Spermatophore cryopreservation and artificial insemination of black tiger shrimp, Penaeus monodon (Fabricius)

To develop an appropriate cryopreservation protocol for spermatophores of black tiger shrimp, Penaeus monodon, three cryoprotectants (dimethyl sulphoxide (DMSO), methanol (MeOH) and ethylene glycol (EG)) at two concentrations (5% and 10%) were examined. Artificial implantation of spermatophores was also carried out to assess the fertilizing ability of fresh and post‐thaw spermatophores. Spermatophores were collected during consecutive regenerations (15‐day intervals) and assessed for qualitative and quantitative changes and also for fertilizing ability by implantation. The mean fertilization rate for artificial insemination using post‐thaw spermatophore was 79.9±3.7%, lower than the fertilization rates observed for artificial implantation using fresh spermatophore and natural mating. Mean hatch rates for fresh spermatophore, frozen‐thawed spermatophore and natural mating were 88.8±0.6%, 87.8±0.4% and 88.3±0.5%, respectively; and there was no difference among the three groups. The mean fertilization rate of spermatophores collected during the first stripping was higher (90.6±0.6) than during the second stripping (85.7±2.6), but the mean hatch rate was not different between the two strippings. The highest mean sperm viability (79.7±0.4%) was obtained from DMSO (5%), with no survival observed in the 10% MeOH treatment. Spermatophore weight, total sperm count and percentage of abnormal sperm were not different between spermatophores collected at the first and second stripping. This is the first study to report high fertilization and hatch rates from cryopreserved spermatophore using artificial implantation of spermatophore before spawning.     

Expression of a recombinant crustacean hyperglycemic hormone of the kuruma prawn Penaeus japonicus in methylotrophic yeast Pichia pastoris

ABSTRACT:   Until now, six crustacean hyperglycemic hormones (CHH) designated Pej-SGP-I, -II, -III, -V, -VI and -VII have been characterized in the kuruma prawn Penaeus japonicus . All CHH consist of 72 amino acid residues and have an amidated carboxyl (C)-terminus. In the present study, we expressed Pej-SGP-III in methylotrophic yeast Pichia pastoris in order to obtain a large quantity of recombinant CHH possessing biological activity. A cDNA encoding Pej-SGP-III that had been previously cloned was processed by polymerase chain reaction (PCR) and the resulting product was ligated into an expression vector. Pichia pastoris was transformed with this vector after which a recombinant Pej-SGP-III was expressed having an additional amino acid residue (glycine) at the C-terminus (rPej-SGP-III-Gly), a form considered to be a putative precursor of this hormone. rPej-SGP-III-Gly secreted into the culture medium was purified by reversed-phase HPLC, and amidated using a peptidylglycine alpha-amidating enzyme. The amidated rPej-SGP-III (rPej-SGP-III) showed hyperglycemic activity in in vivo bioassay almost comparable to that of the natural Pej-SGP-III. rPej-SGP-III thus obtained will be a useful tool not only for its physiological study but also for the determination of its 3-D structure.     

南美白对虾池塘养殖水质智能控制系统设计与开发

南美白对虾池塘养殖对于水体质量有着较为严苛的要求,残饵及生物排泄物的增多会大量消耗水体溶解氧,进而打破水体动态平衡,导致NH₃-N、NO₂-N等毒害成分升高,引起南美白对虾病害甚至导致死亡,造成养殖户经济损失并且阻碍了南美白对虾养殖产业的健康发展。针对上述问题,笔者研究并开发一种能够实时监测养殖池塘水质,并能根据南美白对虾生长习性智能控制增氧机、水泵等养殖设备的南美白对虾池塘养殖水质智能控制系统。系统具有控制智能、运行可靠、集成度高等特点,在养殖生产试验中取得了良好效果,保证了南美白对虾养殖过程的安全,有效降低了南美白对虾池塘养殖风险。     

