Dietary phospholipids requirement of the early juvenile (C1) swimming crab,Portunus trituberculatus

A 30‐day experiment was conducted to investigate the effect of phospholipids (PLs) on growth performance, survival, fatty acid profile and gene expression of the early juvenile (C1) swimming crab (Portunus trituberculatus). Five semi‐purified diets were formulated with graded PLs levels (0%, 1%, 2%, 4% and 8%). Each diet was fed to four replicates of crabs (30 crabs per replicate, initial weight: 8.4 ± 0.1 mg). In this study, crabs fed diets with 2%, 4% and 8% PLs had significantly higher survival rates than crabs fed with 0% and 1% PLs. Crabs fed diets with 0% and 1% PLs had significant lower weight gain (WG) than crabs fed with 8% PLs. But crabs fed with diets containing 2%, 4% and 8% PLs showed no significant difference in WG. Besides, the content of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in whole body was higher than that in diets, but the monounsaturated fatty acids (MUFA) content showed an opposite trend. The EPA and DHA content in whole body increased with the increase in dietary PLs. Moreover, the moulting number increased with the increasing dietary PLs levels, the lowest moulting number was observed in crabs fed diets without supplementation of PLs. The ecdysteroid receptor (EcR) was found to be significantly up‐regulated by PLs levels. This study demonstrated that 2% PLs could meet the requirement of early juvenile (C1) swimming crab based on the growth and survival. And 4%–8% PLs supplemented in diets could reduce the leaching of feed.     

Effects of dietary phospholipid and cholesterol levels on growth and fatty acid composition of juvenile swimming crab,Portunus trituberculatus

The objective of this study was to evaluate the effects of different phospholipids (PL) and cholesterol (CH) levels on the growth, moulting and fatty acid composition of juvenile swimming crab, Portunus trituberculatus. Six diets were designed to contain three PL levels (0, 10 and 20 g/kg) and two CH levels (2 and 8 g/kg). Juvenile swimming crabs (3.48 ± 0.02 g/crab) were reared for 8 weeks. The weight gain (WG) was significantly (p < .05) increased by supplementation of 8 g CH/kg of diet. However, no significant interaction between dietary PL and CH levels was found on the growth performance (p > .05). The moulting frequency (MF), protein efficiency ratio (PER) and feed efficiency ratio (FCR) were not significantly (p > .05) affected by the dietary treatments. The serum total cholesterol (TCH) significantly (p > .05) increased with increasing dietary PL level. The C20:4n‐6 and C20:5n‐3 content of the whole body of crabs increased with the addition of PL to the diet containing 2 g/kg CH of diet. An interaction was observed between PL and CH on certain saturated and unsaturated fatty acid concentrations of body.     

Changes to the histological gill structure and haemolymph composition of early blue swimmer crab Portunus pelagicus juveniles during elevated ammonia‐N exposure and the post‐exposure recovery

It is yet unclear whether sub‐lethal ammonia‐N levels cause irreparable damage to aquatic crustaceans, or if recovery is possible, the potential factors involved. The aim was to investigate the effect of 0.706 and 2.798 mmol L^?1 ammonia‐N exposure on the haemolymph osmolality, Na⁺, K⁺, Ca²⁺, pH, ammonia‐N, total haemocyte counts (THC) and gill histopathology of Portunus pelagicus juveniles at 0, 3, 6, 12, 24 and 48 h respectively. Following 48 h, crabs were transferred to pristine seawater allowing a recovery period up to 96 h and similarly measured. In addition moribund crabs, induced from lethal ammonia‐N levels of 7.036 and 10.518 mmol L^?1, were measured for haemolymph osmolality/ions and pH levels. The results demonstrate that despite severe gill damage within 6‐ and 1 h of 0.706 and 2.798 mmol L^?1 ammonia‐N exposure, respectively, no significant change (P>0.05) in the haemolymph osmolality, Na⁺, K⁺, Ca²⁺ or pH levels occurred or by ammonia‐N‐induced morbidity. Although the gills can completely recover within 24 and 48 h post exposure to 0.706 and 2.798 mmol L^?1 ammonia‐N, respectively, likely facilitated by significant haemocyte increases (P<0.05) within the haemolymph and gill lamellae, dependent factors were the previous ammonia‐N concentration and recovery duration while individual variability was also noticed.     

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%).     

