In ovo feeding of vitamins in broilers: A comprehensive meta-analysis of hatchability and growth performance.

In ovo feeding has been introduced as a cost-effective method to improve hatchability and broiler performance. Specifically, several studies have focused on the impact of vitamins. However, due to variations in experimental conditions across all trials, drawing general conclusions appears challenging. Therefore, we conducted a meta-analysis of 17 published papers, including a maximum of 134 sample size to evaluate the potential effects of in ovo feeding of vitamins in broilers. Studies were retrieved by consulting scientific repositories such as Pubmed, Scopus, Scielo, Web of Science, and Google Scholar. A binary logistic model was used to determine the parameters influencing hatchability. To assess variations in hatchling weight and growth parameters based on the vitamin category, a mixed model analysis of variance was performed, considering the study as a random effect and the vitamin category as a fixed effect. Finally, a linear mixed model was used to develop equations that explain the evolution of growth parameters based on vitamin concentration, volume, and day of injection. The results revealed that for better hatchability, it is preferable to consider heavier eggs (p = 0.007), lower volumes (p = 0.039), and late injection (p = 0.022). Vitamin E was associated with higher hatchling weight (p = 0.037), while vitamin C exhibited the lowest overall feed conversion ratio (p = 0.042). Interactions were observed between the day of injection and vitamin concentration or volume of injection for all studied growth parameters. In summary, the findings of this study suggest that hatchability during in ovo feeding is influenced by technique-related parameters, whereas growth parameters can be modulated by the category of vitamin injected. Consequently, this study lays the groundwork for future investigations assessing the effects of in ovo feeding in broilers, as it highlights the relationship between the methodology and potential outcomes.

Expression of nutrient transporter genes in response to dietary rice gluten meal and protease enzyme supplementation and the consequent effects on growth, nutrient digestibility, immunity and jejunum histomorphometry in chicken.

The objective of this study was to investigate the effects of feeding rice gluten meal (RGM) as an alternative protein source along with protease enzyme supplementation on growth performance, expression of nutrient transporter genes, nutrient digestibility, immune response and gut histomorphometry of broiler chicken. Proximate analysis of RGM revealed 923 g dry matter (DM), 500 g crude protein (CP), 69.2 g ether extract, 94.7 g crude fiber, 215.4 g nitrogen-free extract, 43.7 g ash, 6.20 g calcium, 7.80 g total phosphorus, 18.99 MJ gross energy and 12.68 MJ metabolizable energy per kg diet. Significant upregulation of nutrient transporter genes (PepT1, EAAT3 and mucin) and better growth performance was observed in the birds fed control diet which was statistically similar to the birds fed 150 g RGM compared to birds fed higher RGM levels. Histomorphometry of jejunum, nutrient digestibility, and immune response of birds did not reveal any significant effect of RGM or protease enzyme supplementation. However, the inclusion of RGM up to 150 g/kg diet resulted in significant decline of feed cost/kg live weight gain, dressed meat yield and eviscerated meat yield by 13.13%, 12.99% and 13.36%, respectively compared to control. Thus, it was concluded that the inclusion of 150 g RGM/kg diet in broiler chicken ration has no adverse effects on the growth pattern of birds and can be used for least-cost feed formulation for chicken.

Effect of thermal conditioning on growth performance and thermotolerance in broilers: A systematic review and meta-analysis.

Thermal conditioning has been introduced as a cost-effective way to improve performance and thermotolerance in broilers. However, since all the trials were performed under various experimental conditions, it appears difficult to draw general conclusions. Therefore, the objective of this study was to quantify the response of broilers to thermal conditioning through a meta-analysis approach. A literature search was conducted on Scopus, PubMed, Scielo, Web of Science, and Google scholar in December 2020. A restricted maximum likelihood random effect model was used to pool the effect sizes from the body weight gain (BWG), feed intake (FI), feed conversion ratio (FCR), and body temperature (Tb). BWG, FI, and Tb were computed using the standardized mean difference (SMD) while FCR was computed using mean differences (MD) with a 95% confidence interval (IC). Growth performances were evaluated during the thermoneutral conditions while Tb was evaluated after either acute or chronic heat stress after early age thermal conditioning. A total of 17 studies were included in the dataset. Thermal conditioning significantly increased BWG (SMD = 0.139, IC = 0.0372-0.2407, P = 0.0074) and FI (SMD = 0.292, IC = 0.108-0.476, P = 0.0019) compared with the control. Additionally, subgroup analysis revealed that overall Tb was significantly reduced under acute heat stress (SMD = -0.455, IC = -0.718 to -0.192, P < 0.001) but not affected during chronic heat stress (SMD = -0.115, IC = -0.651 to -0.420, P = 0.6729). In conclusion, thermal conditioning significantly increased the broiler's BWG and FI under thermoneutral conditions and can help in reducing Tb under acute heat stress.

