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Expression profiles of genes in Toll-like receptor-mediated signaling of broilers infected with Clostridium perfringens.

ABSTRACT: Toll-like receptors (TLRs) participate in detecting microbial pattern molecules for activation of the host immune response. We investigated possible roles of TLRs in the chicken response to Clostridium perfringens infection by examining the expression of TLR genes and other genes involved in TLR-mediated signaling within the spleens and ilea of C. perfringens-challenged broilers. Upregulation of a tumor necrosis factor alpha-inducing factor homolog in challenged chickens compared to naïve chickens was observed, regardless of the incidence of necrotic enteritis. In addition, the members of the TLR2 subfamily were found to be most strongly involved in the host response to C. perfringens challenge, although the expression of TLR4 and TLR7 was also upregulated in spleen tissues. While the combination of TLR1.2, TLR2.1, and TLR15 appeared to play a major role in the splenic response, the expression of TLR2.2 and TLR1.1 was positively correlated to the expression of adaptor molecules MyD88, TRAF6, TRIF, and receptor interacting protein 1 in the ileal tissues, demonstrating a dynamic spatial and temporal innate host response to C. perfringens.

SUBMITTER: Lu Y

PROVIDER: S-EPMC2772368 | biostudies-literature | 2009 Nov

REPOSITORIES: biostudies-literature

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Expression profiles of genes in Toll-like receptor-mediated signaling of broilers infected with Clostridium perfringens.

Lu Yang Y Sarson Aimie J AJ Gong Joshua J Zhou Huaijun H Zhu Weiyun W Kang Zhumei Z Yu Hai H Sharif Shayan S Han Yanming Y

Clinical and vaccine immunology : CVI 20090923 11

Toll-like receptors (TLRs) participate in detecting microbial pattern molecules for activation of the host immune response. We investigated possible roles of TLRs in the chicken response to Clostridium perfringens infection by examining the expression of TLR genes and other genes involved in TLR-mediated signaling within the spleens and ilea of C. perfringens-challenged broilers. Upregulation of a tumor necrosis factor alpha-inducing factor homolog in challenged chickens compared to naïve chicke ...[more]

PMID: 19776194

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Project description:Necrotic enteritis is a disease caused by Clostridium perfringens, which threatens poultry production in the absence of dietary antibiotics. A total number of 144 Ross broilers were reared in 12 pens with each hosting 12 birds. Each 6 pens of birds were fed medicated (bacitracin at 55 ppm) or non-medicated starter diets (Nutreco Canada Agresearch) immediately after the chicks were placed. At day 18, birds were challenged with C. perfringens (107 cfu per ml mixed with feed). Spleens were collected from 12 birds of each group at day 18 (before infection), 19, 20, and 22. A low-density chicken immune microarray was used to study gene expression profiling of host response to C. perfringens infection. Six biological replicates (2 birds per biological replicate) for each treatment group were labeled with either Cy5 or Cy3 with dye swap. A total of 24 arrays were used for this study. Gene signal intensity was globally normalized by LOWESS and expressed as log2 ratios. A mixed model including treatment, time, array, subgrid (random effect), dye, and all interactions among treatment and time was used to identify differentially expressed genes between post-infection vs. pre-infection, among post-infections, and between medication treatments, at the 5% significance level. The results indicated subtle medication effects on gene expression of these immune-related genes compared to bacterial infection effect. Our findings strongly suggest that both cell-mediated and antibody-mediated immune responses via MHC class I and II systems were actively involved in the host defense against C. perfringens infection in broilers. The unique cytokine signaling pathway and apoptosis cascade found in the study provide a new insight of molecular regulation of host immune response. Collectively, the findings of the present study will shed light on the molecular mechanisms underlying C. perfringens infection in broilers. There were two groups: medicated and non-medicated. Spleen were collected to isolate total RNA for gene expression profiling.For the microarray study, two birds from each pen were pooled within each group. To account for any bias inherent to the fluorescent dyes, three of the six medicated replicates were labeled with Cy3 and the other three were labeled with Cy5 at each time point. The same design was applied for the non-medicated group. There were six hybridizations between medicated and non-medicated replicates at each time point. Twenty-three out of 24 arrays were used (data from one array were discarded due to poor quality).

