Qualitative and quantitative strategies of thermal adaptation of grass carp (Ctenopharyngodon idella) cytoplasmic malate dehydrogenases

Malate dehydrogenase isozymes of grass carp,Ctenopharyngodon idella, were identified by mitochondrial preparation and thermal denaturation. The structural and kinetic characteristics of chromatographically separated thermostable and thermolabile cMDHs were different in (1) half-life at 42°C, 10 min and 24 h, respectively, (2) optimal substrate, oxaloacetate and malate, concentrations, and (3) the apparent Michaelis-Menten constants of NADH and oxaloacetate.Total MDH activity in white muscle of 11°C-acclimated fish was about twice that of the 30°C-acclimated group. In addition, the ratio of the thermostable to thermolabile cMDH activity in white muscle of 30°C-acclimated fish was significantly higher than that of 11°C-acclimated fish. These results suggest that temperature acclimation can induce temperature compensation in MDH activity and differential expression of thermostable and thermolabile cMDH isozymes in freshwater fish.     

The influence of feeding rate on growth, feed efficiency and body composition of juvenile grass carp (Ctenopharyngodon idella)

An 8 weeks growth study was conducted to estimate the optimal feeding rate for juvenile grass carp (3.08±0.03 g, mean ± SD). Fish were fed with a casein purified diet (360 g protein, 56 g lipid and 3000 kcal total energy/kg dry diet) at six feeding rates: 1.0, 1.5, 2.0, 2.5, 3.0, 3.5% body weight per day (BW d⁻¹). Each feeding rate was randomly assigned to three tanks of fish with 30 fish per tank (50W × 50H × 100L, cm). Fish were maintained in recirculating systems at a water temperature of 24.97±2.23 °C and were fed four times per day. After 2 weeks, fish fed on 3.5% BW d⁻¹ could not finish the diet and this treatment was cut-off. Analysis of variance showed that growth performance was significantly (p<0.05) affected by different feeding rates. The nutrient compositions of whole body, muscle and liver were also significantly different among treatments. The body weight gain (WG), specific growth rate (SGR), feed efficiency (FE), protein efficiency ratio (PER), apparent digestibility coefficiency (ADC), retention of protein (PR), mesenteric fat index, body moisture and protein content were significantly (p<0.05) affected by feeding rate. The WG, SGR and digestion rate were highest at 2% BW d⁻¹, although the FE and PER decreased with increasing feeding rate. Broken line analysis on specific growth rate indicated that the optimum feeding rate of juvenile grass carp is 1.97% body weight day⁻¹.     

Dietary calcium requirement and effects on growth and tissue calcium content of juvenile grass carp (Ctenopharyngodon idella)

A growth trial was conducted to estimate the optimum concentration of dietary calcium (Ca) for grass carp (Ctenopharyngodon idella). Triplicate groups of grass carp (4.52 ± 0.02 g) were fed diets containing graded levels (2.75, 4.51, 6.24, 7.99, 9.66 and 11.5 g kg^?1) of Ca for 8 weeks. Weight gain, feed efficiency and protein efficiency ratio were linearly increased up to the 7.99 g kg^?1 dietary Ca and then maintained stable beyond this level (P < 0.05). Dietary Ca levels higher than 7.99 g kg^?1 significantly increased the ash contents of whole body, vertebrae and scales. Ca contents in whole body, vertebrae and scales were linearly increased up to the 7.99 g kg^?1 dietary Ca and then maintained stable beyond this level (P < 0.05). In contrast, dietary Ca levels higher than 9.66 g kg^?1 significantly decreased Mg contents in whole body, vertebrae and scales. Dietary Ca levels higher than 7.99 g kg^?1 significantly increased plasma alkaline phosphatase activity. However, plasma Ca, P and Mg contents were not significantly affected by dietary Ca supplements (P > 0.05). Polynomial regression analysis indicated that 10.4 g kg^?1 dietary Ca was required for maximal tissue storage and mineralization as well as optimal growth.     

