Dietary restriction and life-history trade-offs: insights into mTOR pathway regulation and reproductive investment in Japanese quail.

Resources are needed for growth, reproduction and survival, and organisms must trade off limited resources among competing processes. Nutritional availability in organisms is sensed and monitored by nutrient-sensing pathways that can trigger physiological changes or alter gene expression. Previous studies have proposed that one such signalling pathway, the mechanistic target of rapamycin (mTOR), underpins a form of adaptive plasticity when individuals encounter constraints in their energy budget. Despite the fundamental importance of this process in evolutionary biology, how nutritional limitation is regulated through the expression of genes governing this pathway and its consequential effects on fitness remain understudied, particularly in birds. We used dietary restriction to simulate resource depletion and examined its effects on body mass, reproduction and gene expression in Japanese quails (Coturnix japonica). Quails were subjected to feeding at 20%, 30% and 40% restriction levels or ad libitum for 2 weeks. All restricted groups exhibited reduced body mass, whereas reductions in the number and mass of eggs were observed only under more severe restrictions. Additionally, dietary restriction led to decreased expression of mTOR and insulin-like growth factor 1 (IGF1), whereas the ribosomal protein S6 kinase 1 (RPS6K1) and autophagy-related genes (ATG9A and ATG5) were upregulated. The pattern in which mTOR responded to restriction was similar to that for body mass. Regardless of the treatment, proportionally higher reproductive investment was associated with individual variation in mTOR expression. These findings reveal the connection between dietary intake and the expression of mTOR and related genes in this pathway.

Dietary restriction reveals sex-specific expression of the mTOR pathway genes in Japanese quails.

Limited resources affect an organism's physiology through the conserved metabolic pathway, the mechanistic target of rapamycin (mTOR). Males and females often react differently to nutritional limitation, but whether it leads to differential mTOR pathway expression remains unknown. Recently, we found that dietary restriction (DR) induced significant changes in the expression of mTOR pathway genes in female Japanese quails (Coturnix japonica). We simultaneously exposed 32 male and female Japanese quails to either 20%, 30%, 40% restriction or ad libitum feeding for 14 days and determined the expression of six key genes of the mTOR pathway in the liver to investigate sex differences in the expression patterns. We found that DR significantly reduced body mass, albeit the effect was milder in males compared to females. We observed sex-specific liver gene expression. DR downregulated mTOR expression more in females than in males. Under moderate DR, ATG9A and RPS6K1 expressions were increased more in males than in females. Like females, body mass in males was correlated positively with mTOR and IGF1, but negatively with ATG9A and RS6K1 expressions. Our findings highlight that sexes may cope with nutritional deficits differently and emphasise the importance of considering sexual differences in studies of dietary restriction.

Embryonic methionine triggers post-natal developmental programming in Japanese quail.

Embryonic development is one of the most sensitive and critical stages when maternal effects may influence the offspring's phenotype. In birds and other oviparous species, embryonic development is confined to the eggs, therefore females must deposit resources into the eggs to prepare the offspring for the prevailing post-natal conditions. However, the mechanisms of such phenotypic adjustments remain poorly understood. We simulated a maternal nutritional transfer by injecting 1 mg of L-methionine solution into Japanese quail eggs before the onset of incubation. The increase in early methionine concentration in eggs activated the insulin/insulin-like signalling and mechanistic target of rapamycin (IIS/mTOR) signalling pathways and affected post-natal developmental trajectories. Chicks from methionine-supplemented eggs had higher expression of liver IGF1 and mTOR genes at hatching but were similar in size, and the phenotypic effects of increased growth became apparent only a week later and remained up to three weeks. Circulating levels of insulin-like growth factor-1 (IGF-1) and expression of ribosomal protein serine 6 kinase 1 (RPS6K1), the mTOR downstream effector, were elevated only three weeks after hatching. These results show that specific nutritional cues may have phenotypic programming effects by sequentially activating specific nutrient-sensing pathways and achieving transgenerational phenotypic plasticity.

Methionine sources and genotype affect embryonic intestinal development, antioxidants, tight junctions, and growth-related gene expression in chickens.

Methionine (Met) is an essential and first limiting amino acid in the poultry diet that plays a significant role in chicken embryonic development and growth. The present study examined the effect of in ovo injection of DL-Met and L-Met sources and genotypes on chicken embryonic-intestinal development and health. Fertilized eggs of the two genotypes, TETRA-SL layer hybrid (TSL) - commercial layer hybrid and Hungarian Partridge colored hen breed (HPC) - a native genotype, were randomly distributed into four treatments for each genotype. The treatment groups include the following: 1) control non-injected eggs (NoIn); 2) saline-injected (SaIn); 3) DL-Met injected (DLM); and 4) L-Met injected (LM). The in ovo injection was carried out on 17.5 d of embryonic development; after hatching, eight chicks per group were sacrificed, and the jejunum was extracted for analysis. The results showed that both DLM and LM groups had enhanced intestinal development as evidenced by increased villus width, villus height, and villus area (P < 0.05) compared to the control. The DLM group had significantly reduced crypt depth, glutathione content (GSH), glutathione S-transferase 3 alpha (GST3), occludin (OCLN) gene expression and increased villus height to crypt depth ratio in the TSL genotype than the LM group (P < 0.05). The HPC genotype has overexpressed insulin-like growth factor 1 (IGF1) gene, tricellulin (MD2), occludin (OCLN), superoxide dismutase 1 (SOD1), and GST3 genes than the TSL genotype (P < 0.05). In conclusion, these findings showed that in ovo injection of Met enhanced intestinal development, and function, with genotypes responding differently under normal conditions. Genotypes also influenced the expression of intestinal antioxidants, tight junction, and growth-related genes.

In ovo feeding of methionine affects antioxidant status and growth-related gene expression of TETRA SL and Hungarian indigenous chicks.

Methionine (Met) plays a substantial role in poultry due to its involvement in several pathways, including enhancing antioxidant status and improving growth performance and health status. This study examined how in ovo feeding of Met affects hatching performance, antioxidant status, and hepatic gene expression related to growth and immunity in the TETRA-SL LL hybrid (TSL) commercial layer and Hungarian partridge colored hen (HPC) indigenous genotypes. The eggs were injected with saline, DL-Met, and L-Met on 17.5 days of embryonic development. The results showed that the in ovo feeding of DL-Met significantly increased the hatching weight and ferric reducing the ability of the plasma (FRAP) compared with L-Met. The in ovo feeding of either Met source enhanced the liver health and function and hepatic antioxidant status of the chicks. The genotype's differences were significant; the TSL genotype had better hatching weight, an antioxidant defense system, and downregulated growth-related gene expression than the HPC genotype. In ovo feeding of either Met source enhanced the chicks' health status and antioxidant status, and DL-Met improved the hatching weight of the chicks more than L-Met. Genotype differences were significantly evident in the responses of growth performance, antioxidant status, blood biochemical parameters, and gene expression to Met sources.

Assessment of the microbiological status of two Hungarian ostrich farms.

The aim of this study was to estimate the occurrence of bacterial infection and contamination in two ostrich-producing farms. Compared to other poultry species, the hatchability of ostrich eggs is especially low. In a quest to identify factors that may affect hatchability, we collected faecal samples from adult birds, as well as eggs with dead-in-shell embryos, dead chicks and swab samples from the surface of the eggs and from the environment. The samples were screened for the presence of bacteria by routine bacteriological culture methods. The most prevalent bacteria, detected in the samples, were Escherichia coli, Bacillus spp. and coliform bacteria, whereas Pseudomonas spp. were less frequently found. The intensity and species compositon of the bacterial contamination was comparable in the two farms. Our results revealed that the bacteria, present in the environment, may likely be transmitted to the surface of the eggs. If they are able to penetrate the shell then the embryos and chicks become infected easily. These findings draw the attention to the special importance of enforcing efficient decontamination and disinfection measures to keep the environment and egg surface free from germs. Besides the appropriate egg treatment procedure, the incubation and hatching technology should also be kept under control.

