Increased Plasma Homocysteine Concentration in Rats from a Low Casein Diet

Rats were fed on a 10% casein (10C) diet, 30% casein (30C) diet, 10C+0.5% methionine diet, or 30C+0.5% methionine diet for 14 d to investigate the relationship between the dietary protein level and plasma homocysteine concentration. The plasma homocysteine concentration was significantly higher in the rats fed on the 10C diet than in the rats fed on the 30C diet, and this phenomenon persisted even under the condition of methionine supplementation. The activity of hepatic cystathionine β-synthase (CBS) was significantly lower in the rats fed on the 10% casein diets than in the rats fed on the 30% casein diets, irrespective of methionine supplementation. This is the first demonstration of a low-protein diet increasing the plasma homocysteine concentration in experimental animals. It is suggested that the decreased CBS activity might be associated, at least in part, with the hyperhomocysteinemia caused by the low-casein diet.     

Increased plasma homocysteine concentration in rats from a low casein diet

Rats were fed on a 10% casein (10C) diet, 30% casein (30C) diet, 10C+0.5% methionine diet, or 30C+0.5% methionine diet for 14 d to investigate the relationship between the dietary protein level and plasma homocysteine concentration. The plasma homocysteine concentration was significantly higher in the rats fed on the 10C diet than in the rats fed on the 30C diet, and this phenomenon persisted even under the condition of methionine supplementation. The activity of hepatic cystathionine beta-synthase (CBS) was significantly lower in the rats fed on the 10% casein diets than in the rats fed on the 30% casein diets, irrespective of methionine supplementation. This is the first demonstration of a low-protein diet increasing the plasma homocysteine concentration in experimental animals. It is suggested that the decreased CBS activity might be associated, at least in part, with the hyperhomocysteinemia caused by the low-casein diet.     

Effects of supplementing a CP-reduced diet with rumen-protected methionine on Fleckvieh bull fattening

The objective of this study was to evaluate the effect of supplementing a CP-reduced diet with rumen-protected methionine on growth performance of Fleckvieh bulls. A total of 69 bulls (367 ± 25 kg BW) were assigned to three feeding groups (n = 23 per group). The control (CON) diet contained 13.7% CP and 2.11 g methionine/kg diet (both DM basis) and was set as positive control. The diet reduced in CP (nitrogen) (RED) diet as negative control and the experimental RED + rumen-protected methionine (MET) diet were characterised by deficient CP concentrations (both 9.04% CP). The RED + MET diet differed from the RED diet in methionine concentration (2.54 g/kg DM vs. 1.56 g/kg DM, respectively) due to supplementation of rumen-protected methionine. Rumen-protected lysine was added to both RED and RED + MET at 2.7 g/kg DM to ensure a sufficient lysine supply relative to total and metabolisable protein intake. Metabolisable energy (ME) and nutrient composition were similar for CON, RED, and RED + MET. Bulls were fed for 105 days (d) on average. Individual feed intake was recorded daily; individual BW was recorded at the beginning of the experiment, once per month, and directly before slaughter. At slaughter, blood samples were collected and carcass traits were assessed. Reduction in dietary CP concentration reduced feed intake, and in combination with lower dietary CP concentration, daily intake of CP for RED and RED + MET was lower compared with CON (P < 0.01). Daily ME intake was reduced in RED and RED + MET compared with CON (P < 0.01). Consequently growth performance and carcass weights were reduced (both P < 0.01) in both RED and RED + MET compared with CON. Supplemental rumen-protected methionine was reflected in increased serum methionine concentration in RED + MET (P < 0.05) as compared to RED but it did not affect growth performance, carcass traits and serum amino acid (AA) concentrations, except for lysine which was reduced (P < 0.01) compared to CON and RED. In conclusion, bulls fed RED or RED + MET diets were exposed to a ruminal CP deficit and subsequently a deficit of prececal digestible protein, but methionine did not appear to be the first-limiting essential AA for growth under the respective experimental conditions.     