Grass carp (Ctenopharyngodon idella) grazing on duckweed (Spirodela polyrhiza)

The aims of this experiment were (1) toquantify the ability of grass carp to processduckweed and (2) to assess indirect changes inwater chemistry and phytoplankton community,caused by grass carp feeding. Yearling grass carp sized 126 ± 7.7 mm (TL) and19.6 g in weight were kept in 9 laminate tanksof 1 m³ for 14 days. Two stockingdensities (2 and 6 fish per m³) anda control without fish were used. Standard growthrate (SGR) of grass carp fed exclusively onduckweed was 0.70% body weight (BW) d^–1and food conversion ratio (FCR) reached 2.0(average water temperature =21.1 ± 3.8 °C). Daily food intakewas 0.2 g of duckweed dry weight (DW), i.e.,1% of average BW of grass carp. SGR ofduckweed growing in 20 × 20 cm floatingenclosures, differed significantly[F(6,2) = 417.9; p = 0.002] between the twostocking densities of grass carp and thecontrol tanks (without fish). Mean SGR ofduckweed was 0.02 g g^–1 day^–1 and thehighest SGR was recorded in the control tanks.Both decrease in NH₄-N and increase inNO₂-N concentrations differedsignificantly between the treatments[F(2,2) = 45.3; p = 0.02 and F(2,2) = 19.2; p = 0.04 respectively]. Changes in other nitrogenand phosphorus components (NO₃-N, TN, TPand PO₄-P) caused by stocking of grasscarp were not significant. Biomass ofphytoplankton, dominated by filamentous algaeand blue-greens, increased proportionately tostocking density of grass carp. Althoughduckweed has a large potential for nutrientremoval, the most common pathway for thenutrients released through grass carp grazingif duckweed cover is loose is theirincorporation into phytoplankton biomass.     

Dietary available phosphorus requirement of juvenile grass carp (Ctenopharyngodon idella)

A growth trial was conducted to estimate the optimum concentration of dietary available phosphorus (P) for grass carp (Ctenopharyngodon idella). Triplicate groups of grass carp (5.59 ± 0.02 g) were fed diets containing graded levels (2.36, 4.27, 6.31, 8.36, 10.4 and 14.8 g kg^?1) of available P for 8 weeks. Grass carp fed with the P‐supplemented diets had significantly higher specific growth rate, weight gain, protein efficiency ratio and feed efficiency than fish fed with the basal diet. In whole‐body composition, protein content increased, while lipid content decreased with the increase in P level in diet (P < 0.05). Fish fed with the P‐supplemented diets had significantly higher whole body, vertebrae and scales mineralization (P < 0.05), but Ca/P ratios were not influenced. The blood chemistry analysis showed that dietary available P had distinct effects on P, Ca and Mg contents, as well as on the contents of triacylglycerol and total cholesterol. Broken‐line analysis indicated that 8.49 g kg^?1 dietary available P was required for maximal tissue storage and mineralization as well as optimal growth.     

Dietary potassium requirement of juvenile grass carp (Ctenopharyngodon idella Val.) based on growth and tissue potassium content

A growth trial was conducted to estimate the optimum concentration of dietary potassium (K) for grass carp (Ctenopharyngodon idella). Triplicate groups of grass carp (3.96 ± 0.06 g) were fed diets containing graded levels (0.87, 2.90, 5.37, 7.54, 9.87 and 12.4 g kg^?1) of K for 8 weeks. Final body weight, weight gain and feed efficiency and gill Na⁺‐K⁺ ATPase activity were highest in fish fed with 9.87 g kg^?1 dietary K and lowest in fish fed the basal diet (P < 0.05). The K contents in whole body and muscle were linearly increased up to the 9.87 g kg^?1 dietary K and then levelled off beyond this level, whereas in scales and vertebrae up to the 7.54 g kg^?1 dietary K (P < 0.05). However, dietary K levels had no significant effect on ash, Ca, P and Mg contents in whole body, scales, vertebrae or muscle. Analysis using polynomial regression of weight gain and gill Na⁺‐K⁺ ATPase activity and using the broken‐line regression of whole body K concentrations indicated that the adequate dietary K concentration for grass carp is about 9.45–9.99 g kg^?1 diet.     

