In ovo feeding with minerals and vitamin D3 improves bone properties in hatchlings and mature broilers.

The objective of this study was to examine the effect of in ovo feeding (IOF) with inorganic minerals or organic minerals and vitamin D3 on bone properties and mineral consumption. Eggs were incubated and divided into 4 groups: IOF with organic minerals, phosphate, and vitamin D3 (IOF-OMD); IOF with inorganic minerals and phosphate (IOF-IM); sham; and non-treated controls (NTC). IOF was performed on embryonic day (E) 17; tibiae and yolk samples were taken on E19 and E21. Post-hatch, only chicks from the IOF-OMD, sham, and NTC were raised, and tibiae were taken on d 10 and 38. Yolk mineral content was examined by inductively coupled plasma spectroscopy. Tibiae were tested for their whole-bone mechanical properties, and mid-diaphysis bone sections were indented in a micro-indenter to determine bone material stiffness (Young's modulus). Micro-computed tomography (µCT) was used to examine cortical and trabecular bone structure. Ash content analysis was used to examine bone mineralization. A latency-to-lie (LTL) test was used to measure standing ability of the d 38 broilers. The results showed that embryos from both IOF-OMD and IOF-IM treatments had elevated Cu, Mn, and Zn amounts in the yolk on E19 and E21 and consumed more of these minerals (between E19 and E21) in comparison to the sham and NTC. On E21, these hatchlings had higher whole-bone stiffness in comparison to the NTC. On d 38, the IOF-OMD had higher ash content, elevated whole-bone stiffness, and elevated Young's modulus (in males) in comparison to the sham and NTC; however, no differences in standing ability were found. Very few structural differences were seen during the whole experiment. This study demonstrates that mineral supplementation by in ovo feeding is sufficient to induce higher mineral consumption from the yolk, regardless of its chemical form or the presence of vitamin D3. Additionally, IOF with organic minerals and vitamin D3 can increase bone ash content, as well as stiffness of the whole bone and bone material in the mature broiler, but does not lead to longer LTL.

Differences in intestinal mucin dynamics between germ-free and conventionally reared chickens after mannan-oligosaccharide supplementation.

A germ-free (GF) chicken model was used to test 2 hypotheses: 1. microbial colonization of the gastrointestinal tract (GIT) influences mucin gene expression and mucin types; and 2. mannan oligosaccharide (MOS) supplementation affects GIT cells directly, without bacteria mediation, compared with bacterial-mediated effect (i.e., indirectly). Gnotobiotic isolators were used: 1) GF, 2) with a single bacteria population, and 3) conventionalized by exposure to cecal bacterial contents. Each was divided to 2 diet groups: with or without MOS (2 kg/t) for 1 wk. Results show that the absence of bacteria in the GIT caused a reduction in neutral and acidic goblet cell (GC) number and density, an increase in sulfated mucin, absence of sialylated GC, and reduced mucin 2 mRNA expression in the small intestine of GF compared with conventional birds. These results indicate a reduced development of mucin production and secretion in the absence of GIT bacteria implying a less mature small intestine mucosa, supporting our first hypothesis. Results from the single bacteria population group were not conclusive and did not support any of the hypotheses. Supplementation of MOS, regardless of microbial presence, caused a reduction in neutral GC number and density but increased neutral GC area. The MOS caused different effects on acidic mucins in conventional and GF birds, causing a reduction in sialylated GC number (conventional) and a reduction in sulfated GC density (GF), all supporting a direct effect of MOS in GF animals, in addition to an indirect effect via gut microflora.

Dietary guanidinoacetate reduces spaghetti meat myopathy risk in the breast muscle of broiler chickens.

