Bacterial diversity dynamics in a long-term petroleum-contaminated soil.

Bacterial diversity dynamics were investigated in the soil samples in different distances and depths from/at a long-term petroleum-contaminated site. Microbial activity in the soil samples showed ATP values closely correlated with organic matter content (OC) and total petroleum hydrocarbon (TPH). Bacterial community diversity (H) and evenness (J) using PCR-DGGE (polymerase chain reaction-denaturing gradient gel electrophoresis) and PCR-T-RFLP (terminal restriction fragment length polymorphism) results showed positive correlation with concentration of TPH or OC, but tmoA (toluene monooxygenase gene)-based bacterial H and J using a PCR-T-RFLP result did not. No significant difference of H and J values in the bacterial and the tmoA communities was observed. The bacterial community structure characterized by PCR-DGGE and PCR-T-RFLP techniques showed similarity according to soil sampling distance rather than soil sampling depth. Canonical correspondence analysis demonstrated that OC including TPH had the most significant effect on the bacterial community diversity at the long-term petroleum-contaminated site.

Comparison of sludge characteristics and PCR-DGGE based microbial diversity of nanofiltration and microfiltration membrane bioreactors.

The objective of this study was to compare the sludge characteristics and microbial community diversity between the submerged nanofiltration membrane bioreactor (NF MBR) and microfiltration membrane bioreactor (MF MBR) treating the same municipal wastewater. The influence of a higher concentration of organic matter and salt was investigated. The results of water qualities showed that the dissolved organic carbon (DOC), total phosphorus (T-P) and salt concentrations of the supernatant in the NF MBR were three, four and two times as high as those in the MF MBR, respectively. The specific oxygen uptake rate of the NF MBR (2.9+/-0.4 mg O(2)g(-1)MLSSh(-1)) was lower than that of the MF MBR (4.3+/-1.1 mg O(2)g(-1)MLSS h(-1)). Result of extractable extracellular polymeric substances showed that the NF MBR sludge had more protein and less polysaccharide compared to the MF MBR sludge, whereas specific amount of total organic carbon were the same in both MBRs. The median floc diameters of the NF MBR and the MF MBR were 72+/-12 microm and 59+/-12 microm, respectively, which could be attributable to the different polysaccharide concentrations between both MBR mixed liquor. A higher concentration of materials (DOC, T-P and salt) in the bioreactor, determined by the high rejection rate of the NF membrane, did not significantly affect the microbial diversity under similar operation conditions.

Population dynamics of phage-host system of Microlunatus phosphovorus indigenous in activated sludge.

Monitoring of the phage-host system of Microlunatus phosphovorus indigenous in activated sludge was attempted. A laboratory-scale activated sludge process was operated for 5 weeks with synthetic wastewater. The phage-host system population in the process was monitored by plaque assay and FISH methods at every 3 days. During the process operation, the phage-host system populations were more or less steady, except for 1 week in the middle of the operation. In that period, initially M. phosphovorus decreased significantly and its lytic bacteriophages increased, and then M. phosphovorus increased back to its original level while its lytic bacteriophages decreased. This observation suggests that lytic bacteriophages should be considered as one of the biological factors affecting the bacterial population dynamics in activated sludge processes.

Gene Expression Correlates with Process Rates Quantified for Sulfate- and Fe(III)-Reducing Bacteria in U(VI)-Contaminated Sediments.

Though iron- and sulfate-reducing bacteria are well known for mediating uranium(VI) reduction in contaminated subsurface environments, quantifying the in situ activity of the microbial groups responsible remains a challenge. The objective of this study was to demonstrate the use of quantitative molecular tools that target mRNA transcripts of key genes related to Fe(III) and sulfate reduction pathways in order to monitor these processes during in situ U(VI) remediation in the subsurface. Expression of the Geobacteraceae-specific citrate synthase gene (gltA) and the dissimilatory (bi)sulfite reductase gene (dsrA), were correlated with the activity of iron- or sulfate-reducing microorganisms, respectively, under stimulated bioremediation conditions in microcosms of sediments sampled from the U.S. Department of Energy's Oak Ridge Integrated Field Research Challenge (OR-IFRC) site at Oak Ridge, TN, USA. In addition, Geobacteraceae-specific gltA and dsrA transcript levels were determined in parallel with the predominant electron acceptors present in moderately and highly contaminated subsurface sediments from the OR-IFRC. Phylogenetic analysis of the cDNA generated from dsrA mRNA, sulfate-reducing bacteria-specific 16S rRNA, and gltA mRNA identified activity of specific microbial groups. Active sulfate reducers were members of the Desulfovibrio, Desulfobacterium, and Desulfotomaculum genera. Members of the subsurface Geobacter clade, closely related to uranium-reducing Geobacter uraniireducens and Geobacter daltonii, were the metabolically active iron-reducers in biostimulated microcosms and in situ core samples. Direct correlation of transcripts and process rates demonstrated evidence of competition between the functional guilds in subsurface sediments. We further showed that active populations of Fe(III)-reducing bacteria and sulfate-reducing bacteria are present in OR-IFRC sediments and are good potential targets for in situ bioremediation.

