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In vivo and in vitro expression of five genes involved in Corynebacterium pseudotuberculosis virulence

Caseous lymphadenitis (LC) is a chronic contagious disease caused by Corynebacterium pseudotuberculosis, which mainly affects goats and sheep. Vaccination is an effective but not yet well-established method, partly due to a lack of knowledge surrounding the most effective immunoprotective components...

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Autores principales: Corrêa, Jefferson Ivan, Stocker, Andreas, Trindade, Soraya Castro, Vale, Vera, Brito, Thais, Bastos, Bruno, Raynal, José Tadeu, de Miranda, Patrícia Mares, de Alcantara, Adriano Costa, Freire, Songeli Menezes, Costa, Lília Moura, Meyer, Roberto
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Springer Berlin Heidelberg 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5976562/
https://www.ncbi.nlm.nih.gov/pubmed/29846821
http://dx.doi.org/10.1186/s13568-018-0598-z
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author Corrêa, Jefferson Ivan
Stocker, Andreas
Trindade, Soraya Castro
Vale, Vera
Brito, Thais
Bastos, Bruno
Raynal, José Tadeu
de Miranda, Patrícia Mares
de Alcantara, Adriano Costa
Freire, Songeli Menezes
Costa, Lília Moura
Meyer, Roberto
author_facet Corrêa, Jefferson Ivan
Stocker, Andreas
Trindade, Soraya Castro
Vale, Vera
Brito, Thais
Bastos, Bruno
Raynal, José Tadeu
de Miranda, Patrícia Mares
de Alcantara, Adriano Costa
Freire, Songeli Menezes
Costa, Lília Moura
Meyer, Roberto
author_sort Corrêa, Jefferson Ivan
collection PubMed
description Caseous lymphadenitis (LC) is a chronic contagious disease caused by Corynebacterium pseudotuberculosis, which mainly affects goats and sheep. Vaccination is an effective but not yet well-established method, partly due to a lack of knowledge surrounding the most effective immunoprotective components. The present study aimed to quantify and compare the in vivo expression of genes pld (phospholipase D), cpp (CP40), nanH (neuraminidase H), sodC (superoxide dismutase C) and spaC (adhesin) using qRT-PCR, with the respective expression in vitro. Caseous material of abscesses removed from five animals was cultured, with colonies suggestive of C. pseudotuberculosis identified. RNA extraction was performed on these samples, as well as on the respective pellets derived from liquid cultures brain heart infusion. After evaluating RNA integrity, complementary DNA was synthesized, followed by the relative quantification each of the genes of interest. Mean mRNA expression of the five genes found in abscesses and in cultures differed significantly, with respective values of: nanH 811.50 ± 198.27 and 359.35 ± 75.45 (p = 0.009); cpp 856.31 ± 385.11 and 154.54 ± 94.34 (p = 0.0039); plD 922.70 ± 450.73 and 212.41 ± 153.10 (p = 0.016); sodC 1,293.53 ± 564.75 and 223.63 ± 145.58 (p = 0.016); spaC 1,157.10 ± 525.13 and 214.26 ± 125.70 (p = 0,016). Expression was observed to be 6–8 times higher in abscesses than in cultures, Indicative that is a genetic expression of the in vitro bacterium exists, yet in vivo has a greater magnitude corroborating to one of these virulence factors in the pathogenesis of LC. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13568-018-0598-z) contains supplementary material, which is available to authorized users.
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spelling pubmed-59765622018-06-11 In vivo and in vitro expression of five genes involved in Corynebacterium pseudotuberculosis virulence Corrêa, Jefferson Ivan Stocker, Andreas Trindade, Soraya Castro Vale, Vera Brito, Thais Bastos, Bruno Raynal, José Tadeu de Miranda, Patrícia Mares de Alcantara, Adriano Costa Freire, Songeli Menezes Costa, Lília Moura Meyer, Roberto AMB Express Original Article Caseous lymphadenitis (LC) is a chronic contagious disease caused by Corynebacterium pseudotuberculosis, which mainly affects goats and sheep. Vaccination is an effective but not yet well-established method, partly due to a lack of knowledge surrounding the most effective immunoprotective components. The present study aimed to quantify and compare the in vivo expression of genes pld (phospholipase D), cpp (CP40), nanH (neuraminidase H), sodC (superoxide dismutase C) and spaC (adhesin) using qRT-PCR, with the respective expression in vitro. Caseous material of abscesses removed from five animals was cultured, with colonies suggestive of C. pseudotuberculosis identified. RNA extraction was performed on these samples, as well as on the respective pellets derived from liquid cultures brain heart infusion. After evaluating RNA integrity, complementary DNA was synthesized, followed by the relative quantification each of the genes of interest. Mean mRNA expression of the five genes found in abscesses and in cultures differed significantly, with respective values of: nanH 811.50 ± 198.27 and 359.35 ± 75.45 (p = 0.009); cpp 856.31 ± 385.11 and 154.54 ± 94.34 (p = 0.0039); plD 922.70 ± 450.73 and 212.41 ± 153.10 (p = 0.016); sodC 1,293.53 ± 564.75 and 223.63 ± 145.58 (p = 0.016); spaC 1,157.10 ± 525.13 and 214.26 ± 125.70 (p = 0,016). Expression was observed to be 6–8 times higher in abscesses than in cultures, Indicative that is a genetic expression of the in vitro bacterium exists, yet in vivo has a greater magnitude corroborating to one of these virulence factors in the pathogenesis of LC. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (10.1186/s13568-018-0598-z) contains supplementary material, which is available to authorized users. Springer Berlin Heidelberg 2018-05-30 /pmc/articles/PMC5976562/ /pubmed/29846821 http://dx.doi.org/10.1186/s13568-018-0598-z Text en © The Author(s) 2018 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
spellingShingle Original Article
Corrêa, Jefferson Ivan
Stocker, Andreas
Trindade, Soraya Castro
Vale, Vera
Brito, Thais
Bastos, Bruno
Raynal, José Tadeu
de Miranda, Patrícia Mares
de Alcantara, Adriano Costa
Freire, Songeli Menezes
Costa, Lília Moura
Meyer, Roberto
In vivo and in vitro expression of five genes involved in Corynebacterium pseudotuberculosis virulence
title In vivo and in vitro expression of five genes involved in Corynebacterium pseudotuberculosis virulence
title_full In vivo and in vitro expression of five genes involved in Corynebacterium pseudotuberculosis virulence
title_fullStr In vivo and in vitro expression of five genes involved in Corynebacterium pseudotuberculosis virulence
title_full_unstemmed In vivo and in vitro expression of five genes involved in Corynebacterium pseudotuberculosis virulence
title_short In vivo and in vitro expression of five genes involved in Corynebacterium pseudotuberculosis virulence
title_sort in vivo and in vitro expression of five genes involved in corynebacterium pseudotuberculosis virulence
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5976562/
https://www.ncbi.nlm.nih.gov/pubmed/29846821
http://dx.doi.org/10.1186/s13568-018-0598-z
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