The global demand for white chicken meat along with the increase in the occurrence of growth-related breast muscle myopathies (BMMs) [namely white striping (WS), wooden breast (WB), and spaghetti meat (SM)] highlights the need for solutions that will improve meat quality while maintaining the high productivity of modern broilers. Guanidinoacetate (GAA), a precursor of creatine, is used as a feed additive and has previously shown the potential to affect the quality of breast meat. This study investigated growth performance, meat quality and the risk ratio for the development of BMMs in broilers assigned to two dietary treatments: control (CON) group, fed a commercial basal diet, and supplemented GAA (sGAA) group, receiving the control diet supplemented on top with 0.06% GAA. Growth performance indicators such as BW, daily weight gain, daily feed intake, feed conversion ratio and cumulative feed conversion ratio were recorded on a pen basis. As a trait affecting animal welfare, the occurrence of foot pad dermatitis was also evaluated. At day 43, birds were processed, and breasts were scored for the incidence and severity of BMMs (n = 166 and 165 in CON and sGAA groups, respectively). Quality traits (ultimate pH, colour) and technological properties (i.e., drip and cooking losses, marinade uptake, shear force, and oxidation levels of the lipid and the protein fractions) of breast meat were assessed in both treatments on samples not showing any macroscopic sign of BMMs (n = 20 breast fillets per group). Data of myopathy risk ratio were analysed as the risk for each group to develop WS, WB, and SM myopathies. Our results show that while sGAA and control groups did not differ significantly in growth performance, a remarkably beneficial effect of GAA was observed on the incidence of BMMs with significantly reduced risk of sGAA group to develop SM myopathy. The risk of sGAA group to develop SM was 30% lower compared to CON (P = 0.028). Finally, a significantly lower drip loss was observed in sGAA in comparison with CON (1.78 vs 2.48%, P = 0.020). Together, our results show that the inclusion of 0.06% GAA in feed can improve the water-holding capacity of meat and reduce the risk to develop SM myopathy without compromising the performance of broilers.

Changes in yolk sac membrane absorptive area and fat digestion during chick embryonic development.

The capacity of yolk sac (YS) utilization by the chick embryo may be affected by structural changes in the YS membrane (YSM) and by the mechanisms within its cells for digestion, absorption, and transfer of nutrients. Two experiments were conducted to examine structural and digestive changes in the YS of the broiler chick embryo; weights of embryo, YS, and YSM, as well as the total area of the YSM and the absorptive area of the YSM, were measured between embryonic day (E) 5 and E21. In addition, fat content, lipase activity, and bile acid concentration in the YSM and YS contents (YSC) were measured between E11 and E21. Results showed that YSM weight increased from 0.19 g on E5 to 6.46 g on E15, and decreased by 3.74 g between E17 and E21. The absorptive YSM area increased from 536 mm² on E5 (51% of total YSM area) to 6,370 mm² (86% of total area) on E17, and decreased to 4,439 mm(2) on E21 (85% of total area). The smaller YSM area between E17 and E21 did not decrease the rate of YS fat utilization, which could suggest that YSM mechanisms for fat absorption, digestion, and secretion increased during that period. Total YSM lipase activity relative to fat content (units per g of YSM fat) increased from approximately 1,000 units on E15 to 1,500 units on E21. The detection of lipase in the YSM lends support to the hypothesis that YS lipids are hydrolyzed in the lipolysosomes of the YSM. The current study also confirmed for the first time that bile acids are present in the YS, with levels that ranged from 0.61 to 1.06 µmol/g in the YSM, and may suggest that bile is synthesized in the YSM of the chick embryo. Results of the current study contribute to our understanding of the developmental changes that affect YS functionality and could give insight into the coordination between the embryo's demands and YSM morphological, absorptive, digestive, and secretive changes.

Metabolic profiling of late-term turkey embryos by microarrays.

The last stages of embryonic development are crucial for turkeys as their metabolism shifts to accommodate posthatch survival and growth. To better understand the metabolic change that occurs during the perinatal period, focused microarray methodology was used to identify changes in the expression of key genes that control metabolism of turkey embryos from 20 d of incubation (E) until hatch (E28). Gene expression patterns were evaluated in liver, pectoral muscle, and hatching muscle and were associated with measured embryonic growth and tissue glycogen concentration. Within the studied period, the expression of 60 genes significantly changed in liver, 53 in pectoral muscle, and 51 in hatching muscle. Genes related to lipid metabolism (enoyl-CoA hydratase, 3-hydroxyacyl-CoA dehydrogenase, 3-hydroxymethylglutaryl-CoA reductase, acetyl-CoA carboxylase, lipoprotein lipase, and thyroxine deiodinase) had reduced expression between E22 and E26, corresponding to the period of expected limited oxygen supply. In contrast, genes related to opposing pathways in carbohydrate metabolism, such as glycolysis and gluconeogenesis (hexokinases, glucose-6 phosphatase, phosphofructokinases, glucose 1-6 phosphatase, pyruvate kinase, and phosphoenolpyruvate carboxykinase), or glycogenesis and glycogenolysis (glycogen synthase and glycogen phosphorylase) had rather static expression patterns between E22 and E26, indicating their enzymatic activity must be under posttranscriptional control. Metabolic survey by microarray methodology brings new insights into avian embryonic development and physiology.