Maximal growth occurs at a broad range of essential amino acids to total nitrogen ratios in kittens

Summary Kittens fed diets containing 2.0 and 3.0 times (X) the NRC (1986) essential amino acid (EAA) requirement (EAArq) and 210 to 560g crude protein (CP)/kg diet had growth rates and plasma amino acid patterns that were not significantly different than kittens fed a control diet (CD) containing 1.5 X EAArq and 350 g CP/kg diet. Growth rates of kittens fed diets containing only EAA (with nontoxic levels of arginine and methionine) and 280 to 460 g CP/kg diet were equivalent to those of kittens fed CD. Kittens fed only EAA and 140 and 210 g CP/kg diet had growth rates that were significantly lower than kittens fed CD. Since the growth rate of kittens fed 1.5X EAArq and 210 g CP/kg diet in a previous experiment was equivalent to kittens fed CD (Taylor et al., 1997), it is suggested that the requirement for CP is higher (up to 280 g CP/kg diet) when only EAA are fed. The higher crude protein requirement appears to be primarily a consequence of the high obligatory nitrogen loss as urea (especially from arginine) incurred in the conversion of nitrogen from EAA to dispensable amino acids in the liver and secondarily because of a slow rate of catabolism of the EAA. A 3-dimensional plot of weight gains vs. CP levels and EAA to total nitrogen (E: T) ratios of kittens shows a broad range of CP levels and E: T ratios that support optimal growth in the kitten. It is suggested that similar patterns would occur in the chick, rat and other species if adverse effects caused by excesses of specific amino acids are avoided.Presented in part at the Biology '96 Meeting in Washington, DC [Taylor TP, Deberry J, Morris JG, Rogers QR (1996) Effects of dietary nitrogen level and essential amino acid: total nitrogen (E: T) ratio on kitten weight gain. FASEB J 10: A737 (abs. 4259)] and in part at the Proceedings of the Waltham International Symposium, Pet Nutrition and Health in the 21st Century, Orlando, Florida, USA [Rogers QR, Taylor TP, Morris JG (1997) Optimizing dietary amino acid patterns at various levels of crude protein for the cat. J Nutr (abs.) (in press).     

Reevaluation of dietary methionine requirement by plasma methionine and ammonia concentrations in surgically modified rainbow trout,Oncorhynchus mykiss

This study aimed to reevaluate the dietary methionine requirement by means of the plasma methionine and ammonia concentrations in surgically modified rainbow trout, Oncorhynchus mykiss. A total of 35 rainbow trout averaging 505 ± 6.5 g (initial body weight, mean ± SD) were randomly distributed into seven groups with five fish in each group. After 48 h of food deprivation, each group was fed one of seven L‐amino acid‐based diets containing graded levels of methionine (0.25, 0.40, 0.50, 0.60, 0.70, 0.80 or 0.95% of diet, dry matter bases) by intubation at 1% bodyweight. Blood samples were taken at 0, 5 and 24 h after intubation. Post‐prandial plasma free methionine concentrations (PPmet, 5 h after intubation) and post‐absorptive plasma free methionine concentrations (PAmet, 24 h after intubation) of fish fed diets containing 0.60% or more methionine were significantly (P < 0.05) higher than those of fish fed diets containing 0.50% or less methionine. PPmet and PAmet in fish fed diets containing 0.60% or higher methionine were not significantly different except the PPmet of fish fed a diet containing 0.95% methionine. Post‐prandial plasma ammonia concentrations (PPA, 5 h after intubation) of fish fed diets containing 0.70% or more methionine were significantly higher than those of fish fed diets containing 0.60% or less methionine, and PPA of fish fed diets containing 0.25% and up to 0.60% methionine were not significantly different from each other. Broken‐line model analyses on PPmet, PAmet, and PPA indicated that the dietary methionine requirement of rainbow trout was between 0.59% (1.69) and 0.67% (1.91) of diets (% dietary protein bases) when the diets contained 0.5% cystine.     

