Improved techniques for rearing mud crab Scylla paramamosain (Estampador 1949) larvae

A series of rearing trials in small 1 L cones and large tanks of 30–100 L were carried out to develop optimal rearing techniques for mud crab (Scylla paramamosain) larvae. Using water exchange (discontinuous partial water renewal or continuous treatment through biofiltration) and micro‐algae (Chlorella or Chaetoceros) supplementation (daily supplementation at 0.1–0.2 million cells mL⁻¹ or maintenance at 1–2 millions cells mL⁻¹), six different types of rearing systems were tried. The combination of a green‐water batch system for early stages and a recirculating system with micro‐algae supplementation for later stages resulted in the best overall performance of the crab larvae. No clear effects of crab stocking density (50–200 larvae L⁻¹) and rotifer (30–60 rotifers mL⁻¹) and Artemia density (10–20 L⁻¹) were observed. A stocking density of 100–150 zoea 1 (Z1) L⁻¹, combined with rotifer of 30–45 mL⁻¹ for early stages and Artemia feeding at 10–15 nauplii mL⁻¹ for Z3–Z5 seemed to produce the best performance of S. paramamosain larvae. Optimal rations for crab larvae should, however, be adjusted depending on the species, larval stage, larval status, prey size, rearing system and techniques. A practical feeding schedule could be to increase live food density from 30 to 45 rotifers mL⁻¹ from Z1 to Z2 and increase the number of Artemia nauplii mL⁻¹ from 10 to 15 from Z3 to Z5. Bacterial disease remains one of the key factors underlying the high mortality in the zoea stages. Further research to develop safe prophylactic treatments is therefore warranted. Combined with proper live food enrichment techniques, application of these findings has sustained a survival rate from Z1 to crab 1–2 stages in large rearing tanks of 10–15% (maximum 30%).     

Development of nursery culture techniques for the mud crab Scylla paramamosain (Estampador)

Cannibalism is one of the main causes of mortality in the culture of the mud crab Scylla paramamosain, particularly in the early post‐larval and juvenile stages when the densities of hatchery‐reared crabs may be very high before stocking into ponds or release into the wild for stock enhancement. In a series of experiments investigating cannibalism mitigation, the influence of stocking density, the effectiveness of sand substrate, brick and shell shelters and feed type were compared in culture of crabs from instar 1 for short nursery periods of 15–30 days. Inclusion of brick and shell shelters significantly increased survival over sand substrate alone. However, inclusion of shelters did not affect growth rates. In scaled‐up nursery production in lined‐ponds, with shelters, live Artemia biomass and fresh chopped shrimp or tilapia were found to be equally effective feeds for juvenile crabs stocked at a density of 70 m⁻² from instar 1 and grown for 30 days [52–66% survival, 21.6–24.6 mm carapace width (CW)]. In an extended nursery period for a further 30 days, crabs of 22 mm CW, stocked at 30 m⁻² in the same ponds, attained a final size of 34.5–36.2 mm CW with a survival of 64.3–67.0% using the same feeds.     

Effects of different dietary lipid sources on tissue fatty acid composition,serum biochemical parameters and fatty acid synthase of juvenile mud crab Scylla paramamosain (Estampador 1949)

A 6‐week study was conducted to determine the effects of different lipid sources in pelleted diets on juvenile mud crab Scylla paramamosain. Five isonitrogenous and isolipidic diets containing 8% level of fish oil (FO), lard (LD), safflower oil (SO), perilla seed oil (PO) or mixture oil (MO; V_FO:V_SO:V_PO = 1:1:1), and a live food of marine bivalve Potamocorbula rubromuscula as the control diet (CF), were fed to groups of 25 juvenile crabs (average initial weight 7.4 g, carapace width 3.5 cm) in triplicate. The results showed that crabs fed MO had the highest survival (P < 0.05). The moisture content was significantly higher in crabs fed LD, SO and PO (P < 0.05). Crabs fed SO exhibited the lowest crude protein and lipid (P < 0.05). Ash contents were obviously lower in LD group (P < 0.05). Highest total lipid in the hepatopancreas and muscle was in LD and FO group respectively. Glucose, total cholesterol and low‐density lipoprotein were higher while high‐density lipoprotein was lower (P < 0.05) in LD group. Tissue fatty acid compositions were consistent with those in diets. FO and MO diets had the same depression effect like CF on fatty acid synthase activity and mRNA expression in the hepatopancreas. The results of this study indicated that FO and mixed oil are suitable for preparation of pelleted diets with better effects for juvenile S. paramamosain compared with live food, and the ratio of n‐6/n‐3 fatty acids in pelleted diets must be <1.     

