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.     

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。     

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。     

利用塑料大棚延长对虾交尾期的研究

为提高对虾交尾率，１９９３，１９９４年进行了塑料大棚延长对虾交尾期试验，通过大棚控制交尾水温，采取合理安排雌雄比，调节水质，投喂沙蚕，防治疾病等综合技术措施，２年共出棚交尾虾１２３５２尾，未交尾虾５９２２尾，雄虾５２５２尾，平均交尾率６７．５９％，雌雄虾成活率分别为８７．５４％，６３．００％，共获产值６２６１００元，纯利４４２６００元，平均每尾入棚雌虾净协效益２１．２３元，专家鉴定认为，本研究达国     

日本对虾的实验室养殖研究

我们曾将发病残存的日本对虾继续养殖作为亲虾，进行夏季高温育苗，所得到的虾苗在实验室里桶养，证明在人工饲养条件下它们是可成活和生长的，本实验还提示，若能解决水质净化问题，日本对虾甚至有可能移至内陆养殖。