Persistent organic pollutant residues in the sediments and mollusks from the Bohai Sea coastal areas, North China: An overview

The Bohai Sea costal area is one of the most developed zones of China and the sewage water from populous and developed cities, including Beijing, Tianjin. Qinhuangdao and Dalian is discharged into the Bohai Sea. Additionally, its semi-enclosed characteristic restricts water exchange, which leads to high accumulation of pollutants in the environment. This overview presents the residues of 6 classes of Persistent Organic Pollutants (POPs). including PAHs, DDTs. HCHs, PCBs and PCDD/Fs. in the sediments and mollusks of the Bohai Sea through analyzing previous literatures. In the sediments. the highest PAH concentrations were detected in the vicinities of Qinhuangdao, while the northeast corner of the Bohai Bay possessed the highest levels of DDTs and PCBs. The investigations on HCHs and PCDD/Fs distributions on the whole sea scale have not been reported. In mollusks, PAH concentrations were in the same order of magnitude in the whole Bohai Sea, so were DDTs. HCHs and PCBs, while the outlier maximum values of PCDDs and PCDFs occurred in Yingkou. In general, the POPs residues in mollusks collected from Shandong Province were higher than the other areas. The compositions of DDTs, HCHs and PCBs in sediments indicated their recent usage. By comparing POP concentrations in sediments with the recommended criterions, it was shown that some individual PAH compounds occasionally associated with adverse biological effects in the vicinities of the Liaodong Bay and Qinhuangdao, and the Liaohe River Estuary were heavily contaminated with DDTs, but PCBs were all below the thresholds. In order to reveal the transference and transformation of POPs in the environment, further studies concerning with their behavior, fate and bioaccumulation in the different trophic levels should be programmed. Moreover, laws and regulations should be enforced to ban the illegal usage of POPs-containing pesticides to guarantee health of the environment and human. (C) 2008 Elsevier Ltd. All rights reserved.

Response of macrozoobenthos communities to ecological engineering remediation in a hypertrophic urban lake

The relationship of macrozoobenthos communities with some environmental variables, and their response to the ongoing restoration measures were studied in a small hypertrophic urban lake near the Yangzte River, China. Twenty taxa including 9 oligochaetes, 7 insects, 2 mollusks and two other animals were found during March 2005 to May 2006. The reappearance of some indigenous macrozoobenthos species showed that the ecological engineering remediation carried out was helpful for the recovery of the macrozoobenthos communities. Through canonical correspondence analysis (CCA), it was detected that temperature (T), conductivity (COND), Secchi depth/deep (SD/Deep), TN, total suspended solids (SS) and chemical oxygen demand (CODcr) were significant environmental factors that influenced the pattern of macrozoobenthos. Limnodrilus hoffmeisteri, Tanypus sp. and Alocinma longicornis could be used as potential bio-indicators in monitoring the development of ecological restoration.

Macrozoobenthic community of Poyang Lake, the largest freshwater lake of China, in the Yangtze floodplain

Poyang Lake (Poyang Hu) is located at the junction of the middle and lower reaches of the Yangtze (Changjiang) River, covering an area of 3283 km(2). As one of the few lakes that are still freely connected with the river, it plays an important role in the maintenance of the unique biota of the Yangtze floodplain ecosystem. To promote the conservation of Poyang Lake, an investigation of the macrobenthos in the lake itself and adjoining Yangtze mainstream was conducted in 1997-1999. Altogether 58 benthic taxa, including, 22 annelids, 8 mollusks, 26 arthropods, and 2 miscellaneous animals, were identified from quantitative samples. The benthic fauna shows a high diversity and a marine affinity. The standing crops of benthos in the lake were much higher than those in the river, being 659 individuals/m(2) and 187.3g/m(2) (wet mass) in the main lake, and 549 individuals/m(2) and 116.6 g/m(2) in the lake outlet, but only 129 individuals/m(2) and 0.4g/m(2) in the river. The dominant roup in the lake was Mollusca, comprising 63.4% of the total in density and 99.5% in biomass. An analysis of the functional feeding structure indicated that collector-filterers and scrapers were predominant in the lake, up to 42.2% and 24.7% in density and 70.2% and 29.2% in biomass, respectively, while shredders and collector-gatherers were relatively common in the river. The present study was restricted to the northern outlet and the northeast part of Poyang Lake. A scrutiny is required for the remaining areas.

