镉和苯并芘胁迫对文蛤血细胞功能的影响

应用流式细胞仪技术,通过检测血细胞的吞噬活力、活性氧产物和死亡率,研究了Cd2+和苯并芘(Bap)胁迫7d对文蛤血细胞功能的影响。结果表明,10μg/L和100μg/LCd2+胁迫会显著降低血细胞的吞噬活力,增加血细胞的死亡率,100μg/LCd2+会显著增加血细胞活性氧产物。5μg/L和50μg/LBap胁迫会显著降低文蛤血细胞的吞噬活力,增加血细胞的死亡率,50μg/LBap会显著增加血细胞活性氧产物。可见,Cd2+和苯并芘胁迫会显著降低文蛤血细胞的功能,导致文蛤免疫力下降,增大文蛤被病害感染的机率。     

文蛤一种球形病毒的分离纯化及细胞病理变化观察

为研究一种球形病毒引起的文蛤(Meretrix meretrix)细胞病理变化及该病毒的分类地位,作者运用透射电镜对异常死亡文蛤的细胞病理变化及感染的病毒粒子进行了超微结构观察,并通过差速离心、酒石酸钾超速离心和氯化铯密度梯度离心结合等方法对首次对该病毒粒子进行了分离纯化。结果表明:患病个体的鳃、消化盲囊、消化道和外套膜均大量存在一种病毒颗粒,该病毒呈球形,无囊膜,直径45~55 nm,核心与核衣壳间无明显界限,未发现病毒包涵体,该病毒存在于细胞间质及被破坏的细胞质中。该病毒粒子的感染引起的细胞病理变化主要表现为:细胞核染色质集聚,溶酶体增多,线粒体肿涨,粗面内质网膨胀,肌原纤维排列混乱,细胞结构变得无规则直至解体。病毒粗提液于4℃150 000 g氯化铯密度梯度离心20 h可该病毒粒子提纯,经纯化后负染显示,该病毒粒子呈球形,二十面体对称,无囊膜,直径在60~75 nm,表面结构粗糙,壳粒结构较明显,该病毒粒子在氯化铯中的浮密度为1.554~1.582 g/m L。本文结果为文蛤死亡病原的研究提供了基础资料。     

氨氮对文蛤存活及能量收支的影响

在实验室条件下,研究了氨氮(NH3-N)对文蛤(Meretrix meretrix)的急性毒性作用以及对其能量收支的影响。实验结果表明:在水温(24.5±0.5)℃,盐度21.5,pH8.00的条件下,文蛤(壳长3.85～4.35cm)死亡率随着氨氮质量浓度、胁迫时间的增加而增加(P0.05)。96 h文蛤死亡率与添加氨氮(NH3-Nt)质量浓度对数的回归方程为Y=2.3787x+0.3246(R2=0.978 9),文蛤对总氨氮(NH3-Nt)或分子氨(NH3-Nm)的安全浓度(SC96h)为9.237 mg/L或0.418 mg/L;随着环境中添加氨氮(NH3-Nt)质量浓度的不断上升(0,0.5,1,2,4,9,18,36 mg/L),文蛤摄食能不断下降,呼吸能和排泄能则呈现出先升高后降低的趋势;当总氨氮(NH3-Nt)在0～2 mg/L(或NH3-Nm≤0.090 mg/L)时,文蛤的生长余力没有显著变化(P﹥0.05);当总氨氮(NH3-Nt)≥4 mg/L(或NH3-Nm≥0.181mg/L)时,文蛤的生长余力显著降低(P0.05)。因此,文蛤养殖环境中的NH3-Nt或NH3-Nm应控制在2mg/L或0.090 mg/L以下水平。     

