Atividade biológica de Cryptococcus neoformans e Cryptococcus gattii provenientes de isolados clínicos e ambientais

Introduction: Cryptococcus neoformans and Cryptococcus gattii are encapsulated basidiomycetous yeasts with worldwide distribution. They cause cryptococcosis with features of systemic infection, affecting the central nervous system, lungs and skin in humans and animals. These fungi present numerous virulence factors that allow them to invade the host and multiply, among which extracellular enzyme capacity and microbial adaptation to different temperatures are worth mentioning. Objective: To evaluate the production of protease and investigate possible differences in thermotolerance and urease activity in clinical and environmental yeast isolates. Material and methods: Culture methods and Pz analysis were applied to assess urease and protease, whereas the optical density method was used to analyze biological activity in thermotolerance. Results: There was no significant results as to microbial growth at the tested temperatures (25º, 37º and 42ºC). It was observed that clinical specimens grew better than environmental ones at elevated temperatures. As to C. neoformans, the moderate production of urease enzyme prevailed in both clinical and environmental isolates within 24h or 48h. Moreover, there was significant production on the seventh day of reading. The best reading time for viewing protease production in both isolates and species was the seventh day: 96% clinical samples and 94% environmental isolates. Conclusion: Further studies are required in order to investigate the virulence factors of C. neoformans and C. gattii cerebrospinal isolates from patients with meningoencephalitis and environmental samples from Sergipe. Furthermore, a higher technical accuracy and statistical precision are indispensable

Mycological study for a management plan of a neotropical show cave (Brazil)

Caves are stable environments with characteristics favoring the development of microorganisms. The allocthonous input of organic matter and microbes into the warm Neotropical caves may favor the development of filamentous fungi, including pathogenic species. Histoplasma capsulatum is a pathogenic species commonly found in caves and associated with bat and bird guano. Many Brazilian caves have been historically visited due to scenic and religious tourism. The objective of this study was to perform a microbiology study for a management plan of a show cave in Brazil, focusing on the presence and distribution of pathogenic and opportunistic fungi in the cave. Statistics analysis was used to verify the influence of touristic activity on air-borne fungi spore load. Fungi were isolated from air and guano in Lapa Nova Cave. Samples were obtained through serial dilution, direct and settle plate techniques. For H. capsulatum, samples were incubated in specific media and conditions. Air-borne fungal spore load was compared prior and during visitation and statistically analyzed. A total of 2,575 isolates from the genera Aspergillus, Calcarisporium, Chaetomium, Cladosporium, Curvularia, Emericella, Eurotium, Fusarium, Geotrichum, Gliocladium, Mucor, Purpureocillium, Paecilomyces, Penicillium, Rhizopus and Trichoderma were identified. Histoplasma capsulatum was not isolated from the cave. Eleven opportunistic species were identified. Significant (p\u3c0.05) variations on fungal richness in the air occurred due to cave visitation. Areas of potential microbiologic risks were indicated and management actions suggested. The results suggest a diverse community inhabiting the cave. Possible opportunistic species should be monitored in show caves and microbiota should always be included in the elaboration of cave management plans. This is the first detailed microbiologic study for a management plan of a show cave in the country. It provides relevant information for future management plans

Occurrence of Aspergillus niger strains on a polychrome cotton painting and their elimination by anoxic treatment

This study aimed to isolate and identify the population of filamentous fungi colonizing a cotton painting, whose conservation status was compromised and showed signs of biodeterioration due to dirt accumulation and microbial metabolism. In addition, microbiological techniques such as cultivation-dependent approach and molecular biology were used to identify microbial populations and to eliminate their metabolic action. For this, the nondestructive anoxic atmosphere technique was used, in which the microbial metabolism was affected by the absence of oxygen. Prior to exposure to an anoxic atmosphere, only one fungal species, Aspergillus niger, was identified at 12 points sampled in the obverse and reverse of the artwork; no fungal species persisted as a result of anoxic treatment. These results showed that exposure to anoxic conditions was effective for the total elimination of isolated fungal strains as well as their spores. In conclusion, this study proved the unprecedented effectiveness of a nondestructive technique for artwork on textile colonized by black fungi species. Thus, this interdisciplinary work involving conservation, microbiology, and chemistry presents a tool to eliminate microorganisms, while maintaining the integrity of artwork and safety of the restorer, that can be applied prior to artwork restoration.The accepted manuscript in pdf format is listed with the files at the bottom of this page. The presentation of the authors' names and (or) special characters in the title of the manuscript may differ slightly between what is listed on this page and what is listed in the pdf file of the accepted manuscript; that in the pdf file of the accepted manuscript is what was submitted by the author

