Direct and maternal reduced balanced protein diet influences the liver transcriptome in chickens.

The objective of this study was to evaluate, by means of RNA sequencing, the direct and transgenerational effect of a reduced balanced protein (RP) diet on broiler breeder metabolism. Chickens of the F0 generation were fed a control (C) or RP diet, and their F1 progeny was fed a C or RP diet as well, resulting in four groups of chickens: C/C, C/RP, RP/C and RP/RP. While both direct and maternal effects were seen on body weight, breast muscle weight and abdominal fat weight in the F1 generation, the direct effect was the most dominant one. The liver transcriptome in the F1 generation showed that amino acid metabolism was up-regulated in chickens that received the control feed when compared with their respective contemporaries that received the reduced protein diet. Interestingly, chickens hatched from control-fed hens but reared on the reduced protein diet (C/RP group) activated a fatty acid metabolism, expressing more fatty acid desaturase 1 gene, fatty acid desaturase 2 gene and elongation of very long-chain fatty acids protein 2 gene, when compared with control-fed chickens hatched from control-fed hens (C/C group), while chickens hatched from reduced protein-fed hens that received themselves the same reduced protein diet (RP/RP group) triggered their glucose metabolism more, showing elevated levels of phosphofructokinase gene, 6-phosphofructo-2-kinase/fructose-2,6-biphospatase 4 and fructose-biphosphate aldolase C mRNA compared with the chickens hatched from reduced protein-fed hens but reared on a control diet (RP/C group). This suggests that the maternal protein diet has an impact on the metabolism of broilers when they are reared on a RP diet.

Management factors resulting in a severe reduction in feed intake-induced spiking mortality syndrome in young broiler chicks.

This study aimed to induce spiking mortality syndrome (SMS) in 10-day-old broiler chicks by changing feed particle size (crumble feed to pellet feed) and/or feed source location (from a small feeder at the pen's center to a large feeder at the front of the pen), followed by full day feed deprivation of all broiler chicks on day 11. In total, 396-day-old male Ross 308 broiler chicks were randomly assigned to 4 treatments (Con: without change in feed particle size and feed source location; Par: changing crumble feed to pellet feed on day 10; Loc: changing feed source location on day 10; LocPar: changing both feed particle size and feed source location on day 10). Each treatment consisted of 9 replicate pens with 11 chicks each. Each treatment was applied at 09:00 on days 10 and 11. On both days, chicks with SMS were identified based on clinical symptoms (down in sternal or lateral recumbency, hyperventilation). Plasma glucose, 3, 3', 5-triiodothyronine (T3), thyroxine (T4) concentrations, insulin, and liver glycogen concentrations of chicks without (normal) and with SMS were measured. Proportional organ and digestive tract including content weights were recorded. Broiler behavior was assessed hourly from 08:30 to 17:30 on day 10. On day 10, the Par, Loc, and LocPar groups spent significantly less time feeding and more time lying down compared with the Con group. On days 10 and 11, SMS clinical signs were observed around 2.5 to 3.5 h after the initiation of treatments, and the Loc group had the most SMS morbidity level. Spiking mortality syndrome chicks had significantly less digestive tract contents compared with Normal chicks on day 10. Spiking mortality syndrome was induced successfully with the treatments, according to their significantly reduced plasma glucose, insulin, T3 and T4 concentrations as well as liver glycogen content. A significant correlation between plasma glucose and liver glycogen was observed in SMS chicks. In conclusion, management factors inducing the reduction or absence of feed intake on day 10 or day 11 can trigger the occurrence of SMS in young broiler chicks.

The effect of reduced balanced protein diet on the behavior of female broiler breeders in 2 generations.