Effects of dietary carbohydrates on rumen epithelial metabolism of nonlactating heifers.

Ruminal wall metabolism was studied in nonlactating heifers by altering the carbohydrate (CHO) digestion site between rumen and intestine. The CHO digestion site was estimated from in situ and total-tract digestibility of control (CONT) diets and diets supplemented with corn (CRN), barley (BARL), or soy hulls (SOYH). Ruminal epithelial metabolism regulating gene expression, morphology, and nutrient delivery was assessed from a combination of rumen volatile fatty acid (VFA) concentration, biopsies for papilla morphology, and expression of putative metabolic regulatory genes encoding enzymes that facilitate VFA utilization. Digestible dry matter and CHO intake were 25 and 45% higher, respectively, in the supplemented diets than in CONT diets. Fiber supplementation increased the intestinal and decreased ruminal CHO digestion. Ruminal nonfiber CHO digestibility was 10% lower in CRN than with the high rumen-degradable supplement. The CONT heifers had lowest total ruminal VFA and highest acetate concentration relative to the other treatments. Total VFA concentration in BARL and CRN diets tended to be higher than in SOYH. The SOYH diet tended to reduce papilla dimension relative to CRN and BARL. The CRN diet tended to increase papilla surface area relative to BARL and SOYH. Gene expression of propionyl-coenzyme A carboxylase was higher in CRN and BARL than in SOYH diets, and tended to be higher in CRN than in BARL and SOYH diets. Lactate dehydrogenase and butyryl coenzyme A synthase gene transcripts tended to be higher in CONT than in the supplemented treatments. Thus, rumen epithelial expression of genes involved in VFA metabolism and ruminal wall-structure development are influenced by other regulatory mechanism that is not directly affected by local signals. The in situ methods used are a useful tool for differentiating ruminal from extraruminal nutrient supply.

Yolk sac carbohydrate levels and gene expression of key gluconeogenic and glycogenic enzymes during chick embryonic development.

Glycogen and glucose concentrations (mg/g of tissue) and amounts (mg) were determined in the yolks of fertile eggs on the day of set and in the yolk sac (YS) and liver of broiler chick embryos between 11 and 21 embryonic days of age (E). On the day of set, the yolk contained 50 mg of glucose (0.31% of yolk) but did not contain glycogen. During incubation, the amount of glucose in the YS increased from 20 mg on E11 to 60 mg on E19. A parallel increase in YS and liver glycogen concentrations (mg/g) during the last week of incubation implied a similar capacity for glycogen synthesis per gram of tissue. However, due to its larger size, the YS capacity for glycogen storage far exceeded that of the liver, which stored less than 12 mg of glycogen up to E19, as compared with more than 200 mg in the YS. Between E19 and 21, liver and YS glycogen amounts decreased by 10 mg and 100 mg, respectively. These results indicated that the YS is a glycogenic and perhaps gluconeogenic organ. We therefore evaluated the gene expression of glycogen synthase and glycogen phosphorylase as well as gluconeogenic enzymes (fructose 1,6-bisphosphatase, phosphoenolpyruvate carboxykinase, and glucose 6-phosphatase) in the YS membrane and liver by real-time reverse-transcription PCR. Although the YS membrane and liver displayed different patterns of mRNA abundance, the high abundance of fructose 1,6-bisphosphatase mRNA in the YS membrane between E11 and 15, and the expression of phosphoenolpyruvate carboxykinase and glucose 6-phosphatase, supported the postulated gluconeogenic abilities of the YS membrane and indicated its role in providing glucose to the embryo. Thus, glucose is probably synthesized in the YS, stored in the form of glycogen, and toward hatch, the YS may have the potential to transfer 10 times more glycogen-derived glucose to the embryo as compared with the liver. As such, the YS may play a major role in the synthesis and storage of glucose and its supply to the chick embryo toward hatch.

Bone characteristics of late-term embryonic and hatchling broilers: bone development under extreme growth rate.

