Effect of flow regimes on the presence of Legionella within the biofilm of a model plumbing system

AIMS: Stagnation is widely believed to predispose water systems to colonization by Legionella. A model plumbing system was constructed to determine the effect of flow regimes on the presence of Legionella within microbial biofilms. METHODS AND RESULTS: The plumbing model contained three parallel pipes where turbulent, laminar and stagnant flow regimes were established. Four sets of experiments were carried out with Reynolds number from 10,000 to 40,000 and from 355 to 2,000 in turbulent and laminar pipes, respectively. Legionella counts recovered from biofilm and planktonic water samples of the three sampling pipes were compared with to determine the effect of flow regime on the presence of Legionella. Significantly higher colony counts of Legionella were recovered from the biofilm of the pipe with turbulent flow compared with the pipe with laminar flow. The lowest counts were in the pipe with stagnant flow. CONCLUSIONS: We were unable to demonstrate that stagnant conditions promoted Legionella colonization. SIGNIFICANCE AND IMPACT OF THE STUDY: Plumbing modifications to remove areas of stagnation including deadlegs are widely recommended, but these modifications are tedious and expensive to perform. Controlled studies in large buildings are needed to validate this unproved hypothesis.     

THE EFFECT OF THIOSULPHATE AND OF PHOSPHATE ON THE BACTERICIDAL ACTION OF COPPER AND ZINC IN SAMPLES OF WATER

SUMMARY: Because of the increased use of copper and galvanized pipes, samples of water for bacteriological examination not infrequently contain minute traces of copper or zinc. These metals have a fairly rapid bactericidal action in the sampling bottle. The bactericidal action of copper can be prevented by the thiosulphate used for neutralizing chlorine; 100 mg/l is a useful quantity for general use in all sampling bottles. The bactericidal action from zinc can be prevented by the use of a small quantity of sodium phosphate in the sampling bottle. When special sampling points are installed, copper or galvanized pipes should be avoided.     

A family of pneumatically-operated thin layer samplers for replicate sampling of heterogeneous water columns

A series of inexpensive, pneumatic thin layer water samplers is described. They can be operated from small boats, and permit sampling at 2.5 cm intervals with little or no disturbance of stratified systems such as oxyclines or redoxclines of meromictic lakes, or microstratification of flagellates in sheltered epilimnia. Some models permit replicate sampling at closely-spaced intervals in a two-dimensional array. Their performance abilities are illustrated with examples of microstratification.     

A new coupon design for simultaneous analysis of in situ microbial biofilm formation and community structure in drinking water distribution systems

This study presents a new coupon sampling device that can be inserted directly into the pipes within water distribution systems (WDS), maintaining representative near wall pipe flow conditions and enabling simultaneous microscopy and DNA-based analysis of biofilms formed in situ. To evaluate this sampling device, fluorescent in situ hybridization (FISH) and denaturing gradient gel electrophoresis (DGGE) analyses were used to investigate changes in biofilms on replicate coupons within a non-sterile pilot-scale WDS. FISH analysis demonstrated increases in bacterial biofilm coverage of the coupon surface over time, while the DGGE analysis showed the development of increasingly complex biofilm communities, with time-specific clustering of these communities. This coupon design offers improvements over existing biofilm sampling devices in that it enables simultaneous quantitative and qualitative compositional characterization of biofilm assemblages formed within a WDS, while importantly maintaining fully representative near wall pipe flow conditions. Hence, it provides a practical approach that can be used to capture the interactions between biofilm formation and changing abiotic conditions, boundary shear stress, and turbulent driven exchange within WDS.     

Temporal Variations in the Abundance and Composition of Biofilm Communities Colonizing Drinking Water Distribution Pipes

Pipes that transport drinking water through municipal drinking water distribution systems (DWDS) are challenging habitats for microorganisms. Distribution networks are dark, oligotrophic and contain disinfectants; yet microbes frequently form biofilms attached to interior surfaces of DWDS pipes. Relatively little is known about the species composition and ecology of these biofilms due to challenges associated with sample acquisition from actual DWDS. We report the analysis of biofilms from five pipe samples collected from the same region of a DWDS in Florida, USA, over an 18 month period between February 2011 and August 2012. The bacterial abundance and composition of biofilm communities within the pipes were analyzed by heterotrophic plate counts and tag pyrosequencing of 16S rRNA genes, respectively. Bacterial numbers varied significantly based on sampling date and were positively correlated with water temperature and the concentration of nitrate. However, there was no significant relationship between the concentration of disinfectant in the drinking water (monochloramine) and the abundance of bacteria within the biofilms. Pyrosequencing analysis identified a total of 677 operational taxonomic units (OTUs) (3% distance) within the biofilms but indicated that community diversity was low and varied between sampling dates. Biofilms were dominated by a few taxa, specifically Methylomonas, Acinetobacter, Mycobacterium, and Xanthomonadaceae, and the dominant taxa within the biofilms varied dramatically between sampling times. The drinking water characteristics most strongly correlated with bacterial community composition were concentrations of nitrate, ammonium, total chlorine and monochloramine, as well as alkalinity and hardness. Biofilms from the sampling date with the highest nitrate concentration were the most abundant and diverse and were dominated by Acinetobacter.     

