刺参与日本对虾池塘高效混养技术

刺参夏眠期间正是日本对虾生长的大好季节,将刺参和日本对虾混养,在7月份放养虾苗,严格控制养殖池内的杂鱼虾等,就可大大提高日本对虾和刺参的成活率,利用此二者在饵料和空间的互补性,可显著提高虾池的利用率和经济效益。     

免疫低聚糖对中国对虾的免疫效用研究

为证实免疫低聚糖(D-氨基葡聚寡糖,D-Amino-Oligosacellaride)对中国对虾(Penaeus chinensis)的免疫增强作用,进行了不同剂量的注射及饲料投喂实验.结果表明,注射免疫低聚糖可显著增强中国对虾酸性磷酸酶和溶茵酶等血清免疫指标,添加于饲料中投喂可以提高对虾成活率,降低饵料系数,且饲料中添加量≥3‰的效果较佳.     

血清素在斑节对虾催熟过程中的作用

在斑节对虾 Penaeus monodon 催熟催产过程中,常用的方法是切除眼柄.此实验选用体重为149.5±25.8 g的自然海区斑节对虾,比较了注射血清素(serotonin)和切除眼柄2种处理方法对斑节对虾的催熟效果.血清素注射剂量为50 μg·g -1体重,在实验的第1、5和10天注射,共计3次.结果显示,2种方法处理后,受试亲虾性腺指数(gonad somatic index,GSI)显著高于对照组,同时注射血清素的实验组亲虾的存活率及其卵子孵化率显著高于切除眼柄的实验组.实验结果表明,血清素对斑节对虾催熟有促进作用,但发挥效应所需要的时间较长;从切除眼柄手术到第一次产卵所需时间较短,但是手术后的亲虾死亡率较高.     

中国对虾皮下及造血组织坏死杆状病毒（HHNBV）DNA探针的制备及应用

ＨＨＮＢＶ是１９９３年中国对虾暴发性流行病的主要病原之一。从纯化的病毒中提取ＤＮＡ，用ＥｃｏＲｌ限制性内切酶酶切成ＤＮＡ片段，与ＰＵＣ１８体外重组，建立ＨＨＮＢＶＤＮＡ文库。从中筛选出三个重组质粒（５＃、８＃、９＃），它们所插入的片段大小分别为：２．８Ｋｂ、０．８Ｋｂ、１．６Ｋｂ，它们分别用光敏生物素标记成探针，与感染ＨＨＮＢＶ的病虾ＤＮＡ呈阳性反应，以此利用制备的ＨＨＮＢＶＤＮＡ探针的高敏感性和特异性来检测虾病。     

斑节对虾基因组微卫星分离及其序列特征研究

采用(CA)_(12)(AG)_(12)及(TA)_(16)生物素标记探针及磁珠富集法构建了斑节对虾Penaeus monodon基因组微卫星富集文库。随机挑选254个克隆进行PCR筛选,得到51个候选克隆(20．1%)。其中,32个克隆来源于CA-文库,另19个克隆来源于AG-文库。测序发现48个克隆含有微卫星重复单元,通过序列比对,最终获得40个具有特异微卫星序列的阳性克隆。微卫星(GA/CT)_N及(CA/GT)_n 2碱基重复序列分别占所有分离的微卫星数目的20．7%及60．4%。此外,还检测到其它多种微卫星重复类型,如(AT)_n、(GC)_n、(TGG)_n、(AAG)_n、(AAT)_n、(GAA)_n、(GTGC)_n、(GCGT)_n、(GGTTA)_n、(GTGCGT)_n,占检测到的微卫星数目的18．9%。获得的微卫星序列中属于完全型序列的有76条(68．5%),不完全型序列的有22条(19．8%),另有13条属于复合型序列(11．7%)。微卫星(GT/CA)_n 2碱基重复次数(3～52次)要远大于(GA/CT)_n 2碱基次数(3～27次)。获得的微卫星序列长度大小范围为129～601 bp,平均为286 bp。研究为进一步开展斑节对虾分子育种及资源评价分析提供了基础资料。     

Genetic parameters for spawning and growth traits in the Pacific white shrimp (Penaeus (Litopenaeus) vannamei)

The aim of this study was to estimate genetic parameters for spawning traits and growth traits in a breeding line of Pacific white shrimp, Penaeus (Litopenaeus) vannamei, selected for growth and survival. Traits studied were number of eggs (NE) and number of nauplii (NN) and female body weight at insemination (FWI) and body weight at 130 days of age (BW130). Genetic parameters were estimated using a multivariate animal model. Heritability for NE and NN were estimated as 0.13 ± 0.04 and 0.03 ± 0.04 respectively. The contribution to NN total variation due to ‘factors associated with male’ effect was estimated as 0.47 ± 0.07. In the cases of FWI and BW130, heritability was estimated as 0.44 ± 0.08 and 0.19 ± 0.03 respectively. Genetic correlation between FWI and NE was estimated as 0.49 ± 0.15, between FWI and NN as 0.54 ± 0.39 and between NE and NN as 0.27 ± 0.41, whereas the genetic correlations of FWI, NE and NN with BW130 were 0.30 ± 0.13, ?0.21 ± 0.19 and ?0.25 ± 0.38 respectively. Although it is important to perform more studies on this issue, our results found no evidence of a genetic antagonistic effect between female reproductive traits and body weight at harvesting (130 days of age) in P. vannamei.     

