Simultaneous determination of mercury methylation and demethylation capacities of various sulfate-reducing bacteria using species-specific isotopic tracers.

The use of species-specific isotopic tracers for inorganic and methyl mercury has allowed the simultaneous determination of the methylation and demethylation potentials of pure culture of isolated sulfate-reducing (SR) bacterial strains using low Hg species concentration levels (7 µg/L (199)Hg(II), 1 µg/L Me(201)Hg). A major advantage of the method reported here is that it can be used to follow simultaneously both the degradation of the species added but also the formation of their degradation products and thus the determination during the same incubation of the specific methylation/demethylation yields and rate constants. Methylation/demethylation capacities and extents have been found to differ between the tested strains and the tested conditions. The methylating/demethylating capacities of bacteria appear to be strain specific. All the methylating strains were found to demethylate methylmercury (MeHg). The active mechanism responsible for Hg methylation appears directly dependent on the bacterial activity but is not dependent on the metabolism used by the tested bacteria (sulfate reduction, fermentation, or nitrate respiration). The results provide confirmation that SR strains contribute to MeHg demethylation under anoxic conditions, leading to Hg(II) as the end product, consistent with the oxidative degradation pathway. Kinetic experiments have allowed specific transformation rate constants to be addressed for the two reversible processes and the reactivity of each isotopic tracer to be compared. The differential reactivity highlighted the different steps involved in the two apparent processes (i.e., uptake plus internal transformation of mercury species). Methylation appears as the slowest process, mainly controlled by the assimilation of Hg(II), whereas demethylation is faster and not dependent on the MeHg concentration.