Effect of pre-feeding ammonium acetate on food intake of rats fed high protein diets

The effect on intake of a 75% casein diet after prefeeding for one week a 6% casein basal diet with additional 0%, 2%, 5%, 8% or 15% ammonium acetate was examined in rats trained to eat in three hours per day. Food intake was measured from 0-15, 15-30, 30-90, and 90-180 minutes for the first two days that the ammonium acetate diets were presented. Rats eating 5% and 8% or 15% ammonium acetate diet depressed their intake significantly for one day and for four days respectively. Rats eating 2%, 5%, 8%, or 15% ammonium acetate diets depressed their intake significantly from 0-30 minutes. When presented with the 75% casein diet, rats prefed 0% to 5% and 8% and 15% ammonium acetate diets ate 55% to 58% and 72% and 94% of their respective baseline intakes. It is suggested that prefeeding 15% ammonium acetate apparently induces sufficient metabolic adaptation to ammonia intake so that the rat is able to offset the metabolic consequences of intake of the 75% casein diet, thus preventing the usual food intake depressing effect of the high protein diet.     

Increase in plasma ammonia and amino acids when rats are fed a 44% casein diet

Rats were trained to eat a 6% casein basal diet during a 3-hour period per day. They were then fed either the same 6% casein diet or a 44% casein diet for 3 hours. No food intake depression was observed in the rats eating 44% casein diet during the 3-hour period. Plasma ammonia and amino acids and brain amino acids were measured at 0, 4, 12 and 24 hours after presentation of the 6% or 44% casein diets. Plasma ammonia rose to 134 (p less than 0.01) and 110 micromolar (p less than 0.05) in the 44% casein fed rats at 4 and 12 hours, respectively, as compared to 67 and 53 micromolar, respectively, for the 6% casein fed rats. All plasma amino acid concentrations except methionine and glutamate were elevated (p less than 0.05) at 4 hours. In the brain, threonine, glutamine and tyrosine concentrations were elevated (p less than 0.05) at 4 hours after diet presentation. At 24 hours, valine, isoleucine, leucine, phenylalanine, and methionine concentrations were also elevated (p less than 0.05). Because intake of the 44% casein diet decreases the second day of its presentation, as noted in an earlier experiment, the increases in plasma ammonia and its possible entry into the brain as reflected by increased brain glutamine together with changes in amino acid concentrations should be considered collectively among possible metabolic signals affecting intake of high protein diets.     

Protein selection by rats adapted to high or moderately low levels of dietary protein.

After preliminary studies on flavor acceptability, patterns and indices of subsequent feeding behavior were monitored by computer in young rats which were adapted to 15% or 70% casein diets before being offered, sequentially, choices between flavored diet pairs in which the proportions of percentage casein were 5/65, 5/55, 5/45, 5/35 and 5/25. Similarly adapted rats received these choices in the reverse sequence. Rats adapted to 15% casein usually ate randomly from the diet pairs and selected approximately 15-30% casein; individual behaviors were prominent. The 70% casein groups avoided the higher casein diet, often within minutes (except for the first-offered 5/25 choice), and seldom selected more than 10% casein; individual differences were infrequent. Such rats also distinguished between flavored 70% and 65% casein diets. Sizes and numbers of meals and rates of eating differed for the paired diets, especially for rats adapted to 70% casein. A flavor added to the 70% casein adaptation diet was not avoided when present only in the 5% casein diet of a 5/65 choice. Rats adapted to 70% soy protein before receiving flavored 5/65 to 5/25 choices selected 20-28% soy protein, a level far above those of casein selections by rats adapted to 70% casein. Dietary adaptation and type of protein thus affect subsequent diet selection and feeding patterns and indices.     

