Pollen and fungal spore sampling and analysis. Statistical evaluations

Statistical evaluations of samples obtained from a Burkard seven-day recording volumetric pollen/spore trap were performed to determine the precision of the sampling and analysis procedures. The reproducibility of co-located traps was also investigated. The results showed that pollen grain transect counting was not significantly different, while fungal spore counting produced statistically different results. There was no statistical difference in the number of pollen and fungal spores counted between the co-located samplers. Reasons for the differences in the fungal spore counts are presented.     

Pollen and Mould Allergy in Southern Sardinia (Italy): Comparison of Skin-Test Frequencies and Air Sampling Data

A study was carried out to investigate the influence of atmospheric pollen and fungi in determining allergic diseases by comparing the frequency of skin reactions to air sampling data over a 6-year period.

48% of our population reacted to at least one of the pollen and fungal extracts used. Among pollen, Gramineae gave the most frequent positive reactions, followed by Parietaria, Olea and Compositae. The most common positive skin tests in fungus sensitive patients were by extracts of Alternaria, Cladosporium, Aspergillus and Candida

As for the aerobiological survey, the general trend of pollen and molds was similar during the sampling period. The annual pollen catch did not show remarkable differences during the years sampled, whereas the total fungal spore count was highest in 1988 and 1990.

A comparison between aerobiological and clinical data revealed a good degree of concordance between total pollen counts and positive skin test frequencies for Urticaceae, Gramineae and Oleaceae but not for Compositae (high positive skin reactions and very low counts) and Cupressaceae (high counts and few skin reactions).

A less marked correlation has been found between fungal spore counts and positive skin-test frequencies as compared to pollen. Spores such as Cladosporium, which are present in large number in the air, appear to be less sensitizing, while certain spore types (e.g. Alternaria), seem to be able to sensitize patients in spite of their low atmospheric concentrations.     

Fungal spores prevalent in the aerosol of the city of Caxias do Sul, Rio Grande do Sul, Brazil, over a 2-year period (2001–2002)

This study was carried out over a 2-year period (2001 and 2002) with the aim of identifying the fungal population in the aerosol of the Southern city of Caxias do Sul, RS, Brazil. Sampling was performed using Hirstȁ9s non-viable volumetric method. Our results show the presence of a large number of fungal spore types, a total of 41. Three groups were predominant: Deuteromycotina, Ascomycotina and Basidiomycotina. In 2001, Deuteromycotina taxa represented 44.61% of the total annual spore counts, with the largest concentration occurring in the fall (58,637 spores); in 2002, it represented 40.03% of the total annual spore counts, and the largest concentration was obtained in the summer (68,317 spores). Ascomycotina was present at the same level (24.5%) in both years of sampling, and the highest concentrations were found out in the summer (2001: 42,183 spores; 2002: 29,461 spores). Basidiomycotina represented 22.37% of the total annual spore counts in 2001, and 20.41% in 2002, with the largest concentrations found in the summer (2001: 35,988 spores; 2002: 30,212 spores). The most frequent fungi found during the study period were Cladosporium, Coprinus, Leptosphaeria, Aspergillus/Penicillium and Ganoderma. Permanent aerobiological monitoring would be necessary to detect associated environmental variations.     

Effects of weather conditions on selected airborne fungal spores in the southern part of the state of Enugu,Nigeria

