The Macroecology of Airborne Pollen in Australian and New Zealand Urban Areas

he composition and relative abundance of airborne pollen in urban areas of Australia and New Zealand are strongly influenced by geographical location, climate and land use. There is mounting evidence that the diversity and quality of airborne pollen is substantially modified by climate change and land-use yet there are insufficient data to project the future nature of these changes. Our study highlights the need for long-term aerobiological monitoring in Australian and New Zealand urban areas in a systematic, standardised, and sustained way, and provides a framework for targeting the most clinically significant taxa in terms of abundance, allergenic effects and public health burden.

Total transcriptome, proteome, and allergome of Johnson grass pollen, which is important for allergic rhinitis in subtropical regions

Background Genomic data are lacking for many allergen sources. To circumvent this limitation, we implemented a strategy to reveal the repertoire of pollen allergens of a grass with clinical importance in subtropical regions, where an increasing proportion of the world's population resides. Objective We sought to identify and immunologically characterize the allergenic components of the Panicoideae Johnson grass pollen (JGP; Sorghum halepense). Methods The total pollen transcriptome, proteome, and allergome of JGP were documented. Serum IgE reactivities with pollen and purified allergens were assessed in 64 patients with grass pollen allergy from a subtropical region. Results Purified Sor h 1 and Sor h 13 were identified as clinically important allergen components of JGP with serum IgE reactivity in 49 (76%) and 28 (43.8%), respectively, of patients with grass pollen allergy. Within whole JGP, multiple cDNA transcripts and peptide spectra belonging to grass pollen allergen families 1, 2, 4, 7, 11, 12, 13, and 25 were identified. Pollen allergens restricted to subtropical grasses (groups 22-24) were also present within the JGP transcriptome and proteome. Mass spectrometry confirmed the IgE-reactive components of JGP included isoforms of Sor h 1, Sor h 2, Sor h 13, and Sor h 23. Conclusion Our integrated molecular approach revealed qualitative differences between the allergenic components of JGP and temperate grass pollens. Knowledge of these newly identified allergens has the potential to improve specific diagnosis and allergen immunotherapy treatment for patients with grass pollen allergy in subtropical regions and reduce the burden of allergic respiratory disease globally.

An immunodiagnostic assay for quantitation of specific ige to the major pollen allergen component, pas n 1, of the subtropical bahia grass

Background Pollens of the Panicoideae subfamily of grasses including Bahia (Paspalum notatum) are important allergen sources in subtropical regions of the world. An assay for specific IgE to the major molecular allergenic component, Pas n 1, of Bahia grass pollen (BaGP) would have immunodiagnostic utility for patients with pollen allergy in these regions. Methods Biotinylated Pas n 1 purified from BaGP was coated onto streptavidin ImmunoCAPs. Subjects were assessed by clinical history of allergic rhinitis and skin prick test (SPT) to aeroallergens. Serum total, BaGP-specific and Pas n 1-specific IgE were measured. Results: Pas n 1 IgE concentrations were highly correlated with BaGP SPT (r = 0.795, p < 0.0001) and BaGP IgE (r = 0.915, p < 0.0001). At 0.23 kU/l Pas n 1 IgE, the diagnostic sensitivity (92.4%) and specificity (93.1%) for the detection of BaGP allergy was high (area under receiver operator curve 0.960, p < 0.0001). The median concentrations of Pas n 1 IgE in non-Atopic subjects (0.01 kU/l, n = 67) and those with other allergies (0.02 kU/l, n = 59) showed no inter-group difference, whilst grass pollen-Allergic patients with allergic rhinitis showed elevated Pas n 1 IgE (6.71 kU/l, n = 182, p < 0.0001). The inter-Assay coefficient of variation for the BaGP-Allergic serum pool was 6.92%. Conclusions Pas n 1 IgE appears to account for most of the BaGP-specific IgE. This molecular component immunoassay for Pas n 1 IgE has potential utility to improve the sensitivity and accuracy of diagnosis of BaGP allergy for patients in subtropical regions.

