding residues for ATP, aspartate and citrulline. The results of the multiple amino acid alignment and the phylogenetic analysis showed that MmASS exhibited high amino acid sequence identity with other species. MmASS also had a close phylogenetic relationship with Sinonovacula constricta. Overall, similar functions might exist in M. meretrix and other species. Predicted subcellular localization suggested that MmASS is a cytosolic enzyme. The expression levels of MmASS were distributed in all of the examined tissues. MmASS was most highly expressed in the gill (P<0.05), followed by the hepatopancreas. These results indicated that MmASS was involved in various physiological activities of different tissues and may play an important role in the immune mechanisms. [ABSTRACT FROM AUTHOR]

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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]

C-type lectins (CTLs) are a kind of Ca2+-dependent immunoreactive factors, which participated in pathogens recognition and defense. The present study identified a new CTL from hard clam Meretrix meretrix (designated as MmCTL4). The full-length of MmCTL4 cDNA was 608 bp, encoding a presumed signal peptide of 19 bp and a carbohydrate recognition domain (CRD) of 131 bp. The tertiary structure of recombinant MmCTL4 protein (rMmCTL4) was the typical long double-ring structure with three conserved disulfide bonds, and the motifs in Ca2+-binding sites of MmCTL4 were QPN and WSD. The SYBR Green real-time PCR analysis indicated that MmCTL4 was widely expressed in the hemocytes, hepatopancreas and mantle of healthy clams. After Vibrio splendidus stimulation, the temporal expression profile of MmCTL4 mRNA in hemocytes and hepatopancreas increased by 7.8-fold at 6 hpi and 3.9-fold at 12 hpi, respectively. The cDNA fragments encoding MmCTL4 were recombined into pET-32a (+) vectors, and transformed into Escherichia coli BL21 (DE3). The rMmCTL4 with the presence of Ca2+ performed obvious hemagglutination activity, and could agglutinate E. coli , Bacillus subtilis , and Staphylococcus aureus , while it only weakly agglutinate Vibrio parahaemolyticus and fungi P. pastoris. The agglutination activity of rMmCTL4 were significantly inhibited by D-mannose, D-xylose, D-lactose, maltose and lipopolysaccharides. These results indicated that MmCTL4, as a class of typical pattern recognition receptors (PRRs), could protect the host against pathogen invasion in the innate immunity of clams. [Display omitted] • A novel C-type lectin with QPN and WSD motifs was identified in Meretrix meretrix. • MmCTL4 could be strongly induced after Vibrio splendidus stimulation. • The rMmCTL4 exhibited multiple activities in a Ca2+-dependent manner. [ABSTRACT FROM AUTHOR]

Different sources of proteins have various ways for facilitating the gelation process of surimi matrices, leading to the diverse combined effects with transglutaminase (TG). In this study, the effects of two plant proteins, soy protein isolate (SPI) and pea protein isolate (PPI), and two animal non-muscle proteins, egg white protein (EWP) and whey protein isolate (WPI), with TG addition on the gel properties of clam Meretrix meretrix (MM) surimi gels were investigated, and the mechanisms were explored. In the presence of TG, the exogenous protein addition significantly improved (p < 0.05) the gelation process of the MM surimi. The best improvements were obtained with the combination of EWP, where the gel strength, cooking loss, and water holding capacity were enhanced by 113.08%, 67.89%, and 26.30%, respectively. The addition of SPI showed a relatively weak enhancement compared to EWP incorporation, followed by the addition of PPI and WPI. The results of optical microscopy, differential scanning calorimetry, and chemical forces demonstrated that TG preferentially promoted plant proteins to form self-aggregation in the surimi system, and SPI generated more polymers than PPI. In contrast, the interpenetration of animal proteins and MM myofibrillar proteins was improved by TG. EWP formed a double cross-linking network within the MM protein network, while WPI acted as a binder to promote protein-protein interactions. This research could provide novel insights for developing MM resources and establish a theoretical foundation for the use of composite exogenous protein additives to enhance the gel properties of MM surimi products. • Plant/animal proteins could improve the gelation of TG-added MM surimi. • TG generated more polymers between plant proteins within MM gel network. • SPI generated more self-aggregates than PPI in the presence of TG. • The interpenetration of animal proteins with MM surimi was promoted by TG. • TG induced a double cross-linking network between EWP and MM protein. [ABSTRACT FROM AUTHOR]

