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      Trypanosoma vivax infection in sheep: Different patterns of virulence and pathogenicity associated with differentially expressed proteomes

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          Highlights

          • Trypanosoma vivax strains exhibit different virulence and pathogenicity patterns.

          • TvMT1 strain showed low virulence and high pathogenicity.

          • TvLIEM176 strain showed high virulence and moderate pathogenicity.

          • Protein expression varies in high virulence/moderate pathogenicity strain vs low virulence/high pathogenicity strain.

          Abstract

          Cattle trypanosomosis caused by Trypanosoma vivax is a widely distributed disease in Africa and Latin America. It causes significant losses in the livestock industry and is characterized by fluctuating parasitemia, anemia, fever, lethargy, and weight loss. In this study we evaluated the virulence (capacity to multiply inside the host and to modulate the host response) and pathogenicity (ability to produce disease and/or mortality) patterns of two T. vivax strains (TvMT1 and TvLIEM176) in experimentally-infected sheep and determined the proteins differentially expressed in the proteomes of these two strains. Hematological and clinical parameters were monitored in experimentally-infected versus non-infected sheep for 60 days. All the infected animals developed discernable parasitemia at 3 days post-infection (dpi), and the first parasitemia peak was observed at 6 dpi. The maximum average value of parasitemia was 1.3 × 10 7 (95% CI, 7.9 × 10 5–2 × 10 8) parasites/ml in TvLIEM176-infected animals, and 2.5 × 10 6 (95% CI, 1.6 × 10 5–4 × 10 7) parasites/ml in TvMT1-infected ones. Anemia and clinical manifestations were more severe in the animals infected by TvMT1 strain than in those infected by TvLIEM176. In the proteomic analysis, a total of 29 proteins were identified, of which 14 exhibited significant differences in their expression levels between strains. Proteins with higher expression in TvLIEM176 were: alpha tubulin, beta tubulin, arginine kinase, glucose-regulated protein 78, paraflagellar protein 3, and T-complex protein 1 subunit theta. Proteins with higher expression in TvMT1 were: chaperonin HSP60, T-complex protein 1 subunit alpha, heat shock protein 70, pyruvate kinase, glycerol kinase, inosine-5'-monophosphate dehydrogenase, 73 kDa paraflagellar rod protein, and vacuolar ATP synthase. There was a difference in the virulence and pathogenicity between the T. vivax strains: TvLIEM176 showed high virulence and moderate pathogenicity, whereas TvMT1 showed low virulence and high pathogenicity. The proteins identified in this study are discussed for their potential involvement in strains’ virulence and pathogenicity, to be further defined as biomarkers of severity in T. vivax infections.

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          Most cited references93

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          Femtomole sequencing of proteins from polyacrylamide gels by nano-electrospray mass spectrometry.

          Molecular analysis of complex biological structures and processes increasingly requires sensitive methods for protein sequencing. Electrospray mass spectrometry has been applied to the high-sensitivity sequencing of short peptides, but technical difficulties have prevented similar success with gel-isolated proteins. Here we report a simple and robust technique for the sequencing of proteins isolated by polyacrylamide gel electrophoresis, using nano-electrospray tandem mass spectrometry. As little as 5 ng protein starting material on Coomassie- or silver-stained gels can be sequenced. Multiple-sequence stretches of up to 16 amino acids are obtained, which identify the protein unambiguously if already present in databases or provide information to clone the corresponding gene. We have applied this method to the sequencing and cloning of a protein which inhibits the proliferation of capillary endothelial cells in vitro and thus may have potential antiangiogenic effects on solid tumours.
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            A novel experimental design for comparative two-dimensional gel analysis: two-dimensional difference gel electrophoresis incorporating a pooled internal standard.

