Chilled storage of banana shrimp (Fenneropenaeus merguiensis) spermatophores with supplementation of moringa (Moringa oleifera Lam.) extract

This study was designed to optimize the chilled storage method for banana shrimp (Fenneropenaeus merguiensis) spermatophores and evaluate the potential of moringa (Moringa oleifera Lam.) extract on the reduction in bacterial contaminants during spermatophore preservation due to the uncertainty of the broodstock. Spermatophores were suspended in five extenders: mineral oil, Ringer's solution, phosphate buffer, calcium‐free saline and 0.8% NaCl and stored at 2–4°C. During a 28‐day storage, spermatophores stored in mineral oil showed the highest sperm viability (89.1%) and intact morphology with a slight formation of hardened adhesive matrices. The effect of moringa extract was investigated on chilled spermatophores. Spermatophores were suspended in mineral oil (the control) and mineral oil containing either penicillin–streptomycin (0.1%) or moringa extract (0.1 mg/ml) during a 28‐day storage. Supplementation of moringa extract resulted in a significant increase (p < .05) in sperm survival, compared to the control, and a complete elimination of culturable Vibrio (Vibrio alginolyticus, Vibrio mimicus and Vibrio furnissii), Staphylococcus kloosii, Bacillus macerans, Listeria ivanovii, Corynebacterium paurometabolum and Corynebacterium bovis, in chilled spermatophores. Chilled storage of spermatophores in mineral oil containing moringa extract was a promising technique due to the inhibition of shrimp and human pathogens without the spermicidal effect on banana shrimp sperm.     

Sperm Quality of Pond-Reared and Wild-Caught Penaeus monodon in Thailand1

Sperm quality, as determined by visual examination and by reaction with “egg-water” was not significantly different (P > 0.05) for sperm obtained by electro-ejaculation from ablated or non-ablated pond-reared Penaeus monodon. Nor was sperm quality different between pond-reared and wild-caught prawns. Normal sperm, determined by appearance, ranged from 17.1 to 21.0%, while reactive sperm ranged from 1.5 to 3.0%. There were, however, significant correlations (P < 0.01) between spermatophore weight and prawn weight (r= 0.73, N= 434). Male prawns weighing 4150 g produced on average spermatophores weighing 22.7 mg and containing 0.8 million sperm, while prawns weighing 61-90 g produced on average spermatophores weighing 56.6 mg with 2.5 million sperm. Ablation did not increase spermatophore size or sperm quality, although it significantly increased mortality of ablated males. Male prawns could be re-ejaculated at about weekly intervals with no change in sperm quality. Wild-caught female prawns artificially inseminated with spermatophores from electro-ejaculated males produced normal spawns with 51% average egg fertilization, and 41% nauplii hatch success. Nauplii hatch success following spawning increased from >60% for newly inseminated females to near zero after 30 days post-insemination, indicating spermatophore depletion and/or deteriorated sperm quality during spermatophore storage in the thelycum. The findings of the present study indicate that electro-ejaculation and artificial insemination are relatively simple and practical methods for improving captive reproduction performance of closed-thelycum prawns such as P. monodon, and that pond-reared and wild-caught males produced sperm of similar quality.     

Current status and prospects of farming the giant river prawn Macrobrachium rosenbergii (De Man) in Bangladesh

Giant river prawn (Macrobrachium rosenbergii) farming plays an important role in the economy of Bangladesh. Presently, it is cultured in around 50 000 ha area with total annual production of 23 240 t. Traditional extensive prawn farming has been expanding over the last three decades through the introduction and adoption of improved culture systems, such as culture of prawn‐carps, prawn‐shrimp‐fish and prawn‐fish‐rice as concurrent and rotational systems. Efforts for the development of improved techniques on broodstock management, seed production and rearing and grow‐out of prawn have been made over the last decade. The outcomes are low‐cost feed for broodstock, production of post‐larvae in net cages (hapa), all‐male prawn culture, periphyton based prawn‐tilapia culture, C/N based prawn culture, organic prawn farming, prawn‐mola culture and prawn‐carp‐mola polyculture. Despite the development of culture technologies, a number of challenges for sustainable development of prawn farming need to be overcome to realize the potentials of this promising sector. Good aquaculture practises at all levels and application of measures for quality control and food safety would ensure sustainable development of prawn farming in Bangladesh.     

The effects of dietary HUFA level on flame angelfish (Centropyge loriculus) spawning,egg quality and early larval characteristics

This study was conducted to determine the effects of dietary highly unsaturated fatty acids (HUFA) on flame angelfish (Centropyge loriculus) reproduction, and egg and larval quality. In the experiment, formulated diets containing 1.8, 2.9 or 3.6% n‐3 HUFA were fed to flame angelfish broodstock (n = 4) for 5 months. Mean fecundity (daily egg production), egg fertilization rates and embryo viability were used as indicators of egg quality. In addition, mean egg diameter, oil globule diameter, per cent hatch, larval size at hatch, per cent survival to yolk exhaustion and larval size at yolk exhaustion were recorded for each treatment. Flame angelfish fed the diet containing 3.6% n‐3 HUFA exhibited significantly increased fecundity, fertilization rates and embryo viability than fish that were fed the other two formulated diets. Egg diameter, egg oil globule diameter, larval size at hatch, larval survival to yolk exhaustion and larval size at yolk exhaustion from the 3.6% n‐3 HUFA broodstock treatment group were not significantly different from those derived from the Control broodstock treatment. These data revealed that flame angelfish egg quality could respond rapidly (within weeks) to maternal dietary changes. Results from this study further support that dietary HUFA composition can significantly affect broodstock reproductive performance as well as subsequent performance of eggs and larvae.     

Effect of dietary protein content on growth rate,survival and body composition of juvenile cauque river prawn,Macrobrachium americanum (Bate 1868)

We determined the effect of four concentrations of dietary crude protein, 30.7, 37.2, 41.8 and 46.8% on growth rate, survival and body composition of the juvenile cauque river prawn (Macrobrachium americanum). The prawns were hatched in the laboratory from the spawn of one wild ovigerous female. Prawns consuming 37.2% crude protein reached a final weight of 0.58 g (feed conversion ratio of 2.15), which was significantly better than the other treatments. Survival was 100% in all treatments. Protein content in the diets had no significant effect on whole body proximate composition and amino acid profile. Juveniles consuming the 37.2% crude protein diet grew faster than those fed the other diets. Specific growth rate was adjusted to the two‐slope broken‐line regression analysis model to estimate the optimal protein requirement. In conclusion, these results demonstrated that the 37.2% protein level diet is optimal for juvenile cauque river prawn M. americanum in the experimental conditions of this study.     

Maturation diet based on fatty acid content for male Penaeus monodon (Fabricius) broodstock

The contents of three essential fatty acids, arachidonic acid (AA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA), from wild Penaeus monodon broodstock were evaluated in comparison with natural diet fed P. monodon. Spermatophores of wild male broodstock contained higher levels of AA than those of artificial diet fed males. Polychaetes had higher proportion of AA to EPA and DHA at 5.8:5.5:1 in mud polychaetes followed by 12:7:1 in sand polychaetes, while DHA was a preferential n‐3 highly unsaturated fatty acid (HUFA) in squids and fish. The experimental feed was constructed to simulate the HUFA profile of polychaetes (AA:EPA:DHA as 5:1:1) and then fed to farmed male black tiger prawn broodstock for 1 month. The results exhibited comparable reproductive characteristics to wild male suggesting the possibility of replacing wild males with pond‐reared males. Rearing farmed males in a test unit for a month did not reduce the quality of prawn sperm. Reproductive performance indices (sperm sac weight, total number of sperm, percentage of live sperm, percentage of abnormal sperm) from the males of all treatments were not statistically different except in males fed with pellets. Control (live feeds) and combined diet provided better reproductive performance in pond‐reared males. Analysis of AA, EPA and DHA in reproductive tissues, hepatopancreas and muscle of treated animals in each treatment revealed an accumulation of dietary HUFA into reproductive tissues. No evidence of transfer of HUFA from hepatopancreas or muscle to spermatophore was found.     

