In ovo feeding of L-arginine and selenium nanoparticles influences post-hatch growth, muscle development, antioxidant status, and meat quality in slow-growing chickens.

This study investigated the effects of in ovo feeding (IOF) of L-arginine (L-Arg), selenium nanoparticles (SeNP), and a combination of L-Arg and SeNP on the hatchability, post-hatch growth, muscle development, antioxidant status, and meat quality of slow-growing chickens. On day 18 of incubation, a total of 960 fertilized eggs with similar weights were randomly assigned to four treatment groups with four replicates of 60 eggs each: 1) non-injected control group (Control); injected with 1% of L-Arg (IOF_L-Arg); 3) injected with 0.3 µg/egg of SeNP (IOF_SeNP); and 4), injected with 1% of L-Arg and 0.3 µg/egg of SeNP (IOF_L-Arg+SeNP). A completely randomized design was used. After hatching, 640 mixed-sex chicks were allocated to four treatment groups and split into four replicate pens (40 birds per pen). All groups of chicks were fed with commercial feed ad libitum until they reached 63 days of age and were subsequently weighed and slaughtered. The results of the present study showed that hatchability was similar among treatments. Final BW or breast muscle yield were not affected (P > 0.05) by IOF treatment. Chickens treated with IOF_L-Arg+SeNP exhibited decreased feed conversion ratio, drip loss, and increased protein content in breast meat (P < 0.05). The IOF_L-Arg+SeNP group exhibited a higher density of breast muscle fibers than the control group (P < 0.05). Overall, in ovo feeding of L-Arg combined with SeNP resulted in improved feed efficiency and enhanced antioxidant capacity at hatch without any adverse effects on chicken hatchability, health, or subsequent growth. Furthermore, meat from chickens in the IOF_L-Arg+SeNP group exhibited a preferable texture with a higher protein content.

Supplemental L-arginine promotes hepatocyte proliferation and alters liver fatty acid metabolism in the late embryonic phase: an RNA-seq analysis.

The in ovo feeding (IOF) of L-arginine (L-Arg) to chick embryos is a viable method for improving early intestinal development, subsequently leading to an acceleration in growth rate during the posthatch stage. However, the liver, being the pivotal organ for energy metabolism in poultry, the precise effects and mechanisms of L-Arg on the liver development and metabolism remain unclear. To elucidate these, the present study injected 2 doses of L-Arg (10 mg/egg and 15 mg/egg) into the embryos of Hongyao chickens at 17.5 d of incubation, subsequently incubating them until d 19 for further analysis. IOF of 15 mg L-Arg/egg significantly increased the organ indices of liver and small intestine, as well as the duodenal villus height/crypt depth. RNA-Seq analysis of liver tissues showed that the metabolism of xenobiotics, amino acid metabolism, and the fatty acid metabolism were significantly enriched in L-Arg injection group. The core differentially expressed genes (DEGs) were primarily involved in cell proliferation and fatty acid metabolism. The CCK8 assays revealed that supplemental L-Arg significantly enhanced the proliferation of primary embryo hepatocytes and leghorn male hepatoma (LMH) cells. Upregulation of core DEGs, including HBEGF, HES4, NEK3, EGR1, and USP2, significantly promoted the proliferation of liver cells. Additionally, analysis of triglyceride and total cholesterol content, as well as oil red O staining, indicated that supplemental L-Arg effectively reduced lipid accumulation. Overall, L-Arg supplementation in late chick embryos may promote early liver and small intestine development by reducing liver lipid deposition and enhancing energy efficiency, necessitating further experimental validation. This study provides profound insights into the molecular regulatory network of L-Arg in promoting the development of chicken embryos. The identified DEGs that promote cell proliferation and lipid metabolism can serve as novel targets for further developing methods to enhance early development of chicken embryos.

Effects of in ovo supplementation of selenium (Se) and zinc (zn) on hatchability and production performance of broiler chickens.

