Case report: First case of fungemia caused by Papiliotrema laurentii in a patient with SARS-CoV-2 infection

ABSTRACT Previously considered saprobe and non-pathogenic, the fungus Papiliotrema laurentii (formerly known as Cryptococcus laurentii), is rarely associated with human infection. Nevertheless, there has been an increase in reported infections by non-neoformans cryptococci. After a literature search on the Cochrane Library, LILACS, SciELO, MEDLINE, PubMed, and PMC (PubMed Central) databases, we conclude that this is the first case report of fungemia and probable meningitis caused by Papiliotrema laurentii in a previously immunocompetent host with associated COVID-19.

New Insights on Environmental Occurrence of Pathogenic Fungi Based on Metagenomic Data from Brazilian Cerrado Biome

Abstract: Cerrado is the second largest biome in Brazil and majorly contributes to the country's grain production. Previous studies on soil metagenomics from the Cerrado revealed an outstanding microbial diversity. In this study, the abundance of pathogenic fungi was analyzed using metagenomic sequences of the Cerrado soils under native vegetation, and under agriculture with no-tillage and conventional tillage. In total, 128,627 sequences of fungi were identified, with 43,439 representing pathogenic fungi and were distributed as follows: native 17,301 (40%), no-tillage 13,780 (32%), and conventional tillage 12,358 (28%). We identified 41 pathogenic fungal species associated with human and animal infections. The data analysis revealed that the native soils had a higher relative abundance of fungal sequences, similar to pathogenic species sequences, in relation to the total eukaryotic sequences, than the conventional tillage and no-tillage treatments, which observed a reduction in fungal abundance because of anthropogenic activities.

A Review on COVID-19 Diagnosis Tests Approved for Use in Brazil and the Impact on Pandemic Control

Abstract With the COVID-19 pandemic, many diagnostic tests (molecular or immunological) were rapidly standardised, given the urgency of the situation, many are still in the process of being validated. The main objective of this study was to review the aspects of the diagnostic kits approved in Brazil and their application in the different federative units to gather epidemiological information. In order to achieve these objectives, a survey was carried out on the data available at the regulatory agency (ANVISA) and in the literature. The main countries that have registered products in Brazil are China (51.4%), Brazil (16.6%), South Korea (9.2%), USA (8.8%) and Germany (3.6%). The methodologies of these products are based on the detection of nucleic-acid (15.8%), antigen (13%) and antibody (71.2%). In the immunological tests, it was verified that the sensitivity ranged from 55 to 100% and the specificity from 80 to 100%. The percentage of cases in the samples tested in Brazil is elevated in almost all federative units since eight states showed 40% of positive cases in tested samples, while 18 states displayed between 20 and 40%. In conclusion, this review showed that Brazil is dependent on external technology to respond to pandemics, epidemics and endemics disease and needs to improve its biotechnological scheme to solve further diseases outbreaks.

Susceptibility and molecular characterization of Candida species from patients with vulvovaginitis

Abstract Vulvovaginal candidiasis affects women of reproductive age, which represents approximately 15–25% of vaginitis cases. The present study aimed to isolate and characterize yeast from the patients irrespective of the presentation of clinical symptoms. The isolates were subjected to in vitro susceptibility profile and characterization by molecular markers, which intended to assess the distribution of species. A total of 40 isolates were obtained and identified through the CHROMagar, API20aux and by ITS and D1/D2 regions sequencing of DNAr gene. Candida albicans strains were genotyped by the ABC system and the isolates were divided into two genotypic groups. The identity of the C. albicans, C. glabrata, C. guilliermondii, C. kefyr and Saccharomyces cerevisiae isolates was confirmed by the multilocus analysis. The strains of Candida, isolated from patients with complications, were found to be resistant to nystatin but sensitive to fluconazole, amphotericin B and ketoconazole, as observed by in vitro sensitivity profile. The isolates from asymptomatic patients, i.e., the colonized group, showed a dose-dependent sensitivity to the anti-fungal agents, fluconazole and amphotericin B. However, the isolates of C. albicans that belong to distinct genotypic groups showed the same in vitro susceptibility profile.