Dietary arginine requirement of juvenile grass carp Ctenopharyngodon idella (Valenciennes in Cuvier & Valenciennes, 1844)

This experiment was conducted to estimate the optimum requirement of arginine for juvenile grass carp Ctenopharyngodon idella. Six isonitrogenous (38%) and isoenergetics (16 MJ kg^?1) semi‐purified diets containing casein and gelatine with graded level of arginine (0.93, 1.20, 1.51, 1.84, 2.10 and 2.41 g 100 g^?1 DM) were formulated. Each diet was randomly assigned to triplicate groups of 25 fish each tank (initial weight: 3.84 ± 0.01) for 10 weeks. The highest weight gain (WG, %) was recorded when arginine level was 2.10% of the diet. Dietary arginine level higher than 1.84% significantly increased the protein contents of whole body. Whole body amino acid composition of juvenile grass carp was not significantly affected by the dietary arginine level. Plasma‐free arginine level was increased linearly with increasing of arginine level in the diets, and the plasma‐free ornithine level was significantly higher when the dietary arginine level was 2.41% compared with other groups. Quadratic model analysis of SGR data indicated that the minimum recommended dietary arginine requirement for grass carp was 2.17% of the diet, corresponding to 5.71% of dietary protein.     

Effects of dietary phosphorus and starch levels on growth performance,body composition and nutrient utilization of grass carp (Ctenopharyngodon idella Val.)

Six iso‐nitrogenous (410 g kg^?1) diets with three levels of total phosphorus (P4, P10 and P18 g kg^?1) and two levels of starch (S200 and S350 g kg^?1) were fed to triplicate groups of 30 fish to evaluate whether the high level of dietary phosphorus could improve the utilization of starch. Over 8‐week‐growth trial, best weight gain (WG) and specific growth rate (SGR) (P < 0.05) were observed in fish fed the P10/S200 and P18/S200 diets. WG and SGR significantly decreased as starch levels increased whereas for P4, while lipid contents of liver and whole body, hepatosomatic index and intraperitoneal fat ratio (IPF) significantly increased. These results suggested that high dietary starch will depress the growth performance and cause lipid accumulation. Within both starch levels, fish fed diet with P4 tended to produce lower (P < 0.05) WG and SGR, and had higher (P < 0.05) values of IPF. The whole body lipid, ash, calcium, phosphorus and iron contents were significantly affected by dietary phosphorus levels. Supplied phosphorus could improve the growth and decrease the whole body lipid, but there is no more effect after the phosphorus requirement was met at 10 g kg^?1.     

Dietary zinc requirement of juvenile grass carp (Ctenopharyngodon idella) based on growth and mineralization

A growth trial was conducted to estimate the optimum requirement of dietary zinc (Zn) for grass carp (Ctenopharyngodon idella). Triplicate groups of grass carp (3.97 ± 0.05 g) were fed diets containing graded levels (13, 25, 34, 53, 89 and 135 mg kg^?1) of Zn for 8 weeks. Grass carp fed with dietary Zn levels higher than 34 mg kg^?1 significantly increased final body weight, weight gain and specific growth rate (P < 0.05). For body composition, fish fed with dietary Zn levels higher than 53 mg kg^?1 significantly decreased the moisture contents but increased the lipid contents of whole body and liver. Whole body, scales, vertebrae and liver mineralization were all affected significantly (P < 0.05) by dietary Zn levels. Zn contents in whole body, scales, vertebrae and plasma were linearly increased up to the 53 mg kg^?1 dietary Zn and then remained stable beyond this level. Grass carp fed with dietary Zn levels higher than 53 mg kg^?1 significantly increased triacyglyceride and total cholesterol contents and plasma alkaline phosphatase activity in plasma (P < 0.05). Broken‐line analysis indicated that 55.1 mg kg^?1 dietary Zn was required for maximal tissue storage and mineralization as well as optimal growth of grass carp.     