The current research was conducted to assess the effect of in ovo feeding (IOF) of selenium (Se) and zinc (Zn) on hatchability, production performance, liver, intestinal morphology, antioxidant levels and expression levels of immune-related genes in broiler chickens. A total of 400 fertilized eggs were equally divided into four groups: control (non-injected), sham (in ovo injection of 0.75% NaCl), Se (@ 1.5 µg/egg in ovo injection) and Zn (500 µg/egg in ovo injection) groups respectively. On the seventeenth day of incubation, treatment solutions were administered into amniotic fluid of fertilized eggs. The results revealed that Se and Zn supplementation significantly (P < 0.05) enhanced hatchability, post-hatch growth, organ development, and liver antioxidant capability. Histopathological examination revealed a typical hepatocyte morphology, well-arranged cells, and a significant (P < 0.05) decrease in apoptosis in both selenium and zinc groups. Additionally, selenium and zinc produced auspicious effects on intestinal epithelium and villi surface area. Interestingly, our results revealed that IOF of Se and Zn modulated the expression of immune-related genes in comparison to the control and sham groups. Conclusively, IOF of Se and Zn augmented health and productivity by enhancing the cellular immunity in the broiler chickens, thus IOF can be utilized as an effective strategy to promote health and immunity in broiler chickens.

In ovo injection of bee pollen extract on hatchability, chick quality, glycogen reserves and production performance in broiler chickens.

This study aimed to investigate the effects of in ovo injection of bee pollen (BP) extract on some hatching traits, glycogen reserves and production performance in broilers. A total of 886 eggs was randomly assigned to 5 treatments: the 0.9% NaCl diluent-injected control group, and the groups that were injected with BP extract at 3 different concentrations: BP-1.5%, BP-2.5% and BP-5.0% (7, 12 and 25 mg/egg, respectively). The last group received a carbohydrate solution (CS). At 18 d of incubation, 500 µL of each solution was injected into the air sac of each egg of the injected groups. After hatching, a total of 570 one-day-old chicks were distributed into 5 groups (in ovo injection) with 3 replicates with 38 birds. The in ovo injection of BP at a dose of 5% and CS resulted in lower hatching rates (P < 0.05) as compared with the control group. The level of glycogen in the muscle tissue of newly hatched birds was not significantly different (P > 0.05). Supplementation of embryos with BP extract also caused a significant increase in the length of chicks in the BP-1.5% group (P < 0.001). Hatched chicks from the injected eggs had a lower liver glycogen content than that of the control. Based on the results, it was concluded that high-quality day-old chicks could be obtained by in ovo injection. Supplementation of chicks with 1.5% BP extract had a beneficial effect on BW during the first rearing period and showed a hypocholesterolemic effect in young chicks. At the same time, an adverse effect of a high dose of BP (5%) and a carbohydrate solution administered in ovo on the hatchability level of chicks was shown. The implementation of the carbohydrate mixture resulted in a deterioration of biochemical indices in the plasma of newly hatched chicks, especially in the case of transaminase activity.

The potential of in ovo-fed amino acids to alleviate the effects of heat stress on broiler chickens: effect on performance, body temperature, and oxidative status during the finisher phase.

