Impact of dichromatic lighted incubation on hatching result and post-hatch performance of broiler chickens.

This study was planned to evaluate the impact of dichromatic lights during incubation on the hatching and post-hatch performance of broiler chickens. A total of 500 eggs of broiler breeder (Ross 308; Age 44 weeks) were evenly divided according to a completely randomized design into 4 treatments having 5 replicates and 25 eggs each. Treatments consisted of dichromatic lights Blue + Red (BR), Green + Red (GR) and Green + Blue (GB) provided at an intensity of 250 lx for 12 h a day along with a Dark (D) environment. After hatching 200 chicks (50 from each respective light group) were divided into 4 treatments with 5 replicates each having 10 chicks. Results indicated a higher embryo index (13.12%) in the GR group on the 12th day of incubation; while an ideal hatch window was observed in GR and GB (98.18% and 96.00% hatched chicks) lighting groups. In hatching traits, higher hatchability (86.15) and hatch of fertile (93.85) percentages were observed in GR lighting followed by GB, BR and Dark treatment groups; while dead-in shell embryos were lowest in the GR group. In growth performance, higher feed intake (513.20 g) and body weight (479.20 g) were observed in the GB group followed by GR, BR and dark group; and feed conversion ratio (FCR) was better in the GR group (1.06). In welfare parameters, improved physical asymmetry (0.90 mm) and tonic immobility (54.40 s) were measured in the GR group followed by GB, BR and the dark group. It was concluded that under experimental conditions when broiler breeder eggs are provided with GR lighting during incubation, it can help to improve hatchability, growth performance and welfare traits in chicks.

Eggshell translucency: its relationship with specific gravity and eggshell color and its influence on broiler egg weight loss, hatchability, and embryonic mortalities.

Eggshell quality is among the most important factors affecting hatchability in broiler breeders, and therefore several methods for its assessment are available in the poultry industry. Among them, eggshell translucency has received special attention in recent years due to its connection with ultrastructural disorganization of the shell layers. However, there is very limited data on the impact of translucency on hatching eggs and on the possible links between this trait and specific gravity (SG) or shell color. Thus, our study investigated associations and interactions between eggshell translucency, SG, and color on incubation parameters of eggs from the same breeding flock (Ross 308AP, 51 wk of age). To this end, light and dark eggs within 5 different SG categories (≥1.065, 1.070, 1.075, 1.080, and ≤1.085) were selected from 15,976 eggs, graded into 3 translucency scores, and later incubated to evaluate egg weight loss, hatchability and embryonic mortalities. In general, translucency scores were evenly distributed within SG categories (χ2 [8, N = 1,138] = 13.67, P = 0.090) and color (χ2 [2, N = 1,138] = 4.93, P = 0.084). No interactions between eggshell translucency and SG or between translucency and color were found for the analyzed variables. An interaction was observed between SG and eggshell color for the variable egg weight loss, where the light-shelled eggs, in most SG categories lost more weight throughout incubation than dark eggs. Eggshell translucency affected egg weight loss, hatchability, and embryonic mortality on 11 to 18 d of incubation, with highly translucent eggs showing the worst results. At the same time, eggs with SG lower than 1.070 displayed the greatest weight loss, lowest hatchability, and highest contamination. We found no influence of eggshell color on weight loss or hatchability, but light-shelled eggs exhibited higher late embryonic mortality. Together, these data suggest that despite its effects on certain hatching parameters, shell translucency bears no relationship to SG or color.

Differential physiological response of slow- and fast-growing broiler lines to hypoxic conditions during chorioallantoic membrane development.

