Effect of dietary n-3 fatty acids on HMG-CoA reductase and ACAT activities in liver and intestine of the rabbit

The regulation of hepatic and intestinal 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) reductase and acyl-CoA; cholesterol acyltransferase (ACAT) activities by dietary fish oil was examined in the rabbit. Rabbits were fed 10% menhaden oil or menhaden oil plus 1% cholesterol for 14 days. They were compared with animals fed a control diet or one enriched with long-chain saturated fats consisting of 10% cocoa butter oil or cocoa butter oil plus 1% cholesterol. Plasma cholesterol was increased in rabbits fed the fish oil and the two cholesterol-containing diets. In the liver, ACAT activity was increased and HMG-CoA reductase activity was decreased in rabbits ingesting the fish oil. The same was true for animals ingesting both cholesterol-containing diets. In the intestine, ACAT activity was not affected by the ingestion of the fish oil compared to control rabbits; however, it was significantly higher in animals fed the fish oil compared to animals ingesting the cocoa butter. HMG-CoA reductase activity was decreased in the distal two-thirds of the intestine in animals fed the menhaden oil compared to activities observed in controls. In animals ingesting the cholesterol diets, intestinal reductase was significantly decreased, whereas intestinal ACAT activity was increased in rabbits ingesting the cocoa butter and cholesterol diet when compared to their controls. Lipid analysis of hepatic and intestinal microsomes demonstrated an enrichment of n-3 polyunsaturated fatty acids in membranes from rabbits ingesting the menhaden oil.(ABSTRACT TRUNCATED AT 250 WORDS)     

Effects of diets containing soybean meal on trypsin mRNA expression and activity in Atlantic salmon (Salmo salar L)

Atlantic salmon develop subacute enteritis in the distal intestine (DI) when fed diets containing soybean meal (SBM) at high levels, a condition accompanied by increased trypsin activity in the DI intestinal content compared to fish fed conventional fishmeal (FM) based diets. To further investigate the responses of Atlantic salmon to dietary SBM, we measured trypsin activity in intestinal contents, quantified pancreatic trypsin mRNA expression, surveyed trypsin mRNA expression in selected tissues and characterized active forms of trypsin in the intestinal wall and brain. Enzyme measurements showed that trypsin activity in the intestinal content of SBM fed fish was lower in the proximal segments of the intestine, but higher in the DI compared to FM fed fish. The difference in enzyme activity was not reflected in a differential expression of pancreatic trypsin mRNA between fish fed the different diets (FM or SBM). Trypsin mRNA was expressed in 18 different tissues (esophagus, stomach, pancreas, pyloric tissue, midintestine, distal intestine, liver, head kidney, kidney, heart, spleen, thymus, brain, eye, gills, gonads, muscle and skin) but was most prominently expressed in tissues of the gastrointestinal (GI) tract and brain. We report for the first time an upregulation of trypsin-like activity in the DI wall using an in-gel trypsin activity assay, as well as modulated activity in the brain of fish fed SBM. The increased activity in the DI wall may contribute to disease severity and higher trypsin activity in the intestinal content.     

Aquacultured Rainbow Trout (Oncorhynchus mykiss) Possess a Large Core Intestinal Microbiota That Is Resistant to Variation in Diet and Rearing Density

