The complete genome sequence of Chromobacterium violaceum reveals remarkable and exploitable bacterial adaptability

Chromobacterium violaceum is one of millions of species of free-living microorganisms that populate the soil and water in the extant areas of tropical biodiversity around the world. Its complete genome sequence reveals (i) extensive alternative pathways for energy generation, (ii) ≈500 ORFs for transport-related proteins, (iii) complex and extensive systems for stress adaptation and motility, and (iv) wide-spread utilization of quorum sensing for control of inducible systems, all of which underpin the versatility and adaptability of the organism. The genome also contains extensive but incomplete arrays of ORFs coding for proteins associated with mammalian pathogenicity, possibly involved in the occasional but often fatal cases of human C. violaceum infection. There is, in addition, a series of previously unknown but important enzymes and secondary metabolites including paraquat-inducible proteins, drug and heavy-metal-resistance proteins, multiple chitinases, and proteins for the detoxification of xenobiotics that may have biotechnological applications

Susceptibility \"in vitro\'\' of agents of the chromoblastomycosis against differs antifungals

Diversos recursos terapêuticos são utilizados para controle ou cura da cromoblastomicose, mas ainda não existe terapia padrão. Poucos são os estudos relacionados à resposta dos fungos causadores frente a antifúngicos atualmente disponíveis. Nosso objetivo foi determinar os perfis de suscetibilidade desses agentes aos diferentes antifúngicos utilizados no tratamento das micoses, através da padronização da técnica de diluição em ágar e correlacionar com o tratamento vigente para alguns pacientes. Foram estudadas 39 cepas de agentes de cromoblastomicose, sendo, 12 de coleções; 30 foram identificadas como Fonsecaea pedrosoi, 6 como Phialophora verrucosa e cada uma das outras como: Cladophialophora (Cladosporium) carrionii, Fonsecaea compacta e Rhinocladiella aquaspersa. Foram realizados estudos preeliminares para escolha das melhores condições para os testes de suscetibilidade. O meio selecionado foi o yeast nitrogen base tamponado com fosfato (YNBP). O inóculo contendo cerca de 4 x 104 células/ml foi preparado a partir de cepas com 14 dias de desenvolvimento. A incubação foi realizada a 30°C e a leitura ocorreu do 3° ao 7°dias de incubação. Os testes foram realizados com: anfotericina B nas concentrações de 0,25µg/ml a 64µg/ml, 5-fluorocitosina de 0,3µg/ml a 100µg/ml, fluconazol de 0,25µg/ml a 128µg/ml, terbinafina de 0,09µg/ml a 24µg/ml, itraconazol e cetoconazol de 0,125µg/ml a 32µg/ml. Para auxiliar na interpretação dos resultados, o estabelecimento de sensiblidade ou resistência foi baseado nos níveis séricos alcançados pelas drogas. Apenas uma cepa foi sensível no teste de CIM a anfotericina B. Os valores mais altos para anfotericina B estavam entre os pacientes que fazem uso de itraconazol. Entre as 39 cepas estudadas, 77% foram resistentes ao itraconazol no teste de CIM. Para as cepas de 4 pacientes isoladas de amostras seriadas houve aumento das C IMs e para dois deles não houve melhora clínica