Modulation of the immune response by Fonsecaea pedrosoi morphotypes in the course of experimental chromoblastomycosis and their role on inflammatory response chronicity

A common theme across multiple fungal pathogens is their ability to impair the establishment of a protective immune response. Although early inflammation is beneficial in containing the infection, an uncontrolled inflammatory response is detrimental and may eventually oppose disease eradication. Chromoblastomycosis (CBM), a cutaneous and subcutaneous mycosis, caused by dematiaceous fungi, is capable of inducing a chronic inflammatory response. Muriform cells, the parasitic form of Fonsecaea pedrosoi, are highly prevalent in infected tissues, especially in long-standing lesions. In this study we show that hyphae and muriform cells are able to establish a murine CBM with skin lesions and histopathological aspects similar to that found in humans, with muriform cells being the most persistent fungal form, whereas mice infected with conidia do not reach the chronic phase of the disease. Moreover, in injured tissue the presence of hyphae and especially muriform cells, but not conidia, is correlated with intense production of pro-inflammatory cytokines in vivo. Highthroughput RNA sequencing analysis (RNA-Seq) performed at early time points showed a strong up-regulation of genes related to fungal recognition, cell migration, inflammation, apoptosis and phagocytosis in macrophages exposed in vitro to muriform cells, but not conidia. We also demonstrate that only muriform cells required FcγR and Dectin-1 recognition to be internalized in vitro, and this is the main fungal form responsible for the intense inflammatory pattern observed in CBM, clarifying the chronic inflammatory reaction observed in most patients. Furthermore, our findings reveal two different fungal-host interaction strategies according to fungal morphotype, highlighting fungal dimorphism as an important key in understanding the bipolar nature of inflammatory response in fungal infections

Draft genome sequence of Mycobacterium bovis 04-303, a highly virulent strain from Argentina

Mycobacterium bovis strain 04-303 was isolated from a wild boar living in a free-ranging field in Argentina. This work reports the draft genome sequence of this highly virulent strain and the genomic comparison of its major virulence-related genes with those of M. bovis strain AF2122/97 and Mycobacterium tuberculosis strain H37Rv.Instituto de BiotecnologíaFil: Nishibe, Christiane. Universidade Federal de Mato Grosso do Sul. Faculdade de Computação; BrasilFil: Canevari Castelão, Ana Beatriz. Universidade Federal de Mato Grosso do Sul. Programa de Pós-Graduação em Ciência Animal; BrasilFil: Dalla Costa, Ricardo. Life Technologies do Brasil; BrasilFil: Pinto, Beatriz Jeronimo. Life Technologies do Brasil; BrasilFil: Varuzza, Leonardo. Life Technologies do Brasil; BrasilFil: Cataldi, Angel Adrian. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; ArgentinaFil: Bernardelli, Amelia. Ceva Salud Animal; ArgentinaFil: Bigi, Fabiana. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; ArgentinaFil: Blanco, Federico Carlos. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; ArgentinaFil: Zumarraga, Martin Jose. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Biotecnología; ArgentinaFil: Almeida, Nalvo Franco. Universidade Federal de Mato Grosso do Sul. Faculdade de Computação; BrasilFil: Araujo, Flabio Ribeiro de. Empresa Brasileira de Pesquisa Agropecuária (Embrapa). Gado de Corte; Brasi

Transcriptional remodeling patterns in murine dendritic cells infected with Paracoccidioides brasiliensis : more is not necessarily better

Most people infected with the fungus Paracoccidioides spp. do not get sick, but approximately 5% develop paracoccidioidomycosis. Understanding how host immunity determinants influence disease development could lead to novel preventative or therapeutic strategies; hence, we used two mouse strains that are resistant (A/J) or susceptible (B10.A) to P. brasiliensis to study how dendritic cells (DCs) respond to the infection. RNA sequencing analysis showed that the susceptible strain DCs remodeled their transcriptomes much more intensely than those from the resistant strain, agreeing with a previous model of more intense innate immunity response in the susceptible strain. Contrastingly, these cells also repress genes/processes involved in antigen processing and presentation, such as lysosomal activity and autophagy. After the interaction with P. brasiliensis, both DCs and macrophages from the susceptible mouse reduced the autophagy marker LC3-II recruitment to the fungal phagosome compared to the resistant strain cells, confirming this pathway’s repression. These results suggest that impairment in antigen processing and presentation processes might be partially responsible for the inefficient activation of the adaptive immune response in this model