The aim of the current study was to investigate the potential of in ovo-fed amino acids (AA) to reduce the effects of heat stress on finishing broiler chickens. To achieve this, a total of 1,400 fertile hatching eggs were randomly distributed into 5 groups (n = 280/group) and injected with one of the following in ovo treatments on embryonic day 18: 52 µL of sterile diluent/egg (CTRL), CTRL + 1.0 mg of L-Leucine (T1), CTRL + 0.45 mg of leucine + 1.15 mg of methionine (T2), CTRL + 3.0 mg of methionine + 2.0 mg of cysteine (T3), and CTRL + 0.40 mg of leucine + 1.60 mg of methionine + 1.60 mg of cysteine (T4). After hatch, chicks were allocated according to a complete randomized block design comprising 2 thermal conditions: thermoneutral (24°C, 45% RH) and heat stress (34°C, 55-60% RH) with 5 pens/group/condition. The cyclical heat stress regimen (10 h/d) was then applied from d 29 to d 34. Compared to the CTRL group, T3 and T4 exhibited a higher BW during the starter phase (P < 0.001). T4 also had a lower feed conversion ratio (FCR) than CTRL during this same phase (P = 0.03). During the grower phase, males of all treatment groups consistently exhibited higher BW compared to the CTRL group, which was not observed among female birds (PSex × TRT = 0.005). During the finisher phase, the in ovo treatment effect on performance was not significant. However, heat-stressed birds from treatment group T3 and T4 exhibited lower facial temperatures (Pday × TRT < 0.001) as well as lower plasma (Pcondition x TRT = 0.039) and liver (Pcondition x TRT < 0.001) malonaldehyde concentrations compared to the CTRL group. In conclusion, in ovo-fed AA have the potential to modulate the effects of heat stress on finishing broiler chickens by limiting its detrimental consequences, including increased body temperature and oxidative damage.

Impact of in ovo feeding of grape pomace extract on the growth performance, antioxidant status, and immune response of hatched broilers.

Delivering natural antioxidants via in ovo feeding holds promise for enhancing the antioxidant status and performance of chickens. Therefore, The objective of this study was to evaluate the impacts of in ovo feeding during early embryonic development using grape pomace extract as a natural antioxidant on hatchability, productive performance, immune response, and antioxidant status in broilers. A total of 900 fertile broiler eggs from the Arbor Acres strain were utilized. Each egg was individually weighed, with egg weights ranging from 61.88 ± 3 g. On the 17.5th d of incubation (DOI), the fertile eggs were divided into 6 groups. The first treatment group was untreated and designated as the control (C). The second group was the sham group (Sh), receiving a simulated injection. The third group, designated as the vehicle group (V), was injected with 100 µl of dimethyl sulfoxide (DMSO). The fourth group received an injection of 100 µL of grape pomace dissolved in DMSO at a concentration of 2 mg (T2). Similarly, the fifth and sixth groups were injected with 100 µL of grape pomace dissolved in DMSO at concentrations of 4 mg and 6 mg, (T4), (T6) respectively. Subsequently, all groups were raised under uniform conditions in terms of management, environment, and nutrition till 5 wk of age. The grape pomace extract (GPE), obtained is rich in total phenolic content (16.07 mg/g), total flavonoid content (7.42 mg/g), and total anthocyanin (8.37 mg/g). Grape pomace extract has exhibited significant antioxidant properties as evidenced by its effectiveness in DPPH scavenging and reducing power assays. Significant improvements in body weight at hatch were observed with in ovo feeding of grape pomace extract, particularly at the 4 mg level, surpassing the effectiveness of the 2 mg and 6 mg grape pomace levels, and this enhancement in body weight continued until the age of 5 wk. GPE injection also led to a significant reduction in cholesterol levels, with the lowest levels recorded for the T4 group. Plasma total Antioxidant Capacity (TAC) levels were significantly elevated in groups treated with T4, T6, and T2 compared to the control group. Conversely, the control group showed a significant increase (P < 0.01) in plasma malondialdehyde (MDA) levels. The immune response of hatched chicks from grape pomace extract-injected groups, especially the T4 group, exhibited improvement through increased IgM and IgG. These findings demonstrate that in ovo feeding of GPE, particularly at a dosage of 4 mg, enhances growth performance, immune response, and antioxidant status in hatched chicks. Thus, administering natural antioxidants, such as grape pomace extract, to developing broiler embryos via in ovo feeding could serve as a valuable strategy for enhancing the subsequent post-hatch productive performance, as well as bolstering the antioxidant and immunological status of broiler chicks.

Short Communication: The effect of in ovo creatine monohydrate on early pen use activity in chicks.

