Sex-specific effects of dietary restriction on physiological variables in Japanese quails.

Nutritional limitation is a common phenomenon in nature that leads to trade-offs among processes competing for limited resources. These trade-offs are mediated by changes in physiological traits such as growth factors and circulating lipids. However, studies addressing the sex-specific effect of nutritional deficiency on these physiological variables are limited in birds. We used dietary restriction to mimic the depletion of resources to various degrees and investigated sex-specific effects on circulating levels of insulin-like growth factor 1 (IGF-1) and triglycerides in Japanese quails (Coturnix japonica) subjected to ad libitum, 20%, 30% or 40% restriction of their daily requirement, for 2 weeks. We also explored the association of both physiological variables with body mass and egg production. While dietary restriction showed no effects on circulating IGF-1, this hormone exhibited a marked sexual difference, with females having 64.7% higher IGF-1 levels than males. Dietary restriction significantly reduced plasma triglyceride levels in both sexes. Females showed more than six-fold higher triglyceride levels than males. Triglyceride levels were positively associated with body mass in females while showed not association in males. Overall, our findings revealed sex-specific expression of physiological variables under dietary restriction conditions, which coincide with body size.

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.

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.

Physiological, antimicrobial, intestine morphological, and immunological effects of fructooligosaccharides in pigs.

In pig nutrition, there are some periods when natural alternatives to antibiotics are more required, such as during suckling and weaning. Fructooligosaccharides (FOSs) are a group of prebiotics applied as feed ingredients in animal nutrition since their positive effects on growth performance, immunological parameters, intestinal microbiota, and gut morphology are reported. Accordingly, FOS may be candidate molecules to improve the mentioned properties in pigs. Previous studies defined FOS as inhibiting the activity of pathogens and increasing the colonization of beneficial bacteria in the gut, although metabolites of FOS decreased the intestinal pH value. Beneficial effects on digestive-enzyme activities and on protein digestion were determined in some studies. All of the three types of FOS (inulin, oligomeric fructans, and short-chain FOSs) promoted the microbial composition of the gut by increasing the colonizations of Lactobacillus, Bifidobacterium, and Prevotella genus. FOS also affected the immune response directly and indirectly and increased vaccine-specific IgA, serum IgG, and IgE levels. Moreover, FOS enhanced the activation of T cells and altered the secretions of some cytokines. Levels of vaccine-specific IgG could not be increased after FOS supplements. In most cases, FOS modified intestinal morphological parameters, such as longer villi, villus-height-to-crypt-depth ratio, and thicker mucosa, which could suggest better absorptive functions. Results are contradictory on growth performance, which might be influenced by the chemical structure, the duration, and the dose of FOS, so further studies are required. This review aims to gather information regarding immunological, antimicrobial, intestine morphological, and growth performance properties of fructooligosaccharides in pigs.

Effect of Carotenoids, Oligosaccharides and Anthocyanins on Growth Performance, Immunological Parameters and Intestinal Morphology in Broiler Chickens Challenged with Escherichia coli Lipopolysaccharide.

This study was conducted to investigate the effect of carotenoid, oligosaccharide and anthocyanin supplementation in broiler diets under Escherichia coli lipopolysaccharide (LPS) challenge. Ross 308 chickens were fed 5 diets: basal diet (control diet), diet supplemented with ß-glucan in 0.05% (positive control) and diets with 0.5% carotenoid-, oligosaccharide- or anthocyanin contents. On the 26th days of age, chickens were challenged intraperitoneally 2 mg LPS per kg of body weight. 12 h after injection, birds were euthanized, then spleen and ileum samples were collected. LPS induced increased relative mRNA expression of splenic (p = 0.0445) and ileal (p = 0.0435) interleukin-1ß (IL-1ß), which was lower in the spleen in carotenoid (p = 0.0114), oligosaccharide (p = 0.0497) and anthocyanin (p = 0.0303)-treated chickens compared to LPS-injected control birds. Dietary supplementation of carotenoids also decreased relative gene expression of splenic interleukin-6 (IL-6) (p = 0.0325). In the ileum, ß-glucan supplementation showed lower relative mRNA expression of toll-like receptor 5 (TLR-5) (p = 0.0387) compared to anthocyanin treatment. Gene expression of both splenic and ileal interferon-α (IFN-α), interferon-γ (IFN-γ), toll-like receptor 4 (TLR-4) and toll-like receptor 5 (TLR-5) were not influenced by dietary supplements. In conclusion, carotenoids, oligosaccharides and anthocyanins could partially mitigate the immune stress caused by LPS challenge. All of the compounds impacted longer villus height (p < 0.0001), villus height:crypt depth ratios were higher after ß-glucan (p < 0.0001) and anthocyanin (p = 0.0063) supplementations and thickened mucosa was observed in ß-glucan (p < 0.0001), oligosaccharide (p < 0.0001) and anthocyanin (p = 0.048) treatments. All of these findings could represent a more effective absorption of nutrients.