The diversification of protein sources used in food formulation has increased the need to assess protein quality beyond traditional food forms. Protein digestibility is a key component of the Protein Digestibility Corrected Amino Acid Score (PDCAAS), the regulatory metric derived from rodent bioassays. Ethical concerns and the high cost of animal testing limit the ability of food formulators to screen protein ingredients and evaluate processing effects on digestibility. Findings from an international collaborative study are presented to position two in vitro methods, the pH-drop and pH-stat assays, as accredited approaches for determining protein digestibility. Nine laboratories participated in the study and analyzed 12 protein ingredients. Relative standard deviations for repeatability ranged from 0.8 to 2.1 % and 0.5-4.8 %, while reproducibility ranged from 1.2 to 3.6 % and 1.1-4.9 % for the pH-drop and pH-stat methods, respectively. Comparison between the two assays demonstrated strong correlation and moderate agreement, with the pH-stat assay yielding slightly lower digestibility values. In vitro protein digestibility coefficients aligned well with literature reported true faecal protein digestibility values for comparable, non-identical protein ingredients. Both methods received official AOCS Uniform Methods Committee approval, offering simple, affordable, reliable, and ethical tools to support informed decisions on protein digestibility during food formulation.

BACKGROUND:Including fish oil (FO) in laying hen diets is an attractive strategy to increase egg and tissue omega (ω)-3 PUFA, while simultaneously protecting hens from potential immune challenges. OBJECTIVES:This study compared effects of a standard hen diet with FO containing EPA and DHA on plasma oxylipins and splenic cytokine expression in LPS-challenged layers. METHODS:Twenty-four 40-wk-old Dekalb White layers were randomly assigned to either control (no EPA or DHA) or FO (0.4 g/100 g EPA and DHA) diets. After 6 wk, hens were challenged with LPS (8 mg/kg body weight; i.v.) or saline 2 h before termination. PUFA were analyzed by GC, free oxylipins by HPLC/MS/MS, and splenic cytokines by quantitative reverse transcriptase-polymerase chain reaction. Data were analyzed by principal component analysis and analysis of variance. RESULTS:Yolk, liver, and plasma were enriched in ω-3 PUFA and reduced in ω-6 PUFA in FO group. Interestingly, LPS increased liver α-linolenic acid (ALA) and plasma ALA and DHA in FO-fed hens (P < 0.05). FO-fed hens exhibited higher plasma EPA and DHA oxylipins, and lower ω-6 oxylipins. Because of an interaction with FO, LPS increased ∼14% of oxylipins in control hens, particularly octadecanoids from ALA and linoleic acid, whereas in FO-fed hens, LPS decreased ∼31% of oxylipins derived from ω-6 and also from ω-3 PUFA (P < 0.05). Consistent with these putatively anti-inflammatory effects, FO decreased oxylipins associated with higher soluble epoxide hydrolase activity in saline-treated hens. However, FO increased these oxylipins in LPS-treated hens (P < 0.05). LPS induced the expression of splenic cytokines, and this was not altered by diet. CONCLUSIONS:Dietary EPA and DHA enrich ω-3 PUFA in yolk, liver, and plasma of layers, and modulate plasma oxylipins. Whether oxylipin changes mediate or are a consequence of FO effects on LPS-induced inflammation remains to be elucidated.

Soybean meals (SBM) SBM ) from different locations differ in their protein content, subsequently influencing their amino acid (AA) AA ) profile. In this study, standardized ileal digestibility (SID) SID ) of AA and growth or production performance were evaluated in pullets and hens fed SBM derived from soybean grown in Western Canada, primarily Manitoba (MB) MB ) labelled as A-, B- and C-SBM compared with that from Eastern Canada (Ontario, ON-SBM) and contained 38.3 f 0.44, 38.6 f 0.61, 39.4 f 0.49, or 44.0 f 0.87% CP, respectively. A Nfree diet was used to determine basal ileal endogenous losses of AA. The study included the grower, developer, and layer phases (9-12, 13-16, and 44/59-64-wk old birds, respectively). Although a lower (P P = 0.029) SID for cysteine was noted in the grower phase for the C-SBM compared with other SBM, the developer phase had higher (P < 0.05) SID for methionine, phenylalanine, cysteine (more by 4.4, 2.4 and 7.2% units, respectively) on average for SBM samples from MB compared with the ON-SBM. Regardless the source of the SBM, no difference in SID of AA was noted in the layer phase. Overall, in all phases the SID values of most AA in the SBM from MB were comparable with the ON-SBM, which may be linked to higher values of these AA per unit of protein content in the former source. In addition, the growth performance including feed intake, BW gain and feed conversion ratio in pullets, and egg production/quality in layers were similar between treatments. These findings show that the MB-SBM have a comparable feeding value with the ON-SBM, hence represent a suitable alternative protein source for poultry.