Microbial treatment of high-strength perchlorate wastewater.

To treat wastewater containing high concentrations of perchlorate, a perchlorate reducing-bacterial consortium was obtained by enrichment culture grown on high-strength perchlorate (1200 mg L(-1)) feed medium, and was characterized in a sequence batch reactor (SBR) over a long-time operation. The consortium removed perchlorate in the SBR with high reduction rates (35-90 mg L(-1)h(-1)) and stable removal efficiency over 200-day operations. The maximum specific perchlorate reduction rate (qmax), half saturation constant (Ks), and optimal pH range were 0.67 mg-perchlorate mg-dry cell weight(-1) h(-1), 193.8 mg-perchlorate L(-1), and pH 7-9, respectively. The perchlorate reduction yield was 0.48 mol-perchloratemol-acetate(-1). A clone library prepared using the amplicons of cld gene encoding chlorate dismutase showed that the dominant (per)chlorate reducing bacteria in the consortium were Dechlorosoma sp. (53%), Ideonella sp. (28%), and Dechloromonas sp. (19%).

Earthworm cast as a promising filter bed material and its methanotrophic contribution to methane removal.

The use of biocovers is a promising strategy toward mitigating CH(4) emission from smaller and/or older landfills. In this study, a filter bed material consisting of a mixture of earthworm cast and rice paddy soil in a biocover was evaluated. Although the CH(4) oxidation rate of the enriched paddy soil was 4.9 microg g-dry soil(-1) h(-1), it was enhanced to 25.1 microg g-dry soil(-1) h(-1) by adding an earthworm cast with a 3:7 ratio of earthworm cast:soil (wet weight). CO(2) was found as the final oxidation product of CH(4), and the mole ratio of CO(2) production to CH(4) consumption was 0.27. At a moisture content range of 15-40% and a temperature range of 20-40 degrees C, the CH(4) oxidation rates of the enriched mixture were more than 57% of the maximum rate obtained at 25% moisture content and 25 degrees C. By denaturing gradient gel electrophoresis analysis employing primers for the universal bacterial 16S rRNA gene, and terminal-restriction fragment length polymorphism analysis using primers for the pmoA gene, the bacterial and methanotrophic communities in the enriched mixture were mainly originate from paddy soil and earthworm cast, respectively. Both type I (mainly Methylocaldum) and type II methanotrophs (mainly Methylocystis) played important roles in CH(4) oxidation in the enriched mixture.

Abundance, diversity, and dynamics of viruses on microorganisms in activated sludge processes.

We examined the abundance of viruses on microorganisms in activated sludge and the dynamics of their community structure. Direct counting with epifluorescence microscopy and pulsed-field gel electrophoresis (PFGE) were applied to 20 samples from 14 full-scale wastewater treatment plants (wwtps) treating municipal, industrial, or animal wastewater. Furthermore, to observe the dynamics of viral community structure over time, a laboratory-scale sequencing batch reactor was operated for 58 days. The concentrations of virus particles in the wwtps, as quantified by epifluorescence microscopy, ranged from 4.2 x 10(7) to 3.0 x 10(9) mL-1. PFGE, improved by the introduction of a higher concentration of Tris-EDTA buffer in the DNA extraction step, was successfully used to profile DNA viruses in the activated sludge. Most of the samples from different wwtps commonly had bands in the 40-70 kb range. In the monitoring of viral DNA size distribution in the laboratory-scale reactor, some bands were observed stably throughout the experimental period, some emerged during the operation, and others disappeared. Rapid emergence and disappearance of two intense bands within 6 days was observed. Our data suggest that viruses--especially those associated with microorganisms--are abundant and show dynamic behavior in activated sludge.