外套膜上皮细胞与组织块的离体珍珠囊形成试验

通过对三角帆蚌外套膜上皮组织块培养和上皮细胞培养存包硬核、无包硬核的情况下的钙代谢进行比较分析,用偏光显微镜观察.结果发现,外套膜上皮组织块、上皮细胞在有硬核时与埘照的无硬核培养钙含量无明显差别,外套膜组织块经培养迁移和增值,能形成珍珠囊的上皮细胞,其结缔组织细胞共同围绕珍珠囊上皮细胞组成完整的珍珠囊.偏光显微镜下能观察到舣折射现象,表明产生由碳酸钙结晶形成的片层叠加构成珍珠质.该研究进一步探讨了体外培育珍珠(试管珍珠)的技术,为试管珍珠的研究提供了实验依据.     

三角帆蚌hcSRCR1基因的克隆及在不同壳色选育系中的表达模式

清道夫受体(SR)是一类对化学修饰的脂蛋白具有很强结合活性的糖蛋白家族。本研究通过RACE方法克隆得到三角帆蚌hcSRCR1基因cDNA序列,该序列全长1 000 bp,其中开放阅读框819 bp,编码272个氨基酸,预测分子量为28.16 ku,理论等电点为5.55;预测含有2个SRCR结构域和6个保守的半胱氨酸残基。qRT-PCR和Western blot结果显示,hcSRCR1 mRNA和蛋白表达模式基本相同,均在三角帆蚌外套膜中表达量最高,在其他组织中的表达量普遍较低,且在紫色选育系外套膜组织中的表达量显著高于白色选育系。外套膜原位杂交结果显示,hcSRCR1基因主要在外套膜外褶的内、外上皮细胞层以及腹膜处的上皮细胞层中表达。研究表明,三角帆蚌hcSRCR1基因与贝壳珍珠质颜色形成具有一定相关性,可为进一步研究该基因在珍珠颜色形成过程中的调控机理提供基础资料。     

小片与蚌间的关系对珍珠质量的影响

本研究采用褶纹冠蚌同一个体自体外套膜小片育珠,结果珍珠质量明显提高。采用三角帆蚌制取外套膜小片植入褶纹冠蚌。结果珍珠质量极差,大多数蚌产不出珍珠;采用褶纹冠蚌制取外套膜小片植入三角帆蚌,结果三分之一的蚌能得到质量高于褶纹冠蚌,产量高于三角帆蚌的珍珠。作者认为供片蚌和受片蚌间相适关系影响珍珠质量,研究如何提高褶纹冠蚌小片在三角帆蚌体内成珠的比率,在生产上是很有意义的。     

2种育珠蚌遗传多样性分析

采用ＲＡＰＤ和同工酶凝胶电泳的方法，对三角帆蚌和褶纹冠蚌的遗传多样性进行了分析。结果显示，三角帆蚌群体内的相似度为０．２９１７，褶纹冠蚌的为０．３６３６；两者的群体间相似系数为０．６７２３，遗传距离为０．３２７７。ＥＳＴ和ＬＤＨ同工酶在褶纹冠蚌外套膜及内脏团组织中的表达酶谱均比在三角帆蚌中复杂，２种组织相比则内脏团中的表达酶谱比外套膜复杂。三角帆蚌和褶纹冠蚌同作为亲缘关系比较接近的主要育珠蚌种，两者各有优缺点。在珍珠养殖和育种上，一方面要重视野生资源的保护，避免近亲繁殖；另一方面要建立种质资源库，不断筛选出更多更好的优良性状，提高育珠的质量和产量。     

褶纹冠蚌外套膜外表皮细胞培养研究

以褶纹冠蚌外套膜外表皮上细胞为材料，利用改良培养基础之胎牛血清和其它营养成分，在一定的光照条件下进行细胞培养，实验结果显示：细胞能贴壁生长，从第3代细胞开始培养瓶中有结晶出现，到第4代培养细胞时已有大量结晶形成。光照对该细胞的成功培养和结晶分泌有一定的促进作用。     

2种育珠蚌遗传多样性分析

采用RAPD和同工酶凝胶电泳的方法,对三角帆蚌和褶纹冠蚌的遗传多样性进行了分析.结果显示,三角帆蚌群体内的相似度为0.2917,褶纹冠蚌的为0.3636;两者的群体间相似系数为0.6723,遗传距离为0.3277.EST和LDH同工酶在褶纹冠蚌外套膜及内脏团组织中的表达酶谱均比在三角帆蚌中复杂,2种组织相比则内脏团中的表达酶谱比外套膜复杂.三角帆蚌和褶纹冠蚌同作为亲缘关系比较接近的主要育珠蚌种,两者各有优缺点.在珍珠养殖和育种上,一方面要重视野生资源的保护,避免近亲繁殖;另一方面要建立种质资源库,不断筛选出更多更好的优良性状,提高育珠的质量和产量.     