The Usefulness of Various Numerical Methods for Assessing the Specific Effects of Pollution on Aquatic Biota

The data obtained during an extensive multidisciplinary investigation of a polluted harbor basin in Hamburg were analyzed in various ways in order to determine which statistical methods are easiest to interpret and permit the most accurate evaluations. The methods analyzed include the species deficit, index of species similarity, and saprobity index, as well as the more complex cluster and gradient analyses, which revealed a variety of relationships among the biotic communities at the various sampling sites and some ecological tolerance limits of various species. The shortcomings of numerical indices for characterizing water bodies are discussed. It is suggested that biostatistical methods are much more valuable for the empirical evaluation of data than for artificially classifying ecosystems.     

Natural versus anthropogenic charge in lakes: The role of the sediment record

Recent methodological developments permit the quantitative reconstruction of water chemistry variables from microfossil assemblages preserved in lake sediments. These reconstructions can be used to identify the extent and timing of disturbance to lake ecosystems. Combined with appropriate sampling strategies, lake sediments permit water chemistry variables and community rates of change to be estimated at a variety of timescales. Sediments predating major cultural impacts offer the possibility of inferring lake history before anthropogenic interference, and can, therefore, contribute to current debates about timescales of natural variance in lakes, as well as the response of lake communities to natural environmental perturbations. Such an approach has relevance to many contemporary environmental problems, e.g. acidification, eutrophication and climate change.     

Accuracy and precision of the topological mapping procedure for estimating within-tree populations of bark beetles

The accuracy and precision of the topological mapping procedure for estimating within-tree populations of bark beetles was investigated for a variety of different sampling conditions. Simulation techniques were used to define the mean and standard deviation of proportional errors encountered in estimation under different sampling intensities. The number of samples collected at a particular height and the vertical spacing between heights were varied. Information presented should aid in developing sampling plans for studies of bark beetle populations and will permit the reexamination and/or recovery of historical data sets on bark beetle populations.     

An integrative model of coupled water and solute exchange in the heart

Physiologists have devised many models for interpreting water and solute exchange data in whole organs, but the models have typically neglected key aspects of the underlying physiology to present the simplest possible model for a given experimental situation. We have developed a physiologically realistic model of microcirculatory water and solute exchange and applied it to diverse observations of water and solute exchange in the heart. Model simulations are consistent with the results of osmotic weight transient, tracer indicator dilution, and steady-state lymph sampling experiments. The key model features that permit this unification are the use of an axially distributed blood-tissue exchange region, inclusion of a lymphatic drain in the interstitium, and the independent computation of transcapillary solute and solvent fluxes through three different pathways.     

Unscreened Water-Diversion Pipes Pose an Entrainment Risk to the Threatened Green Sturgeon,Acipenser medirostris

Over 3,300 unscreened agricultural water diversion pipes line the levees and riverbanks of the Sacramento River (California) watershed, where the threatened Southern Distinct Population Segment of green sturgeon, Acipenser medirostris, spawn. The number of sturgeon drawn into (entrained) and killed by these pipes is greatly unknown. We examined avoidance behaviors and entrainment susceptibility of juvenile green sturgeon (35±0.6 cm mean fork length) to entrainment in a large (>500-kl) outdoor flume with a 0.46-m-diameter water-diversion pipe. Fish entrainment was generally high (range: 26–61%), likely due to a lack of avoidance behavior prior to entering inescapable inflow conditions. We estimated that up to 52% of green sturgeon could be entrained after passing within 1.5 m of an active water-diversion pipe three times. These data suggest that green sturgeon are vulnerable to unscreened water-diversion pipes, and that additional research is needed to determine the potential impacts of entrainment mortality on declining sturgeon populations. Data under various hydraulic conditions also suggest that entrainment-related mortality could be decreased by extracting water at lower diversion rates over longer periods of time, balancing agricultural needs with green sturgeon conservation.     