Meretrix meretrixwas found to be a very abundant waste at Amal Beach, Tarakan City. M. meretrixwaste can reduce heavy metals, especially heavy metals copper (Cu2+), by the adsorption process. Research has been carried out on M.meretrixas an adsorbent of heavy metals copper (Cu2+) using the adsorption method of Shacker through chemical activation using NaOH bases. The SEM resultsshowed that M.meretrixbefore activation had a rough pore surface that was still closed but M. meretrix after activation had a open pore surface that is whyit could be used as an adsorbent. M.meretrixcontains Al (Aluminum), which can be used as an adsorbent. The adsorption results showed that M.meretrixwith activation in the Freundlich isotherm had the highest ability to absorb Cu2+as much as 9,108 mg/g compared to without activation only 0.051 mg/g. It can be concluded that M.meretrixwaste can be used as an absorbent of Cu2+heavy metals.

The effects and mechanisms of four exogenous proteins, soy protein isolate (SPI), pea protein isolate (PPI), egg white protein (EWP), and whey protein isolate (WPI), on the gelation of Meretrix meretrix (MM) surimi were investigated. The exogenous protein addition significantly increased the G* of samples to 1.36-1.60 folds compared to the MM group, and the MM/EWP sample presented a better time-stable structure. Furthermore, the incorporation of SPI and EWP reduced expressible water content by 17.51 % and 21.61 % in MM gel, respectively. Moreover, SPI and PPI interacted with water to act as fillers within the MM matrix, where SPI formed larger self-aggregations than PPI. A double cross-linked network was generated by EWP and MM protein, and WPI increased the interaction with MM protein as a binder. This research could provide a theoretical basis for the processing and utilization of exogenous proteins in MM., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier Ltd. All rights reserved.)

Vibrio as one of the main pathogens of shellfish diseases can cause serious human seafoodborne gastroenteritis and even death. In this study, we analyzed the bacterial communities from the clam, and compared the resistance phenotypes and genotypes of Vibrio spp. from Meretrix meretrix at different growth stages. High-throughput sequencing analysis revealed the predominance of Proteobacteria (50%) in the bacterial community and Vibrio was one of the dominant genera in the clam hepatopancreas in the summer. Vibrio abundance in Meretrix meretrix positively correlated with the water temperature (p

Cadmium (Cd) is one of the most common pollutants in the environment. It can cause irreversible tissue damage and apoptosis in invertebrates. This study investigated the relationship between Cd exposure and oxidative damage and apoptosis in the ovarian cell of the clam Meretrix meretrix. The clams were exposed to different concentrations of Cd2+ (0, 1.5, 3, 6 and 12 mg L−1) for 5 days, and the accumulated level of Cd2+ in the ovarian tissue, and the degree of oxidative damage, changes in morphology and the response of apoptosis-related factors in the ovarian cell were determined. The bioaccumulation of Cd2+ and the levels of reactive oxygen species (ROS), malondialdehyde (MDA), protein carbonylation (PCO), and DNA-protein crosslinking (DPC) in the ovary were found to increase significantly when the clams were exposed to increasing concentrations of Cd2+. The structure of the ovarian tissue was severely damaged, and the ovarian cells displayed an irregular arrangement. The results of AO/EB staining and flow cytometry showed that the apoptotic rate of the ovarian cells increased with increasing Cd2+ concentrations. The activities of caspase-3, -8, -9, and the mRNA levels of p53, Bax and Caspase-3 in the ovary were also significantly increased. Furthermore, the level of p53 mRNA was positively correlated with the levels of MDA, PCO, DPC and ROS, but negatively correlated with the levels of total antioxidant capacity (T-AOC) and Bcl-2 mRNA. Taken together, these results indicated that Cd2+ exposure would result in oxidative damage and apoptosis for the ovarian cells, suggesting that Cd2+ toxicity could negatively affect the reproductive capacity of M. meretrix, thus threatening the reproductive development of the shellfish.

