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 self-selection vs. complete diet: body weight gain and meal pattern in rats.

Food intake and body weight gain of male adult Wistar rats were examined in two groups of animals. One group (n = 14) was allowed to select its diet from separate sources of protein (casein, 3.1 kcal/g), fat (lard and sunflower oil, 7.9 kcal/g) and carbohydrate (CHO, starch and sucrose, 3.3 kcal/g). Another group (n = 10) received a nutritionally complete diet (3.3 kcal/g). After 2 weeks of adaptation to the diets, body weights and meal patterns were recorded for at least 4 days. The total caloric intake was nearly identical for the two groups of rats. Rats given dietary choice gained less weight over 4 days than rats fed chow and showed reduced feed efficiency. During the 24-h period, self-selecting rats consumed 20.8% of calories as proteins, 21% as fats and 58.2% as CHO. Self-selecting rats ate significantly less calories during the day than did rats given chow. The chow diet consisting of 17.3% calories as protein, 7.6% as fat and 75.1% as CHO. When comparing the self-selecting group nutrient intakes to those of chow-fed group it was observed that 24-h protein calorie intakes were identical in both groups. Fat intake was significantly higher and CHO reduced as compared to chow-fed rats. During the day, CHO intake was higher in self-selecting rats, and fat intake was not significantly reduced. During the night, protein and fat intakes were significantly higher in self-selecting rats, while CHO intake was significantly decreased, particularly in the last periods of the night.(ABSTRACT TRUNCATED AT 250 WORDS)     

Dietary fat-induced hyperphagia in rats as a function of fat type and physical form

The influence of dietary fat on food intake and weight gain was assessed by feeding adult female rats diets that differed in the type and form of fat, as well as in the availability of other macro- and micronutrients. Compared to chow-fed controls, the various fat diets increased total food intake by 4% to 27%. Specifically, rats fed chow and a separate source of fat (fat option diet) consumed more fat and total calories, and gained more weight when the fat source was emulsified corn oil rather than pure corn oil or was vegetable shortening rather than corn oil. However, corn oil and shortening had similar effects on caloric intake and weight gain when presented as emulsified gels. Also, pure and emulsified-gel forms of shortening did not differ in their effects on caloric intake and weight gain. Supplementing the vegetable shortening with micronutrients, however, enhanced its hyperphagia-promoting effect. The results of two-choice tests revealed that the rats' preferences for the orosensory properties of the various fat sources did not account for the differential hyperphagias obtained. Rather, it appears that long-term fat selection and caloric intake are influenced primarily by postingestive factors. Fat selection and total intake were determined not only by the fat source itself, but also by the other diet options. That is, rats selected more fat and consumed more calories when chow was the alternative food than when separate sources of carbohydrate and protein were available.     

Changes in macronutrient selection as a function of dietary tryptophan

It has been hypothesized that the serotonergic system is involved in the regulation of carbohydrate and/or protein intake. Tests of this hypothesis using added dietary tryptophan and diets varying in the ratio of carbohydrate/protein resulted in depressed intakes of high carbohydrate/low protein diets, elevated intakes of low carbohydrate/high protein diets, and a reduction of total caloric intake. The present studies gave rats increased options for adjusting to added tryptophan by providing them with separate sources of protein, carbohydrate, and fat. The results showed the expected decrease in carbohydrate intake, but also increases in fat intake and, to a lesser extent, protein intake. Total caloric intake was conserved. Hypothalamic concentrations of serotonin and 5-hydroxyindole acetic acid indicated increased activity of the serotonergic system. These results lend support to serotonin's involvement in nutrient selection, in that carbohydrate consumption decreased in response to tryptophan loading, but indicate that other nutrients may also be affected. Given the option of altering fat intake, the animals maintained a constant caloric intake despite the reduction of carbohydrate consumption.     

Developmental aspects of sucrose-induced obesity in rats

Daily caloric intakes and body weights were measured from weaning to 70 days of age in male Sprague-Dawley rats given access to either a standard laboratory diet and water, or the standard diet, a 32% sucrose solution and water. Lee index of obesity (3 square root body weight/naso-anal length) and fasting blood glucose levels were determined at 46, 57, and 70 days of age. Animals were sacrificed at 70 days, and body composition analyses were performed. Aniamls given access to the sucrose solution consumed significantly more calories per day than animals given only the standard diet. Sucrose animals took approximately 50 to 60% of their daily caloric intake from the sugar solution. Despite the greater caloric intakes of the sucrose animals, sucrose and control animals did not differ in body weight. While there were no differences in body weights between the two groups, the Lee Index of obesity was significantly greater in the sucrose animals than in controls as early as 46 days of age. Fasting blood glucose levels were significantly lower in sucrose animals than in controls at both 46 and 57 days of age. Direct determinations of body compositions when animals were 70 days of age revealed that animals with access to sucrose had significantly greater percentages of body fat and lower percentages of body protein than controls.     