The development of broilers is an extreme example of rapid growth, increasing in weight from 40 g at hatch to 2,000 g 5 to 6 wk later. Such rapid growth requires a correspondingly fast development of the skeleton. Bone development is a genetically programmed process that is modified by epigenetic factors, mainly muscle-induced stresses and strains. In this study, we describe the temporal changes in bone morphology and material properties during the prehatch period [embryonic day (E) 14, E17, E19, E21] and posthatch d 3 and 7. The bones were examined for their weight, length, ash content, mechanical properties, and cortical structure. We show that the cross-sectional shape of the tibia and femur changes during the examination period from circular to elliptical. Additionally, the changes in bone properties are time-dependent and nonuniform: from E14 to E17 and from d 3 to 7, fast bone growth was noted, with major increases in both mechanical properties (stiffness, ultimate load, and energy to fracture) and geometric properties (cross-sectional area and thickness, medullary area, and moment of inertia). On the other hand, during the last days of incubation, most mechanical and geometric properties remain unchanged or even decrease. The reasons for this finding may relate to the hatching process but also to mineral shortage during the last days of incubation. This study leads to better understanding of bone development in ovo and posthatch in fast-growing broilers.

Gene expression of nutrient transporters and digestive enzymes in the yolk sac membrane and small intestine of the developing embryonic chick.

Chick embryos derive nutrients from the yolk during incubation and transition to intestinal absorption of nutrients posthatch. The uptake of nutrients is mediated by a variety of membrane-bound transporter proteins. The objective of this study was to determine the expression profiles of nutrient transporters and digestive enzymes during incubation in the yolk sac membrane (YSM) and embryonic intestine of egg-laying (Leghorn) and meat-producing (Cobb) chickens derived from 22 to 30 wk (young) and 45 to 50 wk (old) breeder flocks. Transporters examined included the peptide transporter PepT1, the glutamate/aspartate (EAAT3), cationic (CAT-1) and neutral (B0AT) amino acid transporters, and the fructose (GLUT5) and glucose (SGLT1) transporters. Digestive enzymes included aminopeptidase N (APN) and sucrase-isomaltase (SI). Expression of these genes was assessed by real-time PCR using the absolute quantification method in YSM at embryonic day (E) 11, 13, 15, 17, 19, 20, and 21 and intestine at E15, 17, 19, 20, and 21. The PepT1 and APN gene expression in the YSM increased until E15 and then decreased until E21, whereas expression in the intestine increased from E15 to E21. The B0AT showed a similar pattern, with greatest expression in the YSM occurring at E17/E19. The CAT1 and GLUT5 genes showed decreased expression in the YSM and increased expression in the intestine until E17/E19 and then a decrease until E21. Expression of SGLT1 and EAAT3 showed increased gene expression over time in both the intestine and YSM. Expression of SI showed little to no gene expression in the YSM, whereas the intestine exhibited consistently high levels of gene expression. In YSM and intestine, SI expression was greater in Leghorn than Cobb, whereas CAT1 and GLUT5 expression was greater in Cobb than Leghorn. Expression of the APN, CAT1, and SI genes was greater in embryos from young flocks than old flocks in YSM and intestine. These results demonstrate that the YSM expresses many of the digestive enzymes and nutrient transporters typically associated with the intestine and that these genes show tissue- and development-specific patterns of expression.

Content and uptake of minerals in the yolk of broiler embryos during incubation and effect of nutrient enrichment.

