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Biodiversity assessment through DNA metabarcodingOur analysis detected 160 Operational Taxonomic Units (OTUs) with 12,007,712 sequenced reads, 222,370 ± 41,954 (sd) reads per sample, for a total of 54 sequenced samples. The rarefaction curves showed good sequencing effort for the samples (Supplementary Figure S1) which were rarefied to the least count among samples corresponding to 135,443 reads. Twenty OTUs, (7.2% of the total), were assigned to taxa not relevant to our work (mainly to mosses and ferns during the periods October 2014–March 2015 and July–October 2015). From the remaining OTUs, 108 (88% of the reads) were taxonomically assigned to 32 families of vascular plants (68 identified taxa) (Table 2, Supplementary Table S1) and 32 OTUs (4.8% of the reads) remained unidentified either because of low sequence identity and/or query coverage percentage or the absence of any sequence classification result, even when compared to the complete ‘Nucleotide’ Genbank database. The results of the taxonomic assignment to vascular plants are presented in Supplementary Table S1. The OTU sequences were assigned to plant taxa with at least 95% identity and coverage, from which 70% of the OTUs had ≥ 98% sequence identity with the assigned taxa. The positive control of the DNA extraction, Corylus avellana pollen, was correctly identified after HTS. From the 19 negative controls included in the extraction plate, one negative control was selected for sequencing, the only one with sufficient amplicon concentration (2 ng μl−1). In this sample two OTUs were detected (263,649 reads), both assigned to Quercus spp. and contributing  More

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The research was carried out on the basis of direct measurements in the surroundings of four selected working coal-fired power plants and four working coking plants. The samples of suspended dust PM10, respirable fraction PM2.5 and submicron particulate matter PM1 were collected in the surroundings of power generation facilities and in the surroundings of coking plants.Location of measurement pointsThe location of the measurement points was selected in southern Poland, around the selected four working coal-fired power plants and four working coking plants. The sampling points in the surroundings of the power plant (P1, P2, P3 and P4) and the coking plant (K1, K2, K3 and K4) were located at the distance of approximately 2 km to the north-east from the respective object (Fig. 1).Figure 1Location of the sampling sites (the map was generated based on data from the BDL18 website).Full size imageThe location of the measurement points was a compromise, taking into account the representativeness of the receptor, the possibility to connect the testing equipment and the consent of the property owners. To eliminate the impact of a heating season, and especially that of low emissions, presented in the studies by19, the measurement sessions were carried out only in the summer season. The samples of particulate matter were collected on a weekly basis, with 4 sessions at one site. The methodology applied in this work is presented in20,21. The location of measurement sites:

point P1: 50° 08′ 37.87″ N; 18° 32′ 15.76″ (Golejów—a suburban district of Rybnik in the Śląskie Voivodeship, in the vicinity of a working power plant with a capacity of 1775 MW; population:

2 300);

point P2: 50° 45′ 35.41″ N; 17° 56′ 20.43″ E (Świerkle—a rural area in the Opolskie Voivodeship (Dobrzeń Wielki commune) near a working power plant with a capacity of 1,492 MW; population: 520);

point P3: 50° 12′ 33.46″ N; 19° 28′ 28.77″ E (Czyżówka—rural area in the Małopolskie Voivodeship (commune of Trzebinia) near a working power plant with a capacity of 786 MW; population: 700);

point P4: 50° 13′ 48.90″ N; 19° 13′ 24.45″ E (suburbs of Jaworzno (Śląskie Voivodeship) in the vicinity of a 1,345 MW power plant; number of inhabitants: 95 500);

K1 point: 50° 10′ 11.36″ N; 18° 40′ 34.35″ E (Czerwionka—Leszczyny in the Śląskie Voivodeship, in the vicinity of a small coking plant; number of inhabitants: 27 300);

K2 point: 50° 3′ 19.76″ N; 18° 30′ 21.69″ E (Popielów—a suburban district of Rybnik in the Śląskie Voivodeship, surrounded by a small working coking plant; population:3 300);

K3 point: 50° 21′ 24.08″ N; 19° 21′ 37.46″ E (Łęka—Dąbrowa Górnicza district, in the Śląskie Voivodeship, surrounded by a large coking plant; number of inhabitants: 700);

K4 point: 50° 21′ 0.47″ N; 18° 53′ 15.44″ E (Bytom—a city in the Śląskie Voivodeship, a small coking plant located on the outskirts of the city; population: 174 700).

The state of air pollution with particulate matter in the area investigated in the study is affected by various local sources of pollution emissions. At the measurement sites P1, P2, P3 and P4, the emissions are mainly from power plant chimneys, but also from auxiliary processes, i.e. coal storage and its transport. In addition, the recorded emissions are also influenced by other industrial plants operating in the vicinity of the measurement sites, domestic and municipal sector and the impact of automotive industry. The measurement sites K1, K2, K3 and K4 involve primarily the emissions accompanying the processes of coal coking as well as auxiliary processes, i.e. coal deposition, its transmission, management of products and post-production wastes. Additionally, they are affected by the emissions from industrial plants and low emission sources operating in this area, as well as the emission from the combustion of solid fuels for domestic or municipal purposes, as well as by the automotive industry.Sampling processThe samples of suspended dust (PM10), respirable fraction (PM2.5) and submicron particulate matter (PM1) were collected using the Dekati PM10 cascade impactor serial No. 6648 by Dekati (Finland) with the air flow rate of (1.8 {mathrm{m}}^{3}/mathrm{h}). The impactor Dekati PM10 guarantees the collection of dust samples for three cutpoint diameters: 10 μm, 2.5 μm and 1 μm. For the sampling at the first, second and third stages of the impactor, polycarbonate filters were used (Nuclepore 800 203, with the diameter of 25 mm, by Whatman International Ltd., Maidstone, UK). At the fourth stage, the dust was collected on a Teflon filter for particles ≤ 1 μm in diameter (Pall Teflo R2PJ047, 47 mm in diameter, by Pall International Ltd., New York, NY, USA). The average volume of air passing through the filters was approximately 300 m3. The impactor’s capture efficiency was characterized by the uncertainty below 2.8%. The mass of dust collected at the individual stages of the impactor was determined by the gravimetric method, and it was referenced to the volume of passed air (left(mathrm{mu g}/{mathrm{m}}^{3}right)) according to the PN-EN1234122. All impactor samples were analysed by inductively coupled plasma mass spectrometry (ICP-MS).The samples were collected at a height of 1.5 m from the ground, i.e. in the breathing zone for people. The respective dust fractions were collected in 7-day cycles from 28 May to 24 September 2014 (16 weeks) in the surroundings of four working coal-fired power plants and from 4 May to 28 August 2015 (16 weeks) in the surroundings of four working coking plants. The measurement campaign comprised four measurement sessions separately for each sampling site. One session comprised dust sampling at each stage of the Dekati PM10 cascade impactor and filters used for reference. The filters were taken back after study period and labeled during the collection process in the field and stored in the plastic containers for safe transportation and storage in laboratory for further analysis.In each measurement session, blind filters were stored at the sampling site, but they were not subjected to exposure. The sample data were corrected from these blanks. The length of the measurement cycles was conditioned by the need to collect an appropriate amount of research material (with the aerodynamic diameter of the dust grains  10 μm). Analogous (7-day) periods of dust sampling were used in the studies by4,23.Polycarbonate and Teflon filters were conditioned before and after dust collection at a temperature of 20 ± 1 °C (relative humidity 50%(pm ) 5%) for 48 h, and then weighed on a microbalance with an accuracy of 1 (mathrm{mu g}) (MXA5/1, by RADWAG, Poland).Taking into account the measurement sessions at four sites in the surroundings of the power plant (P1 (div) P4) and at four sites in the surroundings of the coking plant (K1 (div) K4), the aggregate number of samples exceeded 450.Chemical analysisThe qualitative and quantitative analysis of the obtained solutions was performed by inductively coupled plasma mass spectrometry using an ICP-MS instrument (NexION 300D, PerkinElmer, Inc., Waltham, MA, USA). For all elements determined simultaneously, the same parameters of the instrument were used, which are presented in the publications20,21,24.As standards for the determination of 75As, 111Cd, 59Co, 53Cr, 200Hg, 55Mn, 60Ni, 206Pb, 121Sb and 82Se, we applied the 1000 (mathrm{mu g}/{mathrm{cm}}^{3}) CertPUR ICP multi-element standard solution VI for ICP-MS by Merck, Germany. Ten repetitions were performed for all samples. The determined limits of detection (LOD) were based on 10 independent measurements for blank test. For the results obtained in that way, the mean value and the value of the standard deviation SD were calculated. The values of LOD for individual elements were determined on the basis of the dependence (1):$$mathrm{LOD}= {mathrm{x}}_{mathrm{sr}}+ 3mathrm{SD}$$
(1)

where: xśr—mean concentration value of the element, (mathrm{g}/{mathrm{dm}}^{3}), SD—standard deviation.The determination correctness of the content of the elements was verified with the use of certified reference materials: European Reference Material ERM-CZ120 and Standard Reference Material SRM 1648a (National Institute of Standards and Technology, USA). The recovery with the use of the said certified reference materials was respectively as follows: As (111% for ERM-CZ120 and 96% for SRM 1648a), Cd (97% and 105%), Co (108% and 97%), Cr (103% and 94%), Mn (106% and 100%), Ni (107% and 102%), Pb (107% and 105%) and Sb (99% and 91%). The certified reference materials did not contain Hg or Se. More

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