Research Note: Effect of dietary xylo-oligosaccharide on growth performance, intestinal histomorphology, and specific cecal bacteria in broiler chickens.

This study was conducted to evaluate the prebiotic effect of xylo-oligosaccharide (XOS) supplemented in a corn-soybean meal (SBM) based conventional diet on growth performance, intestinal histomorphology, and quantification of specific bacteria in the ceca of broilers. A total of 240 d of hatch Cobb 500 male broiler chicks were randomly assigned to 4 dietary treatments (corn-SBM-based control diet) containing 0, 0.05, 0.1, and 0.2% XOS. The broilers were raised for 21 d in 6 replicate cages, each containing 10 birds. Growth performance parameters were obtained weekly. Additionally, small intestinal tissues were collected to evaluate histomorphometry and whole ceca were collected to quantify bacterial populations on D21. The results showed that inclusion of XOS has similar body weight (BW), body weight gain (BWG), feed intake (FI), and feed conversion ratio (FCR) as the control group during the 21-day study. The results further indicate a tendency for the jejunum villus to crypt ratio (VH:CD) to increase in birds given 0.05 and 0.2% XOS (P = 0.08). Cecal bacteria quantification showed a linear increase in Bifidobacterium with increasing XOS levels (P < 0.0001) and a decrease Clostridium perfringens levels compared to birds fed the control diet (P < 0.0001). However, there were no differences in the total counts of Lactobacillus and E. coli. Together these results showed that while there were no differences in growth parameters up to 21 d, the histomorphology findings and the increase in Bifidobacterium, along with the reduction in C. perfringens observed in the XOS groups, suggest a beneficial impact of XOS inclusion on gut health. Further research with longer feeding periods and higher XOS levels should be conducted to explore potential positive effects on both growth and gut health parameters.

Temporal dynamics of innate and adaptive immune responses in broiler birds to oral delivered chitosan nanoparticle-based Salmonella subunit antigens.

Salmonella enterica serovar Enteritidis (S. Enteritidis, SE) infection of poultry causes a significant risk to public health through contamination of meat and eggs. Current Salmonella vaccines have failed to provide strong mucosal immunity in the intestines to reduce Salmonella shedding and food contamination. Considering the short lifespan of broilers, an easy-to-deliver, safe and effective Salmonella vaccine is urgently needed. Our goal in this study was to demonstrate the ability of chitosan nanoparticle (CNP) vaccine delivery platform in activating immune response to Salmonella antigens in broilers inoculated orally. In an initial study, soluble whole antigen of SE entrapped in CNP was inoculated but the specific immune responses were poor. Therefore, the CNP entrapped immunogenic outer membrane proteins (OMP) and flagellin (FLA) of SE and surface conjugated with FLA [CNP-(OMP + FLA)] was developed. In broilers inoculated orally with CNP-(OMP + FLA) formulation once or twice, we monitored the temporal expression of innate immune molecules and antigen specific lymphocyte proliferation. In the cecal tonsils of CNP-(OMP + FLA) inoculated birds, we observed enhanced expression of mRNA coding Toll-like receptors (TLRs)- 1, 4, 5, and 7, especially at dpv 21. In addition, both OMP and FLA specific lymphocytes proliferation at dpv 7 and 21 by CNP-(OMP + FLA) were enhanced in the spleen. In conclusion, CNP-(OMP + FLA) formulation augmented both innate and lymphocyte responses in orally inoculated broilers. Further studies are needed to determine the candidate subunit CNP vaccine's efficacy in a challenge trial.

Research Note: Effect of synbiotic supplementation on caecal Clostridium perfringens load in broiler chickens with different necrotic enteritis challenge models.

