The gastric isthmus from D+ and D- broiler lines divergently selected for digestion efficiency shows histological and morphological differences.

Previous results suggested that the gastric function plays a paramount role in digestive efficiency differences between D+ and D- broiler lines divergently selected for AMEn (more feed efficient and less feed efficient, respectively). In this paper we show an histological analysis of the gastric isthmus located between the proventriculus and the gizzard in the D+ and D- lines. Cross-sections were performed using a cryostat (Leica CM30505) and stained with a routine procedure using Mayer's Hematoxylin and Eosin Stain. The surface and shape of the constitutive gastric isthmus tissues were quantified using the image analysis software Image J. The lumen occupied 11% of the whole D- isthmus cross-sectional area against 24% for D+ (P < 0.01). The mucosa relative area (cm(2)/total cross-sectional area) was higher in D- than in D+ [47% (D-) and 39% (D+), P < 0.01]. It was significantly more oval and more folded on the lumen side in D- than in D+ chickens; the muscle layer (muscularis mucosae) of the mucosa was relatively more developed in D- than in D+ (16 and 11% of the section, respectively; P < 0.01). A relationship between these observations and increased gastric motility reported in D- compared with D+ is discussed.

Comparison of gizzard activity between chickens from genetic D+ and D- lines selected for divergent digestion efficiency.

Gizzard motility was compared between chickens from D(+) (high digestion efficiency) and D(-) (low digestion efficiency) genetic lines selected for divergent digestion efficiency, using strain gauge transducers. Motility was recorded continuously during 24 h in 6 birds per line. Two stimuli, a meal distribution after a feed-deprivation period and lighting after a dark period, were tested during the recording period. A functional test with intravenous injection of serotonin performed at the end of the recording day often resulted in a sharp reduction in gizzard motility, without a significant difference between lines. Compared with D(+) birds, gizzard activity in D(-) birds remained high during fasting or dark periods (P < 0.0006), which reduced the effects of stimuli in D(-) birds. So, coordination between bird activity and gizzard motility tended to be reduced in D(-) compared with D(+) birds (P = 0.0018). This coordination was observed to be positively correlated (P = 0.011) with the relative weight (g/kg of BW) of the stomach (gizzard + proventriculus). This experiment pointed out differences in gizzard motility between D(+) and D(-) chicken lines in terms of response to environmental stimuli, characterized by a failure in the gizzard relaxation process in D(-) birds during rest periods.

Sequential feeding using whole wheat and a separate protein-mineral concentrate improved feed efficiency in laying hens.

The effect of feeding nutritionally different diets in sequential or loose-mix systems on the performance of laying hen was investigated from 16 to 46 wk of age. Equal proportions of whole wheat grain and protein-mineral concentrate (balancer diet) were fed either alternatively (sequential) or together (loose-mix) to ISA Brown hens. The control was fed a complete layer diet conventionally. Each treatment was allocated 16 cages and each cage contained 5 birds. Light was provided 16 h daily (0400 to 2000 h). Feed offered was controlled (121 g/bird per d) and distributed twice (4 and 11 h after lights-on). In the sequential treatment, only wheat was fed at first distribution, followed by balancer diet at the second distribution. In loose-mix, the 2 rations were mixed and fed together during the 2 distributions. Leftover feed was always removed before the next distribution. Sequential feeding reduced total feed intake when compared with loose-mix and control. It had lower wheat (-9 g/bird per d) but higher balancer (+1.7 g/bird per d) intakes than loose-mix. Egg production, egg mass, and egg weight were similar among treatments. This led to an improvement in efficiency of feed utilization in sequential compared with loose-mix and control (10 and 5%, respectively). Birds fed sequentially had lower calculated ME (kcal/bird per d) intake than those fed in loose-mix and control. Calculated CP (g/bird per d) intake was reduced in sequential compared with loose-mix and control. Sequentially fed hens were lighter in BW. However, they had heavier gizzard, pancreas, and liver. Similar liver lipid was observed among treatments. Liver glycogen was higher in loose-mix than the 2 other treatments. It was concluded that feeding whole wheat and balancer diet, sequentially or loosely mixed, had no negative effect on performance in laying hens. Thus, the 2 systems are alternative to conventional feeding. The increased efficiency of feed utilization in sequential feeding is an added advantage compared with loose-mix and thus could be employed in situations where it is practicable.