Olfactory self-selection of protein-containing foods

When one unflavored, nonprotein diet was available in two differently scented bins, rats fed a protein-free diet over four days ate more from the bin smelling of gluten, ovalbumin, yeast or fibrin, but not soy, casein or lactalbumin, than from the bin smelling of butter. Rats fed a protein-containing diet over the same four-day period had no such preference. This result demonstrates that protein-deprived rats can use odor cues in making their selection of certain proteins. Since the direction, speed, and size of preference for these protein odors, excepting soy, are remarkably similar to those previously observed when rats actually consumed the proteins, olfactory stimuli appear to elicit appropriate protein selection responses independently of other protein quality variables such as taste, texture or nutrient composition.     

Effects of monosodium glutamate on eating and drinking behavior in rats

The effect of monosodium glutamate (MSG) on eating and drinking behavior of rats was studied by means of preference tests in free choice situations. Low or high protein casein diets containing up to 3% MSG were selected indifferently compared to plain casein diet in 7 day trials. The acceptability of diets containing 7% MSG, particularly those of low protein level, was significantly reduced. Except for Day 1, the proportional intake (MSG flavored diet intake/total diet intake) of 9% protein--7% MSG diet was consistently lower than 18% protein--7% MSG diet. In brief exposure tests (10 min) solutions of MSG (0.02-8%) were preferred over deionized water. The acceptance of solutions containing higher concentrations of MSG was significantly reduced. The total volume intake of both choices (MSG flavored water and water) was significantly increased in tests using solutions containing 3-8% MSG. In long-term tests (1-14 days), solutions containing 0.05-1% MSG were preferred over water. The acceptance of a solution containing 5% MSG was significantly reduced. For maximum intake of moles of MSG, rats selected solutions which offered 3-5 x 10(-1) M MSG whether the experimental period was 10 min or up to 2 weeks. In a series of two choice preference tests using solutions where Na+ content and pH were kept equal, MSG was preferred over sodium acetate and over sodium glutamate but was less preferred than monosodium aspartate. The results of this study can be explained in terms of sensory quality.     

Responses to protein dilution in the adult female rat

Adult female rats were given ad lib access to 3 food cups containing casein, cornstarch, and a fat mixture, or a composite diet formulated from these nutrients. One-half of the animals in each diet condition were subjected to 25, 50, 150, and 500% sequential dilutions of the casein or the composite diet. Self-selecting rats decreased intake of the diluted casein, but increased consumption of cornstarch, so that total caloric intake remained constant. Composite-fed rats compensated for dilution by increasing the volume of diet consumed, but caloric regulation was less precise than in self-selectors.     

Dietary soy protein and isoflavones: minimal beneficial effects on bone and no effect on the reproductive tract of sexually mature ovariectomized Sprague-Dawley rats

OBJECTIVE: The present study was conducted to determine the effects of dietary soy protein and isoflavones on bone and the reproductive tract in the absence of the ovary. DESIGN: Three-month-old Sprague-Dawley rats (n = 56) were either sham-operated or ovariectomized and then fed diets containing casein or soy protein +/- isoflavone extract for 12 weeks. The amounts of casein, soy protein, and extract (per kg diet) in each group were as follows: (1) Ovariectomy, 200 g of casein; (2) Ovariectomy+low soy, 100 g of casein + 100 g of soy protein; (3) Ovariectomy+high soy, 200 g of soy protein; (4) Ovariectomy+low extract, 200 g of casein + 17.2 g of extract; (5) Ovariectomy+high extract, 200 g of casein + 34.4 g of extract; (6) Ovary intact, 200 g of casein; (7) Ovariectomy+estradiol-17beta, 200 g of casein. Diet consumption, body weight, uterine weight, urine deoxypyridinoline, and bone mineral density of the femur and lumbar vertebrae were measured. The femur rigidity was evaluated by histomorphometry. The reproductive tract (uterus, vagina, and cervix) was studied histologically. RESULTS: The Ovariectomy group showed significant increases in body weight, diet consumption, and deoxypyridinoline, decreases in uterine weight and bone mineral density, and negative changes in histomorphometry compared with the Ovary intact group. Neither soy protein nor extract diets abrogated these alterations, except for the Ovariectomy+high extract group that showed statistically significant positive changes in histomorphometric parameters. There were no histological differences in the reproductive tract among Ovariectomy, Ovariectomy+soy, and Ovariectomy+extract groups. The estradiol-17beta replacement abrogated ovariectomy-induced alterations. CONCLUSION: Dietary intake of isoflavones by sexually mature ovariectomized rats has a minimal beneficial effect on bone with no effect on the reproductive tract.     

Protein selection, food intake, and body composition in response to the amount of dietary protein

Though not universally observed, moderately low-protein diets have been found to increase caloric intake and body fat. It appears that animals overeat in calories in order to obtain more dietary protein. For animals to control protein intake, they must be able to distinguish between two isocaloric diets containing different percentages of protein and make the appropriate dietary selection on the basis of their previous history of protein intake. Experiment 1 examined the 24-h diet selection (5 vs. 35% casein) of Sprague-Dawley rats that had been previously fed diets containing various percentages of dietary protein (5, 10, 20, 35, or 60% casein). Animals fed 5, 10, or 20% dietary protein showed a preference for the higher protein selection diet. In contrast, no significant diet preference was found in animals pre-fed the two higher levels of dietary protein (35 or 60% casein). In this study, daily food intake and body fat of rats fed the low-protein diets (5 and 10% casein) were similar to rats fed the 20% casein diet. Experiment 2 examined the effects of the level of methionine supplementation on rats fed 10% casein. In this study, food intake and body fat were increased by approximately 20% in rats fed 10% casein diets, regardless of the level of methionine supplementation (0.3 vs. 0.15%). Together, the results suggest that the presence of low-protein-induced hyperphagia helps maintain body protein levels in the face of moderately low dietary protein and promotes an increase in the amount of body fat and energy.     

Dietary soy protein and isoflavones have no significant effect on bone and a potentially negative effect on the uterus of sexually mature intact Sprague-Dawley female rats

OBJECTIVE: To evaluate the effect of dietary soy protein and isoflavones on bone and the reproductive tract in premenopausal rats. DESIGN: Three-month-old intact Sprague-Dawley female rats (N = 50) were fed diets containing casein, soy protein, or casein with isoflavone extract for 12 weeks. The amount of casein, soy protein, and extract (per kilogram diet) in each group was: (1) 200 g casein (control); (2) 100 g casein plus 100 g soy protein (low soy); (3) 200 g soy protein (high soy); 4) 200 g casein plus 17.2g extract (low extract); and (5) 200 g casein plus 34.4 g extract (high extract). Diet consumption, body weight, uterine wet weight, urinary deoxypyridinoline concentration, and bone mineral density of the femur and lumbar vertebrae were measured. Femur rigidity was evaluated by histomorphometry. The uterus and vagina were studied histologically. RESULTS: Rats in all treatment groups had lower body weights and lower deoxypyridinoline concentrations compared with controls, but none of the differences was statistically significant. There was no significant difference in femur and lumbar bone mineral density, uterine wet weights, or histomorphometry between the control and treatment groups. Histologically, uteri and vaginae were normal in all groups except that 1 of 10 rats in the high-soy group and 2 of 10 rats in the high-extract group showed extensive squamous metaplasia in the uterine gland. CONCLUSION: These results suggest that dietary isolated soy protein and isoflavones have no effect on bone and the vagina during premenopausal period, but may have an adverse effect on the uterus.     

Effects of dietary protein on food and water intake in spontaneously hypertensive rats

We have been studying the development of hypertension in spontaneously hypertensive rats (SHR) fed a low protein diet. The effects of a low protein diet upon food and water intake were examined. Body weight gain, food and water intake were measured in three to twenty-three week-old SHR and Wistar Kyoto rats (WKY) fed diets containing 8%, 15% or 25% casein. Body weights of SHR and WKY fed an 8% casein diet were significantly lower at 23 weeks than rats on the higher protein diets, although both groups on the 8% diet consumed more food and water per g of body weight. In addition, SHR fed an 8% casein diet drank less water per gram of food than WKY or SHR fed 15% and 25% casein diets. These results indicate that changes in food and water intake, as a consequence of low protein diets, should be an additional consideration when examining the effects of dietary protein on the development of hypertension.     

Effect of melatonin on total food intake and macronutrient choice in rats

Melatonin, a hormone secreted in a rhythmic manner over 24 h mainly by the pineal gland, is used to alleviate the symptoms of jetlag and treat sleeping problems. The objective of the present study was to examine the effects of a 7-h phase-shift from the natural peak of melatonin secretion on total food intake and macronutrient selection. Forty-eight adult Wistar rats of both sexes were divided in three dietary groups, each group offered a simultaneous and different choice of a carbohydrate- and a protein-rich diet. Macronutrient intakes following intraperitoneal administration of four doses of melatonin (3000, 6000, 10 000 and 15 000 pg/ml blood) at dark onset were examined. Melatonin increased short- (4 h postinjection) and long-term (12 h postinjection) nocturnal total food intake in both male and female rats, mainly with the two highest doses. This effect of melatonin was mainly due to a short-term increase of intake across all carbohydrate-rich diet preparations (dextrin/cornstarch, cornstarch, and sucrose/cornstarch) and across genders. This consistent effect of melatonin on the intake of carbohydrate-rich diets with contrasting sensory attributes rules out the possibility that melatonin acts on sensorymotor pathways, thus suggesting that melatonin's effect on food intake is controlled by the carbohydrate content of the diet. In contrast, melatonin could be affecting some sensory or motor processes peculiar to the ingestion of protein since it increased protein-rich diet intake inconsistently across the various preparations (casein, soy isolate, and egg protein) as well as genders. This evidence supports the view that melatonin acts as a time indicator, reinforcing the animals with a “night cue”, and favors predominant carbohydrate intake normally occurring at the beginning of the activity period.     

Maternal malnutrition in the rat: Effects on food intake and body weight

The effects of dietary protein level on food intake and body weight were examined in adult female rats during a 35-day pre-mating period and during gestation and lactation. During the pre-mating period, no differences in daily food intake were observed among rats fed a 6% casein, 8% casein or 25% casein diet. However, during this period, rats fed the 6% casein diet gained significantly less weight than those with ad lib access to the 8% or 25% casein diets or than rats pair-fed the 25% casein diet in amounts equivalent to that consumed by rats in the 6% or 8% casein groups. Additionally, rats fed the 6% casein diet displayed decreased efficiency of energy utilization, calculated as weight gain per 100 kilocalories consumed, relative to rats fed the 8% or 25% casein diets. No differences in food intake were observed among the groups during gestation. However, rats fed the 6% casein diet gained less weight than rats fed the 8% or 25% casein diets. During lactation rats fed either the 6% or 8% casein diet consumed significantly less food than animals given the 25% casein diet ad lib. During the second week of lactation, rats receiving ad lib access to the 25% casein diet gained weight while those receiving the 6% or 8% casein diets continued to lose weight. At parturition, body weights of pups did not differ as a function of dietary condition. However, by day 12 of life, pups whose dams had ad lib access to the 25% casein diet weighed significantly more than pups whose dams consumed the 6% or 8% casein diet or whose dams were pair-fed the 25% casein diet in amounts equivalent to those consumed by rats fed the 6% or 8% casein diet.     

Relationship of dietary fat content to food preferences in young rats

Weanling rats were fed either a high-fat (30% of calories) or a low-fat (10% of calories) diet for four weeks, after which fat preference was assessed using a choice paradigm. Fat preference was measured during 2-hour intake tests in which three peanut butter/peanut oil mixtures containing 0.50, 0.61, and 0.71 grams fat/gram were offered to each animal. Rats fed the high-fat (HF) diet preferred the highest-fat mixture and consumed more total fat during intake tests than animals fed the low-fat (LF) diet. Intake of NaCl and sucrose solutions was measured during separate intake tests. LF-fed rats drank more NaCl solution than HF-fed rats. Following these tests a subgroup of the LF-fed animals was fed the HF diet, and a subgroup of the HF-fed group was fed the LF diet for a further four weeks. Upon repetition of the intake tests, rats that had been fed the HF diet during the initial four weeks still preferred the highest-fat mixture.     

The effects of early dietary experience on subsequent protein selection in the rat

Two series of experiments were designed to investigate the effects of protein composition of the first solid food consumed by young rat pups on their subsequent protein intake after weaning. In the first series of experiments, pups were allowed access to the maternal diet between 17 and 21 days of age, then weaned to a choice of 10% and 60% casein diets at 21 days. Pups with access to a maternal diet containing 39% casein selected high levels of protein (calculated as % of total calories selected as protein: 50 +/- 1% (access) vs. 29 +/- 3.5% (no access), p less than 0.05), whereas lower levels of protein were selected following access to a maternal diet containing 26% casein (16 +/- 1.1% (access) vs. 23 +/- 1% (no access), p less than 0.05) or 10% casein (27 +/- 3.9% (access) vs. 39 +/- 3.7% (no access), p less than 0.05). In a second series of experiments, the influence of early diet composition on subsequent protein selection was confirmed by prematurely weaning rats at 17 days of age to either 39% or a 10% casein diet. When given a choice of 10% and 60% casein diets at 21 days, pups selected higher levels of protein if weaned to a 39% casein diet as compared to a 10% casein diet (37 +/- 5.0% vs. 19 +/- 3.5% (p less than 0.05) for the first experiment and 43 +/- 3.0% vs. 26 +/- 4.0% (p less than 0.05) in a second experiment). It was concluded that the level of protein selected by young rats is positively correlated with the protein concentration of their first solid food.     

Self-selection of dietary casein and soy-protein by the cat

Growing specific-pathogen-free kittens were fed for two weeks a choice between two complete diets differing only in protein content. When casein diets containing 18, 36 and 54% protein were offered in the three possible combinations, the kittens consistently avoided the higher casein diets and kittens offered the two highest levels of casein significantly reduced their total food intake. In one soy-protein choice study, 16, 31 and 63% protein diets were each offered with a protein-free (PF) diet. When diets were similar in physical consistency, kittens selected similar amounts of both diets with the result that the PF:16% group consumed below their requirement of protein. In another soy-protein experiment the 16, 31 and 63% protein diets were offered in their three possible combinations. Kittens in all three groups selected similar amounts of both diets. Except for their avoidance of casein, the kittens did not regulate in a consistent manner their intake of protein and therefore, behaved very differently from the rat in the self-selection of dietary protein.     

Unlearned specific appetite for protein

Appetite for most protein appears to be unlearned. Protein-depleted rats, in the first half hour of a test, chose a significantly larger proportion of soybean, gluten, zein, fibrin, yeast torula and ovalbumin than protein replete controls, but not of casein or lactalbumin. There had been no opportunity to learn such dietary preference before the test. The preference for the protein in protein-depleted rats manifested itself significantly from the first minute, and also in females in which protein need was induced by pregnancy rather than a diet different from the controls.     

Hepatic vagotomy disrupts somatotropin-induced protein selection

Rats treated with somatotropin (STH) and allowed to self-select between diets varying in protein content will consume more of the high-protein diet. The objective of this study was to determine the role of the hepatic vagus nerve in this ability to select protein. Female Sprague-Dawley rats (n=40) received a hepatic vagotomy (HVAGX) or a sham surgery. Postsurgery, the rats were maintained on pelleted diets for 2 weeks, after which the rats were adapted to selecting between powdered diets with 5% casein and 30% casein. After a 7-day adaptation to diet selection, rats in each surgical treatment group were treated with STH (4 mg/day) or physiological saline for 14 days. Body weight and intake were recorded daily. STH treatment increased growth rate to a similar degree in both sham and HVAGX groups. Despite causing an increase in total food intake, there was no effect of HVAGX alone on body weight. Relative to the sham-saline group, sham-STH in treated rats had greater total food intake that was accounted for entirely by increased consumption of the 30% protein diet and no change in intake of the 5% diet. In contrast, HVAGX+STH rats exhibited 20-30% increases in consumption of both the 5% and 30% protein diets. Thus, the HVAGX+STH rats recognized an increased need for protein, but were unable to distinguish between the high- and low-protein diets and selected more of both. The data suggest that the ability to alter diet selection in response to a stimulation of protein accretion is at least partially mediated through the liver and hepatic branch of the vagus nerve.     

