Low-cost diet for monoculture of giant freshwater prawn (Macrobrachium rosenbergii de Man) in Bangladesh

An experiment was conducted for 3 months in 12 experimental ponds, each of 30 m², with a view to develop a low‐cost diet for monoculture of Macrobrachium rosenbergii in ponds. Three experimental diets (30% protein) were formulated using fish meal, meat and bone meal, mustard oilcake, sesame meal and rice bran in different combinations partially replacing fish meal by meat and bone meal and sesame meal and assigned to treatments T₁, T₂ and T₃ respectively. A commercial golda feed (Starter‐III) was assigned to T₄ (reference diet). Each treatment had three replicates. Juveniles of M. rosenbergii (2.90±0.21 g) were stocked at the rate of 40 000 ha⁻¹. Prawns were fed three times daily at the rate of 10% and 5% of their body weight at the beginning and for the last 2 months respectively. The ponds were provided with aeration during the night using air pumps. The ranges of water quality parameters recorded in different ponds were: temperature 28.9–32.5°C, dissolved oxygen 5.1–8.1 mg L⁻¹ and pH 6.4–7.7. The results showed that the weight gain of prawns fed diet 1 was significantly higher (P<0.05) than those fed diets 2 and 3, but was not significantly different from those fed diet 4 (reference diet). The feed conversion ratio (FCR) values of diets ranged between 2.21 and 2.96 with diets 1 and 4 showing significantly lower (P<0.05) FCR values. The survivals (%) ranged between 68% and 78% with prawns fed diets 1 and 4 showing significantly higher survival. The production of prawn ranged between 921 and 1428 kg ha⁻¹ and diet 1 resulted in a significantly high (P<0.05) production. A simple economic analysis showed that diet 1 generated the maximum net profit of Tk 159 178 ha⁻¹. The results of the study showed that a diet containing 20% fish meal, 10% meat and bone meal, 15% mustard oilcake, 15% sesame meal, 35% rice bran, 4% molasses and 1% vitamin–mineral premixes may be recommended to the farmers for monoculture of M. rosenbergii in ponds.     

Effect of Bacillus subtilis on growth development and survival of larvae Macrobrachium rosenbergii (de Man)

A feeding experiment was conducted to investigate the effect of Bacillus subtilis bacterium, on larval growth and development rate of Macrobrachium rosenbergii (de Man) during February 28 to April 8, 2005 in University Putra Malaysia hatchery. Newly hatched larvae of M. rosenbergii were reared with two dietary treatments consisting of newly hatched Artemia salina nauplii with B. subtilis (10⁸ cells ml⁻¹), and newly hatched A. salina nauplii without B. subtilis carried out in triplicate in 60‐L aquarium (50 L⁻¹). After trial, the larvae that fed B. subtilis‐treated Artemia naupli were found to have higher survival and a faster rate of metamorphosis than larvae that were fed with nontreated Artemia naupli. There were significant differences between B. subtilis‐treated Artemia naupli and nontreated Artemia diet in larval growth and development rate of metamorphosis (P < 0.05). Larval survival after 40 days was significantly greater (P < 0.05) in the B. subtilis‐treated groups (55.3 ± 1.02) compared with the nontreated groups (36.2 ± 5.02%).     

Optimization of stocking density of freshwater prawn Macrobrachium rosenbergii (de Man) in carp polyculture in Bangladesh

