Larval rearing of Macrobrachium rosenbergii (De Man) in artificial sea water

Larvae of Macrobrachium rosenbergii were successfully reared in artificial sea water prepared in fresh ground water. The water was circulated through a biological filter by means of air-lift pumps for a period of one week to remove the undissolved particles prior to use in the hatchery operation. The experiments were initiated during 1989 and the hatchery has been working on pilot scale since June, 1990. The larvae in all the experiments were fed with egg-custard, Mona and Artemia nauplii. The survival rate varied from 5 to 52% in the 12 experiments. These findings can add to the development of hatcheries in the inland areas which can further boost the popularization of giant freshwater prawn farming.

Observations on the mating behavior, spawn mass and larval development of Hydatina physis Linne, 1758 (Cephalaspidea: Hydatinidae) from Karachi

Reproduction of Hydatina physis was studied in a population from Karachi, Pakistan, including mating and egg laying behavior, spawn characteristics and development.Individuals first appear in the field in October and remain until March. The spawning occurs from mid-November till mid-February with a peak in December. During this period the individuals were also observed pairing. In captivity, mating lasts for 30 minutes, second mating occurs two days later. Oviposition occurs in a very interesting and unusual manner. The mother turns "up-side-down" with its food fully expanded and the shell completely hidden underneath, the expanded foot serves as protective cover to the eggs. Eggs are deposited in a complexly folded mass with a short stem and an adhesive disc. Capsules, arranged in a single layer, contain 4-6 eggs each of wich is 70 um in diameter. Development is planktotrophic and veligers hatch after 14 days at a temperature of 26-28 degrees Celsius.

Rearing of Clarias batrachus (Lin.) larvae with formulated diets

An experiment was carried out for a period of 20 days using 7-day old Clarias batrachus larvae of initial total length of 7.4 ± 0.49 mm and weight of 2.9 ± 0.83 mg. Three artificial diets were used for the study having three replication of each. Among these, diet-I was formulated using 20% fish meal (FM), 30% powdered milk and 30% boiled egg yolk (BEY), diet-II using 27% FM, 20% Baker’s yeast (BY), 30% BEY and 3% agar and diet-III using 20% FM, 20% BY and 45% whole egg. The larvae fed on diets-II and III showed significantly (P<0.05) better length and weight gain than those of the larvae fed on diet-I. The larvae fed on diet- III showed the best survival rate (70%). However, the condition factor of the larvae fed on diet I was significantly better than those of the larvae fed on other two diets. The results of the study showed that C. batrachus larvae could be successfully reared with diet containing 45% whole egg, 20% yeast and 20% fish meal.

Hatching and rearing of carp (Cyprinus carpio L.) in the Polonnaruwa nursery

Carp fingerlings have been raised at Polonnaruwa since 1957 (Ling, 1962), by a method essentially the same as that described by Hora and Pillay (1962). The present work was initiated to assess and increase the efficiency of the nursery. Two experiments were carried out. In the first, 3 females and 6 males were used. Thirty bundles of Hydrilla were tied to the 3 strings and 10 of them taken at random were used for egg counts. In the second experiment the same number of fish was used but 36 bundles of Hydrilla were tied to the 3 strings and 9 of these taken at random for egg counts. The results of these 2 experiments are given in Table l.

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.

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.

Cryopreservation of rhesus macaque (Macaca mulatta) spermatozoa and their functional assessment by in vitro fertilization

Although spermatozoa from several species of nonhuman primates have been cryopreserved, there has been no report of success with rhesus macaque spermatozoa as judged by functional assays. Two Tris-egg yolk freezing media. TEST and TTE. which have: been successfully used for cynomolgus macaque (Macaca fascicularis) spermatozoa, were compared for cryopreservation of spermatozoa From four rhesus macaques (Macaca mulatta). The postthaw motility (percentage and duration) of spermatozoa cryopreserved in TTE was much higher than that for spermatozoa cryopreserved in TEST. The function of sperm cryopreserved in TTE was evaluated by in vitro fertilization or oocytes collected from gonadotropin-stimulated prepubertal rhesus macaques. Of the inseminated oocytes. 82 +/- 13% were fertilized and 63 +/- 22 and 39 +/- 21% of the resulting zygotes developed into morulae and blastocysts. respectively. These results indicate that rhesus macaque spermatozoa can be effectively cryopreserved in TTE medium. This finding will facilitate the application of in vivo and in vitro assisted reproductive technologies in this species. (C) 2001 Academic Press.

