Dietary protein requirement of giant mud crab Scylla serrata juveniles fed iso-energetic formulated diets having graded protein levels

The protein requirement of juvenile mud crab Scylla serrata (body weight=0.25±0.051 g, carapace width=9.3±0.04 mm) fed with different iso-energetic, iso-lipidic diets with graded protein levels (15–55% crude protein at 5% intervals) was determined. The feeding trial was conducted for a period of 63 days to determine the minimum and optimum protein requirement of juvenile S. serrata . The crabs fed with 15% and 20% dietary protein levels showed 100% and 12.5% of mortalities respectively. The mortalities observed in the above treatments were associated with the prolonged intermoult duration (46 and 32 days respectively). All other treatments recorded 100% survival. The best growth performance as well as the nutrient turn-over was recorded in crabs fed with 45% crude protein in the diet. Second-order polynomial regression of specific growth rate (SGR) as well as body protein gain vs. dietary protein levels suggested that 46.9–47.03% dietary protein is required for the best growth response and protein deposition in juvenile S. serrata . An extrapolation of 'SGR' and 'daily protein gain' upon the 'dietary protein level' axis ( Y =0) showed that 14.7–16.2% dietary protein is necessary for the minimum maintenance metabolism.     

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.     

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.     

Ovarian maturation stages of the mud crab Scylla serrata

Ovarian maturation in adult wild‐sourced and pond‐grown Scylla serrata (Forsskål) was determined based on gross morphology and histological appearance. There were no significant differences noted in the histological features of both wild and pond‐reared S. serrata females. Ovarian maturation was classified into five stages: immature, early maturing, late maturing, fully mature and spent. The immature ovaries are thin and translucent to off white and contain oogonia, primary oocytes with large nuclei. The follicle cells were found around the periphery of the lobes and an area among groups of oogonia and oocytes. The follicle cells gradually enclosed the oocytes. The early‐maturing ovaries were yellow and small yolk globules started to appear in larger oocytes. In late‐maturing ovaries, the colour became light orange and lobules were apparent. Yolk globules occurred in the cytoplasm with larger globular inclusions towards the periphery, while follicle cells were hardly recognizable. Fully mature ovaries were orange to deep orange and had swollen lobules. Large yolk globules were apparent in the entire cytoplasm. Follicle cells were hardly seen. Spent ovaries were similar to the early‐maturing and late‐maturing stage in partially spawned females. The ovarian development was correlated closely to the gonadosomatic index, oocyte diameter, and ovarian histology. The classification of ovarian maturation provides baseline information for further studies on reproductive biology. Likewise, the information provides a guide for broodstock management in the hatchery.     

Influence of binder type on leaching rate and ingestion of microbound diets by mud crab, Scylla serrata (Forsskål), larvae

Microbound diets (MBD), composed of nutrients held within a matrix or binder, were shown to be readily ingested by larvae of the mud crab, Scylla serrata (Forsskål). In an effort to improve experimental MBD developed for S. serrata larvae, this study determined the effects of various binders on estimated larval ingestion of MBD as well as their leaching rates. Microbound diets with the same dietary compositions were bound with either agar, alginate, carrageenan, gelatin or zein. All diets contained ¹⁴C‐labelled rotifers and were fed to Zoea I, III, V larvae and megalopae. The ¹⁴C content of larvae fed ¹⁴C‐labelled MBD for 2 h was used to estimate the relative ingestion rates of MBD. For all larval stages tested, the estimated mean larval ingestion of MBD did not differ significantly between binder types (P>0.05). Determination of the amount of ¹⁴C‐labelled nutrient leaching from MBD bound with various binders after 30, 60 and 240 min of immersion showed that for all types of binders, ¹⁴C leaching occurred primarily within the first 30 min of immersion. Zein‐bound MBD consistently showed the lowest numerical leaching rate among the five binders tested for all immersion periods examined, and the differences compared with other binders were often significant, particularly after 60 and 240 min of immersion. Larval feeding experiments showed that MBD prepared with binders showing higher rates of leaching were not utilized to a greater degree by S. serrata larvae. Furthermore, unnecessary leaching from MBD particles wastes dietary nutrients and is likely to impact on water quality. On this basis, our results suggest that because of its consistently lower leaching rates, zein is probably a more suitable binder for MBD developed for S. serrata larvae.     