Although embryo and chicken growth and development rely on mineral nutrition, information on mineral levels in the egg compartments during incubation is limited. Accordingly, we examined P, Ca, Fe, Zn, Cu, and Mn levels in the yolk of breeder eggs during incubation and the effect of embryonic mineral (with specific nutrients) enrichment on yolk mineral levels and consumption. First, fertile eggs were examined on day of setting (DOS), embryonic day (E) 11, E13, E15, E17, E19, E20, and day of hatch (DOH) for the mineral content in the yolk (and albumen on DOS) by inductively coupled plasma atomic emission spectroscopy. Results showed that on DOS, the yolk is the major origin for Mn, P, Fe, Ca, Cu, and Zn. Interestingly, P, Fe, Zn, Cu, and Mn were mostly consumed from the yolk until E17, after which their consumption was very low. Consumption of P was constant until E17 and then decreased until E20. Consumption of Fe, Zn, Cu, and Mn was medium to mild until E11, increased between E11 and E17, and minimal between E17 and DOH. Enrichment treatment, where fertile eggs were divided into 2 groups [nonenriched (control) and enriched (with minerals, vitamins, and carbohydrates on E17 using the in ovo feeding method)] showed that the enriched group had higher Fe, Zn, Cu, and Mn levels than the nonenriched group and exhibited higher consumption of Fe, Zn, and Mn between E20 and DOH. Analysis of the shell mineral composition along incubation showed that the shell released low amounts of P, Fe, and Mn in comparison with the yolk mineral content. Therefore, we concluded that the shell is a minor source of these minerals. Studying the mineral resources and consumption of embryos can lead to a better understanding of the mineral limitations of embryos during incubation. Additionally, because minerals are important for the development of the embryo, the higher mineral levels and consumption observed in the enriched group may affect the development of critical organs, such as the skeletal system.

The chick embryo yolk sac membrane expresses nutrient transporter and digestive enzyme genes.

The yolk sac membrane plays a major role in the transport of nutrients from the yolk contents to the chick embryo. We examined whether the yolk sac membrane expresses genes for nutrient digestion, enzymes, and nutrient transporters. We evaluated relative mRNA abundance of the digestive enzymes aminopeptidase N (APN) and sucrase-isomaltase (SI); the nutrient transporters oligopeptide transporter Pept1, cationic amino acid transporter CAT1, and sodium glucose transporter SGLT1; and the micronutrient transporters type IIb sodium phosphate cotransporter NPT2b, calcium transporter TRPV6, and zinc transporter ZnT-1 from embryonic d 11 (11E) to 21E (day of hatch) by real-time reverse-transcription PCR. The yolk sac membrane expressed all the examined genes, which exhibited several patterns of expression. Relative abundance of APN mRNA increased in the yolk sac membrane from 11E to 17E and decreased from 17E to 20E. Expression of PepT1 increased from 11E to 15E and decreased from 15E to 20E. In contrast, CAT1 expression decreased from 11E to 13E and increased from 15E to 17E. Expression of SGLT1 increased between 15E and 20E and decreased substantially between 20E and 21E. Expression of NPT2b increased during incubation and exhibited the highest relative expression of all the examined genes, particularly on 20E to 21E. Expression of TRPV6 decreased from 11E to 13E and increased substantially from 15E to 19E. No significant difference was found between the sampled days for ZnT-1 or SI expression, with the latter exhibiting the lowest relative expression of all the genes studied. These results present the first documentation of nutrient transporter and digestive enzyme gene-expression patterns in the yolk sac membrane, and provide a basis for future research on the capacity of the yolk sac membrane for nutrient digestion and transport.

Effect of in ovo feeding and its interaction with timing of first feed on glycogen reserves, muscle growth, and body weight.

Chicks are commonly fasted for the first 36 to 72 h posthatch because of the logistics of commercial production. Fasting for 48 to 72 h posthatch results in retarded BW, delayed intestinal development, and lower pectoral muscle weight. This study is focused on the first 36 h of fasting and its interaction with feeding before hatch. Four treatment groups, differing in time of first feed, 6 h [early feeding (EF)] or 36 h [standard feeding procedure (SP)] posthatch, with or without in ovo feeding (IOF) with dextrin and ß-hydroxy-ß-methylbutyrate-calcium salt in a saline solution, were examined for glycogen status in the liver and pectoral muscle, myogenic cell proliferation, and myofiber diameter in embryos and chickens on various days posthatch. In addition, chicken BW, ADG, pectoral muscle weight, and pectoral muscle percentage of BW until 35 d of age were recorded. Results showed that delaying the first feed for 36 h posthatch (SP group) led to an irreversibly reduced growth rate compared with the EF group. However, IOF affected the growth of chickens in the SP group, whereas the control embryos had depleted glycogen reserves in the liver; IOF-treated embryos had elevated hepatic glycogen contents on embryonic day (E) 19, E20, and the day of hatch. In addition, on d 2 posthatch, although hatchlings in the SP group showed the predicted low levels of glycogen in their livers, birds in the EF group exhibited more than 30-fold and 3-fold increases in liver and muscle glycogen, respectively. In ovo-fed birds in the SP group also exhibited higher glycogen reserves, BW, pectoral muscle weight, and BW gain than control birds in the SP group. In ovo feeding had an immediate effect on promoting myoblast proliferation on E19, whereas on d 3 posthatch, the effect was pronounced only in the EF groups. On d 5, although myoblast proliferation in all groups declined, it remained higher in both IOF groups. These effects were expressed on d 3 and 35 by myofiber diameter. Together, IOF had a long-term supportive effect on BW and posthatch muscle growth when first feed was delayed by 36 h.

