The effect of weight loss on protein profiles of gastrocnemius muscle in rabbits: a study using 1D electrophoresis and peptide mass fingerprinting

The study of physiological changes occurring during selection contributes to an improved understanding of relationships leading to efficiencies in animal production. To investigate the effects of food restriction in gastrocnemius muscle protein expression, 20% weight reduction was induced in New Zealand White (meat producing) and wild rabbits, using one‐dimensional gel electrophoresis and peptide mass fingerprinting. Lower expression levels of myosin heavy chains were found in the Wild Rabbits Restricted Group, while myosin light chain and α‐crystallin proteins were not detected in restricted groups. Glyceraldeyde‐3‐phosphate dehydrogenase and glycogen phosphorylase expression levels were similar for all experimental groups. Phosphopyruvate hydratase β was not detected in the wild rabbit restricted diet group. Pyruvate kinase levels were 50% lower in the New Zealand Restricted group. LIM protein detection was absent in the control New Zealand group. Results also show relevance of actin in preserving muscle structure in depressed food availability, the sensitivity of both myosin light chain and α‐crystallin protein to restricted feed and the role of PK in the resistance of New Zealand rabbits to food restriction.     

The effect of low‐protein grower diets on the subsequent response of pullets to quantitative food restriction during lay

1. The laying responses of pullets fed from 8 weeks of age on a 12.5% protein grower diet and subsequently subjected to food restriction commencing at various times between 20 and 40 weeks of age were compared with those of pullets reared on diets containing 12.5 or 16% protein and given access to food throughout lay.

2. Restricted feeding during lay of pullets reared on the low‐protein diet substantially improved laying performance, the best results being obtained from pullets fed the low‐protein grower diet and restricted from 20 weeks. The laying responses of pullets restricted after peak‐lay at 40 weeks of age were not significantly different from those of birds restricted prior to peak‐lay.

3. Maximal egg production of 81 % over the 48weeks of the experiment was attained on an average daily ME intake of 270 kcal (1130 kJ) and a FCR of 2.06 : 1.

4. It is suggested that food restriction of laying pullets previously reared on low‐protein diets may be successfully utilised prior to peak‐lay provided such restriction is not so severe as to retard the rate of attainment of mature weight and a continued slight gain in weight thereafter.     

The influence of folic acid, vitamins B(2) and B(6) supplementation on feed intake, body and organs weight, and liver fatty acids composition of rats subjected to 3 months moderate protein deprivation

This study was conducted to determine the effect of a 3-month dietary protein restriction - protein provided 9% of energy (20% in control group). In this dietary restriction folic acid, vitamins B(2) and B(6) were delivered in amount three times above the standard level. It was observed that animals fed a protein restricted (PR) diet weighed about 5% less than animals consuming adequate diet, but the difference was not statistically significant. Enrichment of PR diet with vitamin B or folic acid caused tendency to further suppression of weight gain, and in case of vitamin B(6) these differences were statistically significant. However, such body weight (BW) suppression was not observed when all studied vitamins were used together. Significant reductions in relative liver weight (vitamin B(2) addition), the heart (folic acid) and the lungs (vitamin B(6)) were observed. The PR diet, when all vitamins were added together, caused a decrease in weights of the lungs, heart and liver scaled to BW of rats, simultaneously with a significant increase in testis weight. Feed intake and feed conversion ratio were higher in animals given PR diet without a significant influence of vitamin supplementation (except vitamin B(6) causing further increase in feed conversion ratio). Hepatic fatty acids composition of rats was not affected by protein restriction, as well as by single vitamin supplementation. However, dietary supplementation of all examined vitamins together caused a decrease in monounsaturated fatty acids followed by an increase in polyunsaturated fatty acids participation in total fatty acids pool. It seems that enrichment of PR diet with a mixture of folic acid, vitamins B(2) and B(6) resulted in a partial reverse of growth suppression and reduction in testis size in rats.     

A quantitative analysis of the literature concerning the restricted feeding of growing pullets

The sexual maturity of growing pullets can be retarded either by restricting calorie intake or by feeding a diet deficient in one or more essential amino acids, Several methods have been proposed for achieving either of these objectives. In this review the methods have been classified as (a) limiting the birds’ time of access to food, (b) quantitative food restriction, (c) the use of low energy diets and (d) dietary protein restriction.

