Dual C-Cl Isotope Analysis for Characterizing the Reductive Dechlorination of α- and γ-Hexachlorocyclohexane by Two Dehalococcoides mccartyi Strains and an Enrichment Culture

Abstract

Hexachlorocyclohexanes (HCHs) are persistent organic contaminants that threaten human health. Microbial reductive dehalogenation is one of the most important attenuation processes in contaminated environments. This study investigated carbon and chlorine isotope fractionation of α- and γ-HCH during the reductive dehalogenation by three anaerobic cultures. The presence of tetrachlorocyclohexene (TeCCH) indicated that reductive dichloroelimination was the first step of bond cleavage. Isotope enrichment factors (ε_C and ε_Cl) were derived from the transformation of γ-HCH (ε_C, from -4.0 ± 0.5 to -4.4 ± 0.6 ‰; ε_Cl, from -2.9 ± 0.4 to -3.3 ± 0.4 ‰) and α-HCH (ε_C, from -2.4 ± 0.2 to -3.0 ± 0.4 ‰; ε_Cl, from -1.4 ± 0.3 to -1.8 ± 0.2 ‰). During α-HCH transformation, no enantioselectivity was observed, and similar ε_c values were obtained for both enantiomers. The correlation of ¹³C and ³⁷Cl fractionation (Λ = Δδ¹³C/Δδ³⁷Cl ≈ ε_C/ε_Cl) of γ-HCH (from 1.1 ± 0.3 to 1.2 ± 0.1) indicates similar bond cleavage during the reductive dichloroelimination by the three cultures, similar to α-HCH (1.7 ± 0.2 to 2.0 ± 0.3). The different isotope fractionation patterns during reductive dichloroelimination and dehydrochlorination indicates that dual-element stable isotope analysis can potentially be used to evaluate HCH transformation pathways at contaminated field sites.