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Churro C.,Instituto Nacional Of Sade Dr Ricardo Jorge | Churro C.,University of Porto | Pereira P.,Instituto Nacional Of Sade Dr Ricardo Jorge | Vasconcelos V.,University of Porto
Archives of Microbiology | Year: 2012

A species-specific method to detect and quantify Planktothrix agardhii was developed by combining the SYBR Green I real-time polymerase chain reaction technique with a simplified DNA extraction procedure for standard curve preparation. Newly designed PCR primers were used to amplify a speciWc fragment within the rpoC1 gene. Since this gene exists in single copy in the genome, it allows the direct achievement of cell concentrations. The cell concentration determined by real-time PCR showed a linear correlation with the cell concentration determined from direct microscopic counts. The detection limit for cell quantification of the method was 8 cells μL-1, corresponding to 32 cells per reaction. Furthermore, the real-time qPCR method described in this study allowed a successful quantification of P. agardhii from environmental water samples, showing that this protocol is an accurate and economic tool for a rapid absolute quantification of the potentially toxic cyanobacterium P. agardhii. © Springer-Verlag 2012. Source


Caccio S.M.,Parasitic and Immunomediated Diseases | Beck R.,Parasitic and Immunomediated Diseases | Beck R.,Croatian Veterinary Institute | Almeida A.,Parasitic and Immunomediated Diseases | And 3 more authors.
Parasitology | Year: 2010

PCR assays have been developed mainly to assist investigations into the epidemiology of Giardia duodenalis, the only species in the Giardia genus having zoonotic potential. However, a reliable identification of all species is of practical importance, particularly when water samples and samples from wild animals are investigated. The aim of the present work was to genotype Giardia species and G. duodenalis assemblages using as a target the region spanning the 5.8S gene and the 2 flanking internal transcribed spacers (ITS1 and ITS2) of the ribosomal gene. Primers were designed to match strongly conserved regions in the 3 end of the small subunit and in the 5 end of the large subunit ribosomal genes. The corresponding region (about 310 bp) was amplified from 49 isolates of both human and animal origin, representing all G. duodenalis assemblages as well as G. muris and G. microti. Sequence comparison and phylogenetic analysis showed that G. ardeae, G. muris, G. microti as well as the 7 G. duodenalis assemblages can be easily distinguished. Since the major subgroups within the zoonotic assemblages A and B can be identified by sequence analysis, this assay is also informative for molecular epidemiological studies. © Cambridge University Press 2010. Source


Sabino R.,Instituto Nacional Of Sade Dr Ricardo Jorge | Sabino R.,University of Minho | Verissima C.,Instituto Nacional Of Sade Dr Ricardo Jorge | Brandao J.,Instituto Nacional Of Sade Dr Ricardo Jorge | And 8 more authors.
Medical Mycology | Year: 2010

This study presents data on the incidence of candidemia in a Portuguese oncology hospital during a 6-year period. The species distribution and their antifungal susceptibility, as well as the clinical outcomes associated with candidemia were evaluated. A total of 119 episodes were reported, with the majority occurring among patients older than 56 years. The most common underlying medical conditions were solid tumors (64.5%) and hematological disease (28.2%). The most frequent species found was Candida albicans (48.7%), followed by C. parapsilosis (20.2%), C. tropicalis (8.4%), C. krusei (6.7%) and C. glabrata (5.0%), but Saccharomyces cerevisiae and Rhodotorula mucilaginosa were also isolated. Candida albicans was more frequently associated with solid tumors of the gastrointestinal and genitourinary tracts and breast (P = 0.005), while non-C. albicans Candida species were most frequently recovered from hematological patients (P = 0.007). The mortality rate associated with candidemia was 31.9% (P = 0.016). All C. albicans and C. parapsilosis isolates were susceptible to fluconazole, voriconazole and itraconazole. Resistance to caspofungin was only observed in C. albicans and in the R. mucilaginosa isolates. Posaconazole was active against all C. parapsilosis isolates tested but resistant strains were found among C. albicans (4.9%), C. tropicalis (12.5%), C. krusei (25%) and C. glabrata (50%). This study provides useful information regarding the local epidemiology of candidemia in cancer patients. © 2010 ISHAM. Source


Pacheco S.A.,Instituto Nacional Of Sade Dr Ricardo Jorge | Aguiar F.,Instituto Nacional Of Sade Dr Ricardo Jorge | Ruivo P.,Instituto Do Desporto Of Portugal | Proenca M.C.,Instituto Nacional Of Sade Dr Ricardo Jorge | And 3 more authors.
Journal of Toxicology and Environmental Health - Part A: Current Issues | Year: 2012

Environmental tobacco smoke (ETS), also referred to as secondhand smoke (SHS), is a major threat to public health and is increasingly recognized as an occupational hazard to workers in the hospitality industry. Therefore, several countries have implemented smoke-free regulations at hospitality industry sites. In Portugal, since 2008, legislation partially banned smoking in restaurants and bars but until now no data have been made available on levels of indoor ETS pollution/exposure at these locations. The aim of this study was to examine the occupational exposure to ETS/SHS in several restaurants in Lisbon, measured by indoor fine particles (PM2.5) and urinary cotinine concentration in workers, after the partial smoking ban in Portugal. Results showed that the PM2.5 median level in smoking designated areas was 253 g/m 3, eightfold higher than levels recorded in canteens or outdoor. The nonsmoking rooms of mixed restaurants exhibited PM2.5 median level of 88 g/m3, which is higher than all smoke-free locations studied, approximately threefold greater than those found in canteens. Importantly, urinary cotinine concentrations were significantly higher in nonsmoker employees working in those smoking designated areas, confirming exposure to ETS. The proportion of smokers in those rooms was found to be significantly positively correlated with nonsmoker urinary cotinine and indoor PM2.5 levels, establishing that both markers were occupational-ETS derived. The use of reinforced ventilation systems seemed not to be sufficient to decrease the observed ETS pollution/exposure in those smoking locations. Taken together, these findings demonstrate that the partial restrictions on smoking in Portuguese venues failed to provide adequate protection to their employees, irrespective of protective measures used. Therefore, a smoke-free legislation protecting individuals from exposure to ETS/SHS in all public places and workplaces is urgently needed in Portugal. © 2012 Copyright Taylor and Francis Group, LLC. Source

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