Carbohydrate level in the diet of silver barb, Puntius gonionotus (Bleeker) fingerlings: effect on growth, nutrient utilization and whole body composition

Five iso-nitrogenous (300 g crude protein kg⁻¹ diet) semi-purified diets with graded levels of carbohydrate at 220 (D-1), 260 (D-2), 300 (D-3), 340 (D-4) and 380 (D-5) g kg⁻¹ diet were fed ad libitum to Puntius gonionotus fingerlings (average weight 0.59±0.01 g) in triplicate groups (20 fish replicate⁻¹) for a period of 90 days to determine the effect of the dietary carbohydrate level on the growth, nutrient utilization, digestibility, gut enzyme activity and whole-body composition of fish. 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. The maximum weight gain, specific growth rate, protein efficiency ratio, RNA:DNA ratio, whole-body protein content, protease activity, protein and energy digestibility and minimum feed conversion ratio (FCR) were found in the D-2 group fed with 260 g carbohydrate kg⁻¹ diet. The highest protein and energy retention was also recorded in the same group. However, from the second-order polynomial regression analysis, the maximum growth and nutrient utilization of P. gonionotus fingerlings was 291.3–298.3 g carbohydrate kg⁻¹ diet at a dietary protein level of 300 g kg⁻¹ with a protein/energy (P/E) ratio of 20.58 −20.75 g protein MJ⁻¹.     

Protein-sparing effect of carbohydrate in silver barb, Puntius gonionotus fry

A 30‐day study was undertaken to examine the protein‐sparing effect of carbohydrate in diets for silver barb, Puntius gonionotus fry. Six semi‐purified experimental diets were formulated with two levels of protein (200 and 250 g kg⁻¹ diet) and three levels of carbohydrate (300, 340 and 380 g kg⁻¹ diet). In addition to the six experimental diets, a diet containing the protein and carbohydrate requirement levels of 300 and 260 g kg⁻¹ diet, respectively, as reported earlier for this species, was used as a reference diet. For each dietary treatment, 30 healthy fry of 20 days age (0.12 ± 0.01 g) were stocked in triplicate tanks using a flow‐through system. The fish were fed ad libitum four times a day to a level close to apparent satiation. Batch weighing of fish was done after 15 days of stocking to measure growth and general health status of the fish. The fish fed 250 g protein and 340 g carbohydrate kg⁻¹ diet with a protein to energy ratio of 17.86 g protein MJ⁻¹ performed equally well in terms of growth and nutrient utilization as the reference diet group. The study indicates that dietary protein can be reduced from 300 to 250 g kg⁻¹ diet by increasing carbohydrate from 260 to 340 g kg⁻¹ diet without sacrificing the growth of silver barb fry.     

Protein requirement of silver barb, Puntius gonionotus fingerlings

Five iso‐energetic (15.05 MJ kg^?1) semi‐purified diets with graded levels of crude protein, i.e. 200 (D‐1), 250 (D‐2), 300 (D‐3), 350 (D‐4) and 400 (D‐5) g kg^?1 diet were fed to Puntius gonionotus fingerlings (average weight 0.88 ± 0.03 g) in triplicate groups (15 healthy fish per replicate) for a period of 90 days to determine the optimum protein requirement of the fish. 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. Specific growth rate (SGR), food conversion (food gain) ratio (FCR), nutrient digestibility and retention, digestive enzyme activity, RNA : DNA ratio and tissue composition were used as response parameters with respect to dietary protein levels and feed intake. The mean weight gains of fish after 90 days were 10.84 ± 0.27, 11.07 ± 0.12, 14.09 ± 0.20, 11.27 ± 0.12 and 10.91 ± 0.25 g for D‐1, D‐2, D‐3, D‐4 and D‐5, respectively. Maximum SGR (3.13 ± 0.02% per day), RNA : DNA ratio (10.09 ± 0.09), tissue protein content (160 ± 0.1 g kg^?1 wet weight), protease activity (25.27 ± 0.47 μg of leucine liberated mg tissue per protein h^?1 at 37 °C) and minimum FCR (1.60 ± 0.02) was found in D‐3 group fed with 300 g kg^?1 protein level. All these parameters were negatively affected with the further increase in protein level in the diet. Digestibility of protein, lipid and energy was not affected because of variation in dietary protein levels and nitrogen intake of fish. Maximum energy retention (27.68 ± 0.12%) was recorded at 300 g kg^?1 dietary crude protein fed group. However, using broken line regression analysis, the maximum growth was found to be at 317.7 g kg^?1 dietary protein. Hence, it may be concluded that the protein requirement of P. gonionotus fingerling is 317.7 g kg^?1 diet with a resultant P/E ratio of 21.1 g protein MJ^?1.     

