Growth depression in socially subordinate rainbow trout Oncorhynchus mykiss: more than a fasting effect

To assess the effects of subordinate social status on digestive function, metabolism, and enzyme activity in salmonid fish, juvenile rainbow trout Oncorhynchus mykiss were paired with size-matched conspecifics (<1.5% difference in fork length) for 5 d. Fish that were fasted for 5 d and fish sampled directly from the holding tank were used as control groups. Both subordinate and fasted fish experienced significant decreases in intestine mass (P = 0.043), and the gall bladder showed marked and significant changes in both size (P = 0.004) and appearance. These findings suggest that the negative effect of social subordination on digestive function reflects in large part a lack of feeding. Hepatic phosphoenolpyruvate carboxykinase activity was significantly higher in subordinate fish relative to dominants, whereas subordinate hepatic pyruvate kinase activity was significantly lower; activities of both enzymes were significantly correlated with plasma cortisol concentrations and behavior scores. Dominant-subordinate differences in the activities of these enzymes were eliminated by administration of the glucocorticoid receptor antagonist RU486, underlining a role for circulating cortisol in eliciting the differences. Significant increases relative to control fish were also detected in red and white muscles from subordinate fish in the activities of protein catabolic enzymes (aspartate aminotransferase, alanine aminotransferase, glutamate dehydrogenase). These differences occurred in the absence of any change in plasma free amino acid or ammonia concentrations, supporting an enhanced turnover of amino acids in muscle in subordinate fish. The results support the hypothesis that changes in metabolism, beyond those elicited by low food consumption, may be responsible at least in part for the low growth rates typical of subordinate fish and that these changes may be related specifically to circulating cortisol levels in subordinate fish.     

晶体氨基酸替代鱼粉蛋白对半滑舌鳎稚鱼消化酶和代谢酶活力的影响

为研究人工微颗粒饲料中晶体氨基酸替代鱼粉蛋白对半滑舌鳎（Cynoglossus semilaevis Gnther）稚鱼消化酶和代谢酶活力的影响, 以晶体氨基酸混合物分别替代0%、25%、50%、75%和100%鱼粉蛋白（0%CAA、25%CAA、50%CAA、75%CAA和100%CAA）, 在25%替代水平设计棕榈酸甘油酯包被晶体氨基酸混合物组（C-25%CAA）, 配制实验微颗粒饲料。每种微颗粒饲料随机投喂三组实验鱼初始体重（0.0940.02） g, 35日龄, 每组实验鱼放养150尾, 养殖周期28d。研究结果表明, 在不包膜条件下, 各处理组的胰蛋白酶活力随替代水平的升高显著下降（P0.05）, 且包膜处理组（C-25%CAA）与全鱼粉组无显著差异（P0.05）。肠段胰蛋白酶活力与胰段胰蛋白酶活力比值随氨基酸替代水平的升高显著下降（P0.05）, 鱼粉组显著高于75%和100%处理组（P0.05）, 但与25%处理组、包膜处理组（C-25%CAA）和50%处理组无显著差异（P0.05）。各处理组淀粉酶活力随替代水平的升高显著上升（P0.05）。亮氨酸氨肽酶（LA）和碱性磷酸酶（AP）活力（肠段与刷状缘）均随替代水平的升高显著下降（P0.05）, 包膜处理组（C-25%CAA）与全鱼粉组无显著差异（P0.05）。谷丙转氨酶（GPT/ALT）和谷草转氨酶（GOT/AST）活力随替代水平的升高显著上升（P0.05）, 50%、75%和100%处理组显著高于鱼粉组、25%处理组和包膜处理组（C-25%CAA）。研究结果显示, 饲料中晶体氨基酸显著影响了半滑舌鳎稚鱼的消化酶和代谢酶活力, 而且在25%的替代水平下, 与未包膜组相比, 包膜处理组能显著促进半滑舌鳎稚鱼消化系统的发育。       

Effect of dietary fish on antioxidant parameters of normal and cholesterol stressed rats

Feeding fish (Sardinella longiceps) to normal rats increased lipid peroxidation and total and Se-dependent glutathione peroxidase (GSH-px) activity in erythrocytes and manganese dependent superoxide dismutase (Mn-SOD) activity in liver. Feeding fish to cholesterol stressed rats showed a significant increase in the activity of GSH-px and cholesterol feeding alone, resulted in a significant increase in the lipid peroxidation and liver Mn-SOD activity. The results suggest that the high polyunsaturated fatty acid content of S. longiceps, the fish abundantly available in the west coast of India, does not have any deleterious effect by way of free radical generation. The observed lipid peroxidation is not critical as is evident from the results of glutathione level and other scavenging enzymes.     