Food restriction by limiting the time of access to food has given inconsistent results and is an unsatisfactory technique. Birds quickly learn to eat normal quantities of food in a very short space of time.

Quantitative food restriction has been widely used in experiments and is usually applied from about 6 to about 20 weeks of age. The available evidence enables relationships to be established between the degree of food restriction and subsequent body weight and sexual maturity. The effect of rearing treatment on mortality and rate of lay is also examined.

The ad libitum feeding of a low‐energy growing diet has always resulted in greater rearing food costs. This is because moderate reductions in energy content of the diet are offset by higher food intake. Diets sufficiently low in energy to cause a reduction in calorie intake will involve the purchase and handling of very large proportions of non‐nutritive material.

The use of protein‐deficient diets during the growing period, or from one day of age, offers a suitable means of delaying sexual maturity. The effects of such a treatment are apparently very similar to those of restricted feeding, but there is a lack of suitable comparisons of the alternative methods.     

Welfare assessment of broiler breeders that are food restricted after peak rate of lay

1. Broiler breeder females were reared on one of three growth curves (ad libitum, conventional or modified restriction) and given rations containing a high or low concentration of crude protein. After the peak rate of lay they were fed ad libitum or a decreasing quantity of food in response to declining egg production. The welfare of the birds was assessed by determining the changes in indices of welfare at 36, 48 and 60 weeks of age. 2. Body weight increased rapidly in restricted birds fed ad libitum post-peak and water intake declined. 3. Post-peak food restriction was associated with a decrease in resting and increased drinking and spot-pecking activities. Birds that were food restricted during rearing spent more time foraging and spot-pecking at 36 and 48 but not 60 weeks of age. 4. Immune function increased with age but was not affected by the experimental treatments. The heterophil-lymphocyte ratio in birds fed ad libitum during rearing was numerically lower at 36 and higher at 48 and 60 weeks of age compared with restricted birds. 5. There was no effect of treatment on plasma corticosterone concentration. Creatine kinase activity was high at 60 weeks in treatments that were characterised by poor reproductive status and the activities of other enzymes reflected differences in reproductive status and mortality. 6. There was no long-term welfare or production advantage from feeding low protein rations or more generous feeding during the rearing period compared with conventional food restriction programmes.     

Effects of low dietary protein and different allocations of food during rearing and restricted feeding after peak rate of lay on egg production,fertility and hatchability in female broiler breeders

1. A 3x2x2 factorial experiment was conducted with boiler breeder females to determine the effects of body weight gain (three levels) and conventional or lower crude protein concentrations in the rations during rearing, and restricted or ad libitum feeding after the peak rate of lay on egg production, fertility, hatchability, mortality and food consumption. 2. Compared with ad libitum feeding, conventional food restriction resulted in a decrease in average daily food consumption of 0.6 during rearing, 0.2 during early lay and an increase of 0.4 after the peak rate of egg production. Mortality was decreased by more than half. 3. Restricted birds had higher total and settable egg production, fewer defective or damaged eggshells and higher fertility and hatchability than those fed ad libitum. The modified (more generous) rearing programme resulted in lower rates of egg production and higher rates of mortality compared with the conventional food restriction programme. 4. Low-protein rearing rations were associated with higher rates of food intake, higher mortalities and lower rates of egg production than the conventional protein rations. There were no differences in the fertility or hatchability of eggs between birds fed on the two concentrations of dietary crude protein. 5. Ad libitum feeding post-peak was associated with higher rates of mortality to 60 weeks of age. Post-peak feeding had little effect on the rate of lay or egg weight in conventionally restricted birds fed high protein rations or in birds fed ad libitum. Restricted feeding post-peak decreased the rate of lay and egg weight in birds on the modified restriction programme and in conventionally restricted birds fed on the low protein rations. There was no effect of post-peak feeding on fertility or hatchability of eggs. 6. The total numbers of saleable chicks per kg food consumed were 1.83, 1.72 and 0.52 for conventional, modified and ad libitum feeding during rearing; 1.56 and 1.15 for restricted and ad libitum feeding post-peak: there were no differences associated with protein concentrations of rations fed during rearing.     