Ambient conditions during chicken embryogenesis, such as insufficient oxygen or changes in temperature, are expected to cause permanent phenotypic changes and affect their posthatch performance. Decades of genetic selection for high growth rate resulted with various physiological and morphological changes that can affect the broiler fitness under environmental stress. To evaluate the selection effect on responses to environmental challenge during embryonic development, and the long-term implications, we have used a unique genetic line, that was not selected for over 30 yr (since 1986), as control for the modern commercial genetic line. At embryonic day 5 (E5), broiler embryos from these 2 genetic lines were divided into 2 treatments: 1) control; 2) 15% O2 concentration for 12 h/day from E5 through E12 the embryonic period of chorioallantoic membrane formation. Embryos and hatched chicks were characterized for physiological and morphological parameters. Significant differences in relative embryo weight and yolk consumption were found between the 2 lines. The modern line was characterized by a higher metabolic rate and rapid growth, supported by higher hemoglobin levels and hematocrit concentrations, whereas the 1986 line had slower metabolism, lower levels of hematocrit and hemoglobin, higher oxygen volume per 1 g of embryonic tissue indicating higher oxygen availability. Both lines exhibited changes in heart rate, and blood parameters corresponding to cardiovascular system adaptation after hypoxic exposure, seemingly implemented to increase oxygen-carrying capacity to the embryo tissues. Our finding stand in agreement that the genetic selection for high growth rate that led to higher metabolism without a fit of the cardiovascular system, increased the imbalance between oxygen supply and demand.

Results of hatching and rearing broiler chickens in different incubation systems.

Hatchery efficiency is based on hatchability and the number of salable chicks. The hatchery sector has been seeking new alternatives to optimize production rates, including the use of different systems (multistage [MS] or single-stage [SS] machines) to improve incubation conditions. The present study aimed to compare results for hatchability, chick quality, and broiler performance of chicks from 2 incubator systems-MS and SS. The experimental design for hatchability, hatch window, egg weight loss, and chick performance variables was completely randomized with 2 treatments (MS and SS). Performance variables were analyzed as a 2 x 2 factorial arrangement (incubator type x chick sex). Egg weight loss between incubation and transfer was higher for eggs incubated in MS (P < 0.05). Hatchability was higher for eggs incubated in SS (P < 0.05), and chicks in SS had a longer hatch window (P < 0.05). Embryo diagnosis revealed higher final mortality for embryos incubated in MS (P < 0.05), as well as higher percentages of alive and dead pipped and cracked eggs (P < 0.05). Physical quality was better for chicks from SS (P < 0.05). There was no interaction between the studied factors for performance results (P > 0.05). Incubator type did not affect broiler performance for any of the studied ages (P > 0.05), whereas male broilers had better performance than females (P < 0.05). The SS incubation system proved better than the MS system at meeting embryo requirements during embryo development, with better hatching rates and chick quality, although performance variables were not influenced by incubation type.

Transcriptomic and Epigenetic Preservation of Genetic Sex Identity in Estrogen-feminized Male Chicken Embryonic Gonads.

Whereas in ovo exposure of genetically male (ZZ) chicken embryos to exogenous estrogens temporarily feminizes gonads at the time of hatching, the morphologically ovarian ZZ-gonads (FemZZs for feminized ZZ gonads) are masculinized back to testes within 1 year. To identify the feminization-resistant "memory" of genetic male sex, FemZZs showing varying degrees of feminization were subjected to transcriptomic, DNA methylome, and immunofluorescence analyses. Protein-coding genes were classified based on their relative mRNA expression across normal ZZ-testes, genetically female (ZW) ovaries, and FemZZs. We identified a group of 25 genes that were strongly expressed in both ZZ-testes and FemZZs but dramatically suppressed in ZW-ovaries. Interestingly, 84% (21/25) of these feminization-resistant testicular marker genes, including the DMRT1 master masculinizing gene, were located in chromosome Z. Expression of representative marker genes of germline cells (eg, DAZL or DDX4/VASA) was stronger in FemZZs than normal ZZ-testes or ZW-ovaries. We also identified 231 repetitive sequences (RSs) that were strongly expressed in both ZZ-testes and FemZZs, but these RSs were not enriched in chromosome Z. Although 94% (165/176) of RSs exclusively expressed in ZW-ovaries were located in chromosome W, no feminization-inducible RS was detected in FemZZs. DNA methylome analysis distinguished FemZZs from normal ZZ- and ZW-gonads. Immunofluorescence analysis of FemZZ gonads revealed expression of DMRT1 protein in medullary SOX9+ somatic cells and apparent germline cell populations in both medulla and cortex. Taken together, our study provides evidence that both somatic and germline cell populations in morphologically feminized FemZZs maintain significant transcriptomic and epigenetic memories of genetic sex.

