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

A growth trial was conducted to estimate the optimum concentration of dietary Manganese (Mn) for grass carp (Ctenopharyngodon idella). Triplicate groups of grass carp (3.97 ± 0.05 g) were fed diets containing graded levels (4.0, 8.9, 13.8, 18.7, 23.6 and 33.3 mg kg^?1) of Mn for 8 weeks. Weight gain, specific growth rate and feed efficiency were linearly increased up to the 18.7 mg kg^?1 dietary Mn and then levelled off beyond this level. For body composition, lipid contents in whole body, muscle and liver decreased significantly with increasing dietary Mn level. Grass carp fed with dietary Mn levels higher than 19.7 mg kg^?1 significantly decreased condition factor. Whole body, vertebrae and scales mineralization were all affected significantly by dietary Mn levels. Mn contents in whole body, vertebrae and scales were linearly increased up to the 18.7 mg kg^?1 dietary Mn and then levelled off beyond this level. Contrarily, Ca and P contents seem to be inversely related to dietary Mn. However, dietary Mn levels had no significant effect on body Fe contents. Broken‐line analysis indicated that 20.6 mg kg^?1 dietary Mn was required for maximal tissue Mn storage, as well as satisfied for the optimal growth of juvenile grass carp.     

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.     

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

A growth trial was conducted to estimate the optimum concentration of dietary magnesium (Mg) for grass carp (Ctenopharyngodon idella). Triplicate groups of grass carp (5.56 ± 0.02 g) were fed diets containing graded levels (187, 331, 473, 637, 779 and 937 mg kg^?1) of Mg for 8 weeks. Weight gain, specific growth rate and feed efficiency were linearly increased up to 637 mg kg^?1 dietary Mg and then levelled off beyond this level. For body composition, dietary Mg levels higher than 473 mg kg^?1 significantly decreased the moisture content but increased the lipid content of whole body, muscle and liver. Dietary Mg levels higher than 473 mg kg^?1 significantly decreased the ash contents of vertebrae, scales and muscle. Mg contents in whole body, vertebrae, scales and plasma were increased up to 637 mg kg^?1 dietary Mg and then levelled off beyond this level. However, Ca and P contents seem to be inversely related to dietary Mg. Dietary Mg levels higher than 473 mg kg^?1 significantly decreased Zn and Fe contents in whole body and vertebrae. Broken‐line analysis indicated that 687 mg kg^?1 dietary Mg was required for maximal tissue Mg storage, as well as satisfied for the optimal growth.     

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.     

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.     

Quantifying the dietary potassium requirement of subadult grass carp (Ctenopharyngodon idellus)

A growth trial lasting for 12 weeks was conducted in 21 net cages to determine the dietary potassium (K) requirement of subadult grass carp (Ctenopharyngodon idellus) (Average weight: 331.3 g). Seven isonitrogenous and isoenergetic semi‐purified diets were compounded with different dietary K level. The specific growth rate (SGR) of fish was significantly (P < 0.05) improved by dietary K supplementation, SGR and the gill Na⁺‐K⁺ ATPase activity increased first and then decreased (P < 0.05) as dietary K level increased. The highest SGR and gill Na⁺‐K⁺ ATPase activity values were both observed at 6.38 g kg^?1 group. Dietary K level showed significant (P < 0.05) effect on serum superoxide dismutase (SOD) and glucose (GLU), the maximum values of SOD and GLU were in 8.42 and 6.38 g kg^?1 group, respectively. The body lipid content of the 6.38 g kg^?1 group was significantly (P < 0.05) lower than that of the control. However, the ash content in the 8.42 g kg^?1 group was significantly higher than those in the 1.21, 2.21, 4.41 and 6.38 g kg^?1 group. When dietary protein was 320 g kg^?1 and the waterborne potassium ranged from 6.86 to 9.10 mg L^?1, the dietary K requirement for subadult grass carp judged from SGR and gill Na⁺‐K⁺ ATPase activity is 5.38 and 7.41 g kg^?1 diet, respectively.     