AMP-activated protein kinase and adipogenesis in sheep fetal skeletal muscle and 3T3-L1 cells

Marbling, or i.m. fat, is an important factor determining beef quality. Both adipogenesis and hypertrophy of existing adipocytes contribute to enhanced marbling. We hypothesized that the fetal stage is important for the formation of i.m. adipocytes and that AMP-activated protein kinase (AMPK) has a key role in adipogenesis during this stage. The objective of this study was to assess the role of AMPK in adipogenesis in fetal sheep muscle and 3T3-L1 cells. Nonpregnant ewes were randomly assigned to a control (Con, 100% of NRC recommendations, n = 7) or overfed (OF, 150% of NRC, n = 7) diet from 60 d before to 75 d after conception, when the ewes were killed. The fetal LM was collected at necropsy for biochemical analyses. The activity of AMPK was less in the fetal muscle of OF sheep. The expression of peroxisome proliferator-activated receptor (PPAR)gamma, a marker of adipogenesis, was greater in OF fetal muscle compared with Con fetal muscle. To further show the role of AMPK in adipogenesis, we used 3T3-L1 cells. The 3T3-L1 cells were incubated in a standard adipogenic medium for 24 h and 10 d. Activation of AMPK by 5-aminoimidazole-4-car-boxamide-1-beta-d-ribonucleoside dramatically inhibited the expression of PPARgamma and reduced the presence of adipocytes after 10 d of differentiation. Inhibition of AMPK by compound C enhanced the expression of PPARgamma. In conclusion, these data show that AMPK activity is inversely related to adipogenesis in fetal sheep muscle and 3T3-L1 cells.     

Impact of Food Restriction on Ovarian Development,RFamide‐Related Peptide‐3 and the Hypothalamic–Pituitary–Ovarian Axis in Pre‐Pubertal Ewes

RFamide‐related peptide‐3 (RFRP‐3), the mammalian ortholog of gonadotropin‐inhibiting hormone, has been implicated as a mediator between reproduction and energy balance. This study aimed to investigate the physiological effects of RFRP‐3 on the process of ovarian development in food‐restricted pre‐pubertal ewes. The results showed that food restriction significantly inhibited the ovarian development and follicular growth. The data of qPCR in the hypothalamic–pituitary–ovarian (HPO) axis showed that food restriction not only upregulated RFRP‐3 mRNA expression but also downregulated the mRNA expression of gonadotropin‐releasing‐hormone receptor, follicle‐stimulating hormone receptor and luteinizing hormone receptor (LHR). Immunohistochemistry of RFRP‐3 in the ovaries suggested that RFRP‐3 may regulate the follicular development. These results suggested that the changes of RFRP‐3 in response to food restriction might influence the HPO axis and inhibit ovarian development.     

Differential leucocyte responses to various degrees of food restriction in broilers, turkeys and ducks.

1. White blood cell responses of broilers, turkeys and ducks were examined at regular intervals after being subjected to various degrees of food restriction. 2. Restricted-fed broilers showed increases in heterophil and basophil numbers, together with a corresponding decrease in lymphocytes. The heterophil/lymphocyte ratio was raised. There were no differences between broiler strains. 3. After only one week of feeding restricted diets, heterophils were significantly raised in selected and unselected 2-week-old ducks. At 21 weeks of age, those ducks receiving 50% of food required to achieve their ad libitum-fed body weight had raised heterophils. 4. Ducks receiving food to achieve 25% of ad libitum-fed birds produced a marked basophilia, but no heterophilia. 5. After two weeks of food restriction, turkeys responded with significant heterophil/lymphocyte ratios following two degrees of restricted feeding. 6. It was concluded that in some poultry, a heterophilia may be the response to mild to moderate stress but a basophilia may result after severely stressing birds.     

Effect of method and phase of nutrient restriction during rearing on productive performance of light hybrid pullets and hens

1. An experiment was conducted to study the effect of method (low-lysine or quantity of food) and phase (starter or grower) of restriction during rearing on growth and laying performance in Leghorn hybrids. 2. A reduction in body weight gain was achieved by feeding pullets a low-lysine diet in either the starter phase (0-6 weeks of age: 4.0 g/kg digestible lysine) or the grower phase (7-18 weeks of age: 3.0 g/kg digestible lysine). 3. In each phase, the lysine restriction was compared with a daily food restriction, in which pullets received a quantity of food sufficient to pair-gain with their lysine-restricted counterparts. 4. The delay in onset of production was greatest in the lysine-restricted birds, for both phases of restriction. 5. Hens restricted during the grower phase laid heavier eggs than those restricted during the starter phase, despite having similar body weights at end of rearing. This effect was not influenced by the restriction method. 6. Production traits varied between pullets reared on different feeding strategies up to similar body weights at 18 weeks and onwards. It is concluded that body weight is not the only factor determining laying performance.     

