For acceleration of removing toxic metals from acid mine drainage (AMD), the effects of hydraulic retention time (HRT) and pH on the reactor performance and microbial community structure in the depth direction of a laboratory-scale packed-bed bioreactor containing rice bran as waste organic material were investigated. The HRT was shortened stepwise from 25 to 12 h, 8 h, and 6 to 5 h under the neutral condition using AMD neutralized with limestone (pH 6.3), and from 25 to 20 h, 12 h, and 8 to 7 h under the acid condition using AMD (pH 3.0). Under the neutral condition, the bioreactor stably operated up to 6 h HRT, which was shorter than under the acid condition (up to 20 h HRT). During stable sulfate reduction, both the organic matter-remaining condition and the low oxidation–reduction potential condition in lower parts of the reactor were observed. Principal coordinate analysis of Illumina sequencing data of 16S rRNA genes revealed a dynamic transition of the microbial communities at the boundary between stable and unstable operation in response to reductions in HRT. During stable operation under both the neutral and acid conditions, several fermentative operational taxonomic units (OTUs) from the phyla Firmicutes and Bacteroidetes dominated in lower parts of the bioreactor, suggesting that co-existence of these OTUs might lead to metabolic activation of sulfate-reducing bacteria. In contrast, during unstable operation at shorter HRTs, an OTU from the candidate phylum OP11 were found under both conditions. This study demonstrated that these microorganisms can be used to monitor the treatment of AMD, which suggests stable or deteriorated performance of the system.Electronic supplementary materialThe online version of this article (doi:10.1186/s13568-017-0440-z) contains supplementary material, which is available to authorized users.

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This study demonstrated heavy metal removal from neutral mine drainage of a closed mine in Kyoto prefecture in pilot-scale constructed wetlands (CWs). The CWs filled with loamy soil and limestone were unplanted or planted with cattails. The hydraulic retention time (HRT) in the CWs was shortened gradually from 3.8 days to 1.2 days during 3.5 months of operation. A short HRT of 1.2 days in the CWs was sufficient to achieve the effluent standard for Cd (0.03 mg/L). The unplanted and the cattail-planted CWs reduced the average concentrations of Cd from 0.031 to 0.01 and 0.005 mg/L, Zn from 0.52 to 0.14 and 0.08 mg/L, Cu from 0.07 to 0.04 and 0.03 mg/L, and As from 0.011 to 0.006 and 0.006 mg/L, respectively. Heavy metals were removed mainly by adsorption to the soil in both CWs. The biological concentration factors in cattails were over 2 for Cd, Zn, and Cu. The translocation factors of cattails for all metals were 0.5–0.81. Sulfate-reducing bacteria (SRB) belonging to Deltaproteobacteria were detected only from soil in the planted CW. Although cattails were a minor sink, the plants contributed to metal removal by rhizofiltration and incubation of SRB, possibly producing sulfide precipitates in the rhizosphere.

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