Cryopreservation of Persian sturgeon (Acipenser persicus) sperm: effects of cryoprotectants,antioxidant, membrane stabilizer,equilibration time and dilution ratio on sperm motility and fertility

Three experiments were performed to develop protocols for cryopreservation of Persian sturgeon Acipenser persicus, sperm. In the first experiment, sperm from six males was individually split in three subsamples and cryopreserved using Modified Tsvetkova's extender (mT) supplemented with dimethyl sulfoxide (DMSO), methanol (MeOH), glycerol (Gly) and ethylene glycol (EG) at concentration of 5%, 10%, 15% and 20%. In the second set of experiments, the effects of six equilibration times (0, 5, 10, 20, 40 and 60 min) and dilution ratios (volume sperm: volume extender 1:0.5, 1:1, 1:2, 1:3, 1:5 and 1:10) and the additive advantage of bovine serum albumin (BSA; 0, 2.5, 5 and 10 mg mL^?1) and ascorbic acid (0, 2.5, 5 and 10 U mL^?1), on the post‐thaw survival of sperm (triplicate set of six fish) were evaluated. Then, sperm was diluted in 1:1 mT extender with 10 mg mL^?1 BSA with selected cryoprotectants (15% MeOH and 10% DMSO) for 5 min. After a month of storage in liquid nitrogen, post‐thawed sperm motility; fertilization and hatching rate and viability of derived larvae were measured (Exp.3). Evaluation of cryoprotectants efficiency showed that MeOH 15% and DMSO 10% were suitable for cryopreservation of Persian sturgeon sperm. Gly and EG resulted in very low post‐thaw motility rates even at lowest concentration. No significant difference was observed among the four different equilibration times (0, 5, 10, 20 min) (P > 0.05) although higher equilibration times than 20 min resulted low post‐thaw motility (P < 0.05). The motility of frozen–thawed sperm did not significantly change when dilution ratio was increased from 1:0.5 to 1:3 (P > 0.05). However, higher dilution ratios (1:5 and 1:10) reduced the percentage of motile sperm. Supplementation of the cryoprotectant solution with 10 mg mL^?1 BSA significantly improved post‐thaw motility (P < 0.05), but ascorbic acid did not improve post‐thaw motility (P > 0.05). The results of experiment 3 showed that the highest fertilization (30.2 ± 5.75) and hatching rates (28.2 ± 5.25) were observed when samples were frozen with 15% MeOH (P > 0.05). Our study indicates that the use of mT extender consisting of 10 mg mL^?1 BSA in 15% MeOH diluted with sperm at 1:1 ratio for 5 min can be recommended cryopreservation method for Persian sturgeon sperm.     

疣吻沙蚕胚胎发育观察及盐度对其孵化的影响

采用显微镜观察了疣吻沙蚕(Tylorrhynchus heterochaeta)胚胎发育不同时期特征,并依据疣吻沙蚕人工孵化经验设置6个盐度梯度组(A:0,B:5,C:10,D:13,E:15,F:20),每组3个平行,研究了盐度对疣吻沙蚕受精率和孵化率的影响。结果表明,疣吻沙蚕胚胎发育可分为受精卵、卵裂期、囊胚期、原肠期、前担轮幼虫期、后担轮幼虫期、膜内三刚节疣足幼虫期和三刚节疣足幼虫期8个时期;盐度对疣吻沙蚕孵化影响显著(P0.05),C组受精率和孵化率最高,与其他组存在显著差异(P0.05)。适宜受精的盐度范围为10~13,最适孵化的盐度为10。     

Effects of Salinity,Commercial Salts,and Water Type on Cultivation of the Cryptophyte Microalgae Rhodomonas salina and the Calanoid Copepod Acartia tonsa

Marine aquaculture facilities positioned far from the sea need access to seawater (SW); hence, commercial salts are often the chosen solution. In marine hatcheries, most fish larvae require live feed (zooplankton) that are in turn fed with microalgae. The objective of this research was to investigate the applicability of commercial salts and clarify the potential effects on the cultivation of the microalga Rhodomonas salina and the copepod Acartia tonsa. Three commercial salts were tested, Red Sea Salt (RS), Red Sea – Coral Pro Salt (CP), and Blue Treasure Salt. R. salina was cultured at salinities of 10, 20, and 30 psu resulting in equal growth rates at salinities 20 and 30 in SW and RS mixed with deionized (DI) water. The optimum salinity for R. salina was 29 psu. For A. tonsa eggs, we observed highest hatching success in 30 psu with CP or RS mixed with DI water. The egg hatching success was not affected by salinities 15–40 and optimal hatching was obtained at 27 psu. Results confirm it was possible to use commercial salts for rearing of both R. salina and A. tonsa, widening the application of these species for aquaculture facilities without access to SW.     

Analysis of sinking death using video images of the swimming performance of Pacific Bluefin tuna (Thunnus orientalis) larvae

In this study, we aimed to clarify the mechanism of sinking death during the larval stage of Pacific Bluefin tuna Thunnus orientalis by investigating the effects of swimming performance on sinking death, using a behavioral approach. Swimming performance was examined 3–9 days after hatching (DAH) under day and night light conditions in cuboid experimental tanks. Swimming behavior variables such as swimming speed and swimming angle were measured under both light conditions. Larvae in the daytime experiment and larvae with inflated swim bladders at night were distributed on the surface layer of the water column. In contrast, larvae with uninflated swim bladders at night were frequently observed swimming vertically or sinking to the bottom of the tank. Larvae with inflated swim bladders at night were always distributed beneath the surface until the next morning (survival rates were 100 %). However, larvae with uninflated swim bladders at night swam upward repeatedly and later sank to the bottom of the tank (survival rates were 60 % and 38 % at 5 and 9 DAH, respectively). Larvae with uninflated swim bladders were not always able to maintain their swimming depth by swimming until the next morning. Additionally, their swimming speed and vertical swimming frequency (ratio) depended on the illumination and swim bladder conditions. Our findings show that larvae with uninflated swim bladders at night were associated with a higher risk of sinking death. The swimming energy capacity of Pacific Bluefin tuna larvae, which indicates the total amount of the energy that enables individuals to swim throughout the night without feeding, was found to be linked to sinking death.     