Current status and prospects of farming the giant river prawn Macrobrachium rosenbergii (De Man) and the monsoon river prawn Macrobrachium malcolmsonii (H.M. Edwards) in India

Freshwater prawn production in India that includes farming and wild capture of the giant freshwater prawn, Macrobrachium rosenbergii and the monsoon river prawn, M. malcolmsonii has increased steadily since 1999 reaching a peak output of 42 780 t in 2005, but then declined to 6568 t in 2009–2010. Stunted growth and diseases in ponds because of poor seed quality and the broodstock which had been inbred over several generations; pond water quality issues; and increased cost of production on account of feed, labour and the mandatory certification requirements are suggested to be some of the factors leading to the production declines. While majority of the output occurs in Andhra Pradesh, single crop paddy–prawn production systems in the low‐lying fields of Kerala have helped gradual transformation to a sustainable, organic mode of farming of both rice and prawns, suitable for other states of India. Although the trends by June 2011 indicate that the sector is set to a revival, future prospects of freshwater prawn farming in India will also depend on the expansion of whiteleg shrimp Litopenaeus vannamei that was introduced recently in India and provided a more profitable opportunity for farming.     

Effect of vitamin E on spermatophore regeneration and quality of pond‐reared,black tiger shrimp (Penaeus monodon)

The objective of the present work was to evaluate the effects of different levels of vitamin E in feed on the spermatophore regeneration and quality of male Penaeus monodon. The experiment was carried out with the four following treatments: the basal diet no added vitamin E, the diet added 200, 600 and 1,000 mg/kg respectively. Spermatophore regeneration and quality were evaluated by spermatophore weight, sperm count and spermatophore absence rates, which male P. monodon were extruded spermatophore for feeding 20 and 40 days. In the experiment, the weight of the twice regenerated spermatophore of the males added to the vitamin E group was higher than that of the untreated control group. The weight of the first regenerated spermatophore with the addition of 1,000 mg/kg group was the highest and significantly higher than the control group (p < .05), but there was no significant difference among the three groups with different levels of vitamin E. The weight of the second regenerated spermatophore with the addition of 600 mg/kg group was the highest, followed by 1,000 mg/kg group, both of which were higher than the control group and the addition of 200 mg/kg group. Within the same group, the regeneration spermatophore weight showed overall upward trend as the feeding time, twice regenerate experiment spermatophore weight with added to the vitamin E groups were significantly higher than the initial value (p < .05), but three spermatophore weight of male shrimp at the control group had no significant difference. The sperm quantity and the percentage of normal sperm of the twice regenerated spermatophore of the males with added to the vitamin E group was higher than that of the untreated control group, and those of the addition of 200 mg/kg group was significantly higher than that of control group (p < .05). The total number of sperm and the percentage of living sperm of male shrimp in the experimental group decreased with the increase of vitamin E in the feed. Within the same group, the total number of sperm and the percentage of living sperm of male shrimp with added to the vitamin E groups showed overall upward trend as the feeding time and were significantly higher than the initial value (p < .05), but the control group was slightly down and had no significant difference. Comprehensive sperm weight, sperm quantity and living sperm percentage of three indicators, that adding 200 mg/kg of vitamin E in feed could effectively promote the spermatophore regeneration in the male P. monodon and improve the sperm quantity. The experimental results provide a scientific basis for the breeding of P. monodon.     