4个文蛤群体鲜味物质比较分析

以江苏文蛤(Meretrix meretrix)为研究对象,采集红壳色文蛤原种、黄壳色文蛤原种、红壳色文蛤选育F1、红壳色文蛤选育F2 4个群体,检测其主要非挥发性鲜味物质成分单磷酸腺苷(AMP)、单磷酸鸟苷(GMP)、次黄嘌呤核苷酸(IMP)、琥珀酸、游离氨基酸、无机离子(Na~+、K~+、Cl~–、PO_4~(3–))的含量,并通过味道强度值(TAV)评价其呈味作用。鉴于核苷酸与氨基酸在呈味方面的协同效应,采用味精当量(EUC)评价不同文蛤群体的鲜味品质。结果表明,文蛤软体组织中AMP、琥珀酸、谷氨酸、精氨酸、丙氨酸、Na+、K~+、Cl~–的TAV值大于1,是文蛤鲜味的主要贡献者;红壳色文蛤原种的鲜味强度最大(4.92 g/100g),其子代红壳色文蛤F1(4.08 g/100g)、红壳色文蛤F2(4.09 g/100g)稍有降低,但仍显著高于黄壳色文蛤原种(3.34 g/100g)(P0.05),表明江苏红壳色文蛤在鲜味品质方面具有相对稳定的较高品质。     

利用父母本微卫星标记估计大菱鲆(Scophthalmus maximus L.)幼鱼生长性状的遗传参数

The estimation of genetic parameters has played an important role in animal selective breeding for growth traits.Recently studies show that molecular markers can be incorporated into genetic evaluations. In order to improve the performance of an incomplete pedigree(i.e, only parents are known) in the genetic evaluations, 12 microsatellite markers have been applied in the estimation of the genetic parameters for body weight in a farmed population(n=1 890) of juvenile turbot(Scophthalmus maximus L.). A new relatedness called parental molecular relatedness(PMR) is estimated based on results of genotyping of 48 parents(31 males, 17 females) with microsatellites markers. The feasibility of PMR in estimation of genetic parameters is verified by comparison with pedigree related(PR) which is obtained from a complete pedigree. The results demonstrate that a high correlation(0.872) between them is found. Heritabilities are estimated using the PMR(0.52±0.13) and PR(0.55±0.22) with the same animal model. A cross-validation shows that the predictive abilities of models using the PMR and the PR are identical(0.81). From that, a conclusion can be made that PMR and PR predicted genetic values equally well in a population of juvenile turbot. Therefore PMR can be applied as an alternative of the PR when only parents are known. However, for a better performance, more markers and more families should be included in a further study.     

皱肋文蛤胚胎、幼虫及稚贝的发育

在水温28.5～30℃、盐度23.6～25.4 条件下, 观察了皱肋文蛤(Meretrix lyrata)胚胎、幼虫和稚贝的发育过程。结果表明, 皱肋文蛤的胚胎经历桑椹期、囊胚期、原肠胚期等阶段, 形成担轮幼虫。根据行为方式和形态特征, 其幼虫阶段可分为D 形幼虫、壳顶初期幼虫、壳顶中期幼虫、壳顶后期幼虫、变态期幼虫、附着期幼虫。稚贝发育可分为无管期稚贝、单管期稚贝和双管期稚贝。皱肋文蛤幼虫在不同阶段的生活习性逐渐改变, 从担轮幼虫至壳顶后期幼虫营浮游生活, 变态期和附着期幼虫分别营匍匐生活和附着生活。壳顶初期幼虫出现足原基, 壳顶中期和壳顶后期幼虫分别形成平衡囊和鳃丝。稚贝阶段开始从附着生活向埋栖生活过渡。稚贝出现明显的次生壳, 无管期稚贝和单管期稚贝的次生壳为红褐色, 而双管期稚贝的次生壳为白色。皱肋文蛤从D 形幼虫发育到附着期幼虫, 后者的壳长和壳高分别增至前者的1.77 倍和2.10 倍; 稚贝阶段从无管期发育到双管期, 后者的壳长和壳高分别增至前者的3.13 倍和3.28 倍, 显示皱肋文蛤幼虫阶段缓慢生长, 稚贝阶段快速生长的特征。     

