CROSSOVER CLINICAL TRIAL OF THE INFLUENCE OF THE USE OF ADHESIVE ON BIOFILM FORMATION

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Antifungal Activity of Decyl Gallate against Several Species of Pathogenic Fungi

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Syngonanthus nitens (Bong.) Ruhland: caracterização biológica e prospecção terapêutica do extrato metanólico incorporado ou não em sistema nanoestruturado para aplicação no tratamento da candidíase vulvovaginal

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Could histoplasma capsulatum be related to healthcare-associated infections?

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Nasal, oral and rectal microbiota of Black lion tamarins (Leontopithecus chrysopygus)

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Microbiota fúngica de passeriformes de cativeiros da Região Noroeste do Estado de São Paulo

Birds are hosts for a rich fungal microbiota which can act as potent pathogens for humans and other species of animals, causing thereby serious public health problems. The objective of this study was to evaluate the participation of birds kept in containers in the epidemiology of infectious diseases such as cryptococcosis and aspergillosis, thus verifying the maintenance and spread of pathogens in the environment. 36 samples of excretas of passeriformes were collected and were cultivated in Sabouraud Dextrose Agar 4% at room temperature and 37°C. The isolated fungal colonies were classified according to their morphological and staining characteristics. Subsequently, those in yeast form were peaked in Niger Agar, incubated at 30°C. In one sample showed growth of more than one type of colony and there was verified the presence of 25.0% of Penicillium spp., 19.4% of Trichosporon spp., 13.9% of C. gattii, 11.1% of C. neoformans, 11.1% of Candida spp., 8.3% of Rhizomucor spp., 8.3% of Aspergillus spp., 2.8% of Nigrospora spp. and 2,8% of Geotrichum spp. It can be conluded by the expost that birds shed continuously pathogenic microorganisms in their feces acting in definitive form in the infectious diseases ecoepidemiology.

Antimicrobial Photodynamic Therapy in the Treatment of Oral Candidiasis in HIV-Infected Patients

Objective: Antimicrobial photodynamic therapy (aPDT) has been used to combat local infections, and it consists of the combination of a photosensitizer, a light source, and reactive oxygen species (ROS) to kill microbial cells. In this study, we evaluated the effectiveness of aPDT in the treatment of candidiasis in HIV-infected patients. Methods: Twenty-one patients were divided into three groups. Control group (CG) was treated with the conventional medication for candidiasis (fluconazole 100 mg/day during 14 days). Laser group (LG) was subjected to low-level laser therapy (LLLT), wavelength 660 nm, power of 30 mW, and fluence of 7.5 J/cm(2), in contact with mucosa during 10 sec on the affected point. An aPDT group (aPDTG) was treated with aPDT, that is, combination of a low-power laser and methylene blue 450 mu g/mL. Pre-irradiation time was 1 min. Parameters of irradiation were the same ones as for the LG, and patients were single irradiated. Patients were clinically evaluated and culture analysis was performed before, immediately after, and 7, 15, and 30 days after the treatment. Results: Our results showed that fluconazole was effective; however, it did not prevent the return of the candidiasis in short-term. LLLT per se did not show any reduction on Candida spp. aPDT eradicated 100% of the colonies of this fungus and the patients did not show recurrence of candidiasis up to 30 days after the irradiation. Conclusions: These findings suggest that aPDT is a potential approach to oral candidiasis treatment in HIV-infected patients.

Effects of eating disorders on oral fungal diversity

Background. The eating disorders anorexia and bulimia nervosa can cause several systemic and oral alterations related to poor nutrition and induced vomiting; however, the oral microflora of these patients is poorly studied. Objective. The aim of this study was to evaluate fungal microflora in the oral cavity of these patients by culture-dependent and culture-independent methods. Study Design. Oral rinse samples were cultured to assess the prevalence of Candida species, and the isolates were identified by API system. Microorganism counts were compared by the Mann-Whitney test (5%). Ribotyping, a type of molecular analysis, was performed by sequencing the D1/D2 regions of 28S rRNA. Results. Our results demonstrated that the eating disorder group showed higher oral Candida spp. prevalence with culture-dependent methods and higher species diversity with culture-independent methods. Conclusions. Eating disorders can lead to an increased oral Candida carriage. Culture-independent identification found greater fungal diversity than culture-dependent methods. (Oral Surg Oral Med Oral Pathol Oral Radiol 2012;113:512-517)

Epidemiological and clinical characteristics of nosocomial candidiasis in university hospitals in Cuiaba - Mato Grosso, Brazil