Grass pollen allergens globally: The contribution of subtropical grasses to burden of allergic respiratory diseases

Grass pollens of the temperate (Pooideae) subfamily and subtropical subfamilies of grasses are major aeroallergen sources worldwide. The subtropical Chloridoideae (e.g. Cynodon dactylon; Bermuda grass) and Panicoideae (e.g. Paspalum notatum; Bahia grass) species are abundant in parts of Africa, India, Asia, Australia and the Americas, where a large and increasing proportion of the world's population abide. These grasses are phylogenetically and ecologically distinct from temperate grasses. With the advent of global warming, it is conceivable that the geographic distribution of subtropical grasses and the contribution of their pollen to the burden of allergic rhinitis and asthma will increase. This review aims to provide a comprehensive synthesis of the current global knowledge of (i) regional variation in allergic sensitivity to subtropical grass pollens, (ii) molecular allergenic components of subtropical grass pollens and (iii) allergic responses to subtropical grass pollen allergens in relevant populations. Patients from subtropical regions of the world show higher allergic sensitivity to grass pollens of Chloridoideae and Panicoideae grasses, than to temperate grass pollens. The group 1 allergens are amongst the allergen components of subtropical grass pollens, but the group 5 allergens, by which temperate grass pollen extracts are standardized for allergen content, appear to be absent from both subfamilies of subtropical grasses. Whilst there are shared allergenic components and antigenic determinants, there are additional clinically relevant subfamily-specific differences, at T- and B-cell levels, between pollen allergens of subtropical and temperate grasses. Differential immune recognition of subtropical grass pollens is likely to impact upon the efficacy of allergen immunotherapy of patients who are primarily sensitized to subtropical grass pollens. The literature reviewed herein highlights the clinical need to standardize allergen preparations for both types of subtropical grass pollens to achieve optimal diagnosis and treatment of patients with allergic respiratory disease in subtropical regions of the world. © 2014 John Wiley & Sons Ltd.

The dominant 55kDa allergen of the subtropical Bahia grass (Paspalum notatum) pollen is a group 13 pollen allergen, Pas n 13

Bahia grass, Paspalum notatum, is an important pollen allergen source with a long season of pollination and wide distribution in subtropical and temperate regions. We aimed to characterize the 55. kDa allergen of Bahia grass pollen (BaGP) and ascertain its clinical importance. BaGP extract was separated by 2D-PAGE and immunoblotted with serum IgE of a grass pollen-allergic patient. The amino-terminal protein sequence of the predominant allergen isoform at 55. kDa had similarity with the group 13 allergens of Timothy grass and maize pollen, Phl p 13 and Zea m 13. Four sequences obtained by rapid amplification of the allergen cDNA ends represented multiple isoforms of Pas n 13. The predicted full length cDNA for Pas n 13 encoded a 423 amino acid glycoprotein including a signal peptide of 28 residues and with a predicted pI of 7.0. Tandem mass spectrometry of tryptic peptides of 2D gel spots identified peptides specific to the deduced amino acid sequence for each of the four Pas n 13 cDNA, representing 47% of the predicted mature protein sequence of Pas n 13. There was 80.6% and 72.6% amino acid identity with Zea m 13 and Phl p 13, respectively. Reactivity with a Phl p 13-specific monoclonal antibody AF6 supported designation of this allergen as Pas n 13. The allergen was purified from BaGP extract by ammonium sulphate precipitation, hydrophobic interaction and size exclusion chromatography. Purified Pas n 13 reacted with serum IgE of 34 of 71 (48%) grass pollen-allergic patients and specifically inhibited IgE reactivity with the 55. kDa band of BaGP for two grass pollen-allergic donors. Four isoforms of Pas n 13 from pI 6.3-7.8 had IgE-reactivity with grass pollen allergic sera. The allergenic activity of purified Pas n 13 was demonstrated by activation of basophils from whole blood of three grass pollen-allergic donors tested but not control donors. Pas n 13 is thus a clinically relevant pollen allergen of the subtropical Bahia grass likely to be important in eliciting seasonal allergic rhinitis and asthma in grass pollen-allergic patients.

IgE cross-reactivity between Pas n 1 of Bahia grass pollen and other group 1 grass pollen allergens

