Accessing indoor fungal contamination using conventional and molecular methods in Portuguese poultries

Epidemiological studies showed increased prevalence of respiratory symptoms and adverse changes in pulmonary function parameters in poultry workers, corroborating the increased exposure to risk factors, such as fungal load and their metabolites. This study aimed to determine the occupational exposure threat due to fungal contamination caused by the toxigenic isolates belonging to the complex of the species of Aspergillus flavus and also isolates fromAspergillus fumigatus species complex. The study was carried out in seven Portuguese poultries, using cultural and molecularmethodologies. For conventional/cultural methods, air, surfaces, and litter samples were collected by impaction method using the Millipore Air Sampler. For the molecular analysis, air samples were collected by impinger method using the Coriolis μ air sampler. After DNA extraction, samples were analyzed by real-time PCR using specific primers and probes for toxigenic strains of the Aspergillus flavus complex and for detection of isolates from Aspergillus fumigatus complex. Through conventional methods, and among the Aspergillus genus, different prevalences were detected regarding the presence of Aspergillus flavus and Aspergillus fumigatus species complexes, namely: 74.5 versus 1.0% in the air samples, 24.0 versus 16.0% in the surfaces, 0 versus 32.6% in new litter, and 9.9 versus 15.9%in used litter. Through molecular biology, we were able to detect the presence of aflatoxigenic strains in pavilions in which Aspergillus flavus did not grow in culture. Aspergillus fumigatus was only found in one indoor air sample by conventional methods. Using molecular methodologies, however, Aspergillus fumigatus complex was detected in seven indoor samples from three different poultry units. The characterization of fungal contamination caused by Aspergillus flavus and Aspergillus fumigatus raises the concern of occupational threat not only due to the detected fungal load but also because of the toxigenic potential of these species.

O uso de palmilhas de contato total e o impacte nas alterações posturais no trabalho em ortostatismo

Mestrado em Higiene e Segurança no Trabalho

Occupational exposure to toxigenic fungi from Aspergillus flavus complex

Bioaerosols are mainly composed of fungal particles, bacteria and plant spores, being fungi responsible for the release of VOCs and micotoxins into indoor environments. Aspergillus flavus is a common opportunistic pathogen causing human infections and is involved in the production of aflatoxin and other secondary metabolites associated with toxic and allergic reactions. Poultry workers are exposed to high concentrations of fungi and are therefore more prone to develop associated pathologies. To evaluate occupational exposure of the workers to Aspergillus flavus and aflatoxins, six animal production facilities were selected, including 10 buildings, from which indoor air samples and outdoor reference samples were obtained. Twenty-five duplicate samples were collected by two methodologies: impactation onto malt extract agar of 25L air samples using a Millipore Air Tester were used to evaluate quantitative (CFU/m3) and qualitative (species identification, whenever possible) sample composition; 300 L air samples collected with the Coriolis Air Sampler into phosphate–saline buffer were used to isolate DNA, following molecular identification of Aspergillus section flavi using nor-1 specific primers by real-time PCR.

Comparison of Aspergillus species-complexes detected in different environmental settings

Purpose: Samples from different environmental sources were screened for the presence of Aspergillus, and the distribution of the different species-complexes was determined in order to understand differences among that distribution in the several environmental sources and which of these species complexes are present in specific environmental settings. Methods: Four distinct environments (beaches, poultries, swineries and hospital) were studied and analyzed for which Aspergillus complexes were present in each setting. After plate incubation and colony isolation, morphological identification was done using macro- and microscopic characteristics. The universal fungal primers ITS1 and ITS4 were used to amplify DNA from all Aspergillus isolates, which was sequenced for identification to species complex level. SPSS v15.0 for Windows was used to perform the statistical analysis. Results: Thirty-nine isolates of Aspergillus were recovered from both the sand beach and poultries, 31 isolates from swineries, and 80 isolates from hospital environments, for a total 189 isolates. Eleven species complexes were found total. Isolates belonging to the Aspergillus Versicolores species-complex were the most frequently found (23.8%), followed by Flavi (18.0%), Fumigati (15.3%) and Nigri (13.2%) complexes. A significant association was found between the different environmental sources and the distribution of the several species-complexes (p<0.001); the hospital environment had a greater variability of species-complexes than other environmental locations (10 in hospital environment, against nine in swine, eight in poultries and seven in sand beach). Isolates belonging to Nidulantes complex were detected only in the hospital environment, whereas the other complexes were identified in more than one setting. Conclusion: Because different Aspergillus complexes have different susceptibilities to antifungal drugs, and different abilities in producing mycotoxins, knowledge of the species-complex epidemiology for each setting may allow preventive or corrective measures to be taken toward decreasing professional workers or patient exposure to those agents.

