Sporothrix

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Zoilo Pires De Camargo - One of the best experts on this subject based on the ideXlab platform.

  • Antifungal activity of deferiprone and EDTA against Sporothrix spp.: Effect on planktonic growth and biofilm formation.
    Medical Mycology, 2020
    Co-Authors: Raimunda Sâmia Nogueira Brilhante, Zoilo Pires De Camargo, Anderson Messias Rodrigues, Jose Julio Costa Sidrim, Anderson Da Cunha Costa, Vandbergue Santos Pereira, Mirele Rodrigues Fernandes, Jonathas Sales De Oliveira, Waldemiro De Aquino Pereira-neto, Marcos Fábio Gadelha Rocha
    Abstract:

    The present study evaluated the antifungal activity of the chelators deferiprone (DFP) and ethylenediaminetetraacetic acid (EDTA) and their effect on biofilm formation of the S. schenckii complex. Eighteen strains of Sporothrix spp. (seven S. brasiliensis, three S. globosa, three S. mexicana and five Sporothrix schenckii sensu stricto) were used. Minimum inhibitory concentration (MIC) values for EDTA and DFP against filamentous forms of Sporothrix spp. ranged from 32 to 128 μg/ml. For antifungal drugs, MIC values ranged from 0.25 to 4 μg/ml for amphotericin B, from 0.25 to 4 μg/ml for itraconazole, and from 0.03 to 0.25 μg/ml for terbinafine. The chelators caused inhibition of Sporothrix spp. in yeast form at concentrations ranging from 16 to 64 μg/ml (for EDTA) and 8 to 32 μg/ml (for DFP). For antifungal drugs, MIC values observed against the yeast varied from 0.03 to 0.5 μg/ml for AMB, 0.03 to 1 μg/ml for ITC, and 0.03 to 0.13 μg/ml for TRB. Both DFP and EDTA presented synergistic interaction with antifungals against Sporothrix spp. in both filamentous and yeast form. Biofilms formed in the presence of the chelators (512 μg/ml) showed a reduction of 47% in biomass and 45% in metabolic activity. Our data reveal that DFP and EDTA reduced the growth of planktonic cells of Sporothrix spp., had synergistic interaction with antifungal drugs against this pathogen, and reduced biofilm formation of Sporothrix spp. LAY SUMMARY Our data reveal that iron chelators deferiprone and ethylenediaminetetraacetic acid reduced the growth of planktonic cells of Sporothrix spp. as well as had synergistic interaction with antifungal drugs against this pathogen and reduced biofilm formation of Sporothrix spp.

  • genome wide mapping using new aflp markers to explore intraspecific variation among pathogenic Sporothrix species
    PLOS Neglected Tropical Diseases, 2020
    Co-Authors: Jamile Ambrosio De Carvalho, Zoilo Pires De Camargo, Ferry Hagen, Matthew C Fisher, Anderson Messias Rodrigues
    Abstract:

    Sporotrichosis is a chronic subcutaneous mycosis caused by Sporothrix species, of which the main aetiological agents are S. brasiliensis, S. schenckii, and S. globosa. Infection occurs after a traumatic inoculation of Sporothrix propagules in mammals’ skin and can follow either a classic route through traumatic inoculation by plant debris (e.g., S. schenckii and S. globosa) or an alternative route through zoonotic transmission from animals (e.g., S. brasiliensis). Epizootics followed by a zoonotic route occur in Brazil, with Rio de Janeiro as the epicenter of a recent cat-transmitted epidemic. DNA-based markers are needed to explore the epidemiology of these Sporothrix expansions using molecular methods. This paper reports the use of amplified-fragment-length polymorphisms (AFLP) to assess the degree of intraspecific variability among Sporothrix species. We used whole-genome sequences from Sporothrix species to generate 2,304 virtual AFLP fingerprints. In silico screening highlighted 6 primer pair combinations to be tested in vitro. The protocol was used to genotype 27 medically relevant Sporothrix. Based on the overall scored AFLP markers (97–137 fragments), the values of polymorphism information content (PIC = 0.2552–0.3113), marker index (MI = 0.002–0.0039), effective multiplex ratio (E = 17.8519–35.2222), resolving power (Rp = 33.6296–63.1852), discriminating power (D = 0.9291–0.9662), expected heterozygosity (H = 0.3003–0.3857), and mean heterozygosity (Havp = 0.0001) demonstrated the utility of these primer combinations for discriminating Sporothrix. AFLP markers revealed cryptic diversity in species previously thought to be the most prevalent clonal type, such as S. brasiliensis, responsible for cat-transmitted sporotrichosis, and S. globosa responsible for large sapronosis outbreaks in Asia. Three combinations (#3 EcoRI-FAM-GA/MseI-TT, #5 EcoRI-FAM-GA/MseI-AG, and #6 EcoRI-FAM-TA/MseI-AA) provide the best diversity indices and lowest error rates. These methods make it easier to track routes of disease transmission during epizooties and zoonosis, and our DNA fingerprint assay can be further transferred between laboratories to give insights into the ecology and evolution of pathogenic Sporothrix species and to inform management and mitigation strategies to tackle the advance of sporotrichosis.