com itraconazol. Em um desses casos houve resistência cruzada com fluconazol e outro com cetoconazol e anfotericina B. Todas as cepas foram sensíveis ao cetoconazol, mas resistentes ao fluconazol nos testes de CIM e CFM. Apenas 7 cepas responderam a 5-fluorocitosina no teste de CIM. O cetoconazol foi a droga mais eficaz, seguido da terbinafina, apesar de algumas cepas apresentaram resistência a mesma, principalmente nas procedentes de coleções. Os dados obtidos sugerem resistência inata dos fungos em estudos a algumas drogas e adquirida para algumas cepas de pacientes em tratamento com itraconazol. Esses mecanismos devem ser melhor pesquisados, além da eficácia associação de drogas, ou outra terapia e do melhor desempenho da terbinafina nos testes.Several therapeutics resources are used to control or cure of chromoblastomycossis, but there is not stand therapy yet. Feel are the studies related with fungi agents against attual antifungals. The aim of this study was to determine susceptibility profiles this fungi against differs antifungals used in treatment of mycosis, through the standaring agar diluition thecnica and relation with treatment to some pacients. 39 strains of agents were studied of chromoblastomycosis, 12 were collections, 30 were identified as F. pedrosoi, 6 as P. verrucosa and each one of others as: C. carrionii. F. compacta and R. aquaspersa. Previous studies were performed to choose the best conditions for susceptibilities tests. The culture medium selected was yeast nitrogen base buffering with phosphate (YNBP). The inoculum about 4x104 cels/ml. Was performed with strains with 14 days of developement. The incubation was performed of 30°C and the reading ocurred from the 3° to the 7° days of incubation. The tests were performed with amphotericin B in concentration of 0,25µg/ml to 64µg/ml, 5-flucytosin from 0,3µg/ml to 100µg/ml, fluconazole from 0,25µg/ml to 128µg/ml, terbinafine from 0,09µg/ml to 24µg/ml, itraconazole and ketoconazole from 0,125µg/ml to 32µg/ml. The interpretation of results was based in the plasma levels reached by the drugs. Only one strain was susceptible in the test of MIC for amphotericin B. The values highest for AmB were among the patients who used itraconazole. Among the 39 strains studied, 77% were resistents to the itraconazole in the test of MIC. For the strains there was increase of the MICs for the strains of four patients recovered from sequential samples and for to of them was not improvement with itraconazole. At one of these cases there was cross resistance with fluconazole and other with ketoconazole and amphotericin B. All the strains were susceptibles to ketoconazole, but resistents to the fluconazole in the MIC\'s and MFC\'s tests. Only 7 strains were sensibles to 5-FC in the MIC test. The terbinafine was the best drug, despite of some strains showed resistence, mainly in the culture collection. The results suggest intrinsic resistance of the fungi in study to some drugs and acquired resistance for some strains of the patients in treatment with itraconazole. These mechanism must be more researched efficacy association of drugs, or other therapy and of the best fulfillment of the terbinafine in the tests