Flavor-cued modulation of intake in rats: role of familiarity and impact on 24-h intake.

Following training with distinctively flavored nutritive solutions that differ in concentration and thus in caloric value, rats demonstrate flavor-postingestive consequence learning by preferentially consuming one of the flavors in two-bottle tests (both flavors in nutrient-identical solutions.) Experiment 1 investigated whether the relative familiarity of the flavor-nutrient combinations encountered in two-bottle tests contributes to the observed preference. One of the training concentrations (rather than the customary intermediate concentration) was used to present the flavors in testing; thus, one of the flavors was in a familiar context while the other was in an unfamiliar context. The results of two independent trials (rats trained with 1 and 5% sucrose; rats trained with 5 and 40% sucrose) confirmed that two-bottle test preference was not a preference for the familiar flavor-nutrient combination. Experiment 2 examined whether caloric expectancies based upon a previously learned flavor-postingestive consequence association would affect total daily intake. On alternating days, rats consumed 30 mL of dilute (5%) and concentrated (40%) sucrose, each distinctively flavored. When given 30 mL of 22.5% sucrose containing each of the flavors on separate test days, they ate less chow and thus fewer total calories over 24 h when given the flavor previously paired with concentrated sucrose. Experiment 3 replicated the design of Experiment 2 except that fat calories were used instead of sucrose; no significant adjustment of chow intake in extinction tests was noted, even when the number of fat calories used in training was increased (Experiment 4). Thus, rats did not exhibit flavor-cued modulation of chow intake when trained with fat, in contrast to responsivity to flavor cues when trained with sucrose. This differential responding to fat versus carbohydrate calories is consistent with previous observations, in a variety of paradigms, that modulation of caloric intake is less energetically appropriate when ingested foods are high in fat relative to high-carbohydrate foods.     

Modulation of oxidative DNA damage levels by dietary fat and calories

Decreased dietary intake of fat and/or calories generally results in a lower incidence of mammary gland tumors in rodents. Feeding of either low-fat or calorie-restricted diets to rats also has been shown to result in decreased levels of oxidative DNA damage. Since oxidative DNA damage is suggested to have a role in carcinogenesis, this may be one mechanism by which dietary change can reduce cancer risk. The effects of calorie-restricted diets on both oxidative DNA damage levels and mammary gland tumor incidence are generally more pronounced than that of low-fat diets. There is, however, some difficulty in defining what amount of fat should be used to prepare ‘low-fat’ and ‘high-fat’ rodent diets as well as what a suitable fat intake for control diets should be in studies that examine the effects of dietary fat and/or calories on tumorigenesis. In particular, the promoting effects of dietary fat may be exerted only up to a certain level of fat, above which no further effect is observed. Another difficulty in the interpretation of the results is that there may be a time-dependent effect of high fat diets on oxidative damage, with increased damage resulting only when the diets are fed for longer periods of time. The appropriate experimental approach to model human dietary exposures therefore remains to be determined. Although the effects of caloric intake on mammary gland tumorigenesis appear to be more pronounced than that of fat intake, low-fat diets still may be useful as a preventive measure in human populations to reduce breast cancer risk for individuals who cannot safely reduce their caloric intake.     

Patterns of nutrient selection in rats with streptozotocin-induced diabetes

The effects of experimental diabetes on energy intake, patterns of nutrient selection, water intake, body weight and body composition were examined in male Sprague-Dawley rats given ground Purina Chow or a dietary self-selection regime. Following adaptation to dietary conditions, a portion of the animals in each diet group were made diabetic by the administration of 45 mg/kg streptozotocin (STZ). The remaining animals in each group served as vehicle-injected controls. STZ reliably produced diabetes in rats on both dietary regimes. Immediately after the induction of diabetes, rats on the self-selection regime increased carbohydrate and protein intakes and decreased fat intake. Approximately three weeks after STZ administration, diabetic rats reduced carbohydrate intake and increased fat intake. Diabetic animals in both diet groups were hyperphagic and polydipsic relative to non-diabetic controls. During the first three weeks following STZ injections, energy and water intakes of diabetic animals in the two dietary conditions were similar. However, after this initial period, energy and water intakes of diabetic rats given the self-selection regime were significantly lower than those of diabetic animals given Purina Chow.     