有氧运动对高蛋氨酸饮食大鼠血浆NO、ET和NO/ET系统的影响

目的：探讨有氧运动对高蛋氨酸饮食大鼠血浆总一氧化氮合成酶（T-NOS）、一氧化氮（NO）、内皮素（ET）和NO/ET系统的影响。方法：雄性Wistar大鼠随机分为正常饮食对照组（对照组）、高蛋氨酸饲料组（高蛋氨酸组）和有氧运动＋高蛋氨酸饮食组（运动干预组）。对照组喂饲普通饲料,高蛋氨酸组和运动干预组喂饲含3%蛋氨酸的高蛋氨酸饲料,运动干预组同时每日同时进行90 min无负重游泳运动,实验共8周。分别测定血浆同型半胱氨酸（Hcy）、ET、NO和T-NOS含量。结果：高蛋氨酸组血浆Hcy含量显著高于对照组达2倍以上（P〈0.01）,T-NOS和NO含量显著降低,ET含量显著升高（P〈0.01）,且NO/ET比值均显著降低（P〈0.05）;与高蛋氨酸组相比,运动干预组血浆Hcy含量显著下降（P〈0.05）,T-NOS,NO含量和NO/ET比值显著升高（P〈0.05）,且与对照组相比上述各项指标无显著差异。结论：高蛋氨酸饮食可诱发大鼠高同型半胱氨酸血症,血浆NO/ET失衡;有氧运动可降低高蛋氨酸饮食大鼠血浆Hcy水平,改善NO/ET失衡,预防高同型半胱氨酸血症。     

Effects of methyl-deficient diets on methionine and homocysteine metabolism in the pregnant rat

Although the importance of methyl metabolism in fetal development is well recognized, there is limited information on the dynamics of methionine flow through maternal and fetal tissues and on how this is related to circulating total homocysteine concentrations. Rates of homocysteine remethylation in maternal and fetal tissues on days 11, 19, and 21 of gestation were measured in pregnant rats fed diets with limiting or surplus amounts of folic acid and choline at two levels of methionine and then infused with L-[1-(13)C,(2)H(3)-methyl]methionine. The rate of homocysteine remethylation was highest in maternal liver and declined as gestation progressed. Diets deficient in folic acid and choline reduced the production of methionine from homocysteine in maternal liver only in the animals fed a methionine-limited diet. Throughout gestation, the pancreas exported homocysteine for methylation within other tissues. Little or no methionine cycle activity was detected in the placenta at days 19 and 21 of gestation, but, during this period, fetal tissues, especially the liver, synthesized methionine from homocysteine. Greater enrichment of homocysteine in maternal plasma than placenta, even in animals fed the most-deficient diets, shows that the placenta did not contribute homocysteine to maternal plasma. Methionine synthesis from homocysteine in fetal tissues was maintained or increased when the dams were fed folate- and choline-deficient methionine-restricted diets. This study shows that methyl-deficient diets decrease the remethylation of homocysteine within maternal tissues but that these rates are protected to some extent within fetal tissues.     

Dietary methionine effects on plasma homocysteine and HDL metabolism in mice

The effects of dietary manipulation of folate and methionine on plasma homocysteine (Hcy) and high-density lipoprotein cholesterol (HDL-C) levels in wild-type and apolipoprotein-E-deficient mice were determined. A low-folate diet with or without folate and/or methionine supplementation in drinking water was administered for 7 weeks. Fasted Hcy rose to 23 microM on a low-folate/high-methionine diet, but high folate ameliorated the effect of high methionine on fasted plasma Hcy to approximately 10 microM. Determination of nonfasted plasma Hcy levels at 6-h intervals revealed a large diurnal variation in Hcy consistent with a nocturnal lifestyle. The daily average of nonfasted Hcy levels was higher than fasted values for high-methionine diets but lower than fasted values for low-methionine diets. An acute methionine load by gavage of fasted mice increased plasma Hcy 2.5 h later, but mice that had been on high-methionine diets had a lower fold induction. Mice fed high-methionine diets weighed less than mice fed low-methionine diets. Based on these results, two solid-food diets were developed: one containing 2% added methionine and the other containing 2% added glycine. The methionine diet led to fasted plasma Hcy levels of >60 microM, higher than those with methionine supplementation in drinking water. Mice on methionine diets had >20% decreased body weights and decreased HDL-C levels. An HDL turnover study demonstrated that the HDL-C production rate was significantly reduced in mice fed the methionine diet.     

