Phenotypic indicators for cultured pearl size improvement in the black‐lipped pearl oyster (Pinctada margaritifera): towards selection for the recipient growth performance

The black‐lipped pearl oyster, Pinctada margaritifera, is the most important farmed species in French Polynesia and the basis of the most valuable export industry. Mass production of black pearls relies on a surgical operation requiring tissue from a donor pearl oyster to be grafted, together with a nucleus made of shell, into the gonad of a recipient oyster. Improving pearl size through family selection remains one of the main challenges for future aquaculture development. This study analyses the relative contribution of donor and recipient oysters to pearl size. To this end, hatchery‐produced donor oysters of two batches, large and small (based on shell height), were used to supply grafts for recipients, which were then monitored individually for their growth performance by recording shell height, width, and thickness, and total live weight (flesh + shells) every 6 months (four biometric measurement times) over 20 months of culture. Pearls issued from the two batches of donors showed no significant differences in nacre weight or thickness. In contrast, recipient oyster shell height and total weight were increasingly positively correlated with these pearl size parameters over the culture period, becoming significant at 8 months post‐grafting. Potential therefore exists to use shell height and oyster weight as phenotypic indicators for selective breeding of recipient oysters with high growth performance to increase pearl size in P. margaritifera.     

生长适温下马氏珠母贝黄壳色选系 F1与养殖群体消化酶活力的比较

2006年4月,从流沙港马氏珠母贝养殖群体中挑选11个纯黄壳色个体为繁殖群体建立了马氏珠母贝(Pinctada martensii)黄壳色选系F1(SG1)。同时,随机选取养殖群体中50个个体作为繁殖群体建立了对照组(CG)。2007年10月,从2个组分别选取相同规格的个体,比较了生长适温(15～30℃)下2个组的消化酶活力。结果表明:(1)马氏珠母贝肝胰脏中具有淀粉酶、纤维素酶和蛋白酶活力,在15～30℃条件下,三者活力由高到低依次为淀粉酶、纤维素酶、蛋白酶。(2)在15～30℃范围内,马氏珠母贝淀粉酶活力随温度升高先上升再下降,在温度25℃时达到最大值;SG1组和CG组淀粉酶活力变化范围分别为3.51～5.09μg·min-1·mg-1和2.53～4.04μg·min-1·mg-1;各反应温度下,SG1组淀粉酶活力均高于CG组,在15℃和20℃时,二者差异显著(P0.05)。(3)在15～30℃范围内,马氏珠母贝纤维素酶活力随温度上升而上升,在温度30℃时达到最大值;SG1组和CG组纤维素酶活力变化范围为(2.44～3.22)μg·min-1·mg-1和(2.07～3.12)μg·min-1·mg-1;各实验温度下,SG1组纤维素酶活力均高于CG组,但差异均不显著(P0.05)。(4)在15～30℃范围内,马氏珠母贝蛋白酶活力随温度上升而升高,在温度30℃时达到最大值;SG1组和CG组蛋白酶活力变化范围为(0.075～0.296)μg·min-1·mg-1和(0.067～0.455)μg·min-1·mg-1。在15℃时,SG1组蛋白酶活力大于对照组,差异不显著(P0.05);在20℃、25℃、30℃时,CG组蛋白酶活力显著大于SG1组(P0.05)。本研究结果表明,经过一代壳色选育后黄壳色选系与对照组的消化生理指标存在明显差异,为马氏珠母贝的黄壳色系进一步选育提供依据。     

