Thiothrix CT3 was isolated in pure culture by micromanipulation technique. The growth of this microorganism was analyzed in autotrophic, heterotrophic and mixotrophic conditions, in a bicarbonate-containing mineral medium supplemented with thiosulphate and/or acetate. Thiothrix CT3 was able to grow in all the conditions examined: the maximum growth rates estimated were 1.8, 2.5 and 2.5 d−1 respectively. The capacity of this organism to grow autotrophically, as supposed by Winogradsky in 1888, is a strong advantage over the other bacteria present in activated sludge, when sulphides are produced or carried by the sewer. Both the maximum growth rate and growth yield coefficient shown during heterotrophic and mixotrophic growth were comparable, indicating that the simultaneous presence of two substrates (acetate and a reduced sulphur compound) does not give it any particular advantage. The strong presence of Thiothrix spp. in many plants located in South Italy can be explained by the wide nutritional versatility of this filamentous bacterium.