Rapid Molecular Diagnosis of Sporotrichosis Directly from Biological Samples from a Reference Center in Brazil.

The gold standard diagnosis of sporotrichosis is the isolation of Sporothrix sp. in culture media, but this is a time-consuming test that is susceptible to contamination and can be affected by the fungal load. Molecular methods such as nested PCR are gaining more ground in the management of several infections as they are tools for the rapid and accurate identification of microorganisms from pure cultures or directly from biological samples. This study aimed to apply a nested PCR molecular protocol for the rapid detection of Sporothrix spp. directly from clinical samples. Thirteen samples-six from skin biopsies, five from skin exudates, and two from conjunctival secretions-were obtained from patients diagnosed with sporotrichosis due to S. brasiliensis. Calmodulin gene sequencing identified all the isolates as S. brasiliensis. Nested PCR was able to detect all the Sporothrix sensu lato directly from clinical samples as well as the CBS 120339 reference strain. The nested PCR protocol stands out as a diagnostic alternative, as it allows the identification of Sporothrix spp. directly from clinical samples without the need for fungal isolation.

ß-Estradiol and progesterone enhance biofilm development and persister cell formation in monospecies and microcosms biofilms derived from vulvovaginal candidiasis.

The present study aimed to: (1) evaluate the influence of the steroid hormones (SH) on biofilm development; (2) investigate the formation of persister cells (PC) in biofilms; and (3) investigate the influence of SH on PC formation. Biofilms were derived from vulvovaginal candidiasis (VVC) samples and evaluated by three models: microcosm biofilms grown in Vaginal Fluid Simulator Medium (MiB-VFSM); monospecies biofilms grown in VFSM (MoB-VFSM) and RPMI media (MoB-RPMI). SH altered cell counting and biomass of biofilms grown in VSFM; MoB-RPMI were negatively affected by SH. SH stimulated the formation of PC in MiB-VFSM but not MoB-VFSM; MoB-RPMI showed a lower number of PC in the presence of SH. The results showed that SH altered the dynamics of biofilm formation and development, depending on the study model. The data suggest the influence of hormones on the physiology of Candida biofilms and reinforce the importance of PC in the pathogenesis of VVC.

Atypical chlamydoconidium-producing Trichophyton tonsurans strains from Ceará State, Northeast Brazil: investigation of taxonomy by phylogenetic analysis and biofilm susceptibility.

Chlamydoconidium-producing Trichophyton tonsurans strains isolated in Northeastern Brazil have morphological features different from the classic description of this dermatophyte species. This study investigated the phylogenetic relationship of chlamydoconidium-producing T. tonsurans strains isolated in Northeastern Brazil. Also, the effect of terbinafine and farnesol on mature biofilms of T. tonsurans strains was evaluated. The mass spectra of T. tonsurans strains were investigated by matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS). The ITS and LSU loci regions of rDNA and the partial ß-tubulin gene were sequenced and the phylogenetic tree was analysed. The effects of terbinafine and farnesol on mature T. tonsurans biofilms were evaluated through the analysis of metabolic activity, quantification of biomass and observation by scanning electron microscopy. MALDI-TOF MS spectra of the chlamydoconidium-producing T. tonsurans strains differed from the spectrum of the control strain (ATCC 28942), presenting an intense ion peak at m/z 4155 Da. Phylogenetic tree analysis showed that the chlamydoconidium-producing strains isolated in Northeastern Brazil are allocated to a single cluster, differing from strains isolated from other countries. As for mature T. tonsurans biofilms, farnesol reduced biomass and metabolic activity by 64.4 and 65.9 %, respectively, while terbinafine reduced the biomass by 66.5 % and the metabolic activity by 69 %. Atypical morphological characteristics presented by chlamydoconidium-producing T. tonsurans strains result from phenotypic plasticity, possibly for adaptation to environmental stressors. Also, farnesol had inhibitory activity against T. tonsurans biofilms, demonstrating this substance can be explored for development of promising anti-biofilm drugs against dermatophytes.

Is Sporothrix chilensis circulating outside Chile?

Sporothrix chilensis is a mild-pathogenical specie of Sporothrix pallida complex, until now, known as restrict to Chile. Herein, we describe the first clinical isolates identified as S. chilensis in Brazil, preserved in the URM Culture Collection, by polyphasic taxonomy, and their respective antifungal profile of this emergent fungus.