SYNTHESIS AND ANTIFUNGAL ACTIVITY OF PALMITIC ACID-BASED NEOGLYCOLIPIDS RELATED TO PAPULACANDIN D

A series of six new palmitic acid-based neoglycolipids related to Papulacandin D were synthesized in five steps, resulting in good yields, and they were evaluated against Candida spp. All twelve synthetic intermediates were also evaluated. The synthesis involved the initial glycosylation of two phenols (4-hydroxy-3-methoxybenzaldehyde and 3-hydroxybenzaldehyde) via their reaction with peracetylated glucosyl bromide. This was followed by deacetylation with potassium methoxide/metanol solution and the protection of two hydroxyls (C4 and C6 positions) of the saccharide unit as benzilidene acetals (10-11). The next step involved the acylation of the acetal derivatives with palmitic acid, thereby affording a mixture of two isomers mono-acylated at the C2 and C3 positions and a di-acylated product (12-17). After being isolated, each compound was subjected to the removal of the acetal protecting group to yield the papulacandin D analogues 18-23. Three compounds showed low antifungal activity against two species: C. albicans (compounds 7 and 23) and C. tropicalis (compound 17) at 200 µg mL−1

Isolation and identification of Candida species in patients with orogastric cancer: susceptibility to antifungal drugs, attributes of virulence in vitro and immune response phenotype

Submitted by Nuzia Santos ([email protected]) on 2016-08-18T15:58:53Z No. of bitstreams: 1 ve_Sousa_Lourimar_Isolation_CPqRR_2016.pdf: 593875 bytes, checksum: 7f4a0472bd52fe2f7856de91d27401ba (MD5)Approved for entry into archive by Nuzia Santos ([email protected]) on 2016-08-18T16:05:17Z (GMT) No. of bitstreams: 1 ve_Sousa_Lourimar_Isolation_CPqRR_2016.pdf: 593875 bytes, checksum: 7f4a0472bd52fe2f7856de91d27401ba (MD5)Made available in DSpace on 2016-08-18T16:05:17Z (GMT). No. of bitstreams: 1 ve_Sousa_Lourimar_Isolation_CPqRR_2016.pdf: 593875 bytes, checksum: 7f4a0472bd52fe2f7856de91d27401ba (MD5) Previous issue date: 2016Universidade Federal de Minas Gerais. Instituto de Ciencias Biologicas. Departamento de Microbiologia. Laboratorio de Micologia. Belo Horizonte, MG, Brasil/Universidade Vale do Rio Doce. Laboratorio de Microbiologia. Governador Valadares, MG, Brasil.Universidade Federal de Minas Gerais. Instituto de Ciencias Biologicas. Departamento de Microbiologia. Laboratorio de Microbiologia. Belo Horizonte, MG, Brasil.Universidade CEUMA. Laboratorio de Microbiologia. São Luis, MA, Brasil.Universidade Federal de Minas Gerais. Instituto de Ciencias Biologicas. Departamento de Microbiologia. Laboratorio de Microbiologia. Belo Horizonte, MG, Brasil.Universidade Federal do Maranhão. Hospital Universitario. São Luis, MA, Brasil.Fundação Oswaldo Cruz. Centro de Pesquisa Rene Rachou. Laboratorio de Biomarcadores de Diagnostico e Monitoramento. Belo Horizonte, MG, Brasil.Universidade Vale do Rio Doce. Laboratorio de Microbiologia. Governador Valadares, MG, Brasil.Universidade Federal de Minas Gerais. Instituto de Ciencias Biologicas. Laboratorio de Microbiologia Oral e Anaerobica. Belo Horizonte, MG, Brasil.Core Experts Oncologia. Governador Valadares, MG, BrasilLaboratorio Alvarenga. Governador Valadares, MG, BrasilUniversidade Federal de Minas Gerais. Instituto de Ciencias Biologicas. Departamento de Microbiologia. Laboratorio de Micologia. Belo Horizonte, MG, Brasil.Universidade Federal de Minas Gerais. Instituto de Ciencias Biologicas. Departamento de Microbiologia. Laboratorio de Virologia Basica e Aplicada. Belo Horizonte, MG, Brasil.Federal Universidade Federal do Espirito Santo. Departamento de Patologia. Laboratorio de Bacteriologia Geral e Clinica. Centro de Ciencias da Saude. Vitória, ES, Brasil.Universidade Federal de Minas Gerais. Instituto de Ciencias Biologicas. Departamento de Microbiologia. Laboratorio de Micologia. Belo Horizonte, MG, Brasil.Background: Because of the inherent immunosuppression of cancer patients opportunistic infections by Candida spp, occur frequently. This study aimed to identify Candida species in the oral mucosa of 59 patients with orogastric cancer (OGC) and to analyze the immunological phenotype of these patients. Methods: The yeasts were identified by MALDI-TOF mass spectrometry (MS). For all isolates, we performed phospholipases and proteinases assays, in vitro adherence to buccal epithelial cells (BEC), minimum inhibitory concentration of antifungal drugs and determined the cytokine profile by Cytometric Bead Array flow citometry assay. Results: C. albicans was the most prevalent species in OGC patients (51.6 %) and control group (66.7 %). Candida spp. strains isolated from OGC patients exhibited better adherence to BEC (p = 0.05) than did the control group. Phospholipases production by Candida strains from OGC patients was lower (51.6 %) than in the control group (61.9 %). Proteinases were detected in 41.9 % and 4.8 % of the yeasts from OGC patients and control group, respectively. Significant differences were found in the serum of OGC patients compared to the control group for IL-2, IL-10, TNF-α, IFN-γ and IL-17. Conclusions: The results of this work suggest increased virulence of yeasts isolated from OGC patients and, that this may interfere with the immune phenotype