  • Pentamidine inhibits the growth of Sporothrix schenckii complex and exhibits synergism with antifungal agents.
    Future Microbiology, 2018
    Co-Authors: Raimunda Sâmia Nogueira Brilhante, Zoilo Pires De Camargo, Anderson Messias Rodrigues, Vandbergue Santos Pereira, Jonathas Sales De Oliveira, Waldemiro De Aquino Pereira-neto, Raissa Gp Lopes, D. S. C. M. Castelo-branco, Rossana De Aguiar Cordeiro, J. J. C. Sidrim
    Abstract:

    AIM The purpose of this study was to evaluate the effects of the antileishmanials meglumine antimoniate and pentamidine against Sporothrix schenckii complex. MATERIALS & METHODS The antifungal activity of the two antileishmanials was assessed by broth microdilution. The interaction between the antileishmanials and antifungal drugs (amphotericin B, itraconazole and terbinafine) was evaluated by the checkerboard assay. The effect of prior exposure of Sporothrix spp. yeast cells to antileishmanials was evaluated by broth microdilution. RESULTS Only pentamidine showed antifungal activity against Sporothrix spp. Synergistic interactions were observed between pentamidine and the antifungals. Also, the pre-exposure to meglumine antimoniate reduced the susceptibility of Sardinella brasiliensis and S. schenckii sensu stricto to amphotericin B and itraconazole. CONCLUSION Pentamidine showed antifungal activity against Sporothrix spp., indicating it is a possible therapeutic alternative for the treatment of sporotrichosis.

  • Exploring virulence and immunogenicity in the emerging pathogen Sporothrix brasiliensis.
    PLOS Neglected Tropical Diseases, 2017
    Co-Authors: Paula Portella Della Terra, Anderson Messias Rodrigues, Geisa Ferreira Fernandes, Angela Satie Nishikaku, Eva Burger, Zoilo Pires De Camargo
    Abstract:

    Sporotrichosis is a polymorphic chronic infection of humans and animals classically acquired after traumatic inoculation with soil and plant material contaminated with Sporothrix spp. propagules. An alternative and successful route of transmission is bites and scratches from diseased cats, through which Sporothrix yeasts are inoculated into mammalian tissue. The development of a murine model of subcutaneous sporotrichosis mimicking the alternative route of transmission is essential to understanding disease pathogenesis and the development of novel therapeutic strategies. To explore the impact of horizontal transmission in animals (e.g., cat-cat) and zoonotic transmission on Sporothrix fitness, the left hind footpads of BALB/c mice were inoculated with 5×106 yeasts (n = 11 S. brasiliensis, n = 2 S. schenckii, or n = 1 S. globosa). Twenty days post-infection, our model reproduced both the pathophysiology and symptomology of sporotrichosis with suppurating subcutaneous nodules that progressed proximally along lymphatic channels. Across the main pathogenic members of the S. schenckii clade, S. brasiliensis was usually more virulent than S. schenckii and S. globosa. However, the virulence in S. brasiliensis was strain-dependent, and we demonstrated that highly virulent isolates disseminate from the left hind footpad to the liver, spleen, kidneys, lungs, heart, and brain of infected animals, inducing significant and chronic weight loss (losing up to 15% of their body weight). The weight loss correlated with host death between 2 and 16 weeks post-infection. Histopathological features included necrosis, suppurative inflammation, and polymorphonuclear and mononuclear inflammatory infiltrates. Immunoblot using specific antisera and homologous exoantigen investigated the humoral response. Antigenic profiles were isolate-specific, supporting the hypothesis that different Sporothrix species can elicit a heterogeneous humoral response over time, but cross reaction was observed between S. brasiliensis and S. schenckii proteomes. Despite great diversity in the immunoblot profiles, antibodies were mainly derived against 3-carboxymuconate cyclase, a glycoprotein oscillating between 60 and 70 kDa (gp60-gp70) and a 100-kDa molecule in nearly 100% of the assays. Thus, our data broaden the current view of virulence and immunogenicity in the Sporothrix-sporotrichosis system, substantially expanding the possibilities for comparative genomic with isolates bearing divergent virulence traits and helping uncover the molecular mechanisms and evolutionary pressures underpinning the emergence of Sporothrix virulence.

  • in vitro susceptibility of antifungal drugs against Sporothrix brasiliensis recovered from cats with sporotrichosis in brazil
    Medical Mycology, 2016
    Co-Authors: Raimunda Sâmia Nogueira Brilhante, Sandro Antonio Pereira, Isabella Dib Ferreira Gremião, Anderson Messias Rodrigues, Jose Julio Costa Sidrim, Marcos Fábio Gadelha Rocha, Tânia Maria Pacheco Schubach, Zoilo Pires De Camargo
    Abstract:

    Sporotrichosis is an important subcutaneous mycosis of humans and animals. Classically, the disease is acquired upon traumatic inoculation of Sporothrix propagules from contaminated soil and plant debris. In addition, the direct horizontal transmission of Sporothrix among animals and the resulting zoonotic infection in humans highlight an alternative and efficient rout of transmission through biting and scratching. Sporothrix brasiliensis is the most virulent species of the Sporothrix schenckii complex and is responsible for the long-lasting outbreak of feline sporotrichosis in Brazil. However, antifungal susceptibility data of animal-borne isolates is scarce. Therefore, this study evaluated the in vitro activity of amphotericin B, caspofungin, itraconazole, voriconazole, fluconazole, and ketoconazole against animal-borne isolates of S. brasiliensis. The susceptibility tests were performed through broth microdilution (M38-A2). The results show the relevant activity of itraconazole, amphotericin B, and ketoconazole against S. brasiliensis, with the following MIC ranges: 0.125-2, 0.125-4 and 0.0312-2 μg/ml, respectively. Caspofungin was moderately effective, displaying higher variation in MIC values (0.25-64 μg/ml). Voriconazole (2-64 μg/ml) and fluconazole (62.5-500 μg/ml) showed low activity against S. brasiliensis strains. This study contributed to the characterization of the in vitro antifungal susceptibility of strains of S. brasiliensis recovered from cats with sporotrichosis, which have recently been considered the main source of human infections.