Studies were conducted to determine the efficacy of synbiotic applications to combat the negative effects of necrotic enteritis (NE). An in vitro study was conducted to test the effect of probiotics species supernatants to decrease Clostridium perfringens (CP) proliferation. Lactobacillus reuteri, Enterococcus faecium, Bifidobacterium animalis, and Pediococcus acidilactici culture supernatants decreased the proliferation of CP at 1:1 supernatant-to-pathogen dilution in vitro. Two in vivo studies were conducted to determine the in vivo response of synbiotic supplementation containing the aforementioned probiotic strains on broiler production performance and caecal CP load in broilers induced with NE infection. In experiment 1, 75 broiler chicks were randomly allotted to 3 treatment groups, control (basal diet), ionophore (Salinomycin), and synbiotic (PoultryStar me), from day of hatch, and NE was induced in all birds. There were no significant treatment effects on BW, feed consumption, and feed gain ratio. However, at 35 D, ionophore or synbiotic supplementation increased (P < 0.05) villi height and decreased interleukin (IL)-1 mRNA abundance, while synbiotic supplementation increased (P < 0.05) IL-10 mRNA abundance compared with the control group, respectively. In experiment 2, 360 broiler chicks were randomly allotted to 3 treatments, an unchallenged negative control (control; basal diet), challenged positive control (NE; basal diet), or NE + synbiotic group (synbiotic). At both 21 and 42 D of age, NE birds had decreased (P < 0.05) BW, feed conversion, and jejunal villi height compared with control, while NE + synbiotic birds were not different from control groups. At 42 D of age, NE birds had 2.2 log/g increased CP in the ceca contents compared with control, while synbiotic birds had CP load that was not different than that of the control group. NE + synbiotic birds had significantly greater amounts of bile anti-CP IgA than the control and NE groups. It can be concluded that synbiotic supplementation decreased CP proliferation in vitro and caecal CP load in vivo while improving production parameters during an NE infection in broilers.

Synbiotic supplementation to decrease Salmonella colonization in the intestine and carcass contamination in broiler birds.

In vitro and in vivo experiments were conducted to study the effects of synbiotic supplementation on Salmonella enterica ser. Enteritidis (SE) proliferation, cecal content load, and broiler carcass contamination. Lactobacillus reuteri, Enterococcus faecium, Bifidobacterium animalis, and Pediococcus acidilactici culture supernatants decreased (P < 0.05) the in vitro proliferation of SE at 1:1 supernatant: pathogen dilution. A total of 240 Cobb-500 broiler chicks were randomly allotted to three treatment groups (8 replicates/group with 10 birds/replicate): control (basal diet), antibiotic (Virginiamycin at 20 mg/kg feed), synbiotic (PoultryStar® ME at 0.5 g/kg feed containing L. reuteri, E. faecium, B. animalis, P. acidilactici and a Fructooligosaccharide) from day of hatch. At 21 d of age, all birds in experimental groups were orally inoculated with 250 µl of 1 X 109 CFU SE. Antibiotic supplementation increased (P < 0.05) body weight and feed consumption, compared to the control group. Birds in the synbiotic supplementation had intermediate body weight and feed consumption that were not significantly different from both the control and antibiotic group at 42 d of age in SE infected birds. No significant effects were observed in feed efficiency at 42 d of age among the groups. Antibiotic and synbiotic supplementation decreased (P < 0.05) SE load in cecal contents by 0.90 and 0.85 log units/ g and carcass SE load by 1.4 and 1.5 log units/mL of rinsate compared to the control group at 42 d of age (21 dpi). The relative abundance of IL-10, IL-1, TLR-4, and IFNγ mRNA was decreased (P < 0.05) in the antibiotic and synbiotic supplementation groups compared to the control birds at 42 d of age (21 dpi). It can be concluded that synbiotic supplementation decreased SE proliferation in vitro and decreased SE load in the cecal contents and broiler carcass.

Effects of hydroxychloride and sulfate form of zinc and manganese supplementation on superoxide dismutase activity and immune responses post lipopolysaccharide challenge in poultry fed marginally lower doses of zinc and manganese.