Microcystin accumulation in freshwater bivalves from Lake Taihu, China, and the potential risk to human consumption

The potential risk through ingestion of microcystins (MC) in contaminated mollusks has not been well studied. The present paper studied seasonal changes of MC content (determined by liquid chromatography-mass spectrometry) in various organs of three species of bivalves (Cristaria plicata, Hyriopsis cumingii, and Lamprotula leai) in Lake Taihu, China, where toxic cyanobacterial blooms occurred. Coinciding with peaks of seston MC (maximum, 5.7 mu g/L) and MC in cyanobacterial blooms (maximum, 0.534 mg/g), most organs showed sharp MC peaks during the summer, indicating both fast uptake and fast depuration by bivalves. Because hepatopancreas and intestine had considerably higher MC content than other organs, they are the most dangerous for human consumption. Both the present and previous studies show that the hepatopancreatic MC and total tissue MC often are correlated in various aquatic invertebrates. During the peak of the cyanobacterial blooms, C. plicata had higher hepatopancreatic MC content than the other bivalves, whereas H. cumingii had higher intestinal MC content than the other bivalves. Estimated daily intakes for humans from the consumption of whole tissues of the three bivalves were 0.48 to 0.94 mu g MC-LR equivalent/kg body weight (12- to 23.5-fold the tolerable daily intake value proposed by the World Health Organization), which indicates a high risk for humans consuming these bivalves.

Influence of environmental variables on macroinvertebrates in a macrophyte-dominated Chinese lake, with emphasis on the relationships between macrophyte heterogeneity and macroinvertebrate patterns

The benthic community structure in Baoan Lake was examined in relation to lake water physicochemical characteristics and biological parameters. Seventy macroinvertebrate taxa were identified, and mollusks constituted the dominant group and accounted for more than 80% of the total abundance. Assemblages were composed mainly of scrapers (81.5%) and collector-gatherers (roughly 10%). Three plant variables (richness, total cover, and total biomass) were strongly correlated with the faunal gradient (p<0.05). Other predicator variables were Cl-, SiO2, and chemical oxygen demand. Because of the importance of macrophytes in structuring benthic assemblage in this lentic system, the spatial heterogeneity of macrophytes also influenced the pattern of macroinvertebrates. Seven lake regions were uniquely characterized according to primary macrophyte composition and biomass. There were significant differences for macroinvertebrate taxa richness, abundance, and biodiversity among the seven macrophyte regions.

Structure of macrozoobenthos of the East Dongting nature reserve, with emphasis on relationships with environmental variables

Ecological survey of macrozoobenthos assemblages was carried out at 32 sites in the East Dongting Nature Reserve, located in the northern region of the East Dongting Lake in the middle basin of the Yangtze River, China. All total 51 taxa including 18 oligochaetes, 15 mollusks, 14 insects and four other animals were recorded. Mollusks composed the dominant group and accounted for more than 70% of the total abundance. Assemblages were composed mainly of scrapers (66.7%) and collector-gatherers (nearly 20%), and to a lesser extent collector-filterers (roughly 12%), predators (ca. 7%), and shredders (ca. 6%). Two-way indicator species analysis, detrended correspondence, and canonical correspondence analysis (CCA) were employed to identify the relationships between macrozoobenthos assemblages and environmental variables. Thirty-two sites were separated into four site groups based on composition and relative abundance of benthic macroinvertebrates. CCA detected that water depth, pH, conductivity, SiO2, total nitrogen, total phosphorus, alkalinity, hardness, and Ca2+, were significant environmental factors influencing the pattern of macozoobenthos. In this minimal subset, water depth, pH, alkalinity and hardness were the most influential variables.

Cloning, characterization and expression of ferritin subunit from clam Meretrix meretrix in different larval stages

Shell formation is one of the important events during larval development and metamorphosis in bivalves. However, the molecular mechanisms and environmental cues regulating shell initiation and growth are unclear. Here, we report that ferritin, a principal protein for biological iron storage and metabolism, might play a role in larval shell development of the bivalve mollusk Meretrix meretrix. A full-length ferritin subunit cDNA, named as MmeFer, was cloned and characterized. The MmeFer mRNA expression in different developmental stages, from trochophore to post larvae, was analyzed by real-time reverse transcription polymerase chain reaction (RT-PCR). MmeFer mRNA expression in larvae of later developmental stages increased at least 8-fold following trochophores. Moreover, the temporal and spatial expressions of MmeFer mRNA were examined by whole mount in situ hybridization. In the trochophore stage, MmeFer was detectable where it was supposed to be for shell initiation. In the later developmental stages, MmeFer was found near digestive glands and mantle that secret larval shell. MmeFer expression was also detected in larvae cultured in artificial seawater with different iron concentrations ranging from 0 to 100 mu M. These results suggest that ferritin may play a role in the shell formation of mollusks. (C) 2009 Elsevier Inc. All rights reserved.

