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This study evaluated the effects of feeding varying levels of dietary DL-methionine (MET) supplementation on breast meat (BM) quality of broilers of different sex. The 1-day-old, sexed chicks (Ross 708, 1552) were randomly allocated into four groups (each with 4 replicates) and were raised with diets supplemented with 0, 0.5, 1, or 2 g MET/kg of feed to a common weight (2.72 kg). Color, pH, drip loss (DL), water-holding capacity, moisture uptake, cooking yield (CY), texture, total antioxidant capacity (TAC), and lipid oxidation (LO) were determined using BM samples harvested 24 h postmortem. The male BM had higher redness, TAC, firmness, and toughness but lower yellowness (p < 0.01) than the female BM. In both sexes, birds fed 0.5 g/kg MET had lower DL (p < 0.01) than those fed 1 and 2 g/kg MET. For storage up to 3 days, MET suppressed LO in cooked BM (p < 0.01) and suppression increased as MET increased. CY for 1 and 2 g/kg MET were higher (p < 0.01; 79.04 and 78.60%, respectively) than CY for 0 and 0.5 g/kg MET (66.18 and 68.03%, respectively). These results suggest that MET supplementation at 1 g/kg or higher for broilers can improve oxidative stability, muscle functionality, and breast meat CY.

A study was conducted to evaluate the effect of dietary 25-hydroxyvitamin D3 (25OHD) on pullet and egg-laying hen growth performance, egg production, and egg quality. Three hundred and ninety 1-day-old Hy-Line W36 pullets were randomly allocated to 3 treatments with 10 replicated cages and 13 birds per cage. Dietary treatments were vitamin D3 at 2,760 IU/kg (D); vitamin D3 at 5,520 IU/kg (DD), and vitamin D3 at 2,760 IU/kg plus 25OHD at 2,760 IU (69 μg)/kg (25D). Body weight and feed intake were recorded at the end of each stage: starter 1 (0–3 wk), starter 2 (4–6 wk), grower (7–12 wk), developer (13–15 wk), prelay (15–17 wk), peaking (18–38 wk), layer 2 (39–48 wk), layer 3 (49–60 wk), layer 4 (61–75 wk), and layer 5 (76–95 wk). Egg production was recorded daily. Egg quality was evaluated every 8 wk starting from 25 wk. There was no difference in growth performance during the rearing period (0–17 wk). In the laying period (18–95 wk), DD showed lower feed intake at layer 2, but higher intake at layer 3 along with lower hen day production (HDP) from 22 to 48 wk compared to the other treatments. During the same period, the DD group laid smaller eggs with higher specific gravity and shell thickness compared with the other treatments or D alone at 40 wk, which may be partly due to the lower body weight. In contrast, 25D had better feed conversion ratio (feed intake per dozen of eggs) at layer 2, and higher overall (22–60 wk) HDP compared with DD. For the egg quality analysis, at 25 and 33 wk, both DD and 25D had higher Haugh unit compared with D. However, 25OHD has no effects on eggshell quality during the entire production period and no beneficial effects on egg production during the later laying period (after 60 wk). In summary, long-term and early supplementation of 25OHD has positive effects on egg production and egg quality, and the beneficial effects were mainly observed during the early laying stage.

The application of micro-CT in small animal research, especially on bone health, has risen exponentially in recent years. However, its application in egg-laying hen bone analysis was still limited. This review introduces the technical aspects of micro-CT in egg-laying hen bone analysis, especially with the medullary bones presented in the cavity. In order to acquate application of micro-CT for laying hen bone research, image acquisition, reconstruction, and analysis settings need to be adjusted properly. The key difference regarding the application of micro-CT in laying hen bone compared to other small animals such as mice and rats was the larger bone size and more complex structures of medullary and trabecular bones. In order to analyze the details of laying hen bone structures, the volume of interest for laying hen should be selected at a region where all 3 bones are present (critical, trabecular, and medullary bone). Owing to the complexity of bone structures, the conventional techniques are not useful to distinguish the trabecular bone and medullary bone in laying hens accurately. In the current review, an automated segmentation algorithm is described to allow researchers to segment bone compartments without human bias. The algorithm is designed according to the morphology difference of medullary bones compared to trabecular and cortical bones. In this procedure, the loosely woven bones were separated by applying dual thresholds. The medullary calcium chunks were separated by opening or closing procedures, where we defined the diameter of medullary chunks being higher than the trabecular bone thickness as a separation trait. The application of micro-CT in laying hen bone health assessment will significantly expand our understanding of chicken bone physiology and osteoporosis, contributing to improve welfare in laying hens.

