Revealing microbial community structures in large- and small-scale activated sludge systems by barcoded pyrosequencing of 16S rRNA gene

2012 ◽  
Vol 66 (10) ◽  
pp. 2155-2161 ◽  
Author(s):  
Purnika Damindi Ranasinghe ◽  
Hiroyasu Satoh ◽  
Mamoru Oshiki ◽  
Kenshiro Oshima ◽  
Wataru Suda ◽  
...  

The diversity of bacterial groups in activated sludge from large- and small-scale wastewater treatment plants was explored by barcoded pyrosequencing of 16S rRNA gene. Activated sludge samples (three small and 17 large scale) were collected from 12 wastewater treatment plants to clarify precise taxonomy and relative abundances. DNA was extracted, and amplified by 4 base barcoded 27f/519r primer set. The 454 Titanium (Roche) pyrosequences were obtained and analyses performed by Quantitative Insight Into Microbial Ecology (QIIME) with around 100,000 reads. Sequence statistics were computed, while constructing a phylogenetic tree and heatmap. Computed results explained total microbial diversity at phylum and class level and resolution was further extended to Operational Taxonomic Unit (OTU) based taxonomic assignment for investigating community distribution based on individual sample. Composition of sequence reads were compared and microbial community structures for large- and small-scale treatment plants were identified as major phyla (Proteobacteria and Bacteroidetes) and classes (Betaproteobacteria and Bacteroidetes). Also, family level breakdowns were explained and differences in family Nitrospiraceae and phylum Actinobacteria found at their species level were also illustrated. Thus, the pyrosequencing method provides high resolution insight into microbial community structures in activated sludge that might have been unnoticed with conventional approaches.

2020 ◽  
Vol 148 ◽  
pp. 01002
Author(s):  
Herto Dwi Ariesyady ◽  
Mentari Rizki Mayanda ◽  
Tsukasa Ito

Activated sludge process is one of the wastewater treatment method that is applied for many wastewater types including painting process wastewater of automotive industry. This wastewater is well-known to have high heavy metals concentration which could deteriorate water environment if appropriate performance of the wastewater treatment could not be achieved. In this study, we monitored microbial community diversity in a Painting Biological Treatment (PBT) system. We applied a combination of cultivation and genotypic biological methods based on 16S rRNA gene sequence analysis to identify the diversity of active microbial community. The results showed that active microbes that could grow in this activated sludge system were dominated by Gram-negative bacteria. Based on 16S rRNA gene sequencing analysis, it was revealed that their microbial diversity has close association with Bacterium strain E286, Isosphaera pallida, Lycinibacillus fusiformis, Microbacterium sp., Orchobactrum sp., Pseudomonas guariconensis, Pseudomonas sp. strain MR84, Pseudomonas sp. MC 54, Serpens sp., Stenotrophomonas acidaminiphila, and Xylella fastidiosa with similarity of 86 – 99%. This findings reflects that microbial community in a Painting Biological Treatment (PBT) system using activated sludge process could adapt with xenobiotics in the wastewater and has a wide range of diversity indicating a complex metabolism mechanism in the treatment process.


2007 ◽  
Vol 73 (9) ◽  
pp. 2860-2870 ◽  
Author(s):  
Jonathan P. Allen ◽  
Estella A. Atekwana ◽  
Eliot A. Atekwana ◽  
Joseph W. Duris ◽  
D. Dale Werkema ◽  
...  

ABSTRACT The interdependence between geoelectrical signatures at underground petroleum plumes and the structures of subsurface microbial communities was investigated. For sediments contaminated with light non-aqueous-phase liquids, anomalous high conductivity values have been observed. Vertical changes in the geoelectrical properties of the sediments were concomitant with significant changes in the microbial community structures as determined by the construction and evaluation of 16S rRNA gene libraries. DNA sequencing of clones from four 16S rRNA gene libraries from different depths of a contaminated field site and two libraries from an uncontaminated background site revealed spatial heterogeneity in the microbial community structures. Correspondence analysis showed that the presence of distinct microbial populations, including the various hydrocarbon-degrading, syntrophic, sulfate-reducing, and dissimilatory-iron-reducing populations, was a contributing factor to the elevated geoelectrical measurements. Thus, through their growth and metabolic activities, microbial populations that have adapted to the use of petroleum as a carbon source can strongly influence their geophysical surroundings. Since changes in the geophysical properties of contaminated sediments parallel changes in the microbial community compositions, it is suggested that geoelectrical measurements can be a cost-efficient tool to guide microbiological sampling for microbial ecology studies during the monitoring of natural or engineered bioremediation processes.


2000 ◽  
Vol 42 (3-4) ◽  
pp. 17-22 ◽  
Author(s):  
M. Onuki ◽  
H. Satoh ◽  
T. Mino ◽  
T. Matsuo

In the last decade, molecular biology has made significant progress, and innovative molecular methods have become available to analyze microbial community structures. Among them, we applied the FISH (Fluorescent in situ Hybridization) method to analyze activated sludge in wastewater treatment plants (WWTPs). As a result, domain- or division-level community structures in activated sludge were determined successfully without cultivation. We also applied the PCR (polymerase chain reaction) -DGGE (Denaturing Gradient Gel Electrophoresis) method for laboratory nitrifying sludge in order to investigate more detailed microbial community structure. By this method, genus- or species-level community structures were characterized well. This method was also found to be powerful for monitoring the change of microbial community structures. For example, the behavior of Nitrosomonas group was successfully detected in the reactor with nitrification by the PCR-DGGE method.


2021 ◽  
Author(s):  
Morten Simonsen Dueholm ◽  
Marta Nierychlo ◽  
Kasper Skytte Andersen ◽  
Vibeke Rudkjoebing Joergensen ◽  
Simon Knutsson ◽  
...  

Biological wastewater treatment and an increased focus on resource recovery is fundamental for environmental protection, human health, and sustainable development. Microbial communities are responsible for these processes, but our knowledge of their diversity and function is still poor, partly due to the lack of good reference databases and comprehensive global studies. Here, we sequenced more than 5 million high-quality, full-length 16S rRNA gene sequences from 740 wastewater treatment plants (WWTPs) across the world and used the sequences to construct MiDAS 4, a full-length amplicon sequence variant resolved 16S rRNA gene reference database with a comprehensive taxonomy from the domain to species-level for all references. Using a study-independent amplicon dataset from the Global Water Microbiome Consortium project (269 WWTPs), we showed that the MiDAS 4 database provides much better coverage for bacteria in WWTPs worldwide compared to commonly applied universal references databases, and greatly improved the rate of genus and species-level classification. Hence, MiDAS 4 provides a unifying taxonomy for the majority of prokaryotic diversity in WWTPs globally, which can be used for linking microbial identities with their functions across studies. Taking advantage of MiDAS 4, we carried out an amplicon-based, global-scale microbial community profiling of activated sludge plants using two common sets of primers targeting the V1-V3 and V4 region of the 16S rRNA gene. We found that the V1-V3 primers were generally best suited for this ecosystem, and revealed how environmental conditions and biogeography shape the activated sludge microbiota. We also identified process-critical taxa (core and conditionally rare or abundant taxa), encompassing 966 genera and 1530 species. These represented approximately 80% and 50% of the accumulated read abundance, respectively, and represent targets for further investigations. Finally, we showed that for well-studied functional guilds, such as nitrifiers or polyphosphate accumulating organisms, the same genera were prevalent worldwide, with only a few abundant species in each genus.


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