In ovo L-arginine supplementation stimulates myoblast differentiation but negatively affects muscle development of broiler chicken after hatching.

In this study, we tested the hypothesis that in ovo feeding (IOF) of L-arginine (L-Arg) enhances nitric oxide (NO) production, stimulates the process of myogenesis, and regulates post-hatching muscle growth. Different doses of L-Arg were injected into the amnion of chicken embryos at embryonic day (ED) 16. After hatching, the body weight of individual male chickens was recorded weekly for 3 weeks. During in vitro experiments, myoblasts of the pectoralis major (PM) were extracted at ED16 and were incubated in medium containing 0.01 mm L-Arg, 0.05 mm L-Arg, and (or) 0.05 mm L-nitro-arginine-methyl-ester (L-NAME), an inhibitor of nitric oxide synthase (NOS). When 25 mg/kg L-Arg/initial egg weight was injected, no difference was observed in body weight at hatch, but a significant decrease was found during the following 3 weeks compared to that of the non-injected and saline-injected control, and this also affected the growth of muscle mass. L-NAME inhibited gene expression of myogenic differentiation antigen (MyoD), myogenin, NOS, and follistatin, decreased the cell viability, and increased myostatin (MSTN) gene expression. 0.05 mm L-Arg stimulated myogenin gene expression but also depressed muscle cell viability. L-NAME blocked the effect of 0.05 mm L-Arg on myogenin mRNA levels when co-incubated with 0.05 mm L-Arg. L-Arg treatments had no significant influence on NOS mRNA gene expression, but had inhibiting effect on follistatin gene expression, while L-NAME treatments had effects on both. These results suggested that L-Arg stimulated myoblast differentiation, but the limited number of myoblasts would form less myotubes and then less myofibers, while the latter limited the growth of muscle mass.

Effects of dietary supplementation of methionine and its hydroxy analog DL-2-hydroxy-4-methylthiobutanoic acid on growth performance, plasma hormone levels, and the redox status of broiler chickens exposed to high temperatures.

Heat stress is known to impair performance and to induce oxidative stress in poultry. The aim of the present study was to compare the effects of dietary supplementation of dl-methionine (dl-M) or the synthetic analog 2-hydroxy-4-methylthiobutanoic acid (dl-HMTBA) on broiler growth performance, plasma hormone levels, and some oxidative stress-related parameters under conditions of chronic exposure to high temperatures (HT). From 2 to 6 wk of age, male broiler chickens were reared under either a constant temperature of 32°C until 6 wk of age or a normal temperature scheme (gradual decrease to 18°C at 5 wk of age). Chicks in both the normal and HT treatments were provided with a commercial grower diet supplemented with either 1.0 or 1.2 g/kg of dl-M or 1.0 or 1.2 g/kg of dl-HMTBA. Because there were no effects of supplement dose, data were pooled over both doses within each temperature treatment. The chronic HT treatment impaired feed intake and BW gain, but these negative effects were less pronounced when the chickens received dl-HMTBA. Exposure to HT was also associated with decreased (P < 0.001) plasma thyroid hormones and increased (P < 0.0001) plasma corticosterone levels. At 4 wk of age, and irrespective of the supplemental source, chickens subjected to HT were characterized by significantly lower plasma TBA-reactive substance levels. In contrast, at 6 wk of age, plasma TBA-reactive substance levels were significantly increased by HT, but this effect was observed only for the chickens receiving dl-M and not for those receiving dl-HMTBA. High temperatures induced a significant increase in hepatic total glutathione (GSH) and oxidized GSH levels, regardless of the supplemental source. However, the hepatic ratios of reduced GSH to total GSH and reduced GSH to oxidized GSH were highest in chickens supplemented with dl-HMTBA. In conclusion, dl-HMTBA supplementation partially prevented the growth-depressing effects of chronic heat exposure compared with dl-M supplementation. It can be inferred that dl-HMTBA is more efficient in alleviating HT-induced oxidative damage because of a more favorable reduced GSH-to-total GSH ratio.