Draft genome sequences of two Mycobacterium bovis strains isolated from beef cattle in Paraguay

This work reports the draft genome sequences of the Mycobacterium bovis strains M1009 and M1010, isolated from the lymph nodes of two infected cows on a beef farm in Paraguay. Comparative genomics between these strains and other regional strains may provide more insights regarding M. bovis epidemiology in South America.Fil: Sanabria, Lidia. Servicio Nacional de Calidad y Salud Animal; ParaguayFil: Lagrave, Lorena. Servicio Nacional de Calidad y Salud Animal; ParaguayFil: Nishibe, Christiane. Universidade Federal do Mato Grosso do Sul; BrasilFil: Ribas, Augusto C.A.. Universidade Federal do Mato Grosso do Sul; BrasilFil: Zumárraga, Martín José. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas; ArgentinaFil: Almeida, Nalvo F.. Universidade Federal do Mato Grosso do Sul; BrasilFil: Araújo, Flábio R.. Embrapa Beef Cattle; Brasi

Transcriptional remodeling patterns in murine dendritic cells infected with Paracoccidioides brasiliensis: More is not necessarily better

Most people infected with the fungu

Patterns and Processes of Mycobacterium bovis Evolution Revealed by Phylogenomic Analyses

Mycobacterium bovis is an important animal pathogen worldwide, parasitizing wild and domesticated vertebrate live stocks as well as humans. We investigated evolutionary patterns and processes in 38 sequenced genomes of M. bovis from the Americas, Europe, Asia, and Africa. We found that US strains are evolving distinctly (though with low magnitude) from most South American lineages regarding core-coding size, GC-content, and codon usage bias, and we further employ agreement to this pattern as proxy for phylogenetic reliability. Three data types were used for tree inference: 1. Single copy core-coding genes (core-coding); 2. Indel blocks coded as numeric characters (indels); 3. Gene presence/absence similarly coded (pangenome). Core-coding´s performance was below expected regarded we used 2,000+ genes, but indels and pangenome individually estimated reasonable trees, with HGT rates as high as 200x the DNA substitution rate. In addressing why core-coding performed poorly, we excluded phylogenetic method bias, homologous recombination, positive, and purifying selection as possible causes, whilst transient retention of random mildly deleterious mutations due to recent diversification, and also ongoing pseudogeneization, may be plausible. Although total evidence´s retention index was as good as presence/absence data individually in identifying US samples, it additionaly found a South American clade with more lineages excluding only two Brazilian samples. We further inferred divergence times using indels + pangenome due to lack of informativeness of core-coding, recovering relatively similar date estimates to previous studies on the origin of M. bovis (< 10,000 y) regardless of the dating prior (strict vs. uncorrelated clock, uniform vs. exponential clock, different age/rate standard deviation priors), evincing the suitability and unsuspected robustness of using strictly genomic-wide presence/absence data to dating, while also decisively rejecting a strict clock for our dataset. All trees supported a relatively basal position of African strains, which were isolated from Homo sapiens, suggesting it was an important region for early diversification and confirming we were one of the earliest hosts of the species. Evolution of US strains was affected by lateral acquisition of 57 exclusive genes from, including genes important for virulence. A clade of three US localities, distant from one another, is under even faster evolutionary rates supported by core-coding, indels, and pangenome.Fil: Patané, José S.L.. Universidade de Sao Paulo; BrasilFil: Martins, Joaquim. Universidade de Sao Paulo; BrasilFil: Castelão, Ana Beatriz. Universidade Federal do Mato Grosso do Sul; BrasilFil: Nishibe, Christiane. Universidade Federal do Mato Grosso do Sul; BrasilFil: Montera, Luciana. Universidade Federal do Mato Grosso do Sul; BrasilFil: Bigi, Fabiana. Instituto Nacional de Tecnología Agropecuaria. Centro Nacional de Investigaciones Agropecuarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Zumárraga, Martín José. Instituto Nacional de Tecnología Agropecuaria. Centro Nacional de Investigaciones Agropecuarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Cataldi, Ángel Adrián. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Nacional de Investigaciones Agropecuarias; ArgentinaFil: Junior, Antônio Fonseca. Ministerio da Agricultura Pecuaria e Abastecimento de Brasil. Empresa Brasileira de Pesquisa Agropecuaria; BrasilFil: Roxo, Eliana. Instituto Biológico de São Pablo; BrasilFil: Osório, Ana Luiza A. R.. Universidade Federal do Mato Grosso do Sul; BrasilFil: Jorge, Klaudia S.. Universidade Federal do Mato Grosso do Sul; BrasilFil: Thacker, Tyler C.. United States Department of Agriculture; Estados UnidosFil: Almeida, Nalvo F.. Universidade Federal do Mato Grosso do Sul; BrasilFil: Araújo, Flabio R.. Ministerio da Agricultura Pecuaria e Abastecimento de Brasil. Empresa Brasileira de Pesquisa Agropecuaria; BrasilFil: Setubal, João C.. Universidade de Sao Paulo; Brasil. Biocomplexity Institute of Virginia Tech; Estados Unido

