Fungemia in a university hospital: an epidemiological approach

INTRODUCTION: Fungemia corresponds to the isolation of fungi in the bloodstream and occurs mostly in immunosuppressed patients. The early diagnosis and treatment of these infections are relevant given the serious threat to the affected patients and possible spread to other organs, often becoming fatal. The growing number of fungemia associated with poor prognosis resulted in this research aiming to diagnose and assess the epidemiological aspects of hematogenous infections by fungi. METHODS: The study included 58 blood samples collected within a 1-year period, from patients at the Hospital das Clinicas, Federal University of Pernambuco, by venipuncture in vacuum tubes. Blood samples were processed for direct examination and culture and identification, conducted by observing the macroscopic and microscopic characteristics, as well as physiological characteristics when necessary. RESULTS: Eight (13.8%) episodes of fungemia were identified, accounting for the total sample, and these pathogens were Candida, Histoplasma, Trichosporon, Cryptococcus, and a dematiaceous fungus. C. albicans was the prevalent species, accounting for 37.5% of the cases. Most affected patients were adult males. There was no predominance for any activity, and the risk of acquired immunodeficiency syndrome was the underlying pathology most often cited. CONCLUSIONS: The isolation of fungi considered as emergent species, such as C. membranifaciens and dematiaceous species, highlights the importance of epidemiological monitoring of cases of fungemia in immunocompromised patients, as the therapy of choice depends on the knowledge of the aethiological agent.

Clinical and mycological study of scalp white piedra in the State of Paraíba, Brazil

White piedra is a superficial mycoses characterized by nodules on the hair shaft, caused by the basidiomycetous yeasts. In the present study, clinical and mycological findings of scalp white piedra caused by Trichosporon spp. are related. Twenty three cases of scalp white piedra were observed with a high incidence in women (87%) and preschool children from 2 to 6 (74%) years old. These groups presented a relationship of dependence with this infection. Despite the low socio-economic status, poor standards of hygiene, (48% of the patients) as well as the fact that 30.4% of the children shared the same nursery, these factors were not significant for the transmission of the mycosis. These were the first reports of scalp white piedra in João Pessoa city, Paraíba, Brazil.

Antifungal activity of propolis extract against yeasts isolated from onychomycosis lesions

The aim of this study was to determine the in vitro activity of propolis extract against 67 yeasts isolated from onychomycosis in patients attending at the Teaching and Research Laboratory of Clinical Analysis of the State University of Maringá. The method used was an adaptation made from the protocol approved by the National Committee for Clinical Laboratory Standards. The yeasts tested were: Candida parapsilosis 35%, C. tropicalis 23%, C. albicans 13%, and other species 29%. The propolis extract showed excellent performance regarding its antifungal activity: the concentration capable of inhibiting the all of the yeasts was 5 × 10-2 mg/ml of flavonoids and 2 × 10-2 mg/ml of flavonoids stimulated their cellular death. Trichosporon sp. were the most sensitive species, showing MIC50 and MIC90 of 1.25 × 10-2 mg/ml of flavonoids, and C. tropicalis was the most resistant, with CFM50 of 5 × 10-2 mg/ml of flavonoids and MFC90 of 10 × 10-2 mg/ml. In view of the fact that propolis is a natural, low cost, non-toxic product with proven antifungal activity, it should be considered as another option in the onychomycosis treatment.

Morphological and biochemical characterization of the aetiological agents of white piedra

The Trichosporon genus is constituted by many species, of which Trichosporon ovoides and Trichosporon inkin are the causative agents of white piedra. They can cause nodules in genital hair or on the scalp. At present, Brazilian laboratory routines generally do not include the identification of the species of Trichosporon genus, which, although morphologically and physiologically distinct, present many similarities, making the identification difficult. The aim of this study was to identify the aetiological agents at the species level of white piedra from clinical specimens. Therefore, both the macro and micro morphology were studied, and physiological tests were performed. Trichosporon spp. was isolated from 10 clinical samples; T. ovoides was predominant, as it was found in seven samples, while T. inkin was identified just in two samples. One isolate could not be identified at the species level. T. inkin was identified for the first time as a white piedra agent in the hair shaft on child under the age of 10.

Identification and antifungal susceptibility of a large collection of yeast strains isolated in Tunisian hospitals.

