Mantle Tissue Research Articles

Abundance of saccharides and scarcity of glycosaminoglycans in the soft tissue of clam, Meretrix meretrix (Linnaeus)

We investigated presence and distribution of glycosaminoglycans (GAGs) in Meretrix meretrix soft tissue by determining GAG composition in the different parts, namely, mantle edge, foot, gill, adductor muscle, and viscera. The occurrence of glycan ingredients was examined by histochemistry, whereas GAG and general polysaccharide contents in clam tissue were qualified through extraction and determination. Tissue sections stained with alcian blue or periodic acid–Schiff demonstrated the general existence of saccharides and trifling generation of GAGs in clam tissues. GAGs coexisting with glycogens appeared to be primarily produced in the mantle and foot tissues in mucus form by visualization. The GAG content of the polysaccharide extract ranged from 16.8 to 75.8 mg in 10 g of 5 dried tissue materials in comparison with total carbohydrate level in the range of 500–1760 mg, thereby indicating that GAGs were not the major components of polysaccharide extracts. GAG composition only accounted for approximately 4% of total glycan components, which consist of the determinations of amino sugar and uronic acid. The soft tissues of clam contained abundant saccharide compounds but sparse amounts of GAGs. The results will benefit the subsequent development of products made from the polysaccharide components of M. meretrix.

Identification and expression of cathepsin B from the freshwater mussel Cristaria plicata.

Cathepsin B plays crucial roles in host immune defense against pathogen infection. In present study, a cathepsin B gene from the freshwater mussel, Cristaria plicata (CpCathB) was cloned and characterized. The full-length cDNA of CpCathB was 1825 bp, and contained a 5' untranslated region (UTR) of 36 nucleotides, an open reading frame (ORF) of 1044 bp and a 3' UTR of 745 bp with a poly (A) tail. The deduced CpCathB protein was encoded as a preproenzyme with 347 amino acid residues and predicted molecular weight of 38.55 kDa. Sequence alignment revealed that CpCathB protein shared 56% - 60.7% identity comparison with other species. The predicted tertiary structure of CpCathB protein was highly similar to that of human. The CpCathB mRNA was expressed in hemocytes, hepatopancreas, adductor muscle, gills and mantle tissues of healthy mussels, and the highest expression level was in hepatopancreas. The transcripts of CpCathB were increased in hemocytes and hepatopancreas from mussels after Aeromonas hydrophila challenge. Moreover, the recombinant CpCathB was expressed in the Escherichia coli Rosetta-gami (DE3) strain. The maximum titer of the anti-CpCathB polyclonal antibodies was 1:640,000.The CpCathB protein had a higher expression in hepatopancreas and mantle and a lower level in hemocytes.

Non-lethal method for the preparation of metaphase spreads using cultured mantle tissue from live adult abalone.

Metaphase spread preparation in adult abalone has not been successful, which has restricted the applications of karyotyping-based technologies. Here, we present a non-lethal method to enable preparation of metaphase spreads from live adult abalone using a tissue culture method. Mantle tissue fragments from live adult abalone were cultured in vitro and the cultured cells were used for metaphase spread preparation. To retrieve a sufficient number of proliferating cells required for metaphase spread preparation, at least 14 days of culture was required, and culturing the marginal zone of mantle was more optimal than culturing other areas. Additionally, it was shown that simple medium consisting of basal medium, fetal bovine serum and antibiotics could stimulate cellular proliferation followed by metaphase spread preparation.

Nacre formation by epithelial cell cultures from mantle of the black-lip pearl oyster, Pinctada margaritifera.