A study was conducted to optimize stocking density of freshwater prawn, Macrobrachium rosenbergii, in carp polyculture for 3 months in 10 experimental ponds of 80 m². Five stocking densities of prawn, 2500, 5000, 7500, 10 000 and 12 500 ha^?1, were assigned to treatments T₁, T₂, T₃, T₄ and T₅ respectively. The densities of catla, Catla catla, rohu, Labeo rohita and silver carp, Hypophthalmicthys molitrix, were 2500, 5000, and 2500 ha^?1, respectively, in each treatment. Each treatment had two replicate ponds. The mean initial weights of prawn, catla, rohu and silver carp were 1.1±0.02, 8.28±0.1, 25.2±1.1 and 36.32±1.2 g respectively. A pelleted diet containing 30% protein was prepared using fish meal, meat and bone meal, mustard oilcake, rice bran, wheat bran and molasses, and was fed twice daily at a rate of 5% of fish biomass. Water quality parameters were measured fortnightly and the ranges of temperature, pH and dissolved oxygen were 27.5–1.3°C, 6.9–8.6 and 4.5–8.6 mg L^?1 respectively. Feed conversion ratios ranged from 2.05 to 2.20 among the treatments. Per cent survival (%) of prawns ranged from 72% to 78%, while it varied from 80% to 93%, 90% to 95% and 90% to 92% for catla, rohu and silver carp respectively. The results showed that there were no significant differences among the weight gains of prawn and carp in different treatments. However, the overall total production of prawn and fish together was significantly (P<0.05) higher in T₃ and T₄ compared with other treatments. The total production for 3 months ranged between 2618 and 2916 kg ha^?1. The production of prawn was significantly higher (361.3 kg ha^?1) in T₅ with a highest stocking density of 12 500 prawn ha^?1. Although there was no significant difference (P>0.05) between the total production of prawn and fish together in T₃ and T₄, the highest net profit (Tk. 69 006 ha^?1) was obtained in T₄. Therefore, from the result of the study it may be concluded that a stocking ratio of 4:1:2:1 of prawn:catla:rohu:silver carp at a total density of 20 000 ha^?1 may be recommended for prawn–carp polyculture in ponds.     

Long-term cryopreservation of spermatophore of the giant freshwater prawn, Macrobrachium rosenbergii (de Man)

Long‐term cryopreservation of the giant freshwater prawn, Macrobrachium rosenbergii, spermatophores using glycerol (Gly) and ethylene glycol (EG) as cryoprotective agents (CPAs) was studied. The tolerance of sperm to cryopreservation was evaluated on the basis of sperm survival and fertilizing ability. The survival of the sperm was determined by trypan blue staining, while the fertilizing ability was assessed from artificial insemination of the cryopreserved spermatophores. The rates of embryo survival on day 5 after spawning and of spermatophores capable of producing embryos survived to hatching were determined. Storage of spermatophores at ?20°C without CPA for a short period of up of 1–5 days decreased the sperm survival significantly and did not preserve fertilizing ability. Preservation at ?20°C in the presence of 10% or 20% Gly or of 10% or 20% EG offered a simple and efficient short‐term storage up to 10 days. For a long‐term storage, cryopreservation in the presence of 20% EG at ?196°C was more efficient than at ?20°C. High sperm survival rates and high fertilizing ability were recorded from those cryopreserved at ?196°C for up to 150 days. High sperm survival rates with moderate levels of fertilizing ability were obtained from those cryopreserved at ?20°C for not more than 30 days. The results indicate that preservation at ?196°C with 20% EG is a suitable procedure for long‐term storage of the giant freshwater prawn spermatophores.     

On the breeding behaviour and reproduction of the freshwater prawn,Macrobrachium rosenbergii (de Man 1879) (Decapoda‐Crustacea) under laboratory conditions

The giant freshwater prawn, Macrobrachium rosenbergii is one of the most common decapod species, and now getting more attention from the aquaculturists world wide due to its high market demand. It is commercially important because of its size as well as its eating flesh qualities. The breeding behaviour, reproduction and hatching of this species were observed for about 8 months during 2009. Juveniles (0.55 ± 0.177 g total weight and 2.7 ± 0.12 cm, total length) were reared in rounded fibre glass tanks (1.3 m, diameter). Pre‐mating moult occurs in prawns once the ovaries ripen in their carapace cavity so as to transform the prawn into berried stage. Recorded incubation period ranged from 18 to 24 days. The number of eggs ranged from 2050 to 150 500 and the fecundity ranged from 435.2 to 3849.1 eggs. Number of hatched larvae ranged from 1825 to 123 410 larvae for females of 4.71 to 39.1 g respectively. The number of eggs carried by female prawn was directly proportional to its body weight (no. of eggs = 3441.3 wt. of female – 32 292, r² = 0.819). (fecundity = 82.066 wt. of female ? 235.04, r² = 0.7779; fecundity = 317.86 length of female ? 2651, r² = 0.833). Hatching rate ranged from 65% to 91%, and there was a correlation between number of hatched larvae and size of female parent (no. of larvae = 10 369 length of female – 102 965, r² = 0.8159; no. of larvae = 2792.9 wt. of female – 26 268, r² = 0.829). This study can greatly help in the management strategies of prawn hatcheries and improve its hatching technology.     