草鱼呼肠孤病毒纤维蛋白VP56与草鱼GRP78蛋白互作诱导内质网应激

为了阐明草鱼呼肠孤病毒(GCRV)的感染机制,深入探究VP56 (Ⅱ型和Ⅲ型GCRV特有的纤维蛋白)与宿主细胞蛋白的相互作用。实验通过免疫共沉淀(co-IP)发现,VP56与细胞内定位于内质网的分子伴侣蛋白葡萄糖调节蛋白78 (GRP78)发生相互作用。而后,VP56与GRP78的互作通过基于远红外红色荧光蛋白mNeptune的双分子荧光互补系统(BiFC)得到验证。在VP56稳定表达的草鱼肾细胞系(CIK)中,相较于CIK细胞,内质网则形态发生巨大变化,产生肿胀、扩张、脱粒等现象,说明VP56激活内质网应激。通过对内质网应激下游转录因子的mRNA水平检测,发现VP56激活活化转录因子6 (ATF6)介导的信号转导途径,激活非折叠蛋白反应。研究表明,GCRV-Ⅱ感染过程中破坏了细胞稳态、激活内质网应激。本研究为GCRV感染机制和抗GCRV病毒研究提供了新思路,揭示了一种新的病毒逃逸机制,有助于淡水养殖产业中草鱼出血病的防控。     

地衣芽孢杆菌De在优质草鱼养殖中的应用研究

运用综合对比分析法探讨了地衣芽孢杆菌Bacillus licheniformis De在优质草鱼（Ctenopharyngodon idellus）养殖中的应用效果,其评价指标分别为成活率、水体pH、透明度、溶解氧及水中氨氮、硝酸盐浓度等。结果表明,施用地衣芽孢杆菌De可在一定程度上使水体环境和养殖生产性能得到优化,提高养殖草鱼的成活率,显著降低水体透明度及水中氨氮、硝酸盐含量（P〈0．05）,使水体pH、溶解氧有利于草鱼的生长。其中施菌组较对照组的成活率、水体pH、溶解氧分别提高了3．2％、3．9％、25．5％,而水体透明度、氨氮及亚硝氮浓度则分别降低了38．5％、74．6％、69．3％。     

Partial characterization and activity distribution of proteases along the intestine of grass carp, Ctenopharyngodon idella (Val.)

A series of studies based on biochemical assays and electrophoretical observations has been conducted in order to investigate activity distributions and partially characterize various types of proteinases in the digestive tract of grass carp, Ctenopharyngodon idella (Val.). The casein digestion assays revealed that the presence of acidic proteinase had the highest activity at pH 2.5–3.0 and the alkaline proteinases at pH 10.0. The acidic proteinase activity distribution was found to decrease gradually from the oesophagus to the anus. Pepstatin A and EDTA inhibited the acidic proteinases activity. The SDS‐substrate‐PAGE showed that crude extraction of grass carp intestine contained an acidic proteinase active component with molecular mass of 28.5 ku. The substrate‐PAGE at neutral pH condition showed the presence of two acidic proteinase active components. The activity distribution of alkaline proteinase was found to slightly fluctuate along the intestine. And the whole intestine had very high activity. The inhibition assays and substrate specificity assays showed that trypsin was the main active component of the alkaline proteinases. The SDS‐substrate‐PAGE further showed that the crude extraction of grass carp intestine had four types of alkaline proteinase with molecular mass of 26.4, 30.8, 43.0 and 105.0 ku respectively. They were characterized to be trypsin (26.4, 30.8 and 43.0 ku) and un‐serine proteinase (105.0 ku) respectively. No chymotrypsin was detected.     