First-week survival and egg hatchability are lower in chicks from younger broiler breeder hen flocks. Creatine is a naturally occurring compound synthesised from the amino acid arginine or obtained from the diet and is important in the storage and transport of energy. Previous research found an improvement in the hatch rate but no posthatch performance improvements when fertile eggs from young breeder hens were injected with creatine monohydrate (CrM) on embryonic day 14. This pilot study aimed to further investigate the possibility of early posthatch improvements by examining the activity of chicks during the 1st week posthatch. Behaviours were broadly classified as active or inactive, the pen was split into three areas, and the amount of time spent in the heat lamp, feed hopper, or drinker line areas was recorded. Chicks given in ovo CrM spent less time in the heat lamp area over the whole 7 days compared to saline (t = 2.352, P = 0.021) and control groups (t = 3.336, P = 0.003) and more time in the feed hopper area during the first 4 days compared to the control group (t = 2.174, P = 0.033). This finding suggests that creatine may improve energy reserves in young chicks allowing them to spend more time away from the heat lamp.

N-Carbamylglutamate in ovo feeding improves carcass yield, muscle fiber development, and meat quality in broiler chickens.

BACKGROUND: Insufficient endogenous nutrients in the broiler embryo can lead to muscle gluconeogenesis, which ultimately affects the post-hatching performance of chicks. This study investigated the effects of in ovo feeding (IOF) of N-carbamylglutamate (NCG) on the growth hormones, carcass yield, and meat quality in broilers. Fertile eggs from a 30-week-old Ross 308 breeder flock were divided into three treatment groups: NC (non-injection), SC (100 µL saline-injection), and NCG (2 mg NCG injection). Each group had six replicates, with 70 eggs per replicate during incubation. Injections were administered on the 17.5th day of embryonic development. After hatching, 270 chicks were selected for 42-day rearing for further sampling. RESULTS: Chicks in the NCG group had significantly higher body weight (BW) and average daily gain (ADG) at the growing phase, increased growth and testosterone hormone in both feeding phases (21 and 42 days), and improved average daily gain (ADG) and food conversion ratio (FCR) in both grower and entire feeding phases (P < 0.05). Triiodothyronine (T3) and tetraiodothyronine (T4) levels, carcass yield, dressing, drum weight, breast muscle weight, drumstick weights, thighs, pectoralis major, and their part percentage of carcass were improved in the NCG group (P < 0.05), these effects were varied along feeding phases. Moreover, IOF of the NCG also improved pectoralis breast muscle color values at 24 h post mortem (P < 0.05). CONCLUSION: These results suggest that NCG injection at the late embryonic age of broiler enhances growth performance and meat quality throughout the lifespan and this can probably be attributed to an increase in thyroid and testosterone hormones, indicating potential involvement in metabolic and nutrient partitioning pathway regulation. © 2024 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.

Zinc glycinate alleviates LPS-induced inflammation and intestinal barrier disruption in chicken embryos by regulating zinc homeostasis and TLR4/NF-κB pathway.