Effects of hydroxychloride (OHCl) and sulfate form of zinc and manganese supplementation on immune responses of birds fed marginally lower levels of zinc and manganese during an experimental lipopolysaccharide (LPS) injection were studied. In experiment I, 30-week-old layer birds were fed 50 mg/kg Zn+45 mg/kg Mn or 100 mg/kg Zn+90 mg/kg Mn in sulfate or OHCl form and injected with 0 or 500 µg/kg LPS in a 2 (50 mg Zn+45 mg Mn and 100 mg Zn+90 mg Mn) X 2 (sulfate and OHCl) X 2 (0 and 500 µg LPS) factorial setup of treatments for 10 weeks. Among LPS-injected birds, those receiving 50 mg ZnOHCl+45 mg MnOHCl had comparable heterophil and monocyte superoxide dismutase (SOD) activity compared to the birds fed 100 mg Zn+90 mg Mn. Compared to the birds injected with PBS, LPS injection upregulated cathelicidin and IL-1 relative mRNA amounts in monocytes from birds fed 100 mg Zn+90 mg Mn, both in sulfate and OHCl form, and in birds fed 50 mg ZnOHCl+45 mg MnOHCl, but not in the birds fed 50 mg ZnSO4+45 mg MnSO4. In experiment II, one-day-old broiler birds were fed 50 mg ZnOHCl+45 mg MnOHCl, 50 mg ZnOHCl+90 mg MnOHCl, 100 mg ZnOHCL+45 mg MnOHCl, 100 mg ZnOHCl+90 mg MnOHCl, 50 mg ZnSO4+45 mg MnSO4, or 100 mg ZnSO4+90 mg MnSO4 for 21 and 42 days. Birds fed 100 mg ZnOHCl+45 mg MnOHCl form had a comparable heterophil and monocyte SOD activity and monocyte cathelicidin mRNA amounts compared to the group fed 100 mg Zn+90 mg Mn. Increasing the ZnOHCl content from 50 mg to 100 mg/kg Zn reversed (P > 0.05) the decrease in SOD activity and monocyte cathelicidin mRNA levels of the 50 mg ZnOHCl+45 mg MnOHCL fed group, and increasing the MnOHCl content from 45 mg to 90 mg/kg in the 100 mg ZnOHCl+45 mg MnOHCl group further increased SOD activity. In conclusion, birds fed diets with lower amounts of zinc and manganese in sulfate form decreased SOD activity and IL-1 and cathelicidin amounts during inflammation, and either increasing the dietary zinc and manganese content or feeding zinc and manganese in OHCl form synergistically increased the SOD activity and IL-1 and cathelicidin mRNA amounts in immune cells.

Effect of in vitro and in vivo 25-hydroxyvitamin D treatment on macrophages, T cells, and layer chickens during a coccidia challenge.

This article describes the in vitro and in vivo effects of a 25-hydroxycholecalciferol (25[OH]D) treatment in layer hens during a mixed coccidia challenge. HD11 cells (chicken macrophage cell line) were treated in vitro with a coccidia antigen or in a medium supplemented with either 1,25-dihydroxycholecalciferol (1,25[OH]2D) or 25(OH)D. HD11 cells treated in vitro with 200 nM of 1,25(OH)2D had increased nitrite production (P < 0.01) compared with HD11 cells treated with 0 or 200 nM of 25(OH)D. Treating HD11 cells with 25(OH)D decreased IL-10 mRNA by 1.7-fold, but 1,25(OH)2D treatment increased the amount of IL-10 mRNA by 2.7-fold (P < 0.01) compared with the group treated with 0 nM of 25(OH)D. Post-coccidial antigen stimulation, 25(OH)D or 1,25(OH)2D treatment decreased (P < 0.01) 1α-hydroxylase mRNA amounts in HD11 cells. Stimulating primary T cells in vitro with Concanavalin A (Con-A) decreased (P = 0.020) the 1α-hydroxylase mRNA amounts by 3-fold. ConA-B1-VICK cells (chicken T cell line) stimulated with 100 nM 1,25(OH)2D or with supernatants from HD11 cells treated with 25(OH)D plus lipopolysaccharide (LPS) had 1.3-fold less (P < 0.01) interferon (IFN)-γ mRNA compared with the group treated with 25(OH)D. Layer birds were fed a basal diet supplemented with 25(OH)D at 6.25, 25, 50, or 100 µg/kg, and at 21 d of age orally challenged with 1 × 10(5) live coccidia oocysts. Compared with birds fed similar levels of 25(OH)D and unchallenged with the coccidia oocyst, birds challenged with the coccidia oocyst had 15% reduced BW gain in the groups supplemented with either 6.25, 25, or 50 µg/kg of 25(OH)D, but only a 4% reduced BW gain in birds fed 100 µg/kg of 25(OH)D (P < 0.01). Birds fed 100 µg/kg 25(OH)D had decreased (P = 0.012) CD8+ cell percentages in cecal tonsils in both coccidial oocyst challenged and unchallenged birds, compared with birds fed 6.25 µg/kg 25(OH) and unchallenged with coccidial oocysts. At 15 d post-coccidia challenge, birds fed 100 µg/kg 25(OH)D and challenged with coccidial oocysts had 17% more CD4+CD25+ cells (P = 0.018) in the cecal tonsil compared with the birds fed 100 µg/kg 25(OH)D and unchallenged with coccidial oocysts. At d 6 post-coccidia challenge, birds fed 100 µg/kg 25(OH)D had a 3.5-fold increase (P < 0.01) in IL-10 mRNA amounts in the cecal tonsils compared with birds fed 6.25 µg/kg 25(OH)D. In conclusion, supplementing birds with 100 µg/kg 25(OH)D could be a nutritional strategy to reduce the production losses post-coccidia challenge.