马氏珠母贝肉酶解产物的抗酒精性肝损伤作用

为探究马氏珠母贝肉酶解产物(Enzymatic hydrolysate from Pinctada martensii,EP)对酒精性肝损伤(Alcoholic liver damage,ALD)的保护作用,该研究将EP超滤分级为截留分子量>10 kD(EP-Ⅰ)、3~10 kD(EP-Ⅱ)和<3 kD(EP-Ⅲ)3个组分,检测其体外抗肝损伤活性及对ALD小鼠肝保护作用的影响。体外试验结果显示,EP-Ⅲ可显著激活体外乙醇脱氢酶(ADH)活性(P<0.01),3个超滤组分均具有一定的体外抗氧化能力且EP-Ⅲ>EP-Ⅱ>EP-Ⅰ;动物试验结果显示,与模型对照组相比,各超滤组分均能够显著降低小鼠血清中谷丙转氨酶(ALT)和谷草转氨酶(AST)活力、小鼠肝脏指数及肝脏中丙二醛(MDA)和甘油三酯(TG)含量,同时显著增强小鼠肝脏中超氧化物歧化酶(SOD)、乙醇脱氢酶(ADH)和乙醛脱氢酶(ALDH)活力,提高肝脏中的谷胱甘肽(GSH)含量。综上,马氏珠母贝肉酶解超滤组分对急性ALD具有一定的辅助保护作用,其中EP-Ⅲ的保护作用效果最佳,其机制可能与加快机体乙醇代谢和减缓乙醇对机体造成的氧化损伤相关。     

马氏珠母贝Bcl-2-like基因克隆及其功能初探

B淋巴细胞瘤-2(B-cell lymphoma, Bcl-2)基因作为一种重要的细胞凋亡调控基因,在内源性细胞凋亡通路中发挥着重要的调控作用。实验利用c DNA末端快速扩增(RACE)技术克隆获得PmBcl-2-like基因cDNA全长序列,并对其序列进行生物信息学分析;利用实时定量PCR(qPCR)技术分析了PmBcl-2-like在马氏珠母贝不同组织、不同发育时期以及不同免疫刺激后的表达水平。结果显示,PmBcl-2-like cDNA全长为2 180 bp,开放阅读框长度为1 650 bp,共编码549个氨基酸,分子量为21.62 ku;结构域预测分析表明PmBcl-2-like含有Bcl-2家族典型的BH1-4结构域;多序列比对以及进化树构建结果表明,PmBcl-2-like与其他物种的相似度较高,保守性较强;荧光定量结果表明PmBcl-2-like在马氏珠母贝8个组织中均有表达,在鳃中表达量最高,其次为性腺,表达量最低为中央膜区;在胚胎期表达量较高,受精卵时期表达量最高。机体受到脂多糖(LPS)刺激后,相对表达量在24 h达到最高,72 h降到最低,最高约为最低的2.5倍;肽聚糖(PGN)刺激后,相对表达量在6 h达到最高,48 h降到最低,最高约为最低的7.28倍;聚肌胞苷酸(Poly:IC)刺激后,相对表达量在3 h达到最高,12 h降到最低,最高约为最低的6.49倍。研究表明,PmBcl-2-like可能在马氏珠母贝发育和免疫防御反应中担任着重要的角色。     

Detection of Minchinia occulta in samples of pearl oysters Pinctada maxima infected by Haplosporidium hinei

Objective   To determine if juvenile pearl oysters ( Pinctada maxima ) infected with Haplosporidium hinei are also infected with another haplosporidian parasite, Minchinia occulta .
Design   Archived samples of pearl oysters infected with H. hinei were examined using polymerase chain reaction (PCR) assays and in situ hybridisation (ISH) to analyse and identify haplosporidians. A 144-bp and 220-bp region of Minchinia DNA were targeted by PCR and amplified DNA from formalin-fixed H. hinei -infected pearl oyster samples was sequenced. A 25-bp oligonucleotide probe targeting a variable section of the parasite's small subunit rRNA gene was used in ISH.
Results   The results of DNA-based diagnostic assays supported each other. The sequences obtained by PCR were found to be almost identical to M. occulta from rock oysters and the ISH assay demonstrated infection with M. occulta in affected pearl oysters. ISH indicated a prevalence of infection of 26.7% in one of the previous outbreaks.
Conclusion   Pearl oyster spat are susceptible to infection by a Minchinia parasite, most likely M. occulta , which was recently identified in rock oysters within the pearl-producing zones of Western Australia and is associated with mortalities of up to 80% in this species. The occurrence of haplosporidian co-infections in pearl oysters suggests the immunocompetence of juvenile oysters may be an important factor in preventing infection and therefore preventing mortalities such as those occurring in the recent outbreaks of pearl oyster oedema disease.     

