Reproductive performance and offspring quality of Chinese mitten crab Eriocheir sinensis (H. Milne-Edwards) females fed an optimized formulated diet and the razor clam Sinonovacula constricta

After feeding female Eriocheir sinensis on an optimized formulated diet or fresh razor clam Sinonovacula constricta for 7 months, their reproductive performance and offspring quality were compared. To evaluate diet nutrient contents, the proximate, fatty acid and amino acid compositions of the formulated diet and the razor clam were analysed. The nutritional value of the diets was determined by assessing survival, gonadosomatic index (GSI) and hepatosomatic index (HSI) of female crabs from both diet treatments, together with the percentage of females that spawned, total egg production per female and fecundity (number of eggs g^?1 female wet weight). Furthermore, the quality of eggs and newly hatched larvae from the two dietary treatments were determined using the following parameters: egg diameter, wet weight and dry weight, hatchability, proximate and fatty acid profile of eggs, larval carapace length, resistant to starvation and osmotic shock, larval survival and development to the zoea II stage. Higher protein, phospholipids (PL) and amino acids (AA) contents were found in the razor clam while the formulated diet contains higher levels of ash, total lipid (TL) and 18:1n‐9, 18:2n‐6 and 22:6n‐3 fatty acids. Although female crabs fed the two different diets showed similar reproductive performances, newly hatched zoea I larvae produced by the crabs fed the formulated diet had significantly longer mean carapace length and shorter development time to the zoea II stage under identical culture condition (P<0.05). Moreover, dietary fatty acid appeared to have more significant effects on the fatty acid composition of the hepatopancreas than it did on mature ovaries or eggs. This suggests that the fatty acid profile of mature ovaries is indicative of the specific fatty acid required for ovarian development in E. sinensis. In conclusion, our results show that the optimized formulated diet developed in this laboratory can totally replace the razor clam, a broodstock food widely used in E. sinensis hatcheries in China. This encouraging result should facilitate more reliable hatchery production of this important aquaculture species.     

Effects of dietary phospholipids and highly unsaturated fatty acids on the precocity,survival, growth and hepatic lipid composition of juvenile Chinese mitten crab,Eriocheir sinensis (H. Milne‐Edwards)

Precocious puberty is one of the major constraints to the further development of Chinese mitten crab (Eriocheir sinensis) farming industry. Although dietary phospholipids (PL) and highly unsaturated fatty acids (HUFA) supplementation have been shown to enhance the growth of larval E. sinensis in other studies, it is still unknown whether this also leads to a higher precocity rate for juvenile E. sinensis. This study was conducted to investigate the effects of dietary PL and HUFA on precocity, survival, growth and hepatic lipid composition of juvenile E. sinensis. Two diets were formulated with PL [3.95% dry weight (DW)] and HUFA (0.98% DW) supplementation (diet A) and without PL and HUFA supplementation (diet B) and fed to juvenile E. sinensis. Although dietary PL and HUFA levels did not significantly affect the survival and growth performance of juvenile E. sinensis, compared with crabs fed diet A, a higher precocity rate was found among juvenile E. sinensis fed diet B (P=0.051). A higher total lipid content, but significantly lower levels of HUFA and PL (P<0.05) were found in the hepatopancreas of crabs fed diet B than in those fed diet A. Meanwhile, the precocious females had significantly lower hepatosomatic index, arachidonic acid (20:4n‐6), eicosapentaenoic acid (20:5n‐3) and docosahexaenoic acid (22:6n‐3) contents in their hepatopancreas when compared with that of the normal immature juveniles (P<0.05). The results suggest that the occurrence of precocious puberty among farmed juvenile E. sinensis could be reduced by the inclusion of appropriate level of dietary PL and HUFA.     

pH胁迫对中华绒螯蟹5种非特异性免疫指标的影响

测定了不同pH(4.5,6.0,7.5,9.0,10.5)胁迫1、4、7、10d后中华绒螯蟹(Eriocheir sinensis)血细胞呼吸爆发(RP)、血细胞破碎物上清液(HLS)酚氧化酶(PO)、血清溶菌酶(LSZ)、酸性磷酸酶(ACP)及碱性磷酸酶(ALP)活性的变化.结果表明:在pH胁迫后第1天,pH4.5组...     