Effect of dietary lipid levels on growth performance,fatty acid profile and enzymatic activity of juvenile swimming crab,Portunus trituberculatus

The main objective of this investigation was to study the lipid requirements of the early juvenile (C1) swimming crab (Portunus trituberculatus) based on growth performance, survival, moulting and fatty acid profile. Four test diets were formulated with graded lipid levels (3.63%, 6.70%, 10.72% and 13.91%). Each diet was fed to 4 replicates of crabs (30 crabs per replicate initial weight (8.4 ± 0.1 mg). In this study, crabs fed diets with 13.91% lipid had significantly (p < 0.05) higher survival than crabs fed with 3.63% lipid, but no significant (p > 0.05) improvement of survival was observed when dietary lipid increase from 6.70% to 13.91%. Crabs fed diets with 10.72% and 13.91% lipid had significant higher weight gain(WG) than crabs fed with 3.63% lipid. While crabs fed with diets containing 6.70%, 10.72% and 13.91% lipid showed no significant (p > 0.05) difference in weight gain(WG). Moreover, the lowest moulting number was observed in crabs fed diets with 3.63% lipid, but there was no significant difference (p > 0.05) among other groups. The content of LC‐PUFA and DHA in the crabs fed diets with 3.63% lipid was significant (p < 0.05) lower compared to other groups. But there was no significant (p > 0.05) difference in EPA and ARA content among all groups. The activity of lipase increased as dietary lipid level increased (from 6.70% to 10.72%). However, beyond 10.72%, a significant (p < 0.05) decreased in lipase activity was observed. The regression analysis of weight gain data indicated that crab fed diet containing 10.47% lipid level is considered as optimum lipid level for its maximum growth and moulting process.     

Effects of phospholipid addition to diets with different inclusion levels of fish oil on growth and fatty acid body composition of juvenile swimming crab Portunus trituberculatus

Six experimental diets were designed with two phospholipid (PL; 0% and 1.5%) and three fish oil levels (0%, 1% and 3%) to evaluate the effects of dietary fish oil and PL levels on growth, survival and fatty acid composition of juvenile swimming crab, Portunus trituberculatus. Diets were iso‐energetic and iso‐nitrogenous and each diet was fed to triplicate groups (initially weight, 24.88 ± 0.04 g per crab) for 59 days. Weight gain (WG) and specific growth rate (SGR) increased with dietary PL addition to 0% fish oil‐supplemented diets (P < 0.05). On the other hand, WG and SGR decreased with dietary PL addition to 3% fish oil diets (P < 0.05). Crabs fed PL supplemented diets had higher haemolymph low‐density lipoprotein cholesterol concentrations and muscle crude lipid levels (P < 0.05) than crabs fed a none PL supplemented diet. The percentage of highly unsaturated fatty acids (HUFA; % total FA) in both polar and neutral lipids fractions of muscle tissue only increased in case of PL addition to 0% and 1% fish oil‐supplemented diets (P < 0.05). HUFA levels in the neutral lipids fraction of the hepatopancreas increased by dietary PL addition at each dietary fish oil level (P < 0.05). In this study, both dietary fish oil and PL addition contributed to a high n‐3/n‐6 ratio in muscle and hepatopancreas of P. trituberculatus. In conclusion, PL addition is only meaningful with fish oil‐deficient diets, in which case it enhanced lipid transport and HUFA absorption efficiency, hence improving the nutritional value of the diet.     

Effects of dietary DHA/EPA ratios on growth performance,survival and fatty acid composition of juvenile swimming crab (Portunus trituberculatus)

This study was conducted to investigate the effects of dietary docosahexaenoic to eicosapentaenoic acid (DHA/EPA) ratios on growth performance, survival and fatty acid composition of juvenile swimming crab (Portunus trituberculatus). Four isonitrogenous and isoenergetic experimental diets were formulated to contain different DHA/EPA ratios (0.70, 0.84, 1.06 and 1.25). There were three replicates (15 crabs per replicate) for each diet treatment. The crabs were fed (about 6–8% body weight) twice daily for 8 weeks. A good growth performance and feed utilization were observed in swimming crabs fed the diets with DHA/EPA ratios of 0.70 and 0.84. Crabs fed diet with 0.70 DHA/EPA ratio showed a significantly higher weight gain (WG) compared with the crabs fed the diet with DHA/EPA ratio of 1.25 (P < 0.05). The result of this study showed that the survival increased with decreasing the ratio of DHA/EPA. The DHA/EPA ratios in polar lipid from tissues were influenced by the dietary DHA/EPA ratios. Results of this study indicated that the growth performance and survival of juvenile swimming crab are correlated to the DHA/EPA ratio in the diets, and the best growth performance and survival were achieved with the ratios of 0.70–0.84.     

