Cryopreservation of strip spawned sperm using non‐programmable freezing technique in the blue mussel Mytilus galloprovincialis

In this study, a non‐programmable freezing technique has been developed with the strip spawned blue mussel (Mytilus galloprovincialis) sperm. The key parameters optimized including (1) cryoprotectant agents (CPAs); (2) cooling temperature; (3) thawing temperature; (4) sugar and amino acid supplementation and (5) sperm to egg ratio. The fertilization rate and/or integrity of plasma membrane and acrosome were used as sperm quality assessment indicators. The highest post‐thaw sperm fertilization rate of 95% was achieved, when sperm were cryopreservated in 8% dimethyl sulfoxide at 7.8 cm above the liquid nitrogen surface and thawed in a 60°C seawater bath. The addition of glucose, sucrose or trehalose in dimethyl sulfoxide did not, whereas 0.8% glycine did significantly improve the post‐thaw sperm fertilization rates. The fluorescent evaluation has demonstrated that the addition of glycine significantly improved the post‐thaw sperm acrosome integrity, revealing a positive role of glycine in the improvement of post‐thaw sperm quality in blue mussels.     

4种天津近海产贝类牛磺酸和糖胺聚糖的粗提及含量比较

以毛蚶(Arco subcrenat)、紫贻贝(Mytilus edulis)、脉红螺(Rapana venosa)和四角蛤蜊(Mactra veneriformis)的软体部分为原料,经水煮法粗提牛磺酸和酶解法粗提糖胺聚糖,并比较了4种贝类中牛磺酸和糖胺聚糖的含量。结果表明,四角蛤蜊、毛蚶、脉红螺中的牛磺酸平均含量分别为2.708、3.532、3.662g/kg,以脉红螺中牛磺酸含量最为丰富;四角蛤蜊、紫贻贝、毛蚶中糖胺聚糖平均含量为8.27%、4.53%、3.87%,以四角蛤蜊的含量最高。     

紫贻贝消化系统的组织学和组织化学研究

对紫贻贝消化系统进行了组织学和组织化学研究。紫贻贝消化系统由消化腺－肝胰遥和消化道－食道、胃（包括晶杆囊）、肠和直肠组成。消化腺为复管腺,腺上皮有消化细胞和嗜碱性细胞两种类型。消化道管壁由粘膜层、粘膜下层和外膜组成,无肌层。粘膜上皮主要为单层柱状纤毛细胞。组织化学研究表明：消化嗜碱性细胞富含蛋白质和ＲＮＡ,消化细胞富含脂类和多种酶类：蛋白酶、非特异性酯酶、脂酶、酸性磷酸酶及碱性磷酸酶。消化道中酶的     

MytiLec,a Mussel R-Type Lectin,Interacts with Surface Glycan Gb3 on Burkitt’s Lymphoma Cells to Trigger Apoptosis through Multiple Pathways

MytiLec; a novel lectin isolated from the Mediterranean mussel (Mytilus galloprovincialis); shows strong binding affinity to globotriose (Gb3: Galα1-4Galβ1-4Glc). MytiLec revealed β-trefoil folding as also found in the ricin B-subunit type (R-type) lectin family, although the amino acid sequences were quite different. Classification of R-type lectin family members therefore needs to be based on conformation as well as on primary structure. MytiLec specifically killed Burkitt''s lymphoma Ramos cells, which express Gb3. Fluorescein-labeling assay revealed that MytiLec was incorporated inside the cells. MytiLec treatment of Ramos cells resulted in activation of both classical MAPK/ extracellular signal-regulated kinase and extracellular signal-regulated kinase (MEK-ERK) and stress-activated (p38 kinase and JNK) Mitogen-activated protein kinases (MAPK) pathways. In the cells, MytiLec treatment triggered expression of tumor necrosis factor (TNF)-α (a ligand of death receptor-dependent apoptosis) and activation of mitochondria-controlling caspase-9 (initiator caspase) and caspase-3 (activator caspase). Experiments using the specific MEK inhibitor U0126 showed that MytiLec-induced phosphorylation of the MEK-ERK pathway up-regulated expression of the cyclin-dependent kinase inhibitor p21, leading to cell cycle arrest and TNF-α production. Activation of caspase-3 by MytiLec appeared to be regulated by multiple different pathways. Our findings, taken together, indicate that the novel R-type lectin MytiLec initiates programmed cell death of Burkitt’s lymphoma cells through multiple pathways (MAPK cascade, death receptor signaling; caspase activation) based on interaction of the lectin with Gb3-containing glycosphingolipid-enriched microdomains on the cell surface.     