Effect of dietary soybean lecithin on reproductive performance of Chinese mitten crab Eriocheir sinensis (H. Milne-Edwards) Broodstock

The effect of increasing levels of dietary phospholipids (PL) on the ovarian development and reproductive performance of Chinese mitten crab Eriocheir sinensis were investigated using four semipurified formulated diets supplemented with 0%, 1.2%, 2.4%, and 3.6% PL. Four groups of 40 females, with an average individual body weight of 95–120 g, were fed the experimental diets for a period of 7 months. Male crabs were introduced into the female rearing system in March, and mating, spawning, and egg hatching occurred in the following month. After 10 weeks of feeding, females fed the diet with 2.4% PL had a significantly higher gonadosomatic index (GSI) than females fed the diet with 0% PL, whereas females fed the diet with 1.2% PL had a significantly higher hepatic moisture content and lower hepatic lipid content than the other groups (P < 0.05). After mating, the spawning rate was higher among females fed diets with 3.6% and 2.4% PL (95% and 92%, respectively) compared with females fed diets with 0% and 1.2% PL (both 81%). The results showed that egg production (total number of eggs/female) and fecundity increased with increasing dietary PL level, with females fed diets with 0% and 1.2% PL supplementation having significantly lower values than females fed the diet with 3.6% PL (P < 0.05). In conclusion, our results suggest that dietary PL supplementation has a positive effect on ovarian development and reproductive performance of E. sinensis broodstock. Further study should aim to investigate the optimal PL level in the broodstock diet of E. sinensis in respect to offspring quality.     

Effects of dietary phospholipid and highly unsaturated fatty acid on the gonadal development, tissue proximate composition, lipid class and fatty acid composition of precocious Chinese mitten crab, Eriocheir sinensis

Precocious puberty is a major constraint to the economical rearing Eriocheir sinensis farming. Although dietary phospholipid (PL) and highly unsaturated fatty acids (HUFA) reportedly enhanced ovarian development in normal adult E. sinensis , it had opposite effects of reducing precocity in juveniles . This study investigated the effects of dietary PL and HUFA on survival, gonadal development and biochemical composition of precocious E. sinensis . Two diets with PL and HUFA supplementation (diet A) and deficient (diet B) were formulated and fed to precocious E. sinensis . Although no significant differences were found on survival and gonadosomatic index of crabs fed both diets, crabs fed diet B had significantly higher hepatosomatic index and hepatopancrean lipid content than crabs fed diet A. The percentages, 14:0, 18:1n-9, 20:5n-3 and 22:6n-3, in hepatopancreas, muscle and gonads were positively correlated to the diets. Interestingly, the crabs fed diet B accumulated significantly higher 18:2n-6 in gonads and muscle than those fed diet A. Furthermore, substantially higher 20:4n-6 was detected in the gonad and muscle than in the diets. These results suggested that dietary PL and HUFA had limited effects on male gonad biochemical composition. Meanwhile, the precocious E. sinensis may be capable of negating the negative effects of HUFA deficiency by enhanced tissue 18:2n-6 accumulation and converting 18:2n-6 to 20:4n-6.     