Detection of Streptococcus mutans using padlock probe based on Rolling Circle Amplification (RCA)

The aim of this study was to develop and evaluate a padlock probe based on the Rolling Circle Amplification (RCA), which targeted to 16S-23S rDNA region of S. mutans. The specificity of developed padlock probe was tested for DNA within a panel strains, including S. mutans isolated from the saliva and reference strains of the genus Streptococcus, as well as total DNA samples of biofilm and saliva. The results were positive either for DNA samples of S. mutans or DNA samples recovered from the biofilm and saliva revealing the specificity of designed padlock probe. The padlock probe based on the RCA was proved to be an effective, reproducible method for S. mutans detection and demonstrated the possibility of a rapid detection and accurate identification of S. mutans infection.

Using molecular markers to assess Streptococcus mutans variability and the biological risk for caries

AIM: To characterize the genetic variability of Streptococcus mutans isolates and to correlate this variability with different colonization profiles observed during dental caries in a sample of children. METHODS: S. mutans samples were isolated from the saliva of 30 children with varying histories of dental caries, and they were characterized according to morphological and biochemical markers and the sequences of their 16S-23S intergenic spacer region. The genetic variability of the isolates was first assessed using Random Amplified Polymorphic DNA (RAPD) markers. Next, the isolates were differentiated by sequencing a specific region of the gene encoding the enzyme glucosyltransferase B (gtfB). RESULTS: Characterization using RAPD markers uncovered significant genetic variability among the samples and indicated the existence of clusters, which allowed us to reconstruct both the origin and clinical history of the disease. By sequencing the 16S-23S intergenic region, it was found that all of the isolates belonged to the species S. mutans. Based on the genetic similarity of the isolates and pattern of amino acid variations identified by partial sequencing of the gtfB gene, base-pair changes were identified and correlated with different virulence patterns among the isolates. CONCLUSIONS: The partial sequencing of the gtfB gene can be a useful tool for elucidating the colonization patterns of S. mutans. As amino acid variations are likely to be correlated with differences in biological risk, molecular characterization, such as that described in this paper, could be the key for assessing the development of dental caries in children...

Occurrence of sulphate reducing bacteria (SRB) associated with biocorrosion on metallic surfaces in a hydroelectric power station in Ibirama (SC) - Brazil

The aim of this study was evaluate, two methods for the detection and identification of sulphate reducing bacteria (SRB): ML medium and PCR with specific primers for SRB groups. SRB were detected through the selective medium only on carbon steel, which showed corrosion. Employing specific PCR primer, SBR were detected from all the metallic components assayed, even those that did not present visible corrosion spots, such stainless steel and copper alloys. Despite the presence or absence of corrosion at the later stages effectively by using the selective medium,, the initial stages of the corrosion could only be detected by the amplification of total DNA with SRB specific primers. The early detection of SRB could be employed for preventing the damages on metal surfaces before the installation of corrosion processes. Strategies for reducing the time spent on SRB isolation and identification could be auxiliary tools for controlling the corrosion of materials.

Estratégia multidisciplinar na prevenção e controle de doenças de manifestação clínica na infância / Multidisciplinary strategy for preventing and controlling diseases which manifest themselves clinically in early childhood

Objetivo: Apresentar os resultados do projeto multidisciplinar que consolidou um programa de manutenção de saúde a partir de procedimentos individuais e coletivos para o diagnóstico e prevenção de doenças na infância. Métodos: As atividades se pautaram na análise das condições de saúde de escolares de 6 a 12 anos, pertencentes à rede municipal de ensino da região rural de Campo Largo e metropolitana de Curitiba, Paraná, Brasil. As doenças foram investigadas por meio de anamnese, exame clínico, análise microbiológica de secreção orofaríngea e de saliva; além de exame parasitológico de fezes. Atividades interativas e educacionais apresentando informações sobre transmissão e diagnóstico de doenças como cárie, febre reumática e parasitoses foram realizadas envolvendo a comunidade estudantil e familiar. Os casos clínicos foram encaminhados para tratamento e acompanhamento. Resultados: Dentre 4 837 crianças analisadas, observou-se que 81% apresentaram alto risco de desenvolver cárie dental considerando a correlação entre concentração de Streptococcus mutans na saliva e epidemiologia da doença. Os exames copro-parasitológicos revelaram índices de positividade de 70% em crianças de área rural e de 55% em escolares de área metropolitana. Cinco crianças foram diagnosticadas como portadoras de febre reumática. Conclusão: A elevada ocorrência da doença cárie e parasitoses, bem como a existência de indivíduos portadores de febre reumática, demonstraram a demanda desta população por ações de promoção de saúde que visem à educação sanitária, a profilaxia e tratamento. Atividades de extensão desta natureza são de grande importância para as comunidades escolares assistidas.

