Characteristics of attachment and growth of Thiobacillus caldus on sulphide minerals: a chemotactic response to sulphur minerals?

Abstract

To further our understanding of the ecological role of sulphur‐oxidizing microorganisms in the generation of acid mine drainage (AMD), growth and attachment of the chemoautotrophic sulphur‐oxidizing bacterium, Thiobacillus caldus, on the sulphide minerals pyrite, marcasite and arsenopyrite was studied. Growth curves were estimated based on total cells detected in the system (in suspension and attached to mineral surfaces). In general, higher cell numbers were detected on surfaces than in suspension. Fluorescent in situ hybridizations to cells on surfaces at mid‐log growth confirmed that cells on surfaces were metabolically active. Total cell (both surface and solution phase) generation times on pyrite and marcasite (both FeS₂) were calculated to be ≈ 7 and 6 h respectively. When grown on pyrite (not marcasite), the number of T. caldus cells in the solution phase decreased, while the total number of cells (both surface and solution) increased. Additionally, marcasite supported about three times more total cells (≈ 3 × 10⁹) than pyrite (≈ 8 × 10⁸). This may be attributed to the dissolution rate of marcasite, which is twice that of pyrite. Epifluorescent and scanning electron microscopy (SEM) were used to analyse the cell orientation on surfaces. Results of Fourier transform analysis of fluorescent images confirmed that attachment to all three sulphides occurred in an oriented manner. Results from high‐resolution SEM imaging showed that cell orientation coincides with dissolution pit edges and secondary sulphur minerals that develop during dissolution. Preferential colonization of surfaces relative to solution and oriented cell attachment on these sulphide surfaces suggest that T. caldus may chemotactically select the optimal site for chemoautotrophic growth on sulphur (i.e. the mineral surface).