Restricted Intake Groups Research Articles

Effect of early feed restriction programs and genetic strain on humoral immune response production in broiler chickens

Two experiments were conducted to compare the immunocompetence of Cobb high performance and rustic Label Rouge broilers and the influence of reduced growth rates subsequent to feed restriction on the IgY anti-bovine serum albumin (BSA) response. In the first experiment (EXP), 360 broilers were assigned to 36 cages from 1 to 42 days of age. A completely randomized design was applied in a 3 × 2 factorial arrangement, with 3 groups (Label Rouge, Cobb ad libitum, and Cobb Restricted Intake), and 2 levels of energy (3,100 and 2,800kcal/kg); there were 6 replicates per treatment. In the second EXP, 384 Cobb 500 male broilers were randomly assigned to the following feed restriction programs from day 8 to 16: Control, fed ad libitum; Quantitative (80% of the control amount); By Time (fed for 8 h/d), and Qualitative (80% limiting nutrients) restriction. Blood samples were collected on days 35 and 42 (EXP 1) and weekly from day 7 to 42 (EXP 2) for IgY anti-BSA quantification. In EXP 1, the production of IgY anti-BSA was lower in the Cobb groups (P < 0.0001) than in the Label Rouge group, and higher in the Cobb Restricted Intake group (P < 0.0001) compared with the same genetic strain fed ad libitum. Birds fed the low energy diet presented lower (P ≤ 0.06) IgY anti-BSA, independent of genetics. In EXP 2, no difference (P > 0.05) was observed 1 wk after the first BSA inoculation. However, at day 28, birds in all feed restriction programs had higher (P < 0.05) IgY anti-BSA than the Control group fed ad libitum. At day 35, the greatest residual effect of IgY anti-BSA was observed in the Quantitative restriction group. No differences (P > 0.05) were observed between groups after 42 d. The 3 early feed restriction programs had beneficial effects on the humoral immune response. Overall, Quantitative restriction promoted a longer lasting IgY anti-BSA response. Lower growth rate, due to feed restriction or genetic potential, improves humoral immunity in broiler chickens.

Ad Libitum vs. Restricted Fluid Replacement on Hydration and Performance of Military Tasks

The primary objective was to evaluate the effect of ad libitum vs. restricted fluid replacement protocol on hydration markers and performance in selected military tasks. The secondary objective was to determine if 300 ml x h(-1) could be considered a safe minimum fluid intake under the experimental conditions. Data were collected simulating a route march over 16 km. There were 57 subjects who participated in the study. The mean pre-exercise body mass of the ad libitum group was 70.4 +/- 13.3 (SD) kg compared to 69.3 +/- 8.9 kg in the restricted group. The mean total fluid intake of the ad libitum group was 2.1 +/- 0.9 L compared to 1.2 +/- 0.0 L in the restricted group. The ad libitum and restricted intake groups, respectively, lost a mean of 1.05 kg +/- 0.77 (1.5%) and 1.34 kg +/- 0.37 (1.9%). Calculated sweat rate was 608 +/- 93 ml x h(-1) compared to 762 +/- 162 ml x h(-1) in the ad libitum group. There were no significant differences for either urine specific gravity (USG) or urine osmolality (UOsm) before or after the exercise. It is not clear whether fluid intake and calculated sweat rates are causally related or explained by their codependence on a third variable; for example, the exercising metabolic rate. Thus, 300 ml x h(-1) intake could be considered a current safe minimum water intake for soldiers of similar mass under similar experimental conditions, namely similar exercise durations at equivalent exercise intensities in a moderate, dry climate.

