饲料中大豆磷脂对红螯光壳螯虾性腺发育期营养物质积累的影响

基础饲料中添加5种不同水平的大豆磷脂(SL):0%(Diet 1),1%(Diet 2),2%(Diet 3),4%(Diet 4)和6%(Diet 5)。饲喂初始体质量为(25.64±1.53)g的红螯光壳螯虾(Cherax quadricarinatus)雌虾8周,观察其卵巢发育期个体生长、性腺指数等参数的变化,分析卵巢和肝胰腺营养物质积累的快慢以及与营养物质积累相关基因脂肪酸结合蛋白(FABP)mRNA表达的差异。结果表明:饲料中不同水平的SL对卵巢发育期红螯光壳螯虾的成活率和相对增重率(WG)影响不显著(P>0.05),但饲喂≥2%(SL)饲料的雌虾性腺指数(GSI)显著升高(P<0.05),雌虾的肝胰腺指数(HSI)则随饲料中SL的增加而呈下降趋势(P>0.05);随着饲料中SL的增加,虾肝胰腺中脂滴的体积增大,数量增多,结构更加完整,质地更加均匀,卵巢中脂滴和卵黄颗粒的数量增多,体积增大;同时,饲料中SL的增加促进了FABP在肝胰腺和卵巢中的表达量。综上所述,饲料中的SL对红螯光壳螯虾雌虾卵巢发育具有促进作用,在培育红螯光壳螯虾雌虾亲体过程中,含6.5%鱼油的基础饲料至少补充2%的SL有利于雌虾卵巢的快速发育。本研究旨在为深入研究虾蟹生殖生理学和规模化红螯光壳螯虾早繁苗种生产等提供基础资料。     

饲料中非蛋白能源物质对红螯光壳螯虾幼虾生长、生理、生化指标的影响

在等蛋白质、等能量基础上,研究碳水化合物与脂类比例(CHO∶L)为10.75∶1、4.81∶1、2.66∶1、1.52∶1和0.87∶1的5组试验饲料对红螯光壳螯虾[初始体质量(1.72±0.01)g]相关生长、生理、生化指标的影响。8周试验结果表明,CHO∶L比例为2.66∶1时,红螯光壳螯虾的增重率、特定生长率和饲料利用率达到最高。高比例的CHO∶L(10.75∶1)和低比例的CHO∶L(0.87∶1)都会显著地抑制(P<0.05)红螯光壳螯虾的生长和饲料的利用。饲料脂肪水平为40～145 g/kg时,虾的脂肪酶和碱性磷酸酶活力显著升高(P<0.05),己糖激酶和丙酮酸激酶活力则呈显著降低趋势(P<0.05)。CHO∶L对虾胃蛋白酶活力影响显著(P<0.01),CHO∶L为2.66∶1和1.52∶1表现出比较高的活力,显著高于(P<0.05)其它试验组。碳水化合物为156.3～360.4 g/kg范围内,虾淀粉酶活力随饲料中碳水化合物的升高而显著升高(P<0.01)。红螯光壳螯虾增重率分别与饲料中碳水化合物和脂肪水平进行二次回归分析得出,红螯光壳螯虾对配合饲料中碳水化合物和脂肪的最适需求量分别为268.28和120.22 g/kg,相对应的CHO∶L为2.20∶1,且红螯光壳螯虾对碳水化合物的利用能力要高于对脂肪的利用。     

不同脂肪源对红螯光壳螯虾幼虾生长、消化酶活性及其肌肉生化组成的影响

通过在人工配合饲料中分别添加5%的花生油、猪油、鱼油和豆油作为主要脂肪源,以商业饲料为对照,进行8周饲喂实验,研究不同脂肪源对红螯光壳螯虾(Cherax quadricarinatus)幼虾生长、消化酶活性及其肌肉生化组成的影响。结果显示,不同脂肪源对红螯光壳螯虾幼虾的体长增长率和特定增长率影响不显著(P0.05);但增重率各组间存在显著性差异(P0.05),以豆油组最高,达到2332.93%,花生油组最低,为1839.50%;豆油组肝胰腺指数显著高于其他各组(P0.05),为0.75%。幼虾存活率以豆油组最高(P0.05),达到83.3%,鱼油组较低,仅为56.7%。不同脂肪源饲喂组的肝胰腺胃蛋白酶活力无显著差异(P0.05);脂肪酶活力花生油组显著高于其他实验组(P0.05),为1177.23U/g(prot);淀粉酶活力各组间差异显著(P0.05),由高到低依次为鱼油组、对照组、豆油组、猪油组、花生油组;纤维素酶活力以花生油组较高,为61.14U/(gprot()P0.05)。幼虾腹部肌肉中各种脂肪酸的含量明显受到饲料中脂肪酸种类和含量的影响,饱和脂肪酸含量猪油组显著高于其他组(P0.05);单不饱和脂肪酸以花生油组含量最高,豆油组最低;多不饱和脂肪酸则以豆油组含量最高(P0.05)。在各实验组中,豆油组红螯光壳螯虾幼体具有最高的体质量增长率和存活率,较高的肝胰腺指数和肝胰腺消化酶活力,豆油组幼虾腹部肌肉中多不饱和脂肪酸尤其是亚油酸和亚麻酸含量也较高。因此,以豆油作为主要脂肪源能够满足红螯光壳螯虾幼体的生长需要,获得较好的饲养效果,并降低饲料成本。     