Objective: This work was conducted to present the results of a multidisciplinary project which established a health program through individual and collective procedures for the diagnosis and prevention of childhood illnesses. Methods: The health status was evaluated of children between the ages of six and twelve in the rural district of Campo Largo and schools in metropolitan Curitiba in the state of Paraná, Brazil. Students were submitted to anamnesis and clinical examination. Samples of saliva and oropharyngeal secretions were collected for microbiological analysis. In addition, fecal exams were performed to detect intestinal parasites. Information about transmission, diagnosis and prevention of dental caries, rheumatic fever and parasites was imparted to the school community by way of lectures, flyers and play activities. All clinical cases were referred for treatment and follow-up. Results: Out of the 4 837 children analyzed, 81% were at high risk of acquiring dental caries, based on the correlation between the concentration of Streptococcus mutans in the saliva and the epidemiology of the disease. The parasitological tests showed that 70% of the children from the rural areas and 56% of those from metropolitan areas, were infected. Five children tested positive for rheumatic fever.Conclusion: The high prevalence of caries, parasitic infections and the occurrence of rheumatic fever in children point to the urgency for initiatives to promote health through actions aimed at sanitary education, prophylaxis and treatment. Educational actions of this nature are of majorimportance to the school communities attended to.

Isolamento de Cryptococcus neoformans de amostras ambientais de Curitiba e região metropolitana (Paraná, Brasil) e testes de suscetibilidade frente a drogas antifúngicas / Isolation of Cryptococcus neoformans from environmental samples of Curitiba and metropolitan region (Paraná, Brazil), and susceptibility antifungal testing

Criptococose é uma doença causada pela levedura encapsulada Cryptococcus neoformans, está intimamente ligada a pessoas imunodeprimidas, quer seja por drogas imunossupressoras administradas pós-transplantes ou por doenças imunodepressorascomo a SIDA. A doença origina-se pela penetração do fungo através das vias aéreas superiores, seguindo para os pulmões, acometendo o paciente com uma pneumonia criptocócica, com posterior disseminação do microrganismo pela via hematogênica,chegando ao sistema nervoso central e até as meninges, originando uma meningite criptocócica. As amostras analisadas neste estudo são provenientes de restos vegetais, areia de galinheiros, fezes de morcegos, restos vegetais de eucaliptos como folhas e caules, e, principalmente, fezes de pombos. A análise de 88 amostras de diferentes materiais orgânicos resultou em 11 amostras positivas para o desenvolvimento da levedura. Os 11 isolados foram testados quanto à sensibilidade frente aos antifúngicos anfotericina B, fluconazol e itraconazol. C. neoformans não apresentou resistência às drogas testadas e as concentrações mínimas inibitóriasencontradas foram: CIM de 0,03 a 1 μg/mL para anfotericina B (média de 0,515 μg/mL), CIM de 0,125 μg/mL para fluconazol e CIM de 0,03 μg/mL para itraconazol.

Cryptococcosis is a fungal disease caused by Cryptococcus neoformans is closely with the imunodepressed people, as for imunodeletion drugs managed after-transplants as mmunosuppressives diseases, the illness where originates for the inoculation of fungus through the superior airways, in the lung causes the patient with a cryptococcal pneumonia, and follow dissemination of themicroorganism for the hematogênic way arriving at the central nervous system and until meninges, originating a cryptococcal meningitis. The samples analyzed in this study are proceeding from vegetal remaining portions, sand of poulterer, excrements of bats, vegetal remaining portions of eucalyptus as leaves and stem. The analysis of 88 samples of different organic materialsresulted in 11 positive samples for the development of the yeast. The 11 isolated ones had been tested to sensitivity antifungal for amphotericin B, fluconazole and itraconazole. C. neoformans did not present resistance to the tested drugs: the minimum inhibitory concentrations was founded: MIC of 0,03 to 1 μg/mL for amphotericin B (average of 0,515 μg/mL), CIM of 0,125 μg/mL for fluconazole and CIM of 0,03 μg/mL for itraconazole.