Characterization of Aquimarina hainanensis isolated from diseased mud crab Scylla serrata larvae in a hatchery

Mass mortality due to necrosis signs occurred in hatchery-reared zoea stage larvae of the mud crab Scylla serrata in Okinawa, Japan, and a causative bacterium was isolated. In this study, we identified and characterized the bacterium by genome analysis, biochemical properties and pathogenicity. The bacterium was a Gram-negative, non-motile, long rod, forming yellow colonies on a marine agar plate. It grew at 20–33°C (not at 37°C) and degraded chitin and gelatin. Phylogenetic analysis of the 16S rRNA gene sequence identified the bacterium as Aquimarina hainanensis. Genome sequence data obtained from Illumina MiSeq generated 29 contigs with 3.56 Mbp in total length and a G + C content of 32.5%. The predicted 16 chitinase genes, as putative virulence factors, had certain homologies with those of genus Aquimarina. Experimental infection with the bacterium conducted on larvae of four crustacean species, brine shrimp Artemia franciscana, freshwater shrimp Caridina multidentata, swimming crab Portunus trituberculatus and mud crab S. serrata, revealed that this bacterium was highly virulent to these species. The present study suggests that the bacterium caused mass mortality in mud crab seed production was A. hainanensis and can be widely pathogenic to crustaceans.     

青蟹的营养需求研究及其配合饲料研制

青蟹属（Scylla）有锯缘青蟹（Scylla serrata），拟穴青蟹（S．paramamosain），紫螯青蟹（S．tranquebarica）和榄绿青蟹（S．olivacea）等4个种。我国以拟穴青蟹占绝对优势，它有个体大、生长快、适应性强、肉味鲜美、营养丰富等特点，已成为具有重要经济价值的海水养殖蟹类之一。     

Morphological deformities in mud crab Scylla serrata juveniles exposed to antibiotics during the larval stage

The effects of antibiotics on the external deformities, growth and survival of mud crab Scylla serrata larvae and juveniles were determined. Zoeae were exposed to oxytetracycline (OTC) (0, 3.0, 6.0, 9.0, 12 mg L^?1) and furazolidone (FZD) (0, 0.5, 1.0, 1.5, 2.0 mg L^?1) in the first and second experiments, respectively, until the late megalopa. The crab instars were grown in nursery tanks for 1 month. Larvae survived until megalopa only at 3.0 and 6.0 mg L^?1 OTC or 0.5 and 1.0 mg L^?1 FZD. These four concentrations were run simultaneously in another experiment. Morphological deformities in zoea 5 were bent dorsal, rostral and furcal spines. There was no significant difference (P > 0.05) on the deformities of zoea 5 in 3.0 and 6.0 mg L^?1 OTC and 0.5 and 1.0 mg L^?1 FZD. Significantly (P < 0.05) higher survival and faster growth were attained in 3.0 mg L^?1 OTC and 0.5 mg L^?1 FZD. Deformities observed in juveniles were fused frontal and lateral spines, asymmetrical and depressed tip of abdominal flap and gap between sternites. High percentage occurrence of deformities was observed in the 6.0 mg L^?1 OTC and 1.0 mg L^?1 FZD in the first and third experiments, respectively. There was no significant difference (P > 0.05) observed in the survival of juveniles in OTC and FZD treatments. However, growth was significantly (P < 0.05) faster in lower concentrations of the two antibiotics. The study shows the effects of OTC and FZD in the morphology of mud crab. Therefore, there is a need to eliminate the use of antibiotics and find alternatives.     

Assessment of dietary lecithin and cholesterol requirements of mud crab, Scylla serrata, megalopa using semi-purified microbound diets

The effects of varying dietary lecithin and cholesterol levels on growth, development and survival of mud crab, Scylla serrata, megalopa were evaluated using six semi‐purified, microbound diets formulated to be iso‐energetic and containing three levels of supplemental lecithin (0, 20 and 40 g kg⁻¹ diet dry weight) and two levels of supplemental cholesterol (0 and 7 g kg⁻¹ diet dry weight). Fifteen megalopa were reared individually in each treatment and the nutritional value of diets was assessed on basis of mean dry weight and mean carapace width of newly settled first crab stage, as well as development time to the first crab stage and overall survival. A significant interaction between supplemental dietary lecithin and supplemental dietary cholesterol was found for final mean dry weight of newly settled crabs, and highest survival (60%) was recorded for megalopa fed diets containing the highest levels of dietary lecithin (39.7–44.1 g kg⁻¹) (diet 5 and 6) regardless of whether diets were supplemented with cholesterol; this rate of survival was identical to that of megalopa fed live Artemia nauplii. The results indicate that supplemental dietary cholesterol may not be essential for mud crab megalopa when fed diets containing sufficient levels of supplemental dietary phospholipids.     