硅烷化表面海洋细菌对厚壳贻贝稚贝附着的影响

为探讨厚壳贻贝稚贝对自然微生物膜中海洋细菌的附着行为反应,本论文研究了厚壳贻贝（Mytilus coruscus）稚贝附着与硅烷基化附着基表面、微生物膜密度以及细菌种属系统发育之间的相互关系。研究表明所测海洋细菌均能显著促进厚壳贻贝稚贝的附着;其中Staphylococcus sp. 1和Cobetia sp. 1表现出较低诱导活性,且这两株细菌形成的微生物膜密度与稚贝附着率之间无显著相关性;其他7株海洋细菌均表现出中等程度诱导活性,且所形成的微生物膜密度与稚贝附着率之间呈显著相关性。系统发育分析表明所测海洋细菌对厚壳贻贝稚贝附着的诱导活性与细菌种属无关。因而,硅烷基化表面的海洋附着细菌对厚壳贻贝附着有着显著性促进作用,本研究将为后续开展厚壳贻贝稚贝附着机制奠定了基础。     

MUSSEL CULTIVATION AS A CO-USE IN OFFSHORE WIND FARMS: POTENTIAL AND ECONOMIC FEASIBILITY

More than 50% of the annual worldwide harvest of mussels is produced in Europe. The mussel cultivation in Germany is based on an extensive on-bottom culture and depends entirely on natural resources for food, spat and space. Due to stakeholder conflicts and a lack of spat availability, mussel farmers tend to move offshore where space is not limited and adequate settlement guaranteed. Newcomers – the offshore wind farmers – are covering large areas in the German Bight which in contrast give the opportunity to use these areas in a multifunctional way by accepting mussel cultivation within the wind farms. This study compiles the basic data for offshore mussel cultivation in close vicinity to a designated offshore wind farm in the open sea of the German Bight and employs different case-scenario calculations to illustrate the impact of changing parameter values on overall profitability or non-profitability of this activity. Primary focus is placed on the production of consumer mussels but seed mussel cultivation is also taken into consideration. We show that production of consumer mussels with longline technology is sufficiently profitable even under the assumption of substantial cost increases. This is especially true, if existing capacities could be used. The cultivation of seed mussels depends on the possibility of using existing equipment. A substantial increase of seed mussel prices to at least 0.6 €, given the main cost categories remaining constant, turns this alternative into substantial profitability. This study concludes with providing some recommendations on how favorable terms or actions could further improve profitability of offshore mussel cultivation. Altogether, our results are intended to shed some light on business management topics that future offshore mariculture operators such as traditional mussel farmers should follow in order to be efficient.     

Flash flood simulation and valve behavior of Mytilus galloprovincialis measured with Hall sensors

Mussels close their shell as a protective strategy and the quantification of this behavioral marker may represent an alarm signal when they are exposed to environmental stressors. In the present study, we investigated the ability of the Mediterranean mussel Mytilus galloprovincialis to recover and then the resilience or inertia of valve activity after a pulsing exposition to diverse levels of salinity (5, 10, 20, and 35 PSU as reference value). The trial simulated an event of drastic and sudden reduction of seawater salinity thus mimicking an event of flash flood from intense rain. Valve gaping and movements were measured in continuous cycle for 10 days using a customized magneto-electric device which uses Hall sensors. Results showed that under normal conditions of salinity (35 PSU), the general pattern of valve movements was a continuously open state with sporadic spikes indicating a closing motion. At salinity of 5, PSU mussels reacted by closing their valves, leading to a 77% mortality on the 4th day. At salinity of 10, PSU animals were observed with closed valves for the entire duration of the exposure and no mortality occurred, they showed a significant reduction in the valve activity once the reference value of salinity was re-established. In contrast, salinity of 20 PSU did not trigger a significant behavioral response. Interestingly, there no define rhythms of valve movements were recorded during salinity challenges.     

不同养殖密度对紫贻贝和披针形蜈蚣藻生态混养的影响

通过混养生态系统模拟试验研究了紫贻贝(Grateloupia lanceolata)和披针形蜈蚣藻(Mytilus edulis)不同养殖密度组合下的生态混养状况。选取壳长(45.14±3.85)mm的紫贻贝和长度(62.48±7.38)mm的披针形蜈蚣藻,采用5种湿重配比进行混养实验,分别为G1(1∶0)、G2(1∶0.25)、G3(1∶0.5)、G4(1∶1)和G5(1∶2)。每种组合中紫贻贝密度均为0.63 ind·L~(-1),而披针形蜈蚣藻密度分别为0 g·L~(-1)、1.25 g·L~(-1)、2.5 g·L~(-1)、5 g·L~(-1)、10 g·L~(-1)。36 d后,G3组中紫贻贝特定生长率为(0.21±0.03)%·d-1,极显著高于其它混养组[(0.11±0.028)~(0.21±0.03)%·d-1,P0.01],而单养组的紫贻贝特定生长率仅为(0.063±0.022)%·d-1;G3组中披针形蜈蚣藻特定生长率为(0.96±0.20)%·d-1,极显著高于其它混养组[(0.62±0.16)~(0.96±0.20)%·d-1,P0.01]。G3组生态系统对营养盐(NO_3~--N、NO_2~--N、NH_4~+-N、PO3-4-P)的去除率分别达到(91.38±1.40)%、(96.79±1.97)%、(98.38±2.06)%、(96.86±3.16)%,显著高于G2组(P0.05),而与G4和G5组没有显著性差异(P0.05)。本研究结果表明,当紫贻贝与披针形蜈蚣藻湿重比为1∶0.5时,可以取得较好的生态效应。     

<Emphasis Type="Italic"> Xenostrobus securis </Emphasis>(Lamarck, 1819) (Mollusca: Bivalvia): first report of an introduced species in Galician waters

The presence of the non-indigenous species, the black-pygmy mussel Xenostrobus securis, is reported here for the first time in an intense shellfish farming area off Galicia (NW Spain). Very high concentrations of this mytilid bivalve have colonized estuarine waters located at the inner part of the Ria de Vigo. The invasive role of X. securis is discussed in the context of the wide ecological tolerance of the species and the recent finding of settlements of this species on numerous colonies of the economically-important blue mussel Mytilus galloprovincialis. The mode of introduction of the black-pygmy mussel is also discussed in relation to human management activities.     

The genetic structure of Mytilus chilensis (Hupé 1854) populations along the Chilean coast based on RAPDs analysis

The Chilean blue mussel Mytilus chilensis is an important commercial species. However, little has been published on the population genetics of this species, despite the need to implement management and conservation policies. Randomly amplified polymorphic DNA‐polymerase chain reaction analysis was used to estimate genetic variation within and between eight natural populations along the whole range of its Chilean natural distribution (ca. 1900 km from Arauco (VIII Region) to Punta Arenas (XII Region)). The values of Nei's unbiased genetic distance, D (0.030–0.107), among populations were small, despite the large geographic separation. A mantel test using 50 000 randomizations showed evidence for a significant correlation (r=0.74, P<0.05) between genetic and geographic (coastal) distance. Punta Arenas population was the most genetically differentiated from the others, although the scale of differentiation was not large (D=0.076–0.107). The levels of gene flow (Nm=1.55) found in this study prevent differentiation among populations by genetic drift. This is the result of the long‐lived planktotrophic larvae of M. chilensis, which provides this species with considerable dispersal ability throughout its range, which is favoured by the ocean currents along the Chilean coast. A restricted larval dispersal towards the north due to the Cape Horn Current derived from the West Wind Drift could be the cause of the higher genetic differentiation of Punta Arenas population from the northern populations. For management purposes of the M. chilensis fishery, the results provide no evidence for discrete stocks, with the possible exception of the Punta Arenas population. The present study provides the baseline data in order to continue further characterization of these mussel populations, considering the great increase in aquaculture of this species.