Modification of lead toxicity and organ distribution by dietary sulfur amino acids in chicks (Gallus domesticus)

Chicks (Gallus domesticus) were fed a basal diet deficient in methionine and total sulfur amino acids with 0 or 1000 ppm added lead for 21 days. Methionine or methionine and cystine addition improved growth regardless of dietary lead level. Cystine addition alone improved growth only when lead was present. Relative inhibition of growth by lead was greater with diets containing no added methionine. Hepatic non-protein sulfhydryl concentration was increased by lead and all amino acid additions. Organ lead concentrations were generally lower with added amino acids. Dietary methionine appears to counteract lead toxicity more effectively than cystine.     

Cysteine supplementation decreases plasma homocysteine concentration in rats fed on a low-casein diet in rats

The effect of dietary supplementation with cysteine on the plasma homocysteine concentration was investigated in rats fed on 10% casein (10C) and 30% casein (30C) diets. The 10C diet significantly increased the plasma homocysteine concentration as compared with the 30C diet. The hyperhomocysteinemia induced by the 10C diet was significantly suppressed by cysteine supplementation even at a 0.3% level, whereas cysteine did not decrease the plasma homocysteine concentration when added to the 30C diet. In contrast, 0.3% methionine supplementation of the 10C diet tended to increase the plasma homocysteine concentration. Cysteine supplementation to rats fed on the 10C diet did not alter the plasma cysteine concentration and the hepatic activities of cystathionine beta-synthase and betaine:homocysteine S-methyltransferase, whereas it significantly decreased the hepatic concentrations of S-adenosylmethionine and betaine. These results suggest that cysteine supplementation might be effective for suppressing the hyperhomocysteinemia induced by a low-protein diet.     

鸡肉粉完全替代鱼粉饲料中补充包被氨基酸对凡纳滨对虾生长、体成分及组织氨基酸含量的影响

为了探讨鸡肉粉完全替代鱼粉时饲料氨基酸的平衡性以及外源氨基酸的添加方式与凡纳滨对虾生长、体成分、血浆游离氨基酸及肌肉氨基酸含量的关系, 本试验采用26因子试验设计进行了为期56d的饲养试验。2个饲料蛋白质水平分别为40%和32%, 6个饲料处理分别为鱼粉组(对照组)、鸡肉粉组、鸡肉粉+晶体EAA组、鸡肉粉+晶体EAA+晶体NEAA组、鸡肉粉+包被EAA组、鸡肉粉+包被EAA+包被NEAA组, 配制12组饲料。将凡纳滨对虾(0.300.01) g随机分配到36个圆桶(150 L)中, 每桶30尾, 每3个桶为一个处理组, 饲喂一种饲料, 每天饱食投喂三次。在每一饲料蛋白质水平下, 无论是补充晶体氨基酸(CAA)组还是包被氨基酸组对虾的增重率均显著高于鸡肉粉组(P0.05), 且在32%蛋白质水平下, 包被EAA组对虾增重率达到了鱼粉组水平(P0.05); 补充晶体EAA+NEAA组对虾增重率与补充晶体EAA组无差异(P0.05), 但均显著低于补充包被氨基酸组(P0.05); 补充包被EAA组对虾增重率显著高于补充包被EAA+NEAA组(P0.05)。饲料系数的变化正好与增重率变化相反(P0.05)。饲喂高蛋白质水平饲料较之饲喂低蛋白质饲料明显提高对虾增重率、虾体蛋白含量(P0.05), 但降低虾体脂肪含量(P0.05)。包被氨基酸组凡纳滨对虾血浆游离氨基酸含量总体显著低于CAA组(P0.05)。除谷氨酸、甘氨酸以及脯氨酸外, 各组对虾肌肉氨基酸含量无显著差异(P0.05)。结果表明, 在32%饲料蛋白质水平下, 用鸡肉粉完全替代鱼粉时, 饲料中补充包被EAA可明显促进凡纳滨对虾的生长, 且达到了鱼粉组的饲喂效果。     

The effect of organic diets on the performance of pullets maintained under semi-organic conditions

