Bacterial water quality and network hydraulic characteristics: a field study of a small, looped water distribution system using culture-independent molecular methods.

AIMS: To determine the spatial and temporal variability in the abundance, structure and composition of planktonic bacterial assemblages sampled from a small, looped water distribution system and to interpret results with respect to hydraulic conditions. METHODS AND RESULTS: Water samples were collected from five sampling points, twice a day at 06:00 h and 09:00 h on a Monday (following low weekend demand) and a Wednesday (higher midweek demand). All samples were fully compliant with current regulated parameter standards. This study did not show obvious changes in bacterial abundance (DAPI count) or community structure Denaturing gradient gel electrophoresis analysis with respect to sample site and hence to water age; however, the study did show temporal variability with respect to both sampling day and sample times. CONCLUSIONS: Data suggests that variations in the bacterial assemblages may be associated with the local system hydraulics: the bacterial composition and numbers, over short durations, are governed by the interaction of the bulk water and the biofilm influenced by the hydraulic conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: This study demonstrates general stability in bacterial abundance, community structure and composition within the system studied. Trends and patterns supporting the transfer of idealized understanding to the real world were evident. Ultimately, such work will help to safeguard potable water quality, fundamental to public health.

Bacterial quorum sensing and cell surface electrokinetic properties.

The hypothesis tested in this paper is that quorum sensing influences the microbial surface electrokinetic properties. Escherichia coli MG1655 and MG1655 LuxS- mutant (lacking quorum-sensing gene for Autoinducer synthase AI-2) were used for this study. AI-2 production (or lack of) in both strains was analyzed using the Vibrio harveyi bioassay. The levels of extracellular AI-2 with and without glucose in the growth medium were consistent with previously published work. The surface electrokinetic properties were determined for each strain of E. coli MG1655 by measuring the electrophoretic mobility using a phase amplitude light-scattering (PALS) Zeta potential analyser. The findings show that the surface charge of the cells is dependent upon the stage in the growth phase as well as the ability to participate in quorum sensing. In addition, significant differences in the electrophoretic mobility were observed between both strains of E. coli. These findings suggest that quorum sensing plays a significant role in the surface chemistry of bacteria during their growth.

Role of nonadsorbing polymers in bacterial aggregation.

Bacteria exhibit properties similar to those of nonbiological colloids and can display pairwise attractions when in close proximity. This interaction is governed by the surface chemistry of the cells. We seek to understand bacterial aggregation at the cellular level using Escherichia coli (E. coli) AB1157. Aggregation studies were carried out using 0.5 to 2.5 wt% E. coli AB1157 harvested in different growth phases with varying concentrations of a nonadsorbing polymer, sodium polystyrene sulfonate (SPS). The electrophoretic mobility of E. coli AB1157 in different growth phases was determined using phase-amplitude light scattering. E. coli AB1157 was found to be negatively charged, and the cell surface properties changed in different growth phases. The electrokinetic results correlated well with the different concentrations of nonadsorbing polymer needed to induce depletion aggregation. This shows that a difference in aggregation properties is due to changes in the bacteria electrokinetic properties during their growth.

Investigating the effect of storm events on the particle size distribution in a combined sewer simulator.

The changes in particle size of sewer sediment particles rapidly eroded from a previously deposited sediment bed are described, using a rotating annular flume as a laboratory scale sewer simulator. This is the first time that particle size distributions of eroded sewer sediments from a previously deposited sediment bed have been monitored in such a controlled experimental environment. Sediments from Loenen, The Netherlands and Dundee, UK were used to form deposits in the base of the annular flume (WL Delft Netherlands) with varying conditions for consolidation in order to investigate the effect of changing consolidation time, temperature and sediment type on the amount and size of particles eroded from a bed under conditions of increasing shear. The median size of the eroded particles did not change significantly with temperature, although the eroded suspended solids concentration was greater for the higher temperature under the same shear stresses, indicating a weaker bed deposit. An increase in consolidation time caused an increase in median size of eroded solids at higher bed shear stresses, and this was accompanied by higher suspended solids concentrations. As the shear stress increased, the solids eroded from the bed developed under a longer consolidation time (56 hours) tended towards a broad unimodal distribution, whilst the size distribution of solids eroded from beds developed under shorter consolidation times (18 or 42 hours) retained a bi- or tri-modal distribution. Using different types of sediment in the flume had a marked effect on the size of particles eroded.