影响锯缘青蟹幼体成活的几个因素

本文对海捕和养殖的锯缘青蟹抱卵及胚胎发育情况进行了观察,并对幼体饵料营养强化、适口性、幼体的培育方式等进行了研究。结果表明：自然诲区捕获的亲蟹其抱卵率和幼体质量要好于人工养殖的亲蟹;胚胎发育的最适盐度为28～33,最适温度为26～30℃;幼体\期和大眼幼体时期摄食的饵料以边缘毛状的活体为佳,便于抱握;涵状幼体、期起应降低苗池中的直射光,投饵多餐少量．增加附着面积．可降低相互残杀率。     

Extension of nursery culture of Scylla serrata (Forsskål) juveniles in net cages and ponds

To address the preference of mud crab farmers for larger size Scylla serrata juveniles (5.0–10 g body weight or BW; 3.0–5.0 cm internal carapace width or ICW), a study was conducted to compare the growth and survival of crab juveniles (2.0–5.0 g BW; 1.0–3.0 cm ICW) produced a month after stocking of megalopae in net cages when reared further in net cages installed in earthen ponds or when stocked directly in earthen ponds. In a 3 × 2 factorial experiment, three stocking densities (1, 3 and 5 ind m⁻²), two types of rearing units (net cages or earthen pond) were used. Megalopae were grown to juvenile stage for 30 days in net cages set inside a 4000 m² brackishwater pond and fed brown mussel (Modiolus metcalfei). Crab juveniles were then transferred to either net cages (mesh size of 1.0 mm) or earthen ponds at three stocking densities. After 1 month, no interaction between stocking density and rearing unit was detected so data were pooled for each stocking density and rearing unit. There were no significant differences in the growth or survival rate of crab juveniles across stocking density treatments. Regardless of stocking density, survival in net cages was higher (77.11±6.62%) than in ponds (40.41±3.59%). Growth, however, was significantly higher for crab juveniles reared in earthen ponds. The range of mean BW of 10.5–16.0 g and an ICW of 3.78–4.33 cm obtained are within the size range preferred by mud crab operators for stocking grow‐out ponds.     

Ontogeny of feeding apparatus and foregut of mud crab Scylla serrata Forsskål larvae

The development of the feeding apparatus of the mud crab Scylla serrata larvae was studied using electron microscopy for mandibles and light microscopy for other paired mouthparts and the foregut. The six paired mouthparts, which consisted of the mandibles, maxillules, maxillae, first maxillipeds, second maxillipeds and third maxillipeds, were dissected from specimens representing each larval stage. The first five paired appendages were already present in newly hatched larvae while third maxillipeds appeared only at the megalopa stage. Mandibles displayed complex incisor and molar processes at early zoeal stages, which became simple in morphology at megalopa. Mandibular palp buds were observed at the zoea 5 stage and these became fully developed as three‐segmented mandibular palps at the megalopa stage. Endopods of other paired mouthparts exhibited increased number of setae and size as the individual metamorphosed from zoeal stages to megalopa and crab instar. The foregut appeared as a continuous cavity at zoea 1 where the cardiopyloric valve was indistinct while the filter gland was clearly identifiable. Zoea 2 and succeeding zoeal stages exhibited a setose foregut; the gastric mill and its lateral and median teeth were prominent at zoea 3 stage. The significance of these morphological changes is discussed in terms of its implication in larval feeding management.     

