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.     

Growth and carcass composition of giant freshwater prawn, Macrobrachium rosenbergii (De Man), fed different isonitrogenous and isocaloric diets

A feeding experiment was conducted for 135 days to observe the effect of different isonitrogenous (35% crude protein) and isocaloric (385 kcal) diets on the growth and carcass composition of giant freshwater prawn, Macrobrachium rosenbergii ( De Man 1879 ). Three experimental diets (ED1, ED2 and ED3) were prepared using locally available ingredients. These diets differed mainly in terms of percent contribution of major protein sources such as fish meal, soybean meal, groundnut oil cake and mustard oil cake. Experimental diets were evaluated against a commercial diet, which served as the control (CD). Juveniles 1.87–2.44 g in size were stocked at a population density of 40 000 ha⁻¹ and fed thrice daily at 10% in the beginning and reducing gradually to 7% and 5% of the body weight during the experimental period. No significant differences (P>0.05) in the growth performance were observed; however, a significantly (P<0.05) higher yield (721.9 kg ha⁻¹ 135 days⁻¹) was recorded for prawn fed with control diet, followed by experimental diet 2 (676.5 kg ha⁻¹ 135 days⁻¹, having soybean meal as a major protein source). The survival ranged between 63.8% and 77.7%, with different diets showing significantly higher survival. The apparent feed conversion ratio values of diets ranged between 3.15 and 3.49, with experimental and control diets showing non‐significantly lower AFCR values. At the end of the experiment, representative specimens from each treatment were collected and their carcass composition was measured. Analysis of variance showed that carcass protein and total carbohydrate contents were significantly (P<0.05) higher in prawns fed on a fish–soybean meal‐based diet (ED3) and a control diet. The total lipid contents of prawns, however, did not differ significantly among the various dietary treatments. The results of our study suggest that the experimental diets could be used effectively for M. rosenbergii without compromising growth and flesh quality.     

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.     

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.     

Economic analysis of monosex culture of giant freshwater prawn (Macrobrachium rosenbergii De Man): a case study

All‐male monosex culture of Macrobrachium rosenbergii (De Man) has emerged as a popular practice in India, especially in the state of Andhra Pradesh. A study was conducted to compare the economics of all‐male, mixed and all‐female culture in 15 adjacent, rectangular ponds of 4000 m² each by stocking juveniles previously reared in a nursery for 60 days. The experiment was conducted using a completely randomized design with three treatments; T₁ (all male), T₂ (mixed) and T₃ (all female), and five replicates for a period of 5 months after the nursery phase. Statistical analysis showed highly significant (P<0.01) differences among the three types of culture. The cost of production was estimated and the economic feasibility of the culture methods was evaluated by cost‐return and partial budgeting analysis. The average weight, productivity and specific growth rate were the highest for all male culture, being 80.92±2.41 g, 1532 kg ha^?1 and 1.97±0.02 respectively. All‐female culture registered significantly higher survival (89.16±0.77%) and the best apparent feed conversion ratio of 1.26±0.02. The economic analysis revealed that all‐male monosex culture of M. rosenbergii was 63.13% and 60.20% more profitable than mixed and all‐female cultures respectively.     

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

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

Osmoregulation,growth and moulting cycles of the giant freshwater prawn (Macrobrachium rosenbergii) at different salinities

The giant freshwater prawn, Macrobrachium rosenbergii, is a species with a high commercial value in aquaculture. Two experiments were performed to determine the effects of salinities on the osmoregulation, growth and molting cycles of M. rosenbergii during growout. The first experiment was designed to determine whether these animals are capable of adapting to the changes in salinity seen in salinity intrusions in tropical deltas, with an incremental increase in salinity of 3‰ per day from 0‰ to 30‰ Haemolymph osmolality was rapidly regulated up to salinities of 15‰ , whereas animals conformed at higher salinities. The second experiment determined the growth, moulting cycle, osmolality, muscle water content and mortality during a 4‐month experiment at 0‰, 15‰ or 25‰ salinity. The weight gains in 0‰ and 15‰ were not significantly different and were comparable to the growth rates achieved in production farms with body mass increases of 2.6 and 2.3‐fold their initial body mass, respectively, after 4 months. The 25‰ group suffered from low growth, high mortality and a significantly lower moulting frequency. These data show that this species can be reared in brackish water up to 15‰, allowing for farming in the large areas impacted by salt water intrusions in tropical deltas.     