三角帆蚌珍珠囊形成的研究

本文报道了对三角帆蚌珍珠囊形成过程的研究结果。方法是把石蜡制成的核和一块细胞小片插入蚌的外套膜中，在不同时期取样，进行组织切片。结果表明珍珠囊的形成是细胞小片细胞先形成一层“初生珍珠囊”上皮细胞，由于细胞小片是异体细胞(来源于供片蚌)，因而受到育珠蚌(受体蚌)细胞的“识别”而被排斥，结果初生珍珠囊上皮细胞与基部的细胞脱离，造成死亡并溶解．其后育珠蚌结缔组织最内层细胞再转化为上皮细胞，形成一层“次生珍珠囊”上皮细胞。所以在珍珠囊形成过程中，插入小片细胞和育珠蚌结缔组织细胞在发生一系列形态结构和位置变化的同时，还有细胞“识别”的现象。在水温20℃左右条件下珍珠囊形成约需30天；在尾部插核的蚌，珍珠囊形成比在中部插核的要快；5月手术的珍珠囊形成比10月手术的要快。     

河蚌外套膜的免疫性研究

从组织学角度对褶纹冠蚌的组织切片移植到三角帆蚌以及三角帆蚌同种蚌之间的外套膜移植所产生的免疫反应进行了定性到定量的研究。移植后受体蚌的嗜酸性粒细胞增加，蚕噬细胞中的蚕噬颗粒明显增多，腺细胞则逐渐排空；而小片中的嗜碱性粒细胞增加；同时还报导了三角帆蚌创伤后的伤口愈合原理。     

应用激光共聚焦显微技术研究Ca2+在三角帆蚌组织内的积累与分布

为了探讨淡水贝类Ca2+吸收和转运信号传递机理,采用激光共聚焦技术研究了三角帆蚌不同组织细胞在静息状态下细胞内游离Ca2+浓度,以及水体Ca2+浓度对外套膜组织细胞内钙沉积的影响。试验选取10只健康的三角帆蚌,分别获得外套膜外膜、内膜、斧足、腮及内脏团上表皮组织细胞,短期培养后用Fluo-3/AM荧光探针孵育细胞1 h,观察细胞内Ca2+荧光强度。研究结果表明,蚌不同组织细胞内Ca2+荧光强度存在显著差异,外套膜外膜细胞内Ca2+荧光强度最高,内脏团上表皮细胞的最低(P<0.05);育珠三角帆蚌在相同Ca2+浓度的孵育水平下,外套膜细胞内Ca2+荧光强度比非育珠蚌都有增加的趋势,其中在1.25 mmol/L添加组中,两组外套膜内膜细胞内Ca2+荧光强度差异达到显著水平(P<0.05);不同Ca2+孵育水平对细胞内Ca2+荧光强度有显著影响(P<0.05),随着Ca2+在孵育液中浓度的升高,外套膜细胞内Ca2+荧光强度显著增强(P<0.05),表明外套膜是从外界吸收Ca2+的主要组织,蚌体珍珠的培育增强了其对Ca2+的沉积,并且水体Ca2+浓度在1.25～3.00 mmol/L有助于蚌体内Ca2+的贮藏,这对进...     

三角帆蚌不同蚌龄外套膜细胞的增殖能力及其矿化功能

研究了淡水珍珠贝——三角帆蚌(Hyriopsis cumingii)在不同蚌龄下外套膜组织的细胞增殖及生物矿化相关因子活性。选择5组不同蚌龄三角帆蚌(0.5龄、1龄、2龄、3龄、4龄),通过流式细胞技术分析了外套膜细胞的增殖指数(proliferation index,PrI)和胞内Ca~(2+)浓度,并应用实时荧光定量PCR检测了与生物矿化相关的碳酸酐酶(carbonic anhydrase, CA)、碱性磷酸酶(alkaline phosphatase, ALP)基因的表达并测定其酶活性。结果表明, 2龄蚌的外套膜细胞PrI及胞内Ca~(2+)浓度显著高于其他蚌龄(P0.05);生物矿化相关基因在不同蚌龄的外套膜组织中表达量不同, 1龄三角帆蚌CA基因表达量和CA酶活性最高(P0.05), ALP基因表达量和ALP酶活在0.5龄三角帆蚌中显著高于其他蚌龄(P0.05)。本研究旨为深入探讨三角帆蚌外套膜细胞增殖能力及人工育珠过程中供体蚌的选择奠定基础。     

