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In ovo feeding, injecting nutrients into the amnion of the avian embryo, may enhance jejunal nutrient uptake, activity of the intestinal enzymes, and posthatch growth. This hypothesis was tested in the following in ovo feeding (IOF) experiments. In experiment 1, 400 eggs were evenly distributed among 4 nutritional treatments at 23 d of embryonic development (23E) and administered 1 of 4 treatments as a 2 x 2 factorial arrangement of arginine (ARG 0, 0.7%) and beta-hydroxy-beta-methyl-butyrate (HMB 0, 0.1%). Tissues were assayed for maltase, sucrase, and leucine aminopeptidase (LAP) at 25E, hatch, and 3, 7, and 14 d. In experiment 2, all IOF procedures were repeated and treatments were administered at 21E: injected or noninjected control, 21% egg white protein (EWP), 21% EWP + 0.1% HMB. In experiment 3, two hundred eggs were evenly distributed among the following treatments at 23E: noninjected control or 0.7% ARG + 0.1% HMB + 21% EWP. Jejunal samples were assayed for glucose or alanine uptake at 23E, 25E, and hatch (experiment 2), and hatch and 7 d (experiment 3), respectively. All poults were fed a turkey starter diet ad libitum immediately upon hatching. There was a highly significant HMB x ARG interaction on jejunal sucrase, maltase, and LAP activities at 25E and 14 d. Poults in ovo (IO) fed HMB + ARG had approximately a 2- to 3-fold increase in jejunal sucrase, maltase, and LAP activities at 25E, and a 3-fold increase at 14 d, over other treatments. Poults IO fed EWP + HMB (experiment 2) had enhanced glucose uptake at 25E, whereas poults IO fed ARG + HMB + EWP (experiment 3) had enhanced alanine uptake at hatch and 7 d. These studies demonstrate that IOF ARG, HMB, and EWP may enhance jejunal nutrient uptake and digestion in turkeys.

In ovo feeding (IOF), injecting dietary components into the amnion about 1 d prior to internal pipping, may enhance growth by altering glycogen status. This hypothesis was evaluated with 5 IOF solutions containing protein, beta-hydroxy-beta-methylbutyrate (HMB), and carbohydrate. Four IOF treatments were arranged as a factorial of 2 levels of egg white protein (EWP; 0 and 18%) and 2 levels of HMB (0 and 0.1%). An IOF solution of carbohydrates (S; 20% dextrin and 3% maltose) was evaluated for contrast purposes. At 23 d of incubation, 1.5 mL of IOF solution was injected into the amnion of 100 eggs per treatment. At hatch, feed and water were provided ad libitum. At hatch and 3 and 7 d of age, BW were determined, and 10 poults per treatment were sampled to determine liver (LG) and pectoralis muscle (PC) glycogen content. Poults on IOF treatments A (18% EWP), B (18% EWP + HMB), and D (HMB) weighed 6.0, 2.7, and 3.3% more than the controls at hatch, respectively (P < 0.05) with an EWP x HMB interaction (P < 0.05) sustained to 3 and 7 d only in treatment D (P < 0.005). At hatch, A and D poults had greater percentages of PC (P < 0.05) than controls, and the percentage of PC in treatment D was sustained until 7 d. Total LG was enhanced by A and B at 7 d (P < 0.05) over the controls, whereas total PC glycogen was enhanced at 7 d by IOF treatment D (P < 0.05). The IOF A and S poults had greater BW than the controls at hatch only (P < 0.05). The IOF treatment A had greater LG at hatch (P < 0.05), but by 7 d, A and S had greater LG than controls (P < 0.05). Poults fed S in ovo had enhanced total PC glycogen over controls, whereas poults on treatment A had less total PC glycogen than controls (P < 0.05). The results of this experiment demonstrate that IOF of A or S poults may enhance hatch BW and glycogen status of poults during the neonatal period by inclusion of HMB.

Early posthatch satellite cell mitotic activity is an important aspect of muscle development. An understanding of the interplay between nutrition and satellite cell mitotic activity will lead to more efficient meat production. The objective of this study was to test the influence of the leucine metabolite, beta-hydroxy beta-methylbutyrate (HMB), and feed deprivation on muscle development in the early posthatch poult. Male Nicholas poults were placed on 1 of 4 treatments: immediately fed a starter diet with 0.1% HMB (IF-HMB), immediately fed a starter diet containing 0.1% Solka-Floc for a control (IF-No HMB), feed and water withheld for 48 h immediately posthatch and then fed the HMB diet (WF-HMB), and feed and water withheld for 48 h immediately posthatch and then fed the control starter diet (WF-No HMB). 5-bromo-2'-deoxyuridine (BrdU) was injected intra-abdominally into all poults to label mitotically active satellite cells. The pectoralis thoracicus was harvested 2 h after the BrdU injection. Immunohistochemistry for BrdU, Pax7, and laminin along with computer-based image analysis was used to study muscle development. IF-HMB poults had higher body weights (P < 0.01) at 48 h and 1 wk of age and had higher satellite cell mitotic activity at 48 h of age (P < 0.01) compared with the IF-No HMB and WF poults. Therefore, dietary supplementation of HMB may have an anabolic effect on early posthatch muscle.