不同生长时期红螯螯虾肌肉和肝胰腺内氨基酸和脂肪酸变化分析

为了研究不同生长时期红螯螯虾(Cherax quadricarinatus)肌肉和肝胰腺内脂肪酸和氨基酸的动态变化,选取90 d (第二性征未发育)、140 d (第二性征发育成熟)和180 d (性腺发育成熟)的红螯螯虾,分别测定其肌肉和肝胰腺内氨基酸和脂肪酸的含量。结果显示,90 d红螯螯虾肌肉中的必需氨基酸总量显著高于其他2个时期(P<0.05);而红螯螯虾肝胰腺内必需氨基酸总量和非必需氨基酸总量均随着日龄增加而逐渐下降。比较不同生长时期肌肉中脂肪酸含量发现,随着日龄增长,饱和脂肪酸(SFA)含量呈下降趋势;而单不饱和脂肪酸(MUFA)含量呈先上升后下降的变化趋势,140 d含量最高[(25.69±0.42)%];多不饱和脂肪酸(PUFA)含量呈上升的变化趋势。肝胰腺中SFA含量在 3个生长时期内无显著性差异(P>0.05),MUFA含量呈先上升后下降的变化趋势,140 d含量最高[(37.44±0.59)%],而PUFA含量变化与MUFA正好相反。营养价值评估结果显示,180 d (可上市销售)的红螯螯虾肌肉内必需氨基酸指数(49.96%)高于联合国粮农组织/世界卫生组织(FAO/WHO)评分模式(31.50%)和全鸡蛋蛋白质模式(43.10%),是较为理想的优质食物蛋白源。由上可知,在不同生长时期,肌肉和肝胰腺中的氨基酸、脂肪酸的主要消耗不同,研究结果对了解不同生长时期红螯螯虾营养需求、开发红螯螯虾配合饲料提供了参考依据。     

红螯螯虾不同体色个体间营养差异分析

为了探究红螯螯虾（Cherax quadricarinatus）体色与其营养成分之间的连锁关系,测定分析棕黄色[体质量为（51.8±4.2）g]、草绿色[体质量为（52.9±3.4）g]和天蓝色[体质量为（48.5±10.9）g]三种体色个体肌肉的营养组分。结果显示,草绿色个体腹部肌肉的粗蛋白含量显著高于棕黄色和天蓝色个体（P<0.05）。棕黄色、草绿色、天蓝色三种体色个体的必需氨基酸与总氨基酸比值分别为38.83%、41.52%和34.61%;必需氨基酸与非必需氨基酸的比值均超过60%;草绿色个体显著高于棕黄色和天蓝色个体（P<0.05）,属于优质蛋白;呈味氨基酸与总氨基酸比值分别为46.87%、41.94%和53.41%,天蓝色个体显著高于草绿色和棕黄色个体（P<0.05）,表明天蓝色个体的肉质风味更加鲜美。三种体色的红螯螯虾之间总饱和脂肪酸差异显著,棕黄色个体显著高于草绿色和天蓝色个体（P<0.05）;饱和脂肪酸主要以棕榈油酸（C16:0）为主,棕黄色个体中C16:0的含量高于草绿色和天蓝色个体,并与草绿色个体形成显著性差异（P<0.05）。在多不饱...     

