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.     

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.     

中草药对斑节对虾生长、饲料利用和肌肉营养成分的影响

研究了中草药对斑节对虾Penaeusmonodon(初始体重0.30g.尾-1)生长性能、饲料利用以及肌肉营养成分和氨基酸的影响。6种实验饲料(按顺序分别为1,2,3,4,5和6)中中草药的添加量分别为0,0.5,1.0,2.0,4.0和8.0g·kg-1饲料。除了饲料2之外,斑节对虾的成活率、饲料系数和蛋白质效率都显著(P<0.05)优于对照组(饲料1)。投喂饲料3的斑节对虾增重率与对照组相比,虽然统计学上不存在显著差异,但有较大幅度的提高。投喂添加中草药的饲料对斑节对虾肌肉的水分和蛋白质含量无显著影响,但脂肪含量明显下降。结果表明,斑节对虾饲料中添加适量的中草药能够促进生长、显著提高成活率和降低饲料系数,并改变斑节对虾肌肉中脂肪含量和氨基酸组成。     

Dietary folic acid requirement determined for grass shrimp, Penaeus monodon

A feeding trial was conducted to determine the dietary folic acid requirement of juvenile grass shrimp, Penaeus monodon. Purified basal diets were formulated using vitamin-free casein as the protein source. Graded levels (0, 1, 2, 5, 10, 15, 30, and 60 mg folic acid/kg diet) of folic acid were added to the basal diet, resulting in eight dietary treatments in the experiment. Each diet was fed to three replicate groups of shrimp initially averaging 0.79±0.01 g for 8 weeks. Shrimp fed diets supplemented with 2 mg folic acid/kg diet had greater (P<0.05) weight gain and hepatopancreatic folic acid concentrations than shrimp fed the unsupplemented control diet. Feed efficiency (FE) was highest in shrimp fed the diets supplemented with 2 mg folic acid/kg diet, followed by the groups fed 1 mg/kg, then the control group. Hepatosomatic index (HSI) was highest in shrimp fed the control diet, followed by 1 mg folic acid/kg and lowest in shrimp fed diets supplemented with 2 mg folic acid/kg. Weight gain percentage, hepatopancreatic folic acid concentration and hepatosomatic index of the shrimp analyzed by broken-line regression indicated that the adequate dietary folic acid concentration in growing P. monodon is 1.9–2.1 mg/kg diet.     

斑节对虾GLUT1基因cDNA的克隆与表达分析

该研究利用RACE技术克隆获得了斑节对虾(Penaeus monodon) GLUT1 (glucose transporter type 1)的cDNA全长序列;采用实时荧光定量的方法研究了PmGLUT1在斑节对虾幼体发育过程中、各个组织中及低盐胁迫下的差异表达情况。该基因cDNA开放阅读框(ORF)全长1 476 bp,可编码491个氨基酸。检测PmGLUT1基因从受精卵至仔虾期发育过程的表达情况,结果显示,PmGLUT1在幼体发育各期的表达量有所波动,但总体呈现上升趋势。组织表达分析发现,PmGLUT1在鳃组织中的表达量最高,肝胰腺次之,在卵巢中的表达量最低。急性低盐胁迫后,PmGLUT1在肝胰腺中的表达水平显著高于对照组(P<0.05),但在鳃中的表达水平显著低于对照组(P<0.05)。研究结果表明,Pm GLUT1可能在斑节对虾幼体发育及机体应对低盐胁迫过程中具有重要作用,这为进一步研究葡萄糖转运蛋白基因在斑节对虾幼体发育调控和耐低盐胁迫应答中的分子机制提供了理论依据。     

Gene Expression in Black Tiger Prawns Following Intramuscular Injection of β-gal Plasmid

Gene expression in black tiger prawns (Penaeus monodon) was studied following intra-muscular injection of CMVGal plasmid into the second abdominal segment. We used an in situ staining technique to detect -gal expression in one- and three-month-old injected prawns. We found that only one of the three-month-old prawns expressed the marker gene (2 days after injection), and the site of expression was confined to the sixth abdominal segment away from the injection site. We repeated the experiment on a new batch of three-month-old prawns, but using fluorometric determination technique. This time we found that -gal expression was detected (6/42) at the site of injection after 2, 7, and 14 days. In two other test samples, transgene expression was detected in the sixth abdominal segment only, further confirming the possibility of injected DNA dispersal. The results of the study also suggest that direct gene transfer is a feasible technique in black tiger prawns.     

Dopamine modulates the physiological response of the tiger shrimp Penaeus monodon

Levels of glucose, lactate, pO₂, pCO₂, HCO₃⁻, TCO₂, Na⁺, K⁺, Cl⁻, protein, and oxyhemocyanin in the hemolymph and its osmolality and pH were measured when tiger shrimp, Penaeus monodon (13.5 ± 1.5 g body weight), were individually injected with saline or dopamine at 10^− 8, 10^− 7, or 10^− 6 mol shrimp^− 1. Results showed that hemolymph glucose, lactate, pCO₂, HCO₃⁻, and TCO₂ values increased from 2 to 4 h; hemolymph osmolality, Na⁺, and total protein had increased at 2 h; and hemolymph K⁺ decreased from 2 to 8 h after the dopamine injection. All physiological parameters returned to the control values 4–16 h after receiving dopamine. The dopamine injection also significantly decreased the oxyhemocyanin/protein ratio of P. monodon which occurred at 2 h, resulting from an elevation of hemolymph protein and a slight decrease of oxyhemocyanin. These results suggest that stress-inducing dopamine caused a transient period of modulation of energy metabolism, osmoregulation, respiration, and the acid–base balance in P. monodon in adapting to this environmental stress.