Susceptibility \"in vitro\'\' of agents of the chromoblastomycosis against differs antifungals

Diversos recursos terapêuticos são utilizados para controle ou cura da cromoblastomicose, mas ainda não existe terapia padrão. Poucos são os estudos relacionados à resposta dos fungos causadores frente a antifúngicos atualmente disponíveis. Nosso objetivo foi determinar os perfis de suscetibilidade desses agentes aos diferentes antifúngicos utilizados no tratamento das micoses, através da padronização da técnica de diluição em ágar e correlacionar com o tratamento vigente para alguns pacientes. Foram estudadas 39 cepas de agentes de cromoblastomicose, sendo, 12 de coleções; 30 foram identificadas como Fonsecaea pedrosoi, 6 como Phialophora verrucosa e cada uma das outras como: Cladophialophora (Cladosporium) carrionii, Fonsecaea compacta e Rhinocladiella aquaspersa. Foram realizados estudos preeliminares para escolha das melhores condições para os testes de suscetibilidade. O meio selecionado foi o yeast nitrogen base tamponado com fosfato (YNBP). O inóculo contendo cerca de 4 x 104 células/ml foi preparado a partir de cepas com 14 dias de desenvolvimento. A incubação foi realizada a 30°C e a leitura ocorreu do 3° ao 7°dias de incubação. Os testes foram realizados com: anfotericina B nas concentrações de 0,25µg/ml a 64µg/ml, 5-fluorocitosina de 0,3µg/ml a 100µg/ml, fluconazol de 0,25µg/ml a 128µg/ml, terbinafina de 0,09µg/ml a 24µg/ml, itraconazol e cetoconazol de 0,125µg/ml a 32µg/ml. Para auxiliar na interpretação dos resultados, o estabelecimento de sensiblidade ou resistência foi baseado nos níveis séricos alcançados pelas drogas. Apenas uma cepa foi sensível no teste de CIM a anfotericina B. Os valores mais altos para anfotericina B estavam entre os pacientes que fazem uso de itraconazol. Entre as 39 cepas estudadas, 77% foram resistentes ao itraconazol no teste de CIM. Para as cepas de 4 pacientes isoladas de amostras seriadas houve aumento das C IMs e para dois deles não houve melhora clínica com itraconazol. Em um desses casos houve resistência cruzada com fluconazol e outro com cetoconazol e anfotericina B. Todas as cepas foram sensíveis ao cetoconazol, mas resistentes ao fluconazol nos testes de CIM e CFM. Apenas 7 cepas responderam a 5-fluorocitosina no teste de CIM. O cetoconazol foi a droga mais eficaz, seguido da terbinafina, apesar de algumas cepas apresentaram resistência a mesma, principalmente nas procedentes de coleções. Os dados obtidos sugerem resistência inata dos fungos em estudos a algumas drogas e adquirida para algumas cepas de pacientes em tratamento com itraconazol. Esses mecanismos devem ser melhor pesquisados, além da eficácia associação de drogas, ou outra terapia e do melhor desempenho da terbinafina nos testes.Several therapeutics resources are used to control or cure of chromoblastomycossis, but there is not stand therapy yet. Feel are the studies related with fungi agents against attual antifungals. The aim of this study was to determine susceptibility profiles this fungi against differs antifungals used in treatment of mycosis, through the standaring agar diluition thecnica and relation with treatment to some pacients. 39 strains of agents were studied of chromoblastomycosis, 12 were collections, 30 were identified as F. pedrosoi, 6 as P. verrucosa and each one of others as: C. carrionii. F. compacta and R. aquaspersa. Previous studies were performed to choose the best conditions for susceptibilities tests. The culture medium selected was yeast nitrogen base buffering with phosphate (YNBP). The inoculum about 4x104 cels/ml. Was performed with strains with 14 days of developement. The incubation was performed of 30°C and the reading ocurred from the 3° to the 7° days of incubation. The tests were performed with amphotericin B in concentration of 0,25µg/ml to 64µg/ml, 5-flucytosin from 0,3µg/ml to 100µg/ml, fluconazole from 0,25µg/ml to 128µg/ml, terbinafine from 0,09µg/ml to 24µg/ml, itraconazole and ketoconazole from 0,125µg/ml to 32µg/ml. The interpretation of results was based in the plasma levels reached by the drugs. Only one strain was susceptible in the test of MIC for amphotericin B. The values highest for AmB were among the patients who used itraconazole. Among the 39 strains studied, 77% were resistents to the itraconazole in the test of MIC. For the strains there was increase of the MICs for the strains of four patients recovered from sequential samples and for to of them was not improvement with itraconazole. At one of these cases there was cross resistance with fluconazole and other with ketoconazole and amphotericin B. All the strains were susceptibles to ketoconazole, but resistents to the fluconazole in the MIC\'s and MFC\'s tests. Only 7 strains were sensibles to 5-FC in the MIC test. The terbinafine was the best drug, despite of some strains showed resistence, mainly in the culture collection. The results suggest intrinsic resistance of the fungi in study to some drugs and acquired resistance for some strains of the patients in treatment with itraconazole. These mechanism must be more researched efficacy association of drugs, or other therapy and of the best fulfillment of the terbinafine in the tests

Phenotypic and genotypic characterization of Fonsecaea pedrosoi strains isolated from patients with chromoblastomycosis