红鳍东方鲀(Takifugu rubripes)耐低温性状和生长性状遗传参数评估

采用人工控温的方式, 对构建的31个F₁红鳍东方鲀(Takifugu rubripes)全同胞家系开展低温胁迫实验, 获得耐低温性状低温累计存活时间(CDH), 基于混合线性模型分别开展耐低温性状和生长性状遗传参数评估,对每一性状是否需要考虑共同环境效应所构建的两种模型进行似然比检验。结果显示, 经似然比检验, 最终选用模型A和模型BF进行耐低温和生长性状遗传评估; 耐低温性状CDH遗传力为(0.27±0.08),属于中等遗传力; 体重BW遗传力为(0.36±0.13), 属于中等遗传力, 体长BL遗传力为(0.14±0.06), 属于低等遗传力, 经检验, 遗传力估计值均达到极显著水平(P<0.01)。CDH和体重、体长的遗传相关分别为(-0.40±0.22)和(-0.44±0.24), 表型相关分别为(-0.09±0.06)和(-0.16± 0.05), 均为负相关; 体重和体长之间的遗传相关为(0.92±0.05), 表型相关为(0.80±0.02), 呈正相关且结果极显著(P<0.01)。研究结果表明, 红鳍东方鲀的耐低温性状和生长性状都具有较好的改良潜力, 考虑到两性状间存在负遗传相关, 在开展耐低温选育时, 对首先不同性状进行品系选育, 然后利用品系间杂交培育出耐高温、生长快的新品种。该项研究首次完成了红鳍东方鲀耐低温性状的遗传参数评估, 为制订红鳍东方鲀耐低温选育育种规划提供了理论依据。     

海水酸化和甲基汞胁迫条件下文蛤免疫应答和生物矿化响应

海洋酸化背景下重金属对海洋生物的毒性效应是一个重要的生态毒理学研究问题。海洋酸化不仅直接影响海洋贝类的生理过程,也通过改变重金属的存在形式和生物可利用性进而影响其生物毒性。为研究海洋酸化背景下甲基汞(MeHg)对海洋贝类免疫和生物矿化的毒理效应,本研究将采集于野外的文蛤(Meretrix petechialis)置于不同pH水平(二氧化碳分压;pH 8.10/背景水平、7.70/中度酸化和7.30/高度酸化)和甲基汞质量浓度(对照、溶剂对照、0.1,1和5 μg/L)的海水中共同暴露21 d,研究文蛤内脏团和鳃组织内免疫应答和生物矿化相关的生物标志物对海水酸化和MeHg共同胁迫的响应。结果表明,海水酸化和MeHg均显著影响其免疫应答策略,不同胁迫水平对各类生物标志物具有组织差异性。具体而言,MeHg暴露诱导内脏团中碱性磷酸酶(AKP)和溶菌酶(LZM)活性,表明MeHg对免疫解毒机制有刺激作用,在一定程度上提高了其免疫应答。海水酸化抑制了鳃和内脏团中AKP活性,抑制其免疫应答。在生物矿化相关酶中,在海水酸化和MeHg共同胁迫下钙-ATP酶(Ca²⁺-ATP)活性显著下降,干扰其离子平衡和生物矿化。海水酸化加剧了MeHg胁迫对文蛤免疫应答和生物矿化的毒理作用。相关系数分析和主成分分析表明这些生物标志物可以协同防御环境胁迫对免疫应答和生物矿化策略的毒理作用。这些生物酶对海水酸化和MeHg胁迫响应比较敏感,可以作为评价海洋酸化背景下MeHg对文蛤两种生物组织免疫功能毒性效应的潜在生物标志物。研究结果为理解海洋酸化和重金属胁迫对贝类生理功能的影响提供新见解,为评估海洋酸化背景下贝类种群变动和资源管理提供科学依据。     

施氏獭蛤早期生长性状遗传参数和育种值估计

以2代选育群体为亲本、采用巢式设计构建21个施氏獭蛤(Lutraria sieboldii)全同胞家系,利用混合线性模型对30日龄和80日龄施氏獭蛤生长性状进行遗传参数和育种值估计.结果显示:施氏獭蛤早期发育过程中壳长生长速度较壳高快,性状间遗传相关系数范围为0.96～0.98,表型相关系数范围为0.77～0.96,均...     