OBJECTIVE To investigate inflammatory responses to lipopolysaccharide (LPS) injection in layers. ANIMALS 33 40-week-old laying hens were used. METHODS 30 laying hens were divided into 2 groups: the first group was injected with 8 mg/kg LPS, while the second group was injected with sterile saline. At the start of the study, 3 birds served as a baseline and were used as the time 0 controls for both the saline and LPS-treated groups. Blood and spleen tissues were collected at 0 (before) and 1, 2, 3, 4, and 6 hours after injection. RESULTS LPS administration increased splenic mRNA levels of IL-1β, IL-2, IL-6, IL-8, IL-10, interferon-γ, and tumor necrosis factor-α ( P < .001) and serum IL-6 levels ( P < .01) compared to saline injection. The mRNA expression of most cytokine genes increased rapidly toward peak values within 2 hours after the LPS injection, and then the difference between the saline and LPS treatments got smaller as time went on; serum IL-6 reached its highest concentration 2 hours after LPS administration. The magnitude of LPS-induced upregulation of gene expression was the highest for IL-6, followed by IL-1β and IL-8, and tumor necrosis factor-α was the least affected. CLINICAL RELEVANCE The temporal and quantitative profile of these inflammatory mediators generated from this study provides valuable information in identifying the optimal time window and appropriate biomarkers for LPS-induced inflammation, which has significant implications in evaluating the effects of interventions on the immune system of chickens.

BACKGROUND:There is a lack of nutrition guidelines for the feeding of omega-3 polyunsaturated fatty acids (PUFA) to laying hens. Knowledge as to whether the type and concentrations of α-linolenic acid (ALA) and/or docosahexaenoic acid (DHA) in the diet can make a difference to the birds' immune responses when subjected to a lipopolysaccharide (LPS) challenge is limited. OBJECTIVES:The study was designed to determine the potential nutritional and health benefits to laying hens when receiving dietary omega-3 PUFA from either ALA or DHA. METHODS:A total of 80 Lohmann LSL-Classic (white egg layer, 20 wk old) were randomly assigned to 1 of 8 treatment diets (10 hens/treatment), provided 0.2%, 0.4%, 0.6%, or 0.8% of total dietary omega-3 PUFA, provided as either ALA-rich flaxseed oil or DHA-enriched algal biomass. After an 8-wk feeding period, the birds were challenged with Escherichia coli-derived LPS (8 mg/kg; i.v. injection), with terminal sample collection 4 h after challenge. Egg yolk, plasma, liver, and spleen samples were collected for subsequent analyses. RESULTS:Increasing dietary omega-3 supplementation yielded predictable responses in egg yolk, plasma, and liver fatty acid concentrations. Dietary intake of ALA contributed mainly to ALA-derived oxylipins. Meanwhile, eicosapentaenoic acid- and DHA-derived oxylipins were primarily influenced by DHA dietary intake. LPS increased the concentrations of almost all the omega-6 PUFA-, ALA-, and DHA-derived oxylipins in plasma and decreased hepatic mRNA expression of COX-2 and 5-LOX (P < 0.001) involved in the biosynthesis of oxylipins. LPS also increased mRNA expression of proinflammatory cytokine IFN-γ and receptor TLR-4 (P < 0.001) in the spleen. CONCLUSIONS:These results revealed that dietary intake of ALA and DHA had unique impacts on fatty acid deposition and their derived oxylipins and inflammatory responses under the administration of LPS in laying hens.