Dietary quercetin improved the growth,antioxidation, and flesh quality of grass carp (Ctenopharyngodon idella)

To investigate the effects of dietary quercetin on growth, antioxidation, and flesh quality of grass carp, Ctenopharyngodon idella, six diets were prepared with quercetin inclusion rates of 0 (control diet), 0.1, 0.2, 0.4, 0.6, and 0.8 g/kg. Grass carp with a body weight of 13.3 ± 0.1 g were fed with one of the six diets for 60 days. The weight gain (WG) showed a quadratic relationship with dietary quercetin levels; the supplementation of 0.4 g/kg quercetin significantly improved WG (+4.73%) and decreased feed conversion ratio (?0.06) (p < .05) when compared to those of the control group. The intestinal fat ratio was reduced by the addition of 0.2 or 0.4 g/kg of quercetin (p < .05), and serum activities of alkaline phosphatase and superoxide dismutase were increased by the addition of 0.4 and 0.6 g/kg of quercetin (p < .05). The inclusion of 0.2–0.6 g/kg of quercetin increased the contents of delicious amino acids and decreased the cooking loss of flesh (p < .05). Flesh collagen content was increased by the addition of 0.4–0.8 g/kg of quercetin (p < .05). In conclusion, dietary quercetin could improve the growth and enhance the antioxidation and flesh quality of grass carp, with the recommended supplemental level of quercetin was 0.37 g/kg.     

饲喂枯草芽孢对草鱼粪便、养殖水体中芽孢数量和水质的影响

分别在饲料中添加不同含量的枯草(Bacillus subtilis)芽孢(A组实测芽孢数量为2.6×107 cfu/g,B组为4.4×106 cfu/g,空白组为0),饲喂静水水族箱中100 g左右的草鱼(Ctenopharyngodon idellus)10 d,测定饲喂枯草芽孢后草鱼粪便、养殖水体中芽孢数量和水质的变化。结果表明:(1)9 d内,水体中芽孢数量A组和B组在前6 d时不断增加,在6 d时达到稳定,其中A组和B组稳定时的含量分别为2.22×106 cfu/g和3.98×105 cfu/g。(2)10 d内,草鱼粪便干物质中的芽孢数量与饲料中的枯草芽孢含量相比,A组和B组粪便中的枯草芽孢损失率分别为89.76%～90.71%和83.83%～86.84%。(3)草鱼饲喂枯草芽孢后,A组和B组粪便中排出的枯草芽孢对水体中亚硝酸盐和氨氮有降低的趋势,但各组之间差异不显著(P>0.05),对化学需氧量(COD)没有影响。     

Use of probiotics Bacillus coagulans,Rhodopseudomonas palustris and Lactobacillus acidophilus as growth promoters in grass carp (Ctenopharyngodon idella) fingerlings

The present research evaluated the effects of three probiotics on the growth performance and intestinal digestive enzyme activity in fingerlings of grass carp, Ctenopharyngodon idella. Three treatments (T‐1, T‐2 and T‐3) were fed with diets containing different viable bacteria with a final concentration 10⁶ CFU g^?1 feed (Bacillus coagulans, Rhodopseudomonas palustris and Lactobacillus acidophilus, respectively) and the control was fed with basal diet without probiotics. All the diets supplemented with probiotics resulted in better (P < 0.05) final weight, daily weight gain and relative gain rate than control but no significant differences were observed among the treatment groups. The highest protease activity was observed in T‐1. However, T‐2 and T‐3 showed no difference (P > 0.05) in protease and cellulase activities compared with the control. The protease activity was higher in the foreintestine than in the hindintestine (P < 0.05), and the reverse was observed in cellulase activity. As for amylase activity, there was no difference between foreintestine and hindintestine. In conclusion, the three selected probiotics increased the growth performance of grass carp fingerlings. Furthermore, different digestive enzyme activity was observed in different intestine segment.     