Effect of three different oil cake-based diets on pond production performance of silver barb, Puntius gonionotus (Bleeker)

One of the three iso-nitrogenous (30% crude protein) and iso-energetic (15.0 MJ kg⁻¹ diet) oil cake-based diets (groundnut, D-1; soybean, D-2 and mustard, D-3) was fed to the fingerlings (6.61 ± 0.06 g) of silver barb ( Puntius gonionotus ) for 120 days in triplicate fertilized ponds (0.04 ha) with a stocking density of 9000 fish ha⁻¹. Fish were fed at 10% of their body weight for the first month, followed by 3% in the second and 2.5% in the third and fourth months of the culture period. The fish fed diets D-1, D-2 and D-3 grew from initial weights of 6.67, 6.67 and 6.50 g to 320.23, 298.23 and 305.13 g respectively. No significant variation ( P >0.05) was found in the growth and nutrient utilization of fish fed different dietary treatments. Net profits of Indian Rupees 67 385, 59 535 and 65 790 ha⁻¹ were obtained in fish fed D-1, D-2 and D-3, respectively, within 120 days of culture. The results of this study suggest the possibility of the use of mustard oil cake as a dietary protein source for pond culture of silver barb in addition to groundnut and soybean oil cakes without compromising the growth, nutrient utilization, production performance and economics of culture of this fish.     

Development of cryopreservation methods for silver barb (Barbodes gonionotus,Bleeker) spermatozoa by manipulation of cooling rates in dry shipper

The aim of this study was to develop a simple cryopreservation protocol for silver barb, Barbodes gonionotus, semen using a dry shipper. Freezing rates within the upper and lower chambers of dry shipper were recorded for 14 days post liquid nitrogen loading (dpl). To regulate freezing rates, straws (250 and 500 µl) wrapped with various insulators (polystyrene foam box, oxygen tube, silicone tube and electric wire) were frozen within the upper chamber. Straws containing semen diluted with Calcium‐free Hank's Balanced Salt Solution (Ca‐F HBSS) and 10% dimethyl sulphoxide were cryopreserved with or without insulators. Appropriate protocols were selected based on sperm quality during a 45‐day cryostorage. The upper chamber had potential as a freezing chamber within 9 dpl due to no significant (p > 0.05) change in freezing rates. High percentages of sperm motility and viability (p < 0.05) were observed when 250 µl straws with silicone tube (T4) frozen for 5 min, non‐insulated 500 µl straws (T9) and 500 µl straws with polystyrene foam box (T12) frozen for 1–5 min, having freezing rates of 43.1 ± 1.3, 71.3 ± 1.4 and 14.7 ± 0.4°C/min respectively. Dry shipper can be used as a freezing tool to cryopreserve silver barb semen.     

Optimal dietary lipid level of silver barb, Puntius gonionotus fingerlings in relation to growth, nutrient retention and digestibility, muscle nucleic acid content and digestive enzyme activity

Five iso-nitrogenous (300 g kg⁻¹ diet) purified diets with graded level of lipid at 40 (D-1), 60 (D-2), 80 (D-3), 100 (D-4) and 120 (D-5) g kg⁻¹ diet were fed to Puntius gonionotus fingerlings for 90 days to determine their dietary lipid requirement. Two hundred and twenty-five fingerlings (average weight 2.34 ± 0.03 g) were equally distributed in five treatments in triplicate groups with 15 fish per replicate. 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. Specific growth rate (SGR), feed conversion ratio (FCR), nutrient digestibility, retention, digestive enzyme activity, RNA : DNA ratio and whole-body composition were considered as the response parameters with respect to dietary lipid levels. Maximum SGR and minimum FCR with highest RNA : DNA ratio, whole-body protein content and digestive enzyme activity was found in D-3 group fed with 80 g kg⁻¹ diet lipid. Nutrient digestibility was similar in all the groups irrespective of the dietary lipid level. Maximum protein and energy retention was recorded at 80 g kg⁻¹ dietary lipid fed group. However, from the second-order polynomial regression analysis, the maximum growth of P. gonionotus fingerlings was found at 96.9 g lipid kg⁻¹ diet.     