切除单侧眼柄对中华绒螯蟹蜕壳、生长、成熟的影响

室内条件下,采用烫灼挤压法切除中华绒螯蟹的单侧眼柄,结果表明,对青春期蜕壳前的蟹,切除任意一侧眼柄,都能有效地促进蜕壳、生长及性腺的早期发育,但无助于诱导卵巢的成熟、产卵.试验还表明,切除单侧眼柄不会导致高死亡率,且对蟹的交配、怀卵、幼体的孵化培育无不良影响。     

水草和螺蛳对克氏原螯虾性腺发育的影响

本文对水草、螺蛳、水草+螺蛳三种投喂方式对克氏原螯虾性腺发育的影响进行了研究。结果显示,水草+螺蛳组的克氏原螯虾(雌虾体长9.5cm)的抱卵量826粒/尾,显著高于水草组、螺蛳组(P<0.05),成熟度5.95%、抱卵指数19.20粒/g、亲虾成活率89.66%、抱卵率79.31%,结果均高于投喂水草组或螺蛳组。显然,植物性和动物性饵料混合喂养更有利于促进克氏原螯虾雌虾性腺发育及提高亲虾的成活率。     

不同温度对红螯螯虾虾苗生长的影响

以体质量0.02 g红螯螯虾为研究对象,开展了不同温度(22、25、28、31、34℃)对红螯螯虾虾苗生长影响的试验。试验结果表明随着温度的升高,红螯螯虾虾苗的成活率不断降低,22℃时成活率最高,为48.21%±8.50%,34℃时成活率最低,为21.55%±3.09%,31℃时虾苗特定生长率最高,为7.50%±0.41%。     

温度和氨氮对不同规格红螯螯虾耗氧率与排氨率的影响

为了揭示温度和氨氮浓度对红螯螯虾(Cherax quadricarinatus)耗氧率(RO)与排氨率(RN)的影响,采用单因素实验设计和密闭流水法分析了不同温度和氨氮浓度条件下,3种规格红螯螯虾(小规格为S组、中规格为M组和大规格为L组) RO和RN的变化规律。结果显示,温度对红螯螯虾RO和RN影响显著(P<0.05),在温度为15℃~35℃时,RO随温度升高而增加,35℃时,红螯螯虾的RO达到最大值;其中,S组的RO为0.777 mg/(g·h),显著大于其他2个组(P<0.05)。红螯螯虾RN随温度升高呈先上升后下降的趋势,30℃时达到最大值;其中,S组的RN为0.061 mg/(g·h),显著大于其他2个组(P<0.05)。温度对红螯螯虾的氧氮比(O/N)值有显著影响(P<0.05)。氨氮浓度对红螯螯虾的RO和RN影响显著(P<0.05),在氨氮浓度为0~16 mg/L时,二者均随氨氮浓度的上升呈先上升后下降的趋势;当氨氮浓度为8 mg/L时,红螯螯虾的RO达到最大值,S、M和L组RO值分别为0.663、0.332和0.195 mg/(g·h);当氨氮浓度为12 mg/L时,RN达到最大值,S、M和L组RN值分别为0.123、0.049和0.034 mg/(g·h),且显著高于其他处理组(P<0.05)。氨氮浓度对3种规格红螯螯虾的O/N值均有显著影响(P<0.05)。不同温度条件下,红螯螯虾RO和RN的Q10变化范围分别为1.102~3.361和0.346~3.417,且分别在25℃~30℃和30℃~35℃时达到最小。     

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.     

Effect of Acclimation Temperature and TemperatureChanges on Molting and Survival of Ablated and Non-ablated Crayfish Orconectes virihs

The effect of acclimation temperatures for producing soft-shell crayfish, and changes upward from acclimation temperature, were evaluated in crayfish Orconectes virilis subjected to either hemilateral or bilateral eyestalk ablation. In each experiment, 60 juvenile O. virilis were reared for 21 d at temperatures ranging from 20 C to 30 C. Equal numbers of male and female crayfish ranging in weight from 5.0 g to 16.0 g were used in each study. In the first three experiments, crayfish were acclimated and maintained at a constant temperature. In the remaining experiments, crayfish were subjected to a step-wise increase in temperature of either 5 or 10 C. Molting was highest in crayfish acclimated to 25 C prior to experimentation at 25 C (experiment 2) and in crayfish acclimated to 14 C prior to experimentation at 20 C (experiment 4). Total molts during these two experiments were 51 (85.0%) and 56 (93.3%), respectively. Bilateral eyestalk ablation resulted in significant reductions in the intermolt period in experiments 1 (acclimation temperature 20 C, experimental temperature 20 C), 3 (acclimation temperature 30 C, experimental temperature 30 C) and 6 (acclimation temperature 15 C, experimental temperature 25 C), while hemilateral eyestalk ablation had no effect on molting when compared to control (non-ablated) crayfish. Bilateral eyestalk ablation resulted in a significant reduction in survival in experiments 5 (acclimation temperature 20 C, experimental temperature 25 C) and 6 (acclimation temperature 15 C, experimental temperature 25 C). Significantly more females molted in one experiment (acclimation temperature 20 C, experimental temperature 25 C), but not in any others. The results of these studies demonstrate that bilateral eyestalk ablation and water temperatures are effective methods of inducing molting in crayfish.     