Experimental susceptibility of different life-stages of the giant freshwater prawn, Macrobrachium rosenbergii (de Man), to white spot syndrome virus (WSSV)

Studies were conducted by injecting/feeding white spot syndrome virus (WSSV) derived from infected shrimp, Penaeus monodon (Fabricius), to different life-stages, namely post-larvae, juveniles, sub-adults and adults of Macrobrachium rosenbergii (de Man). The disease was also induced in brood stock, and the eggs and larvae derived from these animals were subsequently tested for WSSV infection. All the stages except egg used for the experiment were found WSSV positive in histopathology, cross infection bioassay and polymerase chain reaction (PCR) analysis. Experimentally infected post-larvae and juveniles showed a high percentage of mortality and an increased rate of cannibalism. The cumulative mortality in post-larvae was up to 28%; with 28–40% cannibalism resulting in a maximum loss of up to 68%. In juveniles, observed mortality and cannibalism were 10–20% and 6.7–30.0%, respectively, and the maximum loss recorded was 50%. In sub-adults, mortality ranged from 2.8 to 6.7%, cannibalism was up to 20% and the total loss was up to 26.7%. Sub-adults and adults were found to be more tolerant to the infection as evidenced by the mortality pattern. A nested (two-step) PCR resulted in a 570-bp product specific to WSSV in all stages, except the eggs.     

River pen culture of giant freshwater prawn Macrobrachium rosenbergii (De Man) in southern Vietnam

A technical and socio‐economic survey conducted in the Dong Thap province of Vietnam to assess the current status of river pen culture of giant freshwater prawn Macrobrachium rosenbergii (De Man) showed that pen culture, which has been developed and operated by farmers' indigenous knowledge, requires more study in order to optimize stocking density and to assess its environmental impacts. In this study, the prawn pens were of rectangular shape, with an average size of 209 m². Prawns were stocked in June at an average density of 62 pieces m^?2 and fed on farm‐made feed and were harvested 2–4 times starting from 4 months after stocking until late December or the following January. The average prawn yield was 0.52 kg m^?2 year^?1, ranging from 0.14 to 1.6 kg m^?2 year^?1. The average net return was US$0.71 m^?2 year^?1, ranging from US$1.24 to 4.37 m^?2 year^?1. About 73% of the farmers achieved positive net returns. The top five constraints for prawn pen culture were lack of knowledge of culture technologies, poor water quality, disease problems, poor quality of wild seed and lack of capital. Almost all farmers have no environmental awareness, and environmental regulations for prawn pen culture have not been established. Field measurements showed that all measured water quality parameters were within the range for good growth of giant freshwater prawns, and indicated that there was no significant accumulation of nutrients and organic matter at the bottoms of rivers or canals.     

Immunomodulatory effect of Cynodon dactylon against white tail disease of giant freshwater prawn,Macrobrachium rosenbergii (de Man, 1879)

The giant freshwater prawn, Macrobrachium rosenbergii, is an economically important and extensively cultured crustacean worldwide. The viral pathogens, Macrobrachium rosenbergii nodavirus (MrNV) and extra small virus (XSV) are responsible for causing severe mortalities in the hatchery and nursery phases. This study investigates the protection of postlarvae of freshwater against white tail disease (WTD) using plant extract derived from Cyanodon dactylon and the modulation of the prawn non‐specific immunity. To determine the immunomodulatory effect of C. dactylon extract, the prawn was injected with plant extract and various immunological parameters were estimated. The immunological parameters such as proPO, SOD, THC and clotting time were found to be significantly higher in the plant extract‐injected prawn when compared with control groups. The results of real time PCR analysis revealed up regulation on the expression proPO, SOD and lysozyme genes in MrNV and XSV challenged prawn postlarvae treated with C. dactylon extract. Infectivity experiment showed high relative per cent survival in MrNV and XSV‐challenged prawn postlarvae treated with C. dactylon extract. These results strongly indicate that the administration of C. dactylon plant extract enhances immunity of the prawn. Based on the results, this study recommends that the immersion of postlarvae in C. dactylon plant extract is a potential prophylactic agent against WTD.     

Effects of salinity on standard metabolic rate and critical oxygen tension in the giant freshwater prawn (Macrobrachium rosenbergii)

The extensively farmed giant freshwater shrimp, Macrobrachium rosenbergii, can survive salinities up to 26 g L^?1, but the commercially important grow‐out occurs exclusively in freshwater areas. Recent studies suggest the shrimp equally capable of growing in brackish as fresh water and a better understanding of how this species responds to changing salinity could significantly impact freshwater prawn farming in deltas and coastal areas. Here, the effect of salinity (0 and 15 g L^?1) on standard metabolic rate (SMR) and critical oxygen tension (P_crit) was measured in adult M. rosenbergii using intermittent closed respirometry. SMR was 79.8 ± 3.1 and 72.7 ± 2.9 μmol kg^?1 min^?1 in fresh and brackish water, respectively, with no significant difference between the two salinities (P = 0.122). During hypoxia M. rosenbergii maintained oxygen uptake down to a P_crit of 26.3 ± 1.4 mmHg in fresh and 27.2 ± 2.0 mmHg in brackish water (P = 0.682), showing that salinity had no overall effect on oxygen conductance in the animals. These findings are in agreement with recent growth studies and provide further evidence that grow‐out phase could be accomplished in brackish water areas. Thus, the predicted intrusions of brackish water in tropical deltas as a consequence of future global warming may not impact this important production.     