Vitamin D-1α-hydroxylase and vitamin D-24-hydroxylase mRNA studies in chickens.

A series of experiments were conducted to study the basal amounts of vitamin D-1α-hydroxylase and vitamin D-24-hydroxylase mRNA amounts in different organs and the effect of immune stimulation on 1α- and 24-hydroxylase mRNA amounts in chickens. At day of hatch, kidneys had an approximately 66-fold higher amount of 1α-hydroxylase and 550-fold higher amount of 24-hydroxylase mRNA, thigh and breast muscles had an approximately 20-fold higher amount of 1α-hydroxylase mRNA, and the thymus had an approximately 41-fold higher amount of 24-hydroxylase mRNA than the liver. An in vivo LPS injection did not alter the amount of 1α-hydroxylase mRNA in the breast muscle (P=0.60) or in the kidneys (P=0.39). An in vivo LPS injection decreased (P=0.01) the amount of 24-hydroxylase mRNA in the breast muscle at 3 d post-LPS injection. An in vivo LPS injection increased (P=0.01) the amount of 24-hydroxylase mRNA in the kidneys at 2, 3, and 6 d post-LPS injection. An in vitro stimulation altered amounts of 1α- (P=0.01) and 24-hydroxylase (P=0.04) mRNA in CD4+ cells. In conclusion, the distribution of 1α- and 24-hydroxylase mRNA amounts was similar to mammals, and an immune stimulation altered the amounts of 1α- and 24-hydroxylase mRNA in chickens.

Regulatory T cell properties of thymic CD4+CD25+ cells in turkeys.

Thymic CD4+CD25+ cells from turkeys were analyzed for T regulatory cell suppressive properties and cytokine production profile. Thymocytes treated with concanavalin-A and interleukin-2 upregulated CD25 at 72 h of culture. The mean CD4+CD25+ cell percentage in the blood was 1.1%, in the thymus was 2.7%, in the spleen was 9.6%, in the lung was 18.6%, in cecal tonsils was 9.2%, and in bone marrow was 14.9%. The CD4+CD25+ cells were not detectable in the bursa. Turkey thymic CD4+CD25+ cells had approximately 158-fold higher interleukin-10 mRNA, 7-fold higher transforming growth factor ß, 24-fold higher cytotoxic T lymphocyte antigen 4, and 11-fold higher lymphocyte-activation gene 3 mRNA amounts than thymic CD4+CD25- cells. Thymic CD4+CD25+ cells had no detectable levels of interleukin-2 mRNA. Turkey CD4+CD25+ cells suppressed naïve cell proliferation in vitro. It could be concluded that thymic CD4+CD25+ cells in turkeys are likely the counterpart of mammalian T regulatory cells.

Effects of in ovo interleukin-4-plasmid injection on anticoccidia immune response in a coccidia infection model of chickens.

Two experiments were conducted to study the effects of an in ovo interleukin (IL)-4 plasmid injection in a coccidia infection model. In experiment I, chicks were hatched from eggs that had been injected in ovo with an empty vector or with 10 or 15 µg of IL-4 plasmid, and then challenged posthatch with coccidia. In experiment II, chicks were hatched from eggs that had been vaccinated with coccidia and injected in ovo with an empty vector or with 10 or 15 µg of IL-4 plasmid, and then challenged posthatch with coccidia. In experiment II, the BW gain of birds hatched from eggs vaccinated with live oocysts plus 15 µg of IL-4 plasmid was 25% higher than the BW gain of birds hatched from eggs vaccinated with live oocysts plus empty plasmid. In both experiments I and II, a 15-µg IL-4-plasmid injection decreased fecal oocyst shedding, decreased the number of CD8(+) cells in the cecal tonsils, and decreased cecal tonsil lymphocyte cell proliferation postcoccidia challenge. In experiment I, splenic macrophages of chicks hatched from eggs injected with 15 µg of IL-4 plasmid had higher nitric oxide production than those of chicks hatched from eggs injected with the empty plasmid. In experiment II, a 15-µg IL-4-plasmid injection increased serum anticoccidia IgG postcoccidia challenge. It could be concluded that 15 µg of IL-4 plasmid improved anticoccidia immune responses synergistically with in ovo coccidia vaccination in chickens.