Chick Embryo Growth Modeling Using Near-Infrared Sensor and Non-Linear Least Square Fitting of Egg Opacity Values.

Non-destructive monitoring of chick embryonic growth can provide vital management insights for poultry farmers and other stakeholders. Although non-destructive studies on fertility, hatching time and gender have been conducted recently, there has been no available method for embryonic growth observation, especially during the second half of incubation. Therefore, this work investigated the feasibility of using near-infrared (NIR) sensor-based egg opacity values-the amount of light lost when passing through the egg-for indirectly observing embryo growth during incubation. ROSS 308 eggs were selected based on size, mass and shell color for this experiment. To estimate the embryo size precisely, we fit various mathematical growth functions during incubation, based on the opacity value of incubated eggs. Although all the growth models tested performed similarly in fitting the data, the exponential and power functions had better performances in terms of co-efficient of determination (0.991 and 0.994 respectively) and RMSE to explain embryo growth during incubation. From these results, we conclude that the modeling paradigm adopted provides a simple tool to non-invasively investigate embryo growth. These models could be applied to resolving developmental biology, embryonic pathology, industrial and animal welfare issues in the near future.

Effects of gonadotropins on testis cell subpopulations of newly hatched chicks treated during embryonic development.

The aim of the present study was to evaluate histological and stereological changes, as well as the variations in the number and size of cells from diverse cell subpopulations in testes of newly hatched chicks treated with follicle stimulating hormone (FSH) and luteinizing hormone (LH) during embryonic development. Stereological results indicated that in FSH-treated chicks total volume of the tubular compartment constitutes most of the testis. In contrast, the total volume of interstitial tissue constitutes most of the testis of LH-treated chicks. Results indicate that the number of germ and Sertoli cells increases as a result of FSH and LH treatment, but in FSH-treated testis, Sertoli cells were the most numerous cell type in seminiferous tubules; whereas germ cells were the most numerous cell type in testis of LH-treated chicks. Results also indicate there was a larger total volume of Leydig cells in the testes of FSH- and LH-treated chicks. The larger volume of Leydig cells in FSH-treated chicks is due to a larger cellular volume of these cells, and not due to the number, which remains constant. In contrast, in testes of LH-treated chicks, there is a larger number and volume of Leydig cells. These results indicate the testes of chick embryos respond to FSH and LH treatment, with there being modifications in the seminiferous tubules and interstitial tissue, but these changes differ markedly, indicating that FSH and LH have differential effects on chick testes.

Effect of in ovo application of hydroxyapatite nanoparticles on chicken embryo development, oxidative status and bone characteristics.

Intensive selection in modern lines of fast-growing chickens has caused several skeletal disorders. Therefore, current research is focused on methods to improve the bones of birds. A new potential solution is in ovo technology using nanoparticles with a high specificity for the bone tissue. Thus, the objective of the present study was to evaluate the effect of in ovo application of hydroxyapatite nanoparticles (HA-NP) in different concentrations (50, 100 and 500 µg/ml colloids) on chicken embryo development, with a particular focus on the oxidative status and bone characteristics of the embryo. The results showed that in ovo treatment with HA-NP did not negatively affect hatchability and body weight. However, bone weight was reduced in 500 µg/ml group. The concentrations of calcium, phosphorus and crude ash were not affected. The modulatory effect of HA-NP was observed on the basis of antioxidative markers - superoxide dismutase, total antioxidant status, malondialdehyde in serum and selected tissues. Glutathione concentration in serum suggested higher metabolic stress. Among bone turnover markers, the concentration of osteocalcin was found to be significantly affected by HA-NP injection. Thus, the in ovo application of HA-NP could modify the molecular responses at the stage of embryogenesis but these changes were not reflected in embryo growth and even slowed down bone development. Nevertheless, the question for the follow-up research is whether in ovo administration of HA-NP would affect the antioxidative status and bone turnover resulting in improved bone conditions and body gain in post hatch chickens.