Heat stress management in poultry.

High ambient temperature is one of the major causes of economic losses in the livestock industry. The poultry industry is an integral part of the livestock industry. It faces severe losses due to heat stress (HS). The adverse effects of HS can be seen on production performance, body temperature, intestinal health, appetite hormone regulation, immune responses and oxidative characteristics. It is important to monitor these parameters to identify the HS possessions during rearing so that timely action can be taken to minimize the adverse effects of high ambient temperature. Furthermore, the application of productive methods on farms is equally important. Several strategies have been suggested by researchers. Providing a suitable environment with selective rearing systems along with proper ventilation and hygiene is the basic requirement for all types of livestock reared for animal protein. Supplementation of appropriate feed additive could be useful for improving intestinal absorption and minimizing adverse effects of HS. Selection for breeding heat resistant birds also provide merits for improving the germplasm of the strains. Early age thermal conditioning also helps in developing resistance for HS. The most recent advancement is the supplementation of active substances during incubation. It is expected that these methods may have a potential impact on the poultry industry for creating thermotolerance in the newly hatched chicks. This review highlights the major issues concerning chicken health and suggests the measures to be adopted following the increase in environmental temperature.

Dynamics of gene expression of hormones involved in the growth of broiler chickens in response to the dietary protein and energy changes.

The synergy between the genetic potential and the nutrient intake determines the growth performance of meat-type chicken and nutrigenomics approach helps us understand the response of candidate genes of growth in chicken to dietary manipulations. The current study aimed to assess the growth performance and expression of hepatic growth related genes in the naked neck broiler chicken in response to different dietary energy and protein levels with a hypothesis that high plane of nutrition enhances both of these positively. The results revealed that birds have shown significantly better growth performance under high protein (HP) and high energy (HE) dietary regime. The expression profiles of the genes studied revealed upregulation of IGF-1, IGF-2, and GH under dietary HP and HE regime relative to other protein and energy levels with greater upregulation at 3rd week than the 1st and 5th week of age of birds. The IGFR and GHR mRNA expression was significantly higher under HP and HE dietary regimen with an increasing and decreasing trend from 1st to 5th week of age, respectively. A consistent and significant downregulation of IGFBP-2 was observed under HP and HE regime throughout the feeding trial. The myostatin expression was higher at 3rd week of age followed by 1st week expression. The HP and HE as well as LP (Low protein) and HE diet resulted in significant upregulation of myostatin gene expression in liver. In support to the set hypothesis of this study the high protein and high energy diet resulted in better growth performance of broiler chickens with corresponding upregulation of IGF-1, IGF-2, IGFR, GH, GHR, and Myostatin gene expression and downregulation of IGFBP-2 in liver.

In ovo silver nanoparticle supplementation for improving the post-hatch immunity status of broiler chickens.

Silver nanoparticles (AgNano) are known for their unique physical, chemical and biological properties, enabling cell penetration and anti-inflammatory response. In Experiment 1, the effect of an in ovo administration of AgNano (15 µg/egg; n = 360) at different incubation times (d 7 and d 18) on hatchability parameters was explored. In Experiment 2, post-hatch performance of broilers (42 d, n = 250) was studied after in ovo AgNano administration: Group T1 remained un-injected, Group T2 was the sham control and Groups T3, T4 and T5 were injected with 12.5, 25 and 50 µg AgNano, respectively, at 18 d of incubation. Chick weight, chick to egg weight ratio and hatchability as well average daily gain, average daily feed intake and feed conversion ratio were similar in all treatment groups. No variation was seen in the weight of thymus; however, the bursa and spleen weight was increased (p < 0.05) in Groups T4 and T5 in comparison to Group T1. The in vivo immune response to phytohaemagglutinin-P was increased in Group T3 in comparison to Groups T1 and T2 (p < 0.05), while the response to sheep red blood cells was increased in all AgNano-treated groups in comparison with Group T1 (p < 0.01). The expression of toll-like receptors 2 and 4 genes was up-regulated in AgNano groups in comparison with Groups T1 and T2 (p < 0.01). In summary, an in ovo supplementation of AgNano carried out at d 18 of incubation is effective and modulates the post-hatch immune response without affecting the hatchability, growth and other performance parameters in broilers.

Effects of Ecklonia cava as fucoidan-rich algae on growth performance, nutrient digestibility, intestinal morphology and caecal microflora in weanling pigs.

OBJECTIVE: In the present study, role of increasing levels of Ecklonia cava (seaweed) supplementation in diets was investigated on growth performance, coefficient of total tract apparent digestibility (CTTAD) of nutrients, serum immunoglobulins, cecal microflora and intestinal morphology of weanling pigs. METHODS: A total of 200 weaned pigs (Landrace×Yorkshire×Duroc; initial body weight 7.08±0.15 kg) were randomly allotted to 4 treatments on the basis of body weight. There were 5 replicate pens in each treatment including 10 pigs of each. Treatments were divided by dietary Ecklonia cava supplementation levels (0%, 0.05%, 0.1%, or 0.15%) in growing-finishing diets. There were 2 diet formulation phases throughout the experiment. The pigs were offered the diets ad libitum for the entire period of experiment in meal form. RESULTS: The pigs fed with increasing dietary concentrations of Ecklonia cava had linear increase (p<0.05) in the overall average daily gain, however, there were no significant differences in gain to feed ratio, CTTAD of dry matter and crude protein at both phase I and phase II. Digestibility of gross energy was linearly improved (p<0.05) in phase II. At day 28, pigs fed Ecklonia cava had greater (linear, p<0.05) Lactobacillus spp., fewer Escherichia coli (E. coli) spp. (linear, p<0.05) and a tendency to have fewer cecal Clostridium spp. (p = 0.077). The total anaerobic bacteria were not affected with supplementation of Ecklonia cava in diets. Polynomial contrasts analysis revealed that villus height of the ileum exhibited a linear increase (p<0.05) in response with the increase in the level of dietary Ecklonia cava. However, villus height of duodenum and jejunum, crypt depth, villus height to crypt depth ratio of different segments of the intestine were not affected. CONCLUSION: The results suggest that Ecklonia cava had beneficial effects on the growth performance, cecal microflora, and intestinal morphology of weanling pigs.

In ovo trace element supplementation enhances expression of growth genes in embryo and immune genes in post-hatch broiler chickens.

BACKGROUND: Differential expression of growth- and immunity-related genes and post-hatch performances were evaluated in in ovo zinc (Zn), iodine (I) or selenium (Se) supplemented chicken embryos. RESULT: There was about 9-18% reduction in hatchability of Zn, I or Se supplemented eggs. In ovo trace element supplementation did not improve post-hatch growth. Two-way analysis of data revealed significant effect (P > 0.01) of period, trace elements and their interactions. Expression of hepatic somatotropin, insulin-like growth factor-II and mucin gene was highest at 20(th) embryonic day but decreased during post-hatch periods. In ovo Zn or I supplemented embryos had higher expression of growth-related genes compared to the Se or un-injected control group. Expression of interleukin-6 was higher (P < 0.01) in in ovo I supplemented chicks (2.5-fold) but lower in the Zn and Se groups than in the un-injected control group. However, Zn and Se supplemented chicks had higher cellular immune gene expression. In vivo response to mitogen phytohaemaglutinin was also higher (P < 0.01) in Zn or Se supplemented chicks CONCLUSION: In ovo supplementation of Zn, I and Se did not improve the post-hatch growth, but increased growth-related gene expression. Iodine improved humoral immune gene expression whereas Zn and Se enhanced cell-mediated immune gene expression in broiler chickens. © 2015 Society of Chemical Industry.

In ovo feeding of α-ketoglutaric acid improves hepatic antioxidant-gene expression, plasma antioxidant activities and decreases body temperature without affecting broiler body weight under cyclic heat stress.

