Sperm cryopreservation of Indian major carp, Labeo rohita: cryodiluents, sperm: cryodiluent dilution ratio and cryoprotectan t concentration

Cryogenic preservation trials of spermatozoa of Labeo rohita were carried out. Twenty four cryodiluents (extender + cryoprotectant), with the combination of six extenders such as egg-yolk citrate, urea-egg-yolk, 0.9% NaCl, Kurokura-2, Ma and Mb and four cryoprotectants viz. DMSO, glycerol, methanol and ethanol, were used to screen out the suitable cryodiluents. Sperm was preserved in 0.25ml plastic straw in programmable freezer. Two step freezing method was followed. Sperm preserved with egg-yolk citrate and urea-egg-yolk containing 10% DMSO showed best post-thaw motility (80%) followed by 0.9% NaCl (60%) and Kurokura-2(30%) solutions. Sperm with the extenders M" and Mb clotted at the time of equilibration and also after few days of preservation. Egg-yolk citrate mixed with ethanol and methanol also showed good percentage of motility (80%) but egg-yolk citrate with glycerol showed less sperm motility (>60%). To determine suitable dilution ratio of milt and cryodiluent two best extender eggyolk citrate and urea-egg-yolk with four cryoprotectants such as DMSO, glycerol, methanol and ethanol at different ratio viz 1:2,1:4,1:7,1:10,1:15 and 1:20 were used. Highest post-thaw motility (>80%) was observed when milt was preserved with egg-yolk citrate containing 10% DMSO at 1:2, 1:4, 1:7 and 1:10 dilutions. Meanwhile using glycerol as cryoprotectants provided less post thaw motility at lower dilution ratio but with the increase of its dilution showed good sperm motility compared with other cryoprotectants. Finally, evaluation on the effect of cryoprotectant concentration on post-thaw sperm motility was conducted. Egg-yolk citrate and four cryoprotectant i.e. DMSO, glycerol, methanol and ethanol with six different concentrations namely 5%,7%, 10%, 15%, 20% and 30%.were evaluated. Among the cryoprotectants DMSO, methanol and ethanol showed highest post-thaw motility (about 80%) at 7% and 10% concentrations. Although glycerol was not suitable at low concentration but its 20% and 30% concentration levels provided best post-thaw motility. No post-thaw motility was obtained with DMSO at 30% concentration. The overall analysis on cryoprotectant concentration indicated that below 5% and above 20% cryoprotectant concentrations could not be suitable for effective cryopreservation of spermatozoa.

Studies on the suitability of producing value-added products from silver carp, Hypophthalmichthys molitrix

Silver carp, Hypophthalmichthys molitrix is contributing significantly to the total production of fish through aquaculture in Bangladesh. However, its low market price has become a serious concern to the fish farmers. The suitability of silver carp mince for the production of various value-added products (VAPs) - surimi, fish sausage, fish burger and fish stick was studied during April-September 2000 to ensure more appropriate and profitable utilization of silver carp. Surimi/frozen mince block was produced by washing the silver carp mince with 0.1% NaCl for 7-8 min (4-5 min agitation and 3-4 min settling). A two-step heating schedule for incubation at 50°C for 2 h and cooking at 95°C for 30 min gave high textured good quality consumer product. With the addition of cryoprotectants, surimi could be kept frozen for 5 months without loosing [sic] much of its textural and sensory qualities. Mince-mix and a batter with different ingredients and spices were formulated to produce fish burger using potato smash as the binding agent. Fish flake-mix and a batter with different ingredients and spices were formulated to prepare fish stick using both potato starch and potato smash as filler ingredients. Unwashed and washed frozen mince block or fresh flesh of silver carp was used to prepare fish sausage by heating at 100°C for 1 h after incubating at 50°C for 2 h. A spice-mix formulated with various local spices at the rate of 1.0-1.2% gave good texture and flavor to the sausage. A good-appeared sausage-pink color was developed by combining three food-grade colors of asthaxanthin. Products prepared with potato starch, potato smash and rice smash had an acceptable bacterial load in refrigeration (5°C) for up to 8 days and in room temperature (28°C) for up to 3 days. No coliform bacteria were found in the products prepared.