A method for noninvasive beat-by-beat visualization of His bundle signals.

BACKGROUND: Invasive recording of His bundle signals (HBS) in electrophysiological study (EPS) is important in determining HV interval, the time taken to activate the ventricles from the His bundle. Noninvasive surface measurements of HBS are attempted by averaging typically 100-200 cardiac cycles of ECG time series in body surface potential mapping (BSPM) and in magnetocardiography (MCG) which records weak cardiac magnetic fields by highly sensitive detectors. However, noninvasive beat-by-beat extraction of HBS is challenged by ramp-like atrial signals and noise in PR segment of the cardiac cycle. METHODS: By making use of a signal-averaged trace showing prominent HBS as a guide trace, we developed a method combining interval-dependent wavelet thresholding (IDWT) and signal space projection (SSP) technique to eliminate artifacts from single beats. The method was applied on MCG recorded on 21 subjects with known HV intervals based on EPS and noninvasive signal-averaging, including five subjects with BSPM recorded subsequently. The method was also applied on stress-MCG of a subject featuring autonomic dynamics. RESULTS: HBS could be extracted from 19 out of 21 subjects by signal-averaging whose timing differed from EPS between -8 and 11 ms as tested by 2 observers. HBS in single beats were seen as aligned patterns in inter-beat contours and were appreciable in stress-MCG and conspicuous than BSPM. The performance of the method was evaluated on simulated and measured MCG to be adequate if the signal-to-noise ratio was at least 20 dB. CONCLUSIONS: These results suggest the use of this method for noninvasive assessments on HBS.

Distally Based Pedicled Fibula Flap for Reconstruction of Infected Charcot's Midtarsal Collapse-Diabetic Rocker Bottom Foot.

Introduction and Methods Diabetic rocker bottom foot with secondary infection exacts the expertise of a reconstructive surgeon to salvage the foot. The author selected 28 diabetic patients with secondarily infected Charcot's degenerated rocker bottom feet and reconstructed their feet using distally based pedicled fibula flap. Reconstruction was done in a staged manner. Stage 1 surgery involved external fixation following debridement. In stage 2, struts were activated for distraction and arthroereisis. In stage 3, the distally based pedicled fibula was used for reconstruction and beaming of the arches. Results In this retrospective study, the author analyzed the outcome of all 28 patients using the Musculoskeletal Tumor Society Rating (MSTSR) score. The average MSTSR score was 27.536 in an average follow-up of 30.5 months. The limb salvage rate with the author's procedure was 96.4% ( p = 0.045). Conclusion Author's protocol for the staged reconstruction and salvage of the infected diabetic rocker bottom foot, using the pedicled fibula flap, will be a new addendum in the reconstructive armamentarium of the orthoplastic approach.

Potential antioxidant and anti-inflammatory action of Hypericum hookerianum extracts in a liposome system evaluated with zebrafish embryos.

The purpose of this study was to assess the antioxidant and anti-inflammatory potential of liposomal formulations enriched with Hypericum hookerianum (Hyp) aqueous extracts. Cotyledon segments derived from protocorms of H. hookerianum were cultured on Murashige and Skoog (MS) media supplemented with Kinetin (KN, 1 mgl-1) and Naphthalene acetic acid (NAA, 0.1 mgl-1) to induce hypericin-rich red shoots (HypR, 0.87 mg/G DW). Highly stable liposomes (-29.4 mV) were successfully developed which encapsulated 63 ± 0.8% Hyp extracts, respectively. MTT assay subsequently confirmed the biocompatibility of liposome compositions using fibroblast cell lines. This work also evaluated acute toxicity of L-HypR and L-HypG formulations using Danio rerio (Zebrafish) embryos for 96 hpf. The expression of antioxidant and anti-inflammatory genes were found to be upregulated for L-HypR than L-HypG (green shoots without hypericin) formulations. These properties of L-HypR may be extremely useful for incorporating lipophilic substances into the food or pharmaceutical industry.

Biofilm inhibitory efficiency of phytol in combination with cefotaxime against nosocomial pathogen Acinetobacter baumannii.

AIMS: This study aimed to evaluate the antibiofilm potential of phytol and cefotaxime combinations (PCCs) against Acinetobacter baumannii and to elucidate the molecular mechanism of their antibiofilm potential through the transcriptomic approach. METHODS AND RESULTS: Phytol and cefotaxime combination(s) (PCC(s) [160 µg ml-1  + 8 µg ml-1 for microbial type culture collection (MTCC) strain and 160 µg ml-1  + 0.5 µg ml-1 for clinical isolate] effectively inhibited the A. baumannii biofilm formation. Additionally, light, confocal laser scanning and scanning electron microscopic analyses validated the antibiofilm potential of PCCs. Furthermore, PCCs treated A. baumannii cells showed a decreased level of hydrophobicity index compared to their respective controls. Fourier-transform infrared (FT-IR) spectra of exopolysaccharide matrix extracted from PCCs-treated A. baumannii cells showed a visible decrease in absorbance of polysaccharides, nucleic acids and protein regions compared to the spectra of untreated controls. In the blood sensitivity assay, the PCCs-treated A. baumannii plates showed reduced a number of bacterial colonies compared to their control plates. Reduced level of catalase production was also observed in the PCCs treatment compared to their controls. Transcriptomic analysis revealed the downregulation of bfmR, bap, csuA/B, ompA, pgaA, pgaC and katE biofilm virulence genes in both the A. baumannii strains on treatment with PCCs. CONCLUSION: The obtained results of this study indicate that PCCs have potent antibiofilm activity and downregulate the biofilm-related virulence genes expression in A. baumannii. SIGNIFICANCE AND IMPACT OF THE STUDY: To the best of our knowledge, this is the pioneering study, which shows the antibiofilm effect of PCCs against A. baumannii along with their molecular mechanism. The antibiofilm effect of PCCs could be a successful strategy for eradicating infections related to A. baumannii biofilms in nosocomial settings.

Application of cellular automata approach for cloud simulation and rendering.

Current techniques for creating clouds in games and other real time applications produce static, homogenous clouds. These clouds, while viable for real time applications, do not exhibit an organic feel that clouds in nature exhibit. These clouds, when viewed over a time period, were able to deform their initial shape and move in a more organic and dynamic way. With cloud shape technology we should be able in the future to extend to create even more cloud shapes in real time with more forces. Clouds are an essential part of any computer model of a landscape or an animation of an outdoor scene. A realistic animation of clouds is also important for creating scenes for flight simulators, movies, games, and other. Our goal was to create a realistic animation of clouds.

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.

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.

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.

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.

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.

Clinical relevance of double-arm blood pressure measurement and prevalence of clinically important inter-arm blood pressure differences in Indian Primary Care.

Hypertension guidelines recommend measuring blood pressure (BP) in both arms at least once. However, this is seldom done due to uncertainties regarding measurement procedure and the implications of finding a clinically important inter-arm BP difference (IAD). This study aimed to provide insight into the prevalence of clinically important IADs in a large Indian primary care cohort. A number of 134678 (37% female) unselected Indian primary care participants, mean age 45.2 (SD 11.9) years, had BP measured in both arms using a standardized, triplicate, automated simultaneous measurement method (Microlife WatchBP Office Afib). On average, there were clinically minor differences in right and left arm BP values: systolic BP 134.4 vs. 134.2 mmHg (p<0.01) and diastolic BP 82.7 vs. 82.6 mmHg (p<0.01), respectively. Prevalence of significant mean systolic IAD between 10 and 15 mmHg was 7813 (5.8%). Systolic IAD ≥ 15 mmHg 2980 (2.2%) and diastolic IAD ≥ 10 mmHg 7151 (5.3%). In total, there were 7595 (5.6%) and 8548 (6.3%) participants with BP above the 140/90 mmHg threshold in only the left or right arm, respectively. Prevalence of participants with elevated BP on one arm only was highest in patients with a systolic IAD ≥ 15 mmHg; 19.1% and 13.7%, for left and right arm, respectively. This study shows that a substantial prevalence of IAD exist in Indian primary care patients. BP is above the diagnostic threshold for hypertension in one arm only for 6% of participants. These findings emphasise the importance of undertaking bilateral BP measurement in routine clinical practice. This article is protected by copyright. All rights reserved.

Clinical relevance of double-arm blood pressure measurement and prevalence of clinically important inter-arm blood pressure differences in Indian primary care.

Hypertension guidelines recommend measuring blood pressure (BP) in both arms at least once. However, this is seldom done due to uncertainties regarding measurement procedure and the implications of finding a clinically important inter-arm BP difference (IAD). This study aimed to provide insight into the prevalence of clinically important IADs in a large Indian primary care cohort. A number of 134 678 (37% female) unselected Indian primary care participants, mean age 45.2 (SD 11.9) years, had BP measured in both arms using a standardized, triplicate, automated simultaneous measurement method (Microlife WatchBP Office Afib). On average, there were clinically minor differences in right and left arm BP values: systolic BP 134.4 vs 134.2 mmHg (p < .01) and diastolic BP 82.7 vs 82.6 mmHg (p < .01), respectively. Prevalence of significant mean systolic IAD between 10 and 15 mmHg was 7,813 (5.8%). Systolic IAD ≥ 15 mmHg 2,980 (2.2%) and diastolic IAD ≥ 10 mmHg 7,151 (5.3%). In total, there were 7,595 (5.6%) and 8,548 (6.3%) participants with BP above the 140/90 mmHg threshold in only the left or right arm, respectively. Prevalence of participants with elevated BP on one arm only was highest in patients with a systolic IAD ≥ 15 mmHg; 19.1% and 13.7%, for left and right arm, respectively. This study shows that a substantial prevalence of IAD exists in Indian primary care patients. BP is above the diagnostic threshold for hypertension in one arm only for 6% of participants. These findings emphasize the importance of undertaking bilateral BP measurement in routine clinical practice.

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.

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.

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.

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.

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.