ABSTRACT: Changes in intestinal immunity and gut microbiota associated necrotic enteritis induced by Clostridium perfringens infection and exposure to heat stress in chicken.
Project description:Purpose: Analyze gene expression of necrotic enteritis C. perfringens in intestinal chicken loops comparing with in vitro conditions
Project description:Purpose: Analyze gene expression during C. perfringens colonization in the chicken Transcriptomic profile of mRNA from C. perfrinegns from in vivo and in vitro conditions were determined in biological duplicates by RNA-Seq using Illumina HiSeq 2500 Comparison of gene expression through RNA sequencing of necrotic enteritis C. perfrinegns type A of in vivo (chicken loops) and in vitro (lab culture)
Project description:Necrotic enteritis (NE) in broiler chickens, caused by the overgrowth of toxin-producing strains of Clostridium (C.) perfringens, results in the development of necrotic lesions, compromised intestinal health, and significant economic losses in poultry production. This study aims to analyze the blood proteome of broiler chickens affected by NE, providing insights into the host's response to the infection. Using MS/MS-based proteomics, blood plasma samples from broilers with necrotic lesions of different severity were analyzed and compared to healthy controls. A total of 412 proteins were identified, with 63 showing significant differences and (for some of those) correlating with disease severity. Gene Set Enrichment Analysis (GSEA) revealed that proteins affected by NE were predominantly associated with the immune and signaling processes and extracellular matrix (ECM) structures. Notably, regulated proteins were significantly involved in bioprocesses related to complement activation, acute phase reaction, proteolysis and humoral immune response. The findings suggest that the changes in plasma proteins in response to NE are driven by the host's intensified efforts to counteract the infection, demonstrating a.o. a notable reduction in peptides from ECM-related proteins in the blood of NE-affected birds. Overall, proteomics results underscored the attempts of the host to manage tissue damage and inflammation, indicating a coordinated effort to mitigate the pathogenic impact of C. perfringens. This study provides a deeper understanding of the host-pathogen interactions and potential targets for therapeutic intervention.
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.
Project description:Gene expression profiling of clostridium perfringens infection in broilers on medicated and non-medicated diets using chicken 44k agilent microarray. To elucidate molecular and ceelular mechanisms of bacitracin effect on CP infection in chickens by microarray technology.
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:Infectious enteritis is often accompanied with immuno-disorder of intestinal immune cells caused by microbials infection. Trained immunity is classically characterized by long-term functional reprogramming of innate immune cells to combat infectious diseases. However, whether the induction of trained immunity plays a role in protecting infectious enteritis remains largely unknown. Here, through establishing an in vivo β-glucan training and E. piscicida infection model in zebrafish, we observe that induction of trained immunity could alleviate bacterial infection-caused enteritis. Moreover, we identify intestinal complement C3 as a crucial target of trained immunity and could be amplified in response to bacterial infection. Furthermore, we reveal that trained immunity could reverse the reduction of intestinal Th17 cells in C3-dependent manner to alleviate infectious enteritis. Taken together, our results uncover the role of complement C3-mediated trained immunity in maintaining Th17 cells and intestine homeostasis, and provide a theoretical strategy for immunotherapies of infectious enteritis.
2025-05-01 | GSE285528 | GEO
Project description:Whole genome sequencing of Clostridium perfringens from healthy and necrotic enteritis affected broiler chicken farms
Project description:Comparative transcriptome analysis by RNAseq of Necrotic Enteritis Clostridium perfringens in ligated intestinal chicken loops and in vitro conditions.
Project description:Long noncoding RNAs (lncRNAs) are non-protein coding RNAs with a length of more than 200 bp, which play a vital regulatory role in intestinal immunity. Exposure to high temperature leads to death in many fish species, which is implicated in fish enteritis. Our study showed that fish enteritis was induced by acute heat stress. To date, which lncRNAs are participated in this process is still unclear. In the present study, based on the intestinal sequencing data of largemouth bass Micropterus salmoides, a total of 347,351,492 clean reads were generated from six cDNA libraries. Among them, 3399 novel lncRNA transcripts from 2488 lncRNA genes were identified. As expected, these lncRNAs exhibited shorter transcript lengths than protein-coding genes similar to those lncRNAs reported from other fish species. In total, 216 novel lncRNAs were differentially expressed (DE) in largemouth bass intestine (absolute log2 fold change > 2 and p-value < 0.05) and that 210 neighboring genes were cis-regulated by these DE-lncRNAs. An analysis of GO/KEGG enrichment showed that most of these cis-genes seemed to be significantly enriched in immune regulation (p < 0.05) and lncRNA MSTRG.8573 had an important role in regulating jak-stat signaling pathway during this process. Through this study, we showed a catalog of novel DE-lncRNAs involved in the occurrence of enteritis in largemouth bass under acute heat stress, which could provide some useful references by regulating lncRNAs to solve the heat stress-induced fish enteritis for further studies.