Abstract Background: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is the etiological agent of the disease coronavirus 2019 (COVID-19) in humans. SARS-CoV-2 has been identified in cats with or without clinical signs. Case presentation: We describe the pathological and molecular findings in a six-month-old asymptomatic cat with SARS-CoV-2 infection from Brazil, belonging to a human family with COVID-19 cases. The pool of nasopharynx and oropharynx swabs at day zero tested positive by RT-qPCR for SARS-CoV-2. No amplification resulted from molecular testing performed on days 7 and 14. The cat was hit by a car and died 43 days after the molecular diagnosis. Immunohistochemistry at post-mortem examination demonstrated nucleocapsid protein in samples from the lungs, kidneys, nasal conchae, trachea, intestine, brain and spleen. Conclusion: The present study has highlighted the possibility that viral antigens can be detected by immunohistochemistry in multiple organs six weeks after infection, although the same tissues tested negative by RT-PCR.

This study examined the association between oral candidiasis in elderly users and nonusers of prosthesis and its predisposing factors. To this end, we performed a cross-sectional study where saliva samples from 48 patients were collected they used prosthesis and 43 patients (control group) who did not use. Among the 91 patients, Candida spp were isolated in 40 (83.3%) who used prosthesis and in 23 (53.5%) in the control group. A statistically significant association was determined between the two groups, the isolation of yeasts and dental prosthesis (p < 0.05, OR = 4.3). The most common etiological agent was Candida albicans (37 isolates), with 23 (62.2%) in the denture group and 14 (37.8%) (control group). Among patients who presented clinical manifestations of oral candidiasis (n = 24), 83.3% (n = 20) belonged to the group that wore dentures, while only 16.7% (n = 4) belonged to the control group. Elderly patients with diabetes had 4.4 times higher estimated risk of developing oral candidiasis when compared with individuals without this condition. There was no statistically significant association between being user prostheses and have diabetes with the onset of candidiasis. No statistically significant association was determined between xerostomia, use of prosthesis and oral candidiasis. The use of prosthetics and poor oral hygiene in elderly patients predisposes to the development of oral candidiasis.

Candidemia permance como a maior causa de morbidade e mortalidade em ambiente hospitalar. A epidemiologia de infecções por Candida vem se alterando, principalmente em relação ao número de episódios causados por espécies não-albicans. Este estudo teve como objetivo avaliar a frequência, em um período de quatro anos, de leveduras do gênero Candida isoladas de sangue de pacientes pediátricos internados em hospital público da cidade de São Paulo, Brasil. Neste período foram isoladas leveduras de sangue de 104 pacientes, e as espécies de Candida identificadas, por métodos fenotípicos e genotípicos, foram: C. albicans (39/104), C. tropicalis (25/104), C. parapsilosis (23/104), Pichia anomala (6/104), C. guilliermondii (5/104), C. krusei (3/104), C. glabrata (2/104) e C. pararugosa (1/104). Em todo período do estudo foi observada maior frequência de isolamento de C. não-albicans (63,55%) (p = 0,0286). Neste estudo verificou-se aumento das espécies não-albicans ao longo dos anos (principalmente em 2009 e 2010), assim, ressalta-se que correta identificação em nível de espécie é recomendável, para que isso acarrete diagnóstico rápido e tratamento eficaz.

Candidemia remains a major cause of morbidity and mortality in the health care environment. The epidemiology of Candida infection is changing, mainly in relation to the number of episodes caused by species C. non-albicans. The overall objective of this study was to evaluate the frequency of yeasts of the genus Candida, in a four-year period, isolated from blood of pediatric patients hospitalized in a public hospital of the city of São Paulo, Brazil. In this period, yeasts from blood of 104 patients were isolated and, the identified species of Candida by phenotypic and genotypic methods were: C. albicans (39/104), C. tropicalis (25/104), C. parapsilosis (23/104), Pichia anomala (6/104), C. guilliermondii (5/104), C. krusei (3/104), C. glabrata (2/104) and C. pararugosa (1/104). During the period of the study, a higher frequency of isolates of C. non-albicans (63.55%) (p = 0.0286) was verified. In this study we verified the increase of the non-albicans species throughout the years (mainly in 2009 and 2010). Thus, considering the peculiarities presented by Candida species, a correct identification of species is recommended to lead to a faster diagnosis and an efficient treatment.