Capsanthin supplementation modulates the immune response in broiler chickens under Escherichia coli lipopolysaccharide challenge.

Due to the legislation of antibiotic usage, natural substances are required for application in the poultry industry. Because of their potential anti-inflammatory and immunomodulatory effects, carotenoids are great sources. Capsanthin, a major carotenoid giving the red color of pepper, is a promising feed additive, as it can reduce chronic inflammation. This study was conducted to determine the effects of capsanthin supplementation at 80 mg kg-1 in feed on the immune response of broiler chickens under Escherichia coli O55:B5 lipopolysaccharide (LPS) challenge. Ross 308 male broilers were divided into treatments: control (basal diet) and feed-supplemented groups. At 42 d of age, chickens were weighed and then challenged with 1 mg LPS per kilogram of body weight intraperitoneally. Four hours after injection, birds were euthanized, and then spleen and blood samples were collected. Capsanthin supplement at 80 mg kg-1 did not change the growth parameters and the relative spleen weight. LPS immunization resulted in higher splenic interleukin-1ß (IL-1ß), interleukin-6 (IL-6), and interferon-γ (IFN-γ) mRNA expressions. Capsanthin addition reached lower gene expression levels of IL-6 and IFN-γ compared to the LPS-injected birds. At plasma level, dietary capsanthin resulted in lower IL-1ß and IL-6 levels. These results may indicate the potential anti-inflammatory effect of capsanthin supplementation in broiler chickens.

Effects of DL and L-Methionine on Growth Rate, Feather Growth, and Hematological Parameters of Tetra-SL Layers from 1-28 Days of Age.

The study was carried out to determine whether sources or levels of methionine (Met) affect the health status of Tetra-SL (TSL) chicks by examining growth performance, feather growth, and hematological parameters. A total of ninety-six (96) day-old (44.2 ± 0.18 g lw) TSL chicks were randomly allotted to six treatment groups (three levels for each DL and L-Met source) with four replicates of four chicks each. Chicks were fed ad libitum diets supplemented with 90, 100, and 110% of methionine requirements for four weeks after hatch. The parameters examined are bodyweight (BW), average daily gain (ADG), feather length (FL), and hematological parameters, including: red blood cell (RBC) count, hemoglobin (Hb) concentration in the blood, hematocrit (Ht; %), number of white blood cells (WBC), platelet count, mean corpuscular volume of red blood cells (MCV), mean corpuscular hemoglobin (MCH), mean corpuscular hemoglobin concentration (MCHC), lymphocyte (LYM), mid-range (MID), and granulocyte (GRAN). There was no significant effect of Met sources and levels on BW, ADG, and FL of TSL chicks for the first four weeks of rearing. The RBC, Hb, Ht, WBC, LYM, MID, and GRAN values of TSL chicks were statistically influenced by dietary Met sources and Met levels (p < 0.05). Among the treatment groups, the number of white blood cells (WBC) on 90% DL−Met was the lowest. WBC, RBC, Hb, and Ht were higher from chicks that received 100% DL-Met than all other levels, regardless of the Met sources. The MCV, MCH, and MCHC values were not affected by either Met source or levels or their interactions. Met source and level interactively affected the Ht, WBC, LYM %, and GRAN values of TSL chicks (p < 0.05). The platelet number was affected by Met source only (p < 0.05) with chicks receiving L-Met source having more than twofold higher platelet values than DL-Met source. In conclusion, varying Met levels by ±10% of the requirement does not adversely affect the growth performance, feather growth, and hematological parameters of TSL chicks reared for up to four weeks of age. DL-Met increased the body weight and improved the white blood cells, red blood cells, and hematocrit at 28 days after hatch. DL-Met showed similar biological efficacy as L-Met for body weight and feather growth but not for the hematological parameters.