The effect of an immune challenge induced by a lipopolysaccharide (LPS) exposure on systemic zinc homeostasis and the modulation of zinc glycinate (Zn-Gly) was investigated using a chicken embryo model. 160 Arbor Acres broiler fertilized eggs were randomly divided into 4 groups: CON (control group, injected with saline), LPS (LPS group, injected with 32 µg of LPS saline solution), Zn-Gly (zinc glycinate group, injected with 80 µg of zinc glycinate saline solution) and Zn-Gly+LPS (zinc glycinate and LPS group, injected with the same content of zinc glycinate and LPS saline solution). Each treatment consisted of eight replicates of five eggs each. An in ovo feeding procedure was performed at 17.5 embryonic day and samples were collected after 12 hours. The results showed that Zn-Gly attenuated the effects of LPS challenge-induced upregulation of pro-inflammatory factor interleukin 1ß (IL-1ß) level (P =0.003). The LPS challenge mediated zinc transporter proteins and metallothionein (MT) to regulate systemic zinc homeostasis, with increased expression of the jejunum zinc export gene zinc transporter protein 1 (ZnT-1) and elevated expression of the import genes divalent metal transporter 1 (DMT1), Zrt- and Irt-like protein 3 (Zip3), Zip8 and Zip14 (P < 0.05). A similar trend could be observed for the zinc transporter genes in the liver, which for ZnT-1 mitigated by Zn-Gly supplementation (P =0.01). Liver MT gene expression was downregulated in response to the LPS challenge (P =0.004). These alterations caused by LPS resulted in decreased serum and liver zinc levels and increased small intestinal, muscle and tibial zinc levels. Zn-Gly reversed the elevated expression of the liver zinc finger protein A20 induced by the LPS challenge (P =0.025), while Zn-Gly reduced the gene expression of the pro-inflammatory factors IL-1ß and IL-6, decreased toll-like receptor 4 (TLR4) and nuclear factor kappa-B p65 (NF-κB p65) (P < 0.05). Zn-Gly also alleviated the LPS-induced downregulation of the intestinal barrier gene Claudin-1. Thus, LPS exposure prompted the mobilization of zinc transporter proteins and MT to perform the remodeling of systemic zinc homeostasis, Zn-Gly participated in the regulation of zinc homeostasis and inhibited the production of pro-inflammatory factors through the TLR4/NF-κB pathway, attenuating the inflammatory response and intestinal barrier damage caused by an immune challenge.

Effects of exposing Japanese quail eggs to a low dose of gamma radiation and in ovo feeding by two sources of trace elements on embryonic development activities.

The present study investigated the influence of exposing quail eggs to low-dose gamma radiation (GR) and in ovo feeding with 2 sources of a mixture of trace elements (Zn, Fe, and Cu), including sulfate (TES) and loaded with montmorillonite (TEM), on embryonic development activities and prehatch quality. A total of 960 eggs on the seventh day of incubation were randomly divided into 6 groups (160 eggs/group) with 4 replicate of 40 eggs in each. A 3 × 2 factorial arrangement experiment was performed and included 3 sources in ovo feeding with a mixture of trace elements (Zn, Fe, and Cu), including 0 mg/egg, 50 mg TES/egg, and 50 mg TEM/egg with egg irradiation using 0 and 0.2 Gy from GR. Eggs injected with 50 mg TEM/egg and exposed to 0.2 Gy from GR (TEM/GR) was significantly (P ≤ 0.05 and 0.01) higher in hatchability, hatch body weight, and relative organ weight (liver, gizzard, proventriculus, heart, and intestine). The obtained results indicated significant (P ≤ 0.05) decreased in the serum concentration of malondialdehyde (MDA) in TEM/GR group. There was significant (P ≤ 0.05) increased of catalase (CAT) activity and the concentrations of growth hormone (GH) and insulin-like growth factor-1 (IGF-1) in TEM/GR group; however; total antioxidant capacity (T-AOC) was significant (P ≤ 0.05) increased in CT/GR group. Serum concentrations of immunoglobulin M (IgM) (P ≤ 0.05) and tumor necrosis factor-alpha (TNF-α) were increased in the TEM/CR group; the concentration of transforming growth factor beta (TGF-ß) significant (P ≤ 0.05) increased in the TEM/GR group; and interleukins (IL6 and IL10) showed no significant differences among the groups. Our results showed increase in thyroxine and myostatin concentrations with TES/CR and CT/GR of our study groups, respectively. The relative mRNA expression levels of the GH, IGF-1, and Fas cell surface death receptor (FAS) genes were significantly (P ≤ 0.05 and 0.01) upregulated in the liver tissue of the TEM/GR group compared with the other groups. In conclusion, TEM/GR was the best treatment for improving prehatch quality, increasing serum antioxidant enzyme activities, and promoting the expression of growth and immune genes in fertilized quail eggs.

