Bacillus species is an important microbial species in the activated sludge process. Some researchers reported that the predominance of Bacillus spp. improves treatment performance and good sludge settleability. The viable count of Bacillus spp. is usually measured by the plate culture method. Recently developed massive sequence technology has been applied to activated sludge samples; this technique provides more detailed information on its microbial community. However, the relationship between the number of Bacillus spp. and microbial compositions is not yet well understood. In order to elucidate this relationship, microbial community analyses based on 16S rRNA gene sequence and cell count of Bacillus spp. were conducted. Activated sludge samples, including those from conventional activated sludge, sequencing batch, and oxidation ditch processes, were collected and subjected to analysis. The results of microbial community analysis revealed that Proteobacteria and Bacteroides were the predominant bacterial phyla and overall community compositions resembled each other at the phylum level. The detection ratio for Bacillus spp. was 0-0.33%, and the number of Bacillus spp. ranged from 10 5 to more than 10 8 colonies/g-MLSS. The results showed that the number of Bacillus spp. and detection ratio showed a similar trend, and thus, these analyses could be complementary to each other.

Nitrification and denitrification processes occur simultaneously in aerobic wastewater biofilms. Although wide regions of the world have average temperatures of less than 15 °C for a half year, few studies have investigated the nitrogen removal by nitrification and denitrification in a single-stage aerobic biofilm reactor used for treating real wastewater under low-temperature conditions. This study showed successful wastewater treatment in a high average nitrogen removal rate of 78% at low water temperatures by simultaneous nitrification and denitrification in a rotating biological contactor (RBC) biofilm. Batch operations using the RBC to evaluate the rates of ammonium decrease at low temperatures demonstrated that the rate of ammonium decrease at 8 °C was 76% of that at 20 °C. Daily monitoring of nitrification and denitrification rates suggested that the denitrification rate was highly correlated with the nitrification rate. Next-generation sequencing (NGS) analysis revealed the presence of diverse and abundant denitrifying bacteria and aerobic bacteria in the RBC biofilm more than those in the activated sludge samples, which probably enabled the achievement of the high nitrogen removal rates at such low temperatures. Furthermore, correlation with the colony counts showed that the NGS analysis had the quantification range of three orders of magnitude (from 0.001% to 1%).

The growth of sulfate reducing bacteria (SRB) and filamentous sulfur bacteria was monitored on a laboratory scale in activated sludge reactors using acetate and peptone as the artificial wastewater. When the artificial wastewater contained acetate and peptone, filamentous bacteria increased in the sludge and the SVI values increased. There was a good correlation between sulfate reducing activity and sulfur oxidation activity in the produced sludge. The microbial community change of filamentous sulfur bacteria and sulfate reducing bacteria was analyzed using the fluorescent in situ hybridization (FISH) method. The tendency for the growth of filamentous sulfur bacteria Thiothrix eikelboomii following the growth of SRB was observed. The percentage of SRB385- hybridized cells and DNMA657-hybridized cells found in the total cell area increased from 2-3% to 7-10% when the filamentous bulking occurred.

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