A study of airborne fungal spore was carried out at nine locations in the southern part of the state of Enugu, Nigeria, from March 2005 to February 2006. The aim of the study was to ascertain the variations in selected fungal spore types at the sites owing to weather conditions. The variation in airborne fungal spores of 14 taxa was studied using modified Tauber pollen traps including Alternaria, Corynespora, Curvularia, Drechslera type, Endophragmiella, Botryodiplodia, Ganoderma, Gliomastrix, Nigrospora, Pithomyces, Spegazzinia, Sporidesmium, Tetraploa and Ustilago. The frequency of the spore types recorded showed considerable variation. The highest spore counts were recorded in July, June and October. The highest numbers of fungal spores were recorded during the rainy season (June–October) to early dry season (November–December). The peak of occurrence of most selected fungal spore types was July. The highest percentages of fungal spores were documented at the recording stations Mgbowo Junction, UNTH Ituku Ozalla and Oji River Express Junction. Spearman’s correlation analyses were performed for the monthly amounts of the fungal spore types and monthly meteorological factors. The numbers of Curvularia, Nigrospora and Sporidesmium was significantly correlated with relative humidity, while those of Endophragmiella, Pithomyces and Nigrospora were significantly correlated with temperature. A significant correlation was also found between the number of Nigrospora spores and light intensity and Sporidesmium spores and wind velocity. Relative humidity and temperature seem to be the most important weather conditions affecting the frequency of the selected spore types in the atmosphere.     

Fern spore and pollen airspora profile of Singapore

Monitoring atmospheric fern spore and pollen loads in Singapore was initiated in June 1990. Aside from the more numerous fungal spores, fern spores and pollen grains made up 6.2–8.6% and 4.4–5.4% of the total airspora sampled, respectively. The most frequently encountered fern spores, in descending order, were those of Nephrolepis auriculata, Dicranopteris linearis, Stenochlaena palustris, Asplenium nidus, Pteridium aquilinum, and Dicranopteris curranii. For pollen grains, the most frequently encountered, in descending order, were Elaeis guineensis, Casuarina equisetifolia, Acacia auriculiformis, Kyllingia polyphylla, Podocarpus, and Poaceae pollen grains. Seasonal patterns for individual fern spore or pollen types were discernible despite the relatively uniform tropical climate in Singapore. The fern spore and pollen calendar for the period 1991–1995 was compared to that of 2005–2006 as a follow-up study to keep abreast with the rapidly changing landscape of Singapore. Diurnal patterns showing a late morning to afternoon peak period were seen in fern spores, while the peak period was in the morning for pollen types studied with the exception to oil palm pollen. Additionally, association between fern spore and pollen counts and local meteorological conditions were also analyzed and found to be highly correlated. This study has thus identified the fern spores and pollen airspora components, and determined the calendars, as well as diurnal profiles of the Singapore airspora and provides invaluable information for allergy studies by highlighting the trigger sources present in the environment.     

Development and validation of a 5-day-ahead hay fever forecast for patients with grass-pollen-induced allergic rhinitis

One-third of the Dutch population suffers from allergic rhinitis, including hay fever. In this study, a 5-day-ahead hay fever forecast was developed and validated for grass pollen allergic patients in the Netherlands. Using multiple regression analysis, a two-step pollen and hay fever symptom prediction model was developed using actual and forecasted weather parameters, grass pollen data and patient symptom diaries. Therefore, 80 patients with a grass pollen allergy rated the severity of their hay fever symptoms during the grass pollen season in 2007 and 2008. First, a grass pollen forecast model was developed using the following predictors: (1) daily means of grass pollen counts of the previous 10 years; (2) grass pollen counts of the previous 2-week period of the current year; and (3) maximum, minimum and mean temperature (R ²?=?0.76). The second modeling step concerned the forecasting of hay fever symptom severity and included the following predictors: (1) forecasted grass pollen counts; (2) day number of the year; (3) moving average of the grass pollen counts of the previous 2 week-periods; and (4) maximum and mean temperatures (R ²?=?0.81). Since the daily hay fever forecast is reported in three categories (low-, medium- and high symptom risk), we assessed the agreement between the observed and the 1- to 5-day-ahead predicted risk categories by kappa, which ranged from 65 % to 77 %. These results indicate that a model based on forecasted temperature and grass pollen counts performs well in predicting symptoms of hay fever up to 5 days ahead.     