Effect of transportation and pre‐slaughter water shower spray with resting on AMP‐activated protein kinase,glycolysis and meat quality of broilers during summer

The aim of this study was to determine the effects of pre‐slaughter transport during summer and subsequent water shower spray on stress, postmortem glycolysis, energy metabolism and adenosine monophosphate‐activated protein kinase (AMPK) in Pectoralis major (PM) muscle of broilers. Results indicated that transport during high ambient temperature induced the release of plasma corticosterone, which significantly affected stress conditions. Moreover, we found a lower energy status in the early postmortem period compared to the control group. AMPK was activated in this situation, following by the rapid glycolysis and accumulation of lactic acid, leading to a high incidence of pale, soft, exudative (PSE)‐like meat. Water shower spray with resting after transport relieved the stress situation, recovered energy homeostasis and lessened the deterioration of meat quality. As a key molecular target for the control of energy status, AMPK has a similar potential in regulating postmortem muscle glycolysis of broilers as in mammals.     

Effects of diet restriction on life span and age-related changes in dogs

OBJECTIVE: To evaluate the effects of 25% diet restriction on life span of dogs and on markers of aging. DESIGN: Paired feeding study. ANIMALS: 48 Labrador Retrievers. PROCEDURES: Dogs were paired, and 1 dog in each pair was fed 25% less food than its pair-mate from 8 weeks of age until death. Serum biochemical analyses were performed, body condition was scored, and body composition was measured annually until 12 years of age. Age at onset of chronic disease and median (age when 50% of the dogs were deceased) and maximum (age when 90% of the dogs were deceased) life spans were evaluated. RESULTS: Compared with control dogs, food-restricted dogs weighed less and had lower body fat content and lower serum triglycerides, triiodothyronine, insulin, and glucose concentrations. Median life span was significantly longer for dogs in which food was restricted. The onset of clinical signs of chronic disease generally was delayed for food-restricted dogs. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that 25% restriction in food intake increased median life span and delayed the onset of signs of chronic disease in these dogs.     

Anticancer effects of resveratrol in canine hemangiosarcoma cell lines

Hemangiosarcoma (HSA) is a highly malignant tumour with aggressive biological behaviour. HSAs are more common in dogs than other domestic animals. The median survival time of dogs with HSA remains short, even with chemotherapy and surgery. Therefore, there is a critical need to improve the adjuvant chemotherapeutic regimens to improve clinical outcomes in dogs with HSA. Resveratrol has been shown to possess strong anti‐proliferative and/or pro‐apoptotic properties in human cancer cell lines. Nevertheless, the potential anticancer effects of resveratrol have not been reported in canine HSAs. The objective of this study is to determine the growth inhibitory effects of resveratrol in HSA cells when used alone or in combination with doxorubicin, a commonly used chemotherapeutic agent. Frog and DD‐1 canine HSA cell lines were treated with varying concentrations of resveratrol with and without doxorubicin. Cell viability was measured by the MTT assay. The expression of apoptotic proteins, activation of p38 mitogen‐activated protein kinase (MAPK), AMP‐activated protein kinase (AMPK) and extracellular signal‐regulated kinase 1/2 (ERK1/2) were assessed by western blotting. Similar to human cancer cell lines, resveratrol markedly inhibited the growth and induced apoptosis in both HSA cell lines. Mechanistically, resveratrol activated p38 MAPK, but did not affect the AMPK or the ERK1/2 pathways. Additional experiments showed that resveratrol augmented the growth‐inhibitory and apoptotic effects of doxorubicin in both HSA cell lines. These findings suggest that resveratrol has pro‐apoptotic effects in canine HSA cells; therefore, its use as a potential adjunct therapy in canine HSA patients warrants further investigation.     

