Carbonic anhydrase and Na/K-ATPase activities at different molting stages of the giant freshwater prawn <Emphasis Type="Italic">Macrobrachium rosenbergii</Emphasis>

ABSTRACT:   In the present study, the role of carbonic anhydrase (CA) and Na/K-ATPase in the gill and epidermal tissues in the giant freshwater prawn Macrobrachium rosenbergii was examined as a function of molting stage. CA activity levels in the front and back gills were low at the intermolt stage C₀, but increased significantly at premolt stage D₃, and then decreased after molting. In the epidermal tissue, activity levels decreased gradually towards premolt to a minimum level at stage D₃, but became elevated at postmolt stages A and B. Na/K-ATPase levels in the front and back gills did not change significantly during the molt cycle. CA in the gill is possibly involved in supplying counter-ions for ion uptake, while CA in the epidermal tissue may play a role in mineralizing the exoskeleton after ecdysis. Na/K-ATPase in the gills may function in ion uptake from the ambient medium; however, since its activity was not influenced by the molt cycle, it probably does not have a major role in osmoregulation in the freshwater environment.     

Effect of various medium compositions on survival and hatching rates of embryos of the giant freshwater prawn Macrobrachium rosenbergii cultured in vitro

SUMMARY: The effectiveness of in vitro embryo culture of the giant freshwater prawn Macrobrachium rosenbergii depended on the age of the embryos at the onset of culture and on the concentrations of various compositions in the medium. Embryos that started being cultured on day 0.5 after oviposition were more sensitive to variations in the medium compositions than those that started being cultured on day 10.5 after oviposition. An optimal NaCl level was essential for embryonic development, survival, hatching and survival of the newly hatched larvae. Variations of NaCl or KCl levels dramatically altered embryonic development, and variation of the MgCl₂ + MgSO₄ level significantly lowered survival of the embryos that started being cultured at the early stage of development. In contrast, no significant change in embryonic development was observed upon variation of the CaCl₂ level. Hatching of the embryos required the presence of NaCl and CaCl₂ but not KCl or MgCl₂ + MgSO₄. The ionic requirements of the newly hatched larvae differed from that of the developing embryos. Variations of NaCl, KCl or CaCl₂ but not MgCl₂ + MgSO₄ levels significantly influenced the survival of the newly hatched larvae.     

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.     

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.     

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。     

罗氏沼虾胚胎附着机制的初步研究

甲壳动物十足目的大多数种类,其受精卵自产出后就附着于亲体腹部,并完成胚胎发育过程直至孵化,这对于胚胎的发育、保护和幼体散布有重要的生物学意义。胚胎的附着机制较为复杂,与刚毛、卵柄、卵索、卵膜和黏液腺有关:卵柄和卵索的形成决定了胚胎发育的顺利进行;卵膜的变化影响着胚胎的附着;黏液腺在胚胎附着过程中形成卵柄和外层卵膜,胚胎是依靠卵柄或卵索附着于腹肢的刚毛上,进而完成胚胎发育过程。本研究采用常规组织学和扫描电镜的方法研究了罗氏沼虾(Macrobrachium rosenbergii)胚胎及其附着结构。结果表明,胚胎附着与母体腹肢的结构、黏液腺分泌的黏液相关,也与胚胎表面结构、次级卵膜的形态和结构密切相关。原肢底节和内肢刚毛密而长,基部有细小的分支,有利于胚胎的附着,外肢刚毛羽状则有利于胚胎的托附和保护。抱卵雌虾腹肢的原肢和内肢上布满腹肢黏液腺分泌的黏液;携卵刚毛中空、壁厚,刚毛外壁由微绒毛状结构组成,扩大了携卵刚毛的表面积,以利胚胎附着。胚胎外表面由初级卵膜和次级卵膜组成,次级卵膜主要由卵巢黏液腺分泌的黏液参与形成。排卵时,受精卵依靠坚韧的次级卵膜相互粘连,同时,部分受精卵在刚毛上滑动、旋转,并借助次级卵膜逐渐形成了卵柄或卵索固着在腹肢刚毛上。从受精卵到胚胎孵化,胚胎之间最主要的连接方式是依靠次级卵膜的连接。胚胎的表面有许多褶皱和黏液,利于胚胎的附着以及胚胎对溶氧和小分子物质等的通透。以上结构与附着相适应,保证了胚胎发育的顺利进行和幼体的正常孵化。本研究旨为进一步认识甲壳动物胚胎附着机制、胚胎发育和人工育苗的研究提供参考。[中国水产科学,2007,14(1):67-73]     