Considerations in the grouping of plant and fungal taxa for an epidemiologic study

Although exposure to airborne pollen grains and fungal spores has been implicated as a causative factor for acute exacerbation of asthma, the few epidemiologic studies that have attempted to evaluate the relationship between these bioaerosols and asthma have used only total counts (ignoring the relative importance of different taxa) or a few predominant pollen or spore types (ignoring less abundant but potentially relevant groups). This paper reports the development of hypothesis‐driven exposure metrics (based on known aeroallergen associations with allergic asthma and other hypersensitivity diseases, pollen allergen cross‐reactivity, and the presence of local sources in the city of Fresno, California, USA) for a 3.5 year epidemiologic study of childhood asthma. Outdoor regional and neighborhood concentrations of pollen and spores were measured using Hirst‐type, 7‐day samplers. Indoor and outdoor residential concentrations were measured at 84 selected homes with similar 24‐hour slit impactors. All pollen and spore concentrations were recorded in 2‐hour intervals to assist in understanding diurnal fluctuations in aeroallergen concentrations, identify exposures during the time periods that children are outdoors, and study interaction between aeroallergens and other air contaminants, which were the primary focus of the study. The 124 pollen taxa that were observed were reduced to 15 categories and the 66 fungal and algal taxa were reduced to five categories that will be used in microenvironmental models to generate individual daily exposure estimates for each of the 315 children. These new exposure metrics will allow examination of health effects for taxa traditionally associated with allergy and those with locally elevated concentrations in combination with exposures to other indoor and outdoor air contaminants.     

Comparison of airborne herb pollen types in Córdoba (Southwestern Spain) and Poznan (Western Poland)

This study sought to compare airborne pollen counts for a number of common herbaceous species (Plantago, Chenopodiaceae–Amaranthaceae, Rumex, and Urticaceae) in two cities with differing weather conditions, Córdoba (Southwestern Spain) and Poznan (Western Poland). Pollen seasons for these species were studied from 1995 to 2005. Aerobiological sampling was performed using a Hirst type 7-day spore trap, in accordance with the procedure developed by the European Aerobiology Network. A Spearman correlation test was used to test for correlations between meteorological parameters and daily airborne pollen counts. The Spearman correlation test and the Wilcoxon signed ranks test were also used to compare mean daily pollen counts for the two study sites. In Córdoba, the pollen season generally started around two months earlier than in Poznan, and also lasted longer. These findings were attributed to the presence of a larger number of species in Córdoba, with overlapping pollen seasons, and also to more favorable weather conditions. Trends in pollen season start dates were fairly stable over the study period, with a slight tendency to delayed onset in Córdoba and a modest advance in start date in Poznan. The pollen season end date also remained reasonably stable over the study, with only a slight tendency for the season to end earlier in Córdoba and later in Poznan. A clear trend towards declining annual pollen counts was recorded over the study period for all pollen types in both cities.     

First results of a study on airborne sporomorps in Budapest,Hungary

The first results of an aerobiological study, with the help of a Burkard spore trap in Hungary, are reported. A pollen calendar of 1989 is presented. Out of the total identified sporomorphs, the most relevant 16 types of pollen and 10 of fungal spores are reported. We have pointed out, that in Budapest Ambrosia and Artemisia pollen and some fungal spores are particularly responsible for the long lasting allergies to pollen and spores in the autumn. The pollen frequencies and the symptom scores of patients of the Pediatrics (Semmelweis Medical School, Budapest) showed a good correlation. Weekly reports are published in a newspaper and in television.     

