Cross-feeding interactions in short chain gaseous alkane-driven perchlorate and selenate reduction

Short chain gaseous alkanes (SCGAs) mainly consist of methane (CH₄), ethane (C₂H₆), propane (C₃H₈) and butane (C₄H₁₀). The first three SCGAs have been shown to remove perchlorate (ClO₄⁻) and selenate (SeO₄²⁻), yet it is unknown whether C₄H₁₀ is available to reduce these contaminants. This study demonstrated that C₄H₁₀ fed biofilms were capable of reducing ClO₄⁻ and SeO₄²⁻ to chloride (Cl⁻) and elemental selenium (Se⁰), respectively, by employing two independent membrane biofilms reactors (MBfRs). Batch tests showed that C₄H₁₀ and oxygen fed biofilms had much higher ClO₄⁻ and SeO₄²⁻ reduction rates and enhanced expression levels of bmoX and pcrA than that without C₄H₁₀ or O₂. Polyhydroxyalkanoates (PHA) accumulated in the biofilms when C₄H₁₀ was supplied, and they decomposed for driving ClO₄⁻ and SeO₄²⁻ reduction when C₄H₁₀ was absent. Moreover, we revisited the literature and found that a cross-feeding pathway seems to be universal in microaerobic SCGA-driven perchlorate and selenate reduction processes. In the ClO₄⁻-reducing MBfRs, Mycobacterium primarily conducts C₂H₆ and C₃H₈ oxidation in synergy with Dechloromonas who performs perchlorate reduction, while both Mycobacterium and Rhodococcus carried out C₄H₁₀ oxidation with perchlorate-respiring Azospira as the partner. In the SeO₄²⁻-reducing MBfRs, Mycobacterium oxidized C₂H₆ solely or oxidized C₃H₈ jointly with Rhodococcus, while Burkholderiaceae likely acted as the selenate-reducing bacterium. When C₄H₁₀ was supplied as the electron donor, both Mycobacterium and Rhodococcus conducted C₄H₁₀ oxidation in synergy with unknow selenate-reducing bacterium. Collectively, we confirm that from CH₄ to C₄H₁₀, all SCGAs could be utilized as electron donors for bio-reduction process. These findings offer insights into SCGA-driven bio-reduction processes, and are helpful in establishing SCGA-based technologies for groundwater remediation.