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.

Species interaction between carp species in polyculture system

A three months experiment was conducted to study the species interactions of two carp species in polyculture system under supplemental feeding. Four species of fishes such as silver carp (Hypophthalmichthys molitrix), mrigal (Cirrhina cirrhosus), catla (Catla catla) and common carp (Cyprinus carpio) were cultured in four different combinations each containing two species. The combination of silver carp and mrigal in treatment 1, and silver carp and common carp in treatment 2 resulted better growth and production than other two treatments of different combinations of catla and common carp, and catla and mrigal.

Effects of iso-nitrogenous fertilizers as nutrient sources on carp polyculture in Bangladesh

A 120-day long experiment was conducted to find out the effects of urea plus triple super phosphate (UT), cow manure (CM) and poultry manure (PM) having iso-nil:rogen content on pond productivity and fish yield. Three fertilizer treatments, with three replicates each, were randomly assigned into nine earthen ponds of 100 m2 each. The stocking fish were rohu (Labeo ruhita), catla ( Catla catla) and mrigal ( Cirrhinus mrigala) in each treatment pond at the rate of 10000/ha with the ratio of 1:1:1. All ponds were fertilized fortnightly at the rate of 125 kg/ha urea plus 100 kg TSP/ha, 7000 kg/ha cow manure and 3500 kg/ha poultry manure for the treatment of UT, CM and PM, respectively, having an iso-nitrogen content of 56 kg in each. Though the physicochemical water quality parameters were more or less similar in all treatment ponds, the chlorophyll-a content and abundance of total plankton were significantly higher (P < 0.05) in the ponds receiving the treatment PM. Final growth as well as per unit production of fish was significantly higher (pcarp polyculture system under prevailing conditions.

Bacterial load in pond water and different organs of a Indian major carp Cirrhinus mrigala Ham.

During the study period (August, 1993 to July,l994) the mean bacterial load in surface water was found to vary from 1.39 xl05 (July'94) to 3.llxl07CFU/ml (September'93), while that of botrom water ranged from l.Olxl06 (May'94) to 5.90xl07CFU/ml (October '93). The mean total number of bacterial load in body slime, liver and kidney was found to vary from 0.58xl03 (July'94) to 2.37xl07CFU/g (March'94),from 0.22xl03(July'94)to 9.64xl06 CFU/g (March'94) from O.l5xl03 (July'94) to 9.36xl06 CFU/g (March'94), respectively. Bacterial load in slime was significantly correlated with bacterial load in liver, bacterial load in slime was significantly correlated with bacterial load in kidney and bacterial load in liver was significantly correlated with bacterial load in kidney.

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.

Relationship between total length and mouth gap of some commercially important carp fry

The estimated regression equation for total length and mouth gape computed were Log TL = Log 0.23 + 0.663 log MG (vertically) (r = 0.960) and Log TL = Log 0.08 + 0.686 log MG (horizontally) (r = 0.949). In case of rohu average total length from 11350 mm to 23775 mm and mouth gape 805 um to 1225 um (vertically) and 700 um to 1110 um (horizontally) between the first day of mouth opening up to 15 days. The regression equation for total length and mouth gap were Log TL = Log 0.20 + 0.660 log MG (vertically) (r = 0.935) and Log TL = Log 0.02 + 0698 log MG (horizontally) ( r = 0.907). In case of silver carp average total length from 12800 ,urn to 33555 um and mouth gape 690 um to 1210 um (vertically) and 615 um to 1115 um (horizontally) between the first day of mouth opening up to 15 days. The regression equation for total length and mouth gape were Log TL = Log 0.36 + 0.596 log MG (vertically) (r = 0.936) and Log TL = Log 0.26 + 0.607 log MG (horizontally) (r = 0.891). The relationship between total length and mouth gape (vertically and horizontally) of the studied fry were found to be linear and highly significant.

Effect of carp PG doses on induced breeding of Shing, Heteropneustes fossilis (Bloch)

Five hormone doses viz. 25, 50, 75, 100, and 125 mg of carp PG/kg of body weight of the recipient fish were tested and they were designated as T1 T2, T3, T4, and T5 respectively. Significantly higher fertilization (98%) and hatching rates (38%) were obtained from T3 (75 mg of carp PG extract/kg body weight). While T4 (100 mg of carp PG extract/kg body weight) and T5 (125 mg of carp PG extract/kg body weight) gave the highest (90%) ovulation rate. In June and July the highest fertilization rate of 96 and 96.4% respectively and hatching rate 42.5 and 48.7% respectively were obtained. In over all consideration carp PG extract at a dose of 75 mg/kg body weight appears to be the suitable dose for induced breeding of H. fossilis and June and July are the suitable time for its induced breeding.