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.

In-ovo feeding with creatine monohydrate: implications for chicken energy reserves and breast muscle development during the pre-post hatching period.

The most dynamic period throughout the lifespan of broiler chickens is the pre-post-hatching period, entailing profound effects on their energy status, survival rate, body weight, and muscle growth. Given the significance of this pivotal period, we evaluated the effect of in-ovo feeding (IOF) with creatine monohydrate on late-term embryos' and hatchlings' energy reserves and post-hatch breast muscle development. The results demonstrate that IOF with creatine elevates the levels of high-energy-value molecules (creatine and glycogen) in the liver, breast muscle and yolk sac tissues 48 h post IOF, on embryonic day 19 (p < 0.03). Despite this evidence, using a novel automated image analysis tool on day 14 post-hatch, we found a significantly higher number of myofibers with lower diameter and area in the IOF creatine group compared to the control and IOF NaCl groups (p < 0.004). Gene expression analysis, at hatch, revealed that IOF creatine group had significantly higher expression levels of myogenin (MYOG) and insulin-like growth factor 1 (IGF1), related to differentiation of myogenic cells (p < 0.01), and lower expression of myogenic differentiation protein 1 (MyoD), related to their proliferation (p < 0.04). These results imply a possible effect of IOF with creatine on breast muscle development through differential expression of genes involved in myogenic proliferation and differentiation. The findings provide valuable insights into the potential of pre-hatch enrichment with creatine in modulating post-hatch muscle growth and development.

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.

The Effects of In Ovo Feeding of Selenized Glucose on Selenium Concentration and Antioxidant Capacity of Breast Muscle in Neonatal Broilers.

This study aims to investigate the impacts of in ovo feeding (IOF) of selenized glucose (SeGlu) on selenium (Se) level and antioxidant capacity of breast muscle in newborn broilers. After candling on 16 day of incubation, a total of 450 eggs were randomly divided into three treatments. On the 17.5th day of incubation, eggs in a control treatment were injected with 0.1 mL of physiological saline (0.75%), while the 2nd group and 3rd group were supplied with 0.1 mL of physiological saline containing 10 µg Se from SeGlu (SeGlu10 group) and 20 µg Se from SeGlu (SeGlu20 group). The results showed that in ovo injection in both SeGlu10 and SeGlu20 increased the Se level and reduced glutathione concentration (GSH) in pectoral muscle of hatchlings (P < 0.05). Compared with the control group, the SeGlu20-treated chicks significantly enhanced the activity of the superoxide dismutase (SOD) and mRNA expression of NAD(P)H quinone dehydrogenase 1 (NQO1) in breast muscle, while there was upregulation in mRNA expressions of glutathione peroxidase 1 (GPX-1) and thioredoxin reductase 1 (TrxR1) and higher total antioxidant capacity (T-AOC) in SeGlu10 treatment (P < 0.05). However, no significant difference on enzyme activities of glutathione peroxidase (GR), glutathione reductase, thioredoxin reductase, concentration of malondialdehyde, and free radical scavenging ability (FRSA) of superoxide radical (O2-•) and hydroxyl radical (OH•) was observed among the three treatments (P > 0.05). Therefore, IOF of SeGlu enhanced Se deposition in breast muscle of neonatal broilers. In addition, in ovo injection of SeGlu could increase the antioxidant capacity of newborn chicks possibly through upregulating the mRNA expression of GPX1, TrxR1, and NQO1, as well as the SOD activity.

Effects of in ovo feeding of chlorogenic acid on antioxidant capacity of postnatal broilers.