In vivo lipopolysaccharide injection alters CD4+CD25+ cell properties in chickens.

In chickens, thymic CD4(+)CD25(+) cells are characterized as regulatory T cells. The objectives of this experiment were to study the effects of an in vivo lipopolysaccharide (LPS) injection on the percentage of CD4(+)CD25(+) cells in peripheral organs and the suppressive properties of splenic CD4(+)CD25(+) cells in chickens. Chickens were injected with LPS and CD4(+)CD25(+) cells were analyzed at 1, 2, 3, 5, and 10 d post LPS injection. The LPS injection increased CD4(+)CD25(+) cell percentage approximately 5-fold in the blood at 1 d post LPS injection (P < 0.001), 3-fold in the thymus at 3 d post LPS injection (P = 0.001), and 2.5-fold in the spleen at 2 d post LPS injection (P = 0.001) compared with the no-LPS-injected group. The LPS injection did not alter the CD4(+)CD25(+) cell percentage in the cecal tonsil (P = 0.162), lung (P = 0.098), or bone marrow (P = 0.071) at any time point measured. At 2 d post LPS injection, splenic CD4(+)CD25(+) cells lost their suppressive ability (P < 0.001). At 5 d post LPS injection, splenic CD4(+)CD25(+) cells not only regained their suppressive ability, but also became supersuppressive (P < 0.001). Splenic CD4(+)CD25(+) cells at 5 d post LPS injection produced 5.5-fold more (P = 0.005) IL-10 mRNA than splenic CD4(+)CD25(+) cells at 0 and 2 d post LPS injection. In conclusion, chicken regulatory T cells are differentially activated to facilitate immune response during the early stage of inflammation and to facilitate immune suppression at a later stage of inflammation.

Effect of recycled litter on immune cells in the cecal tonsils of chickens.

This experiment was conducted to study selected aspects of the gut immune response in broiler chicks reared on fresh or recycled litter that were fed diets with and without subtherapeutic antibiotic supplementation. All of the chicks were reared in pens that contained either fresh pine shavings (fresh litter) or litter that was recycled for 3 consecutive flocks (recycled litter). The experiment was a 2 × 2 factorial arrangement of treatments with 4 replicate pens (n = 4) per treatment. At 10 and 35 d of age, the cecal tonsils were analyzed for intestinal immune measurements. The cecal tonsils of birds reared on recycled litter had increased IL-1 mRNA (P < 0.01) and a lower percentage of CD4(+)CD25(+) cells at 10 and 35 d of age when compared with those of chicks reared on fresh litter. Birds fed diets supplemented with bacitracin had a reduction in CD4(+) cells (P = 0.01) at 10 d of age when compared with that of chicks that were not fed the antibiotic. The combination of bacitracin supplementation and fresh litter resulted in an approximate 10-fold increase in IL-10 mRNA (P = 0.01) at 10 d of age when compared with that of the unsupplemented chicks in fresh litter. Among those chicks that were not supplemented with bacitracin, the recycled-litter treatment resulted in 25-fold (P = 0.01) and 39-fold (P = 0.02) higher IL-4 mRNA levels at 10 and 35 d of age, respectively, when compared with those of the chicks reared on fresh litter. In conclusion, the intestinal immune response of birds reared on recycled litter is skewed toward an inflammatory response, whereas the fresh litter treatment was skewed toward an anti-inflammatory response. Bacitracin supplementation did not interact with the litter type to alter IL-1 mRNA levels in cecal tonsils, suggesting the low efficiency of bacitracin in alleviating the inflammatory response induced by recycled litter.

Effect of killed whole yeast cell prebiotic supplementation on broiler performance and intestinal immune cell parameters.

Two experiments were conducted to study the effect of CitriStim, a commercial killed whole yeast cell prebiotic, on broiler performance, regulatory T cells, CD4(+) and CD8(+) percentages, and IL-10 and IL-1 mRNA contents of the spleen and cecal tonsils. No immune challenges were imposed in either of the 2 experiments. One-day-old broiler chicks were fed a corn- and soybean meal-based diet supplemented with 0, 0.1, or 0.2% CitriStim (ADM, Decatur, IL) for 35 d. At 21 (P = 0.03) and 35 d (P = 0.02) of age, CitriStim supplementation at 0.2% increased regulatory T cell percentage in the cecal tonsil compared with that of the 0% CitriStim-supplemented group. At 21 (P = 0.08) and 35 d (P = 0.01) of age, CitriStim supplementation at 0.2% increased IL-10 mRNA content of the cecal tonsil compared with that of the 0% CitriStim-supplemented group. At 21 (P = 0.13) and 35 d (P < 0.01) of age, CitriStim supplementation at 0.2% decreased IL-1 mRNA content compared with that of the 0% CitriStim supplemented group. CitriStim supplementation did not (P > 0.05) alter the IL-10 and IL-1 mRNA contents in the spleen. CitriStim supplementation did not (P > 0.05) alter the CD4(+) and CD8(+) cell percentages in the spleen and cecal tonsil at 21 and 35 d of the experiment. CitriStim supplementation increased regulatory T cell percentage and IL-10 mRNA content and decreased IL-1 mRNA content in the cecal tonsil to produce a net antiinflammatory milieu. The immunomodulatory effect of CitriStim supplementation was a local effect rather than a systemic effect.