三角帆蚌瘟病的生态学防治

采用石灰超细微颗粒悬浮法为主控制池塘生态环境，中草药针剂注射为辅的方法，成功的控制了由嵌砂样病毒（Ａｒｅｎａｖｉｒｕｓ）引起的暴发性三角帆蚌瘟病。７０００只育珠蚌两周年成活率９７３％，药物费用仅占产珠成本的二十分之一。结果表明，提高生态环境的ｐＨ并增加其钙浓度可以抑制蚌瘟病。后来应用此法控制鱼、贝类病毒病，推广面积达１２００公倾。     

Freshwater Pearl Culture Technology Development in India

ABSTRACT

Development of science and technology of cultured pearl production in a freshwater environment in India is described. Distribution of Indian pearl mussels, pond mussel Lamellidens marginalis, paddy field mussel L. corrianus, and riverine mussel Parreysia corrugata in relation to environmental variables such as type of soil, nature of sediment substratum, presence or absence of macrophytes (Eichhornia sp., Nechamandra sp., andNymphaea sp.) is described. Food and feeding of the mussels, together with density aspects in culture conditions are discussed. Mussel breeding including specificity in fish hosts, and mussel larval parasitic relationships are discussed. Basic steps involved in indigenous freshwater pearl culture technology are summarized. Pearl culture grafting procedures such as mantle cavity, mantle tissue and gonadal implantation along with different pearl products are also described.     

三角帆蚌珍珠囊细胞的分泌活动

本文对三角帆蚌珍珠囊细胞的分泌活动进行了亚显微结构的研究。发现珍珠囊表皮细胞能积极地进行物质合成 ,这些物质主要是蛋白质、硫酸化粘多糖和中性粘多糖 ,并将这些物质以微绒毛分泌、块状物分泌、细胞间隙分泌和大颗粒分泌等多种方式分泌到珍珠囊腔 ;而位于珍珠囊表皮细胞之间的、来源于外套膜结缔组织的腺细胞则含有丰富的中性粘多糖 ,并通过开口式分泌的方式释放到珍珠囊腔 ,同珍珠囊表皮细胞分泌的物质一起 ,共同参与珍珠结晶层的形成 ;珍珠囊细胞分泌的多样性与珍珠组成的复杂有关 ;珍珠囊细胞的分泌活动具有节律性和区段性的特点 ,这种区段性和节律性的分泌与珍珠多层结晶纤层的形成相适应。     

Growing Giant Clam (Tridacna Derasa) in Aquaculture Effluent

Juvenile giant clams (Hippopus spp. and Tridacna spp.) are highly valuable and popular in the aquarium trade due to their brightly colored mantles with various patterns. Giant clams are unique bivalves in that they possess symbiotic zooxanthellae (Symbiodinium). A previous study by the authors demonstrated the feasibility of culturing giant clams in aquaculture effluent. Among the four species tested (Tridacna derasa, T. gigas, T. maxima, and T. squamosa), T. derasa was the most suitable for culturing in effluent. The present study compared the growth, survivorship, and condition indexes of T. derasa (mean initial shell length about 83mm) cultured in fish culture effluent or seawater for six months. The clams grew significantly faster (1.29 vs. 0.93mm shell length/month) and had marginally significant (p = 0.076) higher survivorship (94.1% vs. 77.7%) than those in control seawater. Total (shell and tissue) and tissue weight indexes (g/mm shell length), and mitotic index (% dividing zooxanthellae) were similar between the treatment and control clams; whereas zooxanthellae density (number of zooxanthellae/g clam tissue) of the clams in the effluent tanks was 2.5 times higher than that in control tanks.     

How cultured pearls acquire their colour

Round nucleated pearls are produced through a surgical operation, where a round nucleus and a mantle tissue ‘saibo’ from donor oyster are inserted into the gonad of the host oyster. The epithelial cells in the mantle tissue proliferate around the nucleus, and thus, the pearl sac is formed. Pearl sac secrets nacre and forms a pearl. The quality and economic value of pearls are assessed by pearl features such as colour, brightness, lustre and shape. Among all these features, colour has been reported as an important economic indicator and has been widely studied by researchers. Generally, pearl colour is affected by the donor oyster which is determined genetically and biological pigments (melanin and carotenoid). Organic matrices, metal ions and other factors have also been reported to influence the colour of a cultured pearl. Recently, multi‐omics methods have been used to study the colour formation of pearl, and some key genes and signal pathways related to the colour formation of pearls have been identified. Nevertheless, the specific mechanism of pearl formation needs further research. The review combines both fresh and sea water pearls focusing on Hyriopsis cumingii and pearl oysters to provide a general overview and understanding for pearl colour formation.     

Donor effect on cultured pearl nacre development and shell matrix gene expression in Pinctada margaritifera reared in different field sites

Understanding the respective roles played by donor and recipient pearl oysters in pearl quality determination in relation to the environment is a challenge for the pearl industry. In most Pinctada species, pearl size is mainly related to recipient oyster growth performance but also relies to some extent on the biomineralisation activity of the pearl sac, a tissue that originates from the donor oyster mantle. We examined donor effect on pearl size in response to culture in the lagoons on Arutua and Apataki atolls. Overall, nacre weight and thickness were greater in Arutua than in Apataki, but sensitivity to the environment differed between donors. Some donors were associated with significantly heavier and thicker nacre in Arutua (I group), while others had similar results at the two sites (NI group). On average, up to 20% of the pearl size could be attributed to the donor but, in group I, donor effect was responsible for up to 36% of nacre weight determination. Additionally, a real‐time PCR expression study of eight matrix protein genes related to biomineralisation in the pearl sac showed that MSI60, pearlin and pif177 were significantly and positively correlated with nacre weight and thickness, with the latter two genes explaining the larger pearl size observed in Arutua. Donor oysters in P. margaritifera therefore play a key role in pearl size improvement, related to the role of the shell matrix protein genes. Understanding such contributions could help in the design of genetic selection plans for specific and adapted donor oyster lines.     

EXPERIMENTAL CULTURE OF THE OCEAN QUAHOG,Arctica islandica

Larvae and early postlarvae of the ocean quahog, Arctica islandica, were reared under experimental hatchery conditions. Mature eggs were stripped from ripe adults and exposed to a dilute solution of ammonium hydroxide for various lengths of time prior to addition of stripped sperm. The larval clams were reared through settlement and metamorphosis using the Wells-Glancy (centrifuged, incubated seawater) method of algal culture and/or modifications of standard hatchery techniques developed by Loosanoff and Davis. Experimental cultures were maintained at various temperatures ranging from 8.5° to 14.5°C. At temperatures of approximately 13°C, the minimum time to settlement was 32 days, while settlement was not observed in a culture maintained between 8.5° and 10.0°C until approximately 55 days after fertilization. Larval growth rates were significantly lower in the culture maintained at 8.5–10.0°C than in cultures maintained at 11.0–14.5°C. An optical micrograph sequence of larval stages from the straight-hinge stage through metamorphosis is presented to facilitate identification of Arctica islandica specimens isolated from plankton samples. While various workers have reported exceedingly low growth rates of juvenile and adult Arctica, growth rates of larval Arctica appear to be fairly “typical” of rates encountered within the class Bivalvia.     

THE CONTROLLED REPRODUCTION OF THE CARPET-SHELL CLAM (Venerupis decussata [L.]): PRELIMINARY RESULTS

Trials to obtain out-of-season spawning in 13 lots of V. decussata are described. When the animals are considered to be ripe they are treated by thermal and chemical stimulus to set off spawning. A preliminary trial clearly indicated the necessity of feeding the animals undergoing conditioning. Spawning with development of larvae was achieved in two instances after conditioning for two months at 20°C. But inconsistent results in other attempts question the reliability of this method. The culture of 2 lots of larvae is described. After 3 months from egg fertilization clams reach 5 mm mean size with 1% survival.     

Low genetic diversity of cultivated spotted hard clam (Meretrix petechialis) in Taiwan

The aquaculture of spotted hard clams, Meretrix petechialis, is a well‐developed industry in Taiwan. Spats have been produced for decades through artificial propagation using broodstocks selected from neighbouring culture farms on the basis of size and maturity, and the produced spats are resold to these culture farms. Although mass mortality occurs frequently in this practice, the effect of inbreeding depression has never been evaluated. Therefore, genetic diversity of 173 spotted hard clams from museums, two culture farms and three purchased populations was examined in this study. Phylogenetic analyses, based on either COI or cyt b fragments, indicated a division into lineages A and B among the samples. Cultured clams, except for one clustered with M. lusoria, museum specimens, and purchased samples from Hsinchu were all grouped into lineage A. The remaining two purchased populations were placed in both lineages. Compared with cultured clams from the Chinese coast in a previous study, our results exhibited a much lower haplotype (0.354 vs. 0.900) and nucleotide diversity (0.00113 vs. 0.00543). However, whether the loss of genetic variation is a consequence of inbreeding or the founder effect in the initial small broodstocks is unclear. The introduction of broodstocks from northern Vietnam and southern China may facilitate the management of genetic diversity. Environmental factors that potentially cause mass mortality require further investigation.