Induced out-of-season spawning of the mud crab, Scylla paramamosain (Estampador) and effects of temperature on embryo development

Treated with combined bilateral eyestalk ablation and maintenance of water temperature at 22.5±1.5 °C, mud crab Scylla paramamosain females with mature ovaries were induced to produce eggs outside the natural spawning season in subtropical southern China. Newly extruded eggs from a crab were incubated in vitro at 10, 15, 20, 25, 27, 30, 35 °C, respectively, and the embryonic development was closely monitored. Abnormal cell division was observed at temperatures 10 and 35 °C. At 15 °C, development remained at the gastrula stage by day 32 post‐spawn, at which time the experiment was terminated. Hatching of in vitro incubated eggs occurred between 20 and 30 °C. An increase in incubation temperature from 20 to 25 °C reduced the incubation duration by 14 days, 2.6 times of that measured for a similar 5 °C increase from 25 to 30 °C. Embryonic development of S. paramamosain was divided into stage 0–10, and the duration of each stage was recorded for each incubation temperature. The information obtained allows accurate prediction of hatching time of female crabs incubated under variable temperatures. Larvae hatched from in vitro incubated eggs were reared to reach first juvenile crab stage and their dry weights were similar to those of larvae hatched naturally.     

Comparative performance of hatchery-reared and wild Scylla paramamosain (Estampador, 1949) in pond culture

The performance of hatchery‐reared juveniles either in aquaculture grow‐out systems or stock enhancement is likely to be dependent on a range of factors during the hatchery phase of production. With recent progress in the development of hatchery systems for the mud crabs Scylla spp., there is growing interest in evaluation of the quality of hatchery‐reared juveniles relative to wild seed crabs as currently used in aquaculture. Hatchery‐reared and wild‐collected Scylla paramamosain juveniles were stocked either together in ponds or separately. All crabs were tagged with microwire‐coded tags, so that origin could be determined in the mixed groups. Preliminary validation demonstrated that tagging did not affect survival or growth, with a tag retention of 94%. After 106 days of culture, there was no significant difference in survival at harvest between the two sources of crabs. The wild juvenile crabs had a significantly higher initial weight:carapace width (CW) ratio compared with those from the hatchery, indicating a difference in condition. However, where crabs were stocked separately, the hatchery‐reared animals exhibited significantly faster growth than those collected from the wild, both in terms of specific growth rate and CW increase per month. However, in the mixed ponds, where there was competition with wild crabs, there was no significant difference in growth rate between crabs from the two sources. Overall, the results demonstrate that the growth performance of hatchery‐reared S. paramamosain can at least equal that of wild‐collected seed crabs in ponds culture.     

Parentage determination of the mud crab Scylla paramamosain using microsatellite markers

This study evaluated the properties of nine Scylla paramamosain microsatellite loci screened by us previously for inclusion in a parentage assignment marker suite. These nine highly polymorphic markers (mean He=0.847 and PIC=0.830) were determined as being suitable for parentage assignment. Simulations based on allele frequency data from 15 known maternal families (165 individuals) demonstrated that at least four loci were required to assign >95% of offspring to maternal parents with 95% confidence. In actual parentage assignments, all progenies were assigned to the maternal parents with six or more loci, which was similar to the simulation predictions. Our results suggest that this set of microsatellites provide a powerful and efficient tool for identifying pedigree information for selective breeding programmes of S. paramamosain.     