Anderson Messias Rodrigues - One of the best experts on this subject based on the ideXlab platform.

  • Antifungal activity of deferiprone and EDTA against Sporothrix spp.: Effect on planktonic growth and biofilm formation.
    Medical Mycology, 2020
    Co-Authors: Raimunda Sâmia Nogueira Brilhante, Zoilo Pires De Camargo, Anderson Messias Rodrigues, Jose Julio Costa Sidrim, Anderson Da Cunha Costa, Vandbergue Santos Pereira, Mirele Rodrigues Fernandes, Jonathas Sales De Oliveira, Waldemiro De Aquino Pereira-neto, Marcos Fábio Gadelha Rocha
    Abstract:

    The present study evaluated the antifungal activity of the chelators deferiprone (DFP) and ethylenediaminetetraacetic acid (EDTA) and their effect on biofilm formation of the S. schenckii complex. Eighteen strains of Sporothrix spp. (seven S. brasiliensis, three S. globosa, three S. mexicana and five Sporothrix schenckii sensu stricto) were used. Minimum inhibitory concentration (MIC) values for EDTA and DFP against filamentous forms of Sporothrix spp. ranged from 32 to 128 μg/ml. For antifungal drugs, MIC values ranged from 0.25 to 4 μg/ml for amphotericin B, from 0.25 to 4 μg/ml for itraconazole, and from 0.03 to 0.25 μg/ml for terbinafine. The chelators caused inhibition of Sporothrix spp. in yeast form at concentrations ranging from 16 to 64 μg/ml (for EDTA) and 8 to 32 μg/ml (for DFP). For antifungal drugs, MIC values observed against the yeast varied from 0.03 to 0.5 μg/ml for AMB, 0.03 to 1 μg/ml for ITC, and 0.03 to 0.13 μg/ml for TRB. Both DFP and EDTA presented synergistic interaction with antifungals against Sporothrix spp. in both filamentous and yeast form. Biofilms formed in the presence of the chelators (512 μg/ml) showed a reduction of 47% in biomass and 45% in metabolic activity. Our data reveal that DFP and EDTA reduced the growth of planktonic cells of Sporothrix spp., had synergistic interaction with antifungal drugs against this pathogen, and reduced biofilm formation of Sporothrix spp. LAY SUMMARY Our data reveal that iron chelators deferiprone and ethylenediaminetetraacetic acid reduced the growth of planktonic cells of Sporothrix spp. as well as had synergistic interaction with antifungal drugs against this pathogen and reduced biofilm formation of Sporothrix spp.

  • genome wide mapping using new aflp markers to explore intraspecific variation among pathogenic Sporothrix species
    PLOS Neglected Tropical Diseases, 2020
    Co-Authors: Jamile Ambrosio De Carvalho, Zoilo Pires De Camargo, Ferry Hagen, Matthew C Fisher, Anderson Messias Rodrigues
    Abstract:

    Sporotrichosis is a chronic subcutaneous mycosis caused by Sporothrix species, of which the main aetiological agents are S. brasiliensis, S. schenckii, and S. globosa. Infection occurs after a traumatic inoculation of Sporothrix propagules in mammals’ skin and can follow either a classic route through traumatic inoculation by plant debris (e.g., S. schenckii and S. globosa) or an alternative route through zoonotic transmission from animals (e.g., S. brasiliensis). Epizootics followed by a zoonotic route occur in Brazil, with Rio de Janeiro as the epicenter of a recent cat-transmitted epidemic. DNA-based markers are needed to explore the epidemiology of these Sporothrix expansions using molecular methods. This paper reports the use of amplified-fragment-length polymorphisms (AFLP) to assess the degree of intraspecific variability among Sporothrix species. We used whole-genome sequences from Sporothrix species to generate 2,304 virtual AFLP fingerprints. In silico screening highlighted 6 primer pair combinations to be tested in vitro. The protocol was used to genotype 27 medically relevant Sporothrix. Based on the overall scored AFLP markers (97–137 fragments), the values of polymorphism information content (PIC = 0.2552–0.3113), marker index (MI = 0.002–0.0039), effective multiplex ratio (E = 17.8519–35.2222), resolving power (Rp = 33.6296–63.1852), discriminating power (D = 0.9291–0.9662), expected heterozygosity (H = 0.3003–0.3857), and mean heterozygosity (Havp = 0.0001) demonstrated the utility of these primer combinations for discriminating Sporothrix. AFLP markers revealed cryptic diversity in species previously thought to be the most prevalent clonal type, such as S. brasiliensis, responsible for cat-transmitted sporotrichosis, and S. globosa responsible for large sapronosis outbreaks in Asia. Three combinations (#3 EcoRI-FAM-GA/MseI-TT, #5 EcoRI-FAM-GA/MseI-AG, and #6 EcoRI-FAM-TA/MseI-AA) provide the best diversity indices and lowest error rates. These methods make it easier to track routes of disease transmission during epizooties and zoonosis, and our DNA fingerprint assay can be further transferred between laboratories to give insights into the ecology and evolution of pathogenic Sporothrix species and to inform management and mitigation strategies to tackle the advance of sporotrichosis.

