25-Hydroxycholesterol Inhibits Adipogenic Differentiation of C3H10T1/2 Pluripotent Stromal Cells.

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 Pparγ. This induction was significantly inhibited by 25-HC. The expression of C/cepα, 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.

Phenamil enhances the adipogenic differentiation of hen preadipocytes.

A study was conducted to examine the effect of phenamil on adipogenic differentiation and expression of key adipogenic transcripts in hen preadipocytes. Preadipocytes were isolated from 20-week old Single Comb White Leghorn hens (Gallas gallus, Lohman strain). The experiment lasted for 48 h and had six treatments. Non-treated control (C) cells, cells treated with dexamethasone, 3-isobutyl-1-methylxanthine, insulin, and oleic acid (DMIOA) (T1), DMIOA + 15 µM phenamil (T2), DMIOA + 30 µM phenamil (T3), 15 µM phenamil alone (T4), and 30 µM phenamil alone (T5). Neutral lipid accumulation and the mRNA expression of key adipogenic transcripts were measured in all treatments and compared. Lipid accumulation was detected in T1, T2, and T3 only. Expression of peroxisome proliferator receptor-activator gamma 2 (PPARγ2), the core enhancer binding protein α (C/EBPα), C/EBPß, fatty acid binding protein 4 (FABP4), and lipoprotein lipase (LPL) as well as ETS variant 4 (ETV4) and 5 was higher (P < 0.05) in T2, T3, T4, and T5 compared to C. Expression of these transcripts was higher (P < 0.05) in T2 and T3 compared to T4 and T5. The core enhancer binding protein α, C/EBPß, and FABP4 were highly expressed (P < 0.05) in T1 compared to C. However, the expression of PPARγ2, LPL, and ETV4 and ETV5 was not significantly different. Expression of C/EBPα, C/EBPß, and FABP4 was higher (P < 0.05) in T2 and T3 compared to T1. Expression of sterol regulatory element binding protein 1 (SREBP1) and leptin receptor (LEPR) was not significantly different among the treatments. In conclusion, phenamil enhances DMIOA-induced adipogenic differentiation of hen preadipocytes but does not induce adipogenesis by itself.

Bone sialoprotein keratan sulfate proteoglycan (BSP-KSPG) and FGF-23 are important physiological components of medullary bone.

Medullary bone is a specialized bone found in the marrow cavity of laying birds. It provides a significant contribution to the calcium supply for egg shell formation. Medullary bone is distinguished from cortical bone by the presence of large amounts of a keratan sulfate proteoglycan (KSPG). The aims of the present experiment are to confirm the identity of the core protein of KSPG, identify a marker of medullary bone metabolism, and determine whether changes in keratan sulfate (KS) concentration in blood are associated with the egg-laying cycle. Using two different isolation techniques- one specific for bone and another for blood- we have identified bone sialoprotein (BSP) to be the core protein of this KSPG. We also determined that the amount of keratan sulfate (KS) in laying hen blood fluctuates in synchrony with the egg-laying cycle, and thus can serve as a specific marker for medullary bone metabolism. During the course of this investigation, we also found FGF-23 (phosphatonin) to be expressed in medullary bone, in synchrony with the egg-laying cycle. Western blotting was used to demonstrate the presence of this peptide in both laying hen blood and medullary bone extracts. The importance of FGF-23 (phosphatonin) and parathyroid hormone in normalizing the dramatic changes in plasma calcium and phosphorus during the 24h egg-laying cycle is discussed.

Molecular Regulation of Adipogenesis and Potential Anti-Adipogenic Bioactive Molecules.

Adipogenesis is the process by which precursor stem cells differentiate into lipid laden adipocytes. Adipogenesis is regulated by a complex and highly orchestrated gene expression program. In mammalian cells, the peroxisome proliferator-activated receptor γ (PPARγ), and the CCAAT/enhancer binding proteins (C/EBPs) such as C/EBPα, ß and δ are considered the key early regulators of adipogenesis, while fatty acid binding protein 4 (FABP4), adiponectin, and fatty acid synthase (FAS) are responsible for the formation of mature adipocytes. Excess accumulation of lipids in the adipose tissue leads to obesity, which is associated with cardiovascular diseases, type II diabetes and other pathologies. Thus, investigating adipose tissue development and the underlying molecular mechanisms is vital to develop therapeutic agents capable of curbing the increasing incidence of obesity and related pathologies. In this review, we address the process of adipogenic differentiation, key transcription factors and proteins involved, adipogenic regulators and potential anti-adipogenic bioactive molecules.