不同盐度对文蛤(Meretrix meretrix)呼吸代谢及体内酶活性的影响

为研究不同盐度对文蛤呼吸代谢的影响,本实验设置5个盐度(‰)梯度(11、18、25、32、39),检测不同盐度对文蛤(Meretrixmeretrix)耗氧和排氨的影响,以及文蛤的外套膜、鳃、肝胰腺三种组织中乳酸脱氢酶和Na+/K+-ATP酶活性的变化。结果表明:随着盐度的不断升高,文蛤耗氧率先升后降再升,在盐度18时达到最大值;排氨率先升后降,在盐度32时达到最大值。随着盐度不断升高和胁迫时间延长,文蛤的肝胰腺中乳酸脱氢酶活力总体呈先升高后下降再升高的趋势(P0.05),酶活力在盐度39时为最高;随着盐度不断升高和胁迫时间延长,文蛤的外套膜中Na+/K+-ATP酶活力总体呈先下降再升高后下降的趋势(P0.05),在盐度32时为最高;文蛤的外套膜和鳃中乳酸脱氢酶活力以及鳃和肝胰腺中Na+/K+-ATP酶活力受盐度影响不显著(P0.05),酶活力变化也多呈现"W"形的变化趋势。研究结果为文蛤的人工养殖提供参考。     

因子交配设计中文蛤亲本数目对遗传参数估计的影响

The precise and accurate knowledge of genetic parameters is a prerequisite for making efficient selection strategies in breeding programs.A number of estimators of heritability about important economic traits in many marine mollusks are available in the literature,however very few research have evaluated about the accuracy of genetic parameters estimated with different family structures.Thus,in the present study,the effect of parent sample size for estimating the precision of genetic parameters of four growth traits in clam M.meretrix by factorial designs were analyzed through restricted maximum likelihood(REML) and Bayesian.The results showed that the average estimated heritabilities of growth traits obtained from REML were 0.23–0.32 for 9 and 16 full-sib families and 0.19–0.22 for 25 full-sib families.When using Bayesian inference,the average estimated heritabilities were0.11–0.12 for 9 and 16 full-sib families and 0.13–0.16 for 25 full-sib families.Compared with REML,Bayesian got lower heritabilities,but still remained at a medium level.When the number of parents increased from 6 to 10,the estimated heritabilities were more closed to 0.20 in REML and 0.12 in Bayesian inference.Genetic correlations among traits were positive and high and had no significant difference between different sizes of designs.The accuracies of estimated breeding values from the 9 and 16 families were less precise than those from 25 families.Our results provide a basic genetic evaluation for growth traits and should be useful for the design and operation of a practical selective breeding program in the clam M.meretrix.     

硒化物在文蛤响应镉胁迫过程中作用的转录组学分析

研究两种硒化物在文蛤富集、排出镉过程中的作用,对文蛤进行转录组测序,并进行生物信息学分析,以探讨硒化物对Cd2+代谢相关机制。硒化卡拉胶组和亚硒酸钠组共产出587 855条高质量reads。基因本体分析功能注释到15 380条unigenes, KEGG通路注释到18 866条unigenes。差异基因主要富集到肌球蛋白复合物(myosin complex)、离子通道抑制(ion channel suppression)等基因本体分析功能中。差异基因KEGG通路富集显示,在嘌呤代谢(purinemetabolism)、ECM受体相互作用(extracellularmatrix receptor interaction)、硒化合物代谢(metabolism of selenium compounds)等通路显著富集。从结果分析Cd2+可以使文蛤体内蛋白转录修饰出现异常;有机硒可以通过调控金属离子通道活性来排出细胞内的Cd2+;无机硒主要作用于文蛤细胞表面,增强其表面活性抵御Cd2+进入细胞;差异基因KEGG注释结果表明有机硒与无机硒在文蛤重金属代谢中作用机制以及途径不相同。     

