Airborne pollen and fungi indoors: Evidence from primary schools in Lithuania.

The number of children suffering from respiratory allergies and asthma has been increasing worldwide and, hence, it is crucial to understand the burden of inhalant biological particles present in school facilities, where children spend one third of their life. From the perspective of indoor air quality, while there are numerous studies on outdoor bioaerosol exposure, there are still uncertainties regarding the diversity and deposition of airborne pollen and fungi indoors. When it comes to schools, there is limited research as to the potential bioaerosol exposure. Here we studied the indoor environment of public schools aiming to reveal whether primary schools of different sizes and at localities of different levels of urbanization may exhibit a variability in the biodiversity and abundance of particles of biological origin, which could pose a risk to child health. To achieve this, 11 schools were selected, located in a variety of environments, from downtown, to city centre-periphery, and to the suburbs. Fungal and pollen samples were collected from various surfaces in school classrooms and corridors, using passive air sampling and swab sampling. We demonstrated that fungi and pollen are detected in school premises during and after the vegetation season. The highest diversity of bioaerosols was found on the top of cabinets and windowsills, with Penicillium, Cladosporium and Acremonium being the most abundant indoors. The levels of fungi were higher in schools with more students. The diversity and amount of pollen in the spring were significantly higher than in samples collected in autumn. Our findings complemented existing evidence that bioaerosol measurements in schools (including kindergartens or informal education facilities) are vital. Hence, we here suggest that, in addition to monitoring air quality and bacterial levels indoors, fungi and pollen measurements have to be integrated in the existing regular biomonitoring campaigns so as to prevent exposure, increase awareness and manage efficiently allergic symptomatology.

Patterns of Phenolic Compounds in Betula and Pinus Pollen.

In this study, phenolic compounds and their antioxidant activity in the pollen of anemophilous Betula and Pinus were determined. Spectrophotometric, high-performance thin-layer and liquid chromatography methods were applied. Free phenolic compounds (free PC) and phenolic compounds bound to the cell wall (bound PC) were analysed in the pollen extracts. Regardless of the pollen species, their content was 20% higher than that in bound PC extracts. Pinus pollen extracts contained 2.5 times less phenolic compounds compared to Betula. Free PC extraction from the deeper layers of Pinus pollen was minimal; the same content of phenolic compounds was obtained in both types of extracts. The bioactivity of pollen (p < 0.05) is related to the content of phenolic compounds and flavonoids in Betula free PC and in bound PC, and only in free PC extracts of Pinus. Rutin, chlorogenic and trans-ferulic acids were characterised by antioxidant activity. Phenolic acids accounted for 70−94%, while rutin constituted 2−3% of the total amount in the extracts. One of the dominant phenolic acids was trans-ferulic acid in all the Betula and Pinus samples. The specific compounds were vanillic and chlorogenic acids of Betula pollen extracts, while Pinus extracts contained gallic acid. The data obtained for the phenolic profiles and antioxidant activity of Betula and Pinus pollen can be useful for modelling food chains in ecosystems.

Enrichment of Water Bodies with Phenolic Compounds Released from Betula and Pinus Pollen in Surface Water.

Betula and Pinus pollen, which are dispersed in natural surface waters, release biologically active compounds into the water bodies. This study aims to evaluate variations in the distribution and composition of phenolic compounds in suspended particles in natural water bodies during pollen spreading. Samples taken from water bodies of different trophic levels were analyzed by microscopy, UV/VIS spectroscopy, HPTLC, and HPLC/DAD. The study revealed that the total phenolic content in water-suspended particles varied from 3.0 mg/g to 11.0 mg/g during Betula and Pinus pollen spreading. It was also observed that the surface water of dystrophic natural lakes had a higher content of phenolic compounds than the eutrophic, hypereutrophic, and mesotrophic water bodies. Chlorogenic, trans-ferulic, vanillin, and 3,4-dihydroxybenzoic acids were frequently detected in the surface water samples. Experimental measurements have shown variations in the release of phenolic compounds from Betula pollen into water (p < 0.05). After the exhibition of pollen, the distilled water predominantly contained bioactive chlorogenic acid. Further in situ investigations are necessary to gain a more comprehensive understanding of the function of phenolic compounds in aquatic ecosystems. The exploration of the release of bioactive compounds from pollen could provide valuable insights into the potential nutritional value of pollen as a nutrient source for aquaculture.

Bioaerosols in the atmosphere at two sites in Northern Europe in spring 2021: Outline of an experimental campaign.

A coordinated observational and modelling campaign targeting biogenic aerosols in the air was performed during spring 2021 at two locations in Northern Europe: Helsinki (Finland) and Siauliai (Lithuania), approximately 500 km from each other in north-south direction. The campaign started on March 1, 2021 in Siauliai (12 March in Helsinki) and continued till mid-May in Siauliai (end of May in Helsinki), thus recording the transition of the atmospheric biogenic aerosols profile from winter to summer. The observations included a variety of samplers working on different principles. The core of the program was based on 2- and 2.4--hourly sampling in Helsinki and Siauliai, respectively, with sticky slides (Hirst 24-h trap in Helsinki, Rapid-E slides in Siauliai). The slides were subsequently processed extracting the DNA from the collected aerosols, which was further sequenced using the 3-rd generation sequencing technology. The core sampling was accompanied with daily and daytime sampling using standard filter collectors. The hourly aerosol concentrations at the Helsinki monitoring site were obtained with a Poleno flow cytometer, which could recognize some of the aerosol types. The sampling campaign was supported by numerical modelling. For every sample, SILAM model was applied to calculate its footprint and to predict anthropogenic and natural aerosol concentrations, at both observation sites. The first results confirmed the feasibility of the DNA collection by the applied techniques: all but one delivered sufficient amount of DNA for the following analysis, in over 40% of the cases sufficient for direct DNA sequencing without the PCR step. A substantial variability of the DNA yield has been noticed, generally not following the diurnal variations of the total-aerosol concentrations, which themselves showed variability not related to daytime. An expected upward trend of the biological material amount towards summer was observed but the day-to-day variability was large. The campaign DNA analysis produced the first high-resolution dataset of bioaerosol composition in the North-European spring. It also highlighted the deficiency of generic DNA databases in applications to atmospheric biota: about 40% of samples were not identified with standard bioinformatic methods.

Clustering approach for the analysis of the fluorescent bioaerosol collected by an automatic detector.

Automatically operating particle detection devices generate valuable data, but their use in routine aerobiology needs to be harmonized. The growing network of researchers using automatic pollen detectors has the challenge to develop new data processing systems, best suited for identification of pollen or spore from bioaerosol data obtained near-real-time. It is challenging to recognise all the particles in the atmospheric bioaerosol due to their diversity. In this study, we aimed to find the natural groupings of pollen data by using cluster analysis, with the intent to use these groupings for further interpretation of real-time bioaerosol measurements. The scattering and fluorescence data belonging to 29 types of pollen and spores were first acquired in the laboratory using Rapid-E automatic particle detector. Neural networks were used for primary data processing, and the resulting feature vectors were clustered for scattering and fluorescence modality. Scattering clusters results showed that pollen of the same plant taxa associates with the different clusters corresponding to particle shape and size properties. According to fluorescence clusters, pollen grouping highlighted the possibility to differentiate Dactylis and Secale genera in the Poaceae family. Fluorescent clusters played a more important role than scattering for separating unidentified fluorescent particles from tested pollen. The proposed clustering method aids in reducing the number of false-positive errors.

Genetic Loci Associated with Allergic Sensitization in Lithuanians.

Allergic rhinitis (AR) is a common and complex disease. It is associated with environmental as well as genetic factors. Three recent genome-wide association studies (GWAS) reported altogether 47 single nucleotide polymorphisms (SNPs) associated with AR or allergic sensitization (AS) in Europeans and North Americans. Two follow up studies in Swedish and Chinese replicated 15 associations. In these studies individuals were selected based on the self-reported AR, or AR/AS diagnosed using blood IgE test or skin prick test (SPT), which were performed often without restriction to specific allergens. Here we performed third replication study in Lithuanians. We used SPT and carefully selected set of allergens prevalent in Lithuania, as well as Illumina Core Exome chip for SNP detection. We genotyped 270 SPT-positive individuals (137 Betulaceae -, 174 Poaceae-, 199 Artemisia-, 70 Helianthus-, 22 Alternaria-, 22 Cladosporium-, 140 mites-, 95 cat- and 97 dog dander-sensitive cases) and 162 SPT-negative controls. We found altogether 13 known SNPs associated with AS (p ≤0.05). Three SNPs were found in Lithuanians sensitive to several allergens, and 10 SNPs were found in Lithuanians sensitive to a certain allergen. For the first time, SNP rs7775228:C was associated with patient sensitivity to dog allergens (F_A=0,269, F_U=0.180, P=0.008). Thus, careful assessment of AS allowed us to detect known genetic variants associated with AS/AR in relatively small cohort of Lithuanians.