Abstract In this study, matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) was used as a rapid method to identify yeasts isolated from patients in Tunisian hospitals. When identification could not be exstablished with this procedure, sequencing of the internal transcribed spacer with 5.8S ribosomal DNA (rDNA) (ITS1-5.8S-ITS2) and D1/D2 domain of large-subunit (LSU rDNA) were employed as a molecular approach for species differentiation. Candida albicans was the dominant species (43.37% of all cases), followed by C. glabrata (16.55%), C. parapsilosis (13.23%), C. tropicalis (11.34%), C. dubliniensis (4.96%), and other species more rarely encountered in human diseases such as C. krusei, C. metapsilosis, C. lusitaniae, C. kefyr, C. palmioleophila, C. guilliermondii, C. intermedia, C. orthopsilosis, and C. utilis. In addition, other yeast species were obtained including Saccharomyces cerevisiae, Debaryomyces hansenii (anamorph known as C. famata), Hanseniaspora opuntiae, Kodamaea ohmeri, Pichia caribbica (anamorph known as C. fermentati), Trichosporon spp. and finally a novel yeast species, C. tunisiensis. The in vitro antifungal activities of fluconazole and voriconazole were determined by the agar disk diffusion test and Etest, while the susceptibility to additional antifungal agents was determined with the Sensititre YeastOne system. Our results showed low incidence of azole resistance in C. albicans (0.54%), C. tropicalis (2.08%) and C. glabrata (4.28%). In addition, caspofungin was active against most isolates of the collection with the exception of two K. ohmeri isolates. This is the first report to describe caspofungin resistant isolates of this yeast.

Mycoflora and aflatoxigenic species in derivatives of milled rice

Thirty samples of rough rice stored for 6, 12 and 24 months in government authorized warehouses of the state of Rio Grande do Sul, Brazil, were simultaneously collected. After milling of the product, 90 samples (30 of polished rice, 30 of rice bran and 30 of rice hull) were evaluated for their mycoflora, aflatoxigenic species and aflatoxin contamination. The following fungi, listed in decreasing order of frequency, were isolated on Potato-Dextrose Agar: Aspergillus spp., Nigrospora spp., Penicillium spp.; Fusarium spp.; Mucor spp.; Cladosporium spp.; Trichosporon spp. and non-sporulated fungi. The degree of fungal contamination (colony forming units per gram of product) was lowest in polished rice, increasing progressively in samples of rice bran and rice hull. Among the Aspergillus species, A. flavus and A. candidus were isolated most frequently. Of the A. flavus isolates, 52.6% strains were found to be toxigenic and produced only Group B aflatoxins. Analysis of the 90 samples did not reveal the presence of aflatoxins in the rice derivatives.

Characterization of the Phenol Monooxygenase Gene from Chromobacterium violaceum: Potential Use for Phenol Biodegradation

In this work, the biodegradation mechanism of phenol and sub products (such as catechol and hydroquinone) in Chromobacterium violaceum was investigated by cloning and molecular characterization of a phenol monooxygenase gene in Escherichia coli. This gene (Cvmp) is very similar (74 and 59% of similarity and identity, respectively) to the ortholog from Ralstonia eutropha, bacteria capable of utilizing phenol as the sole carbon source. The phenol biodegradation ability of E. coli recombinant strains was tested by cell-growth in a minimal medium containing phenol as the sole source of carbon and release of intermediary metabolites (catechol and hydroquinone). Interestingly, during the growth of these strains on phenol, catechol, and hydroquinone accumulated transiently in the medium. These metabolites were further analyzed by HPLC. These results indicated that phenol can be initially orto or para hydroxylated to produce cathecol or hydroquinone, respectively, followed by meta-cleavage of aromatic rings. To verify this information, the metabolites obtained from HPLC were submitted to LC/MS to confirm their chemical structure, thereby indicating that the recombinant strains utilize two different routes simultaneously, leading to different ring-fission substrates for the metabolism of phenol. (C) KSBB

Cryptococcus haglerorum, sp nov., an anamorphic basidiomycetous yeast isolated from nests of the leaf-cutting ant Atta sexdens

A yeast strain (CBS 8902) was isolated from the nest of a leaf-cutting ant and was shown to be related to Cryptococcus humicola. Sequencing of the D1/D2 region of the 26S ribosomal DNA and physiological characterization revealed a separate taxonomic position. A novel species named Cryptococcus haglerorum is proposed to accommodate strain CBS 8902 that assimilates n-hexadecane and several benzene compounds. Physiological characteristics distinguishing the novel species from some other members of the C. humicola complex are presented. The phylogenetic relationship of these strains to species of the genus Trichosporon Behrend is discussed.

