红螯螯虾养殖群体遗传多样性的SSR分析

本研究利用10个微卫星标记对广东和台湾红螯螯虾(Cherax quadricarinatus)养殖群体进行遗传多样性分析,并对2个红螯螯虾群体的平均等位基因数(Na)、平均有效等位基因数(Ne)、观测杂合度(Ho)、期望杂合度(He)等进行计算.结果显示:在2个红螯螯虾群体中共检测到83个等位基因,广东群体的平均有效等...     

不同光周期下红螯螯虾幼虾摄食节律的研究

在不同光周期下对红螯螯虾(Cherax quadricarinatus)幼虾的摄食节律进行了研究。结果显示:红螯螯虾具有明显的日摄食节律,24 L(持续光照)、12 L∶12 D(12 h光照:12 h黑暗)和24 D(持续黑暗)光周期下,其摄食率、摄食量和摄食发生率均在19∶00-21∶00和5∶00-7∶00出现峰值,分别为:13.01%和15.58%、20.82%和14.75%、12.77%和16.89%,属晨昏型摄食。光周期和不同时段对红鳌幼虾摄食无显著的交互作用(P>0.05)。     

不同地区红螯螯虾(Cherax quadricarinatus)养殖群体遗传多样性分析

为了解国内广东省湛江市、广东省佛山市、广东省惠州市、广西自治区南宁市、浙江省湖州市5个地区红螯螯虾养殖群体的遗传结构,我们鉴定了红螯螯虾8个微卫星位点。利用8个微卫星标记对5个地区的红螯螯虾养殖群体进行了遗传分析,8对引物在150个样本个体中共检测出237个等位基因。所有8个微卫星的等位基因数范围为2～10,平均为6.05。H_o和H_e平均值分别为0.490和0.614。每组的等位基因数在6.000(广西南宁GXNN)到6.625(广东佛山GDFS)之间。所有群体中平均H_e在0.593(广东惠州GDHZ)到0.646(广东湛江GDZJ)之间。由此可以看出种群的遗传多样性并未显著降低。F_is、F_st值、分子方差分析(AMOVA)结果显示,这些红螯螯虾群体大部分的遗传变异来源于种群内部,并且各个群体都保持着较高水平的遗传距离,其中GDZJ和ZJHZ群体之间遗传距离最大(0.107 7)。推测导致这些差异的产生可能是随机遗传漂变和人工选择。     

急性氨氮胁迫对红螯螯虾血细胞凋亡的影响

氨氮是水产养殖过程中的常见毒性污染物之一,为探讨氨氮对红螯螯虾(Cherax quadricarinatus)血细胞的细胞毒性机制,研究了氨氮胁迫作用下红螯螯虾血细胞凋亡状态的变化。以17.6mg/L氨氮对红螯螯虾幼虾进行急性胁迫,于胁迫后的0、48、72和96h取血淋巴,应用流式细胞术测定血细胞总数(THC)、胞内游离Ca~(2+)含量、线粒体膜电位(MMP)和血细胞凋亡率。结果显示,红螯螯虾的THC在胁迫的72h开始有显著的下降,在96h时达到最低值;血细胞胞内游离Ca~(2+)含量在48h开始有显著的升高,在96h达到最高值;血细胞MMP在72h开始有显著的下降,在96h达到最低值;血细胞凋亡率在72h开始有显著的升高,在96h时达到最高值,为9.39%。这些结果表明,胞内游离Ca~(2+)含量是对氨氮胁迫最为敏感的指标;氨氮急性胁迫诱导红螯螯虾血细胞发生了Ca~(2+)参与调节、线粒体通路相关的细胞凋亡,从而导致了THC的下降。     