Evaluation of supplementary stevia (Stevia rebaudiana, bertoni) leaves and stevioside in broiler diets: effects on feed intake, nutrient metabolism, blood parameters and growth performance.

A perennial schrub, stevia, and its extracts are used as a natural sweetener and have been shown to possess antimicrobial properties. Stevia contains high levels of sweetening glycosides including stevioside which is thought to possess antimicrobial and antifungal properties. Little is known about the nutritional value of the schrub in livestock. This study determined the potential use of the shrub as a prebiotic animal feed supplement in light of the recent ban on the use of antibiotics in animal feed and the role of its constituent stevioside in the effects of the shrub. Male Cobb broiler chicks were fed a basal broiler diet without antibiotic but with performance enhancing enzyme mix (positive control), a basal diet without antibiotic and enzymes (negative control), or diets in which 2% of the negative control diet was replaced with either dried ground stevia leaves or 130 ppm pure stevioside during 2 week starter and 2 week grower periods. Body weight gains, feed conversion, abdominal fat deposition, plasma hormone and metabolites and caecal short chain fatty acids (SCFA) were measured in the broilers at 2 and 4 weeks of age. There was no significant effect of the treatments on feed intake during the starter period but birds fed diet supplemented with stevia leaves and stevioside consumed more feed (p < 0.05) than those fed the positive control diet during the grower period. Weight gain by birds fed the positive control and stevioside diets was higher (p < 0.05) than those fed other diets only during the starter period. Feed/gain ratio of birds fed the positive control and stevioside diets was superior (p < 0.05) to others. There was no effect of the treatments on nutrient retention and water content of the excreta. Dietary stevia leave and stevioside decreased total concentration of SCFA and changed their profile in the ceca. There was no effect of the treatments on pancreas weight. Dietary stevia reduced blood levels of glucose, triglycerides and triiodothyronine (T(3)) but had no effect on non-esterified fatty acids. In contrast, stevioside only decreased T(3). Both the stevia leaves and stevioside diets significantly increased abdominal fat content. It is concluded that dietary enzyme growth promoters are beneficial to the broilers only during the starter stage and that inclusion of stevia leaves or stevioside has no beneficial effect on the performance of broilers.

Impaired development of broiler chickens by stress mimicked by corticosterone exposure.

The effects of corticosterone (CORT) administration on the development of muscular tissues of broiler chickens (Gallus gallus domesticus) fed with diets differing in lipid content were investigated. The experimental chickens were given one of two experimental diets: high lipid diet (9.9% crude fat) or control diet, from 21 d of age. At 28 d of age, half of the chickens in each dietary treatment were exposed to CORT treatment, supplemented with 30 mg CORT/kg diet for 12 days, while the other half continued to consume the former diet. The zootechnical parameters were recorded at 21, 28, 35 and 39 d, and a blood sample was obtained from 8 birds of each group, respectively. The growth performance of broiler chickens was significantly depressed by CORT administration, but not by dietary treatment. Corticosterone treatment resulted in enhanced energy expenditure. The results indicate that the development of breast muscle was more susceptible to stress mimicked by CORT administration. The results suggest that corticosterone administration enhanced hepatic fatty acid synthesis and resulted in the redistribution of energy to abdominal store from peripheral tissues. Diet rich in lipid content was favorable to the central fat deposit in stressed broiler chickens.

Oxidative stress induced by corticosterone administration in broiler chickens (Gallus gallus domesticus) 1. Chronic exposure.