Patterns and Processes of Mycobacterium bovis Evolution Revealed by Phylogenomic Analyses

Mycobacterium bovis is an important animal pathogen worldwide, parasitizing wild and domesticated vertebrate live stocks as well as humans. We investigated evolutionary patterns and processes in 38 sequenced genomes of M. bovis from the Americas, Europe, Asia, and Africa. We found that US strains are evolving distinctly (though with low magnitude) from most South American lineages regarding core-coding size, GC-content, and codon usage bias, and we further employ agreement to this pattern as proxy for phylogenetic reliability. Three data types were used for tree inference: 1. Single copy core-coding genes (core-coding); 2. Indel blocks coded as numeric characters (indels); 3. Gene presence/absence similarly coded (pangenome). Core-coding´s performance was below expected regarded we used 2,000+ genes, but indels and pangenome individually estimated reasonable trees, with HGT rates as high as 200x the DNA substitution rate. In addressing why core-coding performed poorly, we excluded phylogenetic method bias, homologous recombination, positive, and purifying selection as possible causes, whilst transient retention of random mildly deleterious mutations due to recent diversification, and also ongoing pseudogeneization, may be plausible. Although total evidence´s retention index was as good as presence/absence data individually in identifying US samples, it additionaly found a South American clade with more lineages excluding only two Brazilian samples. We further inferred divergence times using indels + pangenome due to lack of informativeness of core-coding, recovering relatively similar date estimates to previous studies on the origin of M. bovis (< 10,000 y) regardless of the dating prior (strict vs. uncorrelated clock, uniform vs. exponential clock, different age/rate standard deviation priors), evincing the suitability and unsuspected robustness of using strictly genomic-wide presence/absence data to dating, while also decisively rejecting a strict clock for our dataset. All trees supported a relatively basal position of African strains, which were isolated from Homo sapiens, suggesting it was an important region for early diversification and confirming we were one of the earliest hosts of the species. Evolution of US strains was affected by lateral acquisition of 57 exclusive genes from, including genes important for virulence. A clade of three US localities, distant from one another, is under even faster evolutionary rates supported by core-coding, indels, and pangenome.Fil: Patané, José S.L.. Universidade de Sao Paulo; BrasilFil: Martins, Joaquim. Universidade de Sao Paulo; BrasilFil: Castelão, Ana Beatriz. Universidade Federal do Mato Grosso do Sul; BrasilFil: Nishibe, Christiane. Universidade Federal do Mato Grosso do Sul; BrasilFil: Montera, Luciana. Universidade Federal do Mato Grosso do Sul; BrasilFil: Bigi, Fabiana. Instituto Nacional de Tecnología Agropecuaria. Centro Nacional de Investigaciones Agropecuarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Zumárraga, Martín José. Instituto Nacional de Tecnología Agropecuaria. Centro Nacional de Investigaciones Agropecuarias; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Cataldi, Ángel Adrián. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro Nacional de Investigaciones Agropecuarias; ArgentinaFil: Junior, Antônio Fonseca. Ministerio da Agricultura Pecuaria e Abastecimento de Brasil. Empresa Brasileira de Pesquisa Agropecuaria; BrasilFil: Roxo, Eliana. Instituto Biológico de São Pablo; BrasilFil: Osório, Ana Luiza A. R.. Universidade Federal do Mato Grosso do Sul; BrasilFil: Jorge, Klaudia S.. Universidade Federal do Mato Grosso do Sul; BrasilFil: Thacker, Tyler C.. United States Department of Agriculture; Estados UnidosFil: Almeida, Nalvo F.. Universidade Federal do Mato Grosso do Sul; BrasilFil: Araújo, Flabio R.. Ministerio da Agricultura Pecuaria e Abastecimento de Brasil. Empresa Brasileira de Pesquisa Agropecuaria; BrasilFil: Setubal, João C.. Universidade de Sao Paulo; Brasil. Biocomplexity Institute of Virginia Tech; Estados Unido