Egg-laying hens have a unique bone development pattern due to the medullary bone formation and high bone turnover rate. The role of long-term supplementation of an intermediate form of vitamin D3, 25-hydroxyvitamin D3 (25OHD), on skeletal development of pullets and laying hens is not well established. Exploring its effects on layer bone development will help develop a strategy for preventing laying hen osteoporosis. The purpose of this study was to investigate the role of long-term supplementation of 25OHD in layer diets on bone 3-dimensional structural development. A total of 390 1-day-old Hy-Line W36 pullets were randomly allocated to 3 treatments with 10 replicate cages and 13 birds/cage. Dietary treatments were 1) vitamin D3 at 2,760 IU/kg, 2) vitamin D3 at 5,520 IU/kg, and 3) vitamin D3 at 2,760 IU/kg plus 25OHD at 2,760 IU (69 μg)/kg. The level of 25OHD in the serum was tested throughout the whole experimental period (0–95 wk). Bone growth rate (BGR) was measured at 10 wk using a calcein injection technique. Femurs were scanned using Micro-CT for 3D structural analysis, and the whole-body composition analysis was performed using a dual-energy x-ray absorptiometry method at 17, 60, and 95 wk. Dietary supplementation of 25OHD significantly increased 25OHD level in the serum from 0 to 95 wk. During the rearing period (0–17 wk), 25OHD increased BGR, cortical tissue volume, and bone marrow area at 17 wk, simultaneously. 25OHD created more pores in cortical bone, which resulted in a lower cortical bone mineral density (BMD) but without alerting bone mineral content (BMC). This effect allowed more space for mineral deposition in bones during the later egg-laying period. At 60 wk, the 25OHD group had significantly greater BMD, which led to the highest total BMC, cortical volume, and trabecular bone connectivity. At 95 wk, the birds fed 25OHD had significantly higher cortical bone volume and lower porosity. The 25OHD group also had higher total BMD and medullary bone volume but a lower BMC and volume of trabecular bone than vitamin D3 or double dosage vitamin D3 treatment. This indicated that the bone resorption rate was lower in cortical bone than that in trabecular bone in the late laying period. In conclusion, supplementation with dietary 25OHD could stimulate bone growth and increase bone volume in pullets to provide more space for mineral deposition during the laying period with positive effects on laying hen bone quality.

The objective of present study was to investigate the effects of alpha-lipoic acid (ALA) dietary supplementation on the behavior, physiological and oxidant stress indicators, and bone quality in broilers under high stocking density (HSD) stress. A total of one thousand eight hundred 22-day-old Arbor Acres male broiler chicks were randomly allocated to 18 pens (2.97 × 2.03 m) in 3 groups: 14 birds/m2 (NSD, normal stocking density) or 18 birds/m2 (HSD) or 18 birds/m2 plus 300 mg/kg ALA dietary supplement (HSD + ALA, high stocking density + alpha-lipoic acid). Each treatment had 6 replicates, and the experiment lasted 3 wk. The HSD group was significantly lower than the NSD group (P 0.05) in the standing and preening behavior, alkaline phosphatase activity, PTH concentration, H/L ratio, CAT, T-AOC, MDA, SOD, and GSH-Px. These results indicate that the increase of stocking density lowered the bone quality, increased the physiological and oxidative stress indicators, and modified the behavior of broilers, whereas ALA dietary supplementation could counteract the reduction in the performance and physiological responses of broilers under high-density environmental stress.

Broiler breeders are commonly feed restricted using some variation of skip-a-day feeding to prevent excessive body weight (BW) gain and poor flock uniformity that results in lower production levels. However, the level of feed restriction has increased leading to negative effects on broiler breeder welfare. Research needs to be conducted to evaluate alternative feeding programs to diminish the negative impact of restricted feeding on bird welfare. This research examined pullets that were fed soybean hulls (alternate day feeding, ATD) on the off day of a traditional skip-a-day feeding program in comparison to the standard skip-a-day program (SAD). The 2 dietary feeding treatments each had 3 replicate pens of 210 pullets each and were fed from wk 5 to 21 of age. Nitrogen-corrected true metabolizable energy and digestible amino acid coefficients of soybean hulls were determined. Body weight in the ATD feed program was significantly higher (P < 0.001) than the birds on the SAD feed program until 16 wk of age. Feed allocations for the SAD feeding program was increased at 11 wk of age to achieve similar BW prior to photo stimulation. The ATD feed program significantly improved BW uniformity of the birds for weeks 8, 12, 16, and 20. Hens fed on the SAD feed program had a lower mean egg production than the hens fed on the ATD program. There were significant differences on plasma corticosterone concentrations between the feeding days (24 or 48 h after feeding) in both feed programs. There was a shift in the behavior of the birds with significant differences in the feeding, foraging, and comfort behaviors between the feeding programs on the same feed day. Overall, feeding the ATD females soybean hulls on the off feed day improved the BW uniformity and egg production, but further research will be needed to determine potential differences in nutrient utilization or behavior of the pullets that positively impacted this flock performance.

