Three strains of Mn-oxidizing fungi were isolated from manganese-rich aquatic environments: sediment in a stream (Komanoyu) in Mori-machi and inflow to an artificial wetland in Kaminokuni-cho, Hokkaido, Japan. The characteristics of each strain were then established. Genetic analysis based on the ribosomal RNA (rRNA) gene was performed to clarify their classification. The sequences of the 18S rRNA and internal transcribed spacer (ITS1)-5.8S rRNA-ITS2 genes showed that all three strains are Ascomycetes. Based on its morphology, it seems probable that the KY-1 strain from Mori-machi belongs to the genus Phoma or Ampelomyces. The phylogenetic analysis indicates that this strain belongs to Phoma rather than Ampelomyces. Morphological identification of WL-1 and WL-2 strains from Kaminokuni-cho was impossible because of the lack of a sexual stage and specific organs. Phylogenetic analysis of the sequence in the ITS1-5.8S rRNA-ITS2 gene suggests that the WL-1 strain corresponds to Paraconyothyrium sporulosum and that WL-2 also belongs to the genus Paraconiothyrium. Because the ability to oxidize Mn has not been evaluated for most species of Phoma or Paraconiothyrium (Coniothyrium), further study is needed to confirm the status of these three strains.

The vertical distributions of heavy metals and other elements in sediment core samples in a landfill of abandoned mine tailings were examined. Core samples were analyzed by XRF and XRD, and further examined by ICP-AES measurements after acid-sequential extraction with HCl, HF, and HNO 3 . According to XRD, the sediments contained α-quartz as the main component, and sericite, chlorite, rhodochrosite, pyrite, jarosite, sphalerite, and galena as minor components. A complete decomposition method using sequential extraction for the mine tailings sediments was established at ambient temperatures. Acid sequential extraction was useful for the analysis of minerals in mine tailings which are difficult to detect by XRD. Rhodochrosite and chlorite were dissolved by HCl-extraction, quartz and sericite by HF-extraction, and sulfides by HNO 3 -extraction. Rhodochrosite was the main source of high concentrations of Mn in mine drainage, highly distributed below the −100 cm zone in the mine tailings, combined with pyrite but not with sericite and quartz. The distributions of most heavy metals were related to those of pyrite.

The mechanism of surface water remediation in a natural wetland that is receiving heavy metal-rich acidic mine drainage was investigated. Selective sequential extraction was useful to derive the mechanisms of heavy metal removal in the wetland. In the upstream portion of the wetland, dissolved Fe was removed mainly as oxide-bounded mineral phases, such as hydroxides. These are important for the subsequent removal of other heavy metals. Other ion-exchangeable and carbonate-bounded heavy metals are also observed in the upstream, associated with Fe oxides. Organic matter and Fe-Mn oxides in the upstream remove Cu and Zn ions from the drainage, respectively. In the middle of portion of the wetland the removal of heavy metal ions in relatively low concentrations occurs by the emergent vegetation. Greater clay abundance and higher microbial activity of sulfate reducing bacteria in the downstream parts achieved low-level removal of metals. Multi-cell wetlands are recommended for the treatment of acidic metal bearing surface water drainage, if sufficient land area and expenses are available to construct.

Chemical transportation of heavy metals in the constructed wetland impacted by acid drainage was investigated seasonally using a combination of the selective sequential extraction of the sediments with elemental analysis of the emergent vegetation in the wetland. Manganese was dissolved from sediments in the constructed wetland by the contact with acid drainage, and then precipitated again as ionexchangeable forms. It was expected that a part of Mn and Fe bound to oxides were flown out of the wetland as suspended particulate matters. It was observed that there is passive absorption of Mn in leaves of Phragmites austlaris in the upstream of the wetland. The transportation of Cu clearly showed the seasonal variation: there was the decomposition of organic substances with high molecular weights by soil microorganisms in summer. Therefore, Cu was complexed to the humic substances, and dramatically adosorbed onto the roots of Phragmites austlaris in down stream of the wetland. It was also observed that there is active absorption of Fe in roots and leaves of Phragmites austlaris. Most of the zinc was strongly bounded to the sediments, therefore, scarcely uptaken to the vegetation. It was also found that there were heavy metal distributions between plant organs.

To apply manganese-oxidizing bacteria to waste water treatment, basic performance of the manganese-oxidizing bacterium, Leptothrix discophora, distributed by ATCC was examined. In addition, the effect of carbon fiber on the oxidation by the bacterium was also investigated, since carbon fiber was reported to accelerate the growth of activated sludge during sewage treatment. The bacterium was found to be active in the medium containing 24 ppm Mn(II) ions, the concentration being about 8 times higher than the recommended one and practically useful level. The oxidation rate was higher with the static culture than the shaking culture. This was considered to be due to physical damage to the sheath structure of bacteria which is reported to be important to oxidize Mn(II) ions. The carbon fiber did not accelerate the microbial oxidation of Mn(II) ions. This is partly attributed to the lack of contact between the bacterial cell, which floated as thin membranes, and the carbon fiber sunk in the bottom of vessels. However, oxidized Mn species precipitated on the carbon fiber, which resulted in an improvement of the transparency of water. This effect is different from the one caused by activated carbons, since the carbon fiber has very low specific surface area and no pore structure. The effect was more remarkable in the shaking culture than the static culture, indicating that organic substances originated from the bacterium play an important role in the adsorption of oxidized Mn species.