Context Reducing the environmental impact of animal production is becoming a really hot topic, especially with raised concerns over excessive flows of nitrogen and phosphorus (P) to the environment. Aims The present study was conducted to determine the effects of phytase supplementation on growth, plasma biochemistry, bone mineralisation and P utilisation of pre-lay pullets fed varying levels of non-phytate P. Methods A total of 240 Lohmann pullet chicks were randomly allocated to one of six dietary treatments with eight replicate cages (5 birds per cage) per treatment. Six treatments included three phytase-free diets and three diets supplemented with 1000 U/kg phytase; the non-phytate P levels were 2.75–2.50–2.25, 3.75–3.50–3.25 and 4.75–4.50–4.25 g/kg in the former, and 1.75–1.50–1.25, 2.75–2.50–2.25 and 3.75–3.50–3.25 g/kg in the latter, for the age of 0–4, 4–8 and 8–16 weeks respectively. Key results No significant differences were found for growth performance, plasma biochemistry (calcium, P, alkaline phosphatase and albumin) and bone mineralisation among dietary treatments, but P retention (%) was different (P < 0.001). Analysis of planned contrasts showed that phytase supplementation increased phytate P retention (P < 0.001), and improving the utilisation of phytate P tended most efficiently under low P conditions. Total P retention rate was reduced slightly by phytase supplementation (P < 0.05). Conclusions The results indicated that dietary non-phytate P level could possibly be reduced to 1.75, 1.50 and 1.25 g/kg for 0–4, 4–8 and 8–16 weeks of age respectively after phytase supplementation, without compromising pullet growth and performance during the pre-laying period. Implications The results of this study will contribute to decreasing P excretion by poultry and reducing the potential environmental impact with land application of manure.

Supplementation of n-3 fatty acids to poultry diets is widely acknowledged for its role in enhancing poultry products, however, little is known about the compositional responses of gut microbial communities to type and dosage of these supplements. Here, we compared the effects of n-3 polyunsaturated fatty acids (PUFA), supplied as alpha-linolenic acid (ALA) or docosahexaenoic acid (DHA), on the composition of bacterial communities in ceca of laying hens. Corn-soybean basal diets were supplemented with either flaxseed oil (FO, ALA-rich) or marine algal biomass (MA, DHA-rich), and each supplied 0.20 and 0.60% of total n-3 PUFA in the diet. Lohmann LSL-Classic laying hens (n = 10/treatment) were randomly allocated to one of the 4 diets. After 8 weeks of feeding, blood, liver and cecal digesta samples were obtained for plasma glucose, fatty acids, and short chain fatty acids analyses, respectively. The gut bacterial communities were characterized using genomic DNA ex-tracted from cecal contents, whereby the V3-V4 hypervariable region of the 16S rRNA gene was sequenced using the Illumina Miseq (R) platform. Firmicutes and Bacteroidetes were the predominant phyla in both the FO and MA fed groups. The relative abundance of Tenericutes, often associated with immunomodulation, was relatively higher (P<0.0001) in the FO than MA group. Although the relative abundance of Bacteroides was greater for the FOthan the MA-fed group, this genus was negatively correlated (P<0.05) with total n-3 PUFA in the liver at higher dosages of both FOand MA-fed hens. Higher dose of FO (0.60%) and both dosages of MA (0.20 and 0.60%) substantially enriched several members of Firmicutes (e.g., Faecalibacterium, Clostridium and Ruminococcus) which are known to produce butyrate. Moreover, co-occurrence network analysis revealed that, in the FO 0.60 and MA 0.20-fed hens, Ruminococcaceae was the most influential taxon accounting for about 31% of the network complexity. These findings demonstrate that supplementation of different type and level of n-3 PUFA in hensdiets could enrich microbial communities with potential role in lipid metabolism and health.