This study was designed to evaluate the effects of dietary supplemental DL-methionine (MET) on live performance and meat yield for broilers raised to a common weight. A total of 1552 one-day old Ross 708, sexed broilers were randomly distributed to 32 pens resulting in eight treatments (TRT) of four replicates with 44 male or 53 female/pen. A randomized complete block with a 2 × 4 (sex × 4 MET levels 0, 0.5, 1, and 2 g/kg) factorial arrangement of TRT was used. A common weight of 2400 g was approached by day 46 (1 and 2 g MET/kg feed) and day 48 (0 and 0.5 g MET/kg feed). Supplementation of MET at 1, and 2 g/kg had a lower (p &lt, 0.01) feed conversion ratio (FCR) at day 46/48 than broilers fed 0.5 g MET/kg. Broilers without supplemental MET had the worst (p &lt, 0.01) feed conversion and average daily gain (ADG) at day 46/48. Birds fed 0 g MET/kg of feed had lower (p &lt, 0.05) whole eviscerated carcass without giblets (WOG), yield than birds fed 2 g MET/kg of feed. Additionally, birds fed 0 g MET/kg of feed had lower (p &lt, 0.05) breast fillet and tender percent yields than birds fed supplemental MET. Elimination of MET from organic broiler diets resulted in reduced ADG, breast fillet yield and feed efficiency of meat yield of broilers raised to day 46/48. Reduction in MET supplementation below current levels reduced the efficiency of meat production of organic broilers raised to day 46/48.

Changes in medullary and cortical bone structure with age remain unclear. Twenty Hy-Line W36 hens, 25 or 52 weeks of age, were euthanized, and both tibiae were collected when an egg was present in the magnum. Serial cross sections of the tibiae were stained with Alcian blue. The bones were scanned using micro-computed tomography. Trabecular width (Tb.Wi) was significantly higher (p &lt, 0.05) in 25-week-old hens, whereas medullary bone tissue volume (TV) was significantly higher (p &lt, 0.01) in 52-week-old hens. 25-week-old hens had significantly higher (p &lt, 0.01) bone volume fraction (BVF = calcified tissue / TV). Moreover, the cortical bone parameters were significantly higher (TV and bone mineral content (BMC) at p &lt, 0.05, and bone volume (BV) and BVF at p &lt, 0.01) in younger hens. Open porosity and total porosity, which indicate less density, were significantly higher (p &lt, 0.01) in older hens. Older hens showed significantly higher (p &lt, 0.01) tibial diaphysis TV than younger hens. Younger hens had significantly higher (p &lt, 0.01) BV, BVF and bone mineral density (BMD) of the tibial diaphysis. These findings reveal that reductions in medullary bone quality might be associated with age-related low estrogen levels and stimulation of osteoclastic bone resorption by parathyroid hormone. Cortical bone quality decreased with enlargement of the Haversian canals and loss of volume, with a longer egg-laying period leading to osteoporosis.