The effects of organic diets, with or without supplements of betaine, saponin, fructo-oligosaccharide and methionine, on the health, performance and gut flora of pullets were investigated. A comparison was also made between birds fed organic diets and those fed a non-organic diet. Day-old Lohmann Tradition pullets were reared in 24 groups of 64 chicks indoors until 11 weeks, and then in 48 groups of 24 to 27 chicks with access to range until 17 weeks of age. Groups of birds were fed one of eight diets, a conventional rearing diet with supplementary amino acids, an organic basal diet, organic basal plus methionine and organic basal supplemented with one of the test ingredients. At most stages of growth the birds fed the conventional diet and those fed the basal diet with methionine performed better than those that had no supplemental methionine. Other dietary treatments had no consistently significant effect on growth, the microbial populations within the gastro-intestinal tract of the birds or the number of parasite eggs excreted. After 5 weeks with access to range, the birds that were fed three out of five diets regarded as deficient in sulphur amino acids achieved similar weights (P > 0.05) to birds that received diets adequate in sulphur amino acids. Health and welfare of birds fed organic diets was not adversely affected; however, an investigation of birds housed in larger flocks and taken into the laying phase, when physical demands on birds are greatest, is required.     

Effects of Sulfur-containing Amino Acids and Glycine on Plasma Cholesterol Level in Rats Fed on a High Cholesterol Diet

The effects of the addition of individual amino acids on methionine-induced hypercholesterolemia (experiment 1), and the interacting effects of dietary protein level and sulfur-containing amino acids and glycine on plasma cholesterol concentration (experiment 2) were studied in growing rats fed on a high cholesterol diet. In experiment 1, rats were fed on a 25% casein-0.75% methionine (25CM) diet containing 2.5% of individual amino acids for 2 weeks. Methionine-induced hypercholesterolemia was prevented by the concurrent addition of glycine or serine, but the other amino acids tested (alanine, threonine, leucine, phenylalanine, lysine, arginine, and glutamic acid) had no effect. Histidine rather enhanced the hypercholesterolemia. In experiment 2, rats were fed on a 10%, 25%, or 50% casein diet containing 0.75% methionine, 0.60% cystine, 0.63% taurine, 2.5% glycine, or 0.75% methionine +2.5% glycine for 3 weeks. Dietary addition of 0.75% methionine increased the plasma cholesterol concentration for the 25% and 50% casein diets, but it decreased the plasma cholesterol for the 10% casein diet. When the addition level of methionine was doubled in the 10% casein diet, the plasma cholesterol concentration was significantly higher for the 1.5% methionine-added diet than for the 0.75% methionine-added diet. Cystine and taurine lowered plasma cholesterol for all dietary casein levels. Methionine-induced hypercholesterolemia with 25% and 50% casein diets was prevented by the glycine supplementation. These data suggest that sulfur-containing amino acids and glycine are important in plasma cholesterol regulation.     

Metabolic adjustments to varying protein intake in harbor seals (Phoca vitulina): evidence from serum free amino acids

Effects of varying dietary protein intake on serum free amino acid (FAA) concentrations were studied in harbor seals (Phoca vitulina) fed two different prey fish diets: either exclusively low-fat, high-protein walleye pollock (Theragra chalcogramma) or high-fat, relatively high-energy-density Pacific herring (Clupea pallasi). Significant differences in FAA concentrations and patterns were observed between the two diets. All essential amino acids (EAA), except methionine and phenylalanine, and two nonessential amino acids (NEAA), glycine and tyrosine, decreased when the diet was switched from herring to pollock and increased on switching back to herring. Both total EAA concentrations and EAA : NEAA ratios decreased with the elevated protein intake typical of a low-fat pollock diet, indicating an inverse correlation between EAA concentrations and dietary protein intake levels. We propose that differing dietary protein intake, caused by differences in macronutrient composition of the two prey fish species, induced a change in protein metabolism that was reflected in blood-circulating amino acids. These findings suggest that surveys of amino acid profiles may be useful to partially determine the protein metabolic status of harbor seals.     