A novel C1q-domain-containing protein from Zhikong scallop Chlamys farreri with lipopolysaccharide binding activity

The C1q-domain-containing (C1qDC) proteins are a family of proteins characterized by a globular C1q (gC1q) domain in their C-terminus. They are involved in various processes of vertebrates and supposed to be an important pattern recognition receptor in innate immunity of invertebrates. In this study, a novel member of C1q-domain-containing protein family was identified from Zhikong scallop Chlamys farreri (designated as CfC1qDC) by expressed sequence tag (EST) and rapid amplification of cDNA ends (RACE) approaches. The full-length cDNA of CfC1qDC was of 777 bp, consisting of a T-terminal untranslated region (UTR) of 62 bp and a 3' UTR of 178 bp with a polyadenylation signal sequence AATAAA and a poly (A) tail. The CfC1qDC cDNA encoded a polypeptide of 178 amino acids, including a signal peptide and a C1q-domain of 158 amino acids with the theoretical isoelectric point of 5.19 and the predicted molecular weight of 17.2 kDa. The C1q-domain in CfC1qDC exhibited homology with those in sialic acid binding lectin from mollusks and C1qDC proteins from higher vertebrates. The typical 10 beta-strand jelly-roll folding topology structure of C1q-domain and the residues essential for effective packing of the hydrophobic core were well conserved in CfC1qDC. By fluorescent quantitative real-time PCR, mRNA transcripts of CfC1qDC were mainly detected in kidney, mantle, adductor muscle and gill, and also marginally detectable in hemocytes. In the bacterial challenge experiment, after the scallops were challenged by Listonella anguillarum, there was a significant up-regulation in the relative expression level of CfC1qDC and at 6 h post-injection, the mRNA expression reached the maximum level and was 4.55-fold higher than that of control scallops. Similarly, the expression of CfC1qDC mRNA in mixed primary cultures of hemocytes stimulated by lipopolysaccharides (LPS) was up-regulated and reached the maximum level at 6 h post-stimulation, and then dropped back to the original level gradually. In order to investigate its function, the cDNA fragment encoding the mature peptide of CfC1qDC was recombined and expressed in Escherichia coli BL21 (DE3). The recombinant CfC1qDC protein displayed a significantly strong activity to bind LIDS from E. coli, although no obvious antibacterial or agglutinating activity toward Gram-negative bacteria E. coli JM109, L. anguillarum and Gram-positive bacteria Micrococcus luteus was observed. These results suggested that CfC1qDC was absolutely a novel member of the C1qDC protein family and was involved in the recognition of invading microorganisms probably as a pattern recognition molecule in mollusk. (c) 2008 Elsevier Ltd. All rights reserved.

Construction and characterization of two bacterial artificial chromosome libraries of Zhikong scallop, Chlamys farreri Jones et Preston, and identification of BAC clones containing the genes involved in its innate immune system

Large-insert bacterial artificial chromosome (BAC) libraries are necessary for advanced genetics and genomics research. To facilitate gene cloning and characterization, genome analysis, and physical mapping of scallop, two BAC libraries were constructed from nuclear DNA of Zhikong scallop, Chlamys farreri Jones et Preston. The libraries were constructed in the BamHI and MboI sites of the vector pECBAC1, respectively. The BamHI library consists of 73,728 clones, and approximately 99% of the clones contain scallop nuclear DNA inserts with an average size of 110 kb, covering 8.0x haploid genome equivalents. Similarly, the MboI library consists of 7680 clones, with an average insert of 145 kb and no insert-empty clones, thus providing a genome coverage of 1.1x. The combined libraries collectively contain a total of 81,408 BAC clones arrayed in 212 384-well microtiter plates, representing 9.1x haploid genome equivalents and having a probability of greater than 99% of discovering at least one positive clone with a single-copy sequence. High-density clone filters prepared from a subset of the two libraries were screened with nine pairs of Overgos designed from the cDNA or DNA sequences of six genes involved in the innate immune system of mollusks. Positive clones were identified for every gene, with an average of 5.3 BAC clones per gene probe. These results suggest that the two scallop BAC libraries provide useful tools for gene cloning, genome physical mapping, and large-scale sequencing in the species.

