The present study evaluated the effect of different levels of energy restriction on metabolic parameters in obese ponies. Relative weight changes, markers of lipid metabolism and oxidant/antioxidant balance were monitored. A total of eighteen obese (body condition score ≥ 7/9) Shetland ponies were studied over a 23·5-week trial, which was divided into three periods. The first period involved a 4-week adaptation period in which each animal was fed 100 % of their maintenance energy requirements needed to maintain a stable obese body weight (MERob). This was followed by a 16·5-week weight-loss period in which ponies were assigned to receive either 100 % (control group, CONTROL), 80 % (slow weight-loss (SLOW) group) or 60 % (rapid weight-loss (RAPID) group) of their MERob. During the 3-week end-phase period, all ponies were again fed 100 % of their MERob. Relative weight loss was higher in the RAPID group (P< 0·001) compared with the SLOW group. No linear relationship was found as a doubling of the percentage of energy restriction was accompanied by a tripling of the percentage of weight loss. Relative weight gain afterwards in the end-phase period was higher in the RAPID group (P< 0·001) compared with the SLOW and CONTROL groups. During the weight-loss period, TAG and NEFA concentrations were highest in the RAPID group, as were α-tocopherol and ferric-reducing ability of plasma concentrations. After 8 weeks of weight loss, the concentrations of advanced oxidation protein products were higher in the RAPID group compared with the SLOW and CONTROL groups (P< 0·001). In conclusion, the level of energy restriction influences the extent of changes in oxidant/antioxidant balance. Practically, more severe energy restriction regimens may be associated with a greater regain of weight after the restriction period.

Albumen was removed from broiler eggs before the start of incubation to induce prenatal protein under-nutrition in chicken embryos. With this method, the direct effect of protein deficiency was investigated, differing from mammalian models manipulating the maternal diet where indirect, hormonal effects can interfere. Based on the estimated albumen/egg weight ratio, 10 % of albumen was removed with an 18G needle, after making a hole at the sharp end of the egg with another 18G needle. Eggs were taped thereafter. The sham group underwent the same procedure, except that no albumen was removed. Control eggs did not receive any treatment. The removal of albumen decreased both embryonic and post-hatch body weight up to day 7 compared with the control group. On embryonic day 18, embryos from the albumen-deprived group had higher plasma uric acid levels compared with the sham (P= 0·016) and control (P= 0·009) groups. Moreover, a lower plasma amino acid concentration was observed at hatch compared with the sham (P= 0·038) and control (P= 0·152) groups. These findings indicate an altered protein metabolism. At hatch, a higher mRNA expression of muscle ring finger-1 (MuRF1), a gene related to proteolysis, was observed in albumen-deprived chicks compared with the control and sham chicks, together with an up-regulated expression of atrogin-1 (another atrogene) at this time point in the male protein-deficient chicks. These findings suggest that muscle proteolysis is transiently increased by the removal of albumen before the start of incubation. No evidence was found for altered protein synthesis capacity and translational efficiency in albumen-deprived chicks.

Besides its typical role as an amino acid in protein synthesis, methionine is an important intermediate in methylation reactions. In addition, it can also be converted to cysteine and hence plays a role in the defence against oxidative stress. The present study was conducted to investigate further the role of dl-methionine (DLM) and its hydroxy analogue, dl-2-hydroxy-4-methylthiobutanoic acid (DL-HMTBA), on zootechnical performance and oxidative status of broiler chickens. Male broiler chickens were reared on two diets differing in crude protein (CP) content (low-protein, 18·3 % v. high-protein, 23·2 % CP) and were supplemented either with 0·25 % DLM or 0·25 % DL-HMTBA. Reducing the dietary protein content resulted in an impaired body weight gain (P < 0·0001). However, supplementation of DL-HMTBA to the low-protein diet partially alleviated these negative effects (P = 0·0003). This latter phenomenon could be explained by the fact that chickens fed DL-HMTBA-supplemented diets displayed a better antioxidant status as reflected in lower lipid peroxidation probably as a consequence of their higher hepatic concentrations of total and reduced glutathione compared with their DLM counterparts. On the other hand, within the high protein levels, uric acid might be an important antioxidant to explain the lower lipid peroxidation of high-protein DL-HMTBA-supplemented chickens. Hepatic methionine sulfoxide reductase-A gene expression was not significantly affected by the dietary treatments. In conclusion, the present study indicates that there are interactions between dietary protein content and supplementation of methionine analogues with respect to broiler performance and antioxidant status, also suggesting a causal link between these traits.

