Bioaerosols are useful indicators of plant phenology and can demonstrate the impacts of climate change on both local and regional scales (e.g. pollen monitoring/flowering phenology). Analysing bioaerosols with eDNA approaches are becoming more popular to quantify the diversity of airborne plant environmental DNA (eDNA) and flowering season of plants and trees. Leaf abscission from broadleaved trees and other perennial species can also indicate the status of plant health in response to climate. This happens primarily during autumn in response to seasonal growth conditions and environmental factors, such as changing photoperiod and reduced temperatures. During this period biological material is released in larger quantities to the environment. Here, rural bioaerosol composition during late summer and autumn was captured by MiSEQ sequencing of the rRNA internal transcribed spacer 2 (ITS2) region, a common marker for taxonomic variation. Meteorological parameters were recorded from a proximal weather station. The composition of atmospheric taxa demonstrated that deciduous tree DNA forms part of the bioaerosol community during autumn and, for several common broadleaved tree species, atmospheric DNA abundance correlated to high wind events. This suggests that both flowering and autumn storms cause bioaerosols from deciduous trees that can be detected with eDNA approaches. This is an aspect that must be considered when eDNA methods are used to analyse either pollen or other fragments from trees.
Alternaria is a ubiquitous fungal genus with many allergenic and pathogenic species inhabiting grasslands. We hypothesise that grasslands (natural/man-made) host a diversity of fungal species whose spores have varying emission patterns. Therefore, the purpose of this study was to examine the potential of grasslands for emission, diversity and composition of Alternaria and other fungal species. To test the hypothesis, Hirst-type and multi-vial Cyclone samplers collected air samples from two grassland sites (unmanaged and managed) and a non-grassland site at Lakeside campus of the University of Worcester, United Kingdom for the period May to September 2019. The unmanaged grassland was originally planted with grasses and left uncut for three years. The managed grassland was a roadside verge that was cut once every year, typically after most grasses have flowered. We used optical microscopy and Illumina MiSeq sequencing to investigate the emission, abundance, diversity and composition of the fungal spores from each site alongside meteorological variables. Kruskal-Wallis and Wilcoxon tests examined differences in the bi-hourly Alternaria concentrations between the sites. Shannon's and Simpson's Index determined the diversity of the fungal spores between the unmanaged and non-grassland sites. The results showed that grasslands are a strong source of Alternaria spores with considerably higher numbers of clinically important days compared with the non-grassland site. The managed grassland varied in Alternaria spore emission pattern from the unmanaged, probably due to differences in environmental variables and cutting frequency. The unmanaged grassland and non-grassland sites showed a high diversity of fungi including Alternaria, Cladosporium, Ascochyta, Botrytis and Aureobasidium. Overall, the study shows that grasslands are a strong source of fungal spores with allergenic and pathogenic potential and have varying emission patterns, compared with nearby urban areas where monitoring stations are located. This information is useful for atmospheric modelling of airborne fungal spore sources and has implications for allergy sufferers in particular.
This study examines the influence of meteorological factors and air pollutants on the performance of automatic pollen monitoring devices, as part of the EUMETNET Autopollen COST ADOPT-intercomparison campaign held in Munich, Germany, during the 2021 pollen season. The campaign offered a unique opportunity to compare all automatic monitors available at the time, a Plair Rapid-E, a Hund-Wetzlar BAA500, an OPC Alphasense, a KH-3000 Yamatronics, three Swisens Polenos, a PollenSense APS, a FLIR IBAC2, a DMT WIBS-5, an Aerotape Sextant, to the average of four manual Hirst traps, under the same environmental conditions. The investigation aimed to elucidate how meteorological factors and air pollution impact particle capture and identification efficiency.The analysis showed coherent results for most devices regarding the correlation between environmental conditions and pollen concentrations. This reflects on one hand, a significant correlation between weather and airborne pollen concentration, and on the other hand the capability of devices to provide meaningful data under the conditions under which measurements were taken. However, correlation strength varied among devices, reflecting differences in design, algorithms, or sensors used. Additionally, it was observed that different algorithms applied to the same dataset resulted in different concentration outputs, highlighting the role of algorithm design in these systems (monitor + algorithm).Notably, no significant influence from air pollutants on the pollen concentrations was observed, suggesting that any potential difference in effect on the systems might require higher air pollution concentrations or more complex interactions. However, results from some monitors were affected to a minor degree by specific weather variables.Our findings suggest that the application of real-time devices in urban environments should focus on the associated algorithm that classifies pollen taxa. The impact of air pollution, although not to be excluded, is of secondary concern as long as the pollution levels are similar to a large European city like Munich.
The advent of automatic pollen and fungal spore monitoring over the past few years has brought about a paradigm change. The provision of real-time information at high temporal resolution opens the door to a wide range of improvements in terms of the products and services made available to a widening range of end-users and stakeholders. As technology and methods mature, it is essential to properly quantify the impact automatic monitoring has on the different end-user domains to better understand the real long-term benefits to society. In this paper, we focus the main domains where such impacts are expected, using Europe as a basis to provide qualitative estimates and to describe research needs to better quantify impacts in future. This will, in part, also serve to justify further investment and help to expand monitoring networks.
Fungi are among the most diverse and ecologically important kingdoms in life. However, the distributional ranges of fungi remain largely unknown as do the ecological mechanisms that shape their distributions(1,2). To provide an integrated view of the spatial and seasonal dynamics of fungi, we implemented a globally distributed standardized aerial sampling of fungal spores(3). The vast majority of operational taxonomic units were detected within only one climatic zone, and the spatiotemporal patterns of species richness and community composition were mostly explained by annual mean air temperature. Tropical regions hosted the highest fungal diversity except for lichenized, ericoid mycorrhizal and ectomycorrhizal fungi, which reached their peak diversity in temperate regions. The sensitivity in climatic responses was associated with phylogenetic relatedness, suggesting that large-scale distributions of some fungal groups are partially constrained by their ancestral niche. There was a strong phylogenetic signal in seasonal sensitivity, suggesting that some groups of fungi have retained their ancestral trait of sporulating for only a short period. Overall, our results show that the hyperdiverse kingdom of fungi follows globally highly predictable spatial and temporal dynamics, with seasonality in both species richness and community composition increasing with latitude. Our study reports patterns resembling those described for other major groups of organisms, thus making a major contribution to the long-standing debate on whether organisms with a microbial lifestyle follow the global biodiversity paradigms known for macroorganisms(4,5).
Novel methods for sampling and characterizing biodiversity hold great promise for re-evaluating patterns of life across the planet. The sampling of airborne spores with a cyclone sampler, and the sequencing of their DNA, have been suggested as an efficient and well-calibrated tool for surveying fungal diversity across various environments. Here we present data originating from the Global Spore Sampling Project, comprising 2,768 samples collected during two years at 47 outdoor locations across the world. Each sample represents fungal DNA extracted from 24 m3 of air. We applied a conservative bioinformatics pipeline that filtered out sequences that did not show strong evidence of representing a fungal species. The pipeline yielded 27,954 species-level operational taxonomic units (OTUs). Each OTU is accompanied by a probabilistic taxonomic classification, validated through comparison with expert evaluations. To examine the potential of the data for ecological analyses, we partitioned the variation in species distributions into spatial and seasonal components, showing a strong effect of the annual mean temperature on community composition.
This research builds upon a previous study that explored the potential of the modified WIBS-4+ to selectively differentiate and detect different bioaerosol classes. The current work evaluates the influence of meteorological and air quality parameters on bioaerosol concentrations, specifically pollen and fungal spore dynamics.Temperature was found to be the most influential parameter in terms of pollen production and release, showing a strong positive correlation. Wind data analysis provided insights into the potential geographic origins of pollen and fungal spore concentrations. Fungal spores were primarily shown to originate from a westerly direction, corresponding to agricultural land use, whereas pollen largely originated from a North-easterly direction, corresponding to several forests.The influence of air quality was also analysed to understand its potential impact on the WIBS fluorescent parameters investigated. Most parameters had a negative association with fungal spore concentrations, whereas several anthropogenic influences showed notable positive correlations with daily pollen concentrations. This is attributed to similar driving forces (meteorological parameters) and geographical origins. In addition, the WIBS showed a significant correlation with anthropogenic pollutants originating from combustion sources, suggesting the potential for such modified spectroscopic instruments to be utilized as air quality monitors.By combining all meteorological and pollution data along with WIBS-4+ channel data, a set of Multiple Linear Regression (MLR) analyses were completed. Successful results with R2 values ranging from 0.6 to 0.8 were recorded. The inclusion of meteorological parameters was dependent on the spore or pollen type being examined.
Aeroallergens or inhalant allergens, are proteins dispersed through the air and have the potential to induce allergic conditions such as rhinitis, conjunctivitis, and asthma. Outdoor aeroallergens are found predominantly in pollen grains and fungal spores, which are allergen carriers. Aeroallergens from pollen and fungi have seasonal emission patterns that correlate with plant pollination and fungal sporulation and are strongly associated with atmospheric weather conditions. They are released when allergen carriers come in contact with the respiratory system, e.g. the nasal mucosa. In addition, due to the rupture of allergen carriers, airborne allergen molecules may be released directly into the air in the form of micronic and submicronic particles (cytoplasmic debris, cell wall fragments, droplets etc.) or adhered onto other airborne particulate matter. Therefore, aeroallergen detection strategies must consider, in addition to the allergen carriers, the allergen molecules themselves. This review article aims to present the current knowledge on inhalant allergens in the outdoor environment, their structure, localization, and factors affecting their production, transformation, release or degradation. In addition, methods for collecting and quantifying aeroallergens are listed and thoroughly discussed. Finally, the knowledge gaps, challenges and implications associated with aeroallergen analysis are described.
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.
To benefit allergy patients and the medical practitioners, pollen information should be available in both a reliable and timely manner; the latter is only recently possible due to automatic monitoring. To evaluate the performance of all currently available automatic instruments, an international intercomparison campaign was jointly organised by the EUMETNET AutoPollen Programme and the ADOPT COST Action in Munich, Germany (March-July 2021). The automatic systems (hardware plus identification algorithms) were compared with manual Hirst-type traps. Measurements were aggregated into 3-hourly or daily values to allow comparison across all devices. We report results for total pollen as well as for Betula, Fraxinus, Poaceae, and Quercus, for all instruments that provided these data. The results for daily averages compared better with Hirst observations than the 3-hourly values. For total pollen, there was a considerable spread among systems, with some reaching R2 > 0.6 (3 h) and R2 > 0.75 (daily) compared with Hirst-type traps, whilst other systems were not suitable to sample total pollen efficiently (R2 < 0.3). For individual pollen types, results similar to the Hirst were frequently shown by a small group of systems. For Betula, almost all systems performed well (R2 > 0.75 for 9 systems for 3-hourly data). Results for Fraxinus and Quercus were not as good for most systems, while for Poaceae (with some exceptions), the performance was weakest. For all pollen types and for most measurement systems, false positive classifications were observed outside of the main pollen season. Different algorithms applied to the same device also showed different results, highlighting the importance of this aspect of the measurement system. Overall, given the 30 % error on daily concentrations that is currently accepted for Hirst-type traps, several automatic systems are currently capable of being used operationally to provide real-time observations at high temporal resolutions. They provide distinct advantages compared to the manual Hirst-type measurements.
Characterizing pollen release and dispersion processes is fundamental for knowledge advancement in ecological, agricultural and public health disciplines. Understanding pollen dispersion from grass communities is especially relevant due to their high species-specific allergenicity and heterogeneously distributed source areas. Here, we aimed to address questions concerning fine level heterogeneity in grass pollen release and dispersion processes, with a focus on characterizing the taxonomic composition of airborne grass pollen over the grass flowering season using eDNA and molecular ecology methods. High resolution grass pollen concentrations were compared between three microscale sites (<300 m apart) in a rural area in Worcestershire, UK. The grass pollen was modelled with local meteorology in a MANOVA (Multivariate ANOVA) approach to investigate factors relevant to pollen release and dispersion. Simultaneously, airborne pollen was sequenced using Illumina MySeq for metabarcoding, analysed against a reference database with all UK grasses using the R packages DADA2 and phyloseq to calculate Shannon's Diversity Index (α-diversity). The flowering phenology of a local Festuca rubra population was observed. We found that grass pollen concentrations varied on a microscale level, likely attributed to local topography and the dispersion distance of pollen from flowering grasses in local source areas. Six genera (Agrostis, Alopecurus, Arrhenatherum, Holcus, Lolium and Poa) dominated the pollen season, comprising on average 77 % of the relative abundance of grass species reads. Temperature, solar radiation, relative humidity, turbulence and wind speeds were found to be relevant for grass pollen release and dispersion processes. An isolated flowering Festuca rubra population contributed almost 40 % of the relative pollen abundance adjacent to the nearby sampler, but only contributed 1 % to samplers situated 300 m away. This suggests that most emitted grass pollen has limited dispersion distance and our results show substantial variation in airborne grass species composition over short geographical scales.
Alternaria is a plant pathogen and human allergen. Alternaria alternata is one of the most abundant fungal spores in the air. The purpose of this study was to examine whether Alternaria spp . spore concentrations can be used to predict the abundance and spatio-temporal pattern of A . alternata spores in the air. This was investigated by testing the hypothesis that A . alternata dominates airborne Alternaria spp. spores and varies spatio-temporally. Secondarily, we aimed at investigating the relationship between airborne Alternaria spp. spores and the DNA profile of A . alternata spores between two proximate (~ 7 km apart) sites. These were examined by sampling Alternaria spp. spores using Burkard 7-day and cyclone samplers for the period 2016–2018 at Worcester and Lakeside campuses of the University of Worcester, UK. Daily Alternaria spp. spores from the Burkard traps were identified using optical microscopy whilst A . alternata from the cyclone samples was detected and quantified using quantitative polymerase chain reaction (qPCR). The results showed that either A . alternata or other Alternaria species spores dominate the airborne Alternaria spore concentrations, generally depending on weather conditions. Furthermore, although Alternaria spp. spore concentrations were similar for the two proximate sites, A . alternata spore concentrations significantly varied for those sites and it is highly likely that the airborne samples contained large amounts of small fragments of A . alternata . Overall, the study shows that there is a higher abundance of airborne Alternaria allergen than reported by aerobiological networks and the majority is likely to be from spore and hyphal fragments.
Abundance and diversity of airborne pollen are important to human health and biodiversity. The UK operational network collects airborne pollen from 8 flowering trees, grasses and three weeds using Hirst traps and microscopic identification from urban areas. Knowledge of total pollen diversity and differences between rural and urban zones is limited. We collect environmental DNA (eDNA) from air during summer and autumn over 3 years with mini cyclones from one urban and one rural site. Data are analysed using next generation sequencing and metabarcoding. We find the most common genus, Urtica (57%), is also identified by the national network. The grasses Lolium (10%), Agrostis (2%) and Holcus (1%) are in the national network grouped at family level, while Brassica (2%), Chenopodium (1%), Impatiens (2%), Plantago (4%) and Tilia (7%) are not part of the UK operational network. DNA from 138 genera was identified, where 2% of the sample could not be associated with specific genera. 40% of the sample was classified better using eDNA methods at the genus level, than by optical methods. We calculate Bray-Curtis dissimilarity for the rural and urban zones and find a systematic difference in biodiversity. Overall, this shows airborne DNA reveals more information than methods based on morphological differences. The results also suggest data from sites located in large urban areas will be less representative for less populated rural areas. This presents a dilemma in balancing a network and the associated costs delivering health relevant information to the most populated areas vs. a nation-wide approach.
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 Poznań 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.
Alternaria is a human/animal allergen and plant/animal pathogen. Cereal harvesting emits a large amount of Alternaria spores into the atmosphere. However, estimating the peak spore periods and source areas from large areas is often a challenge because of insufficient observation stations. The purpose of this study was to examine, using remote sensing and an atmospheric transport and dispersion model, the contribution of cereal harvesting to peak Alternaria spore concentrations. Daily Alternaria spores were collected using Hirst-type traps in 12 sites in Europe for the period 2016-2018. Hybrid Single-Particle Lagrangian Integrated Trajectory (HYSPLIT) back -trajectory and dispersion model was integrated with Sentinel-2 satellite imagery, Corine Land Cover 2018 (CLC2018) and Eurostat cereal data 2016 to map the Alternaria spore peaks and source areas in the 12 sites. Ground truth harvest data, collected at Worcester, UK, in 2018, and meteorological data were used to determine any effect of cereal harvesting and weather on peak spore concentrations. The results showed that the Sentinel-2 satellite detected agricultural areas that underwent intensive harvesting and this coincided with a rapid increase of Alternaria spore concentrations. Furthermore, local agricultural areas cultivated with cereals were the main sources of the peak Alternaria spore concentrations in all the study sites. Remote agricultural and non-agricultural sources, to a lesser extent, contributed to the peak spore concentrations at some sites, e.g. Borstel, Leicester and Worcester. Temperature during the harvesting periods (July and August) was found to significantly contribute to the peak spore concentrations. Overall, the study showed that it is possible to use Sentinel-2 satellite data alongside atmospheric transport and dispersion models to estimate periods of peak Alternaria spore concentra-tions and sources at a continental scale. This approach can be replicated for other bioaerosols that affect human health, agriculture and forestry.
Identifying the origin of bioaerosols is of central importance in many biological disciplines, such as human health, agriculture, forestry, aerobiology and conservation. Modelling sources, transportation pathways and sinks can reveal how bioaerosols vary in the atmosphere and their environmental impact. Grass pollen are particularly important due to their widely distributed source areas, relatively high abundance in the atmosphere and high allergenicity. Currently, studies are uncertain regarding sampler representability between distance and sources for grass pollen. Using generalized linear modelling, this study aimed to analyse this relationship further by answering the question of distance-to-source area contribution. Grass pollen concentrations were compared between urban and rural locations, located 6.4 km apart, during two years in Worcestershire, UK. We isolated and refined vegetation areas at 100 m × 100 m using the 2017 CEH Crop Map and conducted atmospheric modelling using HYSPLIT to identify which source areas could contribute pollen. Pollen concentrations were then modelled with source areas and meteorology using generalized linear mixed-models with three temporal variables as random variation. We found that the Seasonal Pollen Integral for grass pollen varied between both years and location, with the urban location having higher levels. Day of year showed higher temporal variation than the diurnal or annual variables. For the urban location, grass source areas within 30 km had positive significant effects in predicting grass pollen concentrations, while source areas within 2-10 km were important for the rural one. The source area differential was likely influenced by an urban-rural gradient that caused differences in the source area contribution. Temperature had positive highly significant effects on both locations while precipitation affected only the rural location. Combining atmospheric modelling, vegetation source maps and generalized linear modelling was found to be a highly accurate tool to identify transportation pathways of bioaerosols in landscape environments.
Respiratory allergies triggered by pollen allergens represent a significant health concern to the Irish public. Up to now, Ireland has largely refrained from participating in long-term aerobiological studies. Recently, pollen monitoring has commenced in several sampling locations around Ireland. The first results of the pollen monitoring campaigns for Dublin (urban) and Carlow (rural) concerning the period 2017–2019 and 2018–2019, respectively, are presented herein. Additional unpublished pollen data from 1978–1980 and, 2010–2011 were also incorporated in creating the first pollen calendar for Dublin. During the monitoring period over 60 pollen types were identified with an average Annual Pollen Integral (APIn) of 32,217 Pollen × day/m3 for Dublin and 78,411 Pollen × day/m3 for Carlow. The most prevalent pollen types in Dublin were: Poaceae (32%), Urticaceae (29%), Cupressaceae/Taxaceae (11%), Betula (10%), Quercus (4%), Pinus (3%), Fraxinus (2%), Alnus (2%) and Platanus (1%). The predominant pollen types in Carlow were identified as Poaceae (70%), Urticaceae (12%), Betula (10%), Quercus (2%), Fraxinus (1%) and Pinus (1%). These prevalent pollen types increased in annual pollen concentration in both locations from 2018 to 2019 except for Fraxinus. Although higher pollen concentrations were observed for the Carlow (rural) site a greater variety of pollen types were identified for the Dublin (urban) site. The general annual trend in the pollen season began with the release of tree pollen in early spring, followed by the release of grass and herbaceous pollen which dominated the summer months with the annual pollen season coming to an end in October. This behaviour was illustrated for 21 different pollen types in the Dublin pollen calendar. The correlation between ambient pollen concentration and meteorological parameters was also examined and differed greatly depending on the location and study year. A striking feature was a substantial fraction of the recorded pollen sampled in Dublin did not correlate with the prevailing wind directions. However, using non-parametric wind regression, specific source regions could be determined such as Alnus originating from the Southeast, Betula originating from the East and Poaceae originating from the Southwest.
Pollen monitoring has traditionally been carried out using manual methods first developed in the early 1950s. Although this technique has been recently standardised, it suffers from several drawbacks, notably data usually only being available with a delay of 3–9 days and usually delivered at a daily resolution. Several automatic instruments have come on to the market over the past few years, with more new devices also under development. This paper provides a comprehensive overview of all available and developing automatic instruments, how they measure, how they identify airborne pollen, what impacts measurement quality, as well as what potential there is for further advancement in the field of bioaerosol monitoring.
Alternaria is a pathogenic and allergenic fungus affecting 400 plant species and 334 million people globally. This study aimed at assessing the diversity of Alternaria species in airborne samples collected from closely located (7 km apart) and heterogeneous sites (rural, urban and unmanaged grassland) in Worcester and Lakeside, the UK. A secondary objective was to examine how the ITS1 subregion varies from ITS2 in Alternaria species diversity and composition. Airborne spores were collected using Burkard 7-day and multi-vial Cyclone samplers for the period 5 July 2016–9 October 2019. Air samples from the Cyclone were amplified using the ITS1and ITS2 subregions and sequenced using Illumina MiSeq platform whereas those from the Burkard sampler were identified and quantified using optical microscopy. Optical microscopy and eDNA revealed a high abundance of Alternaria in the rural, urban and unmanaged sites. ITS1 and ITS2 detected five and seven different Alternaria species at the three sampling sites, respectively. A. dactylidicola , A. metachromatica and A. infectoria were the most abundant. The rural, urban and unmanaged grassland sites had similar diversity (PERMANOVA) of the species due to similarity in land use and proximity of the sites. Overall, the study showed that heterogeneous and neighbouring sites with similar land uses can have similar Alternaria species. It also demonstrated that an eDNA approach can complement the classical optical microscopy method in providing more precise information on fungal species diversity in an environment for targeted management. Similar studies can be replicated for other allergenic and pathogenic fungi.