Microbial community analysis of a full-scale membrane bioreactor treating industrial wastewater

2008 ◽  
Vol 58 (8) ◽  
pp. 1589-1594 ◽  
Author(s):  
D. Naidoo ◽  
N. Ramdhani ◽  
F. Bux
Keyword(s):  
In Situ Hybridization ◽  
Microbial Community ◽  
Manufacturing Industry ◽  
Community Analysis ◽  
Microbial Community Analysis ◽  
Gram Positive ◽  
Gram Positive Bacteria ◽  
Positive Hybridization ◽  
Gram Negative Organisms

A Kubota™ submerged membrane bio-reactor was applied to treat wastewater from a sugar manufacturing industry. To achieve optimal results, fundamental and extended understanding of the microbiology is important. Fluorescence in situ hybridization was used to evaluate the microbial community present. The majority of cells visualized in the sludge flocs by staining with the DNA fluorochrome DAPI, hybridized strongly with a bacterial probe. Probes specific for the alpha-, beta-, and gamma-subclasses of proteobacteria and high G + C Gram positive bacteria were used to characterize the community structures by in situ hybridization. Sampling was carried out over 12 weeks and samples were fixed with 4% paraformaldehyde for gram positive organisms and ice cold ethanol for gram negative organisms. The activated sludge population usually constitutes about 80 to 90% of proteobacteria. However, in this study it was found that a relatively small amount of proteobacteria was present within the system. No positive hybridization signal was observed with any of the applied eubacterial family- level probes.

scholarly journals Community Analysis of Biofilters Using Fluorescence In Situ Hybridization Including a New Probe for theXanthomonas Branch of the ClassProteobacteria

1999 ◽  
Vol 65 (8) ◽  
pp. 3547-3554 ◽  
Author(s):  
Udo Friedrich ◽  
Michèle M. Naismith ◽  
Karlheinz Altendorf ◽  
André Lipski
Keyword(s):  
In Situ Hybridization ◽  
Community Analysis ◽  
Specific Probe ◽  
Efficient Tool ◽  
Gram Positive ◽  
Detection Rates ◽  
Gram Positive Bacteria ◽  
Practical Tool ◽  
Direct Counts

ABSTRACT Domain-, class-, and subclass-specific rRNA-targeted probes were applied to investigate the microbial communities of three industrial and three laboratory-scale biofilters. The set of probes also included a new probe (named XAN818) specific for the Xanthomonasbranch of the class Proteobacteria; this probe is described in this study. The members of the Xanthomonas branch do not hybridize with previously developed rRNA-targeted oligonucleotide probes for the α-, β-, and γ-Proteobacteria. Bacteria of the Xanthomonas branch accounted for up to 4.5% of total direct counts obtained with 4′,6-diamidino-2-phenylindole. In biofilter samples, the relative abundance of these bacteria was similar to that of the γ-Proteobacteria. Actinobacteria(gram-positive bacteria with a high G+C DNA content) and α-Proteobacteria were the most dominant groups. Detection rates obtained with probe EUB338 varied between about 40 and 70%. For samples with high contents of gram-positive bacteria, these percentages were substantially improved when the calculations were corrected for the reduced permeability of gram-positive bacteria when formaldehyde was used as a fixative. The set of applied bacterial class- and subclass-specific probes yielded, on average, 58.5% (± a standard deviation of 23.0%) of the corrected eubacterial detection rates, thus indicating the necessity of additional probes for studies of biofilter communities. The Xanthomonas-specific probe presented here may serve as an efficient tool for identifying potential phytopathogens. In situ hybridization proved to be a practical tool for microbiological studies of biofiltration systems.