利用微卫星标记比较分析拟穴青蟹不同家系的遗传多样性

拟穴青蟹(Scylla paramamosain)是我国重要的海水养殖蟹类,随着市场需求量的增加及养殖规模的不断扩大,有必要培育高产、抗逆的优质新品种.家系选育是选择育种的有效途径,目前已应用于拟穴青蟹人工育种工作中.本研究利用8个微卫星标记对拟穴青蟹6个家系共275 ind个体进行了遗传多样性分析,共检测到50个等位...     

Reproductive performance and offspring quality in mud crab (Scylla paramamosain) broodstock fed different diets

A 2-month feeding trial was conducted toevaluate the reproductive performance andoffspring quality of mud crab (Scyllaparamamosain) females fed either a mixture offresh food items (squid, shrimp, trash fish andArtemia biomass) or two experimentaldiets developed for penaeids. Before testinitiation, mud crab females with an averageindividual wet weight of 200–300 g wereacclimated for 2–3 days and reared together inone concrete tank of 2.0 × 0.5 × 8 m untilspawning. After spawning, the spent spawnerswere unilaterally eyestalk ablated and randomlydivided (20 animals/treatment) over three tanksof the same size and subjected to the dietarytreatments. Spent spawners were used toeliminate the effect of feeding history.There were only minor differences inreproductive performance between dietarytreatments. No differences were observed in theduration of the latency period from eyestalkablation to spawning. Fecundity was onlymarginally higher for the broodstock fed thecontrol diet. Also egg quality seemed onlyslightly affected by the treatments. Egghatching rates were slightly higher in crabsfed the formulated diets compared to thosecrabs fed the fresh diet. The onlystatistically significant difference (p < 0.05) observed however was in egg hatchingrate between the control diet and diet A2. Incontrast, the crabs fed the fresh diet producedstronger larvae as determined by a starvationtest.We therefore conclude that artificial dietsresulted in reproduction success comparable tothe use of fresh food. The nutritionalcomposition of the artificial diets couldhowever be improved in order to produce larvaeof optimal quality. Based on our researchfindings, the protein level and n-3 HUFA levelin the diet warrants further investigation inthis respect.     

Effects of dietary phospholipids levels on growth performance,lipid metabolism,and antioxidant capacity of the early juvenile green mud crab,Scylla paramamosain (Estampador)

Four iso‐nitrogenous and iso‐lipidic diets were designed to investigate the effects of dietary phospholipids (PL) levels (with 0%, 1%, 2%, and 4% PL supplementation) on growth performance, lipid metabolism, and antioxidant capacity in early juvenile green mud crab (Scylla paramamosain). There were three replicates of 28 crabs (initial body weight from 42.02 to 42.44 mg) for each diet treatment, and growth trial lasted for 8 weeks. At the end of the growth trial, there was no significant difference in survival among all treatments. Crabs fed diet with 2% PL obtained highest weight gain (WG) and specific growth rate than other crabs. The molting frequency was not affected by different dietary PL addition. Besides, the contents of whole body lipid and long chain highly unsaturated fatty acids significantly increased with elevating dietary PL levels. In the hepatopancreas, crabs fed diet with 0% PL had significantly higher malondialdehyde concentration than other crabs. And crabs fed diet with 2% PL obtained significantly higher superoxide dismutase activity than crabs fed diets with 0% and 4% PL. Moreover, the mRNA expression of Na⁺/K⁺‐ATPase was significantly down‐regulated with dietary PL supplementation over 2%. Based on the second order polynomial regression analysis of WG, 2.37% dietary PL level was the optimal demand for early juvenile S. paramamosain. Moreover, we found crabs fed diet with 2% PL obtained better antioxidant capacity than other crabs.     

拟穴青蟹vasa基因cDNA的特征分析与性腺特异表达

vasa蛋白属于DEAD家族蛋白,是一种RNA解旋酶,在生殖细胞的分化中具有重要作用。本研究从拟穴青蟹卵巢中克隆得到vasa基因cDNA全长(将其命名为Sp-vasa),Sp-vasa序列长度为2 944 bp,其中开放阅读框(ORF)长度为1 899 bp,编码632个氨基酸,具有DEAD-box家族蛋白的7个高度保守的结构域。半定量RT-PCR结果显示,Sp-vasa只在卵巢和精巢中特异表达,在其它组织则未检测到表达。由此可见,vasa属于拟穴青蟹性腺特异表达基因。因此,Sp-vasa基因可有望作为有效的分子指标研究拟穴青蟹原始生殖细胞的起源、迁移以及生殖腺的发生。     