Background: Fungal infections are emerging as an important cause of human disease, especially among hospitalized patients with serious underlying disease and several risk factors. Aims: To evaluate epidemiological and clinical characteristics of patients with nosocomial candidiasis in university hospitals in Cuiaba - MT, Brazil. Methods: A descriptive study of 91 patients admitted to university hospitals in Cuiaba - MT, with clinical and laboratory diagnosis of nosocomial candidiasis, over a 20-month period. Results: A rate for nosocomial infections by Candida spp. of 5 per 1000 admissions, proportional mortality of 14.4% and lethality of 53.8% were determined. The patient age ranged from 29 days to 82 years-old, among which, 74.7% were adults and 25.3% children. The intensive care units contributed with the highest number of cases of infection by Candida spp. (69.2%). The most important underlying disease was gastrointestinal tract disease (11%). Prematurity and low birth weight were the most important risk factors among newborns. The use of antibiotics, invasive procedures, H-2 blockers, multiple blood transfusions and stay length of >= 21 days were the most frequent risk factors among adults. Candida albicans was the most common species in all cases. Conclusions: In this study, C. albicans was the most frequently detected species in candidiasis and risk factors increased the susceptibility of hospitalized patients to acquiring a nosocomial infection by Candida spp. (C) 2011 Revista Iberoamericana de Micologia. Published by Elsevier Espana, S.L. All rights reserved.

Photodynamic inactivation of microorganisms present on complete dentures. A clinical investigation

The aim of this study was to evaluate the effectiveness of photodynamic therapy (PDT) for the disinfection of complete dentures. Biofilm samples were collected from dentures of 60 denture users who were randomly divided into four experimental groups (n = 15 each): subjects whose maxillary dentures were sprayed with 50 and 100 mg/l of PhotogemA (R) suspension (groups P50S and P100S) and patients whose maxillary dentures were treated with 50 and 100 mg/l of PhotogemA (R) gel (groups P50G and P100G). Dentures with photosensitizers were left in the dark for 30 min (pre-irradiation time) and then irradiated with blue LED light at 37.5 J/cm(2) (26 min). Denture samples were taken with sterile cotton swab before (left side surfaces) and after (right side surfaces) PDT. All microbial material was diluted and plated on selective media for Candida spp., Staphylococcus mutans spp., streptococci and a non-selective media. After incubation (48 h/37A degrees C), the number of colony-forming units (cfu/ml) was counted. Microorganisms grown on selective media were identified using biochemical methods before and after PDT. The data were submitted to McNemar and Kruskal-Wallis tests (alpha = 0.05). No growth after PDT was observed in 60, 53, 47, and 40% of dentures from P100G, P50G, P100S, and P50S groups, respectively. When evidence of microorganisms' growth was observed, PDT regimens eliminated over 90% of microorganisms on dentures. This clinical study showed that PDT was effective for disinfecting dentures.

Invasive candidiasis and oral manifestations in premature newborns

Objective: To investigate prevalence of invasive candidiasis in a Neonatal Intensive Care Unit and to evaluate oral diseases and Candida spp. colonization in low birth weight preterm newborns. Methods: A descriptive epidemiological study performed in two stages. First, prevalence of candidiasis was analyzed in a database of 295 preterm patients admitted to hospital for over 10 days and birth weight less than 2,000g. In the second stage, oral changes and Candida spp. colonization were assessed in 65 patients weighing less than 2,000g, up to 4 week-old, hospitalized for over 10 days and presenting oral abnormalities compatible with fungal lesions. Swab samples were collected in the mouth to identify fungi. Results: Prevalence of candidiasis was 5.4% in the database analyzed. It correlated with prolonged hospital length of stay (p<0.001), in average, 31 days, and 85% risk of developing infection in the first 25 days. It correlated with low birth weight (p<0.001), with mean of 1,140g. The most frequent alterations were white soft plaques, detachable, in oral mucosa and tongue. Intense oral colonization by Candida spp was observed (80%). Conclusions: The frequency of invasive candidiasis was low and correlated with low birth weight and prolonged hospital stay. The most common oral changes were white plaques compatible with pseudomembranous candidiasis and colonization by Candida spp. was above average.

Diferença de tempo de positividade: método útil no diagnóstico de infecção de corrente sanguínea relacionada com cateter?