凡纳对虾小触角表面结构的显微观察

应用光学显微镜和扫描电子显微镜观察了凡纳对虾小触角的表面结构。结果发现：小触角上广泛着生着700根以上的羽状刚毛；外鞭片状隆起腹面平均约具280根无顶孔但有细分支的化感刚毛，这类有分支的化感刚毛在其它十足目种类上未见有报道，而且其刚毛数也较其它种类少；内侧附肢前端与柄节上直立的羽状刚毛毛簇顶相接触，推测该结构可能对相邻的复眼具某种作用。     

保幼激素拮抗物KK-42对南美白对虾Y-器官发育的影响

采用1.95×10-4mol.L-1的KK-42溶液浸润处理南美白对虾(Penaeus vannamei)幼虾(体长为4.2~5.0cm),从组织细胞学水平对分泌蜕皮激素的Y-器官结构进行初步研究,观察KK-42处理后幼虾Y-器官的形态结构变化。结果表明,伴随动物的生长,Y-器官腺细胞条索变粗,腺细胞核增大。KK-42处理后腺细胞核的结构、大小未见显著变化,腺细胞索的宽度明显高于同一时期对照组。     

进境斑节对虾检疫和防治

本文系统地报道了进境斑节对虾亲虾及育苗期疫病的检疫方法、结果和防治措施。检出了多种致病性弧菌、微孢子虫和斑节对虾杆状病毒。细菌分离药敏试验和敏感药物防治是当前对付虾细菌病的先进方法。针对当前虾病,提出了加强检疫、建立育苗场用药指南、创造良好的育苗环境、防止虾病发生、培育壮苗的对策。     

日本对虾内陆养殖的实验室模拟研究

我们将日本对虾进行实验室桶养作模拟内陆养殖试验。发现日本对虾可以顺利地养殖到体长５－７厘变,然后短人存活率急剧降低,这一现象与我们海边虾塘养殖所得到的结果极为一致,我们称之为“５－７”现象。     

饲料中维生素A水平对凡纳滨对虾生长、饲料利用、体组成成分及非特异性免疫反应的影响

本文旨在研究5种等氮等能饲料中,维生素A添加水平对凡纳滨对虾(Penaeus vannameiBoone)生长、饲料利用、体组成成分及非特异性免疫反应的影响。试验选用平均初始体重为(0.069±0.005)g的凡纳滨对虾450尾,随机分为5个处理组,每组90尾,每组设3个重复,每个重复30尾。试验采用单因素设计,以维生素A醋酸酯为维生素A源,用添加不同水平维生素A(添加量分别为0、18、365、4和72 mg/kg)的饲料喂养凡纳滨对虾,试验期11周。结果表明:在养殖前4周,饲料中添加维生素A显著促进凡纳滨对虾的生长(P<0.05),提高其对饲料利用效率(P<0.05),而对对虾的成活率不产生显著的影响(P>0.05);第11周的结果表明,饲料中添加维生素A并不能显著影响凡纳滨对虾的生长、饲料利用效率和成活率(P>0.05)。维生素A对对虾体脂肪、蛋白质含量的影响显著(P<0.05),对对虾体水分和灰分含量无显著影响(P>0.05)。维生素A对对虾血清中溶菌酶活力、酚氧化酶活力和血液红细胞数量的影响显著(P<0.05),饲料中未添加维生素A或过量添加(超过36 mg/kg)均导致血清中溶菌酶活力、酚氧化酶活力和血细胞总数显著下降(P<0.0 5),而对血清超氧化物歧化酶活力无显著影响(P>0.0 5)。用折线回归模型分析饲料维生素A添加量与对虾增重率的变化关系,表明在试验前4周,凡纳滨对虾饲料中维生素A的适宜添加量为2 2.5 0 mg/kg;1 1周试验期间,凡纳滨对虾饲料中维生素A适宜添加量为1 8 mg/kg;以溶菌酶、酚氧化酶活力为指标,凡纳滨对虾饲料中维生素A最适添加量为5 9.5 1 mg/kg。     

全封闭生态养殖斑节对虾Penaeus monodon Fabricius技术的研究

本文报道了深圳市宝安区水产科学研究所，于１９９８年和１９９９年连续二年，在珠江口低盐区，采用全封闭生态养殖斑节对是技术，并取得成功，这项新技术为预防虾病提供理论基础和实践经验，具有推广和应用价值。