Growth and reproductive performance of locally isolated brackish water rotifer (Brachionus plicatilis) feeding different micro algae

Population growth and reproductive capacity of brackishwater rotifer, Brachionus plicatilis, were evaluated, for a period of 8 days in a temperature controlled ( =25°C) microalgallaborarory, under three different algal feeding regimens. The algal species that were tested are: (i) Chlorella sp. (T1), Tetraselmis chui (T2), Nannochloropsis oculata (T 3). The feeding density of each algal species was maintained similar as of 4.5xW6 ceHs mi. The rotifer fed on T. chui showed the highest (p<0.05) population growth (131.5 ind./ml), compared to that fed on Chlorella sp (45.67 ind./ml) and N oculata (43.44 ind./ml). The abundance of egg bearing rotifers was also higher (35.77%) with T. chuithan with Chlorella sp (27.76%) and N oculara (24.60%). The results of the present study indicate that T. chui could be the most suitable algal food for the stock culture of locally isolated rotifer B. plicatilis.

Estimation of micro-flora associated with different stages of Macrobrachium rosenbergii (de Man)

The bacterial flora occurring in muscle, haemolymph, hepatopancreas and gill of brood, juveniles, water, eggs, larvae and rearing water were estimated by selective plate count technique for Entrobacteriaceae, Streptococaceae and Vibrionaceae members. The total viable bacterial count was estimated by total plate count technique on nutrient agar. The total viable counts of bacteria were lowest in water from 6.10x10² CFU/mL) and highest in egg (6.06x10super(8) CFU/g). In brood the total counts were varying from 1.62x10² CFU/g in muscle to 2.20x10super(5) CFU/g in gills. In juveniles, the total plate counts were varying from 2.8x10super(4) CFU/g in muscles to 3.67x10 super(8) CFU/g in hepatopancreas. Selective plate counts show that Enterobacteriaceae members dominate in egg and gills of brood and hepatopancreas of juveniles. Vibrios were found to be dominant in water and larvae of rearing tank. Haemolymph of brood was sterile and did not contain any bacteria while muscle of juvenile was having very low count of total viable bacteria.

Nutritional quality of some food fish

The proximate compositions and amino acid make-up of silver jew fish (Johnius argentatus), Indian halibut (Psettodes erumei), grey mullet (Mugil cephalus) and pearl spot (Etroplus suratensis) are reported. Calorific values of these fishes have been calculated from their proximate compositions and their amino acid make-up compared with the available data for beef and egg. From the study, pearl spot is adjudged to be the most nutritive among the fishes studied, followed by Indian halibut, grey mullet and silver jew fish.

Characterization and cloning of a novel phospholipase A(2) from the venom of Trimeresurus jerdonii snake