Effects of stage of broiler embryo development on coccidiosis vaccine injection accuracy, and subsequent oocyst localization and hatchling quality.

Control of coccidiosis in broiler chickens continues to pose challenges to commercial poultry producers, especially in an era of increased consumer demand for antibiotic-free broiler production. As a result, coccidiosis vaccines are now commonly used in rotation programs to achieve effective coccidiosis control. Inovocox EM1 vaccine (EM1) is a coccidiosis vaccine that allows for earlier immune acquisition through oocyst cycling, which reduces the effects of wild-type coccidia. The EM1 vaccine is administered to embryonated broiler hatching eggs between 18 and 19 D of incubation (doi). In the U.S., commercial broiler hatcheries vaccinate embryonated eggs at either 18.5 or 19 doi. However, it is unclear whether a difference in embryo age at the time of in ovo injection can impact the actual site of vaccine delivery. In addition, it is unclear where oocysts eventually become localized within the embryo following the in ovo injection of EM1. Therefore, the objective of this study was to determine the effects of stage of embryonic development on the actual deposition site of the EM1 vaccine oocysts when they are in ovo injected and to subsequently investigate the movement and eventual location of EM1 oocysts after in ovo injection. Because all eggs were injected at the same time, a 12-h difference in set time was a means to derive 18.5 and 19.0 incubation age of injection (IAN) treatments. The experimental design was a 3 injection treatment (noninjected, diluent-injected, and vaccine-injected) × 2 IAN factorial. There was a significant main effect of IAN on site of vaccine oocysts delivery, and subsequent hatching chick quality. Qualitative histological evaluation revealed the oral uptake of vaccine oocysts through the amnion, with their subsequent presence in the gizzard and intestinal lumen by 24 to 36 h postinjection. In conclusion, physiological development influenced the site of injection, and oocysts imbibed along with the amniotic fluid in late stage broiler embryos are subsequently transported to the gastrointestinal tract.

Maternal conjugated linoleic acid alters hepatic lipid metabolism via the AMPK signaling pathway in chick embryos.

The effects of maternal conjugated linoleic acid (CLA) on embryonic development and hepatic lipid metabolism were investigated in chick embryos. A total of 180 Arbor Acres female broiler breeders (36 wk old) were randomly divided into the following 3 dietary treatment groups: a basic diet (control), a basic diet containing 0.5% CLA (CLA1), and a basic diet containing 1.0% CLA (CLA2). The females were fed for 8 wk, and the eggs from each group were collected and hatched during the last 2 wk. The results showed that the addition of dietary CLA increased the broken egg rate and reduced the fertilization rate and the egg hatchability (P < 0.05). CLA enrichment decreased the polyunsaturated and monounsaturated fatty acids and increased the saturated fatty acids in the yolk sac (P < 0.05). The yolk sac weight, body weight, and body length had a linear decrease with CLA supplementation (P < 0.05). In the developing chick embryo (at E14) and newly hatched chick (D0), the serum triglyceride concentration decreased with maternal CLA supplementation and was accompanied by a reduction in subcutaneous adipose tissue deposition. In addition, maternal CLA supplementation mediated the hepatic lipid metabolism by decreasing the mRNA expression of sterol regulatory element-binding proteins-1c (SREBP-1c), fatty acid synthase and acetyl-CoA carboxylase, and increasing the mRNA expression of adenosine 5'-monophosphate-activated protein kinase α (AMPKα), peroxisome proliferator-activated receptors α (PPARα), liver fatty acid-binding protein, adipose triglyceride lipase and carnitine palmitoyltransferase in embryonic chick livers (P < 0.05). A drop in SREBP-1c protein expression and an increase in the protein expression of p-AMPKα and PPARα were also observed in the liver of chick embryo (P < 0.05). In conclusion, maternal CLA supplementation regulated the fatty acid composition in the yolk sac, and mediated embryonic chick development and hepatic lipometabolism, and these effects may be related to the AMPK pathway. These findings suggest the potential ability of maternal CLA supplementation to reduce fat deposition in chick embryos.