Analysing nonsynonymous mutations between two Mycobacterium bovis strains with contrasting pathogenic profiles

Mycobacterium bovis (M. bovis) is the causative agent of bovine tuberculosis, a chronic infectious disease that can affect cattle, other domesticated species, wild animals and humans. This disease produces important economic losses worldwide. Two M. bovis strains (04-303 and 534) have been isolated in Argentina. Whereas the 04-303 strain was isolated from a wild boar, the 534 strain was obtained from cattle. In a previous study, six weeks after infection, the 04-303 strain induced 100% mortality in mice. By contrast, mice infected with the 534 strain survived, with limited tissue damage, after four months. In this study we compared all predictive proteins encoded in both M. bovis genomes. The comparative analysis revealed 141 polymorphic proteins between both strains. From these proteins, nine virulence proteins showed polymorphisms in 04-303, whereas five did it in the 534 strain. Remarkably, both strains contained a high level of polymorphism in proteins related to phthiocerol dimycocerosate (PDIM) synthesis or transport. Further experimental evidence indicated that only mutations in the 534 strain have an impact on PDIM synthesis. The observed reduction in PDIM content in the 534 strain, together with its low capacity to induce phagosome arrest, may be associated with the reported deficiency of this strain to replicate and survive inside bovine macrophages. The findings of this study could contribute to a better understanding of pathogenicity and virulence aspects of M. bovis, which is essential for further studies aiming at developing new vaccines and diagnostic techniques for bovines.Fil: Bigi, Maria de Las Mercedes. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales; ArgentinaFil: Vázquez, Cristina Lourdes. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas; ArgentinaFil: Castelão, Ana Beatriz C.. Universidade Federal do Mato Grosso do Sul; BrasilFil: Garcia, Elizabeth Andrea. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas; ArgentinaFil: Cataldi, Ángel Adrián. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas; ArgentinaFil: Jackson, Mary. State University of Colorado - Fort Collins; Estados UnidosFil: McNeil, Michael. State University of Colorado - Fort Collins; Estados UnidosFil: Soria, Marcelo. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales. Universidad de Buenos Aires. Facultad de Agronomía. Instituto de Investigaciones en Biociencias Agrícolas y Ambientales; ArgentinaFil: Zumárraga, Martín José. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas; ArgentinaFil: Cabruja, Matias Ezequiel. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Gago, Gabriela Marisa. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Biología Molecular y Celular de Rosario. Universidad Nacional de Rosario. Facultad de Ciencias Bioquímicas y Farmacéuticas. Instituto de Biología Molecular y Celular de Rosario; ArgentinaFil: Blanco, Federico Carlos. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas; ArgentinaFil: Nishibe, Christiane. Universidade Federal do Mato Grosso do Sul; BrasilFil: Almeida, Nalvo F. Universidade Federal do Rio Grande do Sul; BrasilFil: de Araújo, Flábio Ribeiro. Ministerio da Agricultura Pecuaria e Abastecimento de Brasil; BrasilFil: Bigi, Fabiana. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas; Argentin

Quantification of <i>F</i>. <i>pedrosoi</i> fungal cells in the tissue and production of pro-inflammatory cytokines in the course of murine CBM.

<p>Fungal cells were counted on twenty fields chosen at random in histopathological slides with the aid of a counting reticle (A-D). In all groups muriform cells (red arrows) could be identified after 15 days after infection. At the same time, hyphal fragments (brown arrow) were also present in animals infected either with hyphae (FH) (B) or muriform cells (MC) (C). Few conidia (black arrow) are observed after 15 days of infection in animals infected with <i>F</i>. <i>pedrosoi</i> conidia (FC) (A). Cell counts were expressed as cells per mm² of injured tissue. 1000x magnification (A-D). After 15 days of infection, high levels of TNF-α (E), IL-1β (F), IL-6 (G) and MCP-1 (H) were observed in groups with higher numbers of hyphae and muriform cells in the tissue. Cytokine production was measured by ELISA from homogenized footpad tissue. *P<0.05, **P<0.01 and ***P<0.001 compared to FC group.</p