Transcriptome analysis of hen preadipocytes treated with an adipogenic cocktail (DMIOA) with or without 20(S)-hydroxylcholesterol.

BACKGROUND: 20(S)-hydroxycholesterol (20(S)) potentially reduces adipogenesis in mammalian cells. The role of this oxysterol and molecular mechanisms underlying the adipogenesis of preadipocytes from laying hens have not been investigated. This study was conducted to 1. Analyze genes differentially expressed between preadipocytes treated with an adipogenic cocktail (DMIOA) containing 500 nM dexamethasone, 0.5 mM 3-isobutyl-1-methylxanthine, 20 µg/mL insulin and 300 µM oleic acid (OA) and control cells and 2. Analyze genes differentially expressed between preadipocytes treated with DMIOA and those treated with DMIOA + 20(S) using Affymetrix GeneChip® Chicken Genome Arrays. RESULTS: In experiment one, where we compared the gene expression profile of non-treated (control) cells with those treated with DMIOA, out of 1,221 differentially expressed genes, 755 were over-expressed in control cells, and 466 were over-expressed in cells treated with DMIOA. In experiment two, where we compared the gene expression profile of DMIOA treated cells with those treated with DMIOA+20(S), out of 212 differentially expressed genes, 90 were over-expressed in cells treated with DMIOA, and 122 were over-expressed in those treated with DMIOA+20(S). Genes over-expressed in control cells compared to those treated with DMIOA include those involved in cell-to-cell signaling and interaction (IL6, CNN2, ITGB3), cellular assembly and organization (BMP6, IGF1, ACTB), and cell cycle (CD4, 9, 38). Genes over-expressed in DMIOA compared to control cells include those involved in cellular development (ADAM22, ADAMTS9, FIGF), lipid metabolism (FABP3, 4 and 5), and molecular transport (MAP3K8, PDK4, AGTR1). Genes over-expressed in cells treated with DMIOA compared with those treated with DMIOA+20(S) include those involved in lipid metabolism (ENPP2, DHCR7, DHCR24), molecular transport (FADS2, SLC6A2, CD36), and vitamin and mineral metabolism (BCMO1, AACS, AR). Genes over-expressed in cells treated with DMIOA+20(S) compared with those treated with DMIOA include those involved in cellular growth and proliferation (CD44, CDK6, IL1B), cellular development (ADORA2B, ATP6VOD2, TNFAIP3), and cell-to-cell signaling and interaction (VCAM1, SPON2, VLDLR). CONCLUSION: We identified important adipogenic regulators and key pathways that would help to understand the molecular mechanism of the in vitro adipogenesis in laying hens and demonstrated that 20(S) is capable of suppressing DMIOA-induced adipogenesis.

The effect of dietary fructooligosaccharide supplementation on growth performance, intestinal morphology, and immune responses in broiler chickens challenged with Salmonella Enteritidis lipopolysaccharides.

This study was conducted to examine the effects of fructooligosaccharide (FOS) supplementation on growth performance, lymphoid organ weight, intestinal morphology, and immunological status in broilers (n=180) challenged with Salmonella Enteritidis lipopolysaccharides (LPS). Birds were randomly assigned into a 3×2 factorial arrangement that included 1) 3 dietary treatments from d one to 21: positive control (PC), wheat-corn-soybean meal based diet contained antibiotics (virginiamycin and monensin); negative control (NC), as PC without antibiotics; and NC+FOS, as NC supplemented with 0.5% FOS, and 2) 2 intraperitoneal injections: 2 mg/kg Salmonella Enteritidis LPS or sterile phosphate buffered saline (PBS) on d 21. Growth performance and relative lymphoid organ weight were not significantly different among the treatments. Villus height, crypt depth, and total mucosa thickness were significantly increased (P<0.05) in the ileum of broiler chickens fed NC+FOS when compared to PC and NC. Birds in NC+FOS treatment had reduced heterophil but increased monocyte count when compared to NC (P<0.05). Significant diet×challenge interaction was observed on natural IgY levels (P<0.0001), and a significant dietary effect was observed on specific IgY levels in chickens fed NC+FOS (P=0.003). Supplementation of FOS also increased the expression of interleukin (IL)-1ß, -10, and interferon (IFN)-γ mRNA in the ileum of the birds. In summary, Salmonella Enteritidis LPS challenge established significant differences in the immune responses in broiler chickens. FOS supplementation increased ileal mucosa thickness and elevated the expressions of certain cytokine genes. It also led to the alteration of leukocyte compositions and serum IgY levels in response to LPS challenge, suggesting FOS supplementation may be effective to induce protective outcomes in gut health and immunity of broiler chickens.