Project description:Simple Summary Coccidia and C. perfringens are the main accomplices in broiler necrotizing enteritis (NE), and NE causes oxidative stress and intestinal damage in broilers, which will result in huge economic losses to the poultry industry. Vitamin A (VA) is an essential micronutrient of the diet, and the beneficial effects of VA on vision, growth, and development have been intensively investigated. At present, the effects of VA on intestinal barrier and antioxidant functions in broilers have not been systematically reported. The present study was conducted to investigate the effects of dietary supplementation with vitamin A on the antioxidant and intestinal barrier function of broilers co-infected with coccidia and C. perfringens (CCP). Our outcomes showed that dietary VA might help a little with intestinal barrier function; nonetheless, it failed to alleviate the negative effects of CCP on the antioxidant function in broilers. Our study has guiding significance for the dose of VA in the diet of broilers, and it might arouse readers’ strong interest. Abstract Necrotic enteritis (NE) impairs poultry production and causes great economic loss. The nutritional regulation of diets has the potential to alleviate NE. The present study was conducted to investigate the effects of dietary supplementation with vitamin A (VA) on the antioxidant and intestinal barrier function of broilers co-infected with coccidia and C. perfringens (CCP). In a 2 × 2 factorial arrangement, 336 one-day-old Ross 308 broilers were divided into four treatments with two levels of VA (0 or 12,000 IU/kg) and challenged with or without CCP. The animal trial lasted for 42 days. The results showed that dietary supplemental VA improved body weight gain (BWG) and the feed intake (FI), and the FI was negatively affected by CCP. Additionally, the levels of catalase (CAT) in the serum, total superoxide dismutase (T-SOD), and CAT in the jejunum and glutathione peroxidase (GSH-Px) in the liver decreased with the CCP challenge (p < 0.05). The mRNA levels of SOD, CAT, GSH-Px1, and GSH-Px3 in the liver and jejunum were upregulated by the CCP challenge (p < 0.05). In addition, the level of serum diamine oxidase (DAO), and the mRNA level of ZO-1 were also upregulated with the CCP challenge. Dietary supplementation with VA contributed to the intestinal villi height and the mRNA level of Mucin-2 in the jejunum (p < 0.05). Additionally, dietary VA had the ability to alleviate the upregulation of SOD in the liver and SOD, CAT, GSH-Px1, GSH-Px3, ZO-1, and claudin-1 in the jejunum with the CCP challenge (p < 0.05). However, the mRNA level of GSH-Px3 and the levels of SOD in the liver and jejunum were downregulated with the VA supplementation in the diet. In conclusion, dietary VA improved the growth performance and the intestinal barrier function; nonetheless, it failed to alleviate the negative effects of CCP on the antioxidant function in broilers.

Project description:BackgroundBacteriophages are viral predators of bacteria and are common in nature. Their host-specific infections against specific bacteria make them an attractive natural agent to control bacterial pathogens. Interest in the potential of bacteriophages as antibacterial agents in the production animal industries has increased.MethodsA total of 18 bacteriophages were isolated from Australian commercial poultry environments, from which three highly active phages were chosen for enrichment. Sequencing libraries were prepared using a Nextera XT kit (Illumina) and sequenced on an Illumina MiSeq instrument using 2 × 300 bp paired-end chemistry. The sequence data were then assembled and aligned with a2 bacteriophage as the reference. An animal trial was performed by oral gavaging Clostridium perfringens netB containing strain EHE-NE18 to the Ross 308 broiler chickens prior inoculation with Eimeria species. The chickens were raised following the management guide for Ross 308 from d 0 to d 21 and fed with starter and grower diets met the specific breed nutrient requirements. Body weight gain and feed intake were measured on d 9 and d 21 and FCR adjusted with mortality was calculated.ResultsThe isolated bacteriophages only had only 96.7% similarity to the most closely related, previously characterized, Clostridium bacteriophage indicated that they might represent a novel strain of bacteriophage. A "cocktail" containing the three bacteriophages was capable of lysing four known disease-inducing C. perfringens strains in vitro. Oral administration of the bacteriophages cocktail to broilers challenged with necrotic enteritis markedly alleviated intestinal necrotic lesions in the duodenum and jejunum on day 16 post-hatch. The phage treatment significantly reduced the lesion scores of birds challenged with NE (P < 0.01), and the lesion scores between birds treated with the bacteriophages and the unchallenged birds were not statistically different (P > 0.05). However, no effect on the growth performance was observed during the recorded period of days 9-21.ConclusionThese findings suggest that bacteriophage treatment is a promising approach to protect intestinal health from C. perfringens induced necrotic enteritis. Further research will be required on the dosing, route of administration, and large scale validation studies to further advance this approach to pathogen control.

Dataset Information

Expression profiles of genes in Toll-like receptor-mediated signaling of broilers infected with Clostridium perfringens.

Publications

Expression profiles of genes in Toll-like receptor-mediated signaling of broilers infected with Clostridium perfringens.

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