牛AMPK家族基因的研究进展

AMP激活的蛋白激酶（AMP-activated prorein kinase,AMPK）是一种能被腺苷一磷酸（AMP）激活的蛋白激酶。在动物应激（生理、营养、环境和疾病等）过程中起着重要作用。近年研究表明,AMPK在细胞和全身组织器官能量代谢平衡中起着举足轻重的作用,并且在调节物质代谢中起重要作用,其调节代谢过程主要包括葡萄糖转换,糖酵解和脂类代谢等。近年来在牛上已经进行了很多研究,其中包括其染色体定位及结构的确定等。本文将简要介绍AMPK的分子结构及功能,并对AMPK基因在牛上的研究进展进行综述。     

Clinical and metabolic findings in dogs with chronic renal failure fed two diets.

Exogenous creatinine clearance, urinary electrolyte excretions, calcium and phosphorus balance, serum cholesterol concentration, arterial blood pressure, and body weight were evaluated in dogs with chronic renal failure that were fed 2 commercial diets. Nine dogs ranging in age from 1 to 15 years were identified as having mild to moderate chronic renal failure (CRF, exogenous creatinine clearance = 0.5 to 2.13 ml/kg of body weight/min). These dogs and a group of 10 clinically normal controls were fed a diet containing 31% protein for 8 weeks at which time hematologic and biochemical evaluations and clearance studies were performed. All dogs then were fed a phosphorus-restricted diet containing 16% protein and then reevaluated after 8 weeks. The dogs in this study had hematologic and biochemical abnormalities typical of CRF. Urine absolute and fractional excretion of electrolytes was higher in dogs with CRF than in controls and was affected by diet. Serum cholesterol concentration was higher in dogs with CRF and increased in those dogs after feeding the low protein diet. Changes in dietary sodium intake did not affect arterial blood pressure. The phosphorus-restricted diet did not affect serum amino terminal parathyroid hormone concentration in either group. Control dogs lost body weight, whereas dogs with CRF gained weight when fed the low protein diet. We concluded that dogs with mild to moderately severe CRF have the same biochemical abnormalities and response to dietary restriction of protein and phosphorus as has been previously reported in dogs with experimentally induced CRF. Restriction of dietary sodium may not decrease arterial blood pressure in some dogs with CRF.(ABSTRACT TRUNCATED AT 250 WORDS)     

Arginine--lysine interrelationships in the nutrition of broiler breeder pullets during the developmental period

Pullets were fed on diets containing various concentrations of lysine and arginine from 7 to 18 or 20 weeks. Thereafter a conventional layers' diet was fed and egg productivity measured—in the first two experiments for ten, and the third for six 28‐d periods. Pullets were also fed on a standard growers' type diet on a “skip‐a‐day” or on a 20% restriction basis in the third study.

Growth and age at sexual maturation were directly related to the lysine content of the diet and to a lesser extent to the arginine content. Pullets fed on diets with the lower concentrations of lysine and arginine required less food during the growing period and matured later than those fed on a complete diet ad libitum. They also laid more eggs and required less food to produce a dozen eggs. Higher egg production was obtained in the third experiment with low concentrations of lysine and arginine than with the “skip‐a‐day” or the restricted feeding programmes. Pullets fed on the normal grower diet ad libitum produced the smallest eggs in the first and third experiments with no difference in egg size in the second experiment. Pullets maintained on the “skip‐a‐day” or the restricted feeding programme also had a larger egg size than the control group fed on the same diet on an ad libitum basis.

An ingredient effect was observed in the third experiment. Of two diets containing identical calculated values of lysine and arginine, the one containing distillers' dried grains with solubles was superior to the one with soybean meal as regards weight at 20 weeks of age, amount of food consumed during the period 6 to 18 weeks of age and percentage egg production. It is possible that the soybean meal contained a better balance of amino acids than did distillers' dried grains with solubles which resulted in a significant increase in body weight at 20 weeks of age and a subsequent lower level of egg production. These data indicate that it would be best to use ingredients with an imbalance of amino acids in the protein as sources of lysine and arginine in pullet grower diets.

There were no differences in the mortality of the birds during the laying period which could be related to dietary treatment during the pullet growing period.     