N balance and postprandial acylcarnitine profile following intestinal fermentation of oligofructose and inulin were investigated in healthy cats. Two diets were tested in a crossover design: a commercial high-protein cat food supplemented with 4 % DM oligofructose and inulin (spectrum: degree of polymerisation (DP) 2–10: 60 (se 5) % DM; DP>10: 28 (se 5) % DM) as high-fermentable fibre (HFF) diet, and the same commercial diet supplemented with 4 % DM cellulose as low-fermentable fibre diet. Eight adult cats were randomly allotted to each of the two diets at intervals of 4 weeks. At the end of each testing period, faeces and urine were collected over a 5-d period, and blood samples were obtained before and at the selected time points postprandially. No differences were found for N intake, N digestibility and faecal N excretion, whereas urinary N excretion was lower when the HFF diet was fed (P = 0·044). N balance was positive in all the cats, and tended to be increased when the HFF diet was fed (P = 0·079). Propionylcarnitine concentrations (P = 0·015) and their area under the curve (AUC) (P = 0·013) were increased when the HFF diet was fed, revealing a more pronounced production and absorption of propionate. Yet, methylmalonylcarnitine concentrations and concurrent AUC were not elevated when the HFF diet was fed, indicating reduced amino acid catabolism. 3-Hydroxy-3-methylglutarylcarnitine concentrations (P = 0·026) and their AUC (P = 0·028) were also reduced when the HFF diet was fed, implying diminished use of branched-chain amino acids as well. In healthy cats, oligofructose and inulin added to a high-protein diet were suggested to reduce postprandial amino acid-induced gluconeogenesis by substitution with propionate.

The present study assessed the effect of separate reduction of each energy-delivering nutrient – protein, fat and carbohydrate – on glucose tolerance and insulin response in a strict carnivore: the domestic cat (Felis catus). Three isoenergetic, home-made diets with the following energetic distribution, low protein (LP): protein 28 % of metabolisable energy; fat 43 %; nitrogen-free extract 29 %; low fat: 47, 27 and 25 %; low carbohydrate (LC): 45, 48 and 7 %, were tested in a 3 × 3 Latin square design. Nine healthy normal-weight cats were randomly assigned to each of the diets in a random order at intervals of 3 weeks. At the end of each testing period, intravenous glucose tolerance tests were performed. Plasma glucose concentrations and area under the glucose curve showed no differences. Area under the insulin curve was lower when cats were fed the LP diet, and the second insulin peak tended to be delayed when the LC diet was fed. In contrast to other studies, in which energy sources were elevated instead of being reduced, the present trial contradicts the often suggested negative impact of carbohydrates on insulin sensitivity in carnivores, and shows that reducing the dietary carbohydrate content below common amounts for commercial foods evokes an insulin-resistant state, which can be explained by the cats' strict carnivorous nature. It even points to a negative effect of protein on insulin sensitivity, a finding that corresponds with the highly gluconeogenic nature of amino acids in strict carnivores.

The cytoplasmic serine/threonine ribosomal protein S6 kinase (S6K1) plays a critical role in controlling protein translation. There is evidence that amino acids regulate S6K1 and protein synthesis in avian species, but the effect of dietary protein level on the activation of S6K1 in neonatal chicks is unknown. Therefore, the aim of the present experiment was to investigate the effect of different protein levels, supplied during the first 5 d post-hatch, on body growth, breast muscle development and on the activation of S6K1 and its downstream target, the S6, in neonatal chicks. Chicks were fed a pre-starter diet during the first 5 d post-hatch containing low (19·6 % crude protein (CP); LP), medium (23·1 % CP; MP) or high (26·7 % CP) levels (HP) of protein. Weight gain of chicks fed the HP diet was higher (P < 0·05) compared with those fed the LP diet during day (d)3–d5 and the numerical advantage of this group was maintained from d2 to d7. On d2 and d3, greater levels of S6K1 and S6 phosphorylation and/or activity were observed in chicks receiving the HP diet compared with LP and MP diets, without differences between results of the latter two dietary treatments. In conclusion, the present results suggest that early protein nutrition impacts the development of broiler chicks.