Excessive fecal excretion of phosphorus (P) has increasingly become an environmental issue due to oversupply of P in layer rations, and thus it is imperative to minimize safety margins for P to ensure the sustainability of the egg industry. In this study, a 12-week feeding trial (22 to 34 weeks of age) was conducted to evaluate the effects of phytase supplementation on production performance, plasma biochemistry, egg and bone quality and P excretion of laying hens fed various levels of non-phytate P (NPP). Forty-eight Lohmann white laying hens were randomly allocated to one of six corn-soybean meal-oat-based diets: diets containing 2.0, 2.5 or 3.0 g/kg NPP without phytase, and diets containing 1.0, 1.5 or 2.0 g/kg NPP with phytase (1 000 U/kg diet) where phytase inclusion was expected to provide 1.0 g/kg NPP to laying hens, thus making the phytase-unsupplemented treatment served as a control for the phytase-supplemented treatment accordingly. Productive performance was recorded during the experimental period. Blood and egg samples were collected, and digestibility studies were conducted at weeks 6 and 12 of the experiment. Bone mineralization was evaluated at the end of the experiment. Egg weight and egg production, feed consumption, BW and feed conversion ratio of laying hens fed lower NPP diets supplemented with phytase were comparable to those of hens fed high NPP phytase-unsupplemented controls. Eggshell thickness, specific gravity, Haugh units, tibia bone mineral density, tibia ash percent, plasma P and other biochemical parameters were not significantly different among dietary treatments. Total P intake, excretion and retention were affected by diet (P < 0.001), but its deposition in eggs was not significantly different. Contrast analysis further showed that total P excretion of phytase present vs phytase absent was averagely reduced by 40.4 mg/hen per day (P < 0.01). Moreover, total P excretion was linearly (P < 0.01) reduced with lowering dietary NPP, and this relationship was similar regardless of whether phytase was supplemented or not. The results from this study indicated that NPP levels in laying hen diets could be reduced to 1.0 g/kg (excluding the portion of NPP released by phytase) with the inclusion of phytase, without negative effects on production performance and health of the hens, thereby diminishing P excretion into environment. (C) 2020 The Authors. Published by Elsevier Inc. on behalf of The Animal Consortium.

The oversupply of dietary phosphorus (P) leads to increased feed costs and discharge of excessive P to the environment, thus directly impacting the sustainability of egg production practices. The present study was conducted to better define the minimal available P needs of laying hens. Fifty-six Lohmann white laying hens were individually caged and fed one of 7 diets with graded levels of available P (0.15, 0.20, 0.25, 0.30, 0.35, 0.40, or 0.45%) for 12 weeks. Records were maintained for body weight, feed intake, and egg production during the experimental period. Blood and egg samples were collected and digestibility studies conducted at wk 6 and 12 of the experiment. At the end of the experiment, tibia characteristics and expression of the P transporters in the small intestine and kidney were determined. Lowering dietary available P from 0.45 to 0.15% generally reduced plasma P concentrations (P < 0.01), but hen productive performance, plasma calcium, parathyroid hormone and other constituents, tibia bone mineral density and content, tibia ash percentage, and mRNA abundance of the small intestine, and kidney type II sodium/phosphate cotransporter were not different among treatments. Specific gravity and eggshell thickness tended to increase with reducing dietary P (P < 0.05). Total P intake, excretion, and retention net amount decreased (P < 0.01) with reducing dietary P, but its retention rate (of intake) remained unchanged. Furthermore, changing dietary P did not affect calcium retention rate and net deposition of total P and calcium in eggs. These data indicate that reducing dietary available P up to 0.15% is adequate to maintain health and performance of layers. As such, this minimal available P estimate should serve as a benchmark for the assessment of P contents of commercial laying hen rations, with the goal of enhancing the sustainability of egg production.

