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Sporotrichosis is a neglected and emerging mycosis caused by the traumatic implantation of Sporothrix propagules into the (sub)cutaneous tissues of humans and animals. We evaluated canine sporotrichosis's clinical-therapeutic, epidemiological profile, and in vitro susceptibility of isolates to itraconazole. The variables were evaluated by a chi-square test. A total of 69 dogs were infected with Sporothrix spp., and the molecular identification revealed an overwhelming occurrence of S. brasiliensis as the etiological agent. The epidemiological profile was male (56.5%), adults (4.9 ± 1.92 years old; 69.6%), and mongrels (53.6%). The clinical signs were 76.8%, ulcers, draining tracts, and nodules were predominant, mainly in the nasal region (82.2%). Dogs were diagnosed late with an evolution time of up to 3 months (34.8%). According to the prior therapeutic information, 52.2% received empirical therapy, 79.2% antibiotics, and had a 0.29 significantly greater chance of presenting lesion evolution time ˃ 3 months (P .05; Odds Ratio [OR] 1/0.29). Additionally, 25 S. brasiliensis isolates recovered between 2006-2012 (n = 15; Minimal inhibitory concentration (MIC): 0.06-2 μg/ml) and 2013-2018 (n = 10; MIC: 2→16 μg/ml) were tested against itraconazole (ITZ). These findings highlighted the resistance to ITZ in clinical cases due to S. brasiliensis occurring after 2013, showing the temporal evolution of ITZ-resistance. We warn of the importance of accurate and early diagnosis in Sporothrix-affected areas, and we report the emergence of ITZ-resistant isolates in Southern Brazil.Sporotrichosis is a fungal zoonosis. We investigated the clinical-therapeutic, epidemiological profile, and in vitro susceptibility of isolates to itraconazole (ITZ) in canine cases in Southern Brazil. Our study highlighted the emergence of ITZ-resistant Sporothrix brasiliensis and the main challenges for clinical control of this neglected disease.

Paracoccidioidomycosis (PCM), initially reported in 1908 in the city of São Paulo, Brazil, by Adolpho Lutz, is primarily a systemic and neglected tropical mycosis that may affect individuals with certain risk factors around Latin America, especially Brazil. Paracoccidioides brasiliensis sensu stricto, a classical thermodimorphic fungus associated with PCM, was long considered to represent a monotypic taxon. However, advances in molecular taxonomy revealed several cryptic species, including Paracoccidioides americana, P. restrepiensis, P. venezuelensis, and P. lutzii, that show a preference for skin and mucous membranes, lymph nodes, and respiratory organs but can also affect many other organs. The classical diagnosis of PCM benefits from direct microscopy culture-based, biochemical, and immunological assays in a general microbiology laboratory practice providing a generic identification of the agents. However, molecular assays should be employed to identify Paracoccidioides isolates to the species level, data that would be complemented by epidemiological investigations. From a clinical perspective, all probable and confirmed cases should be treated. The choice of treatment and its duration must be considered, along with the affected organs, process severity, history of previous treatment failure, possibility of administering oral medication, associated diseases, pregnancy, and patient compliance with the proposed treatment regimen. Nevertheless, even after appropriate treatment, there may be relapses, which generally occur 5 years after the apparent cure following treatment, and also, the mycosis may be confused with other diseases. This review provides a comprehensive and critical overview of the immunopathology, laboratory diagnosis, clinical aspects, and current treatment of PCM, highlighting current issues in the identification, treatment, and patient follow-up in light of recent Paracoccidioides species taxonomic developments.

Background Paracoccidioidomycosis (PCM) is an endemic disease in Latin America. In immunocompetent hosts, PCM occurs in two main clinical forms: acute and chronic. However, in HIV-infected patients PCM may show up simultaneous manifestations of acute and chronic forms. Case report We present the case of a patient diagnosed with HIV who had disseminated skin lesions and generalized lymphadenopathy, as well as respiratory and central nervous system involvement. The PCM diagnosis was confirmed by direct KOH examination, double immunodiffusion and the isolation of the fungus in samples of an abscess in the subcostal region. The isolate was identified as Paracoccidioides brasiliensis S1 by species-specific PCR using primers for protein-coding gene GP43 (exon 2) followed by PCR-RFLP of the alpha-tubulin gene. Conclusions There are few data in literature reporting species-specific molecular identification of Paracoccidioides in HIV/PCM patients. Therefore, this case report may contribute to improve the knowledge about this severe disease, its causative cryptic species, and its consequences to patients.