Methodological variations in the isolation of genomic DNA from Streptococcus bacteria

In this work, genomic DNA of Streptococcus pyogenes, S. mutans and S. sobrinus was isolated using two methods: either using the detergent cetyltrimethylammonium bromide (CTAB) at 65ºC; or by applying ultrasound to a mixture of silica and celite in CTAB. The composite method that used ultrasound was the more efficient, allowing the straightforward extraction of genomic DNA from Gram-positive bacteria with good quality and reproducibility.

O gênero Streptococcus encontra-se amplamente distribuído na natureza e algumas espécies constituem a microbiota humana da cavidade bucal, como Streptococcus pyogenes, que pode estar associado a importantes doenças humanas, Streptococcus mutans e Streptococcus sobrinus, relacionados à cárie dental. O DNA genômico destas três espécies foi isolado utilizando-se dois métodos, o primeiro utilizando o detergente brometo de cetiltrimetilamônio (CTAB) à 65ºC e outro associando ultra-som a uma mistura de sílica e celite em CTAB. O método que possibilitou a extração do DNA genômico das bactérias Gram positivas, com qualidade, boa reprodutibilidade fácil execução foi aquele que utilizou ultra-som associado à sílica e celite em CTAB.

Relação entre a prevalência da doença cárie e risco microbiológico / Report Between The Caries Disease Prevalence And Microbian Risk

A cárie dentária é uma doença infecciosa, multifatorial com prevalência na infância, sendo a bactéria Streptococcus mutans principal agente etiológico, existindo uma correlação entre concentração desta na cavidade bucal e incidência da doença. Neste contexto, o trabalho objetivou relacionar incidência clínica da cárie, com concentração da bactéria na saliva de escolares, visando avaliar o risco à doença e implantar ações para seu controle. Foram realizadas avaliações de determinantes sócio-econômicos, dieta, condições clínicas e análise microbiológica da saliva. A partir da concentração de unidades formadoras de colônias (UFC/mL de saliva) em Agar Mitis Salivarius acrescido de sacarose e bacitracina; as crianças foram classificadas como alto, médio ou baixo risco de desenvolvimento à doença. Análise clínica baseada nos índices CPO-D, ceo-d e atividade de cárie foi realizada e os dados confrontados com a análise microbiológica. Um total de 237 crianças, na faixa etária de 5 a 12 anos foi avaliado. A partir dos dados clínicos, observou-se que 73% das crianças apresentaram alta prevalência de cárie, das quais 83,5% apresentaram alta concentração de S. mutans na saliva, verificando correlação bisserial por ponto, significativa entre a detecção do risco e a manifestação clínica da doença. Os resultados forneceram subsídios para ações de intervenção no processo saúde/doença.

Avaliação epidemiológica e microbiológica da doença cárie em famílias de baixa renda / Epidemiological and microbiological evaluation of caries disease in low-income families

O objetivo deste estudo foi avaliar os isolados de S. mutans da saliva em nove familias, atendidas pelo PSF de Curitiba, e compará-Ios ao padrão de desenvolvimento da doença e condições socioeconômicas. A análise microbiológica da saliva foi realizada, por estimativa de UFC/ml e bioquimismo. A análise epidemiológica foi realizada (CPO-D e ceo-d), com investigação do consumo de sacarose e higiene bucal. Detectou-se a presença S. mutans (88%) e S. sobrinus (12%). Métodos estatísticos multivariados foram utilizados, estando a cárie ativa fortemente correlacionada ao alto consumo de sacarose, alta concentração de UFC/ml de saliva e multicolonização por S. mutans. Não houve correlação entre renda e epidemiologia da doença, porém o fator escolaridade dos pais apresentou relação significativa e positiva com baixo consumo de sacarose.