Use of escape vents to improve size and species selectivity of collapsible pot for blue swimming crab <Emphasis Type="Italic">Portunus pelagicus</Emphasis> in Thailand

Laboratory experiments were firstly conducted to determine the possibility of modification of the collapsible pot for blue swimming crab Portunus pelagicus by designing appropriate escape vents in terms of shape, position, and size to improve size selectivity by reducing the catch of immature crabs. In laboratory observations, nearly square-shaped vents of 35 mm height and 45 mm length, located at the lower part of the side panel, showed the best performance to allow the escape of immature-size crabs by side-crawling escape behavior through the vents, with L _50% selection carapace length (CL) of 39.1, 44.4, and 48.7 mm CL for 40, 45, and 50 mm vent length, respectively. Comparative fishing trials between conventional and vented pots with vents of 35 × 45 mm size were conducted to examine the crab size and species selectivity. Use of the vented pots was found to reduce the number of immature crabs in the catch from 70.5% to 11.0% on average, while not affecting the catch efficiency of mature-size crabs. The vented pots also had a selective effect for reducing the catch of bycatch species in comparative fishing trials.     

Effects of different dietary vitamin E and astaxanthin levels on growth,fatty acid composition and coloration of the juvenile swimming crab,Portunus trituberculatus

Six isonitrogenous and isolipidic diets were formulated containing two astaxanthin (AX; 0 and 90 mg/kg) and three vitamin E (VE) levels (0, 25 and 50 mg/kg). There were three replicates (18 crabs per replicate) for each treatment. Juvenile swimming crab (initial weight 31.65 ± 0.06 g/crab) were fed different diets for 8 weeks. After the feeding trial, growth performance was not significantly affected by the different treatments. Crabs fed with AX‐supplemented diets showed more redness. Whole body 22:6n‐3, 22:4n‐6 and 20:5n‐3 levels increased with the dietary addition of AX (P = .009, P = .002 and p = .042, respectively). The malondialdehyde (MDA) concentrations of fresh/frozen hepatopancreas and frozen muscle were significantly decreased by the dietary AX supplementation (p < .001, p = .010 and p = 0.011, respectively). These findings provide evidence that dietary AX has an ability to improve the redness of the shell and reduce the MDA concentrations of tissues. Furthermore, there is no strong interactive relationship between dietary VE and AX on the coloration and fatty acid concentrations for the swimming crab.     

Ovarian development and induced oviposition of the overwintering swimming crab Portunus trituberculatus (Brachyura: Portunidae) reared in the laboratory

ABSTRACT:   Females of the swimming crab Portunus trituberculatus were cultured under natural temperature and high temperature (21°C) conditions to examine ovarian development and oviposition from autumn (October) to the beginning of breeding season in the following spring (May). Ovaries developed because of vitellogenesis of oocytes from October to December and their developmental state did not change during the overwintering period from December to March. In spring, from late March to mid May, some ovaries reached prematuration and maturation stages and ovipositions began from mid–late April under natural temperature conditions. Females reared in tanks of high temperature regimes oviposited earlier than those reared in tanks of natural temperatures. The number of days to oviposition tended to decrease with advancement of the beginning times (early February to late April) of culture of females under high temperature regimes; induced ovipositions were achieved more easily with increasing photoperiod from c . 12 h in early February to c . 14 h in late April. Therefore, it is inferred that the temperature and photoperiod are important environmental factors controlling ovarian development and oviposition.     

iTRAQ‐based identification of differentially expressed proteins related to growth in the swimming crab,Portunus trituberculatus

The swimming crab, Portunus trituberculatus, is an important farmed species in China, and it has been studied extensively, which requires more information on its genetic background. To date, information on the proteomics of P. trituberculatus is scarce. In this study, we used isobaric tags for relative and absolute quantitation (iTRAQ)‐based proteomics to investigate growth regulation in P. trituberculatus. Total proteins were isolated from five tissues (eyestalk, gill, heart, hepatopancreas and muscle). Equal quantities of protein from each tissue were pooled at the proteome level using the iTRAQ method. A total of 961 proteins were identified using liquid chromatography coupled with tandem mass spectrometry and de novo sequencing data. Using a 1.2‐fold change in expression as a physiologically significant benchmark, 30 differentially expressed proteins (DEPs) were found to undergo differential expression related to the growth of P. trituberculatus, and most of the growth‐related proteins, including those involved in metabolism, immune responses, DNA duplication and protein synthesis, were upregulated, indicating that conservation of energy is an important strategy to cope with growth. There was a high consistency between the expression levels determined using iTRAQ and mRNA, highlighting the high reproducibility of our proteomic approach and its great value in elucidating the molecular mechanisms of P. trituberculatus.     

