Nitrogen and phosphorus budget of a Haliotis discus hannai and Apostichopus japonicus polyculture system

The system nitrogen (N), phosphorous (P) budget, N/P utilization rate and the physiological response mechanism of the abalone Haliotis discus hannai (body weight: 12.87 ± 0.82 g) and the sea cucumber Apostichopus japonicus (body weight: 10.85 ± 1.16 g) to different co‐culture environment conditions were examined. Animals were kept in a multilayer, cubic recirculating aquaculture system at different polyculture densities (abalones at 400 ind/m²in monoculture [Group C] and abalones at 400 ind/m²with sea cucumbers at 10 ind/m² [AS1] or 20 ind/m² [AS2]). Each treatment was replicated four times, and the experimental cycle was 90 days. No significant difference in survival rate of abalones was detected when the stocking density of sea cucumbers increased from 10 to 20 ind/m², but the concentrations of total ammonia nitrogen and nitrite nitrogen in the water were significantly higher in AS2 than in AS1. Survival rate, specific growth rate (SGR) of body weight of sea cucumbers and SGR of body weight of abalones were significantly lower in AS2 than in AS1. No significant difference in protease (PES), lipase, amylase and cellulase activities of abalones was identified between Group C and AS1, but the PES and amylase activities of abalones and sea cucumbers in AS1 were significantly higher than those in AS2. In AS1, the N/P output from harvesting of abalones and sea cucumbers and the N/P utilization rates were significantly higher than those in AS2. Although the N/P output from faeces was significantly lower in AS2 than in Group C and AS1, the N/P output from the water layer was significantly higher than that in AS1. The expression levels of Cu/Zn superoxide dismutase (Cu/Zn‐SOD) and heat shock protein 70 (HSP70) of both abalones and sea cucumbers in AS2 were significantly higher than those in AS1. No significant difference in expression of catalase (CAT) and HSP90of abalones was identified among these groups, but the expression levels of CAT and HSP90of sea cucumbers in AS2 were significantly higher than those in AS1. These results indicate that stocking sea cucumbers at 10 ind/m² in the polyculture system will relieve the organic load on the system and improve the N/P utilization rate. It will also increase aquaculture production and improve the ecological and economic benefits of the system.     

基于DEB理论的皱纹盘鲍个体生长模型参数的测定

为获取皱纹盘鲍(Haliotis discus hannai)个体生长模型所需的6个关键参数,设计了饥饿耗能、温度对耗氧的影响等相关生理实验,计算得到各项参数值。单位体积维持耗能率的值 、形成单位体积结构物质所需的能量值 、单位体积最大储存能量 和储备能量值 4个参数,通过连续测定皱纹盘鲍饥饿过程中呼吸耗氧率和软组织干重不断下降直至保持稳定时的能量值计算;温度函数中Arrhenius温度 的数值根据皱纹盘鲍在不同温度梯度下的单位干重耗氧率测定、计算;形状系数δm值通过统计测量的壳长、软组织湿重等生物学参数拟合回归得到。结果显示,皱纹盘鲍在饥饿后,呼吸耗氧率和软组织干重分别降低了26.3%和70.0%,呼吸耗氧率由2.69 mg/(ind.?h)逐渐降低并稳定在0.8 mg/(ind.?h),软组织干重由(5.21±0.89) g降低至(3.84±0.22) g;根据公式计算得 和 的值分别为20.18 J/(cm3?d)和8120 J/cm;皱纹盘鲍饥饿前后有机物含量分别为80%和58%,经过换算, 和 的值分别为2726 J/cm3和32583 J/g。不同规格的皱纹盘鲍在水温为5℃~20℃范围内,温度与单位干重耗氧率呈正比;当水温超过20℃之后,温度与单位干重耗氧率呈反比。在转折点20℃之前,单位干重耗氧率的ln值与温度(热力学温度,K)的倒数呈线性关系,线性回归方程斜率的绝对值为Arrhenius温度 值( =7196 K)。生物学统计分析鲍壳长(L)与体积(V)呈三次函数关系：V=0.0639 L3.1621(R²=0.9852),根据公式对软组织湿重的立方根与壳长进行线性回归,所得的斜率即为形状系数δm值(δm=0.43)。本研究对建立以DEB理论为指导的皱纹盘鲍个体生长模型提供了数据支撑。     

