The oxidation activity of molecular tritium (HT) in soils and vegetation collected in experimental plots during the 1994 chronic HT release experiment at Chalk River was determined in in vitro laboratory experiments after the release. HT oxidation activity was highest in surface soils in the natural plot, about 3–4 times that in soils in the cultivated plots. HT oxidation activity in weeds and Komatsuna leaves was about 2 and 0.4 % of that in the cultivated soil, respectively. The number of HT-oxidizing bacteria isolated from soils was highest in the surface soil (0–5 cm) in the natural plot. The viable cell numbers in surface soils in the cultivated and natural plots were almost the same. The total occurrence rates of HT-oxidizing bacteria in the surface soils were 22 % in the natural plot, and 7.5 % in the cultivated plot. The occurrence rates of HT-oxidizing airborne bacteria during the release on two culture media were 4.2 and 1.9 %.