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.     

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.     

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.     

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.     

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.     

Yield characteristics and morphometric relationships of giant freshwater prawn,Macrobrachium rosenbergii (de Man, 1879)

Meat yield of 233 individuals of giant freshwater prawn, Macrobrachium rosenbergii (de Man, 1879), and their morphometric relationships were examined. Morphometric parameters of different length and weight measurements were recorded for both the sexes separately and subjected to analysis. The meat yield of prawns was size and sex dependent with females showing significantly greater (P < 0.05) tail meat yield than males. The tail meat as a body weight percentage ranges from 26.12 to 40.75 and 32.97 to 42.11 in males and females, respectively. Much of the differences in yield are related to the size and weight of the head and claws. The head weight of males was observed to be higher compared to females in all the size classes. Regression slopes of total length versus body weight and total length versus head weight were significantly higher than critical isometric value indicating high positive allometric growth in both the sexes, with males showing significantly higher (P < 0.05) slope than females. The relationship parameters of total length versus tail weight and total length versus tail meat showed a positive allometric growth in females, whereas it is isometry and negative allometry in males. The morphometric relationship of tail length versus all weight measurements showed positive allometry and negative allometry in females and males, respectively.     

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.     

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.     

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.     

A test of two methods for marking larvae and postlarvae of the giant freshwater prawn, Macrobrachium rosenbergii

This paper describes the successful use of two tagging systems, both produced by Northwest Marine Technology Inc., on larval and postlarval giant freshwater prawns, Macrobrachium rosenbergii. Visible implant (VI) elastomer tags (a coloured liquid that solidifies under the epidermis) were used on stage XI larval prawns (mean weight 0.01 g) and postlarval prawns (mean weight 0.07 g). VI alphanumeric tags (small biocompatible plastic labels also inserted under the epidermis) were tested on postlarval prawns (from a weight of 0.5 g). Tags were inserted using clove oil as anaesthetic, and survival, mortality and growth rates of tagged animals were compared with those of controls that were handled but not anaesthetized or marked. Twenty per cent of the larval prawns (the smallest of the group) died just after tagging, but thereafter the remaining prawns survived well, as did all the tagged postlarval prawns. Visibility of the VI elastomer tags in larval prawns deteriorated with time, though 79% of marks were still visible to the naked eye 70 days after tagging. VI elastomer tags in the postlarval group remained clearly visible for up to 100 days. Visibility of the VI alphanumeric tags fell shortly after tagging, but remained adequate thereafter. Moult rates in control and tagged animals were the same in larvae with VI elastomer tags and postlarvae with VI alphanumeric tags, but the moult rate in the postlarval prawns given elastomer tags was slower than in controls. Rates of growth were similar in tagged (elastomer and alphanumeric) and control postlarval prawns, once the size‐dependent mortality of tagged larval prawns was taken into account. We conclude that VI elastomer tags could be used to mark small numbers of individual larval and immediately postlarval prawns for periods of several months, and that VI alphanumeric tags could be used to mark an unlimited number of individuals from a size of approximately 0.5 g.     

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.     

Effect of feeding Lactobacillus-based probiotics on the gut microflora, growth and survival of postlarvae of Macrobrachium rosenbergii (de Man)

The present study was conducted to study probiotics treatment in the post‐larval diet of Macrobrachium rosenbergii. Three hundred postlarvae (average weight, 114–118 mg±0.11) were divided in five experimental groups each with four replicates. The experiment was conducted for 60 days. Experimental diets were identical in all the aspects except for variation in the probiotics strain. T1 and T2 groups were fed Lactobacillus acidophilus (140×10¹¹ CFU 100 g^?1) and L. sporogenes (24×10⁷ CFU 100 g^?1) respectively. The T3 group was fed L. sporogenes bioencapsulated in Artemia nauplii. T4 was the control group (without probiotic) whereas T5 was fed Artemia along with control diet. The bacteriological study indicated that the gut microflora of postlarvae are devoid of lactic acid bacteria. The probiotic strains were found to have inhibitory effects against the gram‐negative bacterial flora present in the gut. Growth of the probiotic fed groups was significantly higher (P<0.05) than the control group. Significantly higher growth (P<0.05), per cent weight gain (132.5%), specific growth rate (1.41%), feed efficiency ratio (FER) (0.45), protein efficiency ratio (1.29) and protein gain (161.6%) were recorded in T3 group fed Artemia bioencapsulated L. sporogenes over the control group (P<0.05). Although insignificant (P>0.05), growth‐promoting effects of L. sporogenes were found to be higher than L. acidophillus. Survival of the postlarvae was not affected by probiotics in the diet.     

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.     

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.     

Use of Cyclop-eeze as a substitute for Artemia nauplii in larval rearing of giant freshwater prawn Macrobrachium rosenbergii (De Man 1879)

Four feeding experiments, replacing 25% (T₁), 50% (T₂), 75% (T₃) and 100% (T₄), by dry weight, of the live feed Artemia nauplii for Cyclop‐eeze, a new larval feed that was claimed to contain the highest known levels of astaxanthin and omega‐3 polyunsaturated fatty acids, were compared against a control that was fed with Artemia and egg custard alone, to the larvae of giant freshwater prawn Macrobrachium rosenbergii (De Man 1879). Analysis of different production characteristics of the larvae revealed that the highest survival up to postlarvae (PL) stage was obtained for T₂ in which 50% of the Artemia nauplii were replaced by Cyclop‐eeze [freeze‐dried (FD) deep frozen (DF)], and the highest astaxanthin content of the larval tissue obtained in T₄ in which the larvae were fed 100% Cyclop‐eeze, although the survival rate was the lowest in this treatment. The costs of different treatments were also compared. The Artemia consumption million⁻¹ larvae was the highest in control (11490 g), followed by T₁ (8240 g), T₂ (4990 g), T₃ (3730 g) and T₄, which completely replaced Artemia from stage 5 onwards (1830 g). The highest consumption of Cyclop‐eeze million⁻¹ larvae was in T₄ (1670 and 10 880 g), followed by T₃ (850 and 5560 g), T₂ (410 and 2690 g) and T₁ (230 and 1490 g) of FD and DF, respectively. The astaxanthin contents of the late‐stage larvae fed under the four treatments were 24.90, 27.40, 28.60 and 35.60 μg g⁻¹ tissue for T₁, T₂, T₃ and T₄, respectively, while that of the control was 23.70 μg g⁻¹. The lowest cost of live feeds million⁻¹ PL was obtained for T₂ (US$ 428.60), followed by T₁ (US$ 490.46), control (US$ 529.07) and T₃ (US$ 583.26), while it was the highest for T₄ (US$ 890.93). The results indicated that Cyclop‐eeze could economically replace Artemia nauplii at 50% level that could significantly improve the survival and carotenoid composition of the larvae of M. rosenbergii.     

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]