The broiler industry is adversely affected by the rise in global temperature. This study investigated the effects of in ovo feeding of α-ketoglutaric acid (AKG) on growth performance, organ weight, plasma metabolite, plasma oxidative stress, rectal temperature (RT), and hepatic mRNA expression of antioxidant-related genes in Arbor Acres broilers subjected to cyclic heat stress (HS). Three hundred fifty fertile eggs during incubation were divided into 5 groups according to AKG concentrations and temperature conditions. After dissolving AKG in distilled water at 0, 0.5, 1.0, and 1.5, 0% AKG was in ovo administered to 2 of the 5 groups whereas the remaining 3 groups received 0.5, 1.0, and 1.5%, respectively. From d 29 to 34 of age, 4 groups of birds received heat stress (HS) at 31°C ± 1°C for 6 h per day while the other group was kept at room temperature (21°C ± 1°C; NT). So, the 5 treatment groups were: 1) 0AKG-NT, where chicks hatched from eggs receiving 0% AKG were reared under thermoneutral conditions. 2) 0AKG-HS, where chicks hatched from eggs receiving 0% AKG were reared under cyclic HS conditions. 3) 0.5AKG-HS, where chicks hatched from eggs receiving 0.5% AKG were reared under cyclic HS conditions. 4) 1.0AKG-HS, where chicks hatched from eggs receiving 1.0% AKG were reared under cyclic HS conditions. 5) 1.5AKG-HS, where chicks hatched from eggs receiving 1.5% AKG were reared under cyclic HS conditions. HS significantly reduced body weight change (ΔBW %) and average daily gain (ADG) without affecting average daily feed intake (ADFI). Feed conversion ratio (FCR) was significantly increased (P = 0.003) in all HS-treated groups. A significant linear decrease in the final RT (P = 0.005) and a change in RT (P = 0.003) were detected with increasing AKG concentration. Total antioxidant capacity (P = 0.029) and antioxidant balance (P = 0.001) in plasma increased linearly with increasing AKG concentration whereas malondialdehyde concentrations were linearly decreased (P = 0.001). Hepatic gene expression of CAT (P = 0.026) and GPX1 (P = 0.001) were dose-dependently upregulated while nicotinamide adenine dinucleotide phosphate oxidase (NOX)1, NOX4, and heat shock protein (HSP)70 were linearly downregulated (P < 0.05). Hence, in ovo injection of AKG was effective in mitigating HS-induced oxidative stress without attenuating the adverse effects on broiler growth.

Embryonic modulation through thermal manipulation and in ovo feeding to develop heat tolerance in chickens.

Healthy chickens are necessary to meet the ever-increasing demand for poultry meat. Birds are subjected to numerous stressful conditions under commercial rearing systems, including variations in the environmental temperature. However, it is difficult to counter the effects of global warming on the livestock industry. High environmental temperature is a stressful condition that has detrimental effects on growth and production performance, resulting in decreased feed intake, retarded growth, compromised gut health, enhanced oxidative stress, and altered immune responses. Traditional approaches include nutritional modification and housing management to mitigate the harmful effects of hot environments. Currently, broiler chickens are more susceptible to heat stress (HS) than layer chickens because of their high muscle mass and metabolic rate. In this review, we explored the possibility of in ovo manipulation to combat HS in broiler chickens. Given their short lifespan from hatching to market age, embryonic life is thought to be one of the critical periods for achieving these objectives. Chicken embryos can be modulated through either temperature treatment or nourishment to improve thermal tolerance during the rearing phase. We first provided a brief overview of the harmful effects of HS on poultry. An in-depth evaluation was then presented for in ovo feeding and thermal manipulation as emerging strategies to combat the negative effects of HS. Finally, we evaluated a combination of the two methods using the available data. Taken together, these investigations suggest that embryonic manipulation has the potential to confer heat resistance in chickens.

Effect of in ovo feeding of γ-aminobutyric acid combined with embryonic thermal manipulation on hatchability, growth, and hepatic gene expression in broilers.

OBJECTIVE: This study investigated the effects of in ovo feeding of γ-aminobutyric acid (GABA) and embryonic thermal manipulation (ETM) on growth performance, organ indices, plasma biochemical parameters, hepatic antioxidant levels, and expression of lipid metabolism-related genes in broilers. METHODS: Two hundred and fifty eggs were assigned to one of four treatments: control eggs incubated under standard conditions (CON); eggs that received an in ovo injection of 10% GABA on day 17.5 of incubation (G10); thermally manipulated eggs between days 10 and 18 of incubation at 39.6°C for 6 h daily (TM); and eggs that received both treatments during incubation (G10+TM). After 28 days of rearing, five birds per treatment were selected for blood and organ sampling. RESULTS: No differences were found in hatchability or growth parameters among different treatment groups. Hepatic gene expression of catalase (CAT) and glutathione peroxidase 1 (GPx1) was upregulated (p = 0.046 and p = 0.006, respectively) in the G10+TM group, while that of nuclear factor erythroid 2-related factor 2 (NRF2) was upregulated (p = 0.039) in the G10 group. In addition, the relative gene expression of NADPH oxidase 1 (NOX1) was significantly lower (p = 0.007) in all treatment groups than that in the CON group. Hepatic fatty acid synthase (FAS) levels and average daily feed intake (ADFI) of last week showed a positive correlation (r = 0.50, p = 0.038). In contrast, the relative gene expression of the extracellular fatty acid-binding protein (EXFAB) and peroxisome proliferator-activated receptor-γ (PPAR-γ) were positively correlated (r = 0.48, p = 0.042 and r = 0.50, p = 0.031) with the overall ADFI of birds. CONCLUSION: Taken together, the results of this study suggest that the combination of in ovo feeding of GABA and ETM can enhance hepatic antioxidant function in broilers.