1,25-dihydroxyvitamin D3 (1,25OHD) has been suggested to play an important role in osteogenic differentiation and mineralization. However, limited data have been reported in avian species. In the present study, the direct role of 1,25OHD on osteogenic differentiation and mineralization in chicken mesenchymal stem cells (cMSCs) derived from day-old broiler bones was investigated. cMSCs were treated with control media (C), osteogenesis media (OM), OM with 1, 5, 10, and 50 nM 1,25OHD, respectively. The messenger RNA (mRNA) samples were obtained at 24 and 48 h and 3 and 7 days to examine mRNA expression of key osteogenic genes [runt related transcription factor 2 (RUNX2), bone morphogenetic protein 2 (BMP2), collagen type I alpha 2 chain (COL1A2), bone gamma-carboxyglutamate protein (BGLAP), secreted phosphoprotein 1 (SPP1), and alkaline phosphatase (ALP)]. Cells were stained at 7, 14, and 21 days using Von Kossa (mineralization), Alizarin Red (AR; mineralization), and Alkaline Phosphatase (early marker) staining methods. From the mRNA expression results, we found a time-dependent manner of 1,25OHD on osteoblast differentiation and mineralization. In general, it showed an inhibitory effect on differentiation and mineralization during the early stage (24 and 48 h), and a stimulatory effect during the late cell stage (3 and 7 days). The staining showed 1,25OHD had an inhibitory effect on ALP enzyme activities and mineralization in a dosage-dependent manner up to 14 days. However, at 21 days, there was no difference between the treatments. This study provides a novel understanding of the effects of 1,25OHD on osteogenic differentiation and mineralization of cMSCs depending on cell stage and maturity.

A study was conducted to understand the effects of 25-hydroxyvitamin D3 (25OHD) and 1,25-dihydroxyvitamin D3 (1,25OHD) administration on the expression of key genes related to osteogenesis, adipogenesis, myogenesis, and vitamin D3 metabolism in the chicken embryo. A total of 120 fertilized Cobb 500 eggs were used in the current study and were reared under standard incubation conditions. On embryonic day 3 (ED 3), PBS (C), PBS with 40ng 1,25OHD (1,25D-L), 200ng 1,25OHD (1,25D-H), 40ng 25OHD (25D-L), or 200ng 25OHD (25D-H) were injected into the dorsal vein of developing embryos. Whole embryos were harvested at 1, 3, and 6h post-injection for gene expression analyses (n=8). Gene expression for key osteogenesis markers (RUNX2: runt-related transcription factor 2; BMP2: bone morphogenetic protein 2; COL1A2: collagen type I alpha 2 chain; BGLAP: bone gamma-carboxyglutamate protein; SPP1: secreted phosphoprotein 1; and ALP: alkaline phosphatese), adipogenesis markers (PPAR-γ: peroxisome proliferator-activated receptor gamma; FASN: fatty acid synthase; and FABP4: fatty acid binding protein 4), myogenesis markers (MYOG: myogenin; MYOD1: myogenic differentiation 1; and MYF5: myogenic factor 5), and the enzyme responsible for vitamin D3 inactivation (CYP24A1: cytochrome P450 family 24 subfamily A member 1) were measured using real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). Data were normalized by the ΔΔCT method and analyzed using a one-way ANOVA. Results indicated that at 1h post-injection, no differences were found among treatments. At 3h, the early osteogenesis differentiation marker, ALP, was increased by 1,25D-H and 25D-H, and 25D-H also stimulated the expression of adipogenesis markers (FAPB4 and FASN). In contrast, the expression of myogenesis markers (MYOD1 and MYF5) was suppressed by 25OHD or 1,25OHD treatments, respectively. At 6h, a late osteogenic differentiation marker, SPP1, was increased by 25D-H. MYOD1 and MYF5 were continuously suppressed by 25OHD treatments or 1,25D-H. The evidence of vitamin D3 metabolite retention was assessed by measuring CYP24A1 expression. At 1h, there were no differences in CYP24A1 expression. At 3h, all treatments upregulated CYP24A1 expression relative to control (PBS) embryos. However, at 6h, only the 25D-H group retained higher CYP24A1 expression compared to the other treatments. In conclusion, the results suggested both 1,25OHD and 25OHD induced chicken embryo osteogenesis and adipogenesis, but inhibited myogenesis during early chicken embryo development. The higher dosage of 25OHD showed a possibility of a longer retention time in the embryos.