饲料蛋白质水平对红螯螯虾生长、消化酶和抗氧化系统的影响

为了探讨饲料不同蛋白质水平对红螯螯虾(Cherax quadricarinatus)生长及生理生化的影响,按照等能量不等蛋白的原则,配置蛋白质水平分别为20%、24%、28%、32%、36%、40%的6组饲料投喂初始体重为(0.35±0.07)g的红螯螯虾幼虾60 d,然后测定各相关指标。存活率随着饲料蛋白质水平的增加逐渐升高(P<0.05)。当饲料蛋白质水平为28%时,特定生长率、肥满度最高,饲料系数最低。饲料不同蛋白质水平对红螯螯虾胃中的蛋白酶活性影响显著(P<0.05),胃蛋白酶活力先升高后下降,32%组活性最大。肝胰腺中的淀粉酶、谷草转氨酶(GOT)、过氧化物歧化酶(SOD)活性变化不显著(P>0.05)。谷丙转氨酶(GPT)活性随着蛋白质水平的增加逐渐升高(P<0.05)。过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GPX)活性先升高后降低,腮Na+-K+ATP酶则相反(P<0.05)。36%组和40%组的丙二醛(MDA)含量最高,其次为20%组,均显著高于其他各组(P<0.05)。结果表明红螯螯虾对饲料蛋白质含量的要求较低,28%即可满足其要求。豆粕含量过高可能会影响其生长。     

稻田养殖红螯螯虾个体大小与肠道菌群相关性研究

为了解稻田养殖红螯螯虾(Cherax quadricarinatus)个体大小与肠道菌群的相关性,实验采用16S rDNA高通量测序和生物信息学技术对稻田养殖红螯螯虾大个体(RB)组和小个体(RS)组肠道菌群的组成、多样性、指示物种、功能等进行比较分析.结果显示:RB组共产生615个OTUs(Operational t...     

青虾冷休克蛋白Y-box编码基因的cDNA全长克隆与表达分析

为研究冷休克蛋白Y-box基因在青虾应答环境胁迫过程中所起的调控作用,实验应用RACE PCR技术首次克隆了青虾的冷休克蛋白Y-box基因全长c DNA序列,并利用在线软件对其序列特征进行生物信息学分析;采用实时荧光定量PCR技术对其在青虾不同组织及环境胁迫过程中的表达变化特征进行分析。青虾冷休克蛋白Y-box基因c DNA全长1501 bp,包括84 bp的5′末端非翻译区(UTR),876 bp的开放阅读框(ORF),541 bp的3′UTR,开放阅读框编码291个氨基酸。氨基酸相似度比对显示,青虾冷休克蛋白Y-box基因富含高度保守的冷休克结构域。系统进化树分析显示,青虾冷休克蛋白Y-box基因与水蚤等节肢动物冷休克Y-box聚类一支,具有最近的亲缘关系。荧光定量PCR检测显示,冷休克蛋白Y-box基因在青虾不同组织中均有表达,其表达量在肝胰腺组织中最高,使用荧光定量PCR检测青虾冷休克蛋白Y-box基因在低温胁迫和恢复条件下在肝胰腺中的m RNA时空表达情况,结果显示,与对照组相比冷休克蛋白Y-box在肝胰腺中的表达量分别在低温和低氧胁迫3,6和12 h出现了显著上调,而在恢复刺激后其表达量与对照组差异不显著。此外,本实验对Y-box进行了原核表达,为进一步研究Y-box基因的功能奠定了基础。     

真蛸热休克蛋白90基因(HSP90)的克隆及表达

为深入了解真蛸热应激响应机制,实验以真蛸的应激蛋白为研究对象,借助同源扩增和cDNA末端快速扩增(rapid amplification of cDNA ends,RACE)技术获得真蛸热休克蛋白90(HSP90)的cDNA全序列(2 709 bp),它包含119 bp的5’非编码区(untranslated regions,UTR)、454 bp的3’UTR和2 136 bp的开放阅读框(opening reading frame,ORF),ORF共编码711个氨基酸,推算其分子量为81.5 ku,理论等电点为5.09。同源分析显示,所推导的氨基酸序列高度保守,并且含有HSP90家族特有的5个保守信号序列区域。实时荧光定量PCR分析表明,该基因在所有选取的组织中均有表达,肝组织中表达量最高。     