Dietary phosphorus (P) oversupply is costly to the poultry industry and represents a point source of excessive P discharge to the environment via manure application to soils. The current study was conducted to investigate the effects of dietary available P (AP) level on growth performance, plasma biochemistry, bone characteristics, and P flow in order to assess minimal AP needs of pullets during the pre-lay period. A total of 192 Lohmann pullet chicks were randomly allocated to one of 4 treatments with 8 replicate cages of 6 birds per cage. The pullets of Low (L; the lowest AP regimen) were fed diets in a sequence of 0.200, 0.175, and 0.150% AP for the age period of 0 to 4 wk, 4 to 8 wk, and 8 to 16 wk, respectively. The AP level was increased by an increment of 0.1% in each phase in Low-Medium (LM), Medium-High (MH) and High (H), thus making H (control) consisting of 0.500, 0.475, and 0.450% AP. Overall, there were no main effects of treatment on body weight gain (BWG), feed intake (FI) and feed conversion ratio (FCR). However, compared with the H treatment, BWG or FI of the L treatment was lower during the first 8 wk post-hatch, but became comparable or higher over the last 8 wk (P < 0.05); FCR of L birds was greater over wk 0 to 4 but became smaller over wk 8 to 16 (P < 0.05). Plasma P was lower in L than H treatments at wk 8 but turned comparable at wk 16 (P < 0.001). Bone characteristics were not different for pullets fed various AP regimens. Total P intake and excretion were reduced with lowering dietary AP (P < 0.001), but its retention percent was not affected. The results indicated that the lowest AP regimen (0.200-0.175-0.150%) in the current study was adequate to support healthy growth and development of pre-lay pullets, while reducing P excretion in manure.

This study was conducted to determine the effects of hemp oil (HO) and HempOmega (HΩ), an equivalent product to HO, on performance and tissue fatty acid profile of layers and broiler chickens in two separate experiments. In the first experiment, forty 19-wk old Lohmann white laying hens were randomized to 1 of 5 dietary treatments, either a control diet or a control diet supplemented with 4 or 8% hemp oil provided by HO or HΩ, for a period of 6 wk (n = 8/diet). In experiment 2, 150-day-old mixed-sex (75 male; 75 female) Ross 308 chicks were randomly allocated into 5 dietary treatments, a control diet or a control diet supplemented with either 3 or 6% hemp oil provided by HO or HΩ, each with six replicates of 5 chicks for a 21-d feeding period. Performance of layers and broilers was not affected by dietary treatments. Animals provided with either HO or HΩ diets had greater total n-3 polyunsaturated fatty acids (PUFAs), alpha-linolenic acid (ALA), eicosapentaenoic acid (EPA), docosapentaenoic acid (DPA), and docosahexaenoic acid (DHA) in egg yolks, thighs, and breasts compared to the control diet (P < 0.01), and monounsaturated fatty acids (MUFAs) content of egg yolks and thighs decreased (P < 0.05). The levels of total n-6 PUFAs, linoleic acid (LA), or arachidonic acid (ARA) of the egg yolk and meat were generally not affected by dietary supplementation with HO or HΩ, but gamma-linolenic acid (GLA) was notably increased (P < 0.01). The current data show that inclusion of hemp oil up to 8% in layer diets and 6% in broiler diets provided by HO or HΩ does not negatively affect overall performance of birds and results in the enrichment of n-3 PUFAs and GLA in eggs and meat.

The present study was conducted to investigate the effects of dietary folic acid (FA) supplementation on performance, serum biochemical indices, and mRNA abundance of intestinal folate transporters in young and older laying hens after acute lipopolysaccharide (LPS) challenge. Two experiments were conducted separately involving 48 Shaver White young laying hens (24 wk of age) in experiment 1 and 48 Shaver White older laying hens (58 wk of age) in experiment 2. Birds were fed 2 diets in a complete randomized design. The diets were wheat-soybean meal based, with or without supplemental 4 mg of FA/kg of diet. Birds were fed for 8 wk, during which time feed consumption and egg production were monitored. At the end of each feeding experiment, 6 hens from each dietary treatment were injected intravenously with 8 mg/kg of BW of either Escherichia coli LPS or sterile saline. Four hours after injection, blood and intestinal samples were collected for further analysis. Compared with the control, dietary FA supplementation increased egg weight and egg mass and decreased serum glucose levels in the young laying hens, and reduced serum uric acid in the older laying hens (P < 0.05). Relative to saline injection, plasma homocysteine, serum calcium, and phosphorus levels were found to be lower in both young and older laying hens after LPS challenge (P < 0.05). Other serum biochemical variables and the mRNA expression of 2 folate transport genes in the small and large intestine were differentially affected by LPS challenge, and some of those responses varied with the age of the birds. Additionally, interactions between diet and LPS challenge were specifically found in the older laying hens. In summary, in addition to improving production performance, there were effects of dietary FA supplementation and its interaction with LPS challenge on biochemical constituents, and age played a role in the development of responses to diet and bacterial LPS infections.