The effect of in ovo administration of mannan oligosaccharide on small intestine development during the pre- and posthatch periods in chickens.

Early intestinal development is essential for chicken embryos to fulfill their maximal growth potential. Mannan oligosaccharide (MOS) is known to improve gut morphology, function, and innate immunity; therefore, we hypothesized that its administration in the prehatch period to the sterile intestine of embryos would affect intestinal development and functionality without mediation of gut microflora. The MOS was administered by in ovo feeding procedure to embryos 3 d before hatch. the effects of MOS administration on intestinal morphology, activity of the brush-border enzymes amino peptidase (AP) and sucrase isomaltase (SI) and mRNA abundance of AP, SI, sodium-dependent glucose cotransporter 1 (SGLT1), peptide transporter 1 (PepT1), secreted mucin (MUC2), and toll-like receptors (TLR2 and TLR4) were examined and compared with saline-injected and noninjected controls. Results show that on embryonic d 20 the only parameter affected was MUC2 mRNA abundance, which exhibited a 3-fold increase in the MOS group versus controls. On day of hatch more parameters were affected: a 20 to 32% increase in villus area was found in the MOS group compared with controls; crypt depth and number of goblet cells per villus were higher by 20 and 50%, respectively, compared with the saline group; and AP and SI activities were higher by 44 and 36%, respectively, compared with the noninjected control. In addition, an increase in fold change mRNA abundance of AP, SI, and TLR4 was observed in the MOS group compared with controls. However, on d 3 posthatch, a decrease in MOS effects was noted, indicating a temporally limited effect after administration of 1 dose. In ovo administration of MOS prehatch resulted in a hatching chick with more mature enterocytes and enhanced epithelial barrier and digestive and absorptive capacity at day of hatch. Results imply that the mechanism underlying the observed changes is not mediated through gut microflora but rather involves a direct effect of MOS on intestinal cells.

Centennial Review: The chicken yolk sac is a multifunctional organ.

The yolk sac (YS) consists of the yolk, which supplies nutrients, and the YS tissue, which surrounds the yolk and provides essential metabolic functions for the developing embryo. The YS tissue is derived from the midgut of the embryo and consists of a layer of endodermal epithelial cells (EEC) in contact with the yolk contents, a mesodermal layer that contains the vascular system and an outer ectodermal layer. The YS tissue is a multifunctional organ that provides essential functions such as host immunity, nutrient uptake, carbohydrate and lipid metabolism, and erythropoiesis. The YS tissue plays a role in immunity by the transport of maternal antibodies in the yolk to the embryonic circulation that feeds the developing embryo. In addition, the YS tissue expresses high mRNA levels of the host defense peptide, avian ß-defensin 10 during mid embryogenesis. Owing to its origin, the YS EEC share some functional properties with intestinal epithelial cells such as expression of transporters for amino acids, peptides, monosaccharides, fatty acids, and minerals. The YS tissue stores glycogen and expresses enzymes for glycogen synthesis and breakdown and glucogenesis. This carbohydrate metabolism may play an important role in the hatching process. The mesodermal layer of the YS tissue is the site for erythropoiesis and provides erythrocytes before the maturation of the bone marrow. Other functions of the YS tissue involve synthesis of plasma proteins, lipid transport and cholesterol metabolism, and synthesis of thyroxine. Thus, the YS is an essential organ for the growth, development, and health of the developing embryo. This review will provide an overview of the studies that have investigated the functionalities of the YS tissue at the cellular and molecular levels with a focus on chickens.

Yolk sac nutrient composition and fat uptake in late-term embryos in eggs from young and old broiler breeder hens.