A study was conducted to understand the effects of 25-hydroxyvitamin D3 (25OHD) and 1,25-dihydroxyvitamin D3 (1,25OHD) administration on the expression of key genes related to osteogenesis, adipogenesis, myogenesis, and vitamin D3 metabolism in the chicken embryo. A total of 120 fertilized Cobb 500 eggs were used in the current study and were reared under standard incubation conditions. On embryonic day 3 (ED 3), PBS (C), PBS with 40ng 1,25OHD (1,25D-L), 200ng 1,25OHD (1,25D-H), 40ng 25OHD (25D-L), or 200ng 25OHD (25D-H) were injected into the dorsal vein of developing embryos. Whole embryos were harvested at 1, 3, and 6h post-injection for gene expression analyses (n=8). Gene expression for key osteogenesis markers (RUNX2: runt-related transcription factor 2; BMP2: bone morphogenetic protein 2; COL1A2: collagen type I alpha 2 chain; BGLAP: bone gamma-carboxyglutamate protein; SPP1: secreted phosphoprotein 1; and ALP: alkaline phosphatese), adipogenesis markers (PPAR-γ: peroxisome proliferator-activated receptor gamma; FASN: fatty acid synthase; and FABP4: fatty acid binding protein 4), myogenesis markers (MYOG: myogenin; MYOD1: myogenic differentiation 1; and MYF5: myogenic factor 5), and the enzyme responsible for vitamin D3 inactivation (CYP24A1: cytochrome P450 family 24 subfamily A member 1) were measured using real-time quantitative reverse transcription polymerase chain reaction (qRT-PCR). Data were normalized by the ΔΔCT method and analyzed using a one-way ANOVA. Results indicated that at 1h post-injection, no differences were found among treatments. At 3h, the early osteogenesis differentiation marker, ALP, was increased by 1,25D-H and 25D-H, and 25D-H also stimulated the expression of adipogenesis markers (FAPB4 and FASN). In contrast, the expression of myogenesis markers (MYOD1 and MYF5) was suppressed by 25OHD or 1,25OHD treatments, respectively. At 6h, a late osteogenic differentiation marker, SPP1, was increased by 25D-H. MYOD1 and MYF5 were continuously suppressed by 25OHD treatments or 1,25D-H. The evidence of vitamin D3 metabolite retention was assessed by measuring CYP24A1 expression. At 1h, there were no differences in CYP24A1 expression. At 3h, all treatments upregulated CYP24A1 expression relative to control (PBS) embryos. However, at 6h, only the 25D-H group retained higher CYP24A1 expression compared to the other treatments. In conclusion, the results suggested both 1,25OHD and 25OHD induced chicken embryo osteogenesis and adipogenesis, but inhibited myogenesis during early chicken embryo development. The higher dosage of 25OHD showed a possibility of a longer retention time in the embryos.

Bone health and body weight gain have significant economic and welfare importance in the poultry industry. Mesenchymal stem cells (MSCs) are common progenitors of different cell lineages such as osteoblasts, adipocytes, and myocytes. Specific oxysterols have shown to be pro-osteogenic and anti-adipogenic in mouse and human MSCs. To determine the effect of 20(S)-hydroxycholesterol (20S) on osteogenic, adipogenic, and myogenic differentiation in chicken, mesenchymal stem cells isolated from compact bones of broiler chickens (cBMSCs) were subjected to various doses of 20S, and markers of lineage-specific mRNA were analyzed using real-time PCR and cell cytochemistry. Further studies were conducted to evaluate the molecular mechanisms involved in lineage-specific differentiation pathways. Like human and mouse MSCs, 20S oxysterol expressed pro-osteogenic, pro-myogenic, and anti-adipogenic differentiation potential in cBMSCs. Moreover, 20(S)-Hydroxycholesterol induced markers of osteogenic genes and myogenic regulatory factors when exposed to cBMSCs treated with their specific medium. In contrast, 20S oxysterol suppressed expression of adipogenic marker genes when exposed to cBMSCs treated with OA, an adipogenic precursor of cBMSCs. To elucidate the molecular mechanism by which 20S exerts its differentiation potential in all three lineages, we focused on the hedgehog signaling pathway. The hedgehog inhibitor, cyclopamine, completely reversed the effect of 20S induced expression of osteogenic and anti-adipogenic mRNA. However, there was no change in the mRNA expression of myogenic genes. The results showed that 20S oxysterol promotes osteogenic and myogenic differentiation and decreases adipocyte differentiation of cBMSCs. This study also showed that the induction of osteogenesis and adipogenesis inhibition in cBMSCs by 20S is mediated through the hedgehog signaling mechanism. The results indicated that 20(S) could play an important role in the differentiation of chicken-derived MSCs and provided the theory basis on developing an intervention strategy to regulate skeletal, myogenic, and adipogenic differentiation in chicken, which will contribute to improving chicken bone health and meat quality. The current results provide the rationale for the further study of regulatory mechanisms of bioactive molecules on the differentiation of MSCs in chicken, which can help to address skeletal health problems in poultry.

