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Biomedical subjects

W Verstraete

Publications and source records attributed to W Verstraete.

At least 19 recordsLinked to original sources
PubMed2007

Performance and microbial analysis of defined and non-defined inocula for the removal of dimethyl sulfide in a biotrickling filter.

The performance and microbial communities of three differently inoculated biotrickling filters removing dimethyl sulfide (DMS) were compared. The biotrickling filters were inoculated with Thiobacillus thioparus TK-m (THIO), sludge (HANDS) and sludge + T. thioparus TK-m + Hyphomicrobium VS (HANDS++), respectively. The criteria investigated were length of the start-up period, the maximum elimination capacity, and the effects of intermittent loading rates, low pH, peak loading and very low loading rate on the DMS removal efficiency. The HANDS++ reactor exhibited the best performance considering all treatments. HANDS performed almost equally well as HANDS++, except during the determination of the EC(max), while THIO was generally the least efficient. During stable DMS loading at concentrations of 20 ppmv or lower, all reactors exhibited similar and high removal efficiencies (>99%). Denaturing gradient gel electrophoresis (DGGE) analysis showed the establishment of T. thioparus in the biofilm of all reactors, but not of Hyphomicrobium VS. Quantitative monitoring of the introduced bacterial strains was performed with a newly developed real-time PCR protocol. Initially, the inoculated strains were exclusively found in the reactors in which they were added. Afterwards, however, both strains developed in the biofilm of all three reactors, although T. thioparus attained higher cell densities than Hyphomicrobium. The presence of T. thioparus in THIO was related with the DMS loading rates that were applied, in the sense that intermittent DMS loading and very low DMS loading rates (0.5 ppmv) induced a decrease in gene copy numbers. Real-time PCR and DGGE both gave consistent results regarding the presence of Hyphomicrobium VS and Thiobacillus thioparus TK-m in the reactors. Only real-time PCR could be used to detect bacteria comprising of less than 1.4% of the total bacterial community ( approximately 10(5) copies ring(-1)).

Air Pollutants↗
PubMed2006

Contribution to the Detection and Identification of Oxidation Metabolites of Nonylphenol in Sphingomonas sp. strain TTNP3.

Sphingomonas sp. strain TTNP3 has been previously described as a bacterium that is capable of degrading the technical mixture of nonylphenol (NP) isomers and also the 4(3',5'-dimethyl-3'-heptyl)-phenol single isomer of NP. Until recently, 3,5-dimethyl-3-heptanol was the only reported metabolite of 4(3',5'-dimethyl-3'-heptyl)-phenol. A short time ago, the detection of an intracellular metabolite resulting from the oxidation of 4(3',5'-dimethyl-3'-heptyl)-phenol which was identified as 2(3,5-dimethyl-3-heptyl)-benzenediol has been reported. A decisive element for this identification was the occurrence of some slight differences with the two most probable metabolites i.e. 4(3',5'-dimethyl-3'-heptyl)-resorcinol and 4(3',5'-dimethyl-3'-heptyl)-catechol. These facts led us to hypothesise some NIH shift mechanisms explaining the formation of 2(3',5'-dimethyl-3'-heptyl)-benzenediol. In the present work, we describe the steps that led to the detection of these metabolites in the intracellular fraction of Sphingomonas sp. strain TTNP3. The formation of analogous intracellular metabolites resulting from the degradation of the technical mixture of NP is reported. To further elucidate these degradation products, studies were carried out with cells grown with 4(3',5'-dimethyl-3'-heptyl)-phenol as sole carbon source. The description of the syntheses of reference compounds, i.e. 4(3',5'-dimethyl-3'-heptyl)-resorcinol and 4(3',5'-dimethyl-3'-heptyl)-catechol and their comparative analyses with the intermediates of the degradation of 4(3',5'-dimethyl-3'-heptyl)-phenol are presented.

Chromatography, High Pressure Liquid↗
PubMed2006

Effluent quality of a conventional activated sludge and a membrane bioreactor system treating hospital wastewater.

