Different scenarios for Candida parapsilosis fungaemia reveal high numbers of mixed C-parapsilosis and Candida orthopsilosis infections

Nosocomial fungal bloodstream infections (BSI) are increasing significantly in hospitalized patients and Candida parapsilosis has emerged as an important pathogen responsible for numerous outbreaks. The objective of this study was to evaluate C. parapsilosis sensu lato infection scenarios, regarding species distribution and strain relatedness. One hundred isolates of C. parapsilosis sensu lato derived from blood cultures and catheter tips were analysed by multiplex microsatellite typing and by sequencing D1/D2 regions of the ribosomal DNA. Our results indicate that 9.5 % of patients presented infections due to C. parapsilosis and Candida orthopsilosis, 57.1 % due to C. parapsilosis, 28.3 % due to C. orthopsilosis and 4.8% due to Candida metapsilosis. Eighty per cent of the C. parapsilosis BSIs were due to a single strain that was also identified in the catheter, but in 10% of the cases C. parasilosis was identified in the catheter but the BSI was due to C. orthopsilosis. There is a significant probability that C. parapsilosis isolates collected from the same patient at more than 3 months interval are of different strains (P=0.0179). Moreover, several isolates were identified persistently in the same hospital, infecting six different patients. The incidence of polyfungal BSI infections with C. parapsilosis and C. orthopsilosis is reported herein for the first time, emphasizing the fact that the species identified in the catheter is not always responsible for the BSI, thus impacting the treatment strategy. The observation that strains can remain in the hospital environment for years highlights the possible existence of reservoirs and reinforces the need for accurate genotyping tools, such as the markers used for elucidating epidemiological associations and detecting outbreaks.Financial support was provided by CAPES Foundation (BEX 19194/12-9), Ministry of Education of Brazil, Brasilia (DF 70.040-020), by FEDER through POFC-COMPETE and by Portuguese funds from FCT (PEst-OE/BIA/UI4050/2014). R.M.Z.-O. is supported in part by CNPq (350338/2000-0) and FAPERJ E (26/103.157/2011). We are grateful to Ronaldo Rozembaun from HSE and SAM and Andrea Pussenti Derossi from HUPE for providing the Candida isolates and technical assistance in sampling. Automated sequencing was done using the genomic platform/DNA sequencing platform at Fundacao Oswaldo Cruz, PDTIS/FIOCRUZ (RPT01A), Brazil. The authors declare that they have no conflict of interest.info:eu-repo/semantics/publishedVersio

HISTOPLASMIN SURVEY IN HIV-POSITIVE PATIENTS: RESULTS FROM AN ENDEMIC AREA IN NORTHEASTERN BRAZIL

SUMMARY Background. Disseminated histoplasmosis is common in AIDS patients with advanced immunosuppression in Ceará, Northeastern Brazil. The goal of this study was to determine the prevalence of Histoplasma infection in patients with HIV/AIDS living in Fortaleza, the capital of Ceará. Methods. Intradermal tests with histoplasmin (mycelial phase) were performed in 161 HIV patients with CD4 ≥ 350 cells/mm 3 . Evidence of recent illness was evaluated with immunodiffusion (ID) tests in 76 of these individuals. Results. A total of 11.8% of patients reacted to histoplasmin and 2.63% had ID test positive to Histoplasma. The presence of mango trees (Mangifera indica) in the patient neighborhood (OR = 2.870; 95% CI = 1.081-7.617; p = 0.040) and past activity involving soil (OR = 2.834; 95% CI = 1.045-7.687; p = 0.045) or visits to a farm (OR = 3.869; 95% CI = 1.189-12.591; p = 0.033) were significantly associated with Histoplasma infection. Conclusions. Patients with HIV living in Fortaleza have an expressive prevalence of infection with Histoplasma.RESUMO Introdução: Histoplasmose disseminada ocorre com grande frequência em pacientes com aids e imunossupressão avançada no Ceará, Brasil. O objetivo deste artigo é determinar a prevalência da infecção por Histoplasma em pacientes com HIV/aids residentes em Fortaleza capital. Métodos. Testes intradérmicos com histoplasmina (fase micelial), foram realizados em 161 pacientes com CD4 ≥ 350 células/mm 3 . Doença recente foi estudada por imunodifusão em 76 desses indivíduos. Resultados. Reagiram à histoplasmina, 11,8% dos pacientes e à imunodifusão para Histoplasma: 2,63%. A presença da árvore mangueira (Mangifera indica) na vizinhança (OR = 2,870; IC 95% = 1,081-7,617; p = 0,040), atividade com o solo no passado (OR = 2,834; IC 95% = 1,045-7,687, p = 0,045) e visitar sítio no passado (OR = 3,869; IC 95% = 1,189-12,591; p = 0,033); foram significativamente associados com positividade para o teste. Conclusões. Pacientes com HIV que vivem em Fortaleza apresentam uma prevalência expressiva de infecção por Histoplasma