Dietary supplementation of solubles from shredded, steam-exploded pine particles modifies gut length and cecum microbiota in cyclic heat-stressed broilers.

This study was conducted to investigate the effect of supplementing solubles from steam-exploded pine particles (SSPP) on mitigating the adverse effects of cyclic heat stress (CHS) in broilers which were distributed into 3 dietary treatment groups and 2 temperature conditions. Heat stress (HS) exposure for 6 h daily for 7 d adversely affected performance parameters and rectal temperature of chickens. The absolute and relative weights of the liver and bursa of Fabricius decreased in the CHS group while the relative lengths of the jejunum and ileum increased, which was rescued by dietary supplementation with SSPP. The expression of mucin2 (MUC2) and occludin (OCLN) genes was decreased in CHS birds. The expression of heat shock protein -70 and -90 increased in 0% HS compared to that in 0% NT. Birds supplemented with 0.4% SSPP had higher NADPH oxidase -1 expression than birds in the 0% and 0.1% SSPP treatments. Beta diversity of gut microbiota evaluated through unweighted UniFrac distances was significantly different among treatments. Bacteroidetes was among the 2 most abundant phyla in the cecum, which decreased with 0.1% NT and increased with 0.1% HS in comparison to 0% NT. A total of 13 genera were modified by HS, 5 were altered by dose, and nine showed an interaction effect. In conclusion, CHS adversely affects performance and gut health which can be mitigated with dietary SSPP supplementation that modifies the cecal microbiota in broilers.

A novel gut microbiome-immune axis influencing pathology in HCMV infected infants with neonatal cholestasis.

The interplay of active HCMV infection with gut dysbiosis in the immunopathology of cholestasis in neonates and infants remains unexplored. In this study, we evaluated gut microbiome profiles and immune dysfunction in a cohort of HCMV infected cholestatic infants (IgM positive, N = 21; IgM negative, N = 25) compared to healthy infants, N = 10. HCMV infected IgM positive individuals exhibited increased clinical severity in terms of liver dysfunction, altered CD4+: CD8+ ratio, and elevated Granzyme B levels in cellular immune subsets. Gut microbiome analysis revealed distinct and differential diversity and composition within infected groups aligned with clinical severity reflected through the increased abundance of Gammaproteobacteria, reduced Bifidobacteria, and a unique signature mapping to the HCMV infected IgM negative group. Correlation analyses revealed associations between Bifidobacterium breve, Gammaproteobacteria, Firmicutes, Clostridia, Finegoldia magna, Veillonella dispar, and Granzyme B expressing immune cell subsets. Our study describes a novel gut microbiome-immune axis that may influence disease severity in cholestatic infants with active HCMV infection.

Transient elastography and von Willebrand factor as predictors of portal hypertension and decompensation in children.