Growth and economic analysis of freshwater prawn,Macrobrachium rosenbergii (de Man), produced with feeds substituting sunflower cake for fish meal,soya bean meal and mustard oil cake

A feeding trial with Macrobrachium rosenbergii was carried out to test the effectiveness of sunflower cake (SFC) as a replacement for conventional protein sources. Four isonitrogenous (~30% crude protein) and isocaloric (388–402 kcal) diets were formulated. Diet‐1 was a control made with conventional ingredients including fish meal, soya bean meal and mustard oil cake. Diet‐2, Diet‐3 and Diet‐4 were made with 30%, 40% and 50% SFC, respectively, replacing the usual protein sources used in Diet‐1. After 100 days, diets with SFC inclusion showed favourable growth and economic performance trends compared to control diet‐1, but without significant differences (p > .05). Better production (572.89 ± 62.99 kg/ha) was observed with Diet‐4 followed by Diet‐2, Diet‐3 and Diet‐1. Diet‐4 revealed the best feed conversion ratio (2.56 ± 0.24), specific growth rate (1.18 ± 0.05) and protein efficiency ratio (1.14 ± 0.10) among these diets. Whole carcass composition analysis revealed highest crude protein (18.19%) and lipid (4.43%) content in prawns fed Diet‐3 followed by Diet‐4. Highest total income, gross margin and benefit cost ratio (BCR) per hectare from prawns were US$5,759, US$2,623 and 1.84, respectively, found in Diet‐4. This study demonstrates that 30%–50% inclusion of SFC has favourable impacts on productivity and profitability compared with controls, with 50% SFC inclusion as the most cost‐effective diet for prawns without apparent compromise in growth, flesh quality or economics. The performances of the diets suggest that SFC may be a suitable substitute for fishmeal, soya bean meal and mustard oil cake in prawn feed. The effectiveness of higher SFC concentrations should also be investigated.     

In vivo and in vitro protein digestibility and growth performance of animal and no‐conventional plant ingredients for river prawn Macrobrachium americanum (Caridea: Palaemonidae)

The objectives of this study were to evaluate the effects of non‐traditional plant and animal protein for river prawn M. americanum on in vivo versus in vitro digestibility in addition to growth performance. In this study, ten ingredients were used: coconut meal, jackfruit meal, soybean meal, poultry by‐product meal, squid meal and fish meal. In vivo studies were conducted using the zeolite marker method, whereas in vitro digestibility was determined by the pH‐stat method using river prawn hepatopancreas enzymes. According to the findings, river prawns digest animals more efficiently than plant ingredients, with squid and fish meal showing a tendency of higher values. Fish meal (53.66%), squid meal (48.52%) and jackfruit meal (42.21%) were found to have a digestibility greater than 40% in in vivo and in vitro methods, thus suggesting that they are the best ingredients used in this study. This also validates their inclusion in practical diets. The correlation of digestibility of in vivo and in vitro was low (R² = .6749). The highest daily weight gain was recorded in the diets containing jackfruit, coconut and squid. Given that the potential of no‐conventional plant ingredients available regionally was significantly higher, this underscores the need to conduct further research to validate their inclusion in practical diets.     

Egg Water Induced Reaction and Biostain Assay of Sperm from Marine Shrimp Penaeus vannamei: Dietary Effects on Sperm Quality

Reproductive performance was evaluated for sexually mature male Penueus vannamei fed one of four diets: 1) commercial 40% protein shrimp growout diet; 2) 100% squid; 3) 50% squid, 25% bloodworms and 25% Artemia; and 4) starvation. Spermatophores were ejaculated manually from tagged males at stocking and every 12 d thereafter. For each half of the compound spermatophore, weight and sperm count were determined. Four methods were used to assay the sperm quality: 1) gross morphology (GM); 2) trypan blue stain (TB); 3) acridine orange stain (AO); and 4) egg water induced reaction (EW). The reaction induced when P. vannamei sperm are exposed to conspecific egg water is grossly similar to that observed in vivo at the surface of the freshly spawned eggs from naturally mated females. Little correlation was found between spermatophore weight and sperm count. The best correlation was found between GM and AO (r²= 0.859, P < 0.01). Many morphologically abnormal sperm did not stain with TB. For most of the spermatophores evaluated; over 80% of the sperm were morphologically normal, were unstained with TB, and were immediately fluorescing light green when stained with AO. When exposed to egg water, sperm from these apparently good quality spermatophores yielded mixed results. The analysis of dietary effects on spermatophore quality were equivocal. Although a significant decline in body weight, spermatophore weight, and sperm count were observed in the starvation treatment as compared to maturation and squid diets, no significant differences were observed in MY of the sperm quality assays.     

Growth and immune response of giant freshwater prawn Macrobrachium rosenbergii (De Man) postlarvae fed diets containing Chlorella vulgaris (Beijerinck)

A 50‐day growth trial was conducted to evaluate the efficacy of Chlorella vulgaris (Beijerinck) as an ingredient in the diets of giant freshwater prawn Macrobrachium rosenbergii (De Man) postlarvae (PL30). Immune response (total haemocyte count and prophenoloxidase activity) was also assessed by subjecting postlarvae to a challenge test against Aeromonas hydrophila (Chester) for 14 days. Iso‐nitrogenous and iso‐lipidic test diets were prepared using a fishmeal‐based‐positive control diet (D0) and four basal diets with inclusion levels of 2% (D2), 4% (D4), 6% (D6) and 8% (D8) C. vulgaris. Postlarvae of M. rosenbergii were randomly stocked (mean initial body weight of 0.19 ± 0.02 g) in 30‐L tanks in three replicates per dietary treatment for evaluation of growth performance. Another set of postlarvae (mean initial body weight of 1.25 ± 0.02 g) was randomly distributed in 95‐L tanks in three replicates per dietary treatment for the assessment of immune response. Results showed that specific growth rate was significantly higher (P < 0.05) in postlarvae fed D4 and D6. Variations in values for carcass protein, lipid, moisture and ash were also evident. Postlarvae fed diets with Chlorella showed increased prophenol oxidase activity and total haemocyte counts. Moreover, survival rate after challenge with A. hydrophila was significantly increased (P < 0.05). Inclusion of C. vulgaris in diets enhanced immune response and resistance of M. rosenbergii postlarvae against A. hydrophila infection.     

Maternal immunity of tilapia broodstock vaccinated with polyvalent vaccine and resistance of their offspring against Streptococcus agalactiae

This study aimed to examine the use of Streptococcus agalactiae polyvalent vaccine in tilapia broodstock and the effect of maternal immunity and resistance on their offspring against S. agalactiae strain. The broodstock was injected with polyvalent vaccine of S. agalactiae at a dose of 10⁸ CFU per fish at 2nd gonadal maturity until spawning. Challenge test was carried out on the offspring at the 5, 10, 15 and 20 days after hatching using NK₁, N₁₇O, N₁₄G, N₃M, N₄M strain respectively and combination of them. We observed immunological parameters in broodstock, eggs and larvae and relative per cent survival (RPS) of larvae after challenged with pathogenic S. agalactiae. The results showed that the leukocytes, phagocytic activity, respiratory burst, lysozyme activity and antibody levels of vaccinated broodstock had higher level compared with unvaccinated broodstock. The high level of the lysozyme activity, antibody levels and recombination activating gene 1 (RAG1) were also observed in eggs and larvae from vaccinated broodstock. Larvae produced from vaccinated broodstock when challenged with variety strain of pathogenic S. agalactiae had RPS value more than 50% until 20 days after hatching. In conclusion, polyvalent vaccine of S. agalactiae administrated in the broodstock could enhance immunity in the broodstock and protect their offspring from pathogenic S. agalactiae.     

