Project description:Recent advances in high throughput sequencing methodologies allow the opportunity to probe in depth the transcriptomes of organisms including E. tenella. In this project, we are using Illumina sequencing technology to analyze the transcriptome (RNA-Seq) of experimentally accessible stages (e.g. sporozoites, merozoites, unsporulated oocysts, sporulated oocysts) of E. tenella Houghton. The aim is to make transcriptional landscape maps of different life cycle stages of E. tenella at single base pairs resolution and utilise this information to identify the genes and define the precise gene boundaries. Gene finding has been a rather difficult task in Eimeria. Obtaining the precise transcript boundaries by Illumina RNA-Seq protocol is expected to expedite gene finding in Eimeria tenella. This data is part of a pre-publication release. For information on the proper use of pre-publication data shared by the Wellcome Trust Sanger Institute (including details of any publication moratoria), please see http://www.sanger.ac.uk/datasharing/
Project description:WFBs and the oocyst wall of Eimeria necatrix were subjected to comparative proteomic analysis using tandem mass tag (TMT) in conjunction with liquid chromatography tandem-mass spectrometry (LC-MS/MS) techniques
Project description:Whole sporozoite proteins of Eimeria acervulina were prepared and analyzed by 2-dimensional gel electrophoresis (2-DE) followed by Western blotting using immune sera specific to E. tenella, E. acervulina, or E. necatrix.
Project description:Intraspecific phenotypic variation markedly influences the damage that parasites inflict on their hosts. Such is the case for strains of Eimeria maxima, a costly enteric parasite of poultry, where strain APU-1 exhibits greater pathogenicity than APU-2. Here, we examined how these strains differ as oocysts mature to the fully-sporulated stage. We performed mi-croscopy and RNA-Sequencing on oocysts at regular intervals (6-12 hours) during sporulation. Although each strain underwent parallel development, APU-1 initially approached maturation more slowly. Each strain achieved full sporu-lation and similar transcription profiles by hour 36, after which strains appeared to diverge. These differences may in-fluence subsequent virulence. Candidate biomarkers of oocyst viability include 58 genes contributing at least 1,000 Transcripts Per Million throughout sporulation. Many genes resemble constitutively expressed genes also important to Eimeria acervulina. Mature and immature oocysts differentially express certain genes. Expression of some such bi-omarkers appears strain-specific. These data illuminate processes that may generally underlie sporulation in Eimeria and related genera, such as Cyclospora, and identify biological processes which differentiate among them. Drivers of devel-opment and senescence may provide tools to assess the viability of oocysts, which would greatly benefit the poultry industry and food safety applications.
Project description:In order to elucidate the infection mechanisms of Eimeria tenella and the chicken immune response, chicken macrophage cell cultures of cell line HD-11 were infected with Eimeria tenella strain Hougton sporozoites. Samples were taken at 0, 2, 4, 12, 24, 48 and 72 hours post-infection and, purified and mRNA sequenced. A dual-RNA seq analysis was carried out, comparing the expression of infected chicken macrophages with uninfected ones at the same time points post-infection and comparing E. tenella samples during the infection with a sample of pure sporozoites. The results show a variety of response signals in the chicken, both previously known and unknown, as well as a clear role for different sets of SAG and MIC proteins for sporozoites and merozoites of E. tenella
Project description:In order to elucidate the infection mechanisms of Eimeria tenella and the chicken immune response, chickens were infected with Eimeria tenella strain Hougton sporozoites. Samples were taken at 0, 1, 2, 3, 4 and 10 days post-infection and mRNA sequenced. A dual-RNA seq analysis was carried out, comparing the expression of infected chickens during each sampling time point with uninfected chickens and comparing E. tenella samples during the infection with a sample of pure sporozoites. The results show a variety of response signals in the chicken, both previously known and unknown, as well as a clear role for a variety of infection-related genes in E. tenella