碳酸盐碱度胁迫下凡纳滨对虾基因的差异表达

以广盐性养殖的凡纳滨对虾(Litopenaeus vannamei)为研究对象,采用抑制消减杂交(suppression subtractive hybridization,SSH)和定量PCR的方法,研究其在高碳酸盐碱度胁迫下转录组水平的基因表达差异,以期从基因组水平研究对虾对高碳酸盐碱度胁迫的适应机制。结果表明,以高碳酸盐碱度(20 mmol/L)胁迫第4天凡纳滨对虾鳃组织和低碱度(2 mmol/L)对照组鳃组织为材料,通过斑点杂交筛选,发现鳃组织中有158个克隆子表达上调,291个克隆子表达下调。挑选150个高表达差异的克隆子进行测序,获得100个序列,其中50个得到成功注释。经过GO分析,这些注释的差异基因主要分为8大类群,其中碳酸酐酶基因(CA)、Na+-K+-ATPase基因(NKA-α)等与离子调控相关的基因表达量上调,而溶菌酶基因等与先天免疫相关的基因表达量下调,这些结果表明高碳酸盐碱度胁迫下,凡纳滨对虾以增加离子调控的方式进行酸碱平衡的调控,同时其免疫功能受到抑制。此外,还对CA和NKA-α两个基因在鳃和触角腺中的时空表达规律进行了研究,发现高碳酸盐碱度胁迫9 d过程中,两个基因在鳃组织中的表达均呈现先高表达后回落的现象,而在触角腺中NKA-α基因则一直维持较高表达水平,说明CA和NKA-α基因在凡纳滨对虾高碳酸盐碱度适应离子调控中起着关键作用,同时还发现除了鳃组织之外,触角腺同样参与了调控。本研究从转录水平初步筛选了高碳酸盐碱度胁迫下凡纳滨对虾的表达差异基因,探索了凡纳滨对虾的耐盐碱机制,对培育适于盐碱地水产养殖的优良品种有着重要的意义。     

凡纳滨对虾α-淀粉酶基因的SNPs检测

利用PCR产物直接测序法,对凡纳滨对虾SPR和SPF 2个品种共40个样本的α-淀粉酶基因(GenBank序列号:AJ133526)的单核苷酸多态性(SNP)进行了研究。找到9个SNPs,分别为:C127T、A138G、T951C、T1083C、C1457T、A2147C、A2226G、A2297G和T2306C。其中1个SNP是第4外显子中的错义突变,2个SNPs分别是第5和第7外显子中的同义突变,6个SNPs是内含子中的突变。这些结果可为凡纳滨对虾的研究提供遗传学依据。     

A3α-肽聚糖对凡纳滨对虾磷酸酶及细胞内酚氧化酶活性的影响

为考察A3α-肽聚糖(Peptidoglycan,PG)在凡纳滨对虾(Litopenaeus vannamei)的养成期对其免疫酶活性的影响,设立了连续投喂、间隔投喂和浸浴共6个实验组。连续投喂组中PG的添加量分别为0(对照)、0.01%、0.05%和0.1%;间隔投喂组中PG的添加量为0.05%;浸浴组的给予方式为50mg/ml,3h/次/15d。分别在30、60和90d时,测定对虾体内及血浆上清中的酸性磷酸酶(Acid phosphatase,ACP)和碱性磷酸酶(Alkaline phosphatase,AKP)活性,另外,在60d和90d时,测定对虾血细胞内酚氧化酶(Phenoloxidase,PO)活性。结果表明,30d时,PG浸浴组、0.05%和0.1%PG添加量组的ACP活性,各PG作用组的AKP活性均较对照组高,且0.1%PG添加量组的ACP活性及0.05%和0.1%PG添加量组的AKP活性较对照组增长显著(P<0.05);60d时,除间隔投喂组外,各组的ACP活性均较对照组高,且0.1%PG添加量组增长显著(P<0.05),而各实验组的AKP活性均较对照组高,且0.05%PG添加量组增长显著(P<0.05),除浸浴组外,各组血细胞内PO活性均有提高;90d时,浸浴组和间隔组的ACP和AKP活性均较对照组高,且间隔投喂组的AKP活性增长显著(P<0.05);间隔投喂组的血细胞内PO活性较对照组有增长。实验证明,PG能激活对虾的免疫力,并可作为免疫增强剂应用于对虾的生产。     

Observations on growth, sexual maturity and spawning performance of pond-reared Penaeus merguiensis

This 12‐month preliminary study investigated the development of sexual characters, primary sexual maturity, ovarian maturity and spawning performance of pond‐reared Penaeus merguiensis in relation to culture conditions in south‐east Queensland, Australia. Post‐larvae of P. merguiensis were produced and cultured in two 60‐m³ tanks during the first 14 weeks. Before winter, they were harvested and stocked in three different overwintering facilities: a 200‐m² covered pond, two 60‐m³ outdoor tanks and a 15‐m³ indoor, recirculated tank at a stocking density of 10 individuals m^?2. The development of sexual characters was found to be similar to that reported previously in wild P. merguiensis. Males matured at younger ages and smaller sizes than females. Overall, the average size at primary sexual maturity of pond‐reared P. merguiensis was 23.1 mm carapace length for males (possessed spermatophores) and 29.3 mm for females (being impregnated). Water temperature and the availability of natural food strongly influenced prawn growth, maturity rate and their subsequent spawning performance. Growth, maturity rate and spawning performance of prawns in the covered pond were significantly higher than in the other overwintering facilities. Prawns started mating at 6–7 months, reached full ovarian maturation and spawned as early as about 8 (peaked at 9–11) months from hatching, producing high fecundity and viable larvae. There was a strong relationship (P < 0.001) between prawn size and fecundity. The results of this study suggest a potential for using pond‐reared broodstock P. merguiensis for hatchery production and for domestication or selective breeding programmes.