超低温冷冻对中华绒螯蟹胚胎线粒体DNA的影响

为了解超低温冷冻对中华绒螯蟹(Eriocheir sinensis)胚胎线粒体DNA影响,选取细胞分裂期胚胎为研究对象,研究了4种玻璃化液处理和超低温冷冻对胚胎线粒体DNA的影响。结果表明:经过玻璃化液处理和冷冻后,胚胎线粒体上的Cytb基因能被稳定扩增,PCR电泳产物大小约1 600 bp,产物经回收后进行双向测序、比对和校正,获得10条Cytb基因全序列,长度为1 135 bp,其中有5个核苷酸变异位点,1 135 bp的Cytb基因序列一共编码378个氨基酸残基,从ATG第一位起始密码子开始到1 140位终止。经过玻璃化液处理和冷冻后,胚胎线粒体上的COI基因也能被稳定扩增,PCR电泳产物大小约1 600 bp,产物经回收后进行双向测序、比对和校正,获得10条COI基因全序列,长度为1 534 bp,其中有6个核苷酸变异位点,1 534 bp的COI基因序列一共编码511个氨基酸残基,从ATG第一位密码子开始到1 533位终止。经过玻璃化液处理和冷冻后,中华绒螯蟹胚胎线粒体上Cytb和COI基因共有11个位点发生了变异,但不同的玻璃化液处理和冷冻对碱基变异没有影响,发生的碱基变异中存在4种类型,即G→A、A→G、C→T、T→C,变异间只有转换发生,没有颠换发生,这些变异均未引起相应氨基酸残基的变异。     

Ingestion of Artemia nauplii by Chinese mitten crab Eriocheir sinensis zoea larvae

A series of ingestion trials were conducted to determine the ingestion rate of Artemia nauplii by Eriocheir sinensis zoea larvae with increasing densities of Artemia and with or without rotifers as a co-feed. At each zoeal stage, 10 groups of 10 larvae were reared individually in glass beakers and fed with increasing densities of newly hatched Artemia nauplii (0.5, 2.5, 5, 10 and 20 individual (ind.) mL⁻¹) with or without rotifers (15–25 ind. mL⁻¹) as a co-feed. The average number of ingested Artemia was measured over 24 h. In addition, the average larval development rate (Larval Stage Index, LSI) over a longer period (time needed for the best treatment to reach 100% moult or metamorphosis to the next larval stage) was compared. The results showed that Artemia ingestion rate of E. sinensis larvae increased with increasing prey densities and larval development, and had a significantly negative correlation with rotifer consumption for all zoeal stages. Rotifers as an alternative prey significantly affected the intake of Artemia at early larval stages (Z1 and Z2) and promoted LSI at a lower Artemia density. Further experiments are needed to clarify the effect of prey density on survival and larval development when larvae are reared communally.     

磷脂和HUFA对中华绒鳌蟹幼蟹存活、生长、蜕壳及生化组成的影响

实验用中华绒鳌蟹(Eriocheir sinensis)幼蟹初始体质量为1.8 g左右.饲料A添加6%鱼肝油和4%大豆卵鳞脂(饲料含4.53%磷脂和1.08%HUFA),饲料B添加10%的猪油(饲料含0.39%磷脂和0.18%HUFA);实验蟹单个体饲养(可防止蜕壳期间蟹相互残杀而获得饲料外的营养源),雌蟹(F)、雄蟹(Lul)各60个,实验周期120 d.结果表明:两组饲料条件下,幼蟹的存活率、增重率、特定生长率、平均蜕壳次数、蜕壳间隔时间及肝胰腺指数均无显著差异(P>0.05).但无论是雌蟹还是雄蟹,A组幼蟹肝胰腺和肌肉的总脂含量显著低于B组(P<0.05),并且A组蟹肝胰腺和肌肉中磷脂、多不饱和脂肪酸和高度不饱和脂肪酸的含量显著高于B组(P<0.05).这表明,在中华绒鳌蟹幼蟹饲料中添加4.53%磷脂和1.08%HUFA对其存活、生长、蜕壳无显著影响,但可使其肝胰腺和肌肉中的总脂含量降低,而磷脂、多不饱和脂肪酸和高度不饱和脂肪酸的含量升高.本研究结果旨在为中华绒鳌蟹幼蟹的营养学研究提供理论依据,同时也为生产上幼蟹饲料的开发提供参考.     

Ovarian re‐maturation following the first spawning in the Chinese mitten crab,Eriocheir sinensis (H. Milne‐Edwards)

The present study investigated the external and histological changes in the ovary and measured the gonadosomatic index (GSI) and hepatosomatic index (HSI) during the re‐maturation of the Chinese mitten crab Eriocheir sinensis. The results show that the ovarian re‐maturation cycle of this crab species can be divided into four stages, i.e. Stage I: the ovary had an X shape and appeared milky white or with a mottled buff colour, and the dominant gametocyte at the stage were oogonia (OG, 28.3%) and previtellogenic oocytes (PR, 31.6%); Stage II: the ovary appeared light yellow or tan in colour, dominated by endogenous vitellogeic oocytes (EN, 69.3%). The yolk globules of unreleased mature oocytes (MO) from the previous spawn were absorbed, leaving many vacuoles in the retrogressing MO; Stage III: the ovary appeared crimson red or deep purple, and although the main type of gametocyte was exogenous vitellogenic oocytes (EX), near‐mature oocytes (NO) could also be found in the late phase of the ovarian development stage; Stage IV: the ovary was ripe and filled with MO that contained large yolk globules. During the period of the second ovarian maturation cycle, the GSI increased significantly from the time just after the first spawning (P<0.01). However, although HSI appeared to decrease within days after spawning (DAS), no significant correlation was found between GSI and HSI or between HSI and DAS (P>0.05). Furthermore, the final GSI and the volume of MO of E. sinensis during the second maturation cycle were drastically lower than those of the first maturation cycle.     

Purification and characterization of a new reovirus from the Chinese mitten crab, Eriocheir sinensis

A new reovirus was recently isolated from a freshwater crab, the Chinese mitten crab, Eriocheir sinensis, in China. The complete viral particles are 55 nm in diameter, icosahedral, non-enveloped and have a mean buoyant density of 1.39 g cm(-3) in CsCl gradient. The viral genome is composed of 12 pieces of dsRNA with an electrophoretic pattern of 3/4/2/3. This virus infects connective tissue of the gills, gut and hepatopancreas. Partial cDNA cloning and sequence analysis showed that the RNA-dependent RNA polymerase is located in the first RNA segment. From its biochemical, ultrastructural and physicochemical properties, this virus is quite different from the genus Aquareovirus (Reoviridae). It may represent a new genus of Reoviridae, different from the other crab reoviruses, P and W(2).     

Kinetic analysis of oxytetracycline residues in Chinese mitten crab,Eriocheir sinensis,muscle following intramuscular administration

Crab culture is a very important economic industry in China. An epidemic of tremor disease of Chinese mitten crabs, Eriocheir sinensis, has become a serious problem in recent years. A spiroplasm has been proved to be the causative agent of this disease. Oxytetracycline (OTC) is used widely in aquaculture and was confirmed to be very effective against this pathogen. In this study, the distribution and depletion patterns of OTC in crab muscle were evaluated following single intramuscular doses of 2, 8 and 40 mg kg^?1 body weight. OTC was detected with a validated HPLC method. Concentration–time profiles were well described by a three‐compartment model with first‐order absorption after a single dose of 8 and 40 mg kg^?1. For comparison, a non‐compartment model was employed. A withdrawal time of 48.29 and 55.92 days was suggested prior to consumption after receiving 8 and 40 mg kg^?1. A recommended therapeutic dose of OTC in theory was calculated to be 36.37 mg kg^?1. OTC was distributed well throughout the body. The elimination of OTC in muscle was slower compared with fish and other crustaceans. A dose of 40 mg kg^?1 is suggested for practical use.     

中华绒螯蟹线粒体DNA l2S rRNA基因片段的序列研究

首次进行了中华绒螯蟹（ Ｅｒｉｏｃｈｅｉｒ ｓｉｎｅｎｓｉｓ）线粒体 ＤＮＡ １２ＳｒＲＮＡ的序列分析。参考果蝇、蚤状溞序列对中华绒螯蟹线粒体ＤＮＡ １２ＳｒＲＮＡ基因片段做了引物设计、ＰＣＲ扩增及序列测定,结果得到 ４５７ ｂｐ的碱基序列,其Ａ、Ｔ、Ｇ、Ｃ含量分别为 １５９ｂｐ（３４．７９％）、１７７ｂｐ（３８．７３％）、５１ｂｐ（１１．１６％）、７０ｂｐ（ １５． ３２％）,与果蝇、蚤状溞的相同基因片段的序列含量基本相似。     

洪湖低湖田改造池塘河蟹养殖技术研究

在洪湖湖滨地区筛选了4种当地主要的河蟹(Eriocheir sinensis)养殖方式,对池塘与水源的环境因子进行了监测比较,分析各因子对河蟹收获参数的影响。结果显示:7—10月中,8月水温和氨氮含量最高,各池塘水体理化指标月平均值无显著差异;各池塘水草生物量逐月急剧下降,至10月底基本消失,其月平均值与成蟹体重、回捕率及特定生长率显著负相关。高温季节是防止河蟹氨氮中毒死亡的关键时期;水草调节水体营养、稳定底质、吸附水体悬浮质的功能对河蟹存活及生长有重要作用。当地优势池塘养殖方式为:放养规格约5 g/ind.,密度16875 ind./ha,水草移栽以伊乐藻为主,搭配其它种类,覆盖率约为60%,动物性饵料投喂比例约为66%,螺蛳投喂量约3638 kg/ha。     

虫草培养基残余体在中华绒螯蟹饲料中的适宜添加量

在基础饲料中分别添加0.25%、0.5%、1.0%、2.0%、4.0%、8.0%、16.0%的虫草培养基残余体(简称虫草基),饲喂规格为(4.0±1.0)g的中华绒螯蟹(Eriocheir sinensis)75 d,通过测定其生长及非特异性免疫指标,以确定虫草基的适宜添加量。结果显示:与基础饲料组比较,0.5%和1.0%添加组平均增重率(WGR)、肥满度(CF)、成活率(SR)显著提高(P<0.05),饲料系数(FC)显著降低(P<0.05);除肥满度(CF)外,4.0%、8.0%、16.0%添加组的生长指标有所下降。低剂量虫草基添加组能提高中华绒螯蟹血淋巴及肝胰腺超氧化物歧化酶(SOD)、溶菌酶(LSZ)、酸性磷酸酶(ACP)、碱性磷酸酶(ALP)活性,其中,0.5%与1.0%添加组的SOD、LSZ、ACP和ALP水平均达显著水平(P<0.05);但当虫草基添加量超过2.0%时,中华绒螯蟹的非特异性免疫力逐渐下降。嗜水气单胞菌(Aeromonas hydrophila AHJ-1)攻毒试验证实,试验组的免疫保护率明显提高,0.5%添加组免疫保护率达100%。综上所述,虫草基作为中华绒螯蟹饲料添加剂的适宜添加量为0.5%～1.0%。     

长江中游三个湖泊中华绒螯蟹生长变化研究

选择长江中游的牛山湖、武湖和东汤逊湖三个典型的河蟹养殖湖区作为研究地点,从2009年2月起逐月调查湖泊放流河蟹体重和壳宽的生长变化,并同时记录投喂管理情况和收集试验湖区的水化特征及底栖生物资源状况。调查发现,河蟹在4、5、6月特定生长率最大,此时为河蟹生长的高峰期。在三个湖区间,生长有显著性差异,在收获季节,武湖河蟹的平均壳宽和体重均显著性大于另外两个湖区(P<0.05),牛山湖和东汤逊湖之间则无显著性的差异(P>0.05);这种情形可能主要与为湖泊的天然饵料资源的种类和丰度有关。     

Effect of feeding scheme and prey density on survival and development of Chinese mitten crab Eriocheir sinensis zoea larvae

The effect of feeding scheme and prey density on survival and development of Eriocheir sinensis zoea larvae was studied in three experiments. Different combinations and densities of rotifers (Brachionus rotundiformis) and newly hatched Artemia nauplii were fed to zoea larvae. Average survival at each stage, larval development (larval stage index, LSI), duration of zoeal stage and individual megalopa dry weight were compared among treatments. This study revealed that, under the experimental conditions, rotifers should be replaced with Artemia between the zoea 3 (Z3) and the zoea 4 (Z4) stage. The optimal rotifer feeding densities for zoea 1 (Z1) and zoea 2 (Z2) were 15 and 20 mL^?1 respectively, while the optimal Artemia feeding density for Z3, Z4 and zoea 5 (Z5) was 3, 5 and 8 mL^?1 respectively. Further trials in production scale are recommended.     

中华绒螯蟹感染“颤抖病”病毒后血清免疫因子的变化

对中华绒螯蟹进行颤抖病病毒悬液和PBS(对照)注射感染,研究其血清免疫因子抗菌活性(Ua)、溶菌活性(UL)、凝集活性(HA)、酚氧化酶(PO)、过氧化酶(POD)、超氧化物歧化酶(S0D)、酸性磷酸酶(ACP)、碱性磷酸酶(AKP)活性的变化,并对实验的结果进行统计学分析。结果表明,在中华绒螯蟹人工感染颤抖病病毒后,蟹体内的免疫因子发生了明显的变化,血清中Ua、UL活性先是降低,然后上升至高于正常值水平。P0活性先上升然后下降至低于正常值水平,POD、HA、SOD活性逐渐升高至显著水平;ACP及AKP活性则显著下降。     

Effects of dietary protein level on growth,feed utilization and digestive enzyme activity of the Chinese mitten crab,Eriocheir sinensis

A feeding trial was conducted using isoenergetic practical diets to evaluate the effects of the dietary protein level on growth performance, feed utilization and digestive enzyme activity of the Chinese mitten crab, Eriocheir sinensis. Four experimental diets were formulated containing 250, 300, 350 and 400 g kg^?1 protein and 16 kJ g^?1 gross energy. Each diet was randomly assigned to triplicate groups of juvenile crab with mean initial body weight 3.56 ± 0.16 g and mean shell width 15.31 ± 0.06 mm. Juvenile crab were reared in indoor flow‐through system consisting of 12 plastic tanks (1.0 m × 0.6 m × 0.5 m) and fed diets twice daily at 6–8% of body weight for 12 weeks. Performance was judged on the basis of growth (specific growth rate of weight, SGR_G; specific growth rate of shell width, SGR_SW), feed conversion ratio (FCR) and protein efficiency ratio (PER). A decreased FCR was observed with increasing dietary protein levels. Both SGR_G and SGR_SW significantly increased with increasing dietary protein levels up to 350 g kg^?1, whereas there were no significant differences for protein levels from 350–400 g kg^?1. Application of broken line regression analysis to SGR_G provided an estimate of 347.8 g kg^?1 dietary protein for maximal growth. The highest PER was observed in crab fed the diet containing 350 g kg^?1 protein (P < 0.05). The percent survival was not affected (P > 0.05) by the different dietary treatments. No significant differences were observed in the apparent digestibility coefficients of crude lipid and dry matter among dietary treatments (P > 0.05). However, the apparent digestibility coefficients of crude protein and energy in crab fed different protein levels significantly increased with increasing dietary protein level (P < 0.05). Both amylase and protease activities in the intestine of E. sinensis were studied. The amylase activity decreased significantly (P < 0.05) with increased dietary protein level and protease activity increased. Regression analysis showed a negative effect of inclusion of dietary protein level on amylase activity (P < 0.05). However, protease activities were found to be positively correlated (P < 0.05) with dietary protein level. The protein content of the crab significantly increased with dietary protein levels up to 350 g kg^?1 (P < 0.05), but no significant differences (P > 0.05) were founded with protein levels higher than 350 g kg^?1.