Predicting the grass pollen count from meteorological data with regard to estimating the severity of hayfever symptoms in Melbourne (Australia)

In Melbourne, Australia, grass pollen is the predominant cause of hayfever in late spring and summer. The grass pollen season has been monitored in Melbourne, using a Burkard spore trap, for 13 years (1975–1981, 1985 and 1991–1997). Total counts for grass pollen were highly variable from one season to the next (approximately 1000 to >8000 grains/m³). The daily grass pollen counts also showed a high variability (0 to approximately 400 grains/m³). In this study, the grass pollen counts of the 13 years (12 grass pollen seasons, extending from October to January) have been compared with meteorological data in order to identify the conditions that can determine the daily amounts of grass pollen in the air. It was found that the seasonal total of grass pollen was directly correlated with the rainfall sum of the preceding 12 months (1 September–31 August): seasonal total of grass pollen (counts/m³)=18.161 × rainfall sum of the preceding 12 months (mm) −8541.5 (r _s=0.74,P<0.005,n=12). The daily amounts of grass pollen in the air were positively correlated with the corresponding daily average ambient temperatures (P<0.001). The daily amount of grass pollen which was to be expected with a certain daily average temperature was linked to the seasonal total of grass pollen: in years with high total grass pollen counts, a lower daily average temperature was required for a high daily pollen count than in years with low total grass pollen counts. As the concentration of airborne grass pollen determines the severity of hayfever in sensitive patients, an estimation of daily grass pollen counts can provide an indication of potential pollinosis symptoms. We compared daily grass pollen counts with the reported symptomatic responses of hayfever sufferers in November 1985 and found that hayfever symptoms were significantly correlated to the grass pollen counts (P<0.001 for nasal,P<0.005 for eye symptoms). Thus, a combination of meteorological information (i.e. rainfall and temperature) allows for an estimation of the potential daily pollinosis symptoms during the grass pollen season. Here we propose a symptom estimation chart, allowing a quick prediction of eye and nasal symptoms that are likely to occur as a result of variations in meteorological conditions, thus enabling both physicians and patients to take appropriate avoidance measures or therapy.     

A comparison of total pollen counts obtained 5.6 km apart

Total pollen counts obtained from a single pollen-sampling device are often broadcasted by the news media to assist allergy sufferers. In releasing these data, it is assumed that counts obtained in a single location represent pollen conditions throughout a large city. Volumetric pollen data collected at two locations in St. Paul, Minnesota, were used to test the suitability of this practice. A total of 122 pollen counts performed between April and October 1993 at each location were compared. Although the total number of pollen grains recovered during the study period by the two samplers was nearly equivalent, some monthly and daily differences were uncovered. Differences between the locations grew as pollen counts increased and were most pronounced for counts exceeding 100 pollen grains/m³. Despite the differences between the locations, this study demonstrated that pollen counts obtained with a single pollen-sampling device may be cautiously used to represent pollen conditions in a large community.     

An assessment of the air quality in indoor and outdoor air with reference to fungal spores and pollen grains in four working environments in Kerala, India

Qualitative and quantitative analyses of airborne fungal spores and pollen grains in four working environments (market, saw mill, poultry and cow sheds) in Thiruvananthapuram, the capital city of Kerala, India, were carried out for 2 years using Burkard Personal Slide Sampler and Andersen Two-Stage Sampler. Total spore concentration in these sites was always higher in indoor environments than in outdoor environments. Difference in concentration was not statistically significant in any of these work places except in saw mill (t test, p < 0.05). The highest spore concentration was recorded here followed by market, poultry and cow sheds. A total of 32 fungal spore types from indoor environments and 33 spore types from outdoor environments were recorded. Of them, 16 spore types were common to all the sites. Ameropsores, Cladosporium, other basidiospores, Ganoderma and Nigrospora were the dominant spore types in both indoor and outdoor environments. A total of 27 species of viable fungi from indoor and 24 species from outdoor environments were identified. Penicillium citrinum, Aspergillus flavus and Aspergillus niger were the most dominant viable fungi isolated. In contrast, total pollen concentration was always higher in outdoor environments than in indoor environments. Twenty-nine pollen types from indoor and 32 pollen types from outdoor were captured during the sampling. Poaceae, Cocos, Artocarpus, Amaranthus/Chenopodium and Tridax were the common and dominant pollen types observed in all the sites. Peak spore and pollen incidence were recorded during the late rainy and dry seasons (October–February) in both indoor and outdoor environments. The study revealed high prevalence of predominantly allergenic fungal spores and pollen grains in all the four work places. Workers/visitors are at potential risk of susceptibility to respiratory/allergic disorders.     