The effect of dietary oligofructose and inulin supplementation on glucose metabolism in obese and non-obese cats was assessed. Two diets were tested in a crossover design; a control diet high in protein (46 % on DM basis), moderate in fat (15 %), low in carbohydrates (27 %), but no soluble fibres added; and a prebiotic diet, with 2·5 % of a mixture of oligofructose and inulin added to the control diet. Eight non-obese and eight obese cats were allotted to each of two diets in random order at intervals of 4 weeks. At the end of each testing period, intravenous glucose tolerance tests were performed. Area under the glucose curve (AUCgluc) was increased (P = 0·022) and the second insulin peak was delayed (P = 0·009) in obese compared to non-obese cats. Diets did not affect fasting plasma glucose concentrations, blood glucose response at each glucose time-point after glucose administration, AUCgluc, fasting serum insulin concentrations, area under the insulin curve, and height and appearance time of insulin response. Yet, analysis of acylcarnitines revealed higher propionylcarnitine concentrations (P = 0·03) when fed the prebiotic diet, suggesting colonic fermentation and propionate absorption. Prebiotic supplementation reduced methylmalonylcarnitine (P = 0·072) and aspartate aminotransferase concentrations (P = 0·025), both indicating reduced gluconeogenesis from amino acids. This trial evidenced impaired glucose tolerance and altered insulin response to glucose administration in obese compared to non-obese cats, regardless of dietary intervention; yet modulation of glucose metabolism by enhancing gluconeogenesis from propionate and inhibition of amino acid catabolism can be suggested.

The present study aimed to investigate the effects of two lactation sow feeds, differing in n-6:n-3 ratio, given to sows before parturition on body condition and feed intake, periparturient metabolism (leptin, insulin, triiodothyronine (T3) and thyroxine (T4)), inflammatory parameters (TNFα, IL-6, serum amyloid A (SAA)) and on piglet performance (birth weight, survivability). The feed contained either a low (supplemented with fish oil; f groups) or high (supplemented with sunflower-seed oil; s groups) n-6:n-3 ratio and was administered from 8 d (f8, s8) or 3 d (f3, s3) before parturition until weaning. The level of inclusion of the oil sources was 2 %. Seventy-two sows were randomly allocated 8 d before expected farrowing into four groups: f3, f8, s3, s8. Type of feed had a significant influence on the sows' feed intake during the first 2 d of lactation (s < f), leptin on days 4, 3 and 2 before parturition (f < s), insulin on day 1 after parturition (f < s), T4 on the day before parturition (s < f) and rectal temperature on the day after parturition (f < s). Onset of administration of the feed (3 v. 8 d) had significant effects on leptin on day 2 before parturition (8 < 3), insulin on day 4 before parturition (3 < 8), T3 on day 4 before parturition and on the day after parturition (3 < 8), SAA on day 3 after parturition (8 < 3) and piglet weight during the first days postpartum (3 < 8). In conclusion, under the present conditions, a lactation feed low in n-6:n-3 ratio administered from 8 d before farrowing ensures improved feed intake during the first days postpartum and was associated with a better metabolic change and inflammatory profile in sows in the periparturient period.

The aim of this study was to describe the effects of two diets differing in n-6:n-3 ratio and prepartal feeding regime on gene expression of PPARγ1a/1b, PPARγ1c/1d, PPARγ2, PPARγ coactivator 1A (PPARGC1A), GLUT4, TNFα, adiponectin, leptin, leptin receptor (LEPR), fatty acid binding protein 4 (FABP4), lipoprotein lipase (LPL) in sows' white adipose tissue on the first day of lactation. The relationship between mRNA expression of these genes and circulating insulin, leptin and thyroid hormones was also considered. Diets contained a low (supplemented with fish oil; f group) or a high (supplemented with sunflower oil; s group) n-6:n-3 ratio and were provided from 8 (f8, s8) or 3 d (f3, s3) before parturition (onset day 8 or 3). A low n-6:n-3 ratio reduced the 1 d postpartum expression of PPARγ2 and PPARGC1A but only when applied from 3 d before parturition. Circulating leptin was negatively correlated with mRNA expression of adiponectin, LEPR and LPL, whereas thyroxine was positively correlated with levels of PPARGC1A. In conclusion, the effect of dietary treatments, e.g. altering the n-6:n-3 ratio, around parturition on the expression of crucial genes in nutrient metabolism can be modulated by the duration of application before parturition.

