Development of two novel CYP-antibodies and their use in a PCB exposure experiment with Mytilus edulis

In an attempt to learn more about the cytochrome P450 (CYP) system of mussels, we used protein databases and alignment software to extract highly conserved CYP sequences. From these alignments synthetic peptides were produced and used for rabbit immunisation, which yielded polyclonal antibodies against the CYP families 2 and 4. The antibodies were evaluated with Western Blot and ELISA assays, using digestive gland microsomal samples from the mussel Mytilus edulis. Western Blots revealed immunoreactions for both antibodies. The anti-CYP2 sequence rendered one major immunopositive protein of ≈49 kDa size, and weak signals for proteins of ≈41 and 56 kDa size. The anti-CYP4 sequence rendered two major bands of ≈56 and 59 kDa size, and also a weak immunoreaction with a protein of ≈43 kDa size. ELISA rendered only weak signals even with a 1:50 dilution of IgG-purified serum. A 10-day exposure to Aroclor 1254 did not appear to affect any of the immunopositive proteins, while total PCBs in soft bodies increased from 14–40 ng/g DW in controls to 373–638 ng/g DW in exposed mussels.     

贻贝(Mytilus edulis Linnaeus)唇瓣的扫描电镜观察

本文运用扫描电子显微镜技术研究了贻贝唇瓣的形态结，结果表明，贻贝唇瓣外部形态呈羽毛状，分为光滑面和皱褶面；褶皱面表面被覆浓密纤毛，分有沟嵴和无嵴两部分，界限明显，光滑面纤毛较少，呈簇状，分布不均，在褶皱面中央分界处和光滑面的一侧，有大量球状突起，为杯状细胞所在，纤毛和粘液在食物分拣和运输中起重要作用。     

Fatal attraction: synthetic musk fragrances compromise multixenobiotic defense systems in mussels

We studied interactions of nitromusk compounds musk ketone and musk xylene and polycyclic musks Galaxolide trade mark (HHCB), Celestolide trade mark (ADBI), Tetralide trade mark (AHTN), and Traseolide trade mark (AITI) with multixenobiotic resistance (mxr) transporters in gill tissue of the marine mussel Mytilus californianus (Conrad, 1837). A competitive substrate transport test with rhodamine B was used to assay modulation of transport activity by musks. All tested musks inhibited the transport activity in the low microm range as indicated by increased accumulation of rhodamine B in the tissue. Compared to known substrates of mxr transporters, the effective concentration range was similar to quinidine and about 100 times higher than verapamil. Musk ketone and musk xylene also inhibited efflux of rhodamine B from gill tissue which was loaded with the dye and subsequently incubated with these compounds. Synthetic musk compounds are persistent environmental pollutants in aquatic environments with a high potential to bioaccumulate. As potent inhibitors of mxr transporters they may also play a role as chemosensitizers that enable toxic mxr substrates to accumulate in cells of aquatic organisms.     

A 15-month study of zooplankton ingestion by farmed mussels (Mytilus edulis) in Bantry Bay, Southwest Ireland

There is a growing body of evidence to suggest that bivalve molluscs routinely ingest zooplankton. To elucidate further these observations, a 15-month study of zooplankton ingestion by farmed mussels was conducted using mussel long-lines in Bantry Bay, Ireland. Stomach content analysis of the mussels showed that there was evidence of zooplankton ingestion throughout the sampling period, but that highest mean numbers of zooplankters were ingested by mussels in the spring and summer months. Various zooplankton species were present in mussel stomachs. Harpacticoid copepods were found more often in stomach contents than calanoid copepods, probably due to their proximity to the bivalves' inhalent siphons. Barnacle cyprids featured in large numbers in stomach contents, but only for a period of 3 months which broadly corresponded with their pelagic phase. Sizes of ingested zooplankton ranged from 126 μm to 6 mm, but more of the smaller zooplankters (e.g. crustacean nauplii) were ingested. When lengths of ingested copepods were compared with those found in plankton net samples, it was found that the net-sampled copepods were significantly larger than those found in mussel stomachs, suggesting that mussels select for smaller categories within the zooplankton available to them. Soft bodied zooplankton was rarely found in mussel stomachs but their absence may be due to rapid digestion or they may have been destroyed in the preservation process. Ingestion of zooplankton by bivalves is discussed in the context of the impacts mussel farms have on resident zooplankton populations.     

