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Olive is recognized as a crop with great impact in agricultural, socioeconomic, environmental and public health sectors. The last is becoming more important during recent years as consequence of the increase of the pollen allergy in south Europe prompted by the widespread Olea pollen allergic reactions. The aim of the study was to quantify, for the first time, the variations of the Ole e 1 allergen amount in Olea pollen grains from four cultivars in three regions of Portugal. How weather parameters can affect the allergen production was also assessed. The study was conducted in three olive producer areas of Portugal from 2010 to 2013, Santarem (Central), Elvas (Southeast) and Mirandela (Tras-os-Montes region, Northeast). Mature pollen of four different cultivars (Cobrancosa, Arbequina, Picual and Verdeal) was collected during the olive flowering season. Ole e 1 was quantified using specific 2-site antibody ELISA. Pollen of the olive groves at the boundary Olea bioclimatic distribution in the Mirandela registered the higher allergen content for all varieties in each study year. Arbequina was the variety that showed the lower Ole e 1 allergen concentration, whereas the higher content was registered for Cobrancosa. The main meteorological parameters that influenced the allergen Ole e 1 concentration in the pollen grains were the rainfall and temperatures related variables. The knowledge of the allergenicity in different olive cultivars is an important tool in the selection of the most adequate for planting as ornamental crop and to adjust the pollen extracts used for diagnosis or even immunotherapy.

Air pollutants impact airborne pollen biochemistry. Oxidative damage to lipids, proteins and nucleic acids or protein nitration are among ozone (O3) and nitrogen dioxide (NO2) described deleterious effects possibly causing pollen physiology damage and enhanced allergenic activity, contributing to aggravate pollen driven respiratory allergy in urban areas. The goal of this research was to evaluate the effects of O3, NO2, alone and combined, on Dactylis glomerata pollen reactive oxygen species scavenging enzymes, on pollen germination and their potential contribution to the allergenicity. D. glomerata pollen was in vitro exposed to pollutants. Protein extracts were prepared and superoxide dismutase (SOD), catalase and glutathione peroxidase (GPx) activities were evaluated. Western blot with pooled sera or with IgG against group 5 allergens and profilin was performed. Pollen germination capacity was increased by NO2 and was unaffected by O3 or O3 + NO2 but showed longer pollen tubes in the latter. Exposure to O3 did not affect SOD activity but induced a twofold increase in catalase activity. SOD activity was twofold higher in pollen exposed to NO2. Exposure to O3 + NO2 induced a twofold and fivefold increase of SOD and catalase activities, respectively. Pollen GPx was unaffected by the pollutants. IgE-recognition of proteins in the molecular weight range of 42–57 kDa were amplified by NO2 and O3 + NO2 and O3 amplified proteins with molecular weight of 13 (profilin), 29 (Group 5), and 31 kDa. Taken together, these results show that pollen oxidative defences are activated by common air pollutants affecting both its germination capacity and its allergenic activity.

Pollen identification and quantification are used in many fields of application and research has been conducted to attain accurate automatic pollen recognition aiming to reduce the laborious work and subjectivity in human identification. The aim of our study was to evaluate the capacity of Raman parameters of pollen spectra, calculated for only 7 common band intervals in a limited spectral range, to be used as future technique in pollen automatic identification. There were analyzed 15 different pollen species considered to induce allergic reactions. Raman spectra were acquired at an excitation wavelength of 785 nm in a spectral region from 1000 to 1800 cm−1, preprocessed and deconvoluted to determine the Raman parameters: wavenumber, full width at half maximum of the band and integrated intensity. Seven common band intervals of all Raman spectra, in the fingerprint areas 1000–1010, 1300–1460 and 1500–1700 cm−1, were chosen for the classification of the pollen species using SVM (support vector machine). Our results showed that the classification accuracy of all pollen species was 100% in the training step, while in the testing step 14 out of the 15 pollen species were correctly assigned (93.3%), including the discrimination between 5 Poaceae species and between Betula pendula and Corylus avellana. It was also observed that all Raman parameters are important in the classification as well as all wavenumber areas considered. So, our study indicates that the Raman parameters of pollen spectra can be a promising methodology for automatic pollen recognition.

The target of this study was the tungsten Regoufe mine, whose exploitation stopped in the 1970s. When the mine closed, an unacceptable legacy constituted of mining waste tailings and the ruins of infrastructure was left behind. This work assessed the soil, plants, and water contamination in the mining area; namely their content in potentially toxic elements (PTEs). The global impact of PTEs in the Regoufe mine surface soil points to a very high to ultrahigh degree of contamination of the area having a serious ecological risk level, mainly related to As and Cd contributions. However, establishing the direct relation between As contamination and the anthropogenic effects caused by the mining process cannot be carried out in a straightforward manner, since the soils were already enriched in metals and metalloids as a result of the geological processes that gave origin to the mineral deposits. The studies performed on the plants revealed that the PTE levels in the plants were lower than in the soil, but site-specific soil concentrations in As and Pb positively influence bioaccumulation in plants. The magnetic studies showed the presence of magnetic technogenic particles concentrated in the magnetic fraction, in the form of magnetic spherules. The magnetic technogenic particles probably result from temperature increases induced by some technological process related to ore processing/mining activity. The PTEs in the surface and groundwater samples were similar and relatively low, being unlikely to pose potential health and environmental risks. Arsenic (As) constituted the exception, with levels above reference for drinking water purposes.

Particulate matter (PM) and pollen interaction, either airborne or at the respiratory mucosa needs further clarification, as allergic reaction intensification can be related to the PM physical characteristics and toxicity. This study aimed to investigate the physical-chemical properties of PM that can adhere to the pollen wall during its transport or inhalation, using Quercus spp. as a model, in three Portuguese cities with different geographical locations, meteorological influence and urbanization levels. Possible sources were evaluated through air masses trajectory analysis using the HYSPLIT model and correlation with meteorological factors. The sampling was performed using a 7-days Hirst-type volumetric sampler, and the pollen grains were observed using a Field Emission Electron Probe Microanalyser for PM analysis. A secondary electron image of each pollen grain was taken, to determine the adhered particles characteristics and energy dispersive x-ray spectroscopy (EDS) spectra were obtained for individual particles. A total of 484 pollen grains was observed, with 7683 particles counted and 1914 EDS spectra analyzed. The particle's equivalent diameter ranged from 0.3-16 μm, with most having a diameter3 μm. For the three cities, there were significant differences in the number of particles per pollen and the % area occupied by the particles. Particles adhered were mainly Si-rich, but variations in other dominant groups were observed. For Évora and Guarda, Ca-rich, SO-rich were second and third more representative, while Porto were Organic and Cl-rich. MetalsOxides were found in all cities with the highest number in Porto. P-rich particles were only found in Évora. Sea salt particles were observed in Évora, coincide with air mass trajectories possible carrying them from the Mediterranean Sea. In conclusion, the PM physical characteristics are similar between the studied cities, however, the dominant chemical composition is different, certainly impacting the exposome influence and pollen-allergy intensification towards the same pollen type and concentration.

In this work, we performed a palynological analysis of soil sampled from the footwear in order to provide their spatial and temporal classification. The sampling was performed in the shoes of 20 volunteers, in two different periods: autumn–winter and spring 2016/2017. The volunteers belonged to the same geographic region within a radius of 500 m. During the laboratorial treatment, the pollinic content present in the shoes sole was concentrated applying several treatments to destroy the exogenous material. A hierarchical cluster analysis was performed to verify whether it was possible to distinguish the samples based on their pollen assemblages. All the samples showed pollinic content, even when the shoes were apparently clean. The pollen types found were similar across the sampling period although with different pollen assemblages. In the winter samples, there was a prevalence of Fraxinus, Myrtaceae, Pinaceae (mostly Cedrus) and Poaceae pollen, while in the spring pollen of Olea, Poaceae, Pinaceae (namely Pinus) and Quercus was observed. Our results also demonstrated the discrimination between close sites (approximately 200 m), showing that the use of palynological analysis in footwear allows the collection of temporal and spatial information that can be used in forensic investigations.

Palynological, geochronological, and sedimentological analyses were conducted on samples from different depositional units of well-preserved marine, fluvial and palustrine sedimentary deposits from Vila Nova de Gaia located at the northwestern Portuguese coast. The studied deposits occur on two terraces: Terrace A—Lavadores (18 m to 33 m) and Terrace B—Canidelo (50 m). Chronological and sedimentological features allowed to date the deposits to the Upper Pleistocene until the last glacial period, being observed an evolution from a depositional setting in a marine intertidal zone, passing to a fluvial environment and ending in a palustrine/lagunar environment with possible periglacial influence. Pollen assemblages collected in the fluvial sedimentary horizons also suggested changes in the environmental conditions. A pollen content with good representation of arboreal species (Quercus, Pinus, Castanea, Fraxinus, Salix, Betula and Corylus), changed to a vegetation environment dominated by non-arboreal species, such as Poaceae, Plantago and Asteraceae. The top horizons, corresponding to a palustrine/lagunar environment, presented a pollen profile completely dominated by non-arboreal species, with minimal tree representation (Prunus, Quercus, Pinus and Castanea) which, combined with sedimentological characteristics may point to a more dry and cold climate than nowadays.

Platanus pollen is considered as a major cause of pollinosis in many southern European cities. This tree is commonly used in parks, avenues and gardens as ornamental species. The flowering period is short and abrupt between March and April, since this tree is characterized by having fast and explosive blooming process. This fact induces an important public health problem when Platanus bloom. The objective of the study is to evaluate the pollen and allergen concentration in two cities in the north-west of the Iberian Peninsula, Porto (Portugal) and Ourense (Spain), and analyse how weather parameters and environmental pollutants can affect the patterns of pollen and allergen airborne distribution. The pollen sampling was conducted, during 2018, using a Hirst volumetric sampler, and Pla a 1 aeroallergen was collected using a Burkard multivial cyclone sampler. The pollen counting was obtained following the Spanish Aerobiology Network methodology. Aeroallergens were quantified by ELISA method using a Pla a 1-specific antibody. Platanus pollen concentrations are seven times lower in the Porto atmosphere than in Ourense, but the Pla a 1 content is only 2 times higher in Ourense city. These differences can be explained by the influence of air pollutants in plants and meteorological factors. Platanus airborne pollen and Pla a 1 aeroallergen were correlated with some meteorological parameters, namely temperature, rainfall and relative humidity, and with atmospheric pollutants such as NO2 and O3. The results suggest the existence of different allergenic activities in the atmosphere of both cities.

The knowledge of pollen sensitivity and tolerance to stress factors such as air pollution is important for forest sustainability, ensuring the most efficient production with the highest benefits and lowest resource losses. This study intended to evaluate the influence of common air pollutants in four forest trees species, Betula pendula Roth, Corylus avellana L., Acer negundo L. and Quercus robur L., through a comparative analysis at the same experimental conditions. We aimed to investigate the effect that may occur in pollen fertility, protein content, oxidative stress and wall composition after exposure in vitro to ozone and nitrogen dioxide at concentration levels for vegetation protection in Europe. Our results suggest changes in pollen viability, protein content and differential sensitivity related to ROS synthesis, NADPH oxidase activity, as well as in wall composition. The results indicate that NO2 exposure affected more the pollen species studied mostly at the highest concentration exposure. As for ozone, there were less significant differences between samples, however, a different behavior occurs in O3 expositions, where the most influence happens at the legal limit for vegetation protection in Europe. Our study showed that significant pollen functions could be compromised even at common air pollutant&rsquo, s concentrations.

The aim of this study was to assess the thermal requirements of the most important grapevine varieties in northwestern Spain to better understand the impact of climate change on their phenology. Different phenological models (GDD, GDD Triangular and UniFORC) were tested and validated to predict budburst and flowering dates of grapevines at the variety level using phenological observations collected from Treixadura, Godello, Loureira and Albariño between 2008 and 2019. The same modeling framework was assessed to obtain the most suitable model for this region. The parametrization of the models was carried out with the Phenological Modeling Platform (PMP) platform by means of an iterative optimization process. Phenological data for all four varieties were used to determine the best-fitted parameters for each variety and model type that best predicted budburst and flowering dates. A model calibration phase was conducted using each variety dataset independently, where the intermediate-fitted parameters for each model formulation were freely-adjusted. Afterwards, the parameter set combination of the model providing the highest performance for each variety was externally validated with the dataset of the other three varieties, which allowed us to establish one overall unique model for budburst and flowering for all varieties. Finally, the performance of this model was compared with the attained one while considering all varieties in one dataset (12 years × 4 varieties giving a total number of observations of 48). For both phenological stages, the results showed no considerable differences between the GDD and Triangular GDD models. The best parameters selected were those provided by the Treixadura GDD model for budburst (day of the year (t0) = 49 and base temperature (Tb) = 5) and those corresponding to the Godello model (t0 = 52 and Tb = 6) for flowering. The modeling approach employed allowed obtaining a global prediction model that can adequately predict budburst and flowering dates for all varieties. Ministerio de Agricultura, Pesca y Alimentación (España) | Ref. 20190020007473 Xunta de Galicia | Ref. ED431C 2017/62-GRC Xunta de Galicia | Ref. ED481A-2017/209 Fundação para a Ciência e a Tecnologia (Portugal) | Ref. FCT UIDB / 04683/2020 Fundação para a Ciência e a Tecnologia (Portugal) | Ref. UIDP / 04683/2020

Natural forests are considered a reservoir of great biological diversity constituting one of the most important ecosystems in Europe. Quercus study is essential to assess ecological conservation of forests, and also of economic importance for different industries. In addition, oak pollen can cause high sensitization rates of respiratory allergies in pollen-allergy sufferers. This study sought to know the pollen production of six oak species in the transitional area between the Eurosiberian and Mediterranean Bioclimatic Regions, and to assess the impact of climate change on airborne oak pollen concentrations. The study was conducted in Ourense (NW Spain) over the 1993&ndash, 2019 period. A Lanzoni VPPS 2000 volumetric trap monitored airborne pollen. A pollen production study was carried out in ten trees randomly selected in several Quercus forest around the Ourense city. Oak pollen represented around 14% of annual total pollen registered in the atmosphere of Ourense, showing an increasing trend during the last decade. Pollen production of the six studied oak species follow the proportions 1:1:2:5:90:276 for Q. ilex, Q. faginea, Q. rubra, Q. suber, Q. pyrenaica, and Q. robur respectively. We detected a significant trend to the increase of the annual maximum temperature, whereas a decrease of the maximum and mean temperatures during three previous months to oak flowering. This could be related with the detected trend to a delay of the oak Main Pollen Season onset of 0.47 days per year. We also found significant trends to an increase of the annual pollen integral of 7.9% pollen grains per year, and the pollen peak concentration of 7.5% pollen grains per year. Quercus airborne pollen monitoring as well as the knowledge of the reproductive behavior of the main oak species, bring us an important support tool offering a promising bio-indicator to detect ecological variations induced by climate change.

Phenological, aerobiological, and weather data are useful tools to study local and regional flowering dynamics in crops with economic importance. The present study focuses on four autochthonous grapevine cultivars, namely, &lsquo, Treixadura&rsquo, &lsquo, Godello&rsquo, Loureira&rsquo, and &lsquo, Albari&ntilde, o&rsquo, (Vitis vinifera L.), which belong to the Designation of Origin Ribeiro area (located in northwestern Spain) from 2015&ndash, 2019. The aims of the work were to (1) compare the airborne pollen concentration in the vineyard collected by two different traps, (2) analyze the influence of the main meteorological variables on cultivar phenology and pollen concentration, and (3) test the contribution of the air masses on pollen concentrations in the vineyard. Phenological development has been assessed twice weekly, according to the Biologische Bundesanstalt, Bundessortenamt und Chemische Industrie (BBCH) scale. Airborne pollen concentrations were monitored by using two traps during stage 6 (flowering), namely, a Hirst volumetric sampler and a Cour passive trap. The bioclimatic conditions affected the duration of flowering, ranging from 11 and 24 days. The highest seasonal pollen integral (SPIn) was registered in 2016 for the Hirst sampler, with 302 pollen, and in 2019 for the Cour trap, with 1,797,765 pollen/m2/day. The main variables affecting pollen concentrations were average temperature during the main pollen season, as well as, temperatures and dew points during the pre-peak period. The relationship between pollen data registered by both traps and the obtained harvest indicate that the Hirst trap may be more suitable for predicting a local production and that the Cour sampler is more appropriate for forecasting regional productions.

In the last years, pollen-related respiratory allergies have increased worldwide. In the case of Cupressaceae pollen, allergy symptoms appearing during the winter are often confused with generic respiratory diseases such as the common cold, influenza syndrome, among others. The aim of our study was to monitor the atmospheric of Cupressaceae pollen in the city of Porto, investigate its diurnal variation, as well as find the meteorological factors influencing its atmospheric concentration. Airborne pollen sampling was performed using a 7-day Hirst-type volumetric spore trap for 5 years (2013–2017). Cupressaceae main pollen season, interannual and diurnal variations, as well as correlations with the meteorological parameters were determined. During the study period, 2015 was the year that registered the highest annual pollen concentration, while 2016 presented the lowest. Cupressaceae pollen season occurred during the winter, being February the month with the highest concentration. The diurnal distribution pattern presents a well-defined peak concentration in the morning hours (9 a.m.–12 a.m.), which decays abruptly in the early afternoon. Finally, airborne pollen concentration and meteorological factors presented significant correlations, with temperature being the most influent variable.

In the context of the forensic sciences, geochemical data can give important information as to the sample origin or help to refine and narrow the probable origin. The current study employs ...

Phenological models for predicting the grapevine flowering were tested using phenological data of 15 grape varieties collected between 1990 and 2014 in Vinhos Verdes and Lisbon Portuguese wine regions. Three models were tested: Spring Warming (Growing Degree Days – GDD model), Spring Warming modified using a triangular function – GDD triangular and UniFORC model, which considers an exponential response curve to temperature. Model estimation was performed using data on two grape varieties (Loureiro and Fernão Pires), present in both regions. Three dates were tested for the beginning of heat unit accumulation (t0 date): budburst, 1 January and 1 September. The best overall date was budburst. Furthermore, for each model parameter, an intermediate range of values common for the studied regions was estimated and further optimized to obtain one model that could be used for a diverse range of grape varieties in both wine regions. External validation was performed using an independent data set from 13 grape varieties (seven red and six white), different from the two used in the estimation step. The results showed a high coefficient of determination (R2: 0.59–0.89), low Root Mean Square Error (RMSE: 3–7 days) and Mean Absolute Deviation (MAD: 2–6 days) between predicted and observed values. The UniFORC model overall performed slightly better than the two GDD models, presenting higher R2 (0.75) and lower RMSE (4.55) and MAD (3.60). The developed phenological models presented good accuracy when applied to several varieties in different regions and can be used as a predictor tool of flowering date in Portugal.

Pollen quality of 15 cultivars of Vitis vinifera L. was studied in this work. Pollen viability was tested by the fluorochromatic reaction and germination was analyzed by in vitro assays, using two different media. Differences among cultivars in the number of pollen apertures were observed under light microscope. All the cultivars studied showed a higher percentage of tricolporated pollen, however, pollen grains containing one, two or four apertures were also observed. The cultivar Loureiro was the one with the higher percentage of pollen grains with four apertures (3.8%) and Touriga Nacional presented 100% of tricolporated pollen grains. The viability analysis showed that 13 cultivars presented values higher than 50%, with 8 cultivars reaching values above 75%. The pollen germination rates vary greatly for the grapevine cultivars studied, three cultivars show low values of germination (under 14%) in the two media tested, which were Touriga Nacional, Cabernet Franc, and Cabernet Sauvignon while others presented high values of germination like Castelao, Loureiro, Malbec and Petit Verdot. No significant statistical differences between the percentages of germination in the two media studied were found for the majority of cultivars analyzed.

In the present study, the effects of two important oxidizing atmospheric pollutants (O3 and NO2) on the allergenic properties and chemical composition of Platanus x acerifolia pollen were studied. Pollen samples were subjected to O3 and/or NO2 under in vitro conditions for 6h at atmospheric concentration levels (O3: 0.061ppm; NO2: 0.025ppm and the mixture of O3 and NO2: 0.060 and 0.031ppm respectively). Immunoblotting (using Pla a 1 and Pla a 2 antibodies), infrared and X-ray photoelectron spectroscopy techniques were used. Immunochemical analysis showed that pollen allergenicity changes were different according to the pollutant tested (gas or mixture of gasses) and that the same pollutant gas may interact in a different manner with each specific allergen. The spectroscopy results showed modifications in the FTIR spectral features of bands assigned to proteins, lipids, and polysaccharides of the pollen exposed to the pollutants, as well as in the XPS spectra high-resolution components C 1s, N 1s, and O 1s. This indicates that while airborne, the pollen wall suffers further modifications of its components induced by air pollution, which can compromise the pollen function.

Olive trees are one of the most economically important perennial crops in Portugal. During the last decade, the Alentejo olive-growing region has suffered a significantly change in the crop production system, with the regional pollen index (RPI) and olive fruit production registering a significant growth. The aim of this study was to ascertain the utility of this highly variable production and pollen data in crop forecasting modeling. Airborne pollen was sampled using a Cour-type trap from 1999 to 2015. A linear regression model fitted with the regional pollen index as the independent variable showed an accuracy of 87% in estimating olives fruit production in Alentejo. However, the average deviation between observed and modeled production was 32% with half of the tested years presenting deviations between 36 and 66%. The low accuracy of this model is a consequence of the great overall variation and significant upward trend observed in both the production and the RPI dataset that conceal the true association between these variables. In order to overcome this problem, a detrend procedure was applied to both time series to remove the trend observed. The regression model fitted with the fruit production and the RPI detrended data showed a lowest forecasting accuracy of 63% but the average deviation between observed and modeled production decrease to 14% with a maximum deviation value of 33%. This procedure allows focusing the analysis on the production fluctuations related to the biological response of the trees rather than with the changes in the production system.

A wine forecast model for one of the most arid wine regions of the Europe—Alentejo was improved and tested for the period 1998–2014. During this period, Alentejo region had strong upward trends in wine production associated to the increase of vineyard area. The forecast model was supported on a hierarchical analysis, including the determination of the potential production at flowering by quantifying airborne pollen concentration, followed by a climate based evaluation of the possible impact of fruit-set conditions in the limitation of production. Through the monitoring of airborne pollen flows it is possible to define an accurate main pollen season and determine the regional pollen index that will be used as independent variable in the regional forecast model. The time trend, which was initially removed from data, was then added back to obtain the forecast. Stepwise regression and cross-validation were employed during the period 1998–2014 for calibration of the model used for predicting annual wine production. The developed model explained about 86% of wine variance over the years with absolute average error of 6% for the cross validation and 87% of cases had differences between actual and forecasted wine production below 10%. The reliability and early-indication ability of the proposed forecast model justify their use to respond to a number of government agencies and wine industry concerns and activities.

The knowledge of the allergen content in the atmosphere is a useful tool to stablish the risk allergy warnings for the sensitive people. In Portugal the main airborne allergenic pollen come from trees (such as Betula or Olea), grasses or weeds (mainly Urticaceae). The present study sought the quantification of the Bet v 1, Ole e 1, Lol p1 and Par j1–2 aeroallergen concentration as well as how weather variables influence in the pollen and allergen concentration in Porto city. Aerobiological study was carried out by a Hirst-type volumetric sampler for pollen collection and a Burkard Cyclone sampler for the aeroallergens. A regression analysis between pollen and allergens was conducted for the identification the allergenic risk days. High Pollen Allergen Potency in the atmosphere was observed considering the low levels of airborne pollen detected. A significant and positive correlation has been obtained between pollen and aeroallergen values with the temperatures whereas the correlation was negative with relative humidity, rainfall and wind speed. Back trajectory methodology was applied in order to analyse the discordances between pollen and allergen maximum concentrations. The analysis showed that when the pollen and allergen peaks were registered on the same day, air masses always comes from the continent. However, when the peaks do not coincide, the air mass comes from the continent in the case of the pollen peak and from the sea for the allergen peak. This behaviour can be a consequence of the high humidity in the air masses from the sea, which can benefit the allergen release from pollen grains. In our study it was observed that the available traditional information for allergenic Type I patients, corresponding to the amount of pollen grains in the bioaerosol, do not accurately identify the real allergenic load in the air.

Pollen can be modified by the increasing environmental pollution which can lead to a rising trend in pollen-induced respiratory allergies. The aim of this work was to study the fertility, protein profile and allergenicity of Plantago lanceolata and Salix atrocinerea pollen after exposure to ozone. The pollen collected directly from the anthers of both species was exposed to O3 in an environmental chamber, for 6 h at different concentrations. After exposing the pollen, viability tests and in vitro germination were performed. Total protein content was determined by the Bradford method, the polypeptides were separated by SDS-PAGE and the antigenic properties were analyzed by immunological techniques using patients’ sera. The concentrations of O3 tested affected the pollen fertility and the total protein content, but the protein profiles did not show any differences. Furthermore, the immunological tests revealed an increase in IgE binding reactivity when pollen was exposed to the pollutant.

In the present study, the effects of two important oxidizing atmospheric pollutants (O

Atmospheric gaseous pollutants can induce qualitative and quantitative changes in airborne pollen characteristics. In this work, it was investigated the effects of carbon dioxide (CO2) on Acer negundo pollen fertility, protein content, allergenic properties, and carbohydrates. Pollen was collected directly from the anthers and in vitro exposed to three CO2 levels (500, 1000, and 3000 ppm) for 6 and 24 h in an environmental chamber. Pollen fertility was determined using viability and germination assays, total soluble protein was determined with Coomassie Protein Assay Reagent, and the antigenic and allergenic properties were investigated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) and immunological techniques using patients' sera. Also, pollen fructose, sucrose, and glucose values were determined. Carbon dioxide exposure affected negatively pollen fertility, total soluble protein content, and fructose content. The patient sera revealed increased IgE reactivity to proteins of A. negundo pollen exposed to increasing levels of the pollutant. No changes were detected in the SDS-PAGE protein profiles and in sucrose and glucose levels. Our results indicate that increase in atmospheric CO2 concentrations can have a negative influence of some features of A. negundo airborne pollen that can influence the reproductive processes as well as respiratory pollen allergies in the future.

Airborne pollen emission model was used to determine pollen metrics and to examine their relationship with vineyard phenology in two wine regions of Northern Portugal: Vinhos Verdes (1993-2007) and Douro (1992-2011). A number of airborne pollen metrics were obtained through the rate of changes of logistic model adjusted to the time series of airborne pollen. In both regions, the mean absolute differences between observed phenology and model- predicted values for start, peak and final of flowering phenophases were always lower than 5 days and the slope of the regression through the origin is close to one. These metrics can be used to accurately and precisely predict the dynamic of Vitis flowering observed at field level. The model's simplicity and flexibility are of great advantage for its practical use in aerobiology.

Airborne pollen calendars are useful to estimate the flowering season of the different plants as well as to indicate the allergenic potential present in the atmosphere at a given time. In this study, it is presented a 10-year survey of the atmospheric concentration of allergenic pollen types. Airborne pollen was performed, from 2003 to 2012, using a 7-day Hirst-type volumetric trap. The interannual variation of the daily mean concentration of the number of pollen grains and the main pollen season was determined as well as the hourly variations and correlation with meteorological parameters. During the study period, 18 different allergenic pollen types were considered based on its representativeness on the total annual airborne pollen concentration. The lowest annual concentrations were sampled in 2006 and the highest in 2007. The highest airborne pollen concentration was found during early spring and early summer. On the contrary, December was the month with the lowest pollen concentration. The major pollen sampled belongs to trees followed by weeds and grasses, being the most representative pollen types in the atmosphere: Urticaceae, Platanus, Poaceae, Pinaceae, Cupressaceae, Acer, Quercus, Castanea, Plantago, Alnus, Olea europaea, Betula, Myrtaceae and Populus. Intradiurnal distribution patterns of the pollen types studied presented differences with some taxa being predominantly sampled in the morning (9–11 a.m.) while others in first night hours (between 9 and 12 p.m.). Significantly correlations were found between the airborne pollen concentration and meteorological parameters.

Pollen of Betula pendula, Ostrya carpinifolia and Carpinus betulus was exposed in vitro to two levels of NO2 (about 0.034 and 0.067 ppm) – both below current atmospheric hour-limit value acceptable for human health protection in Europe (0.11 ppm for NO2). Experiments were performed under artificial solar light with temperature and relative humidity continuously monitored. The viability, germination and total soluble proteins of all the pollen samples exposed to NO2 decreased significantly when compared with the non-exposed. The polypeptide profiles of all the pollen samples showed bands between 15 and 70 kDa and the exposure to NO2 did not produce any detectable changes in these profiles. However, the immunodetection assays indicated higher IgE recognition by patient sera sensitized to the pollen extracts from all exposed samples in comparison to the non-exposed samples. The common reactive bands to the three pollen samples correspond to 58 and 17 kDa proteins.

Raman microspectroscopy allows a non-destructive identification of airborne particles. However, the identification of particles such as pollen is hindered by the absence of a spectral library. Although reference spectra of pollen have been published before, they have always been limited to a certain number of species. In this work, Raman spectra of 34 pollen types are presented and were used to build a pollen spectra primary library. Afterward, the applicability of this database for detecting and identifying pollen in airborne samples was tested. Airborne pollen samples collected during April, May and August were compared with blank pollen spectra by means of Hit Quality Index. Although a much larger library would be required, our results showed that all first hits correspond to the same blank pollen species of the questioned sample from the air. This possibility is an innovative idea and a promising line of investigation for future RAMAN technology development in the area of aerobiology.

This study aims to investigate the effects of O3 in protein content and immunoglobulin E (IgE)-binding profiles of Acer negundo, Platanus x acerifolia and Quercus robur pollen. Pollen was exposed to O3 in an environmental chamber, at half, equal and four times the limit value for the human health protection in Europe. Pollen total soluble protein was determined with Coomassie Protein Assay Reagent, and the antigenic and allergenic properties were investigated by SDS-PAGE and immunological techniques using patients' sera. O3 exposure affected total soluble protein content and some protein species within the SDS-PAGE protein profiles. Most of the sera revealed increased IgE reactivity to proteins of A. negundo and Q. robur pollen exposed to the pollutant compared with the non-exposed one, while the opposite was observed in P. x acerifolia pollen. So, the modifications seem to be species dependent, but do not necessarily imply that increase allergenicity would occur in atopic individuals.

Platanus hispanica pollen is considered an important source of aeroallergens in many Southern European cities. This tree is frequently used in urban green spaces as ornamental specie. The flowering period is greatly influenced by the meteorological conditions, which directly affect its allergenic load in the atmosphere. The purpose of this study is to develop equations to predict the Platanus allergy risk periods as a function of the airborne pollen, the allergen concentration and the main meteorological parameters. The study was conducted by means two volumetric pollen samplers; a Lanzoni VPPS 2000 for the Platanus pollen sampling and a Burkard multivial Cyclone Sampler to collect the aeroallergen particles (Pla a_1). In addiction the Dot-Blot and the Raman spectroscopy methods were used to corroborate the results. The Pla a_1 protein is recorded in the atmosphere after the presence of the Platanus pollen, which extend the Platanus pollen allergy risk periods. The Platanus pollen and the Pla a 1 allergens concentration are associated with statistical significant variations of some meteorological variables: in a positive way with the mean and maximum temperature whereas the sign of the correlation coefficient is negative with the relative humidity. The lineal regression equation elaborated in order to forecast the Platanus pollen content in the air explain the 64.5% of variance of the pollen presence in the environment, whereas the lineal regression equation elaborated in order to forecast the aeroallergen a 54.1% of the Pla a_1 presence variance. The combination of pollen count and the allergen quantification must be assessed in the epidemiologic study of allergic respiratory diseases to prevent the allergy risk periods.

Betula pendula pollen, under laboratory conditions, was exposed to three atmospheric pollutants: carbon monoxide (CO), ozone (O3) and sulphur dioxide (SO2). Two levels of each pollutant were used; the first level corresponds to a concentration on the atmospheric hour-limit value acceptable for human health protection in Europe, the second level to a higher, at least more than double of the first, concentration level. Experiments were done under artificial solar light with controlled temperature and relative humidity. Our results indicate that, in urban areas, concentrations of CO, O3 and SO2 on the limits established for human protection, can affect pollen fertility. We verified a decrease in the viability and germination of the pollen, indicating damage to the pollen membrane system. Also, a general decreasing trend in the total protein content of the exposed samples when compared with the control samples was observed, which suggests alterations in the antigenic characteristics of pollen.

Ostrya spp. and Carpinus spp. pollen was in vitro exposed to three atmospheric pollutants: CO, O3 and SO2. Two levels of each pollutant were used, and the first level corresponds to a concentration about the atmospheric hour-limit value acceptable for human health protection in Europe and the second level to about the triple of the first level. Experiments were done under artificial solar light with temperature and relative humidity controlled. The viability of the exposed pollen samples showed a significant decrease. Also, the germination percentage showed a significant decrease in both exposed pollens, and the effect was most pronounced for SO2, followed by O3 and CO. A general decreasing trend in the total soluble protein content of the exposed pollen samples when compared with the control was observed, but it was only statistically significant for the Ostrya spp pollen. The results showed marked effects were observed on the Ostrya spp. and Carpinus spp. pollen when exposed to air pollutant levels that can be considered safe for human health protection.

In this study, the effect of three different ozone (O3) levels on soluble protein content of Acer negundo, Quercus robur and Platanus spp. pollen was investigated. Pollen was directly collected from each plant and exposed to O3 values below, equal and four times the limit for human health protection in an environmental chamber system. With this chamber, it was possible to control and reproduce the O3 levels between different experiments. Total protein content was determined colorimetrically with Coomassie protein assay reagent (Pierce) by the Bradford method. Results showed that O3 has significant effects on pollen causing a decrease in soluble proteins that seem to be dependent on species and gas concentration. Also, the existence of reproducibility in the effects of O3, within the same concentration, on the protein content of pollen was verified.

Cladosporium spores are ubiquitous in the air and are included in the fungal contaminants that define environmental quality. This paper provides an updated review of their concentration in the Iberian Peninsula, based on data from 12 sampling stations collected using a common non-viable volumetric sampling method and the same subsequent data-treatment techniques. The database comprised 84 annual periods, covering the period 1993–2009. Mean annual total spore concentrations for each station, calculated as the sum of daily concentrations, ranged from 54 459 in Cartagena to 933 485 in Sevilla; other sampling stations recorded annual total spores between 93 052 (Porto) and 579 953 (Merida). Analysis of annual spore-distribution patterns revealed either one or two peaks (spring and fall) depending on the location and prevailing climate of the area where the sampling stations have been placed. For all stations, average temperature was the meteorological parameter displaying the strongest positive corre...

The Algarve is located at a very short distance from North Africa, in Southern Portugal, and as one of the most touristic regions of Portugal, it is accessible by air, land and sea. It is very susceptible to many illegal activities, such as illegal migration, drug trafficking, kidnapping, and murder, among others. Therefore, an Algarve soils database for forensic purposes is being conducted with the conjunction of geological and palynological methodologies on soils characterization, since this is of fundamental importance to assess reliable evidence on forensic investigations. In this study, the properties of soils from several proximate sites from the Algarve were investigated, namely: (i) colour determined by spectrophotometry; (ii) particle size distribution determined by laser granulometry; (iii) low-field magnetic susceptibility by a susceptibility meter; and (iv) pollen content using a light microscope. Finally, a hierarchical cluster analysis was applied to ascertain the capacity of the different soil properties for discrimination between samples. The study reveals the utility of geobotanical techniques for forensic discrimination of soils. Even though some similarities between some of the samples were found, each one presented a combination of colour, particle size distribution, magnetic susceptibility and pollen features that enable the determination of a fingerprint expected to reveal a specific site for future selection of coastal search areas in the Algarve region.

The aim of this work was to identify the main allergy-related Ascomycetes fungal spores present in the atmosphere of Porto, using different and complementary techniques. The atmospheric sampling, performed in the atmosphere of Porto (Portugal) from August 2006 to July 2008, indicated Cladosporium, Penicillium, Aspergillus and Alternaria as the main fungal spore taxa. Alternaria and Cladosporium peaks were registered during summer. Aspergillus and Penicillium highest values were registered from late winter to early spring. Additionally, the Andersen sampler allowed the culture and isolation of the collected viable spores subsequently used for different identification approaches. The internal-transcribed spacer region of the nuclear ribosomal repeat unit sequences of airborne Ascomycetes fungi isolates revealed 11 taxonomically related fungal species. Among the identified taxa, Penicillum and Aspergillus presented the highest diversity, while only one species of Cladosporium and Alternaria, respectively, were identified. All selected fungal spore taxa possessed phosphatase, esterase, leucine arylamidase and beta-glucosidase enzymatic activity, while none had lipase, cystine arylamidase, trypsin or beta-glucuronidase activity. The association between the spore cell wall morphology, DNA-based techniques and enzymatic activity approaches allowed a more reliable identification procedure of the airborne Ascomycota fungal spores.

SUMMARYThe aim of the present study was to develop a hierarchical bioclimatic model for forecasting olive crop yields in the Alentejo region of south-eastern Portugal. The model was estimated for three different developmental stages: (1) at flowering, using only the regional pollen index (RPI); (2) at fruit growth using RPI and a plant water requirements index (PWRI) and (3) at fruit maturing using RPI plus a water requirements index plus a phytopathological index (PPI). Olive airborne pollen was sampled from 1999 to 2007, using a Cour trap installed in Reguengos de Monsaraz. The meteorological parameters used in the calculation of the post-flowering indices corresponded to data from a meteorological station located near the airborne sampling point. At the flowering stage, 0·66 of the regional olive yield can be explained by the RPI with an average deviation between observed and predicted production of 0·15 for the forecast model internal validation and of 0·19 for the cross-validation. The addition of the variable PWRI to the forecasting model explained an additional 0·26 of the variation, while the PPI explained an additional 0·05. The final bioclimatic model, with all the three variables tested, explained 0·97 of the regional olive fruit yield being the average deviation between observed and predicted production of 0·04 for the internal validation of the model and of 0·07 for the external validation. The hierarchical nature of this bioclimatic model, along three different development stages, enabled the prediction to be updated as the growing season progressed.

Trees are considered producers of allergenic pollen. The aims of this work were to characterize the aerobiology of the Platanus, Acer, Salix, Quercus, Betula and Populus pollen, linking it with monthly emergency hospital admissions and to identify the different reactivity levels in sensitized patients. This information would be of great importance to evaluate the convenience of changing the inventory of pollen producer trees related to the risk of allergenic reactions. The study was conducted in Porto, Portugal, from 2005 to 2007. Airborne pollen was sampled using a Hirst-type volumetric trap. The antigenic and allergenic properties of Acer negundo, Betula pendula, Platanus occidentalis, Populus hybrida, Quercus robur and Salix babylonica pollen, collected in public gardens or sidewalks, were investigated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis (SDS-PAGE) and immunological techniques using polysensitized-patient sera. Monthly hospital admissions of asthma or dyspnea related with respiratory diseases were obtained from the Emergency Room database of Hospital Geral de Santo António. Tree pollen and hospital admissions were positively correlated. Tree pollen peaked in March which coincides with the hospital admissions maximum. The highest binding affinity was observed with A. negundo, S. babylonica and P. occidentalis pollen extracts and the lowest with P. hybrida. Consistently, Acer and Platanus maximum airborne pollen concentrations were observed during March attaining levels considered moderate to high risk for allergenic reactions. Prominent bands with approximately 71, 35, 31, 22, 19, 16, 14, 13 and 11 kDa were revealed. A 52 kDa band was shared by all analyzed sera. High levels of airborne pollen and emergency hospital admissions were related. High binding affinity of specific IgE to pollen extracts of the most abundant tree pollen present in the atmosphere was observed. Patient sera revealed multiple similar allergenic bands shared by the different extracts. This multidisciplinary approach is useful in day-to-day medical practice to help in diagnostic, therapeutic and allergy alerting system adjusting.

Because fungal spore emission intensity varies throughout the year and even throughout the day, study of the hourly distribution is an important aspect of aerobiological monitoring. The objective of this work was to determine seasonal and intradiurnal variation of allergenic airborne fungal spores in urban and rural areas. The aerobiological study was performed from 2005 to 2007 using Hirst-type volumetric spore traps. Fungal spore types (Cladosporium, Alternaria, and Aspergillus/Penicillium) were selected according to their allergenic capacity and their frequency in the atmosphere. Cladosporium was the most frequent fungal spore in both locations, together with Alternaria; its concentration was higher during summer whereas Aspergillus/Penicillium was more abundant during autumn. Alternaria and Cladosporium maximum daily spore concentration was reached from 16.00 to 20.00 h. Aspergillus/Penicillium spore distribution differed in the two locations, and although it was difficult to establish a distribution pattern the highest concentration were found during the night. Determination of periods of high concentration may help allergic patients to avoid the daily periods when the allergen concentration is highest.

In this work, the structure and protein profile of Chenopodium alba pollen samples collected in rural and urban environments were compared. Pollen structure was studied using light and scanning electron microscopy and the soluble protein profile was investigated by means of biochemical analyses (SDS-PAGE). The rural pollen has the opercules well defined while in the urban pollen there is a fine film covering its wall, and opercules are deformed. A decrease in the protein content in the urban pollen and different protein profiles were observed. Micro-Raman spectroscopic studies allowed the identification of carbon-containing particles in the urban pollen wall as well as its morphology and size. The results of this study indicate that the particles present in the urban air are diesel exhaust particles that are adsorbed and accumulated on the Chenopodium alba pollen. These particles can contribute for modifying their structure and protein content.

Although fungal spores are an ever-present component of the atmosphere throughout the year, their concentration oscillates widely. This work aims to establish correlations between fungal spore concentrations in Porto and Amares and meteorological data. The seasonal distribution of fungal spores was studied continuously (2005-2007) using volumetric spore traps. To determine the effect of meteorological factors (temperature, relative humidity and rainfall) on spore concentration, the Spearman rank correlation test was used. In both locations, the most abundant fungal spores were Cladosporium, Agaricus, Agrocybe, Alternaria and Aspergillus/Penicillium, the highest concentrations being found during summer and autumn. In the present study, with the exception of Coprinus and Pleospora, spore concentrations were higher in the rural area than in the urban location. Among the selected spore types, spring-autumn spores (Coprinus, Didymella, Leptosphaeria and Pleospora) exhibited negative correlations with temperature and positive correlations both with relative humidity and rainfall level. On the contrary, late spring-early summer (Smuts) and summer spores (Alternaria, Cladosporium, Epicoccum, Ganoderma, Stemphylium and Ustilago) exhibited positive correlations with temperature and negative correlations both with relative humidity and rainfall level. Rust, a frequent spore type during summer, had a positive correlation with temperature. Aspergillus/Penicillium, showed no correlation with the meteorological factors analysed. This knowledge can be useful for agriculture, allowing more efficient and reliable application of pesticides, and for human health, by improving the diagnosis and treatment of respiratory allergic disease.

In this work the objective was to develop a bioclimatic model to forecast olive yield based on airborne pollen, soil water content, and favourable conditions for phytopathological attacks. Olive airborne pollen was sampled from 1998 to 2006 using Cour traps installed in the Tras-os-Montes e Alto Douro region, in the provinces of Valenca do Douro and Vila Nova de Foz-Coa. Meteorological data from a meteorological station located in Pinhao, near the pollen samplers, was used to calculate other independent variables. According to the bioclimatic model, at the flowering stage 63% of regional olive production can be predicted from the regional pollen index, with an average deviation between observed and predicted production of 10%. The variable soil water content enabled an increase in forecasting accuracy of about 30%, and a reduction in the average deviation between observed and predicted production of 6%. The final regression with all three variables tested showed that the bioclimatic model was able to predict the annual variability of regional olive fruit production with an accuracy of 97%, the average deviation between observed and predicted production being 3% for internal validation and 6% for external validation.