Long distance pollen transport cause problems for determining the timing of birch pollen season in Fennoscandia by using phenological observations

The male flowering and leaf bud burst of birch take place almost simultaneously, suggesting that the observations of leaf bud burst could be used to determine the timing of birch pollen release. However, long‐distance transport of birch pollen before the onset of local flowering may complicate the utilization of phenological observations in pollen forecasting.

We compared the timing of leaf bud burst of silver birch with the timing of the stages of birch pollen season during an eight year period (1997–2004) at five sites in Finland. The stages of the birch pollen season were defined using four different thresholds: 1) the first date of the earliest three‐day period with airborne birch pollen counts exceeding 10 grains m^?3 air; and the dates when the accumulated pollen sum reaches 2) 5%; 3) 50% and 4) 95% of the annual total. Atmospheric modelling was used to determine the source areas for the observed long‐distance transported pollen, and the exploitability of phenological observations in pollen forecasting was evaluated.

Pair‐wise comparisons of means indicate that the timing of leaf bud burst fell closest to the date when the accumulated pollen sum reached 5% of the annual total, and did not differ significantly from it at any site (p<0.05; Student‐Newman‐Keuls test). It was found that the timing of leaf bud burst of silver birch overlaps with the first half of the main birch pollen season. However, phenological observations alone do not suffice to determine the timing of the main birch pollen season because of long‐distance transport of birch pollen.     

Numerical simulation of birch pollen dispersion with an operational weather forecast system

We included a parameterisation of the emissions of pollen grains into the comprehensive model system COSMO-ART. In addition, a detailed density distribution of birch trees within Switzerland was derived. Based on these new developments, we carried out numerical simulations of the dispersion of pollen grains for an episode that occurred in April 2006 over Switzerland and the adjacent regions. Since COSMO-ART is based on the operational forecast model of the German Weather Service, we are presenting a feasibility study of daily pollen forecast based on methods which have been developed during the last two decades for the treatment of anthropogenic aerosol. A comparison of the model results and very detailed pollen counts documents the current possibilities and the shortcomings of the method and gives hints for necessary improvements.     

Sources,impact and exchange of early-spring birch pollen in the Moscow region and Finland

The paper presents an assessment of birch pollen seasons in Finland and Russia. The re-analysis covered the period from 1994 to 2005 and was focused on suspected long-range transport events that were recorded both in Moscow and at several Finnish sites. In order to trace the origin of airborne pollen before the onset of local pollination, we used both aerobiological and phenological observations combined with forward and adjoint (inverse) dispersion model simulations. It is shown that, although the Moscow region is surrounded by extensive birch forests, it still receives substantial amounts of foreign pollen before local pollination. In the Moscow region, the sources of long-range-transported pollen are in the south and south-west, sometimes even in the east. In Finland, there are frequently cases, before the local flowering season, in which Finnish territory receives Russian pollen; however in the opposite direction, from Finland to the Moscow region, no transport episodes were unequivocally registered. Analysis of the end of the seasons was more problematic, due to contributions to pollen observations from local sources; this results in difficulties in the reliable identification of the long-range transport episodes. Apart from its short-term effects on the pollen seasons, long-range transport can have substantial impacts on the exchange of genetic material within Europe. A quick atmospheric pathway for gene transport can be important for adaptation of plants to a changing climate.     

Poaceae pollen in Galicia (N.W. Spain): characterisation and recent trends in atmospheric pollen season

Airborne Poaceae pollen counts are greatly influenced by weather-related parameters, but may also be governed by other factors. Poaceae pollen is responsible for most allergic reactions in the pollen-sensitive population of Galicia (Spain), and it is therefore essential to determine the risk posed by airborne pollen counts. The global climate change recorded over recent years may prompt changes in the atmospheric pollen season (APS). This survey used airborne Poaceae pollen data recorded for four Galician cities since 1993, in order to characterise the APS and note any trends in its onset, length and severity. Pollen sampling was performed using Hirst-type volumetric traps; data were subjected to Spearman’s correlation test and regression models, in order to detect possible correlations between different parameters and trends. The APS was calculated using ten different methods, in order to assess the influence of each on survey results. Finally, trends detected for the major weather-related parameters influencing pollen counts over the study period were compared with those recorded over the last 30 years. All four cities displayed a trend towards lower annual total Poaceae pollen counts, lower peak values and a smaller number of days on which counts exceeded 30, 50 and 100 pollen grains/m³. Moreover, the survey noted a trend towards delayed onset and shorter duration of the APS, although differences were observed depending on the criteria used to define the first and the last day of the APS.     

