低蛋白饲料对杂交黄颡鱼幼鱼生长性能、体组成、转氨酶活性和抗氧化能力的影响

本实验以杂交黄颡鱼(Pelteobagrus fulvidraco♀ ×Pelteobagrus vachelli♂)幼鱼为研究对象,探究低蛋白饲料中补充蛋氨酸和赖氨酸对杂交黄颡鱼的生长性能、体组成、转氨酶活性及抗氧化能力的影响.实验设计5组饲料,设置3个蛋白梯度(42％、37％和32％),分别记为对照组、LP1组和L...     

Human mitochondrial function during cardiac growth and development

Little information is presently available concerning mitochondrial respiratory and oxidative phosphorylation function in the normal human heart during growth and development. We investigated the levels of specific mitochondrial enzyme activities and content during cardiac growth and development from the early neonatal period (10-20 days) to adulthood (67 years). Biochemical analysis of enzyme specific activities and content and mitochondrial DNA (mtDNA) copy number was performed with left ventricular tissues derived from 30 control individuals. The levels of cytochrome c oxidase (COX) and complex V specific activity, mtDNA copy number and COX subunit II content remained unchanged in contrast to increased citrate synthase (CS) activity and content. The developmental increase in CS activity paralleled increasing CS polypeptide content, but was neither related to overall increases in mitochondrial number nor coordinately regulated with mitochondrial respiratory enzyme activities. Our findings of unchanged levels of cardiac mitochondrial respiratory enzyme activity during the progression from early childhood to older adult contrasts with the age-specific regulation found with CS, a Krebs cycle mitochondrial enzyme.     

Family origin and the response of threespine stickleback, Gasterosteus aculeatus, to thermal acclimation

To establish whether family origin affects the response of the threespine stickleback (Gasterosteus aculeatus) to thermal acclimation, we examined the rates of feeding, growth, and food conversion, relative tissue and organ masses and activities of a mitochondrial and a glycolytic enzyme in pectoral and axial muscle of individually housed fish from six families during acclimation to 8 degrees C and 23 degrees C. Feeding rates differed among families but were consistently higher in warm-acclimated than cold-acclimated fish. Growth rates differed among families. In four families growth was greater at 8 degrees C; these families generally had higher conversion efficiencies at 8 degrees C than 23 degrees C. For two families, growth was greater at 23 degrees C than 8 degrees C and conversion efficiencies did not differ between 8 degrees C and 23 degrees C. Relative tissue and organ masses (percent axial muscle, hepatosomatic, gut and kidney indices) differed with gender and among families (hepatosomatic, gut and kidney indices) but little with acclimation status. In all families and in both muscles, activities of the mitochondrial enzyme, citrate synthase (CS), were increased by cold acclimation. Axial muscle levels of the glycolytic enzyme, lactate dehydrogenase (LDH), were not affected by thermal acclimation or family origin, but were strongly correlated with the hepatosomatic index and axial muscle protein content. Pectoral muscle levels of LDH were affected by family origin which also influenced the response to thermal acclimation. Similar patterns were observed for specific activities and total muscle contents of these enzymes. Stickleback family origin influenced rates of feeding and growth and the thermal sensitivity of growth rates but not the compensatory increase in muscle CS levels with cold acclimation. The differing thermal sensitivities of growth could reflect distinct strategies for the timing of juvenile growth.     

Effect of dietary lysine on performance and expression of electron transport chain genes in the pectoralis major muscle of broilers

The aim of this study was to evaluate the effect of dietary lysine on performance, protein deposition and respiratory chain gene expression in male broilers. A total of 252 Cobb 500 broilers were distributed, in a completely randomized design, into four treatments with seven replicates of nine birds per experimental unit. Experimental treatments consisted of diets based on corn and soybean meal, with four levels of digestible lysine: 1.016%, 1.099%, 1.182% and 1.265%. The increase in the level of digestible lysine in the diet provided higher weight gains, feed efficiency and body protein deposition. Birds fed the lowest level of dietary lysine (1.016%) showed a lower expression of genes such as NADH dehydrogenase subunit I (ND1), cytochrome b (CYTB) and cytochrome c oxidase subunits I (COX I), II (COX II) and III (COX III), displaying the worst performance and body protein deposition. This demonstrates the relationship existing between the expression of the evaluated genes and the performance responses. In conclusion, results indicate that broilers fed diets with higher levels of digestible lysine have increased messenger RNA expression of some genes coded in the mitochondrial electron transport chain (ND1, CYTB, COX I, COX II and COX III). It may be stated that diets with proper levels of digestible lysine, within the ‘ideal protein’ concept, promote the expression of genes, which increases the mitochondrial energy, thereby fostering body protein deposition and the performance of broilers in the starter phase.     