The behavior of 2 generations of broiler breeders undergoing a 25% reduced balanced protein (RP) dietary treatment was investigated in the current study. There were 2 treatments for the F0 generation: control (C) breeders fed with standard C diets and RP breeders fed with RP diets. The female progeny of each treatment was again subjected to 2 dietary treatments, resulting in 4 treatments for F1 generation: C/C, C/RP, RP/C, and RP/RP (breeder feed in F0/F1 generation). To maintain the target body weights throughout the trial, breeders on RP diet received on average 10% more feed than C diet breeders. The behavior of the breeders at 8h30 (30 min before feeding at 9h00), 12h00, and 15h30 in weeks 23 and 37 of the F0 generation and in week 6, 11, and 22 of the F1 generation was observed. Litter scratching, feather pecking, and object pecking were occasionally increased by RP diet feeding which indicated feeding frustration. Drinking behavior decreased dramatically by the RP dietary feeding and resulting in a better litter condition which could benefit dust bathing behavior. In addition, feeding the breeders RP diet in the F0 generation decreased litter scratching (week 6) and feather pecking (week 22, 15h30) but increased sitting (week 11, 15h30) and drinking (a tendency in week 6 and a significant effect in week 11) behavior of offspring breeders (F1 generation). In general, breeders fed with reduced balanced protein diets, to some extent, spent less time drinking and their offspring could have an adaptation to the maternal RP diet. The mechanism of this adaptation still needs to be further investigated. In general, positive effects were found by reducing protein level of breeder diets. However, negative side effects such as feeding frustration were also observed, which merit further study.

Dual-energy X-ray absorptiometry is a reliable non-invasive technique for determining whole body composition of chickens.

In this study, a Lunar Prodigy dual-energy X-ray absorptiometry (DEXA) scanner was validated as a technique to estimate chicken body composition in a non-invasive way. Former research has emphasized the importance of validation of every scanner and software version. In a first trial, DEXA estimated body composition for broilers was correlated with chemical carcass analysis to develop prediction equations. As such, those equations can be used in later experiments with chickens to correct DEXA estimations to estimate body composition accurately by DEXA. DEXA estimated fat mass, lean tissue mass, bone mineral content (BMC) and total body mass, which is the sum of fat, lean mass and BMC, were compared to chemically analyzed crude fat, lean mass as the sum of protein and water and body ash content and scale body weight, respectively. Those regression equations were then used in a second trial to determine body composition based upon DEXA for breeders at different ages. In this experiment, fat and lean tissue determined by DEXA, were compared to dissection parameters commonly used for assessing carcass quality, namely breast muscle and abdominal fat. The first trial showed that DEXA provides high correlations for body mass (ρ = 1) and the individual tissue masses separately (ρ ranging between 0.98 and 1). These high correlations allow for accurate prediction of those components with the developed regression equations. Proportional fat and lean tissue were correlated with their chemical counterparts, however, to a lower extent than absolute values due to lower variation between the proportional weights. BMC percentage was not significantly correlated with ash percentage. Furthermore, in trial 2 high correlations were observed between dissection parameters and DEXA-corrected estimations. These correlations show that DEXA can assess carcass quality in breeders without sacrificing the birds. In conclusion, DEXA is a reliable technique to estimate breeder and broiler body composition in a non-invasive way, hence allowing for longitudinal studies over longer periods of time while avoiding sacrificing of birds.

The learning ability and memory retention of broiler breeders: 1 effects of reduced balanced protein diet on reward based learning.

Learning and mental abilities of farm animals are important for their adaptation to new environments and could serve as an indicator for welfare and performance. This study assessed the learning ability and memory retention of broiler breeders through a T-maze test with a reward v. no reward set-up. Feed supplemented with or without meal worms served as a reward. Two trials were conducted with either parent stock (PS) breeders or pure line A (PL) breeders and with the same dietary treatment namely a control (C) group fed with standard commercial diets and a reduced balanced protein (RP) group fed with RP diets (25% reduction of CP and amino acids). To maintain similar target BW, the RP group received on average 10% more feed. A tonic immobility (TI) test was performed to estimate the fearfulness of PS breeders. Most breeders were capable of completing the T-maze tests, but the C group needed less time compared with the RP group when their own feed was given as a reward. However, when meal worms were provided as an extra incentive, the RP group completed the maze significantly faster than the C group. Compared to the C breeders, the RP breeders remained longer in a TI state, indicating a higher level of fearfulness. Long-term memory retention in the T-maze test was observed in both groups. It is concluded that the RP diet had no influences on the learning ability of the reward v. no reward discrimination test and its memory retention of broiler breeders. The increased amount of time the RP breeders needed to solve the test was probably due to a higher sense of fear. Furthermore, the RP diet enhanced the motivation of breeders to obtain an alternative feed such as meal worms.