Effects of 25-(OH)D3 on fecal Ca and P excretion, bone mineralization, Ca and P transporter mRNA expression and performance in growing female pigs.

A study was conducted to examine the effects of 25-hydroxyvitamin D3 (25-(OH)D3) on fecal Ca and P excretion, bone mineralization, performance and the mRNA expression of intestinal transporter genes in growing female pigs. Sixty-day old gilts (n = 24) with an average initial BW of 23.13 ± 1.49 kg were randomly allocated to a control diet (diet 1) containing wheat/corn/soybean meal and 150 IU kg(-1) of Vitamin D3, diet 1 + 50 µg of 25-(OH)D3 kg(-1) (diet 2) and diet 1 + 100 µg of 25-(OH)D3 kg(-1) (diet 3). The pigs were housed in an individual pen and had ad libitum access to feed and water for 42 days, and BWG and feed intake were measured weekly. Measures of bone mineralization and expression of Ca and P transporters mRNA were analyzed using Dual Energy X-Ray Absortiometry (DEXA) and quantitative real-time polymerase chain reaction (qRT-PCR), respectively. Data were analyzed using GLM procedure of the Statistical Analysis System (SAS Institute version 9.2). Fecal Ca and P concentration were significantly reduced (P ≤ 0.05) in pigs fed diets 2 and 3 compared with the control diet. Supplementation of 25-(OH)D3 did not significantly improve bone mineralization, animal performance and intestinal transporters mRNA expression except for SLC34A1, a sodium-dependent phosphate transporter 1. In conclusion, supplementation of 25-(OH)D3 in swine nutrition may not improve animal performance but has the potential to reduce environmental pollution by increasing dietary Ca and P retention while reducing their excretion.

Entropically stabilized dislocations.

Dislocations are line defects that play a key role in the plasticity of crystalline materials and affect their thermal, chemical, and electrical properties. Typically dislocations are treated as stable defects; e.g., the equilibrium core structure of a dislocation is obtained by minimizing the crystal potential energy with respect to atom positions. Here we show for the first time the possibility of "entropically stabilized dislocations" that exist due to entropic effects without a corresponding potential energy well. An entropically stabilized dislocation was discovered in an accelerated multiscale quasicontinuum simulation. Its entropic nature was verified with fully atomistic free energy calculations and explained by a simple continuum-based model. This result has important consequences for the study of dislocations as well as for temporal multiscale methods that use information from the potential energy surface to accelerate time in molecular simulations.

A practical perspective on the implementation of hyperdynamics for accelerated simulation.

Consideration is given to several practical issues arising during the implementation of hyperdynamics, a methodology that extends the time scale of the conventional molecular dynamics simulation potentially by orders of magnitude. First, the methodology is reformulated in terms of the transition rate based on the buffer region approach (buffer rate), which can describe transitions in more general contexts than the transition state theory (TST). It will be shown that hyperdynamics can exactly preserve the buffer rate as well as the TST rate, which broadens the scope of the method. Next, the originally proposed scheme to compute the boost factor on-the-fly is reviewed and some alternative methods, one of which uses the umbrella sampling method, are presented. Finally, the methodology is validated in the context of a 1-dimensional example potential and a 3-dimensional simulation of the motion of an atomic force microscope tip moving along a surface.

Coccidiosis in Egg-Laying Hens and Potential Nutritional Strategies to Modulate Performance, Gut Health, and Immune Response.

