*2.4. Determination of Bacterial Count and Activity of Soil Enzymes*

In soil samples, microbiological and biochemical analyses were carried out using standard methods, which are given in Table 3. These methods are described in detail in the manuscript of Borowik et al. [57] and Wyszkowska et al. [58]. Analyses were performed in four replications.

**Table 3.** Parameters for determining the number of organotrophic bacteria and actinobacteria, calculating colony development index and ecophysiological diversity index, and determining enzyme activity.


#### *2.5. Metagenomic Analysis*

Taxon of bacteria in soil samples was determined using analysis of the 16S rRNA encoding gene based on the hypervariable region V3–V4. Two primers were used for amplification: 1055F (5 -ATGGCTGTCGTCAGCT-3 ) and 1392R (5 -ACGGGCGGTGTGTAC-3 ). Polymerase chain reaction (PCR) was performed in real time in an Mx3000P thermocycler (Stratagene) and sequencing was in an Mx3000P thermocycler (Stratagene) at Genomed S.A. Warsaw, Poland.