In the present study, we examined the composition, amount, and uptake of yolk nutrients [fat, protein, water, and carbohydrates (COH)] during incubation of eggs from 30- and 50-wk-old broiler breeder hens. Eggs were sampled at embryonic d 0 (fresh eggs), 13, 15, 17, 19, and 21 (hatch). Egg, embryo, yolk content, and yolk sac membrane were weighed, and the yolk sac (YS; i.e., yolk content + yolk sac membrane) composition was analyzed. From 30 to 50 wk of age, the albumen weight increased by 13.3%, whereas the yolk increased by more than 40%. The proportion of fat in the fresh yolk of the 30-wk-old group was 23.8% compared with 27.4% in the 50-wk-old group, whereas the proportion of protein was 17.9% compared with 15.6%, respectively. During incubation, results indicated that water and protein infiltrated from other egg compartments to the YS. Accordingly, the calculated change in the content of water and protein between fresh yolk and sampled YS does not represent the true uptake of these components from the YS to the embryo, and only fat uptake from the YS can be accurately estimated. By embryonic d 15, fat uptake relative to embryo weight was lower in the 30-wk-old group than in the 50-wk-old group. However, by embryonic d 21, embryos of both groups reached similar relative fat uptake, suggesting that to hatch, embryos must attain a certain amount of fat as a source of energy for the hatching process. The amount of COH in the YS increased similarly during incubation in eggs from hens of both ages, reaching a peak at embryonic d 19, suggesting COH synthesis in the YS. At hatch, the amount of protein, water, and COH in the residual YS, relative to the weight of the yolk-free chick, was similar in eggs from young and old hens. However, chicks from the younger hens had less fat in the YS for their immediate posthatch nutrition compared with those from the older hens.

Prehatch intestinal maturation of turkey embryos demonstrated through gene expression patterns.

Some of the challenges faced by neonatal turkeys include weakness, reduced feed intake, impaired growth, susceptibility to disease, and mortality. These symptoms may be due to depleted energy reserves after hatch and an immature digestive system unable to replenish energy reserves from consumed feed. To better understand enteric development in turkeys just before hatch, a new method was used to identify the patterns of intestinal gene expression by utilizing a focused microarray. The duodenums of 24 turkey embryos were sampled on embryonic day (E)20, E24, E26, and hatch (E28). The RNA populations of 96 chosen genes were measured at each time point, from which 81 significantly changed (P < 0.01). These genes were clustered by gene expression pattern similarity into 4 groups. The expression pattern of hormone receptors revealed that intestinal tissues may be less responsive to growth hormone, insulin, glucagon, and triiodothyronine during the last 48 h before hatch, when developmental emphasis switches from cell proliferation to functional maturation. Based on gene expression patterns, we concluded that at hatch, poults should have the capacity to 1) digest disaccharides but not oligopeptides, due to increased expression of sucrase-isomaltase but decreased expression of aminopeptidases and 2) absorb monosaccharides and small peptides due to high expression of sodium-glucose cotransporter-4 and peptide transporter-1.

The effects of in ovo feeding arginine, beta-hydroxy-beta-methyl-butyrate, and protein on jejunal digestive and absorptive activity in embryonic and neonatal turkey poults.

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.

The effect of heat stress on ovarian function of laying hens.

Reproductive failure associated with heat stress is a well-known phenomenon. The mechanism involved in this failure is not clearly understood. In order to test a possible direct effect of heat stress on ovarian function, 36 White Leghorn laying hens were housed in individual cages in 2 temperature- and light-controlled rooms (n = 18). At 31 wk of age, one group was exposed daily for 12 h to high temperature (42 +/- 3 degrees C), and the second group was maintained under thermoneutral conditions (24 to 26 degrees C) and served as control. Body temperature, feed intake, egg production, and egg weight were recorded daily; heparinized blood samples were drawn every 3 d for plasma hormonal level of luteinizing hormone, follicular stimulating hormone, progesterone, 17beta-estradiol, and testosterone. Six days after exposure half of the birds in each group were killed, and the ovary and oviduct were weighed and preovulatory follicles removed and extracted for mRNA of Cytochrome P 450 aromatase, 17-alpha hydroxylase. The same procedure was repeated 9 d later with the rest of the birds. Short and long heat exposure caused significant hyperthermia and reduction of egg production, egg weight, ovarian weight, and the number of large follicles. In addition, a significant reduction in plasma progesterone and testosterone was detected 2 d after exposing the birds to heat stress, and plasma 17beta-estradiol was significantly reduced 14 d after initiation of heat stress. Short exposure to heat stress caused significant reduction in mRNA expression of cytochrome P450 17-alpha hydroxylase, exposing the birds to long-term heat stress caused significant reduction in expression of mRNA of both steroidogenic enzymes. No significant change was found in plasma luteinizing hormone and follicular stimulating hormone levels during the entire experimental period. We suggest a possible direct effect of heat stress on ovarian function.