Methionine synthase (MS) and betaine-homocysteine methyltransferase (BHMT) are both zinc (Zn)-dependent methyltransferases and involved in the methylation of homocysteine. The objective of this study was to investigate the effects of dietary Zn supply on homocysteine levels and expression of the two enzymes in growing rats. Male weanling Sprague-Dawley rats were assigned randomly to four dietary groups (n=8/group) for 3 weeks: Zn deficient (ZD; <1mg Zn/kg); Zn control (ZC; 30mg Zn/kg); Zn supplemented (ZS; 300mg Zn/kg); pair fed (PF; 30mg Zn/kg) to the ZD group. Serum and femur Zn concentrations were 83% and 58% lower in ZD, and 49% and 62% higher in ZS compared to ZC (P<0.001), respectively. The ZD rats had lower feed intake (37%), body weight gains (45%), liver (43%) and kidney (31%) weights than those of ZC (P<0.001), but these parameters in ZD were not significantly different from the PF controls. Serum homocysteine concentrations were 65% higher in ZD compared to PF (P<0.05), and there was no significant difference in serum folate levels between ZD and PF groups. The mRNA expression of liver and kidney MS was 57% and 38% lower in ZD than PF (P<0.001), respectively. Hepatic and renal BHMT mRNA levels were not altered in ZD compared to controls. The aforementioned measurements were not significantly different between ZS and ZC groups, except Zn levels. These results demonstrated that homocysteine homeostasis appeared to be disturbed by Zn deficiency but not Zn supplementation, and elevated serum homocysteine might be due to reduced expression of MS during Zn deficiency.

Folic acid plays a key role in nucleic acids and protein synthesis, and has been associated with anti-inflammatory effects in LPS-induced infections. To this end, a study was conducted to investigate the effects of dietary folic acid (FA) supplementation in older laying hens (58 to 66 wk of age) challenged with Escherichia coli lipopolysaccharide (LPS). A total of 24 Shaver White laying hens at 58 wk were fed 2 diets. The diets were wheat-soybean-based, with either 0 or 4 mg of supplemental FA per kg of diet. After 8 wk of feeding and at 66 wk, the hens were injected intravenously with 8 mg of LPS or saline per kg of BW. Four hours after injection, blood was collected and hens were euthanized to obtain spleen and cecal tonsils. The T cell subsets in the blood and the spleen (CD4+ and CD8+), total IgG, and biochemical constituents (total protein, albumin, globulin, and fibrinogen) were not influenced (P > 0.05) by dietary FA supplementation. However, LPS injection decreased (P < 0.05) biochemical constituents, CD4+, and CD8+ cells in the blood, whereas CD4+:CD8+ ratio and total IgG increased (P < 0.05), and fibrinogen was not influenced. Gene expression in the spleen and cecal tonsils was not influenced by dietary FA supplementation except a diet × challenge interaction for interleukin (IL)-8 in the spleen; IL-8 decreased in FA-fed hens that were treated with LPS. Also, FA supplementation decreased the expression of IL-8 in cecal tonsils. Relative to saline-injected hens, expression of IL-1β, interferon-γ, and IL-10 increased in the LPS-injected hens in the spleen and cecal tonsils, IL-8 increased in LPS-injected hens only in the cecal tonsils, whereas Toll-like receptor 4, IL-4, IL-17, and IL-18 increased in the LPS-injected hens only in the spleen; however, LPS decreased expression of IL-13 in the cecal tonsils. In conclusion, FA did not affect inflammatory responses in older laying hens; more studies are required to investigate possible protective effects of FA in laying hens.

Effects of diet and ontogeny on the expression of fatty acid desaturases and elongases were examined in broiler chickens. In Study 1, 120 day-old male chicks received one of six diets with LA:ALA ranging from 46:4 to 16:34, for 33 days. Total n-6 PUFA decreased, and n-3 PUFA increased in response to a decrease in the dietary LA:ALA. FADS1, FADS2, ELOVL2 and ELOVL5 mRNAs were highest (P<0.05) in birds fed lower LA:ALA diets. In Study 2, 60 day-old male chicks were fed a basal diet, and liver samples were collected on day of hatch, and on days 2, 7, 14, 21 and 35 post-hatch. Total n-6 and n-3 PUFA increased (P<0.01) from days 7 to days 21. FADS1, FADS2 and ELOVL2 mRNAs generally increased (P<0.01) with age. These findings provide evidence for the dietary and developmental regulation of PUFA metabolism in broiler chickens.

Folate transporters, including the reduced folate carrier and the proton-coupled folate transporter, encoded by Slc19a1 and Slc46a1 genes respectively, play important roles in the transport of folate across biological membranes given the hydrophilic nature of folates. Although a number of studies have demonstrated that these two transporters are regulated ontogenetically in mammals, little data are available on their developmental patterns of expression in poultry. The objective of this study was to investigate the expression patterns of Slc19a1 and Slc46a1 in jejunal and cecal tissue of broiler chickens during post-hatching development. Post-hatch male chicks (Ross × Ross) had free access to water and a soybean/wheat-based diet. Jejunal, cecal and blood samples were collected on day-of-hatch but before feeding (D0), and on D2, D7, D14, D21 and D35 post-hatch (n = 8 at each time point), respectively. Plasma folate concentrations were low on the day of hatch and increased with maturation; by contrast, plasma homocysteine, a marker of folate status, was highest (P < 0.05) in the day-of-hatch birds and decreased thereafter. Increasing age reduced mRNA abundance of Slc19a1 (P < 0.05) in the jejunum and cecum. Abundance of Slc46a1 mRNA (P < 0.05) gradually decreased in the cecum with increasing age and that of Slc46a1 in the jejunum initially decreased and then increased to level similar to that of day-of-hatch. The study provides some initial data on ontogenetic regulation of Slc19a1 and Slc46a1 in the jejunum and cecum of the chicken and lays the ground work for future nutritional studies. Moreover, the expression of Slc19a1 and Slc46a1 transcripts in the cecum provides evidence of the potential for cecally derived folate to contribute to the folate status of the host.

We investigated the effects of dietary folic acid (FA) supplementation on immunological parameters in young laying hens under acute conditions of lipopolysaccharide (LPS) challenge. Twenty-four Shaver White laying hens at 24 wk were fed 2 diets in a completely randomized design. The diets were wheat-soybean based, with either 0 or 4 mg of supplemental FA per kilogram of diet. At 32 wk of age, 6 hens from each dietary treatment were injected intravenously with 8 mg/kg of BW of either LPS or saline. Four hours after injection, blood was collected and the hens were euthanized to obtain spleen and cecal tonsils. Heterophil:lymphocyte ratio, CD3+, CD4+, CD8+ T cells, and CD4+:CD8+ cells in the blood and spleen were not affected by dietary FA. Relative to saline-injected hens, LPS-injected hens had fewer (P < 0.05) CD3+, CD4+, CD8+, and CD4+:CD8+ cells in the blood, and no difference was found in the spleen. Total protein, albumin, and globulin were found to be higher (P < 0.05) in FA-supplemented hens compared with the control. However, total protein, albumin, and globulins decreased (P < 0.05) in the LPS-injected hens compared with the saline control. Expression of interleukin (IL)-1β in cecal tonsils decreased (P < 0.05) in FA-supplemented hens, but no dietary influence was found on the expression of other genes in both the spleen and cecal tonsils. Lipopolysaccharide upregulated (P < 0.05) expression of IL-10 and interferon (IFN)-γ in the spleen, and IL-1β, IL-10, and IFN-γ in the cecal tonsils, whereas the expression of Toll-like receptor (TLR)-4 and IL-8 was not influenced by LPS in the spleen and cecal tonsils. There was a diet × challenge interaction for total IgG, and cytokines IL-1β and IL-18 in the spleen as well as IL-18 in the cecal tonsils. In conclusion, there were few interactions of dietary FA and LPS; however, FA increased biochemical constituents, enhanced generation of total IgG, as well as exhibiting pleoitropic effects in inflammatory responses.