Developing optimum sample size and multistage sampling plans for Lobesia botrana (Lepidoptera: Tortricidae) larval infestation and injury in northern Greece

The purpose of this research was to quantify the spatial pattern and develop a sampling program for larvae of Lobesia botrana Denis and Schiffermüller (Lepidoptera: Tortricidae), an important vineyard pest in northern Greece. Taylor's power law and Iwao's patchiness regression were used to model the relationship between the mean and the variance of larval counts. Analysis of covariance was carried out, separately for infestation and injury, with combined second and third generation data, for vine and half-vine sample units. Common regression coefficients were estimated to permit use of the sampling plan over a wide range of conditions. Optimum sample sizes for infestation and injury, at three levels of precision, were developed. An investigation of a multistage sampling plan with a nested analysis of variance showed that if the goal of sampling is focusing on larval infestation, three grape clusters should be sampled in a half-vine; if the goal of sampling is focusing on injury, then two grape clusters per half-vine are recommended.     

Use ofCladophora glomerata to monitor heavy metals in rivers

Methods were developed for the use ofCladophora glomerata to monitor heavy metal concentrations in flowing waters. At least under conditions without marked fluctuations in ambient metal concentration, there was no detectable difference in the metal concentrations of young plants between terminal 2-cm lengths of filament and whole plants. In order to establish the relationship between metal concentration in plant and that in water, 60 algal and water samples were analyzed from sites in northern England for Fe, Cu, Zn, Cd and Pb. Other environmental variables were measured at the time in order to assess their influence on metal accumulation. There were highly significant correlations for each of the five metals between concentrations in alga and water. The regression equations relating metal in alga to metal in water permit an unknown environmental metal concentration to be estimated from the algal concentration. Multiple stepwise regression analyses were used to indicate environmental factors which may influence metal accumulation; for instance, Fe appears to have a positive influence on Cu accumulation. In generalCladophora accumulates much less metal than bryophytes, but the slope relating metal in alga to metal in water is steeper, particularly for Pb. This means thatCladophora is especially useful where there is a need for a sensitive indicator of differences between sites or sampling occasions.     

The association between distance to water pipes and water bodies positive for anopheline mosquitoes (Diptera: Culicidae) in the urban community of Malindi, Kenya.

The increasing risk of mosquito-borne diseases in African urban environments has been partly attributed to failed planning and resource underdevelopment. Though engineered systems may reduce mosquito proliferation, there are few studies describing this relationship. This study investigates how engineered systems such as roads and piped water systems affect the odds of anopheline immatures (i.e., larvae and pupae) occurring in water bodies located in Malindi, Kenya. Anopheles gambiae s.s. (Giles), An. arabiensis (Patton), and An. merus (Dointz) were identified in urban Malindi, with Anopheles gambiae s.s. being the predominant species identified. The Breslow-Day test was used to explore interactions among independent variables. Logistic regression was used to test whether water bodies positive for anopheline immatures are associated with engineered systems, while controlling for potential confounding and interaction effects associated with urban water body characteristics. Water bodies more than 100 m from water pipes were 13 times more likely to have anopheline immatures present, compared to water bodies that were less than 100 m from water pipes (OR = 13.54, 95% CI: 3.15-58.23). Roads were not significantly associated with water bodies positive for anopheline immatures. Statistical interaction was detected between water body substrate type and distance to water pipes. This study provides insight into how water pipes influence the distribution of water bodies positive with immature anophelines in urban environments.     

A light trap design for stratum-specific sampling of reef fish larvae

The vertical distribution of late stage reef fish larvae may potentially influence their dispersal, recruitment success and energetic expenditure during the recruitment process. To date, methods of examining the vertical distribution of reef fish larvae either under-sample late stage individuals, or are incapable of discretely sampling the water column. The aim of this study was to develop a light trap able to sample a narrow depth range enabling fine scale patterns of vertical distribution to be examined. The experimental traps radiated light in a relatively narrow beam with a maximum vertical angle of radiation of 7.5°, indicating that the traps could be placed 4.8 m apart and still sample discrete depth strata. Their catch efficiency was similar to conventional light traps, indicating that they are adequate sampling units. Preliminary data showed that most families are more abundant near the surface, although many have significant numbers of individuals lower in the water column. Some families (inc. Apogonidae) occurred in higher abundance at greater depths. Our experimental light traps permit increased resolution of the vertical distribution of late stage larval reef fishes in the field.     

Bioluminescence characteristics of Lake Shira water

Bioluminescence bioassays based on luminous bacteria (Photobacterium phosphopreum) and coupled enzyme system NADH-FMN-oxidoreductase-luciferase were adapted for monitoring the saline-water conditions of Lake Shira (Khakasia, Siberia). The differences in bioluminescence responses have been found to be related to the salt composition and the oxidation-reduction properties of water. Bioluminescent kinetics parameters, which are mostly sensitive to pollution under conditions of saline water, have been observed. The enzymatic system in the presence of 1,4-benzoquinone are shown to be more sensitive to redox characteristics of the salt water than this in the absence of 1,4-benzoquinone. 1,4-benzoquinone should be applied for the preparation of a model solution for the monitoring of redox properties of the salt water. Using this technique, the results of bioluminescence analysis are used to construct a heterogeneity map that characterizes the spatial and temporal water quality of lake Shira. A partial map was based on the bioluminescence characteristics of water samples taken along the shoreline, sampling stations in the different places and in different depths of the lake. It has been demonstrated that the bioluminescence assay measurements must be done within two hours after the sampling time.     