Butenafine and analogues: An expeditious synthesis and cytotoxicity and antifungal activities

The incidence of fungal infections is considered a serious public health problem worldwide. The limited number of antimycotic drugs available to treat human and animal mycosis, the undesirable side effects and toxicities of the currently available drugs, and the emergence of fungal resistance emphasizes the urgent need for more effective antimycotic medicines. In this paper, we describe a rapid, simple, and efficient synthetic route for preparation of the antifungal agent butenafine on a multigram scale. This novel synthetic route also facilitated the preparation of 17 butenafine analogues using Schiff bases as precursors in three steps or less. All the synthesized compounds were evaluated against the yeast, Cryptococcus neoformans/C. gattii species complexes and the filamentous fungi Trichophyton rubrum and Microsporum gypseum. Amine 4bd, a demethylated analogue of butenafine, and its corresponding hydrochloride salt showed low toxicity in vitro and in vivo while maintaining inhibitory activity against filamentous fungi. Keywords: Antifungal activity, Butenafine, Microwave-assisted synthesis, Multigram-scale synthesis, Schiff base, Trichophyton rubru

Fluconazole Alters the Polysaccharide Capsule of Cryptococcus gattii and Leads to Distinct Behaviors in Murine Cryptococcosis

Abstract Cryptococcus gattii is an emergent human pathogen. Fluconazole is commonly used for treatment of cryptococcosis, but the emergence of less susceptible strains to this azole is a global problem and also the data regarding fluconazole-resistant cryptococcosis are scarce. We evaluate the influence of fluconazole on murine cryptococcosis and whether this azole alters the polysaccharide (PS) from cryptococcal cells. L27/01 strain of C. gattii was cultivated in high fluconazole concentrations and developed decreased drug susceptibility. This phenotype was named L27/01 F , that was less virulent than L27/01 in mice. The physical, structural and electrophoretic properties of the PS capsule of L27/01 F were altered by fluconazole. L27/ 01 F presented lower antiphagocytic properties and reduced survival inside macrophages. The L27/01 F did not affect the central nervous system, while the effect in brain caused by L27/01 strain began after only 12 hours. Mice infected with L27/ 01 F presented lower production of the pro-inflammatory cytokines, with increased cellular recruitment in the lungs and severe pulmonary disease. The behavioral alterations were affected by L27/01, but no effects were detected after infection with L27/01 F . Our results suggest that stress to fluconazole alters the capsule of C. gattii and influences the clinical manifestations of cryptococcosis