Epidemiologia de candidíase hospitalar: Importância da identificação específica

Nosocomial infections with Candida species are recognized as a significant cause of morbidity and mortality in both seriously ill immunocompetent and immunocompromised patients. Infections with Candida albicans and non-albicans Candida species have become a significant cause of infection in humans. Several of the more commonly Candida spp isolates are less susceptible to the antifungal drugs currentlly applied in clinical treatment, a factor that means significant difficulties for effective treatment. The modern mycology laboratory has an important role to play in several aspects relating to these organisms, including therapy, detection, identification and epidemiological analysis. In this study, we have provided an initial comparison of differences in species distribution among Candida isolates from four general hospitals of São Paulo,SP. Overall, 40 isolates of C. albicans, C. parapsilosis and C. tropicalis were isolated respectively in 35%, 50% and 15%, revealed a tendency of higher frequency of non-albicans species. The species distribution in patients with candidemia showed that the most commonly species were C. parapsilosis (45,4%), followed by C. albicans (36,4%) and C. tropicalis (18,2%); thus, we have an increase of non-albicans species. The three different species were include in 6, 3, and 4 different biotypes, respectively C. albicans, C. parapsilosis e C. tropicalis. This study emphasizes the importance of periodic evaluation of Candida species distribution especially in centers caring for patients at risk.

In vitro antifungal susceptibility of Candida spp. isolates from patients with chronic periodontitis and from control patients

Superinfection by Candida can be refractory to conventional periodontal treatments in specific situations, such as in immunocompromised patients. In these cases, the systemic therapy with antifungal drugs could be indicated. The aim of this study was to analyse antifungal susceptibility of Candida spp. strains isolated from chronic periodontitis patients and from control individuals. A total of 39 C. albicans isolates, 9 C. tropicalis, 2 C. glabrata and 5 Candida spp. from control individuals and 30 C. albicans, 3 C. tropicalis and 2 C. glabrata from periodontitis patients were tested. In the control group, 1 isolate of C. glabrata was resistant to ketoconazole and 1 Candida spp. was resistant to amphotericin B, ketoconazole and miconazole. Among the isolates of periodontitis group, 1 (3.33%) C. albicans isolate was resistant to flucytosine and ketoconazole. According to the obtained results, it could be concluded that fluconazole was the most effective drug against the several Candida species studied. There were not expressive differences in the susceptibility of isolates from periodontitis patients or from control individuals.

2,3,4,6-Tetra-O-(3-nitropropanoyl)-O-β-D-glucopyranoside, a new antimicrobial from the roots of Heteropteris aphrodisiaca

A new 2,3,4,6-tetra-O-(3-nitropropanoyl)-O-β-D-glucopyranoside anomer was isolated from the roots of Heteropteris aphrodisiaca and characterized by spectroscopic methods. Activity against Staphylococcus aureus, Bacillus subtilis, Candida albicans, C. parapsilosis, C. krusei, and C. tropicalis was demonstrated.

Species distribution and susceptibility profile of Candida species in a Brazilian public tertiary hospital

Background. Species identification and antifungal susceptibility tests were carried out on 212 Candida isolates obtained from bloodstream infections, urinary tract infections and dialysis-associated peritonitis, from cases attended at a Brazilian public tertiary hospital from January 1998 to January 2005. Findings. Candida albicans represented 33% of the isolates, Candida parapsilosis 31.1%, Candida tropicalis 17.9%,Candida glabrata 11.8%, and others species 6.2%. In blood culture, C. parapsilosis was the most frequently encountered species (48%). The resistance levels to the antifungal azoles were relatively low for the several species, except for C. tropicalis and C. glabrata. Amphotericin B resistance was observed in 1 isolate of C. parapsilosis. Conclusions. The species distribution and antifungal susceptibility herein observed presented several epidemiological features common to other tertiary hospitals in Latin American countries. It also exhibited some peculiarity, such as a very high frequency of C. parapsilosis both in bloodstream infections and dialysis-associated peritonitis. C. albicans also occurred in an important number of case infections, in all evaluated clinical sources. C. glabrata presented a high proportion of resistant isolates. The data emphasize the necessity to carry out the correct species identification accompanied by the susceptibility tests in all tertiary hospitals. © 2010 Bagagli et al; licensee BioMed Central Ltd.