The effects of long-term dietary administration of corticosterone (CORT) on the induction of oxidative injury in broiler chickens (Gallus gallus domesticus) were evaluated. The experimental broiler chickens were fed with a diet supplemented with 30 mg CORT/kg diet for 2 weeks from 14 days of age onwards, while control chickens continued to consume the control diet. The growth performance parameters were recorded weekly, and a blood sample was obtained from eight birds of both groups before CORT administration and at 3, 7 and 14 days after treatment. The results showed that chronic CORT administration resulted in enhanced proteolysis and gluconeogenesis. Furthermore, CORT administration may initially induce the formation of reactive oxygen species (ROS) as indirectly reflected by an increase in lipid peroxidation. However, the significantly increased plasma uric acid (UA) and ceruloplasmin (CP) levels after 3 days of treatment indicates an enhancement of the nonenzymatic antioxidant capacity during stress, and in this way, the development of a more severe oxidative injury is alleviated. Broiler chickens seem to adapt to high circulating CORT levels in terms of their redox homeostasis after 3 days of treatment under the present experimental conditions.

Free diet selection by broilers as influenced by dietary macronutrient ratio and corticosterone supplementation. 1. Diet selection, organ weights, and plasma metabolites.

Male broiler chickens (aged 21 d) were allowed to chose freely for 14 d between three diets in which only one specific macronutrient (protein, lipid, or carbohydrate) was isocalorically substituted for one other macronutrient, but otherwise (nearly) isocaloric and composed of the same ingredients. The three diets were low protein (LowCP; 15.81% CP; 6.56% lipid; 50.78% carbohydrate), low lipid (LowL; 19.63% CP; 3.01% lipid; 51.12% carbohydrate), and low carbohydrate (LowCHO; 19.50% CP; 7.72% lipid; 44.00% carbohydrate). The chickens either received 0, 30, or 45 mg of corticosterone (CORT) per kg diet. As a percentage of their total intake, unsupplemented chickens consumed 24.0, 71.4, and 4.6% of the LowCP, LowL, and LowCHO diets, respectively, giving a total CP, L, and CHO intake of 282, 61, and 765 g, respectively. The addition of CORT significantly changed the diet selection, as compared to the unsupplemented chickens, CORT chickens consumed a greater percentage from the LowCP (35%), less from the LowL (55%), and again more from the Low-CHO (10%) diet. On the other hand, total feed consumption, macronutrient, and ME intake were not altered significantly by CORT supplementation, probably because of the close similarity of the diets. Corticosterone-supplemented chickens manifested hyperglycemia, hyperlipidemia, and uric acidemia suggesting insulin resistance, increased lipogenesis and protein catabolism, respectively. The elevated plasma creatine kinase (CK) activities of CORT chickens are also suggestive for decreased muscle cell membrane stability. Furthermore, CORT chickens were characterized by increased proportional weights of liver, abdominal fat pad, proventriculus, and gizzard, whereas an involution of spleen and bursa was observed. In conclusion, the present results suggest that high circulating levels of CORT as in the case of stress results in metabolic alterations, which in turn, affects diet preference as a compensatory mechanism to adapt energy and nutrient metabolism.

The effects of dietary L-carnitine supplementation on the performance, organ weights and circulating hormone and metabolite concentrations of broiler chickens reared under a normal or low temperature schedule.

1. The present study examined the effects of the addition of 100 mg/kg L-carnitine to the basal starter (containing 17.8 mg/kg L-carnitine) and finisher (containing 22.9 mg/kg L-carnitine) diets on performance, organ weights and plasma hormone and metabolite concentrations of male and female broiler chickens. The broiler chickens were reared either in a room with a normal temperature (NT) program or with a low temperature (LT) program (rapid decrease from 28 degrees C to 20 degrees C at 14 d of age). 2. Broiler chickens reared under the LT schedule consumed more food and attained a greater body weight at 42 d of age than their counterparts reared under NT schedule, without any difference in food efficiency or abdominal fat content. Dietary L-carnitine supplementation had no significant effects on any of these production parameters, except for a reduction in the abdominal fat content of female NT chickens. However, the LT schedule and dietary L-carnitine supplementation greatly increased absolute and proportional heart weights. The elevated heart weights were not due to right ventricle hypertrophy. 3. Both the LT program and L-carnitine supplementation increased circulating plasma triiodothyronine concentrations. There were also some transient effects of both experimental variables on plasma growth hormone, glucose and triglyceride concentrations. 4. L-carnitine did not improve broiler performance. However, this result does not mean that L-carnitine supplementation cannot have beneficial effects in other circumstances. In view of the elevated proportional heart weights, it can also be argued that L-carnitine is a potential agent for reducing the incidence of metabolic diseases in broiler chickens.