Intestinal physiology and regulatory T cell response to immediate or delayed access to feed and water in Pekin ducklings.

This study investigated the effects of delayed access to feed and water on early duckling growth, selected aspects of intestinal physiology, and immune responses. Ducklings were assigned to one of 2 experimental groups. In one treatment group (referred to as the fed group), feed and water were provided beginning on d 0, and in the other treatment group (referred to as the withheld group), feed and water were withheld until d 2. The ducklings in the withheld treatment had lower BW at 2 and 6 d posthatch than ducklings in the fed group. At 8 d posthatch (6 d postfeeding), the BW of the ducklings in the withheld group was comparable to the BW of the 6-d-old ducklings in the fed group. At 2 d posthatch, withheld ducklings had lower absolute and relative duodenum plus pancreas weight than fed ducklings. At 8 d posthatch (6 d postfeeding), withheld ducklings had higher absolute and relative duodenum plus pancreas weight than fed ducklings at 6 d posthatch. At 2 d posthatch, mucin 5B mRNA content was approximately 2 times lower in withheld ducklings (P = 0.09) than in fed ducklings. At 6 d posthatch, mucin 5b mRNA content was approximately 2.5-fold higher in withheld ducklings (P = 0.07) than in fed ducklings. Delayed access to feed and water increased the CD25(+) cell number in the cecal tonsil at 2, 6, and 8 d posthatch. The IL-10 content of CD25(+) cells was higher in the withheld ducklings than in the fed ducklings at 2 and 6 d posthatch. In conclusion, delaying access of ducklings with no experimental pathogen infection to feed and water has no long-term effects on early growth parameters, intestinal physiology, and immune responses.

Chemokine receptor CCR7 and CXCR5 mRNA in chickens following inflammation or vaccination.

The CCR7 and CXCR5 chemokine receptor mRNA contents of different immune organs were studied in normal, healthy birds and in birds treated with either lipopolysaccharide (LPS) as a systemic inflammatory challenge or coccidial vaccine (Coccivac B; Intervet/Schering-Plough Animal Health Corp., Millsboro, DE) as an enteric vaccination challenge. The CCR7 mRNA content of the spleen of normal, healthy birds was approximately 150-fold higher than CCR7 mRNA content of any other organs studied. The CXCR5 mRNA content of the bursa of normal, healthy birds was approximately 80-fold higher than the CXCR5 mRNA content of any other organs studied. The LPS injection decreased the splenic CCR7 mRNA content by approximately 100 times and the bursal CXCR5 mRNA content by approximately 5-fold at 24 h post-LPS injection (P < 0.01). The LPS injection increased the CXCR5 content of cecal tonsils by approximately 3-fold at 24 h post-LPS injection (P < 0.05). At 10 d postvaccination, CCR7 mRNA content was approximately 15-fold higher and CXCR5 mRNA content was approximately 12-fold higher in cecal tonsils of the vaccinated group than in the control group (P < 0.01). In conclusion, CCR7 and CXCR5 mRNA levels were dependent on the immune organs and the inflammatory status of the organs in chickens.

Lutein supplementation alters inflammatory cytokine production and antioxidant status in F-line turkeys.