The learning ability and memory retention of broiler breeders: 2 transgenerational effects of reduced balanced protein diet on reward-based learning.

The effect of reduced balanced protein (RP) diet in the F0 and F1 generation of broiler breeders on the learning ability and memory retention of the F2 generation was investigated by means of a reward v. no reward discrimination T-maze test. There were two treatments for the F0 generation: control (C) group, reared on standard commercial diets, and reduced balanced protein (RP) group, fed with RP diets (25% reduction in CP and amino acids). The female F0-progeny of each treatment was again separated into the two dietary treatments, resulting in four treatments for the F1 generation: C/C, C/RP, RP/C and RP/RP (breeder feed in F0/F1 generation). The RP diets fed breeders received on average 10% more feed than C diets fed breeders to achieve a similar target BW. The F2 generation was composed of four treatments coming from the female F1-progeny of the four treatments and were all fed with C diet (namely C/C/C, C/RP/C, RP/C/C and RP/RP/C). All four F2 generation groups were able to complete the T-maze learning test with a slight difference in success rate but a significant difference within groups was observed regarding the time needed to complete the test. In general, the RP/RP/C group needed more time for completing the test compared with the other three groups and the shortest time was recorded for the RP/C/C group. At similar ages, breeders with early learning experience spent significantly less time in completing the test compared with unexperienced breeders. Long-term memory retention was observed in all four groups whereas the learning ability in solving the test decreased with age. It took longer for the breeders to complete the test at older ages. In conclusion, under our experimental conditions, the RP dietary treatment in previous generations had no influence on the T-maze learning ability and memory retention of broiler breeders of the third generation, although it might have effects on the working performance in the T-maze learning test of F2 generation breeders.

Multigenerational effects of a reduced balanced protein diet during the rearing and laying period of broiler breeders. 2. Zootechnical performance of the F1 broiler offspring.

Several studies in mammals focused on the maternal programming of the metabolism by epigenetic mechanisms, while currently, the consequences of a maternal dietary treatment on the offspring performance of farm animals are of particular interest for commercial purpose. In the present study, we investigated if the zootechnical performance of the progeny was altered by a maternal dietary treatment, being a lower dietary crude protein (CP) of the grandparent and/or parent generation. The multigenerational effects of a reduced maternal CP content were investigated by reducing the dietary CP level by 25% in rearing and laying diets of pure line A breeders. The F0 generation breeders were fed either control (C) or reduced balanced protein (RP) diets. The F1 breeder generation was constructed by dividing the F0 female progeny again over a C or RP diet, resulting in 4 dietary treatments in the F1 generation: C/C, C/RP, RP/C, and RP/RP (letters indicating the diets in, respectively, F0 and F1 generations). The offspring performance was evaluated by a zootechnical and nitrogen retention trial on C and low-protein (LP) broiler diets. For the C broiler diet, the C/RP and RP/RP offspring were characterized by a higher BW from d 35 until d 42 compared to the C/C progeny, whereas the RP/C offspring had an intermediate BW that did not differ from the other groups. A tendency (P = 0.067) towards a better nitrogen retention was observed for the offspring of breeders that received the RP diets in F0 and/or F1 generation compared to the C/C progeny. For the LP broiler diet, the C/RP (P = 0.021) and RP/C (P = 0.001) offspring had a higher BW compared to the C/C progeny during the entire grow-out period. In addition, the C/RP offspring were characterized by a lower FCR from d 28 onwards (P = 0.021). In conclusion, dietary treatments imposed on mother hens can have direct effects on the next generation, as well as indirect effects on multiple generations.

Multigenerational effects of a reduced balanced protein diet during the rearing and laying period of broiler breeders. 1. Performance of the F1 breeder generation.