Two lab scale wastewater treatment plants treating hospital wastewater in parallel were compared in terms of performance characteristics. One plant consisted of a conventional activated sludge system (CAS) and comprised an anoxic and aerobic compartment followed by a settling tank with recycle loop. The second pilot plant was a plate membrane bioreactor (MBR). The wastewater as obtained from the hospital had a variable COD (Chemical Oxygen Demand) ranging from 250 to 2300 mg l(-1). Both systems were operated at a similar hydraulic residence time of 12 hours. The reference conventional activated sludge system did not meet the regulatory standard for effluent COD of 125 mg l(-1) most of the time. Its COD removal efficiency was 88%. The plate MBR delivered an effluent with a COD value of 50 m g l(-1) or less, and attained an efficiency of 93%. The effluent contained no suspended particles. In addition, the MBR resulted in consistent operational parameters with a flux remaining around 8-10 l m(-2) h(-1) and a trans membrane pressure < 0.1 bar without the need for backwash or chemical cleaning. The CAS and the MBR system performed equally well in terms of TAN removal and EE2 removal. The CAS system typically decreased bacterial groups for about 1 log unit, whereas the MBR decreased these groups for about 3 log units. Enterococci were decreased below the detection limit in the MBR and indicator organisms such as fecal coliforms were decreased for 1.4 log units in the CAS system compared to a 3.6 log removal in the MBR.

Bioreactors↗
PubMed2006

Gnotobiotically grown aquatic animals: opportunities to investigate host-microbe interactions.

The culture of aquatic organisms is still hampered by the occurrence of unpredictable diseases in their early life stages, which are responsible for massive mortalities and considerable economic losses. A better understanding of the host-microbe interactions is certainly essential to develop effective solutions of disease control for the aquaculture industry. As demonstrated in terrestrial animals, the use of gnotobiotic systems (animals cultured in axenic conditions or with a known microflora) can be an excellent tool to extent the understanding of the mechanisms involved in host-microbe interactions and to evaluate new treatments of disease control. Several aquatic animals were cultured so far in germ-free conditions, such as fish, molluscs, crustaceans, rotifers, echinoderms, cnidarians, turbellarians, ascidians and echiurans. The aim of the present review is to recapitulate the findings obtained with gnotobiotic aquatic animals over the last decades, with special emphasis to the host-microbe interactions, as well as the perspectives for future research in this field. In addition, the procedures utilized to culture axenic aquatic animals and to verify contaminations are summarized, and the standardization of these procedures is proposed.

Animals↗
PubMed2006

Necrotrophic growth of Legionella pneumophila.

This study examined whether Legionella pneumophila is able to thrive on heat-killed microbial cells (necrotrophy) present in biofilms or heat-treated water systems. Quantification by means of plate counting, real-time PCR, and flow cytometry demonstrated necrotrophic growth of L. pneumophila in water after 96 h, when at least 100 dead cells are available to one L. pneumophila cell. Compared to the starting concentration of L. pneumophila, the maximum observed necrotrophic growth was 1.89 log units for real-time PCR and 1.49 log units for plate counting. The average growth was 1.57 +/- 0.32 log units (n = 5) for real-time PCR and 1.14 +/- 0.35 log units (n = 5) for plate counting. Viability staining and flow cytometry showed that the fraction of living cells in the L. pneumophila population rose from the initial 54% to 82% after 96 h. Growth was measured on heat-killed Pseudomonas putida, Escherichia coli, Acanthamoeba castellanii, Saccharomyces boulardii, and a biofilm sample. Gram-positive organisms did not result in significant growth of L. pneumophila, probably due to their robust cell wall structure. Although necrotrophy showed lower growth yields compared to replication within protozoan hosts, these findings indicate that it may be of major importance in the environmental persistence of L. pneumophila. Techniques aimed at the elimination of protozoa or biofilm from water systems will not necessarily result in a subsequent removal of L. pneumophila unless the formation of dead microbial cells is minimized.