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 magnesium requirements of juvenile grass carp,Ctenopharyngodon idella

To determine dietary magnesium (Mg) requirements of juvenile grass carp, Ctenopharyngodon idella, magnesium sulphate was added to the basal diet at 0, 150, 300, 600, 1200, 2400 mg Mg kg⁻¹ diet. Each diet was fed to three replicate groups of juvenile grass carp (initial weight: 7.69 ± 0.13 g) in a closed, recirculating rearing system for 76 days. No mortality or nutritional deficiency signs were observed except the growth depression in fish fed the Mg‐deficient diet. Growth performance and activities of serum superoxide dismutase (SOD), glutathione peroxidase (GPx) and lysozyme (LSZ) were highest (P <0.05) in fish fed the diet supplemented with 600 mg Mg kg⁻¹. The serum malondialdehyde (MDA) content was higher (P <0.05) in fish fed the diets supplemented with 0 and 150 mg Mg kg⁻¹ than that in fish fed the diets with ≥300 mg Mg kg⁻¹. Mg concentrations both in whole‐body and vertebrae increased with the increase in dietary Mg level up to 300 mg kg⁻¹, whereupon the response reached a plateau. Analysis by second‐order polynomial regression of weight gain, by broken‐line regression of vertebrae Mg concentration and by linear regression of whole‐body Mg retention of fish indicated that the adequate dietary Mg concentration for juvenile grass carp was 713.5, 627.7 and 469.8 mg kg⁻¹ diet, respectively.     

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.     

Total aromatic amino acid requirement of juvenile grass carp (Ctenopharyngodon idella)

This experiment was conducted to investigate total aromatic amino acid requirement of juvenile grass carp Ctenopharyngodon idella. Six isonitrogenous and isoenergetic semipurified diets containing casein and gelatin with graded level of phenylalanine (7.8, 11.1, 14.4, 17.6, 21.7, 24.9 g kg^?1 DM) were formulated. Each diet was randomly assigned to triplicate group of 30 fish (3.58 ± 0.002 g, mean ± SEM) each tank for 8 weeks. The highest weight gain (WG, %), final body weight (g) and specific growth rate were recorded when phenylalanine level was 17.6 g kg^?1 of the diet. Fish muscle protein content, protein efficiency ratio (PER), feed conversion ratio and alanine aminotransferase were significantly affected by dietary phenylalanine level. The polynomial regression calculated using WG and PER indicated that the optimal dietary total aromatic amino acid (phenylalanine + tyrosine) requirement for juvenile grass carp was 24.4 g kg^?1 of the diet, corresponding to 65.9 g kg^?1 of dietary protein.     

草鱼HSP70基因cDNA部分序列克隆及其组织表达差异

根据鲤热休克蛋白70(Heat shock protein,HSP70)序列(AY120894)设计并合成一对引物,以草鱼(Cteno-pharyngodon idella)肝胰脏组织总RNA为模板,RT-PCR扩增获得草鱼HSP70基因cDNA部分序列,并进行了组织表达差异性研究。结果显示:所获为序列为480 bp,获得GeneBank登陆号为FJ483832。序列测序结果显示,HSP70扩增序列与鲤、斑马鱼、鲋的同源性为:93%、91%、93%。另外,所获序列HSP70在草鱼脂肪、肌肉、肠、脑、粘液、性腺、鳔、肝胰脏、心脏、脾脏、鳃、鳍12个组织的表达存在差异,HSP70在草鱼这12个组织中均检测到表达,其中在鳍中表达最高,极显著高于其他组织(P<0.01);在鳔中表达次之,且与脑、心脏、性腺中表达差异不显著;在粘液中表达最低。     

不同脂肪源饲料对草鱼稚鱼生长的影响

不同脂肪源饲料对草鱼稚鱼生长的影响刘玮，徐萍，任本根，龚纲明（江西省科学院生物资源研究所，南昌３３００２９）关键词草鱼，鱼饲料，必采脂肪酸ＥＦＦＥＣＴＳＯＦＤＩＥＴＳＣＯＮＴＡＩＮＩＮＧＤＩＦＦＥＲＥＮＴＬＩＰＩＤＳＯＮＧＲＯＷＴＨＯＦＪＵＶＥＮＩＬ...     