The goal of the present study was to evaluate isolates of S. mutans in the saliva of nine families that were patients of the PSF of Curitiba, and to compare them according to severity of disease and socio-economic level of the family. A microbiological analysis of the saliva was conducted through an estimate of CFUs/mL, and biochemism. An epidemiological analysis was carried out (CPO-D and ceo-d), and studying saccharose consumption and mouth hygiene. S. mutans (88%) and S. sobrinus (12%) were detected. Through the multivariate analysis active caries was strongly associated with high saccharose consumption, high concentration and multicolonization by S. mutans. There was no correlation between family income and the epidemiology of the disease, although the education level of the parents was strongly associated with saccharose consumption.

Analysis of the in vitro adherence of Streptococcus mutans and Candida albicans

The objective of the present study was to investigate the in vitro adherence capacity of Streptococcus mutans and Candida albicans. Adherence assays were conducted on dental surfaces and analyzed by scanning electron microscopy (SEM). Extracted human teeth were inoculated with Streptococcus mutans or Candida albicans and with both species simultaneously, and incubated at 37ºC for 21 days. Bacterial inocula had been obtained from saliva samples of children that had been colonized by both organisms. ATCC reference strains were used as controls. SEM analyses showed that the biofilm that covered the entire analyzed dental surface was more homogeneous inoculated with the two microorganisms simultaneously than with each species separately. In a second experiment, carried out with isolates that had shown the highest adherence the isolates were tested for adherence to high-density polyethylene substrates. The potentialization of the adherence capacity of Streptococcus mutans and Candida albicans when in association was confirmed.

O objetivo deste trabalho foi investigar a capacidade de aderência in vitro de Streptococcus mutans e Candida albicans. Ensaios de aderência foram realizados in vitro na superfície dentária, com posterior análise por Microscopia Eletrônica de Varredura (M.E.V.). Dentes humanos extraídos foram inoculados com Streptococcus mutans e Candida albicans, além de ambas espécies em conjunto, e foram incubados a 37ºC por 21 dias. Os inóculos eram provenientes de amostras salivares de crianças colonizadas por ambos microrganismos. Como controles foram utilizadas linhagens de referência ATCC dos dois microrganismos. A análise por M.E.V. mostrou a formação de um biofilme que cobriu toda a superfície dentária analisada de forma mais homogênea quando incubados juntos do que separadamente. Um segundo experimento foi desenvolvido utilizando isolados mostrando maior aderência dos experimentos anteriores e cortes de polietileno de alta densidade como substrato. A potencialização da capacidade de aderência de Streptococcus mutans e Candida albicans em associação foi confirmada.

Genetic variability of Streptococcus mutans isolated from low-income families, as shown by RAPD markers

The detection of Streptococcus mutans isolates with high genetic variability in different individuals indicates the occurrence of transmissibility. In this context, nine low-income families (40 individuals in total) with similar social conditions were assessed to identify the bioserotypes of S. mutans using both biochemical and RAPD markers and to establish the degree of similarity among the intra-familial isolates. The polymorphism analysis used the coefficient of Jaccard in both a Principal Coordinates Analysis (PCO) and in a UPGMA clustering method. A total of 157 isolates were obtained from salivary samples, using the morphology of colonies recovered using MSB agar as indicators. From those, 64 were characterized biochemically as S. mutans and 10 as S. sombrinus. Genetic variability among isolates based on RAPD markers was not consistent with their intra-familial distribution. In particular, some individuals might have experienced multiple infections given the high genetic variability among their isolates. The occurrence of 4 isolates with 100 percent genetic similarity is indicative of intra-familial transmission.

A detecção de isolados de S. mutans com alta similaridade genética em indivíduos diferentes, sugere a existência de transmissibilidade. Neste contexto, nove famílias (40 indivíduos) de baixo poder aquisitivo com condições sociais homogêneas foram avaliadas, visando identificar os biosorotipos de S. mutans por meio de bioquimismo e marcadores RAPD e estabelecer o grau de similaridade entre os isolados intra-familiar. Para a análise de polimorfismo utilizou-se coeficiente de Jaccard, análise de coordenadas principais (PCO) e método "UPGMA". Foram obtidos 157 isolados, a partir de amostras salivares, usando como indicador a morfologia das colônias recuperadas em ágar MSB. Destes, 64 foram caracterizados bioquimicamente como S. mutans e 10 como S. sobrinus. A partir dos marcadores RAPD verificou-se variabilidade genética entre isolados, sendo diferente a distribuição intra-familiar destes. Em determinados indivíduos podem ter ocorrido infecções múltiplas, devido a grande variabilidade dos isolados. A existência de quatro isolados apresentando 100 por cento de similaridade, sugeriu transmissão intra-familiar.