Apparent protein, lipid and energy digestibility coefficients of some commonly used feed ingredients in formulated pelleted diets for silver barb, Puntius gonionotus

To assess the apparent protein, lipid and energy digestibility coefficients of commonly used feed ingredients, viz. fish meal, groundnut oil cake, soybean meal, sunflower oil cake, sesame oil cake, mustard oil cake, rice bran, maize meal, black gram husk, green gram husk and wheat bran, a 50‐day trial was conducted for Puntius gonionotus fingerling (5.55 ± 0.32 g) in two successive phases using flow‐through circular fibre‐reinforced plastic tanks (60 L capacity). The photoperiod was 12 h light/12 h dark (12L : 12D). Ground water was used for rearing the fish. Chromic oxide (1%) was used as an external marker. The experimental fish were fed twice daily close to the levels of apparent satiation. The unconsumed feed and faeces were removed 1 h after each feeding. About 2 h after the removal of unconsumed feed and faeces, the freshly released faeces were collected twice daily by siphoning and were stored at ?18 °C for subsequent analysis. The apparent protein, lipid and energy digestibility coefficient values observed are in the range 81.88–95.60, 87.29–94.48 and 73.88–89.97%, respectively, for various test ingredients in P. gonionotus fingerlings, and the study suggests the usefulness of data for feed formulation.     

Evaluation of the potential source of bacterial contamination during cryopreservation process of silver barb (Barbodes gonionotus) sperm

Application of good sanitation practice in cryopreservation process is the key issue to improve the quality of cryopreserved fish sperm. This study implemented standard sanitation protocol with minimal contamination and evaluated the source of bacterial contamination associated with laboratory equipment and related materials across a series of cryopreservation process of silver barb (Barbodes gonionotus) semen. The use of 16s rRNA sequencing and traditional biochemical methods were performed for bacterial identification. Animal origin (anal fin, culture water and semen contaminated with faeces and urine) and non‐animal origin (liquid nitrogen from liquid nitrogen dewar, outer surface of straw, air circulation in cryopreservation laboratory and latex gloves used during cryopreservation procedure) were determined for bacterial contamination. Aeromonas punctata subsp. caviae was the most abundant species in anal fin, latex groves and semen contaminated with faeces and urine. Bacillus safensis and Bacillus sp. were found as frequently recovered species from liquid nitrogen dewar. Aeromonas hydrophila subsp. hydrophila and Pseudomonas fluorescens, fish pathogenic bacteria, were isolated from all animal origin samples. This was the first report indicating that standard sanitation and hygiene methodologies are recommended for sperm cryopreservation of B. gonionotus to prevent bacterial contamination.     

Compatibility of silver barb Puntius gonionotus (Bleeker) with Indian major carps in a grow-out polyculture

The compatibility and growth performance of silver barb Puntius gonionotus (Barbonymous gonionotus) with the three Indian major carps, i.e., Catla catla, Labeo rohita and Cirrhinus mrigala, were assessed in a 10‐month carp polyculture trial. Treatments T‐1, T‐2, T‐3 and T‐4 were stocked with three of the above four carp species, with an absence of silver barb, mrigal, rohu and catla, respectively, while all four species were stocked in treatment T‐5. The treatments were stocked at 6000 fingerlings ha⁻¹, with an equal species ratio maintained in each treatment. Incorporation of silver barb into the polyculture system neither affected the survival of any carp irrespective of species combination nor yielded significant changes in biomass production among treatments, except for the one without catla, where it was significantly low. The study revealed a higher extent of competition between silver barb and rohu, perceptible from the lower growth of one in the presence of the other. Although a certain level of competition of silver barb with mrigal was evident, competition with catla was not perceptible. Irrespective of species combination with silver barb as a component species, similar total biomass production in treatments revealed the feasibility of its incorporation into the Indian major carp‐based polyculture practice without affecting the total yield.     