Multiple dietary administrating strategies of water hyacinth (Eichhornia crassipes) on enhancing the immune responses and disease resistance of giant freshwater prawn,Macrobrachium rosenbergii

The different products of Eichhornia crassipes leaves including dried E. crassipes powder (DEP), hot‐water treated E. crassipe (HTE), hot‐water extract of E. crassipe (ECE) and dreg of hot‐water extract of E. crassipe extract (dECE) were produced and incorporated into the diet of the prawn, Macrobrachium rosenbergii, as an immunostimulant. Results showed that prawn fed the HTE‐, ECE‐ and dECE‐containing diets for 4 months had increased total haemocyte count, different haemocyte count, phenoloxidase activity, respiratory bursts, superoxide dismutase activity, glutathione peroxidase activity especially in HTE and ECE treatments. The phagocytic activity and clearance efficiency against Lactococcus garvieae of prawn fed the HTE‐ and ECE‐containing diets were significantly higher than those of prawn fed the control diet at 2–4 months. The relative percentage survival of prawn fed the DEP‐, HTE‐, ECE‐ and dECE‐containing diets for 4 months following 144 h challenging with L. garvieae were 19.0%, 38.1%, 38.1% and 33.3%. It was concluded that E. crassipes leaves containing an active component which was easily extracted by hot water can enhance innate immunity and resistance against pathogen of M. rosenbergii by dietary long‐term administration, and the administration of HTE in the diet was the best strategy due to the availability and convenience.     

Effect of diets on carotenoid content,body coloration,biochemical composition and spermatophore quality in the “red cherry” shrimp Neocaridina davidi (Caridea,Atyidae)

In aquarium industry trade markets, Neocaridina davidi, the “red cherry” shrimp, is one of the most attractive ornamental shrimps due to the striking reddish coloration of females. Five commercial diets formulated for ornamental species were evaluated over a 90‐day period in N. davidi males and females, based on survival, biochemical composition, body weight, carotenoid content and body coloration. Furthermore, histological and histochemical analyses of the male reproductive system were performed to evaluate the spermatophore quality. In all treatments, survival was >80%, and protein content was similar in both female and male shrimps. Lipid content was higher in females and males fed on a diet having the highest lipid level. Carotenoid content was higher and body coloration was brighter in females and males fed on a diet containing the highest carotenoid level. Histological and histochemical analyses of the spermatophore structure and composition showed no differences among diets, suggesting that the five commercial diets are adequate for satisfactory growth and spermatophore quality of N. davidi. The diverse sources and quantities of carotenoids in the highest carotenoid level diet render it more suitable for colour enhancement of this species.     

Sperm production in the red claw crayfish Cherax quadricarinatus (Decapoda, Parastacidae)

The objective of this study is to evaluate the effect of body size, temperature and annual cycle on sperm production in Cherax quadricarinatus. Sperm count and sperm mortality were estimated, the reproductive system was weighted, and macro and microscopical analysis of the testes and vasa deferentia were conducted. Sperm count and weight of the reproductive system are strongly related to male size, in contrast to sperm mortality. The spermatophore structure presented macro and microscopical differences between sizes. Males higher in size have more adherent spermatophores. This species has a reproductive cycle related to sperm production. Sperm count and weight of the vasa deferentia rise in summer, while the weight of the testes increases in winter. During the spring, the sperm cord presents a higher density than in other seasons. The temperature seems to affect sperm production being 27 and 29 °C the best assayed conditions. The present results are relevant information to obtain the best sperm viability selecting male size, season of sampling and the best temperature for the reproductive stock and future assays of spermatophore cryopreservation for this species aquaculture.     