Bacterial microflora associated with farmed freshwater prawn Macrobrachium rosenbergii (de Man) and the aquaculture environment

The initial counts of bacteria associated with farmed giant freshwater prawn (Macrobrachium rosenbergii de Man) as well as with the water and sediment from two farms located at Kottayam district in Kerala (India) were determined. A total of 367 randomly selected isolates were characterized and identified. Prawn samples yielded mean microbiological counts of 4.92 log CFU g^?1 of shell with muscle and 7.78 log CFU g^?1 of intestine at 30°C. Bacterial numbers in the intestine of prawn were much higher than those in the pond water. Motile and non‐motile aerobic Gram‐negative bacteria together with Enterobacteriaceae accounted for 60–70% of the mesophiles isolated from water and prawn. However, the microorganisms isolated in significant numbers from shell with muscle and intestine of prawn were not recovered from the growing waters. Among Gram‐positive bacteria, Micrococcus, Bacillus, coryneforms, and Arthrobacter were found. Faecal coliforms and enterococci were found in significant numbers in M. rosenbergii. The rearing practices such as feeding and pond fertilization could have influenced the microflora in prawn. The commensal microflora of freshwater prawn included opportunistic pathogens such as Aeromonas spp. and Streptococcus spp. Storage of prawn at elevated temperatures can permit their survival and growth leading to quality loss. Care should be exercised during handling and processing to prevent contamination of edible meat.     

Characterization and pathogenicity studies of Vibrio parahaemolyticus isolated from diseased freshwater prawn, Macrobrachium rosenbergii (de Man)

Vibrio parahaemolyticus, an important pathogen of freshwater prawn, Macrobrachium rosenbergii causes high mortality. Four strains of V. parahaemolyticus were isolated from the gill, haepatopancreas, hemolymph and carapace of diseased prawn. The species were characterized based on biochemical and serological tests. On performing an antibiotic assay, these strains were found to be highly susceptible to cephalaxin, tetracycline and erythomycin. During the in vitro pathogenicity test, all the strains were found to be positive to the Congo red binding assay and to be haemolytic in nature, whereas an in vivo pathogenicity test revealed that 2 × 10⁹ CFUs mL^?1of bacteria induced disease symptoms such as black colouration on the carapace, red discolouration of the exoskeleton and loss of appendages within 6 days and 80% mortality. In histopathological studies, a prominent necrosis was seen in gill lamellae, and branchial arches were thickened at places due to hyperplasia and haemocytic infiltration. Hepatopancreatic tissue showed dilation of tubules, vacuolation of hepatocytes and marked necrosis in acinar cells. Development of an immunological technique for the detection and screening of V. parahaemolyticus infection and its treatment is highly important in freshwater prawn aquaculture.     

Production of recombinant capsid protein of Macrobrachium rosenbergii nodavirus (r‐MCP43) of giant freshwater prawn,M. rosenbergii (de Man) for immunological diagnostic methods

