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      RNA Seq analysis of the Eimeria tenella gametocyte transcriptome reveals clues about the molecular basis for sexual reproduction and oocyst biogenesis

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          Abstract

          Background

          The protozoan Eimeria tenella is a common parasite of chickens, causing avian coccidiosis, a disease of on-going concern to agricultural industries. The high prevalence of E. tenella can be attributed to the resilient oocyst stage, which is transmitted between hosts in the environment. As in related Coccidia, development of the eimerian oocyst appears to be dependent on completion of the parasite’s sexual cycle. RNA Seq transcriptome profiling offers insights into the mechanisms governing the biology of E. tenella sexual stages (gametocytes) and the potential to identify targets for blocking parasite transmission.

          Results

          Comparisons between the sequenced transcriptomes of E. tenella gametocytes and two asexual developmental stages, merozoites and sporozoites, revealed upregulated gametocyte transcription of 863 genes. Many of these genes code for proteins involved in coccidian sexual biology, such as oocyst wall biosynthesis and fertilisation, and some of these were characterised in more depth. Thus, macrogametocyte-specific expression and localisation was confirmed for two proteins destined for incorporation into the oocyst wall, as well as for a subtilisin protease and an oxidoreductase. Homologues of an oocyst wall protein and oxidoreductase were found in the related coccidian, Toxoplasma gondii, and shown to be macrogametocyte-specific. In addition, a microgametocyte gamete fusion protein, EtHAP2, was discovered.

          Conclusions

          The need for novel vaccine candidates capable of controlling coccidiosis is rising and this panel of gametocyte targets represents an invaluable resource for development of future strategies to interrupt parasite transmission, not just in Eimeria but in other Coccidia, including Toxoplasma, where transmission blocking is a relatively unexplored strategy.

          Electronic supplementary material

          The online version of this article (doi:10.1186/s12864-015-1298-6) contains supplementary material, which is available to authorized users.

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          Most cited references57

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          Discovery of gene function by expression profiling of the malaria parasite life cycle.

          The completion of the genome sequence for Plasmodium falciparum, the species responsible for most malaria human deaths, has the potential to reveal hundreds of new drug targets and proteins involved in pathogenesis. However, only approximately 35% of the genes code for proteins with an identifiable function. The absence of routine genetic tools for studying Plasmodium parasites suggests that this number is unlikely to change quickly if conventional serial methods are used to characterize encoded proteins. Here, we use a high-density oligonucleotide array to generate expression profiles of human and mosquito stages of the malaria parasite's life cycle. Genes with highly correlated levels and temporal patterns of expression were often involved in similar functions or cellular processes.
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            Count-based differential expression analysis of RNA sequencing data using R and Bioconductor

            , , (2013)
            RNA sequencing (RNA-seq) has been rapidly adopted for the profiling of transcriptomes in many areas of biology, including studies into gene regulation, development and disease. Of particular interest is the discovery of differentially expressed genes across different conditions (e.g., tissues, perturbations), while optionally adjusting for other systematic factors that affect the data collection process. There are a number of subtle yet critical aspects of these analyses, such as read counting, appropriate treatment of biological variability, quality control checks and appropriate setup of statistical modeling. Several variations have been presented in the literature, and there is a need for guidance on current best practices. This protocol presents a "state-of-the-art" computational and statistical RNA-seq differential expression analysis workflow largely based on the free open-source R language and Bioconductor software and in particular, two widely-used tools DESeq and edgeR. Hands-on time for typical small experiments (e.g., 4-10 samples) can be <1 hour, with computation time <1 day using a standard desktop PC.
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              Poultry coccidiosis: recent advancements in control measures and vaccine development.

              Coccidiosis is recognized as the major parasitic disease of poultry and is caused by the apicomplexan protozoan Eimeria. Coccidiosis seriously impairs the growth and feed utilization of infected animals resulting in loss of productivity. Conventional disease control strategies rely heavily on chemoprophylaxis and, to a certain extent, live vaccines. Combined, these factors inflict tremendous economic losses to the world poultry industry in excess of USD 3 billion annually. Increasing regulations and bans on the use of anticoccidial drugs coupled with the associated costs in developing new drugs and live vaccines increases the need for the development of novel approaches and alternative control strategies for coccidiosis. This paper aims to review the current progress in understanding the host immune response to Eimeria and discuss current and potential strategies being developed for coccidiosis control in poultry.
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                Author and article information

                Contributors
                robert.walker1@jcu.edu.au
                philippasharman@gmail.com
                kate.miller1@jcu.edu.au
                christoph.lippuner@dkf.unibe.ch
                michal.okoniewski@gmail.com
                ramon.eichenberger@uzh.ch
                chandra.ramakrishnan@uzh.ch
                nicholas.smith@jcu.edu.au
                deplazesp@access.uzh.ch
                adrian.hehl@uzh.ch
                nicholas.smith@jcu.edu.au
                Journal
                BMC Genomics
                BMC Genomics
                BMC Genomics
                BioMed Central (London )
                1471-2164
                18 February 2015
                18 February 2015
                2015
                : 16
                : 1
                : 94
                Affiliations
                [ ]Queensland Tropical Health Alliance Research Laboratory, Australian Institute of Tropical Health and Medicine, James Cook University, Cairns Campus, McGregor Road, Smithfield, QLD 4878 Australia
                [ ]Institute of Parasitology, University of Zurich, Winterthurerstrasse 266a, CH-8057 Zürich, Switzerland
                [ ]College of Public Health, Medical and Veterinary Sciences, James Cook University, Cairns Campus, McGregor Road, Smithfield, QLD 4878 Australia
                [ ]Department of Farm Animal, University of Zurich, Winterthurerstrasse, CH-8057 Zürich, Switzerland
                [ ]Functional Genomics Center Zurich, Winterthurerstrasse, CH-8057 Zürich, Switzerland
                [ ]Apicomplexes et Immunité Mucosale, INRA, UMR1282, Infectiologie et Santé Publique, F-37380 Nouzilly, France
                [ ]Université François Rabelais de Tours, UMR1282, Infectiologie et Santé Publique, F-37000 Tours, France
                Article
                1298
                10.1186/s12864-015-1298-6
                4345034
                25765081
                2d2d29c6-af12-464d-906b-118ec8b672b0
                © Walker et al.; licensee BioMed Central. 2015

                This is an Open Access article distributed under the terms of the Creative Commons Attribution License ( http://creativecommons.org/licenses/by/4.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly credited. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 18 September 2014
                : 29 January 2015
                Categories
                Research Article
                Custom metadata
                © The Author(s) 2015

                Genetics
                eimeria tenella,rna seq,microgametocyte,macrogametocyte,oocyst,fertilisation,transmission
                Genetics
                eimeria tenella, rna seq, microgametocyte, macrogametocyte, oocyst, fertilisation, transmission

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