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]

• Post-stress recovery assays separate Meretrix meretrix into proactive and reactive. • Nano-TiO 2 exposure impaired clams' survival and physiological conditions. • High survival rate in proactive clams might change ecosystem population structure. • Proactive clams had better adaptability under nano-TiO 2 exposure. Nano-TiO 2 is inevitably released into aquatic environment with increasing of nanotechnology industries. Study pointed that different individuality showed divergent behavioral and physiological response when facing environmental stress. However, the effects of nano-TiO 2 on tolerance of bivalves with different individualities remain unknown. In the study, clams were divided into two types of individuality - proactive and reactive by post-stress recovery method. It turned out that proactive individuals had quicker shell opening level, stronger burrowing behavior, faster feeding recovery, higher standard metabolic rate and more rapid ammonia excretion ability than reactive individuals after exposed to air. Then, the survival rate, hemocytes response and oxidase activity of classified clams were evaluated after nano-TiO 2 exposure. Results showed that after 30 d exposure, proactive individuals accelerated burrowing behavior with higher survival rate. Moreover, proactive clams had better adaptability and less hemocytes response and oxidative damage than reactive clams. The study highlights the individualities of marine shell fish determine individual capacity to adapt to environmental changes, play important roles in aquaculture and coastal ecosystem health. [ABSTRACT FROM AUTHOR]

Meretrix meretrix was found to be a very abundant waste at Amal Beach, Tarakan City. M. meretrix waste can reduce heavy metals, especially heavy metals copper (Cu2+), by the adsorption process. Research has been carried out on M. meretrix as an adsorbent of heavy metals copper (Cu2+) using the adsorption method of Shacker through chemical activation using NaOH bases. The SEM results showed that M. meretrix before activation had a rough pore surface that was still closed but M. meretrix after activation had a open pore surface that is why it could be used as an adsorbent. M. meretrix contains Al (Aluminum), which can be used as an adsorbent. The adsorption results showed that M. meretrix with activation in the Freundlich isotherm had the highest ability to absorb Cu2+ as much as 9,108 mg/g compared to without activation only 0.051 mg/g. It can be concluded that M. meretrix waste can be used as an absorbent of Cu2+ heavy metals. [ABSTRACT FROM AUTHOR]

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.

Cadmium (Cd) is one of the most important marine environmental pollutants that can cause oxidative damage and apoptosis in living organisms, and mitochondria are the key cell organelles affected by Cd toxicity. In this study, we investigated the effect of Cd on the mitochondria in the gill cells of the clam Meretrix meretrix and the underlying mechanism of mitochondria-mediated apoptosis following exposure to the metal. Exposure of the clams to artificial seawater containing 1.5, 3, 6 and 12 mg L−1 Cd2+ led to swollen mitochondria compared with the untreated clams. The mitochondria also became vacuolated at the higher Cd2+ concentrations. Biochemical assays showed that monoamine oxidase (MAO) activity and mitochondrial membrane potential (Δψm) increased at 1.5 mg L−1 Cd2+, but decreased at higher Cd2+ concentrations, while the activities of malate dehydrogenase (MDH) and cytochrome oxidase (CCO) and the scavenging capacities of anti-superoxide anion (ASA) and anti-hydroxy radical (AHR) all decreased with increasing Cd2+ concentrations. Significant increases in the levels of malondialdehyde (MDA) and H2O2 as well as in the activity levels of caspase-3, -8, and -9 were also observed in the Cd2+-treated clams. The results implied that Cd might induce apoptosis in M. meretrix via the mitochondrial caspase-dependent pathway. [ABSTRACT FROM AUTHOR]

To understand the effect of salinity on the toxicokinetics, oxidative stress, and detoxification of cadmium-exposed Meretrix meretrix , M. meretrix were acclimatized to different salinities (8, 14, 20, 26, and 32 ppt) for 14 d, exposed to 10 μg/L Cd for 7 d, followed by a 28-day depuration period. The internal Cd concentration was determined, and the activities of antioxidant enzymes (superoxide dismutase (SOD), catalase (CAT), and glutathione-S-transferase (GST)), and the malondialdehyde (MDA) content were measured. The mRNA expression levels of antioxidant enzyme (Cu/Zn SOD , CAT) and detoxification-related genes metallothionein (MT) were analyzed. The mean concentrations of Cd in M. meretrix tissues were in the order gill > digestive gland > mantle > axe foot. The Cd uptake rate in the four tissues decreased with increasing salinity (range: 14–26 ppt). The Cd elimination half-lives were the highest at 8 ppt and 14 ppt salinity. Cadmium activated the four oxidative stress-related related enzymes in the gills. At the end of accumulation period, Cd exposure at 20 ppt salinity significantly increased the expression of Cu/Zn SOD. CAT expression was significantly inhibited at 20 ppt salinity, but was induced at 32 ppt. MT mRNA expression was only induced under Cd at 20 ppt salinity. At the end of depuration period, Cu/Zn SOD expression was inhibited at salinities of 8, 14, and 26 ppt. The results indicated that SOD, CAT, GST, MDA, Cu/Zn SOD , CAT , and MT were sensitive to cadmium in a water environment, and can be used as indicators of marine heavy metal pollution. [Display omitted] • The Meretrix meretrix were exposed to 10 μg/L Cd at salinities of 8, 14, 20, 26, and 32. • Cd concentration in tissues was: gill > digestive gland > mantle > axe foot. • Cd uptake rates increased inversely with salinity at the range of 14–26. • The oxidative stress related parameters play important roles in oxidative damage protection. • MT plays an important role in Cd detoxification. [ABSTRACT FROM AUTHOR]