Expression of the glucokinase gene in mule duck liver and glucokinase activities in chicken and mule duck livers.

The presence of glucokinase (GK), a critical enzyme controlling glucose homeostasis, particularly liver glucose utilization in mammals, has long been a matter of debate in avian species because a number of investigators have failed to detect GK activity in the livers of chickens and several other avian species. In this study, we cloned a partial GK cDNA from mule duck livers and measured GK-like activity in the livers of mule ducks and broiler chickens under 2 nutritional states. Liver samples from 5-wk-old meal-fed male broiler chickens (Ross) were obtained from overnight-fasted chickens (BC) and 5 h after an oral saccharose load (6 mL/kg of BW of a 50% saccharose solution) given just before the meal (BS). Liver samples from 15-wk-old mule ducks were collected after an overnight fast (DC) and 12 h after the last overfeeding meal (DO). A partial cDNA ( approximately 600 bp) was obtained from duck livers. It presented 99% identity with chicken partial GK cDNA (gi 44888789) and 82% identity with human GK (gi 15967158). Chicken liver weights represented 1.8 and 3.3% of BW, respectively, for BC and BS (n = 8, P < 0.05). Glucokinase and low-Michaelis constant hexokinase (HK) activity levels were similar in BC (respectively, 0.88 and 1.00 mU/mg of protein). In response to the meal load, GK activity increased significantly (+57%), whereas HK decreased (-46%) in BS. Duck liver weights represented 1.4 and 7.6% of BW, respectively, for DC and DO (n = 8, P < 0.05). In DC livers, GK activity was significantly higher than HK activity (respectively, 1.76 and 0.63 mU/mg of protein). Both activities were significantly increased in DO (2 times, n = 8, P < 0.05). In conclusion, GK is present in ducks as well as chickens, and it is nutritionally regulated in avian species as well as in mammals. Further work will determine whether the higher liver GK activity and GK:HK ratio in DC compared with BC is related to age or BW or linked to the high lipogenic capacity of the duck liver.

Effects of thermal manipulation during early and late embryogenesis on thermotolerance and breast muscle characteristics in broiler chickens.

Genetic selection has significantly improved the muscle development of fast-growing broiler chickens in the last 50 yr. However, improvement in muscle growth has coincided with relatively poor development of visceral systems, resulting in impaired ability to cope with high environmental temperatures. The aim of this study was to elucidate the effects of thermal manipulation (TM) during different periods of embryogenesis on chick hatchability, BW and thermoregulation upon hatching, on their ability to cope with thermal challenge at 42 d of age, and on carcass and breast meat traits. Control embryos were incubated at 37.8 degrees C. The TM embryos were incubated at 37.8 degrees C and treated for 3 h at 39.5 degrees C on the following days of embryogenesis: E8 to E10 [early (EA)], E16 to E18 [late (LA)], and both E8 to E10 and E16 to E18 (EA-LA). Body weight and body temperature (T(b)) were measured at hatching and throughout the growth period as well as during exposure of 42-d-old chickens to a thermal challenge at 35 degrees C for 6 h. The LA and EA chicks exhibited significantly lower T(b) than control chicks (37.9 vs. 38.2 degrees C) at hatching, but during the growth period, differences in T(b) between treated and control chicks decreased with age. Significant hyperthermia (over 44 degrees C) was monitored in all groups during the thermal challenge, but mortality was higher in treated than in control chickens. No effect of treatments on BW was found during the entire growth period. However, breast yield was higher in LA chickens than in controls at slaughter. The EA and EA-LA treatments slightly decreased the ultimate pH of breast meat, whereas the LA treatment had no effect. In conclusion, none of the TM conditions tested in the present study were able to improve long-term thermotolerance in chickens. Late treatment favored breast muscle growth without affecting ultimate pH and drip loss of breast meat.

Insulin immuno-neutralization decreases food intake in chickens without altering hypothalamic transcripts involved in food intake and metabolism.