Structural study of biologically significant ligands with major birch pollen allergen Betv1 by docking and molecular dynamics simulation

The major birch pollen allergen, Betv1 of Betula verrucosa is the main causative agent of birch pollen allergy in humans. Betv1 is capable of binding several physiological ligands including fatty acids, flavones, cytokinins and sterols. Until now, no structural information from crystallography or NMR is available regarding binding mode of any of these ligands into the binding pocket of Betv1. In the present study thirteen ligands have been successfully docked into the hydrophobic cavity of Betv1 and binding free energies of the complexes have been calculated using AutoDock 3.0.5. A linear relationship with correlation coefficient (R2) of 0.6 is obtained between ΔG(b)s values plotted against their corresponding IC50 values. The complex formed between Betv1 and the best docking pose for each ligand has been optimized by molecular dynamics simulation. Here, we describe the ligand binding of Betv1, which provides insight into the biological function of this protein. This knowledge is required for structural alteration or inhibition of some of these ligands in order to modify the allergenic properties of this protein.     

Airborne birch pollen in Neuchâtel (Switzerland): onset, peak and daily patterns

An accurate forecast of the starting point of thebirch pollen season in Neuchâtel can be made byadding the positive daily average air temperature fromFebruary 1st onward until the figure 270 is reached.At this point, the birch trees are ready to bloom.After that, the daily average temperature has toexceed 10 °C to allow pollen release.Today, the birch pollen season starts some 19 daysearlier in the year than in the 1980's, a consequenceof a recent climate change.The daily patterns of airborne birch pollen isirregular. Moreover, pollen concentrations frequentlyexceed the threshold of the appearance of allergicsymptoms, except during rainfall. Therefore, the onlybehavioral recommendation that can be given to peopleallergic to birch pollen is to shorten as much aspossible the contact with outdoor air during the mainbirch pollen season.     

空气花粉变化规律和预测预报研究进展

空气花粉是一个地区大气环境评估的重要指标,随着人们生态环境保护意识以及自身健康意识的提高,该领域的研究已经成为综合生物学、环境科学、气象学和医学等学科的全球性的研究课题.不同地区受其地理位置、植物组成和气候等多种因素的影响,空气花粉种类和浓度的变化模式有所不同,一般情况,一个地区白天空气花粉浓度高,夜晚浓度低;春夏季花粉浓度高,冬季花粉浓度低;春夏季乔木类植物花粉浓度高,而秋季草本类植物花粉浓度高.空气花粉预测预报工作直接关系到一个地区农业、环境、医疗卫生以及民众生活等多方面,当前空气花粉的预测预报工作已经由原来的定性预测发展到当前通过统计学和数学方法建立数学模型进行定量化预测,不仅可以进行空气花粉种类和浓度的预测,还可以通过花粉的变化预测未来气候的变化,预测的精确度也比以往有很大提高.通过分析空气花粉研究的进展对未来该领域的研究工作进行了展望.     

Evaluation of indoor passive pollen sedimentation over 1 year: a possible source of contamination?

The study aimed to evaluate the possible contamination by passive indoor pollen sedimentation in a laboratory where routine pollen analyses were conducted, but where no particular laboratory policies were adopted to limit contamination. Gravimetric pollen deposition was observed on traps (petri dishes soaked with glycerol) set in the palynology laboratory, under an extractor hood and on the bench beside it over 1 year (1995–1996), and in an air filtered room in a flow cabinet and on its roof over 1 month for comparison. Under the extractor hood, three types of airborne particles were deposited: pollen grains, spores and algae, representing 32.35, 67.28 and 0.37%, respectively, of the total sedimented particles over 1 year. The number of pollen grains deposited on the surface trap ranged from 0 (27 November to 4 December 1995) to 707 (10–18 April 1995). The highest number of taxa during a weekly collection was 23 (9–15 May 1995). The pollen flora represented by anemophilous pollen (>90% of the trapped pollen) was related to the vegetation next to the laboratory:Acer, Carpinus, Castanea, Corylus, Cupressaceae, Pinus, Quercus, Salix, Taxus for trees and shrubs andArtemisia, Brassicaceae, Plantago, Poaceae, andUrticaceae for grasses and weeds. Indoor pollen deposition corresponded to the period of the outdoor pollination (macroscopic field observation) which lasted from March to the beginning of August. However, some pollen were almost always present in the collection (Poaceae, Salix, Castanea, Betula), reflecting the occurrence of pollen grains in the atmosphere out of the pollination period. Moreover, about five times more entomophilous pollen was found under the extractor hood compared to the other area of the laboratory; even in the flow cabinet of the air filtered room, 237 particles were captured (versus only 15 on the roof). These data suggest a possible human contamination during operations under the extractor hood or in the flow cabinet. Although few airborne pollen were found, possible contamination has to be considered in investigations where even low pollen quantities are of interest.     

