虾夷扇贝C型凝集素母源传递与抑菌作用的初步研究

为研究虾夷扇贝C型凝集素的母源传递及其抑菌作用,实验运用qRT-PCR技术检测了经鳗弧菌刺激后虾夷扇贝卵巢中C型凝集素的表达模式,比较分析了C型凝集素基因在正常虾夷扇贝和鳗弧菌刺激的虾夷扇贝所产的卵及其胚胎发育前期的存在与变化;通过抑菌实验研究了卵胞浆中C型凝集素抑制细菌存活的作用.结果表明,鳗弧菌刺激能够诱导虾夷扇贝卵巢中的C型凝集素mRNA表达量显著变化,最高表达量出现在刺激8h后,为对照组的6.2倍;母体中C型凝集素可以传递给卵和胚胎,刺激组表达量极显著高于正常组,且表达量都随胚胎发育逐渐降低,至受精36 h时分别为正常对照卵的0.3和0.2倍;蛋白终浓度为200和400 μg/mL的卵无细胞体系都具有一定的抑菌作用,与C型凝集素家族抗体反应后,细菌存活率显著上升,说明母源C型凝集素在抑制细菌存活方面发挥了重要的作用.     

Post-molt processes of cuticle formation and calcification in the Japanese mitten crab <Emphasis Type="Italic">Eriocheir japonicus</Emphasis>

This study examines the following in the Japanese mitten crab: (1) the structure of the exoskeleton with special reference to its calcification; (2) the progression of post-molt cuticle formation and calcification. In the crab, the structure and calcification state of the exoskeleton at the molt and during the inter-molt stage were similar to those of other crustaceans. During the inter-molt, the exoskeleton consisted of four cuticle layers; the outermost epicuticle, the exocuticle, the endocuticle and the innermost membrane layer. Intense calcification was observed in the exo- and endocuticle. At the molt, the synthesis of the epi- and exocuticle was already complete, and the addition of the endocuticle began after the molt. Calcification of the exocuticle initiated soon after the molt, but there was a delay between endocuticle matrix synthesis and calcification. Histology showed that the process of calcification was similar to that in other crustaceans. However, calcium concentrations within the exoskeleton continued to increase and never reached the levels of the inter-molt stage at the end of the experiment. This suggests that the Japanese mitten crab is relatively slow to calcify compared to other crustaceans.     

Culture of mantle epithelial cells expressing shell matrix proteins from scallop Patinopecten yessoensis

ABSTRACT:   In molluscs, mantle epithelial cells secrete organic matrix proteins to form shells. In this study, we established a culture of mantle epithelial cells by using the mantle pallial layer of scallops. We aimed to identify the mantle epithelial cells expressing scallop shell matrix proteins and establish a culture system of epithelial cells. After the mantle pallial layer was carefully isolated from the mantle tissue, explant culture was performed at 4°C. Most cells that migrated from the explant tissue were round cells. Most of the adhered cells retained round morphology, while some of the cells adhered to the dish and showed morphology similar to that of epithelial-like and fibroblast-like cells. When the cultured cells were immunostained with a polyclonal antibody against the shell matrix protein, the antibody recognized many of the adhered cells. An estimation of the number of epithelial cells revealed that approximately 70% of the adhered cells were epithelial cells. This is the first report to describe epithelial cells in cultured mantle cells, which express shell matrix proteins. This culture system may be a useful method for characterization of the mantle epithelial cells.     

牙鲆体质量与形态性状的异速生长分析

为分析牙鲆(Paralichthys olivaceus)体质量与形态性状之间异速生长的遗传规律,通过人工受精的方法建立牙鲆19个全同胞家系,测量了牙鲆不同日龄的体质量和形态性状。采用逐步回归方法建立最优联合异速生长模型,将此模型镶嵌到体质量动物模型的每个遗传和环境效应中,进一步分析对多个形态性状异速生长遗传规律。静态异速生长分析表明:体质量与全长之间存在最大异速生长指数(1.415 5),表现为正异速生长,剩余形态性状与体质量之间的异速生长指数为0.061 5~0.718 0,皆表现为负异速生长。全长与尾柄高之间异速生长指数的正遗传相关最大(0.907 8);全长与尾柄长之间的负遗传相关最大(-0.946 8)。不同模型通过统计标准比较,确定模型Ⅰ为进行牙鲆动态异速生长分析的最优随机回归模型。     

牙鲆养殖群体遗传变异的微卫星标记研究

应用10对微卫星引物对中国牙鲆一个养殖群体的30个个体进行了群体遗传结构分析。结果显示，微卫星标记比其他标记具有更高的多态性，10个微卫星座位的等位基因数在4～10之间，有效等位基因数在2．23～5．82之间，平均等位基因数为7．6，群体平均杂合度为0．6960，Hardy-weinberg遗传偏离指数的平均值为0．1774。     

栉孔扇贝×虾夷扇贝单对杂交子一代幼虫ISSR标记的分离方式

通过对栉孔扇贝Chlamys farreri×虾夷扇贝Patinopecten yessoensis正反单对杂交子一代幼虫进行ISSR(Inter Simple Sequence Repeats)标记,分析了双亲位点在杂交子一代中的传递分离方式。从20个ISSR引物中筛选出引物807和834进行扩增,正交家系和反交家系均统计了70条清晰稳定的扩增带。正交家系中双亲和F_1共有位点占总位点数的38.6%,仅由母本或父本传递给F_1的位点分别占总位点数的27.1%和21.4%。反交家系中双亲和F_1共有位点占总位点数的34.3%,仅由母本或父本传递给F_1的位点分别占总位点数的25.7%和22.9%,表明双亲的遗传物质皆传递给了F_1代,证实属间杂交成功。实验结果表明,两家系中F_1均偏向各自的母本。另外,在F_1中还出现了较高比例的非孟德尔分离位点和非亲位点。     