Made available in DSpace on 2020-12-12T02:05:30Z (GMT). No. of bitstreams: 0 Previous issue date: 2020-04-28 An experiment was conducted to evaluate the effect of different levels of inorganic copper and zinc on growth performance, intestinal permeability, intestinal lesion scores, oocyst shedding, antioxidant properties and bone quality in broilers challenged with Eimeria spp. A total of 360 d-old male Cobb broiler chickens were housed in floor cages for 12 days at the Poultry Research Center. At 12 days of age, birds were placed in grower Petersime batteries and distributed in a completely randomized design with 10 birds per cage, six replicates per treatment, and six treatments. There were six corn-soybean meal-based dietary treatments: non-challenged control (NC), challenged control (CC), 100 ppm Cu (100 Cu), 150 ppm Cu (150 Cu), 80 ppm Zn (80 Zn), and 100 ppm Zn (100 Zn). Broilers received the treatment diets for 9 days (12–20d). Birds, except NC, were challenged with Eimeria maxima (50,000 oocysts/bird), Eimeria tenella (50,000 oocysts/bird), and Eimeria acervulina (250,000 oocysts/bird) on 14d. On 20d, the growth performance was recorded, and one bird/cage was used for analysis of intestinal permeability, antioxidant properties and bone quality. Lesion score was recorded at 20 days of age in eight birds/cage. The means were subjected to ANOVA and, when significant, compared by Duncan's test. Intestinal permeability was significantly improved when birds received the 100 Zn diet (P < 0.05). In addition, lesion scores on duodenum were reduced when broilers received diets 150 Cu as compared to CC diet (P < 0.05). However, growth performance was not positively influenced by inclusion of inorganic minerals as compared to the NC diet. Furthermore, activity of superoxide dismutase and bone quality were not affected, whereas glutathione status was improved with mineral supplementation in all groups. This study showed that Cu and Zn supplementation improves intestinal integrity during the Eimeria spp. infection, suggesting that Cu and Zn supplementation would be a potential strategy to reduce detrimental effects of Eimeria infection in broilers. Department of Breeding and Animal Nutrition School of Veterinary Medicine and Animal Science São Paulo State University (UNESP) Department of Poultry Science University of Georgia (UGA) Department of Foods and Nutrition University of Georgia (UGA) Department of Breeding and Animal Nutrition School of Veterinary Medicine and Animal Science São Paulo State University (UNESP)

Understanding of adipogenesis is important to find remedies for obesity and related disorders. In addition, it is also critical in bone disorders because there is a reciprocal relationship between adipogenesis and osteogenesis in bone micro-environment. Oxysterols are pro-osteogenic and anti-adipogenic molecules via hedgehog activation in pluripotent bone marrow stomal cells. However, no study has evaluated the role of specific oxysterols in C3H10T1/2 cells, which are a good cell model for studying osteogenesis and adipogenesis in bone-marrows. Thus, we investigated the effects of specific oxysterols on adipogenesis and expression of adipogenic transcripts in C3H10T1/2 cells. Treatment of cells with DMITro significantly induced mRNA expression of Ppar&gamma, This induction was significantly inhibited by 25-HC. The expression of C/cep&alpha, Fabp4 and Lpl was also inhibited by 25-HC. To determine the mechanism by which 25-HC inhibits adipogenesis, the effects of the hedgehog signalling pathway inhibitor, cyclopamine and CUR61414, were evaluated. Treatment of C3H10T1/2 cells with DMITro + cyclopamine or DMITro + CUR61414 for 96h did not modulate adipocyte differentiation, cyclopamine and CUR61414 did not reverse the inhibitory effects of 25-HC, suggesting that the canonical hedgehog signalling may not play a role in the anti-adipogenic effects of 25-HC in C3H10T1/2 cells. In addition, LXR agonist did not inhibit adipogenesis, but 25-HC strongly inhibits adipogenesis of C3H10T1/2 cells. Our observations showed that 25-HC was the most potent oxysterol in inhibiting adipogenesis and the expression of key adipogenic transcripts in C3H10T1/2 cells among the tested oxysterols, suggesting its potential application in providing an intervention in osteoporosis and obesity. We also report that the inhibitory effects of 25-HC on adipogenic differentiation in C3H10T1/2 cells are not mediated by hedgehog signaling and LXR.