Mantle tissue from the black-lip pearl oyster, Pinctada margaritifera, was cultured in vitro using sterilized seawater supplemented with 0.1% yeast extract as the culture medium. Granular and agranular epithelial cells, hyalinocytes, and fibroblast-like cells were observed in the initial stages of culture. Epithelial cells later formed pseudopodial cell networks containing clusters of granulated cells, which upon maturation released their colored granules. These granules induced formation of nacre crystal deposits on the bottom of the culture plate. Cultures comprised of only granulated epithelial cells were established through periodic sub-culturing of mantle cells and maintained for over 18mo in a viable condition. Reverse transcriptase PCR of cultured cells demonstrated gene expression of the shell matrix protein, nacrein. To further evaluate the functional ability of cultured granulated epithelial cells, nuclear shell beads were incubated in culture medium containing these cells to induce nacre formation on the beads. Observation of the bead surface under a stereomicroscope at periodic intervals showed the gradual formation of blackish yellow colored nacre deposits. Examination of the bead surface by scanning electron microscopy and energy dispersive X-ray analysis at periodic intervals revealed a distinct brick and mortar formation characteristic of nacre, comprised of aragonite platelets and matrix proteins. Calcium, carbon, and oxygen were the major elements in all stages examined. Our study shows that mantle epithelial cells in culture retain the ability to secrete nacre and can therefore form the basis for future studies on the biomineralization process and its application in development of sustainable pearl culture.

Molecular cloning and characterization of a putative mitogen-activated protein kinase (Erk1/2) gene: Involvement in mantle immunity of Pinctada fucata

Extracellular signal-regulated kinases (ERKs) are conserved and related with protein-serine/threonine kinases that participate in the regulation of multiple biological processes, such as cell survival, cell differentiation, proliferation, metabolism, and inflammation. However, little is known about the roles of this kinase in the pearl oyster. In this study, we cloned and identified an ERK homolog from Pinctada fucata (PfErk). Furthermore, we have unraveled its expressional kinetics after lipopolysaccharide (LPS) and polyinosinic-epolycytidylic acid (poly I:C) immune challenge. Pferk harbored a 5′ untranslated region (UTR) of 12 bp, a coding sequence of 1074 bp, and a 3′ UTR of 882 bp. The putative peptide comprised a predicted molecular mass of 41.19 kDa, with a theoretical pI of 6.15. Sequence analysis showed that it possesses one STK catalytic domain and a conserved His-Arg-Asp (HRD) domain. In addition, a canonical Thr-Glu-Tyr (TEY) dual phosphorylation motif and an ATRW substrate binding site were also identified in the coding protein. Homology assessment of PfErk showed high similarity to Homo sapiens ERK. Phylogenetic analysis supported a close evolutionary relationship with molluscan orthologs. The expression patterns of Pferk were observed in seven different tissues of pearl oyster, with highest expression in the mantle and lowest expression in the digestive gland. Pferk mRNA expression levels were detected at developmental stages, with the highest expression in D-shaped larvae, followed by the 32-cell stage. The mRNA expression of Pferk was upregulated significantly in P. fucata mantle primary cells and mantle tissue after LPS and poly (I:C) treatment, and PfErk phosphorylation levels were activated by LPS and poly (I:C) challenges. Overall, our results suggested that PfErk may play important roles in pearl oyster innate immunity, and provided a new understanding of mantle immunity in the pearl oyster.

Molecular modularity and asymmetry of the molluscan mantle revealed by a gene expression atlas.

BackgroundConchiferan molluscs construct a biocalcified shell that likely supported much of their evolutionary success. However, beyond broad proteomic and transcriptomic surveys of molluscan shells and the shell-forming mantle tissue, little is known of the spatial and ontogenetic regulation of shell fabrication. In addition, most efforts have been focused on species that deposit nacre, which is at odds with the majority of conchiferan species that fabricate shells using a crossed-lamellar microstructure, sensu lato.ResultsBy combining proteomic and transcriptomic sequencing with in situ hybridization we have identified a suite of gene products associated with the production of the crossed-lamellar shell in Lymnaea stagnalis. With this spatial expression data we are able to generate novel hypotheses of how the adult mantle tissue coordinates the deposition of the calcified shell. These hypotheses include functional roles for unusual and otherwise difficult-to-study proteins such as those containing repetitive low-complexity domains. The spatial expression readouts of shell-forming genes also reveal cryptic patterns of asymmetry and modularity in the shell-forming cells of larvae and adult mantle tissue.ConclusionsThis molecular modularity of the shell-forming mantle tissue hints at intimate associations between structure, function, and evolvability and may provide an elegant explanation for the evolutionary success of the second largest phylum among the Metazoa.