红螯光壳螯虾(Cherax quadricarinatus)酚氧化酶原基因的克隆与表达分析

为深入了解红螯光壳螯虾酚氧化酶原(proPO)基因的非特异性免疫机制,利用RACE技术从红螯光壳螯虾血细胞中克隆到酚氧化酶原基因cqproPO,cqproPO基因cDNA全长为2 962 bp,开放阅读框为1 998 bp,编码665个氨基酸,其结构中含有两个铜离子结合位点,预测分子量为75.86 ku;同源性比对结果显示,红螯光壳螯虾CqproPO与克氏原螯虾酚氧化酶原的同源性最高为79％,其次是淡水螯虾74％、挪威龙虾69％、美国龙虾67％等;进化分析发现CqproPO与克氏原鳌虾、淡水螯虾、挪威龙虾、美国龙虾等的酚氧化酶原亲缘关系最近;Realtime-PCR实验结果表明,CqproPO在血细胞中表达水平最高,其次是肠、触角腺、鳃等;在肝胰腺中有适量表达;WSSV感染后红螯光壳螯虾CqproPO mRNA在血细胞、肝胰腺和鳃组织中具有不同的时空表达趋势,但感染组和免疫后感染组mRNA表达量分别在感染后12h和24 h达到最大值,且在3种组织中2个感染组的CqproPO表达量为对照组的1.3 ～2.55倍,显著高于对照组(P＜0.05),之后cqproPO基因的转录水平明显下降.免疫后再受病毒感染的虾,CqproPO mRNA的表达量在3种组织中总体高于感染组,感染7d后的免疫保护率达到51.86％,表明免疫增强剂可使机体的抗病毒能力增强,对防御WSSV感染具有一定的免疫保护作用.     

红螯螯虾生物学特性的观察及与克氏螯虾的比较

本文描述了红螯螯虾的生物学特性,如分类地位、外形结构、栖息习性、消化器官与食性、呼吸器官及溶氧需求、蜕壳与生长、繁殖等,并与克氏螯虾进行了比较。     

Lecithin requirements of juvenile Australian red claw crayfish Cherax quadricarinatus

Australian red claw crayfish Cherax quadricarinatus is considered a popular crustacean species in several countries around the world because of its large size potential and resemblance to high‐priced American lobsters. However, little is currently known of the nutrient requirements and practical diet formulations for red claw. Lecithin has been shown to be required in the diets of several crustacean species, but there are no reports of dietary lecithin requirements for red claw. A 10‐week feeding trial was conducted in an effort to evaluate lecithin requirements for juvenile red claw. Juvenile red claw (mean individual weight of 1.6 ± 0.91 g) were individually stocked in a recirculating system at random into 80 plastic‐mesh culture units, each containing its own individual water line. There were 20 red claw per treatment (diet). Water was recirculated through biological and mechanical filters. Four semi‐purified diets were formulated to contain increasing percentages of commercial soya bean lecithin (0, 0.5, 1.0 and 2.0%). Diet ingredients included solvent‐extracted menhaden fish meal (FM), casein, dextrin, wheat flour, pellet binder, vitamin and mineral mix, cod liver oil, and corn oil. Semi‐purified diets were formulated to contain 40% protein using casein, menhaden FM, and wheat flour as protein sources. After 10 weeks, no significant differences (P > 0.05) were found in final weight, percentage weight gain and specific growth rate with average values of 13.0 g, 934%, and 3.14% day^?1, respectively. Percentage survival was high during the 10‐week period (100, 95, 100 and 95%) as only two individuals died during the study; one of these, because of an escape from the culture unit. There was also no significant differences (P > 0.05) in percentage moisture, protein, fat and ash in whole‐body red claw carcasses (wet‐weight basis) among any treatment (diet) and averaged 77.1, 12.6, 1.3 and 6.2%, respectively. Based upon the present study, these results indicate that a diet containing 5% cod liver oil and 1% corn oil, and having no supplemental lecithin, may be sufficient for growth and survival of juvenile red claw crayfish.     

Biochemical changes during vitellogenesis in the red claw crayfish,Cherax quadricarinatus (von Martens)

Redclaw crayfish (Cherax quadricarinatus) females at different stages of vitellogenesis were sampled to investigate variations in soluble protein, lipid and water content and in the amino acid and fatty acid composition of the hepatopancreas and ovaries. During vitellogenesis, the changes in the content of soluble proteins and lipids in the hepatopancreas and ovaries were dependent on both diet and the reserves from the hepatopancreas. However, in comparison to the ovary, the fluctuation of the amino acid composition in the hepatopancreas suggested that the protein resources mobilized from this organ was compensating for those gained from feeding. Changes in the fatty acid composition of the hepatopancreas showed limited compensatory function as for the quick accumulation of lipids in the ovaries. The proportional amounts of saturated fatty acids/mono‐unsaturated fatty acids/poly‐unsaturated fatty acids (PUFA) and the predominant fatty acids in both tissues indicated that the mobilization and utilization of fatty acids in freshwater species are different from that in marine species. Based on the redclaw's feeding habits, the commercial pellets used in this study may not be optimal, and a diet with less PUFA may suffice for its vitellogenesis and reduce the feeding costs.     

