Environmental isolation, biochemical identification, and antifungal drug susceptibility of Cryptococcus species

Introduction The incidence of opportunistic fungal infections has increased in recent years and is considered an important public health problem. Among systemic and opportunistic mycoses, cryptococcosis is distinguished by its clinical importance due to the increased risk of infection in individuals infected by human immunodeficiency virus. Methods To determine the occurrence of pathogenic Cryptococcus in pigeon excrement in the City of Araraquara, samples were collected from nine environments, including state and municipal schools, abandoned buildings, parks, and a hospital. The isolates were identified using classical tests, and susceptibility testing for the antifungal drugs (fluconazole, itraconazole, voriconazole, and amphotericin B) independently was also performed. After collection, the excrement samples were plated on Niger agar and incubated at room temperature. Results A total of 87 bird dropping samples were collected, and 66.6% were positive for the genus Cryptococcus. The following species were identified: Cryptococcus neoformans (17.2%), Cryptococcus gattii (5.2%), Cryptococcus ater (3.5%), Cryptococcus laurentti (1.7%), and Cryptococcus luteolus (1.7%). A total of 70.7% of the isolates were not identified to the species level and are referred to as Cryptococcus spp. throughout the manuscript. Conclusions Although none of the isolates demonstrated resistance to antifungal drugs, the identification of infested areas, the proper control of birds, and the disinfection of these environments are essential for the epidemiological control of cryptococcosis.

Molecular Identification of Nocardia Isolates from Clinical Samples and an Overview of Human Nocardiosis in Brazil

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Comparação entre método bioquímico e reação em cadeia de polimerase para identificação de Lactobacillus spp., isolados de aves

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

Caracterização fenotípica de leveduras isoladas da mucosa vaginal em mulheres adultas

OBJETIVO: caracterizar fenotipicamente leveduras isoladas do conteúdo vaginal de 223 mulheres adultas, sintomáticas (S) e assintomáticas (A) para vulvovaginite, e determinar os indicadores clínicos que possivelmente levam ao surgimento de sinais e sintomas relacionados ao acometimento da mucosa por essa patologia. MÉTODOS: inicialmente foi aplicado um questionário, com questões abertas e fechadas, sobre dados clínicos epidemiológicos. Logo, ocorreu o diagnóstico micológico com semeadura em meio Chrom Agar Candida, identificação micromorfológica e bioquímica. Métodos específicos para detecção de fatores de virulência, proteinase e fosfolipase foram empregados. A análise estatística das variáveis foi estabelecida utilizando os testes χ2 e χ2 de Pearson. RESULTADOS: Candida albicans foi a espécie mais prevalente (87%, S e 67%, A), seguida de Candida glabrata (4%, S e 17%, A). O número de mulheres que referiram adoção de anticoncepcionais foi mais alto entre as sintomáticas, 77%. Nos dois grupos estudados, em torno de 87% apresentaram ciclos menstruais regulares, 57% das mulheres eram casadas com idade entre 30 a 40 anos. em relação a práticas sexuais, houve para parte das pacientes, concomitância entre os hábitos, anal, oral e vaginal. em relação à fosfolipase, apenas Candida albicans produziu este fator de virulência em 37,5%. A proteinase foi detectada em Candida albicans, Candida glabrata e Candida parapsilosis. Esse último fator de virulência esteve associado, principalmente, a isolados de pacientes sintomáticas. CONCLUSÕES: a colonização e infecção da mucosa vaginal por levedura é real com diversas espécies de Candida presentes. No entanto, Candida albicans se destaca como espécie prevalente em mucosa vaginal de mulheres adultas. Fica evidente a emergência de espécies de Candida não albicans, algumas com resistência intrínseca aos azólicos, tais como Candida glabrata, Candida parapsilosis, Candida tropicalis, e Candida guillermondii, o que pode ser explicado pelo uso inadequado de medicamentos e tratamento empírico.

Identificação de Enterobacter spp. através de técnicas convencionais e do sistema semi-automatizado Microscan (Autoscan-4)

A hundred and ninety-five (195) strains of Enterobacter spp., isolated from diverse clinical specimens - urine, feces, cateter, blood, wound, tracheal aspirate, vaginal fluid - were submitted to the conventional identification by biochemical tests, and were also submitted to the identification by panels NegCombo 20 of the system automated MicroScan - AutoScan- 4 (Dade Behring Inc., West Sacramento, CA, USA). The samples were from patients of the Clinical Laboratory from the School of Pharmacy and Biochemistry of UNOESTE, Presidente Prudente, SP, and from patients hospitalized at the University Hospital Domingos Leonardo Cerávolo, UNOESTE. Of the total of strains tested, 191 (97.9%) presented agreement between the two approaches utilized and 4 strains (2.1%) presented identification disagreement, that is, the genus identified was different in each approach. By this study, the conclusion is that both the approaches utilized for the identification presented advantages and disadvantages related to the cost, facility of execution, quickness, reliability and some other characteristics. Even so, our results showed that conventional methods represent a reliable tool for Enterobacter identification.