Background The subtropical Bahia grass (Paspalum notatum) is an important source of pollen allergens with an extended season of pollination and wide distribution in warmer climates. The immunological relationship between pollen allergens of Bahia grass and temperate grasses is unresolved. Methods Serum IgE reactivity of grass pollen-allergic patients with Bahia, Ryegrass and Bermuda grass pollen extracts and their purified group 1 allergens, Pas n 1, Lol p 1 and Cyn d 1, were compared by immunoblotting, ELISA, inhibition ELISA, basophil activation by flow cytometry and molecular modeling. Results Differences in antibody recognition of allergenic components between Bahia grass and Ryegrass pollen were observed by immunoblotting. Eight grass pollen-allergic patients from a temperate region showed greater serum IgE reactivity with Ryegrass pollen than Bahia grass by ELISA. For seven of these sera, Ryegrass pollen inhibited IgE reactivity with Bahia grass pollen but not the converse. For 51 sera from grass pollen-allergic patients in this temperate region, IgE reactivity with Lol p 1 was greater than Pas n 1 or Cyn d 1. IgE reactivity with Lol p 1 was not inhibited by Pas n 1 or Cyn d 1, but Pas n 1 IgE reactivity was inhibited by Lol p 1. Two group 1 grass pollen allergen-specific mAb distinguished between temperate and subtropical grass pollens. Basophil activation for three patients tested was greater by Ryegrass pollen than Bahia or Bermuda grass, and by Lol p 1 than Pas n 1 or Cyn d 1. In contrast, two patients from a subtropical region had higher serum IgE reactivity with Bahia grass pollen than Ryegrass and Bahia grass pollen inhibited IgE reactivity with Ryegrass. A structural model of Pas n 1 showed amino acids implicated in IgE epitopes of other group 1 allergens were juxtaposed on the surface. Conclusion Allergens from subtropical Bahia grass pollen, including Pas n 1, share antigenic determinants with temperate grass pollen allergens, but patients exhibit higher serum IgE reactivity to their locally predominant grass pollen. Basophil activation by Bahia grass pollen and Pas n 1 in patients from a temperate climate indicates clinically relevant cross-sensitization between temperate and subtropical grass pollens.

Standardization of pollen allergens of Parthenium hysterophoms and selection of an in-house reference extract

A standardized in-house reference extract from the pollen of Parthenium hysterophorus, which is responsible for the high incidence of allergic rhinitis in India, was generated and examined by skin test, radio-allergosorbent test inhibition and isoelectric focusing. Parthenium reference allergen discs and positive reference serum were also generated. These reference reagents could not only be used for the quantitation of Parthenium-specific IgE in the sera of rhinitis patients but also for the evaluation of allergenic activity (relative potency and lot-to-lot variation) of different batches of Parthenium pollen.

Immunochemical characterization of rapid and slowly released allergens from the pollen of Parthenium hysterophorus

Allergens from the pollen of Parthenium hysterophorus (American feverfew), responsible for high incidence of allergic rhinitis, were found by immunoprint analysis to be localized on the surface of the pollen grains. The allergens were released very rapidly when extracted in vitro. The allergenic activity of the rapid (10 s) and slowly (20 h) released pollen proteins was comparable by in vivo skin test and ELISA inhibition assay. The isoelectric focusing patterns of the rapid and slowly released proteins, were also identical. SDS-PAGE and Western blot analysis revealed that all the major pollen allergens with molecular weight 14, 28, 31, 37 and 45 kDa were eluted within 10 s of extraction. Periodate-Schiff staining showed that the 28, 31 and 45 kDa components of the pollen extract are glycoproteins. The pollen allergens released after different periods of extraction lost 75% of IgE binding activity when subjected to in situ sodium m-periodate oxidation under controlled conditions, while 80% of the allergenic activity was still retained after extensive proteolysis. Our results support the clinical observation of a rapid onset of symptoms of allergic rhinitis in patients sensitive to Parthenium pollen, mediated predominantly by glycoproteins.

The Long Range Transport of Birch (Betula) Pollen from Poland and Germany Causes Significant Pre-season Concentrations in Denmark

Background Birch pollen is highly allergic and has the potential for episodically long range transport. Such episodes will in general occur out of the main pollen season. During that time allergy patients are unprotected and high pollen concentrations will therefore have a full allergenic impact. Objective To show that Denmark obtains significant quantities of birch pollen from Poland or Germany before the local trees start to flower. Methods Simultaneous observations of pollen concentrations and phenology in the potential source area in Poland as well as in Denmark were performed in 2006. The Danish pollen records from 2000-2006 were analysed for possible long range transport episodes and analysed with trajectories in combination with a birch tree source map. Results In 2006 high pollen concentrations were observed in Denmark with bi-hourly concentrations above 500 grains/ m3 before the local trees began to flower. Poland was identified as a source region. The analysis of the historical pollen record from Copenhagen shows significant pre-seasonal pollen episodes almost every year from 2000-2006. In all episodes trajectory analysis identified Germany or Poland as source regions. Conclusion Denmark obtains significant pre-seasonal quantities of birch pollen from either Poland or Germany almost every year. Forecasting of birch pollen quantities relevant to allergy patients must therefore take into account long-range transport. This cannot be based on measured concentrations in Denmark. The most effective way to improve the current Danish pollen forecasts is to extend the current forecasts with atmospheric transport models that take into account pollen emission and transport from countries such as Germany and Poland. Unless long range transport is taken into account pre-seasonal pollen episodes will have a full allergic impact, as the allergy patients in general will be unprotected during that time.

