Stefan Wuertz - Academia.edu (original) (raw)

Papers by Stefan Wuertz

The human health risks associated with multimedia exposure pathways of Ascaris lumbricoides (A. l... more The human health risks associated with multimedia exposure pathways of Ascaris lumbricoides (A. lumbricoides) are poorly understood including use of surface water contaminated with untreated wastewater in agriculture, consumption of uncooked raw vegetables irrigated with polluted stream-water, and incidental ingestion of fecally contaminated water by recreators. We analyzed a total of 65 water samples for the presence of helminthes collected from 5 monitoring locations in Salta City over a 13-month period on the Arias-Arenales river in the northwestern region of Salta province in Argentina. The average density of A. lumbricoides in the surface water was 5 eggs/liter. A. lumbricoides was detected in 35 of the 65 samples with a maximum concentration of 28 eggs/L. The risk of infection from accidental recreational ingestion of water was highest in children (1.31 x 10 -4 ) compared to the adults (6.47 x 10 -5 ) and secondary recreators (6.50 x 10 -6 ) whereas the annual probability of m...

Environmental Science & Technology, 2013

Journal of Bacteriology, 2000

The linked resistance to nickel and cobalt of Ralstonia eutropha-like strain CH34 (Alcaligenes eu... more The linked resistance to nickel and cobalt of Ralstonia eutropha-like strain CH34 (Alcaligenes eutrophus CH34) is encoded by the cnr operon, which is localized on the megaplasmid pMOL28. The regulatory genes cnrYXH have been cloned, overexpressed, and purified in Escherichia coli. CnrY fractionated as a 10.7-kDa protein in in vitro translation assays. CnrX, a periplasmic protein of 16.5 kDa, was overproduced and purified as a histidine-tagged fusion protein in E. coli. His-CnrX was found to posses a secondary structure content rich in alpha-helical and beta-sheet structures. CnrH, a sigma factor of the extracytoplasmic function family, was purified as an N-terminally histidine-tagged fusion. In gel shift mobility assays, His-CnrH, in the presence of E. coli core RNA polymerase enzyme, could retard at least two different promoter DNA targets, cnrYp and cnrHp, localized within the cnrYXH locus. These promoters and their transcription start sites were confirmed by primer extension. Purified His-CnrX did not inhibit the DNA-binding activity of His-CnrH and is therefore unlikely to be an anti-sigma factor, as previously hypothesized (EMBL M91650 description entry). To study the transcriptional response of the regulatory locus to metals and to probe promoter regions, transcriptional fusions were constructed between fragments of cnrYXH and the luxCDABE, luciferase reporter genes. Nickel and cobalt specifically induced the cnrYXH-luxCDABE fusion at optimal concentrations of 0.3 mM Ni 2؉ and 2.0 mM Co 2؉ in a noncomplexing medium for metals. The two promoter regions P Y (upstream cnrY) and P H (upstream cnrH) were probed and characterized using this vector and were found to control the nickelinducible regulatory response of the cnr operon. The cnrHp promoter was responsible for full transcription of the cnrCBA structural resistance genes, while the cnrYp promoter was necessary to obtain metal-inducible transcription from the cnrHp promoter. The zinc resistance phenotype (ZinB) of a spontaneous cnr mutant strain, AE963, was investigated and could be attributed to an insertion of IS1087, a member of the IS2 family of insertion elements, within the cnrY gene.

Proceedings of the Water Environment Federation, 2007

ABSTRACT

Water research, Jan 20, 2015

Bacteroidales and viruses were contemporaneously measured during dry and wet weather conditions a... more Bacteroidales and viruses were contemporaneously measured during dry and wet weather conditions at a watershed-scale in a semi-arid watershed impacted by a mixture of agricultural runoff, municipal wastewater effluent and municipal runoff. The results highlight the presence of municipal wastewater effluent as a confounding factor for microbial source tracking (MST) studies, and thus data were segregated into groups based on whether they were impacted by wastewater effluent. In semi-arid environments such as the Calleguas Creek watershed, located in southern California, the relative contribution of municipal wastewater effluent is dependent on hydrology as storm events lead to conditions where agricultural and municipal stormwater dominate receiving waters (rather than municipal wastewater, which is the case during dry weather). As such, the approach to data segregation was dependent on hydrology/storm conditions. Storm events led to significant increases in ruminant- and dog-associa...