Feeding Habits of Ocinebrina edwardsi (Mollusca: Prosobranchia) a Common Mussel Drill of the Italian Coasts

Abstract. The shell-boring gastropod Ocinebrina edwardsi (PAYR.) lives associated with Mytilus galloprovincialis LMK on which it preys. Predation is preferentially directed towards small (< 15 mm long) mussels. The predation rate (number of mussels eaten per gastropod per day) for medium sized mussels (16–25 mm long) during a year ranges from 0.03 to 0.11. The predation rate is directly influenced by the sea-water temperature. Predation follows a seasonal cycle reaching a maximum in July and a minimum in January. O. edwardsi drills selectively the median anterior region of the mussel valve, close to the dorsal edge.     

Detection of DNA damage in mussels and sea urchins exposed to crude oil using comet assay

The single-cell microgel electrophoresis assay or the comet assay was used to evaluate DNA damage of dispersed crude oil on sea urchins (Strongylocentrotus droebachiensis) and mussels (Mytilus edulis L.). Sea urchins were exposed to 0.06 and 0.25 mg/L dispersed crude oil in a continuous flow system, while the mussels were exposed to 0.015, 0.06 and 0.25 mg/L dispersed crude oil. Sea urchin coelomocytes and mussel haemocytes were sampled after 4 and 5 weeks exposure, respectively. In the sea urchin coelomocytes, there was a significant concentration-related increase in the percentage of DNA in comet tail. In mussel haemocytes, there was a significantly higher percentage of DNA in comet tail for all treatments compared to the control. The responses were concentration-related up to 0.06 mg/L oil. The two highest exposure concentrations of mussels were not significantly different from each other. These results indicate that the comet assay can be used for biomonitoring of DNA damage in marine invertebrates following oil contamination.     

Toxicity of tributyltin in the marine mollusc Mytilus edulis

Our previous studies have demonstrated that tributyltin (TBT) is genotoxic to the early life stages of marine mussels and worms. Here, the toxicity of TBT to adult organisms was determined using a suite of biomarkers designed to detect cytotoxic, immunotoxic and genotoxic effects. Exposure of adult mussels, Mytilus edulis, to environmentally realistic concentrations of TBTO for 7 days resulted in a statistically significant decrease in cell viability at concentrations of 0.5 μg/l and above. TBT had no effect on phagocytic activity or antioxidant capacity (FRAP assay). There was a statistically significant increase in DNA damage detected using the comet and micronucleus assays between the controls and 0.5, 1 and 5 μg/l of TBTO (P > 0.0005). Furthermore there was a strong correlation between DNA strand breaks (comet assay) and formation of micronuclei (P = 0.0005; R² = 61.5%). Possible mechanisms by which TBT could damage DNA either directly or indirectly are discussed including the possibility that TBT is genotoxic due to its ability to disrupt calcium homeostasis.     

基于F型mtDNA D-Loop的厚壳贻贝(Mytilus coruscus)群体遗传多样性研究

为研究东海区厚壳贻贝(Mytilus coruscus)遗传多样性,以厚壳贻贝F mtDNA D-loop为标记,对浙江省舟山嵊山岛、宁波渔山岛、温州南麂岛、福建省宁德湾和莆田南日岛五个海区的厚壳贻贝群体进行了遗传分析。结果表明,厚壳贻贝各群体的遗传多样性差异不明显,在5个群体中,宁德群体的遗传多样性相对最丰富;将5个群体作为一个整体时,呈现出较高的单倍型多样性和较低的核苷酸多样性。对厚壳贻贝5个群体间的遗传分化系数(Fst)和基因流(Nm)进行检测,结果显示群体间Fst值都很低,但Nm值都很高(Nm绝对值1),表明5个群体间存在丰富的基因交流。但宁德群体与浙江沿海的3个群体(嵊山、渔山、温州)的Fst值相对较高,且差异显著(P0.5),表明宁德群体与这3个群体间出现遗传分化。本研究旨在为海洋经济贝类资源的保护管理提供理论依据。     

贻贝粉酶解液提取复合氨基酸

报道了以脱脂后的贻贝粉为原料，采用酶解法从其酶解液中分离提取复合氨基酸的工艺。其收率为２０％，纯度约为９０％。其中必需氨基酸（不计色氨酸）约占复合氨基酸总量的５５％，其氨基酸模式与ＦＡＯ／ＷＨＯ组织确定的模式及人乳模式进行了对照，通过适量添加个别氨基酸可使其配比接近人乳模式。     

贻贝(Mytilus edulis)的核型与减数分裂

利用体内注射ＰＨＡ和秋水仙素的方法，以锶和生殖腺为材料，对贻贝ＭｙｔｉｌｕｓｅｄｕｌｉｓＬｉｎｎａｅｕｓ的核型和减数分裂进行了观察分析，结果为，二倍体数２ｎ＝２８，核型为１０ｍ＋１２ｓｍ＋６ｓｔ，ＮＦ＝５０。没有发现异形性染色体，贻贝的减数分裂与其他动物类似，也没有发现异形染色体的特殊行为。