Elevated blood interleukin-10 levels and undiminished systemic interleukin-10 production rate prevail throughout acute protein-energy malnutrition in the weanling mouse

The objectives were to determine if blood IL-10 levels rise during the early stages of acute (wasting) pre-pubescent malnutrition in metabolically distinct murine models known to depress inflammatory immune competence and whether systemic IL-10 production is affected in these pathologies. Weanling C57BL/6 J mice were assigned to dietary protocols that elicited wasting pathologies mimicking the human diseases of marasmus (restricted-intake group) or incipient kwashiorkor (low-protein group). An age-matched control group also was included. Serum IL-10 bioactivities were assessed in the early (day 3) and advanced (day 14) stages of weight loss, and net systemic IL-10 production was assessed at the same stages of pathology by in vivo cytokine capture. Blood IL-10 levels were elevated in both malnourished groups relative to controls at days 3 and 14 (range of P values: 0.03–0.0001). Further, despite a limited supply of energy and nitrogenous substrates, the systemic IL-10 production rate was at least sustained in the malnourished groups and, in fact, was elevated in the marasmic group ( P = 0.05) throughout the progression of weight loss. IL-10 emerges as an anti-inflammatory mediator positioned to participate in initiating and upholding the depressed immune competence that accompanies acute pre-pubescent deficits of protein and energy.

Effector/memory T cells of the weanling mouse exhibit Type 2 cytokine polarization in vitro and in vivo in the advanced stages of acute energy deficit

Our objective was to determine whether the polarizing cytokine profile of the effector/memory T-cell compartment reflects the profound decline of cell-mediated inflammatory competence that characterizes acute prepubescent malnutrition. Weanling C57BL/6J mice were permitted free access to a complete purified diet, free access to an isocaloric low-protein diet or restricted intake of the complete diet for 14 days. First, interleukin (IL)-4 and interferon (IFN)-γ concentrations generated in vitro by splenic and nodal effector/memory T cells were assessed following exposure to plate-bound anti-CD3. Second, net systemic production of IFN-γ and IL-4 by the effector/memory T-cell compartment was assessed by the in vivo cytokine capture assay following anti-CD3 stimulation. In vitro stimulation generated less IFN-γ ( P=.002) but more IL-4 ( P=.05) by T cells from the restricted-intake group relative to the age-matched control group. Similarly, in vivo stimulation generated low serum levels of antibody-captured IFN-γ in the restricted-intake group vis-à-vis the age-matched control group ( P=.01), while the IL-4 response was sustained ( P=.39). By contrast, the 14-day low-protein model exhibited no change in T-cell cytokine signature either in vitro or in vivo. However, following extended consumption of the low-protein diet (26 days), carcass energy losses exceeded those of the 14-day protocol and serum levels of in vivo antibody-captured IFN-γ were low after anti-CD3 challenge relative to the age-matched control group ( P=.02), while levels of captured IL-4 remained unaffected ( P=.07). Acute weanling malnutrition elicits a Type 2 polarizing cytokine character on the part of the effector/memory T-cell compartment, but only in the most advanced stages of energy decrement.

Blood serum interferon-γ bioactivity is low in weanling mice subjected to acute deficits of energy or both protein and energy

The main objective of the present study was to determine the influence of acute deficits of protein and energy on the blood serum level of interferon-gamma, a signature type 1 polarising inflammatory cytokine. In two 14 d experiments, male and female C57BL/6J mice, initial age 19 d, consumed a complete purified diet ad libitum or in restricted daily quantities, or had free access to an isoenergetic purified low-protein diet. A zero-time control group (age 19 d) was included in the second experiment. Serum interferon-gamma was assessed in both experiments by sandwich ELISA and, in the second experiment, also by a bioassay based on inhibition of proliferation by WEHI-279 B lymphoma cells. The immunoassay detected interferon-gamma inconsistently in all groups (range 0-14 pg/ml; detection limits 1 x 5 and 0 x 7 pg/ml in experiments 1 and 2, respectively). By contrast, interferon-gamma bioactivity was found in all animals of each group (means 339, 499, 124 and 200 pg/ml in zero-time controls, age-matched controls, low-protein and restricted intake groups, respectively; detection limit, 12 pg/ml), and the mean serum bioactivity of each malnourished group was low compared with the age-matched control (P <or=0 x 05). The present study defines the physiological serum interferon-gamma bioactivity of the adolescent mouse. Moreover, to the extent achievable by way of the blood, the results reflect the influence of metabolically diverse forms of acute malnutrition on the polarising type 1 cytokine profile within lymphoid microenvironments wherein immune responses arise. Therefore, the results suggest a mechanism underlying the cell-mediated inflammatory incompetence that characterises acute, prepubescent malnutrition.