Use of nonprotein nitrogen in pigs: effects of dietary urea on the intestinal microflora

The fecal flora of 2 pigs given an optimal protein diet (digestive crude protein, 13.3%) with 2% urea, 2 pigs given a low protein diet (digestive crude protein, 5.7%) with 2% urea, and 2 pigs given a low protein diet only were bacteriologically investigated before feeding of each diet and on feeding days 21 and 38. The addition of urea to a low protein diet was significantly decreased Lactobacillus (P less than 0.001), Bacteroidaceae (P less than 0.01), Peptococcaceae (P less than 0.05), and Megasphaera (P less than 0.01) in the feces. In addition, there was a marked suppression of anaerobic curved rods (mainly Selenomonas) to undetectable amounts in the feces. A significant reduction (P less than 0.01) of Bifidobacterium and a marked suppression of Megasphaera to undetectable amounts were observed in the feces of pigs given low protein diets without urea. Microbial groups in the ileum and colon of pigs on feeding day 38 were also analyzed. Effects of dietary urea on the intestinal flora included: (i) anaerobic curved rods and spiral rods were restricted mainly to the colon, (ii) smaller numbers of yeasts were present in the intestine, and (iii) Bacteroidaceae, Peptococcaceae, and Eubacterium in the colon were not changed. Many anaerobic isolates, including curved rods and spiral rods, in the colon of pigs may have an important role in urea use.     

Effect of interaction between leucine and acetate on the milk protein synthesis in bovine mammary epithelial cells

The interaction between Leucine (Leu) and acetate affecting milk protein synthesis in the bovine mammary epithelial cells (BMECs), and underlying the molecular mechanisms are not well understood. The objectives of this study were to investigate the effect of Leu, acetate, and their interaction on the expression of genes involved in milk protein synthesis, and JACK2/STAT5, mTOR and AMP‐activated protein kinase (AMPK) signaling pathway. The study was a 2 × 6 factorial arrangement with treatments: Leu concentration (0.45 and 1.8 mM) and acetate concentration (0, 4, 6, 8, 10, and 12 mM). The results showed that 1.8 mM Leu or 8–10 mM acetate had positive effect on ATP content, the expression of casein genes, JACK2/STAT5 and phosphorylation of mTOR pathway, but reduced AMPK phosphorylation. Leu at 1.8mM had a positive effect on the up‐regulation of acetate on ATP content, the expression of CSN1S1, CSN2, CSN3, and JACK2, the expression and phosphorylation of eukaryotic initiation factor 4E, p70 ribosomal protein S6 kinase‐1, and mTOR, but reducing AMPK phosphorylation. The results suggest that acetate, Leu, and their interaction have effect on milk protein synthesis through the JACK2/STAT5, mTOR, and AMPK pathway. Acetate addition up‐regulated the effect of Leu on milk protein synthesis, and Leu facilitated the up‐regulation of acetate on milk protein synthesis through these pathways.     

低聚果糖诱导的奶牛急性蹄叶炎肝和骨骼肌中AMPK和GLUT及相关基因的变化

旨在探究奶牛急性蹄叶炎能量代谢的变化,本研究选用12头健康中国荷斯坦奶牛,随机分为两组(n=6): 试验组(OF组)用17 g·kg^-1体重的低聚果糖溶解在2 L·100 kg^-1体重的清水中灌服,对照组(CON组)灌服等量清水,72 h之后对奶牛进行安乐死,采集肝、肌肉组织,进行Western blot试验和实时荧光定量PCR试验,检测肝、肌肉组织的能量变化和葡萄糖转运情况,指标：葡糖糖转运蛋白1和4(GLUT-1、GLUT-4),三磷酸腺苷激酶(AMPK)以及相关因子(PPAR-γ、PGC1-α、PEPCK)。结果表明：在肝组织中,OF组AMPK和蛋白的表达量显著增加,但P-AMPK/AMPK的比值极显著下降,而GLUT-1的蛋白和基因、PPAR-γ、PGC1-α和PEPCK基因的表达量无显著变化;在肌肉组织中,OF组AMPK基因和蛋白表达量无显著的变化,但P-AMPK/AMPK的比值显著下降,GLUT-4基因和蛋白表达量显著下降,同时PPAR-γ、PEPCK基因的表达量极显著升高,但PGC-1-α的基因表达量无显著的变化。综上：奶牛急性蹄叶炎可能会抑制肌肉组织的能量代谢和葡萄糖转运的能力,但对肝的影响不大。