  • Pentamidine inhibits the growth of Sporothrix schenckii complex and exhibits synergism with antifungal agents.
    Future Microbiology, 2018
    Co-Authors: Raimunda Sâmia Nogueira Brilhante, Zoilo Pires De Camargo, Anderson Messias Rodrigues, Vandbergue Santos Pereira, Jonathas Sales De Oliveira, Waldemiro De Aquino Pereira-neto, Raissa Gp Lopes, D. S. C. M. Castelo-branco, Rossana De Aguiar Cordeiro, J. J. C. Sidrim
    Abstract:

    AIM The purpose of this study was to evaluate the effects of the antileishmanials meglumine antimoniate and pentamidine against Sporothrix schenckii complex. MATERIALS & METHODS The antifungal activity of the two antileishmanials was assessed by broth microdilution. The interaction between the antileishmanials and antifungal drugs (amphotericin B, itraconazole and terbinafine) was evaluated by the checkerboard assay. The effect of prior exposure of Sporothrix spp. yeast cells to antileishmanials was evaluated by broth microdilution. RESULTS Only pentamidine showed antifungal activity against Sporothrix spp. Synergistic interactions were observed between pentamidine and the antifungals. Also, the pre-exposure to meglumine antimoniate reduced the susceptibility of Sardinella brasiliensis and S. schenckii sensu stricto to amphotericin B and itraconazole. CONCLUSION Pentamidine showed antifungal activity against Sporothrix spp., indicating it is a possible therapeutic alternative for the treatment of sporotrichosis.

  • Exploring virulence and immunogenicity in the emerging pathogen Sporothrix brasiliensis.
    PLOS Neglected Tropical Diseases, 2017
    Co-Authors: Paula Portella Della Terra, Anderson Messias Rodrigues, Geisa Ferreira Fernandes, Angela Satie Nishikaku, Eva Burger, Zoilo Pires De Camargo
    Abstract:

    Sporotrichosis is a polymorphic chronic infection of humans and animals classically acquired after traumatic inoculation with soil and plant material contaminated with Sporothrix spp. propagules. An alternative and successful route of transmission is bites and scratches from diseased cats, through which Sporothrix yeasts are inoculated into mammalian tissue. The development of a murine model of subcutaneous sporotrichosis mimicking the alternative route of transmission is essential to understanding disease pathogenesis and the development of novel therapeutic strategies. To explore the impact of horizontal transmission in animals (e.g., cat-cat) and zoonotic transmission on Sporothrix fitness, the left hind footpads of BALB/c mice were inoculated with 5×106 yeasts (n = 11 S. brasiliensis, n = 2 S. schenckii, or n = 1 S. globosa). Twenty days post-infection, our model reproduced both the pathophysiology and symptomology of sporotrichosis with suppurating subcutaneous nodules that progressed proximally along lymphatic channels. Across the main pathogenic members of the S. schenckii clade, S. brasiliensis was usually more virulent than S. schenckii and S. globosa. However, the virulence in S. brasiliensis was strain-dependent, and we demonstrated that highly virulent isolates disseminate from the left hind footpad to the liver, spleen, kidneys, lungs, heart, and brain of infected animals, inducing significant and chronic weight loss (losing up to 15% of their body weight). The weight loss correlated with host death between 2 and 16 weeks post-infection. Histopathological features included necrosis, suppurative inflammation, and polymorphonuclear and mononuclear inflammatory infiltrates. Immunoblot using specific antisera and homologous exoantigen investigated the humoral response. Antigenic profiles were isolate-specific, supporting the hypothesis that different Sporothrix species can elicit a heterogeneous humoral response over time, but cross reaction was observed between S. brasiliensis and S. schenckii proteomes. Despite great diversity in the immunoblot profiles, antibodies were mainly derived against 3-carboxymuconate cyclase, a glycoprotein oscillating between 60 and 70 kDa (gp60-gp70) and a 100-kDa molecule in nearly 100% of the assays. Thus, our data broaden the current view of virulence and immunogenicity in the Sporothrix-sporotrichosis system, substantially expanding the possibilities for comparative genomic with isolates bearing divergent virulence traits and helping uncover the molecular mechanisms and evolutionary pressures underpinning the emergence of Sporothrix virulence.