RESUMOA criptococose é micose sistêmica potencialmente grave causada por duas espécies do gêneroCryptococcus que acometem tanto homens como animais:Cryptococcus neoformans e C. gattii. São infecções cosmopolitas e emergentes, resultantes da interação do hospedeiro - humano e animalversus meio ambiente. A proposta deste trabalho foi avaliar a ocorrência de C. neoformans em 122 amostras de excretas secas de pombos coletadas em 49 locais na cidade de Cuiabá, Estado do Mato Grosso, Brasil, incluindo: praças públicas (n = 5), igrejas (n = 4), instituições de ensino (n = 3), unidades de saúde (n = 8), áreas abertas exibindo cobertura de amianto (n = 4), conjuntos residenciais domiciliares (n = 23), uma fábrica (n = 1) e um presídio (n = 1). Semeadura de suspensão de amostras em meio ágar niger (NSA), identificação fenotípica por provas bioquímicas e teste em meio de canavanina-glicina-azul de bromotimol, das colônias isoladas com pigmentação marrom escura. Foi também utilizada a técnica da reação em cadeia da polimerase com pares de iniciadores específicos para identificação de C. neoformans. As amostras foram coletadas de julho a dezembro de 2010. Cryptococcus neoformans foi isolado em oito (6,6%) de 122 amostras correspondendo a seis (12,2%) dos 49 sítios analisados.Cryptococcus neoformans associado a excretas de pombos ocorre em áreas de Cuiabá, predominando em residências nas amostras analisadas, constituindo fator de risco potencial para aquisição da doença tanto para indivíduos imunocomprometidos como imunocompetentes.

SUMMARYCryptococcosis is a severe systemic mycosis caused by two species of Cryptococcus that affect humans and animals: C. neoformans and C. gattii. Cosmopolitan and emergent, the mycosis results from the interaction between a susceptible host and the environment. The occurrence of C. neoformanswas evaluated in 122 samples of dried pigeon excreta collected in 49 locations in the City of Cuiabá, State of Mato Grosso, Brazil, including public squares (n = 5), churches (n = 4), educational institutions (n = 3), health units (n = 8), open areas covered with asbestos (n = 4), residences (n = 23), factory (n = 1) and a prison (n = 1). Samples collected from July to December of 2010 were seeded on Niger seed agar (NSA). Dark brown colonies were identified by urease test, carbon source assimilation tests and canavanine-glycine-bromothymol blue medium. Polymerase chain reaction primer pairs specific for C. neoformans were also used for identification. Cryptococcus neoformans associated to pigeon excreta was isolated from eight (6.6%) samples corresponding to six (12.2%) locations.Cryptococcus neoformans was isolated from urban areas, predominantly in residences, constituting a risk of acquiring the disease by immunocompromised and immunocompetent individuals.

INTRODUÇÃO: Contribuição sobre o perfil epidemiológico regional referente aos agentes fúngicos mais freqüentes nos Serviços de Saúde Pública em Cuiabá-MT, incluindo policlínicas e hospitais universitários. MÉTODOS: Foram examinados 1.496 espécimes clínicos colhidos a partir de 1.078 pacientes, os quais foram submetidos ao exame direto (potassa e/ou fita gomada) e cultivos em meios específicos. Os agentes foram identificados segundo micromorfologia (técnica de Ridell). RESULTADOS: Os 1.496 espécimes foram relacionados na maioria a exames de pele (n = 985), e unhas (n = 472). Dos 800 cultivos positivos, 246 (30,8%) corresponderam a dermatófitos, 336 (42%) a leveduras do gênero Candida, 190 (23,7%) a outras leveduras, 27 (3,4%) a fungos filamentosos não dermatofíticos e um (0,1%) a agente de micoses subcutâneas. Lesões consideradas primárias compareceram em maior número (59,5%), comparadas as recidivantes (37,4%). Foi observada maior positividade em membros inferiores e superiores. CONCLUSÕES: Co-morbidades, quadros alérgicos e diabetes mellitus representaram condições associadas à maior positividade em exames micológicos diretos e cultivos. O cultivo positivo foi considerado como diagnóstico definitivo de infecção fúngica, e confirmou 47,8% de hipóteses diagnósticas.