Effect of extender composition and freezing rate on post-thaw motility and fertility of Arctic char, Salvelinus alpinus (L.), spermatozoa

The effects of extender composition and freezing rate on motility and fertility of frozen‐thawed Arctic char, Salvelinus alpinus, spermatozoa were investigated. Three freezing rates, two semen diluents and three cryoprotectants were tested. Semen frozen in 0.3 mol L^?1 glucose diluent with 10% methanol as a cryoprotectant or in a diluent described by Lahnsteiner with 10%N,N‐dimethylacetamide (DMA) resulted in the highest sperm motility. Fertility was the highest for semen frozen in a glucose–methanol extender but was not significantly different than that for semen frozen in Lahnsteiner's diluent with 10% DMA. Dimethyl sulphoxide (DMSO) at 10% was a relatively ineffective cryoprotectant with either semen diluent. Semen frozen at 6 cm above the surface of liquid nitrogen resulted in a higher post‐thaw sperm motility and fertility than semen frozen at 5 cm. The addition of 7% fresh egg yolk to glucose diluent containing methanol or DMSO did not improve the fertility of frozen‐thawed spermatozoa. However, the addition of 7% fresh egg yolk to glucose–DMA extender significantly improved the fertilization percentages of frozen‐thawed spermatozoa. In conclusion, dilution of semen 1:3 in 0.3 mol L^?1 glucose with 10% methanol and freezing 6 cm above the surface of liquid nitrogen (freezing rate of 40±8°C min^?1, mean±SD from ?5 to ?55°C) is a promising protocol for cryopreservation of Arctic char semen.     

Preservation of black sharkminnow,Labeo chrysophekadion (Bleeker, 1849) spermatozoa

The objective of this study was to investigate the effects of extenders and storage time on motility, viability and fertilization of preserved black sharkminnow, Labeo chrysophekadion spermatozoa. Sperm were diluted 1:3 in one of five extenders: modified Cortland solution (MC); Hanks' balanced salt solution (HBSS); 0.9% sodium chloride (NaCl); Kurokura solution (KU); and modified extender, and undiluted sperm samples were used as control and stored at 4°C for 5 days. Motility, viability and fertilization rates were evaluated every day. After a storage time of three days, the highest motility, viability and fertilization rates (61.27 ± 2.26%, 58.60 ± 2.29% and 40.58 ± 0.57, respectively) were achieved with sperm diluted with modified extender. Motility, viability and fertilization rates decreased significantly (P < 0.05) with increasing storage time in all treatments. In addition, this study found that motility, viability and fertilization had a positive significant correlation (P < 0.01). The results indicate that isotonic extender is suitable for the short‐term preservation of black sharkminnow spermatozoa.     

Sperm cryopreservation of silver barb (Barbodes gonionotus): cryoprotectants,cooling rate and storage time on sperm quality

Effectiveness and efficiency of frozen sperm on fertilization and hatching success of eggs from silver barb was examined in relation to cryoprotectants, freezing rate and storage period. Sperm was diluted in calcium‐free Hank's balanced salt solution, equilibrated with dimethylsulphoxide (DMSO), propylene glycol, sucrose or methanol at 5%, 10%, 15% or 20% final concentrations, and frozen in 250‐μL straws using a one‐step freezing procedure (1, 5 and 8°C min^?1 from 25 to ?40°C). Highest post‐thaw sperm motility was found from a treatment using 10% DMSO and 5°C min^?1 (82.2 ± 2.1%), similar to that of 10% DMSO and 8°C min^?1 (87.8 ± 3.2%). Post‐thaw motility of sperm frozen at 5 or 8°C min^?1 was significantly higher than 1°C min^?1. Relative sperm motility declined significantly after 10 months of cryostorage while viability did not change during a 12‐month cryostorage. Average fertilization rates of sperm after 1 and 4 months of storage were 64.5 ± 4.6% and 61.3 ± 3.4%, respectively, similar to those of fresh sperm (69.6–72.3%). Hatching rates of cryopreserved sperm (45.4–51.2%) were similar to those of fresh sperm (51.8–57.8%). This study developed suitable methods for cryopreservation of silver barb sperm that can be used to facilitate hatchery operation.     

Effect of different oil cake sources on growth, nutrient retention and digestibility, muscle nucleic acid content, gut enzyme activities and whole-body composition in silver barb, Puntius gonionotus fingerlings