Dynamic morphoskeletons in development.

Morphogenetic flows in developmental biology are characterized by the coordinated motion of thousands of cells that organize into tissues, naturally raising the question of how this collective organization arises. Using only the kinematics of tissue deformation, which naturally integrates local and global mechanisms along cell paths, we identify the dynamic morphoskeletons behind morphogenesis, i.e., the evolving centerpieces of multicellular trajectory patterns. These features are model- and parameter-free, frame-invariant, and robust to measurement errors and can be computed from unfiltered cell-velocity data. We reveal the spatial attractors and repellers of the embryo by quantifying its Lagrangian deformation, information that is inaccessible to simple trajectory inspection or Eulerian methods that are local and typically frame-dependent. Computing these dynamic morphoskeletons in wild-type and mutant chick and fly embryos, we find that they capture the early footprint of known morphogenetic features, reveal new ones, and quantitatively distinguish between different phenotypes.

Proteomic analysis of fertilized egg yolk proteins during embryonic development.

Egg yolk is an important source of nutrients for embryo development. In this study, the egg yolk protein composition at 0, 10, and 18 D of incubation was analyzed by 2-dimensional gel electrophoresis (2-DE) combined with matrix-assisted laser desorption/ionization time-of-flight tandem mass spectrometry. A significant difference in the abundance of 42 protein spots representing 12 proteins were identified (P < 0.05). The 2-DE gel image analysis exhibited that the molecular weight (MW) of 29 protein spots was lower than their theoretical value, in which 14 vitellogenin (VTG) fragments were lower than the theoretical value. There were 13 protein spots showed a higher MW including 5 ovotransferrins with MW of 87.2 kDa. The gene ontology enrichment analysis suggested that biological process of the differentially expressed proteins were mainly involved in lipid transport and lipid localization at 10 and 18 D of incubation. The molecular function of the differentially expressed proteins was involved in nutrient reservoir activity, lipid transporter activity, and antigen binding at 10 D of incubation. At 18 D of incubation, the differentially expressed proteins mainly participated in nutrient reservoir activity and substrate-specific transporter activity. The high abundance of VTGs at 10 D of incubation might participate in lipid localization and lipid transportation to facilitate the yolk nutrient transport to embryo. The low expression of ovotransferrins at 10 D of incubation indicated the chondrogenesis of embryo.

Preliminary evaluation of application of a 3-dimensional network structure of siloxanes Dergall preparation on chick embryo development and microbiological status of eggshells.