Background & Aims: Von Willebrand factor antigen (vWFAg), a protein measured to test the level of vWF released from the vascular endothelium has gained much attention as a marker for portal hypertension (PHT) severity. The objectives of this study were to investigate the use of vWFAg as a biomarker along with liver and spleen stiffness measurements by transient elastography as potential predictors of clinically significant varices (CSV), variceal bleeding (VB) and decompensation in children with PHT. Methods: This observational prospective cohort study included 117 children (median age 10 [IQR 6-14] years) who underwent oesophagogastroduodenoscopy between January'2012 to November'2021 and a validation group of 33 children who underwent the same procedure between December'2021 to March'2023. Measurements of vWFAg and glycoprotein Ib binding activity of VWF (GPIbR) were available in 97 patients in the study group and in all patients in the validation group.Results: vWFAg and GPIbR were significantly higher in children with CSV (223 IU/dl and 166 IU/dl; p = 0.015 and p = 0.04, respectively) and VB (218 IU/dl and 174 IU/dl; p = 0.077 and p = 0.03, respectively) than in those without CSV or VB, respectively. Ninety-six patients had liver and spleen stiffness measurements. Spleen stiffness was significantly higher in patients with CSV compared to those without CSV (p = 0.003). In a chronic liver disease subgroup, a predictive scoring tool based on vWFAg, GPIbR, platelet count, and spleen/liver stiffness measurements could predict CSV with an AUROC of 0.76 (p = 0.04). Conclusions: This study suggests the predictive value of vWF for CSV and VB increases when combined with spleen stiffness, with AUROCs of 0.88 and 0.82, respectively. Hence, a combination of biomarkers could assist clinicians in diagnosing CSV, preventing unnecessary invasive procedures. Impacts and implications: Surveillance endoscopies in children with portal hypertension (PHT) have their own risks and non-invasive markers, such as von Willebrand factor antigen, glycoprotein Ib binding activity of VWF (GPIbR), and transient elastography could be used to predict clinically significant varices, variceal bleeding and disease compensation in children with PHT. Such non-invasive markers for PHT and varices are lacking in the paediatric population. The results show that von Willebrand factor and GPIbR along with transient elastography can be used to formulate a scoring system which can be used as a clinical tool by paediatric hepatologists to monitor the progression of PHT and risk of bleeding, and hence to stratify the performance of invasive endoscopic procedures under general anaesthesia. However, there is a need to validate the scoring system in children with portal vein thrombosis and for hepatic decompensation in a multi-centre registry in the future.

Dietary supplementation of solubles from shredded, steam-exploded pine particles modulates cecal microbiome composition in broiler chickens.

This study evaluated the effects of supplementing solubles from shredded, steam-exploded pine particles (SSPP) on growth performances, plasma biochemicals, and microbial composition in broilers. The birds were reared for 28 days and fed basal diets with or without the inclusion of SSPP from 8 days old. There were a total of three dietary treatments supplemented with 0% (0% SSPP), 0.1% (0.1% SSPP) and 0.4% (0.4% SSPP) SSPP in basal diets. Supplementation of SSPP did not significantly affect growth or plasma biochemicals, but there was a clear indication of diet-induced microbial shifts. Beta-diversity analysis revealed SSPP supplementation-related clustering (ANOSIM: r = 0.31, p < 0.01), with an overall lower (PERMDISP: p < 0.05) individual dispersion in comparison to the control group. In addition, the proportions of the Bacteroides were increased, and the relative abundances of the families Vallitaleaceae, Defluviitaleaceae, Clostridiaceae, and the genera Butyricicoccus and Anaerofilum (p < 0.05) were significantly higher in the 0.4% SSPP group than in the control group. Furthermore, the linear discriminant analysis effect size (LEfSe) also showed that beneficial bacteria such as Ruminococcus albus and Butyricicoccus pullicaecorum were identified as microbial biomarkers of dietary SSPP inclusion (p < 0.05; | LDA effect size | > 2.0). Finally, network analysis showed that strong positive correlations were established among microbial species belonging to the class Clostridia, whereas Erysipelotrichia and Bacteroidia were mostly negatively correlated with Clostridia. Taken together, the results suggested that SSPP supplementation modulates the cecal microbial composition of broilers toward a "healthier" profile.

Microarray analysis and PCR validation of genes associated with facultative parthenogenesis in Meleagris gallopavo (Turkey).

A cDNA microarray containing 43,661 differentially expressed genes was carried out on the blastoderm of fertilized and facultative parthenogenic turkey embryos at different hours of development. The total number of up-regulated (UR) and down-regulated (DR) genes at 0, 12, and 24 h of development were 725 and 1436, 942 and 599, and 589 and 1044, respectively. Common genes between 0 and 12 h, 12 and 24 h, and 0 and 24 h were 55, 67, and 110, respectively. The proportion of genes showing above 50-fold UR and DR at 0, 12, and 24 h of development were 2.0% and 1.5%, 0.5% and 1.2%, and 0.2% and 1.1%, respectively. Eight UR genes were validated (APOA1, THRAP3, ARL14EP, PSAP, MOG, MYBPC2, MTIF3 and EDG4) and relative expression of six of them was significantly higher (P ≤ 0.05) in parthenogenic embryos, while two genes showed non-significant (P ≥ 0.05) variation. The expression of BCL11A, PRP4B, TCP1, and TPI1 genes was significantly (P ≤ 0.05) DR in parthenotes in the micro-array study, while the TCP1 gene was up-regulated, and there was no variation in TPI1 gene expression in the PCR validation study. In conclusion, our findings demonstrate differential expression of a large number of genes in parthenotes at different stages of embryo development compared to fertilized embryos. Up-regulation of APOA1, MYBPC2, TCP1, and THRAP3 genes, suggest their crucial role in spontaneous facultative parthenogenic development in turkey birds.