Applied and environmental microbiology, 2014

Genetic exchange by natural transformation is an important mechanism of horizontal gene transfer ... more Genetic exchange by natural transformation is an important mechanism of horizontal gene transfer in biofilms. Thirty-two biofilm metrics were quantified in a heavily encapsulated Acinetobacter baylyi strain and a miniencapsulated mutant strain, accounting for cellular architecture, extracellular polymeric substances (EPS) architecture, and their combined biofilm architecture. In general, transformation location, abundance, and frequency were more closely correlated to EPS architecture than to cellular or combined architecture. Transformation frequency and transformant location had the greatest correlation with the EPS metric surface area-to-biovolume ratio. Transformation frequency peaked when EPS surface area-to-biovolume ratio was greater than 3 μm(2)/μm(3) and less than 5 μm(2)/μm(3). Transformant location shifted toward the biofilm-bulk fluid interface as the EPS surface area-to-biovolume ratio increased. Transformant biovolume was most closely correlated with EPS biovolume and ...

Journal of applied microbiology, 2014

The aim was to develop an optimized detachment method for separating Bacteroidales from sediments... more The aim was to develop an optimized detachment method for separating Bacteroidales from sediments to allow enumeration via PMA-qPCR. The effectiveness of four different detachment treatments in removing Bacteroides fragilis was compared as a function of time as well as in relation to Enterococcus faecalis and Escherichia coli as detected by cultivation and qPCR. Cells were inoculated into four sediments from sea water (SW) and freshwater (FW) beaches. Sediment samples were taken on days 1 and 7 and subjected to four different treatments for separation of micro-organisms. On day 1, the detachment treatments performed equally well in removing intact Bact. fragilis cells. In contrast, 7 days later the detachment treatment with Tween 80 and handshaking (TH) resulted in up to eightfold higher 16S rRNA gene concentrations of intact and total Bact. fragilis cells compared to other detachment treatments. Total Ent. faecalis cells based on the 23S rRNA gene were also preferentially recovered...

Biotechnology and bioengineering, 2014

We describe a novel procedure to estimate the net growth rate of biofilms on multiple substrates.... more We describe a novel procedure to estimate the net growth rate of biofilms on multiple substrates. The approach is based on diffusion-reaction mass balances for chemical species in a continuum biofilm model with reaction kinetics corresponding to a Double-Monod expression. This analytical model considers a heterogeneous biofilm with variable distributions of biofilm density, activity, and effective diffusivity as a function of depth. We present the procedure to estimate the effectiveness factor analytically and compare the outcome with values obtained by the application of a rigorous numerical computational method using several theoretical examples and a test case. A comparison of the profiles of the effectiveness factor as a function of the Thiele modulus, φ, revealed that the activity of a homogeneous biofilm could be as much as 42% higher than that of a heterogeneous biofilm, under the given conditions. The maximum relative error between numerical and estimated effectiveness facto...