Microsatellite pedigree analysis reveals high variance in reproductive success and reduced genetic diversity in hatchery-spawned northern abalone

The northern (or pinto) abalone, Haliotis kamtschatkana, is a broadcast-spawning marine gastropod that was recently listed as endangered in Canada. To aid in species recovery, a captive-breeding and supplementation program is underway in Barkley Sound, British Columbia. We genotyped first generation progeny for five microsatellite loci and used a pedigree reconstruction program (Pedigree 2.2) to identify their genealogical relationships in the absence of information on parental genotypes. We analyzed progeny from three separate group-spawning events and inferred considerable variation in the number of offspring produced by each parent; in the most severe case a single male sired all the progeny produced during one spawning event. After only one generation of captive-breeding we found a 55–60% reduction in allelic richness and a 17–18% reduction in heterozygosity relative to the diverse wild source population. This study illustrates the difficulty of managing genetic diversity in hatchery populations of a broadcast-spawning species, even when gametes are collected separately from each individual broodstock.     

皱纹盘鲍与盘鲍杂交效果分析

从杂种优势的角度对皱纹盘鲍♀与盘鲍♂杂交子一代的经济性状进行分析。结果表明,4月龄杂交鲍子一代(F1)活体重比皱纹盘鲍增加36.87%,软体部干重增加33.21%,总干重增加29.20%,其它性状也显著优于皱纹盘鲍。杂交鲍F1的壳长、壳宽、活体重、软体部干重和总干重等主要性状的变异系数为12.94%~44.51%,其中软体部干重的变异系数最大,壳长的变异系数最小;且杂交鲍各性状的变异系数均大于皱纹盘鲍。杂交鲍子一代的变异有助于定向选择,提高杂交育种的效果。     

不同温度对九孔鲍能量收支影响的研究

在摸拟自然条件下,设定5个温度梯度（15、19、23、27、3lac）进行了室内水族箱养殖,结果表明,九孔鲍（Haliotis diversicolor superrtesxa）的生长能和代谢能随温度的增加而增加,高于此温度时则相反,呈负相关关系。在温度为27℃时,九孔鲍的生长能占摄食能的比例达到最大值。九孔鲍在各温度下的能量收支方程为：15℃：100（C）=18．05（F）＋8．00（U）＋7．91（M）＋39．60（R）＋7．14（Psh）＋9．36（Pg）＋9．94 19℃：100（C）=19．89（F）＋12．24（U）＋7．97（M）＋33．74（R）＋6．62（Psh）＋9．54（Pg）＋10．01 23℃：100（C）=17．43（F）＋12．93（U）＋7．04（M）＋33．79（R）＋6．20（Psh）＋10．25（Pg）＋12．37 27℃：100（C）=15．56（F）＋11．03（U）＋7．05（M）＋28．53（R）＋13．67（Psh）＋16．12（Pg）＋8．03 31℃：100（C）=18．82（F）＋13．87（U）＋12．52（M）＋38．44（R）＋12．35（Psh）＋13．62（Pg）-9．62从能量收支方程中看出,只有小部分能量用于生长壳,一部分能量用于生长软体肉能,绝大部分能量用于代谢,还有一小部分能量用于排泄、排粪和分泌粘液。     

皱纹盘鲍4个群体与九孔鲍16S rRN A基因遗传差异分析

采用PCR方法扩增了皱纹盘鲍（4个群体）和九孔鲍（1个群体）共5个群体（53个样本）16S rRNA基因片段（16S）,测序获得了1000 bp核苷酸序列,其中669 bp核苷酸序列用于遗传差异分析,旨在评估我国皱纹盘鲍不同群体间及其与九孔鲍间的遗传差异。从52个序列中共检测到17种单倍型,其中皱纹盘鲍4个群体有13种单倍型,九孔鲍1个群体（9个样本）有4种单倍型。单倍型核苷酸序列比对显示,变异位点占比对位点的20．5％,基于单倍型的皱纹盘鲍4个群体间的遗传距离为0．002～0．011,平均为0．005;九孔鲍群体内的平均遗传距离为0．004,两种鲍间的平均遗传距离为0．218;两种鲍间的遗传分化系数 F‐统计量 FST平均为0．982,皱纹盘鲍4群体间的 FST值为0．0051～0．0065。本研究显示2种鲍群体内16S变异很小。系统发育分析鲍属11种鲍的聚类关系,显示皱纹盘鲍与盘鲍、日本鲍交叉聚为一支,九孔鲍与杂色鲍也交叉聚为一支,4个种类未聚成单系支。     