梯棱羊肚菌AA1_2家族多铜氧化酶基因的异源表达与生化鉴定

典型的漆酶通常属于辅助活性酶第一家族第一亚族（auxiliary activity family 1 subfamily 1,简称AA1_1家族）,而AA1_2家族的多铜氧化酶通常拥有将二价铁氧化成三价铁的活性,部分AA1_2家族酶蛋白兼具漆酶活性。梯棱羊肚菌全基因组只有一个AA1_2家族酶基因,该基因编码的酶蛋白是否拥有漆酶功能尚未清楚。本研究主要从酶生化特性的角度,结合酶基因的表达规律,对该基因的功能进行初探。对该AA1_2家族基因在梯棱羊肚菌生长发育不同阶段的表达水平进行实时定量PCR检测;将该基因编码序列克隆到表达载体中在大肠杆菌中异源表达,层析获得纯化的酶蛋白,对酶蛋白的生化特性进行了鉴定。发现该AA1_2多铜氧化酶基因在外源营养袋和土壤中的营养菌丝里低表达,在菇原基和子实体中表达较活跃。异源表达获得纯化的酶蛋白分子量约64kDa,表现出亚铁氧化酶（EC 1.16.3.1）与漆酶（EC 1.10.3.2）双重活性。其亚铁氧化酶活性在pH 4最高,漆酶活性在pH 6最高。亚铁氧化酶活性与漆酶活性的最适温度均为30℃左右,在30℃温育16h后仍保留70%以上活性。亚铁氧化酶和漆酶活性受Mn ²⁺、Hg ²⁺和Pb ²⁺抑制。对蛋白质变性剂SDS、尿素的耐受性较强。本研究通过酶学证据证实了梯棱羊肚菌AA1_2家族多铜氧化酶基因编码的酶蛋白具有亚铁氧化酶-漆酶双重活性,系在子囊菌大型真菌中首次发现,为进一步研究铁元素代谢与漆酶活性在羊肚菌子实体形成与发育过程中的作用提供了启示。     

Hepatic glucokinase and glucose-6-phosphatase responses to dietary glucose and starch in gilthead sea bream (Sparus aurata) juveniles reared at two temperatures

The effects of carbohydrate sources/complexity and rearing temperature on hepatic glucokinase (GK) and glucose-6-phosphatase (G6Pase) activities and gene expression were studied in gilthead sea bream juveniles. Two isonitrogenous (50% crude protein) and isolipidic (19% crude lipids) diets were formulated to contain 20% waxy maize starch or 20% glucose. Triplicate groups of fish (63.5 g initial body weight) were fed each diet to near satiation during four weeks at 18 degrees C or 25 degrees C. Growth, feed intake, feed efficiency and protein efficiency ratio, were higher at the higher water temperature. At each water temperatures fish growth and feed efficiency were higher with the glucose diet. Plasma glucose levels were not influenced by water temperature but were higher in fish fed the glucose diet. Hepatosomatic index and liver glycogen were higher at the lower water temperature and within each water temperature in fish fed the glucose diet. No effect of water temperature on enzymes activities was observed, except for hexokinase and GK which were higher at 25 degrees C. Hepatic hexokinase and pyruvate kinase activities were not influenced by diet composition, whereas glucose-6-phosphate dehydrogenase activity was higher in fish fed the glucose diet. Higher GK activity was observed in fish fed the glucose diet. GK gene expression was higher at 25 degrees C in fish fed the waxy maize starch diet while in fish fed the glucose diet, no temperature effect on GK gene expression was observed. Hepatic G6Pase activities and gene expression were neither influenced by dietary carbohydrates nor water temperature. Overall, our data suggest that in gilthead sea bream juveniles hepatocytes dietary carbohydrate source and temperature affect more intensively GK, the enzyme responsible for the first step of glucose uptake, than G6Pase the enzyme involved in the last step of glucose hepatic release.     

Hepatic glucokinase and glucose-6-phosphatase responses to dietary glucose and starch in gilthead sea bream (Sparus aurata) juveniles reared at two temperatures.