Impaired translation initiation activation and reduced protein synthesis in weaned piglets fed a low-protein diet

Weanling mammals (including infants) often experience intestinal dysfunction when fed a high-protein diet. Recent work with the piglet (an animal model for studying human infant nutrition) shows that reducing protein intake can improve gut function during weaning but compromises the provision of essential amino acids (EAA) for muscle growth. The present study was conducted with weaned pigs to test the hypothesis that supplementing deficient EAA (Lys, Met, Thr, Trp, Leu, Ile and Val) to a low-protein diet may maintain the activation of translation initiation factors and adequate protein synthesis in tissues. Pigs were weaned at 21 days of age and fed diets containing 20.7, 16.7 or 12.7% crude protein (CP), with the low-CP diets supplemented with EAA to achieve the levels in the high-CP diet. On Day 14 of the trial, tissue protein synthesis was determined using the phenylalanine flooding dose method. Reducing dietary CP levels decreased protein synthesis in pancreas, liver, kidney and longissimus muscle. A low-CP diet reduced the phosphorylation of eukaryotic initiation factor (eIF) 4E-binding protein-1 (4E-BP1) in skeletal muscle and liver while increasing the formation of an inactive eIF4E.4E-BP1 complex in muscle. Dietary protein deficiency also decreased the phosphorylation of mammalian target of rapamycin (mTOR) and the formation of an active eIF4E.eIF4G complex in liver. These results demonstrate for the first time that chronic feeding of a low-CP diet suppresses protein synthesis in animals partly by inhibiting mTOR signaling. Additionally, our findings indicate that supplementing deficient EAA to low-protein diets is not highly effective in restoring protein synthesis or whole-body growth in piglets. We suggest that conditionally essential amino acids (e.g., glutamine and arginine) may be required to maintain the activation of translation initiation factors and optimal protein synthesis in neonates.     

High-Casein Diet Suppresses Guanidinoacetic Acid-Induced Hyperhomocysteinemia and Potentiates the Hypohomocysteinemic Effect of Serine in Rats

To determine the effect of dietary protein level on experimental hyperhomocysteinemia, rats were fed 10% casein (10C) and 40% casein (40C) diets with or without 0.5% guanidinoacetic acid (GAA) for 14 d. In addition, rats were fed 10C + 0.75% methionine (10CM) and 40C + 0.75% methionine (40CM) diets with or without 2.5% serine for 14 d to determine the relationship between the dietary protein level and intensity of the hypohomocysteinemic effect of serine. GAA supplementation markedly increased the plasma homocysteine concentration in rats fed with the 10C diet, whereas it did not increase the plasma homocysteine concentration in rats fed with the 40C diet. Although serine supplementation significantly suppressed the methionine-induced enhancement of plasma homocysteine concentration, the decreased plasma homocysteine concentration was significantly lower in rats fed with the 40CM diet than in rats fed with the 10CM diet. The hepatic cystathionine β-synthase and betaine-homocysteine S-methyltransferase activities were significantly higher in rats fed with the 40C or 40CM diet than in rats fed with the 10C or 10CM diet, irrespective of supplementation with GAA and serine. These results indicate that the high-casein diet was effective for both suppressing GAA-induced hyperhomocysteinemia and potentiating the hypohomocysteinemic effect of serine, probably through the enhanced activity of homocysteine-metabolizing enzymes.     

Post-ingestive food-aversion learning to amino acid deficient diets by the terrestrial slugLimax maximus

Summary An agar-based artificial diet containing carbohydrates, fats and twenty amino acids was constructed (Fig. 1). This diet is highly palatable and nutritionally complete forLimax maximus as demonstrated by significant ingestion on first encounter, consistent ingestion on subsequent days and good growth of young slugs fed this diet. Removing methionine, an essential amino acid, from the complete diet produces a food which is initially as palatable as the complete diet, but after one day's intake the amount of this deficient diet eaten is greatly reduced (Fig. 2). Removing alanine, a nonessential amino acid, does not produce any decrement in feeding relative to the complete diet (Fig. 5).A single meal can be sufficient for establishing the aversion to the deficient diet (Fig. 5). Following seven days of feeding on the deficient diet the aversion is retained with little or no attenuation for at least 30 days (Fig. 3) and does not generalize to either a known safe food (Fig. 3) or a novel food (Fig. 4). Evidence of a mild neophobia towards the artificial diet which attenuated after one or two meals was seen (Fig. 5).The learned aversion to the deficient diet is reversible if slugs are repeatedly fed the complete diet following feeding on the deficient diet. Also, slugs initially fed the complete diet will develop an aversion to the methionine-deficient diet after sampling it (Fig. 7).Slugs readily ate the artificial diets when these were offered 7 days post-hatch. The methionine-deficient diet however was not eaten in large amounts after the first meals and did not support growth (Fig. 9). Baby slugs fed the methionine-deficient diet for 10 days and then maintained on rat chow ate only small amounts when the deficient diet was presented again 126 days later, while baby slugs fed the complete diet or an alanine deficient diet for 10 days ate large amounts when these diets were presented 126 days later (Fig. 11 b).Supplementing the methionine-deficient diet with an injection of methionine into the haemocoel one hour after the completion of a meal completely blocks the development of a learned aversion while injection ofLimax saline does not (Fig. 8).These results are best explained by the hypothesis that the slugs acquire, post-ingestively, an aversion to the taste and probably the odor of the diet as the result of associative learning. The results of the experiments reported here indicate that there are substantial parallels at the behavioral level between mollusc and mammal with respect to post-ingestive feedback learning.     