杂色鲍同种移植炎症因子1的克隆及其在应激下的表达

同种移植炎症因子AIF-1 (allograft inflammatory factor 1,AIF-1)是一种由干扰素γ诱导的含有EF-hand结构域的钙离子结合蛋白,其功能主要是参与移植排斥、免疫炎症反应、自身炎性和非炎性的损伤等.首次克隆了杂色鲍AIF-1基因cDNA全序列,命名为HdAIF-1,其全长为942 bp,开放阅读框为456 bp,编码151个氨基酸.实时荧光定量PCR结果表明:HdAIF-1在杂色鲍各组织中均有表达,其中在血淋巴和鳃中表达量最高.高温应激下,HdAIF-1在鳃组织中各时相表达均显著上调,并在温度升至31℃时达到最高.而血淋巴和肝胰腺中HdAIF-1在高温应激前4个时相表达无显著差异,到了96 h均显著上调.缺氧应激下,HdAIF-1在血淋巴中表达变化没有显著差异,而鳃中24 h显著下调,192 h显著上调.副溶血弧菌感染实验表明HdAIF-1基因在感染后3、24和48 h均检测到HdAIF-1的表达量显著上调.高温和缺氧应激以及弧菌感染均显示HdAIF-1基因表达量发生显著变化,说明HdAIF-1可能作为免疫因子在杂色鲍应激等状况下发挥重要作用.     

杂色鲍群体间杂交的杂种优势分析

杂色鲍（Haliotis diversicolor）是中国南方主要的鲍养殖种类,近年来“东优1号”杂色鲍因较高的成活率优势已在产业上得到越来越广泛的推广养殖.该研究采用3×3完全双列杂交的方式,对杂色鲍“东优1号”群体、杂色鲍古雷群体和杂色鲍越南群体3个群体自繁与杂交后代的养殖性能进行跟踪,并对其杂种优势进行评估.结果表明,3个群体自繁后代的生长速度和成活率上存在显著差异（P＜0.05）,同时3个组合的生长速度与成活率的双亲杂种优势率值均不高,且2个单亲杂种优势率值之间有显著差异（P＜0.05）.     

九孔鲍选择群体F1的选择反应与现实遗传力估计

以中国九孔鲍（Haliotis diversicolor aquatili）养殖群体与日本野生群体杂交子二代为选育基础群体,采用群体选育方法进行九孔鲍的选择育种研究,以估计该养殖群体的选择反应与现实遗传力。截断选择壳长最大的10％个体作为选择组亲本（SS）,从基础群体中随机抽取相同数量的个体作为对照组亲本（sc）,选择强度为1．755,结果表明,选择组与对照组在卵径、受精率方面无显著的差别（P〉0．05）,但选择组在幼体附着率、幼体变态率、幼体成活率及稚贝早期生长方面与对照组相比表现出不同程度的生长优势（P〈0．05）,选择组50日龄的稚鲍成活率达到35．07％,显著大于对照组23．62％（P〈0．05）,40—270日龄的选择组壳长选择反应和现实遗传力变化范围分别为13．64％～81．88％和0．21～0．53,平均为31．63％和0．36,选择组平均壳长显著大于对照组（P〈0．01）。研究初步显示,养殖群体的选择反应极显著,群体选育是养殖九孔鲍遗传改良的有效途径。     

中国海耳鲍成熟卵的超微结构

利用光学和电子显微镜对耳鲍排出体外成熟卵的形态及超微结构进行了观察。耳鲍成熟卵具有卵胶质膜、卵黄膜和细胞膜;卵质中含有线粒体、核糖体、内质网及高尔基体等细胞器;还含有高电子密度的分泌颗粒和吞饮小泡。皮层中含有皮质小泡,但未发现中心粒。耳鲍成熟卵没有明显的卵极标志,卵黄颗粒分布较均匀。