Fatores determinantes na epidemiologia e transmissibilidade da doença cárie / Determinant factors of the epidemiology and transmissibility of the caries disease

Devido às propriedades cariogênicas do Streptococcus mutans, vários estudos têm tentado identificar a fonte e a transmissão da bactéria entre humanos. Atualmente a análise de DNA baseada em traços do genótipo oferece uma rápida e real identificação da bactéria, comparada com métodos baseados em caracterização do fenótipo. O objetivo desta revisão de literatura é demonstrar a evolução das técnicas no estudo da epidemiologia e transmissibilidade do S. mutans. Os resultados até então obtidos através de inúmeras pesquisas abrem a possibilidade de investigação de diferentes populações em situações sociais, econômicas e culturais diversas; visando maior compreensão dos mecanismos de epidemiologia e transmissão intrafamiliar dos microrganismos bucais.

Isolation of herpotrichiellacious fung from the environment

Herpotrichiellaceous fungi, common agents of chromoblastomycosis and phaeohyphomycosis, were searched in samples of rotten wood, leaf littler, bark and soil of the rhizosphere, collected in the Centro Nacional de Pesquisas de Florestas/EMBRAPA, Colombo, PR, Brazil. Morphological analyses of macro, optic and scanning electron microscopy, as well as the determination of the nutritional pattern of the isolated strains were carried out for a taxonomical study. In a total of 17 colonies, 3 isolates (17.6 per cente) were identified as species of medical relevance: Cladophialophora bantiana (Sacc.) de Hoog et al., Fonsecaea pedrosoi (Brumpt) Negroni and Phialophora verrucosa Medlar. The identifications were based on the results of the analyses and on the comparison with CBS (Centraal Bureau voor Schimmelcultures) reference strains. This investigation revealed the saprophytic existence of species known as agents of chromoblastomycosis and phaeohyphomycosis. These diseases are considered autochthonous in the studied area.(au)

Qualidade microbiológica da água utilizada em turbinas de alta rotaçäo em três condiçöes clínicas diferentes / Microbiological quality of the water used in high-speed turbines under three different clinical conditions

A qualidade microbiológica da água utilizada em turbinas de alta rotaçäo foi avaliada em equipamentos odontológicos em três condiçöes clínicas diferentes: equipamentos convencionais de clínica odontológica, cuja água provinha da rede de abastecimento público; equipamentos com sistema "flush"; equipamentos cujos reservatórios de água permitiram esterilizaçäo e abastecimento com água destilada e esterilizada. Analisou-se o grau de contaminaçäo da água usada na refrigeraçäo de brocas antes e depois da realizaçäo de procedimentos clínicos. Demonstraram-se a presença de grande quantidade de bactérias na água de abastecimento, a influência positiva do sistema de assepsia das tubulaçöes e a necessidade de adoçäo de medidas de controle da qualidade microbiana da água utilizada nos equipamentos odontológicos

Isolamento e identificaçäo de microrganismo em brinquedos utilizados em consultórios / Identification of microorganisms in toys used in pediatric dental offices

A presente pesquisa teve como objetivo reconhecer os riscos de contaminaçäo existentes em oferecer brinquedos a crianças pequenas durante consultas odontopediátricas. Para se avaliar a presença de bactérias em brinquedos utilizados em consultório odontológico, foram analisados 50 brinquedos de borracha de clínicas particulares e públicas nas cidades de Curitiba e Campinas. Os microrganismos detectados nas amostras analisadas foram bacilos gram-positivos esporulados e näo-esporulados, cocos gram-positivos (estafilococos) e bacilos gram-negativos do grupo coliformes. Esses resultados nos permitem afirmar que a prática de fornecer brinquedos durante o atendimento odontopediátrico deve ser revista, já que esses brinquedos tornam-se reservatórios de agentes infecciosos