Evaluation of T3B fingerprinting for identification of clinical and environmental Sporothrix species

In this study, PCR fingerprinting using the universal primer T3B was applied to distinguish among clinical and environmental species of the Sporothrix complex, Sporothrix brasiliensis, S. globosa, S. mexicana, S. pallida, S. luriei and S. schenckii sensu stricto. The T3B fingerprinting generated clearly distinct banding patterns, allowing the correct identification of all 43 clinical and environmental isolates at the species level, what was confirmed by partial calmodulin gene sequence analyses. This technique is reproducible and provides the identification of all species of the Sporothrix complex with sufficient accuracy to be applied in clinical mycology laboratories as well as in epidemiological studies in order to obtain a better understanding of the epidemiology of sporotrichosis.This study was approved by the Research Ethics Committee of IPEC/Fiocruz. Financial support for this work was provided by FAPERJ (Grant Proc. E-26/111.619/2008). R.M.Z.O. is in part supported by CNPq 350338/2000-0. M.M.E.O. was supported in part by a grant from CAPES 2445/11-5 and CAPES-PNPD for his work at CBMA, Universidade do Minho, Braga, PT.info:eu-repo/semantics/publishedVersio

Molecular identification of Sporothrix species involved in the first familial outbreak of sporotrichosis in the state of Espírito Santo, southeastern Brazil

Sporotrichosis is a widespread subcutaneous mycosis caused by the dimorphic fungi now known as the Sporothrix schenckii complex. This complex is comprised of at least six species, including Sporothrix albicans, Sporothrix brasiliensis, Sporothrix globosa, Sporothrix luriei, Sporothrix mexicana and S. schenckii. Cases of sporotrichosis have significantly increased in Brazil over the past decade, especially in the state of Rio de Janeiro (RJ), where an epidemic among cat owners has been observed. The zoonotic transmission from cats to humans suggests a common source of infection and indicates that animals can act as vectors. We performed a molecular characterisation of samples collected during the first outbreak of familial sporotrichosis caused by S. brasiliensis in the state of Espírito Santo, Brazil. These results represent the first description of such an outbreak outside the endemic area of zoonotic sporotrichosis in RJ

Comparison of different DNA-based methods for molecular typing of Histoplasma capsulatum

Submitted by Janaína Nascimento ([email protected]) on 2019-03-13T14:20:49Z No. of bitstreams: 1 ve_Muniz_Mauro_etal_INI_2010.pdf: 1298614 bytes, checksum: e9db0f1cbe38ec8d24467f8116ecda64 (MD5)Approved for entry into archive by Janaína Nascimento ([email protected]) on 2019-03-21T23:12:43Z (GMT) No. of bitstreams: 1 ve_Muniz_Mauro_etal_INI_2010.pdf: 1298614 bytes, checksum: e9db0f1cbe38ec8d24467f8116ecda64 (MD5)Made available in DSpace on 2019-03-21T23:12:43Z (GMT). No. of bitstreams: 1 ve_Muniz_Mauro_etal_INI_2010.pdf: 1298614 bytes, checksum: e9db0f1cbe38ec8d24467f8116ecda64 (MD5) Previous issue date: 2010Fundação Oswaldo Cruz. Instituto de Pesquisa Clínica Evandro Chagas. Laboratório de Micologia. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto de Pesquisa Clínica Evandro Chagas. Laboratório de Micologia. Rio de Janeiro, RJ, Brasil.Westmead Millennium Institute. Centre for Infectious Diseases and Microbiology. Molecular Mycology Research Laboratory. New South Wales, Australia / Westmead Hospital. New South Wales, Australia.Albert Einstein College of Medicine. Microbiology and Immunology. New York, NY, USA.Fundação Oswaldo Cruz. Instituto de Pesquisa Clínica Evandro Chagas. Laboratório de Micologia. Rio de Janeiro, RJ, Brasil.Histoplasma capsulatum is very prevalent in the environment and is one of the most common causes of mycoses in humans and diverse animals in Brazil. Multiple typing methods have been developed to study H. capsulatum epidemiology; however, there is limited information concerning comparisons of results obtained with different methods using the same set of isolates. To explore the diversity of H. capsulatum in Brazil and to determine correlations between the results of three different molecular typing techniques, we examined 51 environmental, animal, and human isolates by M13 PCR fingerprinting, PCR-restriction fragment length polymorphism (RFLP) analysis of the internal transcribed region 1 (ITS1)-5.8S-ITS2 region of the rDNA locus, and DNA sequencing and phylogenetic analysis of parts of four protein-encoding genes, the Arf (ADP ribosylation factor), H-anti (H antigen precursor), Ole (delta-9 fatty acid desaturase), and Tub1 (alpha-tubulin) genes. Each method identified three major genetic clusters, and there was a high level of concordance between the results of the typing techniques. The M13 PCR fingerprinting and PCR-RFLP analyses produced very similar results and separated the H. capsulatum isolates included in this study into three major groups. An additional approach used was comparison of our Brazilian ITS1-5.8S-ITS2 sequences with the sequences deposited previously in NCBI data banks. Our analyses suggest that H. capsulatum can be divided into different molecular types that are dispersed around the world. Our results indicate that the three methods used in this study are reliable and reproducible and that they have similar sensitivities. However, M13 PCR fingerprinting has some advantages over the other two methods as it is faster, cheaper, and more user friendly, which especially increases its utility for molecular typing of Histoplasma in situations where laboratory facilities are relatively limited

