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.

Short-term exposure to fumonisins and deoxynivalenol, on broiler growth performance and cecal Salmonella load during experimental Salmonella Enteritidis infection.

Fumonisins (FUM) and deoxynivalenol (DON) are two common mycotoxins in poultry feed. Salmonella enterica ser. Enteritidis (S. Enteritidis) is a primary foodborne bacterium in broilers. This trial was conducted to evaluate the effects of naturally occurring FUM and DON and their combination at subclinical doses on broiler performance during a S. Enteritidis challenge. The experiment consisted of five treatments: NCC, no-challenge no-mycotoxin treatment; CC, Salmonella challenge + no-mycotoxin treatment; DON, DON 0.6 mg/kg + Salmonella challenge; FUM, FUM 14 mg/kg + Salmonella challenge; DON + FUM + T-2 + neosolaniol, DON 0.6 mg/kg + FUM 14 mg/kg + T-2 toxin 0.6 mg/kg + 0.8 mg/kg neosolaniol + Salmonella challenge. On d 4, birds were challenged with either 0 or 1 × 109 CFU/mL S. Enteritidis orally. There were no significant effects on growth performance among treatments at 0, 3, 7, and 14 d of post-inoculation (dpi). On 14 dpi, the combined DON + FUM + T-2 + neosolaniol significantly increased the Salmonella load by 1.5 logs compared to the control groups (P < 0.05). FUM significantly increased the cecal tonsil IL-10 gene expression by 1.2-fold at 7 dpi (P < 0.05) and downregulated TNF-α by 1.8-fold on 14 dpi compared to the control, nonchallenge groups (P < 0.05). On 7 dpi, the combined DON + FUM + T-2 + neosolaniol reduced occludin by 4.4-fold (P < 0.05) when compared to the control groups. Similarly, combined DON + FUM+ T-2 + neosolaniol decreased zona-occluden transcription by 2.3 and 7.6-fold on 3 and 14 dpi, respectively (P < 0.05). Furthermore, combined DON + FUM + T-2 + neosolaniol decreased Claudin-1 by 2.2-fold and Claudin-4 by 5.1-fold on 14 dpi when compared to the control groups (P < 0.05). In conclusion, short-term exposure to a subclinical dose of combined DON + FUM + T-2 + neosolaniol had an impact on broiler intestinal tight junction proteins and cecal Salmonella abundance under experimental Salmonella challenge.

Subclinical Doses of Combined Fumonisins and Deoxynivalenol Predispose Clostridium perfringens-Inoculated Broilers to Necrotic Enteritis.

Fumonisins (FB) and deoxynivalenol (DON) are mycotoxins which may predispose broiler chickens to necrotic enteritis (NE). The objective of this study was to identify the effects of subclinical doses of combined FB and DON on NE. A total of 480 day-old male broiler chicks were divided into four treatment groups; 1) control group (basal diet + Clostridium perfringens); 2) necrotic enteritis group (basal diet + Eimeria maxima + C. perfringens); 3) FB + DON group (basal diet + 3 mg/kg FB + 4 mg/kg DON + C. perfringens); and 4) FB + DON + NE group (basal diet + 3 mg/kg FB + 4 mg/kg DON + E. maxima + C. perfringens). Birds in NE and FB + DON + NE groups received 2.5 × 103 E. maxima on day 14. All birds were inoculated with C. perfringens on days 19, 20, and 21. On day 35, birds in the NE, FB + DON, and FB + DON + NE groups had 242, 84, and 339 g lower BWG and a 19-, 2-, and 22-point increase in FCR respectively, than in the control group. Subclinical doses of FB + DON increased (p < 0.05) the NE lesion scores compared to the control group on day 21. On day 21, birds in the NE, FB + DON, and FB + DON + NE groups had increased (p < 0.05) serum FITC-D, lower (p < 0.05) jejunal tight junction protein mRNA, and increased (p < 0.05) cecal tonsil IL-1 mRNA compared to control group. On day 21, birds in the NE group had decreased (p < 0.05) villi height to crypt depth ratio compared to the control group and the presence of FB + DON in NE-induced birds further decreased the villi height to crypt depth ratio. Birds in the NE, FB + DON, and FB + DON + NE groups had increased (p < 0.05) C. perfringens, lower (p < 0.05) Lactobacillus loads in the cecal content, and a lower (p < 0.05) CD8+: CD4+ cell ratio in the cecal tonsils compared to the control group. It can be concluded that subclinical doses of combined FB and DON predispose C. perfringens-inoculated birds to NE, and the presence of FB + DON in NE-induced birds exacerbated the severity of NE.

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.

Effect of a post-hatch lipopolysaccharide challenge in Turkey poults and ducklings after a primary embryonic heat stress.

The effect of embryonic thermal manipulation on the post-hatch immune response to a lipopolysaccharide (LPS) challenge was studied in Pekin ducklings and turkey poults. Commercial duck and turkey eggs were distributed among four treatments: SS-Control (37.5 °C from embryonic day [ED] 1 to 25); SS-LPS (37.5 °C from ED1 to 25 + LPS at D0 [hatch]); HH-LPS (38 °C from ED1 to 25 + LPS at D0; SH-LPS (37.5 °C from ED1 to 10 and 38 °C from ED 11 to 25 + LPS at D0). At ED16 and ED24, the egg shell temperature of the duck and turkey eggs in the HH and SH treatments were higher (P ≤ 0.01) than the SS treatment. Ducklings and poults in the HH treatment had the lowest yolk free body weight at hatch (P ≤ 0.05). At 24, 48, and 72 h post-LPS injection, ducklings and poults in the HH-LPS treatment had significantly reduced BW compared with the SS-Con treatments (P ≤ 0.05). Ducklings and poults in the SH-LPS and HH-LPS treatments had increased plasma heat shock protein 70 (HSP70) and lower splenic HSP70 mRNA amounts than the SS-LPS treatments at 24, and 48 h post-challenge (P ≤ 0.05). At 48 and 72 h, macrophage nitric oxide (NO) production in ducklings and poults in the SH-LPS and HH-LPS treatments was lower than in the SS-LPS treatments (P ≤ 0.05). Ducklings and poults in the SH-LPS treatment had increased thymocyte proliferation compared to the SS-LPS treatment at 24, 48 and 72 h (P ≤ 0.05). At 24 h, ducklings in the SH-LPS treatment had increased splenic IL-10 and reduced IFNγ and IL-6 mRNA abundance. However, both ducklings and poults in the HH-LPS treatment had increased IFNγ, and IL-10 mRNA abundance compared to the SS-LPS treatment (P ≤ 0.05). At 48 h, SH-LPS ducklings and poults had lower splenic IL-10 mRNA abundance (P ≤ 0.05) while the HH-LPS treatment resulted in comparable splenic IL-10 mRNA compared to the SS-LPS treatment (P ≥ 0.05). Ducklings and poults in the SH-LPS treatment had increased thymic and splenic CD8+/CD4+ ratios at 24 h versus the SS-LPS treatment (P ≤ 0.05). In conclusion, embryonic thermal manipulation from ED11-25 increased extracellular HSP70 release, thymocyte proliferation and IL-10 but decreased splenic HSP70 and IFNγ mRNA amounts at 24 h post-LPS injection. This suggests that mild heat stress during the later stages of incubation could potentially prime the embryonic immune system thereby enhances the immune response as earlier than 24 h to eliminate the inflammatory response without affecting the growth performance by increase the extracellular release of HSP70 in both ducklings and poults. Continuous exposure to the small increase in temperature from ED 1-25 (HH) caused an imbalance between pro (IFNγ)- and anti-inflammatory cytokines(IL-10) which affects hatchling responses to an inflammatory challenge and increased mortality. The amount of extracellular HSP70 could potentially play an important role in modulating the immune response against inflammatory challenges.

Effect of embryonic thermal manipulation on heat shock protein 70 (HSP70) expression and subsequent immune response to post-hatch lipopolysaccharide challenge in Pekin ducklings.

During the course of multi-stage incubation, small locational differences in incubation temperature within a machine are not uncommon and so the goal of this study was to study the immune response of ducklings exposed to thermal manipulation during incubation. Commercial Pekin duck eggs (n = 200) were distributed among four treatment: SS-Control (37.5°C from embryonic day [ED] 1 to 25); SS-LPS (37.5°C from ED1 to 25 + LPS at D0 [hatch]); HH-LPS (38°C from ED1 to 25+ LPS at D0); SH-LPS (37.5°C from ED1 to 10 and 38°C from ED 11 to 25 + LPS at D0). At D0, ducklings received a lipopolysaccharide (LPS) injection. At D1 and D5, the HH-LPS treatment significantly reduced body weight (P ≤ 0.05). At D1 and D3 post-LPS injection, the SH-LPS and HH-LPS treatments significantly reduced splenic and bursal heat shock protein 70 (HSP70), mRNA abundance, and macrophage nitric oxide production compared with the SS-LPS treatment (P ≤ 0.05). At D1, the HH-LPS and SH-LPS treatments had increased splenic IL-10 mRNA and lower MHC I mRNA compared with the SS-LPS treatment (P ≤ 0.05). At D1, the HH-LPS treatment increased splenic IL-6 mRNA and bursal IFNγ mRNA transcription while the SH-LPS treatment reduced splenic IL-6 mRNA compared with the SS-LPS treatment (P ≤ 0.05). The HH-LPS treatment reduced thymocyte proliferation efficiency, while at D1, D3, and D5, the SH-LPS treatment increased thymocyte proliferation efficiency compared with the SS-LPS treatment (P ≤ 0.05). Ducklings in the HH-LPS treatment had a higher splenic CD8+/CD4+ ratio compared to the SS-LPS treatment at D3 post-LPS injection (P ≤ 0.05). In summary, the HH-LPS treatment compromised immunocompetence via decreased NO production and thymocyte proliferation efficiency, while the SH-LPS treatment increased body weight and thymocyte proliferation and reduced IL-6 mRNA abundance. This suggests that an embryonic temperature stress during the latter half of incubation may prime the immune system which may be beneficial during secondary post-hatch inflammatory challenges.

Effect of synbiotic supplementation on layer production and cecal Salmonella load during a Salmonella challenge.

This study analyzed the inhibitory effects of a synbiotic product (PoultryStar® me) on production parameters, intestinal microflora profile, and immune parameters in laying hens with and without a Salmonella challenge. The synbiotic product contained 4 probiotic bacterial strains (Lactobacillus reuteri, Enterococcus faecium, Bifidobacterium animalis, and Pediococcus acidilactici) and a prebiotic fructooligosaccharide. Layers were supplemented with the synbiotic from d of hatch to 28 wk of age. At 16 wk of age, birds were either vaccinated with Salmonella enterica Enteritidis (SE) vaccine or left unvaccinated. At 24 wk of age, a portion of the birds was challenged with 1 × 109 CFU of SE or left unchallenged, resulting in a 3 (vaccinated, challenged, or both vaccinated and challenged) X 2 (control and synbiotics) factorial arrangement of treatments. At 18 and 20 wk of age, birds fed synbiotics in both vaccinated and unvaccinated groups had increased (P < 0.05) BW more than those in the un-supplemented groups. Birds fed synbiotics had 0.7, 17.8, 21.7, 3, and 4.2% higher (P < 0.05) hen d egg production (HDEP) at 19, 20, 21, 22, and 23 wk of age, compared to the birds without supplementation, respectively. After administering the SE challenge, supplemented birds had 3, 6.7, 4.3, 12.5, and 14.4% higher (P < 0.05) HDEP at 24, 25, 26, 27, and 28 wk of age, compared to the birds not supplemented, respectively. Irrespective of the vaccination status, birds fed synbiotics and challenged with SE had a lower (P < 0.05) SE cecal load compared to the un-supplemented groups. At 22 d post Salmonella challenge, birds supplemented, vaccinated, and challenged had the highest bile IgA content. It can be concluded that supplementation of the synbiotic product could be beneficial to layer diets as a growth promoter, performance enhancer, and for protection against SE infection.

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.

Uterine Arteriovenous Malformation: Case Series and Literature Review.

BACKGROUND: Uterine AV malformation is a rare cause of torrential post-abortal hemorrhage, which can present with varying grades of severity. Diagnosis requires a high degree of suspicion and is done with ultrasound and Doppler. CASE SERIES: In our institution, during the period 2008-2013, five cases of symptomatic uterine AVMs have been reported. All of them were in the reproductive age group (22-36 years), presenting with a history of miscarriage or termination of pregnancy for which curettage was done. The presentation was with recurrent bouts of torrential bleeding, some triggered by second curettage, and not controllable with regular measures. Diagnosis was by ultrasound-gray scale, color Doppler, and spectral Doppler. The time interval between the onset of symptoms and the primary curettage was 8-89 days; four patients underwent selective arterial embolization, and one patient opted for hysterectomy. On follow-up, all the four patients are presently free of symptoms; two of them conceived within 2 years of the procedure and carried the pregnancy to term-one resulting in a live-birth and the other intrauterine death. CONCLUSION: Uterine AV malformation should be thought of as a differential diagnosis in all cases presenting with bleeding after miscarriage or curettage, since diagnosis is simple and treatment by selective arterial embolization saves morbidity of surgery and anesthesia, and more importantly reduces hospital stay and the absence from work.