Modelling the activated sludge flocculation process combining laser light diffraction particle sizing and population balance modelling (PBM).

A technique based on laser light diffraction is shown to be successful in collecting on-line experimental data. Time series of floc size distributions (FSD) under different shear rates (G) and calcium additions were collected. The steady state mass mean diameter decreased with increasing shear rate G and increased when calcium additions exceeded 8 mg/l. A so-called population balance model (PBM) was used to describe the experimental data. This kind of model describes both aggregation and breakage through birth and death terms. A discretised PBM was used since analytical solutions of the integro-partial differential equations are non-existing. Despite the complexity of the model, only 2 parameters need to be estimated; the aggregation rate and the breakage rate. The model seems, however, to lack flexibility. Also, the description of the floc size distribution (FSD) in time is not accurate.

Activated sludge flocculation: direct determination of the effect of calcium ions.

The effect of calcium on activated sludge flocculation dynamics is investigated using a unique experimental technique. The technique allows on-line analysis of the size of activated sludge flocs during flocculation and provides valuable insight into the mechanisms of flocculation. Activated sludge samples were firstly sonicated for 3 minutes at 50 W and then stirred at 100 rpm. The floc size was subsequently measured on-line using a Malvern Mastersizer/E. For concentrations of calcium less than 4 meq/L no significant increase in final floc size was observed even though an increase in the initial rate of change of floc size could be seen. Addition of calcium greater than 4 meq/L resulted in a dramatic increase in floc size. Results from this investigation support the theory that cations are involved in flocculation through cationic bridging, and will be used in ongoing investigations to model the flocculation process.

Reverse transcriptase PCR amplification of EWS/FLI-1 fusion transcripts as a diagnostic test for peripheral primitive neuroectodermal tumors of childhood.

The peripheral primitive neuroectodermal tumors (pPNETs) of childhood, including Ewing's sarcoma, peripheral neuroepithelioma, and Askin's tumor, often present significant diagnostic challenges for the anatomic pathologist. One consistent feature of these tumors is the presence of the t(11;22)(q24;q12) in tumor cells, and this translocation has been useful as a marker for this group of tumors. The recent cloning of the t(11;22) breakpoint has revealed the fusion of the human FLI-1 gene on chromosome 11q24 with a gene of unknown function called EWS on 22q12, and fusion transcripts have been detected. These findings have raised the possibility of using molecular genetic analysis as a tool to diagnose pPNETs. To this end, we have tested pPNETs for the presence of EWS/FLI-1 fusion transcripts by reverse transcriptase-polymerase chain reaction (RT-PCR) using EWS and FLI-1 specific primers. Eight (80%) of 10 pPNET cell lines were positive for amplified products using this technique. These results were confirmed by Southern analysis, which revealed rearrangements of EWS using genomic EWS probes in all eight positive cell lines. We then tested 20 primary pPNET tumors, and identified fusion transcripts by RT-PCR in 18 (90%) of these cases. Cloning and sequencing of PCR products confirmed the presence of EWS and FLI-1 sequences in these products. Furthermore, fusion transcripts were not detected by this technique in a series of non-pPNET pediatric solid tumors. Detection of EWS/FLI-1 fusion transcripts by RT-PCR therefore provides a novel adjunctive tool in the diagnosis of pPNETs.

The medical and economic impact of closed cervical spine dislocations.

The current study examined the medical and economic impact of various treatment modalities of unstable cervical spine dislocations. Thirty-five patients with dislocations of the cervical spine were reviewed retrospectively from two large university hospitals. There were 23 unilateral and 12 bilateral facet dislocations. Fourteen patients were treated nonoperatively and 21 patients were treated with early surgery. Nine of the 14 patients treated nonoperatively redisplaced their dislocations while in an orthosis and required late surgery. Analysis of variance shows that it is the amount of injury (unilateral versus bilateral) that results in statistically significant difference in the injury severity score, Glasgow coma scale, trauma motor index, and the number of complications. Of the treatment groups (operative versus nonoperative), the operative group showed a statistically significant difference in the length of hospital stay. Also noted in the operative group was less traction time, earlier erect physical therapy, an average financial savings of $18,407.00, and equal morbidity or mortality to the nonoperative treatment group. Based on this limited experience, it would appear that "diagnosis related groups" are not capable of supporting traumatic spinal care and, if adopted by other states and third party payers, the concept of trauma and/or spinal cord injury centers may be in jeopardy.