Teratological studies in defatted jojoba meal-supplemented rats.

To look for possible developmental effects in the offspring of jojoba meal-treated Wistar rats, and to distinguish between the effects of reduced food intake and the specific developmental effects of jojoba meal itself, mated female rats were divided into three groups of 20 rats. They received during gestation: (a) normal rodent food (control group); (b) normal rodent food supplemented with 3% defatted jojoba meal (jojoba group); or (c) normal rodent food pair-fed with the jojoba group (pair-fed group). The jojoba meal group showed approximately 30% inhibition of food intake. Ten rats from each group were killed on gestation day 21. Compared to the control group, foetal body weight was reduced in both the jojoba and pair-fed groups, with a greater reduction in the jojoba group. Skeletal ossification was retarded to the same extent in both the jojoba and pair-fed groups. The other 10 rats from each group were left to produce litters. Compared with controls, the body weight of the pups was lower in both the jojoba and pair-fed groups; the reduction was slightly greater in the jojoba group, but this difference disappeared after 1 week. The offspring showed no other abnormalities and reproduced normally. We conclude that, at the dose used, the retardation in foetal skeletal ossification, induced by jojoba meal supplementation during gestation, is due to food intake inhibition. Moreover, the lower birth weight of the young of jojoba-treated dams compared with the pair-fed group is merely due to a lower body weight gain during gestation.

Evaluation of jojoba meal as a potential supplement in the diet of broiler breeder females during laying.

1. This study was undertaken to investigate whether jojoba meal can be used as a food supplement during the laying period of chickens. 2. The size of eggs laid were smaller and the overall production rate was lower compared to control birds on food without jojoba meal supplementation. Furthermore, both ovary and oviduct weights were lower in jojoba fed birds. 3. This lowering of egg size and production rate was caused by factors present in jojoba which interfere with follicle growth, yolk deposition, progesterone production and the follicular maturation processes, resulting in the ovulation of smaller follicles and a lower ovulation rate.

The reduction of heat production in exercising pigeons after L-carnitine supplementation.

Four groups (CS,CR,PS,PR) of nine trained male racing pigeons were deprived of feed for 1 d and then subjected to a respiration chamber test in order to study the effect of oral 1-carnitine supplementation on the energy metabolism during flight. One week before, groups CS and CR were orally supplemented with 90 mg of 1-carnitine daily, whereas PS and PR were given a placebo. Groups CS and PS underwent flight simulation by electrostimulation of the breast muscles. Flight simulation increased heat production, kept respiratory quotient from decreasing, decreased thyroxine levels, and increased weight loss. L-Carnitine decreased the rise in heat production during electrostimulation but did not influence respiratory quotient, weight loss, or thyroid hormones. L-Carnitine supplementation in pigeons improves fatty acid combustion efficiency during heavy exercise.

Reproductive performance of rats treated with defatted jojoba meal or simmondsin before or during gestation.

The effects on food intake, growth and reproductive performance parameters of defatted jojoba meal and pure simmondsin, an extract from jojoba meal, were compared in female Wistar rats. Rats fed 0.15% simmondsin or 3% defatted jojoba meal (equivalent to 0.15% simmondsin) for 8 weeks before conception showed a similar reduction in food intake (about 20%) and a similar growth retardation compared with controls. Both treatments induced a reduction in the number of corpora lutea on gestation day 16: this effect could be ascribed to the lower food intake before conception because it was also observed in rats pair-fed to the treated ones. Rats given feed containing 0.15% simmondsin or 3% defatted jojoba meal during days 1-16 of gestation showed a similar reduction in food intake relative to controls. Foetal and placental weights were reduced, relative to controls, to a similar extent in both groups, and the reductions were slightly greater than in the corresponding pair-fed groups. We conclude that the effects on food intake, growth and reproductive performance that were seen after feeding rats defatted jojoba meal were due to the simmondsin content of the meal. The simmondsin induced reduction in food intake and probably also a relative protein shortage.