Avaliação microbiológica da superfície dos implantes ortopédicos metálicos usados para fixar fraturas ósseas

In the majority of cases of bone fracture requiring surgery, orthopedic implants (screw-plate and screw) are used for osteosynthesis and the infections associated with such implants are due to the growth of microorganisms in biofilms. The objective of this study was to identify microorganisms recovered from osteosynthesis implants used to fix bone fractures, to assess the viability of the cells and the ability of staphylococci to adhere to a substrate and to determine their sensitivity/resistance to antimicrobials. After surgical removal, the metal parts of austenitic stainless steel (ASTM F138/F139 or ISO NBR 5832-1/9) were transported to the Laboratory of Clinical Microbiology, washed in buffer and subjected to ultrasonic bath at 40±2 kHz for 5 minutes. The sonicated fluid was used to seed solid culture media and cell viability was assessed under the microscope by with the aid of a fluorescent marker. The production of extracellular polysaccharide by Staphylococcus spp. was investigated by means of adhesion to a polystyrene plate. The profile of susceptibility to antimicrobials was determined by the disk diffusion assay. The most frequently isolated bacteria included coagulase-negative Staphylococcus resistant to erythromycin, clindamycin and oxacillin. Less frequent were Pseudomonas aeruginosa resistant to trimethoprim/sulfamethoxazole and ampicillin, Acinetobacter baumannii resistant to ceftazidime, Enterobacter cloacae resistant to cephalothin, cefoxitin, cefazolin, levofloxacin and ciprofloxacin, Bacillus spp. and Candida tropicalis. The observation of slides by fluorescence microscope showed clusters of living cells embedded in a transparent matrix. The test for adherence of coagulase-negative Staphylococcus to a polystyrene plate showed that these microorganisms produce extracellular polysaccharide. In conclusion, the metal parts were colonized by bacteria related to orthopedic implant infection, which were resistant to multiple antibiotics.

Candida species: Current epidemiology, pathogenicity, biofilm formation, natural antifungal products and new therapeutic options

The incidence of fungal infections has increased significantly, so contributing to morbidity and mortality. This is caused by an increase in antimicrobial resistance and the restricted number of antifungal drugs, which retain many side effects. Candida species are major human fungal pathogens that cause both mucosal and deep tissue infections. Recent evidence suggests that the majority of infections produced by this pathogen are associated with biofilm growth. Biofilms are biological communities with a high degree of organization, in which micro-organisms form structured, coordinated and functional communities. These biological communities are embedded in a self-created extracellular matrix. Biofilm production is also associated with a high level of antimicrobial resistance of the associated organisms. The ability of Candida species to form drugresistant biofilms is an important factor in their contribution to human disease. The study of plants as an alternative to other forms of drug discovery has attracted great attention because, according to the World Health Organization, these would be the best sources for obtaining a wide variety of drugs and could benefit a large population. Furthermore, silver nanoparticles, antibodies and photodynamic inactivation have also been used with good results. This article presents a brief review of the literature regarding the epidemiology of Candida species, as well as their pathogenicity and ability to form biofilms, the antifungal activity of natural products and other therapeutic options. © 2013 SGM.

Identification of Candida species in the clinical laboratory: A review of conventional, commercial, and molecular techniques

In healthy individuals, Candida species are considered commensal yeasts of the oral cavity. However, these microorganisms can also act as opportunist pathogens, particularly the so-called non-albicans Candida species that are increasingly recognized as important agents of human infection. Several surveys have documented increased rates of C. glabrata, C. tropicalis, C. guilliermondii, C. dubliniensis, C. parapsilosis, and C. krusei in local and systemic fungal infections. Some of these species are resistant to antifungal agents. Consequently, rapid and correct identification of species can play an important role in the management of candidiasis. Conventional methods for identification of Candida species are based on morphological and physiological attributes. However, accurate identification of all isolates from clinical samples is often complex and time-consuming. Hence, several manual and automated rapid commercial systems for identifying these organisms have been developed, some of which may have significant sensitivity issues. To overcome these limitations, newer molecular typing techniques have been developed that allow accurate and rapid identification of Candida species. This study reviewed the current state of identification methods for yeasts, particularly Candida species. © 2013 John Wiley & Sons A/S.