The spatial network structure of Dergall is based on substances nontoxic to humans and the environment which, when applied on solid surfaces, creates a coating that reduces bacterial cell adhesion. The bacteriostatic properties of siloxanes are based on a purely physical action mechanism which excludes development of drug-resistant microorganisms. The aims of the present study were to 1) evaluate a Dergall layer formed on the eggshell surface regarding the potential harmful effects on the chick embryo; 2) evaluate antimicrobial activity and estimate the prolongation time of Dergall's potential antimicrobial activity. Dergall at a concentration of 0.6% formed a layer on the eggshell surface. In vitro testing of the potential harmful effects of Dergall by means of a hen embryo test of the chorioallantoic membrane showed no irritation reaction at a concentration of 3% and lower. The hatchability of the groups sprayed with a Dergall water solution with a concentration of 0 to 5% was 89.1 to 93.8% for fertilized eggs (P > 0.05) but decreased to 63.7% (P < 0.05) in the group sprayed with a 6% concentration of the solution. This phenomenon was caused by embryo mortality in the first week of incubation. At the concentration of 0.6%, Dergall exhibited strong antibacterial properties against bacteria such as Staphylococcus aureus, Escherichia coli, Shigella dysenteriae, Shigella flexneri, and Salmonella typhimurium. For Streptococcus pyogenes, the highest antibacterial activity of Dergall was reported in the concentrations of 100 and 50%. For Pseudomonas aeruginosa, no antibacterial activity of Dergall was generally observed, but in vivo testing showed a strong decrease of all gram-negative bacteria growth. Moreover, a prolonged antimicrobial effect lasting until 3 D after disinfection was observed, which makes Dergall a safe and efficient disinfectant.

Structure, function, and evolution of Gga-AvBD11, the archetype of the structural avian-double-ß-defensin family.

Out of the 14 avian ß-defensins identified in the Gallus gallus genome, only 3 are present in the chicken egg, including the egg-specific avian ß-defensin 11 (Gga-AvBD11). Given its specific localization and its established antibacterial activity, Gga-AvBD11 appears to play a protective role in embryonic development. Gga-AvBD11 is an atypical double-sized defensin, predicted to possess 2 motifs related to ß-defensins and 6 disulfide bridges. The 3-dimensional NMR structure of the purified Gga-AvBD11 is a compact fold composed of 2 packed ß-defensin domains. This fold is the archetype of a structural family, dubbed herein as avian-double-ß-defensins (Av-DBD). We speculate that AvBD11 emanated from a monodomain gene ancestor and that similar events might have occurred in arthropods, leading to another structural family of less compact DBDs. We show that Gga-AvBD11 displays antimicrobial activities against gram-positive and gram-negative bacterial pathogens, the avian protozoan Eimeria tenella, and avian influenza virus. Gga-AvBD11 also shows cytotoxic and antiinvasive activities, suggesting that it may not only be involved in innate protection of the chicken embryo, but also in the (re)modeling of embryonic tissues. Finally, the contribution of either of the 2 Gga-AvBD11 domains to these biological activities was assessed, using chemically synthesized peptides. Our results point to a critical importance of the cationic N-terminal domain in mediating antibacterial, antiparasitic, and antiinvasive activities, with the C-terminal domain potentiating the 2 latter activities. Strikingly, antiviral activity in infected chicken cells, accompanied by marked cytotoxicity, requires the full-length protein.

Integrated Proteomic and N-Glycoproteomic Analyses of Chicken Egg during Embryonic Development.

To better appreciate the alterations of egg proteins and their modifications during embryonic development, a comparative and quantitative study was performed aimed at chicken egg white and yolk proteome and N-glycoproteome after 12 days of incubation using tandem mass tag (TMT)-labeling technology in conjunction with reversed-phase high-performance liquid chromatography (RP-HPLC). A total of 334 unique N-glycosite-containing peptides from 153 N-glycoproteins were identified, of which 82 N-glycosite-containing peptides showed significant changes after 12 days of incubation. The varied proteome was mainly involved with antibacterial, ionic binding, cell proliferation, and embryonic development, while the different degrading and/or absorbing priorities of egg proteins were proposed. This study provides substantial insight into the effects of N-glycoprotein variations on the utilization of egg proteins by chicken embryo during incubation.

Immune responses upon in ovo HVT-IBD vaccination vary between different chicken lines.

