3.2.1. Area A

MTC was determined in each of the tested samples around the farm, regardless of the sampling season (except point 6A\_J). However, the concentrations in the samples were significantly lower—ranging from 4.69 × 10−<sup>3</sup> ± 0.16 ng/g d.w in 7A\_M to 7.08×10−<sup>4</sup> ± 0.263 ng/g d.w in 5A\_M. Similarly, SMZ was present in eight samples, and was determined in relatively high concentration levels. The range was from 68.70 ± 1.74 ng/g d.w. in 3A\_J, to 0.81 ± 0.19 ng/g d.w in 2A\_M. SA was detected in a range from 157.30 ± 4.65 ng/g d.w in 7A\_M, to 27.19 ± 6.01 ng/g d.w in 3A\_J. CBZ was found in four samples, three of which were collected in March. The highest concentration of CBZ was determined in sample 7A\_M and amounted to 1.96 ± 0.17 ng/g d.w. In turn, the lowest concentration of CBZ was determined in the July 4A\_J sample, with concentration equal to 0.70 ± 0.26 ng/g d.w. TMP was detected only in two samples from March on a similar level (7A\_M 0.80 ± 0.03 ng/g d.w, 5A\_M 0.75 ± 0.02 ng/g d.w.). The other four determined pharmaceuticals were found in individual soil samples. NIM was determined in the 4A\_M sample in a concentration level of 3.43 ± 0.03 ng/g d.w., MET in the 1A\_M sample at a concentration level of 2.99 ± 0.04 ng/g d.w., and PROP in sample 7A\_M at a concentration level equal to 0.97 ± 0.005 ng/g d.w.

#### 3.2.2. Areas B and C

As was mentioned, three other pharmaceuticals were found in the reference samples: CAF, PAR, and SNA. CAF (0.70 ± 0.13 ng/g d.w.) and PAR (11.31 ± 0.29 ng/g d.w.) were determined only in sample 1B\_M. In turn, SNA was determined in all three reference samples with the highest concentration (3518.22 ± 146.05 ng/g d.w.) in sample 1B\_J. In sample 1B\_M, which is the winter counterpart of 1B\_J, the concentration of SNA was 848.83 ± 1.19 ng/g d.w. In sample 1C\_J, SNA was determined in the concentration level of 746.57 ± 3.90 ng/g d.w.

#### *3.3. Analysis of Microbial Resistance in Soil Samples*

The bacterial communities able to grow from soil samples collected around the poultry breeding building were shown to be diverse (Supplementary Materials Figure S1). From each collecting time point, a few differently-looking bacterial colonies were picked and further analyzed for antibiotic resistance. Microbial resistance of selected soil bacterial isolates was performed for five different antibiotics, two of which (TMP and SMZ) were determined in soil samples in this study. Three other antibiotics (TET, SPEC and CIP) were analyzed since they prove to be the ones that are commonly used in chicken treatment during their growth in intensive breeding farms [23,24]. The average inhibition zones for bacterial strains isolated from different sampling points situated in zones A, B and, C around the poultry breeding farm are presented in Table 3. The least susceptibility of the tested strains was observed for sulfametazine and trimethoprim, where only six and seven susceptible isolates were found in chicken farm soil, respectively. In the soil sample collected in zone B, this number was even lower, as only two strains were susceptible to TMP and none was susceptible to SMZ. In zone C, none of the strains was susceptible to TMP and SMZ; however, here the number of tested strains was only four. On the other hand, the highest susceptibility was shown for CIP (all strains from each type of soil were susceptible), and an intermediate one was shown for TET and SPEC. Taking into account the results obtained for 85 random bacterial isolates, coming from different areas around the poultry breeding building, we found no significant difference between the bacterial resistance tested for different areas (Table 3).

**Table 3.** Antibiotic resistance of microbial strains isolated from soil collected in three different zones around the poultry farm building. A (0–50 m around the poultry farm building), B (approx. 500 m from poultry farm building), and C (approx. 1000 m from the poultry farm building). Inhibition zones around different antibiotics: trimethoprim (TMP) 5 μg, sulfametazine (SMZ) 20 μg, tetracycline (TET) 10 μg, spectinomycin (SPEC) 25 μg, and ciprofloxacin (CIP) 10 μg, are presented as average diameter (mm) of inhibition zone with standard deviation for different soil sampling zones.