Quantifying food intake in socially housed monkeys: social status effects on caloric consumption

Obesity results from a number of factors including socio-environmental influences and rodent models show that several different stressors increase the preference for calorically dense foods leading to an obese phenotype. We present here a non-human primate model using socially housed adult female macaques living in long-term stable groups given access to diets of different caloric density. Consumption of a low fat (LFD; 15% of calories from fat) and a high fat diet (HFD; 45% of calories from fat) was quantified by means of a custom-built, automated feeder that dispensed a pellet of food when activated by a radiofrequency chip implanted subcutaneously in the animal's wrist. Socially subordinate females showed indices of chronic psychological stress having reduced glucocorticoid negative feedback and higher frequencies of anxiety-like behavior. Twenty-four hour intakes of both the LFD and HFD were significantly greater in subordinates than dominates, an effect that persisted whether standard monkey chow (13% of calories from fat) was present or absent. Furthermore, although dominants restricted their food intake to daylight, subordinates continued to feed at night. Total caloric intake was significantly correlated with body weight change. Collectively, these results show that food intake can be reliably quantified in non-human primates living in complex social environments and suggest that socially subordinate females consume more calories, suggesting this ethologically relevant model may help understand how psychosocial stress changes food preferences and consumption leading to obesity.     

Food intake in rats is unaffected by the profile of dietary essential fatty acids

Food intake may be differentially responsive to the type of fat in the diet. The present investigation sought to evaluate the energy intake of rats maintained on either a low-fat or a high-fat diet mixed with an oil rich in either linoleic (18:2; n-6; safflower oil) or linolenic (18:3; n-3; flaxseed oil) acid. In Experiment 1, rats (n=28) consumed low-fat versions of either the safflower oil diet or the flaxseed oil diet, each at 9.28% fat (wt/wt). In Experiment 2, different rats (n=28) consumed high-fat versions of these diets, each at 23.6% fat (wt/wt). Within each experiment, the energy intake of rats receiving the safflower oil diet was compared to the energy intake of rats receiving the flaxseed oil diet. Food intake was measured under short-term, long-term and food-deprived conditions. In Experiment 1, short-term energy intakes were not different between the groups, thus demonstrating equal acceptance of the test diets. There were no consistent differences in long-term energy intakes between the safflower group and the flaxseed group. In addition, there were no differences in energy intake under the food-deprivation condition. Results from Experiment 2 paralleled those of Experiment 1. Taken together, the present results suggest that the essential fatty acid profile of the maintenance diet does not influence food intake when nutritive oils are the predominant fatty acid source.     

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.     

Dilutions of low- vs. high-fat diets on intakes and gastric volumes in rats

Rats adapt to changes in dietary energy to maintain nearly constant energy intakes. This regulation indicates that animals sense and respond to nutrient content. We sought to determine whether this response was affected by the fat content of the diet. Our second goal was to determine how energy dilution affected intragastric volumes. Rats were randomized to high (18% w/w) and low fat (4.5% w/w) as the energy density of the diet was altered from 2.0 to 3.5 kcal/g. Average energy intake during 7-h feeds rose steeply (P<.01) when density was increased from 2.0 to 3.0 kcal/g, but modestly as density increased from 3.0 to 3.5 kcal/g. In other rats on 18% vs. 32% fat diets, energy intakes increased significantly (P<.01) as density of the diet was raised from 3.5 to 4.5 kcal/g. During diets at 2.0 and 2.5 kcal/g, animals on 18% fat ate fewer kilocalories than those on 4.5% fat; but over 3.0-4.5 kcal/g, energy intake was similar regardless of fat concentration (4.5-32%). Gastric contents after 7-h feeds increased with grams of food ingested similarly for high- and low-fat diets. We concluded that in rats: (a) compensation to energy dilution or concentration was inexact but (b) was about equal for high- vs. low-fat diets; thus, high fat was as well sensed as high carbohydrate; (c) compensations for energy densities were made despite varied gastric volumes; thus, rats learned to override the stimulus of gastric stretch and to sense energy via extra gastric mechanisms.     