Six iso‐nitrogenous (30% crude protein) and iso‐energetic (15 kJ g⁻¹) diets were prepared using different oil cake sources, viz. groundnut, soybean, sunflower, sesame, mustard and mixed oil cakes as major ingredients, and protein sources along with a minimum of 5% fish meal in each diet and were fed to silver barb Puntius gonionotus fingerlings (16.20±0.11 g) ad libitum four times a day close to an apparent satiation level for a period of 60 days to determine the effect of diets on growth, nutrient utilization, apparent digestibility coefficient (ADC) of the nutrients in the diets, gut enzyme activity, muscle nucleic acid content and whole‐body chemical composition of fish. Significantly higher (P<0.05) weight gain, specific growth rate, protein efficiency ratio, nutrient retention, ADC of nutrients in the diets, DNA:RNA ratio, protease and amylase activity with lower (P<0.05) feed:gain values were recorded in fish‐fed groundnut and soybean oil cake‐based diets than other diets tested. Among the dietary treatment groups, significantly higher (P<0.05) whole‐body protein, lipid and energy were also found in groundnut oil cake‐ and soybean oil cake‐based diets. The study suggests that the groundnut and soybean oil cake‐based diets, which led to significantly higher (P<0.05) growth and nutrient utilization than the other oil cake‐based diets in P. gonionotus fingerlings, may be used for pond culture of this species.     

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.     

Effect of cryoprotectants, extenders and freezing rates on the fertilization rate of frozen striped catfish, Pangasius hypophthalmus (Sauvage), sperm

The effects of four cryoprotectants (methanol, MeOH; dimethyl sulphoxide, DMSO; dimethyl acetamide, DMA; and ethylene glycol, EG), three extenders (calcium‐free Hanks' balanced salt solution, C‐F HBSS, Hanks' balanced salt solution, HBSS and sodium chloride, NaCl) and two different freezing procedures (one‐ and two‐step) on the cryopreservation of striped catfish (Pangasius hypophthalmus (Sauvage)) sperm were investigated. Sperm were frozen using a controlled‐rate freezer in 250 μL straws and stored for 2 weeks in a liquid nitrogen (LN₂) container. They were then airthawed at room temperature, and fertilization, motility and viability were assessed. The highest fertilization rate of 41% (81% of control) was achieved with the combination of 12% DMSO and 0.9% NaCl using a one‐step freezing procedure (10°C min^?1). Also, DMA resulted in a higher fertilization rate (30% or 51% of the control) than MeOH (18% or 38% of the control) or EG (8% or 12% of the control). In addition, the three extenders used did not affect fertilization rates after cryopreservation with each cryoprotectant. There were no significant differences among the three cryoprotectant concentrations and between the one‐ and two‐step freezing procedures. However, fertilization rates of cryopreserved sperm were significantly lower than the controls (P<0.05). The results of this study indicate that high fertilization rates of striped catfish eggs can be achieved using cryopreserved sperm when frozen at 10°C min^?1 in DMSO or DMA with either 0.9% NaCl or C‐F HBSS.     

Polyculture of silver barb, Puntius gonionotus (Bleeker), Nile tilapia, Oreochromis niloticus (L.), and common carp, Cyprinus carpio L., in Vietnamese ricefields: fish production parameters

The fish production parameters of five polyculture combinations, consisting of small and large silver barb, Puntius gonionotus (Bleeker), small and large Nile tilapia, Oreochromis niloticus (L.), and small common carp, Cyprinus carpio L., fingerlings in three replicates, were investigated in a rice-fish culture experiment (duration 149 days) conducted in the Mekong Delta, Vietnam. The survival rate was not significantly (P > 0.05) affected by the polyculture combination, but when grouped according to species, the mean survival of silver barb and tilapia was 64.3% and 63.7%, respectively, significantly higher than the mean common carp survival rate (33.4%). The growth of silver barb and tilapia was proportionally related to the stocking density, probably because of intraspecific competition and a synergistic interaction between silver barb and tilapia. The growth of common carp was not significantly different among the polyculture combinations. The highest net production (474.1 kg ha^?1) was obtained in the polyculture combination consisting of 80% small-sized silver barb fingerlings, but the fish was not marketable at that time. In concurrent rice-fish culture, it is recommended to raise large silver barb fingerlings. Small tilapia can be polycul-tured with silver barb, provided a stocking density lower than 1400 ha^?1. Common carp is considered less suitable because of a limited tolerance for the water quality conditions in the ricefield and the large size required by the market.     

Growth and survival of olive barb,Puntius sarana (Hamilton 1822) larvae produced with cryopreserved versus fresh sperm

Despite the success in fertilization and hatching of fish eggs with cryopreserved sperm, report on growth and survival of larvae produced from frozen‐thawed sperm is inadequate. The study evaluates the applicability of cryopreserved sperm for mass seed production by comparing the growth and survival of a popular food‐fish olive barb, Puntius sarana (Hamilton 1822) larvae produced from cryopreserved and fresh sperm. The eggs were artificially fertilized with cryopreserved and freshly collected sperm, and the growth and survival of produced larvae from both group recorded up to 12 weeks. The independent sample t‐test statistic showed the difference in lengths, t(718) = 0.241; P = 0.810 and weights, t(718) = 0.412; P = 0.680 were insignificant between two groups. There was also no significant difference, t(718) = ?0.758, P = 0.448 in survival of larvae produced from cryopreserved and freshly collected sperm. The study indicates that larvae of olive barb produced from cryopreserved sperm are equally compatible in growth and survival as the larvae produced from fresh sperm. Therefore, cryopreserved sperm can be applied for artificial fertilization of P. sarana to supply quality seed for aquaculture.     

Polyculture of silver barb, Puntius gonionotus (Bleeker), Nile tilapia, Oreochromis niloticus (L.), and common carp, Cyprinus carpio L., in Vietnamese ricefields: feeding ecology and impact on rice and ricefield environment

Rice production, ricefield environment and the feeding ecology offish were studied in an experiment conducted at a rice-fish station in the Mekong Delta, Vietnam. In total, six treatments (three replicates) were investigated: four different polyculture combinations of small sized silver barb, Puntius gonionotus (Bleeker), Nile tilapia, Oreochromis niloticus (L.), and common carp, Cyprinuso carpio L.; one treatment with pre-grown fingerlings; and a control treatment in which no fish were stocked. No insecticides or fungicides were utilized before or during the experiment. Frequent fertilization of the water and a low rice plant biomass during the early vegetative growth phase stimulated the development of phytoplankton and zooplankton. The total weed biomass was low (maximum = 5.3 g dry weight m^?2) and not significantly (P < 0.05) different between the treatments. A major component of the silver barb diet consisted of rice plants and accessible grains. However, the introduction of silver barb only had a significant effect on the number of rice tillers in the ratoon crop and not on the paddy yield. The quantitative differences in the diets of tilapia and common carp were minimal: both species fed mostly on detritus. Ricefields without silver barb produced the highest paddy yield (3120 kg ha^?1). The total yield of introduced fish increased after increasing the stocking density of silver barb from 319.9 to 494.1 kg ha-¹. The highest fish yield (541.8 kg ha-1) was obtained by stocking pre-grown fingerlings in the ricefields.     

Effects of dietary mono‐ and multiprobiotic strains on growth performance,gut bacteria and body composition of Javanese carp (Puntius gonionotus,Bleeker 1850)

This study investigated the effects of dietary mono‐ and multiprobiotic strains of Enterococcus faecalis, Lactobacillus fermentum and Leuconostoc mesenteroides on growth performance, intestinal bacteria and body composition of Javanese carp (Puntius gonionotus). Moreover, the effect of storage temperature on lactic acid bacteria (LAB) viability in the feed was studied. Compared to the control group, significant (P < 0.05) effect on weight gain, feed conversion ratio, specific growth rate and protein efficiency ratio of probiotic fed Javanese carp was observed. Inclusion of individual LAB strains in the diet resulted in significantly improved growth performances of Javanese carps compared to fish fed a diet supplemented LAB mixture. The inclusion level of E. faecalis at 10⁷ colony‐forming unit (cfu) g⁻¹ in the diet revealed higher growth performance compared to other treatments. The population levels of LAB in the fish intestine were significantly increased while the levels of Gram‐negative bacteria were significantly decreased compared to the control. No significant variations were observed in the chemical composition of the carcass by probiotics. The viability of LAB in feed was higher when stored at 4 °C compared to 25 °C. It is suggested that E. faecalis has a greater potential for use in improving Javanese carp culture outcomes.     

Cryopreservation of rainbow trout spermatozoa: The influence of sperm quality,egg quality and extender composition on post-thaw fertility

The presence of 5% to 20% hen's egg yolk in a sucrose-based extender significantly improved post-thaw fertility of cryopreserved rainbow trout (Salmo gairdneri) spermatozoa compared to when the extender without hen's egg yolk was used. However, the degree of increased cryoprotection associated with hen's egg yolk was affected by the quality of the milt. Considerable variation was detected in the performance of various batches of trout eggs used to test post-thaw fertility and the composition of the extender was shown to affect fertilizations differentially with some of the eggs. Despite this variation, the extender containing 10% hen's egg yolk consistently gave high post-thaw fertility in samples of cryopreserved milt (67.3±3.0% S.E.M.) in thirty replicated trials. As such, the method described is reliable for cryopreserving rainbow trout milt and fertilizing small quantities of eggs.