长江草鱼群体MHC Class ⅡB的多态性和进化分析

克隆及测序草鱼(Ctenopharyngodon idella)长江3个群体的主要组织相容性复合体(MHC)Class II B基因编码β1和β2区的第2和第3个外显子及两个外显子之间的内含子,分析了草鱼MHC的进化模式和种群遗传结构。结果显示:实验共定义了34个等位基因,每条序列包括长为130~136 bp的第2个外显子,长为218 bp的第3个外显子以及长81~371 bp的内含子。序列分析揭示,第2个外显子有106个核苷酸变异位点(78%)和40个氨基酸变异位点(88%),而第3个外显子有100个核苷酸变异位点(45%)和41个氨基酸变异位点(56%),β1变异要大于β2区。用β1和β2区序列分别构建的邻接(NJ)系统树均显示5个具有高支持率的谱系,结合序列变异特点和内含子长度,推测草鱼至少存在5个MHC Class II座位。分别计算β1的肽结合位点(PBR)、非肽结合位点及β2的非同义替换率(dN)和同义替换率(dS),PBR的dN/dS为2.03(P<0.05),非肽结合位点和β2则小于1,表明草鱼MHC受到歧化选择作用。根据等位基因在群体中的分布频率作分子方差分析(AMOVA),得出FST为0.37%,提示长江草鱼MHC没有遗传分化。     

草鱼C1qC基因的克隆及表达分析

为了研究C1qC基因在草鱼(Ctenopharyngodon idella)免疫过程中所起的作用,利用RT-PCR和RACE方法克隆获得了C1qC基因cDNA全长序列,经序列分析表明,所克隆的C1qC cDNA全长为916 bp,包括开放阅读框(open reading frame,ORF)735 bp,5′端非编码区(untranslated region,UTR)89 bp和3′端非编码区(UTR)92 bp。735 bp的ORF共编码244个氨基酸,相对分子量为26 162.5 U。同源性分析表明,草鱼与斑马鱼(Danio rerio)的相似度最高,达到71%。经草鱼呼肠孤病毒(grass carp reovirus,GCRV)诱导后,草鱼C1qC基因在鳃、皮肤、肌肉、肝、中肾、心脏、头肾等组织中的mRNA表达水平均显著上调。在草鱼胚胎发育的各个阶段都能检测到C1qC mRNA的表达,说明该基因可能在草鱼胚胎和鱼苗的免疫反应和早期发育中起重要作用。本研究将为今后在草鱼免疫功能方面深入研究C1qC基因提供基础资料。     

Dietary potassium requirement of fingerling Indian major carp,Labeo rohita (Hamilton), based on growth parameters,gill Na+‐K+ ATPase activity,tissue mineralization and antioxidant activities

To quantify the dietary potassium requirement of fingerling Labeo rohita (6.2 ± 0.12 cm; 1.98 ± 0.06 g), seven purified experimental diets (350 g/kg crude protein and 16.72 kJ/g gross energy) with graded levels of potassium (0.32, 1.35, 2.41, 3.46, 6.48, 9.47 and 12.39 g/kg diet) were fed to triplicate groups of fishes at 08:00, 12:00 and 16:00 hr to apparent satiation for 8 weeks. Live weight gain (LWG; 671.46%), specific growth rate (3.65%/day), protein efficiency ratio (2.16), protein gain (PG; 2.41 g/fish) and feed conversion ratio (1.32) were found to be best in fish fed diet containing 3.46 g/kg potassium. Gill Na⁺‐K⁺ ATPase activity was also highest in fish fed diet with 3.46 g/kg potassium. Potassium content of whole‐body, vertebrae and scales increased significantly with the increase in dietary potassium level up to 6.48 g/kg. Significant changes were also noted in serum malondialdehyde content, superoxide dismutase, catalase, glutathione peroxidase and alkaline phosphatase activity. Based on the maximum live weight and protein gain observed in the present study, the inclusion of 3.55 g/kg potassium is recommended for developing potassium‐balanced commercial feeds for intensive culture of fingerling L. rohita.     