The prophylactic effect of vitamin C supplementation on broiler ascites incidence and plasma thyroid hormone concentration.

In a two-factorial experiment, 420 1-day-old male commercial broiler chickens were randomly divided and fed a basal diet with or without vitamin C (500 parts/10(6)). Half of the birds within each group receiving the same kind of feed were fed a T(3 )supplemented diet from day 1, in order to increase the ascites incidence. Weekly body weight and feed intake were measured. Venous blood samples were taken from 10 birds per group weekly from day 14 on, for haematocrit measurements, blood gas analysis, measurement of thyroid hormones and lactate/pyruvate ratios. Ascites mortality was recorded daily. Dietary T(3) significantly increased ascites mortality. Adding vitamin C to the feed significantly reduced ascites mortality while it had no effect on performance parameters. Plasma thyroid hormone levels were significantly reduced by dietary vitamin C administration, but neither haematocrit nor blood gas levels were influenced.

Devazepide reverses the anorexic effect of simmondsin in the rat.

Simmondsin, a glycoside extracted from jojoba meal (Simmondsia chinensis), causes a reduction in food intake after oral administration. To investigate the mechanism by which simmondsin reduces food intake, fasted and free-feeding rats were given simmondsin-supplemented food and simultaneously injected with devazepide, a specific antagonist of peripheral-type cholecystokinin receptors (CCKA receptors). In free-feeding rats, supplementation of food with 0.5% simmondsin caused a reduction in food intake of +/- 40% in the period of 4 h following food presentation. Intraperitoneal injection of 100 micrograms devazepide/kg body weight prevented this effect. In rats fasted for 20 h, the food intake in the 30 min after presentation of food supplemented with 0.15% or 0.50% simmondsin was reduced in a dose-related manner; this was also inhibited by simultaneous application of devazepide. It is suggested that peripheral CCKA receptors are involved in the effect of simmondsin on food intake. However, a direct effect of simmondsin on CCKA receptors has been excluded, since simmondsin was unable to cause contraction of the guinea-pig gallbladder in vitro.

Jojoba meal (Simmondsia chinensis) in the diet of broiler breeder pullets: physiological and endocrinological effects.

The present studies evaluated the ability of jojoba meal (JO) to inhibit feed intake of broiler breeder pullets to limit body weight gain as recommended by the breeder company. A first experiment, using graded levels of JO supplementation (0 to 12%), was conducted to establish appropriate JO supplementation. Adequate reduction of growth rate was obtained with 4% JO supplementation. However, notwithstanding their similar growth rate, 4% JO chickens consumed considerably more feed compared with feed-restricted chickens. The dose-dependent impairment of feed intake with increasing levels of JO supplementation was also associated with increased plasma growth hormone and thyroxine and with decreased plasma insulin-like growth factor-I and triiodothyronine concentrations compared with 0% JO chickens. A second experiment included a pair-fed group. Notwithstanding their similar feed intake, 4% JO chickens gained significantly less body weight compared with their pair-fed counterparts. The 4% JO chickens also had a longer feed transit time per kilogram body weight. Again, circulating levels of the somatotrophic and thyrotrophic hormones were altered according to the dietary treatment. From all these observations, it was concluded that the growth retardation caused by JO supplementation was provoked by an inhibition of appetite linked with the simmondsin content of JO as well as by other antinutritional compounds affecting digestibility.