Opportunistic microorganisms in individuals with lesions of denture stomatitis

The aim of this study was to isolate, quantify, identify, and compare opportunistic microorganisms (Candida and Staphylococcus genera and Enterobacteriaceae/Pseudomonadaceae families) from prosthesis-fitting surfaces, the hard palate, and mouth rinses of individuals wearing removable maxillary prosthesis with (50) and without (50) lesions of denture stomatitis (DS). The strains were collected and identified using phenotypic, biochemical and molecular tests. The counts of microorganisms were significantly higher in the group of individuals with DS (P < 0.05). C. albicans was the most frequently isolated yeast species in both groups, following by C. tropicalis and C. glabrata. Six isolates were identified as C. dubliniensis. S. aureus and S. epidermidis were the most frequent Staphylococcus species in both groups. Klebsiella pneumoniae was the predominant species in both groups. The association between Candida spp. and bacteria isolated in this study with DS suggests that these microorganisms may play important roles in the establishment and persistence of this disease. © 2013 Elsevier Inc.

Hydroxyurea therapy in sickle cell anemia patients aids to maintain oral fungal colonization balance

Background: The aim of this study was to evaluate the frequency of Candida species and presence of lesions in the oral cavity of patients with sickle cell anemia (SS). Methods: The study included 30 patients diagnosed with sickle cell anemia and taking hydroxyurea for at least 90 days (SS/HU+); and 39 patients with sickle cell anemia and without hydroxyurea therapy (SS/HU-). Two control groups were constituted by healthy individuals matched to the test groups in age, gender, and oral conditions (C/HU+ for SS/HU+ and C/HU- for SS/HU-). Oral clinical examination and anamnesis were performed. Yeasts were collected by oral rinses and identified by API system. Antifungal susceptibility evaluation was performed according to the CLSI methodology. Data obtained for microorganisms counts were compared by Student's t test (SS/HU+ vs. C/HU+ and SS/HU- vs. C/HU-) using MINITAB for Windows 1.4. Significance level was set at 5%. Results: No oral candidosis lesions were detected. Significant differences in yeasts counts were observed between SS/HU- group and the respective control, but there were no differences between SS/HU+ and C/HU+. Candida albicans was the most prevalent species in all groups. Candida famata was observed both in SS and control groups. Candida dubliniensis, Candida glabrata, Candida krusei, Candida tropicalis, Candida pelliculosa, and Candida parapsilosis were observed only in SS groups. Most strains were susceptible to all antifungal agents. Conclusion: Hydroxyurea therapy seems to decrease candidal counts and resistance rate in sickle cell anemia patients. However, further studies should be conducted in the future to confirm this finding. Hydroxyurea therapy in sickle cell anemia patients maintains fungal species balance in oral cavity. © 2013 John Wiley & Sons A/S.

Cytotoxicity of Brazilian plant extracts against oral microorganisms of interest to dentistry

Background: With the emergence of strains resistant to conventional antibiotics, it is important to carry studies using alternative methods to control these microorganisms causing important infections, such as the use of products of plant origin that has demonstrated effective antimicrobial activity besides biocompatibility. Therefore, this study aimed to evaluate the antimicrobial activity of plant extracts of Equisetum arvense L., Glycyrrhiza glabra L., Punica granatum L. and Stryphnodendron barbatimam Mart. against Staphylococcus aureus, Staphylococcus epidermidis, Streptococcus mutans, Candida albicans, Candida tropicalis, and Candida glabrata, and to analyze the cytotoxicity of these extracts in cultured murine macrophages (RAW 264.7).Methods: Antimicrobial activity of plant extracts was evaluated by microdilution method based on Clinical and Laboratory Standards Institute (CLSI), M7-A6 and M27-A2 standards. The cytotoxicity of concentrations that eliminated the microorganisms was evaluated by MTT colorimetric method and by quantification of proinflammatory cytokines (IL-1β and TNF-α) using ELISA.Results: In determining the minimum microbicidal concentration, E. arvense L., P. granatum L., and S. barbatimam Mart. extracts at a concentration of 50 mg/mL and G. glabra L. extract at a concentration of 100 mg/mL, were effective against all microorganisms tested. Regarding cell viability, values were 48% for E. arvense L., 76% for P. granatum L, 86% for S. barbatimam Mart. and 79% for G. glabra L. at the same concentrations. About cytokine production after stimulation with the most effective concentrations of the extracts, there was a significant increase of IL-1β in macrophage cultures treated with S. barbatimam Mart. (3.98 pg/mL) and P. granatum L. (7.72 pg/mL) compared to control (2.20 pg/mL) and a significant decrease of TNF-α was observed in cultures treated with G. glabra L. (4.92 pg/mL), S. barbatimam Mart. (0.85 pg/mL), E. arvense L. (0.83 pg/mL), and P. granatum L. (0.00 pg/mL) when compared to control (41.96 pg/mL).Conclusions: All plant extracts were effective against the microorganisms tested. The G. glabra L. extract exhibited least cytotoxicity and the E. arvense L. extract was the most cytotoxic. © 2013 de Oliveira et al.; licensee BioMed Central Ltd.