Avian coccidiosis, despite advancements in management, nutrition, genetics, and immunology, still remains the most impactful disease, imposing substantial economic losses to the poultry industry. Coccidiosis may strike any avian species, and it may be mild to severe, depending on the pathogenicity of Eimeria spp. and the number of oocysts ingested by the bird. Unlike broilers, low emphasis has been given to laying hens. Coccidiosis in laying hens damages the gastrointestinal tract and causes physiological changes, including oxidative stress, immunosuppression, and inflammatory changes, leading to reduced feed intake and a drastic drop in egg production. Several countries around the world have large numbers of hens raised in cage-free/free-range facilities, and coccidiosis has already become one of the many problems that producers have to face in the future. However, limited research has been conducted on egg-laying hens, and our understanding of the physiological changes following coccidiosis in hens relies heavily on studies conducted on broilers. The aim of this review is to summarize the effect of coccidiosis in laying hens to an extent and correlate it with the physiological changes that occur in broilers following coccidiosis. Additionally, this review tries to explore the nutritional strategies successfully used in broilers to mitigate the negative effects of coccidiosis in improving the gut health and performance of broilers and if they can be used in laying hens.

Aflatoxin B1 Impairs Bone Mineralization in Broiler Chickens.

Aflatoxin B1 (AFB1), a ubiquitous mycotoxin in corn-based animal feed, particularly in tropical regions, impairs liver function, induces oxidative stress and disrupts cellular pathways, potentially worsening bone health in modern broilers. A 19-day experiment was conducted to investigate the effects of feeding increasing levels of AFB1-contaminated feed (<2, 75-80, 150, 230-260 and 520-560 ppb) on bone mineralization markers in broilers (n = 360). While growth performance remained unaffected up to Day 19, significant reductions in tibial bone ash content were observed at levels exceeding 260 ppb. Micro-computed tomography results showed that AFB1 levels at 560 ppb significantly decreased trabecular bone mineral content and density, with a tendency for reduced connectivity density in femur metaphysis. Moreover, AFB1 above 230 ppb reduced the bone volume and tissue volume of the cortical bone of femur. Even at levels above 75 ppb, AFB1 exposure significantly downregulated the jejunal mRNA expressions of the vitamin D receptor and calcium and phosphorus transporters. It can be concluded that AFB1 at levels higher than 230 ppb negatively affects bone health by impairing bone mineralization via disruption of the vitamin D receptor and calcium and phosphorus homeostasis, potentially contributing to bone health issues in broilers.

n-3 enriched Fish oil diet enhanced intestinal barrier integrity in broilers after Eimeria infection.

Coccidiosis caused by Eimeria spp. results in substantial economic losses in the poultry industry. The objective of this study was to investigate the effects of dietary supplementation with n-3 polyunsaturated fatty acids-enriched fish oil on growth performance, intestinal barrier integrity, and intestinal immune response of broilers challenged with Eimeria spp. A total of 576 fourteen-day-old broilers were randomly assigned in a completely randomized design with a 3 × 2 factorial arrangement, comprising 2 diets supplemented with either 5% fish oil or 5% soybean oil, and 3 Eimeria spp. infection levels: a nonchallenge control, a low dose of Eimeria challenge, and a high challenge dose. The results of the study revealed significant interactions between diet and Eimeria challenge to parameters of gut barrier integrity and feed intake. A significant interaction was observed in feed intake between 5 and 8 d postinfection (DPI), where the fish oil groups exhibited a higher amount of feed intake compared to the soybean oil diet groups after coccidiosis infection. The effects of the fish oil diet resulted in enhanced gut barrier integrity, as evidenced by a trend of decreased gastrointestinal leakage and a lower mean of small intestine lesion scores after Eimeria challenge. Additionally, significant interactions were noted between Eimeria spp. challenge and diet regarding jejunal crypt depth. The positive impact of the fish oil diet was particularly noticeable with the high Eimeria challenge dose. Overall, these findings underscore the relationship between the fish oil diet and Eimeria challenge on broiler chicken intestinal health. Dietary supplementation of fish oil has the potential to maintain small intestine barrier integrity with severe Eimeria infection conditions.

Water properties in intact wooden breast fillets during refrigerated storage.