White tail disease (WTD) caused by Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV) is a serious problem in prawn hatcheries. The gene for capsid protein of MrNV (MCP43) was cloned into pRSET B expression vector. The MCP43 protein was expressed as a protein with a 6‐histidine tag in Escherichia coli GJ1158 with NaCl induction. This recombinant protein, which was used to raise the antiserum in rabbits, recognized capsid protein in different WTD‐infected post‐larvae and adult prawn. Various immunological methods such as Western blot, dot blot and ELISA techniques were employed to detect MrNV in infected samples using the antiserum raised against recombinant MCP43 of MrNV. The dot blot assay using anti‐rMCP43 was found to be capable of detecting MrNV in WTD‐infected post‐larvae as early as at 24 h post‐infection. The antiserum raised against r‐MCP43 could detect the MrNV in the infected samples at the level of 100 pg of total protein. The capsid protein of MrNV estimated by ELISA using anti‐rMCP43 and pure r‐MCP43 as a standard was found to increase gradually during the course of infection from 24 h p.i. to moribund stage. The results of immunological diagnostic methods employed in this study were compared with that of RT‐PCR to test the efficiency of antiserum raised against r‐MCP43 for the detection of MrNV. The Western blot, dot blot and ELISA detected all MrNV‐positive coded samples as detected by RT‐PCR.     

Genetic diversity of cultured and wild populations of the giant freshwater prawn Macrobrachium rosenbergii (de Man, 1879) based on microsatellite analysis

Freshwater prawn (Macrobrachium rosenbergii) culture in the Western Hemisphere is primarily, if not entirely, derived from 36 individual prawns originally introduced to Hawaii from Malaysia in 1965 and 1966. Little information is available regarding genetic variation within and among cultured prawn stocks worldwide. The goal of the current study was to characterize genetic diversity in various prawn populations with emphasis on those cultured in North America. Five microsatellite loci were screened to estimate genetic diversity in two wild (Myanmar and India‐wild) and seven cultured (Hawaii‐1, Hawaii‐2, India‐cultured, Israel, Kentucky, Mississippi and Texas) populations. Average allelic richness ranged from 3.96 (Israel) to 20.45 (Myanmar). Average expected heterozygosity ranged from 0.580 (Israel) to 0.935 (Myanmar). Many of the cultured populations exhibited reduced genetic diversity when compared with the Myanmar and the India‐cultured populations. Significant deficiency in heterozygotes was detected in the India‐cultured, Mississippi and Kentucky populations (overall F_is estimated of 0.053, 0.067 and 0.108 respectively) reflecting moderate levels of inbreeding. Overall estimate of fixation index (F_st = 0.1569) revealed moderately high levels of differentiation among the populations. Outcome of this study provide a baseline assessment of genetic diversity in some available strains that will be useful for the development of breeding programmes.     

Management strategies associating batch-graded and size-graded postlarvae can reduce heterogeneous individual growth in Macrobrachium rosenbergii (de Man)

Two sets of adaptive trials were performed to determine the effects of size grading and batch grading on size heterogeneity in cultured Macrobrachium rosenbergii raised in the Coconut Garden channels of Kuttanad, Southern India. In the first set of trials, postlarvae were batch graded on the basis of their hatching order, segregated and grown separately as first‐hatched and second‐hatched groups. In the second set of trials, postlarvae were size graded as jumpers and laggards and were grown in separate channels. The average weight attained by prawns after 10 months of culture was highest for jumpers (83.11 g) and lowest for the prawns from the first‐hatched group (43.76 g). The percentage of males was highest in the population of jumpers (58.23%). Highest production was recorded in the channel stocked with postlarvae from the second‐hatched group (103.4 kg ha^?1) and lowest production was obtained from the first‐hatched group (63.74 kg ha^?1). The proportions of undesirable small males were highest among laggards and the first‐hatched group: 24.8% and 15.1% respectively. The level of heterozygosity within morphotypes was also high in these groups. Jumpers attained good growth by the end of culture but, because of their low survival rate, this approach was not economically feasible. However, higher production and survival in the second‐hatched group improved economic viability. Thus, for better results, stocking with later‐hatched groups would be more appropriate than stocking with the first‐hatched group.     