Effect of feeding frequency on the growth of common carp (Cyprinus carpio L.) fry

A laboratory-feeding trail was conducted for 45 days with fry of common carp Cyprinus carpio L. (0.45±0.03g) in aquaria in a static indoor fish rearing system. The fry were fed on a pelleted diet containing 33% crude protein having fishmeal as major protein source. The fish fry in five treatments A, B, C, D, and E, each with two replicates were fed on 5% daily ration divided into different feeding frequencies of 2, 3, 4, 5 and 6 times a day respectively in order to observe the growth performance. Each replicate contained 15 fry having total initial weight of 6.87±0.31g. At the end of the feeding trial, significantly different and higher (p<0.05) growth response was observed in treatment C having a feeding frequencies of 4 times a day. Significantly the highest and the lowest percent growth of 334.30 and 218.91% were observed in fish fed on the diet (Treatment C) with 4 times and (Treatment A) 2 times feeding frequencies per day, respectively. Food conversion ratio (FCR) of 1.78 was significantly higher (pcarpio was the best at 4 times feeding in a day using 33% dietary protein containing fish meal as major protein source.

Development of canned products from Indian major carp, catla (Catla catla)

Pieces of catla fish (Catla catla, a major carp of lndian subcontinent) of length 10 to 11 cm and thickness 1.0 to 1.5cm were precooked by two methods; steam precooking and frying. The precooked pieces were packed_in No.1 tall can (30lx411) with various hot filling media like oil, brine, tomato sauce and curry. Cans were seamed by a hand seamer and retorted at 117.2° C (12 psi) for 90 minutes. All the canned products had satisfactory cut-out, biochemical and organoleptic characteristics. Steam precooked canned products had moisture content of 65.6 to 74%, protein content of 20.8 to 22%, fat content of 1.1 to 6.6% and ash content of 2.1 to 2.5%; whereas fried canned products had moisture content of 65.4 to 68.2%, protein content of 21.3 to 22%, fat content of 7 to 10.2% and ash content of 2.1 to 2.7% on wet wt. basis. Salt content in steam precooked and fried canned products varied from 1.2 to 1.9% and 2.0 to 2.5% respectively. All the canned products were organoleptically good. However, degree of preference varied for different products. Canned fried catla in curry was the best product among all types of packs. Among the precooking methods, frying was more efficient than steam precooking in controlling the amount of exudate to a desirable limit in canned products. However, crispness, the characteristic quality for a fried fish, was lost during retorting. There was no change in quality characteristics during a storage period of 3 months at ambient temperature (32±2°C).

Effects of iso-nitrogenous and iso-phosphorus fertilizers as nutrient sources on carp polyculture in Bangladesh

A 120 day long experiment was conducted to find out the effects of cow manure with urea and triple super phosphate (CUT), poultry manure with urea and triple super phosphate (PUT) and cow manure with poultry manure (CP) having similar quantities of nitrogen and phosphorus on pond productivity and fish yield. The stocking fish were rohu (Labeo rohita), catla ( Catla catla) and mrigal ( Cirrhinus mrigala) in each treatment pond at the rate of 10000/ha. All ponds were fertilized fortnightly at the rate of 4000 kg/ha cow manure with 62 kg/ha urea and 65 kg/ha TSP, 2700 kg/ha poultry manure with 62 kg/ha urea and 16 kg/ha TSP, and 4000kg/ha cow manure with 2700 kg/ha poultry manure for the treatment CUT, PUT and CP respectively. Each treatment contained an iso-nitrogen and iso-phosphorus of 56 kg and 46 kg respectively. Though the physico-chemical parameters were more or less similar in all ponds, the chlorophyll-a content and abundance of total plankton were significantly higher (P< 0.05) in the ponds receiving the fertilizer treatment of PUT than those of other treatments. Final growth as well as per unit production of fish of treatment PUT (1773 kg/ha) was significantly higher (P< 0.05) than that of treatment CP (1528 kg/ha) followed by that of treatment CUT (1336 kg/ha). The over all results showed that poultry manure with urea and triple super phosphate proved to be superior to cow manure with urea and triple super phosphate, and poultry manure with cow manure, even when nitrogen and phosphorus content was similar, in carp polyculture system under prevailing conditions.

Effect of stocking density on the production of common carp (Cyprinus carpio Lin.) in cages at Kaptai lake, Bangladesh

Growth, survival and food conversion ratio (FCR) of Cyprinus carpio in cages at different stocking density were studied in Kaptai lake. Fingerlings of common carp (12.47 ± 4.29 g) were stocked at 25/m2 , 50/m2 and 75/m2 each with replication in six floating net cages each of Sm x Sm x 4m and reared for 240 days. The growth rate was inversely related to the stocking density with the mean weights of 325.5 ± 11.74 g, 268.99± 8.44 g and 167.0 ± 11.66 g at low, medium and high densities, respectively. At harvest, standing crop biomass averaged 7 .82, 12.83 and 11.58 kg with the survival of 98.9%, 97.6% and 94.4% and food conversion ratios of 4.51, 3.82 and 4.21 for the above three densities, respectively. Weight gain and production at density 25 fish/m 2 were significantly different (p< 0.01) from other two densities. Water quality was not affected in the cages having different stocking densities.