As global aquaculture fish production continues to expand, an improved understanding of how environmental factors interact in fish health and production is needed. Significant advances have been made toward economical alternatives to costly fishmeal-based diets, such as grain-based formulations, and toward defining the effect of rearing density on fish health and production. Little research, however, has examined the effects of fishmeal- and grain-based diets in combination with alterations in rearing density. Moreover, it is unknown whether interactions between rearing density and diet impact the composition of the fish intestinal microbiota, which might in turn impact fish health and production. We fed aquacultured adult rainbow trout (Oncorhynchus mykiss) fishmeal- or grain-based diets, reared them under high- or low-density conditions for 10 months in a single aquaculture facility, and evaluated individual fish growth, production, fin indices, and intestinal microbiota composition using 16S rRNA gene sequencing. We found that the intestinal microbiotas were dominated by a shared core microbiota consisting of 52 bacterial lineages observed across all individuals, diets, and rearing densities. Variations in diet and rearing density resulted in only minor changes in intestinal microbiota composition despite significant effects of these variables on fish growth, performance, fillet quality, and welfare. Significant interactions between diet and rearing density were observed only in evaluations of fin indices and the relative abundance of the bacterial genus Staphylococcus. These results demonstrate that aquacultured rainbow trout can achieve remarkable consistency in intestinal microbiota composition and suggest the possibility of developing novel aquaculture strategies without overtly altering intestinal microbiota composition.     

Distribution of metals during digestion by cutthroat trout fed benthic invertebrates contaminated in the Clark Fork River, Montana and the Coeur d'Alene River, Idaho, U.S.A., and fed artificially contaminated Artemia

The concentrations of essential amino acids in three, undigested invertebrate diets collected from the Clark Fork River (CFR) for cutthroat trout were similar to each other, but were c. 25–75% less than Artemia that were exposed to a mixture of arsenic, copper, cadmium, lead and zinc in the laboratory. The Artemia diet appeared less palatable and the texture, quantity and appearance of the intestinal contents differed between fish fed the Artemia and CFR diets. The Pb% in the fluid fraction of the intestinal contents was greater for the Artemia (29%) than for the CFR diets (10–17%), and the Cu% in the amino acid plus metal fraction of the intestinal contents was greater for the Artemia (78%) than for two of the three CFR diets (67% and 70%). Intestinal contents of fish fed invertebrate diets collected from various sites on the Coeur d'Alene River (CDA), Idaho, were similar in texture, quantity, and appearance. For fish fed the CDA diets, differences in the distribution of metals among fractions of the digestive fluids appeared to be related to concentrations of metals in the invertebrate diets. Pb% was lowest of all metals in the fluid portion of the intestinal contents. However, >80% of all metals in the hind gut were associated with the particulate fraction where they may still be available for uptake through pinocytosis.     

Histological alterations and microbial ecology of the intestine in gilthead seabream (Sparus aurata L.) fed dietary probiotics and microalgae

The effects on histology and microbial ecology in gilthead seabream (Sparus aurata) intestine caused by dietary probiotic and microalgae were studied. Fish were fed non-supplemented (C, control) or supplemented diets with Tetraselmis chuii, Phaeodactylum tricornutum and Bacillus subtilis single or combined (diets T, P, B, BT and BP) for 4?weeks. Curiously, fish fed the experimental diets showed similar morphological alterations when studied by light and electron microscopy and significant signs of intestinal damage were detected. No effect of microalgae or B. subtilis on the intestinal absorptive area was observed, whereas the number of goblet cells and IELs were significantly lower in fish fed the T, P, B and BT diets and T, BT and BP diets, respectively. Interestingly, only the diets containing B. subtilis resulted in a significant reduction of microvilli height. Alterations such as wide intercellular spaces and large vacuoles in enterocytes were observed in fish fed T, B, BT, BT and P in lesser degrees. These observations demonstrate that fish fed experimental diets presented different signs of oedema and inflammation that could compromise their body homeostasis. Moreover, the experimental diets cause important alterations in the intestinal microbiota by a significant decrease in bacterial diversity, as demonstrated by the fall in specific richness, Shannon and range-weighted richness indices. To our knowledge, this is the first in vivo study regarding the implications of the use of probiotics in combination with immunostimulants on fish intestinal morphology and microbiota. More morphofunctional studies are needed in order to correlate the nutritional and immune aspects of fish gut.     

