Maternal effects on fecundity and egg quality of the Patagonian stock of Argentine Hake (Merluccius hubbsi)

The influences of age, size, and condition of spawning females on fecundity and oocyte quality were analyzed for the Patagonian stock of Argentine Hake (Merluccius hubbsi). Samples of mature females were collected in the spawning area as part of 2 research surveys conducted in January 2010 and 2011, during the peak of the reproductive season. Batch fecundity (BF) ranged between 40,500 (29 cm total length [TL]) and 2,550,000 (95 cm TL) hydrated oocytes, and was positively correlated with TL, gutted weight, age, hepatosomatic index (HSI), and the relative condition factor (Kn). Relative fecundity ranged between 85 and 1040 hydrated oocytes g–1 and showed significant positive relationships with gutted weight, HSI, and Kn; however, coefficients of determination were low for all regressions. Dry weights of samples of 100 hydrated oocytes ranged between 1.8 and 3.95 mg and were positively correlated with all variables analyzed, including batch and relative fecundity. Multiple regression models created with data of the morphophysiological characteristics of females supported maternal influences on fecundity and egg weights. Within the studied size range (29–95 cm TL), larger individuals had better somatic and egg condition, mainly revealed by higher HSI and hydrated oocytes with larger oil droplets (275.71μm [standard error 1.49]). These results were associated with the higher feeding activity of larger females during the spawning season in comparison with the feeding activity of young individuals (<5 years old); the better nutritional state of larger females, assumed to result from more feeding, was conducive to greater production of high-quality eggs.

Cryopreservation of sperm of common carp, Cyprinus carpio and silver barb, Barbonymus gonionotus for genetically improved seed production

Experiments were conducted to develop and standardize the protocols for cryopreservation of sperm of common carp, Cyprinus carpio and also for using the cryopreserved sperm for fertilization of eggs. Nine extender solutions as Alsever's solution, kurokura-1, kurokura-2, urea egg-yolk, egg-yolk citrate, 0.6% glucose, 0.9% NaCl, Ma and Mb, and five cryoprotectants namely ethanol, methanol, dimethylsulfoxide (DMSO), dimethylamine (DMA) and glycerol were tested. The cryoprotectants were mixed at 10% concentration of the extenders (v/v) to make the cryodiluents. Milt and cryodiluents were mixed at a ratio of 1:9 for Alsever's solution, kurokura-1, kurokura-2, 0.6% glucose and 0.9% NaCl, 1:4 for urea egg-yolk, egg-yolk citrate, Ma and Mb. Among the cryodiluents Alsever's solution mixed with either ethanol or methanol was found to be suitable and it produced more than 90% and 80% spermatozoan motility at equilibrium and post-thaw periods, respectively. Kurokura-1 and kurokura-2 when mixed with the same cryoprotectants showed good spermatozoan motility at equilibrium period (80-90%) but the motility was reduced (30-55%) at post-thaw state. Other extenders did not produce acceptable sperm-motility and in some cases the frozen milt became clotted. Different dilution ratios (1:1, 1:2, 1:4, 1:5, 1:7, 1:9, 1:12, 1:15, 1:20) were formulated for obtaining a suitable milt dilution, the dilution ratio of 1: 9 (milt : cryodiluent) demonstrated the highest post-thaw spermatozoan motility (80%) in Alserver's solution. The optimum concentration of cryoprotectants in the cryodiluents was determined, 10% concentration level was found to be effective to produce the highest number of spermatozoan motility in comparison to the other concentrations (5%, 15%, 20% 30%). Sperm preserved with the cryodiluent Alsever's solution along with either methanol or ethanol was found to be effective to fertilize eggs and produce hatchlings. The hatching rates ranged between 1.48% and 14.76%, compare to control. The fish produced through use of cryopreserved sperm and normal sperm were found to grow well and no significant (P<0.05) growth difference was observed between them. In case of silver barb, Barbonymus gonionotus, sperm tested against six extenders such as egg-yolk citrate, urea-egg-yolk, kurokura-1, kurokura-2, 0.9% NaCl and modified fish ringer (MFR) solution. Cryoprotectants used were the same as those of C. carpio. Milt was diluted with the cryodiluent at a ratio of 1:4 for egg-yolk citrate and urea-egg-yolk, 1:5 for kurokura-1 and 1:9 for 0.9% NaCl, MFR and kurokura-2. The cryoprotectant concentration was maintained at 10% of the extender (v/v) in all the cases. Among the extenders, egg-yolk citrate and urea-egg-yolk mixed with 10% DMSO, methanol and ethanol produced 50% post-thaw spermatozoan motility, whereas DMA and glycerol provided only 10% motility. Trials on milt dilution ratio and cryoprotectant concentration are being conducted. Fertilization trials are also underway.