A cromoblastomicose é uma infecção subcutânea causada por fungos demácios, como a Fonsecaea pedrosoi. O pleomorfismo e a similaridade entre gêneros e espécies, ocorrências características dessa classe de fungos, podem interferir na identificação morfológica. Vários esquemas terapêuticos para o tratamento da cromoblastomicose já foram utilizados, mas até o momento não existe uma terapia padrão. Em alguns casos, a correta identificação do agente etiológico pode auxiliar na indicação terapêutica, pois algumas espécies respondem melhor a determinados tratamentos do que outras. Com o objetivo de auxiliar o diagnóstico laboratorial da cromoblastomicose, foi desenvolvido um teste para identificação molecular e detecção rápida da Fonsecaea pedrosoi, empregando-se a metodologia da PCR-duplex com iniciadores específicos para a F. pedrosoi e universais para fungos. Esse teste mostrou-se altamente específico para a F. pedrosoi e, portanto, indicado para ser utilizado na identificação molecular desse fungo. Também com o intuito de avaliar as características fenotípicas e genotípicas de cepas de F. pedrosoi oriundas de pacientes com vários anos de infecção, 47 amostras, isoladas de modo seqüencial, de 11 pacientes foram analisadas por ERIC-PCR, REP-PCR, RFLP e sequenciamento da região ITS1-5.8S-ITS2 do complexo rDNA. Por ERIC-PCR e REP-PCR foram detectados 15 genótipos e, por RFLP, 6 genótipos. As técnicas combinadas geraram 20 genótipos. Com base nos resultados de sequenciamento, foi possível detectar dois tipos de seqüência que apresentaram 97% de similaridade sendo homólogas as seqüências de Fonsecaea pedrosoi disponíveis em bancos gênicos. De modo geral, pode-se observar que a espécie F. pedrosoi apresenta variação molecular e genética entre diferentes cepas e mesmo entre as cepas isoladas de um mesmo paciente.The chromoblastomycosis is a subcutaneous infection caused by dematiaceous fungi, such as the Fonsecaea pedrosoi. The pleomorfism and the similarity between genus and species that are characteristics of these fungi, can interfere on the morphologic identification. Several therapeutic regimens for the chromoblastomycosis treatment have already been used, but tell now there is not any standard therapy. In some cases, the correct identification of the etiologic agent can help in the treatment because some species have a better response to some treatments than to others. A test was developed to the molecular identification and quick detection of the Fonsecaea pedrosoi with the purpose of helping the laboratorial diagnoses of the chromoblastomycosis, using the PCR - duplex methodology with specific oligonucleotide primers for the Fonsecaea pedrosoi and universals for general fungi. This test is highly specific and sensible for the Fonsecaea pedrosoi and so, it is indicated for the molecular identification of this fungus. Another purpose of this study was to evaluate phenotypic and genotypic characteristics of the Fonsecaea pedrosoi strains isolated from patients with a long-term infection. For this reason 47 samples, isolated sequentially, from 11 patients were analyzed by ERIC-PCR, REP-PCR, RFLP and sequencing of the region ITS1 - 5.8S - ITS2 of the rDNA complex. Fifteen genotypes were found by ERIC-PCR and REP-PCR and six genotypes were found by RFLP, the combination of the methods showed twenty genotypes. Based on the results of sequencing it was possible to detect 2 types sequences that presented 97% of similarity and the available Fonsecaea pedrosoi sequences in gene banks were homological. In general, the Fonsecaea pedrosoi strains presents a molecular and genetic variation between different strains and even between isolated strains of the same patients

Phenotypic and genotypic characterization of Fonsecaea pedrosoi strains isolated from patients with chromoblastomycosis