Influence of early dietary experience on energy regulation in rats

The environmental mechanisms that have contributed to the rapid increases in overweight and obesity in the US over the past 25-30 years have yet to be fully specified. One hypothesis that has been forwarded is that increased consumption of calories in liquid form may be a contributing factor, since some studies support the idea that caloric compensation is less adequate for liquid calories compared to calories delivered in more solid form. Work from our laboratory using rats has examined the role that differences in diet viscosity may play in altering energy intake and body weight regulation. This work has suggested that when offered diets matched on caloric density, macronutrient and micronutrient composition, and differing only in viscosity, adult rats fail to compensate for calories delivered in low-viscosity form in short-term intake tests. Further, long-term consumption of low-viscosity diets leads to enhanced weight gain in adult rats. In the present studies, we examined whether short- or long-term exposure to varying relationships between viscosity and calories led to altered food intake or body weight regulation in juvenile rats. The results demonstrated that animals given either short- or long-term experience with direct relationships between viscosity and calories (high viscosity, high calorie meals and low viscosity, low calorie meals) did not differ in food intake or body weight gain compared to animals given short- or long-term experience with indirect relationships between viscosity and calories (high viscosity, low calorie meals and low viscosity, high calorie meals). When juvenile rats were given long-term (9 weeks) exposure to a single, high or low viscosity diet supplement, matched on caloric density and differing only in viscosity, there were no effects on body weight gain. However, analysis of body composition using DEXA demonstrated that animals consuming the low viscosity supplements had significantly greater body fat than animals that consumed either the high viscosity supplement or no dietary supplement at all. These differences in body fat persisted for at least 3 months following the cessation of dietary supplements; during this 3-month period, animals previously exposed to the low viscosity supplement also gained significantly more weight than animals previously exposed to the high viscosity supplement. Taken together, the results suggest that consuming calories in low viscosity form may contribute to poor intake regulation over the short-term and to increased adiposity over the long term. When animals experience these diets as juveniles, these effects may persist into adulthood.     

Caloric compensation in hypothalamic obese rats

The present study explored the influence of pelleted diets on adjustment to caloric dilution in hypothalamic obese rats. Medial hypothalamic lesioned and normal rats were maintained on a high fat diet until the static stage of hyperphagia was reached. They were given three pelleted diets which consisted of undiluted Noyes pellets and Noyes pellets diluted 25 or 50 percent with kaolin. In contrast to previous reports that hypothalamic obese rats do not compensate for caloric dilution of their diet, the obese animals showed as large an increase in food intake on the dilute pelleted diets as did the control animals. These results were interpreted by viewing texture as a dimension of palatability which influences food intake in hypothalamic obese rats.     

The preference-aversion behavior of rats for nutritionally-controlled diets containing oil or fat

The preference-aversion behavior of Sprague-Dawley rats for semi-purified diets containing various levels of either corn oil, animal fat (lard) or hydrogenated vegetable oil (Crisco) was determined in brief and long-term, two-choice preference tests. After the first day of exposure, rats selected more calories from the control-unadulterated diet than from experimental diets containing either oil or fat. Only in the first hour of exposure (with diets containing either 5% Crisco, 15 or 25% lard) or in the first day (with diets containing 5% Crisco, 5 or 15% lard) was a preference for experimental diets observed. There were significant effects of oil and fat concentration and of exposure time upon preference for experimental diets, such that preference for the experimental diets was reduced as levels of oil or fat and as the time of experiment increased. Changes in diet selection had no major effect on total caloric intake measured as the sum of both experimental and control dietary choices, although total intake on a daily basis tended to decrease during the middle portion (2-4 days) of the study, then recover by the end of the study. The results suggest that postingestional factors related to the dietary oil and fat (rather than the sensory properties of the diets) directed the long-term preference behavior of rats for semi-purified diets adulterated with various levels of either corn oil, animal fat (lard) or hydrogenated vegetable oil (Crisco).     

Diet composition determines course of hyperphagia in developing Zucker obese rats

Previous observations from this laboratory indicate that, during growth, the hyperphagia of the male genetically obese Zucker rat reaches a peak or "breakpoint" and then declines. To examine the effect of dietary macronutrient content on the course of hyperphagia, groups of male lean and obese rats were maintained from 5-28 weeks of age on powdered chow, or isocaloric diets (3.6 kcal/g) containing 72% of calories as corn oil, dextrose, or soy isolate protein (n = 5 lean and obese rats/diet). On chow, hyperphagia was maintained at a level of 7-8 g above lean control intake until a "breakpoint" was reached at 17 weeks, and obese intake declined to lean control level. On the fat diet, hyperphagia was increased to 10 g/day when a breakpoint was reached at 8 weeks. On the dextrose and protein diets, hyperphagia at a level of 3-4 g/day reached breakpoints at weeks 18 and 16, respectively. On all diets, the intakes of obese rats were precisely equal to the intakes of lean control rats by weeks 19-20. These data show that the magnitude and duration of hyperphagia in the developing obese rat are influenced by diet composition. Previously, we have proposed that the obese rat's hyperphagia arises from rapid adipocyte filling. Since high-fat diets facilitate adipocyte enlargement, the early "breakpoint" of hyperphagia seen with the high-fat diet may indicate that this feeding stimulation decreases as the fat cells of the obese rat approach maximal size.     