Accumulation of zinc in egg yolk, ovarian follicles and organs after forced resting by high dietary zinc.

1. Eighteen Warren SSL hens of 71 weeks of age were forced-moulted by ad libitum feeding of a high-zinc diet (10,000 ppm zinc for 2 days followed by 5,000 ppm zinc-supplement diet for 4 days). From the start of the treatment, eggs were collected and 3 hens were slaughtered on days 0, 2, 3, 4, 5 and 6 of the study. 2. Zinc analyses were carried out on the different components of the eggs and on liver, pancreas, kidney, different yolky follicles of the ovary and various segments of the oviduct. 3. Seven-, six- and threefold increases in zinc concentration were found in pancreas, liver and kidney, respectively. 4. The shell gland and isthmus, but not the magnum, also showed slight but significant increases in Zn content. 5. Zinc accumulation was also high and almost identical in ovarian follicles F1 to F4 but slightly less in F5 and F6 follicles. 6. In the egg, a significant increase in zinc concentration was only observed in the yolk.

Changes in progesterone metabolism in the chicken hypothalamus during induced egg laying stop and molting.

In the present study, we have established and validated a radioenzyme assay which permits us to quantify progesterone metabolism in the chicken brain. Progesterone metabolism was then studied in five brain areas obtained by microdissection from the telencephalon (part of the lobus paraolfactorius immediately rostral to the preoptic area), the preoptic area, and the hypothalamus. Three metabolites of progesterone were produced in large amounts in these brain regions and were quantified in this study: 5 beta-pregnane-3,20-dione (5 beta-DHP) as well as its metabolite 3 alpha-hydroxy-5 beta-pregnane-20-one (5 beta,3 alpha-ol) and 5 alpha-pregnane-3,20-dione (5 alpha-DHP). The unmetabolized progesterone was also recovered and quantified. The 5 beta-reduction of progesterone (production of 5 beta-DHP and 5 beta,3 alpha-ol) was very active but its 5 alpha-reduction (production of 5 alpha-DHP) was almost absent in the lobus paraolfactorius. An opposite pattern of metabolism was found in the preoptic area and the hypothalamus (higher 5 alpha- but lower 5 beta-reductase activity). The changes in progesterone metabolism in these brain areas were then studied in groups of hens submitted to induced egg laying stop and molting. A significant decrease in progesterone 5 alpha-reduction was found in the median hypothalamus of hens during the period of molt. Simultaneously, the experimental procedures induced significant decreases in the production of 5 beta-DHP by the lobus paraolfactorius, anterior, and medial hypothalamus but induced a significant increase in the production of this metabolite in the preoptic area. These changes are likely to be involved in the control of reproductive functions including sexual behavior and secretion of luteinizing hormone-releasing hormone, and a number of possible causal mechanisms are presented. These should now be tested experimentally especially in view of the very limited information which is now available on the biological effects of the metabolites of progesterone.

The presence of vasotocin and mesotocin in serum and the hypothalamo-neurohypophyseal axis of chick embryos before hatching and in posthatch chicks.

Decreasing hematocrit values were observed in chick embryos, from Day 17 to 1 day posthatching. Arginine vasotocin (AVT) and mesotocin (MT) were determined radioimmunologically in serum, neural lobe, and hypothalamic tissue. Serum levels of AVT and MT were significantly increased on Day 18 of incubation compared to Day 17. Thereafter AVT levels decreased, reaching at hatching levels even below Day 17. Serum MT remained elevated on Day 19, but decreased thereafter to the concentration values of Day 17. The amount of AVT in the preoptic hypothalamus and infundibulum was maximal on Day 19, whereas in the neural lobe the maximum was attained on Day 20. The MT content of the infundibulum and neural lobe reached a maximum plateau on Day 18. These results suggest that a stimulation of the hypothalamo-neurohypophyseal axis in the chick embryo occurs at the end of incubation. A possible causal relation with the observed decreasing hematocrit values is discussed.