Post‐hatching development of the ornamental ‘Red Cherry Shrimp’ Neocaridina davidi (Bouvier, 1904) (Crustacea,Caridea, Atyidae) under laboratorial conditions

The ‘Red Cherry shrimp’, Neocaridina davidi is a small freshwater caridean shrimp living, originally, in various kinds of inland water bodies around Asian countries. This shrimp has reached several countries for ornamental use; however, basic information on the biology of the species is still scarce in the literature. Its early post embryonic development morphology has not yet been described. This paper focused on the production and the development of early post‐hatching stages of N. davidi, its male secondary sexual features, observation of the gonads and the presence of females with embryos. The larval development of N. davidi was almost suppressed as noted by the presence of relatively large‐sized eggs, first stage hatching as a decapodid; and the tail fans were present only from the 2nd post‐hatching stage. A biological important consequence of the presence of this Red Cherry shrimp species in the Neotropics is its potential release into nature, which could cause its rapid dispersion affecting populations of other indigenous caridean freshwater shrimps.     

Pectin purified from pomelo (Citrus maxima) peel as a natural hatching agent for fish embryos

Pectin is a biodegradable polysaccharide, and it has been recently applied as a gene delivery, drug delivery, wound healing and tissue engineering agent. In this study, pectin was extracted from pomelo (Citrus maxima) peel and characterized. The extraction recovery of pectin form pomelo peel was 14.5%, and it had 72.56% degree of esterification, 1,245.56 equivalent weight, 7.82% methoxyl and 68.27% anhydrouronic acid contents. Use of pomelo pectin as a hatching enhancing agent for fish embryos and its effect on hatching enzyme 1 (ZHE1) was investigated. The pectin‐exposed zebrafish embryos (100 µg/ml) showed significantly (p < .05) higher hatching rate (96.6%) compared with untreated (control) embryos (66.6%) at 60 hpf. The mRNA expression of ZHE1 was also significantly (p < .05) elevated up to 55.6‐fold in pectin‐exposed embryos at 24 hpf. In situ hybridization results revealed remarkably strong expression of ZHE1 in pectin‐exposed embryos compared with the control group. In addition, considerably larger size of the hatching gland was observed in pectin‐exposed larvae than that of the unexposed larvae group. These results clearly indicate that pectin isolated from pomelo peel has an ability to enhance the hatching process of zebrafish embryos via upregulation of ZHE1.     

中华沙鳅人工繁殖技术初探

对从长江、金沙江和岷江水域中收集质量为25.0～100.0 g的中华沙鳅(Botia superciliaris Günther)进行人工驯养、亲鱼培育和人工繁殖试验.结果显示:野生中华沙鳅能在人工饲养条件下生长和繁殖.注射人工催产药物鲤鱼脑垂体(PC)和促黄体素释放激素类似物(LHRH-A2)可促使亲鱼产卵和排精,催...     

温度和盐度对吉富品系尼罗罗非鱼受精率和孵化率的联合影响

采用中心复合设计和响应曲面方法研究了温度(T,℃)和盐度(S)对吉富品系尼罗罗非鱼人工受精率(FR,％)和孵化率(HR,％)的综合影响.设定的温度范围为20～34,盐度范围为0～18.建立了温度、盐度与受精和孵化之间关系的定量模型,并通过分析明确了温度与盐度的最优组合.结果表明,吉富罗非鱼在温度与盐度较高或较低范围内受精率与孵化率都处于较低水平.温度和盐度的一次与二次效应对受精与孵化均有极显著的影响(P＜0.01).温度与盐度二者间协同效应不显著(P＞0.05).建立了吉富罗非鱼受精率与孵化率的多项式方程,复相关系数分别达到了0.957 8和0.960 1 (P＜0.01),可以用于预测吉富罗非鱼的受精率和孵化率.利用统计优化技术明确,在温度27.14℃,盐度8.95时,罗非鱼最佳的受精率为80.8％,孵化率为77.63％.本研究旨在考察温度与盐度同时变化对吉富罗非鱼受精率与孵化率的影响,探讨最佳温度－盐度组合以增加吉富罗非鱼苗种产量.     

真蛸人工繁殖及受精卵孵化技术研究初报

真蛸为福建省近年来新兴的海水养殖优良种类，具有较高的营养价值、商品价值和产业化养殖前景。本文报道了福建省水产研究所2008年-2011年真蛸人工繁殖及受精卵孵化结果：127头雌亲蛸经强化培育，共有77头产卵，获卵194×10^4粒，孵出143．4×10^4头浮游幼体。雌亲蛸获产率46．2％～86．7％，卵子受精率75．6％-91．8％，受精卵孵化率77．9％～95．0％。在20．0～22．5℃水温条件下，真蛸受精卵经21～36d开始孵出，孵出后4～6d为孵出高峰，每窝卵开始孵出至孵化结束维持10—16d。