Re-evaluation of total sulphur amino acid requirement and determination of replacement value of cystine for methionine in semi-purified diets of juvenile Nile tilapia, Oreochromis niloticus

Two feeding experiments were conducted to re-evaluate the total sulphur amino acid (TSAA; methionine and cystine) requirement and determine the replacement value of cystine for methionine of juvenile Nile tilapia ( Oreochromis niloticus ). Semi-purified diets used in both experiments contained 3510 kcal gross energy and 280 g of protein per kilogram diet from casein, gelatin and crystalline amino acids. The basal diet of the first experiment contained 3.1 g methionine and 0.4 g cystine per kilogram. l -methionine was added to the seven remaining diets at 1.0 g kg⁻¹ increment to produce methionine levels ranging from 3.1 to 10.1 g kg⁻¹ diet. Each diet was fed to four replicate groups of juvenile Nile tilapia (1.28 g mean weight) in a recirculation system for 8 weeks. Broken-line regression analysis of weight gain data indicated that the TSAA requirement of juvenile Nile tilapia was 8.5 g kg⁻¹ of the diet or 30.4 g kg⁻¹ of dietary protein. In the second experiment, TSAA level was set at 95% of the requirement value determined in the first experiment. Seven diets were made with different ratios of l -methionine and l -cystine (20 : 80, 30 : 70, 40 : 60, 50 : 50, 60 : 40, 70 : 30 and 80 : 20, based on an equimolar sulphur basis). Each diet was also fed to four replicate groups of juvenile Nile tilapia (4.14 g mean weight) in a recirculation system for 8 weeks. Regression analysis of weight gain data using broken-line model indicated that cystine (on a molar sulphur basis) could replace up to 49% of methionine requirement in semi-purified diets for juvenile Nile tilapia.     

Antibody response of two populations of common carp, Cyprinus carpio L., exposed to koi herpesvirus

Common carp, Cyprinus carpio L., exposed to koi herpesvirus (KHV) may become persistently infected and populations containing such virus-infected individuals may transmit the virus to other fish when co-habited. Detection of virus-infected fish in a population is thus critical to surveillance and control programmes for KHV. A study was therefore designed to detect anti-KHV serum antibodies, with an enzyme-linked immunosorbent assay, in common carp following experimental exposures to KHV under varying environmental conditions. The study determined that a proportion of fish within a population experimentally exposed to KHV (at least 10–25%) develop high antibody titres (1/1600 or greater) to the virus, and this immunological response was detectable for several months (observed at the termination of the experiments at 65, 46 and 27 weeks post-exposure). Furthermore, this response was detected in one population of fish that did not succumb to a high level of mortality when maintained at water temperatures that were non-permissive for KHV. Elevating the water temperatures to permissive conditions for KHV resulted in recurrence of disease despite the presence of anti-virus antibodies, suggesting that serum antibodies alone are not protective under the conditions of our trials.     

Evaluation of Streptocephalus dichotomus nauplii as a larval diet for freshwater prawn Macrobrachium rosenbergii

This study was performed to assess the nutritional value of Streptocephalus dichotomus nauplii and compared with standard larval diet and parthenogenetic strain of Artemia nauplii. The effectiveness of live feed was determined by feeding freshwater prawn Macrobrachium rosenbergii postlarvae. Results on the growth rate, weight gain, survival, fatty acid and amino acid composition show a significant variation between the dietary treatments. Mean larval growth was significantly different ( P  < 0.001) between control diet (13.5 ± 0.5 mm) and live feeds ( Streptocephalus nauplii; 18.4 ± 0.5 mm and Artemia nauplii 18.7 ± 0.2 mm). The weight of larvae-fed Streptocephalus (41.1 ± 1.44 mg) and Artemia nauplii (41.7 ± 0.2 mg) was not significantly different; however, treatment with live feeds was significantly higher than the control (16.3 ± 0.5 mg) ( P  < 0.001). Proximate composition on the fairy shrimp reveals that they are rich in protein, lipid, essential amino acids (EAA) and essential fatty acids (EFA). The polyenoic unsaturated fatty acid (18 : 2 n-6 and 18 : 3 n-3) and highly unsaturated fatty acid (20 : 4 n-3 and 20 : 5 n-3) show a dramatic increase in larval tissue relative to its proportional composition in the live diets. Amino acid composition in the live feeds, Streptocephalus and Artemia nauplii, perfectly reflects on the amino acid proportions in the larvae-fed diets which confirm its utilization.     