Water research, 2010

The effects of toxins at ambient concentrations on microbial activity and community dynamics are ... more The effects of toxins at ambient concentrations on microbial activity and community dynamics are poorly understood. We operated 4 membrane bioreactors (MBRs) in parallel; two reactors were continuously exposed to the toxin 3-chloroaniline (3-CA) at environmentally relevant levels, representing 25% of the total chemical oxygen demand (COD; Total COD = 400 mg l(-1) d(-1)), and two reactors received no 3-CA. During the 70 d exposure to 3-CA the microbial communities never adapted as evidenced by a 48% and 14% reduction in COD and ammonia removal, respectively, compared to over 92% reduction for both measurements in the controls. The bacterial 16S rRNA gene was monitored using terminal restriction fragment length polymorphism (T-RFLP) analysis (n = 15 temporal grab samples per reactor) over the 70 d period. T-RFLP spectra analysis compared the rapid species turnover rate (STR) approach with the more computationally intensive non-metric multi-dimensional scaling (NMS) complemented with m...

Water research, 2007

We report the design and validation of new TaqMan((R)) assays for microbial source tracking based... more We report the design and validation of new TaqMan((R)) assays for microbial source tracking based on the amplification of fecal 16S rRNA marker sequences from uncultured cells of the order Bacteroidales. The assays were developed for the detection and enumeration of non-point source input of fecal pollution to watersheds. The quantitative "universal"Bacteroidales assay BacUni-UCD detected all tested stool samples from human volunteers (18 out of 18), cat (7 out of 7), dog (8 out of 8), seagull (10/10), cow (8/8), horse (8/8), and wastewater effluent (14/14). The human assay BacHum-UCD discriminated fully between human and cow stool samples but did not detect all stool samples from human volunteers (12/18). In addition, there was 12.5% detection of dog stool (1/8), but no cross-reactivity with cat, horse, or seagull fecal samples. In contrast, all wastewater samples were positive for the BacHum-UCD marker, supporting its designation as 100% sensitive for mixed-human source ...

Journal of water and health, 2006

The removal of target DNA by magnetic capture hybridization (MCH) from constituents inhibitory to... more The removal of target DNA by magnetic capture hybridization (MCH) from constituents inhibitory to amplification by polymerase chain reaction (PCR) was evaluated using Salmonella as the test pathogen. Hybrids were subjected to both conventional and quantitative real-time PCR (qPCR). When PCR inhibitors commonly found in water were added to the reaction, MCH-PCR increased the detection sensitivity on the order of 8 to 2,000-fold compared with the system using only PCR. To determine the selectivity of MCH for target DNA (Salmonella), different amounts of non-target DNA (Escherichia coli) were added to the qPCR reaction. The highest non-target DNA concentration interfered with the amplification by qPCR alone, while MCH-qPCR was unaffected. Average recovery of Salmonella DNA by MCH-qPCR was 31% using optimized buffers, washing solutions and enzymatic digestion. A recovery function was proposed in order to calculate the real cell number based on the measured value. Preliminary testing con...

Water science and technology : a journal of the International Association on Water Pollution Research, 2003

Confocal laser scanning microscopy was used to monitor the colonization pattern of the gfp-labele... more Confocal laser scanning microscopy was used to monitor the colonization pattern of the gfp-labeled derivative strain of Pseudomonas putida ATCC 17514 on fluorene and phenanthrene crystals. The in situ experiments showed that P. putida tends to grow directly on phenanthrene, forming a biofilm on accessible crystalline surfaces. On the other hand, no significant biofilm formation was observed in the presence of fluorene. The results obtained showed that substrate properties affected bacterial strategy regarding uptake.

Water science and technology : a journal of the International Association on Water Pollution Research, 2001

A method for extraction of extracellular polymeric substances (EPSs) with a dicyclohexyl-18-crown... more A method for extraction of extracellular polymeric substances (EPSs) with a dicyclohexyl-18-crown-6 ether was developed to determine levels of organic and inorganic contaminants sorbed to EPS. The crown ether selectively binds alkaline and alkaline earth metals but not heavy metals. The effectiveness of the extraction procedure was higher than that of 2 other methods tested and comparable with that of a method based on a cation exchange resin. On average it was possible to extract 20% of the TOC, 12% of the total protein content, and 4% of the total carbohydrate content of sludge or biofilm biomass. Metal sorption studies in activated sludge showed no influence of exposure time on the fractionation of metals within the biomass. Metals sorbed mostly to cellular material. In biofilms 12.2% of the cadmium and 9.1% of the zinc added was found in the EPS. In activated sludge EPS contained only 2.9% zinc. The distribution of metals within the biomass was dose dependent. The percentage of ...

Water Research, 2012

This review interprets highway runoff characterization studies performed on different continents.... more This review interprets highway runoff characterization studies performed on different continents. The results are synthesized to discuss the historical trends, first flush effects of pollutants, pollutant form as dissolved vs. particulate, and to identify surrogate water quality parameters. The information presented in this review showed that: (1) variability has been observed in all quality parameters from each continent and among continents; (2) with a few exceptions the variability seems to be within the expected range; (3) inconsistent monitoring data as well as inconsistent quality assurance and quality control measures were reported among studies, which may be partially responsible for variability of water quality results; (4) compared with historic data, the concentration of total Pb decreased exponentially, which can mostly be credited to leaded gasoline phase-out regulation; (5) first flush effects of pollutants based on concentration have been reported consistently (however, mass first flush effects for pollutants have been reported inconsistently compared with concentration first flush effect); (6) most metal pollutants and phosphorus are present in both the particulate and dissolved forms; and (7) strong correlations were observed between TSS, TDS, TOC and iron (Fe) and 13 other constituents and water quality parameters (turbidity, O&amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;G, TPH, DOC, TKN, EC, Cl, Cd, Cr, Cu, Ni, Pb, Zn).

Proceedings of the Water Environment Federation, 2011

Water Research, 2007

Microbial water quality a b s t r a c t Advances in microbial source tracking (MST) have largely ... more Microbial water quality a b s t r a c t Advances in microbial source tracking (MST) have largely been driven by the need to comply with water quality standards based on traditional indicator bacteria. Recently, a number of culture-independent, and library-independent methods based on polymerase chain reaction (PCR) have been gaining popularity among source trackers. However, only a limited number of these methods have been successfully used in field applications, primarily due to the fact that many of them are still being developed. In this critical outlook, we examine different viewpoints associated with the practical use of MST to identify critical research gaps, propose a priority-based timeline to address them, and outline emerging technologies that will likely impact the future of source tracking. We propose that it is necessary to consider each of these aspects in order to advance towards a unifying framework in source identification, so that fecal pollution monitoring can be reliably used for comprehensive environmental microbial monitoring, to develop risk assessment models, and to implement and validate adequate management practices.

Water Research, 2014

The relationship between microbial community structure and function has been examined in detail i... more The relationship between microbial community structure and function has been examined in detail in natural and engineered environments, but little work has been done on using microbial community information to predict function. We processed microbial community and operational data from controlled experiments with bench-scale bioreactor systems to predict reactor process performance. Four membrane-operated sequencing batch reactors treating synthetic wastewater were operated in two experiments to test the effects of (i) the toxic compound 3-chloroaniline (3-CA) and (ii) bioaugmentation targeting 3-CA degradation, on the sludge microbial community in the reactors. In the first experiment, two reactors were treated with 3-CA and two reactors were operated as controls without 3-CA input. In the second experiment, all four reactors were additionally bioaugmented with a Pseudomonas putida strain carrying a plasmid with a portion of the pathway for 3-CA degradation. Molecular data were generated from terminal restriction fragment length polymorphism (T-RFLP) analysis targeting the 16S rRNA and amoA genes from the sludge community. The electropherograms resulting from these T-RFs were used to calculate diversity indices - community richness, dynamics and evenness - for the domain Bacteria as well as for ammonia-oxidizing bacteria in each reactor over time. These diversity indices were then used to train and test a support vector regression (SVR) model to predict reactor performance based on input microbial community indices and operational data. Considering the diversity indices over time and across replicate reactors as discrete values, it was found that, although bioaugmentation with a bacterial strain harboring a subset of genes involved in the degradation of 3-CA did not bring about 3-CA degradation, it significantly affected the community as measured through all three diversity indices in both the general bacterial community and the ammonia-oxidizer community (α = 0.5). The impact of bioaugmentation was also seen qualitatively in the variation of community richness and evenness over time in each reactor, with overall community richness falling in the case of bioaugmented reactors subjected to 3-CA and community evenness remaining lower and more stable in the bioaugmented reactors as opposed to the unbioaugmented reactors. Using diversity indices, 3-CA input, bioaugmentation and time as input variables, the SVR model successfully predicted reactor performance in terms of the removal of broad-range contaminants like COD, ammonia and nitrate as well as specific contaminants like 3-CA. This work was the first to demonstrate that (i) bioaugmentation, even when unsuccessful, can produce a change in community structure and (ii) microbial community information can be used to reliably predict process performance. However, T-RFLP may not result in the most accurate representation of the microbial community itself, and a much more powerful prediction tool can potentially be developed using more sophisticated molecular methods.

Water Research, 2003

In this study we present a new approach to determine volumes, heterogeneity factors, and composit... more In this study we present a new approach to determine volumes, heterogeneity factors, and compositions of the bacterial population of activated sludge flocs by 3D confocal imaging. After staining the fresh flocs with fluorescein-isothiocyanate, 75 stacks of images (containing approx. 3000 flocs) were acquired with a confocal laser scanning microscope. The self-developed macro 3D volume and surface determination for the

Water Research, 2010

While reports on waterborne infections with Toxoplasma gondii are emerging worldwide, detection o... more While reports on waterborne infections with Toxoplasma gondii are emerging worldwide, detection of this zoonotic parasite in water remains challenging. Lack of standardized and quantitative methods for detection of T. gondii oocysts in water also limits research on the transport and fate of this pathogen through aquatic habitats. Here, we compare the ability of hollow-fiber ultrafiltration and capsule filtration to

Water Research, 2015

Host-associated Bacteroidales Quantitative PCR Persistence and survival Decay kinetics a b s t r ... more Host-associated Bacteroidales Quantitative PCR Persistence and survival Decay kinetics a b s t r a c t It is difficult to compare decay kinetics for genetic markers in an environmental context when they have been determined at different ambient temperatures. Therefore, we investigated the persistence of the host-associated genetic markers BacHum, BacCow and BacCan as well as the general Bacteroidales marker BacUni in both intact Bacteroidales cells and as total intracellular and extracellular marker DNA in controlled batch experiments at two temperatures using PMA-qPCR. Fecal Bacteroidales cells and DNA persisted longer at the lower temperature. Using the modified Arrhenius function to calculate decay constants for the same temperature, we then compared the decay of host-associated Bacteroidales cells and their DNA at 14 C in field-based flow-through microcosms containing human, cow, and dog feces suspended in freshwater or seawater and previously operated with an identical experimental design. The time for a 2-log reduction (T 99 ) was used to characterize host-associated Bacteroidales decay. Host-associated genetic markers as determined by qPCR had similar T 99 values in freshwater and seawater at 14 C when compared under both sunlight and dark conditions. In contrast, intact Bacteroidales cells measured by PMA-qPCR had shorter T 99 values in seawater than in freshwater. The decay constants of Bacteroidales cells were a function of physical (temperature) and chemical (salinity) parameters, suggesting that environmental parameters are key input variables for Bacteroidales survival in a predictive water quality model. Molecular markers targeting total Bacteroidales DNA were less susceptible to the variance of temperature, salinity and sunlight, implying that measurement of markers in both intact cells and DNA could enhance the predictive ScienceDirect j ou rnal h ome pag e: www.elsevier.com/loca te/watres w a t e r r e s e a r c h 7 0 ( 2 0 1 5 ) 2 0 5 e2 1 3 http://dx.