Effects of protein level, methionine supplementation and carbohydrate type of the diet on liver lipid and plasma free threonine contents in the lactating rat

Eight groups of 13-15 female rats were fed purified diets after littering. Four groups received a low protein (8% casein) diet (groups 8) and the others, a normal protein (20% casein) diet (groups 20). Carbohydrates were supplied either as starch (groups S) or as starch plus 40% fructose (groups F). Half the animals received a 0.4% methionine supplementation (groups M). Four or five dams per group were sacrificed on days 2, 7 and 14 after littering. The diet intake was increased by methionine supplementation, substitution of starch for fructose and increased protein content, mainly during the second week of lactation. This influenced weight variation of the dams and litter growth. On all days, the plasma levels of cholesterol esters, triglycerides and phospholipids were positively correlated with the dietary protein level. On days 7 and 14, the liver neutral lipid content was increased in rats fed the low protein diets supplemented with methionine (groups 8SM and 8FM) and the normal protein diets containing 40% fructose (groups 20F and 20FM). The plasma free threonine content was positively correlated with the protein level in the diet. On day 14, rats fed a low protein diet had a threonine deficiency, except those in groups 8S and 8F. The plasma free threonine content of these rats was not reduced, possibly due to an impaired utilization of this amino acid. The liver lipidosis observed during lactation, in contrast to that observed during growth with a low protein diet, was not due to a threonine deficiency.     

Tryptophan Metabolism of Rats Fed on an Amino Acid Diet Devoid of One Branched Chain Amino Acid

In an attempt to study on metabolic changes in rats fed on an amino acid diet devoid of one branched chain amino acid and of niacin, rats were force-fed a leucine-free, isoleucine-free, valine-free or complete amino acid diet for 3 or 4 days and killed 3 hr after the feeding on day 4 or 5 to observe the body weight changes, the urinary nitrogen and N¹-methylnicotinamide (MNA), and liver tryptophanpyrrolase (TPase) and tyrosine-α-keto-glutarate transaminase (TKase) activities.

The excretion of the urinary nitrogen and MNA, TPase and TKase activities, and fat content of livers of rats force-fed these amino acid deficient diets were higher than those fed the complete amino acid diet. It was further confirmed in the present study that changes in TPase activity of rats given diets devoid of one essential amino acid were in the same direction with changes in urinary MNA which was observed in the previous studies on rats given threonine-free, tryptophan-free, methionine-free, lysine-free and complete amino acid diets. However, such metabolic changes in rats fed the leucine-free diet were not so remarkable, compared with those of rats fed the other amino acid deficient diets.     

Protection of adrenocortical activity by dietary casein in ether anaesthetized rats

Adrenal delta5-3beta-hydroxysteroid dehydrogenase (delta5-3beta-HSD) activity and serum corticosterone level were significantly higher in rats fed with 5% casein or 4% albumin diets after 1 hr of ether anaesthetic stress as compared to the controls, 5% casein and 20% casein (equivalent to 4% albumin) respectively. Ether anaesthesia to 20% casein fed rats caused no change in adrenal delta5-3beta-HSD activity and serum corticosterone level when compared with controls fed 20% casein diet. The results suggest that high milk protein diet may prevent acute stress effects by protecting adrenocortical activity. The present investigation opens up a new area of management of stress.