Crossing Phenotype Heritability and Candidate Gene Expression in Grafted Black-Lipped Pearl Oyster Pinctada margaritifera, an Animal Chimera.

Grafting mantle tissue of a donor pearl oyster into the gonad of a recipient oyster results in the formation of a chimera, the pearl sac. The phenotypic variations of this chimera are hypothesized to be the result of interactions between the donor and recipient genomes. In this study, the heritability of phenotypic variation and its association with gene expression were investigated for the first time during Pinctada margaritifera pearl production. Genetic variance was evaluated at different levels, 1) before the graft operation (expression in graft tissue), 2) after grafting (pearl sac tissue expression in chimera), and 3) on the product of the graft (pearl phenotype traits) based on controlled biparental crosses and the F1 generation. Donor-related genetic parameter estimates clearly demonstrate heritability for nacre weight and thickness, darkness and color, and surface defects and grade, which signifies a genetic basis in the donor oyster. In graft relative gene expression, the value of heritability was superior to 0.20 in for almost all genes; whereas in pearl sac, heritability estimates were low (h2 < 0.10; except for CALC1 and Aspein). Pearl sac expression seems to be more influenced by residual variance than the graft, which can be explained by environmental effects that influence pearls sac gene expression and act as a recipient additive genetic component. The interactions between donor and recipient are very complex, and further research is required to understand the role of the recipient oysters on pearl phenotypic and gene expression variances.

Seasonal occurrence of Perkinsus spp. and tissue distribution of P. olseni in clam (Soletellina acuta) from coastal waters of Wuchuan County, southern China

Seasonal occurrence of Perkinsus spp. infection in the wild clam, Soletellina acuta, along the coast of Wuchuan County, southern China was investigated over one year using conventional polymerase chain reaction (PCR) method combined with sequencing technique. The tissue distribution of P. olseni in (S. acuta) was also compared among four tissues, namely the gill, mantle, digestive gland, and siphon, using the real-time PCR method. S. acuta populations were concurrently infected by two Perkinsus spp., P. olseni and P. beihaiensis, among different sampling months. Prevalence of Perkinsus spp. infection in the clam ranges from 9.4% to 50.6% and is significantly different among seasons, and was significantly higher in the March sampling period (spring), while it was significantly lower in the June sampling period (summer). Different seasonal patterns of prevalence occur between P. olseni and P. beihaiensis infection in the clam (S. acuta). S. acuta is more prone to be infected by P. olseni than by P. beihaiensis. The results of qPCR assay demonstrated that positive rate of P. olseni in S. acuta differs significantly among the four tissues. P. olseni infected the gill and mantle tissues more preferably than the siphon tissue; however, the concentration of the parasite did not differ significantly among the four tissues.

Bulimulus tenuissimus (mollusca) as a new potential host of Angiostrogylus cantonensis (nematoda), a histological and metabolic study.

The terrestrial gastropod Bulimulus tenuissimus is widespread in South America. It is an intermediate host of many parasites, but there are no records of infection of this snail by Angiostrongylus cantonensis, despite the occurrence of this parasite and angiostrongyliasis cases in the same areas in which B. tenuissimus occurs. For this reason, it is important investigate the susceptibility of B. tenuissimus to A. cantonensis-infection, since it can be used as intermediate host of A. cantonensis, increasing the list of terrestrial gastropods that infect wild and domestic animals and humans with this parasite. The purpose of this study was to evaluate the susceptibility of B. tenuissimus to experimental infection with L1 larvae of A. cantonensis. The snails were exposed to 1200 L1 larvae and it was possible observe many developing larvae in the cephalopedal mass and mantle tissues, with intense hemocyte infiltration and collagen deposition, but no typical granuloma structures were formed. The glucose content and lactate dehydrogenase activity in the hemolymph varied, indicating an increase of anaerobic energy metabolism in the middle of infection, but with a tendency to return to normal values at the end of pre-patent period. This was corroborated by the marked reduction in the glycogen content in the cephalopedal mass and digestive gland in the first and second week after exposure, followed by a slight increase in the third week. The content of pyruvic acid in the hemolymph was 14.84% lower at the end of pre-patent period, and oxalic acid content was 41.14% higher. These results indicate an aerobic to anaerobic transition process. The PAS reaction showed a large amount of glycogen inside the developing larvae and muscular tissues of the cephalopedal mass, indicating that despite the high consumption of this polysaccharide by the parasite, the snail is able to maintain its energy metabolism based on carbohydrates. The results reveal that B. tenuissimus is a robust host, which can live with the developing larvae of A. cantonensis and overcome the metabolic damages resulting from parasitism.

A Rhodopsin-Like Gene May Be Associated With the Light-Sensitivity of Adult Pacific Oyster Crassostrea gigas.

Light-sensitivity is important for mollusc survival, as it plays a vital role in reproduction and predator avoidance. Light-sensitivity has been demonstrated in the adult Pacific oyster Crassostrea gigas, but the genes associated with light-sensitivity remain unclear. In the present study, we designed experiments to identify the genes associated with light-sensitivity in adult oysters. First, we assessed the Pacific oyster genome and identified 368 genes annotated with the terms associated with light-sensitivity. Second, the function of the four rhodopsin-like superfamily member genes was tested by using RNAi. The results showed that the highest level of mRNA expression of the vision-related genes was in the mantle; however, this finding is not true for all oyster genes. Interestingly, we also found four rhodopsin-like superfamily member genes expressed at an very high level in the mantle tissue. In the RNAi experiment, when one of rhodopsin-like superfamily member genes (CGI_1001253) was inhibited, the light-sensitivity capacity of the injected oysters was significantly reduced, suggesting that CGI_10012534 may be associated with light-sensitivity in the adult Pacific oyster.

Hemocytes in the extrapallial space of Pinctada fucata are involved in immunity and biomineralization

In bivalves, the mantle tissue secretes organic matrix and inorganic ions into the extrapallial space (EPS) to form the shells. In addition, more and more evidences indicate the participation of hemocytes in shell mineralization, but no direct evidence has been reported that verifies the presence of hemocytes in the EPS, and their exact roles in biomineralization remain uncertain. Here, we identified hemocytes from the EPS of Pinctada fucata. Numerous components involved in cellular and humoral immunity were identified by proteome analysis, together with several proteins involved in calcium metabolism. The hemocytes exerted active phagocytosis and significantly upregulated the expression of immune genes after immune stimulation. A group of granulocytes were found to contain numerous calcium-rich vesicles and crystals, which serve as a calcium pool. During shell regeneration, some genes involved in calcium metabolism are upregulated. Strikingly, most of the shell matrix proteins were absent in the hemocytes, suggesting that they might not be solely responsible for directing the growth of the shell. Taken together, our results provided comprehensive information about the function of hemocytes in immunity and shell formation.