A phospholipase A(2) (PLA(2)) called jerdoxin, was isolated from Trimeresurus jerdonni snake venom and partially characterized. The protein was purified by three chromatographic steps. SDS-polyacrylamide gel electrophoresis in the presence or absence of dithiothreitol showed that it had a molecular mass of 15 kDa. Jerdoxin had an enzymatic activity of 39.4 mumol/min/mg towards egg yolk phosphatidyl choline (PC). It induced edema in the footpads of mice. In addition, jerdoxin exhibited indirect hemolytic activity. About 97% hemolysis was observed when 2 mug/ml enzyme was incubated for 90 min in the presence of PC and Ca2+. No detectable hemolysis was noticed when PC was not added. Ca2+ was necessary for jerdoxin to exert its hemolytic activity, since only 52% hemolysis was seen when Ca2+ was absent in the reaction mixture. Furthermore, jerdoxin inhibited ADP induced rabbit platelet aggregation and the inhibition was dose dependent with an IC50 of 1.0 muM. The complete amino acid sequence of jerdoxin deduced from cDNA sequence shared high homology with other snake venom PLA(2)s, especially the D49 PLA(2)s. Also, the residues concerned to Ca2+ binding were conserved. This is the first report of cDNA sequence of T jerdonii venom PLA(2). (C) 2002 Elsevier Science Ltd. All rights reserved.

A new observation on large scale disintegration of eggs of Indian major carps during development

While carp themselves may destroy carp eggs by sucking from outside the happa, destruction in the absence of carp also occurs. During observations of the effect of electrical field on developing carp eggs it was seen that the egg membrane ruptured prematurely under certain potential differences. In natural waters and mud potential differences occur in the presence of high concentrations of certain salts compared with the internal concentration. Temperature and pH fluctuations following strong sun or heavy rain accelerates the diffusion of ions along side water through the egg membrane. The egg cells expand rapidly and the embryos were rapidly dropped.

Cholesterol content in the eggs of some freshwater teleosts

The authors report the distribution of cholesterol in the ripe, unspawned eggs of 10 spp of teleost (carps, catfish and murrels) together with values for egg diam. The egg, with its relatively abundant yolk, is shown to be fairly rich in cholesterol, though values differ from sp to sp. The lowest value (700 mg/100 g) was recorded from Wallago attu, and the highest (1537 mg/100g) from Heteropneustes fossilis. In general the levels are lowest in carp eggs. No definite relationship could be established between lipid and cholesterol contents, or between egg diam and total cholesterol. The role of cholesterol in physiology of fish is discussed; it is noted that the carnivorous fish tended to have higher levels of cholesterol in their eggs.

Influence of sex, spawning, starvation and water temperature on fatty acid composition in Tilapia mossambica

The influence of sex, spawning, starvation and water temperature on the fatty acid composition of Tilapia mossambica has been studied. Tilapia egg lipid was found to have unusually high percentage of C sub(22:6) fatty acids (9.09%) compared to body and intestinal lipids. The C sub(16:1) acid was much less in the egg lipids (3.5%) whereas it was 11% in the body lipids. There was no significant difference in the fatty acid composition of body and intestinal lipids of male and female tilapia. Starvation caused the presence of high content of lower fatty acids (C sub(6), C sub(8), C sub(30), C sub(12) and C sub(33)) in the body lipids. Water temperature also influenced the fatty acid composition of Tilapia; the difference was more significant in body lipids than in intestinal lipids.

Studies on the toxicological effects of Asulox-40 and Emisan-6 to eggs and early life history stages of Sarotherodon mossambicus

Toxicological effects of Asulox-40 and Emisan-6 to eggs and early life history stages of Sarotherodon mossambicus were reported. 80% of egg hatching occurred in the controls, 1 p.p.m and 5 p.p.m concentrations of Asulox-40. 10 p.p.m. and 50 p.p.m. concentrations of the same toxicants had 70% and 60% hatchings while in Emisan-6 in the same concentrations the hatching were 70% and! 40%. In 100 p.p.m. concentration of both toxicants 20% incomplete hatching occurred. In Emisan-6 Lc 50 and Lc 100 values were recorded at 32 hand 96h respectively in 10 p.p.m. concentrations. In Asulox-40 the same values were recorded in 24h and 40h respectively at 50 p.p.m. concentration. The fish activity during the experimental period showed initial hyper activity. It was established that the Emisan-6 is more harmful to S. mossambicus than Asulox-40. The harmless concentrations of these chemicals were 1.2 p.p.m. for Asulox-40 and 0.6 p.p.m. for Emisan-6.