Contamination of groundwater with chlorinated aliphatic hydrocarbons (CAHs) has become increasingly widespread. Enhanced reductive dechlorination (ERD) by emulsified vegetable oil (EVO) is an environmentally friendly, low-carbon and sustainable technology that effectively eliminates the threat from CAHs. To improve the permeability and bioavailability of EVO in the low-permeability substratum, nano-emulsified vegetable oil (nanoEVO) can be synthesized using a modified phase inversion composition method (MPIC) by increasing the emulsification temperature that reduces the shear force required for the formation of nanodroplets and the interfacial tension between the mixed phase and water. The emulsification temperature of 60 °C, the water content of 40–70%, the droplet diameter (DD) of nanoEVO prepared by MPIC is ∼200 nm, and had good monodispersion (≤0.2). The high molecular weight and low solubility of vegetable oil, and similar size of droplet make nanoEVO remains homogeneous after storage for > one year. NanoEVO can effectively biostimulate indigenous bacteria in CAHs-contaminated groundwater to dechlorinate 1,1,2-trichloroethane (1,1,2-TCA) to ethene via vinyl chloride (VC). The maximum dihaloelimination rate of 1,1,2-TCA was ∼66.06 μM·Cl−·day−1 and the maximum hydrogenolysis rate of VC was ∼35.67 μM·Cl−·day−1. NanoEVO effectively increased the relative abundance of Dehalococcoides from <0.10 to 9.61%. The abundances of Dhc 16S rRNA gene and dehalogenase gene vcrA increased by 102 and 103, respectively. Biostimulation shaped the structure of microbial communities and promoted hydrolytic acidification and anaerobic fermentation. Moreover, the syntrophic VFA-oxidizing bacteria participated in secondary fermentation and were closely correlated with H2 consumption metabolism. By providing carbon sources and maintaining an anaerobic environment with stable pH and ORP, nanoEVO biostimulated the hydrolytic acidification led by fermentative bacteria, secondary fermentation with syntrophic VFA-oxidizing bacteria and reductive dechlorination carried out by dechlorinator, thus greatly increasing the CAH-dechlorinating ability of indigenous microbial communities.
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