In mammals, insulin regulates blood glucose levels and plays a key regulatory role in appetite via the hypothalamus. In contrast, chickens are characterized by atypical glucose homeostasis, with relatively high blood glucose levels, reduced glucose sensitivity of pancreatic beta cells, and large resistance to exogenous insulin. The aim of the present study was to investigate in chickens the effects of 5 h fasting and 5 h insulin immuno-neutralization on hypothalamic mRNA levels of 23 genes associated with food intake, energy balance, and glucose metabolism. We observed that insulin immune-neutralization by administration of anti-porcine insulin guinea pig serum (AI) significantly decreased food intake and increased plasma glucose levels in chickens, while 5 h fasting produced a limited and non-significant reduction in plasma glucose. In addition, 5 h fasting increased levels of NPY, TAS1R1, DIO2, LEPR, GLUT1, GLUT3, GLUT8, and GCK mRNA. In contrast, AI had no impact on the levels of any selected mRNA. Therefore, our results demonstrate that in chickens, food intake inhibition or satiety mechanisms induced by insulin immuno-neutralization do not rely on hypothalamic abundance of the 23 transcripts analyzed. The hypothalamic transcripts that were increased in the fasted group are likely components of a mechanism of adaptation to fasting in chickens.

Is there peripheral or ovarian insulin action alteration in broiler breeder hens fed ad libitum?

We investigated whether a change in peripheral glucose homeostasis, a local change in the insulin-related ovarian regulatory system, or both occurred in ad libitum-fed broiler breeder hens compared with feed-restricted counterparts. Feed-restricted (R, from 5 to 16 wk of age) and ad libitum-fed (A) hens from a standard commercial line (S) and an experimental dwarf genotype (E) were studied. Basal and stimulated plasma insulin and glucose concentrations were measured during the prebreeding and laying periods. In the basal state (after 16 h fasting) plasma glucose concentrations were significantly lower in SA chickens (-5% at 17 wk, -7.5% at 32 wk) compared with EA, SR, and ER chickens, with no difference in plasma insulin concentrations (n = 16). In 17-wk-old SA birds, 30 min after oral glucose loading, plasma glucose concentrations increased significantly compared with the basal state and were also significantly lower as compared with SR but did not differ significantly from EA and ER. Plasma insulin concentrations did not differ significantly between genotypes or regimens (n = 16). A potential modification of intracellular mediators involved in the regulation of cell growth and survival in small follicles that were overrecruited in SA compared with SR was also investigated in SA and SR hens at 32 wk. There was no effect of food restriction in phospho-Akt, Akt, phospho-ERK, and phospho-S6 in the small white ovarian follicles (n = 6) in the basal state and after 30 min of refeeding. In conclusion, the present study does not demonstrate any evidence of glucose intolerance during the prebreeding period, specific change in the ovarian small follicle insulin signalling pathway, or both, in laying broiler breeders fed ad libitum compared with feed-restricted hens.

Cyclic variations in incubation conditions induce adaptive responses to later heat exposure in chickens: a review.

Selection programs have enabled broiler chickens to gain muscle mass without similar enlargement of the cardiovascular and respiratory systems that are essential for thermoregulatory efficiency. Meat-type chickens cope with high ambient temperature by reducing feed intake and growth during chronic and moderate heat exposure. In case of acute heat exposure, a dramatic increase in morbidity and mortality can occur. In order to alleviate heat stress in the long term, research has recently focused on early thermal manipulation. Aimed at stimulation of long-term thermotolerance, the thermal manipulation of embryos is a method based on fine tuning of incubation conditions, taking into account the level and duration of increases in temperature and relative humidity during a critical period of embryogenesis. The consequences of thermal manipulation on the performance and meat quality of broiler chickens have been explored to ensure the potential application of this strategy. The physiological basis of the method is the induction of epigenetic and metabolic mechanisms that control body temperature in the long term. Early thermal manipulation can enhance poultry resistance to environmental changes without much effect on growth performance. This review presents the main strategies of early heat exposure and the physiological concepts on which these methods were based. The cellular mechanisms potentially underlying the adaptive response are discussed as well as the potential interest of thermal manipulation of embryos for poultry production.

Further characterization of insulin secretion from the perfused duodenum-pancreas of chicken: a comparison of insulin release in chickens selected for high and low abdominal fat content.

