A simple optode based method for imaging O<inf>2</inf> distribution and dynamics in tap water biofilms

A ratiometric luminescence intensity imaging approach is presented, which enables spatial O2 measurements in biofilm reactors with transparent planar O2 optodes. Optodes consist of an O2 sensitive luminescent dye immobilized in a 1-10 μm thick polymeric layer on a transparent carrier, e.g. a glass window. The method is based on sequential imaging of the O2 dependent luminescence intensity, which are subsequently normalized with luminescent intensity images recorded under anoxic conditions. We present 2-dimensional O2 distribution images at the base of a tap water biofilm measured with the new ratiometric method and compare the results with O2 distribution images obtained in the same biofilm reactor with luminescence lifetime imaging. Using conventional digital cameras, such simple normalized luminescence intensity imaging can yield images of 2-dimensional O2 distributions with a high signal-to-noise ratio and spatial resolution comparable or even surpassing those obtained with expensive and complex luminescence lifetime imaging systems. The method can be applied to biofilm growth incubators allowing intermittent experimental shifts to anoxic conditions or in systems, in which the O2 concentration is depleted during incubation. © 2011 Elsevier Ltd

Quantitative measurement and visualization of biofilm O <inf>2</inf> consumption rates in membrane filtration systems

There is a strong need for techniques enabling direct assessment of biological activity of biofouling in membrane filtration systems. Here we present a new quantitative and non-destructive method for mapping O 2 dynamics in biofilms during biofouling studies in membrane fouling simulators (MFS). Transparent planar O 2 optodes in combination with a luminescence lifetime imaging system were used to map the two-dimensional distribution of O 2 concentrations and consumption rates inside the MFS. The O 2 distribution was indicative for biofilm development. Biofilm activity was characterized by imaging of O 2 consumption rates, where low and high activity areas could be clearly distinguished. The spatial development of O 2 consumption rates, flow channels and stagnant areas could be determined. This can be used for studies on concentration polarization, i.e. salt accumulation at the membrane surface resulting in increased salt passage and reduced water flux. The new optode-based O 2 imaging technique applied to MFS allows non-destructive and spatially resolved quantitative biological activity measurements (BAM) for on-site biofouling diagnosis and laboratory studies. The following set of complementary tools is now available to study development and control of biofouling in membrane systems: (i) MFS, (ii) sensitive pressure drop measurement, (iii) magnetic resonance imaging, (iv) numerical modelling, and (v) biological activity measurement based on O 2 imaging methodology. © 2011 Elsevier B.V

Online flow cytometry reveals microbial dynamics influenced by concurrent natural and operational events in groundwater used for drinking water treatment

Detailed measurements of physical, chemical and biological dynamics in groundwater are key to understanding the important processes in place and their influence on water quality - particularly when used for drinking water. Measuring temporal bacterial dynamics at high frequency is challenging due to the limitations in automation of sampling and detection of the conventional, cultivation-based microbial methods. In this study, fully automated online flow cytometry was applied in a groundwater system for the first time in order to monitor microbial dynamics in a groundwater extraction well. Measurements of bacterial concentrations every 15 minutes during 14 days revealed both aperiodic and periodic dynamics that could not be detected previously, resulting in total cell concentration (TCC) fluctuations between 120 and 280 cells mu L-1. The aperiodic dynamic was linked to river water contamination following precipitation events, while the (diurnal) periodic dynamic was attributed to changes in hydrological conditions as a consequence of intermittent groundwater extraction. Based on the high number of measurements, the two patterns could be disentangled and quantified separately. This study i) increases the understanding of system performance, ii) helps to optimize monitoring strategies, and iii) opens the possibility for more sophisticated (quantitative) microbial risk assessment of drinking water treatment systems

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Controversy exists as to whether neoadjuvant chemotherapy improves survival in patients with invasive bladder cancer, despite randomised controlled trials of more than 3000 patients. We undertook a systematic review and meta-analysis to assess the effect of such treatment on survival in patients with this disease

Absolute quantification of microbial taxon abundances

High-throughput amplicon sequencing has become a well-established approach for microbial community profiling. Correlating shifts in the relative abundances of bacterial taxa with environmental gradients is the goal of many microbiome surveys. As the abundances generated by this technology are semi-quantitative by definition, the observed dynamics may not accurately reflect those of the actual taxon densities. We combined the sequencing approach (16S rRNA gene) with robust single-cell enumeration technologies (flow cytometry) to quantify the absolute taxon abundances. A detailed longitudinal analysis of the absolute abundances resulted in distinct abundance profiles that were less ambiguous and expressed in units that can be directly compared across studies. We further provide evidence that the enrichment of taxa (increase in relative abundance) does not necessarily relate to the outgrowth of taxa (increase in absolute abundance). Our results highlight that both relative and absolute abundances should be considered for a comprehensive biological interpretation of microbiome surveys