Engulfed pathogen-induced apoptosis in haemocytes of giant freshwater prawn, Macrobrachium rosenbergii

Haemocytes of the giant freshwater prawn, Macrobrachium rosenbergii, were investigated for the induction of apoptosis after phagocytosis of pathogenic yeasts, bacteria and non-pathogenic latex beads in vitro. Isolated haemocytes of M. rosenbergii were cultured at a ratio of 1:50 haemocytes to pathogen with the yeast Debaryomyces hansenii, the bacteria Aeromonas hydrophila or Enterococcus faecium, or with latex beads at 25 degrees C for 2 h, followed by washing to remove free particles. At least 200 haemocytes were counted to determine the phagocytosis rate, and the results showed that haemocytes engulfed latex beads at a higher rate than the aquatic pathogens. By transmission electron microscopy, the yeast- or bacterium-engulfing haemocytes displayed morphological changes characteristic of apoptosis, including formation of cytoplasmic vacuoles, chromatin condensation and fragmentation of nuclei. This pathogen-induced apoptosis was further confirmed by DNA laddering and TUNEL (terminal deoxynucleotidyl transferase-mediated deoxy-UTP nick-end-labelling) assays. Neither haemocytes treated with latex beads nor uninfected haemocytes (control group) showed signs of apoptosis after 48 h in culture.     

生物絮团对罗氏沼虾体组成和消化酶活性的影响

通过向养殖水体中泼洒糖蜜构建生物絮团养殖模式,分析生物絮团营养组成,并探讨生物絮团对罗氏沼虾体组成和消化酶活性的影响。试验分对照组和试验组(生物絮团组),其中试验组在养殖过程中泼洒糖蜜。试验在室内水泥池内(2 m×2 m×0.6 m)进行,每个处理有3个重复,每个重复225尾虾(0.26 g±0.02 g),试验周期为90 d。养殖过程中不换水,糖蜜的泼洒量根据饲料投喂量进行计算(C/N为20)。结果显示:添加糖蜜能够显著促进生物絮团的形成,到第90天时,试验组的絮团体积达21.22 mL/L;而对照组为6.03 mL/L;试验组絮团粗蛋白含量为29.47%,粗脂肪含量为4.32%,二者均显著高于对照组,而粗灰分含量为11.36%,显著低于对照组;泼洒糖蜜对罗氏沼虾体组成的影响不显著,对照组和试验组肌肉粗蛋白含量分别为21.09%和21.20%,粗脂肪含量分别为2.91%和3.06%;另外,向水体中泼洒糖蜜对罗氏沼虾消化酶活性影响显著。试验组罗氏沼虾肠脂肪酶活性、胃脂肪酶活性和胰脂肪酶活性均显著高于对照组;试验组罗氏沼虾糜蛋白酶活性、胰蛋白酶活性也均显著高于对照组。但泼洒糖蜜对肠淀粉酶、胃蛋白酶、胃淀粉酶、胰淀粉酶和纤维素酶活性没有显著影响。试验表明,生物絮团营养组成丰富,能够有效提高消化酶活性。     

外源Fe3+诱导罗氏沼虾铁蛋白mRNA的表达

探讨铁蛋白ferritin基因在罗氏沼虾中的组织表达分布及外源Fe3+对ferritin mRNA表达的影响。通过RT-PCR表明,ferritin mRNA在罗氏沼虾精巢、卵巢、肌肉、心脏、中肠腺和血淋巴中均有表达;荧光定量PCR结果证实了ferritin mRNA在中肠腺中表达量最高,其次是精巢。对罗氏沼虾进行FeCl3溶液肌肉注射后,荧光定量PCR结果表明,肌肉中ferritin mRNA表达量在注射后3 h增加了61倍,高水平的表达一直维持到8 h,随后在12 h表达量急剧地下降;卵巢中ferritin mRNA表达量在3 h有近4倍显著增加,在注射后48 h内一直维持在较高水平;心脏ferritin mRNA表达量在24 h后增加了8倍,48 h后表达量开始下降;在血淋巴、中肠腺和精巢注射前后均未发现有显著性差异(P>0.05)。以上实验结果表明,外源Fe3+可以诱导罗氏沼虾铁蛋白mRNA的表达。     

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.     

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.     

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.     

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.     

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.