Retrieval of Concentrated and Undecompressed Microbial Populations from the Deep Sea

A device for sampling at depths of up to 6,000 m is described in which 3 liters of seawater is concentrated over a Nucleopore filter to about 13 ml and retrieved under in situ pressure and temperature. Subsamples can be withdrawn into transfer units that are equipped with individual gas accumulators for preventing loss of pressure during prolonged periods of storage. Transfer of samples or sample portions into sterile medium contained in pre-pressurized incubation vessels and continued subsampling therefrom permit time course experiments for the study of natural populations of deep-sea microorganisms in the absence of decompression. A test experiment with a water sample from a depth of 2,600 m supplemented with radioactively labeled Casamino Acids showed reduced rates of substrate incorporation and respiration as compared with data from a decompressed control. The barotolerance observed in this study was characterized by reduced, rather than equal, activities recorded at elevated pressures as compared with 1-atm controls.     

An impact of drainage system on soil water conditions at Lidzbark Warminski experimental site

The aim of this study is to evaluate the impact of a drainage system on soil water conditions in a loam soil compared to that in undrained clay loam soil under various topographic conditions. The soils are located on a sloping area at Lidzbark Warminski experimental site (Poland) with well surface water outflow conditions and used as a pasture. The loam soil was drained with ceramic drainage pipes with an average drain spacing of 14 m and an average drain depth of 0.9 m, while the clay loam soil profile was not drained. The research was conducted during the period from 1999 to 2005. Ground water level as well as soil moisture content were measured monthly for both soil profiles. Meteorological conditions (precipitation and data for calculation of reference evapotranspiration) were also recorded. The results obtained show that in the loam soil (drained site) water level is on average 42 cm higher compared to that in the clay loam soil (not drained site). In both soils the amplitude of the ground water level changes was relatively high and exceeds 300 cm. In the drained loam soil, the water level position exceeded the depth of the drainage system in very wet, wet and average years. Under wet meteorological conditions the increase in ground water levels in the clay loam soil was slower than in the loam soil.     

Are There Health Risks from the Migration of Chemical Substances from Plastic Pipes into Drinking Water? A Review

Plastic pipes used to convey hot and cold drinking water are synthetic polymers containing numerous additives that enhance durability, impact strength, and toughness, and resist material degradation. Although some research studies have been conducted to evaluate the type and levels of chemical substances migrating from polymeric materials into drinking water, the potential adverse health effects associated with these compounds in potable water have not been described. This review evaluates the literature on the occurrence of regulated and unregulated substances in drinking water related to the use of plastic pipes, characterizes potential health hazards, and describes uncertainties associated with human health and exposure in need of further research. Of particular public health concern is the potential for sensitive populations to be exposed to short-term elevations in leachates during critical periods, and for co-occurring leachates targeting the same organ(s) and/or sharing a common mode of toxic action to have additive or synergistic effects. Contaminants are measured in the distribution system, not at the tap where human exposure actually occurs. For increased health protection, it is important to identify compounds that migrate from plastic pipes into drinking water and to better quantify human exposures and health hazards to these substances and their degradates.     

Methods for repetitive measurements of multiple hematological parameters in individual rats

Methods have been developed which permit frequent repetitive blood sampling of rats without perturbing physiological parameters of interest. These techniques allow a comprehensive hematological study over several weeks, in individual rats, thus permitting full documentation of selected parameters during growth and development.     

Drip irrigation biofouling with treated wastewater: bacterial selection revealed by high-throughput sequencing

Clogging of drippers due to the development of biofilms weakens the advantages and impedes the implementation of drip irrigation technology. The objective of this study was to characterise the bacterial community of biofilms that develop in a drip irrigation system supplied with treated wastewater. High-throughput sequencing of 16S rRNA gene amplicons indicated that the bacterial community composition differed between drippers and pipes, mainly due to changes in the abundance of the genus Aquabacterium. Cyanobacteria were found to be involved in the biological fouling of drippers. Moreover, bacterial genera including opportunistic pathogenic bacteria such as Legionella and Pseudomonas were more abundant in dripper and pipe biofilms than in the incoming water. Some genera such as Pseudomonas were mostly recovered from drippers, while others (ie Bacillus, Brevundimonas) mainly occurred in pipes. Variations in the hydraulic conditions and properties of the materials likely explain the shift in bacterial communities observed between pipes and drippers.