Dietary supplementation with chitin and chitosan on haematology and innate immune response in Epinephelus bruneus against Philasterides dicentrarchi

The present study investigated the effect of 1.0% chitin and chitosan supplementation diets on haematology and immune response in Kelp grouper, Epinephelus bruneus against protozoan parasite, Philasterides dicentrarchi. The red blood cells (RBC), white blood cells (WBC), haemoglobin levels, lymphocytes, monocytes, and neutrophils significantly increased in kelp grouper fed with chitin or chitosan enriched diets against P. dicentrarchi. The mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), and thrombocytes did not significantly change against pathogen. The phagocytic activity, respiratory burst activity, complement activity, antiprotease activity, and α2-macroglobulin were significantly enhanced in fish fed with 1% chitin and chitosan diet on weeks 2 and 4. The lysozyme activity, total protein, and myeloperoxidase activity significantly increased in fish fed with chitin or chitosan supplementation diet from weeks 1 to 4 against pathogen. The cumulative mortality was found low in fish fed with chitin and chitosan enriched diets than those of control against pathogen. The present study suggests that supplementation of 1.0% chitin or chitosan in diets positively enhances immune response and affords disease resistance in kelp grouper, E. bruneus against P. dicentrarchi infection.     

Haemato-immunological and growth response of mirror carp (Cyprinus carpio) fed a tropical earthworm meal in experimental diets

An investigation was conducted to evaluate the effect of feeding a tropical earthworm meal (Perionyx escavatus) on the haemato-immunological response and growth performance of mirror carp (Cyprinus carpio). Fish were fed diets for a total of 88 days, fishmeal served as the main protein source in the control diet. Two remaining diets consisted of fishmeal fixed at 33.65% provision of protein and the remaining 66.35% protein was provided by soybean meal (SBM diet) or P. excavatus meal (EW diet). Compared to control and SBM fed fish (7.69 ± 0.28 and 5.92 ± 0.31 g/dl, respectively), a significant increase in haemoglobin was measured in EW fed fish (9.57 ± 0.24 g/dl). Consequently significant elevations were also observed in mean corpuscular haemoglobin (MCH; 79.13 ± 4.59 pg) and mean corpuscular haemoglobin concentration (MCHC; 22.69 ± 0.54 pg) in EW fed fish. On the contrary, compared to control and SBM fed carp total leukocyte levels (2.72 ± 0.17 and 3.10 ± 0.17 × 10(4)/mm(3), respectively) were significantly decreased in the EW group (2.15 ± 0.14 × 10(4)/mm(3)). Moreover at day 14 and 21 post immunisation with bacterin isolated from Aeromonas hydrophila fish fed the EW diet displayed a significant reduction in respiratory burst activity (RBA) compared to control and SBM fed fish. After 60 days of feeding, fish fed EW diet showed a significant elevation in final body weight compared to fish fed a fishmeal based diet (control treatment) and fish fed a soybean meal based diet. Similar improvements were observed in feed utilisation efficiency. The present study shows that feeding P. excavatus meal to mirror carp decreases some aspects of the innate immune response, but at the same time gives rise to significant enhancement of growth and feed utilisation efficiency.     

大黄鱼稚鱼L-蛋氨酸需要量的研究

研究了1242日龄大黄鱼（Pseudosciaena crocea R.）稚鱼蛋氨酸需要量。以白鱼粉、磷虾粉和乌贼粉作蛋白源,通过添加L-晶体氨基酸使饲料与大黄鱼卵的必需氨基酸组成一致（蛋氨酸除外）,制成6种等氮（8.8%）等能（16.65 kJ/g）的微黏合饲料。L-蛋氨酸梯度依次为饲料的1.19%、1.62%、2.18%、2.65%、3.13%和3.66%,或饲料蛋白的2.17%、2.95%、3.95%、4.81%、5.70%和6.65%。以生物饵料（丰年虫无节幼体和桡足类）作对照组。每处理设3个重复,每桶（180 L）内随机放3500尾初始体重为（1.930.11） mg 的12日龄大黄鱼稚鱼。实验为期30d。结果显示,稚鱼的成活率随饲料蛋氨酸水平的升高而升高,在2.18%蛋氨酸水平时达到最高,之后无显著变化。特定生长率（SGR）随饲料蛋氨酸水平的升高而升高,在2.18%蛋氨酸水平时达到最高,之后则呈下降趋势。对照组稚鱼的成活率和SGR最高,均显著高于蛋氨酸组（P0.05）。各组间稚鱼体脂肪和灰分差异不显著,但体蛋白随饲料蛋氨酸水平的升高而升高,在2.65%蛋氨酸水平时达到最高,之后则稍微下降;对照组鱼体蛋白和各必需氨基酸含量均显著高于其他组（P0.05）。经二次多项式模型分析,1242日龄大黄鱼稚鱼的蛋氨酸需要量为饲料的2.58%或饲料蛋白的4.69%。       