Liver oil of pharaoh cuttlefish Sepia pharaonis Ehrenberg, 1831 as a lipid source in the feed of giant freshwater prawn, Macrobrachium rosenbergii (De Man 1879)

The efficacy of pharaoh cuttlefish ( Sepia pharaonis Ehrenberg, 1831) liver lipid in the feed for juveniles of giant freshwater prawn Macrobrachium rosenbergii (De Man 1879) was tested by feeding five experimental diets prepared using clam meat, groundnut oil cake, wheat bran, tapioca powder, vitamin–mineral mixture and cellulose powder as the major ingredients. The test diets T₁, T₂, T₃, T₄ and T₅, containing five levels (1, 2, 3, 4 and 5%, respectively) of lipid, extracted from the liver of pharaoh cuttlefish, were fed to 30-day-old prawn juveniles for 60 days, in triplicates and compared against a control. Analysis of variance of the growth parameters showed that the juveniles fed with T₂, containing 2% cuttlefish liver lipid with a total lipid content of 9.85% and lipid–carbohydrate ratio of 1 : 3.8, showed significantly better growth ( P  < 0.05) with the highest weight gain of 118%, greatest food conversion and protein utilization efficiency, and the highest body protein content, although the survival rates remained unaffected among treatments. Growth of the juveniles, however, declined at lipid levels >2%. Addition of cuttlefish liver lipid led to an increase of ω-3 and ω-6 fatty acids in T₂, the proportion of ω-6 being higher than the former. The crude lipid content of the body of test animals also increased with the increase in dietary lipid from 1.51% in T₁ to 3.27% in T₅. The present study indicates that cuttlefish liver lipid can be successfully used as a substitute for conventional lipid sources in rearing juveniles of giant freshwater prawn, an eco-friendly practice in recycling cuttlefish waste – a discarded fishery by-product.     

人工诱导罗氏沼虾同步产卵与卵巢组织学研究

运用保幼激素类似物－ＺＲ５１５诱导罗氏沼虾同步产卵进行了生产性规模试验，每性雌虾体钙点滴４０μｌＪＨＡ－ＺＲ５１５，通过体表渗稼促进卵巢成熟和产卵。药物处理后１６－１７天，产卵率为４１．８１％，较常规生产的自然产卵率提高５０．２３％。罗氏沼虾卵巢生物学和组织滂研究表明，性成熟后的任何发育阶段，卵巢中均存在由卵原细胞组成的生发带和卵黄发生卵母细胞。     

Overview of recent research and development in temperate culture of the freshwater prawn (Macrobrachium rosenbergii De Man) in the South Central United States

Research and development efforts concerning freshwater prawn farming have in the past been principally conducted in tropical regions. However, activities in temperate regions also date back several decades. Culture of Macrobrachium in temperate zones offers positive opportunities, despite the inability to culture year‐round. Unique problems imposed by a restricted growing season must be addressed, and opportunities capitalized on, to attain commercially viable production in these regions. Much recent research in the South and Central USA has been directed towards intensifying production (kg ha^?1) without decreasing average harvest sizes or deteriorating water quality. Particular areas of research include evaluating and maximizing the relative contributions of natural foods, effects of artificial substrates on growth and prawn population structure and grading of animals before pond stocking to reduce heterogeneous individual growth and aggressive interactions. Recent studies have concentrated on combining these factors into a ‘best management practices’ (BMPs) production model. With these factors combined, production has increased from an average of 1000 kg ha^?1 of 30 g animals to almost 3000 kg ha^?1 of 40 g animals produced in research ponds in 110 days of culture. In commercial ponds, 1500–2000 kg ha^?1 has been achieved in a similar period using BMPs.     

Effect of cold shock and hormone on growth and male morphotypes of freshwater prawn,Macrobrachium rosenbergii (de Man)

A study was carried out to determine the effects of cold shock and 17α‐methyl testosterone hormone on growth in male morphotypes of giant freshwater prawn Macrobrachium rosenbergii. Juvenile males were exposed at 18 ± 0.25°C for 24 h with normal feed and cultured at 27 ± 0.7°C for 70 days as well as administrated feed with hormone separately at 400 mg kg^?1 feed for the first 30 days of culture period (70 days). The growth (Mean ± SD) of male juveniles at harvest was closely homogenous in the cold shock (CLS) treated group; while it was heterogeneous in hormone‐treated (MH) and control (CO) groups. Although total body weight of blue claw (BC) males was not significantly different in the controls and the treatments, the 2nd pereiopods (claw) weight and length of the BC was significantly lower in CLS (0.52 ± 0.05 g, 10.02 ± 0.65 cm) than that in the CO (0.71 ± 0.16 g, 12.53 ± 0.57 cm) and MH (0.75 ± 0.08 g, 12.85 ± 1.18 cm) treatments. Survival of prawns was similar in treatments. The morphotypes of male prawn were significantly different with 59% of BC males with small‐sized claws in CLS and, 45% and 30% of BC males with large‐sized claws in MH and CO respectively. Cold shock adversely affected male morphotypes of the prawn so that numbers of BC males with small‐sized claw was the highest in CLS tank. However, the CLS treatment resulted in more uniform marketable prawns and suggested a high production of BC males in an all‐male culture system.