Biofilms↗
PubMed2006

Genetic and genomic insights into the role of benzoate-catabolic pathway redundancy in Burkholderia xenovorans LB400.

Transcriptomic and proteomic analyses of Burkholderia xenovorans LB400, a potent polychlorinated biphenyl (PCB) degrader, have implicated growth substrate- and phase-dependent expression of three benzoate-catabolizing pathways: a catechol ortho cleavage (ben-cat) pathway and two benzoyl-coenzyme A pathways, encoded by gene clusters on the large chromosome (boxC) and the megaplasmid (boxM). To elucidate the significance of this apparent redundancy, we constructed mutants with deletions of the ben-cat pathway (the DeltabenABCD::kan mutant), the boxC pathway (the DeltaboxABC::kan mutant), and both pathways (the DeltabenABCDDelta boxABC::kan mutant). All three mutants oxidized benzoate in resting-cell assays. However, the DeltabenABCD::kan and DeltabenABCD DeltaboxABC::kan mutants grew at reduced rates on benzoate and displayed increased lag phases. By contrast, growth on succinate, on 4-hydroxybenzoate, and on biphenyl was unaffected. Microarray and proteomic analyses revealed that cells of the DeltabenABCD::kan mutant growing on benzoate expressed both box pathways. Overall, these results indicate that all three pathways catabolize benzoate. Deletion of benABCD abolished the ability of LB400 to grow using 3-chlorobenzoate. None of the benzoate pathways could degrade 2- or 4-chlorobenzoate, indicating that the pathway redundancy does not directly contribute to LB400's PCB-degrading capacities. Finally, an extensive sigmaE-regulated oxidative stress response not present in wild-type LB400 grown on benzoate was detected in these deletion mutants, supporting our earlier suggestion that the box pathways are preferentially active under reduced oxygen tension. Our data further substantiate the expansive network of tightly interconnected and complexly regulated aromatic degradation pathways in LB400.

Bacterial Proteins↗
PubMed2006

Microbial fuel cells for wastewater treatment.

Microbial fuel cells (MFCs) are emerging as promising technology for the treatment of wastewaters. The potential energy conversion efficiencies are examined. The rates of energy recovery (W/m3 reactor) are reviewed and evaluated. Some recent data relating to potato-processing wastewaters and a hospital wastewater effluent are reported. Finally, a set of process configurations in which MFCs could be useful to treat wastewaters is schematized. Overall, the MFC technology still faces major challenges, particularly in terms of chemical oxygen demand (COD) removal efficiency.

Anaerobiosis↗
PubMed2006

The treatment of hospital wastewater: an appraisal.

Hospitals discharge considerable amounts of chemicals and microbial agents in their wastewaters. Problem chemicals present in hospital wastewater belong to different groups, such as antibiotics, X-ray contrast agents, disinfectants and pharmaceuticals. Many of these chemical compounds resist normal wastewater treatment. They end up in surface waters where they can influence the aquatic ecosystem and interfere with the food chain. Humans are particularly exposed by the drinking water, produced from surface water. Microbial agents of special concern are multiresistant microbial strains. The latter are suspected to contribute to the spread of antibiotic resistance. In this paper, we will discuss the different approaches towards hospital wastewater treatment. The principle of uncoupling hospitals from public sewers warrants indepth evaluation by technologists and ecotoxicologists as well as public health specialists.

Anti-Bacterial Agents↗
PubMed2005

Culture independent detection of Sphingomonas sp. EPA 505 related strains in soils contaminated with polycyclic aromatic hydrocarbons (PAHs).