Comparative study of airborne pollen counts located in different areas of the city of Córdoba (south-western Spain)

Airborne pollen counts are mainly determined using a volumetric suction sampler based on the impact principle, that is, a Hirst-type spore trap. As a consequence of their volumetric nature, samplers detect pollen from a wide area, and therefore, a single sampler is frequently used to acquire information on airborne pollen counts for the whole city. The main goal of the present study was to compare airborne pollen counts at two sites located at opposite ends (south-west vs. north-east) of the southern Spanish city of Córdoba, to assess the advantages and disadvantages of using more than one sampler in the city. Also, a comparative study was carried out using two samplers at the same site, in order to confirm the efficiency of the samplers. Results revealed that data from one volumetric sampler—located within a city of medium size with uniform topography and vegetation conditions—are sufficient to establish monitoring of the main airborne pollen types, the pollen seasons involved and the timing of peak counts. For clinical studies, however, data on pollen counts in specific areas of the city may be of value, since pollen intensity may vary from one district to another, mainly in the case of ornamental plants with a local distribution inside the city. Comparison of data obtained by the two samplers running at the same site indicated that potential inter-site differences could not be attributed to differences in sampler efficiency.     

Alternative statistical methods for interpreting airborne Alder (<Emphasis Type="Italic">Alnus glutimosa</Emphasis> (L.) Gaertner) pollen concentrations

This paper reports on the behaviour of Alnus glutinosa (alder) pollen grains in the atmosphere of Ponferrada (León, NW Spain) from 1995 to 2006. The study, which sought to determine the effects of various weather-related parameters on Alnus pollen counts, was performed using a volumetric method. The main pollination period for this taxon is January-February. Alder pollen is one of the eight major airborne pollen allergens found in the study area. An analysis was made of the correlation between pollen counts and major weather-related parameters over each period. In general, the strongest positive correlation was with temperature, particularly maximum temperature. During each period, peak pollen counts occurred when the maximum temperature fell within the range 9 degrees C-14 degrees C. Finally, multivariate analysis showed that the parameter exerting the greatest influence was temperature, a finding confirmed by Spearman correlation tests. Principal components analysis suggested that periods with high pollen counts were characterised by high maximum temperature, low rainfall and an absolute humidity of around 6 g m(-3). Use of this type of analysis in conjunction with other methods is essential for obtaining an accurate record of pollen-count variations over a given period.     

Performance of the Coriolis air sampler,a high-volume aerosol-collection system for quantification of airborne spores and pollen grains