虾夷扇贝高品质脂肪酸闭壳肌家系选育

以虾夷扇贝闭壳肌重量与脂肪酸组成为主要筛选指标,通过对不同选育家系的生长性状进行比较,在筛选出闭壳肌高出柱率家系的同时,以EPA、DHA和ω-3/ω-6脂肪酸比值等重要营养性状作为参考指标进行家系筛选。结果显示,在12个选育家系中,B04、B05、B10家系的出柱率优势最显著;各家系闭壳肌的EPA+DHA占总脂肪酸45%以上,ω-3/ω-6脂肪酸比值范围为10.94~12.69,家系B03、B05、B06、B07、B10、B11与B12在闭壳肌脂肪酸含量上具有更高选育价值;综合分析,判定B05和B10为虾夷扇贝闭壳肌高出柱率且高品质脂肪酸家系。研究表明,虾夷扇贝的生长与闭壳肌的主要脂肪酸组成没有显著相关性,以闭壳肌脂肪酸含量与生长性状指标进行虾夷扇贝复合性状选育是可行的,本研究结果可以为扇贝选择育种工作的深入开展提供科学依据。     

活品流通过程中虾夷扇贝风味品质的变化

为探索采捕后活品贝类品质的变化规律及评价体系,本研究以活品底播虾夷扇贝为研究对象,针对其闭壳肌的感官及理化特性,系统探讨了在活品流通过程中的风味品质变化规律.根据产业现有的流通模式,建立湿运和干运2条模拟流通链进行为期7d的跟踪研究,通过感官及理化分析监测其风味品质的变化情况;感官监测的指标包含气味、滋味、后味、质地及色泽等5个方面,理化分析包括游离氨基酸、核苷酸关联物、糖原、状态指数CI、剪切力以及蒸煮损失率等指标;针对分析检测结果,首先采用主成分分析方法(PCA)分析风味特征的变化规律,再通过偏最小二乘法(PLS2)对感官和理化二者间的相关性进行分析.研究结果发现,流通链初期,采捕后的剧烈胁迫导致风味品质短暂下降,感官分析出现了苦味、酸味等非愉悦描述词;流通链中期,逐渐适应并处于比较稳定的净化环境条件,风味品质得到恢复,风味特征主要体现在甜味、鲜味、海鲜味等愉悦描述词方面;流通链后期,苦味及酸味等非愉悦描述词再次出现.理化指标的分析结果表明,活品流通过程中扇贝的生命状态整体上呈下降趋势.对比干运与湿运,无论是感官品质还是生命状态,均发现干运在较短期运输(≤2d)时优于湿运,湿运则在较长运输期(≥3 d)时显现出一定优势.感官与理化的相关性分析结果表明,二者之间有关联的特征指标分别为,感官特征的甜(T-ST)、鲜(AT-UM)和奶香(O-DR),理化指标的Gly、Glu、Arg、ATP、AEC及CI,这些指标能够较客观地反映采捕后虾夷扇贝活品流通过程中的风味品质差异,可为建立活品贝类流通过程的品质评价体系提供参考.     

Fine structure and differentiation of the alimentary canal in captive-bred Japanese eel Anguilla japonica preleptocephali

ABSTRACT:   The fine structure of the alimentary canal in preleptocephali produced by artificially matured Japanese eel was examined. At 1 day posthatch (dph), the alimentary canal was found only above the dorsal side of the yolk mass, and the epithelium was composed of a single layer of epithelial cells. By 5 dph, the alimentary canal was divided into three segments based on the structure of the epithelial cells: foregut, midgut and hindgut, corresponding to the future esophagus, intestine and rectum, respectively. After 7 dph, the epithelium in the foregut was surrounded by a circular muscle layer, suggesting a role in the transportation of food materials. The epithelial cells of the midgut exhibited well-developed membranous structures, which are deduced to be invaginations of the cytoplasmic membrane. Pinocytotic invaginations and vacuoles were observed in the epithelial cells of the hindgut; this observation suggests that this region is involved in the uptake of food. Significant changes in morphological features of the epithelial cells in each segment were observed until 7 dph; however, these were not evident between 7 dph and 13 dph. Consequently, the differentiation of the alimentary canal was completed by 7 dph, and preleptocephalus had developed the ability to absorb food by 7 dph.     

Epidermal proteases of the Japanese eel

A fluorescent-sensitive assay was used to demonstrate the protease activity in the dorsal skin of Japanese eel (Anguilla japonica). Two distinct extracts were separately prepared from skin mucus and epidermal cell layers, with no mutual contamination. The epidermal extract was sensitive to various substrates, whereas there was no, or only marginal, susceptibility to the same substrates for the mucous extract. Optimum hydrolysis pHs of the epidermal extract was variable and below pH 7.0, and the optimum hydrolysis temperatures were between 40 and 50 °C. In addition, Tos-Phe-Ch₂Cl, chymostatin, CdCl₂, CuCl₂, HgCl₂ and ZnCl₂ inhibited protease activities to different extents. Several other reagents specifically affected the protease activities, and their induced effects were useful for the identification of epidermal proteases. The findings indicate that a proteolytic factor, exhibiting various enzymological specificities, is retained within epidermal cell layers of Japanese eel. This factor is composed of 4 distinct proteases, such as cathepsins L and B-like proteases, a serine protease and an aminopeptidase.