Development of Detection Systems for the Sporangia of Peronospora Destructor

A monoclonal antibody that recognises components of the wall of sporangia of Peronospora destructor was raised. Tests using spores of higher fungi and other species of mildew demonstrated the specificity of the monoclonal. The antibody was used to develop lateral flow devices for sporangia of P. destructor. A competitive lateral flow format was developed which could detect onion downy mildew sporangia. Five-microliter gold anti-mouse IgM solution pre-mixed with 10 μl of P. destructor monoclonal antibody (EMA 242) proved the optimal concentration for detection of sporangia of P. destructor when applied to sample pads of lateral flow devices. Limits of approximately 500 sporangia of P. destructor could be detected by the absence of a test line on the lateral flow device within test samples. Using a scanning densitometer improved the sensitivity of detection. Further development and validation of the test is required if it is to be used for risk assessments of onion downy mildew in the field.

Determination of Alternaria spp. Habitats Using 7-Day Volumetric Spore Trap, Hybrid Single Particle Lagrangian Integrated Trajectory Model and Geographic Information System

There are many species among the Alternaria genus, which hosts on economically important crops causing significant yield losses. Less attention has been paid to fungi hosting on plants constituting substantial components of pastures and meadows. Alternaria spp. spores are also recognised as important allergens. A 7-day volumetric spore trap was used to monitor the concentration of airborne fungal spores. Air samples were collected in Worcester, England (2006–2010). Days with a high spore count were then selected. The longest episode that occurred within a five year study was chosen for modelling. Two source maps presenting distribution of crops under rotation and pastures in the UK were produced. Back trajectories were calculated using the HYSPLIT model. In ArcGIS clusters of trajectories were studied in connection with source maps by including the height above ground level and the speed of the air masses. During the episode no evidence for a long distance transport from the continent of Alternaria spp. spores was detected. The overall direction of the air masses fell within the range from South-West to North. The back trajectories indicated that the most important sources of Alternaria spp. spores were located in the West Midlands of England.

Application of Redundancy Analysis for Aerobiological Data

An aerobiological survey was conducted through five consecutive years (2006–2010) at Worcester (England).The concentration of 20 allergenic fungal spore types was measured using a 7-day volumetric spore trap. The relationship between investigated fungal spore genera and selected meteorological parameters (maximum, minimum, mean and dew point temperatures, rainfall, relative humidity, air pressure,wind direction) was examined using an ordination method(redundancy analysis) to determine which environmental factors favoured their most abundance in the air and whether it would be possible to detect similarities between different genera in their distribution pattern. Redundancy analysis provided additional information about the biology of the studied fungi through the results of the Spearman’s rank correlation. Application of the variance inflation factor in canonical correspondence analysis indicated which explanatory variables were auto-correlated and needed to be excluded from further analyses. Obtained information will be consequently implemented in the selection of factors that will be a foundation for forecasting models for allergenic fungal spores in the future.

Potential Sources of Airborne Alternaria spp. spores in South-west Spain

Fungi belonging to the genus of Alternaria are recognised as being significant plant pathogens, and Alternaria allergens are one of themost important causes of respiratory allergic diseases in Europe. This study aims to provide a detailed and original analysis of Alternaria transport dynamics in Badajoz, SW Spain. This was achieved by examining daily mean and hourly observations of airborne Alternaria spores recorded during days with high airborne concentrations of Alternaria spores (N100 s m−3) from 2009 to 2011, as well as four inventory maps of major Alternaria habitats, the overall synoptic weather situation and analysis of air mass transport using Hybrid Single Particle Lagrangian Integrated Trajectory model and geographic information systems. Land use calculated within a radius of 100 km from Badajoz shows that crops and grasslands are potentially the most important local sources of airborne Alternaria spores recorded at the site. The results of back trajectory analysis showthat, during the examined four episodes, the two main directions where Alternaria source areas were located were: (1) SW–W; and (2) NW–NE. Regional scale and long distance transport could therefore supplement the airborne catch recorded at Badajoz with Alternaria conidia originating from sources such as crops and orchards situated in other parts of the Iberian Peninsula.