Background Sporotrichosis is a subcutaneous mycosis caused by Sporothrix species that affects humans and animals. Little information on the genetic diversity and population structure of the pathogen is available for Brazil, which is needed to design effective strategies to tackle the advance of sporotrichosis in endemic areas. Objectives We assessed the genetic diversity and mating-type distribution of Sporothrix isolates recovered from human and feline cases of sporotrichosis in Espirito Santo-Brazil to better understand the population structure, epidemiology, and diversification of this pathogen, as well as to explore the possible routes of transmission involved in the ongoing outbreaks. Methods In all, 75 Sporothrix isolates were identified with phenotypic characteristics. Then, fungal DNA extraction was performed, and the species-specific PCR technique was applied, using markers directed to the calmodulin gene. The mating-type idiomorph of species was identified by PCR using primers targeting the MAT1-1 and MAT1-2 loci. Results Among the 75 Sporothrix isolates, 76% were confirmed as S. brasiliensis and 24% as S. schenckii sensu stricto. S. brasiliensis was more prevalent in the metropolitan area and S. schenckii s. str. in the mountainous region of the state. In both species, the presence of the two sexual idiomorphs was detected, suggesting that they are heterothallic species. Conclusions Our data suggest that sporotrichosis takes on an epidemic-urban character involving S. brasiliensis. This species in Espirito Santo is likely to originate from Rio de Janeiro, as most isolates harbor the same MAT 1-2 locus. We confirm that S. brasiliensis has significantly broadened its area of occurrence, an essential feature of emerging pathogens.

This study investigated the effect of the quorum sensing molecules (QSMs) farnesol, 2-phenylehtanol, tyrosol and tryptophol against planktonic cells, filamentation and biofilms of Sporothrix spp. T...

Introduction. Sporotrichosis, caused by species of the Sporothrix schenckii complex, is the most prevalent subcutaneous mycosis in many areas of Latin America. Statins are a class of drugs widely used for lowering high sterol levels through their action on 3-hydroxy-3-methylglutaryl-CoA reductase, a key enzyme in the synthesis of sterol. Aim. In this study, the antifungal activity of statins (simvastatin, atorvastatin, pravastatin) against planktonic cells and biofilms of S. schenckii complex species was evaluated, as well as the interaction of pravastatin with classical antifungals (amphotericin B, itraconazole, terbinafine). Methodology. Eighteen strains of Sporothrix species were used. The antifungal susceptibility assay was performed using the broth microdilution method. Mature biofilms were exposed to statins and metabolic activity was measured by the XTT reduction assay. Results. MICs of statins ranged from 8 to 512 μg ml−1 and from 8 to 256 μg ml−1 for filamentous and yeast forms, respectively. Regarding mature biofilms, MICs of 50 % inhibition (SMIC50) were 128 μg ml−1 for simvastatin and atorvastatin and >2048 μg ml−1 for pravastatin. MICs of 90 % inhibition (SMIC90) were 512 μg ml−1 for simvastatin and >2048 μg ml−1 for atorvastatin and pravastatin. Conclusion. These results highlight the antifungal and antibiofilm potential of statins against S. schenckii complex species.

The domestic cat is the most susceptible host to Sporothrix infection, developing severe clinical forms. Few effective antifungal agents are available for treating feline sporotrichosis, and cases of treatment failure are common. Treatment success depends on cat health status, therapy-related factors, as well as social/economic issues, but it is mainly contingent upon the host-fungus interaction. The owner's adherence is critical and should be reinforced throughout the treatment to increase the chances of a successful outcome. The antifungal agents described for feline sporotrichosis are most often used in monotherapy regimens. Due to cases in which the treatment with itraconazole failed, the use of antifungal agents in combination should be considered to achieve synergy. The combination of itraconazole and potassium iodide represents an important option for the treatment of naïve cats presenting multiple cutaneous lesions, nasal mucosal lesions and/or respiratory signs, as well as for refractory cases. However, the therapeutic options for unsuccessfully treated cases are scarce. Therefore new options are needed, even more taking into account that there are many in vitro potential molecules not available for use in cats yet. More studies are necessary to correlate in vitro antifungal susceptibility tests results and the outcome of cats treated due to sporotrichosis. This review will briefly discuss both the antifungal drugs and treatment protocols used in cats with sporotrichosis, as well as the determinants of treatment failure.

Sporothrix (order Ophiostomatales) comprises a genus with 53 species, of which S. brasiliensis, S. schenckii, S. globosa, and S. luriei cause skin infections in humans and other mammals. Remarkably, closely related Sporothrix can follow different strategies in epidemics. For example, during the cat-transmitted sporotrichosis, there is an increased prevalence of the highly virulent S. brasiliensis in South America, whereas S. schenckii and S. globosa are generally associated with a sapronotic route worldwide. Therefore, species-specific types of transmission may require distinct public health strategies to mitigate the advance of sporotrichosis, including early diagnosis, isolation of new animal cases, administration of adequate antifungal therapy, and population education on the main aspects of the disease. Here, we shed light on the system Sporothrix-sporotrichosis covering hot topics in the epidemiology and diagnosis of this important neglected disease.

