厚壳贻贝5-羟色胺2A受体(5-HT2AR)基因克隆和时空表达

为探究5-羟色胺2A受体（5-HT 2A receptor， 5-HT2AR）基因在海水贝类生长发育中的作用，本研究通过RACE技术和实时荧光定量PCR（qRT-PCR）克隆了厚壳贻贝5-HT2AR 基因的cDNA全长，并分析该基因的时空表达。结果显示，5-HT2AR基因全长2 636 bp，开放阅读框（ORF）2 124 bp，共编码707个氨基酸。序列分析结果显示该序列与人、小鼠、斑马鱼、长牡蛎和虾夷扇贝等物种分别具有45%、45%、48%、49%和67%的同源性。厚壳贻贝雌雄成体各组织和器官中均有5-HT2AR基因表达，雄性中的鳃表达量最高，而在雌性鳃、外套膜和性腺中的表达稍高于其他组织和器官；推测该基因可能与厚壳贻贝的摄食、对外界环境的感知及促进卵母细胞成熟有关。5-HT2AR基因在厚壳贻贝各发育阶段均有表达，且稚贝的表达量为眼点幼虫的1.4倍，推测5-HT2AR基因可能参与了调控厚壳贻贝幼虫的生长发育过程。本研究为进一步了解5-HT基因家族在双壳贝类中的功能奠定了基础。     

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。     

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.     

Characterization of single‐nucleotide polymorphism markers in the Mediterranean mussel,Mytilus galloprovincialis

The Mediterranean mussel, Mytilus galloprovincialis, is one of the most important aquaculture species in Europe. Appropriate molecular markers are required to evaluate genetic resources and to trace genealogies in breeding programmes for improving mussel culture. Microsatellites have been commonly applied to this purpose in other species. However, Mediterranean mussel microsatellites have demonstrated high frequencies of null alleles that hamper accurate estimates of population parameters and confident parentage inferences. As alternative markers, we have characterized in silico 25 potential single‐nucleotide polymorphism (SNP) markers in the Mediterranean mussel from expressed sequence tag (EST) public databases. The genotyping of SNPs was performed using a single‐base extension approach. Their polymorphism was evaluated in 47 individuals from an Atlantic population. Out of the 25 potential SNPs tested, 12 were technically feasible (producing a single amplicon) and polymorphic. All were biallelic and had an unbiased heterozygosity ranging from 0.160 to 0.504. One SNP was from a mitochondrial gene. The combined potential of nuclear SNPs for parentage assignment gave an exclusion probability of a false couple of parents of 0.9471. These markers will be useful for evaluating resources and tracing genealogies in genetic breeding programmes implemented to solve the main problems of mussel culture.     

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。     

紫贻贝和厚壳贻贝杂交及F1代杂交优势初探

2005年-2007年对紫贻贝和厚壳贻贝种间进行了杂交生产性试验，通过亲贝强化培育、确认雌雄亲贝、改革育苗水体交换方法、投喂混合单细胞藻类、流水培养附着稚贝等技术，获得F₁代。结果表明，各试验组生长特性，正交F₁代﹥紫贻贝﹥厚壳贻贝﹥反交F₁代。其中正交F₁代获附着稚贝3.77亿粒，平均壳长为1.13 mm、平均壳高为0.78 mm，育苗成活率达35.57%。正交F₁除在孵化率上低于紫贻贝、厚壳贻贝外，在壳长、壳高、成活率等方面指标具有一定杂交优势，而反交F1代的杂交优势不明显。     

杂交贻贝同工酶表达的初步分析

运用聚丙烯酰胺凝胶电泳技术,对2种贻贝(厚壳贻贝、紫贻贝)稚贝及通过人工培育所得的二者的杂交后代稚贝同工酶表达状况进行分析.结果显示,所检测5种酶(SOD、MDH、ME、ATP、ADH)在上述3种贻贝中的表达极为相似,MDH在3种贻贝中除r迁移率相同的条带外,在紫贻贝中表达出迁移率较小、表达较弱的条带;SOD和ATP仅在厚壳贻贝中表达出比另外2种多1条迁移率最大的条带,ME则表现为紫贻贝少1条中间的条带,ADH在杂交贻贝和紫贻贝中表达完全相同,厚壳贻贝虽也表现为3条酶带,但迁移率明显不同.试验结果说明,厚壳贻贝和紫贻贝具有较近的亲缘关系,其杂交后代与二者均有一定的相似性,这与传统的分类方法所得的结论相一致.     

低湿度表面的海洋附着细菌对厚壳贻贝附着的影响

从低湿度表面形成自然微生物被膜中分离和纯化出9株海洋附着细菌,并进行细菌16S rDNA基因序列的鉴定以确定其菌名;在18℃、黑暗条件下培养48h后形成单一菌株的微生物被膜,研究了细菌不同初始密度条件下形成微生物被膜的终密度变化及其对厚壳贻贝Mytilus coruscus稚贝附着的影响。同时,将获得的序列与有亲缘关系的序列用Mega软件的CLUSTALW程序进行序列比对和系统发育分析。结果表明:所有测试海洋附着细菌形成微生物被膜的最终密度随初始密度的增加而增加;9株海洋细菌均能不同程度地促进厚壳贻贝稚贝的附着,其中Bacillus sp.2诱导活性最高,其诱导稚贝的附着率为68%;除Pseudoalteromonas sp.3微生物被膜终密度与稚贝附着无显著相关性外(P0.05),其余8株海洋细菌所形成的微生物被膜终密度与稚贝附着均呈显著相关性(P0.05);系统发育分析表明,微生物被膜终密度对稚贝附着的诱导活性与细菌种属无关。研究表明,低湿度表面的海洋附着细菌对厚壳贻贝的附着有明显的促进作用,本研究结果对于今后开展厚壳贻贝稚贝的附着机制研究奠定了基础。     

温度和盐度对厚壳贻贝耗氧率的影响

采用实验生态学方法研究了温度和盐度对厚壳贻贝耗氧率（OR）的影响。根据厚壳贻贝的个体大小分为A、B、C 3组（壳长分别为5.72±0.18、7.12±0.14、8.40±0.19 cm,软体部干重分别为5.75±0.62、10.83±1.23、12.58±2.05 g）,温度设10℃、16℃、22℃、28℃、34℃5个梯度,盐度设14、19、24、29、34共5个梯度。结果表明：（1）温度、个体大小对厚壳贻贝的耗氧率有显著影响（F〉F0.01）,而温度和体重的交互作用对厚壳贻贝的耗氧率无显著影响（FF0.01）。盐度在14~29时,3组厚壳贻贝的耗氧率随盐度的升高而逐渐增加,当盐度升至34时耗氧率下降。厚壳贻贝单位耗氧率（ORS）与软体部干重（W）的关系可用回归方程表示为ORS=a2W-b2,a2值为0.533 9~1.487 3,平均值为0.971 6;b2值为0.307 8~0.566 9,平均值为0.431 7。     

对贻贝养殖中的敌害生物中华豆蟹的繁殖和世代交替研究

本文根据在河北秦皇岛山海关东姜庄贻贝养殖海区三年来的调查研究资料，概述了中华豆蟹对贻贝养殖的危害和繁殖习性，着重探讨了中华豆蟹的世代交替情况，并提出两项改变现有贻贝养殖周期以防止中华豆蟹危害的措施．