Browsing by Author "Verissimo, Cristina"
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- Air fungal contamination in ten hospitals’ food units from LisbonPublication . Viegas, C.; Ramos, C.; Almeida, M.; Sabino, Raquel; Verissimo, Cristina; Rosado, L.A descriptive study was developed to monitor air fungal contamination in ten food units from hospitals. Fifty air samples of 250 litres were collected through impaction method. Samples were collected in food storage facilities, kitchen, food plating, canteen and also, outside premises, since this is the place regarded as reference. Simultaneously, environmental parameters were also monitored, including temperature and relative humidity through the equipment Babouc, LSI Sistems and according to the International Standard ISO 7726.
- Aspergilose pulmonar crónica: os desafios para o diagnóstico e aplicabilidade de ferramentas auxiliares - relato de casoPublication . Trápaga, Mariana Rodrigues; Martins Melo, Aryse; Poester, Vanice Rodrigues; Basso, Rossana Patricia; Sabino, Raquel; Verissimo, Cristina; Benelli, Jéssica Louise; Klafke, Gabriel Baracy; Orzechowski Xavier, MelissaIntrodução: Estima‐se que anualmente 3 milhões de pessoas em todo o mundo desenvolvam aspergilose pulmonar crônica (CPA), no entanto, seu diagnóstico é desafiador. Objetivo: Relatar o caso de um paciente com repetidos isolamentos de A. fumigatus em amostras respiratórias desde 2013, e diagnóstico de CPA confirmado somente em 2019. Metodologia: Homem, 73 anos, diagnóstico de HIV em 2000 (desde 2010 supressão virológica), ex‐usuário de drogas, tabaco e álcool (abstêmio desde 2008), timoma em 2002, com ressecção cirúrgica. Em 2006 e 2009, apresentou infecção por Mycobacterium avium. Desde 2010, em uso de corticóide inalatório e β2‐agonista para DPOC, infecções respiratórias de repetição e deterioração progressiva de parênquima pulmonar em exames de imagem. Realizadas cerca de 10 investigações para micobacteriose, após tratamento, todas negativas. Em 2013 e 2015, A. fumigatus foi isolado de escarro e LBA, respectivamente. Interpretado como colonização, não houve tratamento antifúngico em ambas ocasiões. Em 2018, A. fumigatus foi novamente isolado de escarro, sendo realizada investigação sorológica, que permitiu o diagnóstico por detecção de anticorpos (IDGA ‐ IMMY®; e ELISA IgG Aspergillus Bio‐Rad®) e antígeno (LFA Aspergillus GM, IMMY®). Paciente não tolerou a terapia com anfo B, recebendo itraconazol (ITC) (200 mg; 12/12 h). Após 6 meses de tratamento, teve melhora clínica e estabilização do quadro radiológico; e IDGA negativou, sendo indicada manutenção do ITC por mais 6 meses. A análise genotípica pela técnica de microssatélites (alto poder discriminatório: 0,9968), comprovou que três isolados de A. fumigatus obtidos em diferentes momentos eram a mesma estirpe. Discussão/Conclusão: O diagnóstico da CPA é um desafio pela dificuldade em interpretar o isolamento de A. fumigatus de amostra respiratória, podendo ser contaminação, colonização ou, de fato, uma infecção ativa. Nosso caso ilustra este contexto, no qual esse diagnóstico foi considerado somente após diversos isolamentos fúngicos. O fato de tratar‐se de mesma cepa fúngica isolada nos diferentes anos, sugere a associação deste agente com a deterioração progressiva do parênquima pulmonar; ou ainda uma colonização prévia que culminou com progressão para doença ativa após danos por outras etiologias e/ou uso de corticóide. Em ambos os casos, cabe ressaltar a importância de investigar um paciente com comprometimento pulmonar crônico cujas amostras respiratórias resultem em isolamento de A. fumigatus.