INTRODUÇÃO: As infecções de corrente sanguínea relacionadas com cateter (ICSRCs) apresentam impacto significativo na morbidade e na mortalidade de pacientes internados, além de elevar custos hospitalares. A utilização de equipamentos automatizados no processamento de hemoculturas gerou uma alternativa para diagnóstico de ICSRC por meio da análise da diferença de tempo de positividade (DTP) entre hemoculturas pareadas (coletadas simultaneamente) de sangue periférico e sangue de cateter. Um diagnóstico acurado e rápido dessas infecções pode otimizar as condutas clínicas e terapêuticas, poupando a retirada precoce dos cateteres. OBJETIVOS: Avaliar na rotina a DTP como ferramenta auxiliar no diagnóstico de ICSRC e determinar os principais microrganismos isolados. MÉTODOS: Foram avaliadas retrospectivamente hemoculturas coletadas no complexo do Hospital das Clínicas da Faculdade de Medicina da Universidade de São Paulo (HC/FMUSP) de maio a agosto de 2008. Somente amostras que apresentaram DTP maior que 120 minutos foram consideradas possíveis ICSRCs pelo critério laboratorial. RESULTADOS: A seção processou 11.017 hemoculturas aeróbias durante o período de estudo; somente 5% foram coletadas de forma pareada. Destas, 148 (28%) foram positivas, sendo 9% com crescimento somente em sangue periférico, 41% somente em sangue de cateter e 50% em ambas as amostras com 88% de homologia de microrganismos identificados. A DTP apresentou valores acima de 120 minutos em 50% dos casos e os microrganismos mais isolados foram Staphylococcus aureus (22%), Candida spp. (18%), Klebsiella spp. (7%) e Enterobacter spp. (7%). CONCLUSÃO: A determinação da DTP como ferramenta auxiliar no diagnóstico de ICSRC é viável e fácil de ser executada em laboratórios de rotina com automação, porém o processo de coleta das amostras pareadas deve ser rigidamente padronizado.

Carbon and nitrogen limitation increase chitosan antifungal activity in Neurospora crassa and fungal human pathogens

Chitosan permeabilizes plasma membrane and kills sensitive filamentous fungi and yeast. Membrane fluidity and cell energy determine chitosan sensitivity in fungi. A five-fold reduction of both glucose (main carbon (C) source) and nitrogen (N) increased 2-fold Neurospora crassa sensitivity to chitosan. We linked this increase with production of intracellular reactive oxygen species (ROS) and plasma membrane permeabilization. Releasing N. crassa from nutrient limitation reduced chitosan antifungal activity in spite of high ROS intracellular levels. With lactate instead of glucose, C and N limitation increased N. crassa sensitivity to chitosan further (4-fold) than what glucose did. Nutrient limitation also increased sensitivity of filamentous fungi and yeast human pathogens to chitosan. For Fusarium proliferatum, lowering 100-fold C and N content in the growth medium, increased 16-fold chitosan sensitivity. Similar results were found for Candida spp. (including fluconazole resistant strains) and Cryptococcus spp. Severe C and N limitation increased chitosan antifungal activity for all pathogens tested. Chitosan at 100 μg ml-1 was lethal for most fungal human pathogens tested but non-toxic to HEK293 and COS7 mammalian cell lines. Besides, chitosan increased 90% survival of Galleria mellonella larvae infected with C. albicans. These results are of paramount for developing chitosan as antifungal.

Omics for Investigating Chitosan as an Antifungal and Gene Modulator

Chitosan is a biopolymer with a wide range of applications. The use of chitosan in clinical medicine to control infections by fungal pathogens such as Candida spp. is one of its most promising applications in view of the reduced number of antifungals available. Chitosan increases intracellular oxidative stress, then permeabilizes the plasma membrane of sensitive filamentous fungus Neurospora crassa and yeast. Transcriptomics reveals plasma membrane homeostasis and oxidative metabolism genes as key players in the response of fungi to chitosan. A lipase and a monosaccharide transporter, both inner plasma membrane proteins, and a glutathione transferase are main chitosan targets in N. crassa. Biocontrol fungi such as Pochonia chlamydosporia have a low content of polyunsaturated free fatty acids in their plasma membranes and are resistant to chitosan. Genome sequencing of P. chlamydosporia reveals a wide gene machinery to degrade and assimilate chitosan. Chitosan increases P. chlamydosporia sporulation and enhances parasitism of plant parasitic nematodes by the fungus. Omics studies allow understanding the mode of action of chitosan and help its development as an antifungal and gene modulator.

Candidemia : revisão e atualização

Trabalho Final do Curso de Mestrado Integrado em Medicina, Faculdade de Medicina, Universidade de Lisboa, 2014