Differing methodologies to measure microbe dispersal during hand drying

Background: Having previously investigated the dispersal by different hand drying methods of a chemical indicator, fungi and bacteria on the hands of users, this new study assessed the potential for viral dispersal. Aims/Objectives: To determine differences between hand drying methods in their capacity to disperse viruses on the hands of users to other occupants of public washrooms and into the washroom environment. Method: A harmless virus was used to artificially contaminate the hands of participants prior to using three different hand drying devices (jet air dryer, warm air dryer, paper towel dispenser). Viral dispersal was assessed at different heights and distances from the hand drying devices and also at different times after use by means of an air sampler. Results: The jet air dryer was shown to produce significantly more dispersal of virus than the warm air dryer or paper towels. After use of the jet air dryer, high numbers of virus were detected at a range of heights with maximum numbers between 0.61 and 1.22 metres. Virus was also detected at distances of up to 3 metres from the jet air dryer and in the air for up to 15 minutes after its use. The warm air dryer and paper towel dispenser produced low or zero viral counts at different heights, different distances and times after use. Conclusion: Jet air dryers have a greater potential than other hand drying methods to disperse viruses on the hands and contaminate other occupants of a public washroom and the washroom environment.

The risk of viral dispersal and aerosolization by different hand-drying methods

Background World Health Organization hand hygiene guidelines state that if electric hand dryers are used, they should not aerosolize pathogens. Previous studies have investigated the dispersal by different hand-drying devices of chemical indicators, fungi and bacteria on the hands. This study assessed the aerosolization and dispersal of virus on the hands to determine any differences between hand-drying devices in their potential to contaminate other occupants of public washrooms and the washroom environment. Methods A suspension of MS2, an Escherichia coli bacteriophage virus, was used to artificially contaminate the hands of participants prior to using three different handdrying devices: jet air dryer, warm air dryer, paper towel dispenser. Virus was detected by plaque formation on agar plates layered with the host bacterium. Vertical dispersal of virus was assessed at a fixed distance (0.4 m) and over a range of different heights (0.0 – 1.8 m) from the floor. Horizontal dispersal was assessed at different distances of up to three metres from the hand-drying devices. Virus aerosolization and dispersal was also assessed at different times up to 15 minutes after use by means of air sampling at two distances (0.1 and 1.0 m) and at a distance behind and offset from each of the hand-drying devices. Results Over a range of heights, the jet air dryer was shown to produce over 60 times greater vertical dispersal of virus from the hands than a warm air dryer and over 1300 times greater than paper towels; the maximum being detected between 0.6 and 1.2 metres from the floor. Horizontal dispersal of virus by the jet air dryer was over 20 times greater than a warm air dryer and over 190 times greater than paper towels; virus being detected at distances of up to three metres. Air sampling at three different positions from the hand-drying devices 15 minutes after use showed that the jet air dryer produced over 50-times greater viral contamination of the air than a warm air dryer and over 110-times greater than paper towels. Conclusions Due to their high air speed, jet air dryers aerosolize and disperse more virus over a range of heights, greater distances, and for longer times than other hand drying devices. If hands are inadequately washed, they have a greater potential to contaminate other occupants of a public washroom and the washroom environment. Main messages: Jet air dryers with claimed air speeds of over 600 kph have a greater potential than warm air dryers or paper towels to aerosolize and disperse viruses on the hands of users. The choice of hand-drying device should be carefully considered. Jet air dryers may increase the risk of transmission of human viruses, such as norovirus, particularly if hand washing is inadequate.

Sex-specific disruption of murine midbrain astrocytic and dopaminergic developmental trajectories following antenatal GC treatment

The mammalian midbrain dopaminergic systems arising in the substantia nigra pars compacta (SNc) and ventral tegmental area (VTA) are critical for coping behaviours and are implicated in neuropsychiatric disorders where early life challenges comprise significant risk factors. Here, we aimed to advance our hypothesis that glucocorticoids (GCs), recognised key players in neurobiological programming, target development within these systems, with a novel focus on the astrocytic population. Mice received antenatal GC treatment (AGT) by including the synthetic GC, dexamethasone, in the mothers' drinking water on gestational days 16-19; controls received normal drinking water. Analyses of regional shapes and volumes of the adult SNc and VTA demonstrated that AGT induced long-term, dose-dependent, structural changes that were accompanied by profound effects on astrocytes (doubling/tripling of numbers and/or density). Additionally, AGT induced long-term changes in the population size and distribution of SNc/VTA dopaminergic neurons, confirming and extending our previous observations made in rats. Furthermore, glial/neuronal structural remodelling was sexually dimorphic and depended on the AGT dose and sub-region of the SNc/VTA. Investigations within the neonatal brain revealed that these long-term organisational effects of AGT depend, at least in part, on targeting perinatal processes that determine astrocyte density and programmed cell death in dopaminergic neurons. Collectively, our characterisation of enduring, AGT-induced, sex-specific cytoarchitectural disturbances suggests novel mechanistic links for the strong association between early environmental challenge (inappropriate exposure to excess GCs) and vulnerability to developing aberrant behaviours in later life, with translational implications for dopamine-associated disorders (such as schizophrenia, ADHD, autism, depression), which typically show a sex bias

Comparison of virus dispersal and aerosolization by different hand-drying devices

Background World Health Organization and EU hand hygiene guidelines state that if electric hand dryers are used, they should not aerosolize pathogens. Previous studies have investigated the dispersal by different hand-drying devices of chemical indicators, fungi and bacteria on the hands. This study assessed the aerosolization and dispersal of virus on the hands to determine any differences between hand-drying devices in their potential to contaminate other occupants of public washrooms and the washroom environment. Methods A suspension of MS2, an Escherichia coli bacteriophage virus, was used to artificially contaminate the hands of participants prior to using three different hand-drying devices: jet air dryer, warm air dryer, paper towel dispenser. Virus was detected by plaque formation on agar plates layered with the host bacterium. Vertical dispersal of virus was assessed at a fixed distance (0.4 m) and over a range of different heights (0.0 – 1.8 m) from the floor. Horizontal dispersal was assessed at different distances of up to three metres from the hand-drying devices. Virus aerosolization and dispersal was also assessed at different times up to 15 minutes after use by means of air sampling at two distances (0.1 and 1.0 m) and at a distance behind and offset from each of the hand-drying devices. Results Over a range of heights, the jet air dryer was shown to produce over 60 times greater vertical dispersal of virus from the hands than a warm air dryer and over 1300 times greater than paper towels; the maximum being detected between 0.6 and 1.2 metres from the floor. Horizontal dispersal of virus by the jet air dryer was over 20 times greater than a warm air dryer and over 190 times greater than paper towels; virus being detected at distances of up to three metres. Air sampling at three different positions from the hand-drying devices 15 minutes after use showed that the jet air dryer produced over 50-times greater viral contamination of the air than a warm air dryer and over 110-times greater than paper towels. Conclusions Due to their high air speed, jet air dryers aerosolize and disperse more virus over a range of heights, greater distances, and for longer times than other hand drying devices. If hands are inadequately washed, they have a greater potential to contaminate other occupants of a public washroom and the washroom environment. Main messages: Jet air dryers with claimed air speeds of over 600 kph have a greater potential than warm air dryers or paper towels to aerosolize and disperse viruses on the hands of users. The choice of hand-drying device should be carefully considered. Jet air dryers may increase the risk of transmission of human viruses, such as norovirus, particularly if hand washing is inadequate.

Morte de enxertos de castanheira do Pará.

1972

Avaliação, tratamento e consolidação de madeira em edifícios antigos

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

Fungal contamination in two Portuguese wastewater treatment plants

The presence of filamentous fungi was detected in wastewater and air collected at wastewater treatment plants (WWTP) from several European countries. The aim of the present study was to assess fungal contamination in two WWTP operating in Lisbon. In addition, particulate matter (PM) contamination data was analyzed. To apply conventional methods, air samples from the two plants were collected through impaction using an air sampler with a velocity air rate of 140 L/min. Surfaces samples were collected by swabbing the surfaces of the same indoor sites. All collected samples were incubated at 27°C for 5 to 7 d. After lab processing and incubation of collected samples, quantitative and qualitative results were obtained with identification of the isolated fungal species. For molecular methods, air samples of 250 L were also collected using the impinger method at 300 L/min airflow rate. Samples were collected into 10 ml sterile phosphate-buffered saline with 0.05% Triton X-100, and the collection liquid was subsequently used for DNA extraction. Molecular identification of Aspergillus fumigatus and Stachybotrys chartarum was achieved by real-time polymerase chain reaction (RT-PCR) using the Rotor-Gene 6000 qPCR Detection System (Corbett). Assessment of PM was also conducted with portable direct-reading equipment (Lighthouse, model 3016 IAQ). Particles concentration measurement was performed at five different sizes: PM0.5, PM1, PM2.5, PM5, and PM10. Sixteen different fungal species were detected in indoor air in a total of 5400 isolates in both plants. Penicillium sp. was the most frequently isolated fungal genus (58.9%), followed by Aspergillus sp. (21.2%) and Acremonium sp. (8.2%), in the total underground area. In a partially underground plant, Penicillium sp. (39.5%) was also the most frequently isolated, also followed by Aspergillus sp. (38.7%) and Acremonium sp. (9.7%). Using RT-PCR, only A. fumigatus was detected in air samples collected, and only from partial underground plant. Stachybotrys chartarum was not detected in any of the samples analyzed. The distribution of particle sizes showed the same tendency in both plants; however, the partially underground plant presented higher levels of contamination, except for PM2.5. Fungal contamination assessment is crucial to evaluating the potential health risks to exposed workers in these settings. In order to achieve an evaluation of potential health risks to exposed workers, it is essential to combine conventional and molecular methods for fungal detection. Protective measures to minimize worker exposure to fungi need to be adopted since wastewater is the predominant internal fungal source in this setting.

Diagnóstico de estruturas de madeira em serviço num palacete do século XIX

Dissertação para obtenção do grau de Mestre em Engenharia Civil na Área de Especialização em Edificações

Qualidade do ar interior em Escolas Básicas de 1.º ciclo: QAI em ambientes escolares: estudo orientado para as Escolas Básicas de 1.º ciclo de um agrupamento escolar

A Qualidade do Ar Interior (QAI) é um fator de grande preocupação. A importância de manter um ambiente salubre é mais acentuada em estabelecimentos escolares (EE), tendo em conta, que no interior destes permanecem crianças durante um elevado período de tempo. É fundamental garantir uma boa QAI nos edifícios escolares, de forma a salvaguardar a saúde, o bem-estar e o conforto dos ocupantes, bem como, não comprometer o seu desempenho escolar. Recentemente, foram construídos novos edifícios escolares e alguns dos existentes foram alvo de obras de remodelação. Contudo, a crescente tendência em construir edifícios cada vez mais herméticos, com vista à diminuição dos gastos de energia, origina problemas como a reduzida ventilação dos espaços. Vários estudos têm demonstrado a influência das atividades de limpeza na QAI. No entanto, verifica-se que na maioria das escolas não existem ainda procedimentos de limpeza padronizados. A falta de instruções de trabalho e a ausência de formação às assistentes operacionais pode comprometer a eficácia dos procedimentos de higienização, o que poderá ter influência na QAI dos espaços. Este estudo teve como principal objetivo avaliar a QAI em escolas básicas de 1.º ciclo. Foram contemplados no estudo fatores como a tipologia do edifício, a ocupação das salas e as atividades de limpeza. Procedeu-se à caracterização dos EE e à monitorização de parâmetros ambientais, como a temperatura do ar, a humidade relativa, a velocidade do ar, o dióxido de carbono, o monóxido de carbono, as partículas, os microrganismos mesófilos totais e os fungos. Estes parâmetros foram avaliados nas salas com ocupação, sem ocupação e durante a implementação de um plano de higienização. A ventilação inadequada parece ser o fator que mais condiciona a QAI das salas de aula avaliadas. Registaram-se elevadas concentrações de dióxido de carbono e de microrganismos mesófilos totais, que parecem estar relacionados com a permanência dos ocupantes nos locais e com a falta de ventilação adequada dos espaços. A concentração de dióxido de carbono foi mais elevada em edifícios recentes. Os picos elevados na concentração de partículas parecem estar associados com as atividades dos ocupantes. Obtiveram-se concentrações menores de fungos e de microrganismos mesófilos totais ao longo da implementação do plano de higienização, o que poderá significar que os procedimentos de limpeza contribuem para reduzir os níveis de contaminação dos espaços interiores. No entanto, tendo em conta, que a concentração de microrganismos mesófilos totais permaneceu elevada, as operações de limpeza parecem não ser suficientes para garantir uma boa QAI. O aumento da ventilação dos espaços poderia contribuir significativamente para a melhoria da QAI dos espaços avaliados.

Avaliação da eficácia de produtos inovadores de proteção da madeira para exteriores

Nesta dissertação procurou-se comparar os desempenhos de três produtos de proteção da madeira quando expostos a várias condições de degradação. Tendo em consideração a grande quantidade de variáveis quando se faz este tipo de estudos, tentou-se controlar o maior número possível destas, de modo a permitir a comparação com outros estudos que venham a ser feitos futuramente. O objetivo a longo prazo é a contribuição desta dissertação para um conjunto de resultados cada vez mais abrangente, facilitando a escolha do utilizador, quando pretender proteger a madeira das agressões exteriores com este tipo de produtos. A principal conclusão nesta dissertação é que a agressividade do meio tem um papel determinante para os resultados obtidos, bem como as condições de inclinação e orientação da madeira. Situações de má selagem de topos ou não consideração da presença agentes biológicos são outros dois fatores que influenciam a durabilidade da construção. Finalmente, concluiu-se que não existem soluções únicas, tendo que se estudar caso a caso, quais as condições exatas de exposição, de forma a escolher o produto, ou produtos, mais adequados.

Beach sand and the potential for infectious disease transmission: observations and recommendations

Recent studies suggest that sand can serve as a vehicle for exposure of humans to pathogens at beach sites, resulting in increased health risks. Sampling for microorganisms in sand should therefore be considered for inclusion in regulatory programmes aimed at protecting recreational beach users from infectious disease. Here, we review the literature on pathogen levels in beach sand, and their potential for affecting human health. In an effort to provide specific recommendations for sand sampling programmes, we outline published guidelines for beach monitoring programmes, which are currently focused exclusively on measuring microbial levels in water. We also provide background on spatial distribution and temporal characteristics of microbes in sand, as these factors influence sampling programmes. First steps toward establishing a sand sampling programme include identifying appropriate beach sites and use of initial sanitary assessments to refine site selection. A tiered approach is recommended for monitoring. This approach would include the analysis of samples from many sites for faecal indicator organisms and other conventional analytes, while testing for specific pathogens and unconventional indicators is reserved for high-risk sites. Given the diversity of microbes found in sand, studies are urgently needed to identify the most significant aetiological agent of disease and to relate microbial measurements in sand to human health risk.