- Biofilms and catheter related bloodstream infection: a tale of two kigdomsPublication . Borges, Vítor; Wenner, Sigurd; Nogueira, Isabel; Faria, Isabel; Pessanha, Maria Ana; Verissimo, Cristina; Sabino, Raquel; Rodrigues, Joao; Matias, Rui; Martins, Filomena; Carvalho, Patricia; Gomes, Joao Paulo; Jordão, LuísaBackground: Biofilm-associated infections are a public health concern in the context of healthcare-associated infections (HAI) such as catheter-related bloodstream infections (CRBSI). Here, we studied two top ten CRBS etiological agents, Enterobacter cloacae and Candida parapsilosis, isolated from a patient with CRBSI in order to understand the role played by biofilms on this HAI. Materials/methods: E.cloacae and C.parapsilosis were isolated from CVC and peripheral blood by standard procedures. EUCAST guidelines were followed for antimicrobial susceptibility evaluation. Single and/or mixed biofilms were assembled on different materials in Mueller-Hinton broth with 2% glucose. Biofilm assembly was assessed by crystal violet assay and scanning electron microscopy (SEM). Fluorescence in situ hybridization (FISH) was used for identification and to assess microorganisms distribution within the biofilm (3D reconstruction). In addition, Focus Ion Beam (FIB)-SEM was used to assess biofilms assembled on inner and outer surfaces of CVCs and construct tomograms. CVC and hemoculture (HC) isolates were subjected to whole-genome sequencing (WGS). Results: All Enterobacter and Candida isolates were antimicrobial resistant. Of note, E. cloacae-CVC revealed an additional resistance (ceftolozame-tazobactam) in comparison to the HC- isolate. Both microorganisms assembled biofilms on glass, polystyrene and polyurethane. Mixed biofilms were denser when both microorganisms were present from the beginning. Biofilm phenotype was not dependent of biofilm initiation by E.cloacae or C.parapsilosis. FISH and SEM analysis showed that biofilm bottom layer was in all cases richer in E.cloacae. Environmental isolates of the same species were also tested, showing that this biofilm phenotype is not a general feature. Using polyurethane catheters (shape/material factor), we observed denser mixed biofilms richer in EPS. FIB-SEM preliminary results suggest that biofilms assembled on inner and outer catheter surface might differ on microorganisms’ distribution. WGS confirmed the genetic identity of the CVC/HC pairs while corroborating the virulence potential and antimicrobial resistant character of the CRBSI-driving pathogens. Conclusions: The results suggest that biofilms allow interaction and adaptation of microorganisms belonging to different kingdoms (Bacteria and Fungi). Adaptation might affect virulence in a transitory or permanent fashion, with potential impact on microorganisms’ potential to cause CRBSI.
- Diagnosis of Tinea pedis and onychomycosis in patients from Portuguese National Institute of Health Dr. Ricardo Jorge: a four year studyPublication . Viegas, Carla; Sabino, Raquel; Parada, Helena; Brandão, João; Carolino, Elisabete; Rosado, Laura; Verissimo, CristinaABSTRACT - Tinea pedis and onychomycosis are two rather diverse clinical manifestations of superficial fungal infections, and their etiologic agents may be dermatophytes, non-dermatophyte moulds or yeasts. This study was designed to statistically describe the data obtained as results of analysis conducted during a four year period on the frequency of Tinea pedis and onychomycosis and their etiologic agents. A questionnaire was distributed from 2006 to 2010 and answered by 186 patients, who were subjected to skin and/or nail sampling. Frequencies of the isolated fungal species were cross-linked with the data obtained with the questionnaire, seeking associations and predisposing factors. One hundred and sixty three fungal isolates were obtained, 24.2% of which composed by more than one fungal species. Most studies report the two pathologies as caused primarily by dermatophytes, followed by yeasts and lastly by non-dermatophytic moulds. Our study does not challenge this trend. We found a frequency of 15.6% of infections caused by dermatophytes (with a total of 42 isolates) of which T. rubrum was the most frequent species (41.4%). There was no significant association (p >0.05) among visible injury and the independent variables tested, namely age, gender, owning pet, education, swimming pools attendance, sports activity and clinical information. Unlike other studies, the variables considered did not show the expected influence on dermatomycosis of the lower limbs. It is hence necessary to conduct further studies to specifically identify which variables do in fact influence such infections.
- Diagnóstico laboratorial das infeções fúngicas invasivasPublication . Verissimo, CristinaAs infeções fúngicas invasivas (IFI) estão associadas a elevada mortalidade. O diagnóstico numa fase inicial é determinante para o sucesso da terapia. As leveduras do género Candida são os agentes mais frequentes de infeção fúngica invasiva; No grupo dos fungos filamentosos o género Aspergillus e os fungos asseptados da ordem Mucorales são referidos como os principais agentes de IFI, no entanto outras espécies têm vindo a emergir, nomedamente Rhodotorula spp. Trichopoporon spp., Fusarium spp. Scedosporium spp. Devido à falta de métodos capazes de diagnosticar com exatidão as infeções fungicas invasivas o diagnóstico destas infeções deverá ser baseado em várias abordagens complementares que incluem: critérios clínicos, imagiologia compatível, histopatologia, cultura e métodos não culturais (pesquisa de antigénio, DNA fúngico). Os métodos moleculares têm vindo a ser melhorados de modo a assegurar reprodutibilidade intra e inter laboratorial, com vista à sua validação para diagnóstico laboratorial. Perspetivam-se novas metodologias que permitam tornar mais rápido o diagnóstico das infeções fúngicas invasivas e guidelines incluam os métodos não culturais, com vista a melhor adequação da terapia antifúngica. Nesta apresenção serão abordadas as diferentes metodologias disponiveis para o diagnóstico laboratorial das infeções fúngicas, principais vantagens e limitações.
- Diagnóstico laboratorial em micologiaPublication . Verissimo, CristinaObjetivos: 1.Principais tipos de infecção; 2.Colheita e transporte de amostras; 3.Métodos diretos (exame direto e cultural); 4.Métodos indiretos para diagnóstico de infeções fúngicas; 5.Valorização de resultados; 6.Identificação de leveduras e fungos filamentosos.
- Fungal Agents agents of cutaneous and subcutaneous fungal infections - 5 years experiencePublication . Verissimo, Cristina; Simões, Helena; Sabino, RaquelIntroduction: Fungal infections involving subcutaneous tissue often develop following a penetrating wound through the skin. The etiological agents are usually soil fungi or decomposers of plant material decomposers. Fungal infections involving subcutaneous tissue are frequently grouped as (I) mycetoma (II) chromoblastomycosis which the etiologic agent develops dematiaceous muriform cells in the tissue (III) subcutaneous phaehyphomycosis caused by dematiaceous fungi and (IV) hyalohyphomycosis caused by non dematiaceoushyalin fungi. Mycetoma and chromoblastomycosis are common in subtropical and tropical regions of the world but rare in Europe where subcutaneous infections have a different epidemiology. This work aims to evaluate the epidemiology of cutaneous/subcutaneous infections diagnosed at in our laboratory during a 5 year period.