Cd2+急性胁迫下方格星虫体腔液消化酶活力和游离氨基酸的变化

为了解Cd2+急性胁迫下方格星虫体腔液消化酶活力和游离氨基酸的变化规律, 采用毒理学试验方法, 在确定Cd2+对方格星虫毒性强度的基础上, 选取48h最低致死浓度为试验浓度, 研究该浓度Cd2+胁迫下方格星虫体腔液消化酶活力和游离氨基酸的动态变化。结果表明: 方格星虫死亡率随着Cd2+浓度的升高而增加,Cd2+对方格星虫的24h和48h的LC50分别为37.80和22.68 mg/L。在48h最小致死浓度下, Cd2+对方格星虫体腔液蛋白酶和淀粉酶活力在试验周期内均表现为抑制, 且淀粉酶活力受到的抑制作用较强; Cd2+胁迫前期脂肪酶活力显著升高(P0.05), 24h后又显著降低(P0.05), 48h时仅为初始水平的40%, 说明低浓度Cd2+对脂肪酶活力有诱导作用, 高浓度Cd2+则产生抑制。方格星虫体腔液游离氨基酸的组成和含量在Cd2+胁迫48h内均有显著变化(P0.05)。各游离氨基酸含量及氨基酸总量在24h前均无显著变化(P0.05), 24h后先上升后下降(P0.05),36h的游离氨基酸总量达到初始水平的2倍以上, 为145.50 mg/100 mL, 大部分游离氨基酸组成百分比也在24h前较稳定, 24h后呈现峰值变化。总之, Cd2+急性胁迫对方格星虫体腔液消化酶活力和游离氨基酸均有显著影响(P0.05), 且消化酶活力与游离氨基酸含量和组成的变化与胁迫时间有关。     

烧伤患者血浆和红细胞游离氨基酸变化及输注氨基酸对其变化的影响

动态测定烧伤患者血浆及红细胞内游离氨基酸的含量 ,探讨输入外源性氨基酸后对血及红细胞内游离氨基酸的影响。以日立 835— 5 0型氨基酸自动分析仪测定烧伤患者血浆及红细胞内游离氨基酸含量。结果发现烧伤患者血浆总游离氨基酸浓度从伤后到 2 1天均显著降低 (P <0 .0 5～ 0 .0 1) ;赖、苯丙和苯丙 酪氨酸比值显著升高 (P <0 .0 5～ 0 .0 1) ;色、组、精、丙、甘、苏、脯和丝氨酸比值显著降低 (P <0 .0 5～ 0 .0 1) ;缬、亮、异亮、酪、胱和支链氨基酸伤后早期降低。烧伤患者红细胞内总游离氨基酸含量不同程度降低 ,其中 1、3、7天降低显著 (P <0 .0 5～ 0 .0 1) ;红细胞内苯丙和苯丙 酪氨酸比值未见显著性升高 ;色、蛋、精、脯氨酸含量很低或基本未测出。输注复合氨基酸注射液后未能显著改善患者血及红细胞内游离氨基酸含量。结果提示烧伤患者红细胞内游离氨基酸含量的变化趋势与血浆游离氨基酸变化趋势基本一致 ;烧伤后红细胞内苯丙氨酸及苯丙 酪氨酸比值有别于血浆变化。本研究条件下补充外源性氨基酸未能显著改变烧伤患者血浆及红细胞内游离氨基酸的含量     

Assemblies of free amino acids as possible prebiotic catalysts

Our understanding of how life emerged on Earth has much to do with speculations about the ways in which prebiotic catalysts could have been formed. Since enzymes, the contemporary biological catalysts, are polymers of amino acids, we looked at the possible activity of free amino acids as catalysts. In this study it is shown experimentally that mixtures of free amino acids exert catalytic activities of -galactosidase, carbonic anhydrase, and catalase. We also observed different levels of catalytic activty of individual amino acids: some were more efficient than others. Apparently, assemblies of amino acids which were formed around substrate molecules through weak interactions, could, in principle, catalyze many prebiotic reactions. This might have been one step in the emergence of biological enzymes.     

ALTERATION OF GABA SYSTEM AND PURKINJE CELLS IN RAT CEREBELLUM BY THE CONVULSANT 3-MERCAPTOPROPIONIC ACID

Abstract— The effect of the convulsant, 3-mercaptopropionic acid (MP) on the content of free amino acids and on the activity of some enzymes related to their metabolism was studied in the rat cerebellum. A decrease in the activity of glutamate decarboxylase (EC 4.1.1.15) and in the level of GABA was found; at the same time, the activity of GABA-aminotransferase (EC 2.6.1.19) was increased. These changes coincided with a profound alteration of the morphology of the Purkinje cells which was related to the dose of MP. These findings, plus some changes in the content of other free amino acids and the activities of related enzymes, suggest that 3-mercaptopropionic acid induces in the cerebellum an imbalance among the amino acids involved in the excitation-inhibition mechanisms.     

Pancreatic and intestinal carbohydrases are matched to dietary starch level in wild passerine birds

Evolutionary shifts in diet composition are presumably accompanied by simultaneous changes in digestive physiology. The adaptive modulation hypothesis predicts that activities of digestive enzymes should match the relative levels of their substrates in an animal's diet so that available membrane space and synthetic energy are not wasted on enzymes in excess of need. However, previous studies on captive passerine birds showed high intraspecific phenotypic flexibility only in proteases but not in carbohydrases in response to varying diet composition. In this study, we measured the activities of pancreatic, intestinal, and hepatic enzymes in six wild-caught passerine species. We predicted that if the adaptive modulation hypothesis holds during evolutionary shifts in diet composition in birds, then mass-specific activities of digestive enzymes should be correlated positively with the content of their relevant substrates in species' diets. Whereas mass-specific activities of proteases (aminopeptidase-N, trypsin, chymotrypsin, alanine aminotransferase) were not correlated with estimated dietary protein content, mass-specific activities of all studied carbohydrases (amylase, maltase, sucrase) were positively correlated with estimated dietary starch content. We conclude that activities of carbohydrases but not proteases are evolutionarily matched to diet composition in passerine birds. We hypothesize that the need for nitrogen and essential amino acids can prevent the evolution of a low activity of proteases, even in species feeding on a low-protein diet.     

Responses of the pancreatic digestive enzyme secretion to amino acids, glucose and cholecystokinin in chicks

1. Pancreatic secretion of digestive enzymes, amylase, trypsinogen and chymotrypsinogen, was studied in response to the wing vein injection of digestive end products, various single amino acids and glucose, with or without cholecystokinin (CCK) in chicks. 2. Among amino acids administered, only phenylalanine significantly (P less than 0.05) increased trypsinogen and chymotrypsinogen secretions, while other amino acids did not. 3. Simultaneous injection of single amino acids with CCK increased digestive enzyme secretion to various extents depending on the kind of amino acids whereas the injection of glucose with CCK did not affect when compared with that of CCK alone. 4. By varying doses, a synergetic action of CCK plus amino acid on the secretion of pancreatic digestive enzymes was observed at 0.5 mM for valine and 5 mM for arginine.     

烧伤对兔血红细胞内游离氨基酸浓度的影响

动态观察烧伤兔血红细胞内游离氨基酸的代谢变化。以日立 835 - 5 0型氨基酸自动分析仪测定 30 %TBSA兔烧伤后 1、3、7、15和 2 0d血红细胞内游离氨基酸的含量。结果显示 ,烧伤后多数测试点红细胞内游离氨基酸不同程度降低 ;伤后 1天降低最显著 ;苏、丙、缬、组、精和脯氨酸伤后各个时间点降低 ,多数时间点显著降低 (P <0 .0 5 - 0 .0 1)。结果提示烧伤后红细胞内游离氨基酸依靠易化扩散参与全身氨基酸的再分布     

Free amino acids and some enzymes of placental amino acid metabolism at different stages of its development]

There were revealed changes in the content of free amino acids and in the activity of glutamic dehydrogenase, aspartic- and alanine-aminotransferases in the course of development of the placenta in guinea pigs. The greatest values by the indices under study were reached by the 25th--30th days of pregnancy, i.e. by the period of final formation of the placenta. By the 40th day of pregnancy the activity of the enzymes and the content of free amino acids fell, and from the 45th day persisted at a certain stable level up to the occurrence of labour.     

Digestive enzymes and metabolic profile of Labeo rohita fingerlings fed diets with different crude protein levels

Labeo rohita, commonly called rohu is one of the most important fish species for aquaculture in India. Digestive enzyme response and metabolic profile of fingerling L. rohita to different dietary crude protein (CP) levels (viz. 25, 30, 35 and 40%) were studied in an attempt to optimize a practical diet formulation for this species. After 45 days of feeding, activity of digestive enzymes and metabolite concentrations were assayed. Amylase, lipase and alkaline phosphatase (ALP) activities were not influenced by the dietary protein, but proteolytic and acid phosphatase (ACP) activities varied (P<0.05) between the treatments. Proteolytic activity showed a second order polynomial relationship with dietary crude protein (CP) as Y = 0.0734X(2) + 4.937X - 68.37, r(2)=0.97. A positive correlation was observed between dietary CP and amylase (r(2)=0.78). All the metabolites except muscle glucose showed significant change corresponding to the dietary protein levels. Glucose and glycogen levels corresponded to the dietary carbohydrate levels. Muscle and plasma pyruvic acid increased as the crude protein in the diet increased, whereas liver pyruvic acid showed the opposite trend. Muscle protein content was not affected by dietary CP. Protein fractions in plasma (total protein, albumin and globulin) showed maximum values in 30% CP fed group. It is concluded that proteolytic activity and ACP are the major digestive enzymes responsive to dietary CP in L. rohita fingerlings. Considering the cost effectiveness of the diet, and based on liver and plasma free amino acid levels and plasma protein fractions, 30% crude protein is recommended as the optimal dietary protein for L. rohita fingerlings.     

Changes in metabolism and hepatic ultrastructure induced by estradiol and testosterone in immature female Epinephelus akaara (Teleostei,Serranidae)

Summary Estradiol injections increase serum level of calcium, amino acid, glucose, protein, ammonia and creatinine in immature Epinephelus akaara, and also increase levels of total lipid, cholesterol, phospholipid and esterified fatty acids. Hepatic protein, glycogen and lipid concentrations also rise after estradiol treatment, and some hepatic enzymes participating in the metabolism of nitrogen, lipid and carbohydrate, show increased activity. Serum vitellogenin levels are increased. Testosterone treatment increases serum protein, total lipid, cholesterol, amino acid and ammonia levels, and also hepatic glycogen content, but in contrast to estradiol treatment, testosterone does not change serum vitellogenin, glucose, calcium, phospholipid, esterified fatty acid and creatinine levels, nor the hepatic lipid and protein content. A small number of hepatic enzymes shows an increased activity. Vitellogenic fish show biochemical changes similar to that of estradiol-treated fish, but are different from those of immature fish. Estradiol treatment induces ultrastructural changes in the hepatocytes of immature fish that are similar to those found in vitellogenic fish. These include a proliferation of rough endoplasmic reticulum and Golgi apparatus, and an increase in glycogen and lipid, all indicative of enhanced metabolic activity.     

Protein digestion and ion concentrations in rainbow trout (Salmo gairdnerii Rich.) digestive tract in sea- and fresh water

• 1.1. Rainbow trout maintained in fresh water or Actapted to sea-water for 24 hr were fed casein-based dry diet. After feeding, fish were kept in fresh water (FW) or transferred to artificial sea-water (SW) and sacrificed after 10 or 20 hr.
• 2.2. The digestive tract was separated into five parts: stomach, pyloric caeca region, middle intestine and two equal lengths of rectum.
• 3.3. The content of these parts was analysed for ions Na⁺, K⁺, Cl⁻, Mg²⁺ and for free, peptide and total amino acids.
• 4.4. In the fish stomach all ions, with the exception of Ca²⁺, indicate drinking of sea-water. In the pyloric caeca region Na⁺ appears to be efficiently absorbed in SW fish but influxed in FW fish. In the rectum of SW fish K⁺ appears to be reabsorbed but Na⁺ concentrated in faeces.
• 5.5. Free amino acid concentrations were always higher in gut lumen of SW than in FW fish in respect to time after feeding and portion of intestinal content. Free amino acids constitute at most 7.4–8.7% of total amino acids in the content of pyloric caeca region.
• 6.6. Peptide amino acids, being mostly di-, tri- and tetra-peptides, increased in stomach content from 14.7 to 28.4% of the total, from 6 to 10 hr after a meal in SW fish. Peptide amino acids constituted 80.3–89.0% of total amino acids in intestinal content of the pyloric caeca region. These peptide portions decreased in the mid-intestine (47.5–52.5%) and increased again in the rectum (73.6–76.0%).
• 7.7. It was concluded that in rainbow trout fed in both sea- or fresh water, ion concentrations do not seem to interfere with protein digestion and nutrient absorption in alimentary tract.

     

Influence of dietary non-essential amino acid profile on growth performance and amino acid metabolism of Nile tilapia, Oreochromis niloticus (L.)

The quality of dietary protein is an important factor influencing the growth performance of fish. To evaluate the quality of protein, the variables commonly studied are the composition of the essential amino acids, the digestibility and the protein use efficiency. The goal of the present experiment was to test the effect of the dietary non-essential amino acid composition on the growth of Nile tilapia (Oreochromis niloticus). The fish were fed three purified diets differing only in their non-essential amino acid composition. The influence of the experimental diets on the growth performance, on the activity of enzymes involved in the amino acid metabolism, aspartate aminotransferase (ASAT) and alanine aminotransferase (ALAT), and on whole body delta(15)N values was investigated. Body mass, lipid, protein and energy gain differed significantly between the feeding groups. The activity of ASAT in the whole liver was significantly higher in fish with a positive protein balance compared to fish which lost protein. Whole body delta(15)N values of fish were negatively correlated with their body mass gain. Despite the poor utilisation of synthetic amino acids, the experiment indicates the importance of the dietary non-essential amino acid composition for the growth performance of fish. The study reveals the possibility to trace the utilisation of synthetic amino acids by determining the isotopic composition of dietary amino acids and tissues or whole bodies of animals.     

Studies on the role of exogenous proteolytic enzymes in digestion processes in fish

Studies were carried out on the proteolytic activity of the fry of common carp, rainbow trout, grass carp, and whitefish, as well as on the activity of digestive organs of adult common carp and rainbow trout. Activity of exogenous enzymes in relation to endogenous ones was assessed on the basis of the proteolytic value of fish food and the activity of digestive organs. It was found that the share of proteolytic enzymes of natural food in the digestion process in fish was high. Beginning from a weight of 50–100 g for common carp and 10 g for rainbow trout, the relation between the daily enzymatic ration and the weight of fish indicates the cooperation of an approximately constant amount of exogenous enzymes.     

Digestive enzymes during ontogeny of the sturgeon Acipenser naccarii: intestine and pancreas development

The present work examines the temporal appearance and degree of activity of digestive enzymes of pancreatic and intestinal origin (amylase as well as neutral and alkaline proteases) together with the evolution of the ontogenic development of the intestine and pancreas during the first month of free‐living (30 days post‐hatch–[dph]) of the sturgeon Acipenser naccarii. In addition, the influence of exogenous live feeding with Artemia salina on the detection of digestive enzymes was studied. Shown are that in the first life stages of the A. naccarii, from the time of fertilisation to the juvenile stage, the following events related to its digestive physiology occurred: digestive enzymes were detected in the embryo stage within the egg, presumably related to the hatching gland; in the free‐living embryo stage, opening of the mouth was at 10 dpf and protease and amylase digestive activities occurred due to an early differentiation of the pancreas and intestine (12–13 dpf). At this stage, digestive enzymatic activities are partly owing to the live feed offered before disappearance of the yolk reserves (14 dpf); the expulsion of the melanin plug and the appearance of a continuity of the digestion tract (16 dpf) together with the disappearance of the yolk reserves (17 dpf) mark the passage from the free‐living embryo phase to the juvenile stage; finally, from the first month after hatching in this sturgeon species, there is a stabilisation in digestive amylase and protease activities together with a fully developed digestive structure. The contribution of enzymes from prey in the detection of enzymatic activities determined in the fish was beyond doubt; therefore these exogenous enzymes must also have a certain physiological importance. Subsequent work will be needed to clarify the importance of digestive processes of the exogenous enzyme pool from live feed as well as to assess the possibility of shortening the weaning period.     

Spontaneous degradation of hemoglobin and egg albumin

Hemolysates of red cells of the horse, after removal of stroma and prolonged dialysis, released free amino acids on standing. A similar release of amino acids was not found when a solution of purified hemoglobin A was allowed to age, but a small number of peptides were found. Crude and purified egg albumin, when allowed to stand in solution, released free amino acids in a manner similar to that of the hemolysate of red cells. Quantitative data on the release of free amino acids are reported in this communication. The variety and distribution of the amino acids make it unlikely that this hydrolytic process is solely the result of contamination by proteolytic enzymes. The proposal is made that this property results from an intrinsic enzyme-like activity associated with the sulfhydryl groups of these proteins.