Monitoramento de fungos anemófilos e de leveduras em unidade hospitalar

OBJETIVO: Monitorar e caracterizar fungos anemófilos e leveduras de fontes bióticas e abióticas de uma unidade hospitalar. MÉTODOS: As coletas foram realizadas mensalmente e em dois períodos, do centro cirúrgico e unidades de terapia intensiva adulto e neonatal em hospital de Araraquara, Estado de São Paulo. Para coleta de fungos anemófilos foi utilizado amostrador tipo Andersen de simples estágio. A pesquisa de leveduras foi feita das mãos e de orofaringe de profissionais de saúde, bem como de superfícies de leitos e de maçanetas das áreas críticas. RESULTADOS: Foram recuperados do centro cirúrgico 32 gêneros de fungos anemófilos e 31 das unidades de terapia intensiva. Os gêneros mais freqüentemente isolados foram Cladophialophora spp., Fusarium spp., Penicillium spp., Chrysosporium spp. e Aspergillus spp. Durante o período de estudo, houve reforma e implantação de uma unidade dentro do hospital, que coincidiu com o aumento na contagem de colônias de Cladophialophora spp., Aspergillus spp. e Fusarium spp. Leveduras foram encontradas em 39,4% dos profissionais de saúde (16,7% das amostras dos espaços interdigitais, 12,1% do leito subungueal e 10,6% da orofaringe) e, em 44% das amostras do mobiliário, com predomínio do gênero Candida (C. albicans, C. guilliermondii, C. parapsilosis e C. lusitaniae) seguido por Trichosporon spp. CONCLUSÕES: Observou-se número relativamente elevado de fungos anemófilos (potencialmente patogênicos) em áreas especiais e níveis expressivos de leveduras em fontes bióticas e abióticas. O monitoramento microbiológico ambiental deve ser realizado, principalmente em salas especiais com pacientes imunocomprometidos, sujeitos à exposição de patógenos do meio ambiente, assim como, advindos de profissionais de saúde.

Species diversity of yeast in oral colonization of insulin-treated diabetes mellitus patients

The aim of this study was to investigate oral yeast colonization, antifungal susceptibility and strain diversity in insulin-dependent diabetes mellitus patients (175), as well as to evaluate the influence of dental prostheses. Oral rinse samples were cultured on selective media, in order to isolate, count and identify the yeasts recovered. More than half of the diabetic subjects (53%) carried significant amounts of Candida cells in the buccal cavity and these organisms were recovered at higher densities in diabetics wearing dentures. A total of 93 yeast strains were isolated from these patients, including: Candida spp. (n = 89); Pichia (n = 02); Trichosporon (n = 1), and Geotrichum (n = 1). C. albicans represented 56% of these strains, non-albicans Candida 39.8%, and other genera of yeast 4.3%. C. albicans was prevalent, followed by C. parapsilosis, C. tropicalis, C. glabrata, C. krusei, C. rugosa and C. guilliermondii. Agar disk-diffusion tests of the susceptibility of non-albicans Candida and other genera of yeast to fluconazole showed resistance in 21.9%, mainly in C. rugosa (100%), C. glabrata (57%) and C. krusei (50%). Local oral factors, such as the presence of dentures, in association with diabetes, seemed to have the effect of increasing the amount and variety of Candida species in the oral cavities, mainly those with lower drug susceptibilities.

Oral colonization by yeasts in hiv-positive patients in Brazil

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

White piedra and pediculosis capitis in the same patient

Piedra branca é micose superficial causada pelo gênero Trichosporon e caracterizada por nódulos aderidos à haste do pelo. Pediculose capitis é causada pelo Pediculus humanus var. capitis pertencente à subordem Anoplura. Enquanto que a pediculose é enfermidade comum, relatos clínicos de piedra branca são raros. Técnicas de biologia molecular identificaram o agente de piedra branca do presente relato como T.inkin. Os autores apresentam associação de ambas as infestações no mesmo paciente para salientar seus aspectos clínicos distintos.

Yeasts associated with nests of the leaf-cutting ant Atta sexdens rubropilosa Forel, 1908

A total of 137 yeasts associated with the leaf-cutting ant Atta sexdens rubropilosa Forel, 1908 were characterized, being selected 93 for analysis. Twenty four species belonging to seven genera(Candida, Cryptococcus, Rhodotorula, Sporobolomyces, Tremella, Trichosporon, Pichia) were isolated from the different analysed material. The genus Candida was widely distributed, with C. homilentoma, C. colliculosa-like, C. famata and C. colliculosa being the most prevalent. A few isolates did not fit the standard descriptions and probably some of them could be new biotypes or even new species. Three strains of black yeasts were also isolated, and four others were identified as being Candida spp. The effective number of yeast species was higher in newer sponge. The origin, distribution and relative importance of these microorganisms for the ants are discussed.

Occurrence of killer yeasts in leaf-cutting ant nests

Killer activity was screened in 99 yeast strains isolated from the nests of the leaf-cutting ant Atta sexdens against 6 standard sensitive strains, as well as against each other. Among this yeast community killer activity was widespread since 77 strains (78 %) were able to kill or inhibit the growth of at least one standard strain or nest strain. Toxin production was observed in representatives of all the studied genera including Aureobasidium, Rhodotorula, Tremella and Trichosporon, whose killer activity has not yet been described.

Descrição da diversidade de fungos associados a formigas Attini por método independente de cultivo

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