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。     

斑节对虾循环水养殖系统构建与运行试验

为探索斑节对虾循环水养殖可行性及应用发展价值,该研究自主设计蛋白分离组合装置、内循环流化床等关键工艺环节水净化装备,构建了技术先进、结构紧凑的斑节对虾循环水养殖系统。针对其不同阶段生长特性及水环境需求,提出一种水质调控方法,科学投喂。运行试验120d,溶解氧浓度5.30～7.14mg/L,p H值7.23～8.44,氨氮浓度0.43～1.38 mg/L(稳定运行后),亚硝酸盐氮浓度0.15～0.56 mg/L(稳定运行后);斑节对虾在循环水养殖模式水生态环境下正常生长,先后经历快速生长期、稳定生长期及缓慢生长期,终末养殖密度3.02 kg/m2,取得高效养殖结果;终末饲料系数1.67,单茬每平方米利润34.78元,每平方米年利润69.56元(按1年2茬计),获得良好经济效益。该实践可为斑节对虾循环水养殖模式应用提供技术支持。     

Effects of dietary highly unsaturated fatty acids and astaxanthin on the fecundity and lipid content of pond-reared Penaeus monodon (Fabricius) broodstock

Five diets that contained fresh squid meat as the basic constituent and were supplemented with different amounts of highly unsaturated fatty acids (HUFA) and astaxanthin were fed to pond‐reared Penaeus monodon broodstock. Diet A was sole squid meat. Diets B and C were supplemented with astaxanthin 50 and 100 mg kg^?1 respectively. Diets D and E were supplemented with HUFA 5 and 10 g kg^?1 and astaxanthin 50 mg kg^?1 respectively. The result showed that the group fed diet E had the best reproductive performance in all experimental groups. It had a higher proportion of spawns (71.5%), spawning rate (0.047), a shorter latency period (7.7±0.3 d), higher absolute fecundity (× 10³) (361.6±5.5) and egg production/female (× 10³) (597.0±18.0) than all the other experimental groups. The fatty acid composition in broodstock diets strongly affected the tissue and fecundity of broodstock. Good correlations between the content of 20:4n‐6 in eggs and the fecundity (r²=0.6109) and egg production (r²=0.9876) of broodstock were found. On the other hand, 22:6n‐3 and DHA/EPA ratio was negatively correlated with the fecundity of broodstock (r²=0.5362, 0.8702 respectively). The result also showed that the balance between n‐3 and n‐6 fatty acid families, total polyunsaturated fatty acids and total saturated fatty acid and 20:5n‐3 (EPA) and 22:6n‐3 (DHA) may play vital roles in maturation and reproductive performance of P. monodon broodstock.     

参虾池塘高效混养技术

7-9月刺参夏眠期间正是日本对虾快速生长的季节,在对虾池中人工造礁,第1年4月放入规格为80～120头/ kg的海参苗1740 kg,每年7月放入体长0.7～1.0 cm的日本对虾虾苗1.2×105尾,将刺参与日本对虾混养.在养殖过程中,刺参不投喂,只用卤虫、人工饲料、杂色蛤和四角蛤蜊投喂日本对虾,严格控制养殖池内的杂鱼虾,保持水质新鲜,可提高日本对虾和刺参的成活率,利用两者饵料和空间互补性,可显著提高虾池的利用率和经济效益.     

热泵干燥南美白对虾品质特性与玻璃化转变关系研究

针对南美白对虾热泵干燥过程中常出现物理、化学反应导致虾肉品质下降,而其机理又尚未明确的问题,采用静态称量法研究了南美白对虾肉的解吸等温线,利用差示扫描量热仪测定了虾肉的玻璃化转变温度(Tg)。分别采用GAB模型与Gordon-Taylor方程拟合水分吸附与Tg数据。虾肉在热泵干燥温度35℃、风速1.5 m/s下进行干燥,每隔1.0 h取样,分析虾肉品质特性(色泽、收缩程度、硬度与韧性和虾青素质量比)。基于玻璃化转变理论,阐明虾肉热泵干燥品质变化机制。结果表明,虾肉热泵干燥过程中,基质处于橡胶态,体系流动性增加,导致干制品品质下降。     

白斑综合征病毒与凡纳滨对虾肠道菌群区系之间关系的初步研究

比较了同一养殖池中的感染白斑综合征病毒(WSSV)和未感染WSSV的凡纳滨对虾的肠道菌群,旨在探讨对虾肠道菌群与机体健康状态之间的关系。感染WSSV对虾肠道的细菌总数为1.06×106CFU/尾,显著高于未感染WSSV的对虾(1.78×105CFU/尾;P<0.05),且其肠道中的细菌分别属于弧菌属、气单胞菌属、海水球菌属、盐水球菌属和乳酸杆菌属;染WSSV的对虾(P<0.05),气单胞菌的比例显著的低于感染WSSV的对虾(P<0.05);盐球菌所占的比例两者之间差异不显著。结果显示,两种对虾在肠道菌群组成和细菌组成和细菌数量上存在显著的差异,初步表明对虾肠道菌群区系和机体的健康状态密切相关。