Regional Importance of Alnus Pollen as an Aeroallergen: A Comparative Study of Alnus Pollen Counts from Worcester (UK) and Poznań (Poland)

Daily average Alnus pollen counts (1996-2005) from Worcester (UK) and Poznań (Poland) were examined with the aim of assessing the regional importance of Alnus pollen as an aeroallergen. The average number of Alnus pollen grains recorded annually at Poznań was more than 2.5 times that of Worcester. Furthermore, daily average Alnus pollen counts exceeded the thresholds of 100, 500 and 1,000 grains/m3 more times at Poznań than Worcester. Skin prick test results (1996-2005) and allergen-specific IgE(asIgE) measurements using the CAP (Pharmacia) system (2002-2005), were supplied by the Allergic Diseases Diagnostic Centre in Poznań. The annual number of positive skin prick tests to Alnus pollen allergens was significantly related (p<0.05) to seasonal variations in the magnitude of the Alnus pollen catch recorded at Poznań (r=0.70). The symptoms of patients with positive skin prick tests to Alnus pollen allergens were: 51% pollinosis, 43% atopic dermatitis, 4% asthma, 1% chronic urticaria and 1% eczema. On a scale of 0-6, 20.5% of patients examined for serum asIgE in relation to Alnus pollen allergens had asIgE measurements in classes 5 and 6. Alnus pollen is generally considered to be mildly allergenic. However, the amount of Alnus pollen released into the atmosphere in places such as Poznań may increase its impact on the population and make it one of the more important aeroallergens present.

Are the Birch Trees in Southern England a Source of Betula Pollen for North London?

Birch pollen is highly allergenic. Knowledge of daily variations, atmospheric transport and source areas of birch pollen is important for exposure studies and for warnings to the public, especially for large cities such as London. Our results show that broad-leaved forests with high birch tree densities are located to the south and west of London. Bi-hourly Betula pollen concentrations for all the days included in the study, and for all available days with high birch pollen counts (daily average birch pollen counts >80 grains/m3), show that, on average, there is a peak between 1400 hours and 1600 hours. Back-trajectory analysis showed that, on days with high birch pollen counts (n=60), 80% of air masses arriving at the time of peak diurnal birch pollen count approached North London from the south in a 180 degree arc from due east to due west. Detailed investigations of three Betula pollen episodes, with distinctly different diurnal patterns compared to the mean daily cycle, were used to illustrate how night-time maxima (2200–0400 hours) in Betula pollen counts could be the result of transport from distant sources or long transport times caused by slow moving air masses. We conclude that the Betula pollen recorded in North London could originate from sources found to the west and south of the city and not just trees within London itself. Possible sources outside the city include Continental Europe and the Betula trees within the broad-leaved forests of Southern England.

Modelling Atmospheric Concentrations of Ragweed Pollen From Local and Distant Sources

Pollen grains from the genus ragweed (Ambrosia spp.) are important aeroallergens. In Europe, the largest sources of atmospheric ragweed pollen are the Rhône Valley (France), parts of Northern Italy, the Pannonian Plain and Ukraine. Episodes of Long Distance Transport (LDT) of ragweed pollen from these centres can cover large parts of Europe and are predominantly studied using receptor based models (Smith et al., (2013) and references therein). The clinical impact of allergenic ragweed pollen arriving from distant sources remains unclear (Cecchi et al. 2010). Although a recent study has found the major allergens of ragweed in air samples collected in Poznań, Poland, during episodes of long-distance transport from the Pannonian Plain (Grewling et al. 2013). The source orientated models SILAM, DEHM, COSMO-Art, METRAS and ENVIRO-HIRLAM currently report having the capability of modelling atmospheric concentrations of pollen in Europe. The performance of such source-orientated models is strongly dependent on the quality of the emissions data, which is a focus of current research (e.g. Thibaudon et al. (2014)). The output from these models are important for warning allergy sufferers in areas polluted by ragweed, but could also be used to warn the public of ragweed pollen being transported into areas where the plant is not abundant. Areas outside of the main areas of ragweed infection that contain considerable local populations must, however, also include local scale models. These models can be used to predict local concentrations, even when LDT is not present. This concept of combined LDT and local scale calculations has been shown to be work for air pollutants and is considered usable for urban scale calculations of aeroallergens once urban scale maps of aeroallergen sources have been produced.