The genotype of chickens is assumed to be associated with variable immune responses. In this study a modern, moderate performing dual-purpose chicken line (DT) was compared with a high-performing layer-type (LT) as well as a broiler-type (BT) chicken line. One group of each genotype was vaccinated in ovo with a recombinant herpesvirus of turkeys expressing the virus protein VP2 of the infectious bursal disease virus (HVT-IBD) while one group of each genotype was left HVT-IBD unvaccinated (control group). Genotype associated differences in innate and adapted immune responses between the groups were determined over five weeks post hatch. HVT-IBD vaccination significantly enhanced humoral immune responses against subsequently applied live vaccines compared to non-HVT-IBD vaccinated groups at some of the investigated time points (P < 0.05). In addition HVT-IBD vaccination had depending on the genotype a significant impact on splenic macrophage as well as bursal CD4+ T-cell numbers (P < 0.05). On the other hand, the detectable genotype influence on Interferon (IFN) γ and nitric oxide (NO) release of ex vivo stimulated spleen cells was independent of HVT-IBD vaccination. The results of our study suggest considering a genotype specific vaccination regime in the field.

Light-dark rhythms during incubation of broiler chicken embryos and their effects on embryonic and post hatch leg bone development.

There are indications that lighting schedules applied during incubation can affect leg health at hatching and during rearing. The current experiment studied effects of lighting schedule: continuous light (24L), 12 hours of light, followed by 12 hours of darkness (12L:12D), or continuous darkness (24D) throughout incubation of broiler chicken eggs on the development and strength of leg bones, and the role of selected hormones in bone development. In the tibiatarsus and femur, growth and ossification during incubation and size and microstructure at day (D)0, D21, and D35 post hatching were measured. Plasma melatonin, growth hormone, and IGF-I were determined perinatally. Incidence of tibial dyschondroplasia, a leg pathology resulting from poor ossification at the bone's epiphyseal plates, was determined at slaughter on D35. 24L resulted in lower embryonic ossification at embryonic day (E)13 and E14, and lower femur length, and lower tibiatarsus weight, length, cortical area, second moment of area around the minor axis, and mean cortical thickness at hatching on D0 compared to 12L:12D especially. Results were long term, with lower femur weight and tibiatarsus length, cortical and medullary area of the tibiatarsus, and second moment of area around the minor axis, and a higher incidence of tibial dyschondroplasia for 24L. Growth hormone at D0 was higher for 24D than for 12L:12D, with 24L intermediate, but plasma melatonin and IGF-I did not differ between treatments, and the role of plasma melatonin, IGF-I, and growth hormone in this process was therefore not clear. To conclude, in the current experiment, 24L during incubation of chicken eggs had a detrimental effect on embryonic leg bone development and later life leg bone strength compared to 24D and 12L:12D, while the light-dark rhythm of 12L:12D may have a stimulating effect on leg health.

The effect of increased concentration of carbon dioxide during the first 3 days of incubation on albumen characteristics, embryonic mortality and hatchability of broiler hatching eggs1.

Three experiments were conducted to determine the effects of increased CO2 concentration during the first 3 d of incubation on albumen height and pH, embryonic mortality, and hatchability of broiler hatching eggs. Hatching eggs were obtained from commercial broiler breeder flocks of Ross 308 at 39 and 37 wk of age in Experiments 1 and 2, respectively. In Experiment 3, eggs were collected at 28 and 35 wk of age. Eggs were incubated under either standard conditions (Control-CO2) for the entire incubation or increased CO2 concentrations during the first 3 d of incubation (High-CO2) in 3 experiments. In Experiments 1 and 2, the CO2 concentration was gradually increased from the beginning of incubation onwards to reach 0.80% at 72 h by manual injection of CO2 into airtight laboratory incubators. In the control incubators, the CO2 concentration remained below 0.10% during the same period. Prior to setting, and at 3 d of incubation, the eggs were opened for albumen height and pH measurements in Experiments 1 and 2. In Experiment 3, the eggs were set in commercial incubators. During the first 3 d of incubation, the CO2 concentration was gradually increased to reach 0.70% at 72 h naturally (High-CO2). In the Control-CO2 incubator, the CO2 concentration remained below 0.10%. After 3 d, incubation was continued with the control incubator conditions for all eggs from both groups in the 3 experiments. The albumen height was not affected by CO2 treatment, but the treatment significantly decreased albumen pH at 3 d in Experiments 1 and 2 (P < 0.05). A greater CO2 concentration during early incubation reduced fertile hatchability due to increased early embryonic mortality by 2% in the 3 experiments (P ≤ 0.05). The differences in pH might provide one explanation why increased CO2 concentration during early incubation resulted in increased early embryonic mortality. These data indicated that at the beginning of the incubation, ventilation was necessary to prevent increases in CO2 concentration for optimum hatchability results.