Candidemia in a public hospital in Northeastern Brazil: epidemiological features and risk factors in critically ill patients / Candidemia en un hospital público del noreste de Brasil: aspectos epidemiológicos y factores de riesgo en los pacientes críticos

Background: Candidemia is a life-threatening fungal infection characterized by the presence of Candida in the blood. Aims: To describe the clinical-epidemiological features and main risk factors among patients with candidemia admitted to Intensive Care Unit. Methods: A cross-sectional, retrospective and observational study was performed between January 2015 and July 2016. Laboratory reports and medical records from ICU patients admitted to a public hospital in northeastern Brazil were analyzed. Results: There were 1573 admissions and 67 of them were positive for candidemia. The majority of patients were male (53.3%) and remained at the hospital for more than seven days (86.6%). Non-C. albicans Candida infections (60%) were predominant. Broad-spectrum antibiotic therapy was prescribed in 98.4% of the cases. The most frequent underlying diseases were sepsis (73.3%), presence of solid tumors (15%), respiratory condition (60%), urinary tract disease (56.6%) and gastrointestinal tract diseases (23.3%). Surgeries were carried out on 43% of the patients, consisting of 23.3% abdominal surgeries, with a mortality rate of 92.8%. Risk factors were venous central access (93.3%), mechanical ventilation (81.6%), nasoenteral tube (83.3%), nasogastric tube (25%), indwelling bladder catheter (88.3%), diabetes mellitus (55%) and tracheostomy (36.6%). Statistical analysis correlated the use of indwelling bladder catheter with a higher mortality rate (r=0.07412, p=0.0353). Conclusions: The current study reveals the high case fatality rates among critically ill patients suffering from candidemia admitted to ICU. Herein, we highlight the importance of identifying non-C. albicans Candida species and reinforce the idea of carrying out epidemiological surveillances and antifungal susceptibility tests

Antecedentes: La candidemia es una infección potencialmente fatal caracterizada por la presencia de Candida en la sangre. Objetivos: Describir las características clínico-epidemiológicas y los principales factores de riesgo en pacientes con candidemia ingresados en la unidad de cuidados intensivos (UCI). Métodos: Entre enero de 2015 y julio de 2016 se llevó a cabo un estudio transversal, retrospectivo y observacional en el que se analizaron los registros médicos e informes de laboratorio de pacientes de la UCI de un hospital público del noreste de Brasil. Resultados: Entre las 1.573 admisiones registradas hubo 67 diagnósticos de candidemia. La mayoría de los pacientes fueron del sexo masculino (53,3%) y la permanencia en el hospital fue superior a siete días (86,6%). Las infecciones por especies de Candida no-C. albicans fueron el 60% de los casos. En el 98,4% de los casos se prescribió antibioterapia de amplio espectro. Las enfermedades de base más frecuentes fueron la sepsis (73,3%), la presencia de tumores sólidos (15%), las enfermedades respiratorias (60%), la enfermedad del tracto urinario (56,6%) y las enfermedades del tracto gastrointestinal (23,3%). Se realizaron cirugías en el 43% de los pacientes, siendo el 23,3% cirugías abdominales, con una tasa de mortalidad del 92,8%. Los factores de riesgo fueron la existencia de un acceso venoso central (93,3%), ventilación mecánica (81,6%), sonda nasoenteral (83,3%), sonda nasogástrica (25%), catéter vesical permanente (88,3%), diabetes mellitus (55%) y traqueostomía (36,6%). Los análisis estadísticos correlacionaron el uso del catéter urinario permanente con una mayor mortalidad (r=0,07412; p=0,0353). Conclusiones: Este estudio muestra las elevadas tasas de letalidad asociadas a pacientes con candidemia ingresados en la UCI. Destacamos la creciente importancia de identificar las especies de Candida diferentes de C. albicans, y reforzamos la idea de llevar a cabo una vigilancia epidemiológica y de realizar pruebas de sensibilidad a los antifúngicos

Coccidioidomycosis: first cases reported in Pernambuco, Brazil.

Coccidioidomycosis is a fungal infection caused by Coccidioides immitis or Coccidioides posadasii. These fungi are known to thrive in desert climate. Fungi produce infectious arthroconidia in soil, they are aerosolized in the air and when inhaled by humans, usually cause infections such as pneumonia. The first cases of coccidioidomycosis in Brazil were reported in 1978. Since then, there have been other reports mainly from desert regions of Northeastern Brazil. The present report describes three cases of coccidioidomycosis on male farmers from Serra Talhada county, Pernambuco State, who developed pneumonia and were subsequently diagnosed with pulmonary coccidioidomycosis. These three farmers were successfully treated with oral fluconazole. They reported having hunted armadillos in a rural and arid area of Pernambuco State. Armadillos are known to be carriers of Coccidioides. This is the first report of infection caused by Coccidioides in Pernambuco State, Brazil.

The HIV aspartyl protease inhibitor ritonavir impairs planktonic growth, biofilm formation and proteolytic activity in Trichosporon spp.

This study evaluated the effect of the protease inhibitor ritonavir (RIT) on Trichosporon asahii and Trichosporon inkin. Susceptibility to RIT was assessed by the broth microdilution assay and the effect of RIT on protease activity was evaluated using azoalbumin as substrate. RIT was tested for its anti-biofilm properties and RIT-treated biofilms were assessed regarding protease activity, ultrastructure and matrix composition. In addition, antifungal susceptibility, surface hydrophobicity and biofilm formation were evaluated after pre-incubation of planktonic cells with RIT for 15 days. RIT (200 µg ml-1) inhibited Trichosporon growth. RIT (100 µg ml-1) also reduced protease activity of planktonic and biofilm cells, decreased cell adhesion and biofilm formation, and altered the structure of the biofilm and the protein composition of the biofilm matrix. Pre-incubation with RIT (100 µg ml-1) increased the susceptibility to amphotericin B, and reduced surface hydrophobicity and cell adhesion. These results highlight the importance of proteases as promising therapeutic targets and reinforce the antifungal potential of protease inhibitors.