栅栏技术优化即食调味珍珠贝肉工艺的研究

文章运用栅栏效应的理论，确定了高水分型即食调味珍珠贝肉食品的制作工艺，分析了各种常见栅栏因子对制品感官品质及微生物的影响，优化前处理、烘干工艺、杀菌工艺、pH、水分活度（activity water，Aw）和低温处理等多种栅栏因子，形成有效防止制品腐败变质的栅栏模式。试验结果表明，原料前处理需用3％的食盐进行了清洗；采用0．15％柠檬酸溶液调节pH为5．6～5．7；贝肉调味后需经烫煮；产品水分含量45％～50％，Aw为0．88～0．90；产品真空包装，低温处理24h，再采用巴氏杀菌（80～85℃、30min），能较好地保持产品的品质和风味，延长保存期，保持珍珠贝肉特有的鲜味和营养价值，促进珍珠贝肉的开发利用。     

马氏珍珠贝肉营养液的研制及营养评价

采用酶技术,利用珍珠养殖场开珠后的废弃物－－马氏珍珠贝肉,研制珍珠贝肉营养液。结果显示,采用枯草杆菌中性蛋白酶和风味酶Ｆｌａｖｏｕｒｙｚｍｅ联合水解马氏珍珠贝肉的效果最好,通过正交试验法确定了酶法水解的最佳工艺条件是：温度５０℃,时间５ｈ,ｐＨ６．５,枯草杆菌中性蛋白酶和Ｆｌａｖｏｕｒｙｚｍｅ的添加量分别为０．６％和０．２５％。对营养液的营养价值进行分析评价,营养液中蛋白质达到７３．６ｍｇ／ｍＬ,     

人工神经网络优化酶法制备马氏珠母贝 低聚肽的工艺研究

以马氏珠母贝全脏器为原料,采用胃蛋白酶、胰蛋白酶以及胰凝乳蛋白酶模拟消化道酶解的方式制备低聚肽,先通过胃蛋白酶酶解获得初产物,在此基础上利用人工神经网络对胰蛋白酶及胰凝乳蛋白酶的双酶酶解过程进行模拟预测,对其酶解工艺进行优化.试验结果表明,通过人工神经网络预测获得小肠消化蛋白酶双酶酶解的优化工艺条件为:复合酶添加量4 500 U/g(m 胰蛋白酶∶m胰凝乳蛋自酶=6∶1)、料水比1∶2(W/V)、酶解时间3.5 h,在此条件下制备的低聚肽(200～2 000 u)得率达55％以上.通过优化的工艺条件制备的低聚肽可获得较高的得率,能够为马氏珠母贝全脏器低聚肽的开发利用奠定基础.     

术前饥饿处理对马氏珠母贝血淋巴免疫机能的影响

在室内养殖条件下研究了术前限食处理对马氏珠母贝血淋巴细胞密度、超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、溶菌酶(LSZ)、酸性磷酸酶(ACP)和碱性磷酸酶(AKP)活性的影响。实验设置每天投喂一次和两天投喂一次两个处理组,每天投喂两次为对照组,分别在实验开始和第1、2、3和4周取样检测血淋巴免疫功能。结果显示:限食1周后,马氏珠母贝血细胞密度、SOD、CAT、LSZ、ACP和AKP等酶活性均随限食程度的增加呈下降趋势,2个限食处理组的免疫参数较对照组明显下降,并表现出显著性差异(P0.05),两天投喂一次处理组的各项免疫机能均显著低于对照组,且这种趋势一直延续至第4周。据此推断,限食可以显著降低插核手术贝的生理机能和免疫活性,为马氏珠母贝育珠术前处理提供理论依据。     

海南珠母贝属Pinctada6个种的分类鉴定

通过测试海南省珠母贝属Pinctada 6个种的45个个体的16S和H3基因序列,并与GenBank数据库中相同物种进行比对,构建系统发育树对其进行分类,同时根据简单、明确的贝壳形态学特征予以最终确认。此分类法不仅可以较准确、快速地进行珠母贝属的分类鉴定,而且分类结果不易受被测试贝类生活环境的影响,不易受分类鉴定人员水平的限制,有利于发现杂交种和隐存种。