Insulin release from the perfused pancreas was studied in genetically selected fat and lean chickens. The previously described glucose insensitivity of the chicken pancreas cannot be overcome by 5 or 10 mM D-glyceraldehyde, suggesting that the resistance is not related to glucose metabolism before the triose phosphate step. At 42 mM, glucose induced a biphasic insulin release which was specific, since 42 mM mannitol did not elicit insulin release. Arginine (10 mM) or acetylcholine (0.1-1 microM), which in themselves do not cause insulin release, generated a biphasic insulin release in the presence of a low nonstimulating glucose concentration (14 mM); the effect was synergistic. In contrast to the glucose tolerance test observed in vivo, the pancreas from the fat line chicken in response to glucose or glucose plus arginine released significantly less insulin during the first phase. The significance of this defect awaits further elucidation. On the other hand, acetylcholine, a more potent secretagogue, did not reveal any significant difference between fat and lean chickens.

Chicken leptin: properties and actions.

Chicken leptin cDNA shows a high homology to mammalian homologous, with an expression localized in the liver and adipose tissue. It is noteworthy, that the hepatic expression is most likely associated with the primary role that this organ plays in lipogenic activity in avian species. As in mammals, chicken leptin expression is regulated by hormonal and nutritional status. This regulation is tissue-specific and with a high sensitivity in the liver compared to adipose tissue. The blood leptin levels are regulated by the nutritional state with high levels in the fed state compared to the fasted state. The recombinant chicken leptin markedly inhibits food intake as reported in mammals, suggesting the presence of an hypothalamic leptin receptor. The chicken leptin receptor has been identified and all functional motifs are highly conserved compared to mammalian homologous. Chicken leptin receptor is expressed in the hypothalamus but also in other tissues such as pancreas, where leptin inhibits insulin secretion and thus may have a key role in regulating nutrient utilization in this species.

A glucokinase-like enzyme induced in Mule duck livers by overfeeding.

The Mule duck develops a fatty liver in response to overfeeding, which results from a dramatic increase in de novo liver lipogenesis, and thus raises questions regarding the role of glucokinase (GK), a key enzyme regulating carbohydrate metabolism in mammals. However, the presence of GK in avian species is still a matter of debate. The aim of the present study was to characterize a GK-like protein (using an immunological technique) and a GK-like activity (using an enzymatic assay) in duck liver and to measure their respective variations during various stages of overfeeding. Duck liver protein cross-reacted with antibodies directed against mammalian GK yielding a band at 50 kDa, i.e., the same molecular weight as mammalian GK. The intensity of the signal varied significantly between overfed and control ducks but in opposing ways according to the GK antibodies used, which suggests the presence of 2 isoforms of GK in the duck liver as in mammals. Enzymatic analysis demonstrated the presence of glucose phosphorylation activity sensitive to high and low glucose concentrations (high/low ratio between 1.7 and 3.7) in the soluble and particulate fractions of liver homogenates. Glucokinase-like activity per milligram protein was strongly induced by overfeeding, and plasma insulin levels increased concomitantly. More than 80% of total GK-like activity was concentrated in the soluble component from 1 to 13 d of overfeeding. These results suggest that a GK-like enzyme may actively contribute to glucose disposal throughout the overfeeding period in Mule ducks fed a carbohydrate-rich diet.

Thermal manipulation of the embryo modifies the physiology and body composition of broiler chickens reared in floor pens without affecting breast meat processing quality.