Unilateral eyestalk ablation improves molting frequency and reproduction in Macrobrachium amazonicum females

This study has investigated the influence of unilateral eyestalk ablation on the survival, molting frequency, fecundity, reproductive output, and fertility of M. amazonicum females. We divided 96 females into with ablation and without ablation groups with six replicates. Water quality, growth, survival rate, intermolt period, molts number, and ovigerous females over a 90-day period were monitored. Unilateral eyestalk ablation reduced intermolt period by 30% and increased molts number, ovigerous females, and reproductive output. Our findings indicate that unilateral eyestalk ablation positively influences reproduction in M. amazonicum and is appropriate for females of this species having a body length of ≥ 70 mm.     

Indoor Spawning and Egg Development of the Red Claw Crayfish Cherax quadricarinatus

In 1991 and 1992, indoor hatchery experiments were conducted with red claw crayfish Cherax quadricarinatus a large tropical crayfish from northern Australia. Six fiberglass tanks were stocked with 40–190 g mature red claw crayfish at 10/m² and a 1:2 male to female ratio. Water temperature was maintained at 28 ± 1 C. Photoperiod was provided by natural daylight in 1991 and controlled lighting in 1992.
With natural lighting, monthly spawning rates (% of females spawning/month) ranged from an average of 15% during months with less than 12 h of light per day to over 35% in months with more than 12 h of light per day. Peak spawning occurred from May to July, too late to obtain juveniles for spring stocking. During the second year, photoperiod was increased from 10L:14D in December to 14L:10D in February and maintained at this level until July. Red claw spawning increased with increasing day length. Peak spawning rates of 50% per month and higher occurred from March to May. Spawning rates decreased dramatically after May, even though photoperiod and temperature remained constant. Eggs per spawn increased with an increase in female size. Fecundity average 7.3 juveniles per gram of body weight for females from 40 to 190 g, regardless of size. Egg color was useful for predicting stage of development.     

Alterations of pattern in immune response and vitellogenesis during induced ovarian development by unilateral and bilateral ablation in Litopenaeus vannamei

Eyestalk ablation is often used to induce ovarian development in female shrimp, but it possibly alters the animal’s hormonal balance, and affects its physiology and immune system. Therefore, this study was conducted to investigate alterations to the immune response during induced ovarian development by unilateral and bilateral ablation in Litopenaeus vannamei. Sub-adult female shrimp could be induced to undergo ovarian development over a 20-day period by both unilateral and bilateral eyestalk ablation; however, ovaries did not reach the final mature stage in which cortical rods are observed. In terms of immune response, total hemocyte count decreased significantly at 4 h after unilateral ablation, and at 20 days after bilateral ablation. After 20 days, expression in the hemocytes of the immune-related genes prophenoloxidase and peroxinectin in bilaterally ablated shrimp increased significantly, whereas that for penaeidin, for lipopolysaccharide- and β-glucan-binding protein, and for transglutaminase mRNA expression did not change significantly. The results of this study suggest that, while eyestalk ablation induces ovarian development, it may also bring about a hormonal imbalance that leads to immune-related genes being activated.     

Preliminary Pond Production of the Red Claw Crayfish,Cherax Quadricarinatus,in the Central United States

The Australian red claw crayfish, Cherax quadricarinatus, has recently attracted attention as a potential candidate for aquaculture in the United States. This paper reports the first experimental yield trials of the red claw crayfish in the central United States. Two earthen ponds (0.30 and 0.31 ha) were stocked with juvenile red claw crayfish at either 3.5 or 7.0 crayfish/m². Crayfish were fed a diet of pelletized shrimp ration supplemented with corn silage. When stocked at 3.5/m₂, red claw crayfish grew from an average weight of 3.0 g to 27.0 g in 97 days; average survival was 79.6%; net yield was 653 kg/ha; and feed conversion was 1.4. When stocked at 7.0/m² red claw crayfish grew from an average weight of 4.1 g to 19.7 g in 99 days; average survival was 70.0%; net yield was 678.9 kg/ha; and feed conversion was 1.8. During the yield trial, temperatures were optimal for growth (25 to 33°C) 60% of the time. These findings are sufficiently encouraging to warrant further trials in locations with a longer growing season, in order to evaluate more fully the potential of the red claw crayfish as a candidate for aquaculture in the United States.     