The Sphingomonas genus hosts many interesting pollutant-degrading strains. Sphingomonas sp. EPA505 is the best studied polycyclic aromatic hydrocarbon (PAH)-degrading Sphingomonas strain. Based on 16S rRNA gene sequence analysis, Sphingomonas sp. strain EPA505 forms a separate branch in the Sphingomonas phylogenetic tree grouping exclusively PAH-degrading isolates. For specific PCR detection and monitoring of Sphingomonas sp. EPA505 and related strains in PAH-contaminated soils, a new 16S rRNA gene-based primer set was designed. The new primer set was shown to be highly selective for Sphingomonas sp. strain EPA505 as it only amplified DNA from strain EPA505 and not from other tested Sphingomonas strains or soil bacteria not belonging to the Sphingomonas genus. Using DNA extracts of a variety of inoculated PAH-contaminated soils, the primer pair was able to detect EPA505 in concentrations as low as 10(2) cells per gram of soil. Applying the new primer set, 16S rRNA gene fragments which were 99-100% similar to the corresponding gene of strain EPA505 were amplified from four of five PAH-contaminated soils. On the other hand, no PCR products were obtained from any of five tested uncontaminated soils. The preferential presence of EPA505 related Sphingomonas strains in PAH-contaminated soils with very different contamination profiles and different origin suggests an important role of this type of Sphingomonas in the natural Sphingomonas community colonizing PAH-contaminated sites.

Biodegradation, Environmental↗
PubMed2005

Simultaneous quantification of differently glycosylated, acetylated, and 2,3-dihydro-2,5-dihydroxy-6-methyl-4H-pyran-4-one-conjugated soyasaponins using reversed-phase high-performance liquid chromatography with evaporative light scattering detection.

A novel method utilizing high-performance liquid chromatography (HPLC) with evaporative light scattering detection (ELSD) and electrospray ionisation mass spectrometry (ESI-MS) was developed for the analysis of soyasaponins, a divers group of triterpenic compounds with one or two sugar side chains, occurring in soy. Group A soyasaponins in different degrees of acetylation, as well as group B soyasaponins in both their 2,3-dihydro-2,5-dihydroxy-6-methyl-4H-pyran-4-one (DDMP)-conjugated and non-conjugated forms could be separated and quantified using authentic soyasaponin standards, in one single run. The method was tested by the determination of the soyasaponin content and composition of eight soygerm samples of different origin. Differences in the composition and the degree of acetylation of the group A soyasaponins were observed among these samples. The group B soyasaponins showed much less variability and they were mainly present in their DDMP-conjugated form.

Acetylation↗
PubMed2005

Operational and microbiological aspects of a bioaugmented two-stage biotrickling filter removing hydrogen sulfide and dimethyl sulfide.

A two-stage biotrickling filter was developed for removing dimethyl sulfide (DMS) and hydrogen sulfide (H2S). The first biotrickling filter (ABF) was inoculated with Acidithiobacillus thiooxidans and operated without pH control, while the second biotrickling filter (HBF) was inoculated with Hyphomicrobium VS and operated at neutral pH. High DMS elimination capacities were observed in the HBF (8.2 g DMS m(-3) h(-1) at 90% removal efficiency) after 2 days. Maximal observed elimination capacities were 83 g H2S m(-3) h(-1) (100% removal efficiency) and 58 g DMS m(-3) h(-1) (88% removal efficiency) for the ABF and the HBF, respectively. The influence of a decreasing empty bed residence time (120 down to 30 sec) and the robustness of the HBF towards changing operational parameters (low pH, starvation, and DMS and H2S peak loadings) were investigated. Suboptimal operational conditions rapidly resulted in lower DMS removal efficiencies, but recovery of the HBF was mostly obtained within a few days. The H2S removal efficiency in the ABF, however, was not influenced by varying operational conditions. In both reactors, microbial community dynamics of the biofilm and the suspended bacteria were investigated, using denaturing gradient gel electrophoresis (DGGE). After a period of gradual change, a stable microbial community was observed in the HBF after 60 days, although Hyphomicrobium VS was not the dominant microorganism. In contrast, the ABF biofilm community was stable from the first day and only a limited bacterial diversity was observed. The planktonic microbial community in the HBF was very different from that in the biofilm.

Acidithiobacillus thiooxidans↗
PubMed2005

Evaluation of nested PCR assays for the detection of Legionella pneumophila in a wide range of aquatic samples.