CfLGBP, a pattern recognition receptor in Chlamys farreri involved in the immune response against various bacteria

Lipopolysaccharide and beta-1, 3-glucan binding protein (LGBP) is a kind of pattern recognition receptor, which can recognize and bind LPS and beta-1, 3-glucan, and plays curial roles in the innate immune defense against Gram-negative bacteria and fungi. In this study, the functions of LGBP from Zhikong scallop Chlamys farreri performed in innate immunity were analyzed. Firstly, the mRNA expression of CfLGBP in hemocytes toward three typical PAMPS stimulation was examined by realtime PCR. It was up-regulated extremely (P < 0.01) post stimulation of LPS and beta-glucan, and also exhibited a moderate up-regulation (P < 0.01) after PGN injection. Further PAMPs binding assay with the polyclonal antibody specific for CfLGBP proved that the recombinant CfLGBP (designated as rCfLGBP) could bind not only LPS and beta-glucan, but also PGN in vitro. More importantly, rCfLGBP exhibited obvious agglutination activity towards Gram-negative bacteria Escherichia coil, Gram-positive bacteria Bacillus subtilis and fungi Pichia pastoris. Taking the results of immunofluorescence assay into account, which displayed CfLGBP was expressed specifically in the immune cells (hemocytes) and vulnerable organ (gill and mantle), we believed that LGBP in C farreri, serving as a multi-functional PRR, not only involved in the immune response against Gram-negative and fungi as LGBP in other invertebrates, but also played significant role in the event of anti-Gram-positive bacteria infection. As the first functional research of LGBP in mollusks, our study provided new implication into the innate immune defense mechanisms of C. farreri and mollusks. (C) 2010 Elsevier Ltd. All rights reserved.

Physiological and immune responses of zhikong scallop Chlamys farreri to the acute viral necrobiotic virus infection

The Zhikong Scallop, Chlamys farreri, is one of the most Important bivalve mollusks cultured in northern China However, mass mortality of the cultured C farreri has posed a serious threat to the maricultural Industry in recent years. Acute Viral Necrobiotic Virus (AVNV) is believed as an important etiological agent causing the scallop mass mortalities To understand the mechanism behind the AVNV associated scallop disease and mortality, we assessed the physiological and immune responses of C farreri to the virus infection using oxygen consumption rate, ammonium-nitrogen excretion rate, hemocyte copper, zinc superoxide dismutase gene expression, and plasma superoxide dismutase activity and alkaline phosphatase activity as indicators Scallops challenged by AVNV at 25 C developed typical disease signs 2 days after virus injection Before the disease manifested, scallop oxygen consumption and NH4+-N excretion rates rose and then fell back. Real-time PCR revealed that the hemocyte cytosol Cu, Zn SOD gene expression was upregulated followed by recovery The plasma SOD activity, however, augmented consistently following virus injection Moreover, plasma AKP activity first lowered and then elevated gradually to the highest level at 24 h post virus injection Scallops challenged by AVNV at 17 degrees C neither developed notable disease nor showed obvious responses that could be associated with the virus infection. While the results suggested a correlation between the elevated seawater temperature and the AVNV infection associated C farreri mortalities, they also indicated that the viral infection provoked multiple physiological and immune responses in the host scallops (C) 2010 Elsevier Ltd All rights reserved

Pharmacological and immunocytochemical investigation of the role of catecholamines on larval metamorphosis by beta-adrenergic-like receptor in the bivalve Meretrix meretrix

Catecholamines regulate several physiological processes in mollusks. Many pharmacological experiments have been conducted to determine the effects of adrenergic agonist and antagonist of catecholamine receptors on Meretrix meretrix metamorphosis. Results showed that adrenaline (AD) and noradrenaline (NA) had substantial effects (p < 0.05) on larval metamorphosis at concentrations ranging from 10 mu M to 100 mu M. 10 mu M beta-adrenergic receptor (AR) agonist isoproterenol showed the same inducement effect as that of NA and AD on metamorphosis, whereas the alpha-AR agonist phenylephrine had no significant effect at concentrations between 0.1 mu M and 100 mu M concentrations (p > 0.05). Furthermore, I mu M beta-AR antagonist propanolol, but not alpha-AR antagonist prazosin, depressed the larval metamorphosis induced by NA or AD. By immunocytochemistry, two cell bodies of beta-adrenergic-like receptor, C/A1, C/A2, were observed in the cerebral/apical ganglion of competent larvae. In addition, there were other immunoreactive dots near C/A1 and C/A2. The results of pharmacology and immunocytochemistry suggests that beta-adrenergic-like receptor located in the larval CNS, might play a considerable role in the larval metamorphosis of M meretrix by AD or NA. (c) 2006 Elsevier B.V. All rights reserved.