Increased carbohydrate intake as a function of insulin administration in rats

Patterns of dietary self-selection were examined in male rats following subcutaneous injections of NPH insulin. Self-selection animals received separate dietary sources of protein, carbohydrate and fat, while single diet animals received ground Purina Rodent Chow. Food intakes and body weights were measured daily. In comparison to non-injected animals, both self-selection and single-diet animals displayed similar increases in caloric intakes and body weights in response to insulin administration. In animals maintained on the self-selection regime, the increase in caloric intake was solely a function of an increase in carbohydrate intake. Neither protein intake nor fat intake varied as a function of insulin administration. While fat intake remained stable in insulin-injected animals, non-injected self-selection animals gradually increased fat consumption throughout the experiment. Termination of insulin administration for all animals led to decreases in caloric intake below the levels of non-injected animals. Again, modification in caloric intakes in selecting animals were primarily a consequence of altered carbohydrate intake. Patterns of dietary self-selection in insulin-injected animals are contrasted to patterns of selection in other forms of experimental obesity.     

The effects of continuous naltrexone infusions on diet preferences are modulated by adaptation to the diets.

Two groups of male rats were placed on a feeding regimen in which a fat/protein diet and a carbohydrate/protein diet were available ad lib. Naltrexone was infused via osmotic minipumps either at the time the diets were introduced or after one week of adaptation to the diets. In rats adapted to the diets, naltrexone caused a decrease in the intakes of fat/protein and carbohydrate/protein diets. Relative preferences for the two diets were generally unchanged. In contrast, when naltrexone was infused at the time of introduction of the diets, a polarization phenomenon was observed: rats tended to consume nearly all of their daily calories from either one diet or the other. Six rats (out of 10) showed a stronger preference for the carbohydrate/protein diet than did any of the saline-treated rats, while 3 showed a stronger preference for the fat/protein diet than did any of the saline-treated rats. Thus, the effect was not diet- or macronutrient-specific. These preferences became significantly less extreme after termination of naltrexone infusions. Conditioned aversions and naltrexone-induced reductions in exploratory behavior are discussed as potential explanations for this polarization effect. These results indicate that naltrexone has differential effects on the development versus the maintenance of diet preferences. Further, they emphasize the importance of examining individual differences as well as baseline preferences in studies on the control of intake and diet selection.     

Caloric compensation following insulin administration in rats

Caloric intakes following injections of insulin were examined in male rats maintained on diets of different caloric densities. Following saline or insulin injections, food intake was measured every two hours for six hours or food was withheld for six hours until recovery of normo-glycemia and total intake was measured at seven and eight hours post-injection. Additionally, levels of plasma metabolic fuels, liver glycogen and stomach contents were measured following saline and insulin injections. Animals in both the continuous and delayed access conditions consumed the same number of calories following injections of insulin. Similarly, the increases in intake relative to saline baseline conditions were comparable across diet groups in both conditions although animals tested in the delayed access paradigm were less variable in their response. Changes in levels of metabolic fuels, stomach contents and liver glycogen were found to be consistent with the known effects of insulin and counter-regulatory responses to the induced hypoglycemia.     

Nutrient composition: effects on appetite in monkeys with oral factors held constant

The effects of macronutrients on appetite and total caloric intake in monkeys (Macaca mulatta) was studied using a new feeding and infusion system which yoked intragastric infusion of various nutrients to oral ad lib intake and removed the confounding factor of palatability from the assessment of nutrient effects on feeding behavior. A suction-activated liquid diet feeding system provided free access to a nutritionally complete diet, with 1 ml of diet delivered orally by pump with each discrete suck by the monkey. A second pump was yoked to the oral feeding pump and delivered various nutrients directly into the stomach via an implanted intragastric cannula. Thus, while oral diet composition remained constant, the net diet reaching the stomach varied over ranges of 28 to 77% carbohydrate, 16 to 65% fat and 7 to 36% protein. No significant differences in total caloric intake were observed between intakes of diets with net composition of high carbohydrate or high fat. When protein was increased to 36%, total caloric intake was generally reduced, and this effect was sustained for at least 3 weeks. Therefore, protein appears to have an increased specific satiating effect beyond the caloric content, when compared to carbohydrate or fat.