Effects of a soy protein product on serum and tissue cholesterol concentrations in swine fed high-fat, high-cholesterol diets

Hypocholesterolemic effect of a soy protein product was studied in swine fed a high-fat, high-cholesterol diet. In the first experiment, a group of swine were fed 42% butter (by calories) and 1055 mg cholesterol daily, with casein as the source for protein, for 6 weeks and this diet resulted in moderately high serum cholesterol concentrations (219 ± 33 mg/dl). Another group fed the same diet except with soy protein product as the protein source instead of casein showed virtual normocholesterolemia at the end (107 ± 3 mg/dl). Cholesterol balance was studied under non-steady state conditions using methods designed for this purpose. Reflecting the serum cholesterol concentration, the total body cholesterol concentration (excluding CNS) was also significantly lower in soy protein group. However, parameters of cholesterol balance, such as fecal neutral and acidic steroid excretions, dietary cholesterol absorption, and whole body cholesterol synthesis were studied and no differences were demonstrated between the casein- and soy protein-fed swine. The experiment was repeated and in Experiment II virtually the same results were obtained. When swine were given the same high-fat, high-cholesterol diets with $\text{1}\text{2}$ casein + $\text{1}\text{2}$ soy protein or casein + soy protein, hypocholesterolemic effects were also observed. Therefore, such action is probably caused principally by soy protein per se rather than simply by replacement of casein by soy protein. Addition of dl-methionine to soy protein containing diet did not alter the hypocholesterolemic effect of soy protein indicating that the effect was not the result of methionine deficiency. In conclusion, we can state that the hypocholesterolemic action of soy protein was clearly demonstrated in swine fed a high-fat, high-cholesterol diet, but that the mechanism of action is yet to be established.     

Effects of hippocampal lesions on adaptive intake of diets with disproportionate amounts of amino acids

Bilateral double electrolytic overlapping lesions were placed in dorsal-lateral hippocampus of male 230 g rats, and their food intake responses to the ingestion of diets containing disproportionate amounts of amino acids were examined. Rats with such lesions and intact control rats maintained their normal intakes of the 6% casein basal diet or a threonine basal amino acid diet postoperatively. However, they exhibited marked initial food intake depression, similar to that of intact rats, when fed the threonine imbalanced amino acid diet. Also, animals with lesions in certain areas of the dorsal-lateral hippocampus showed facilitated adaptation to the amino acid imbalanced diet. Similar severe reduction in food intake with relative lack of adaptation were observed in both the intact controls and rats with hippocampal lesions when fed amino acid diets completely devoid of threonine. Initial food intake of rats with hippocampal lesions was inhibited drastically as was the case with the intact controls when fed a 75% casein high protein diet. All rats, either intact or lesioned, showed similar slow adaptation patterns with the prolonged ingestion of the high protein diet. The initial food intake responses and facilitated adaptation of the animals bearing lesions in certain areas of the hippocampus suggest that such areas are not crucially involved in the inhibition of food intake of rats fed disproportionate amounts of dietary amino acids. Rather, such areas of lesions in the hippocampus may play a role in a system governing the behavioral adaptation of the intake of amino acid imbalanced diets but not of diets containing amino acids in general excess. This would also indicate that different mechanisms control the intake of amino acid imbalanced diets and diets containing amino acids in excess.