Soil, stream beds and cattle drinking troughs were sampled every 3 months over 3 years. More than 750 putative mycobacteria were isolated and grouped into more than 50 biotypes pending full identification. Samples from woodland and farmed land yielded fewer isolates per site compared with other terrains (P < 0.05). Some seasonal effects were noted but the greatest difference was between years 1 and 3. This appeared not to be due to differences in temperature, rainfall or experimental procedure, but coincided with the introduction of organic farming practices. In year 3 there was a significant increase in nitrate‐reducing slow growers, both pigmented (P≤ 0.006) and non‐pigmented strains (P≤ 0.002), and a shift in biotypes was noted. In contrast, all fast growers declined with time, as did those slow growers unable to reduce nitrate. Changing farming practice may alter the profile of environmental mycobacteria, which has important implications for the immunological priming of humans and animals.