A sandwich enzyme linked immunosorbent assay (S-ELISA) for detection of MrNV in the giant freshwater prawn,Macrobrachium rosenbergii (de Man)

A sandwich enzyme-linked immunosorbent assay (S-ELISA) was developed to improve diagnosis of white tail disease of the giant freshwater prawn, Macrobrachium rosenbergii, caused by the nodavirus, MrNV. Polyclonal antibodies were produced by immunization of Balb/C mice using a purified suspension of the virus and IgG anti-MrNV were purified from ascitic fluid. A sandwich method was successfully developed, coating first with unlabelled antibody and detecting trapped antigens with a second biotinylated antibody. Reaction was demonstrated using an avidin-peroxidase conjugate. Tissue extracts from M. rosenbergii infected with MrNV or purified viral extracts (control) were successfully identified in an individual ELISA, thus confirming the validity of the method. This S-ELISA should be the technique of choice for epidemiological studies of this disease and is a rapid and inexpensive assay with high specificity and sensitivity.     

Growth and digestive enzymes of Macrobrachium rosenbergii juveniles: effect of different stocktypes and dietary protein levels under a similar culture environment

A feeding trial was conducted to study the effect of dietary protein (DP) levels on the growth and digestive enzyme activities of different wild stocks of Macrobrachium rosenbergii juveniles. Wild juveniles of M. rosenbergii were collected from the west coast of India, Gujarat (G), Maharashtra (M) and from the east coast of India, Andhra Pradesh (A), and raised in culture ponds of 200 m² at 1 juvenile m^?2. All the animals were tagged individually with Elastomer tags of a particular colour assigned to their respective stock and acclimatized for 7 days before being released into the pond at a ratio of 70:65:65 (A:M:G). Each of the two feeds, the first with 27% DP, termed the suboptimum level (S), and the second 32% DP, termed the optimum level (O), was fed in duplicate ponds at 6% of the body mass for the first 30 days and 4% for the last 30 days. The average weight of stocked prawn, respectively, in O DP and S DP fed ponds was 0.90 ± 0.04 and 1.06 ± 0.08 g for the G stock, 0.80 ± 0.07 and 1.01 ± 0.1 g for the M stock and 3.06 ± 0.13 and 3.10 ± 0.23 g for the A stock. Both the protein level and the stock type had a significant (P<0.05) effect on the weight gain% of the prawn. There was an approximate 95% change in weight gain with a DP change. Similarly, G and M stocks exhibited significantly higher (P<0.05) growth rates of approximately 90% than the A stock, although no difference was noted between the G and the M stocks. However, for protein × stock (interaction) levels, there was no significant difference (P>0.05) among the groups. Although insignificant, the survival rates among the different stocks varied from 56% to 77%. Optimum protein level showed a significant increase (P<0.05) in the specific growth rate (SGR). Feed conversion ratio, feed efficiency ratio, protein efficiency ratio and net protein utilization were not affected either due to DP, stock type or their interaction. The O × A group exhibited the maximum variation in body weight. Digestive enzyme activities were similar in all the groups, but enzymes for phospho‐monoesterase were significantly higher (P<0.05) at O DP. Both the G and the M stock showed a significantly higher (P<0.05) alkaline phosphatase activity while acid phosphatase activity was significantly higher (P<0.05) in the M stock only. Overall, the G and M stocks showed higher growth responses compared with the A stock.     

The effects of densities and feed types on the production of Macrobrachium rosenbergii (de Man) in the rotational rice–prawn system

Postlarvae (PL) of Macrobrachium rosenbergii (de Man 1879) were stocked at 1, 2, 3 and 4 PL m^?2. Prawns were fed only pellets (P) and a combination of pellets and snail meat (PS) with three replications in 100 m² plots in the rotational prawn–rice system. Water levels were 0.3–0.6 m in rice growing areas. The water temperatures at noon in the hot months were higher than the suitable range for prawns. The final mean weights of prawns in treatments 1, 2 and 3 PL m^?2 were significantly higher than in treatment 4 PL m^?2 (P<0.05). The yields from different density treatments ranged from 194±82 to 373±32 kg ha^?1 and increased significantly with the increases in prawn densities (P<0.05). There was no significant difference between the two types of feed on production parameters (P>0.05). The total cost significantly increased at higher densities (P<0.05), but it was not different between feed types (P>0.05). Net profit and cost benefit ratio of treatment PS were significantly higher than in treatment P. In the present study, treatment 3 PL m^?2 and treatment PS offered the optimal results.