Studies on mammals and poultry showed that maternal dietary treatments can alter the offspring performance. However, in contrast to rodent studies, little is known about multigenerational dietary manipulations in broiler breeders. The presented research aimed to investigate the effects of a reduction of 25% in the dietary crude protein (CP) level in the F0 generation on the body composition and reproductive performance of F1 broiler breeders. In the F0 generation, breeders were fed either a control (C) or reduced balanced protein (RP) diet, 25% reduction in crude protein and amino acids. Female F0-progeny of each treatment were fed a C or RP diet, resulting in 4 treatments in the F1 breeder generation: C/C, C/RP, RP/C, and RP/RP. The reproductive performance of breeders fed RP diets was negatively influenced by the dietary CP reduction in the F1 generation (P < 0.001). Moreover, breeders descending from hens that received RP diets in the F0 generation showed a significantly reduced reproductive capacity compared to their control fed counterparts (P < 0.001). Breeders fed RP diets in the F1 generation were characterized by higher plasma T3 concentrations (P < 0.001), an increased proportional abdominal fat pad (P < 0.001) and proportional liver weight (P < 0.001). During the rearing phase, the RP fed breeders needed a higher feed allowance, whereas no differences could be observed between the C/C and RP/C or the C/RP and RP/RP breeders. However, breeders originating from birds fed RP diets in the F0 generation needed lower feed allocations in the laying phase to maintain a similar body weight. Egg weight was reduced for the C/RP and RP/RP breeders. At 34 wk of age, eggs from C/RP and RP/RP breeders showed a reduced proportional albumen weight, whereas no effects on egg composition were found at 42 wk of age. It was concluded that prenatal protein undernutrition triggered hens to relocate more energy towards growth and maintenance and less towards reproductive capacity.

The effects of a reduced balanced protein diet on litter moisture, pododermatitis and feather condition of female broiler breeders over three generations.

Protein content reduction in broiler breeder diets has been increasingly investigated. However, broiler breeders reared on low protein diets are characterized by a deterioration of the feather condition. Furthermore, polydipsia induced by controlled feed intake increases litter moisture and as a consequence pododermatitis. This project aimed to study the litter moisture, pododermatitis and feather condition of breeders fed with a 25% reduced balanced protein (RP) diet during the rearing and laying period over three successive generations. The experiment started with two treatments for the F0 generation: control (C) group fed with standard C diets and RP group fed with RP diets. The female F0-progeny of each treatment was divided into the two dietary treatments as well, resulting in four treatments for the F1 generation: C/C, C/RP, RP/C and RP/RP (breeder feed in F0/F1 generation). The RP diet fed breeders received on average 10% more feed than C diet fed breeders to achieve the same target BW. The female F1-progeny of each treatment were all fed with C diets which resulted in four treatments for the F2 generation: C/C/C, C/RP/C, RP/C/C and RP/RP/C (breeder feed in F0/F1/F2 generation). Litter moisture, footpad and hock dermatitis were recorded at regular intervals throughout the experimental period in all three generations. For the F0 and F1 generation, the pens of breeders receiving C diets had significantly higher litter moisture than the RP diets fed groups (P<0.05), resulting in an elevated footpad dermatitis occurrence (FDO) (P<0.05). No difference was found in the F2 generation. The feather condition was scored during the laying period for each generation. F0 and F1 breeders reared on the RP diets had poorer feather condition than those receiving the C diets (P<0.05). The C/RP breeders had a significantly poorer feather condition than RP/RP breeders (P<0.05). For the F2 generation, RP/RP/C breeders had a significantly better feather condition compared with the other three groups (P<0.05). The RP/C/C breeders were significantly better feathered than C/C/C breeders (P<0.05). In conclusion, providing RP diets to broiler breeders improved litter condition and hence reduced FDO whereas impaired feather condition. Furthermore, positive transgenerational effects of the maternal RP diets on the feather condition may be inferred, hence potentially altering the welfare status.

Feeding broiler breeders a reduced balanced protein diet during the rearing and laying period impairs reproductive performance but enhances broiler offspring performance.

Mammalian studies have shown that nutritional constraints during the perinatal period are able to program the progeny (metabolism, performance). The presented research aimed to investigate if broiler breeders and their offspring performance could be influenced by reducing the dietary crude protein (CP) level with 25%. A total of 160 day-old pure line A breeder females were randomly divided over 2 dietary treatments. The control group was fed commercial diets, whereas the reduced balanced protein (RP) breeders received an isoenergetic diet that was decreased with 25% in dietary CP and amino acid during their entire lifespan. The RP birds required an increased feed allowance, varying between 3 and 15%, to meet the same BW goals as their control fed counterparts. The difference in feed allocations and reduction of the dietary CP level resulted in a net protein reduction varying between 14 and 23%. At wk 27 and 40, the body composition of the breeders was changed as a result of the dietary treatment. At both ages, the proportional abdominal fat pad weight of the RP breeders was increased (P < 0.001), whereas the proportional breast muscle weight was only higher at wk 27 in the control group compared to the RP group (P < 0.001). Egg weight (P < 0.001) and egg production (P < 0.001) was decreased for the RP fed birds. The lower dietary CP level reduced the proportional albumen weight of the RP eggs (P = 0.006). Male offspring from RP breeders were characterized by an increase in BW from 28 d until 35 d of age (P = 0.015). Moreover, female progeny of RP breeders showed a reduced FCR (P = 0.025), whereas male progeny showed a tendency (P = 0.052) towards a lower FCR at 5 wk of age. In conclusion, lowering dietary CP levels in rearing and laying phase of breeders had a negative effect on breeder performance but enhanced live performance of the offspring.

Prenatal tolbutamide treatment alters plasma glucose and insulin concentrations and negatively affects the postnatal performance of chickens.

To examine the relationship of insulin and glucose, broiler embryos were subjected to acute or prolonged hypoglycemia during the late embryonic phase by, respectively, injecting once (at embryonic day [ED] 16 or 17) or on 3 consecutive days (ED 16, 17, and 18) with tolbutamide (80 µg/g embryo weight), a substance that stimulates insulin secretion from the pancreas. After 1 tolbutamide injection, a prolonged (32 h) decrease of plasma glucose and a profound acute increase in plasma insulin were observed. The 3 consecutive tolbutamide injections induced hypoglycemia for 4 days (from ED 16 to ED 19). The postnatal performance after 3 consecutive tolbutamide injections in broiler embryos was also investigated. Body weight was lower in tolbutamide-treated chickens from hatch to 42 d compared with sham (P = 0.001) and control (P < 0.001) chickens. Feed intake was lower in the tolbutamide group from hatch to 42 d as compared with sham (P = 0.007) and control (P = 0.017) animals. In addition, at 42 d, plasma glucose concentrations, after an insulin injection challenge (50 µg/kg body weight), were higher in tolbutamide-treated chickens compared with the sham and the control group as were their basal glucose levels (P value of group effect <0.001). In conclusion, tolbutamide treatment during the late embryonic development in broilers resulted in prolonged hypoglycemia in this period and negatively influenced the posthatch performance.

Partial albumen removal early during embryonic development of layer-type chickens has negative consequences on laying performance in adult life.

To examine the importance of albumen as a protein source during embryonic development on the posthatch performance of laying hens, 3 mL of the albumen was removed. At hatch, no difference in BW could be observed. Chicks from the albumen-deprived group had a lower residual yolk weight due to higher yolk utilization. During the rearing phase (hatch to 17 wk of age), the BW of the albumen-deprived pullets was lower compared with the control and sham pullets. The feed intake of the albumen-deprived pullets was also lower than the control pullets. However, during the laying phase (18 to 55 wk of age) these hens exceeded the control and sham hens in BW, although this was not accompanied by a higher feed intake. The albumen-deprived hens exhibited a lower egg production capacity as demonstrated by the reduced egg weight, laying rate, and egg mass and increased number of second grade eggs. In addition, the eggs laid by the albumen-deprived hens had a higher proportional yolk and lower proportional albumen weight. In conclusion, prenatal protein deprivation by albumen removal caused a long-lasting programming effect, possibly by differences in energy allocation, in favor of growth and maintenance and impairing reproductive performance.