Embryonic manipulations modulate differential expressions of heat shock protein, fatty acid metabolism, and antioxidant-related genes in the liver of heat-stressed broilers.

In this study, the effects of in ovo feeding of γ-aminobutyric acid (GABA) and embryonic thermal manipulation (TM) on plasma biochemical parameters, organ weights, and hepatic gene expression in broilers exposed to cyclic heat stress (32 ± 1°C for 8 days) (HS) were investigated. A total of 175 chicks were assigned to five treatments: chicks hatched from control eggs (CON); chicks hatched from control eggs but exposed to HS (CON+HS); chicks hatched from eggs injected at 17.5 days of incubation with 0.6mL of 10% GABA and exposed to HS (G10+HS); chicks hatched from thermally manipulated eggs (39.6°C, 6h/d from embryonic days 10 to 18) and exposed to HS (TM+HS); chicks hatched from eggs that received both previous treatments during incubation and exposed to HS (G10+TM+HS). Results revealed that on day 36 post-hatch, hepatic NADPH oxidase 1 (P = 0.034) and 4 (P = 0.021) genes were downregulated in the TM+HS and G10+TM+HS compared to the CON+HS group. In addition, while acetyl-CoA carboxylase gene expression was reduced (P = 0.002) in the G10+TM group, gene expression of extracellular fatty acid-binding protein and peroxisome proliferator-activated receptor-γ was lower (P = 0.045) in the TM+HS group than in the CON+HS group. HS led to higher gene expression of heat shock protein 70 (HSP70) and 90 (HSP90) (P = 0.005, and P = 0.022). On the other hand, the TM+HS group exhibited lower expression of both HSP70 (P = 0.031) and HSP90 (P = 0.043) whereas the G10+TM+HS group had a reduced (P = 0.016) HSP90 expression compared to the CON+HS. MANOVA on different gene sets highlighted an overall lower (P = 0.034) oxidative stress and lower (P = 0.035) heat shock protein expression in the G10+TM+HS group compared to the CON+HS group. Taken together, the current results suggest that the combination of in ovo feeding of GABA with TM can modulate HSPs and antioxidant-related gene expression in heat-stressed broilers.

Impact of embryonic manipulations on core body temperature dynamics and survival in broilers exposed to cyclic heat stress.

Ambient temperature-associated stress has been shown to affect the normal physiological functions of birds. The recent literature indicated that both, embryonic thermal manipulation (ETM) and in ovo feeding (IOF) of γ-aminobutyric acid (GABA) can mitigate the deleterious effects of heat stress (HS) in young broiler chicks. Therefore, this study intended to assess the effects of cyclic HS (32 ± 1 °C, 4 h/day from day 29 to 35) on rectal temperatures (RTs) and survival in broiler chickens after ETM and in IOF of GABA. A total of 275 RT data points and survival data were collected from chicks assigned to the following five treatments: chicks hatched from control eggs (CON); chicks hatched from control eggs but exposed to HS (CON + HS); chicks hatched from eggs injected at 17.5 days of incubation with 0.6 mL of 10% GABA and exposed to HS (G10 + HS); chicks hatched from thermally manipulated eggs (39.6 °C, 6 h/day from embryonic days 10 to 18) and exposed to HS (TM + HS); chicks hatched from eggs that received both previous treatments during incubation and exposed to HS (G10 + TM + HS). Under thermoneutral conditions, RTs increased quadratically from 39.9 ± 0.2 °C at hatching to 41.4 ± 0.1 °C at 8 days of age. When exposed to cyclic HS during the last week of rearing, the birds' RTs tended to decrease at the end of the heat stress challenge (from 43.0 ± 0.2 °C on day 29 to 42.4 ± 0.1 °C on day 35). A stepwise Cox regression indicated that treatment was predictive of birds' survival. Hazard ratios (HR) and their confidence intervals (CI) were calculated to assess the likelihood of death during the trial. The birds, belonging to the G10 + TM + HS group, were less likely to die under HS (HR 0.11, 95% CI 0.02 to 0.91, P = 0.041) compared to the CON + HS birds. Taken together, the combination of ETM and GABA IOF may help mitigate the drawbacks of cyclic HS by improving the survival of broilers.