Comparison of Carotid Intima Media Thickness in Children of Patients with and without Premature Coronary Artery Disease.

INTRODUCTION: Increased Carotid Intima Medial Thickness (CIMT) is associated with cardiovascular risk factors and vascular events like Coronary Artery Disease (CAD) and stroke. AIM: This study was designed to identify whether CIMT is increased in the children of parents with premature coronary artery disease and compare it with age and sex matched controls who are children of normal individuals. We also tried to compare and correlate the changes in CIMT if any, among the study group with relation to family history of diabetes and hypertension. MATERIALS AND METHODS: It was an observational prospective case control study. Twenty five cases (children of parents with premature coronary artery disease) were recruited as per the inclusion and exclusion criteria. Age and sex matched controls were recruited from the paediatric Outpatient Department (OPD). The CIMT was measured using ultrasound Doppler as per the protocol by the Mannheim intima media thickness consensus statement. RESULTS: Ten children out of 25 controls had CIMT of 0.05 cm and 5 out of 25 cases had CIMT of 0.07 cm. The association between cases and controls was not statistically significant. Even though the CIMT increases after 10 years, there is no statistically significant association between cases and controls in the different age groups. Out of the total 48% of the children among the case group had family history of hypertension whereas 28% in the control group. A 33% of children in the case group with family history of hypertension had CIMT of 0.07 cm; 46% of children in case group without family history of hypertension had CIMT of 0.05 cm. This difference was statistically significant (p=0.005). There was no significant association between family history of diabetes mellitus and CIMT in both groups. CONCLUSION: There was significant association between CIMT in children with family history of premature CAD and family history of hypertension when compared with no family history of hypertension with premature CAD. There was no significant association between CIMT of children with and without family history of premature CAD.

Fly ash based Bacillus thuringiensis var. israelensis formulation for use against Culex quinquefasciatus, the vector of filariasis in natural ecosystems.

BACKGROUND & OBJECTIVES: Fly ash is produced in huge quantities by the various thermal power stations in India. This thermal waste has been employed as a carrier material in the preparation of a biopesticidal water dispersible powder (WDP) formulation for use against mosquitoes. In the present investigation, this newly developed fly ash based WDP formulation was evaluated in natural breeding habitats of mosquito. METHODS: Fly ash based WDP formulation of Bacillus thuringiensis var. israelensis (VCRC B17) was evaluated for its efficacy and residual activity in aquatic habitats supporting breeding of Culex quinquefasciatus, the vector of lymphatic filariasis in Neyveli Township, Neyveli Lignite Corporation, India for a period of one month. RESULTS: At an application rate of 10 kg/ha, the WDP was effective for five days regardless of the habitat, and provided 80-100% reduction in larval abundance of Cx. quinquefasciatus. INTERPRETATION & CONCLUSION: The study indicates that for continued control of immature density and prevention of adult emergence, a weekly application of this formulation is necessary. This study also showed that fly ash based formulations can be used for immediate control of mosquitoes in different types of habitats and has also brought out a new avenue for the utilization of coal ash.

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.

An uncommon presentation of a rare disease - high-degree AV block with takotsubo cardiomyopathy.

Takotsubo cardiomyopathy is a type of non-ischemic cardiomyopathy in which there is sudden temporary left ventricular dysfunction. High-degree AV block and takotsubo cardiomyopathy have been reported together rarely in medical literature. Here we discuss a case of takotsubo cardiomyopathy presenting with complete heart block. A 72-year-old female presented with retrosternal chest pain. Electrocardiogram showed complete heart block without any significant ST-T changes. Echocardiogram revealed regional wall motion abnormality not consistent with coronary artery disease and was suggestive of apical ballooning. Coronary angiogram showed no significant coronary artery lesion. LV angiogram showed apical ballooning and LV systolic dysfunction. Patient underwent temporary pacemaker implantation. Since the complete heart block did not revert even after 18 days, she underwent a permanent pacemaker implantation.

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.

Evaluation of certain insecticides on nettings for their efficacy and wash resistance against mosquito species.

Five insecticides (Bifenthrin, Deltamethrin, Etofenprox, Permethrin and Lamda cyhalothrin) recommended by WHO, at their recommended dose were compared for their efficacy and wash resistance through bioassay against mosquito vectors, Culex quinquefasciatus, Aedes aegypti and Anopheles stephensi. Etofenprox treated nettings exhibited better knockdown and mortality than the other insecticides. The order of efficacy of the insecticides treated nettings was Etofenprox > or = Deltamethrin > Lambda cyhalothrin > Permethrin > Bifenthrin.

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.