Bacterial community analysis of activated sludge: an evaluation of four commonly used DNA extraction methods

The effectiveness of three commercially available direct DNA isolation kits (Mobio, Fast, Qiagen) and one published direct DNA extraction protocol (Bead) for extracting bacterial DNA from different types of activated sludge was investigated and mutually compared. The DNA quantity and purity were determined using real-time PCR targeting the bacterial 16S rDNA gene. Microbial community fingerprints were assessed by automated ribosomal intergenic spacer analysis. The resulting community profiles were analyzed with canonical correspondence analysis. Our results clearly demonstrate that direct DNA extraction methods can significantly influence the DNA quantity, purity, and observed community patterns of microbiota in activated sludge. Fast and Mobio generated high amounts of good quality DNA compared to Bead and Qiagen. Mobio also resulted in the detection of the highest number of species while Fast scored the best in discriminating between the community patterns of different activated sludge types. With respect to the characterization of community profiles, our analyses demonstrated a strong sludge type dependent variability among methods. Taking into account our results, we recommend Fast as the most suitable DNA extraction method for activated sludge samples used for bacterial community studies.status: publishe

Novel magnetically induced membrane vibration (MMV) for fouling control in membrane bioreactors

Conventional submerged membrane bioreactors (MBRs) rely on the coarse bubbles aeration to generate shear at the liquid membrane interface to limit membrane fouling. Unfortunately, it is a very energy consuming method, still often resulting in a rapid decrease of membrane permeability and consequently in higher expenses. In this paper, the feasibility of a novel magnetically induced membrane vibration (MMV) system was studied in a labscale MBR treating synthetic wastewater. The effects on membrane fouling of applied electrical power of different operation strategies, of membrane flux and of the presence of multiple membranes on one vibrating engine on membrane fouling were investigated. The filtration performance was evaluated by determining the filtration resistance profiles and critical flux. The results showed clear advantages of the vibrating system over conventional MBR processes by ensuring higher fluxes at lower fouling rates. Intermittent vibration was found a promising strategy for both efficient fouling control and significant energy saving. The optimised MMV system is presumed to lead to significant energy and cost reduction in up-scaled MBR operations. (C) 2011 Elsevier Ltd. All rights reserved.status: publishe

Evaluation of power ultrasound for disinfection of both Legionella pneumophila and its environmental host Acanthamoeba castellanii

The objectives of this study were to (1) examine the effect of power ultrasound on the viability of both Legionella pneumophila and Acanthamoeba castellanii trophozoites and cysts, (2) investigate if intracellular Legionella replication in trophozoites positively affects bacterial resistance to ultrasound and (3) study if Legionella renders viable but non-culturable (VBNC) due to ultrasound treatments. Using laboratory scale experiments, microorganisms were exposed for various time periods to power ultrasound at a frequency of 36 kHz and an ultrasound power setting of 50 and 100%. Due to a fast destruction, trophozoite hosts were not able to protect intracellular Legionella from eradication by ultrasound, in contrast to cysts. No significant effects of ultrasound on cyst viability could be detected and power settings of 100% for 30 min only made intracellular Legionella concentrations decrease with 1.3 log units. Due to intracellular replication of Legionella in trophozoites, ultrasound no longer affected bacterial viability. Concerning the VBNC state, ultrasound treatments using a power setting of 50% partly induced Legionella (+/- 7%) to transform into VBNC bacteria, in contrast to power settings of 100%. Promising results obtained in this study indicate the possible application of power ultrasound in the control of both Legionella and Acanthamoeba concentrations in anthropogenic water systems. (c) 2009 Elsevier Ltd. All rights reserved.status: publishe

Exploring the potential synergistic effects of chemical disinfectants and UV on the inactivation of free-living bacteria and treatment of biofilms in a pilot-scale system

The main objective of this study is to explore possible synergistic or additive effects of combinations of chemical disinfectants (sodium hypochlorite, peracetic acid, hydrogen peroxide, chlorine dioxide) and UV in their efficacy in inactivating free-living bacteria and removing biofilms. In contrast to most studies, this study examines disinfection of municipal water in a pilot-scale system using a mixed bacterial suspension, which enables a better simulation of the conditions encountered in actual industrial environments. It was shown that the combination of either hypochlorite, hydrogen peroxide, peracetic acid, or chlorine dioxide with UV yielded additive effects on the inactivation of free-living bacteria. Actual synergy was observed for the combination of UV and 5 ppm hydrogen peroxide. Regarding biofilm treatment, additive effects were observed using the combination of hydrogen peroxide and UV. The promising results obtained in this study indicate that the combination of UV and chemical disinfectants can considerably reduce the amount of chemicals required for the effective disinfection and treatment of biofilms.status: publishe

The iimpact of electrochemical disinfection on Escherichia coli and Legionella pneumophila in tap water

In order to reduce the risks of Legionnaires’ disease, caused by the bacterium Legionella pneumophila, disinfection of tap water systems contaminated with this bacterium is a necessity. This study investigates if electrochemical disinfection is able to eliminate such contamination. Hereto, water spiked with bacteria (104 CFU Escherichia coli or L. pneumophila/ml) was passed through an electrolysis cell (direct effect) or bacteria were added to tap water after passage through such disinfection unit (residual effect). The spiked tap water was completely disinfected, during passage through the electrolysis cell, even when only a residual free oxidant concentration of 0.07 mg/l is left (L. pneumophila). The residual effect leads to a complete eradication of cultivable E. coli, if after reaction time at least a free oxidant concentration of 0.08 mg/l is still present. Similar conditions reduce substantially L. pneumophila, but a complete killing is not realised.status: publishe

Spherical Nucleic Acid enhanced FO-SPR DNA melting for detection of mutations in Legionella pneumophila

A home-built Fiber Optic Surface Plasmon Resonance platform (FO-SPR) was applied to directly screen PCR amplified DNA for mutations. The FO-SPR sensor was used for real-time monitoring of DNA duplex melting during high resolution temperature cycling. The signal of the DNA melting was enhanced by means of gold nanoparticle labels. This FO-SPR genetic assay allowed for detection of single-point mutations (SNP) in less than 20 minutes. The concept was demonstrated for the analysis of 9 different serogroups of the bacterium Legionella pneumophila, a common human pathogen responsible for atypical pneumonia. FO-SPR allowed to detect genetic mutations inhibiting PCR, which could lead to amplification bias when molecular diagnostics are applied for L. pneumophila detection. All serogroups were found to display unique melting temperatures, indicating that mutations have accumulated in the target sequence. In a next step, clinical samples of L. pneumophila were analyzed using the FO-SPR sensor. This technology was proven to be reliable for the detection of mutations for those samples that previously displayed ambiguous qPCR quantification results. When these results were benchmarked, FO-SPR results were found to be consistent with Sanger sequencing but not with fluorescence based DNA melting. The presented results convincingly advocate the advantages of FO-SPR as a high resolution and fast genetic screening tool that can compete with the current standard techniques for SNP detection.status: publishe

Spherical Nucleic Acid Enhanced FO-SPR DNA Melting for Detection of Mutations in <i>Legionella pneumophila</i>

A home-built fiber optic surface plasmon resonance platform (FO-SPR) was applied to directly screen PCR amplified DNA for mutations. The FO-SPR sensor was used for real-time monitoring of DNA duplex melting during high resolution temperature cycling. The signal of the DNA melting was enhanced by means of gold nanoparticle labels. This FO-SPR genetic assay allowed for detection of single-point mutations (SNP) in less than 20 min. The concept was demonstrated for the analysis of 9 different serogroups of the bacterium <i>Legionella pneumophila</i>, a common human pathogen responsible for atypical pneumonia. FO-SPR allowed us to detect genetic mutations inhibiting PCR, which could lead to amplification bias when molecular diagnostics are applied for <i>L. pneumophila</i> detection. All serogroups were found to display unique melting temperatures, indicating that mutations have accumulated in the target sequence. In a next step, clinical samples of <i>L. pneumophila</i> were analyzed using the FO-SPR sensor. This technology was proven to be reliable for the detection of mutations for those samples that previously displayed ambiguous qPCR quantification results. When these results were benchmarked, FO-SPR results were found to be consistent with Sanger sequencing but not with fluorescence based DNA melting. The presented results convincingly advocate the advantages of FO-SPR as a high resolution and fast genetic screening tool that can compete with the current standard techniques for SNP detection