AIMS: To compare the sensitivities of two nested PCR assays for the detection of Legionella pneumophila to each other and to the plate counting technique (ISO 11731) in a wide range of aquatic samples. METHODS AND RESULTS: The nested PCR assay with the primer set LEG 225-LEG 858 revealed 56% of the 46 analysed aquatic samples as being positive for Legionella spp., while the primer set JFP-JRP yielded 98% positive samples. The detection was confirmed by sequencing the PCR products. These results are considerably higher than the result obtained with the plate counting technique (41%), indicating the higher sensitivity of PCR-based diagnostic methods. As the PCR assay with the LEG 225-LEG 858 primer set resulted in a lower number of positive samples, it is considered not sensitive enough for aquatic samples. Similar results for the respective primer sets were obtained for the detection of the species L. pneumophila, responsible for 90% of all human Legionella infections, in the aquatic samples analysed. Both microbial community analysis by PCR-denaturing gradient gel electrophoresis and the analysis of biotic and abiotic water quality parameters revealed no relation between L. pneumophila-positive and -negative samples and the physico-chemical and bacteriological characteristics of the aquatic samples. CONCLUSIONS: The results show the additional value of the PCR assay with the JFP-JRP primer set compared with the plate counting technique, as well as its applicability in a wide range of aquatic samples. SIGNIFICANCE AND IMPACT OF THE STUDY: This study shows the importance of comparing different primer sets for nested PCR assays for the detection of L. pneumophila in aquatic samples, as well as the lower sensitivity of the widely accepted plate counting technique (ISO 11731).

Base Sequence↗
PubMed2005

Failure of the ammonia oxidation process in two pharmaceutical wastewater treatment plants is linked to shifts in the bacterial communities.

AIMS: To investigate whether two different wastewater treatment plants (WWTPs) -- treating the same pharmaceutical influent -- select for a different bacterial and/or ammonia oxidizing bacterial (AOB) community. METHODS AND RESULTS: Molecular fingerprinting demonstrated that each WWTP had its own total bacterial and AOB community structure, but Nitrosomonas eutropha and N. europea were dominant in both WWTP A and B. The DNA and RNA analysis of the AOB communities revealed different patterns; so the most abundant species may not necessarily be the most active ones. Nitritation failures, monitored by chemical parameter analysis, were reflected as AOB community shifts and visualized by denaturing gradient gel electrophoresis (DGGE)-based moving window analysis. CONCLUSIONS: This research demonstrated the link between functional performance (nitritation parameters) and the presence and activity of a specific microbial ecology (AOB). Clustering and moving window analysis based on DGGE showed to be valuable to monitor community shifts in both WWTPs. SIGNIFICANCE AND IMPACT OF THE STUDY: This study of specific community shifts together with functional parameter analysis has potential as a tool for relating functional instability (such as operational failures) to specific-bacterial community shifts.

Ammonia↗
PubMed2005

Growth substrate- and phase-specific expression of biphenyl, benzoate, and C1 metabolic pathways in Burkholderia xenovorans LB400.

Recent microarray experiments suggested that Burkholderia xenovorans LB400, a potent polychlorinated biphenyl (PCB)-degrading bacterium, utilizes up to three apparently redundant benzoate pathways and a C(1) metabolic pathway during biphenyl and benzoate metabolism. To better characterize the roles of these pathways, we performed quantitative proteome profiling of cells grown on succinate, benzoate, or biphenyl and harvested during either mid-logarithmic growth or the transition between the logarithmic and stationary growth phases. The Bph enzymes, catabolizing biphenyl, were approximately 16-fold more abundant in biphenyl- versus succinate-grown cells. Moreover, the upper and lower bph pathways were independently regulated. Expression of each benzoate pathway depended on growth substrate and phase. Proteins specifying catabolism via benzoate dihydroxylation and catechol ortho-cleavage (ben-cat pathway) were approximately an order of magnitude more abundant in benzoate- versus biphenyl-grown cells at the same growth phase. The chromosomal copy of the benzoyl-coenzyme A (CoA) (box(C)) pathway was also expressed during growth on biphenyl: Box(C) proteins were approximately twice as abundant as Ben and Cat proteins under these conditions. By contrast, proteins of the megaplasmid copy of the benzoyl-CoA (box(M)) pathway were only detected in transition-phase benzoate-grown cells. Other proteins detected at increased levels in benzoate- and biphenyl-grown cells included general stress response proteins potentially induced by reactive oxygen species formed during aerobic aromatic catabolism. Finally, C(1) metabolic enzymes were present in biphenyl-grown cells during transition phase. This study provides insights into the physiological roles and integration of apparently redundant catabolic pathways in large-genome bacteria and establishes a basis for investigating the PCB-degrading abilities of this strain.

