## 2 Materials and Methods  
### Soil  
Information on estrone soils was present in the literature, but more from a biogeochemical perspective [29], and even in these studies, the focus was more on soils with a much larger extension, such as thermozems and cambisols, and much less or nothing on less common soil groups. When the internet search was carried out for the current work on fluvisol, there was no information present (on Web of Science, search criteria 'estrogens AND fluvisol' and 'estrone AND fluvisol'). To address this knowledge gap, three soil groups from the Czech Republic used in agriculture were chosen for comparison: cambisol, chernozem, and fluvisol. Factors considered when choosing them were (1) how widespread they are in the Czech agriculture (cambisol is the most widespread soil type in the Czech Republic, chernozem is considered the best soil for such use, and fluvisol is the predominant soil in the riverine areas used in agriculture), and (2) their general soil properties, mainly (i) organic matter content (generally high in chernozem, low in cambisol and varying in fluvisols, depending on the quantity and quality of the alluvial sediments), (ii) texture and porosity (expected to be small for fluvisol as it can contain very fine sediments, and higher in chernozem and cambisol) and (iii) hydraulic properties (good drainage in chernozems and cambisols, and less in fluvisols) ([[https://www.fao.org/soils-portal/data-hub/soil-classification/world-reference-base/en/](https://www.fao.org/soils-portal/data-hub/soil-classification/world-reference-base/en/)]([https://www.fao.org/soils-portal/data-hub/soil-classification/world-reference-base/en/](https://www.fao.org/soils-portal/data-hub/soil-classification/world-reference-base/en/)), accessed on 22 October 2024). These factors influence estrone mobility and transformation,but also the structure of microbial communities in the soils and hence the estrone degradation ability.  
The soils were taken from the Czech University of Life Sciences Prague experimental fields from Suchdol, Humpolec, and Patek sites, air-dried, manually ground in a mortar and passed through a 2 mm plastic sieve before use. Soils characteristics are included in Table 1. pH was determined after extraction with 0.01 M CaCl\({}_{2}\) solution (1:2.5, w/v). Total C and N contents were determined by the CHNS Vario MACRO cube (Elementar Analysensysteme GmbH, Langenselbold, Germany) from air-dried samples, while the content of available nutrients (K, P, Ca, Mg, Mn, S, Zn, Fe, Cu, Cr) was estimated by the Mehlich III method [30] with a subsequent determination by inductively coupled plasma optical emission spectrometry (ICP-OES, Agilent 720, Agilent Technologies Inc., Santa Clara, CA, USA). The instrument was equipped with a two-channel peristaltic pump, a Struman-Masters spray chamber, and a V-groove pneumatic nebulizer made of inert material (experimental conditions: power of 1.2 kW, plasma flow of 15.0 L min\({}^{-1}\), auxillary flow of 0.75 L min\({}^{-1}\), nebulizer flow of 0.9 L min\({}^{-1}\)). Calibration curves were prepared as aqueous solutions of the stock solution acidified by 1.5% HNO\({}_{3}\) with the concentration range varying between 1 and 100 mg\(\cdot\)L\({}^{-1}\) for K, Ca, Mg, 0.5-50 mg\(\cdot\)L\({}^{-1}\) for S and P, and 0.01-1 mg\(\cdot\)L\({}^{-1}\) for Mn, Zn, Fe, Cu, and Cr. The limits of detection (LOD) were estimated at 0.1 mg\(\cdot\)L\({}^{-1}\) for P, 0.05 mg\(\cdot\)L\({}^{-1}\) for S, 0.005 mg\(\cdot\)L\({}^{-1}\) for Ca, Mg, Fe, Cu, 0.002 mg\(\cdot\)L\({}^{-1}\) for Zn, and 0.001 mg\(\cdot\)L\({}^{-1}\) for Mn. The limits of quantification (LOQ) were estimated at 0.33 mg\(\cdot\)L\({}^{-1}\) for P, 0.16 mg\(\cdot\)L\({}^{-1}\) for S, 0.017 mg\(\cdot\)L\({}^{-1}\) for Ca, Mg, Fe, Cu, 0.007 mg\(\cdot\)L\({}^{-1}\) for Zn, and 0.003 mg\(\cdot\)L\({}^{-1}\) for Mn.