A cromoblastomicose é uma infecção subcutânea causada por fungos demácios, como a Fonsecaea pedrosoi. O pleomorfismo e a similaridade entre gêneros e espécies, ocorrências características dessa classe de fungos, podem interferir na identificação morfológica. Vários esquemas terapêuticos para o tratamento da cromoblastomicose já foram utilizados, mas até o momento não existe uma terapia padrão. Em alguns casos, a correta identificação do agente etiológico pode auxiliar na indicação terapêutica, pois algumas espécies respondem melhor a determinados tratamentos do que outras. Com o objetivo de auxiliar o diagnóstico laboratorial da cromoblastomicose, foi desenvolvido um teste para identificação molecular e detecção rápida da Fonsecaea pedrosoi, empregando-se a metodologia da PCR-duplex com iniciadores específicos para a F. pedrosoi e universais para fungos. Esse teste mostrou-se altamente específico para a F. pedrosoi e, portanto, indicado para ser utilizado na identificação molecular desse fungo. Também com o intuito de avaliar as características fenotípicas e genotípicas de cepas de F. pedrosoi oriundas de pacientes com vários anos de infecção, 47 amostras, isoladas de modo seqüencial, de 11 pacientes foram analisadas por ERIC-PCR, REP-PCR, RFLP e sequenciamento da região ITS1-5.8S-ITS2 do complexo rDNA. Por ERIC-PCR e REP-PCR foram detectados 15 genótipos e, por RFLP, 6 genótipos. As técnicas combinadas geraram 20 genótipos. Com base nos resultados de sequenciamento, foi possível detectar dois tipos de seqüência que apresentaram 97% de similaridade sendo homólogas as seqüências de Fonsecaea pedrosoi disponíveis em bancos gênicos. De modo geral, pode-se observar que a espécie F. pedrosoi apresenta variação molecular e genética entre diferentes cepas e mesmo entre as cepas isoladas de um mesmo paciente.The chromoblastomycosis is a subcutaneous infection caused by dematiaceous fungi, such as the Fonsecaea pedrosoi. The pleomorfism and the similarity between genus and species that are characteristics of these fungi, can interfere on the morphologic identification. Several therapeutic regimens for the chromoblastomycosis treatment have already been used, but tell now there is not any standard therapy. In some cases, the correct identification of the etiologic agent can help in the treatment because some species have a better response to some treatments than to others. A test was developed to the molecular identification and quick detection of the Fonsecaea pedrosoi with the purpose of helping the laboratorial diagnoses of the chromoblastomycosis, using the PCR - duplex methodology with specific oligonucleotide primers for the Fonsecaea pedrosoi and universals for general fungi. This test is highly specific and sensible for the Fonsecaea pedrosoi and so, it is indicated for the molecular identification of this fungus. Another purpose of this study was to evaluate phenotypic and genotypic characteristics of the Fonsecaea pedrosoi strains isolated from patients with a long-term infection. For this reason 47 samples, isolated sequentially, from 11 patients were analyzed by ERIC-PCR, REP-PCR, RFLP and sequencing of the region ITS1 - 5.8S - ITS2 of the rDNA complex. Fifteen genotypes were found by ERIC-PCR and REP-PCR and six genotypes were found by RFLP, the combination of the methods showed twenty genotypes. Based on the results of sequencing it was possible to detect 2 types sequences that presented 97% of similarity and the available Fonsecaea pedrosoi sequences in gene banks were homological. In general, the Fonsecaea pedrosoi strains presents a molecular and genetic variation between different strains and even between isolated strains of the same patients

Significance of Aspergillus spp. isolation in defining cases of COVID-19 Associated Pulmonary Aspergillosis – CAPA

COVID-19-Associated Pulmonary Aspergillosis (CAPA) is a relatively common complication in patients with severe forms of the disease caused by the SARS-CoV-2 virus. Diagnosing and confirming CAPA is challenging. In this study, Aspergillus spp. isolation in respiratory specimens from patients with COVID-19 was evaluated for identifying cases of CAPA. In 2020‒2021, 17 Aspergillus spp. were isolated from 15 COVID-19 patients admitted to a university hospital in Brazil. Patient records were retrospectively reviewed to obtain clinical-epidemiological data and other markers of Aspergillus spp. infection and then compared with the ECMM/ISHAM criteria for defining CAPA. Probable CAPA was defined in 5/10 patients, who had Aspergillus spp. isolated from Bronchoalveolar Lavage (BAL) or a positive galactomannan blood test. Additionally, anti-Aspergillus antibodies were detected in two of these patients, during active or follow-up phases of CAPA. In another seven patients with Aspergillus spp. isolated from tracheobronchial aspirate or sputum, CAPA was presumed, mainly due to deterioration of clinical conditions and new lung imaging suggestive of fungal infection. Antifungal agents to control CAPA, particularly voriconazole, were used in 9/15 cases. In cases of probable CAPA and remaining patients, clinical conditions and comorbidities were similar, with lethality being high, at 60% and 71%, respectively. The number of CAPA cases defined by scientific criteria was lower than that assumed in the clinical context. This was largely due to the lack of BAL collection for fungal culture and the non-intensive use of other markers of invasive aspergillosis. The isolation of Aspergillus spp. in different respiratory specimens should alert clinicians to the diagnosis of CAPA

Núcleos de Ensino da Unesp: artigos 2010: volume 4: as disciplinas escolares, os temas transversais e o processo de educação

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP

Núcleos de Ensino da Unesp: artigos 2008

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq