Free orbicules of Cupressaceae detected in daily aerobiological samples by optical and confocal microscopy

Members of Cupressaceae and Taxaceae are known to release large amounts of highly allergenic pollen grains into the atmosphere, which are responsible for the onset of pollinosis in many countries throughout the world. In addition to pollen grains, their pollen sacs produce orbicules, which are submicron particles reported to carry allergens and which are potentially able to reach much further down the respiratory tract than pollen grains. Previous research has postulated the presence of orbicules in the atmosphere; however, direct observations have not yet been reported. The aim of this research was to provide the first direct evidence that Cupressaceae orbicules are released into the atmosphere by detecting them in daily aerobiological samples. We observed pollen sacs, pollen grains, and orbicules of nine species of Cupressaceae using scanning electron microscope (SEM). We then used a light and confocal microscope, to examine daily aerobiological samples. Under SEM, we measured the orbicule size (0.494–0.777 µm) and detected unknown nanometric particles (130–200 nm). Under the light microscope, aerobiological samples showed clusters of stained dots surrounding the pollen grains of Cupressaceae. Under the confocal microscope, the same clusters were resolved into submicron particles with the same autofluorescence as the pollen grains. These features enabled us to identify them as orbicules. We believe that our findings help to explain the onset of pollinosis and allergic asthma related to Cupressaceae pollen grains in many countries, even before pollen grains are actually detected or after they are no longer observed in aerobiological monitoring samples.     

Airborne pollen patterns in Mar del Plata atmosphere (Argentina) and its relationship with meteorological conditions

This work is part of a series of aerobiological researches conducted in the city of Mar del Plata, Argentina. The annual, seasonal and daily features of the pollen cloud were analyzed over 2 years, together with the effects of the meteorological variables. Cupressaceae amounted to 75 and 54% of the annual total in each year, and it was analyzed separately due to its proven overrepresentation. Eighty-five pollen types were registered during the 2 years, bearing a similar annual pattern (more than 50% from August to November, December also being an important month). Arboreal pollen predominated between late winter and spring (61 and 49% of the annual total), while non-arboreal pollen did so from late spring to summer. The observed seasonality was significantly correlated with the monthly mean temperatures and associated with its effect on phenology. No significant differences along the day were detected between years and 50% of the daily total was recorded between 1000 and 1600 hours. The maximum concentration was registered at 1000 hours in spring and summer, but later in autumn and winter. Regarding the hourly scale, the significant correlation of total pollen with temperature and wind speed was positive, albeit negative with relative humidity. These effects are linked to airborne pollen release and transport. The role rainfall plays on airborne pollen is discussed. To interpret the results, it is important to consider the time scale at which the aerobiological phenomenon is analyzed.     

A principal component regression model to forecast airborne concentration of Cupressaceae pollen in the city of Granada (SE Spain), during 1995–2006

The problem of developing a 2-week-on ahead forecast of atmospheric cypress pollen levels is tackled in this paper by developing a principal component multiple regression model involving several climatic variables. The efficacy of the proposed model is validated by means of an application to real data of Cupressaceae pollen concentration in the city of Granada (southeast of Spain). The model was applied to data from 11 consecutive years (1995–2005), with 2006 being used to validate the forecasts. Based on the work of different authors, factors as temperature, humidity, hours of sun and wind speed were incorporated in the model. This methodology explains approximately 75–80% of the variability in the airborne Cupressaceae pollen concentration.     

An aerobiological study of Urticaceae pollen in the city of Granada (S. Spain): Correlation with meteorological parameters

An aerobiological study was made of Urticaceae pollen in the city of Granada, relating the mean values of the daily counts to meteorological parameters. Sampling was carried out with a Burkard seven‐day‐recording spore trap from October 1992 to September 1997. This pollen type has an extremely long main pollen season (MPS), with maximum counts in (January) February, March and April, causing numerous cases of human pollinosis throughout the entire Mediterranean region, including Granada. A highly constant intradiurnal variation pattern was obtained showing that the maximum peaks usually occur between 12.00 and 20.00. According to Spearman's correlation coefficient, during the pre‐peak period the parameters which have the greatest effect on the levels of this type of pollen are daily and accumulated temperature and sunshine, accumulated rainfall, and wind direction from the third quadrant; during the post‐peak period these same variables presented significantly negative coefficients. Daily rainfall and relative humidity presented negative coefficients during the entire MPS. The maximum daily temperature was the variable which provided the closest match with the theoretical predictive pattern presented here.     

<Emphasis Type="Italic">Cyperaceae</Emphasis> and <Emphasis Type="Italic">Juncaceae</Emphasis> pollination measured in the air at two sites in SW Spain

The presence of airborne Cyperaceae and Juncaceae pollen was quantified using volumetric aerobiological traps over a 10-year period at two sites in SW Spain separated by 60 km (Badajoz 10 years, Mérida 3 years). The Pearson correlation coefficient was calculated between the daily and hourly concentrations. The values of the principal meteorological parameters—temperature, rainfall, relative humidity, and speed and direction of the wind—were calculated during the study period, and with the accumulated values corresponding to the period prior to pollination. The beginning and ending dates and the duration of the main pollen season (MPS) were also analyzed. Even though both families are anemophilous, the presence of their pollens in the air was low. The annual accumulated daily concentrations were in the range 1.8–15.8 for Juncaceae and 111.8–473.9 for Cyperaceae—values far lower than any other anemophilous pollen type. The Cyperaceae pollen concentration peaked between 09:00 and 12:00. The meteorological factor most closely related to its daily variations was found to be the wind direction, showing that location of the sources is of great importance. The results lend support to the hypothesis of a limitation of allogamous reproduction in favour of vegetative multiplication in both families. Nevertheless, the principal source of the airborne Cyperaceae pollen was found to be Scirpus holoschoenus, whose pollen is distinguishable from the rest. Hence, because of its large production of pollen, this species can be characterized as anemophilous and allogamous. Rainfall in the preceding autumn seemed to be responsible for the amount of Cyperaceae pollen in the air, since a lack of rain was found to be associated with lower densities in the traps. For the Juncaceae, it seems that the temperatures of the preceding December constituted the most limiting meteorological parameter.     

Comparative study of the effect of distance on the daily and hourly pollen counts in a city in the south-western Iberian Peninsula

The airborne pollen concentration in the city of Badajoz was measured in two locations 2.9 km apart. The measurements were taken from January to June between 2009 and 2012 using Hirst-type volumetric aerobiological samplers. One sampler was placed at the Faculty of Science (FS) and the other at the Agricultural Engineering School (AES) of the University of Extremadura, Spain, on terraces located 16 and 6 m above ground, respectively. The two sets of hourly and daily pollen concentrations were compared regarding the meteorological parameters and the distribution of local pollen sources. A total of 46 pollen types were counted, with a mean total concentration of 43 pollen grains/m³ in the winter and 336 pollen grains/m³ in the spring. In the winter, pollen grains from (in decreasing order) Cupressaceae, Fraxinus–Phillyrea, Urticaceae spp., Alnus glutinosa and Urtica membranacea types represented 77 % of the total. In the spring, 89 % of the total was represented by pollen grains from (in decreasing order) Quercus, Poaceae, Olea, Pinaceae and Plantago. The FS/AES ratio was 0.57 in the winter and 1.31 in the spring. While a Wilcoxon test applied to the daily total concentration data showed statistically significant differences between the two sites, a correlation study based on the Spearman coefficient showed statistically significant correlations in both the winter and spring. The results were similar when comparing the separate pollen types, except for Urticaceae spp., which showed no statistically significant correlation. The meteorological data studied showed a statistically significant correlation with the daily concentrations. A comparison of the hourly concentration data showed no correspondence with the time of maximum concentration. The local distribution of pollen sources explained some of the differences found between the two sites. Overall, the results indicate that a single aerobiological sampler may be sufficiently representative to register the daily pollen grain data of an urban area of approximately 3 km or greater in diameter, although it underestimates the influence of heterogeneity in the distribution of the local flora.     

The reliability of geostatistic interpolation in olive field floral phenology

Floral phenology, as most of natural phenomena, shows, as an inherent feature, a high degree of spatial continuity. Geostatistics are a family of statistics that describe correlations through space/time and they can be used for both quantifying spatial correlation and interpolating no monitored sampling points. The combined use of Geographical Information Systems (GIS) and geostatistics can be an essential tool for spatial analysis in phenological and aerobiological studies. In the present work, Kriging interpolation by using linear geostatistic analysis has allow us to estimate phenological data of a wide olive crop area of the province of Cordoba (Andalusia, Southern Spain), on the basis of a limited number of␣phenology sampling points. The main a priori hypothesis was that 7 traditionally observed sampling points (with 10 olive trees in each site) uniformly distributed through the main olive crop areas could be enough for interpolating phenological information of the whole Cordoba olive area. Geostatistical results reject this hypothesis. The optimum/minimum number and location of sampling points was determined in 13 sites (including the original 7 sites and 6 new sites). The obtained phenological maps will improve␣olive pollen aerobiological information and forecasting in the area. The application of such new combined space analysis tools on floral phenology allows optimising human and economic resources on field phenology campaigns. Moreover, an appropriate use of GIS and geostatistic software to create phenological maps will be an essential complement in pollen aerobiological studies, given the increased interest in obtaining automatic aerobiological forecasting maps.     

Study on allergenicity of Thuja orientalis pollen grains in rat

Cupressaceae pollen allergy is an important cause of pollen allergy throughout the world. Prevalence of allergy to Cupressaceae pollen has increased significantly during the winter over the past 3 decades because of extensive planting of cypress trees for different purposes. Thuja orientalis (Cupressaceae) is a naturally grown plant in Iran and is widely cultivated as a common ornamental plant in this country and other ones. Allergenicity of its pollen has been established, but to this day no allergenic component has been detected. The aim of this research is to study allergenicity and evaluate the immunoglobulin E reactivity to T. orientalis pollen extracts. Pollen grains were directly collected from mature male cones of trees. Pollen proteins were extracted and were analyzed by SDS-PAGE. Total protein content of pollen extracts was measured by Bradford assay. Immunoblotting using the serum of sensitized rats showed a single immunogenic band at about 44KD in pollen extracts. Result of this research proved that pollen grains of T. orientalis are allergenic.     

In Memoriam Siwert Nilsson 1933–2002

Since the beginning of aeropalynology, aerobiologists have tried to develop models able to predict the pollen load of the atmosphere to help physicians and patients suffering pollen allergies to improve the efficiency of pollen allergies prophylaxis. Some models have been developed by aerobiological services and are used for real time predictions, now provided online. They are statistical models and require the pollen load of the previous days as input, as well as statistics on the pollen load of the previous years, daily meteorological data and geographic information on the pollen sources. Here we propose to use process-based phenological models to predict the time course of the pollen load of the atmosphere using solely daily mean temperatures as input. The model proposed has been fitted and validated for 13 major allergenic taxa (Alnus, Artemisia, Betula, Castanea, Corylus, Cupressaceae, Olea, Plantago, Platanus, Populus, Poaceae, Quercus and Tilia) in different areas of France and Spain. The model has been integrated into a freeware called PPF (Positive Pollen Forecast), which will be made available from the 5^th Framework EU project POSITIVE web page at: http://www.forst.tu-muenchen.de/LST/METEOR/positive/.     

Gramineae hay fever,and aerobiological and clinical investigation in Palermo; Sicily

Summary Epidemiological and aerobiological observations (1987;1989) have been carried out for three years in order to search the existing relationship between the Gramineae's daily pollen concentration in Palermo's atmosphere and the number of hay fever cases due to such pollen.The aerobiological data were obtained with a 2000 VPPS volumetric sampler. Clinical research was performed on 555 hay fever patients treated in our ambulatory over a three-year period (1987–1989).These data, elaborated by a seven day running mean method and correlated with epidemiological data, evidenced that three of the pollen families in our territory are clinically important: Urticaceae,Parietaria prevailing among them, Gramineae and Oleaceae,Olea europaea prevailing among them.The Gramineae are the second most important allergenic pollen (32,08% of all the pollinosis) whereas, as far as its concentration in the atmosphere is concerned; it ranks third followingParietaria and Oleaceae.     

Use of <Emphasis Type="Italic">Quercus ilex</Emphasis> subsp. <Emphasis Type="Italic">ballota</Emphasis> phenological and pollen-production data for interpreting <Emphasis Type="Italic">Quercus</Emphasis> pollen curves

Although aerobiological data are often used in phenological research as an indicator of flowering, airborne pollen concentrations are influenced by a number of factors that could affect pollen curves. This paper reports on a study of various aspects of reproductive biology in Q. ilex subsp. ballota, together with environmental factors influencing pollen release and transport, with a view of achieving reliable interpretation of Quercus pollen curves in Ourense (NW Spain). Aerobiological data were recorded from 2002 to 2004 at two sites in the province of Ourense. From 1st February to the end of the flowering period, phenological observations were carried out on 19 trees from the Q. ilex subsp. ballota population found in the Ourense area. Pollen production was calculated for the same trees. The chilling and heating requirements for triggering development were also calculated. The mean flowering period lasted 11-15 days. Reduced pollen output per catkin and, especially, a reduced number of catkins per tree in 2003 and 2004, prompted a marked decline in overall pollen production. Major differences observed in Q. ilex subsp. ballota pollen curves were attributed to the considerable influence both of weather conditions during pollination and pollen production. In years with high pollen production and weather conditions favouring pollen release, Q. ilex subsp. ballota contributed almost 10% to the total Quercus pollen curve. Around 20% of the pollen trapped was captured before or after flowering periods.     

An integrated use of aerobiological and phenological data to analyse flowering in olive groves

Aerobiological and phenological investigation has been linked to analyse the flowering phenomena of olive (Olea europaea L.), a tree of economic importance, in Umbria, central Italy.

Olive tree flowering was analysed by phenological observations in olive groves. Aerobiological monitoring was carried out with pollen traps that captured the pollen grains in the atmosphere by remote distances. This combined study of flowering was useful in overcoming the principal limit of phenological observations caused by their geographical narrowness.

Eleven phenological stations in Perugia province were used and periodic observations were carried out during the flowering season. Two aerobiological stations located near the cities of Perugia and Spoleto were utilized to trap olive pollen grains in the atmosphere.

Graphic and statistical analyses were used in order to study flowering dynamics, and to compare the methodological approaches.

The phenological areas, according to their characteristics, were divided in two clusters each one related to a different pollen monitoring station. Statistical analyses showed the presence of a three ‐ four‐day period during which pollen released in the groves is persistent in the narrowness and resulted in a high correlation with the pollen monitored by the pollen traps. On the other hand, the phenological data considered along with the daily pollen concentrations permit the pollen peaks to be attributed to particular olive areas or olive cultivars. This demonstrates that both methodologies can be used effectively as integrated research tools in this kind of study.     

Correlation between aerobiological and phytogeographical investigations in the Florence area

Summary The aim of our research work was to evaluate the results from our pollen count (p.c.) carried out at the Allergological Centre of Nuovo Osp. San Giovanni di Dio in Florence, in the light of orogeographical, phytogeographical and meteorological data. The highest p.c. was yielded by Graminaceae which appear in small quantities early in the year but increase considereably starting from April. Oleaceae too showed high pollen concentration but over a shorter lenght of time. The p.c. for Compositae and Urticaceae was lower. It should be noted that Cupressaceae yielded constantly high p.c. in February–March. Investigations on the allergenic flora of the surroundings were carried out concentrically in order to find out the closest and the farthest sources of pollen production. The species investigated come under 7 types of vegetation: xerophilous woods, mesophilous woods, conifers woods, parks and gardens, rural areas, meadows, border areas-weeds. A phytosociological study of the types of vegetation has made it possible to evaluate the single species quantitatively. The different sources of pollen production can be spotted by mapping these types of vegetation. The seasonal characterization of pollen production is better defined by means of periodical phenological checking of the existing allergenic flora along carefully planned routes through the areas next to the sampling station.      

Phenological records as a complement to aerobiological data

Phenological studies in combination with aerobiological studies enable one to observe the relationship between the release of pollen and its presence in the atmosphere. To obtain a suitable comparison between the daily variation of airborne pollen concentrations and flowering, it is necessary for the level of accuracy of both sets of data to be as similar as possible. To analyse the correlation between locally observed flowering data and pollen counts in pollen traps in order to set pollen information forecasts, pollen was sampled using a Burkard volumetric pollen trap working continuously from May 1993. For the phenological study we selected the main pollen sources of the six pollen types most abundant in our area: Cupressaceae, Platanus, Quercus, Plantago, Olea, and Poaceae with a total of 35 species. We selected seven sites to register flowering or pollination, two with semi-natural vegetation, the rest being urban sites. The sites were visited weekly from March to June in 2007, and from January to June in 2008 and 2009. Pollen shedding was checked at each visit, and recorded as the percentage of flowers or microsporangia in that state. There was an association between flowering phenology and airborne pollen records for some of the pollen types (Platanus, Quercus, Olea and Plantago). Nevertheless, for the other types (Cupressaceae and Poaceae) the flowering and airborne pollen peaks did not coincide, with up to 1 week difference in phase. Some arguments are put forward in explanation of this phenomenon. Phenological studies have shown that airborne pollen results from both local and distant sources, although the pollen peaks usually appear when local sources are shedding the greatest amounts of pollen. Resuspension phenomena are probably more important than long-distance transport in explaining the presence of airborne pollen outside the flowering period. This information could be used to improve pollen forecasts.     

First aerobiological study in Mediterranean part of Croatia (Dalmatia): pollen spectrum and seasonal dynamics in the air of Split

The aim of this study was to investigate aerobiological dynamics of pollen in the Mediterranean part of Croatia in the air of the city of Split. Pollen monitoring during the period from 2005 to 2013 was performed using a Hirst volumetric trap. Among the identified pollen of 50 taxa, 21 were allergenic. The average annual pollen index was 33,513. Three pollination seasons were established: early winter season dominated by tree pollen, spring–summer season dominated by herbaceous plants and summer–autumn season with lower amounts of Parietaria and Cupressaceae pollen. According to the abundance, the main taxa were: Cupressaceae, Parietaria/Urtica, Pinus, Quercus, Olea, Carpinus/Ostrya, Poaceae, Platanus and Ambrosia. The annual pollen index together with the daily maximum concentrations showed an upward trend for selected taxa during the study period. The highest monthly pollen index and the highest biodiversity were recorded in April and the lowest during the late autumn and winter months. The pollen calendar created for the city of Split confirmed Mediterranean features of the pollen spectrum. The longest pollen seasons were recorded for Cupressaceae, Parietaria/Urtica and Poaceae pollen types. The correlations between pollen concentrations and meteorological parameters were analyzed. The correlations between pollen concentrations and temperature were positive, while the humidity and the precipitation mostly showed negative influence.     

Aerobiology of Fagaceae pollen in Trieste (NE Italy)

The aerobiological behaviour of Fagaceae in Trieste and the correlations with the meteorological parameters were examined. Airborne pollen grains of Castanea, Fagus and Quercus were collected from 1990 to 2003 using a Hirst type spore trap. The main pollen season (MPS) takes place in April and May for Quercus and Fagus, in June and July for Castanea. The highest values occur in year 1993 for Quercus, in 1998 for Castanea and in 1992 for Fagus. The Fagaceae content of the air is mainly due to Quercus and Castanea pollen, Fagus usually having a scarce pollen shedding in Trieste. The highest counts of Fagaceae pollen grains are found from late April to mid May and are mainly due to the pollen shedding of oaks. The cumulative counts vary over the years, with a mean value of 2.719 pollen grains, a lowest total of 1.341 in 2002 and a highest total of 4.704 in 1993. No positive nor negative long-term trends in pollen shedding are found. No cyclic variations were observed. Spearman’s correlation was used to establish the relationship between the daily pollen counts and the daily meteorological data. Daily pollen concentrations present sometimes positive correlation with temperature, negative with rainfall and wind speed, and no correlation with humidity. Fagus and Quercus start dates result positively correlated between themselves. Significant correlations are found between the start of MPS and the mean and maximum temperature in March for Fagus and Quercus, and May for Castanea.     

Pollen from tree and shrub taxa in the atmosphere of Caxias do Sul (Rio Grande do Sul, Brazil)

An aerobiological study has been carried out in the region of Caxias do Sul in southern Brazil. Pollen monitoring was performed from January 1, 2001 through to December 31, 2002. A total of 30,469 pollen grains were collected during this period, and 40 pollen types were identified; of these, 23,389 pollen grains, representing 29 pollen types, originated from tree and shrub taxa. The maximum pollen concentration was registered in August 2001 and October 2002. In the study area, the pollen type Mimosa scabrella (18.8%) was much more abundant than all of the pollen types from tree and shrub taxa, such as: Urticaceae (18.4%), Myrtaceae (10.2%), Cupressaceae (7.7%), Myrsine (4.8%), Sorocea (3.9%), Pinaceae (2.9%), Asteraceae (2.2%) and Ricinus (2.1%). These nine pollen types accounted for the largest pollen concentrations of all the tree and shrub taxa. The pollen types Carya, Melastomataceae, Mimosa scabrella, Myrsine and Sorocea are reported for the first time in an aerobiological study in Brazil.     

Forecasting model of <Emphasis Type="Italic">Corylus</Emphasis>, <Emphasis Type="Italic">Alnus</Emphasis>, and <Emphasis Type="Italic">Betula</Emphasis> pollen concentration levels using spatiotemporal correlation properties of pollen count

The aim of the study was to create and evaluate models for predicting high levels of daily pollen concentration of Corylus, Alnus, and Betula using a spatiotemporal correlation of pollen count. For each taxon, a high pollen count level was established according to the first allergy symptoms during exposure. The dataset was divided into a training set and a test set, using a stratified random split. For each taxon and city, the model was built using a random forest method. Corylus models performed poorly. However, the study revealed the possibility of predicting with substantial accuracy the occurrence of days with high pollen concentrations of Alnus and Betula using past pollen count data from monitoring sites. These results can be used for building (1) simpler models, which require data only from aerobiological monitoring sites, and (2) combined meteorological and aerobiological models for predicting high levels of pollen concentration.     

Phenological and aerobiological monitoring of allergenic flora in padua (Italy). Preliminary data

A phenological study on allergenic plants was carried out in Padua during 1995 in order to identify spontaneous and cultivated allergenic species in an urban area and their distribution, and to evaluate the relationship between anthesis length and airborne pollen concentrations. In some cases, there was no temporal overlap between phenological and aerobiological data, in particular forCorylaceae, Betulaceae, Chenopodiaceae, Amaranthaceae, Polygonaceae and Fagaceae.     

Evidences of olive pollination date variations in relation to spring temperature trends

Airborne pollen concentration patterns reflect flowering phenology of a given species, and it may be a sensitive regional indicator in climate change studies. This paper presents the relationship between a strategic biological event, such as olive flowering, and the air temperature trend, registered over a large scale (1982–2007) in the Umbria region. The aim of the study was to determine relationships between phenological behaviour (flowering) of olive trees and the air temperature trend (1982–2007) in the Umbria region. The phenological data on flowering phase were registered indirectly through an aerobiological monitoring technique. The obtained results showed a strong relationship between phenology and thermal trend. This characteristic was confirmed from results of correlations between temperature (mean temperature from 1st March) and flowering dates, especially that of full flowering (r = −0.9297). Moreover, the results showed an advance trend of 6, 8 and 10 days, respectively of start, full and end of flowering dates. The advance of the recorded flowering time in this period is to ascribe mainly to the increase of mean temperature and above all to that registered in months of May and June.