  • in vitro susceptibility of antifungal drugs against Sporothrix brasiliensis recovered from cats with sporotrichosis in brazil
    Medical Mycology, 2016
    Co-Authors: Raimunda Sâmia Nogueira Brilhante, Sandro Antonio Pereira, Isabella Dib Ferreira Gremião, Anderson Messias Rodrigues, Jose Julio Costa Sidrim, Marcos Fábio Gadelha Rocha, Tânia Maria Pacheco Schubach, Zoilo Pires De Camargo
    Abstract:

    Sporotrichosis is an important subcutaneous mycosis of humans and animals. Classically, the disease is acquired upon traumatic inoculation of Sporothrix propagules from contaminated soil and plant debris. In addition, the direct horizontal transmission of Sporothrix among animals and the resulting zoonotic infection in humans highlight an alternative and efficient rout of transmission through biting and scratching. Sporothrix brasiliensis is the most virulent species of the Sporothrix schenckii complex and is responsible for the long-lasting outbreak of feline sporotrichosis in Brazil. However, antifungal susceptibility data of animal-borne isolates is scarce. Therefore, this study evaluated the in vitro activity of amphotericin B, caspofungin, itraconazole, voriconazole, fluconazole, and ketoconazole against animal-borne isolates of S. brasiliensis. The susceptibility tests were performed through broth microdilution (M38-A2). The results show the relevant activity of itraconazole, amphotericin B, and ketoconazole against S. brasiliensis, with the following MIC ranges: 0.125-2, 0.125-4 and 0.0312-2 μg/ml, respectively. Caspofungin was moderately effective, displaying higher variation in MIC values (0.25-64 μg/ml). Voriconazole (2-64 μg/ml) and fluconazole (62.5-500 μg/ml) showed low activity against S. brasiliensis strains. This study contributed to the characterization of the in vitro antifungal susceptibility of strains of S. brasiliensis recovered from cats with sporotrichosis, which have recently been considered the main source of human infections.

Manoel Marques Evangelista Oliveira - One of the best experts on this subject based on the ideXlab platform.

  • Canine sporotrichosis: polyphasic taxonomy and antifungal susceptibility profiles of Sporothrix species in an endemic area in Brazil
    Brazilian Journal of Microbiology, 2020
    Co-Authors: Jéssica Sepulveda Boechat, Sandro Antonio Pereira, Ana Caroline Sá Machado, Paula Gonçalves Viana, Rodrigo Almeida-paes, Rosely Maria Zancopé-oliveira, Isabella Dib Ferreira Gremião, Manoel Marques Evangelista Oliveira
    Abstract:

    Sporotrichosis, a mycosis caused by pathogenic species of the genus Sporothrix , affects diverse species of mammals. Until 2007, Sporothrix schenckii was considered the unique etiologic agent of sporotrichosis. Canine sporotrichosis is a poorly reported disease, and the majority of cases are from Rio de Janeiro, Brazil. There are scarce studies on the characterization of canine isolates of Sporothrix schenckii complex, as well as few antifungal susceptibility data available. The aim of this study was to characterize the clinical isolates of Sporothrix from dogs from Brazil at species level and evaluate their antifungal susceptibility profile. Polyphasic taxonomy was used to characterization at species level (morphological, phenotypical characteristics, and molecular identification). Antifungal susceptibility profiles (amphotericin B, itraconazole, ketoconazole, posaconazole, and terbinafine) were determined using the Clinical and Laboratory Standards Institute broth microdilution method (M38-A2). According to phenotypic identification and molecular analysis, 46 isolates included in this study were identified as S. brasiliensis and one as S. schenckii . Amphotericin B presented the highest minimum inhibitory concentration values, and the other drugs showed effective in vitro antifungal activity. This is the first report of S. schenckii in dogs from Brazil, since S. brasiliensis is the only species that has been described in canine isolates from Rio de Janeiro to date. Nevertheless, no differences were observed in the antifungal susceptibility profiles between the S. brasiliensis and S. schenckii isolates, and it is important to continuously study new canine clinical isolates from Rio de Janeiro, Brazil.

  • Is Sporothrix chilensis circulating outside Chile
    PLOS Neglected Tropical Diseases, 2020
    Co-Authors: Carlos Alberto Tiburcio Valeriano, Rodrigo Almeida-paes, Rosely Maria Zancopé-oliveira, Vanessa Brito De Souza Rabello, Reginaldo Gonçalves De Lima-neto, Cicero Pinheiro Inácio, Ertênia Paiva Oliveira, Rejane Pereira Neves, Manoel Marques Evangelista Oliveira
    Abstract:

    Sporothrix chilensis is a mild-pathogenical specie of Sporothrix pallida complex, until now, known as restrict to Chile. Herein, we describe the first clinical isolates identified as S. chilensis in Brazil, preserved in the URM Culture Collection, by polyphasic taxonomy, and their respective antifungal profile of this emergent fungus.

  • Comparative proteomics in the three major human pathogenic species of the genus Sporothrix.
    Microbes and Infection, 2020
    Co-Authors: Mirelle Garcia Silva-bailão, Rodrigo Almeida-paes, Manoel Marques Evangelista Oliveira, Alexandre Melo Bailão, Clayton Luiz Borges, Patrícia De Sousa Lima, Luã Cardoso De Oliveira, Alexandre Siqueira Guedes Coelho, Célia Maria De Almeida Soares, Rosely Maria Zancopé-oliveira
    Abstract:

    Abstract Sporotrichosis is a subcutaneous mycosis of humans and other mammals, caused by dimorphic species of the genus Sporothrix. In Brazil, human disease is broadly linked to transmission by infected cats and is mainly caused by Sporothrix brasiliensis, Sporothrix schenckii and Sporothrix globosa. In this study, we used a nanoscale liquid chromatography coupled with tandem mass spectrometry approach to provide the yeast proteomic profiles of S. brasiliensis, S. schenckii and S. globosa. From a total of 247 identified proteins, 137 were found as differentially expressed. Functional classification revealed that most are related to carbohydrate and amino acid metabolism as well as stress response. Our data indicate that S. brasiliensis metabolism is distinct of that of S. schenckii and S. globosa, mainly regarding amino acid metabolism and cell wall remodeling, which are induced in the former. Enzymes belonging to glycolytic pathway are, on the other hand, up-regulated in S. schenckii and S. globosa. These findings may explain the previously described more virulent character of S. brasiliensis. Besides complementing genomic comparisons already published, this first comparative proteomic study provided information that indicates new aspects of Sporothrix species metabolism as well as offers information that may be useful in the development of prospective functional studies.