Este artigo tem por objetivo analisar os componentes físicos e antrópicos da Bacia Hidrográfica do Santo Antônio (BHSA), que foram compilados os mapas bases de geologia e solos, bem como dados coletados em trabalho de campo. Esses dados forma extraídos do projeto RADAMBRASIL, da Companhia de Pesquisa em Recursos Minerais (CPRM), referente a geologia da área e Levantamento semidetalhado das classes de solo disponibilizado pelo Geoportal na escala de 1:100.000, produzido pela Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA) para os municípios que compõem a bacia. Com auxílio das técnicas de sensoriamento remoto e Sistemas de Informação Geográfica, utilizou-se dados vetoriais e matriciais para ajustar algumas classes de solos, bem como elaborar o mapa de uso da terra e cobertura vegetal com as imagens Sentinel 2A. Os resultados mostram que existem 4 formações litológicas nessa bacia, com áreas de sobreposição de camadas entre Botucatu e Serra Geral, nas áreas com relevo mais acidentado. Para geomorfologia, a BHSA apresenta um relevo diverso, com formação de planaltos (morros e morrotes) no médio curso, e platôs, no alto curso. As planícies ficam restritas os fundos de vale, e em um nível mais elevado, os terraços fluviais. Já para os solos, há um predomino de latossolos vermelho no alto curso e neossolos regolíticos e litólicos no médio curso. As formas de uso mais presentes nessa bacia foram as pastagens encontradas no médio e baixo curso e agricultura concentrada, sobretudo no alto curso. No entanto, foi identificadas áreas de agricultura em solos rasos (neossolos) que possuem alta fragilidade a erosão. As características ambientais da área de estudo revelam uma tendência para a ocorrência de processos erosivos, em decorrência da ocupação de áreas com restrições ao uso agrícola.Palavras-chave: SIGs. Sensoriamento Remoto. Uso da terra.

O presente artigo busca apresentar duas formas de entender a realidade nos dias de hoje, uma que consiste num pensamento situado e contextual, e outra que se trata de uma visão global da realidade em seus diversos setores e âmbitos. Em primeiro lugar, o pensamento pós-metafísico de Jürgen Habermas, que centralizando a análise na pragmática e na razão comunicativa, situa a questão da verdade num nível epistêmico, transcendental e dessa forma não consegue tematizar, de forma coerente e sistemática, uma teoria do mundo de forma objetiva e realista. Em segundo lugar, a Nova Metafísica de Lorenz B. Puntel, que entende a filosofia enquanto teoria universal que busca a verdade em sua investigação e na qual a semântica surge como dimensão central de articulação da unidade entre linguagem e mundo objetivo, a partir do desenvolvimento de uma nova metafísica, enquanto metafísica do Ser Primordial. Assim, este artigo pretende mostrar as diferenças entre as posições divergentes desses dois filósofos, a partir da tradição de pensamento de mais de dois mil anos que entende a filosofia enquanto teoria abrangente que trabalha as estruturas fundamentais da realidade enquanto tal.

Objetivou-se com este estudo adaptar a proposta metodológica de Ross (1994; 2012) para determinação da fragilidade potencial e ambiental da bacia hidrográfica do Rio Ivinhema. A área de estudo encontra-se integralmente dentro dos limites brasileiros, na região Centro Oeste, no Estado de Mato Grosso do Sul, fazendo fronteira com o Paraguai na sua porção Sudoeste. Foram utilizadas técnicas de geoprocessamento e os preceitos metodológicos propostos por Ross (1994 e 2012) da fragilidade potencial e ambiental, acrescentando-se novos atributos (teor de argila, matéria orgânica e grau de floculação do solo). Os resultados mostraram que cerca de 17.977 km2 ou 47,7% da área é composta por fragilidade potencial baixa, associada a solos argilosos e baixa declividade (0 – 3%). Com a inserção do uso e cobertura da terra a fragilidade ambiental alta se destacou em 16,795 km2 ou 44,6% da bacia, especialmente tendo a agricultura como maior abrangência encontrada na bacia. A inserção de novos atributos na metodologia de análise (argila, matéria orgânica e grau de floculação do solo) promoveram diferenciação significativa na elaboração do mapa final de fragilidade potencial/ambiental. Dos novos fatores inseridos o grau de floculação do solo foi a variável mais influente na configuração da fragilidade potencial média. Com o uso de tecnologias de geoprocessamento foi possível adaptar a metodologia conforme a realidade da área de estudo, permitindo o entendimento de diferentes realidades e, consequentemente, melhorar a gestão territorial. Use of physico-chemical soil variables to mapping of potential and environmental fragility in the Ivinhema river basin A B S T R A C TThis study aimed to add three new factors the methodological proposal of Ross (1994; 2012) to determine the potential and environmental fragility in the Ivinhema River basin. The study area is entirely within the Brazilian limits, in the Midwest of Mato Grosso do Sul State, bordering Paraguay in its southwestern portion. Geoprocessing techniques and the methodological precepts proposed by Ross (1994 and 2012) of potential and environmental fragility were used, adding new attributes (clay content, organic matter, and degree of soil flocculation). The results showed that about 17,977 km2 or 47.7% of the area is composed of low potential fragility, associated with clayey soils and low slopes (0 - 3%). With the insertion of land use and cover, high environmental fragility stood out in 16.795 km2 or 44.6% of the basin area, especially with agriculture as the largest scope found in the basin. The inclusion of new attributes in the analysis method (clay, organic matter, and degree of soil flocculation) promoted significant differentiation in the final map of potential/environmental fragility. The use of geoprocessing technologies made it possible to add new factors associated with potential/environmental fragility, allowing the understanding of different realities and, consequently, improving territorial management.

Sporothrix schenckii and related species are the agents of human and animal sporotrichosis. Routine diagnoses using classical mycological approaches are unspecific due to overlapping phenotypes. As the frequency and prevalence of sporotrichosis increases worldwide, developing specific, sensitive and cost-effective diagnostic tools is essential to understand the distribution patterns, map-affected areas and promote specific public health strategies to mitigate future outbreaks. Polymorphisms among the β-tubulin gene were exploited to speciate S. brasiliensis, S. schenckii and S. globosa in a one-tube multiplex probe-based qPCR assay. A panel of 84 Sporothrix revealed 100% specificity (AUC = 1.000, 95% CI = 0.971-1.000, p

Aim: This study investigated the effect of terpinen-4-ol against Sporothrix schenckii complex and its interactions with antifungals. Materials & methods: The antifungal activity of terpinen-4-ol was evaluated by broth microdilution. The potential effect on cellular ergosterol concentration was evaluated by spectrophotometry. The antibiofilm activity was evaluated by violet crystal staining and XTT reduction assay. The potential pharmacological interactions with antifungals were evaluated by the checkerboard assay. Results: terpinen-4-ol (T-OH) showed minimal inhibitory concentrations ranging from 4 to 32 mg/l decreasing cellular ergosterol content and presented a SMIC ranging from 64 to 1024 mg/l for Sporothrix spp. The combinations of T-OH with itraconazole or terbinafine were synergistic. Conclusion: T-OH has antifungal activity against Sporothrix spp. and acts synergistically with standard antifungals.

Este ensaio, conforme o título evidencia, revela brevemente as condições teóricas e práticas do reconhecimento do outro, do mundo e de si próprio, resultantes da teoria e do seu método educativo de Paulo Freire, portanto, o objetivo é refletir sobre as primeiras páginas, a introdução e o primeiro capítulo da obra reconhecidíssima, A Pedagogia do Oprimido. Longe de refletir exaustivamente essas condições, aderiu-se e estudou também o Papel da Educação na Humanização. Esse trabalho é resultado coletivo do Grupo de Pesquisa de Filosofia e Educação, paideia aristotélica e paideia tomista, atualmente, vinculado aos últimos acontecimentos políticos e educacionais do país, por isso, o artigo ganhou esse viés. Duas são fontes principais de pesquisa, a saber, obras, A Pedagogia do Oprimido (2018) e a palestra concedida no Chile em 1969, intitulada, Papel da Educação na Humanização. O que daqui resulta é o entendimento que, a libertação da vida de homens e mulheres, especificamente os oprimidos, eles/as deverão não mais hospedar seus opressores, isto é na medida em que eles/as descobrem e expulsam os opressores/as, também esses/as ficam libertos, por isso, práxis libertadora

Background: The purpose of this study was to assess the role of physical activity (PA) in muscular and functional capacity in subjects under treatment for knee osteoarthritis submitted to an interd...

O presente artigo apresenta um estudo sobre a motivação humana no ambiente de trabalho, uma vez que é de vital importância não só para a administração empresarial, bem como para todos os profissionais, tendo em vista que o "mundo do trabalho" tem passado por diversas transformações, e os trabalhadores devem estar preparados. O objetivo desse trabalho é fazer um paralelo entre teorias e "mundo real", mostrando o quanto a motivação pode ser influenciada internamente ou externamente e qual a opinião das pessoas em relação ao seu trabalho, usando como ferramentas as Teorias Motivacionais apresentadas por Maslow, Herzberg, Vroom e Lawler, na busca de obter melhores resultados.

Objectives: Historically, the Brazilian Central-West region has had high numbers of paracoccidioidomycosis (PCM) cases caused by the dimorphic fungus Paracoccidioides lutzii. Methods: This epidemiological, observational, analytical, cross-sectional study was performed to investigate the clinical and laboratory data of 44 PCM patients with a culture-proven P. lutzii infection. All patients were referred to the Systemic Mycosis Center, Júlio Muller University Hospital, Cuiabá, Brazil, during January 2017 to March 2020. The neutrophil to lymphocyte ratio (NLR) was calculated and dichotomized by its median value to include in the identification of factors associated with severity. Results: At admission, 13 (31.7%) patients showed the disseminated multifocal chronic form of PCM and 16 (36.4%) patients met the clinical severity criteria. Treatment prescribed on admission did not follow the recommendations of the Brazilian Guideline for the Clinical Management of Paracoccidioidomycosis in 26% of the severe PCM cases (prevalence ratio 0.26, 95% confidence interval 0.14–0.49; P < 0.0001). Patients with severe PCM had a higher NLR that was greater than the median (≥4.11). Conclusions: The NLR biomarker complements the criteria for PCM severity. Applying the low-cost NLR test can greatly increase the diagnostic sensitivity when screening patients for PCM and contribute to better control of the disease, management of complications, and therapeutic strategies.

In a context in which conscious development proves to be more and more necessary, the financial profitability of projects with technological and environmental efficiency is highlighted and is seen as a determining factor for growth. With that in mind, the use of energy originated by the sun, both as a source of heat and light, is conducive to supply all the energy in terms of electricity. Therefore, this research aims to develop a technical economic and financial feasibility study for the implementation of a photovoltaic system in a Private Higher Education Unit in Cajazeiras - PB. For this purpose, this research is a case study, exploratory and descriptive with a quantitative approach. The results showed that the term for the return on investment is 3.8 years and that it is expected that there will be savings of approximately R $ 2,753,000.00 over the next 25 years, which makes the installation of the system quite attractive. Thus, it is evident the benefit that the use of alternative sources can bring both to organizations and to the environment, benefiting the population as a whole. Brazil has enormous potential on the subject, which is solar energy and it can certainly be better explored by all segments of Brazilian / global industry / commerce. En un contexto en el que el desarrollo consciente resulta cada vez más necesario, se destaca la rentabilidad financiera de proyectos con eficiencia tecnológica y ambiental y se vislumbra como un factor determinante para el crecimiento. Teniendo esto en cuenta, el uso de la energía originada por el sol, tanto como fuente de calor como de luz, es propicia para suministrar toda la energía en términos de electricidad. Por tanto, esta investigación tiene como objetivo desarrollar un estudio de viabilidad técnica y económica para la implementación de un sistema fotovoltaico en una Unidad Privada de Educación Superior en Cajazeiras - PB. Para ello, esta investigación es un estudio de caso, exploratorio y descriptivo con enfoque cuantitativo. Los resultados mostraron que el período de retorno de la inversión es de 3,8 años y que se espera un ahorro de aproximadamente R $ 2.753.000,00 en los próximos 25 años, lo que hace bastante atractiva la instalación del sistema. Así, es evidente el beneficio que el uso de fuentes alternativas puede traer tanto a las organizaciones como al medio ambiente, beneficiando a la población en su conjunto. Brasil tiene un enorme potencial en el tema, que es la energía solar y ciertamente puede ser mejor explorado por todos los segmentos de la industria / comercio brasileño / global. Em um contexto em que o desenvolvimento consciente se mostra cada dia mais necessário, a rentabilidade financeira de projetos com eficiência tecnológica e ambiental ganha destaque, e é vista como fator determinante para o crescimento. Pensando nisso, o aproveitamento da energia originada pelo sol, tanto enquanto fonte de calor como de luz, é propicio para fornecer toda a energia em termos de eletricidade. Assim sendo, essa pesquisa tem como objetivo desenvolver um estudo de viabilidade técnica econômico-financeira de implantação de um sistema fotovoltaico em uma Unidade de Ensino Superior Particular em Cajazeiras – PB. Para tanto a presente pesquisa trata-se de um estudo de caso, exploratório e descritivo de abordagem quantitativa. Os resultados mostraram que o prazo para o retorno de investimento está em 3,8 anos e que se espera que haja uma economia de cerca de R$ 2.753.000,00 nos próximos 25 anos, o que torna a instalação do sistema bastante atrativa. Sendo assim, é evidente o benefício que a utilização das fontes alternativas pode trazer tanto para as organizações, quanto ao meio ambiente, beneficiando a população como um todo. O Brasil tem um enorme potencial quanto o assunto, que é energia solar e com certeza pode ser mais bem explorada por todos segmentos da indústria/comércio brasileiro e mundial.

Made available in DSpace on 2021-06-25T11:02:27Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-06-01 Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) Pythiosis, whose etiological agent is the oomycete Pythium insidiosum, is a life-threatening disease that occurs mainly in tropical and subtropical countries, affecting several animal species. It is frequently found in horses in Brazil and humans in Thailand. The disease is difficult to diagnose because the pathogen’s hyphae are often misdiagnosed as mucoromycete fungi in histological sections. Additionally, there is no specific antigen to use for rapid diagnosis, the availability of which could improve the prognosis in different animal species. In this scenario, we investigated which P. insidiosum antigens are recognized by circulating antibodies in horses and humans with pythiosis from Brazil and Thailand, respectively, using 2D immunoblotting followed by mass spectrometry for the identification of antigens. We identified 23 protein spots, 14 recognized by pooled serum from horses and humans. Seven antigens were commonly recognized by both species, such as the heat-shock cognate 70 KDa protein, the heat-shock 70 KDa protein, glucan 1,3-beta-glucosidase, fructose-bisphosphate aldolase, serine/threonine-protein phosphatase, aconitate hydratase, and 14-3-3 protein epsilon. These results demonstrate that there are common antigens recognized by the immune responses of horses and humans, and these antigens may be studied as biomarkers for improving diagnosis and treatment. Department of Chemical and Biological Sciences Institute of Biosciences São Paulo State University (UNESP) Department of Pathology Faculty of Medicine Ramathibodi Hospital Mahidol University Faculty of Veterinary Medicine and Animal Science (FMVZ) São Paulo State University (UNESP) Department of Biological Sciences Bauru School of Dentistry University of São Paulo (USP) Department of Microbiology Immunology and Parasitology Cell Biology Division Federal University of São Paulo (UNIFESP) Center for the Study of Venoms and Venomous Animals (CEVAP) São Paulo State University (UNESP) Graduate Program in Tropical Diseases Botucatu Medical School (FMB) São Paulo State University (UNESP) Department of Chemical and Biological Sciences Institute of Biosciences São Paulo State University (UNESP) Faculty of Veterinary Medicine and Animal Science (FMVZ) São Paulo State University (UNESP) Center for the Study of Venoms and Venomous Animals (CEVAP) São Paulo State University (UNESP) Graduate Program in Tropical Diseases Botucatu Medical School (FMB) São Paulo State University (UNESP) CAPES: 001 FAPESP: 2016/10804-8 FAPESP: 2018/08009-0 FAPESP: 2019/08761-7

BACKGROUND PCM is a neglected systemic mycosis endemic in Brazil. The middle-west region of Brazil has shown the highest number of PCM by Paracoccidioides lutzii (P lutzii) cases. Differentiating cases of severe PCM from non-severe ones should be a concern at the bedside. Diagnosis of severe PCM by P lutzii is based on the subjectivity of clinical manifestations, which can result in a delay in starting its treatment and, consequently evolution to severe sequelae. There is not laboratory biomarker available to support the early diagnosis of severe PCM that is feasible for all the realities that coexist in Brazil. OBJECTIVES The aim of this study was to investigate the usefulness of laboratory biomarkers as erythrocyte sedimentation rate (ESR), C-reactive protein (CRP) and neutrophil/lymphocyte ratio (NLR) in the diagnosis of severe PCM. PATIENTS/METHODS ESR, CRP and NLR were analysed for 44 patients with PCM by P lutzii and a Receiver Operation Characteristic (ROC) curve were generated to identify the NLR cut-off point and point out the presence of severe PCM. RESULTS Sixteen (36.4%) had severe PCM and 28 (63.6%) had non-severe PCM. The mean NLR was higher and statistically significant among patients with severe PCM than among those with non-severe PCM. The area under the ROC curve was 0.859 for the diagnosis of severe PCM. The cut-off point for NLR for the diagnosis of severe PCM was 3.318 (sensitivity of 100%, specificity of 77%). CONCLUSIONS According to results, it is plausible to conclude that NLR represents a potential biomarker for the diagnosis of severe PCM.

This article shows reports of the clinical–epidemiological characteristics and serological screening in patients assisted by a reference center for PCM care, University Hospital Cassiano Antonio Moraes, Federal University of Espirito Santo, Brazil. The patient’s sera with PCM were analyzed by DID test at the beginning and the end treatment. Clinical and demographic data were also collected to characterize the sample. One hundred patients with a suspected diagnosis of PCM were evaluated. Serology by DID test was used as a screen in all patients. The test was positive for 79 patients (72 for Paracoccidioides brasiliensis and 7 for Paracoccidioides lutzii). Serology was negative in 21 sera, although all of them were diagnosed PCM by histopathologic or direct exam. Serological follow-up was performed during the treatment of all patients. After treatment, 58(58%) had negative serology and 33(33%) low levels of antibodies (≤ 1:16). Our results indicate the importance of the DID test for the screening and monitoring of PCM and that the incidence of P. lutzii might be greater than expected in areas where it is not the predominant PCM species. Therefore, this article may contribute to improving the knowledge and clinical management about this disease.

Paracoccidioidomycosis (PCM) is a life-threatening systemic fungal infection caused by members of the Paracoccidioides brasiliensis complex and P. lutzii. Routine diagnoses of PCM down to the species level using classical mycological approaches are unspecific due to overlapping phenotypes. There is an urgent need for specific, sensitive, and cost-effective molecular tools to diagnose PCM. Variation among the exon-2 of the gp43 gene was exploited to design species-specific primer pairs to discriminate between members of the P. brasiliensis complex and P. lutzii in a duplex PCR assay. Primer-BLAST searches revealed highly species-specific primers, and no significant region of homology was found against DNA databases except for Paracoccidioides species. Primers PbraCx-F and PbraCx-R targeting P. brasiliensis DNA produced an amplicon of 308 bp, while primers Plu-F and Plu-R targeting P. lutzii DNA generated an amplicon of 142 bp. The lower limit of detection for our duplex PCR assay was 1 pg of gDNA. A panel of 62 Paracoccidioides revealed 100% specificity (AUC = 1.000, 95%CI 0.972–1.000, p &lt, 0.0001) without cross-reacting with other medically relevant fungi or human DNA. As a proof of concept, we demonstrated the accurate identification of the P. brasiliensis complex (n = 7) or P. lutzii (n = 6) from a broad range of formalin-fixed, paraffin-embedded (FFPE) tissues of PCM patient’s organs. In four cases, FFPE PCR results confirmed, for the first time, co-infection due to P. brasiliensis (S1) and P. lutzii in the same biopsy. Our duplex PCR assay is useful to detect and differentiate members of the P. brasiliensis complex and P. lutzii, providing clinical laboratories with an important tool to be applied routinely, especially in atypical cases such as those featuring negative serology and positive mycological examination of clinical specimens as well as for the investigation of putative co-infection cases. This will likely benefit thousands of infected patients every year in a wide area of the Americas.

Made available in DSpace on 2022-04-29T08:46:09Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 Paracoccidioidomycosis (PCM) is a systemic mycosis caused by a group of cryptic species embedded in the Paracoccidioides brasiliensis complex and Paracoccidioides lutzii. Four species were recently inferred to belong to the P. brasiliensis complex, but the high genetic diversity found in both human and environmental samples have suggested that the number of lineages may be higher. This study aimed to assess the 43-kilodalton glycoprotein genotypes (PbGP43) in paraffin-embedded samples from PCM patients to infer the phylogenetic lineages of the P. brasiliensis complex responsible for causing the infection. Formalin-fixed, paraffin-embedded (FFPE) tissue samples from patients with histopathological diagnosis of PCM were analyzed. DNAs were extracted and amplified for a region of the second exon of the PbGP43 gene. Products were sequenced and aligned with other PbGP43 sequences available. A haplotype network and the phylogenetic relationships among sequences were inferred. Amino acid substitutions were investigated regarding the potential to modify physicochemical properties in the proteins. Six phylogenetic lineages were identified as belonging to the P. brasiliensis complex. Two lineages did not group with any of the four recognized species of the complex, and, interestingly, one of them comprised only FFPE samples. A coinfection involving two lineages was found. Five parsimony-informative sites were identified and three of them showed radical non-synonymous substitutions with the potential to promote changes in the protein. This study expands the knowledge regarding the genetic diversity existing in the P. brasiliensis complex and shows the potential of FFPE samples in species identification and in detecting coinfections. Centro de Diagnóstico e Pesquisa em Biologia Molecular Dr Ivo Ricci Departamento de Patologia Escola Paulista de Medicina Universidade Federal de São Paulo Departamento de Genética e Evolução Universidade Federal de São Carlos Disciplina de Biologia Celular Departamento de Microbiologia Imunologia e Parasitologia Universidade Federal de São Paulo Faculdade de Medicina de Botucatu Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP) LADR GmbH MVZ Dr, Kramer & Kollegen, Lauenburger Straße 67 Departamento de Ciências Farmacêuticas Universidade Federal de São Paulo, Campus Diadema Faculdade de Medicina de Botucatu Universidade Estadual Paulista Júlio de Mesquita Filho (UNESP)

Made available in DSpace on 2022-04-29T08:45:31Z (GMT). No. of bitstreams: 0 Previous issue date: 2021-01-01 Sporotrichosis is a subcutaneous mycosis resulting from the traumatic implantation of pathogenic Sporothrix species. In Brazil, zoonotic transmission plays an important role in the epidemiology of the disease, involving especially cats. The objective of this study was to isolate Sporothrix spp. from cats with signs of sporotrichosis, determining the causative species, clinical and epidemiological aspects, and the in vitro susceptibility profile of the isolates against antifungal drugs. From September 2017 to February 2019, 245 samples of lesions were collected from symptomatic cats in São José do Rio Preto, Brazil. Identification of the isolates was performed by morphophysiological parameters and species-specific polymerase chain reaction. The susceptibility profile of the isolates was determined for five drugs (amphotericin B, itraconazole, ketoconazole, potassium iodide and terbinafine), using the broth microdilution method. Clinical and epidemiological aspects were analyzed based on data contained on investigation forms filled by the veterinarians at moment of collection. Sporothrix spp. were isolated in 189 (77.2%) of the samples. Phenotypic and molecular analyses revealed S. brasiliensis as the only causative agent. In vitro susceptibility testing showed lower MIC values for terbinafine (MIC = 0.03—2 μg/ml), ketoconazole (MIC = 0.03—2 μg/ml), and itraconazole (MIC = 0.03—4 μg/ml). Most of the animals were male (73.5%), adults (96.3%), stray (53.5%), and uncastrated (69.8%). Our results show the expansion of the S. brasiliensis epidemic to an area nearly 840 km apart from the epicenter of the long-lasting outbreak of cat-transmitted sporotrichosis in Rio de Janeiro. Institute of Biosciences Humanities and Exact Sciences São Paulo State University (UNESP) Department of Dermatological Infectious and Parasitic Diseases School of Medicine (FAMERP) São José Do Rio Preto Laboratory of Emerging Fungal Pathogens Department of Microbiology Immunology and Parasitology Paulista School of Medicine Federal University of São Paulo Institute of Biosciences Humanities and Exact Sciences São Paulo State University (UNESP)

Histoplasmosis is a serious infectious disease in humans caused by Histoplasma spp. (Onygenales), whose natural reservoirs are thought to be soil enriched with bird and bat guano. The true global burden of histoplasmosis is underestimated and frequently the pulmonary manifestations are misdiagnosed as tuberculosis. Molecular data on epidemiology of Histoplasma are still scarce, even though there is increasing recognition of histoplasmosis in recent years in areas distant from the traditional endemic regions in the Americas. We used multi-locus sequence data from protein coding loci (ADP-ribosylation factor, H antigen precursor, and delta-9 fatty acid desaturase), DNA barcoding (ITS1/2+5.8s), AFLP markers and mating type analysis to determine the genetic diversity, population structure and recognise the existence of different phylogenetic species among 436 isolates of Histoplasma obtained globally. Our study describes new phylogenetic species and the molecular characteristics of Histoplasma lineages causing outbreaks with a high number of severe outcomes in Northeast Brazil between 2011 and 2015. Genetic diversity levels provide evidence for recombination, common ancestry and clustering of Brazilian isolates at different geographic scales with the emergence of LAm C, a new genotype assigned to a separate population cluster in Northeast Brazil that exhibited low diversity indicative of isolation. The global survey revealed that the high genetic variability among Brazilian isolates along with the presence of divergent cryptic species and/or genotypes may support the hypothesis of Brazil being the center of dispersion of Histoplasma in South America, possibly with the contribution of migratory hosts such as birds and bats. Outside Brazil, the predominant species depends on the region. We confirm that histoplasmosis has significantly broadened its area of occurrence, an important feature of emerging pathogens. From a practical point of view, our data point to the emergence of histoplasmosis caused by a plethora of genotypes, and will enable epidemiological analysis focused on understanding the processes that lead to histoplasmosis. Further, the description of this diversity opens avenues for comparative genomic studies, which will allow progress toward a consensus taxonomy, improve understanding of the presence of hybrids in natural populations of medically relevant fungi, test reproductive barriers and to explore the significance of this variation.

Paracoccidioidomycosis (PCM) is a life-threatening systemic infection caused by the fungal pathogen Paracoccidioides brasiliensis and related species. Whole-genome sequencing and stage-specific proteomic analysis of Paracoccidioides offer the opportunity to profile humoral immune responses against P. lutzii and P. brasiliensis s. str. infection using innovative screening approaches. Here, an immunoproteomic approach was used to identify PCM-associated antigens that elicit immune responses by combining 2-D electrophoresis of P. lutzii and P. brasiliensis proteomes, immunological detection using a gold-standard serum, and mass spectrometry analysis. A total of 16 and 25 highly immunoreactive proteins were identified in P. lutzii and P. brasiliensis, respectively, and 29 were shown to be the novel antigens for Paracoccidioides species, including seven uncharacterized proteins. Among the panel of proteins identified, most are involved in metabolic pathways, carbon metabolism, and biosynthesis of secondary metabolites in both immunoproteomes. Remarkably, six isoforms of the surface-associated enolase in the range of 54 kDa were identified as the major antigens in human PCM due to P. lutzii. These novel immunoproteomes of Paracoccidioides will be employed to develop a sensitive and affordable point-of-care diagnostic assay and an effective vaccine to identify infected hosts and prevent infection and development of human PCM. These findings provide a unique opportunity for the refinement of diagnostic tools of this important neglected systemic mycosis, which is usually associated with poverty.

This study investigated the effect of the quorum sensing molecules (QSMs) farnesol, 2-phenylehtanol, tyrosol and tryptophol against planktonic cells, filamentation and biofilms of