This study was conducted to examine the effects of fatty acids (FA) with/without chicken serum (CS) on the expression of adipogenic transcripts and adipogenesis in chicken stromal vascular cells (SVC). In experiment 1, SVC were grown in DMEM containing 10% FBS (Control) and treated with 300 µM oleic acid (OLA) + FBS, linoleic acid (LNA) + FBS, palmitic acid (PAM) + FBS, or stearic acid (STA) + FBS for 48 h. In experiment 2, cells were grown in DMEM containing 5% CS and treated with 300 µM OLA (CS + OLA), PAM (CS + PAM), STA (CS + STA) or 200 µM LNA (CS + LNA) for 48 h. Adipogenesis was determined using Oil Red O staining and glycerol-3-phosphate dehydrogenase (GPDH) activity. The proportion of OLA, PAM, or STA was increased (P

A study was conducted to evaluate the effects of supplementing different levels of lysophospholipid (LPL) to normal or reduced energy diets on growth performance, carcass yield, intestinal morphology, and skeletal development in broilers. A total of 960 one-day-old Cobb 500 male birds were allocated using a 2 × 4 factorial arrangement with 2 energy levels (NE: normal and RE: 100 kcal/kg metabolizable energy reduction) and 4 LPL supplement levels (0, 0.025, 0.050, and 0.075%). Three diet phases were fed throughout the trial: starter (days 0 to 7), grower (days 8 to 21), and finisher (days 22 to 42) phases. Body weight (BW), feed intake (FI), and feed conversion ratio were calculated at the end of each phase. At day 7 and 21, duodenum and jejunum samples were collected for intestinal morphology and claudin-3 expression analyses, and tibia were sampled for bone quality analyses. At day 42, 4 birds per replicate were selected to measure carcass yield. The results showed low metabolizable energy diets impaired bird's growth performance, intestine development, and bone quality. The 0.075% LPL supplement in NE improved BW, BW gain, and FI in the finisher and overall period compared with no LPL supplement in NE (P < 0.05). In RE, the 0.025% LPL supplement significantly improved growth performance compared to the other treatments in RE (P < 0.05). The interactions on processing parameters were detected with LPL supplement in NE diets; 0.025, 0.05, and 0.075% LPL supplements significantly increased pectoral major percentages compared to the one without LPL supplement in NE (P < 0.05). The 0.075% LPL supplement increased dressing percentage (cold carcass weight/live BW) compared with the others (P < 0.05). The intestine morphology results showed LPL had positive effects on intestine development mainly during the early age (day 7) and claudin-3 expression at both day 7 and 21. Furthermore, LPL supplement significantly increased the total Ca and P deposition and positively affected the bone structure development. In summary, dietary LPL supplementation promoted growth performance, carcass yield, intestinal development, intestinal health, and bone quality.

1,25-dihydroxyvitamin D3 (1,25OHD) has been suggested to play an important role in osteogenic differentiation and mineralization. However, limited data have been reported in avian species. In the present study, the direct role of 1,25OHD on osteogenic differentiation and mineralization in chicken mesenchymal stem cells (cMSCs) derived from day-old broiler bones was investigated. cMSCs were treated with control media (C), osteogenesis media (OM), OM with 1, 5, 10, and 50 nM 1,25OHD, respectively. The messenger RNA (mRNA) samples were obtained at 24 and 48 h and 3 and 7 days to examine mRNA expression of key osteogenic genes [runt related transcription factor 2 (RUNX2), bone morphogenetic protein 2 (BMP2), collagen type I alpha 2 chain (COL1A2), bone gamma-carboxyglutamate protein (BGLAP), secreted phosphoprotein 1 (SPP1), and alkaline phosphatase (ALP)]. Cells were stained at 7, 14, and 21 days using Von Kossa (mineralization), Alizarin Red (AR; mineralization), and Alkaline Phosphatase (early marker) staining methods. From the mRNA expression results, we found a time-dependent manner of 1,25OHD on osteoblast differentiation and mineralization. In general, it showed an inhibitory effect on differentiation and mineralization during the early stage (24 and 48 h), and a stimulatory effect during the late cell stage (3 and 7 days). The staining showed 1,25OHD had an inhibitory effect on ALP enzyme activities and mineralization in a dosage-dependent manner up to 14 days. However, at 21 days, there was no difference between the treatments. This study provides a novel understanding of the effects of 1,25OHD on osteogenic differentiation and mineralization of cMSCs depending on cell stage and maturity.