三疣梭子蟹幼蟹配合饲料中蛋白质、脂肪、粗纤维适宜含量研究

以初始平均体重为7.1 g的三疣梭子蟹(Portunus trituberculatus)幼蟹为试验对象,以粗蛋白、粗脂肪和粗纤维为试验因素,各设3个水平,采用正交试验设计L9(34),配制成9种饲料,在室内水族箱进行为期4周的饲养试验,以探讨三疣梭子蟹幼蟹配合饲料中蛋白质、脂肪和粗纤维的适宜含量.结果表明,三疣梭子蟹幼蟹配合饲料中蛋白质、脂肪和粗纤维的适宜含量分别为40%、6%～8%和≤5%时,三疣梭子蟹幼蟹增重率和蜕壳率高,饲料系数低和蛋白质效率较高.     

Long‐term low‐salinity stress affects growth,survival and osmotic related gamma‐aminobutyric acid regulation in the swimming crab Portunus trituberculatus

A 30‐day experiment was conducted to investigate the effects of long‐term low‐salinity stress on the growth performance and osmotic related chlorine ion channel ligand regulation: gamma‐aminobutyric acid (GABA) content and GABA_A receptor‐associated protein (GABARAP) expression in Portunus trituberculatus. The salinity levels of both the control group and the experimental group were 30 and 12 psu respectively. After rearing for 30 days, the specific growth rate and survival rate were compared between the two groups, and salinity 6 psu was used to test the salinity tolerance. The results were as follows: (a) Both the specific growth rate and survival rate were significant lower in the experimental group (p < 0.05) after 30 days; (b) After challenge with salinity 6 psu for 72 hr, the crabs of experimental group had a 100% survival rate, whereas the crabs of the control group were all dead within 48 hr; (c) The content of GABA and the gene expression level of GABARAP in experimental group were significant different from control group (p < 0.05) after challenge via salinity 6 psu. In the control group, the GABA content increased rapidly from 9.96 ± 2.09 to 42.00 ± 5.94 µg/g; however, in the experimental group, it only increased to 27.82 ± 2.55 µg/g; the gene expression of GABARAP in the experimental group increased to the maximum at 24 hr, then decreased and stabilized at 48 hr, suggesting that GABA and GABARAP were trigged during the early stage of low‐salinity stress resistance.     

Isolation,characterization and virulence of Mesanophrys sp. (Ciliophora: Orchitophryidae) in farmed swimming crab (Portunus trituberculatus) in eastern China

A disease outbreak occurred in swimming crab (Portunus trituberculatus) farmed in eastern China, with a mortality rate of more than 80%. To further investigate the characteristics and pathogenesis, we reported isolation, characterization and virulence of the causative agent of this disease from 10 sick crabs. Histopathological observation found that multiple tissues, especially haemolymph, contained lots of ciliates. The ciliate was isolated and cultured in vitro, and molecular and morphological studies were done. The results showed that SSU rDNA and LSU rDNA sequences of the ciliate were similar to Mesanophrys ciliates (>96.81%), while ITS1-5.8s-ITS2 sequence was similar to Mesanophrys pugettensis (95.37%) and identical to Orchitophrya stellarum (100%). Furthermore, the results of the morphological study confirmed that the ciliate was similar to Mesanophrys ciliates and O. stellarum cultured in supportive media, but different from O. stellarum cultured in living sperm cells of starfish (Leptasterias spp.). Also, the growth of the ciliate did not interfere with light, which was different from O. stellarum. Accordingly, the ciliate was classified as genus Mesanophrys and temporarily named as Mesanophrys sp. In addition, experimental infection confirmed that Mesanophrys sp. was the pathogen that infected farmed crabs. In summary, Mesanophrys sp. was first isolated and characterized in P. trituberculatus.     

Effect of salinity on the rearing performance of juvenile golden pompano Trachinotus ovatus (Linnaeus 1758)

Effect of rearing salinity on the performance of juvenile golden pompano Trachinotus ovatus (Linnaeus 1758) was studied under a laboratory condition. Fish growth, survival, RNA/DNA ratio, pepsin activity, α‐amylase activity and FCR were used as evaluation criteria. The growth and RNA/DNA ratio were significantly affected by the rearing salinity. High growth rate and RNA/DNA ratio were observed when fish were reared at the salinity of 34‰. The pepsin activity of fish was not significantly affected by the rearing salinity. However, the α‐amylase activity of fish was significantly affected by the rearing salinity. The α‐amylase activity of fish reared at the salinity of 10‰ was significantly lower than fish cultured at the salinity of 34‰. Rearing salinity can significantly affect the FCR of juvenile golden pompano. The FCR of fish cultured at the salinity of 10‰ was 5‐times higher than the FCR of fish reared at 34‰. Results from the present study indicate that juvenile golden pompano can be reared above 26‰ without affecting fish performance, and the salinity of 10‰ may be too low to rear juvenile golden pompano as fish growth, RNA/DNA ratio and α‐amylase activity were reduced.     

饲料中蛋白水平对三疣梭子蟹雌体生长、卵巢发育和生化组成的影响

本研究配制蛋白水平分别为32.16％、36.13％、39.59％和41.24％的等脂等能配合饲料(分别记为饲料1～4),对初始体重为(10.98±0.28)g的三疣梭子蟹(Portunus trituberculatus)雌体进行池塘养殖120 d,以探究饲料蛋白水平对三疣梭子蟹雌体生长、卵巢发育和生化组成的影响.结果...     

三疣梭子蟹细胞Cdk7基因克隆及其在卵巢发育中的表达

本实验应用RACE技术克隆获得了三疣梭子蟹(Portunus trituberculatus)周期蛋白依赖性激酶7(Cdk7)基因c DNA序列全长。基因5′和3′非编码区域(UTR)以及开放阅读框的长度分别是23 bp、178 bp和1056 bp,预测编码一个含有351个氨基酸,分子量为39.91 k D的蛋白质,包含一个丝氨酸/苏氨酸催化保守结构域和T-loop结构。同源性分析表明,三疣梭子蟹CDK7氨基酸序列与其他物种Cdk7相似度较高,说明Cdk7基因具有很好的保守性。实时荧光定量PCR结果显示,三疣梭子蟹Cdk7基因在卵巢组织中的表达量显著高于其他组织(P0.05)。Cdk7基因在卵巢发育不同时期的表达量存在差异,其在I期和Ⅱ期的表达量显著高于其他时期(P0.05)。去除眼柄后该基因在卵巢组织中的表达呈现先升高后降低的趋势,并于第4天达到最大值。本研究的结果表明,Cdk7基因参与了三疣梭子蟹卵巢发育调控,为深入开展三疣梭子蟹和其他甲壳动物性腺发育调控机理研究提供了重要信息。     

First results of embryonic development,spawning and larval rearing of the Mediterranean spider crab Maja squinado (Herbst ) under laboratory conditions,a candidate species for a restocking program

The Mediterranean spider crab, Maja squinado, is depleted due to overfishing. The crab has virtually disappeared from areas where it was abundant, such as the Balearic Islands and the Catalan coast. Maja squinado, is economically and ecologically very valuable, and it is essential to obtain information on its biology and rearing conditions to attempt to repopulate the damaged stocks of the species in the Mediterranean basin. Herein, we describe the first successful rearing of M. squinado under laboratory conditions. Our results show that M. squinado is an excellent candidate for restocking using cultured juveniles. Two consecutive broods with a 1–4 day interbrood period were observed in the laboratory in wild‐caught females, the maximum observed duration of embryonic development of the egg mass being 32 days at 18.4 ± 0.9°C, and went through four different stages. The complete larval and first juvenile development was studied in laboratory cultures fed enriched Artemia metanauplius. At 19.6 ± 0.6°C, development from hatching to first crab moult took 17 days, and it comprised two zoeae stages and one megalopa stage. The survival rate at the different stages was monitored, and 7.13 ± 2.3% was achieved at the first crab instar.     

Intensive rearing of juvenile crayfish (Pacifastacus leniusculus,Astacidae) during the first 6 months: effects of size grading

Recent advances in crayfish feeding have enabled the development of size grading studies from the start of first‐feeding. A 180‐day experiment aimed at intensive rearing of Pacifastacus leniusculus was carried out under controlled conditions, evaluating the effects of size grading at two different periods from the onset of exogenous feeding. Stage 2 juveniles were stocked in fibreglass tanks at a density of 100 m², and fed a dry diet for salmonids combined with restricted amounts of Artemia cysts. Five groups were tested: no grading, grading at 60 days (large and small size) and grading at 100 days (large and small size). After 6 months, no significant differences were found in the survival among groups (mean: 73.06%). The highest final growth (pooled results from upper and lower classes: 17.39 mm carapace length, 1.43 g weight) was achieved by the crayfish sorted at 60 days, showing significant differences from the ungraded group. Smaller crayfish graded at 60 days grew significantly faster than smaller crayfish graded at 100 days. The food conversion ratio was lower in the graded groups (mean: 2.64), showing significant differences from the ungraded group (3.23). This study shows that size grading allows a better performance and an improved feeding efficiency.