Cryptococcus laurentii fungaemia in a cervical cancer patient

ABSTRACT Infections caused by emerging Cryptococcus non-neoformans species are being reported with increasingly frequency. Here, we present a case of fungaemia byCryptococcus laurentii in a woman receiving aggressive immunosuppressive therapy for cervical neoplasia. Three venous blood samples were aseptically collected on consecutive days and C. laurentiiwas isolated and identified through phenotypic and molecular methods. After central venous catheter removal and appropriate antifungal therapy, the patient showed significant improvement and blood culture became negative. Thus, patients following immunosuppressive therapies and using invasive medical devices are at risk of C. laurentii blood infections.

Cryptococcus laurentii fungaemia in a cervical cancer patient.

Infections caused by emerging Cryptococcus non-neoformans species are being reported with increasingly frequency. Here, we present a case of fungaemia by Cryptococcus laurentii in a woman receiving aggressive immunosuppressive therapy for cervical neoplasia. Three venous blood samples were aseptically collected on consecutive days and C. laurentii was isolated and identified through phenotypic and molecular methods. After central venous catheter removal and appropriate antifungal therapy, the patient showed significant improvement and blood culture became negative. Thus, patients following immunosuppressive therapies and using invasive medical devices are at risk of C. laurentii blood infections.

Invasive infection due to Saprochaete capitata in a young patient with hematological malignancies

We report a case of invasive infection due to Saprochaete capitata in a patient with hematological malignancies after chemotherapy treatment and empiric antifungal therapy with caspofungin. Although severely immunocompromised the patient survived been treated with amphotericin B lipid complex associated with voriconazole.

[Opportunistic yeast infections and enzymatic profile of the etiological agents]. / Infecções oportunistas por leveduras e perfil enzimático dos agentes etiológicos.

Yeast infections are common in immunocompromised patients, although emerging species have been changing the epidemiological profile. The ability to secrete proteinases has been associated with pathogenicity within the genus Candida. This study had the aims of diagnosing yeast infections in immunocompromised patients and evaluating the virulence of the etiological agents, based on a proteinase secretion assay using bovine serum albumin as a substrate. Out of a total of 104 patients studied, 19 presented episodes of yeast infection. The respiratory tract (63.2%), followed by the urinary tract (10.5%), were the most common sites of infection. Candida albicans, Candida parapsilosis, Candida tropicalis and emerging species such as Candida krusei and Candida guilliermondii were isolated. Five isolates of Candida parapsilosis and one of Candida albicans and Candida guilliermondii exhibited high enzymatic activity. We conclude that enzymatic characterization of Candida isolates may be a useful prognostic marker, especially among immunocompromised individuals, since yeast infections in such patients are generally serious.

Infecções oportunistas por leveduras e perfil enzimático dos agentes etiológicos / Opportunistic yeast infections and enzymatic profile of the etiological agents

Infecções por leveduras são freqüentes em imunocomprometidos, contudo espécies emergentes têm alterado o perfil epidemiológico. A habilidade de secretar proteases tem sido associada à patogenicidade do gênero Candida. Esta pesquisa teve como objetivos diagnosticar leveduroses em pacientes imunocomprometidos e avaliar a virulência dos agentes etiológicos baseado em teste de secreção de protease utilizando soro de albumina bovina como substrato. Do total de 104 pacientes estudados, 19 apresentaram episódios de leveduroses. O trato respiratório (63,2 por cento), seguido pelo trato urinário (10,5 por cento) foram os locais mais comuns de infecção. Candida albicans, Candida parapsilosis, Candida tropicalis e espécies emergentes como Candida krusei e Candida guilliermondii foram isoladas. Cinco isolados de Candida parapsilosis e um de Candida albicans e Candida guilliermondii exibiram alta atividade enzimática. Concluímos que a caracterização enzimática de isolados de Candida pode ser um útil marcador prognóstico, especialmente em imunocomprometidos, uma vez que leveduroses nestes pacientes são geralmente graves.

Yeast infections are common in immunocompromised patients, although emerging species have been changing the epidemiological profile. The ability to secrete proteinases has been associated with pathogenicity within the genus Candida. This study had the aims of diagnosing yeast infections in immunocompromised patients and evaluating the virulence of the etiological agents, based on a proteinase secretion assay using bovine serum albumin as a substrate. Out of a total of 104 patients studied, 19 presented episodes of yeast infection. The respiratory tract (63.2 percent), followed by the urinary tract (10.5 percent), were the most common sites of infection. Candida albicans, Candida parapsilosis, Candida tropicalis and emerging species such as Candida krusei and Candida guilliermondii were isolated. Five isolates of Candida parapsilosis and one of Candida albicans and Candida guilliermondii exhibited high enzymatic activity. We conclude that enzymatic characterization of Candida isolates may be a useful prognostic marker, especially among immunocompromised individuals, since yeast infections in such patients are generally serious.