The wooden breast (WB) condition notably alters moisture content and water holding capacity (WHC) in broiler breast fillets. The purpose of this study was to investigate water properties during refrigerated storage from 4 h to 168 h postmortem using time domain nuclear magnetic resonance (TD-NMR). Water properties measured included mobility (T), proportion (P), and abundance per 100 g of meat (A). Changes in meat quality indicators including compression force, color, pH, cumulative purge loss, and proximate composition were also measured. Compression force and energy of the WB fillets were higher than normal fillets (P < 0.05). Slopes of changes in lightness of the WB and normal fillets were different in skin and bone side (P < 0.05). The slope of the purge loss from the WB fillets was higher than the normal fillets (P < 0.05). Time domain nuclear magnetic resonance analysis showed 4 water populations in intact broiler fillets with transverse relaxation time (T2) constants at approximately 4 to 5 milliseconds (ms) (designated as 2b, corresponding to hydration water or bound water), 40 to 60 ms (designated as 21, corresponding to intra-myofibrillar water or immobilized water), 80 to 210 ms (designated as 22a, corresponding to extra-myofibrillar water or free water with lower mobility) and 210 to 500 ms (designated as 22b, corresponding to extra-myofibrillar water or free water with higher mobility) during early postmortem storage (between 4 h and 72 h postmortem) and only 3 populations (2b, 21, and 22a) after 72 h postmortem. There were interaction effects (P < 0.05) between storage time and WB condition for all water properties except T2b, A2b/100 g, and T22b. The linear change of T21, P21, A21/100 g, T22a, A22a/100 g, P22b, and A22b/100 g in stored WB samples were different from the normal fillets (P < 0.05). During storage, P21 and A21/100 g of the WB fillets exhibited faster linear increases than those of the normal fillets, whereas T21 and T22a of the normal fillets and A22a/100 g, P22b, and A22b/100 g of the WB fillets showed faster linear decreases (P < 0.05). Our data demonstrate that the WB condition affects changes in water properties in broiler fillets during postmortem refrigerated storage.

Effects of methionine supplementation in a reduced protein diet on growth performance, oxidative status, intestinal health, oocyst shedding, and methionine and folate metabolism in broilers under Eimeria challenge.

BACKGROUND: This study investigated effects of different methionine (Met) supplementation levels in a reduced protein diet on growth performance, intestinal health, and different physiological parameters in broilers under Eimeria challenge. A total of 600 fourteen-day-old Cobb500 male broilers were challenged with E. maxima, E. tenella, and E. acervulina, and randomly allocated in a 2 × 5 factorial arrangement. Birds received normal protein diets (20% crude protein, NCP) or reduced protein diets (17% crude protein, LCP), containing 2.8, 4.4, 6.0, 7.6, and 9.2 g/kg of Met. RESULTS: On 6 and 9 days post inoculation (DPI), increasing Met level linearly improved the growth performance (P < 0.05). Total oocyst shedding linearly increased as Met level increased (P < 0.05). Duodenal villus height (VH):crypt depth (CD) in the LCP groups were higher on 6 DPI (P < 0.01) while lower on 9 DPI (P < 0.05) compared to the NCP groups. Jejunal CD and duodenal VH:CD changed quadratically as Met level increased (P < 0.05). On 6 DPI, liver glutathione (GSH) and glutathione disulfide (GSSG) linearly increased as Met level increased (P < 0.05). On 9 DPI, GSSG quadratically increased, whereas GSH:GSSG quadratically decreased as Met levels increased (P < 0.05). The expression of amino acid transporters linearly decreased as Met level increased (P < 0.05). The expression of zonula occludens 2 and claudin-1 linearly increased on 6 DPI whereas decreased on 9 DPI as Met level increased (P < 0.05). The expressions of cytokines were lower in the LCP groups than the NCP groups (P < 0.05). Interaction effects were found for the expression of IL-10 and TNFα on 6 DPI (P < 0.05), where it only changed quadratically in the NCP group as Met level increased. The expression of Met and folate metabolism genes were lower in the LCP groups than the NCP groups on 9 DPI (P < 0.05). The expression of these genes linearly or quadratically decreased as Met level increased (P < 0.05). CONCLUSION: These results revealed the regulatory roles of Met in different physiological parameters including oxidative status, intestinal health, and nutrient metabolism in birds fed reduced protein diet and challenged with Eimeria.

Electrostatic particle ionization for suppressing air pollutants in cage-free layer facilities.

The increasing demand for cage-free (CF) poultry farming raises concern regarding air pollutant emissions in these housing systems. Previous studies have indicated that air pollutants such as particulate matter (PM) and ammonia (NH3) pose substantial risks to the health of birds and workers. This study aimed to evaluate the efficacy of electrostatic particle ionization (EPI) technology with different lengths of ion precipitators in reducing air pollutants and investigate the relationship between PM reduction and electricity consumption. Four identical CF rooms were utilized, each accommodating 175 hens of 77 wk of age (WOA). A Latin Square Design method was employed, with 4 treatment lengths: T1 = control (0 m), T2 = 12 ft (3.7 m), T3 = 24 ft (7.3 m), and T4 = 36 ft (11.0 m), where room and WOA are considered as blocking factors. Daily PM concentrations, temperature, and humidity measurements were conducted over 24 h, while NH3 levels, litter moisture content (LMC), and ventilation were measured twice a week in each treatment room. Statistical analysis involved ANOVA, and mean comparisons were performed using the Tukey HSD method with a significance level of P ≤ 0.05. This study found that the EPI system with longer wires reduced PM2.5 concentrations (P ≤ 0.01). Treatment T2, T3, and T4 led to reductions in PM2.5 by 12.1%, 19.3%, and 31.7%, respectively, and in small particle concentrations (particle size >0.5 µm) by 18.0%, 21.1%, and 32.4%, respectively. However, no significant differences were observed for PM10 and large particles (particle size >2.5 µm) (P < 0.10), though the data suggests potential reductions in PM10 (32.7%) and large particles (33.3%) by the T4 treatment. Similarly, there was no significant impact of treatment on NH3 reduction (P = 0.712), possibly due to low NH3 concentration (<2 ppm) and low LMC (<13%) among treatment rooms. Electricity consumption was significantly related to the length of the EPI system (P ≤ 0.01), with longer lengths leading to higher consumption rates. Overall, a longer-length EPI corona pipe is recommended for better air pollutant reduction in CF housing. Further research should focus on enhancing EPI technology, assessing cost-effectiveness, and exploring combinations with other PM reduction strategies.

Effects of phytase supplementation on broilers fed with calcium and phosphorus-reduced diets, challenged with Eimeria maxima and Eimeria acervulina: influence on growth performance, body composition, bone health, and intestinal integrity.

An experiment was conducted to evaluate the effects of phytase in calcium (Ca) and available phosphorous (avP)-reduced diet on growth performance, body composition, bone health, and intestinal integrity of broilers challenged with Eimeria maxima and Eimeria acervulina. A total of 672 14-day-old male broilers were allocated to a 2 × 4 factorial arrangement with 6 replicates per treatment and 14 birds per replicate. Two factors were Eimeria challenge and 4 dietary treatments: 1) a positive control (PC; 0.84% Ca and 0.42% avP); 2) a negative control (NC; 0.74% Ca and 0.27% avP); 3) NC + 500 FTU/Kg of phytase (NC + 500PHY); and 4) NC + 1,500 FTU/Kg of phytase (NC + 1500PHY). On d 14, birds in the Eimeria-challenged groups received a solution containing 15,000 sporulated oocysts of E. maxima and 75,000 sporulated oocysts of E. acervulina via oral gavage. At 5 d postinoculation (DPI), the challenged birds showed a higher (P < 0.01) FITC-d level than the unchallenged birds. While the permeability of the NC group did not differ from the PC group, the phytase supplementation groups (NC + 500PHY and NC + 1500PHY) showed lower (P < 0.05) serum FITC-d levels compared to the NC group. Interaction effects (P < 0.05) of Eimeria challenge and dietary treatments on feed intake (FI), mucin-2 (MUC2) gene expression, bone ash concentration, and mineral apposition rate (MAR) were observed. On 0 to 6 and 0 to 9 DPI, Eimeria challenge decreased (P < 0.01) body weight (BW), body weight gain (BWG), FI, bone mineral density (BMD), bone mineral content (BMC), bone area, fat free bone weight (FFBW), bone ash weight, bone ash percentage and bone ash concentration; and it showed a higher FCR (P < 0.01) compared to the unchallenged group. The reduction Ca and avP in the diet (NC) did not exert adverse effects on all parameters in birds, and supplementing phytase at levels of 500 or 1,500 FTU/Kg improved body composition, bone mineralization, and intestinal permeability, with the higher dose of 1,500 FTU/Kg showing more pronounced enhancements. There was an observed increase in FI (P < 0.01) when phytase was supplemented at 1,500 FTU/Kg during 0 to 6 DPI. In conclusion, results from the current study suggest that dietary nutrients, such as Ca and avP, can be moderately reduced with the supplementation of phytase, particularly in birds infected with Eimeria spp., which has the potential to save feed cost without compromising growth performance, bone health, and intestinal integrity of broilers.