The role of the bursa of Fabricius in the immune response to vaccinal antigens and the development of immune tolerance in chicks (Gallus domesticus) vaccinated at a very young age.

Vaccination is recognized to be the most cost-effective means of preventing, controlling, and even eradicating infectious diseases. Conventional poultry are vaccinated through various routes including eye/nose drops, drinking water, vent brush, or injection. Efficient vaccination is an essential part of any good poultry management.The bursa of Fabricius is intimately connected to the cloaca and the intestinal system. It is well-known as a primary lymphoid organ in the chicken and a major channel through which environmental antigens stimulate the immune system. In this study we tested whether direct instillation of various viral vaccines and antigens into the cloaca (per bursam), could stimulate higher antibody titers and generate improved protection. Despite the very rapid absorption of the vaccines or antigens from the cloaca to the lumen of the Bursa of Fabricius, per bursam inoculation failed to generate a satisfactory immune response. In contrast conventional administration of live or inactivated commercial vaccines led to an acceptable level of seroconversion and protection against challenge.An interesting finding in this study was the fact that administration of a single priming dose of antigenic material at age 1 or 5 days, did not improve the response to a second administration at 14 days of age as expected. Instead, in most cases there was a reduced serum antibody response suggesting the induction of tolerance. This was true for all routes of administration (intramuscular, per ocular and per bursam) and for all formulations of vaccine.The current study reveals: 1) no advantage for direct application of live or inactivated vaccines or antigens into the bursa of Fabricius compared to common routes of vaccination, 2) that apparent desensitization or tolerance effects have important implications for poultry management, since in many countries, vaccination of day old chicks is compulsory or a well-accepted part of flock vaccination.According to our results, early vaccination can in fact reduce or inhibit a secondary immune response to subsequent vaccination and increase susceptibility to disease agents.

Maternal and genetic effects on broiler bone properties during incubation period.

In order to examine the differences in bone properties between fast-growing and slow-growing broiler embryos and to understand the effects of genotype and egg size on these differences, fast- and slow-growing hens and males were reciprocally crossed to create 4 egg groups: FST (laid by fast-growing hens, inseminated by fast-growing males), H-FST (fast-growing hens and slow-growing males), H-SLW (slow-growing hens and fast-growing males), and SLW (slow-growing hens and slow-growing males). Embryos (n = 8) from these 4 groups were sacrificed and weighed, and both tibiae were harvested on embryonic d (E) 17, 19, and 21. Left tibiae were tested for their whole-bone mechanical properties using a micromechanical device. Cortical bone structure and bone mineral density (BMD) were examined by micro-computed tomography of the left tibiae. Bone mineralization was evaluated by measuring BMD and ash content, while the rate and location of mineralization were evaluated by fluorochrome labeling. Osteoclastic activity and osteocyte density were evaluated by histological stains [TRAP (Tartrate resistant acid phosphatase) and H&E (Hematoxylin and Eosin), respectively]. Groups with larger eggs (FST and H-FST) had higher BW and tibia weight than groups with smaller eggs (SLW and H-SLW); however, they had a lower ratio of tibia weight to BW. Between groups with similar egg weight, stiffness, maximal load, and yield load of the bones were higher in the SLW than the H-SLW, while no differences were found between the FST and H-FST. Additionally, the tibiae of the SLW were stiffer and their osteocyte density higher than in the FST on E21 and their periosteal mineralization rate was higher between E19 and E21. No differences were found between the groups in cortical bone structure. This study demonstrates that faster growing hatchlings, especially those that hatch from relatively small eggs, have inferior bone mechanical properties in comparison to slower growing hatchlings, and suggests that fast-growing chicks hatching from small eggs are at a higher risk for developing bone pathologies. Accordingly, selection for increased egg size may lead to improved mechanical performance of the skeleton of fast-growing broilers.