  • Interaction with Pantoea agglomerans Modulates Growth and Melanization of Sporothrix brasiliensis and Sporothrix schenckii
    Mycopathologia, 2019
    Co-Authors: Rodrigo Almeida-paes, Manoel Marques Evangelista Oliveira, Fábio Brito-santos, Alexandre Melo Bailão, Clayton Luiz Borges, Glauber Ribeiro De Souza Araújo, Susana Frases, Célia Maria De Almeida Soares, Rosely Maria Zancopé-oliveira
    Abstract:

    Sporothrix brasiliensis and Sporothrix schenckii stand as the most virulent agents of sporotrichosis, a worldwide-distributed subcutaneous mycosis. The origin of Sporothrix virulence seems to be associated with fungal interactions with organisms living in the same environment. To assess this hypothesis, the growth of these two species in association with Pantoea agglomerans , a bacterium with a habitat similar to Sporothrix spp., was evaluated. Growth, melanization, and gene expression of the fungus were compared in the presence or absence of the bacterium in the same culture medium. Both S. brasiliensis and S. schenckii grew in contact with P. agglomerans yielding heavily melanized conidia after 5 days of incubation at 30 °C in Sabouraud agar. This increased melanin production occurred around bacterial colonies, suggesting that fungal melanization is triggered by a diffusible bacterial product, which is also supported by a similar pattern of melanin production during Sporothrix spp. growth in contact with heat-killed P. agglomerans . Growth of P. agglomerans was similar in the presence or absence of the fungus. However, the growth of S. brasiliensis and S. schenckii was initially inhibited, but further enhanced when these species were co-cultured with P. agglomerans . Moreover, fungi were able to use killed bacteria as both carbon and nitrogen sources for growth. Representational difference analysis identified overexpressed genes related to membrane transport when S. brasiliensis was co-cultured with the bacteria. The down-regulation of metabolism-related genes appears to be related to nutrient availability during bacterial exploitation. These findings can lead to a better knowledge on Sporothrix ecology and virulence.

  • Melanins Protect Sporothrix brasiliensis and Sporothrix schenckii from the Antifungal Effects of Terbinafine.
    PLOS ONE, 2016
    Co-Authors: Rodrigo Almeida-paes, Manoel Marques Evangelista Oliveira, Maria Helena Galdino Figueiredo-carvalho, Fábio Brito-santos, Fernando Almeida-silva, Rosely Maria Zancopé-oliveira
    Abstract:

    Terbinafine is a recommended therapeutic alternative for patients with sporotrichosis who cannot use itraconazole due to drug interactions or side effects. Melanins are involved in resistance to antifungal drugs and Sporothrix species produce three different types of melanin. Therefore, in this study we evaluated whether Sporothrix melanins impact the efficacy of antifungal drugs. Minimal inhibitory concentrations (MIC) and minimal fungicidal concentrations (MFC) of two Sporothrix brasiliensis and four Sporothrix schenckii strains grown in the presence of the melanin precursors L-DOPA and L-tyrosine were similar to the MIC determined by the CLSI standard protocol for S. schenckii susceptibility to amphotericin B, ketoconazole, itraconazole or terbinafine. When MICs were determined in the presence of inhibitors to three pathways of melanin synthesis, we observed, in four strains, an increase in terbinafine susceptibility in the presence of tricyclazole, a DHN-melanin inhibitor. In addition, one S. schenckii strain grown in the presence of L-DOPA had a higher MFC value when compared to the control. Growth curves in presence of 2×MIC concentrations of terbinafine showed that pyomelanin and, to a lesser extent, eumelanin were able to protect the fungi against the fungicidal effect of this antifungal drug. Our results suggest that melanin protects the major pathogenic species of the Sporothrix complex from the effects of terbinafine and that the development of new antifungal drugs targeting melanin synthesis may improve sporotrichosis therapies.

Michael J. Wingfield - One of the best experts on this subject based on the ideXlab platform.

  • the divorce of Sporothrix and ophiostoma solution to a problematic relationship
    Studies in Mycology, 2016
    Co-Authors: Z. Wilhelm Beer, Tuan A. Duong, Michael J. Wingfield
    Abstract:

    One of the causal agents of human sporotrichosis, Sporothrix schenckii, is the type species of the genus Sporothrix. During the course of the last century the asexual morphs of many Ophiostoma spp. have also been treated in Sporothrix. More recently several DNA-based studies have suggested that species of Sporothrix and Ophiostoma converge in what has become known as Ophiostoma s. lat. Were the one fungus one name principles adopted in the Melbourne Code to be applied to Ophiostoma s. lat., Sporothrix would have priority over Ophiostoma, resulting in more than 100 new combinations. The consequence would be name changes for several economically important tree pathogens including O. novo-ulmi. Alternatively, Ophiostoma could be conserved against Sporothrix, but this would necessitate changing the names of the important human pathogens in the group. In this study, we sought to resolve the phylogenetic relationship between Ophiostoma and Sporothrix. DNA sequences were determined for the ribosomal large subunit and internal transcribed spacer regions, as well as the beta-tubulin and calmodulin genes in 65 isolates. The results revealed Sporothrix as a well-supported monophyletic lineage including 51 taxa, distinct from Ophiostoma s. str. To facilitate future studies exploring species level resolution within Sporothrix, we defined six species complexes in the genus. These include the Pathogenic Clade containing the four human pathogens, together with the S. pallida-, S. candida-, S. inflata-, S. gossypina- and S. stenoceras complexes, which include environmental species mostly from soil, hardwoods and Protea infructescences. The description of Sporothrix is emended to include sexual morphs, and 26 new combinations. Two new names are also provided for species previously treated as Ophiostoma.

  • taxonomy and phylogeny of new wood and soil inhabiting Sporothrix species in the ophiostoma stenoceras Sporothrix schenckii complex
    Mycologia, 2008
    Co-Authors: Elsie M De Meyer, Sybren G De Hoog, Wilhelm Z De Beer, Richard C Summerbell, A M Moharram, Hester F Vismer, Michael J. Wingfield
    Abstract:

    Sporothrix, one of the anamorph genera of Ophiostoma, includes the important human pathogen S. schenckii and various fungi associated with insects and sap stain of wood. A survey of fungi from wood utility poles in South Africa yielded two distinct groups of Sporothrix isolates from different geograph- ical areas. DNA sequence and morphological data derived in this study showed that isolates in these groups represent two novel species in the S. schenckii- O. stenoceras species complex. A new species isolated from pine poles and rosebush wood and phylogenet- ically closely related to S. pallida is described here as Sporothrix stylites. Phylogenetic analyses also con- firmed the synonymy of S. albicans and S. nivea with S. pallida. Sporothrix stylites and S. pallida also are related closely to the isolates from soil, previously treated as ''environmental'' isolates of S. schenckii. Soil isolates are clearly distinct from human isolates of S. schenckii. We describe the former here as Sporothrix humicola. The isolates from eucalypt poles group peripheral to most other species in the S. schenckii-O. stenoceras complex and are newly described as Sporothrix lignivora. Phylogenetic analyses of sequenc- es of isolates from soil and wood together with those of clinical isolates showed that the human-pathogenic strains form an aggregate of several cryptic species.

  • Conidium development in Sporothrix anamorphs of Ophiostoma
    Fungal Biology, 1997
    Co-Authors: E. Benade, Michael J. Wingfield
    Abstract:

    Hyalorhinocladiella and Sporothrix are two common mycelial anamorphs of Ophiostoma that are difficult to distinguish from each other. Sporothrix spp. differ visibly from Hyalorhinocladiella by the presence of denticles on the conidiogenous cells. Graphium and Sporothrix are often synanamorphs of the same Ophiostoma species and the aim of this study was to determine whether a relationship exists between sympodial conidium development in Sporothrix and annellidic development in Graphium. Conidium development was examined in Sporothrix schenckii, Ophiostoma nigrocarpum and Ophiostoma piceae. Using fluorescence microscopy, as well as scanning and transmission electron microscopy, distinct denticles were observed on the conidiogenous cells in Sporothrix spp. In some cases, these conidiogenous cells were reduced, giving them a Hyalorhinocladiella-like appearance. Results of this study suggest that a continuum in patterns of conidium development exists between Sporothrix, Hyalorhinocladiella and Graphium. The linear extent, and the angle of the proliferation stage with reference to the long axis of the conidiogenous cell, appear to determine the form of conidium development.

  • Ophiostoma protearum sp. nov. associated with Protea caffra infructescences
    Botany, 1997
    Co-Authors: Gert Van R. Marais, Michael J. Wingfield
    Abstract:

    A new ophiostomatoid fungus, Ophiostoma protearum, and its Sporothrix anamorph, is described based on isolations from infructescences of Protea caffra. The species is compared with ophiostomatoid fungi previously described from Protea infructescences in the Western Cape Province of South Africa. Based on differences in their anamorphs, there appear to be two evolutionary lines among these fungi in this niche. Key words: Ceratocystis, Ceratocystiopsis, Ophiostoma protearum, Sporothrix, Knoxdaviesia, biogeography.

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  • Sporothrix chilensis sp nov ascomycota ophiostomatales a soil borne agent of human sporotrichosis with mild pathogenic potential to mammals
    Fungal Biology, 2016
    Co-Authors: Anderson Messias Rodrigues, Sybren G De Hoog, Geisa Ferreira Fernandes, Rodrigo Cruz Choappa, Zoilo Pires De Camargo
    Abstract:

    A combination of phylogeny, evolution, morphologies and ecologies has enabled major advances in understanding the taxonomy of Sporothrix species, including members exhibiting distinct lifestyles such as saprobes, human/animal pathogens, and insect symbionts. Phylogenetic analyses of ITS1/2 + 5.8s sequences split Sporothrix genus in two well-defined groups with dissimilar ecologies. Species embedded in the Sporothrix schenckii complex are frequently agents of human and animal sporotrichosis, and some of these are responsible for large sapronoses and zoonoses around the warmer temperate regions of the world. At the other extreme, basal saprophytic species evolved in association with decaying wood and soil, and are rarely found to cause human disease. We propose to create a new taxa, Sporothrix chilensis sp. nov., to accommodate strains collected from a clinical case of onychomycosis as well as from environmental origins in Chile. Multigene analyses based on ITS1/2 + 5.8s region, beta-tubulin, calmodulin and translation elongation factor 1α revealed that S. chilensis is a member of the Sporothrix pallida complex, and the nearest taxon is Sporothrix mexicana, a rare soil-borne species, non-pathogenic to humans. The ITS region serves as a primary barcode marker, while each one of the protein-coding loci easily recognized species boundaries providing sufficient information for species identification. A disseminated model of murine sporotrichosis revealed a mild-pathogenic potential, with lung invasion. Although S. chilensis is not a primary pathogen, accidental infection may have an impact in the immunosuppressed population. With the introduction of distinct species with similar routes of transmission but different virulence, identification of Sporothrix agents at the species level is mandatory.

  • rapid identification of emerging human pathogenic Sporothrix species with rolling circle amplification
    Frontiers in Microbiology, 2015
    Co-Authors: Anderson Messias Rodrigues, Sybren G De Hoog, Mohammad Javad Najafzadeh, Zoilo Pires De Camargo
    Abstract:

    Sporothrix infections are emerging as an important human and animal threat among otherwise healthy patients, especially in Brazil and China. Correct identification of sporotrichosis agents is beneficial for epidemiological surveillance, enabling implementation of adequate public-health policies and guiding antifungal therapy. In areas of limited resources where sporotrichosis is endemic, high-throughput detection methods that are specific and sensitive are preferred over phenotypic methods that usually result in misidentification of closely related Sporothrix species. We sought to establish rolling circle amplification (RCA) as a low-cost screening tool for species-specific identification of human-pathogenic Sporothrix. We developed six species-specific padlock probes targeting polymorphisms in the gene encoding calmodulin. BLAST-searches revealed candidate probes that were conserved intraspecifically; no significant homology with sequences from humans, mice, plants or microorganisms outside members of Sporothrix were found. The accuracy of our RCA-based assay was demonstrated through the specificity of probe-template binding to 25 S. brasiliensis, 58 S. schenckii, 5 S. globosa, 1 S. luriei, 4 S. mexicana, and 3 S. pallida samples. No cross reactivity between closely related species was evident in vitro, and padlock probes yielded 100% specificity and sensitivity down to 3 × 10(6) copies of the target sequence. RCA-based speciation matched identifications via phylogenetic analysis of the gene encoding calmodulin and the rDNA operon (kappa 1.0; 95% confidence interval 1.0-1.0), supporting its use as a reliable alternative to DNA sequencing. This method is a powerful tool for rapid identification and specific detection of medically relevant Sporothrix, and due to its robustness has potential for ecological studies.

  • genetic diversity and antifungal susceptibility profiles in causative agents of sporotrichosis
    BMC Infectious Diseases, 2014
    Co-Authors: Anderson Messias Rodrigues, Sybren G De Hoog, Debora De Cassia Pires, Raimunda Sâmia Nogueira Brihante, Jose Julio Costa Sidrim, Marcos Fabio Gadelha, Arnaldo Lopes Colombo, Zoilo Pires De Camargo
    Abstract:

    Background Sporotrichosis is a chronic subcutaneous mycosis of humans and animals, which is typically acquired by traumatic inoculation of plant material contaminated with Sporothrix propagules, or via animals, mainly felines. Sporothrix infections notably occur in outbreaks, with large epidemics currently taking place in southeastern Brazil and northeastern China. Pathogenic species include Sporothrix brasiliensis, Sporothrix schenckii s. str., Sporothrix globosa, and Sporothrix luriei, which exhibit differing geographical distribution, virulence, and resistance to antifungals. The phylogenetically remote species Sporothrix mexicana also shows a mild pathogenic potential.

  • taxonomy and phylogeny of new wood and soil inhabiting Sporothrix species in the ophiostoma stenoceras Sporothrix schenckii complex
    Mycologia, 2008
    Co-Authors: Elsie M De Meyer, Sybren G De Hoog, Wilhelm Z De Beer, Richard C Summerbell, A M Moharram, Hester F Vismer, Michael J. Wingfield
    Abstract:

    Sporothrix, one of the anamorph genera of Ophiostoma, includes the important human pathogen S. schenckii and various fungi associated with insects and sap stain of wood. A survey of fungi from wood utility poles in South Africa yielded two distinct groups of Sporothrix isolates from different geograph- ical areas. DNA sequence and morphological data derived in this study showed that isolates in these groups represent two novel species in the S. schenckii- O. stenoceras species complex. A new species isolated from pine poles and rosebush wood and phylogenet- ically closely related to S. pallida is described here as Sporothrix stylites. Phylogenetic analyses also con- firmed the synonymy of S. albicans and S. nivea with S. pallida. Sporothrix stylites and S. pallida also are related closely to the isolates from soil, previously treated as ''environmental'' isolates of S. schenckii. Soil isolates are clearly distinct from human isolates of S. schenckii. We describe the former here as Sporothrix humicola. The isolates from eucalypt poles group peripheral to most other species in the S. schenckii-O. stenoceras complex and are newly described as Sporothrix lignivora. Phylogenetic analyses of sequenc- es of isolates from soil and wood together with those of clinical isolates showed that the human-pathogenic strains form an aggregate of several cryptic species.

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