Recovery of mutans streptococci on MSB, SB-20 and SB-20M agar media

Objective: The recovery of mutans streptococci in saliva and dental biofilm samples depends, in part, on the culture medium used. In this study, we compared (i) the culture media Sucrose-Bacitracin agar (SB-20), Modified SB-20 (SB-20M) and Mitis Salivarius Bacitracin agar (MSB) in the count of colony forming units (cfu) of mutans streptococci and (ii) in the morphological and biochemical differentiation between Streptococcus mutans and Streptococcus sobrinus. Design: Samples of non-stimulated saliva from 20 children were plated on SB-20, SB-20M and MSB, and incubated in microaerophilia at 37 °C for 72 h. Identification of microorganisms was based on analysis of colony morphology under stereomicroscopy. The biochemical identification of colonies was done by biochemical tests using sugar fermentation, resistance to bacitracin and hydrogen peroxide production. Results: There was no significant difference (p > 0.05) in the number of cfu of mutans streptococci recovered on SB-20 and SB-20M agar. Comparing the media, SB-20 and SB-20M yielded a larger number of mutans streptococci colonies (p < 0.05) and were more effective than MSB in the identification of S. sobrinus (p < 0.05), but not of S. mutans (p > 0.05). Conclusion: There was no significant difference between SB-20 and SB-20M culture media in the count of mutans streptococci, demonstrating that the replacement of sucrose by coarse granular cane sugar did not alter the efficacy of the medium. Compared with MSB, SB-20 and SB-20M allowed counting a larger number of mutans streptococci colonies and a more effective morphological identification of S. sobrinus. © 2012 Elsevier Ltd.

Análise comparativa de métodos de diagnóstico para linfadenite caseosa em ovinos sintomáticos e assintomáticos

Pós-graduação em Ciência Animal - FMVA

Desinfecção de próteses totais por micro-ondas: efeito da frequência de irradiação no tratamento da estomatite protética

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

Validação de técnicas moleculares para o diagnóstico de bactérias em peixes, visando redução de tempo e custo

Pós-graduação em Microbiologia Agropecuária - FCAV

Subclinical diagnosis of Caseous lymphadenitis based on Elisa in Sheep from Brazil

Caseous lymphadenitis (CLA), caused by Corynebacterium pseudotuberculosis, is a chronic contagious disease that affects small ruminants and still remains an important problem for many lamb-producing countries. Animals are considered clinically infected when occurs abscesses in superficial lymph nodes. Visceral or internal form can coexist which no apparent clinical signs of infection are seen. The best procedure to avoid spread of the disease is elimination of infected animals. However, as the chronic and subclinical nature of the infection of CLA alternative methods are required for detection and screening. In this study, we described the performance of indirect Enzyme-Linked Immunosorbent Assay (ELISA) for diagnosis of CLA in asymptomatics sheep. Also, test culture and biochemical identification were achieved to confirm CLA infection. The serological diagnostic was performed in sheep symptomatics (n=50) and asymptomatics (n=374) from nine flocks. Analysis reported high positivity of 71% for ELISA in 85% of asymptomatic animal for CLA with a sensitivity of 88% and specificity of 31%. Results from ELISA test in asymptomatic animals against culture for caseous lymphadenitis were more specific (97%) and permitted to exclude healthy animals without symptoms. This study concluded that C. pseudotuberculosis infection could be widely disseminated in sheep flocks in Northwestern region of the state of São Paulo, Brazil and only one screening test is not enough. The association with indirect ELISA test and culture could better indicate the real problem of CLA in sheep flocks.

Identification of Staphylococcus saprophyticus isolated from patients with urinary tract infection using a simple set of biochemical tests correlating with 16S-23S interspace region molecular weight patterns

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

Identification of Mycobacteria by Thin Layer Chromatographic Analysis of Mycolic Acids and Conventional Biochemical Method: Four Years of Experience

Mycolic acids analysis by thin-layer chromatography (TLC) has been employed by several laboratories worldwide as a method for fast identification of mycobacteria. This method was introduced in Brazil by our laboratory in 1992 as a routine identification technique. Up to the present, 861 strains isolated were identified by mycolic acids TLC and by standard biochemical tests; 61% out of these strains came as clinical samples, 4% isolated from frogs and 35% as environmental samples. Mycobacterium tuberculosis strains identified by classical methods were confirmed by their mycolic acids contents (I, III and IV). The method allowed earlier differentiation of M. avium complex - MAC (mycolic acids I, IV and VI) from M. simiae (acids I, II and IV), both with similar biochemical properties. The method also permitted to distinguish M. fortuitum (acids I and V) from M. chelonae (acids I and II) , and to detect mixed mycobacterial infections cases as M. tuberculosis with MAC and M. fortuitum with MAC. Concluding, four years experience shows that mycolic acids TLC is an easy, reliable, fast and inexpensive method, an important tool to put together conventional mycobacteria identification methods.

The Identification and Biochemical Properties of the Catalytic Specificity of a Serine Peptidase Secreted by Aspergillus fumigatus Fresenius

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

Structural and Biochemical Characterization of Peroxiredoxin Q beta from Xylella fastidiosa CATALYTIC MECHANISM and HIGH REACTIVITY

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

Identification of a novel melittin isoform from Africanized Apis mellifera venom

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