Risk of Exposure to Airborne Ambrosia Pollen from Local and Distant Sources in Europe – an Example from Denmark

Background. Ambrosia artemisiifolia L. is a noxious invasive alien species in Europe. It is an important aeroallergen and millions of people are exposed to its pollen. Objective. The main aim of this study is to show that atmospheric concentrations of Ambrosia pollen recorded in Denmark can be derived from local or more distant sources. Methods. This was achieved by using a combination of pollen measurements, air mass trajectory calculations using the HYPLIT model and mapping all known Ambrosia locations in Denmark and relating them to land cover types. Results. The annual pollen index recorded in Copenhagen during a 15-year period varied from a few pollen grains to more than 100. Since 2005, small quantities of Ambrosia pollen has been observed in the air every year. We have demonstrated, through a combination of Lagrangian back-trajectory calculations and atmospheric pollen measurements, that pollen arrived in Denmark via long-distance transport from centres of Ambrosia infection, such as the Pannonian Plain and Ukraine. Combining observations with results from a local scale dispersion model show that it is possible that Ambrosia pollen could be derived from local sources identified within Denmark. Conclusions. The high allergenic capacity of Ambrosia pollen means that only small amounts of pollen are relevant for allergy sufferers, and just a few plants will be sufficient to produce enough pollen to affect pollen allergy sufferers within a short distance from the source. It is necessary to adopt control measures to restrict Ambrosia numbers. Recommendations for the removal of all Ambrosia plants can effectively reduce the amount of local pollen, as long as the population of Ambrosia plants is small.

Mesoscale Atmospheric Transport of Ragweed Pollen Allergens from Infected to Uninfected Areas

Allergenic ragweed (Ambrosia spp.) pollen grains, after being released from anthers, can be dispersed by air masses far from their source. However, the action of air temperature,humidity and solar radiation on pollen grains in the atmosphere could impact on the ability of long distance transported (LDT) pollen to maintain allergenic potency. Here, we report that the major allergen of Ambrosia artemisiifolia pollen (Amb a 1) collected in ambient air during episodes of LDT still have immunoreactive properties. The amount of Amb a 1 found in LDT ragweed pollen grains was not constant and varied between episodes. In addition to allergens in pollen sized particles, we detected reactive Amb a 1 in subpollen sized respirable particles. These findings suggest that ragweed pollen grains have the potential to cause allergic reactions, not only in the heavily infested areas but, due to LDT episodes, also in the regions unaffected by ragweed populations.

Investigating and Predicting Atmospheric Concentrations of Allergenic Fungal Spores (Alternaria, Cladosporium, Didymella and Ganoderma)

Air quality is an increasing concern of the European Union, local authorities, scientists and most of all inhabitants that become more aware of the quality of the surrounding environment. Bioaerosols may be consisted of various elements, and the most important are pollen grains, fungal spores, bacteria, viruses. More than 100 genera of fungal spores have been identified as potential allergens that cause immunological response in susceptible individuals. Alternaria and Cladosporium have been recognised as the most important fungal species responsible for respiratory tract diseases, such as asthma, eczema, rhinitis and chronic sinusitis. While a lot of attention has been given to these fungal species, a limited number of studies can be found on Didymella and Ganoderma, although their allergenic properties were proved clinically. Monitoring of allergenic fungal spore concentration in the air is therefore very important, and in particular at densely populated areas like Worcester, UK. In this thesis a five year spore data set was presented, which was collected using a 7-day volumetric spore trap, analysed with the aid of light microscopy, statistical tests and geographic information system techniques. Although Kruskal-Wallis test detected statistically significant differences between annual concentrations of all examined fungal spore types, specific patterns in their distribution were also found. Alternaria spores were present in the air between mid-May/mid-June until September-October with peak occurring in August. Cladosporium sporulated between mid-May and October, with maximum concentration recorded in July. Didymella spores were seen from June/July up to September, while peaks were found in August. Ganoderma produced spores for 6 months (May-October), and maximum concentration could be found in September. With respect to diurnal fluctuations, Alternaria peaked between 22:00h and 23:00h, Cladosporium 13:00-15:00h, Didymella 04:00-05:00h and 22:00h-23:00h and Ganoderma from 03:00h to 06:00h. Spatial analysis showed that sources of all fungal species were located in England, and there was no evidence for a long distance transport from the continent. The maximum concentration of spores was found several hours delayed in comparison to the approximate time of the spore release from the crops. This was in agreement with diurnal profiles of the spore concentration recorded in Worcester, UK. Spores of Alternaria, Didymella and Ganoderma revealed a regional origin, in contrast to Cladosporium, which sources were situated locally. Hence, the weather conditions registered locally did not exhibit strong statistically significant correlations with fungal spore concentrations. This has had also an impact on the performance of the forecasting models. The best model was obtained for Cladosporium with 66% of the accuracy.