Use of alternative protein sources for fishmeal replacement in the diet of largemouth bass (Micropterus salmoides). Part I: effects of poultry by-product meal and soybean meal on growth,feed utilization,and health

Five isonitrogenous and isocaloric diets [containing 54, 30, 15, 10, and 5% fishmeal crude-protein (CP), dry matter (DM) basis] were prepared by replacing fishmeal with poultry by-product meal plus soybean meal to feed juvenile largemouth bass (LMB, with an initial mean body weight of 4.9 g) for 8 weeks. All diets contained 54% CP and 13% lipids. There were four tanks of fish per treatment group (15 fish/tank). The fish were fed twice daily with the same feed intake (g/fish) in all the dietary groups. Results indicated that the inclusion of 15% fishmeal protein in the diet is sufficient for LMB growth. However, some of the fish that were fed diets containing ≤ 15% fishmeal CP had black skin syndrome (characterized by skin darkening and retinal degeneration, as well as intestinal and liver atrophies and structural abnormalities). The concentrations of taurine, methionine, threonine and histidine in serum were reduced (P < 0.05) in fish fed the diets containing 5, 10 and 15% fishmeal CP, compared with the 30 and 54% fishmeal CP diets. Interestingly, the concentrations of tyrosine and tryptophan in serum were higher in fish fed diets with ≤ 15% fishmeal CP than those in the 54% fishmeal CP group. These results indicated that 15% fishmeal CP in the diet containing poultry by-product meal and soybean meal was sufficient for the maximum growth and feed efficiency in LMB but inadequate for their intestinal, skin, eye, and liver health. A reduction in dietary methionine and taurine content and the possible presence of antinutritional factors in the fishmeal replacements diets containing high inclusion levels of soybean meal may contribute to black skin syndrome in LMB. We recommend that the diets of juvenile LMB contain 30% fishmeal CP (DM basis).

     

Enzyme activities of intestinal triacylglycerol and phosphatidylcholine biosynthesis in Atlantic salmon (Salmo salar L.)

The substitution of fish oil with plant-derived oil in diets for carnivorous fish, such as Atlantic salmon, has previously revealed the potentially deleterious supranuclear accumulation of lipid droplets in intestinal cells (enterocytes) which may compromise gut integrity, and consequently, fish health. This suggests that unfamiliar dietary lipid sources may have a significant impact on intestinal lipid metabolism, however, the mode of lipid resynthesis is largely unknown in teleost fish intestine. The present study aimed at characterising three key lipogenic enzymes involved in the biosynthesis of triacylglycerol (TAG) and phosphatidylcholine (PC) in Atlantic salmon enterocytes: monoacylglycerol acyltransferase (MGAT), diacylglycerol acyltransferase (DGAT), and diacylglycerol cholinephosphotransferase (CPT). Furthermore, to investigate the dietary effect of plant oils on these enzymes, two experimental groups of fish were fed a diet with either capelin (fish oil) or vegetable oil (rapeseed oil:palm oil:linseed oil, 55:30:15 w/w) as the lipid source. The monoacylglycerol (MAG) pathway was highly active in the intestinal mucosa of Atlantic salmon as demonstrated by MGAT activity (7 nmol [1-(14)C]palmitoyl-CoA incorporated min(-1) mg protein(-1)) and DGAT activity (4 nmol [1-(14)C]palmitoyl-CoA incorporated min(-1) mg protein(-1)), with MGAT appearing to also provide adequate production of sn-1,2-diacylglycerol for potential utilisation in PC synthesis via CPT activity (0.4 nmol CDP-[(14)C]choline incorporated min(-1) mg protein(-1)). Both DGAT and CPT specific activity values were comparable to reported mammalian equivalents, although MGAT activity was lower. Nevertheless, MGAT appeared not to be the rate-limiting step in salmon intestinal TAG synthesis. The homology between piscine and mammalian enzymes was established by similar stimulation and inhibition profiles by a variety of tested cofactors and isomeric substrates. The low dietary n-3/n-6 PUFA ratio presented in the vegetable oil diet did not significantly affect the activities of MGAT, DGAT, or CPT under optimised assay conditions, or in vivo intestinal mucosa lipid class composition, when compared to a standard fish oil diet.     