An observation of performances in carp-prawn polyculture in respect of fingerlings size and water area

Performances of fingerling size and water area was studied. Seven species of fish included 25% of Hypophthalmichthys molitrix, 15% of Gada catla, 20% of Labeo rohita, 10% of Cirrhinus mrigala, 10% of Ctenophyringodon idella and 20% of Macrobrachium rosenbergii were stocked at the rate of 8,000 individual/ha along Barbodes gonionoms at 6.0g weight size with a stocking density of 2,000 indl./ ha. Survival of all species was in the range of 39.9 to 96.3% except M. rosenbergii The highest survival was obtained from L. rohita (96.3%) followed by C mrigala (96.0 %)., B. gonionotus (92.3%), C idella (90.6%), H. molitrix (84.0%), C cada (81.7%) respectively. While the survival of M. rosenbergiiwas 13.1%. The highest production of 4912 kg/ha/yr was obtained from the bigger ponds (0.4 ha) stocked with 20-32g size of fingerlings, while lowest production (3123 kg/ha/yr) from small pond (0.1 ha) stocked with 3-10 g fingerlings. Economic analysis shows the highest net profit in bigger ponds stocked with bigger fish seed.

Influence of formulated diets with varying protein levels on growth and ammonia excretion in the fry of Indian major carp, Cirrhina mrigala

The influence of formulated isocaloric diets of different protein levels (30, 35, 40, 45 and 50%) on the growth and ammonia excretion of the Indian major carp Cirrhina mrigala fry was studied for a rearing period of four weeks in the laboratory. Fishmeal, groundnut oilcake and silkworm pupae formed the source of protein in all the diets. As the dietary protein level increased from 30 to 40%, the growth and conversion efficiency increased significantly. Further increase in the protein level resulted in decrease in growth and conversion efficiency. Growth rate, weight gain (%), and gross and net feed conversion efficiencies were maximum at 40% dietary protein level. Ammonia excretion was directly proportional to the level of protein in the diet.

Recovery, growth and carcass composition following periods of restricted ration and full feeding in indian major carp, Rohu (Labeo rohita H.)

A 90-day experiment was conducted to determine the effect of restricted ration and full feeding on the recovery growth and carcass compositions of fingerlings (average weight - 20.74 ± 0.13 g) of rohu, Labeo rohita (H.). Rohu fingerlings procured from a local fish breeder were fed with commercial pelleted feed (27% crude protein) during the two-week acclimatization in the laboratory condition. Experimental pelleted diet (30% crude protein) was prepared and the control group (T sub(CFR)) was fed at 3% of body weight for the 90-day trial period. The experimental group T sub(1FR) was fed for three days at 1% of body weight and the next three days at 3% of body weight, T sub(2FR) was fed for seven days at 1% of body weight and the next seven days at 3% of body weight, T sub(3FR) was fed for 15 days at l% of body weight and the 15 days at 3% of body weight and T sub(4FR) was fed for 25 days at 1% of body weight and the next 25 days at 3% of body weight, alternating between 1 and 3% for the specified period during the 90-day trial period. Daily rations were divided into two equal meals per day at 09.00 and 16.00 hours. Average percent survival rate of rohu during the 90-day trial period was more than 90. Percent live weight gain (98.90 ± 0.34, 113.0 ± 5.93, 125.71 ± 11.01 and 141.90 ± 2.89), specific growth rate (1.53 ± 0.01 1.68 ± 0.06, 1.80 ± 0.10 and 1.96 ± 0.02%/d) and absolute growth rate (1.33 ± 0.13, 1.38 ± 0.07, 1.39 ± 0.04 and 1.44 ± 0.07g/d) of the experimental groups (T sub(1FR), T sub(2FR), T sub(3FR) and T sub(4FR) respectively) increased with the advancement of the experiment in comparison to those in control, T sub(CFR) (90.92 ± 5.81%, 1.44 ± 0.07%/d and 1.34 ± 0.20g/d, respectively) and were proportionately correlated with the degree of deprivation probably through the mechanism of increased feed intake (hyperphagia), feed efficiency ratio or gross growth efficiency, protein efficiency ratio and the superior feed conversion ratio reflecting in better performance index. The body length and muscle composition of fish indicated that recovery growth happened due to protein growth but certainly not due to fat deposition in the gut. Feeding at 1 and 3% of body weight alternating over a period of 25 days might economize the culture operation of rohu.

Effect of sublethal concentration of artificial seawater on oxygen uptake and opercular frequency in an Indian major carp, Labeo rohita (Ham.)

The present study reports the effect of artificial seawater on oxygen uptake and opercular frequency in an Indian major carp, Labeo rohita. Whereas a control fish of 7.34 g average body weight consumed 1.538 ml O sub(2.) hˉ¹, the 24h and 96h exposed fish of the same body weight consumed 1.07 4 and 0.897 ml O sub(2.) hˉ¹, respectively. The oxygen uptake per unit body weight under controlled condition was 0.219 ml. gˉ¹. hˉ¹, whereas in 24h and 96h exposed fish, it was 0.152 and 0.124 ml. gˉ¹, hˉ¹, respectively. The change in opercular movement in 24h exposed fish was 7.67% higher, whereas in 96h exposed fish, it was 22.43% higher as compared to the control one. All changes are highly significant (p<0.001).