A dietary energy level of 14.6 MJ kg−1 and protein-to-energy ratio of 20.2 g MJ−1 results in best growth performance and nutrient accretion in silver barb Puntius gonionotus fingerlings

Five iso-nitrogenous (300 g protein kg⁻¹ diet) and iso-lipidic (80 g kg⁻¹ diet) semi-purified experimental diets with variable energy levels of 10.5 (D-1), 12.5 (D-2), 14.6 (D-3), 16.7 (D-4) and 18.8 (D-5) MJ kg⁻¹ diets were fed to Puntius gonionotus fingerlings (average weight 1.79 ± 0.02 g) in triplicate groups (15 healthy fishes per replicate) for a period of 90 days to assess the optimum dietary energy level and protein-to-energy ratio (P/E). Fifteen flow-through cement tanks of 100 L capacity with a flow rate of 0.5 L min⁻¹ were used for rearing the fish. Maximum specific growth rate, protein efficiency ratio, protein productive value, RNA : DNA ratio, whole body protein content, digestive enzyme activity and minimum feed conversion ratio was found in fish-fed diet D-3 with 14.6 MJ kg⁻¹ energy level. There were no improvements in all these parameters with the further rise in dietary energy level. Hence, it may be concluded that the optimum dietary gross energy level for maximum growth and nutrient utilization of silver barb is 14.6 MJ kg⁻¹ diet with a resultant P/E ratio of 20.2 g protein MJ⁻¹ diet, when the dietary protein and lipid are maintained at optimum requirement levels of 300 and 80 g kg⁻¹ diet, respectively, for this species.     

The effectiveness of a commercial microbial product in poorly prepared tiger shrimp, Penaeus monodon (Fabricius), ponds

The efficacy of a commercial microbial product was tested in commercial tiger shrimp, Penaeus monodon (Fabricius), ponds for one culture period in Kuala Selangor, Malaysia. Four ponds with replicates for treatment and control were used. The pond bottom was dried but the organic sludge was not removed as normally practised in pond preparation. The ponds were stocked with 15 post‐larvae at the rate of 31.m^?2. Physical, chemical and biological parameters of the pond were analysed every 2 weeks during the culture period. Water quality parameters remained within the optimum range for shrimp culture except for ammonia‐nitrogen being significantly higher in control ponds and silica in treated ponds. Benthic organisms were not found in any of the ponds. The average counts of different bacteria were not significantly higher in treated ponds than control. Because of poor health, the shrimp were harvested earlier (72 days) than the usual 120 days. An average of 875.60 ± 67.00 kg shrimp ha^?1 was obtained in treated ponds with a feed conversion ratio (FCR) of 1.57 ± 0.10 and survival rate of 42.35 ± 5.37% compared with 719.50 ± 130.94 kg shrimp ha^?1, 2.99 ± 0.70 and 21.25 ± 3.26%, respectively, in control ponds. Neither the microbial product nor the frequent water exchange was effective in overcoming the problems caused by the poor pond bottom.     

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.     

Comparison of larval performance between Thai and Vietnamese freshwater giant prawn, Macrobrachium rosenbergii (de Man): a preliminary study

This study compared the performance of larvae of giant freshwater prawn (Macrobrachium rosenbergii) from Thailand and Vietnam. Thai broods were collected from a farm and Vietnamese broods were collected from a river. Larvae from these two sources were raised in identical environments, and their rates of development and survival as well as their ability to withstand environmental stresses were assessed. The first postlarval stage was observed in both strains on day 22. All larvae (100%) from Thai broods completed metamorphosis by day 36, but only 89% of the Vietnam strain completed metamorphosis by day 45. The stages of development were more uniform in larvae from the Thai stock than the Vietnamese stock. Survival differences were observed within the first week (92% in the Thai group vs. 55% in the Vietnamese group). These differences were most pronounced beginning in week 5 (52% and 16% respectively) and week 7 (33% and 2% respectively). No clear difference was found in brood size and fecundity between the two groups. No difference in survival was observed between the two groups of larvae challenged with salinity, oxygen or formalin. Differential development rates of larvae in the same batch would naturally create conditions favourable for cannibalism and impaired ability to capture food. The earlier and more uniform rate of development, and subsequent higher rate of survival of Thai stock compared with the Vietnamese wild stock is suggested to be due to inadvertent selection in the domesticated stock.     

Using a combination of SAR element and enhancer to increase the expression level of hybrid genes in transgenic animals

A regulatory element consisting of a MAR element sequence of Drosophila heat shock genes and enhancer from a long terminal repeat of the mouse mammary tumor virus is created for studying the possibility of increasing the expression level of hybrid genes in transgenic animals. Two lines of transgenic mice containing the described regulatory element and hybrid gene intended for expressing human beta interferon in milk of transgenic animals are obtained by the method of coinjection into the pronuclei of murine zygotes. It is shown that the use of the given regulatory element doesn’t lead to an increase of activity of human beta interferon in milk compared with transgenic rabbits containing only the hybrid gene for expressing beta interferon. Furthermore, a decrease of tissue-specific expression of beta interferon is observed with the use of the regulatory element.