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.

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.

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.

Metallothioneins (MTs) are low-molecular weight cytoplasmic heavy metal binding proteins. MTs can regulate the concentration of essential or non-essential metals in organisms, and have many important biological functions, including detoxification, trace element metabolism, and anti-oxidation. In the present study, we cloned and characterized a metallothionein gene (designated as MmMT) from the hard clam Meretrix meretrix. The complete cDNA sequence of MmMT contained an open reading frame (ORF) of 629 bp, which encoded a protein of 76 amino acids with a predicted molecular mass of 7.66 kDa and a calculated theoretical isoelectric point of 7.24. MmMT is highly similar to previously identified MTs from other species, with typical metallothionein features such as a high cysteine residue content and the absence of histidine and aromatic residues. The mRNA transcripts of MmMT were prevalent in all the tested tissues, and the expression levels of MmMT were highest in the hepatopancreas and hemocytes. During the stimulation of Vibrio splendidus , the mRNA transcripts of MmMT in the hepatopancreas and hemocytes were significantly increased. The Escherichia coli overexpressing MmMT performed strong growth in the media supplemented with CdCl 2 and CuSO 4 compared to the control strains. These results provide useful information for further investigation of the functions of MmMT in metal detoxification and the innate immune system. • A metallothionein gene (MmMT) were highly expressed in hepatopancreas and hemocytes of Meretrix meretrix. • The mRNA expression of MmMT could be significantly induced by Vibrio splendidus stimulation. • Escherichia. coli overexpressing MmMT displayed higher metal tolerance to Cu2+ and Cd2+. [ABSTRACT FROM AUTHOR]

Cathepsin L is widely found in eukaryotes and prokaryotes, and it plays important roles in innate immunity. In the present study, we cloned two cathepsin L genes (designated as MmCTSL1 and MmCTSL2, respectively) from Asiatic hard clam (Meretrix meretrix). The complete sequence of MmCTSL1 cDNA contained a 5′ untranslated region (UTR) of 31 bp, a 3′ UTR of 228 bp with a poly (A) tail, and an open reading frame (ORF) of 1005 bp encoding 334 amino acids with predicted molecular weight of 37.5 kDa and theoretical isoelectric point of 5.27, and contained a signal peptide (from M1 to A16), a protease inhibitor I29 family domain (from W27 to F87), and a papain family cysteine protease domain (from L118 to T333). The complete sequence of MmCTSL2 cDNA contained a 5′ UTR of 50 bp, a 3' UTR of 162 bp with a poly (A) tail, and an ORF of 996 bp encoding a polypeptide of 331 amino acids with predicted molecular weight of 36.8 kDa and theoretical isoelectric point of 7.07. It contained a signal peptide (from M1 to A16), a protease inhibitor I29 family domain (from W30 to F89), and a papain family cysteine protease domain (from L115 to T330). Real-time quantitative PCR analysis demonstrated that MmCTSL1 and MmCTSL2 were widely expressed in all the tested tissues, including adductor muscle, foot, gill, hemocytes, hepatopancreas and mantle, with the highest mRNA expression level in hepatopancreas and hemocytes, respectively. After Vibrio splendidus challenge, the mRNA expression levels of MmCTSL1 and MmCTSL2 in hemocytes and hepatopancreas were both significantly up-regulated with different expression profiles. In hemocytes, the expression levels of MmCTSL1 and MmCTSL2 reached their respective peaks (3.4-fold and 13.0-fold compared with the control, respectively) at 12 h after bacterial challenge, and MmCTSL2 responds earlier than MmCTSL1. In hepatopancreas, the expression levels of MmCTSL1 and MmCTSL2 reached their respective peaks at 6 h (9.0-fold compared with the control) and 24 h (2.8-fold compared with the control) after bacterial challenge, meaning that MmCTSL1 responds earlier than MmCTSL2. At the same time, whether in hepatopancreas or hemocytes, MmCTSL1 persist for a while after the bacterial challenge peak, while MmCTSL2 would quickly return to the initial level after the bacterial challenge peak. These results indicate that cathepsin L may be involved in the immune process of hard clam against V. splendidus with different potential roles. • Two novel cathepsin L genes were cloned in hard clam. • They were highly expressed in hepatopancreas and hemocytes. • Their mRNA transcripts could be significantly induced with different expression profiles post bacteria challenge. [ABSTRACT FROM AUTHOR]