The microbial community analysis of a 5-stage BNR process with step feed system

2003 ◽  
Vol 48 (8) ◽  
pp. 135-141 ◽  
Author(s):  
H.W. Lee ◽  
S.Y. Lee ◽  
J.O. Lee ◽  
H.G. Kim ◽  
J.B. Park ◽  
...  
Keyword(s):  
16S Rdna ◽  
Community Analysis ◽  
Cell Number ◽  
Total Cell ◽  
Microbial Community Analysis ◽  
Feed System ◽  
Gram Positive ◽  
Gram Positive Bacteria ◽  
Cell Counts

The microbial communities of 5-stage BNR activated sludge samples were analyzed using fluorescence in-situ hybridization (FISH) and 16S rDNA characterization. The total cell numbers of each reactor were from 2.36 × 109 cells/ml to 2.83 × 109 cells/ml. From 56.5% to 62.0% of total DAPI cell counts were hybridized to the most bacterial specific probe EUB 338. Among them, b-proteobacteria were most dominant in each tank. The number of phosphate accumulating organisms (PAOs) was almost 50% of the total cell number in anoxic-1 tank, and these results indicate that this process has a high content of denitrifying phosphorus accumulating organisms (dPAOs). In contrast with FISH, 16S rDNA analysis showed that dominant groups were the Cytophaga-Flavobacterium group and high G+C% gram-positive bacteria, which were determined as PAOs in anoxic-1 tank. The beta subclass Proteobacteria did not accumulate a large amount of polyphosphate. The overall results indicate that high G+C% gram-positive bacteria and the Cytophaga-Flavobacterium group might play a key role as dPAOs in this process.

Lysozyme Treatment Makes 16S rRNA Amplicon Sequencing Results Less Biased

2021 ◽  
Author(s):  
Yuting Jiao ◽  
Zijie Gao ◽  
Shiyu Gui ◽  
Lu Ren ◽  
Yongyue Lu ◽  
...  
Keyword(s):  
Structure Analysis ◽  
Community Analysis ◽  
Amplicon Sequencing ◽  
Microbial Community Analysis ◽  
Effective Agent ◽  
Gram Positive ◽  
Gram Positive Bacteria ◽  
16S Rrna Amplicon Sequencing ◽  
Structure Result ◽  
Total Bacteria

Abstract Background Amplicon sequencing is widely applied in gut bacteria structure analysis. However, the proportion of Gram-positive bacteria may greatly affect the results of microbial community analysis. Lysozyme is an effective agent to extract DNA of Gram-positive bacteria. In this study, we assessed the influence of lysozyme treatment on results of Bactrocere dorsalis rectal bacteria structure. Result The results indicated that the total bacteria content can be significantly increased in lysozyme treated samples. Moreover, rectal bacteria diversity was significantly higher in lysozyme treated samples. A detail analysis revealed that abundance of Gram-positive bacteria significantly increased in samples treated with lysozyme. Conclusion This study indicates that lysozyme treatment before DNA extraction is an effective way to reduce bias in bacteria structure analysis, especially for samples with high proportion of Gram-positive bacteria.

Estimation of the State of the Bacterial Cell Wall by Fluorescent In Situ Hybridization

1998 ◽  
Vol 64 (8) ◽  
pp. 3059-3062 ◽  
Author(s):  
Elena Bidnenko ◽  
Carine Mercier ◽  
Josselyne Tremblay ◽  
Patrick Tailliez ◽  
Saulius Kulakauskas
Keyword(s):  
Cell Wall ◽  
In Situ Hybridization ◽  
Cell Walls ◽  
Fluorescent In Situ Hybridization ◽  
Cell Level ◽  
Gram Positive ◽  
Gram Positive Bacteria ◽  
A Cell ◽  
Identification Of Bacteria

ABSTRACT Fluorescent in situ hybridization (FISH) is now a widely used method for identification of bacteria at the single-cell level. With gram-positive bacteria, the thick peptidoglycan layer of a cell wall presents a barrier for entry of horseradish peroxidase (HRP)-labeled probes. Therefore, such probes do not give any signal in FISH unless cells are first treated with enzymes which hydrolyze the peptidoglycan. We explored this feature of FISH to detect cells which have undergone permeabilization due to expression of autolytic enzymes. Our results indicate that FISH performed with HRP-labeled probes provides a sensitive method to estimate the states of cell walls of individual gram-positive bacteria.

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