Overwintering behaviour affects distinctive flavour of mud crab Scylla paramamosain in commercial farms

In order to reveal the influence of overwintering behaviour on the flavour of Scylla paramamosain, this study selected crabs of before and after overwintering in SM (Sanmen County, Zhejiang Province), HP (Hepu County, Guangxi Province) and WN (Wanning city, Hainan Province). The contents of flavouring‐related substances in hepatopancreas, muscles and gonads were determined by high‐performance liquid chromatography (HPLC); taste activity value (TAV) and equivalent umami concentration (EUC) were calculated. The results showed that sweetish amino acids were mainly accumulated in muscles, umami amino acids were mainly accumulated in hepatopancreas, free nucleotides were mainly accumulated in gonads, and the EUC value in gonads was the highest. After overwintering, the content of sweetish, umami amino acids and free nucleotides in muscle and hepatopancreas decreased significantly, while the content of flavour‐tasting amino acids in gonads did not change significantly, but the content of umami nucleotides increased significantly. Therefore, overwintering behaviour decreased the flavour of muscle and hepatopancreas, slightly enhanced gonad and the overall flavour of S. paramamosain before overwintering was better than that after overwintering.     

Effect of different forms of Artemia biomass as a food source on survival,molting and growth rate of mud crab (Scylla paramamosain)

An experiment was conducted to evaluate the effect of different forms of Artemia biomass as a food source on survival, molting and growth rate of mud crab Scylla paramamosain. Instar 1 crablets with a mean weight of 0.0082 ± 0.0007 g were reared both individually and communally and fed with different diets consisting of fresh shrimp meat (control feed), live Artemia biomass, frozen Artemia biomass and a dried Artemia‐based formulated feed for 40 days. The highest survival was obtained for crablets receiving live Artemia (92.5% and 75.8%) followed by the groups fed with frozen biomass (90.0% and 47.5%), the control feed (72.5% and 24.2%) and the dried Artemia‐based diet (60.0% and 21.7%) for individual and communal cultures, respectively. The intermolt period, the total number of moltings and the growth rate, which were determined on individually reared crabs, showed the same pattern as for survival. The results suggest that crab performance decreased in the following order: live Artemia>frozen Artemia > fresh shrimp meat > dried Artemia‐based formulated feed. Live Artemia biomass proved an ideal feed for nursery of Scylla paramamosain crabs. Frozen Artemia biomass may be an alternative in times of shortage. Our findings illustrate the high potential for local utilization of Artemia biomass in Vietnam for reliable production of mud crab juveniles.     

锯缘青蟹苗种培育的科技进展

锯缘青蟹是我国南方沿海各省传统养殖的海水蟹类,在当前农业产业结构调整中被列为主要发展品种之一。鉴于苗种是限制青蟹养殖业发展的主要因素,本文根据目前的研究现状对青蟹的种质、性成熟机制,产卵、孵化条件,幼体培育过程中营养条件、盐度、温度、光照、疾病等,以及从大眼幼体到仔蟹的中间培育技术等作了简要归纳,并对研究现状和进入产业化急需解决的问题进行了探讨。     

大黄多糖对拟穴青蟹免疫相关基因表达的影响

运用RT-PCR技术探索拟穴青蟹(Scylla paramamosain)注射免疫增强剂——大黄多糖(rhubarb polysaccharide,RP)后11个免疫相关基因的表达变化情况。结果发现,大黄多糖刺激拟穴青蟹4 d后,血细胞中4个基因(丝氨酸蛋白酶抑制因子、过氧化氢酶、过氧化物还原酶和酚氧化酶原)表达水平分别上调1.1倍、4.1倍、1.3倍和2.1倍,与对照组相比,存在极显著(P0.01)或显著性差异(P0.05)。肝胰腺中5个基因表达变化显著(P0.05),其中丝氨酸蛋白酶抑制因子、过氧化物还原酶和抗脂多糖因子3个基因表达分别上调2.1倍、4.8倍和1.7倍;过氧化氢酶和溶菌酶仅在实验组中表达,未见其在对照组表达。由此推测,丝氨酸蛋白酶抑制因子、过氧化氢酶、过氧化物还原酶、酚氧化酶原、抗脂多糖因子和溶菌酶6个基因可能与拟穴青蟹免疫增强有关。     