红螯螯虾胚胎发育过程中的生化成分变化

采用生物化学方法对红螯螯虾胚胎发育过程中主要生化成分(糖类、蛋白质及氨基酸、脂类及脂肪酸)进行了测定和分析,以期了解胚胎形态发生与营养需求的关系。结果显示：红螯螯虾胚胎发育过程中糖类总体含量很低。总蛋白与总氨基酸(TAA)的含量在胚胎发育过程中均呈下降趋势。在必需氨基酸(EAA)中亮氨酸（Leu）和精氨酸（Arg）的含量较高,而在非必需氨基酸中谷氨酸（Glu）和天冬氨酸（Asp）的含量较高。总脂在胚胎发育过程中的含量总体呈下降趋势,其中性脂和磷脂的主要脂肪酸均为C16:0、C18:0 、C18:1ω9和C18:3ω3。研究认为：在红螯螯虾胚胎发育过程中,糖类主要参与多种特异复合物的合成以及甲壳构建,这些特异复合物在细胞分化中起着重要的信号传递作用。蛋白质和脂类都是红螯螯虾胚胎发育过程中主要的结构和能源物质,蛋白质主要在早期被利用而脂类主要在晚期起作用。饱和脂肪酸（SFA）和单不饱和脂肪酸（MUFA）(C16:0,C18:0,C16:1ω7 和 C18:1ω9)主要作为能源物质被消耗而高不饱和脂肪酸（PUFA）(20:5ω3和22:6ω3)在胚胎发育过程中对于构建细胞膜以及调节中枢神经系统的发育起重要作用。     

红螯螯虾胚胎发育过程中的生化成分变化

采用生物化学方法对红螯螯虾胚胎发育过程中主要生化成分(糖类、蛋白质及氨基酸、脂类及脂肪酸)进行了测定和分析,以期了解胚胎形态发生与营养需求的关系.结果显示:红螯螯虾胚胎发育过程中糖类总体含量很低.总蛋白与总氨基酸(TAA)的含量在胚胎发育过程中均呈下降趋势.在必需氨基酸(EAA)中亮氨酸(Leu)和精氨酸(Arg)的含量较高,而在非必需氨基酸中谷氨酸(Glu)和天冬氨酸(Asp)的含量较高.总脂在胚胎发育过程中的含量总体呈下降趋势,其中性脂和磷脂的主要脂肪酸均为C16:0、C18:0 、C18:1ω9和C18:3ω3.研究认为:在红螯螯虾胚胎发育过程中,糖类主要参与多种特异复合物的合成以及甲壳构建,这些特异复合物在细胞分化中起着重要的信号传递作用.蛋白质和脂类都是红螯螯虾胚胎发育过程中主要的结构和能源物质,蛋白质主要在早期被利用而脂类主要在晚期起作用.饱和脂肪酸(SFA)和单不饱和脂肪酸(MUFA)(C16:0,C18:0,C16:1ω7 和 C18:1ω9)主要作为能源物质被消耗而高不饱和脂肪酸(PUFA)(20:5ω3和22:6ω3)在胚胎发育过程中对于构建细胞膜以及调节中枢神经系统的发育起重要作用.     

Effects of feeding practical diets containing various protein levels on growth, survival, body composition, and processing traits of Australian red claw crayfish (Cherax quadricarinatus) and on pond water quality