Organotypic Culture Method to Study the Development Of Embryonic Chicken Tissues.

The embryonic chicken is commonly used as a reliable model organism for vertebrate development. Its accessibility and short incubation period makes it ideal for experimentation. Currently, the study of these developmental pathways in the chicken embryo is conducted by applying inhibitors and drugs at localized sites and at low concentrations using a variety of methods. In vitro tissue culturing is a technique that enables the study of tissues separated from the host organism, while simultaneously bypassing many of the physical limitations present when working with whole embryos, such as the susceptibility of embryos to high doses of potentially lethal chemicals. Here, we present an organotypic culturing protocol for culturing the embryonic chicken half head in vitro, which presents new opportunities for the examination of developmental processes beyond the currently established methods.

Transcriptional profiling of liver during the critical embryo-to-hatchling transition period in the chicken (Gallus gallus).

BACKGROUND: Although hatching is perhaps the most abrupt and profound metabolic challenge that a chicken must undergo; there have been no attempts to functionally map the metabolic pathways induced in liver during the embryo-to-hatchling transition. Furthermore, we know very little about the metabolic and regulatory factors that regulate lipid metabolism in late embryos or newly-hatched chicks. In the present study, we examined hepatic transcriptomes of 12 embryos and 12 hatchling chicks during the peri-hatch period-or the metabolic switch from chorioallantoic to pulmonary respiration. RESULTS: Initial hierarchical clustering revealed two distinct, albeit opposing, patterns of hepatic gene expression. Cluster A genes are largely lipolytic and highly expressed in embryos. While, Cluster B genes are lipogenic/thermogenic and mainly controlled by the lipogenic transcription factor THRSPA. Using pairwise comparisons of embryo and hatchling ages, we found 1272 genes that were differentially expressed between embryos and hatchling chicks, including 24 transcription factors and 284 genes that regulate lipid metabolism. The three most differentially-expressed transcripts found in liver of embryos were MOGAT1, DIO3 and PDK4, whereas THRSPA, FASN and DIO2 were highest in hatchlings. An unusual finding was the "ectopic" and extremely high differentially expression of seven feather keratin transcripts in liver of 16 day embryos, which coincides with engorgement of liver with yolk lipids. Gene interaction networks show several transcription factors, transcriptional co-activators/co-inhibitors and their downstream genes that exert a 'ying-yang' action on lipid metabolism during the embryo-to-hatching transition. These upstream regulators include ligand-activated transcription factors, sirtuins and Kruppel-like factors. CONCLUSIONS: Our genome-wide transcriptional analysis has greatly expanded the hepatic repertoire of regulatory and metabolic genes involved in the embryo-to-hatchling transition. New knowledge was gained on interactive transcriptional networks and metabolic pathways that enable the abrupt switch from ectothermy (embryo) to endothermy (hatchling) in the chicken. Several transcription factors and their coactivators/co-inhibitors appear to exert opposing actions on lipid metabolism, leading to the predominance of lipolysis in embryos and lipogenesis in hatchlings. Our analysis of hepatic transcriptomes has enabled discovery of opposing, interconnected and interdependent transcriptional regulators that provide precise ying-yang or homeorhetic regulation of lipid metabolism during the critical embryo-to-hatchling transition.