Feasibility study on automated recognition of allergenic pollen: grass, birch and mugwort

Quantification of airborne pollen is an important tool in scientific research and patient care in allergy. The currently available method relies on microscopic examination of pollen slides, performed by qualified researchers. Although highly reliable, the method is labor intensive and requires extensive training of the researchers involved. In an approach to develop alternative detection methods, we performed a feasibility study on the automated recognition of the allergenic relevant pollen, grass, birch, and mugwort, by utilizing digital image analysis and pattern recognition tools. Of a total of 254 pollen samples (including 79 of grass, 79 of birch and 96 of mugwort), 97.2% were recognized correctly. This encouraging result provides a promising prospect for future developments.     

Viability of Cururbita pepo pollen: biophysical and structural data

During ageing of the short-lived pollen grains of Cucurbita pepo L., water loss was examined in relation to viability using biophysical (¹H-nuclear magnetic resonance, NMR) and cytological methods (fluorochromatic reaction test, freezefracture and scanning electron microscopy). A semi-logarithmic representation of the pollen weight loss demonstrated the complexity of the dehydration process. A the study of proton loss using ¹H-NMR indicated that two major releases water of had taken place, each with different flux rates. Pulse ¹H-NMR experiments showed the occurrene of non-exponential signal decay as a function of time, indicating the existence of different fractions of water in a pollen grain sample. These fractions leave the pollen grain at different times during pollen dehydration, and one of them (that of the so-called vital water) can be related to pollen viability. The quantity of protons giving a signal during pulse ¹H-NMR experiments was very low when the pollen grains were judged to be dead according to the fluorochromatic test. Freeze-fracture replicas of these dead pollen grains (less than 25% water content) showed that the plasma membrane had become detached from the intine surface; this ultrastructural feature might therefore be involved in the loss of pollen viability.Abbreviations A initial amplitude of the NMR signal - A₂ quantity of water charcterized by T_2-2 - A₅ quantity of water characterized by T_2–5 - FCR fluorochromatic reaction - NMR nuclear magnetic resonance - T₂ transverse relaxation time - T_2-2 T₂ measured with 2 ms between each pulse of radiofrequency - T_2–5 T₂ measured with 5 ms between each pulse of radiofrequency     

Antigenic cross-reactivity between Schistosoma mansoni and pollen allergens from the birch tree (Betula verrucosa) and Timothy grass (Phleum pratense): involvement of shared glycan epitopes and implications for the hygiene hypothesis

Previous studies have shown that schistosome infection can protect against allergic symptoms, but the underlying mechanisms are still not fully understood. Here we have shown that rabbit IgG antibodies raised against Schistosoma mansoni soluble egg antigens (SmSEA) are cross-reactive with a wide array of molecules in Timothy grass pollen (TGP) and birch tree pollen (BTP). Five of the cross-reactive pollen molecules (two from TGP and three from BTP) were selected randomly and identified by tandem mass spectrometric (TMS) analysis to be, respectively, the TGP allergens Phl p 1 and Phl p 5b, and BTP glutathione S-transferase (GST), and the BTP allergens Bet v 1 and Bet v 6.0102. Rabbit anti-SmSEA IgG antibodies that cross-reacted with each of the five allergens were found to be reactive with three major S. mansoni egg antigens, IPSE/alpha-1, omega-1 and kappa-5. Pairwise alignment of the amino acid sequences of each of the five TMS-identified pollen allergens with each of the three egg antigens revealed a low level of amino acid sequence identity. Further experiments indicated that the schistosome antigen/allergen cross-reactivity was mostly due to similar glycans present in helminths and plants, but not in mammals: so called cross-reactive carbohydrate determinants (CCDs). Previously, CCDs have been implicated in the cross-reactivity between many plants and invertebrates. Furthermore, pollen-induced anti-CCD IgGs have been found in sera of patients undergoing allergen-specific immunotherapy (SIT) and implicated in the treatment of the allergy. Thus, our finding provides not only possible explanations for the allergy-protective effect of helminth/schistosome infections as explained by the hygiene hypothesis, but also a potential starting point for improved SIT.     

Concentration of airborne pollen of Vitisvinifera L. and yield forecast: a case study at S.Michele all'Adige,Trento, Italy

The possibility of creating a forecast model for grapeproduction using airborne pollen concentrations ofVitis vinifera L. was investigated. The studyarea is located in the province of Trento (46°11,11°08), North Italy. Because of its prestigious wineproduction, the area is quite important on a nationalscale.Airborne pollen concentration data (P/m³) werecollected for a five year period (1993–1997) with aHirst type sampler (VPPS 2000 – Lanzoni) following thestandard methodology proposed by the ItalianAeroallergen Network. For the same five-year period,grape production data (kg/ha) from vineyards adjacentto the pollen sampler were used along with provincialdata. Regression lines were constructed using the Lnof the annual sum of daily pollen concentrations (Ip)as the independent variable and grape productionvalues as the dependent variable. Correlations betweenpollen data (pollen index, beginning and duration ofmain pollen season) and meteorological data(temperature and rainfall) are shown.     

Aerobiology of Artemisia airborne pollen in Murcia (SE Spain) and its relationship with weather variables: annual and intradiurnal variations for three different species. Wind vectors as a tool in determining pollen origin

Detailed results from a 2-year survey of airborne pollen concentrations of Artemisia in Murcia are presented. Three consecutive pollen seasons of Artemisia occurring each year, related to three different species (A. campestris, A. herba-alba and A. barrelieri), were observed. A winter blooming of Artemisia could explain the incidence of subsequent pollinosis in the Murcia area. With regard to meteorological parameters, mathematical analyses showed relationships between daily pollen concentrations of Artemisia in summer–autumn and precipitations that occurred 6–8 weeks before. The cumulative percentage of insolation from 1 March seemed to be related to blooming onsets. Once pollination has begun, meteorological factors do not seem to influence pollen concentrations significantly. Intradiurnal patterns of pollen concentrations were similar for late summer and winter species (A. campestris and A. barrelieri). During autumn blooming (A. herba-alba), the intradiurnal pattern was particularly erratic. Theoretical values of wind run were obtained for each pollen season by the graphical sum of hourly wind vectors. When theoretical wind run was mapped onto the vegetation pattern, supposed pollen source locations were obtained for each hour. By comparing supposed hourly pollen origins with the intradiurnal patterns of pollen concentrations, it can be seen that this simple model explains variations in mean pollen concentrations throughout the day. Received: 6 May 1998 / Revised: 24 February 1999 / Accepted: 12 March 1999     

Feeding friend and foe: ample pollen mitigates the effects of pollen theft for a gynodioecious plant,Polemonium foliosissimum (Polemoniaceae)

Background and AimsMost angiosperms rely on pollinators to transport pollen and effect fertilization. While some floral visitors are effective pollinators, others act as thieves, consuming pollen but effecting little pollination in return. The importance of pollen theft in male and female reproductive success has received little attention. Here, we examined if pollen consumption by flies altered pollen receipt and exacerbated pollen limitation for a bumblebee-pollinated plant, Polemonium foliosissimum (Polemoniaceae).MethodsTo examine the effect of pollen-thieving flies, we took a three-pronged approach. First, we used single-visit observations to quantify pollen removal and pollen deposition by flies and bumblebees. Second, we manipulated pollen in the neighbourhood around focal plants in two years to test whether pollen reduction reduced pollen receipt. Third, we combined pollen reduction with hand-pollination to test whether pollen thieving exacerbated pollen limitation. Polemonium foliosissimum is gynodioecious in most populations in the Elk Mountains of central Colorado, USA. Thus, we also tested whether pollen theft affected hermaphrodites and females differently.ResultsFlies removed significantly more pollen and deposited less pollen per visit than did bumblebees. Reduction of pollen in the neighbourhood around focal plants reduced pollen receipt in both years but only nearly significantly so in 2015. In 2016, plants were significantly pollen-limited; hand-pollination significantly increased seeds per fruit for both hermaphrodites and females. However, the reduction of pollen around focal plants did not exacerbate pollen limitation for either hermaphrodites or females.ConclusionsOur results suggest that plants tolerate significant consumption of pollen by thieves and pollinators by producing ample pollen to feed both and fertilize available ovules. Our results demonstrate that pollen limitation in P. foliosissimum is driven by lack of effective pollinators rather than lack of pollen. Teasing out these effects elucidates the relative importance of drivers of reproductive success and thus the expected response to selection by different floral visitors.     

Transport of airborne pollen into the city of Thessaloniki: the effects of wind direction, speed and persistence

We examined the effect of the wind vector analyzed into its three components (direction, speed and persistence), on the circulation of pollen from different plant taxa prominent in the Thessaloniki area for a 4-year period (1996–1999). These plant taxa were Ambrosia spp., Artemisia spp., Chenopodiaceae, Corylus spp., Cupressaceae, Olea europaea, Pinaceae, Platanus spp., Poaceae, Populus spp., Quercus spp., and Urticaceae. Airborne pollen of Cupressaceae, Urticaceae, Quercus spp. and O. europaea make up approximately 70% of the total average annual pollen counts. The set of data that we worked with represented days without precipitation and time intervals during which winds blew from the same direction for at least 4 consecutive hours. We did this in order to study the effect of the different wind components independently of precipitation, and to avoid secondary effects produced by pollen resuspension phenomena. Factorial regression analysis among the summed bi-hourly pollen counts for each taxon and the values of wind speed and persistence per wind direction gave significant results in 22 cases (combinations of plant taxa and wind directions). The pollen concentrations of all taxa correlated significantly with at least one of the three wind components. In seven out of the 22 taxon-wind direction combinations, the pollen counts correlated positively with wind persistence, whereas this was the case for only two of the taxon-wind speed combinations. In seven cases, pollen counts correlated with the interaction effect of wind speed and persistence. This shows the importance of wind persistence in pollen transport, particularly when weak winds prevail for a considerable part of the year, as is the case for Thessaloniki. Medium/long-distance pollen transport was evidenced for Olea (NW, SW directions), Corylus (NW, SW), Poaceae (SW) and Populus (NW).     

Construction,validation, and application of nocturnal pollen transport networks in an agro‐ecosystem: a comparison using light microscopy and DNA metabarcoding

1. Moths are globally relevant as pollinators but nocturnal pollination remains poorly understood. Plant–pollinator interaction networks are traditionally constructed using either flower‐visitor observations or pollen‐transport detection using microscopy. Recent studies have shown the potential of DNA metabarcoding for detecting and identifying pollen‐transport interactions. However, no study has directly compared the realised observations of pollen‐transport networks between DNA metabarcoding and conventional light microscopy. 2. Using matched samples of nocturnal moths, we constructed pollen‐transport networks using two methods: light microscopy and DNA metabarcoding. Focussing on the feeding mouthparts of moths, we developed and provide reproducible methods for merging DNA metabarcoding and ecological network analysis to better understand species interactions. 3. DNA metabarcoding detected pollen on more individual moths, and detected multiple pollen types on more individuals than microscopy, although the average number of pollen types per individual was unchanged. However, after aggregating individuals of each species, metabarcoding detected more interactions per moth species. Pollen‐transport network metrics differed between methods because of variation in the ability of each to detect multiple pollen types per moth and to separate morphologically similar or related pollen. We detected unexpected but plausible moth–plant interactions with metabarcoding, revealing new detail about nocturnal pollination systems. 4. The nocturnal pollination networks observed using metabarcoding and microscopy were similar yet distinct, with implications for network ecologists. Comparisons between networks constructed using metabarcoding and traditional methods should therefore be treated with caution. Nevertheless, the potential applications of metabarcoding for studying plant–pollinator interaction networks are encouraging, especially when investigating understudied pollinators such as moths.     

Temporal and geographical variations in grass pollen seasons in areas of west0ern Europe: an analysis of season dates at sites of the European pollen information system

Geographical and temporal variations in the start dates of grass pollen seasons are described for selected sites of the European Pollen Information Service. Daily average grass pollen counts are derived from Network sites in Finland, the Netherlands, Denmark, United Kingdom, Austria, Italy and Spain, giving a broad longitudinal transect over Western Europe. The study is part of a larger project that also examines annual and regional variations in the severity, timing of the peak and duration of the grass pollen seasons. For several sites, data are available for over twenty years enabling long term trends to be discerned. The analyses show notable contrasts in the progression of the seasons annually with differing lag times occurring between southern and northern sites in various years depending on the weather conditions. The patterns identified provide some insight into geographical differences and temporal trends in the incidence of pollinosis. The paper discusses the main difficulties involved in this type of analysis and notes possibilities for using data from the European Pollen Information service to construct pan European predictive models for pollen seasons. This revised version was published online in June 2006 with corrections to the Cover Date.     

Air pollution exacerbates effect of allergenic pollen proteins of Cassia siamea: a preliminary report

Pollinosis caused by several allergenic proteins in pollen grains has been a major problem of healthcare in most parts of the world. Environmental factors such as air pollution are known to alter the release of allergenic pollen proteins from a variety of the plant species. Cassia siamea is commonly planted along roadsides and in industrialised areas in many parts of India. The present study reports the findings of animal experiments demonstrating the effect of air pollution on the allergenicity of Cassia siamea pollen proteins. Total white blood cell (WBC) count and lymphocyte count were significantly higher in animals that received protein extract of pollen collected from a polluted site compared to those that received protein extract of pollen collected from a non-polluted site. This was concomitant with increased production of IgE antibodies; followed by marked degranulation of mast cell leading to heighten type I hypersensitivity in these animals. These results are important for the development of a consensus linking ever-increasing pollution due to industrialisation and an increase in associated pollinosis.     

Six years of observation of airborne and deposited pollen in central European Russia: first results

The aim of the study was to compare the pollen spectra recorded in rural and urban environments in order to examine whether pollen data obtained in the urban environment could also represent the situation in surrounding rural areas and how urban planting affects the regional pollen spectrum. Daily airborne pollen observations were performed with a volumetric Hirst type trap in Moscow (Russia). Modified Tauber traps were located in the Moscow region. The Tauber traps were installed and treated according to Pollen Monitoring Programme (PMP) guidelines. Spearman correlation coefficients were determined between the percentage of total pollen recorded in the Tauber and Hirst type samplers. The data showed that the Tauber spectra are characterised by higher taxonomic variability, mainly due to the greater occurrence of local herbaceous plants. Pollen types registered by Hirst type samplers were predominantly represented by anemophilous arboreal species, used in urban planting. A significant correlation for annual sums was only observed for Betula pollen, the most abundant component in both sampler types. Data obtained in big cities for dominant taxa can be extrapolated to the surrounding rural territories. The simultaneous use of volumetric and gravimetric samplers can be very useful for palaeo-ecological studies to evaluate the presence of local and transported pollen.     

Intrusion of airborne pollen through open windows and doors

The importance of the transport of pollen by air movement into houses was evaluated using six to eight simultaneously collecting rotorod-type samplers, creating either a sampler line from outdoors to inside the room, or a sampler grid inside a room. The number of incoming pollen grains was highly dependent on the outdoor concentration. The highest concentrations inside (1–2 m distance) and outside (1 m) the room were 600 and 3,250 grains/m³, respectively, in the Betula pollen season and 1,980 and 5,080 grains/m³ in the Pinus season. The pollen concentration and the indoor/outdoor (I/O) ratio decreased as the distance from the ventilation opening increased. Inside the room at a distance of 1–2 m 28%, and at a distance of 3–5 m 12%, of the outside concentration was recorded. In the lower part of the opening the mean proportion was 63% and in the upper part of the opening it was 40%. Efficient ventilation with two open windows increased the I/O ratio and enabled the pollen to spread throughout the room. During the Pinus pollen season 3–35% of the outdoor concentration was simultaneously recorded at six locations inside the room with two open windows and only 0.1–3.6% with one open window. At the same point in the room the I/O ratio varied from <1 to 35%, depending on the sampling conditions. Only a minor effect on the I/O ratio was found between small and large ventilation windows and the door, although it was expected that more air and pollen grains would come indoors through a larger opening.