Spectroscopic imaging: Nuclear magnetic resonance and Raman spectrometry for the detection of collagen cross-linking from giant squid mantle, fin, and tentacle tissues

ABSTRACTThe application of proton nuclear magnetic resonance (1H-NMR) imaging and Raman spectrometry is described for the detection and structural characterization of collagen cross-linking (pyridinoline) in soluble and insoluble collagen fractions obtained by cation-exchange separation of the mantle, fins, and tentacles of giant squid (Dosidicus gigas). The pyridinoline was detected using a fluorometric procedure only in the soluble collagen fractions. However, the pyridinoline was detected using high-performance liquid chromatography (HPLC) only in the insoluble collagen fraction from the tentacles. Meanwhile, in all samples, pyridinoline was detected by 1H-NMR and Raman spectrometry. The higher peak intensities in both methods were observed in the insoluble collagen fraction from the tentacles. The muscles showed a lower decrease in shear force after 30 min of cooking. 1H-NMR and Raman spectrometry provides an assay which is more sensitive than fluorescence and HPLC for pyridinoline detection in squid tissues, suggesting greater potential for the analysis of different types of tissue and collagen fractions.

PFMG1 promotes osteoblast differentiation and prevents osteoporotic bone loss

Nacre is a widely used mineral medicine that has been reported to have beneficial effects in bone remodeling without an increase in inflammation. Water-soluble nacre matrix has been demonstrated to be responsible for the effect, yet core active ingredients are unknown. Pinctada fucata mantle gene 1 (PFMG1) was first discovered in the mantle tissue of Pinctada fucata. The protein has 2 EF-hands, a calcium-binding domain. PFMG1 protein can affect the growth of calcium carbonate crystals in vitro. Here, we demonstrate that PFMG1 affects cell-cycle distribution and promotes preosteoblast proliferation. PFMG1 accelerates preosteoblast differentiation and extracellular matrix mineralization. During the differentiation process, PFMG1 increases the expression level of osteoblastic marker genes and activates the Erk signaling pathway. PFMG1 also accelerates calcium crystal aggregation in culture medium and suppresses osteoclast formation. Moreover, PFMG1 prevents bone loss caused by ovariectomy. RNA sequencing analysis demonstrated that PFMG1 stimulates genes that are associated with tissue development and ossification, which indicated new genes that function in bone remodeling. Our findings demonstrate the therapeutic potential of PFMG1 from nacre as a novel medicine for osteoporosis.-Li, L., Wang, P., Hu, K., Wang, X., Cai, W., Ai, C., Liu, S., Wang, Z. PFMG1 promotes osteoblast differentiation and prevents osteoporotic bone loss.

Sequence features, expression profiles and biochemical characteristics of a sigma class glutathione S-transferase gene (AiGSTσ) from bay scallop Argopecten irradians

Glutathione S-transferases (GSTs) are a class of enzymes that facilitate the detoxification of xenobiotics and also play important roles in innate immunity. In the present study, a novel sigma class GST gene (designated as AiGSTσ) was cloned from the bay scallop Argopecten irradians via rapid amplification of cDNA ends (RACE) technique. The complete cDNA sequence of AiGSTσ consisted of a 5’ untranslated regions (UTR) of 48 bp, a 3’ UTR of 113 bp with a poly A tail and an open reading frame (ORF) of 618 bp. The ORF encoded a polypeptide of 205 amino acid residues with a calculated molecular mass of approximately 23.11 kDa and a theoretical isoelectric point of 5.354. The deduced amino acid sequence of AiGSTσ contained a GST_N domain and a GST_C domain, and exhibited high similarity with other reported sigma class GSTs. In the phylogenetic tree, AiGSTσ was located in the sigma class GSTs sub-branch. The AiGSTσ mRNA transcripts were constitutively expressed in the tissues of hemocytes, muscle, mantle, gill, hepatopancreas and gonad, with the highest expression level in hemocytes, and the mRNA expression levels of AiGSTσ were significantly up-regulated in hemocytes after various pathogen associated molecular patterns (PAMPs) stimulation. The purified recombinant AiGSTσ protein exhibited catalytic activity against the common substrate 1-chloro-2, 4-dinitrobenzene (CDNB) with low thermal stability and narrow optimum pH spectrum. All these results indicated that AiGSTσ was a fragile but efficient antioxidant enzyme and was potentially involved in the innate immune responses of scallop.

Evaluation of different media for cell proliferation in mantle tissue culture of the green mussel, Perna viridis (Linnaeus, 1758)

The aim of the present study was to establish a suitable culture system for tissue explants from the mantle of the green mussel, Perna viridis. The experiments were conducted using healthy, live green mussels in the size range of 75 to 110 g collected from Pulicat Lake, Tamil Nadu. Three different culture media namely M199, Leibovitz L-15 and sterile seawater were used to assess the most suitable medium for growth, proliferation and viability of mantle epithelial cells. The effect of the addition of two supplements viz., 10% foetal calf serum (FCS) and 0.1% yeast extract to the culture media was also evaluated. After carefully isolating the pallial layer from the mantle tissue, 1-2 mm2 size explants were successfully cultured in 12-well plates at 25°C for up to 14 days. Cultures were monitored under light and phase contrast objectives in an inverted microscope. Cell counts were made and cell size was measured for each treatment. Cells were observed to migrate from the periphery of the explant within 24 h after initiation of cultures. The liberated cells were mostly round and were either granulocytes or hyalinocytes. Fibroblast-like cells were also observed. Our results showed that proliferation of epithelial cells from mantle tissue was maximum in seawater medium (7.4x104 cells ml-1), followed by L-15 medium (2.55x104 cells ml-1). Average cell size in seawater medium was 10.72 μm and that in L-15 and M199 media was 8.56 and 6.39 μm, respectively. Adherent cells were also more prominent and higher in number in seawater medium. Supplementation of culture media with 10% FCS and 0.1% yeast extract improved both cell proliferation and cell size in all the three culture media. Four concentrations of 0.1% yeast extract (@ 50 μl, 75 μl, 100 μl, 150 μl ml-1 medium) were tested in the present study and best results were obtained with 100 μl ml-1, with respect to both cell counts and size.

Sensitivity to ocean acidification differs between populations of the Sydney rock oyster: Role of filtration and ion-regulatory capacities

Understanding mechanisms of intraspecific variation in resilience to environmental drivers is key to predict species' adaptive potential. Recent studies show a higher CO2 resilience of Sydney rock oysters selectively bred for increased growth and disease resistance (‘selected oysters’) compared to the wild population. We tested whether the higher resilience of selected oysters correlates with an increased ability to compensate for CO2-induced acid-base disturbances. After 7 weeks of exposure to elevated seawater PCO2 (1100 μatm), wild oysters had a lower extracellular pH (pHe = 7.54 ± 0.02 (control) vs. 7.40 ± 0.03 (elevated PCO2)) and increased hemolymph PCO2 whereas extracellular acid-base status of selected oysters remained unaffected. However, differing pHe values between oyster types were not linked to altered metabolic costs of major ion regulators (Na+/K+-ATPase, H+-ATPase and Na+/H+-exchanger) in gill and mantle tissues. Our findings suggest that selected oysters possess an increased systemic capacity to eliminate metabolic CO2, possibly through higher and energetically more efficient filtration rates and associated gas exchange. Thus, effective filtration and CO2 resilience might be positively correlated traits in oysters.

Rapid and comprehensive discovery of unreported shellfish allergens using large-scale transcriptomic and proteomic resources

[Extract] Discovery of allergens in various food and inhalant sources is central to our understanding of the molecular mechanisms of allergic reactions. Allergen characterization is the most important underlying factor for better patient management and the design and development of novel immunotherapeutics.1 Of the "Big eight" allergen food groups, shellfish presents a unique challenge in terms of allergen discovery due to the large number and diversity of consumed species, leading to heterogeneity of allergen structure and cross-reactivity among various sources. At present, 31 crustacean allergens have been officially registered in the World Health Organization and International Union of Immunological Societies Allergen Nomenclature Database as compared with 4 mollusk allergens due to pitfalls in current allergy discovery approaches. Cosensitization of patients with crustacean and mollusk allergy is often described; however, the current diagnostic approaches to manage these patients are not based on sufficient molecular knowledge of these shellfish allergens.

Feeding of scallop mantle epithelial cell layer causes subacute toxicity against rodents

Japanese scallops Patinopecten yessoensis are one of the main marine products of Hokkaido, Japan. In addition to adductor muscle, scallop mantle tissue is also eaten in Japan. We fed Wistar rats diets containing the scallop mantle epithelial cell layer for 3–8 weeks and compared the outcomes with a control group. No differences in general appearance or behavior were observed between the groups for the first 2 weeks. Thereafter, a significant decrease in food consumption was observed, and the treated rats died between 6 and 8 weeks. Serum biochemistry and histological studies revealed nephrotoxicity and hepatotoxicity in rats fed scallop mantle epithelial cells. Glucose tolerance tests showed that feeding scallop mantle epithelial cells to rats causes a glucose metabolism disorder. A diet containing a water extract from scallop mantle epithelial cells caused toxicity in mice, but a diet containing a methanol extract, which dissolves diarrhetic shellfish poisoning (DSP) toxin, or an extract in heated 0.1 M HCl, which dissolves paralytic shellfish poisoning (PSP) toxin, did not cause toxicity. Mice fed a diet containing pronase-treated water extract neither showed a decrease in their food intake nor death. These results suggest that a water-soluble component other than DSP or PSP toxin causes toxicity.

First characterization report of natural pearl of Pinctada fucata from Gulf of Mannar

The present study is aimed to characterize the natural pearl of Pinctada fucata from Gulf of Mannar by Scanning Electron Microscope (SEM) and Energy Dispersive Studies (EDS). Pearl oysters (P. fucata) from Kayalpattinam, Gulf of Mannar, were landed as a by-catch in the bottom set gill net at a depth of 4m and collected for tissue culture studies. During mantle tissue dissection, a good lustrous, round pearl of 1.5mm size was found in the mantle fold of pearl oyster P. fucata. This evidenced the existence of natural pearl oyster beds and natural pearls in this region. It was analyzed by Scanning Electron Microscopy (SEM) and Energy Dispersive Spectroscopy (EDS) to find out the composition of nacre. Parallel orientation of crystals to form the lamellar formation of nacre is clearly visible in SEM. Pseudo-hexagonal aragonite crystals arranged in a uniform layer and joined together to form a lamella with inter-lamellar matrix. Two forms of calcium (CaO and CaCO3) obtained in EDS analysis. Calcium content in the natural pearl is 66.05% which is clearly reveals the aragonite form. Niobium (Nb) was attained at 6% in natural pearl which is the first report in its kind and it may also play a role in the formation of lustrous layer. There are so many characterization reports available about mother of pearl in shell of different species, but this is the first report of natural pearl from P. fucata.

Enzymatic and antioxidant response of Lamellidens marginalis exposed to mercuric chloride

In recent decades, pollution of freshwater and riverine ecosystems is indorsed to heavy metals contamination. Among various causes, heavy metals are the most concern. Many aquatic organisms have the ability to inhabit in contaminated environments, due to their defense mechanisms that allow detoxification, excretion, anti-oxidant protection and stress response. Lamellidens marginalis have the potential to accumulate the heavy metals. In the present study, the mussels were exposed to 1, 5, 10 and 15 ppm concentration of mercuric chloride for 96 hrs. Enzymatic and antioxidant parameters including catalase (CAT), superoxide dismutase (SOD), lactate dehydrogenase (LDH), hydrogen peroxide (HP) and lipid peroxidation (LPO) levels were used as effective criteria. All the parameters showed responses in gills, abductor muscle, mantle tissue, gills and gonads with exposure to HgCl2. Collectively, these parameters could be used as biomarker for heavy metals contamination in aquatic system using L. marginalis.