Pond production of the mud crabs Scylla paramamosain (Estampador) and S. olivacea (Herbst) in the Mekong Delta, Vietnam, using two different supplementary diets

The production performance of Scylla paramamosain and S. olivacea reared together in ponds and provided with different supplementary diets (crustaceans or trash fish) were evaluated with that of an unfed control group relying only on natural food available within the pond. Each treatment was conducted in triplicate. The aim was to evaluate two production strategies over a 130‐day grow‐out trial: (1) production to 200 g crab^?1, which is the minimum preferred marketable size in the Lower Mekong Delta (LMD) and (2) production to 300 g crab^?1, the size that fetches the highest market price. Growth production models were estimated which revealed that the rearing period required to produce crabs of 200 g was 102 days with supplementary feeding, compared with 120 days for the unfed control group. The rearing period needed to produce 300‐g‐sized crabs was estimated to be 144 days and 186 days for the supplementary fed and unfed groups respectively. The additional risk involved in rearing mud crabs to 300 g is not considered worthwhile, under similar culture conditions as observed in this trial, as a decrease in pond water quality and higher mortality rates were observed in the final month of culture needed to achieve this larger size.     

Lipids and fatty acids in wild and pond-reared mud crab Scylla serrata (Forsskål) during ovarian maturation and spawning

Wild‐caught and pond‐reared female mud crab Scylla serrata at different stages of ovarian maturation were collected from Samar and Capiz, Philippines. Crabs were categorized into five stages according to the external morphological and microscopic appearance of the most advanced oocytes. The ovaries, hepatopancreas, muscle and newly spawned eggs (NSE) were analysed for lipid class components and fatty acids. Total lipid was higher in pond‐reared than in wild‐caught crabs but increased with ovarian maturation in both groups. Ovarian lipid peaked at the fully mature stage, coinciding with a decline in hepatopancreatic and muscle lipids. Lipid levels declined significantly in spent females. The tissues contained elevated highly unsaturated fatty acids such as arachidonic (20:4n‐6), eicosapentaenoic (20:5n‐3) and docosahexaenoic (22:6n‐3) acids, but at higher levels in late maturing and fully mature ovaries and in NSE. The type of lipid class and fatty acid components in mature ovaries as well as in NSE are generally considered to be indicative of their importance in reproductive physiology and embryonic and larval development.     

Reproductive performance, lipids and fatty acids of mud crab Scylla serrata (Forsskål) fed dietary lipid levels

Natural food (NF, control), artificial diets (AD) containing total lipid levels of 10%, 12% and 14% (AD10, AD12 and AD14) and their combinations (AD10+NF, AD12+NF and AD14+NF) were fed for 112 days to pond‐sourced eyestalk‐ablated mud crab Scylla serrata (625±6.4 g) in tanks in order to determine their effects on reproduction and lipid profiles in broodstock tissues and zoeae. Crabs fed NF had the highest number of spawning followed by crabs fed AD10+NF and AD14+NF. Higher offspring production (number of zoeae) was obtained from crabs fed NF and AD+NF than from AD. As dietary total lipid levels increased, total lipid of broodstock ovaries, hepatopancreas, muscle and zoeae correspondingly increased in which AD+NF promoted higher levels than AD. Increased dietary total lipid levels enhanced lipid classes such as triacylglycerols and phosphatidyl choline levels in zoeae, all higher in crabs fed AD+NF than in AD. The major fatty acids in zoeae, particularly 16:0, 18:0, 18:1n‐9 and 20:4n‐6, 20:5n‐3 and 22:6n‐3, were higher in crabs fed AD+NF than in AD, the contents corresponding to broodstock dietary total lipid levels. A 10% total lipid in AD in combination with NF was sufficient to provide the essential lipids in crabs in the improvement of larval production and quality.