投饲蚕豆对草鱼生长和肉质影响的初步研究

分别以配合饲料(对照组)、蚕豆(蚕豆组)饲喂平均体重为496.5 g的草鱼(Ctenopharyngodon idella)66d,考察蚕豆对草鱼生长和肉质的影响。结果表明:配合饲料组和蚕豆组增重率为82.76%,36.96%;饵料系数为2.17,4.45;肥满度为13.09,7.72;粗脂肪含量为0.89%,1.43%;肌肉失水率为27.14%,15.87%;肌原纤维长度为28.32μm,33.31μm;羟脯氨酸0.12mg/g,0.16mg/g。在生长方面,与配合饲料组相比,蚕豆组草鱼的增重率、肥满度极显著降低,饲料系数极显著增加(P<0.01);在肌肉品质方面,失水率极显著降低;粗脂肪含量、肌原纤维长度、羟脯氨酸含量极显著提高(P<0.01)。结果表明,投喂蚕豆对草鱼肌肉具有特殊的改变作用。     

饲料中添加谷胱甘肽对草鱼组织中谷胱甘肽沉积和抗氧化能力的影响

在基础饲料中分别添加还原型谷胱甘肽(GSH)至0 mg/kg、100 mg/kg、200 mg/kg、300 mg/kg、400 mg/kg和500 mg/kg,分别投喂草鱼(Ctenopharyngodon idella)幼鱼8周,草鱼幼鱼体质量(4.09±0.01)g,观察GSH在草鱼组织中沉积以及对草鱼抗氧化功能的影响.结果表明,饲料中添加外源GSH对草鱼生长影响不显著(P＞0.05),实验组肌肉中GSH含量显著高于对照组(P＜0.05),肝脏中GSH含量在GSH添加水平为200 mg/kg时显著高于对照组(P＜0.05),肝脏和肌肉中丙二醛(MDA)在GSH添加水平为300 mg/kg组达到最低,显著低于对照组(P＜0.05).添加GSH对血清中GSH和MDA影响不显著(P＞0.05).草鱼肝脏中谷胱甘肽还原酶(GR)活力在400 mg/kg组显著高于对照组(P＜0.05),肌肉中GR活力有增高趋势但差异不显著(P＞0.05);肝脏和肌肉中γ-谷氨酰转移酶(γ-GT)均高于对照组,分别在300 mg/kg组和200 mg/kg组达到显著水平(P＜0.05);肝脏中谷胱甘肽过氧化物酶(GSH-PX)活力、总抗氧化能力(T-AOC)和超氧化物歧化酶(SOD)能力不同程度升高,均在200 mg/kg组达到最高,显著高于对照组(P＜0.05);血清GSH-PX活力和T-AOC有增高趋势但与对照组差异不显著(P＞0.05);血清和肝脏中活性氧(ROS)含量分别在400 mg/kg和300 mg/kg组达到最低,显著低于对照组(P＜0.05).结果提示,饲料中添加GSH能够促进草鱼肝脏和肌肉中GSH的沉积,提高肝脏及肌肉中GR和γ-GT活力,以及肝脏中GSH-PX和SOD活力与总抗氧化能力,减少肝脏中MDA含量,降低肝脏及血清中ROS含量,因此GSH在水产饲料中具有广泛的应用前景.     

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.     

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.