The aim of the study was to demonstrate the impact of soil agricultural usage on the abundance of ammonifying bacteria (AB) and their activity, expressed as arginine ammonification (AA). Five agriculturally exploited types of soils (FAO): Haplic Luvisol, Brunic Arenosol, Mollic Gleysol, Eutric Fluvisol, and Rendzina Leptosol were studied. The controls were non-agricultural soils of the same type located in close proximity to agricultural sites. The tested soils varied in terms of pH (4.18–7.08), total carbon (8.39–34.90 g kg−1), easily degradable carbon content (0.46–1.11 g kg−1), moisture (5.20–13.50 %), and nitrogen forms (mg kg−1): 1.68–27.17, 0.036–0.862, 0.012–3.389 for nitrate nitrogen, nitrite nitrogen, and ammonia nitrogen, respectively. The AB abundance in agricultural soils ranged from 1.1 to 6.4 × 104 cfu g−1, while in the controls it was significantly higher—from 2.0 to 110 × 104 cfu g−1 of soil. Also, AA in the controls was three-times higher than in the agricultural soils. Strong associations between AA and the abundance of AB in the control (r = 0.954***) and agricultural soils (r = 0.833***) were proved. In the agricultural soils, the AB abundance and AA were influenced by pH (r = 0.746*** and r = 0.520***) and carbon content (r = 0.488*** and r = 0.391***). The AB abundance was also affected by easily degradable carbon (r = 0.517**) and nitrite nitrogen (r = 0.376*), whilst ammonium nitrogen influenced AA (r = 0.451*). Our results indicate that the abundance of AB and AA may be good indicators of soil biological conditions.