In this study, chlorogenic acid (CGA) was injected into the amniotic cavity of chicken embryos to study the effects of in ovo feeding of CGA on the antioxidant capacity of postnatal broilers. On the 17th day of embryonic age, a total of 300 healthy broiler fertile eggs with similar weights were randomly subjected to five groups as follows; in ovo injection with 0.5 ml CGA at 4 mg/egg (4CGA) or 7 mg/egg (7CGA) or 10 mg/egg (10CGA), or sham-injection with saline (positive control, PC) or no injection (negative control, NC). Each group had six replicates of ten embryos. Six healthy chicks with similar body weights hatched from each replicate were selected and reared until heat stress treatment (35°C ± 1°C, 8 h/d) at 28-42 days of age. The results showed that there was no significant difference in the hatching rate between the groups (p > 0.05). After heat stress treatment, 4CGA group showed an improved intestinal morphology which was demonstrated by a higher villus height in the duodenum and a higher villus height/crypt depth ratio in the jejunum, compared with the NC group (p < 0.05). The antioxidant capacity of chickens was improved by in ovo feeding of CGA since 4CGA decreased the plasma content of malondialdehyde (MDA) (p < 0.05), whereas, it increased the superoxide dismutase (SOD), glutathione peroxidase (GPX), and catalase (CAT) activities compared with NC group (p < 0.05). Also, the MDA content of the different injection groups had a quadratic effect, with the 4CGA group having the lowest MDA content (P quadratic < 0.05). In the duodenum, 4CGA injection significantly increased the mRNA expressions of nuclear factor erythroid 2-related factor 2 (Nrf2), heme oxygenase 1 (H O -1), glutathione synthetase (GSS), and SOD1 compared to the NC and PC groups (p < 0.05). The mRNA expressions of glutathione reductase (GSR) and GPX7 were significantly increased in all CGA-treated groups compared with the PC group (p < 0.05), while the mRNA expression of CAT was significantly increased by 4CGA group than the NC group (p < 0.05). The mRNA expressions of epigenetic-related genes, ten eleven translocation 1 and 2 (Tet1 and Tet2), and DNA-methyltransferase 3 alpha (DNMT3A) in the duodenum of 4CGA injected group was significantly increased compared with the NC and PC groups (p < 0.05). The mRNA expressions of Nrf2, SOD1, and Tet2 showed a significant quadratic effects with the 4CGA group having the highest expression (P quadratic < 0.05). In conclusion, in ovo feeding of CGA alleviated heat stress-induced intestinal oxidative damage. Injection with CGA of 4 mg/egg is considered most effective due to its actions in improving intestinal antioxidant capacity, especially in the duodenum. The antioxidant effects of in ovo CGA on postnatal heat-stressed broilers may be related to its regulation of epigenetic mechanisms. Thus, this study provides technical knowledge to support the in ovo feeding of CGA to alleviate oxidative stress in postnatal heat-stressed broilers.

Effect of in ovo creatine monohydrate on hatchability, post-hatch performance, breast muscle yield and fiber size in chicks from young breeder flocks.

Younger broiler breeder flocks produce smaller eggs containing smaller yolks, with potentially lower energy reserves for the developing chick. Creatine is a naturally occurring energy source and is abundant in metabolically active tissues; providing this to chicks in ovo should provide additional energy to improve hatchability and post-hatch growth. Thus, post-hatch performance of male and female chicks hatched from younger breeder flocks supplemented with creatine monohydrate (CrM) in ovo was investigated. Four hundred eggs from Ross 308 breeder hens aged 27 to 29 wk were collected and at d 14 assigned to a treatment group and received 1) no injection, 2) 0.75% saline injection, or 3) 8.16 mg creatine monohydrate in 0.75% saline. At hatch 72 birds (24/treatment) were euthanized and BW, breast muscle, heart and liver weight were obtained, and breast muscle tissue was placed in 10% buffered formalin. Birds were then placed in raised metal pens (24 pens; 10-11 birds/pen; 8 replicates/treatment) and grown to d 42 with BW and pen feed intake measured once a week. At d 42, ninty-six birds were euthanized (2 male and 2 female/pen) and the process occurred as at hatch. Body composition was obtained for 48 birds (2/pen; 1 male,1 female) with a dual energy X-ray absorptiometry (DXA) scanner. Breast muscle tissue was processed for histological analysis and breast muscle fiber parameters were analyzed by ImageJ. While not statistically significant, the CrM treatment group saw an improved hatch rate (CrM: 93.5%, Saline: 88.6%, Control: 88.8%) and reduced early post hatch mortality. Chicks given in ovo CrM had significantly increased creatine concentrations in both liver and heart tissue at hatch compared to those in the saline and control groups. BW, BW gain, and final body composition parameters were not statistically different between treatments and in ovo CrM did not affect breast muscle fiber number or area. The creatine injection likely improved the energy status of the growing embryo resulting in the improved hatch rate but leaving little reserves for post-hatch growth.

The effects of in ovo injection with sulfur amino acids and folic acid on the gene expression, relative organ weights, hematologic parameters, performance, and carcass characteristics of broiler chickens.

This study aimed to evaluate the effect of in ovo injection of folic acid (FA) and sulfur amino acids (SAAs) on the mitotic activity of myocytes, performance, relative organ weight, hematological values, and characteristics of broiler chicken carcasses. A total of 1200 fertile eggs from 42-week-old Ross AP© breeders were inoculated in the albumen on the first day of incubation in a completely randomized design with one of the treatments: C-intact eggs; SS: inoculation with 0.5 mL of saline solution; FA: 0.150 mg of FA; SAA: 5.90 mg of L-methionine and 3.40 mg of L-cysteine; or FA/SAA: FA + SAA. The inoculation of SAA did not influence (p > 0.05) the post-hatching characteristics of the chickens. FA inoculation increased (p < 0.05) the expression of the PAX7 and MYF genes in the pectoralis muscle of hatched chicks and reduced (p < 0.05) feed conversion at 42 days of age. The combination of SAA + FA increased (p < 0.05) the depth of the ileal crypt on the 1st day after hatching and the relative weight of the spleen and thymus on the 21st day of life. In conclusion, the inoculation of FA on the 1st day of incubation increases gene expression and improves the performance of broilers.

Effects of in ovo feeding of vitamin E or vitamin C on egg hatchability, performance, carcass traits and immunity in broiler chickens.

The effect of in ovo feeding of different levels of vitamins C and E on egg hatchability, immune response, growth and carcass traits of broiler chickens were investigated. A total of 672 fertilized eggs were assigned to one of eight experimental groups having three replicates with 28 eggs as follows: (1) negative control (not injected); (2) positive control (injected with 0.2 mL deionized water); (3) vitamin C at 1 mg; (4) vitamin C at 3 mg; (5) vitamin C at 6 mg; (6) vitamin E at 0.5 IU; (7) vitamin E at 0.75 IU; and (8) vitamin E at 1.0 IU. At the end of incubation, the number of chicks hatched, and their individual body weight were recorded. Among hatched birds, a total of 240 mixed chicks were randomly selected (30 subject per group equally shared in three pen floors). Chicks were vaccinated against Avian Influenza, Gumboro, Bronchitis, and Newcastle disease virus. Performance parameters were weekly evaluated until 42 days of age. At days 28 and 42, broiler serum and spleen and Bursa of Fabricius relative weight were assessed as well as on day 42 the carcass traits. From results, in ovo injection with 3 mg of vitamin C or 0.75 IU of vitamin E, increased significantly (p < .05) the embryos hatchability when compared to the negative control. However, body weight at hatch and growth performance parameters showed no differences among treatments. Similarly, in ovo concentrations of vitamins C or E showed no differences on carcass traits, immunity-related organs weight or immune response for anti-Newcastle disease hemagglutination-inhibition and total immunoglobulins against sheep red blood cells (SRBC) when compared to the control groups. Based on findings, it can be concluded that in ovo feeding vitamins E and C supported positively chicken embryos hatchability demonstrating the key-role as antioxidant agents; however, further studies are currently being evaluated.

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.