Effects of Methionine Supplementation Levels in Normal or Reduced Protein Diets on the Body Composition and Femur Bone Characteristics of Broilers Challenged with Coccidia.

This study investigated the effects of dietary methionine (Met) levels on the bone quality of broilers challenged with coccidia. A total of 600 fourteen-day-old male Cobb500 broilers were gavaged with mixed Eimeria spp. and randomly allocated into 10 treatment groups by a 2 × 5 factorial arrangement. Birds received normal protein diets (NCP) or reduced-protein diets (LCP), containing 2.8, 4.4, 6.0, 7.6, and 9.2 g/kg of Met. Data were analyzed via two-way ANOVA and orthogonal polynomial contrast. At 9 days post-inoculation (DPI), whole body bone mineral density (BMD) and bone mineral content (BMC) linearly decreased as Met levels increased (p < 0.05). For the femoral metaphysis bone quality at 9 DPI, BMD linearly decreased, and porosity linearly increased as Met levels increased (p < 0.05) in the cortical bone. The increased Met levels linearly improved trabecular bone quality in LCP groups (p < 0.05) while not in NCP groups. For the femoral diaphysis cortical bone at 6 DPI, LCP groups had higher BMD and BMC than NCP groups (p < 0.05). Bone volume linearly increased as Met levels increased in LCP groups (p < 0.05) while not in NCP groups. In summary, the results suggested that increased Met levels decreased the cortical bone quality. However, in the context of reduced-protein diets, the increased Met levels improved trabecular bone quality.

Graded levels of Eimeria infection modulated gut physiology and temporarily ceased the egg production of laying hens at peak production.

An experiment was conducted to investigate the changes in gastrointestinal physiology, including intestinal leakage, immune response, oxidative stress, along with performance traits, of Hy-Line W-36 laying hens following Eimeria infection at peak egg production. A total of 360 laying hens, at 25 wk of age, were assigned randomly to 5 treatment groups, each consisting of 6 replicate cages, including a nonchallenged control group. The other 4 groups were inoculated with graded levels of mixed Eimeria species as high, medium-high, medium-low, and low doses, respectively. The body weight (BW) and body weight gain (BWG) of laying hens were measured from 0 to 14 days postinoculation (DPI). Average daily feed intake (ADFI) and hen-day egg production (HDEP) were measured from 0 to 15 and 0 to 28 DPI, respectively. Gut permeability was measured on 5 DPI, whereas oxidative stress, immune response, and expression of nutrient transporter genes were measured on 6 and 14 DPI. A significant linear reduction in BW and BWG was observed with increased Eimeria inoculation dosage on both 6 and 14 DPI (P < 0.001, P-Lin < 0.0001). An interaction between the Eimeria dosages and DPI was observed for ADFI (P < 0.0001). Feed intake in the challenged groups decreased starting at 4 DPI, with the most significant drop occurring at 7 DPI, which did not recover until 15 DPI. Following the challenge, gastrointestinal physiology shifted toward the host defense against the Eimeria infection by upregulating mRNA expression of tight junction proteins and immune responses while downregulating the expression of key nutrient transporters on 6 and 14 DPI (P < 0.05). An interaction between the Eimeria inoculation dosage and DPI was also observed for daily HDEP (P < 0.0001). Overall, HDEP was lower in the challenged groups compared to the control. Daily HDEP in the challenged groups dropped from 8 DPI and became similar to the control birds only after 24 DPI. Egg production temporarily ceased in most of the laying hens infected with the high and medium-high dosages of Eimeria. In conclusion, Eimeria infection activated the host immune response, negatively affecting the gastrointestinal health, growth performance, and temporarily ceased the egg production of Hy-Line W-36 laying hens when infected at peak production.

Comparison of necrotic enteritis effects on growth performance and intestinal health in two different meat-type chicken strains Athens Canadian Random Bred and Cobb 500.