Effect of dietary lutein supplementation on turkey production parameters, cytokine production, and oxidative status during an acute phase response following lipopolysaccharide (LPS) injection was studied. One-day-old chicks were fed a basal diet supplemented with 3 levels (0, 25, or 50 mg/kg of feed) of lutein. At 50 d of dietary lutein supplementation, turkeys were injected or not injected with LPS. Increasing dietary lutein increased the liver and plasma lutein content in both LPS injected and uninjected groups. In the groups fed 50 mg of lutein, LPS treatment decreased the lutein content of both the liver and the plasma at 48 h post-LPS injection. In the groups fed 0 mg of lutein, LPS treatment decreased the BW gain and feed consumption at 24 and 48 h post-LPS injection. The feed intake and BW gain of the group fed 50 mg of lutein in the LPS injected groups were comparable to those of the group with no LPS injection at both 24 and 48 h post-LPS injection. Treatment with LPS increased IL-1ß mRNA content (P = 0.01) in the group fed 0 mg of lutein. In the LPS injected groups, increasing dietary lutein to 50 mg decreased the IL-1ß mRNA amount compared with the group fed 0 mg of lutein. In the LPS injected groups, increasing dietary lutein to 50 mg increased IL-10 mRNA content compared with the group fed 0 mg of lutein. Injection of LPS increased the thiobarbituric reactive substances content of the liver in the group fed 0 mg of lutein. Increasing dietary lutein to 50 mg decreased the thiobarbituric reactive substances content of the liver in the LPS injected groups. Dietary lutein supplementation decreased oxidative damage and inflammatory responses post-LPS injection by decreasing IL-1ß production and increasing IL-10 production in turkeys.

Dietary lutein and fish oil interact to alter atherosclerotic lesions in a Japanese quail model of atherosclerosis.

Interactions between concentration of dietary lutein and fish oil in diets on atherosclerosis incidences were studied in a cholesterol-induced-atherosclerosis (CIA) model. CIA Japanese quail were fed a basal diet with three amounts of lutein (0, 25 and 50 mg/kg diet) and two amounts of fish oil (3% and 6%) in a 3 × 2 factorial in five replications. Samples were collected at 24 and 27 weeks of age. Atherosclerosis lesions in the dorsal aorta were measured by histochemistry sectioning. At 27 weeks of age, increasing dietary fish oil content to 6% decreased (p < 0.01) the atherosclerotic lesions only in the 0 mg lutein supplemented groups. At 27 weeks of age, increasing dietary fish oil content to 6% increased the atherosclerotic lesion score when lutein was supplemented at either 25 or 50 mg/kg feed. Aorta and liver lutein content increased (p < 0.01) with increasing dietary lutein content at 27 weeks of age. Increasing dietary fish oil content to 6% increased (p < 0.01) the aorta fat content by twofold and decreased (p < 0.01) the liver fat by 26% at 27 weeks of age. Increasing the dietary fish oil content to 6% increased (p = 0.01) the total PUFA and decreased (p = 0.03) the total mono unsaturated fatty acids content of the aorta at 27 weeks of age. At 27 weeks of age, increasing dietary fish oil content to 6% decreased the amount of TBARS (p = 0.01) and IL-1 mRNA (p < 0.01) only in the 0 mg lutein supplemented groups. Increasing dietary fish oil content to 6% increased the amount of TBARS and IL-1 mRNA of the aorta when lutein was supplemented at either 25 or 50 mg/kg diet. Dietary lutein supplementation decreased atherosclerosis lesions only at low levels of dietary polyunsaturated fatty acids.

Chicken chemokine receptors in T cells isolated from lymphoid organs and in splenocytes cultured with concanavalin A.

Chemokine receptors guide immune cells to specific organs during health and disease. The mRNA content of the chemokine receptors CCR2, CCR4, CCR5, CCR6, CCR7, CCR8, CCR9, CXCR4, CXCR5, and CX3CR1 in CD4(+) cells (T-helper cells) isolated from blood, bursa, cecal tonsil, spleen, and thymus and in CD8(+) cells (T-cytotoxic cells) isolated from blood, cecal tonsil, spleen, and thymus were investigated. The CD4(+) cells isolated from thymus had the highest amount of CCR7 and CCR8 mRNA. The CD4(+) cells isolated from bursa, cecal tonsil, and thymus had the highest amount of CCR5 mRNA. The CD4(+) cells isolated from cecal tonsils had the highest amount of CCR9 mRNA. The CD4(+) cells isolated from bursa and thymus had the highest amount of CXCR5 mRNA. The CD8(+) cells isolated from cecal tonsil had the highest mRNA amount of all receptors studied except CCR9 and CX3CR1. The CD4(+) cells treated with concanavalin A had increased CCR2, CCR4, CCR7, CCR8, and CXCR5 mRNA amounts at 24 h of stimulation. The CD8(+) cells treated with concanavalin A had increased CCR4 mRNA at 72 h, increased CCR6 mRNA at 24 h, and decreased CCR8 and CXCR4 mRNA at 24 h of stimulation.

Dietary lutein and fat interact to modify macrophage properties in chicks hatched from carotenoid deplete or replete eggs.

Two experiments were conducted to study the interaction between dietary lutein and fat levels in broiler chicks hatched from lutein depleted (Experiment I) and repleted (Experiment II) eggs. In both experiments, a 2 x 3 factorial arrangement of treatments resulted in six dietary treatments (fat at 3% and 6% and lutein at 0, 25 and 50 mg/kg feed) that were fed for 23 days to 18 birds per treatment (in three replications). In Experiment I, the anti-dinitrophenyl-keyhole-lympet-hemocyanin (anti-DNP-KLM) serum antibody response at day 22 and macrophage phagocytotic index at day 17 did not differ among treatment groups (p > 0.05). The concavalin A and phytohaemagglutinin-P lymphocyte proliferation index at day 19 was greater in birds fed 50 mg of lutein and 3% fat than in birds fed all other diets (p < 0.05). Independent of the level of dietary fat, dietary lutein increased macrophage (day 23) nitrite production measured 46 h after in vitro stimulation with LPS (p < 0.05). Among the birds fed lutein at 25 or 50 mg/kg feed, birds fed 3% fat had higher LPS-induced nitrite production compared to the birds fed 6% fat after 46 (p = 0.014) or 70 h (p < 0.001). In Experiment II, macrophage nitrite production was measured at 54 h after LPS stimulation on days 11, 15, 19 and 23. An interaction between dietary lutein and fat levels on nitrite production was observed on day 19 (p = 0.012), where macrophages from birds fed 0 mg lutein and 3% fat had the highest nitrite production (p = 0.012). Macrophages from birds fed lutein at 25 and 50 mg/kg diet and 3% fat had higher (p = 0.012) nitrite production than birds fed 6% fat. Thus, in birds hatched from lutein deplete and replete eggs, modulation of macrophage nitrite production by lutein is dependent on the level of dietary fat.

Nutritive value of full-fat sunflower seeds in broiler diets.

A study was conducted to evaluate the nutritive value and inclusion level of sunflower seed (SFS) in broiler diets. SFS contained 38.7% ether extract (EE), 16.9% CP, 14.9% crude fiber (CF), 3.5% ash, 0.57% lysine, and 0.46% methionine (89.2% DM basis). The AME (kcal/ kg) content of SFS in roosters was 5,225 and in broilers at 4, 18, and 35 d of age was 3,493, 5,132, and 5,162, respectively. The CP, EE, and CF digestibilities were 80.4, 71.2, and 11.4%, respectively. In an isocaloric and isonitrogenous diet containing SFS at 0, 5, 10, 15, and 20%, SFS up to 20% did not affect weight gain and feed consumption, but the feed conversion ratio was improved (P < 0.05) when broilers were fed 15 or 20% SFS in the starter and finisher diets. CF digestibility of starter diet was significantly lower when 15 or 20% SFS was included. CF digestibility of the finisher diet and digestibility of other nutrients in starter and finisher diets were comparable in all treatment groups. Liver and muscle lipid content, plasma total and high-density lipoprotein cholesterol content, muscle cholesterol content, dressing percentage, liver weight, and giblet weight (as % live weight) were comparable among all treatment groups. Abdominal fat was increased in birds fed 20% SFS. Percentage skin was decreased in broilers fed > or = 10% SFS.

Muscle fatty acid composition and thiobarbituric acid-reactive substances of broilers fed different cultivars of sorghum.

Two experiments were conducted to evaluate the production parameters, muscle fatty acid composition, and lipid stability of birds fed four different cultivars of sorghum. Experiment 1 used ruby red and valpo red, and Experiment 2 used kinsman and mason cultivars of sorghum. The basal diet was a corn, soy, and flax meal, and sorghum cultivars were substituted at 10 or 5% in Experiments 1 and 2. No significant difference was observed for BW, feed consumption or feed efficiency (P > 0.05). The total lipids were reduced (P < 0.05) in the leg muscle of broilers fed the mason cultivar. The TBA-reactive substances (TBARS) were lower (P < 0.05) in the leg muscle of birds fed the mason cultivar when compared to kinsman or control diet. The content of n-3 polyunsaturated fatty acids were higher (P < 0.05) in the breast and leg muscles of birds fed ruby red, kinsman, and mason cultivars of sorghum.