南黄海沿岸滩涂文蛤中痕量硒的周年变化探讨

采用4︰1混合酸湿法消解,氢化物发生原子荧光光谱法(HG-AFS)测定了南黄海沿岸滩涂文蛤生物体中微量元素硒的周年变化,并用质控样和加标回收保证结果的可靠性。结果显示:硒的周年含量为0.2~0.5 mg/kg,平均值为0.330 mg/kg,文蛤体内硒的含量呈一定规律性变化,春、秋季高,夏、冬季低。说明文蛤中硒含量因不同季节而具有明显差异,其含量变化与海水温度、饵料生物及文蛤生长繁殖等因素有关。     

文蛤亮氨酸氨肽酶LAP3的基因克隆及其在幼体不同发育时期、成体不同组织的表达分析

为探索贝类LAP3基因的结构和功能特征以及在生长发育过程中的作用,利用cDNA末端快速扩增（RACE）法获得文蛤LAP3（Mm-LAP3）基因的cDNA全长序列,并对其生物信息学、组织及发育时期表达特征进行了分析。结果显示,Mm-LAP3基因cDNA全长2 037 bp,ORF区1 254 bp,编码417个氨基酸;Mm-LAP3蛋白由Peptidase_M17超家族序列的N-端结构域和Peptidase_M17催化结构域组成。荧光实时定量PCR结果表明,Mm-LAP3基因在文蛤成体6个组织和幼体10个发育时期中均有表达,其中在斧足中的表达量显著高于其他组织（P<0.05）,这与斧足蛋白含量丰富、代谢旺盛相关;在幼体10个发育时期中的表达量呈现逐渐升高的趋势,在D形幼虫时期达到最高,随后又有所降低,推测Mm-LAP3基因在早期发育时期参与了某些组织器官的形成。     

三个文蛤家系双列杂交设计在两种环境下的配合力及杂种优势分析

A 3×3 complete diallel cross comprising three families of the clam Meretrix meretrix(P1, P2 and P3) was used to determine the combining ability of parental families and heterosis of F1 under indoor and openair environments for growth traits. Analysis of variance for shell length and whole body weight indicated highly significant cross effects, environment effects and the interaction of cross by environment. General combining ability(GCA) and specific combing ability exhibited great variation among crosses and between two environments. Pooled over environments, P2 was the top combiner among the three parental families for both traits studied. The cross of P1 and P3 had the highest SCA. Additionally, significant reciprocal effects were observed. For individual environment, about half of the crossbred combinations showed favorable Mid-parent heterosis(MPH)(1%) for the shell length and whole body weight. Our data has shown that non-additive genetic and reciprocal effects constituted the major sources of genetic variation for both shell length and whole body weight, which indicates that crossbreeding among selective families could further explore the heterotic effects.     

Genetic parameter estimation for juvenile growth and upper thermal tolerance in turbot (Scophthalmus maximus Linnaeus)

Twenty-six half-sib groups(53 full-sib families) of turbot, Scophthalmus maximus Linnaeus, were obtained by artificial insemination. We measured growth in the offspring(40–50 individuals/family) and subjected them to a thermal tolerance challenge over a period of 34 d. There was no significant difference in daily mortality(range: 0.580%–1.391%) between Days 1–13 during the thermal tolerance challenge. However, daily cumulative mortality increased rapidly between Days 14 and 29, especially on Days 15 and 16(20.232% and 34.377%, respectively). Mortality was highest on Day 16(14.145%). We estimated the genetic parameters using the average information restricted maximum likelihood method. We used a likelihood ratio test to evaluate the significance of effects in models with and without identity as an effect, and compared the final log-likelihoods(maximum log L). Lastly, we estimated phenotypic and genetic correlation between the upper thermal tolerance limit(UTT) and body weight(BW). In this study, the positive phenotypic correlation was low between UTT and BW(0.093±0.029). The genetic correlation between UTT and BW was negative(-0.044±0.239). The heritability for upper thermal tolerance was low(0.087±0.032), which is of approximately moderate heritability. The heritability for body weight was high(0.303±0.074). Our results suggest there is significant potential for improvement in the culture of turbot by selective breeding.