Estratégias e internacionalização na manutenção e produção

Trabalho Final de Mestrado para obtenção do grau de Mestre em Engenharia Mecânica

Molecular screening of 246 Portuguese Aspergillus isolates among different clinical and environmental sources

Clinical and environmental samples from Portugal were screened for the presence of Aspergillus and the distributions of the species complexes were determined in order to understand how their distributions differ based on their source. Fifty-seven Aspergillus isolates from clinical samples were collected from 10 health institutions. Six species complexes were detected by internal transcribed spacer sequencing; Fumigati, Flavi, and Nigri were found most frequently (50.9%, 21.0%, and 15.8%, respectively). β-tubulin and calmodulin sequencing resulted in seven cryptic species (A. awamorii, A. brasiliensis, A. fructus, A. lentulus, A. sydowii, A. tubingensis, Emericella echinulata) being identified among the 57 isolates. Thirty-nine isolates of Aspergillus were recovered from beach sand and poultry farms, 31 from swine farms, and 80 from hospital environments, for a total 189 isolates. Eleven species complexes were found in these 189 isolates, and those belonging to the Versicolores species complex were found most frequently (23.8%). There was a significant association between the different environmental sources and distribution of the species complexes; the hospital environment had greater variability of species complexes than other environmental locations. A high prevalence of cryptic species within the Circumdati complex was detected in several environments; from the isolates analyzed, at least four cryptic species were identified, most of them growing at 37ºC. Because Aspergillus species complexes have different susceptibilities to antifungals, knowing the species-complex epidemiology for each setting, as well as the identification of cryptic species among the collected clinical isolates, is important. This may allow preventive and corrective measures to be taken, which may result in decreased exposure to those organisms and a better prognosis.

Morfometria do trato vocal por ressonância magnética: simulação de padrões patológicos articulatórios

Introdução – A análise da forma ou morfometria de estruturas anatómicas, como o trato vocal, pode ser efetuada a partir de imagens bidimensionais (2D) como de aquisições volumétricas (3D) de ressonância magnética (RM). Esta técnica de imagem tem vindo a ter uma utilização crescente no estudo da produção da fala. Objetivos – Demonstrar como pode ser efetuada a morfometria do trato vocal a partir da imagem por ressonância magnética e ainda apresentar padrões anatómicos normais durante a produção das vogais [i a u] e dois padrões articulatórios patológicos em contexto simulado. Métodos – As imagens consideradas foram recolhidas a partir de aquisições 2D (Turbo Spin-eco) e 3D (Flash Gradiente-Eco) de RM em quatro sujeitos durante a produção das vogais em estudo; adicionalmente procedeu-se à avaliação de duas perturbações articulatórias usando o mesmo protocolo de RM. A morfometria do trato vocal foi extraída com recurso a técnicas manuais (para extração de cinco medidas articulatórias) e automáticas (para determinação de volumes) de processamento e análise de imagem. Resultados – Foi possível analisar todo o trato vocal, incluindo a posição e a forma dos articuladores, tendo por base cinco medidas descritivas do posicionamento destes órgãos durante a produção das vogais. A determinação destas medições permitiu identificar quais as estratégias mais comummente adotadas na produção de cada som, nomeadamente a postura articulatória e a variação de cada medida para cada um dos sujeitos em estudo. No contexto de voz falada intersujeitos, foi notória a variabilidade nos volumes estimados do trato vocal para cada som e, em especial, o aumento do volume do trato vocal na perturbação articulatória de sigmatismo. Conclusão – A imagem por RM é, sem dúvida, uma técnica promissora no estudo da fala, inócua, não-invasiva e que fornece informação fiável da morfometria do trato vocal.