INTRODUCTION: A contribution to the regional epidemiological profile of the most common fungal agents in Public Health Services in Cuiabá, state of Mato Grosso, including university hospitals and polyclinics. METHODS: Clinical specimens (n = 1,496) from 1,078 patients were collected, submitted to direct mycological exam (potash or stick tape method) and cultured in specific mediums. Dermatophytic and non-dermatophytic agents were identified according to micromorphology (Ridell technique). RESULTS: The majority of the 1,496 specimens were skin (n = 985) and nail exams (n = 472). Of the 800 positive cultures, 246 (30.8%) corresponded to dermatophytes and 336 (42%) to yeasts of the genus Candida, 190 (23.7%) to other yeasts, 27 (3.4%) to non-dermatophytic filamentous fungi and one (0.1%) the agent of subcutaneous mycosis. Lesions considered primary occurred in greater numbers (59.5%) than recurrent lesions (37.4%), with a greater concentration of positivity occurring on the arms and legs. CONCLUSIONS: Comorbidities, allergies and diabetes mellitus were conditions associated with greater positivity in direct mycological exams and cultures. Positive culture was considered a definitive diagnosis of fungal infection and confirmed 47.8% of diagnostic hypotheses.

Causative agent in majority of VVC is Candida albicans, but infection due to non-C. albicans is common. Use of empiric antifungal therapy in Brazil due to syndromic management of vulvovaginitis could act as risk factor for increase resistance among VVC causative agents. From Mato Grosso patients, 160 with culture-proved among 404 women who had clinical symptoms of VVC, were enrolled in this study. 70 non-pregnant women and 90 pregnant women were included. Candida albicans was the most prevalent, representing 72.9% in the non-pregnant group and 92.3% in the pregnant group. Differences in species distribution were noted between the two groups, being C. parapsilosis the second more prevalent species among non-pregnant women. Susceptibility testing revealed high susceptibility to fluconazole (except for C. krusei), itraconazole, ketoconazole, and amphotericin B regardless the species (C. albicans, C. parapsilosis, C. tropicalis, C. glabrata, C. krusei) analyzed.

FUNDAMENTOS: Alguns fungos são habitantes do organismo humano e podem vir a causar alguma doença, quando há condições propícias para seu desenvolvimento. Infecções por leveduras são comuns e frequentes em pele e mucosas; contudo, espécies emergentes têm alterado o perfil epidemiológico. A habilidade de colonizar diversos sítios anatômicos tem sido associada à patogenicidade do gênero Candida, quando as condições ambientais são particularmente favoráveis. No caso de climas quentes e úmidos, os atritos sofridos pela pele ou as defesas imunitárias debilitadas podem fazer com que as leveduras deixem de ser comensais para se tornarem organismos patógenos. OBJETIVO: Diagnosticar candidíases em profissionais militares e avaliar a frequência dessas infecções nesses indivíduos. MÉTODOS: Os materiais clínicos analisados foram semeados em duplicata nos meios Sabouraud Dextrose-ágar (Difco) e Mycosel (Difco). Identificaram-se os agentes etiológicos por meio da observação de tubo germinativo, microcultivo e caracteres fisiológicos, assimilação de fontes de carbono (auxanograma) e fermentação de fontes de carbono (zimograma). RESULTADOS: De um total de 197 pacientes estudados, 91 (46,2%) apresentaram quadros clínicos de candidíases. A região genitocrural foi considerada a mais acometida (47,7%), seguida pelas regiões interdactilares (mãos e pés, 27,8%). C. albicans, C. parapsilosis, C. tropicalis, C. glabrata e espécies emergentes, como C. krusei e C. guilliermondii, foram identificadas. CONCLUSÃO: No ambiente de trabalho, o uso de calçados e de uniformes por longos períodos de tempo, associado ao estresse e à sudorese, foi considerado fator predisponente para o desenvolvimento das infecções fúngicas.

BACKGROUNDS: Some fungi are natural inhabitants of the human body but may result in disease when conditions are conducive to their development. Yeast infections are common and often occur in the skin and mucous membranes; however emerging species have changed this epidemiological profile. The ability to colonize different anatomical sites has been associated with the pathogenicity of Candida when environmental conditions are particularly favorable. In the case of hot, humid climates, the attrition suffered by the skin and weakened immune defenses may result in yeasts becoming pathogenic rather than commensal organisms. OBJECTIVE: The objective of this study was to diagnose yeast infections in military personnel and to evaluate the frequency of these infections in the individuals evaluated. METHODS: The clinical material analyzed was seeded in duplicate in Sabouraud dextrose agar (Difco™) and Mycosel medium (Difco™). The etiological agents were identified by observing the germ tubes, microculture and physiological characteristics, assimilation of carbon sources (auxanogram) and fermentation of carbon sources (zymogram). RESULTS: Of a total of 197 patients evaluated, 91 (46.2%) had episodes of candidiasis. The genitocrural region was the most commonly affected area (47.7%) followed by the interdigital regions (between the toes or fingers) (27.8%). Candida albicans, Candida parapsilosis, Candida tropicalis, Candida glabrata and emergent species such as Candida krusei and Candida guilliermondii were found. CONCLUSIONS: In the work environment, having to use shoes and uniforms for extended periods of time, in addition to stress and perspiration, were considered predisposing factors for the development of fungal infections.

INTRODUÇÃO: A candidíase é uma das infecções fúngicas mais frequentes entre os pacientes infectados pelo vírus da imunodeficiência humana. O presente estudo objetivou a caracterização das leveduras do gênero Candida de distintas amostras clínicas, provenientes de pacientes HIV - positivos, assim como a determinação do perfil de suscetibilidade in vitro a cinco drogas antifúngicas. MÉTODOS: A caracterização dos isolados de Candida sp foi realizada através da metodologia clássica, testes bioquímicos (zimograma e auxanograma) e morfológicos (prova do tubo germinativo e microcultivo em lâmina). Também, foram realizadas a técnica genotípica (PCR) e identificação pelo método comercial API 20C AUX (BioMeriéux). Para a determinação do perfil de suscetibilidade in vitro, foram utilizadas cinco drogas antifúngicas (cetoconazol, fluconazol, itraconazol, voriconazol e anfotericina B), através do método comercialmente disponível - Etest. RESULTADOS: Foram identificados 105 isolados de leveduras do gênero Candida provenientes de 102 pacientes infectados pelo vírus HIV. Destes, foram caracterizadas 82 (78,1%) Candida albicans, 8 (7,6%) Candida parapsilosis, 8 (7,6%) Candida tropicalis, 4 (3,8%) Candida krusei, 2 (1,9%) Candida glabrata e 1 (1%) Candida guilliermondii. CONCLUSÕES: Considerando o perfil geral de sensibilidade, 60% dos isolados foram suscetíveis a todos os antifúngicos testados, porém as espécies C. tropicalis e C. krusei demonstraram uma tendência a valores mais elevados de CIMs para os azóis do que os encontrados paraC. albicans, sugerindo resistência.

INTRODUCTION: Candidiasis is one of the most common fungal infections among patients infected by human immunodeficiency virus. The present study aimed to characterize yeasts of the genus Candida from distinct clinical samples from HIV-positive patients and determine the in vitro susceptibility profile to five antifungal drugs. METHODS: Characterization of Candida sp was achieved using the classic methodology: biochemical (zymogram and auxanogram) and micromorphology (germinative tube growth test and slide microculture) tests. Genotypic technique (PCR) and identification by the commercial method API 20C AUX (Biomeriéux) were also performed. To determine the in vitro susceptibility profile, five antifungal drugs were used (ketoconazole, fluconazole, itraconazole, voriconazole and amphotericin-B) following a commercially available method, the Etest. RESULTS: The procedure isolated 105 yeasts of the genus Candida from 102 HIV-infected patients. Of these, 82 (78.1%) were characterized as Candida albicans, 8 (7.6%) as C. parapsilosi s, 8 (7.6%) C. tropicalis, 4 (3.8%) C. krusei, 2 (1.9%) C. glabrata, and 1 (1%) as C. guiilliermondii. CONCLUSIONS: Considering the general profile of sensitivity, 60% of isolates were susceptible to all the antifungal drugs tested; however, the species C. tropicalis and C. krusei showed a tendency toward higher MICs to azoles than those obtained for C. albicans, suggesting resistance.

This study is the first report on genetic differences between isolates of Paracoccidioides brasiliensis from a single patient. We describe a simultaneous infection with genetically distinct isolates of P. brasiliensis in a patient with chronic paracoccidioidomycosis. The clinical isolates were obtained from lesions in different anatomical sites and were characterised by random amplified polymorphic DNA (RAPD) analysis. The RAPD technique can be helpful for distinguishing between clinical isolates. Different random primers were used to characterise these clinical isolates. The RAPD patterns allowed for differentiation between isolates and the construction of a phenetic tree, which showed more than 28% genetic variability in this fungal species, opening new possibilities for clinical studies of P. brasiliensis. Based on these results and preliminary clinical findings, we suggest that different genotypes of P. brasiliensis might infect the same patient, inducing the active form of the disease.

A prevalência de micose sistêmica entre 1.300 pacientes portadores de HIV/Aids de Cuiabá, Mato Grosso foi de 4,6%, no período de 2005-2008. As espécies de fungos isoladas foram o Cryptococcus neoformans (50%), Cryptococcus gattii (1,6%), Cryptococcus spp (6,6%), Histoplasma capsulatum (38,3%) e Paracoccidioides brasiliensis (3,3%). Óbito foi registrado em 32 (53,3%) pacientes, sendo a criptococose a principal causa. A contagem de linfócitos T CD4+ foi baixa e semelhante entre os pacientes que sobreviveram ou faleceram por micose sistêmica. O etilismo (OR:8,2; IC95%: 1,4-62,1; p=0,005) e o nível médio de desidrogenase lática [758 (182) U/L vs 416 (268) U/L; p<0,001] foram as características independentemente associadas ao óbito dos pacientes do estudo. Os resultados mostram alta letalidade por micoses sistêmicas em pacientes portadores de HIV/Aids de Cuiabá e sugerem que características clínico-laboratoriais tais como o etilismo e a elevação precoce da desidrogenase lática podem ser fatores relacionados ao pior prognóstico nessas condições.

Between 2005 and 2008, the prevalence of systemic mycosis among 1,300 HIV/AIDS patients in Cuiabá, Mato Grosso, was 4.6%. The fungus species isolated were Cryptococcus neoformans in 50%, Cryptococcus gattii in 1.6%, Cryptococcus spp in 6.6%, Histoplasma capsulatum in 38.3% and Paracoccidioides brasiliensis in 3.3%. Death was recorded in the cases of 32 patients (53.3%), and cryptococcosis was the main cause. The CD4+ T lymphocyte count was low and similar among patients who survived or died due to systemic mycosis. The factors independently associated with the deaths of these patients were alcoholism (OR: 8.2; 95% CI: 1.4-62.1; p = 0005) and the mean level of lactate dehydrogenase [758 (182) U/l vs. 416 (268) U/l; p < 0001]. The findings showed that systemic mycosis was highly lethal among the patients with HIV/AIDS in Cuiabá and suggested that clinical-laboratory characteristics such as alcoholism and early elevation of lactate dehydrogenase may be factors relating to worse prognosis under these conditions.

Foram avaliados 37 isolados de 10 pacientes HIV negativos e 26 positivos, em Mato Grosso. Exame direto, cultura e quimiotipagem de espécies foram realizados. Cetoconazol, itraconazol, voriconazol, fluconazol e anfotericina B foram avaliados. Foram identificadas 37 leveduras do gênero Cryptococcus spp sendo 26 de pacientes HIV- positivos (25 Cryptococcus neoformans e um Cryptococcus gattii) e 10 de HIV- negativos (cinco Cryptococcus neoformans e cinco Cryptococcus gattii). Considerando isolados clínicos (Cryptococcus neoformans) de HIV positivos observou-se resistência (8% e 8,7%) e susceptibilidade dose-dependência (20% e 17,4%) para fluconazol e itraconazol respectivamente. Para isolados de Cryptococcus neoformans oriundos de pacientes HIV negativos, observou-se susceptibilidade dose-dependência (40%) ao fluconazol. Os isolados de Cryptococcus gattii provenientes de pacientes HIV- negativos mostraram-se susceptíveis a todos os antifúngicos, exceto um isolado de Cryptococcus gattii que foi susceptível dose-dependente ao fluconazol (20%). O isolado proveniente do paciente HIV- positivo demonstrou resistência ao fluconazol (CIM > 256µg/mL) e itraconazol (CIM=3µg/mL).

Thirty-seven isolates from 10 HIV-negative and 26 HIV-positive patients in Mato Grosso were evaluated. Direct examination, culturing and chemotyping of species were performed. Ketoconazole, itraconazole, voriconazole, fluconazole and amphotericin B were evaluated. Thirty-seven yeasts of Cryptococcus spp were identified, of which 26 were from HIV-positive patients (25 Cryptococcus neoformans and one Cryptococcus gattii) and 10 from HIV-negative patients (five Cryptococcus neoformans and five Cryptococcus gattii). The Cryptococcus neoformans clinical isolates from HIV-positive patients showed resistance (8% and 8.7%) and dose-dependent susceptibility (20% and 17.4%) to fluconazole and itraconazole, respectively. Among the Cryptococcus neoformans isolates from HIV-negative patients, there was dose-dependent susceptibility (40%) to fluconazole. Cryptococcus gattii isolates from HIV-negative patients were shown to be susceptible to all antifungal agents, except for one isolate of Cryptococcus gattii that showed dose-dependent susceptibility to fluconazole (20%). The Cryptococcus gattii isolate from an HIV-positive patient showed resistance to fluconazole (MIC > 256 ¼g/ml) and itraconazole (MIC = 3 ¼g/ml).

Clinical cases involving paracoccidioidomycosis in children, diagnosed in Mato Grosso State, in the central western region of Brazil, are rare despite the state being classified with a moderate to high incidence. We describe a clinical case of infant acute disseminated Paracoccidioidomycosis in Mato Grosso State, Brazil, highlighting the importance of early differential diagnosis from other severe pathologies, thus contributing to the survival of compromised patients. A 3 year-old male child, weighing 12.8 Kg, originating from Vila Rica, MT, Brazil. The patient presented intermittent 40ºC fever evolving over 40 days, dry cough and painless bilateral cervical adenomegaly, showing no signs of inflammation. This was associated with diarrhea, distension and important abdominal pain and weight loss. Diagnosis was achieved by visualization of Paracoccidioides brasiliensis yeasts in a direct mycological exam and posterior fungus isolation in culture medium. The patient evolved presenting good clinical response to antifungal treatment and progressive reduction of abdominal and cervical ganglions. To improve the prognosis of compromised patients it is essential that professionals realize a full clinical-laboratorial evaluation, including differential diagnoses for other severe pathologies, as early as possible. The degree and intensity of paracoccidioidomycosis compromise are determining factors for defining the most efficient treatment.