Nutritional properties of four marine microalgae for albino rats

The nutritive value of the marine microalgaeTetraselmis suecica, Isochrysis galbana, Dunaliella tertiolecta andChlorella stigmatophora was studied in diets given to rats. Biological assays were carried out in order to determine the Protein Efficiency Ratio (PER) and the Food Conversion Efficiency (FCE). Each dried microalga was fed to weaning Wistar albino rats as the sole protein source at a protein level of 12%. Control rats were given diets containing 12 % casein. Food consumption was similar in all groups. PER values obtained were 1.14 withT. suecica diet, 1.13 withI. galbana diet, 2.07 withD. tertiolecta diet and 1.13 withC. stigmatophora diet (casein, 2.50). FCE values followed a similar pattern. The data showed that the marine microalgaD. tertiolecta is a source of protein of good quality. Its PER is quite high, compared to vegetable and cereal proteins, and compares favourably with other microbial protein sources, such as yeasts or different freshwater microalgae. Haematological tests showed no significant differences among the groups in haemoglobin levels, red and white blood cell counts, differential count and mean corpuscular volume. Different blood parameters were also determined and a significant decrease in triglycerides levels appeared with all the microalgal diets, whereas a significant decrease in cholesterol appeared inD. tertiolecta andC. stigmatophora diets.     

Supplementation of orotic acid to the casein, but not to egg protein, soy protein, or wheat gluten diets, increases serum ornithine carbamoyltransferase activity

Effects of dietary supplementation of orotic acid to a diet containing the casein protein were compared with diets containing egg protein, soy protein, or wheat gluten on lipid levels in the liver and serum and activities of ornithine carbamoyltransferase (OCT) and alanine aminotransferase in the serum of rats. We found that supplementation of orotic acid to each diet increased the contents of the liver total lipids, triacylglycerol, and phospholipids compared with those not supplemented. The contents of liver total lipids, triacylglycerol, cholesterol, and phospholipids in rats fed the casein diet were significantly higher than those of rats fed the other three diets when orotic acid was supplemented. The levels of triacylglycerol, cholesterol, and phospholipids in the serum of rats fed the casein diet were markedly decreased by addition of orotic acid. The supplementation of orotic acid significantly increased the activities of both serum OCT and alanine aminotransferase in rats fed the casein diet, but not in rats fed the other diets. In conclusion, liver lipid accumulation induced by dietary orotic acid depends on the type of dietary protein. The enhancement of serum OCT activity may result from liver lipid accumulation in rats fed the casein diet supplemented with orotic acid, demonstrating hepatic damage.     

Depletion of delta 9 desaturase (EC 1.14.99.5) activity in lactating rat during protein restriction

The effects of protein restriction on the activity of delta9 desaturase (EC 1.14.99.5) were investigated in lactating rats. A control group was fed a balanced diet (20% casein) for 14 days, whereas the experimental groups were fed a low-protein diet (8% casein), supplemented with or without L-methionine (0.4%), for 14 days. The enzyme activity was measured by incubations of hepatic microsomal pellets with (1-14C) stearic acid. Results showed a decreased delta9 desaturase activity, after 2,7 and 14 days of depleted diet, of -50, -40 and -33% respectively, compared with control. The supplementation of the low-protein diet with 0.4% methionine, which favours food consumption as well as growth, did not improve the altered delta9 desaturase activity. Our data evidenced that delta9 desaturase activity is depleted by protein restriction during lactation, when the protein needs are high for the biosynthesis of animal tissues. This change has to be considered as a sign of depressed delta9 desaturase biosynthesis or modifications of enzymatic properties, or both.     

Toxic effects of fermented and unfermented sorghum meal diets naturally contaminated with mycotoxins

Male weanling rats fed diets containing fermented and unfermented tannin-free sorghum meals naturally contaminated with traces of ochratoxin A, zearalenone, and an unidentified toxic substance developed anorexia. Mean changes in body weight over a 28-day feeding period for rats fed fermented and unfermented sorghum meal and ANCR casein diets of 8% protein were -2.8, +13.8, and +100.5 g, respectively. Rats fed fermented sorghum meal developed alopecia; hematological findings showed a decreased mean corpuscular volume, hypochromic microcytosis, balanced leukopenia, and hypoproteinuria . Histological findings showed testicular hypoplasia of the germinal epithelium and no mature spermatozoa. Necrosis and mineralization in Henle's loop were also observed. No damage was apparent to the liver, cerebellum, cerebrum, spleen, or adrenal glands.     

Toxic effects of fermented and unfermented sorghum meal diets naturally contaminated with mycotoxins.

Male weanling rats fed diets containing fermented and unfermented tannin-free sorghum meals naturally contaminated with traces of ochratoxin A, zearalenone, and an unidentified toxic substance developed anorexia. Mean changes in body weight over a 28-day feeding period for rats fed fermented and unfermented sorghum meal and ANCR casein diets of 8% protein were -2.8, +13.8, and +100.5 g, respectively. Rats fed fermented sorghum meal developed alopecia; hematological findings showed a decreased mean corpuscular volume, hypochromic microcytosis, balanced leukopenia, and hypoproteinuria . Histological findings showed testicular hypoplasia of the germinal epithelium and no mature spermatozoa. Necrosis and mineralization in Henle's loop were also observed. No damage was apparent to the liver, cerebellum, cerebrum, spleen, or adrenal glands.     

Effects of alginates and sodium carrageenates mixed with soy proteins on the coefficient of protein efficiency

Male 21 d-old Wistar rats, were fed for 4 wk with diets containing casein or soybean proteins (10%) with 0.5, 1, 2 or 3% sodium alginate or sodium carrageenan or without any alginate or carrageenan. Daily protein intake and weight gain of casein-fed rats were not significantly different (P less than 0.05) from those of rats fed soybean meal with alginate, whatever the dose received. Rats fed 3% carrageenan in soybean meal had significantly higher feed intake than that of rats fed casein. At the levels studied, alginate had no effect on the Protein Efficiency Ratio (PER), but carrageenan did. The addition of increased quantities of carrageenan to soybean meal followed by heating the mixture led to a progressive and significant decrease in PER at all levels of carrageenan compared to casein feeding. The addition of 3% carrageenan to heated soybean meal, corresponding to 0.62% of meal diet, led to a significant decrease in PER. These results confirm the precipitating role of carrageenans on proteins.     

Cell growth of skeletal muscle as an indicator of protein requirements of the adult rat

Skeletal muscle growth, muscle nucleic acids and muscle protein synthesis capacity, were measured to evaluate the protein requirement of adult rats. Wistar rats were fed on diets containing 4%, 10% or 20% casein + D,L-methionine. All diets were provided for 21 days beginning at 90 days of age. Body weight, food efficiency and net weight change increased as the casein content of the diet increased. Muscle DNA, RNA and RNA/protein were lost, but protein and protein/DNA increased on the 4% and 20% protein diet. This fact involves an aplasia phenomenon although the hypertrophic growth is maintained. Alterations of the insulin and GH plasma levels were observed. These findings indicate that for adult rats the 4% and 20% protein diets are not adequate for the period of adult maintenance.     

High protein diet in pregnancy. Effects on neonatal metabolism

The effect of a high protein diet (20% casein + D,L-methionine) administered to Wistar rats during pregnancy on some aspects of cellular growth and RNA metabolism of progeny has been studied. Comparisons were made with well-nourished (10% casein + D,L-methionine) controls. Newborns individual weight, litter weight and number of newborns per litter were unmodified. However, neonate protein content dropped significantly when compared with controls. Both rate of DNA and number of nuclei were unchanged. Protein/DNA ratio (cellular size relative to protein) decreased, which might have led to an atrophy phenomenon, even if the newborn weight/number of nuclei ratio was not modified. Acid DNase activity rose, bringing about DNA breakdown. Total RNA content together with RNase activity fell in newborn from rats suffering high protein diet. Moreover, protein synthesis capacity (RNA/protein ratio) did not change. These results suggest that the administration of a high protein diet to pregnant rats lead to changes in newborn protein rate and nucleic acid turnover by modulating specific nuclease activity.     

Effect of Glutathione on Lipogenesis in Liver Slices from Rats Fed on Low Casein Diet

To study the mechanism of fatty infiltration in the liver due to added sulfur-containing amino acids to low casein diet, the effect of sulfur-containing amino acids and glutathione (GSH) on the incorporation of acetate-l-¹⁴C into lipid fractions were studied in liver slices from rats fed on 8% casein diet (Basal diet) with or without added methionine (Met).

The liver acetyl Co A carboxylase activities of rats on basal diet with or without added Met were similar.

Addition of Met, cystine or cysteine to the incubation medium had little effect on lipogenesis of slices. On addition of GSH to liver slices from rats fed on basal diet, lipid formation increased appreciably. On the other hand, addition of GSH to liver slices from rats fed on Met supplemented diet showed no accelerative effect on lipogenesis.

Addition of GSH to the incubation medium of liver slices from rats fed on basal diet tended to reduce the incorporation of acetate into the phospholipid fraction and to increase into the fatty acid fraction of liver slices.

The content of liver GSH was lower in rats on basal diet than in those on Met supplemented diet. The higher GSH level in rats on Met supplemented diet may be one factor causing fatty infiltration in the liver of these animals.     

Effects of amino acid supplementation on the nutritive quality of fermented linseed meal protein in the diets for rohu, Labeo rohita, fingerlings

A feeding trial was conducted for 8 weeks to examine the effects of partial substitution of fish meal (FM) protein (crude protein content: 58.5%) with linseed meal protein with and without supplemental amino acids in diets for rohu Labeo rohita (Hamilton), fingerlings (mean weight: 1.50 ± 0.3 g). Prior to incorporation into the diets, linseed meal was fermented with lactic acid bacteria ( Lactobacillus acidophilus ) to reduce/eliminate the antinutritional tannin and phytic acid factors. Twelve experimental diets (diets D1–D12) were formulated to replace the FM protein from a reference diet (RD) with linseed meal protein at different levels (four sets of diets, of which each set of three diets contained 25%, 50% and 75% replacement of FM protein by linseed meal protein, respectively). Diets D1–D3 were not supplemented with any amino acid. Lysine was supplemented in diets D4–D6. Diets D7–D9 were supplemented with methionine + cystine (together), and diets D10–D12 contained lysine and methionine + cystine (together). Lysine and methionine + cystine (together) were added to the diets at 5.7% and 3.1% of dietary protein, respectively. The groups of fish fed diets without amino acid supplementation had significantly lower percentages of weight gain, specific growth rate and high feed : gain ratio than the fish groups fed other experimental diets. The addition of lysine and methionine + cystine to the diet in which 50% of the FM protein was replaced by linseed meal protein (diet D11) significantly improved fish performance. The results of the present study suggest that rohu fingerlings can effectively utilize the supplemented amino acids and that linseed meal protein can replace up to 50% of the FM protein in rohu diets if the linseed meal is properly processed (fermented) and supplemented with the lacking amino acids.     

Relationship between dietary tyrosine and plasma cholesterol in the rat

The present study was undertaken to measure the effects of dietary tyrosine added to fish protein and peanut meal on plasma cholesterol and plasma thyroid hormone levels in the rat. These dietary proteins were chosen because they contain similar amounts of tyrosine but release it at different rates during enzymatic hydrolysis. Casein was chosen as the reference protein. Supplementation was used to obtain tyrosine levels similar to that of casein. Male Sprague-Dawley rats were fed cholesterol-enriched diets containing 15% protein. After 3 weeks of experimental feeding, total postprandial plasma cholesterol was similar in the casein and peanut meal groups and significantly lower in the fish group. When added to the fish diet, tyrosine caused an increase in plasma cholesterol to a level similar to that of the casein group, whereas supplementation had no effect on plasma cholesterol of rats fed the peanut meal diet. The effects of dietary proteins or of tyrosine supplementation on cholesterol levels of the (density less than 1.006 g/mL) lipoprotein fraction were comparable, but not all significant, to those observed on total plasma cholesterol. In addition, casein and fish diets induced significantly higher levels of plasma triiodothyronine (T3) and lower levels of plasma thyroxine (T4) than did the peanut meal diet. However, the addition of tyrosine to the fish or the peanut meal diet did not modify the plasma thyroid hormone levels.(ABSTRACT TRUNCATED AT 250 WORDS)     

Lunatoic Acid A,a Morphogenic Substance Inducing Chlamydospore-like Cells in Some Fungi

The effect of the addition of 0.26 % free tryptophan (Trp) to a 20 % casein diet containing 6 mg of nicotinic acid per 100 g of diet on the ratio of N¹-methyl-2-pyridone-5-carboxamide (2-py) plus N¹-methyl-4-pyridone-3-carboxamide (4-py) to -methylnicotinamide (MNA) excretion was investigated in rats. The urinary excretion of MNA, 2-py and 4-py, respectively, increased statistically significantly with the feeding of a 0.26% Trp (the same as the content of the 20% casein diet) supplemented 20% casein diet, although it did not increase with the feeding of a 40% casein diet, compared with in the case of the 20 % casein diet [Agric. Biol. Chem., 52, 1765 (1988)]. So, the total urinary excretion of Nam and its metabolites was 1.8 times higher in the group fed the Trp supplemented diet than in the group fed the 20 % casein diet. However, the ratio of 2-py plus 4-py to MNA excretion was much lower in the group fed the Trp supplemented diet than in the group fed the 20 % casein diet (13.16 ± 3.75→5.49 ± 2.25). This decreased ratio was considered to be partially due to a decrease in the 4-py forming MNA oxidase, which decreased significantly with the feeding of the Trp supplemented diet. Furthermore, the metabolic fate of Trp was greatly affected by the form of Trp, free or bound.     

Effect of high fluoride intake on haematological aspects of the mouse.

The effect of feeding adult Swiss albino mice of both sexes a diet supplemented with 0, 125, 250 and 500 parts/10(6) of fluoride for four and eight week periods on haemoglobin concentration (Hb), packed cell volume (PCV) and mean corpuscular haemoglobin concentration (MCHC) was investigated. Values of the three parameters were significantly lowered at both periods in the treated groups as compared with the controls. The extent of reduction in these values was, in general, dependent on the dose of supplemented dietary fluoride. Clinical symptoms were not observed before the end of the sixth week. However, appearance of the symptoms did not change the trend of variations in Hb, PCV and MCHC values. The reduced values could be the result of lowered haemoglobin synthesis and erythropoiesis. It was suggested that these haematological indices could serve to detect preclinical effects of high fluoride intake with an added dose of as low as 125 parts/10(6), or even less, for a period of four weeks or probably earlier.     

Kalopanax pictus as feed additive controls bacterial and parasitic infections in kelp grouper, Epinephelus bruneus

Feeding kelp grouper, Epinephelus bruneus (26.1 ± 1.4), with 0%, 0.1%, 1.0%, and 2.0% Kalopanax pictus extract-supplementation diets, for 30 days reduced mortality on being challenged intraperitoneally 100 μl with bacterium Vibrio alginolyticus (2.1 × 10(7) cfu ml(-1)) and ciliate parasite Philasterides dicentrarchi (2.3 × 10(7) ciliates ml(-1)). The red blood cells (RBC), white blood cells (WBC), haemoglobin, haematocrit, lymphocytes, and monocytes levels significantly increased in kelp grouper fed with all doses of K. pictus-supplementation diets and challenged with bacterium and parasite when compared to control. However, the levels of mean corpuscular volume (MCV), mean corpuscular haemoglobin (MCH), mean corpuscular haemoglobin concentration (MCHC), neutrophils, and thrombocytes did not significantly. The phagocytic activity, complement activity, and antiprotease activity did not significantly change in kelp grouper fed with 0.1% K. pictus-supplementation diets and challenged with bacterium and parasite. The respiratory activity, lysozyme activity, bactericidal activity, total protein level, and myeloperoxidase levels significantly increased in kelp grouper fed with all the doses of K. pictus-supplementation diet and challenged with bacterium and parasite. However, α2-macroglobulin level significantly increased with 1.0% diet, but not with 0.1% and 2.0% diets. Therefore this study suggests that 1.0% and 2.0% K. pictus-supplementation diets positively protected and enhanced the immune system in kelp grouper E. bruneus against V. alginolyticus and P. dicentrarchi infection.     

Effects of Fermentation Conditions and Aging Temperature on Volatile Ester Contents in Wine

In these present experiments, rats were fed a low casein, vitamin E and selenium deficient diet and were killed at various time intervals up to the occurrence of massive liver necrosis. Liver malondialdehyde, and liver and serum vitamin E levels were analyzed simultaneously with a pathological investigation of the features of the liver and measurement of serum GOT and GPT activities. These investigations were also performed on control rats given a high casein, vitamin E defincient diet or a low casein, vitamin E supplemented diet.

The results show that the liver malondialdehyde level was not increased concomitant with the decrease in liver and serum vitamin E level nor with the occurrence of massive liver necrosis. Liver cells of the rats fed on the low casein, vitamin E deficient diet showed swelling of the cytoplasm at the initial stage, and progressive centrilobular lipid deposition was observed by Sudan III stain.     

Potential use of high levels of vegetal proteins in diets for market-sized gilthead sea bream (Sparus aurata)

The effect of partial or total dietary substitution of fishmeal (FM) by vegetal protein sources on growth and feed efficiency was carried out in on-growing gilthead sea bream (mean initial weight 131 g). The Control diet (FM 100) contained FM as the primary protein source, while in Diets FM 25 and FM 0 the FM protein was replaced at 75% and 100%, respectively, by a vegetable protein mixture consisting of wheat gluten, soybean meal, rapeseed meal and crystalline amino acids. Diets FM 25 and FM 0 also contained krill meal at 47 g/kg in order to improve palatability. At the end of the trial (after 158 d), fish survival was above 90%. Final weight and the specific growth rate were statistically lower in fish fed the Control diet (361 g and 0.64%/d), compared with 390–396 g and 0.69–0.70%/d after feeding vegetal diets. No significant differences were found regarding feed intake and feed conversion ratio. The digestibility of protein and amino acids (determined with chromium oxide as indicator) was similar in all diets. The blood parameters were not significantly affected by treatments. The activity of trypsin and pepsin was significantly reduced after feeding Diet FM 0. In the distal intestine, the villi length in fish fed Diet FM 25 was significantly longer and the intestine of the fish fed the FM 100 diet showed a smaller number of goblet cells. In conclusion, a total FM substitution by a vegetal mix supplemented with synthetic amino acids in on-growing sea bream is feasible.     

Effects of dietary oyster extract on lipid metabolism, blood pressure, and blood glucose in SD rats, hypertensive rats, and diabetic rats

Oyster extract was prepared by hydrolysis of oyster protein with proteases, Aloase (a protease from Bacillus subtilis), and Pancitase (a protease from Aspergillus oryzae). Rats were fed a diet containing 20% casein (the control diet) or 15% casein and 5% oyster extract (the oyster extract diet) as the protein source. The oyster extract diet exerted a significant reduction in serum cholesterol and liver triglyceride concentrations as compared with the control diet in Sprague-Dawley (SD) rats fed cholesterol-supplemented diets for 4 weeks. The activities of cytosolic fatty acid synthase and glucose-6-phosphate dehydrogenase were significantly lower in the oyster extract group than in the control group in the liver of SD rats. Hepatic cholesterol and triglyceride concentrations were significantly lower in spontaneously hypertensive (SH) rats and Otsuka Long-Evans Tokushima Fatty (OLETF) rats, type 2 diabetic rats, fed the oyster extract diet, for 4 weeks and 4 months respectively, than in those fed the control diet in the cholesterol-free diet. Blood pressure was significantly lower in the oyster extract group than in the control group at the 2nd and 4th weeks after the beginning of feeding experimental diets in SH rats. These results suggest that oyster extract prepared by hydrolysis of oyster induces triglyceride-lowering activity in the liver through a decrease in hepatic lipogenesis in SD rats, and that it exerts the antihypertensive effect in SH rats.