Effects of dietary n‐3 to n‐6 fatty acid ratios on spawning performance and larval quality in tongue sole Cynoglossus semilaevis

This study investigated the effect of n‐3 to n‐6 fatty acid ratios in broodstock diets on reproduction performance, fatty acid composition of eggs and gonads of tongue sole Cynoglossus semilaevis. Broodstock were fed five isonitrogenous and isoenergetic diets for 60 days. The supplemented lipids were prepared by a combination of fish oil and soybean oil inclusion FO (fish oil); FSO1 (fish oil: soybean oil = 7:1); FSO2 (fish oil: soybean oil = 2.2:1); FSO3 (fish oil: soybean oil = 1:1); FSO4 (fish oil: soybean oil = 1:4.3) as lipid sources with different n‐3 to n‐6 fatty acid ratios 10.40, 5.21, 2.81, 1.71 and 0.87. Results showed that relative fecundity, fertilization rate and survival rate of larvae at 7 days posthatching were all higher in broodstock fed FSO1 and FSO2 diet and significantly (P < 0.05) decreased in groups fed FSO3 and FSO4 diets. The best result in starvation tolerance test was obtained in FSO2 diet. The present study suggests that n‐3 and n‐6 PUFA ratio in broodstock diet has a considerable effect on spawning performance, egg and larval quality for C. semilaevis.     

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.     

Effects of replacing dietary fish oil and squid liver oil with vegetable oils on the growth,tissue fatty acid profile and total carotenoids of the giant freshwater prawn, Macrobrachium rosenbergii

A feeding trial was conducted to investigate the complete substitution of either fish oil (FO) or squid liver oil (SLO) with crude palm oil (CPO), canola oil (CO) sunflower oil (SFO) or linseed oil (LO), as the sole added lipid source in diets fed to triplicate groups of giant freshwater prawn, Macrobrachium rosenbergii (initial weight = 0.42 ± 0.01 g) for 6 weeks. Prawns fed the CO or SLO diets showed significantly higher (P < 0.05) specific growth rate than those fed the FO or CPO diets. The feed conversion ratio of the prawns was significantly better when fed the CO diet, compared with the FO, CPO, SFO and LO diets. The muscle eicosapentaenoic acid content of prawns fed the vegetable oil (VO) diets were not significantly different (P > 0.05) from those fed the FO diet, although all VO‐based diets led to a significantly lower docosahexaenoic acid content compared with prawns fed the FO or SLO diet. The whole‐body total carotenoid content was significantly lower for prawns fed the SLO diet compared with prawns on the CO or CPO diets. The successful use of VO instead of marine‐based oils in prawn diets will likely reduce feeding costs associated with M. rosenbergii aquaculture.     

Diet formulation for Macrobrachium rosenbergii (de Man) broodstock based on essential amino acid profile of its eggs

Twelve pelleted diets containing a range of protein and energy levels from 30% protein, 400 kcal gross energy per 100 g diet to 40% protein and 520 kcal gross energy per 100 g diet were formulated. A least-cost linear programming package was applied to determine the optimum inclusion levels of some locally available ingredients. The essential amino acid constraints applied to the diets were based on the essential amino acid content of the eggs of broodstock Macrobrachium rosenbergii (de Man). The study was undertaken using 13 one-tonne capacity fibreglass tanks, each with a water recycling system. Each tank was partitioned into three equal compartments by nylon netting; each compartment was stocked with six female and one male prawn which were fed with the various diets at 2% body weight daily split into three feedings at 0800, 1200 and 1800 h. The results indicated that prawn fed the 40% protein diet with an energy level of 400 kcal per 100 g diet attained the highest fecundity, producing 1355 eggs per gram body weight, followed by those fed P40 with a gross energy level of 440 kcal 100 g^-1 (1354 eggs per gram body weight), and prawns fed control diet (30% protein with an energy level of 442 kcal 100 g diet^-1) attained the lowest fecundity (1080 eggs per gram body weight). However, statistically, no significant differences in fecundity were observed among prawn fed 35% protein diet with an energy level of 473 kcal 100 g^-1, 40 with energy levels of 400 or 440 kcal 100 g diet^-1. The essential amino acids index (EAAI) were calculated and were found to be a possible method of evaluating the broodstock diet of M. rosenbergii as a higher index indicates higher egg production. Therefore, a diet containing 40% protein, with an energy level of 400 kcal 100 g diet^-1 is recommended as a broodstock feed for M. rosenbergii in view of its superior performance and cost.