Elevated Bioactivity of the Tolerogenic Cytokines, Interleukin-10 and Transforming Growth Factor-β, in the Blood of Acutely Malnourished Weanling Mice

The main objective of this investigation was to determine the influence of acute deficits of protein and energy on the blood levels of interleukin-10 (IL-10) and transforming growth factor-beta (TGF-beta), physiologically the main anti-inflammatory and tolerogenic cytokines. In four 14-day experiments, male and female C57BL/6J mice, initially 19 days old, consumed a complete purified diet either ad libitum or in restricted daily quantities, or had free access to an isocaloric purified low-protein diet. A zero-time control group (19 days old) was included. In the first two experiments, serum IL-10 levels were assessed by sandwich enzyme-linked immunosorbent assay (ELISA) and bioassay. The mean serum IL-10 bioactivities were higher (P < or = 0.05) in both malnourished groups (low-protein and restricted intake: 15.8 and 12.2 ng/ml, respectively) than in the zero-time and age-matched control groups (6.3 and 7.3 ng/ml, respectively), whereas serum IL-10 immunoactivity was high only in the restricted intake group (e.g., second experiment: 17.0 pg/ml vs. 5.4, 3.7, and 3.1 pg/ml in the zero-time control, age-matched control and low-protein group, respectively). The third and fourth experiments centered on plasma TGF-beta immunoactivity (sandwich ELISA) and bioactivity, respectively. The ELISA revealed a high mean plasma TGF-beta1 level (P < or = 0.05) in the low-protein group only, but TGF-beta bioactivity (beta1 isoform, although 15% beta2 in the restricted intake group) was high in both malnourished groups (8.7 and 9.3 ng/ml in the low-protein and restricted groups, respectively) relative to the age-matched control group (0.5 ng/ml). Thus, metabolically distinct weanling systems mimicking marasmus and incipient kwashiorkor both exhibit a blood cytokine profile that points to a tolerogenic microenvironment within immune response compartments. A model emerges in which malnutrition-associated immune competence, at least in advanced weight loss, centers on cytokine-mediated peripheral tolerance that reduces the risk of catabolically induced autoimmune disease, but this is at the cost of attenuated responsiveness to infectious agents.

Interactions between Growth Hormone and Nutrition in Hypophysectomized Rats: Skeletal Muscle Myosin Heavy Chain mRNA Levels

The aim of this study was to examine the role of growth hormone (GH) in regulating muscle phenotype and to determine how this is modulated by altered nutrition. Total RNA was extracted from gastrocnemius muscles of hypophysectomised rats treated with saline, GH or GH but fed a restricted intake. Type 1, 2A, 2B, embryonic and neonatal myosin heavy chain mRNA levels were estimated by slot blot hybridization. Hypophysectomy reduced the concentrations of types 1, 2A and embryonic mRNAs and dramatically elevated types 2B and neonatal compared to control levels, but this was time-dependant. All MHC mRNA levels were partially restored to control levels in the GH-treated rats except for type 1; the level of this transcript was only elevated by GH in the restricted intake group. Restricted food intake modulated the effects of GH administration for all other MHC mRNA concentrations.

Polyamine-directed preferential nutritional repletion of normal tissues in tumor-bearing hosts.