Development of antibiotic resistance in foodborne pathogens, Salmonella spp. and Campylobacter, is a public health concern. Public demand to reduce the use of sub-therapeutic antibiotic growth promoters (AGP) in poultry feeding has resulted in greater adoption of antibiotic-free poultry production systems. There is a need to understand the effects of AGP removal from poultry feed on gut microbiota and its impact on prevalence of foodborne pathogens. The effect of antibiotic withdrawal from poultry feed on gut microbial community, host performance and immunity, and prevalence of Salmonella and Campylobacter was evaluated. Birds were raised on three phase diets (starter [d0-22], grower [d23-35] and finisher [d36-42]) with and without bacitracin dimethyl salicyclate (BMD). At early growth stage, bird performance was improved (P ≤ 0.05) with BMD treatment, whereas performance was better (P ≤ 0.05) in control group (no BMD in the feed) at the time of commercial processing. Acetate and butyrate production was affected (P ≤ 0.05) by age, whereas propionate production was affected (P ≤ 0.05) by both the treatment and age. The bacterial communities in the cecum were more diverse (P ≤ 0.001) and rich compared to the ileal communities, and they shifted in parallel to one another as the chicks matured. Differences in diversity and species richness were not observed (P > 0.05) between the BMD-fed and control groups. Comparing all ages, treatments and diets, the composition of cecal and ileal bacterial communities was different (P ≤ 0.001). Inclusion of BMD in the feed did not affect the bacterial phyla. However, predictable shift in the ileal and cecal bacterial population at lower taxonomic level was observed in control vs BMD-fed group. Cytokines gene expression (IL-10, IL-4, IFN-γ, beta-defensin, and TLR-4) was affected (P≤ 0.05) in the BMD-fed group at early stages of growth. The prevalence of foodborne pathogens, Campylobacter spp. and Salmonella spp. showed higher abundance in the ilea of BMD-fed chicks compared to control group. Overall, this study provided insight of the impact of AGP supplementation in the feed on gut microbial modulations, bird performance, host immunity and pathogen prevalence. This information can assist in designing alternative strategies to replace antibiotics in modern poultry production and for food safety.

The effects of Aluminum (Al) exposure on immune functions of cultured splenic T and B lymphocytes of rats were studied. The lymphocytes were isolated from spleen of healthy male Wistar rats weighing 110-120 g. The cultured cells in RPMI-1640 medium were exposed to 0 (control group), 0.035 (low-dose group), 0.07 (medial-dose group), and 0.14 (high-dose group) mg/mL Al(3+) as aluminum trichloride (AlCl(3)) in an incubator under 5% CO(2) at 37°C for 24 h. The T and B lymphocyte proliferation was measured with a tetrazolium dye colorimetric assay. The levels of interleukin (IL)-2, IL-6, and tumor necrosis factor (TNF)-α were determined by iodine [(125)I] IL-2, IL-6, and TNF-α radioimmunoassay kits, respectively. The proportions of CD3(+), CD4(+), and CD8(+) T lymphocytes were measured with a flow cytometer. The results showed that the T and B lymphocyte proliferation, the levels of IL-2, IL-6, TNF-α, the proportions of CD3(+) and CD4(+) T lymphocytes, and the ratio of CD4(+)/CD8(+) T lymphocytes were lowered by Al treatments, while the proportion of CD8(+) T lymphocytes was increased. These findings indicate that Al exposure can inhibit the immune functions of splenic T and B lymphocytes of rats in vitro.

The bioaccumulation and immunotoxicity of aluminum (Al) have been previously documented. Al accumulates in the organs of the organism, including spleen. Spleen is a peripheral organ of the immune system. The accumulated Al may alter the immune function. Here, we investigated the bioaccumulation of Al in spleen and its alterations in the immune system. Forty male Wistar rats (5 weeks old) weighed 110-120 g were orally exposed to aluminum trichloride (AlCl(3)) (0, 64.18, 128.36 and 256.72 mg/kg body weight) in drinking water for 120 days. The concentrations of spleen's Al, iron (Fe), copper (Cu), zinc (Zn), interleukin-2 (IL-2), tumor necrosis factor-α (TNF-α) and growth index were examined at the end of the experiment. The results showed that the concentrations of Al and Cu in the spleen were increased in an AlCl(3)-dose dependent manner, and the concentrations of spleen's growth index, Fe, Zn, IL-2 and TNF-α were reduced in AlCl(3)-treated rats. The results suggest that AlCl(3) can suppress the growth of spleen, disorder the balance of trace elements and inhibit the immune regulation of cytokines in the spleen. It indicates that AlCl(3) suppresses the immune function of spleen.