果胶高载引起的黄颡鱼绿肝症和肝纤维化与肠道菌群失调的关联

日粮中黏性膳食纤维果胶较高时可引起黄颡鱼(Pelteobagrus fulvidraco)的绿肝症和肝纤维化, 研究旨在探讨肠道菌群在上述病理过程中的潜在作用。分别配制含30%糊精和果胶的对照日粮和高果胶日粮, 命名为CON和PEC日粮, 再在CON和PEC日粮中添加0.2%的抑菌剂甲硝唑, 命名为CONM和PECM日粮。用4种日粮饲养初始体质量为(21.5±3.6) g/尾的黄颡鱼, 分别于饲养7d和56d后采样分析。16S rRNA分析结果显示, 日粮中添加甲硝唑抑制了梭杆菌门(Fusobacteria)和厚壁菌门(Firmicutes)的细菌, 使变形菌门(Proteobacteria)成为第一优势菌, 表明甲硝唑发挥了抑菌作用。饲养7d后CON组、CONM组、PEC组和PECM组的绿肝率分别为0、12%、27%和88%, 饲养56d后各组鱼肝脏纤维化程度依次增强。甲硝唑显著降低了肠道内容物中胆盐水解酶活力。肝脏中参与胆汁酸合成、转运及调控的基因表达活性与绿肝率及血清中胆汁酸含量不一致。上述结果提示肠道菌群紊乱可能是引起绿肝症和肝纤维化的重要原因, 胆汁酸可能介导了此病理过程。     

Effects of water temperature and diets containing palm oil on fatty acid desaturation and oxidation in hepatocytes and intestinal enterocytes of rainbow trout (Oncorhynchus mykiss)

Food grade fisheries have reached their sustainable limits while aquaculture production has increased to meet consumer demands. However, for growth in aquaculture to continue and utilise sustainable, feeding ingredients, alternatives to fish oil (FO), the predominant lipid component of fish diets, must be developed. Therefore, there is currently considerable interest in the regulation of fatty acid metabolism in fish in order to determine strategies for the best use of plant oils in diets for commercially important cultured fish species. Plant oils are characteristically rich in C18 polyunsaturated fatty acids (PUFA) but devoid of C20 and C22 highly unsaturated fatty acids (HUFA) found in FO. The fatty acyl desaturase enzyme activities involved in the biosynthesis of HUFA from PUFA are known to be under nutritional regulation and can be increased in fish fed diets rich in plant oils. However, fatty acid desaturase activity is also known to be modulated by water temperature in fish. The present study aimed to investigate the interaction between water temperature and diet in the regulation of fatty acid metabolism in rainbow trout. Trout, acclimatized to 7, 11 or 15 degrees C, were fed for 4 weeks on diets in which the FO was replaced in a graded manner by palm oil. At the end of the trial, fatty acyl desaturation/elongation and beta-oxidation activities were determined in isolated hepatocytes and intestinal enterocytes using [1-14C]18:3n-3 as substrate, and samples of liver were collected for analysis of lipid and fatty acid composition. The most obvious effect of temperature was that fatty acid desaturation/elongation and beta-oxidation were reduced in both hepatocytes and intestinal enterocytes from fish maintained at the highest water temperature (15 degrees C). There were differences between the two tissues with the highest desaturation/elongation and beta-oxidation activities tending to be in fish held at 11 degrees C in the case of hepatocytes, but 7 degrees C in enterocytes. Correlations between fatty acid metabolism and dietary palm oil were most clearly observed in desaturation/elongation activities in both hepatocytes and enterocytes at 11 degrees C. The highest beta-oxidation activities were generally observed in fish fed FO alone in both hepatocytes and enterocytes with palm oil having differential effects in the two cell types.     

低鱼粉饲料中添加苏氨酸对三倍体虹鳟抗氧化能力、消化生理及肠道炎症因子基因表达的影响

研究低鱼粉饲料中不同水平苏氨酸对三倍体虹鳟(Oncorhynchus mykiss)生长、抗氧化能力、消化生理及肠道炎症因子基因表达的影响, 以初始体质量(18.42±0.20) g的三倍体虹鳟为研究对象, 在室内200 L流水水族箱中进行摄食生长试验56d。在基础饲料中添加L-苏氨酸, 配制苏氨酸水平分别为0.45%(对照)、0.76%、1.09%、1.29%和1.64%的5种等氮等能饲料。将450尾三倍体虹鳟随机分为5组, 每组处理3个重复, 每个重复30尾鱼。结果表明, 当添加量为1.09%、1.29%和1.64%时, 血清超氧化物歧化酶(SOD)活力显著提高, 同时显著降低了丙二醛(MDA)的含量(P<0.05), 过氧化氢酶(CAT)在1.29%组含量最高, 相比对照组差异显著(P<0.05)。在饲料中添加苏氨酸(0.76%—1.64%)时, 肠道脂肪酶(LPS)和胰蛋白酶的消化酶活性显著高于对照组(P<0.05), 但随着饲料苏氨酸水平达到1.64%时, 消化酶活性有所降低。1.09%和1.65%组的肠道组织形态最佳, 绒毛发达, 排列整齐, 无融合和脱落现象。苏氨酸水平对肠道炎症因子IL-2、IL-8、IL-10、IgM、TNF-α和PepT1都有显著影响(P<0.05), 随着苏氨酸水平的增加, IL-2、IgM和PepT1呈现先升高后降低的变化趋势, 在1.29%和1.09%组的表达量达到最高; IL-8呈逐渐升高趋势; 而IL-10和TNF-α呈下降的变化趋势。研究结果表明, 在低鱼粉饲料(15%)中添加适量水平的苏氨酸, 对提高三倍体虹鳟幼鱼生长、消化酶活性、促进肠道形态结构发育和增强免疫具有积极的作用。     

Effects of dietary soybean meal and photoperiod cycle on osmoregulation following seawater exposure in Atlantic salmon smolts

Atlantic salmon Salmo salar juveniles were fed either fishmeal-based diets (FM) or diets in which soybean meal (SBM) partly replaced the FM from first feeding on. The fish were kept at continuous daylight during the juvenile stage. During the last 3 weeks before reaching 100 g body mass, all fish were subjected to 12L:12D. Starting at 100 g body mass, groups of 60 fish from each feeding background were subjected to continuous light for 12 weeks (short winter), or a square-wave photoperiod cycle to stimulate parr to smolt transformation with 8L:16D during the first 6 weeks, and then continuous light during the last 6 weeks (long winter). After the 12 weeks, 20 fish from each treatment were subjected to 0, 24 or 96 h seawater exposure at a water salinity of 34. Hypo-osmoregulatory ability at seawater exposure was assessed by mortality, intestinal pathology, plasma ion concentrations and osmolality, gill Na⁺/K⁺-ATPase activity and element concentrations in the cytoplasm of distal intestinal enterocytes using X-ray microanalysis. The hypo-osmoregulatory capacity was higher in fish kept at short winter than at long winter, apparently due to more rapid development of gill Na⁺/K⁺-ATPase activity. Fish fed SBM suffered typical soybean meal-induced histological alterations of the distal intestine and apparent reductions in digestive function in the more proximal gastrointestinal regions. The net osmoregulatory capacity of these fish was maintained, as indicated by higher gill Na⁺/K⁺-ATPase activity and lower plasma Na⁺, Ca²⁺ and osmolality compared to the FM-fed fish. Thus, feeding SBM did not impair the hypo-osmoregulatory ability of the Atlantic salmon following seawater exposure.     

大豆低聚糖对牙鲆营养效应的研究：Ⅱ 消化率和消化生理

本试验比较研究了不同蛋白源(鱼粉,FM;大豆分离蛋白,SPI)基础饲料中添加大豆低聚糖(SBOS)对牙鲆(Paralichthys olivaceus)消化道(胃、幽门盲囊、前肠、中肠和后肠)和肝脏消化酶活性、饲料表观消化率和消化道(胃、小肠)组织结构的影响。分别以FM、SPI作为主要蛋白源,配制了4种等氮等能饲料。其中,饲料FM、SPI分别以FM、SPI作为主要蛋白源;饲料FMO、SPIO分别在饲料FM、SPI基础上添加10%SBOS(水苏糖：2.61%;棉籽糖：0.61%)。试验表明：①饲料中添加SBOS普遍降低了牙鲆消化道和肝脏蛋白酶活性,但差异均不显著(p > 0.05)。FM基础饲料中添加SBOS显著降低了肝脏脂肪酶活性(p < 0.05),而对消化道脂肪酶活性无显著影响;SPI基础饲料中添加SBOS显著提高了前肠脂肪酶活性,降低了胃和中肠脂肪酶的活性,而对幽门盲囊、后肠和肝脏脂肪酶活性均无显著影响。FM基础饲料中添加SBOS显著降低了中肠淀粉酶活性,提高了胃淀粉酶活性(p < 0.05),而对幽门盲囊、前肠、后肠和肝脏淀粉酶活性无显著影响;SPI基础饲料中添加SBOS显著提高了胃和前肠淀粉酶活性,降低了中肠脂肪酶活性(p < 0.05),而对幽门盲囊、后肠和肝脏淀粉酶活性无显著影响;②FM基础饲料中添加SBOS显著降低了饲料干物质表观消化率(p < 0.05),而对粗蛋白、粗脂肪和能量表观消化率均无显著影响(p > 0.05)。SPI基础饲料中添加SBOS对干物质、粗蛋白、粗脂肪和能量表观消化率均无显著影响;③饲料中添加SBOS对牙鲆胃和小肠组织结构均无明显负面影响。试验结果表明,饲料中添加SBOS对牙鲆消化道和肝脏消化酶活性、饲料表观消化率和消化道组织结构均没有表现出明显的负面效应;大豆蛋白源中含有的SBOS不是影响牙鲆对其利用率差的主要因素。     

Up-regulation of a non-kinase activity isoform of Ca(2+)/calmodulin-dependent protein kinase kinase beta1 (CaMKKbeta1) in hibernating bat brain

To improve hybrid tilapia (Oreochromis niloticusxO. aureus) survival under cold shock, the influence of diets containing various dietary lipids was investigated. Four different diets were used which consisted of 12% fish oil, 12% palmitoleic oil 12% coconut oil, and a mixture of fish oil (7%) and corn oil (5%). Our results showed that during cold shock, the proportion of saturated fatty acids in the fish steadily and significantly decreased for all of the diets, but the proportion of monounsaturated fatty acids increased. Proportions of polyenoic fatty acids initially increased then stabilized for the mixed, fish, and coconut oil diets, but did not significantly increase until day 4 for the palmitoleic oil diet. The stearoyl-CoA desaturase (SCD) activity was the lowest on day 0 and then gradually increased for all diets. At any point, the enzymatic activity of SCD was the highest for fish on the mixed and the coconut oil diet, followed by the palmitoleic oil diet, and was lowest for the fish oil diet. The expression of SCD mRNA steadily increased for all diets, but increased more substantially for the mixed diet. On day 6, the expression was the highest for fish on the mixed diet, followed by the coconut oil diet, with the lowest levels for those on the palmitoleic and fish oil diets. These results show that dietary lipids strongly affect the fatty acid composition and SCD expression in tilapia under cold shock, and cold tolerance of this species is also affected.     

Dietary nucleotides influence immune responses and intestinal morphology of red drum Sciaenops ocellatus

Dietary nucleotides have been shown to benefit many physiological and nutritional functions in higher vertebrates and fish. Therefore, a 6-week feeding trial was conducted to evaluate the effects of graded levels of a commercial nucleotide product on growth performance, immune responses and intestinal morphology of juvenile red drum (initial average weight of 7.1 g). The basal diet was formulated to contain 40% protein, 10% lipid and a digestible energy level of 3.5 kcal g^?1. Two levels of nucleotide (Ascogen P^®, 0.5% and 1% of diet) were added to the basal diet with menhaden fishmeal and menhaden oil adjusted to provide isonitrogenous and isolipidic diets. Nucleotide supplementation tended to improve weight gain and survival of red drum, but not at a significant level. Neutrophil oxidative radical anion production and serum lysozyme activity tended to be higher for fish fed diets supplemented with nucleotide, while extracellular superoxide anion production of head kidney macrophages from fish fed diets with 1% nucleotide was significantly (P < 0.05) increased, although no significant differences were observed between fish fed 0.5% nucleotide diet and the basal diet.Nucleotide supplementation significantly (P < 0.05) increased fold height in the proximal intestine, and enterocyte height in the pyloric caeca, proximal and distal enteric sections. A significantly (P < 0.05) higher microvilli height was observed in all evaluated enteric sections of fish fed with diets supplemented with nucleotides. It is therefore possible to use dietary nucleotides supplementation to significantly enhance the intestinal structure of red drum. Likewise, nucleotides in the diet may improve some components of the non-specific immune response of this sciaenid fish.     

Intestinal epithelial cell proliferation and migration in Atlantic salmon, Salmo salar L.: effects of temperature and inflammation

A 28-day feeding trial was carried out to characterise intestinal epithelial cell (IEC) turnover in Atlantic salmon (Salmo salar L.) post-smolts in seawater. Four groups of fish raised at two temperatures of 8°C or 12°C and fed two different diets were investigated. The diets included a reference maize gluten and fishmeal-based diet (FM) and an experimental enteropathy-causing diet containing 20% extracted soybean meal (SBM). IEC proliferation and migration were investigated by labelling cells with the in vivo proliferation marker 5-bromo-2′-deoxyuridine (BrdU). Proliferating cell nuclear antigen (PCNA) labelling was used as a control for identifying proliferating cells. Samples of the proximal (PI), mid (MI) and distal (DI) intestinal regions were collected at five time points (3 h–28 days) over the experimental period. Histologically, FM-fed fish had normal mucosa, whereas the SBM-fed fish developed DI enteropathy. Major zones of cell proliferation were observed in the mucosal fold bases for all intestinal regions. Over time, BrdU-labelled cells migrated up mucosal folds to the tips before being lost. Migration rates were dependent on intestinal region, temperature and diet. Highest migration rates were observed in the PI followed by the MI and DI for FM-fed fish. Diet and temperature barely affected migration in the PI and MI. Migration in the DI was most sensitive to diet and temperature, with both SBM and the higher water temperature increasing proliferation and migration rates. The slow IEC turnover in the DI might help to explain the sensitivity of this region to dietary SBM-induced enteropathy.     

Effect of dietary ascorbic acid on growth and non-specific immune responses of tiger puffer, Takifugu rubripes