Selection in broiler chickens has increased muscle mass without similar development of the cardiovascular and respiratory systems, resulting in limited ability to sustain high ambient temperatures. The aim of this study was to determine the long-lasting effects of heat manipulation of the embryo on the physiology, body temperature (Tb), growth rate and meat processing quality of broiler chickens reared in floor pens. Broiler chicken eggs were incubated in control conditions (37.8°C, 56% relative humidity; RH) or exposed to thermal manipulation (TM; 12 h/d, 39.5°C, 65% RH) from d 7 to 16 of embryogenesis. This study was planned in a pedigree design to identify possible heritable characters for further selection of broiler chickens to improve thermotolerance. Thermal manipulation did not affect hatchability but resulted in lower Tb at hatching and until d 28 post-hatch, with associated changes in plasma thyroid hormone concentrations. At d 34, chickens were exposed to a moderate heat challenge (5 h, 32°C). Greater O2 saturation and reduced CO2 partial pressure were observed (P < 0.05) in the venous blood of TM than in that of control chickens, suggesting long-term respiratory adaptation. At slaughter age, TM chickens were 1.4% lighter and exhibited 8% less relative abdominal fat pad than controls. Breast muscle yield was enhanced by TM, especially in females, but without significant change in breast meat characteristics (pH, color, drip loss). Plasma glucose/insulin balance was affected (P < 0.05) by thermal treatments. The heat challenge increased the heterophil/lymphocyte ratio in controls (P < 0.05) but not in TM birds, possibly reflecting a lower stress status in TM chickens. Interestingly, broiler chickens had moderate heritability estimates for the plasma triiodothyronine/thyroxine concentration ratio at d 28 and comb temperature during the heat challenge on d 34 (h(2) > 0.17). In conclusion, TM of the embryo modified the physiology of broilers in the long term as a possible adaptation for heat tolerance, without affecting breast meat quality. This study highlights the value of 2 new heritable characters involved in thermoregulation for further broiler selection.

Effects of heat exposure on Akt/S6K1 signaling and expression of genes related to protein and energy metabolism in chicken (Gallus gallus) pectoralis major muscle.

In order to improve understanding of the heat-induced changes in muscle growth, we determined the expression of genes related to protein and energy metabolism in the pectoralis major muscle of chickens. We also explored the protein kinase B (PKB also called Akt)/p70 S6 kinase (S6K1)/S6 pathway that mediates anabolic signals thereby regulating metabolism and hypertrophic/atrophic balance. Four-week-old chickens were exposed to 32 or 22 degrees C for 1 week. Chickens from both groups were then fasted for 16 h or left fed, and submitted to an oral administration of glucose-arginine to induce an anabolic response (30-min treatment) or left untreated. High ambient temperature and the associated decrease in feed intake modified the expression of certain energy-related genes (e.g. -40% for PGC-1alpha) and protein metabolism (e.g. about +80% for atrogin-1), but the expression of several muscle metabolism-related genes considered here was unchanged. The capacity for muscle protein synthesis, i.e. RNA/protein ratio, was reduced in warm conditions (approximately -20%). Slightly lower activation of S6 induced by glucose-arginine treatment was found at 32 degrees C compared to 22 degrees C, which might indicate somewhat lower efficiency of mRNA translation. Analysis of glucose/insulin balance suggested changes in glucose metabolism under heat exposure. However, this remains to be characterized.

Do age and feeding levels have comparable effects on fat deposition in breast muscle of mule ducks?

The effects of age (from 1 day post-hatch to 98 days of age) and feeding levels (feed restriction followed by overfeeding v. ad libitum feeding) on lipid deposition in breast muscle (quantity and quality, localisation) of mule ducks were determined in relation to muscle energy metabolism (glycolytic and oxidative), plasma levels of lipids, glucose and insulin, and muscle capacity for lipid uptake (characterised by lipoprotein lipase (LPL) activity). Two periods were defined for age effects on intramuscular lipids in breast muscle: - 1 to 42 days of age when lipids (mainly phospholipids and cholesterol provided by egg yolk) stored in the adipocytes during embryonic life were transferred to the muscle fibres and used for growth and energy requirements, - 42 to 98 days of age when the muscle again stored lipids (mainly triglycerides provided by liver lipogenesis), first in fibres and then in adipocytes.Plasma glucose and insulin levels were not affected by age. Plasma levels of lipids and LPL activity in breast muscle were high at 1 and 14 days of age and then decreased, remaining stable until 98 days of age. Energy metabolism activity in the breast muscle (mainly glycolytic activity) increased with age.Feed restriction, corresponding to 79% of ad libitum intake, applied between 42 and 75 days of age only resulted in decreases in plasma insulin concentration and total lipid content of breast muscle, mainly affecting triglyceride and mono-unsaturated fatty acid (MUFA) levels. Overfeeding increased plasma levels of insulin and lipids while glycaemia remained stable. LPL activity and total lipid levels increased in breast muscle, mainly induced by deposition of triglycerides and MUFA occurring particularly during the 2nd week of this period. Glycolytic energy metabolism decreased.In response to age or feeding levels, muscle lipid levels and composition reflect plasma lipid levels and composition and high muscle lipid levels stimulate oxidative energy metabolism.