Effects of In Ovo Injection of α-Ketoglutaric Acid on Hatchability, Growth, Plasma Metabolites, and Antioxidant Status of Broilers.

Recently, α-ketoglutaric acid (AKG) has gained importance as an antioxidant. Its dietary supplementation in animals and humans has proved beneficial. Moreover, an extensive group of studies on in ovo feeding has proved that it produces better day-old chicks and overall performance. Combining the two, we hypothesized that in ovo feeding of AKG could improve the antioxidant status in addition to chick quality and broiler performance. At 17.5 days of incubation, eggs were divided into one of five groups: eggs that received (i) no injection (U-CON), (ii) distilled water (DDW) only (0 AKG), (iii) 0.5% AKG dissolved in DDW (0.5 AKG), (iv) 1.0% AKG dissolved in DDW (1.0 AKG), or (v) 1.5% AKG dissolved in DDW (1.5 AKG). Chicks were raised until 21 days of age. Biological samples were collected on day 0 and day 21. Body weight (p = 0.020), average daily gain (p = 0.025), and average daily feed intake (p = 0.036) were found to quadratically increase with the amount of AKG during the grower phase. At day 0, the absolute (p = 0.040) and relative weight (p = 0.035) of the liver increased linearly with an increasing amount of AKG. The 0.5 AKG group had significantly higher plasma protein (p = 0.025), absolute and relative heart indices at day 0 (p = 0.006). An in ovo feeding of AKG improved the plasma antioxidant capacity of chicks at day 0 as compared to 0 AKG. AKG effect was seen on the plasma antioxidant balance, which increased linearly with the increasing dose of in ovo AKG. Furthermore, 1.0 AKG and 1.5 AKG showed a significant (p = 0.002) upregulation of the hepatic mRNA expression of nuclear factor erythroid 2-related factor (NRF2) in comparison to 0 AKG. The results imply that without negatively affecting hatchability performance, in ovo feeding of AKG has beneficial effects on the antioxidant status of broilers.

Effects of Dietary Supplementation of Solubles from Shredded, Steam-Exploded Pine Particles on the Performance and Cecum Microbiota of Acute Heat-Stressed Broilers.

Heat stress (HS) negatively influences livestock productivity, but it can be, at least in part, mitigated by nutritional interventions. One such intervention is to use byproducts from various sources that are likely to be included in the consumer chain. Thus, the present study investigated the effects of dietary supplementation of solubles from shredded, steam-exploded pine particles (SSPPs) on the performance and cecum microbiota in broilers subjected to acute HS. One-week-old Ross 308 broilers (n = 108) were fed 0%, 0.1%, or 0.4% SSPP in their diets. On the 37th day, forty birds were allocated to one of four groups; namely, a group fed a control diet without SSPPs at thermoneutral temperature (NT) (0% NT) and acute heat-stressed birds with 0% (0% HS), 0.1% (0.1% HS), and 0.4% (0.4% HS) SSPP-supplemented diets. The NT was maintained at 21.0 °C, while the HS room was increased to 31 °C. The final BW, percent difference in body weight (PDBW), and feed intake (FI) were lower in HS birds, but PDBW was reversely associated with dietary SSPP. Similarly, HS birds had a higher rectal temperature (RT) and ΔT in comparison to birds kept at NT. The FI of SSPP-supplemented birds was not significant, indicating lower HS effects. Plasma triglyceride was decreased in HS birds but not affected in 0.1% HS birds in comparison to 0% NT birds. OTUs and Chao1 were increased by 0.1% HS compared to 0% NT. Unweighted Unifrac distances for 0.1% HS were different from 0% NT and 0.4% HS. The favorable bacterial phylum (Tenericutes) and genera (Faecalibacterium and Anaerofustis) were increased, while the pathogenic genus (Enterococcus) was decreased, in SSPP-supplemented birds. In sum, production performances are negatively affected under acute HS. Dietary supplementation of SSPPs is beneficial for improving community richness indices and unweighted Unifrac distances, and it enhanced the advantageous bacterial phyla and reduced virulent genera and triglyceride hydrolysis in acute HS broilers. Our results indicate that dietary SSPPs modulates the microbial profile of the cecum while resulting in relatively less weight loss and lower rectal temperature compared to control.