Project description:We investigate changes in the single-cell transcriptomic profile of mouse (C57BL/6) splenocytes during Trypanosoma Evansi Merzoua-93 (ITMAS 150399C) infection. Infection was initiated by Intraperitoneal injection with parasites diluted in PBS. On day 0/14/42 post-injection, mice spleens were disassociated, RBC lysed to obtain the cells for 10x Chromium 3' Gene Expression Profiling
Project description:Transcriptome analysis of irradiated T evansi parasites The protozoan parasite Trypanosoma evansi is responsible for causing Surra in a variety of mammalian hosts over a wide geographical area. In order to identify which genes and processes are required to establish disease in mice, parasites were irradiated over a range using a Cobalt60 gamma source. A custom Trypanosome spp. array that covers the genomes of three trypanosome species, T. brucei, T. evansi and T. congolense was designed by Affymetrix with an average of 9300 whole gene transcripts from all three species were targeted. Irradiation differentially affected the abundance of gene transcripts in a dose-dependent trend. We present these genes as necessary for repair from irradiation damage, and essential for disease establishment in mice post irradiation.
Project description:We investigate changes in the single-cell transcriptomic profile of mouse (C57BL/6) splenocytes at late stage of Trypanosoma evansi Merzouga 93 (ITMAS 150399C) infection. Infection was initiated by Intraperitoneal injection with parasites diluted in PBS. On day 42 post-injection, mice spleens were disassociated, RBC lysed to obtain the cells for 10x Chromium 3' Gene Expression Profiling
Project description:Trypanosoma evansi is a parasite of African origin that has evolutionary proximity to Trypanosoma brucei. This parasite, which causes the surra, has the broadest range of hosts, including wild and domestic mammals, and its geographic distribution is the widest among all trypanosomes. This disease causes numerous economic losses in livestock in countries in the Americas, Africa, and Asia. This work aims to obtain in vitro secreted proteins from T. evansi and identify potential treatment and diagnostic biomarkers of surra. For this, in an experimental group, the parasite was purified in an ion-exchange column (DEAE-Cellulose) and maintained in a secretion medium. In a second group, there was no purification of the parasites. Also, each group was maintained at 37 ºC and 27 ºC. Finally, we identified the proteins through mass spectrometry. We found a total of 246 proteins and noticed that the temperature appears to modulate protein secretion. We observed negligible differences in proteins from purified and non-purified sets. We observed that most secreted proteins are related to molecular function (F) of binding, catalytic activity, structural molecule activity, and biological process (P) of metabolic process, cellular process, localization, and biological regulation. We also verified that most secretome proteins have homologous proteins with trypanosomatids T. brucei, T. b. gambiense, T. vivax, and T. equiperdum. However, despite this, it was possible to identify the presence of exclusive T. evansi epitopes with potential biomarkers for differential surra diagnosis. We identified epitopes of proteins that are part of the glycolytic process of the parasite, which can be used in immunotherapy by the production of anti-proteins from the glycolytic process. Finally, this is the first work that provide a big picture of the T. evansi secretome and could be the starting point to elucidate biological issues related to surra.
Project description:Trypanosoma evansi, the etiological agent of Surra, is the most widespread pathogenic salivarian trypanosomes and affects most livestock and wild animals in endemic regions. Knowledge of the molecular and cell biology of the parasite is limited. The study of proteins through a proteomic-based approach contributes to biochemical, molecular, and epidemiological studies. In this work, high-throughput proteomic analysis of Triton X-114-extracted surface-enriched proteins of bloodstream forms of T. evansi was carried out by gel-free approach (LC-ESI-MS/MS). Proteins resolved by SDS-PAGE revealed major proteins bands between 35 kDa and 100 kDa. After MS analysis of three biological and technical replicates, 385 proteins were identified. From these proteins, xx (0.77 %) were predicted to have GPI-anchor, 137 (35.5 %) were predicted to have transmembrane domains by (TMHMM), and 154 (40 %) were predicted as membrane-bound proteins. Membrane proteins typical of each organelle were found, some of which were already studied in other trypanosomatids. Two proteins, a and b, were considered T. evansi singletons, one of them, a, with two predicted epitopes, a putative 75 kDa invariant surface glycoprotein.
Project description:The protozoan parasite Trypanosoma evansi is responsible for causing Surra in a variety of mammalian hosts over a wide geographical area. In order to identify which genes and processes are required to establish disease in mice, parasites were irradiated over a range using a Cobalt60 gamma source. A custom Trypanosome spp. array that covers the genomes of three trypanosome species, T. brucei, T. evansi and T. congolense was designed by Affymetrix with an average of 9300 whole gene transcripts from all three species were targeted. Irradiation differentially affected the abundance of gene transcripts in a dose-dependent trend. We present these genes as necessary for repair from irradiation damage, and essential for disease establishment in mice post irradiation.
Project description:The red spider mite, Tetranychus evansi, is a oligophagous specialist mite pest of Solanaceae plants. Here, we described tomato transcriptional responses to T. evansi feeding and compared them to responses to tomato-adapted and -non-adapted strains of generalist herbivorous spider mite Tetranychus urticae. We used microarray to assess global gene expression in Solanum lycopersicum cv. Heinz 1706 upon T. evansi attack.