The objective of the present study was to investigate the effects of dietary macronutrient ratio on energy metabolism and on skeletal muscle mRNA expression of avian uncoupling protein (UCP), thought to be implicated in thermogenesis in birds. Broiler chickens from 2 to 6 weeks of age received one of three isoenergetic diets containing different macronutrient ratios (low-lipid (LL) 30 v. 77 g lipid/kg; low-protein (LP) 125 v. 197 g crude protein (N×6·25)/kg; low-carbohydrate (LC) 440 v. 520 g carbohydrate/kg). LP chickens were characterised by significantly lower body weights and food intakes compared with LL and LC chickens (−47 and −38 % respectively) but similar heat production/kg metabolic body weight, as measured by indirect calorimetry, in the three groups. However, heat production/g food ingested was higher in animals receiving the LP diet (+41 %, P<0·05). These chickens also deposited 57 % less energy as protein (P<0·05) and 33 % more as fat. No significant differences in energy and N balances were detected between LL and LC chickens. The diets with the higher fat contents (i.e. the LP and LC diets) induced slightly but significantly higher relative expressions of avian UCP mRNA in gastrocnemius muscle, measured by reverse transcription–polymerase chain reaction, than the LL diet (88 and 90 v. 78 % glyceraldehyde-3-phosphate dehydrogenase respectively, P<0·05). Our present results are consistent with the recent view that UCP homologues could be involved in the regulation of lipid utilisation as fuel substrate and provide evidence that the macronutrient content of the diet regulates energy metabolism and especially protein and fat deposition.

The present study aimed to study the effects of food deprivation and subsequent postprandial changes in plasma somatotrophic and thyrotrophic hormone levels and focused on the inter-relationships between these hormonal axes and representative metabolites of the intermediary metabolism of meal-fed broiler chickens. Male broiler chickens (2 weeks old) were fed a meal of 40–45 g/bird per d for two consecutive weeks (food-restricted (FR) treatment). The daily allowance was consumed in about 30 min. At 4 weeks of age, FR chickens were killed at several time intervals (ten per sampling time) in relation to the daily food allowance: before feeding (about 23·5 h of food deprivation), and at 10, 20, 30 (end of feeding), 40, 50, 60, 90, 120 and 200 min after initiation of feeding. Birds fed ad libitum served as controls (ad-libitum (AL) treatment). Liver tissue was collected for deiodinase type I and type III activity measurements and blood samples for analysis of growth hormone (GH), insulin-like growth factor (IGF)-I, thyroxine (T4), 3,3′,5-triiodothyronine (T3), glucose, non-esterified fatty acids (NEFA), uric acid, triacylglycerol (TG) and lactate levels. Food deprivation caused a shift from lipogenesis to lipolysis and increased fatty acid turnover, a reduction in protein anabolism and reduced metabolic rate. Food intake was followed immediately by a pronounced increase in metabolic rate, initially mainly based on anaerobic mechanisms. Refeeding gradually reversed the fasting-induced alterations in plasma hormone and metabolite levels, but the time course of changes differed between metabolites, which clearly preceded those of the hormones investigated. The order of responsiveness after food provision were glucose>uric acid≥NEFA>lactate>TG for the plasma metabolites and GH>T3>T4>IGF–I for hormones. Based on these different postprandial time courses, several functional relationships are proposed. Glucose is believed to be the primary trigger for the normalisation of the effects of fasting on these plasma variables by restoring hepatic GH receptor capacity, as well as decreasing deiodinase type III activity.

The usefulness of supplementary dietary L-CARNITINE AS AN IMMUNOMODULATOR TO INCREASE ANTIGEN-SPECIFIC ANTIBODY LEVELS WAS ANALYSED IN 2–6-WEEK-OLD BROILERS. THE CHICKENS RECEIVED COMMERCIAL FEEDS EITHER UNSUPPLEMENTED (STARTER FEED 17·8 MG CARNITINE/KG, FINISHER DIET 22·9 MG CARNITINE/KG) OR SUPPLEMENTED WITH l-carnitine (100 mg carnitine/kg added to feed). At 14 d of age, both groups were distributed in equal numbers and sex ratios over two environmentally controlled chambers where temperature (28°) was either reduced immediately to 20°, or gradually to 22° at 36 d of age. Antigen-specific immunoglobulin (Ig)M, IgG, IgA and total Ig responses were measured following two immunizations with bovine serum albumin (BSA). The typical BSA-specific IgM responses followed by IgG responses to the primary immunization were boosted by the secondary immunization. The kinetics of these responses were not altered by l-carnitine treatment. However, BSA-specific total Ig and IgG, but not IgM, responses were significantly increased by dietary l-carnitine supplementation, after both the primary and the secondary immunization. No significant influence of the sex of the chicks or the imposed environmental temperature on Ig responses was found. Temperature treatment and sex, but not l-carnitine supplementation, did significantly influence body-weight gain: cockerels were heavier than females and this became most evident in the second half of the rearing period. Further, lowering the temperature increased body weight. In conclusion, dietary l-carnitine supplementation appeared to be beneficial in enhancing specific humoral responses on vaccination.