Maintenance of glucose homeostasis during late-term embryonic development is dependent upon the amount of glucose held in reserve primarily in the form of glycogen in the liver and upon the degree of glucose generated by gluconeogenesis from protein first mobilized from amnion albumen and then from muscle. Insufficient glycogen and albumen will force the embryo to mobilize more muscle protein toward gluconeogenesis, thus restricting growth of the late-term embryo and hatchling. We hypothesize that administration of available carbohydrates to the amnion will improve glycogen reserves and spare muscle protein mobilization for gluconeogenesis during late-term embryonic and posthatch neonatal development. Our hypothesis was tested by comparing BW gain, liver glycogen reserves, and muscle weight of in ovo fed and control embryos during last days of embryonic incubation until 25 d after hatching. We examined, using 600 birds from 2 different strains of commercial boilers, body and muscle weights and glycogen reserves following feeding embryos at d 17.5 of incubation with a solution containing maltose, sucrose, dextrin, and beta-hydroxy-beta-methylbutyrate (HMB). Providing carbohydrates and HMB to late-term embryos increased hatching weights by 5 to 6% over controls, improved liver glycogen by 2- to 5-fold, and elevated relative breast muscle size by 6 to 8%. These weight advantages were sustained through the end of the experiments at 25 d of age. It is reasonable to assume that the elevated glycogen levels in the in ovo treatment reduce the need to produce glucose via gluconeogenesis and, therefore, contribute to less use of muscle protein and hence a greater percentage of pectoral muscle weight in the in ovo birds.

Early development of the digestive tract is crucial for achieving maximal growth and development of chickens. Because the late-term embryo naturally consumes the amniotic fluids, insertion of a nutrient solution into the embryonic amniotic fluid [in ovo (IO) feeding] may enhance development. This study examined the effect of IO feeding on d 17.5 of incubation of carbohydrates (CHO) and beta-hydroxy-beta-methylbutyrate (HMB) on small intestinal development of chickens during the pre and posthatch periods. Results shows that 48 h post-IO feeding procedure all IO feeding treatments exhibited increased villus width and surface area compared with the control group. At d 3 posthatch the surface area of an average villi was increased by 45% for the HMB IO group and by 33% for the CHO and CHO+HMB IO groups compared with controls (noninjected fertile eggs). The activity of jejunal sucrase-isomaltase (SI) was higher (P < 0.05) 48 h after IO feeding in all the IO fed embryos, whereas at day of hatch and at d 3 the CHO+HMB IO group had the highest maltase activity (P < 0.05), which was approximately 50% greater than control embryos. These observations indicated that small intestines of IO fed hatchlings were functionally at a similar stage of development as a conventionally fed 2-d-old chick. Body weight of all IO fed hatchlings was greater than controls, and these differences (P < 0.05) were sustained until the end of the experiment (10 d). At d 10 chicks that were IO fed with CHO had BW that were 2.2% higher, whereas HMB and CHO+HMB IO fed chicks showed 5 to 6.2% BW increase, respectively, compared with controls. The current study shows that the administration of exogenous nutrients into the amnion enhanced intestinal development by increasing the size of the villi and by increasing the intestinal capacity to digest disaccharides. This advantage probably leads to higher BW in IO fed chicks.