Evaluation of na Enzyme Linked Immunosorbent Assay (ELISA) using purified, deglycosylated histoplamin for different clinical manifestations of histoplamosis

Submitted by Repositório Arca ([email protected]) on 2019-04-24T16:34:41Z No. of bitstreams: 1 license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5)Approved for entry into archive by Janaína Nascimento ([email protected]) on 2019-12-10T13:52:40Z (GMT) No. of bitstreams: 2 va_Guimarães_Allan_etal_INI_2010.pdf: 499129 bytes, checksum: 9e8b3bb1bbcd366e32e8691d2c58f2a1 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5)Made available in DSpace on 2019-12-10T13:52:40Z (GMT). No. of bitstreams: 2 va_Guimarães_Allan_etal_INI_2010.pdf: 499129 bytes, checksum: 9e8b3bb1bbcd366e32e8691d2c58f2a1 (MD5) license.txt: 1748 bytes, checksum: 8a4605be74aa9ea9d79846c1fba20a33 (MD5) Previous issue date: 2010Yeshiva University. Albert Einstein College of Medicine. Department of Medicine & Microbiology and Immunology. Division of Infectious Diseases. New York, NY, USA.Fundação Oswaldo Cruz. Instituto de Pesquisa Clínica Evandro Chagas. Laboratório de Micologia. Setor de Imunodiagnóstico. Rio de Janeiro, RJ, Brasil.Fundação Oswaldo Cruz. Instituto de Pesquisa Clínica Evandro Chagas. Laboratório de Micologia. Setor de Imunodiagnóstico. Rio de Janeiro, RJ, Brasil.Universidade Federal do Rio de Janeiro. Instituto de Microbiologia Prof. Paulo de Góes. Rio de Janeiro, RJ, Brasil.Yeshiva University. Albert Einstein College of Medicine. Department of Medicine & Microbiology and Immunology. Division of Infectious Diseases. New York, NY, USA.Fundação Oswaldo Cruz. Instituto de Pesquisa Clínica Evandro Chagas. Laboratório de Micologia. Setor de Imunodiagnóstico. Rio de Janeiro, RJ, Brasil.Diagnosis of invasive fungal diseases remains problematic, especially in undeveloped countries. We have developed an enzyme-linked immunosorbent assay (ELISA) for the detection of antibodies to Histoplasma capsulatum using metaperiodate treated purified histoplasmin (ptHMIN). Our ELISA was validated comparing sera from patients with histoplasmosis, related mycoses, and healthy individuals. The overall test specificity was 96%, with sensitivities of 100% (8/8) in acute disease, 90% (9/10) in chronic disease, 89% (8/9) in disseminated infection in individuals without HIV infection, 86% (12/14) in disseminated disease in the setting of HIV infection and 100% (3/3) in mediastinal histoplasmosis. These parameters are superior to the use of untreated histoplasmin in diagnostic ELISAs. The high specificities, sensitivities, and simplicity of our ELISA support further development of a deglycosylated HMIN ELISA for clinical use and for monitoring the humoral immune response during therapy in patients with chronic and disseminated histoplasmosis

Could histoplasma capsulatum be related to healthcare-associated infections?

Healthcare-associated infections (HAI) are described in diverse settings. The main etiologic agents of HAI are bacteria (85%) and fungi (13%). Some factors increase the risk for HAI, particularly the use of medical devices; patients with severe cuts, wounds, and burns; stays in the intensive care unit, surgery, and hospital reconstruction works. Several fungal HAI are caused by Candida spp., usually from an endogenous source; however, cross-transmission via the hands of healthcare workers or contaminated devices can occur. Although other medically important fungi, such as Blastomyces dermatitidis, Paracoccidioides brasiliensis, and Histoplasma capsulatum, have never been considered nosocomial pathogens, there are some factors that point out the pros and cons for this possibility. Among these fungi, H. capsulatum infection has been linked to different medical devices and surgery implants. The filamentous form of H. capsulatum may be present in hospital settings, as this fungus adapts to different types of climates and has great dispersion ability. Although conventional pathogen identification techniques have never identified H. capsulatum in the hospital environment, molecular biology procedures could be useful in this setting. More research on H. capsulatum as a HAI etiologic agent is needed, since it causes a severe and often fatal disease in immunocompromised patients.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq