Dew point temperature affects ascospore release of allergenic genus Leptosphaeria.

The genus Leptosphaeria contains numerous fungi that cause the symptoms of asthma and also parasitize wild and crop plants. In search of a robust and universal forecast model, the ascospore concentration in air was measured and weather data recorded from 1 March to 31 October between 2006 and 2012. The experiment was conducted in three European countries of the temperate climate, i.e., Ukraine, Poland, and the UK. Out of over 150 forecast models produced using artificial neural networks (ANNs) and multivariate regression trees (MRTs), we selected the best model for each site, as well as for joint two-site combinations. The performance of all computed models was tested against records from 1 year which had not been used for model construction. The statistical analysis of the fungal spore data was supported by a comprehensive study of both climate and land cover within a 30-km radius from the air sampler location. High-performance forecasting models were obtained for individual sites, showing that the local micro-climate plays a decisive role in biology of the fungi. Based on the previous epidemiological studies, we hypothesized that dew point temperature (DPT) would be a critical factor in the models. The impact of DPT was confirmed only by one of the final best neural models, but the MRT analyses, similarly to the Spearman's rank test, indicated the importance of DPT in all but one of the studied cases and in half of them ranked it as a fundamental factor. This work applies artificial neural modeling to predict the Leptosphaeria airborne spore concentration in urban areas for the first time.

A comparative study of hourly and daily relationships between selected meteorological parameters and airborne fungal spore composition.

Air sampling was conducted in Szczecin (Poland) throughout April-September 2013. The final data set included 177 daily and 4248 hourly samples. The total of 21 types of spores, which occurred in a number >10 in the season, were taken into account. The following meteorological parameters were analyzed: air temperature, relative humidity, precipitation and wind speed. Effects of individual weather parameters on hourly and daily concentrations of different fungal spore types were examined using Spearman's rank association test, whereas effects of complex of meteorological factors on hourly and daily compositions of spore were assessed using detrended correspondence analysis (DCA) and redundancy analysis (RDA). Airborne fungal spore distribution patterns in relation to meteorological variables were determined by RDA, after DCA results detected a linear structure of the spore data. The RDA results obtained indicated that all the applied variables accounted for 20 and 22% of the total variance in the hourly and daily spore data, respectively. The results of stepwise forward selection of variables revealed all included hourly and daily meteorological variables were statistically significant. The largest amount of the total variance in the spore composition was explained by the air temperature in both cases (16%). Multivariate ordination did not show large differences between the hourly and daily relationships (with exception of wind speed impact), while the differences between simple hourly and daily correlations were more clear. Correlations between daily values of variables were in most cases higher than between hourly values of variables.

Contribution of Leptosphaeria species ascospores to autumn asthma in areas of oilseed rape production.

BACKGROUND: An increase in the number of hospital admissions from September to November in the northern hemisphere has been frequently reported. At this time, some species of fungal genus Leptosphaeria produce numerous ascospores, which are easily airborne. However, we lack knowledge about whether Leptosphaeria produces allergenic proteins. OBJECTIVE: To evaluate the potential of Leptosphaeria ascospores to contribute to autumn asthma. METHODS: Detailed bioinformatic analysis of proteins produced by Leptosphaeria maculans available in databases was performed and the data compared with allergens found in other airborne fungi. The concentrations of Leptosphaeria ascospores detected at 2 sites were compared to these obtained in other environments worldwide. RESULTS: We found that Leptosphaeria species produce proteins with a high identity to commonly known aeroallergens of several well-characterized molds. The level of amino acid identity significantly exceeded the allergen identity thresholds recommended by the Food and Agricultural Organization/World Health Organization (35%), which indicates allergenic properties of L maculans and ensures the same properties in the other Leptosphaeria species. CONCLUSION: High concentrations of Leptosphaeria species ascospores in the autumn and postulated allergenicity of their proteins strongly suggest that this genus contributes to worldwide reported autumn asthma. The finding opens the question of allergenicity of the other never studied fungal species present in aeroplankton.

Madeira-a tourist destination for asthma sufferers.

Madeira Island is a famous tourist destination due to its natural and climatic values. Taking into account optimal weather conditions, flora richness and access to various substrates facilitating fungal growth, we hypothesised a very high risk of elevated fungal spore and pollen grain concentrations in the air of Funchal, the capital of Madeira. Concentration levels of the most allergenic taxa were measured from 2003 to 2009, using a 7-day volumetric air sampler, followed by microscopy analysis. Dependence of bioaerosols on the weather conditions and land use were assessed using spatial and statistical tools. Obtained results were re-visited by a comparison with hospital admission data recorded at the Dr. Nélio Mendonça Hospital in Funchal. Our results showed that despite propitious climatic conditions, overall pollen grain and fungal spore concentrations in the air were very low and did not exceed any clinically established threshold values. Pollen and spore peak concentrations also did not match with asthma outbreaks in the winter. Identification of places that are "free" from biological air pollution over the summer, such as Madeira Island, is very important from the allergic point of view.

A 10-year study of Alternaria and Cladosporium in two Polish cities (Szczecin and Cracow) and relationship with the meteorological parameters.

Alternaria and Cladosporium spores belong to the most frequent and allergenic particles in bioaerosol in the temperate climate. The investigation of Alternaria and Cladosporium spore concentrations was performed in two cities in Poland, Szczecin and Cracow, in 2004-2013. The meteorological parameters taken to assess their impact on fungal spores were average, maximum and minimum temperature, relative humidity and average wind velocity. In order to reveal whether changes in dynamics of spore seasons are driven by meteorological conditions, ordination methods were applied. Canonical correspondence analysis was used to explore redundancy among the predictors (meteorological parameters). Prior to ordination analyses, the data were log(x)-transformed. Concentrations of Alternaria and Cladosporium spores were significantly higher in Szczecin comparing to Cracow, but it was also observed the decreasing trend in the spore concentrations in Szczecin. As regards temperature, it was higher in Cracow and was still increasing in the studied years. Relative humidity and wind velocity were significantly lower in Cracow. In Szczecin meteorological conditions did not explain changes in spore season characteristics (insignificant redundancy analysis models), while in Cracow's redundancy analysis models indicated that spore season parameters were in over 40 % determined by meteorological conditions, mainly air temperature and wind velocity. If they increase, the peak value, total number of spores and their average concentrations in a season will also increase.

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.

Effects of meteorological factors on the composition of selected fungal spores in the air.

The aim of the study was to determine functional relationships between composition of air spora and meteorological factors, using multivariate statistical technique: canonical correspondence analysis (CCA). Analyses were conducted for the data collected during the 4 year (2007-2010) and, in order to show the dynamics of such relationships, for each year separately. The CCA results indicated that all statistically significant variables accounted for 15.3 % of the total variance in the spore data in the 4 years. The largest amount of the total variance was explained in this period by the mean air temperature (9.2 %). The meteorological factors impacted spore composition differently in different years, when analysis was done for each year separately. The highest values of the total variance in the spore data, explained by the statistically significant variables, were found in 2010 (32.3 %), with the highest contribution of mean air temperature (23.8 %). In that year, the above-mentioned parameter had the lowest value in comparison to other years. Canonical correspondence analysis provides not only a comprehensive assessment of the impact of meteorological factors on specific spore combinations in the air, but also informative graphical presentations of the results, illustrating the correlation between the occurrence of particular spore taxa and meteorological variables.

Changes in concentration of Alternaria and Cladosporium spores during summer storms.

Fungal spores are known to cause allergic sensitization. Recent studies reported a strong association between asthma symptoms and thunderstorms that could be explained by an increase in airborne fungal spore concentrations. Just before and during thunderstorms the values of meteorological parameters rapidly change. Therefore, the goal of this study was to create a predictive model for hourly concentrations of atmospheric Alternaria and Cladosporium spores on days with summer storms in Szczecin (Poland) based on meteorological conditions. For this study we have chosen all days of June, July and August (2004-2009) with convective thunderstorms. There were statistically significant relationships between spore concentration and meteorological parameters: positive for air temperature and ozone content while negative for relative humidity. In general, before a thunderstorm, air temperature and ozone concentration increased, which was accompanied by a considerable increase in spore concentration. During and after a storm, relative humidity increased while both air temperature ozone concentration along with spore concentrations decreased. Artificial neural networks (ANN) were used to assess forecasting possibilities. Good performance of ANN models in this study suggest that it is possible to predict spore concentrations from meteorological variables 2 h in advance and, thus, warn people with spore-related asthma symptoms about the increasing abundance of airborne fungi on days with storms.

Co-exposure to highly allergenic airborne pollen and fungal spores in Europe.

The study is aimed at determining the potential spatiotemporal risk of the co-occurrence of airborne pollen and fungal spores high concentrations in different bio-climatic zones in Europe. Birch, grass, mugwort, ragweed, olive pollen and Alternaria and Cladosporium fungal spores were investigated at 16 sites in Europe, in 2005-2019. In Central and northern Europe, pollen and fungal spore seasons mainly overlap in June and July, while in South Europe, the highest pollen concentrations occur frequently outside of the spore seasons. In the coldest climate, no allergy thresholds were exceeded simultaneously by two spore or pollen taxa, while in the warmest climate most of the days with at least two pollen taxa exceeding threshold values were observed. The annual air temperature amplitude seems to be the main bioclimatic factor influencing the accumulation of days in which Alternaria and Cladosporium spores simultaneously exceed allergy thresholds. The phenomenon of co-occurrence of airborne allergen concentrations gets increasingly common in Europe and is proposed to be present on other continents, especially in temperate climate.

Occurrence of Didymella ascospores in western and southern Poland in 2004-2006.

The concentration of airborne Didymella spores has been investigated at two monitoring sites situated along the west-south transect in Poland (Szczecin, Kraków), i.e. from a height of 100 to 219 m, respectively, above sea level. The aerobiological monitoring of fungal spores was performed by means of two Lanzoni volumetric spore traps. The high Didymella spore numbers were observed at both cities in June, July and August. Statistically significant correlations have been found mainly between the Didymella spore concentrations in the air and the minimum air temperature and relative air humidity. The spore count of Didymella is determined by the diversity of local flora and weather conditions, especially by the relative air humidity. The identification of factors that influence and shape spore concentrations may significantly improve the current methods of allergy prevention.

Bioaerosols on the atmospheric super highway: An example of long distance transport of Alternaria spores from the Pannonian Plain to Poland.

Alternaria spores are pathogenic to agricultural crops, and the longest and the most severe sporulation seasons are predominantly recorded in rural areas, e.g. the Pannonian Plain (PP) in South-Central Europe. In Poland (Central Europe), airborne Alternaria spore concentrations peak between July and August. In this study, we test the hypothesis that the PP is the source of Alternaria spores recorded in Poland after the main sporulation season (September-October). Airborne Alternaria spores (2005-2019) were collected using volumetric Hirst spore traps located in 38 locations along the potential pathways of air masses, i.e. from Serbia, Romania and Hungary, through the Czech Republic, Slovakia and Ukraine, to Northern Poland. Three potential episodes of Long Distance Transport (LDT) were selected and characterized in detail, including the analysis of Alternaria spore data, back trajectory analysis, dispersal modelling, and description of local weather and mesoscale synoptic conditions. During selected episodes, increases in Alternaria spore concentrations in Poznan were recorded at unusual times that deviated from the typical diurnal pattern, i.e. at night or during morning hours. Alternaria spore concentrations on the PP were very high (>1000 spores/m3) at that time. The presence of non-local Ambrosia pollen, common to the PP, were also observed in the air. Air mass trajectory analysis and dispersal modelling showed that the northwest part of the PP, north of the Transdanubian Mountains, was the potential source area of Alternaria spores. Our results show that Alternaria spores are transported over long distances from the PP to Poland. These spores may markedly increase local exposure to Alternaria spores in the receptor area and pose a risk to both human and plant health. Alternaria spores followed the same atmospheric route as previously described LDT ragweed pollen, revealing the existence of an atmospheric super highway that transports bioaerosols from the south to the north of Europe.

The effects of meteorological factors on the occurrence of Ganoderma sp. spores in the air.

Ganoderma sp. is an airborne fungal spore type known to trigger respiratory allergy symptoms in sensitive patients. Aiming to reduce the risk for allergic individuals, we analysed fungal spore circulation in Szczecin, Poland, and its dependence on meteorological conditions. Statistical models for the airborne spore concentrations of Ganoderma sp.-one of the most abundant fungal taxa in the area-were developed. Aerobiological sampling was conducted over 2004-2008 using a volumetric Lanzoni trap. Simultaneously, the following meteorological parameters were recorded: daily level of precipitation, maximum and average wind speed, relative humidity and maximum, minimum, average and dew point temperatures. These data were used as the explaining variables. Due to the non-linearity and non-normality of the data set, the applied modelling techniques were artificial neural networks (ANN) and mutlivariate regression trees (MRT). The obtained classification and MRT models predicted threshold conditions above which Ganoderma sp. appeared in the air. It turned out that dew point temperature was the main factor influencing the presence or absence of Ganoderma sp. spores. Further analysis of spore seasons revealed that the airborne fungal spore concentration depended only slightly on meteorological factors.

Airborne Aspergillus and Penicillium in the atmosphere of Szczecin, (Poland) (2004-2009).

The investigation into airborne fungal spore concentrations was conducted in Szczecin (Poland) between 2004 and 2009. The objective of the studies was to determine a seasonal variation in concentrations of amerospores on the basis of meteorological parameters. The presence of spores in Szczecin was recorded using a volumetric method. Fungal spores were present in the air in high numbers in late summer and early autumn. The highest concentrations were noted in September, October and November. The peak period was recorded in August, September, October and November. The highest annual number of spores occurred in 2005 and 2007 and the lowest in 2006. High values of daily concentration of amerospores occurred during the afternoon and late at night. In 2005 and 2007 the late-night maximum was overdue about 1 or 2 h. For daily values of dew point temperature and relative humidity, the coefficients were positive, significant for p = 0.001 and ranged from 0.342 to 0.258. The average wind speed was positively correlated for p = 0.01 and the coefficient was 0.291. The similar relations were noted for hourly values of spore concentrations for p = 0.05, p = 0.01 and p = 0.001. For these spore types, the dew point temperature and relative humidity appeared to be the most influential factor.

Abundance of Ganoderma sp. in Europe and SW Asia: modelling the pathogen infection levels in local trees using the proxy of airborne fungal spore concentrations.

Ganoderma comprises a common bracket fungal genus that causes basal stem rot in deciduous and coniferous trees and palms, thus having a large economic impact on forestry production. We estimated pathogen abundance using long-term, daily spore concentration data collected in five biogeographic regions in Europe and SW Asia. We hypothesized that pathogen abundance in the air depends on the density of potential hosts (trees) in the surrounding area, and that its spores originate locally. We tested this hypothesis by (1) calculating tree cover density, (2) assessing the impact of local meteorological variables on spore concentration, (3) computing back trajectories, (4) developing random forest models predicting daily spore concentration. The area covered by trees was calculated based on Tree Density Datasets within a 30 km radius from sampling sites. Variations in daily and seasonal spore concentrations were cross-examined between sites using a selection of statistical tools including HYSPLIT and random forest models. Our results showed that spore concentrations were higher in Northern and Central Europe than in South Europe and SW Asia. High and unusually high spore concentrations (> 90th and > 98th percentile, respectively) were partially associated with long distance transported spores: at least 33% of Ganoderma spores recorded in Madeira during days with high concentrations originated from the Iberian Peninsula located >900 km away. Random forest models developed on local meteorological data performed better in sites where the contribution of long distance transported spores was lower. We found that high concentrations were recorded in sites with low host density (Leicester, Worcester), and low concentrations in Kastamonu with high host density. This suggests that south European and SW Asian forests may be less severely affected by Ganoderma. This study highlights the effectiveness of monitoring airborne Ganoderma spore concentrations as a tool for assessing local Ganoderma pathogen infection levels.

Hourly predictive artificial neural network and multivariate regression tree models of Alternaria and Cladosporium spore concentrations in Szczecin (Poland).

A study was made of the link between time of day, weather variables and the hourly content of certain fungal spores in the atmosphere of the city of Szczecin, Poland, in 2004-2007. Sampling was carried out with a Lanzoni 7-day-recording spore trap. The spores analysed belonged to the taxa Alternaria and Cladosporium. These spores were selected both for their allergenic capacity and for their high level presence in the atmosphere, particularly during summer. Spearman correlation coefficients between spore concentrations, meteorological parameters and time of day showed different indices depending on the taxon being analysed. Relative humidity (RH), air temperature, air pressure and clouds most strongly and significantly influenced the concentration of Alternaria spores. Cladosporium spores correlated less strongly and significantly than Alternaria. Multivariate regression tree analysis revealed that, at air pressures lower than 1,011 hPa the concentration of Alternaria spores was low. Under higher air pressure spore concentrations were higher, particularly when RH was lower than 36.5%. In the case of Cladosporium, under higher air pressure (>1,008 hPa), the spores analysed were more abundant, particularly after 0330 hours. In artificial neural networks, RH, air pressure and air temperature were the most important variables in the model for Alternaria spore concentration. For Cladosporium, clouds, time of day, air pressure, wind speed and dew point temperature were highly significant factors influencing spore concentration. The maximum abundance of Cladosporium spores in air fell between 1200 and 1700 hours.

Airborne Alternaria and Cladosporium fungal spores in Europe: Forecasting possibilities and relationships with meteorological parameters.

Airborne fungal spores are prevalent components of bioaerosols with a large impact on ecology, economy and health. Their major socioeconomic effects could be reduced by accurate and timely prediction of airborne spore concentrations. The main aim of this study was to create and evaluate models of Alternaria and Cladosporium spore concentrations based on data on a continental scale. Additional goals included assessment of the level of generalization of the models spatially and description of the main meteorological factors influencing fungal spore concentrations. Aerobiological monitoring was carried out at 18 sites in six countries across Europe over 3 to 21 years depending on site. Quantile random forest modelling was used to predict spore concentrations. Generalization of the Alternaria and Cladosporium models was tested using (i) one model for all the sites, (ii) models for groups of sites, and (iii) models for individual sites. The study revealed the possibility of reliable prediction of fungal spore levels using gridded meteorological data. The classification models also showed the capacity for providing larger scale predictions of fungal spore concentrations. Regression models were distinctly less accurate than classification models due to several factors, including measurement errors and distinct day-to-day changes of concentrations. Temperature and vapour pressure proved to be the most important variables in the regression and classification models of Alternaria and Cladosporium spore concentrations. Accurate and operational daily-scale predictive models of bioaerosol abundances contribute to the assessment and evaluation of relevant exposure and consequently more timely and efficient management of phytopathogenic and of human allergic diseases.

Detection of soft wheat in semolina and durum wheat bread by analysis of DNA microsatellites.

The aim of this work was to evaluate the analysis of DNA microsatellites for the detection of soft wheat (Triticum aestivum L.) in semolina and durum wheat bread (prepared from Triticum turgidum L. var. durum). The results enabled selection of an efficient D-genome-specific repetitive DNA sequence to detect common wheat in semolina and breads by qualitative PCR with a threshold of 3 and 5%, respectively, lowered to 2.5% by real-time PCR. This is of major importance for checking during production of some typical products recently awarded the European Protected Designation of Origin (PDO) mark such as Altamura bread, which should not contain soft wheat flour. The feasibility of quantification of common wheat adulteration in semolina using real-time PCR was also demonstrated.

Back-trajectory modelling and DNA-based species-specific detection methods allow tracking of fungal spore transport in air masses.

Recent advances in molecular detection of living organisms facilitate the introduction of novel methods to studies of the transport of fungal spores over large distances. Monitoring the migration of airborne fungi using microscope based spore identification is limited when different species produce very similar spores. In our study, DNA-based monitoring with the use of species-specific probes allowed us to track the aerial movements of two important fungal pathogens of oilseed rape (Brassica napus L.), i.e., Leptosphaeria maculans and Leptosphaeria biglobosa, which have identical spore shape and size. The fungi were identified using dual-labelled fluorescent probes that were targeted to a ß-tubulin gene fragment of either Leptosphaeria species. Spore identification by Real-Time PCR techniques capable of detecting minute amounts of DNA of selected fungal species was combined with back-trajectory analysis, allowing the tracking of past movements of air masses using the Hybrid Single Particle Lagrangian Integrated Trajectory model. Over a study period spanning the previous decade (2006-2015) we investigated two specific events relating to the long distance transport of Leptosphaeria spp. spores to Szczecin in North-West Poland. Based on the above mentioned methods and the results obtained with the additional spore sampler located in nearby Szczecin, and operating at the ground level in an oilseed rape field, we have demonstrated that on both occasions the L. biglobosa spores originated from the Jutland Peninsula. This is the first successful attempt to combine analysis of back-trajectories of air masses with DNA-based identification of economically important pathogens of oilseed rape in Europe. In our studies, the timing of L. biglobosa ascospore dispersal in the air was unlikely to result in the infection of winter oilseed rape grown as a crop plant. However, the fungus could infect other host plants, such as vegetable brassicas, cruciferous weeds, spring rapeseed and winter rapeseed growing as a volunteer plant.

Seasonal variation of Ganoderma spore concentrations in urban and suburban districts of the city of Szczecin, Poland.

According to recent studies, Ganoderma may be the third genus, after Alternaria and Cladosporium, the spores of which cause symptoms of allergy, and concentration is related to meteorological factors. The aerobiology of Ganoderma spores in Szczecin in urban and suburban districts was examined using Lanzoni Volumetric Spore Traps in 2008-2010. Ganoderma spores were present in the atmosphere on more than 90% of the days from June through September with peak concentrations in June, July and September. The number of days with spores was lower in the suburban district, while the total number of spores collected was higher there than in the urban district. Correlation and multiple regression analyses revealed weak relationships between Ganoderma and meteorological conditions, while testing the significance of differences between the districts showed that urban development did not have a clear impact on the values of meteorological parameters. A significantly higher abundance of spores in the suburbs of Szczecin seemed to be conditioned by the closeness of potential area sources. This study indicates that a single measuring site in the city centre insufficiently reflected the dynamics and level of Ganoderma spore concentration in peripheral districts.