            The comparison of two-dimensional (2-D) gel images from different samples is an established method used to study differences in protein expression. Conventional methods rely on comparing images from at least 2 different gels. Due to the high variation between gels, detection and quantification of protein differences can be problematic. Two-dimensional difference gel electrophoresis (Ettan trade mark DIGE) is an emerging technique for comparative proteomics, which improves the reproducibility and reliability of differential protein expression analysis between samples. In the application of DIGE different samples are labelled with mass and charge matched spectrally resolvable fluorescent dyes and are then separated on the same 2-D gel. Using an Escherichia coli lysate "spiked" with varying amounts of four different known proteins, we have tested a novel experimental design that exploits the sample multiplexing capabilities of DIGE, by including a standard sample in each gel. The standard sample comprises equal amounts of each sample to be compared and was found to improve the accuracy of protein quantification between samples from different gels allowing accurate detection of small differences in protein levels between samples.
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              Host-pathogen interactions: the attributes of virulence.

              Virulence is one of a number of possible outcomes of host-microbe interaction. As such, microbial virulence is dependent on host factors, as exemplified by the pathogenicity of avirulent microbes in immunocompromised hosts and the lack of pathogenicity of virulent pathogens in immune hosts. Pathogen-centered views of virulence assert that pathogens are distinguished from nonpathogens by their expression of virulence factors. Although this concept appears to apply to certain microbes that cause disease in normal hosts, it does not apply to most microbes that cause disease primarily in immunocompromised hosts. The study of virulence is fraught with the paradox that virulence, despite being a microbial characteristic, can only be expressed in a susceptible host. Thus, the question "What is a pathogen?" begs the question, "What is the outcome of the host-microbe interaction?" We propose that host damage provides a common denominator that translates into the different outcomes of host-microbe interaction.
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                Author and article information

                Contributors
                Journal
                Vet Parasitol X
                Vet Parasitol X
                Veterinary Parasitology: X
                Elsevier
                2590-1389
                08 June 2019
                November 2019
                08 June 2019
                : 2
                : 100014
                Affiliations
                [a ]Laboratorio de Enfermedades Parasitarias, Facultad de Ciencias Veterinarias, Universidad del Zulia, Maracaibo, Venezuela
                [b ]Laboratorio de Inmunobiología, Centro de Estudios Biomédicos y Veterinarios, Universidad Nacional Experimental Simón Rodríguez, Caracas, Venezuela
                [c ]Laboratorio de Infectología Veterinaria, Facultad de Ciencias Veterinarias, Universidad del Zulia, Maracaibo, Venezuela
                [d ]IRD, CIRAD, Univ. Montpellier (I-MUSE), Interactions Plantes Microorganismes et Environnement (IPME), Montpellier, France
                [e ]BioCampus Montpellier, CNRS, INSERM, Univ. Montpellier, Montpellier, France
                [f ]Departamento de Sanidad Animal, Universidad de Córdoba, España
                [g ]CIRAD, UMR ASTRE, F-34398 Montpellier, France
                [h ]ASTRE, CIRAD, INRA, Univ. Montpellier (I-MUSE), Montpellier, France
                [i ]INTERTRYP, CIRAD, IRD, Univ. Montpellier (I-MUSE), Montpellier, France
                Author notes
                [* ]Corresponding author. rramirez@ 123456d.umn.edu
                [1]

                Present address: Department of Biomedical Sciences, University of Minnesota Medical School, Duluth campus, Duluth, Minnesota, USA.

                [2]

                Present address: Grupo de Investigación en Sanidad Animal y Humana, Departamento de Ciencias de la Vida, Universidad de las Fuerzas Armadas ESPE, Ecuador.

                [3]

                Present address: Department of Veterinary and Biomedical Sciences, College of Veterinary Medicine, University of Minnesota, USA.

                Article
                S2590-1389(19)30010-4 100014
                10.1016/j.vpoa.2019.100014
                7458391
                8760856a-8310-4181-80ff-d1b5175cafe0
                © 2019 The Author(s)

                This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).

                History
                : 8 February 2019
                : 22 May 2019
                : 27 May 2019
                Categories
                Article

                trypanosoma vivax,virulence,pathogenicity,proteomics,biomarkers

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