Susceptibility profile of a Brazilian yeast stock collection of Candida species isolated from subjects with Candida-associated denture stomatitis with or without diabetes

Objective: This study investigated the susceptibility of 198 clinical isolates of Candida species against caspofungin, amphotericin B, itraconazole, and fluconazole. Study Design: Suspensions of the microorganisms were spread on Roswell Park Memorial Institute (RPMI) agar plates. Etest strips were placed on the plates, and the minimal inhibitory concentration (MIC) was read after incubation (48 h at 37°C). Data were analyzed by a factorial analysis of variance and a 2 × 2 post hoc test (α = .05). Results: C glabrata showed the highest MIC values (P < .001) against caspofungin, itraconazole, and fluconazole. For amphotericin B, the MIC values of C tropicalis and C glabrata (P = .0521) were higher than those of C albicans (P < .001). Itraconazole was the least effective antifungal; 93.3% of the C glabrata isolates, 3.3% of the C albicans, and 1.3% of the C tropicalis were resistant. All microorganisms were susceptible to caspofungin and amphotericin B. Conclusions: Caspofungin and amphotericin B should be recommended as an effective alternative for the management of oral Candida infections when treatment with topical or other systemic drugs has definitely failed. © 2013 Elsevier Inc. All rights reserved.

Photodynamic inactivation of clinical isolates of Candida using Photodithazine

This study evaluated the photodynamic inactivation (PDI) mediated by Photodithazine® (PDZ) against 15 clinical isolates of Candida albicans, Candida glabrata and Candida tropicalis. Each isolate, in planktonic and biofilm form, was exposed to PDI by assessing a range of PDZ concentrations and light emitting diode fluences. Cell survival of the planktonic suspensions was determined by colony forming units (CFU ml-1). The antifungal effects of PDI against biofilms were evaluated by CFU ml-1 and metabolic assay. Data were analyzed by non-parametric tests (α = 0.05). Regardless of the species, PDI promoted a significant viability reduction of planktonic yeasts. The highest reduction in cell viability of the biofilms was equivalent to 0.9 log10 (CFU ml-1) for C. albicans, while 1.4 and 1.5 log10 reductions were obtained for C. tropicalis and C. glabrata, respectively. PDI reduced the metabolic activity of biofilms by 62.1, 76.0, and 76.9% for C. albicans, C. tropicalis, and C. glabrata, respectively. PDZ-mediated PDI promoted significant reduction in the viability of Candida isolates. © 2013 Taylor & Francis.

Evaluation of Syngonanthus nitens (Bong.) Ruhl. extract as antifungal and in treatment of vulvovaginal candidiasis

The purpose of this study was to evaluate the in vitro anticandidal activity of a methanolic extract of Syngonanthus nitens scapes against different Candida species and clinical isolates from patients with vulvovaginal candidiasis (VVC), and its effect in vivo in the treatment of vaginal infection. Chemical characterization of the extract was performed by HPLC-UV analyses and showed the presence of flavones derivatives. The extract was effective against several Candida strains from our collection and species recovered from VVC patients, and was able to inhibit the yeast-hyphal transition. No cytotoxic activity against human female reproductive tract epithelial cells and no hemolytic activity against human red blood cells were observed. In the in vivo model of VVC, we evaluated the efficacy of the intravaginal treatment with a cream containing the extract at doses of 0.5, 1.0 and 2.0%. The treatment eradicated the vaginal fungal burden in infected rats after 8 days of treatment. S. nitens extract could be considered as an effective and non-toxic natural antifungal agent in the treatment of vulvovaginal candidiasis. © 2013 ISHAM.

Pesquisa dos inteins VMA e THRRS em leveduras do gênero Candida isoladas de hemocultura

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Atividade antimicrobiana de Mentha piperita L. sobre leveduras do gênero Candida

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)