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

ABSTRACT 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.

Virulence Profiling and Molecular Typing of Shiga Toxin-Producing E. coli (STEC) from Human Sources in Brazil.

Since no recent data characterizing Shiga toxin-producing E. coli (STEC) from human infections in Brazil are available, the present study aimed to investigate serotypes, stx genotypes, and accessory virulence genes, and also to perform pulsed-field gel electrophoresis (PFGE) and multi-locus sequence typing (MLST) of 43 STEC strains recovered from 2007 to 2017. Twenty-one distinct serotypes were found, with serotype O111:H8 being the most common. However, serotypes less frequently reported in human diseases were also found and included a hybrid STEC/ETEC O100:H25 clone. The majority of the strains carried stx1a as the sole stx genotype and were positive for the eae gene. Regarding the occurrence of 28 additional virulence genes associated with plasmids and pathogenicity islands, a diversity of profiles was found especially among the eae-harboring strains, which had combinations of markers composed of up to 12 distinct genes. Although PFGE analysis demonstrated genetic diversity between serotypes such as O157:H7, O111:H8, O26:H11, O118:H16, and O123:H2, high genetic relatedness was found for strains of serotypes O24:H4 and O145:H34. MLST allowed the identification of 17 distinct sequence types (STs) with ST 16 and 21 being the most common ones. Thirty-five percent of the strains studied were not typeable by the currently used MLST approach, suggesting new STs. Although STEC O111:H8 remains the leading serotype in Brazil, a diversity of other serotypes, some carrying virulence genes and belonging to STs incriminated as causing severe disease, were found in this study. Further studies are needed to determine whether they have any epidemiological relevance.

Chromoblastomycosis in the amazon region, BRazil, caused by fonsecaea pedrosoi, fonsecaea nubica, and Rhinocladiella similis: clinicopathology, susceptibility, and molecular identification

Chromoblastomycosis is a chronic subcutaneous disease caused by human contact with melanized fungioccurring mainly in tropical and subtropical zones worldwide. This study assessed 12 patients with chro-moblastomycosis from Rondˆonia, Brazil, Amazon region. In sum, 83.3% were men, 41.6% were from MonteNegro city, median age was 52.9 years, and median time to disease progression was 12.2 years. Lesions werelocated on the lower limbs (75%), and verruciform was prevalent form (66.6%). After 3 years of treatmentwith itraconazole, two patients were considered cured. The etiological agents were identified by the molec-ular sequence of the ribosomal internal transcribed spacer ITS1, 5.8S, and ITS2 region andß-tubulin genes.Eight strains were identified asFonsecaea pedrosoi, two wereF. nubica,and two wereRhinocladiella similis.The antifungal activity of five drugs was evaluated, and the most active drug was terbinafine (range minimalinhibitory concentration [MIC] 0.015­0.12µg/ml), itraconazole (range MIC 0.03­0.5µg/ml) and voriconazole(range MIC 0.06­0.5µg/ml). The highest MIC was 5-fluorocytosine (range MIC 2­32µg/ml), and ampho-tericin B (range MIC 0.25­2µg/ml). In conclusion, the present study expanded the epidemiological diseasedatabase and described for the first timeF. nubicaandR. similisas chromoblastomycosis agents in theBrazilian Amazon region. Our results confirmed the importance of using molecular methods to identify themelanized fungi and stimulate the recognition of the disease in other places where no cases have beenreported.

Seção de Coleção de Culturas do Instituto Adolfo Lutz – 68 Anos de história dedicados à saúde pública / Culture Collection Departmento of the Adolfo Lutz Institute – 68 years of history dedicated to public health

A Seção de Coleção de Culturas do Instituto Adolfo Lutz (IAL) foi estabelecida em 1940. O acervo foi formado com o intuito de organizar linhagens isoladas e adquiridas pelos pesquisadores, como Streptococcus spp., enviados pela Dra. Rebeca Lancefield do Instituto Rockefeller, EUA; enterobactérias, doadas pelo Dr. Luiz R. Trabulsi, procedentes do CDC, EUA, outras 507 linhagens deste mesmo centro; Salmonella spp. e Shigella spp. da Dinamarca, enviadas pelo Dr. Fritz Kauffmann; Haemophilus spp., enviadas pela Dra. Margareth Pittman, NIH, EUA, Neisseria meningitidis desta mesma instituição e outras do Centre International de Réference pour les Meningocoques, França; Mycobacterium spp., da Universidade de Osaka, Japão; cerca de 200 linhagens de Leptospira spp., da Argentina, provenientes do Instituto Pan-Americano de Proteção de Alimentos e Zoonoses (Opas); 308 do ATCC; 59 do NCTC; 14 da Colection de l'Institut Pasteur, França, e outras do INCQS, Brasil. Além das linhagens consideradas de referência, outras de importância para a saúde pública fazem parte do acervo: 336 de Neisseria meningitidis (epidemia brasileira de 1974); Haemophilus aegyptius (diagnóstico da febre purpúrica brasileira, 1986); linhagens de Vibrio causador da pandemia de cólera (Peru, 1991). Além de linhagens de protozoários: Leishmania sp e Trypanosoma sp e uma coleção de 120 fungos demácios. Atualmente, o acervo conta com 2.450 linhagens que são disponibilizadas para a comunidade científica (pesquisa de vacinas, anticorpos monoclonais), diagnósticos (produção do antígeno de Montenegro pela Seção, até 2005, diagnóstico de doença de Chagas, leptospirose e bartonelose) e também para uso industrial (controle biológico de insumos e produtos). Assim, o acervo da coleção de culturas do IAL constitui um banco genético de extrema importância para a área médica e industrial, além de fazer parte fundamental da história da saúde pública do Brasil.

Caracterização fenotípica e genotípica de cepas de Fonsecaea pedrosoi isoladas de pacientes com cromoblastomicose / 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...

Caracterização fenotípica e genotípica de cepas de Fonsecaea pedrosoi isoladas de pacientes com cromoblastomicose / 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...(AU)