The Coriolis δ air sampler manufactured by Bertin Technologies (France) is a continuous air sampler, dedicated to outdoor monitoring of airborne spores and pollen grains. This high-volume sampler is based on patented Coriolis technology delivering a liquid sample. The air is drawn into a conical vial in a whirling type motion using suction; particles are pulled against the wall by centrifugal force. Airborne particles are separated from the air and collected in a liquid medium. This innovative solution allows rapid analysis by several techniques including PCR assay and serological assay in order to measure the antigenicity/allergenicity of pollen grains and fungal spores. Also, traditional counting of pollen grains or taxa identification by optical microscopy can be done. A study has been carried out by the Health Protection Agency (HPA), Porton Down, UK, to measure the physical efficiency of the Coriolis air sampler. The physical efficiency of the sampler for collection of micro-organism-laden particles of various sizes has been compared with that of membrane filter samplers using the techniques described by ISO 14698-1. The Coriolis was operated simultaneously with membrane filter samplers in a controlled room where they were challenged with uniform-sized particles of different diameters containing bacterial spores. For the larger particle sizes, it was found that the physical efficiency of the Coriolis was 92% for 10-μm particles. The biological performance of the Coriolis in the collection of airborne fungal spores and pollen grains was evaluated in comparison with a Hirst spore trap (one-week tape-on-drum type sampler) which is one of the most frequently used traps in the measurement of outdoor pollen grain concentrations. The advantages and limitations of both technologies are discussed. The Coriolis was operated simultaneously with a Hirst spore trap in the sampling station of Réseau National de Surveillance Aérobiologique, France (RNSA); the pollen grain and fungal spore counts were analysed by optical microscopy. The pollen grain count m⁻³ collected was compared for both devices. The dispersion values were obtained and statistical analysis was carried out. This study shows that the Coriolis air sampler provided equivalent recovery of pollen grain and fungal spores compared with the volumetric trap standard method (not significantly different, W test, α = 0.05). Nowadays, the French-led project, acronym MONALISA, with financial support from the European Commission––Life-Environment (LIFE05 ENV/F/000068), is testing this innovative air sampler in order to measure the antigenicity/allergenicity of the main aeroallergen particles, i.e. Betula (birch), Poaceae (grasses), Parietaria (pellitory), Olea spp (olive tree), and Artemisia (mugwort) pollen grains, and Alternaria (fungal spores) to validate a new approach of monitoring instead of quantifying pollen grains by their morphology. The robustness and efficiency of the MONALISA system is being demonstrated at a national level throughout Europe in eight different countries with different bio-climatic and topography characteristics: France, UK, Finland, Poland, Spain, Portugal, Switzerland, and Italy.     

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.     

Aerobiology of Urticaceae pollen in Trieste (NE Italy)

The aerobiological behaviour of Urticaceae in Trieste and the correlations with the meteorological parameters were examined. Airborne pollen was collected from 1990 to 1999 using a Hirst type spore trap (Burkard) and the data interpretation was performed according to the standard method adopted by the Italian Aeroallergen Network. The main pollen season of Urticaceae in Trieste goes from mid-April to mid-September. The highest values occur in May and June. Although different seasonal patterns are found every year, the main peak occurs on average at the beginning of May, followed by other decreasing peaks until September. Thecumulative counts vary greatly over the years, with a mean value of 18.315 p/m³. The maximum annual total pollen grains was registered in 1996 and the minimum in 1991. Spearman's correlation was used to establish the relationship between the daily pollen counts and the daily meteorological data both considering their original quantitative values and transformed values according to their day by day changes. Daily pollen concentrations present usually positive correlation with temperature, negative with rainfall and wind speed and no correlation with humidity. Better results were obtained with transformed values.     

Atmospheric mold spore counts in relation to meteorological parameters

 Fungal spore counts of Cladosporium, Alternaria, and Epicoccum were studied during 8 years in Denver, Colorado. Fungal spore counts were obtained daily during the pollinating season by a Rotorod sampler. Weather data were obtained from the National Climatic Data Center. Daily averages of temperature, relative humidity, daily precipitation, barometric pressure, and wind speed were studied. A time series analysis was performed on the data to mathematically model the spore counts in relation to weather parameters. Using SAS PROC ARIMA software, a regression analysis was performed, regressing the spore counts on the weather variables assuming an autoregressive moving average (ARMA) error structure. Cladosporium was found to be positively correlated (P<0.02) with average daily temperature, relative humidity, and negatively correlated with precipitation. Alternaria and Epicoccum did not show increased predictability with weather variables. A mathematical model was derived for Cladosporium spore counts using the annual seasonal cycle and significant weather variables. The model for Alternaria and Epicoccum incorporated the annual seasonal cycle. Fungal spore counts can be modeled by time series analysis and related to meteorological parameters controlling for seasonallity; this modeling can provide estimates of exposure to fungal aeroallergens. Received: 14 October 1996 / Revised: 17 February 1997 / Accepted: 28 February 1997     

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.