Our previous study demonstrated that total parenteral nutrition (TPN) increases erythrocyte (RBC) polyamines selectively in cancer patients but not in non-cancer patients, suggesting that these changes may relate to tumor presence. We therefore studied the possible relationship between RBC polyamine levels and potential tumor growth induced by TPN. In addition, we attempted to control tumor growth during TPN with alpha-difluoromethylornithine (DFMO), a suicide inhibitor of ornithine decarboxylase (ODC), in a fibrosarcoma-bearing rat model. Rats in a control group received chow ad libitum for 3 weeks and their tumors grew exponentially. A restricted intake (RI) tumor-bearing group received 8 g chow per day per rat; this produced malnourished rats in which tumor growth significantly slowed. The third group (RI----TPN) received the RI diet for 2 weeks and TPN for 1 week, which resulted in an increase in tumor growth. The last group (RI----TPN + DFMO) received a 2-week RI regimen and 1-week TPN plus DFMO regimen, which inhibited tumor growth to the level of the RI group due to the addition of DFMO (1 g/kg/day). Although the addition of TPN to the RI regimen markedly increased tumor ODC activity, as well as tumor putrescine and RBC putrescine levels, the addition of DFMO to the RI----TPN regimen entirely abolished these increases. Despite DFMO treatment, liver ODC activity in RI----TPN + DFMO group was unaffected. The plasma albumin level, an indicator of host nutrition, of the RI----TPN + DFMO group was significantly higher than that of the RI group. Liver, spleen, kidney and total body weights of the RI----TPN or RI----TPN + DFMO group were significantly higher than those of the RI group. These results indicate that TPN can enhance tumor growth in malnourished hosts and that the addition of DFMO to TPN can effectively control this TPN-enhanced tumor growth, leading to preferential nutritional repletion of the host.

The relation between dietary restriction or clenbuterol (a selective beta 2 agonist) treatment on muscle growth and calpain proteinase (EC 3.4.22.17) and calpastatin activities in lambs.

1. Lamb growth trials were designed to modify growth and protein content of muscle by diet and also by beta-agonist treatment, and to correlate any changes to the activities of calpain proteinases (EC 3.4.22.17) and their inhibitor calpastatin. 2. Wether lambs in a control group were fed on a barley-based diet designed to give a growth rate of 350 g/d; a second group was fed on the same diet but the intake was restricted to give an expected gain of 44 g/d; a third group was fed on the same diet as the first group but the diet included 2 mg clenbuterol/kg. At the end of a 6-week trial, longissimus dorsi wet weights were 635 (n6), 377 (n4) and 788 g (n6) (standard error of difference 53.0) in the three groups respectively. 3. Minced L. dorsi was extracted in low-salt buffers and analysed by a fast protein liquid-chromatographic system for calpain I (low calcium-requiring), calpain II (high Ca2+-requiring) and calpastatin activities. No significant changes in the three activities were associated with reduced muscle weight in the restricted-intake group. The inclusion of clenbuterol in the diet, however, led to highly significant increases (P less than 0.001) in calpain II and calpastatin to approximately double the control values. 4. The results did not support a direct relation between these activities and muscle growth, except when protein accretion was stimulated by a beta-agonist, suggesting a role for this enzyme system in the mechanism by which these agents exert their effect.

Metabolic effects of a “carbohydrate-free” diet in the pregnant rat

Abstrac The effects on selected metabolic indicators of carbohydrate deprivation in pregnant rats was studied. Pregnant rats were fed a diet containing 13% casein, either 69.5% (CON) or 0% (CD) carbohydrate, and oleic acid to make the diets isocaloric. Diets were fed beginning on Day 0 of pregnancy. A restricted intake group (RI) was also included to indicate the effect of reduced food intake. On days 8, 10, and 12 of pregnancy, animals were killed and liver, uterus and blood were harvested. Glycogen and glucose-6-phosphatase in endometrial tissue, glycogen in liver, ketone bodies in whole blood, and glucose, total lipid, cholesterol, urea nitrogen and alanine in plasma were determined. In CD dams, fetal resorption reached 75% by day 12 of gestation but only 35% in RI dams. Carbohydrate deprivation and restricted intake resulted in depleted liver glycogen. CD dams were hypoglycemic and hyperlipemic. Plasma alanine did not differ among the 3 groups and plasma urea was elevated in the CD group only on day 12. CD dams were markedly hyperketonemic. Neither uterine glycogen nor uterine glucose-6-phosphatase activity differed between the 3 groups, suggesting that uterine glycogen utilization was not the limiting factor in embryonic survival. Hypoglycemia, hyperketonemia or both may be related to the reproductive failure in CD rats.