Recycling of massive waste into valuable products promotes two in one advantages of waste recovery and a pollution-free environment. Preparation of Nanohydroxyapatite (nHAp) and its polymer composites using waste sea shells (natural precursor) are effectively useful in making artificial bone implants, bone cement, and toothpaste additives. The present work is focused to obtain nHAp with good crystalline nature and better biocompatibility using Meretrix Meretrix Clam shells as raw materials by the Co-precipitation method. To enhance the physio-chemical properties, polymers were used as capping agents. The functional group, particle size, thermal stability, phase transition, surface morphology, and electronic transition of atoms of the synthesized samples have been analyzed by spectrographic tools such as FT-IR, XRD, TG/DTA, FESEM/EDAX, XPS and HR-TEM with EDAX mapping respectively. Furthermore, the bacterial sensitivity of the sample against bacteria, the efficiency of the sample towards the formation of an apatite layer over the surface and its hemocompatibility nature were studied by antibacterial, SBF analysis and hemolysis assay respectively. This method of synthesis is free from hard chemicals and harmful by-products. Thus, the report of the present study suggests that it is promising to achieve nHAp with enhanced surface modification which indirectly promotes bone-bonding capability with natural living bone. • Synthesis of pure and polymer mediated nanohydroxyapatite for making bone and dental implants using natural precursor. • First to report, Meretrix Meretrix clam shells were used as a precursor to extract nanohydroxyapatite. • This method is free from harmful chemicals such as reactants and toxic residues as a by-product. • Potential to prepare nanohydroxyapatite with a high yield on large scale with a minimum budget. [ABSTRACT FROM AUTHOR]

A quinquennial seasonal study (2015–2019) has been conducted to evaluate the bioaccumulation pattern of trace metals in Meretrix meretrix. The concentration of trace metals in the clam was observed as Cr > Cu > Ni > Zn > Pb > Cd > Hg, (Body> Mantle > Gills), similar to sediments. Contamination Factor of Cu and Cr in sediments showed strong association with the corresponding metal concentration in the body (r = 0.687, r = 0.962), mantle (r = 0.880, r = 0.956) and gills (r = 0.937, r = 0.863). Bioconcentration Factor was high for Cr followed by Ni. Mean Metal Concentration Rate (MMCR) of Cr was high and Hg was low (Body>Mantle>Gills). Our study establishes that the trace metal intake by Meretrix meretrix is associated with seasonal variation, physicochemical factors, sediment texture, chemical speciation and the metabolic stress created within the species induced from increased demand for protein synthesis. The latter resulted in the augmented rate of accumulation of Cu and Cr. [Display omitted] • Sediment contamination influenced the bioaccumulation of Cu, Cr, Ni and Zn in Meretrix meretrix & sandy clay sediment functioned as an immediate source for bioavailability of metals. • Visualization of Contamination Factor of sediments with trace metal concentration in body, mantle and gills was achieved by Neural Network Model. • M.M.C.R. is a useful method for calculating the degree of trace metal accumulation per day in sediments and benthic organisms, particularly for long term bioaccumulation studies. • Wet seasons, acidic pH, low salinity & low water temperature favoured metal-specific bioaccumulation in Meretrix meretrix. • Metabolic stress created within the species accelerates feeding activity which in turn intensify the rate of uptake of Cu and Cr. [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]