Aerobiosis↗
PubMed2005

Anaerobic digestion as a core technology in sustainable management of organic matter.

In the past decades, anaerobic digestion (AD) has steadily gained importance. However, the technology is not regarded as a top priority in science policy and in industrial development at present. In order for AD to further develop, it is crucial that AD profits from the current fuel issues emerging in the international arena. AD can provide low-cost treatment of sewage and solid domestic wastes, which represents a vast application potential that should be promoted in the developing world. Furthermore, the developments in the last decades in the domain of anaerobic microbiology and technology have generated some interesting niches for the application of AD, such as anaerobic nitrogen removal and the treatment of chlorinated organics. Recently, AD has also generated some serendipities, such as the use of AD in processes for sulphur and calcium removal and the coupling of AD with microbial fuel cells. The international developments in terms of bio-refineries and CO2-emission abatement are of crucial importance with respect to the impetus that AD will receive in the coming decade. There should be little doubt that by placing the focus of AD on the production of green energy and clean nutrients, the future of AD will be assured.

Bacteria, Anaerobic↗
PubMed2005

Continuous microbial fuel cells convert carbohydrates to electricity.

Microbial fuel cells which are operated in continuous mode are more suitable for practical applications than fed batch ones. Three influent types containing carbohydrates were tested, i.e. a glucose medium, a plant extract and artificial wastewater. The anode reactor compartment yielding the best results was a packed bed reactor containing graphite granules. While in non-mediated batch systems power outputs up to 479 W m(-3) of anode compartment could be attained; in continuous mode the power outputs were limited to 49W m(-3). Cyclic voltammetry was performed to determine the potential of the in-situ synthesized bacterial redox mediators. Addition of mediators with a potential similar to the bacterial potential did not significantly alter the MFC power output, indicating a limited influence of soluble mediators for continuous microbial fuel cells. Maximum coulombic and energy conversion efficiencies were, for the continuous microbial fuel cell operating on plant extract at a loading rate of 1 kg COD m(-3) of anode compartment per day, 50.3% and 26.0% respectively.

Acer↗
PubMed2004

A sensitivity study for the visualisation of bacterial weathering of concrete and stone with computerised X-ray microtomography.

Geologists and engineers recently have adopted computerised X-ray microtomography (microCT), a radiological imaging technique, for geological and petrophysical applications such as the assessment of sediment characteristics, CT-measurement of compressibility and compaction, multiphase flow studies, measurement of bulk density and moisture content, and of porosity and permeability. This study focuses on another application: the monitoring of biological weathering of natural building stones and concrete. Microbial activity as a determining factor in the deterioration process of building materials has a major economic impact. Because of its non-destructive character, microCT could be the ideal monitoring technique. With this technique, three-dimensional (3D) images of the entire inner structure of the material can be obtained, together with quantitative data. In depth changes of porosity of concrete and stone specimens due to bacterial weathering were assessed in this work. Also, porosity was visualised based on 3D data with homemade software. Scanning electron microscopy (SEM) images provided additional information and supported conclusions drawn from the X-ray microCT data. Resolution improvement will make the study of petrophysical aspects of physical weathering and/or biological deterioration processes of natural building stones and concrete a promising subject for further microCT-application.

Biodegradation, Environmental↗