The effects of carbohydrate sources/complexity and rearing temperature on hepatic glucokinase (GK) and glucose-6-phosphatase (G6Pase) activities and gene expression were studied in gilthead sea bream juveniles. Two isonitrogenous (50% crude protein) and isolipidic (19% crude lipids) diets were formulated to contain 20% waxy maize starch or 20% glucose. Triplicate groups of fish (63.5 g initial body weight) were fed each diet to near satiation during four weeks at 18 degrees C or 25 degrees C. Growth, feed intake, feed efficiency and protein efficiency ratio, were higher at the higher water temperature. At each water temperatures fish growth and feed efficiency were higher with the glucose diet. Plasma glucose levels were not influenced by water temperature but were higher in fish fed the glucose diet. Hepatosomatic index and liver glycogen were higher at the lower water temperature and within each water temperature in fish fed the glucose diet. No effect of water temperature on enzymes activities was observed, except for hexokinase and GK which were higher at 25 degrees C. Hepatic hexokinase and pyruvate kinase activities were not influenced by diet composition, whereas glucose-6-phosphate dehydrogenase activity was higher in fish fed the glucose diet. Higher GK activity was observed in fish fed the glucose diet. GK gene expression was higher at 25 degrees C in fish fed the waxy maize starch diet while in fish fed the glucose diet, no temperature effect on GK gene expression was observed. Hepatic G6Pase activities and gene expression were neither influenced by dietary carbohydrates nor water temperature. Overall, our data suggest that in gilthead sea bream juveniles hepatocytes dietary carbohydrate source and temperature affect more intensively GK, the enzyme responsible for the first step of glucose uptake, than G6Pase the enzyme involved in the last step of glucose hepatic release.     

Use of soybean flour (dehulled, solvent-extracted soybean) as a fish meal substitute in practical diets for African catfish, Clarias gariepinus (Burchell 1822): growth, feed utilization and digestibility

Growth and digestibility trials were conducted using African catfish, Clarias gariepinus (Burchell 1822): (1) to obtain apparent digestibility coefficient (ADC) values for capelin fish meal, soybean flour and corn meal; (2) to formulate diets based on ADC values of the protein feedstuffs; and (3) to evaluate the effects of replacing 25%, 50% and 75% of fish meal in control diets with soybean flour on growth, feed utilization efficiency and carcass composition. Supplemental methionine was added to the diet formulation in which soybean flour replaced 75% of the diet. Diets were formulated (400 g digestible protein kg⁻¹ and 15 kJ digestible energy g⁻¹ dry diet) and fed to catfish fingerlings (13.1 ± 0.5 g) to apparent satiation twice daily for 70 days. The protein and energy digestibilities of fish meal and soybean flour were high (>90% and >80%, respectively; P < 0.05). At 75% fish meal replacement with soybean flour (without methionine supplementation), growth and feed utilization efficiency indicators were depressed compared with other diet treatments which had a similar (P > 0.05) growth and feed utilization efficiency to those fed the control diet. The carcass compositions of catfish in all diets were similar (P > 0.05) and the liver histology of catfish fed any of the diets showed no alterations. The results obtained indicate that 50% of fish meal protein in practical catfish diets can be replaced with soybean flour and that catfish can effectively utilize supplemental methionine, thereby allowing up to 75% of the dietary fish meal protein to be replaced by soybean flour.     

Gene expression,enzyme activity and performance of Nile tilapia larvae fed with diets of different CP levels

Protein is the most costly nutrient in fish feed, and while diets offered in the early stages of development typically have high levels of CP, they do not always correspond to the real requirements of the animals. Thus, research that seeks to learn the true nutritional requirements of fish is fundamental to improving commercial fish culture. The present study evaluated the protein requirements of Nile tilapia (Oreochromis niloticus) under larviculture. Fish performance, gene expression for digestive enzymes and their enzymatic activity and stress response to air exposure were analyzed. Four experimental diets differing in CP level were formulated: 30%, 36%, 42% and 48%. Fish larvae were fed the experimental diets during development and sampled 10, 20 and 30 days after the beginning of the experiment for performance, gene expression and enzymatic activity. At sampling time 30, stress resistance was also evaluated by means of an air exposure test. At sampling time 10, CP levels between 36% and 48% could be used for a better performance. During this period, pepsinogen expression was greater for 30% CP, intermediate for 42% and lower for 36% and 48%. After this initial period, diets of between 30% and 42% CP are recommended for better performance. At sampling time 20, gene expression for digestive enzymes and their enzymatic activity were similar for all diets tested. At sampling time 30, the diet of 42% CP induced both greater pepsinogen expression and pepsin activity. Survival after the air exposure test after 30 days of feeding was influenced by CP level in the diet, with the highest survival being for fish fed with 36% CP. Taken together, the present results demonstrate that dietary CP influences digestive enzyme gene expression and activity, and suggest that the best CP levels for Nile tilapia larviculture vary depending on larval stage.     

氧化油脂对斑点叉尾鮰生长、体色和抗氧化能力的影响

为考察正常及氧化鱼油、豆油、混合油脂(鱼油﹕豆油=1﹕1)对斑点叉尾鮰(Ictalurus punctatus)生长性能、体色和肝脏抗氧化能力的影响, 设计6组等氮等脂饲料, 分别添加6%鱼油、6%豆油、3%鱼油+3%豆油、6%氧化鱼油、6%氧化豆油、3%氧化鱼油+3%氧化豆油, 饲喂6组初始体重(150.5±4.2 g)的斑点叉尾鮰8周, 每组3个重复, 每个重复14尾鱼。结果表明, 摄食6%鱼油、6%豆油、3%鱼油+3%豆油饲料的斑点叉尾鮰在增重率、饲料系数、肌肉组成、体色和肝脏抗氧化指标上均无显著差异(P>0.05); 摄食3组氧化油脂饲料后, 鱼体增重率和肌肉粗脂肪含量降低(P<0.05), 饲料系数和背部、尾部皮肤b*(黄度值)增加(P<0.05), 肝脏丙二醛(MDA)和血清谷丙转氨酶(ALT)、谷草转氨酶(AST)、总胆红素(TBIL)和直接胆红素升高(DBIL)(P<0.05), 肝脏超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、还原型谷胱甘肽降低(GSH)(P<0.05), 而肌肉色度值(L*、a* 、b*)和肌肉水分、粗蛋白质、灰分含量无显著差异(P>0.05)。以上结果表明, 6%氧化油脂(鱼油、豆油或混合油脂)导致斑点叉尾鮰生长性能下降、皮肤黄度增加、肝脏抗氧化能力受损; 豆油可替代斑点叉尾鮰饲料中鱼油的使用, 而不会对生长产生负面影响。     

饲料中糖/脂肪比例对瓦氏黄颡鱼生长、饲料利用、血糖水平和肝脏糖酵解酶活力的影响

以初始体重(4.20±0.02)g的瓦氏黄颡鱼(Pelteobagrus vachelli)幼鱼为实验对象,在流水系统中进行为期8周的摄食生长实验,探讨饲料中糖与脂肪比例(CHO∶L)对其生长、饲料利用、血糖水平和糖酵解酶活力的影响。饲料等氮(粗蛋白40%)等能(19 MJ/g),CHO∶L梯度为0.75—6.53。每种饲料随机投喂3桶鱼,每桶(50 L)放养40尾鱼。实验结果表明随着CHO∶L升高,瓦氏黄颡鱼特定生长率(SGR)先升高后降低,当CHO∶L为3.55时,SGR达最大,显著高于CHO∶L为0.75和6.53时的值(P<0.05)。饲料效率(FE)和蛋白效率(PER)在CHO∶L为1.30—3.55区间内都没有显著差异,但当CHO∶L为0.75或6.53时,FE和PER的值都显著降低(P<0.05)。脏体比(VSI)、肝体比(HSI)在各处理组间无显差异(P>0.05)。随CHO∶L的增加,全鱼粗蛋白含量先增加后降低,且在CHO:L为3.55时达到最大值,且显著高于CHO:L为0.75时的值(P<0.05)。而全鱼粗脂肪含量随CHO∶L增加而显著减少(P<0.05)。血糖和血浆总胆固醇水平在各处理组间无显著差异(P>0.05),而血浆甘油三酯水平随CHO∶L的增加而显著增加(P<0.05)。随CHO∶L的增加,瓦氏黄颡鱼肝脏葡萄糖激酶(GK)活力先增加后降低,且在CHO∶L为3.55时活力最大,显著高于CHO∶L为0.75时的值(P<0.05)。肝脏己糖激酶(HK)、丙酮酸激酶(PK)和磷酸果糖激酶(PFK-1)活力在各处理组间没有显著差异(P>0.05)。用二次多项回归模型拟合特定生长率和CHO∶L的关系,得到饲料最适CHO∶L为4.06。     

Effects of dietary soybean protein levels on energy budget of the southern catfish, Silurus meridionalis

Energy budgets were calculated for the southern catfish, Silurus meridionalis, fed diets replacing 0%, 13%, 26%, 39%, 52% and 65% fish meal protein with soybean meal (SBM) protein with or without methionine supplementation to apparent satiation at 27.5 degrees C. With increasing dietary soybean protein levels (SPL), the feed energy lost in feces, excretion and metabolism increased, while that available for growth decreased (P<0.05). When 0.12% or 0.26% methionine at 39% SPL was added to reach that in body carcass or the control group (0% SPL), no significant differences were found in each component of energy budgets. When 0.21% or 0.33% methionine at 52% SPL was added to reach the content of methionine in body carcass or the control group, energy spent on growth increased, but that on excretion and metabolism decreased (P<0.05). These results suggested that the differences in growth rate among the southern catfish fed the diets with different SPL were due to decreasing absorption rate, increasing excretion and metabolism with increasing dietary SPL. The most important factor limiting the use of soybean protein was the imbalance of essential amino acids, which resulted in more energy spent on metabolism and excretion, less energy on growth. Supplementation of methionine produced a relatively better amino acid profile and subsequently improved the utilization of soybean protein at high SPL, which resulted in less energy used for metabolism or lost in excretion and more energy available for growth.