A 117‐day feeding trial was conducted in ponds with juvenile Australian red claw crayfish (Cherax quadricarinatus) to evaluate the effects on growth, survival, body composition, and processing traits when fed diets containing three different protein levels (22%, 32%, and 42%), and the effects of feeding these diets on pond water quality. Juvenile crayfish (mean weight of 4.6±2.2 g) were randomly stocked into nine 0.02‐ha ponds at a rate of 500 per pond (25 000 ha^?1), and each diet was fed to three ponds. There were two feedings per day, each consisting of one‐half of the total daily ration. At harvest, there were no significant differences (P>0.05) in the individual weight, percentage weight gain, or specific growth rate among treatments, which averaged 75.3 g, 1535%, and 2.38% day^?1 respectively. Red claw fed the 42% crude protein diet had significantly higher (P<0.05) feed conversion ratio (7.34) compared with crayfish fed diets containing 22% (5.18) or 32% (5.13) crude protein, and had significantly lower percentage survival (46.1%) compared with red claw fed diets with 22% (61.1%) or 32% (58.2%) protein. Total yield was significantly lower (P<0.05) in red claw fed the 42% protein diet (640 kg ha^?1) compared with red claw fed diets containing 22% (920 kg ha^?1) or 32% (904 kg ha^?1) protein. Mean total ammonia nitrogen (TAN) levels were significantly higher (P<0.05) in ponds with red claw fed the 42% protein diet (0.55 mg L^?1) compared with ponds with red claw fed diets containing 22% (0.32 mg L^?1) or 32% (0.38 mg L^?1) protein. Mean total nitrite concentrations in ponds with red claw fed the 42% protein diet was significantly higher (0.05 mg L^?1) compared with red claw fed diets containing 22% (0.01 mg L^?1) or 32% (0.02 mg L^?1) protein. These results indicate that a practical diet containing 22% (as fed basis) protein may be adequate for pond production of red claw when stocked at the density used in this study, and that a diet containing 42% protein adversely affected levels of TAN and nitrite, possibly reducing overall survival of red claw. Use of a diet with 22% protein may allow red claw producers to reduce diet costs and thereby increase profits.     

Effect of dietary protein level on the growth and survival of pre-adult freshwater crayfish Cherax quadricarinatus (von Martens) in monosex culture

Feeding trials with five levels of crude protein (CP) (22%, 27%, 33%, 39%, and 45%) and digestible energy ranging from 14.32 to 15.21 kJ g^?1 were carried out to determine optimum dietary protein for the growth and production of pre‐adult freshwater crayfish Cherax quadricarinatus. Triplicate groups of males (ω=23.1±0.58 g) and females (ω=21.8±0.33 g) were fed diets for 70 days in monosex culture. Optimal digestible protein (DP) was estimated for males by fitting mean weight gain to a quadratic model (y=27.86+0.839x–0.0216x², r²=0.945), and a linear model for females (y=32.84–0.0745x, r²=0.959). Optimal male growth occurred with 25.6% CP, equivalent to 19.4% DP. Male crayfish attained significantly higher final weights and absolute growth rate than females fed the 22%, 27%, and 33% CP diets. The results indicated that a 19.4% DP diet, with a P/E ratio of 11.4 mg kJ^?1, produces an optimum growth response for pre‐adult C. quadricarinatus under experimental conditions.     

Effect of different dietary protein and lipid levels on growth and survival of juvenile Australian redclaw crayfish, Cherax quadricarinatus (von Martens)

This study determined the effect of different dietary protein and lipid levels on growth and survival of juvenile redclaw Cherax quadricarinatus. Nine practical test diets were formulated to contain three crude protein (CP) levels [260, 310 and 360 g kg^?1, equivalent to 225, 260 and 296 g kg^?1 digestible protein (DP) respectively] at three crude lipid (CL) levels (40, 80 and 120 g kg^?1, equivalent to 38, 67 and 103 digestible lipids respectively), with digestible protein : digestible energy (DP : DE) ranging from 14.6 to 22.6 mg protein kJ g^?1. Three replicate groups of 15 crayfish (initial weight mean ± SD, 0.71 ± 0.13 g) per diet treatment were stocked in 40 L tanks, at 28 °C for 60 days. The highest mean weight, specific growth rate and biomass, with values of 7.0 g, 3.67% day^?1, and 370.2 g m^?2, respectively, were achieved by feeding a diet with P : L = 310 : 80 (P < 0.05). The treatments resulted in a survival rate of 80–91%, feed conversion ratio of 1.08–1.61 and protein efficiency ratio of 2.24–3.08. Results indicated that a diet containing 270 g kg^?1 DP (equivalent to 320 g kg^?1 CP), 75 g kg^?1 digestible lipid (DL) with a DP/DE of 18.4 mg protein kJ^?1, and 0.031 g protein per animal per day was optimum for juvenile C. quadricarinatus under the tested experimental conditions.