- Fungal contamination of poultry litter: a public health problemPublication . Viegas, Carla; Carolino, Elisabete; Malta-Vacas, Joana; Sabino, Raquel; Viegas, Susana; Verissimo, CristinaAlthough numerous studies have been conducted on microbial contaminants associated with various stages related to poultry and meat products processing, only a few reported on fungal contamination of poultry litter. The goals of this study were to (1) characterize litter fungal contamination and (2) report the incidence of keratinophilic and toxigenic fungi presence. Seven fresh and 14 aged litter samples were collected from 7 poultry farms. In addition, 27 air samples of 25 litters were also collected through impaction method, and after laboratory processing and incubation of collected samples, quantitative colony-forming units (CFU/m3) and qualitative results were obtained. Twelve different fungal species were detected in fresh litter and Penicillium was the most frequent genus found (59.9%), followed by Alternaria (17.8%), Cladosporium (7.1%), and Aspergillus (5.7%). With respect to aged litter, 19 different fungal species were detected, with Penicillium sp. the most frequently isolated (42.3%), followed by Scopulariopsis sp. (38.3%), Trichosporon sp. (8.8%), and Aspergillus sp. (5.5%). A significant positive correlation was found between litter fungal contamination (CFU/g) and air fungal contamination (CFU/m3). Litter fungal quantification and species identification have important implications in the evaluation of potential adverse health risks to exposed workers and animals. Spreading of poultry litter in agricultural fields is a potential public health concern, since keratinophilic (Scopulariopsis and Fusarium genus) as well as toxigenic fungi (Aspergillus, Fusarium, and Penicillium genus) were isolated.
- Fungal Contamination of Sandpits from Recreational Parks and Schools: a Potential Risk for Human HealthPublication . Viegas, Carla; Brandão, João; Sabino, Raquel; Meneses, Marcia; Verissimo, CristinaSandpits used by children are frequently visited by wild life which constitutes a source of fungal pathogens and allergenic fungi. This study aimed to take an unannounced snapshot of the urban levels of fungal contaminants in sands, using for this purpose two public recreational parks, three elementary schools and two kindergartens. All samples were from Lisbon and neighboring municipalities and were tested for fungi of clinical interest. Potentially pathogenic fungi were isolated from all samples besides one. Fusarium dimerum (32.4%) was found to be the dominant species in one park and Chrysonilia spp. in the other (46.6%). Fourteen different species and genera were detected and no dermatophytes were found. Of a total of 14 species and genera, the fungi most isolated from the samples of the elementary schools were Penicillium spp. (74%), Cladophialophora spp. (38%) and Cladosporium spp. (90%). Five dominant species and genera were isolated from the kindergartens. Penicillium spp. was the only genus isolated in one, though with remarkably high counts (32500 colony forming units per gram). In the other kindergarten Penicillium spp. were also the most abundant species, occupying 69% of all the fungi found. All of the samples exceeded the Maximum Recommended Value (MRV) for beach sand defined by Brandão et al. 2011, which are currently the only quantitative guidelines available for the same matrix. The fungi found confirm the potential risk of exposure of children to keratinophilic fungi and demonstrates that regular cleaning or replacing of sand needs to be implemented in order to minimize contamination.
- Fungos e ambiente hospitalar: preocupações relevantes e riscos para o doentePublication . Sabino, Raquel; Francisco, M.; Viegas, C.; Martins, C.; Verissimo, CristinaO número de infeções fúngicas nosocomiais tem vindo a aumentar drasticamente sendo causa, cada vez mais comum, de elevada morbilidade e mortalidade em doentes hospitalizados. O grau de contaminação fúngica ambiental é considerado um fator de extrema importância na incidência e epidemiologia destas infeções. Em contexto hospitalar, a infeção fúngica poderá ocorrer por contacto direto ou indireto com superfícies ou objetos contaminados, através das mãos de profissionais de saúde e por ingestão ou inalação de partículas ou bioaerossóis contaminados. As espécies de Aspergillus e Candida são responsáveis pela maioria das infeções fúngicas em pacientes imunocomprometidos e são frequentemente isolados em ambiente hospitalar. Descrevem-se neste trabalho dois case-study em Hospitais da região de Lisboa onde para além da caracterização do ambiente hospitalar quanto às espécies fúngicas predominantes, foram efetuadas pesquisas dirigidas aos géneros Aspergillus e Candida. Verificou-se que o controlo das infeções nosocomiais requer um conhecimento aprofundado da espécie/estirpe infetante, bem como do seu padrão de suscetibilidade aos antifúngicos, determinando assim o risco de infeção tanto para doentes como para os profissionais de saúde.
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