单剂量口灌阿维菌素在草鱼体内的药动学及残留研究

按0.1 mg/kg的剂量给草鱼(Ctenopharyngodon idellus)灌服阿维菌素,用高效液相色谱法检测用药后不同时间的血浆、肌肉和肝脏中的药物浓度,然后用3P97药代动力学软件处理药时数据,对药物在草鱼体内药动学及组织残留进行研究.结果表明:单剂量口灌阿维菌素在草鱼血浆中主要药动学参数为:AUC 1 6...     

不同电学参数和流速下的拦鱼电栅对草鱼幼鱼的拦导效率

为了探究不同电学参数的拦鱼电栅在静水条件下对草鱼幼鱼[体长(10.22±2.01)cm、体质量(34.25±3.62)g]趋避行为的影响,实验构建了一种双排式的拦鱼电栅,在静水条件下采用3因素4水平正交实验设计,测试了脉冲电压、脉冲频率、脉冲宽度等脉冲直流电参数对拦鱼效果的影响;为了探究静水下拦鱼效果最佳的拦鱼电栅电学...     

Molecular cloning of fatty acid synthase from grass carp (Ctenopharyngodon idella) and the regulation of its expression by dietary fat level

The fatty acid synthase (FAS) gene was cloned from liver of grass carp (Ctenopharyngodon idella) by degenerate oligonucleotide primed PCR. The obtained cDNA fragment was 683 bp, which encoded 227 amino acids. Then, grass carps with initial body weight of (134.89 ± 12.12) g were fed diet supplemented with 0, 20, 40 g kg^?1 fat to investigate the impacts of dietary fat levels on growth, liver FAS enzyme activity and mRNA expression. After 8 weeks feeding, final body weights of the three groups were 344.11, 347.23 and 373.02 g. Compared with control group (0 g kg^?1), growth rate of 40 g kg^?1 fat group was increased by 14.03%, and feed conversion rate decreased by 11.32% (P < 0.05), liver FAS enzyme activity of 20, 40 g kg^?1 fat groups were reduced by 33%, 64% (P < 0.05), and FAS mRNA expression level reduced by 18%, 74% (P < 0.05), respectively. Results above showed that 40 g kg^?1 fat addition can significantly improve growth performance of grass carp. Liver FAS activity and mRNA expression tended to be inhibited by the increasing dietary fat level. Fat containing high levels of polyunsaturated fatty acids had strongly inhibitory effects on liver FAS activity and gene expression.     

池塘两种养殖方式下草鱼的营养差异

为了解传统池塘养殖和池塘内循环系统养殖草鱼的营养差异,对2种养殖方式草鱼的营养成分及营养品质进行了比较研究。结果显示,池塘养殖草鱼肌肉水分、粗脂肪含量、脏体比以及pH降低值和滴水损失高于循环养殖系统的草鱼,而肌肉粗蛋白含量和抗氧化能力低于循环养殖系统的草鱼。池塘养殖和循环系统养殖草鱼肌肉必需氨基酸构成比例均符合FAO/WHO的标准,必需氨基酸均是赖氨酸(Lys)含量最高;池塘养殖草鱼氨基酸总量(∑TAA)、鲜味氨基酸总量(∑DAA)和甜味氨基酸总量(∑SAA)低于循环养殖系统;根据氨基酸评分(AAS)和化学评分(CS),2种模式的第一限制性氨基酸均为缬氨酸(Val)。循环养殖系统草鱼肌肉∑n-3PUFA和∑n-6PUFA含量显著低于池塘养殖,但肌肉EPA+DHA含量是池塘养殖的1.5倍。研究表明,草鱼营养组成合理,池塘内循环系统养殖的草鱼在营养成分及鲜甜味方面优于池塘养殖。