土霉素在锯缘青蟹体内的药物代谢和消除规律

采用高效液相色谱法检测土霉素,研究土霉素口灌给药途径下在锯缘青蟹体内的药代动力学。锯缘青蟹口灌给药土霉素50 mg/kg后,其血浆、肌肉和肝胰脏中的药峰浓度分别为16.78±1.98 mg/L、9.39±2.12μg/g和32.12±6.12μg/g,达峰时间分别为4 h、8 h和4 h。血浆中土霉素浓度-时间关系曲线符合一级吸收的二室开放动力学模型。土霉素在锯缘青蟹体内分布广泛,其表观分布容积(Vd)为2.129 L/kg;分布半衰期(t1/2α)和消除半衰期(t1/2β)分别为3.200 h和47.856 h,总体清除率(CLs)为0.063 mL/(kg.h)。肌肉和肝胰脏中土霉素浓度与时间关系的药动学参数采用统计矩原理分析,其消除半衰期(t1/2 z)分别为60.145 h和71.009 h,总体清除率(CLz)分别为0.054 g/(kg.h)和0.037 g/(kg.h)。土霉素在精巢和卵巢中达峰时间分别为8 h和12 h,峰浓度分别为9.83μg/g和10.26μg/g。给药后24 d时,血浆、肌肉、肝胰脏、精巢和卵巢中土霉素含量都已低于0.10μg/g。土霉素在锯缘青蟹体内消除比较缓慢。     

Apparent digestibility of selected feed ingredients in diets formulated for the sub-adult mud crab, Scylla paramamosain, in Vietnam

The present study was conducted to explore the potential to incorporate local plant-based feed ingredients into diets formulated for the mud crab species, Scylla paramamosain , commonly exploited for aquaculture in South-east Asia. Four test ingredients (defatted soybean meal, rice bran, cassava meal and corn flour) were incorporated at 30% or 45% inclusion levels in a fishmeal-based reference diet and used in digestibility trials where apparent digestibility coefficients (ADCs) for experimental diets and test ingredients were determined. Generally, high ADC values were obtained using diets containing 30% soybean meal or rice bran. By contrast, the lowest ADC values were obtained for the diet containing 45% cassava meal [70.9% for dry matter (ADMD); 77.1% for crude protein (ACPD) and 80.2% for gross energy (AGED)]. Similar trends were observed when ADC ingredient (I) digestibilities were compared. Specifically, the highest ADCI values were obtained for soybean meal when used at a 30% inclusion level (87.6% ADMDI; 98.4% ACPDI and 95.6% AGEDI) while the lowest ADCI values were obtained using cassava meal at a 45% inclusion level (53.8% ADMDI; 60.2% ACPDI and 67.3% AGEDI). Based on the current findings, we propose that soybean meal and rice bran could be considered for incorporation into formulated diets for S. paramamosain .     

Use of a Durometer to Differentiate Between Soft- and Hard-Shelled Mud Crabs (Scylla serrata)

Commercial mud crab fishers in the Northern Territory, Australia, can harvest both sexes of Scylla serrata year-round. The combination of seasonal moulting and legislation that prohibits the retention of Commercially Unsuitable Crabs (CUCs or “soft crabs”) can result in many individuals being returned to the water at particular times. Concerns regarding consistency in the mandatory manual shell hardness test prompted an evaluation of a durometer as a quantitative assessment tool. Variability in durometer readings from different positions on the exoskeleton made it necessary to impose standardized test sites for each sex: the center of the second abdominal plates for males, and the intersections of perpendicular lines spanning the carapace of females (i.e., a transverse line between the seventh marginal spines and two longitudinal lines originating from each of the third marginal spines). While the vast majority of CUCs returned lower durometer readings than Commercially Acceptable Crabs (CACs), there were some instances where CUCs returned slightly higher readings than CACs (and vice versa). This imperfect matching of hand and durometer test values led to the development of a graphical aid for determining optimal durometer limit values that minimizes the retention and release of CUCs and CACs, respectively.     

Effect of selected feed meals and starches on diet digestibility in the mud crab,Scylla serrata

The present study examined the capacity of the mud crab, Scylla serrata to digest experimental diets that contained different animal and plant‐based feed meals or different levels or types of starch. The apparent dry matter digestibility (ADMD) coefficients for all feed meals tested in the first part of this study, except meat meal, were similar (78–88%). Crude protein digestibility (ACPD) coefficients for all feed meals were relatively high, with values ranging from 86% to 96%. Cotton seed meal, poultry meal, canola meal, fishmeal, soybean meal and lupin meal had similar gross energy digestibility (AGED) values (P>0.05) ranging from 84% to 89%. In the second part of this study, the impact of selected starches on the digestibility of fishmeal‐based formulated diets was assessed. The apparent starch digestibility (ASD) of wheat starch decreased significantly as the inclusion level was increased from 15% to 60%, however, there was no significant effect on ACPD values. At a 30% inclusion level, the ASD of diets containing different starches decreased in the order corn>wheat>potato=rice. Moreover, ACPD values were significantly higher (P<0.05) in the diets containing corn or rice starch than in those containing wheat or potato starches.     

Formalin as an alternative to trifluralin as prophylaxis against fungal infection in mud crab Scylla serrata (Forsskål) larvae

The toxicity of formalin and trifluralin to the larval stages of the mud crab Scylla serrata was compared in a static bioassay. Prophylactic doses of 5, 10, 15, 20 and 25 μg L⁻¹ formalin and 0.05, 0.1, 0.2, 0.4 and 0.8 μg L⁻¹ trifluralin were used. Toxicity was assessed on the basis of survival of larvae after 24, 48, 72 and 96 h exposure to the test chemicals and metamorphosis to the next larval stage. Result shows that larval survival in all stages was significantly reduced at concentrations of 20 and 25 μg L⁻¹ formalin whereas larvae were able to tolerate all trifluralin treatments. However, larvae became more tolerant to high formalin concentrations as the larval stage progressed. Survival was better at 5, 10 and 15 μg L⁻¹ formalin and in all trifluralin treatments than the control in almost all the larval stages. Faster metamorphosis was observed at 5 and 10 μg L⁻¹ formalin and 0.05, 0.1 and 0.2 μg L⁻¹ trifluralin concentrations. Doses of formalin and trifluralin obtained from the toxicity experiments were applied as prophylaxis to newly hatched larvae in white plastic basins. Prophylactic doses of 5 and 10 μg L⁻¹ formalin and 0.05 and 0.1 μg L⁻¹ trifluralin applied every other day were found to be effective in enhancing survival and larval development to megalopa compared with control. However, no megalopae survived to crab instar in all formalin treatments. Although the use of fungicides in rearing systems resulted in higher survival compared with controls, other strategies (i.e. maintenance of good water quality and hygienic practices in the hatchery) should be further investigated as an alternative to the use of chemicals in hatcheries.     

Effects of tank colour on larval survival and development of mud crab Scylla serrata (Forskål)

Hatchery culture of mud crabs has not yet achieved commercial viability despite decades of research efforts. Further research is therefore needed to better understand larval culture requirements of the crab. Based on anecdotal observations, an experiment was carried out to test whether the background colour of the culture vessel affected larval culture success. Newly hatched larvae of Scylla serrata were reared in culture vessels of five colours, i.e., black, dark green, maroon, sky blue and white. Larval survival and development were monitored daily until all of them either moulted to the first crab stage or died. The results showed clear effects of background colour on larval survival. A general tendency of higher larval survival in darker‐coloured backgrounds was evident. In particular, overall zoeal survival for larvae reared in black vessels was significantly higher than those reared in white ones. Background colour also appeared to affect larval development. Larvae reared in darker backgrounds generally had shorter development times and more synchronized moulting. A significant delay in zoeal development was observed in larvae reared in white vessels. Dark backgrounds possibly facilitated more efficient feeding, reduced settlement of larvae at the bottom of the vessels as well as minimized stress. This result appears to be the first to demonstrate that background colour can significantly affect larval survival and development of a crustacean species.     

不同脂肪源饲料对施氏鲟幼鱼生长的影响

在饲料蛋白质水平和能量水平均能满足生长需要的前提下，以不同的油脂配制成6种试验饲料，投喂施氏鲟幼鱼7周，综合分析其生长效率各参数。试验结果表明：混合油组的总体生长效果最好，而葵花籽油和猪油组最差。分析表明：除了ω3多不饱和脂肪酸(PUFA)之外，施氏鲟的必需脂肪酸还同时包括18：2ω6，且施氏鲟对18：3ω3的需求要比18：2ω6大；另外，鱼油氧化后对施氏鲟幼鱼生长也有显著影响。     

锯缘青蟹全人工繁育技术研究

介绍了锯缘青蟹的全人工繁育技术。从亲蟹的强化培育人手提高青蟹抱卵率，2002年青蟹苗种生产中抱卵率为85％．单位水体最高出苗量为7000余只(2茬)／m^3，平均出苗量为1960只／m^3，共育成Ⅱ期幼蟹156．6万只，形成了北方地区锯缘青蟹全人人工工厂化育苗技术的完整体系。