As a dietary supplement, beta-Hydroxy-beta-Methylbutyrate (HMB), a catabolite of leucine, has been shown to reduce broiler mortality. In a series of experiments, male broilers (Experiments 1 and 2, n = 576) were grown for 21 days on diets that contained HMB at 0, 0.01. 0.05, and 0.10% of diet. In Experiment 3 (n = 240), chicks were fed diets containing 0, 0.05, 0.075, and 0.10% HMB. HMB dietary supplementation did not significantly affect broiler weight gain in any experiment. However, a trend toward increased mean broiler weight gain per bird was observed in Experiments 1 and 3 when HMB was consumed at 0.10% of the diet. Mean feed to gain ratio was not affected by the inclusion of HMB in broiler diets. In Experiment 3, HMB supplemented diets did not affect bursa of Fabricius, thymus, and spleen weights at 21 days of age. Cutaneous basophilic hypersensitivity response against pokeweek mitogen was higher (P < or = 0.05) at 48 and 72 hours post-injection in chicks on 0.05% dietary HMB (Experiment 1). In Experiment 2, this increase occurred 24 hours post-injection in chicks fed HMB at 0.01% of the diet. On the contrary, the T-cell mediated response against PHA-P mitogen was comparable between all dietary treatments in multiple experiments. Macrophage function profiles were determined at 21 days of age. All chicks in experiments 1 and 2 on HMB supplemented diets showed an increase in the recruitment of Sephadex-G50-elicited abdominal exudate cells (AEC). A 2-fold increase in AEC numbers occurred at the 0.10% HMB level (Experiment 1, P < or = 0.05). Although HMB supplementation did not significantly affect the phagocytic potential of the abdominal macrophages, nitrite levels in the macrophage culture supernatants were higher in 0.01% and 0.05% treatment groups as compared to the controls (Experiment 2, P < or = 0.04; Experiment 3, P < or = 0.05). HMB supplementation did not alter the bird's ability to clear Escherichiacoli or Salmonella arizona from the bloodstream. Beginning 7 days post-hatch, chicks were injected i.v. with a 7% sheep red blood cells suspension. Serum samples were collected to determine the primary and secondary antibody response. Chicks receiving the 0.1% HMB diet in Experiments 1 and 2 exhibited increased IgG and total anti-sheep red blood cell (SRBC) antibody levels during the primary response. During the secondary response, birds consuming the 0.10% HMB diet had elevated IgM levels as well as increased total anti-SRBC levels over the controls in Experiments 1 and 3. These studies show that HMB supplementation improves several immunological functions in young broilers, and such improvement may result in decreased mortality.

Beta-hydroxy-beta-methylbutyrate (HMB), a leucine catabolite, has been shown to decrease broiler mortality. One possible target of HMB action may be the cells of the immune system. Macrophages from a chicken macrophage cell line, MQ-NCSU, were exposed to 0, 10, 20, 40, 80, and 100 microg of HMB per 5 x l0(4) cells in a 96-well culture plate. After 24 h of exposure, macrophage proliferation was quantitated by an MTT bioassay. In duplicate experiments, HMB stimulated growth over control (p < or = 0.05) at a wide range of doses. Macrophages were exposed to 20 and 80 microg of HMB and the culture supernatant fractions tested for the presence of nitrite. HMB exposure (20 microg) increased nitrite production by 44.1% over the controls (Experiment 1, p< or =0.035). To determine the phagocytic potential of macrophages after HMB exposure, MQ-NCSU cell line and Sephadex-G50-elicited abdominal macrophages were incubated with fluorescent latex beads (1:40, macrophage to beads ratio) for I h and then analyzed by flow cytometry. When exposed to 40 microg HMB, the phagocytic potential of MQ-NCSU macrophages was significantly higher (31.7%) than that of the controls (p < or = 0.0006). Sephadex-elicited macrophages exhibited 14.4% increased phagocytosis over controls when treated with 80 microg HMB (p < or = 0.0016). When MQ-NCSU macrophages were exposed to HMB, Fc-receptor expression was significantly elevated over the controls (p < or = 0.0001). These data demonstrate that HMB exposure induces proliferation of macrophages in culture as well as enhances macrophage effector functions, such as nitrite production and phagocytosis. The findings of these studies imply that HMB can be used as a possible dietary immunomodulator.

Experiments were conducted to determine whether the leucine catabolite beta-hydroxy-beta-methylbutyrate (HMB) could influence protein metabolism in broilers. In Experiment 1, HMB was fed at either .01 or .05% of the diet. beta-Hydroxy-beta-methylbutyrate did not improve feed conversion or BW gain; however, broilers fed HMB at .01% in a diet formulated to meet NRC (1984) recommendations had no mortality (P < .03) from 0 to 21 d of age. In Experiment 2, HMB fed at .003, .01, .03, and .09% of the diet had no significant affect on growth or carcass yield of the broilers when compared with control-fed broilers. In Experiment 3, HMB fed at .001, .003, and .01% of the diet had no effect on broiler growth. In Experiment 4, HMB was fed at .01% of the diet in combination with an antibiotic and coccidiostat (GP). Use of GP increased (P < .01) BW, feed conversion, and carcass yield when compared with the control broilers. In Experiment 5, HMB was fed at .1% of the diet, and effects of sex and GP were examined. beta-Hydroxy-beta-methylbutyrate decreased (P < .01) mortality by 72% in the male broilers. In a combined analysis, HMB fed at .01% of the diet (Experiments 1, 2, and 3) increased breast yield (P < .05) and reduced mortality by 56% (P < .04) from 0 to 21 d of age. In a combined analysis, HMB fed at .003% of the diet (Experiments 2 and 3) increased 42-d BW (P < .02) and hot (P < .04) and chilled (P < .05) carcass yields. In conclusion, across all HMB dosages mortality of male broilers was decreased from 6.37 to 4.39% (-31%, P < .04) by feeding HMB, with the pattern of death suggesting that HMB decreased the incidence of sudden death syndrome in these broilers.