Mammography equipment design: impact on radiographers’ practice

Objectives - Identify radiographers’ postures during frequent mammography procedures related to the mammography equipment and patient characteristics. Methods - A postural task analysis was performed using images acquired during the simulation of mammography positioning procedures. Simulations included craniocaudal/(CC) and mediolateral-oblique/(MLO) positioning in three different settings: radiographers and patients with similar statures, radiographers smaller than the patients and radiographers taller than the patients. Measurements of postural angles were performed by two raters using adequate software and classified according to the European Standard EN1005-4:2005 + A1:2008. Results - The simulations revealed that the most awkward posture in mammography is during the positioning of MLO projection in short-stature patients. Postures identified as causing work-related musculoskeletal disorder (WRMSD) risk were neck extension, arms elevated and the back stooped, presenting angles of 87.2, 118.6 and 63.6, respectively. If radiographers were taller than patients, then the trunk and arm postures were not acceptable. Conclusions - Working in a mammography room leads to awkward postures that can have an impact on radiographers’ health, namely WRMSDs. The results in this study showed that there are non-acceptable postures associated with frequent working procedures in mammography. MLO is the most demanding procedure for radiographer postures and may be related to WRMSDs. Mammography devices should be redesigned considering adjustability for radiographers. Main Messages: • Mammography constraints for radiographers in mammography procedures have not been well studied. • Performing mammography leads to awkward postures that can impact radiographers’ health. • MLO, the most demanding procedure for radiographers, is possibly related to WRMSDs.

Assessment of workers’ exposure to Aflatoxin B1 in a Portuguese waste industry

Aflatoxin B1 (AFB1) is considered by different International Agencies as a genotoxic and potent hepatocarcinogen. However, despite the fact that the fungi producing this compound are detected in some work environments, AFB1 is rarely monitored in occupational settings. The aim of the present investigation was to assess exposure to AFB1 of workers from one Portuguese waste company located in the outskirt of Lisbon. Occupational exposure assessment to AFB1 was done with a biomarker of internal dose that measures AFB1 in the serum by enzyme-linked immunosorbent assay. Forty-one workers from the waste company were enrolled in this study (26 from sorting; 9 from composting; 6 from incineration). A control group (n = 30) was also considered in order to know the AFB1 background levels for the Portuguese population. All the workers showed detectable levels of AFB1 with values ranging from 2.5ng ml−1 to 25.9ng ml−1 with a median value of 9.9±5.4ng ml−1. All of the controls showed values below the method’s detection limit. Results obtained showed much higher (8-fold higher) values when compared with other Portuguese settings already studied, such as poultry and swine production. Besides this mycotoxin, other mycotoxins are probably present in this occupational setting and this aspect should be taken into consideration for the risk assessment process due to possible synergistic reactions. The data obtained suggests that exposure to AFB1 occurs in a waste management setting and claims attention for the need of appliance of preventive and protective safety measures.

Posturas ensinadas em Portugal no início da aprendizagem de contrabaixo: relatório de estágio

Resumo I – Prática Pedagógica. No início da aula de contrabaixo pergunta-se: «queres tocar em pé ou sentado?» É uma pergunta que poucos professores fazem, não só porque exige que o aluno saiba tocar das duas maneiras, mas principalmente porque cada professor tem a sua escola e opta por uma forma de ensinar, que acha que é a melhor para o aluno. Será mais conveniente dar a escolher ou determinar a posição com que o aluno vai tocar? Neste trabalho, procuram-se as respostas, analisando as várias posturas que se podem ensinar quando se toca contrabaixo. Começando pela questão elementar de estar sentado ou em pé e passando pelos vários detalhes que determinam o posicionamento do aluno em relação ao instrumento, como por exemplo, a altura a que se põe o instrumento ou a forma de apoio dos pés, acaba-se por fazer uma reflexão sobre a melhor forma de ensinar nos dias de hoje. A prática pedagógica deste ano letivo de estágio, consiste concretamente na observação das posturas adotadas por três alunos em particular. Foram ensinadas novas posturas a dois alunos e aperfeiçoadas as posturas já ensinadas a uma terceira aluna. Os resultados finais foram reveladores para a investigação que foi feita em simultâneo.

Mediação de Conflitos Interpessoais

Relatório da UC Seminário de Apoio à Prática Profissional Supervisionada Mestrado em Educação Pré-Escolar

O brincador: a interação adulto-criança no momento da brincadeira

Relatório da UC Prática Profissional Supervisionada Mestrado em Educação Pré-Escolar

Efeitos da sobrecarga das mochilas escolares nas alterações posturais e de equilíbrio

Mestrado em Fisioterapia

Contributo para a validação de uma escala pediátrica, Peabody Developmental Motor Scale-2 (versão portuguesa)

Mestrado em Fisioterapia

O contributo da rotina Ler, contar e mostrar para o desenvolvimento de competências da leitura e de expressão oral

Relatório de estágio apresentado à Escola Superior de Educação de Lisboa para obtenção de grau de mestre em Ensino do 1º e 2º ciclo do Ensino Básico