Molecular cloning, characterization and expression of a novel serine proteinase inhibitor gene in bay scallops (Argopecten irradians, Lamarck 1819)

Serine protease inhibitors, critical regulators of endogenous proteases, are found in all multicellular organisms and play crucial roles in host physiological and immunological effector mechanisms. The first mollusk serine proteinase inhibitor (designated AISPI) cDNA was obtained from the bay scallop Argopecten irradians by randomly sequencing a whole tissue cDNA library and rapid amplification of cDNA ends (RACE). The full-length cDNA of the scallop serine protease inhibitor was 1020 bp, consisting of a 5'-terminal untranslated region (UTR) of 39 bp, a 3'-terminal UTR of 147 bp with a canonical polyadenylation signal sequence AATAAA and a poly(A) tail, and an open reading frame of 834 bp. The AISPI cDNA encoded a polypeptide of 278 amino acids with a putative signal peptide of 22 amino acids and a mature protein of 256 amino acids. The deduced amino-acid sequence of AISPI contained six tandem and homologous domains similar to that of Kazal-type serine protease inhibitors, including the conserved sequence C-X(7)-C-X(6)-Y-X(3)-C-X(2,3)-C and six cysteine residues responsible for the formation of disulfide bridges, indicating that the AISPI protein from bay scallop should be a member of the Kazal-type serine protease inhibitor family. The temporal expression of AISPI was measured by semi-quantitative RT-PCR after injury or bacterial challenge. After the adductor muscle was wounded or injected with Vibrio anguillarum, the expression of AISPI mRNA in hemolymph was up-regulated and reached the maximum level at 8 and 16 h, respectively, and then progressively dropped back to the original level. The results indicated that AISPI could play an important role in injury healing and immune response in mollusks as it could be induced by injury and bacterial challenge. (c) 2005 Elsevier Ltd. All rights reserved.

Macrobenthic community structure in the Changjiang Estuary and its adjacent waters in summer

On the basis of data collected in the summer of 2006 from 27 sampling stations in the Changjiang Estuary and its adjacent waters, the ecological characteristics of macrobenthos and the relationship between the macrobenthos and the environmental factors were studied using hierarchical cluster and non-metric multidimensional scaling ( MDS). The biomass, abundance, Shannon - Wiener's and Margalef' s indices of the macrobenthos were presented. The results showed that a total of 253 maerobenthic species were found in the research region, and most. of them belong to mollusks and polychaetes. The dominant species were Cossurella dimorpha, Eocylichna cylindrella, Episiphon kiaochowwanense, Nassarius semiplicatus, Ocstergrenia variabilis and Sternaspis scutata. The average abundance of the macrobenthos was (313.15 +/- 233.4) ind. / m(2), and the average biomass was (15.2 +/- 11.2) g/ m(2). The distribution patterns of the abundance and biomass of the macrobenthos were similar. The abundance and biomass in the area close to the estuary were lower than those from the area more distant to the estuary; the central part of the research region had higher abundance and biomass than other parts of the research region. In accordance with the results, four macrobenthic communities with distinct spatial differences were identified. The low abundance and biomass in the area close to the estuary should be caused by the high sedimentation rate. The statistical analysis indicated that the depth is the most important factor affecting the distribution of macrobenthos.

Chromosomal rearrangement in Pectinidae revealed by rRNA loci and implications for bivalve evolution

Karyotype and chromosomal localization of major (18-5.8-28S) and minor (5S) ribosomal RNA genes were studied in two species of Pectinidae, zhikong (Chlamys farreri) and bay (Argopecten irradians irradians) scallops. using fluorescence in situ hybridization (FISH). C. farreri had a haploid number of 19 with a karyotype of 3m + 4sm + 7sm-st + 4st + 1st-t, and A. i. irradians had a haploid number of 16 with a karyotype of 5st + 11t. In C. farreri, the major and minor rRNA genes had one locus each and were mapped to the same chromosome-Chromosome 5. In A. i. irradians, the major rRNA genes had two loci, located on Chromosomes 4 and 8, and the 5S rRNA gene was found at a third chromosome-Chromosome 10. Results of this and other studies indicate that karyotype of A. i. irradians (n = 16, 21 arms) is secondary and derived from an ancestral karyotype similar to that of C. farreri (n = 19, 38 arms) through considerable chromosomal loss and rearrangements. The ability to tolerate significant chromosomal loss suggests that the modal karyotype of Pectinidae and possibly other bivalves with a haploid number of 19 is likely tetraploid; i.e., at least one genome duplication has occurred during the evolution of Bivalvia.