Studies of Nutrition and Autoimmunity. Failure of Zinc Deprivation to Alter Autoantibody Production when Initiated in Disease-Established Mice

Earlier studies have demonstrated that zinc deprivation, when begun in young NZB mice, can retard the development of autoimmunity and result in an increased life span. The present study evaluated the possible benefits of zinc deprivation in NZB mice with established disease, as this model is more relevant to the human patient. Female NZB mice aged 6–8 mo were fed diets containing either 80 μg Zn/g and 10 μg Cu/g (control) or 1 μg Zn/g and 10 μg Cu/g (zinc-deficient) for 4 mo. In addition, another group was fed a diet containing 1 μg Zn/g and 100 μg Cu/g to determine whether zinc deficiency could be exacerbated by high dietary copper through a competition of copper with zinc at the intestinal level (zinc deficient + high copper). A fourth group of mice was fed the control diet at the same intake as that of the zinc-deficient group in order to control for the inanition associated with zinc deficiency (restricted intake). Regardless of dietary treatment, all mice developed antierythrocyte antibodies at the same rate. At the end of 4 mo, 82% of the control and the restricted-intake groups had survived, whereas the zinc-deficient group had a 38% survival rate and the zinc-deficient + high copper group had a 50% survival rate. These observations show that, in contrast to findings with younger NZB mice, zinc deprivation of adult NZB mice with established autoimmunity will not improve survival. Indeed, severe zinc deficiency increased the mortality rate, demonstrating the need to consider the potential hazards of dietary extremes.

Similar Effects of Zinc Deficiency and Restricted Feeding on Plasma Lipids and Lipoproteins in Rats

The objective of this study was to determine whether alterations in HDL composition observed previously were due specifically to zinc deficiency or to reduction in food intake. Rats were placed into three groups and fed for 21 d a zinc-adequate diet (100 ppm) ad libitum, or a zinc-deficient diet (< 1 ppm) ad libitum, or the zinc-adequate diet at an intake equal to that of the deficient group. Blood plasma was fractionated into VLDL, LDL and HDL fractions, and plasma and the isolated fractions were analyzed for lipids and apolipoprotein composition. Analysis of plasma and liver zinc indicated that animals consuming the low zinc diet were deficient. Zinc deficiency and restricted feeding were associated with lower plasma and VLDL triglyceride levels and lower HDL cholesterol levels. The similarities in overall pattern of lipoprotein composition between the zinc-deficient and restricted-intake groups indicate that reduction in food intake may account for the differences in lipoprotein composition which were observed.

Effect of Zinc Status on Salivary Zinc Concentrations in the Rat

Three experiments were done to determine whether salivary zinc concentration is a more sensitive indicator of zinc status than plasma zinc. Weanling male rats fed a low zinc (less than 1 ppm) diet for 5 weeks with or without zinc (100 ppm) in the drinking water had salivary zinc concentration of 0.19, 0.16, and 0.20 microgram/ml for the zinc-deficient, zinc-supplemented restricted-intake, and zinc-supplemented ad libitum-fed groups, respectively. Combined values for male and female rats after 4 weeks of the same treatments in experiment 2 were 0.60, 1.2 and 0.44 microgram/ml. Saliva collected on day 22 of pregnancy contained 0.30 and 0.24 microgram/ml from zinc-supplemented and zinc-deficient rats, respectively. Salivary zinc concentrations in the deficient rats did not differ from those of the zinc-supplemented ad libitum-fed controls in any of the experiments. Salivary zinc concentration in the zinc-supplemented restricted-intake group in experiment 2 was significantly higher than that in the other two groups. Decreases in serum, bone, and fetal zinc concentrations indicated that the rats were definitely zinc-deficient. Since zinc concentration of mixed saliva in the rat was not decreased by even a severe zinc deficiency, salivary zinc does not appear to be as good an indicator of zinc status as plasma zinc.