切除单侧眼柄对不同体质量养殖斑节对虾卵巢发育及产卵的影响

在盐度31、水温28~29℃下,进行了4次卵巢诱导试验,研究了切除单侧眼柄对体质量56.4~105.4g的养殖斑节对虾卵巢发育和产卵的影响及卵巢发育、产卵与体质量之间的关系。结果显示,切除单侧眼柄诱导养殖斑节对虾卵巢发育成熟,成功率(y)与雌虾体质量(x)显著相关(P0.05),雌虾体质量越小,诱导成功率越低,回归方程为:y=133.92lnx-535.63(r2=0.9297)。养殖斑节对虾眼柄手术后,最快10d产卵,部分雌虾能重复产卵。产卵量(y)与雌虾体质量(x)显著正相关(P0.05),雌虾体质量越小,产卵量越小,回归方程为:y=122.22lnx-472.2(r2=0.9738)。雌虾累积死亡率、卵子直径和卵子孵化率与雌虾体质量无显著相关性(P0.05)。建议用体质量80g以上的养殖斑节对虾雌虾作为亲虾。     

泛甲壳动物激素调控卵黄蛋白原合成的分子机制研究进展

昆虫和甲壳动物由于进化上亲缘关系近统称泛甲壳动物，大多数泛甲壳动物卵黄蛋白原为雌性特异性蛋白，是卵母细胞中储存的卵黄蛋白的前体，可为胚胎发育提供营养和能量，是决定繁殖性能的关键因素，在生殖发育起着十分重要作用。业已证明，调控泛甲壳动物卵黄蛋白原的主要激素有保幼激素、蜕皮激素、神经肽和胰岛素样肽等。昆虫保幼激素和蜕皮激素发挥促进作用，而神经肽和胰岛素样肽对卵黄蛋白原合成的调控作用因种类不同存在不同。除了以上激素，甲壳动物特有的眼柄高血糖激素家族以及促雄腺激素对卵黄蛋白原的合成起负调控作用。本文总结了近年来泛甲壳动物卵黄蛋白原合成的调控机制的相关研究，概括并比较了调控昆虫与甲壳动物的卵黄蛋白原合成的主要激素及调控机制，为养殖虾蟹甲壳动物生殖调控相关研究提供参考。     

克氏原螯虾PcCyc基因克隆及其对光周期的表达响应

利用cDNA末端快速扩增技术克隆克氏原螯虾生物钟核心基因之一的周期循环蛋白(CYC)基因(命名为PcCyc基因)的部分cDNA序列。取雄性克氏原螯虾(体长9~11 cm)在全光谱日光灯不同光周期下(12L∶12D、0L∶24D和0L∶72D)培养结束后,于7:00、11:00、15:00、19:00、23:00、翌日03:00、7:00取样,采用实时荧光定量PCR检测PcCyc基因在脑、眼柄和肝胰腺中的mRNA表达水平。试验结果显示,PcCyc基因完整的开放阅读框共2010个碱基,可编码669个氨基酸。氨基酸序列与红螯螯虾CYC蛋白相似性为94%;蛋白分子量约73.963 ku,理论等电点6.56,为疏水性非分泌蛋白,无信号肽;结构域分析结果显示,PcCYC蛋白含有BHLH/PAS结构域,包括1个HLH结构域和2个PAS结构域;二级结构包含11个α-螺旋、12个β-折叠和24个无规则卷曲。不论光照如何改变,PcCyc基因在眼柄和肝胰腺中都基本遵循着24 h的节律振荡,而在大脑中的节律振荡随光照周期改变被打破。试验结果将为进一步探究克氏原螯虾Cyc基因在其昼夜生物钟节律中发挥的作用及机制奠定基础。     

Production of Australian Red Claw Cherax quudricarinatus in Polyculture with Nile Tilapia Oreochromis niloticus

Abstract— Australian red claw crayfish Cherax quudricarinurus were cultured in monoculture and in polyculture with male Nile tilapia Oreochromis niloricus in nine 0.02-ha earthen ponds. Red claw with an average weight of 7 g were stocked in all ponds at 2/ m². Tilapia with an average weight of 19 g were stocked in six ponds at either 0.5 or 1/m². A sinking 32% fish diet and dry alfalfa hay was provided to all ponds as feed and forage. After 135 d, red claw survival was similar (19–24%) in all treatments while average weights were 76 g in monoculture and 48 g in both polyculture treatments. Lower than expected survival was a result of stress caused by poor shipping conditions. Tilapia survivals were similar (84–90%) in all ponds and had an average weight of 403 g when stocked at 1/m² and 444 g when stocked at 0.5/m². Red claws are generally considered non-aggressive, especially when compared with other clawed crustaceans. This lack of aggressiveness is suggested as a cause for decreased growth in polyculture with tilapia.