Chickens have undergone genetic improvements in the past few decades to maximize growth efficiency. However, necrotic enteritis (NE), an enteric disease primarily caused by C. perfringens, remains a significant problem in poultry production. A study investigated the differences in intestinal health between the nonselected meat-type chicken Athens Canadian Random Bred (ACRB) and the modern meat-type Cobb 500 broilers (Cobb) when challenged with experimental NE. The study utilized a 2 × 3 factorial arrangement, consisting of two main effects of chicken strain and NE challenge model (nonchallenged control, NC; NE challenge with 2,500/12,500 Eimeria maxima oocysts + 1 × 109C. perfringens, NE2.5/NE12.5). A total of 432 fourteen-day-old male ACRB and Cobb were used until 22 d (8 d postinoculation with E. maxima on d 14, dpi), and the chickens were euthanized on 6 and 8 dpi for the analysis. All data were statistically analyzed using a two-way ANOVA, and Student's t-test or Tukey's HSD test was applied when P < 0.05. The NE12.5 group showed significant decreases in growth performance and relative growth performance from d 14 to 20, regardless of chicken strain (P < 0.01). The ACRB group exhibited significant decreases in relative body weight and relative body weight gain compared to the Cobb group from d 14 to 22 (P < 0.01). On 6 and 8 dpi, both NE challenge groups showed significant decreases in intestinal villus height to crypt depth ratio, jejunal goblet cell count, and jejunal MUC2 and LEAP2 expression (P < 0.01). Additionally, the NE12.5 group had significantly higher intestinal NE lesion score, intestinal permeability, fecal E. maxima oocyst count, intestinal C. perfringens count, and jejunal IFNγ and CCL4 expression compared to the NC group (P < 0.05). In conclusion, NE negatively impacts growth performance and intestinal health in broilers, parameters regardless of the strain.

Effects of Artemisia annua supplementation on the performance and gut health of laying hens challenged with mixed Eimeria species.

Background: Coccidiosis outbreaks in susceptible laying hens can significantly decrease egg production and cause substantial economic loss to the egg industry. The supplementation of poultry diets with chemotherapeutic agents is limited due to antimicrobial resistance and residue in poultry meat or processed products. Therefore, alternative strategies to control coccidiosis are needed, and Artemisia annua (AA) might have the potential to be a phytogenic feed additive, an alternative to anticoccidial agents. This study aimed to investigate the effect of the dietary supplementation of powdered AA on the performance and gut health of laying hens infected with coccidiosis by Eimeria spp. Methods: A total of 225 Hy-Line W-36 laying hens at 23 weeks of age were allocated into 5 treatment groups: 1) control (NC), 2) pair-fed (PF) control, 3) challenged control (CC), 4) CC with dietary inclusion of 0.5% AA (0.5AA), and v) CC with dietary inclusion of 1% AA (1AA). The hens in the CC, 0.5AA, and 1AA groups were orally inoculated with sporulated oocysts of Eimeria maxima (12,500), Eimeria tenella (12,500), and Eimeria acervulina (62,500) at week 25. The PF hens received the same amount of feed consumed by the CC hens from 0-14 days post-inoculation (dpi) of Eimeria spp. The performance of the laying hens, including body weight (BW), hen-day egg production (HDEP), feed intake (FI), and feed conversion ratio (FCR), was measured weekly. Additionally, markers of intestinal health, including gut permeability, lesion score, intestinal morphometry, and immune responses, were evaluated at 6, 14, and 21 dpi. Results: At 6 and 14 dpi, laying hens challenged with Eimeria spp. had a lower BW than PF and NC hens (p < 0.0001). Supplementation of 1% AA improved the HDEP by 8.1% compared to CC hens; however, it was still 15.4% lower than that of PF hens (p < 0.0001). The inclusion of 1% AA did not have any beneficial effect on FI; however, the FCR was improved by 0.61 (2.46) than that of CC hens (3.07; p < 0.0001). The inclusion of 1% AA reduced the severity of the intestinal lesions and increased the recovery of intestinal villi (p < 0.05). Additionally, gut permeability was significantly different between the challenged and non-challenged hens; however, among the challenged hens, the inclusion of AA reduced the gut permeability by 29% compared to CC hens (p < 0.0001). Furthermore, the inclusion of 0.5% AA reduced the inflammatory responses in the infected hens. Conclusion: Dietary inclusion of AA partially restored the performance and gut health of the laying hens and modulated their inflammatory immune response following Eimeria infection; however, further studies are needed to better understand the mode of action and effective dosages to improve the gut health without negative impacts on the performance.