We report nutritional physiology and non-specific immune responses of ascorbic acid (AA) in puffer fish for the first time. This study aimed to examine the essentiality and requirements of AA in diets for the tiger puffer, Takifugu rubripes based on growth performance, liver AA and bone collagen concentration, and non-specific immune responses. Five casein-gelatin based semi-purified diets were formulated to contain five graded levels of l-ascorbyl-2-monophosphate at 0, 40, 80, 160 and 700mg/kg (designated as AMP0, AMP40, AMP80, AMP160 and AMP700, respectively) and fed to triplicate groups of fish. After 10weeks of feeding trial, growth performances of fish (initial body weight, 35g) fed the AMP0 were significantly lower compared to that of fish fed diets supplemented with AMP. The fish fed the AMP0 diet also exhibited significantly lower hematocrit, condition factor and hepatosomatic index compared to the fish fed diets supplemented with AMP. Phagocytic activity (NBT assay) was significantly lower in fish fed the AMP0 diet than in fish fed the AMP containing diets. Plasma lysozyme activity of fish fed the AMP80 and AMP160 was significantly higher than that of fish fed the AMP0. Dietary supplementation of AMP significantly increased the liver superoxide dismutase in the fish. Myeloperoxidase activity of fish fed the AMP0 was significantly lower compared to that of fish fed the AMP containing diets. Bone collagen level tended to increase numerically and total AA concentration in liver of fish was significantly increased in a dose dependent manner by the supplementation of AMP. Therefore, tiger puffer requires exogenous ascorbic acid and the optimum dietary level could be 29mg AA/kg diet for normal growth and physiology. Dietary AA concentration over 82mg/kg could be required to enhance non-specific immune responses of the fish. However, it does not seem that the fish needs an overdose of dietary AA (>160mg/kg) for better non-specific immune responses.     

Divergent effects of eicosapentaenoic and docosahexaenoic acid ethyl esters, and fish oil on hepatic fatty acid oxidation in the rat

The physiological activity of fish oil, and ethyl esters of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) affecting hepatic fatty acid oxidation was compared in rats. Five groups of rats were fed various experimental diets for 15 days. A group fed a diet containing 9.4% palm oil almost devoid of n-3 fatty acids served as a control. The test diets contained 4% n-3 fatty acids mainly as EPA and DHA in the form of triacylglycerol (9.4% fish oil) or ethyl esters (diets containing 4% EPA ethyl ester, 4% DHA ethyl ester, and 1% EPA plus 3% DHA ethyl esters). The lipid content of diets containing EPA and DHA ethyl esters was adjusted to 9.4% by adding palm oil. The fish oil diet and ethyl ester diets, compared to the control diet containing 9.4% palm oil, increased activity and mRNA levels of hepatic mitochondrial and peroxisomal fatty acid oxidation enzymes, though not 3-hydroxyacyl-CoA dehydrogenase activity. The extent of the increase was, however, much greater with the fish oil than with EPA and DHA ethyl esters. EPA and DHA ethyl esters, compared to the control diet, increased 3-hydroxyacyl-CoA dehydrogenase activity, but fish oil strongly reduced it. It is apparent that EPA and DHA in the form of ethyl esters cannot mimic the physiological activity of fish oil at least in affecting hepatic fatty acid oxidation in rat.     

Effects of a high soy protein diet on intestinal polyamines and ornithine decarboxylase activity in rats

This study was performed to determine whether intestinal luminal polyamine concentrations are affected by a high soy protein diet when compared with a high casein diet or a normoprotein casein diet. We also determined the effects of these diets, with differences in polyamines content, on mucosal polyamines and ornithine decarboxylase (ODC) activity to assess cell proliferation. Three groups of eight male Wistar rats were fed either a 50% soy protein diet, a 50% casein diet, or an 18% casein diet as a control. After 4 weeks of feeding, both intestinal content and mucosa were recovered. Polyamines were assayed by high performance liquid chromatography. ODC activity was measured by the release of (14)CO(2) from (14)C-L-ornithine. Luminal putrescine and cadaverine concentrations were higher in the jejunum than in the ileum, suggesting an absorptive process. The highest concentrations of intestinal polyamines were observed in rats fed the soy protein diet (P < 0.05). Only minor differences were observed in mucosal polyamines according to the diets. ODC activity was also higher in the intestinal mucosa of rats fed the high soy protein diet (P < 0.05). These results suggest that intestinal luminal polyamine concentrations and ODC activity are modulated by the dietary protein source.