Effects of dietary fructose on liver steatosis in overfed mule ducks.

Overfeeding of some waterfowl species results in obesity, which is mainly characterized by a dramatic hepatic steatosis induced by strong accumulation of lipids synthesized from dietary glucose in the liver. In mammals, fructose is known to be able to raise plasma triacylglycerol concentrations significantly; consequently, this may induce obesity. The aim of this study was to assess the effect of partial replacement of dietary glucose provided by corn starch with fructose on metabolism and fatty liver production in the Mule ducks. On the basis of 9.5 kg maize (132,920 kJ) given twice a day for 14 days, a supplementation of 9,800 kJ was provided in form of glucose, sucrose or high fructose corn syrup (HFCS: 50 % glucose, 42 % fructose and 8 % other saccharides). Fatty liver weight in ducks fed with glucose supplementation was 499 +/- 21 g. Sucrose or HFCS supplementation brought about a significant increase in liver weight (+ 18.7 % and + 16.3 % vs. glucose supplementation respectively, p < 0.05). These results suggest that the dietary fructose favors the liver steatosis by increasing hepatic lipogenesis. Postprandial plasma insulin concentrations were similar in ducks fed diets with or without fructose, suggesting that the effect of fructose on liver steatosis is not mediated by insulin.

Specificity of the permissive effect of D-glucose on insulin release in chicken pancreas.

1. As previously shown, 14 mM D-glucose, a non-insulinotropic concentration in isolated chicken pancreas, permits an insulin release in response to D-glyceraldehyde, (D-GA; a glycolytic fuel) and L-leucine or alpha-ketoisocaproic acid (alpha-KIC) (non-glycolytic fuels), which alone are not initiators of insulin release in this species. 2. The "permissive" effect of D-glucose was also observed in the presence of D-mannose (which, as shown herein, is not insulinotropic alone). 3. The specificity of glucose for this "permissive" effect was, therefore, subsequently questioned in the presence of 10 mM alpha-KIC by substituting various glycolytic and non-glycolytic fuels to glucose. 4. D-GA (at 5 and 15 mM), D-mannose (30 and 50 mM), or the association of L-glutamine + L-asparagine permitted an insulin release in response to alpha-KIC. 5. The response was, however, delayed with D-GA, only occasionally with 50 mM D-mannose, and required high concentrations and was delayed in the presence of L-glutamine + L-asparagine as compared to that obtained with 14 mM D-glucose + alpha-KIC. 6. In conclusion, the threshold of fuel-induced insulin release is much higher in the chicken than in mammals and this threshold is most efficiently lowered by glucose.

cAMP and/or acetylcholine permit an insulin response to fuel nutrients in chicken.

1. The possibility that 8-bromo cyclic adenosine monophosphate (8-Br-cAMP) or acetylcholine (ACh) potentiates insulin release in chicken pancreas in response to D-glyceraldehyde (D-GA, a weak insulinotropic fuel), and permits an insulin release in response to D-mannose or alpha-ketoisocaproic acid (alpha-KIC) (two non-insulinotropic fuels in chicken pancreas) is examined. 2. 8-Br-cAMP (1 mM) or ACh (1 microM) permitted a sustained although delayed insulin release in response to D-GA (5 and 15 mM). 3. The resistance to D-mannose (50 mM) or alpha-KIC (10 mM) persisted in the presence of 8-Br-cAMP. 4. At 1 or 100 microM, ACh permitted a slight, immediate and transient insulin output in response to alpha-KIC but not to D-mannose (with one unexplained exception). 5. The simultaneous perfusion of 8-Br-cAMP + ACh increased the basal rate of insulin release, and permitted a large and sustained response to D-mannose. It also greatly increased the immediate response to alpha-KIC + ACh. 6. In conclusion, in chicken pancreas fuel nutrients require the activation of cAMP- and/or ACh-dependent pathways to induce insulin release. Whether this peculiarity is related to the high glycemia of chickens awaits further investigation.