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Biomedical subjects

Willy Verstraete

Publications and source records attributed to Willy Verstraete.

At least 91 records · Page 5Linked to original sources

Repression of Clostridium population in young broiler chickens after administration of a probiotic mixture.

The influence of the administration of a mixed probiotic on zootechnical performances and the microbial composition of the feces of young broilers, up 42 days old, was investigated. In a six weeks trial the feed was supplemented with 1(w/w)% of a mixture of five potentially probiotic strains belonging to the genera Lactobacillus and Enterococcus. No effect on the weight of the broilers was observed. Plate counting of fecal samples showed a decrease in the Clostridium population with more than 4 10log units after 5 days and 2 10log units after 14 days of treatment. The DGGE pattern obtained after nested PCR, revealed a significant shift in the Lactobacillus population. This study shows that the application of probiotics in the feed of broiler chickens considerably lowers the Clostridium population in broilers, hereby also lowering the risk of spreading in the housing through fecal contamination.

Animal Feed↗

Dimethyl sulfide removal from synthetic waste gas using a flat poly(dimethylsiloxane)-coated composite membrane bioreactor.

The reduction of volatile organic sulfur emissions should be completely as they cause odor nuisance, even when they are emitted in very small amounts. In general, biofilters are applied for odor reduction, but their operational control is limited. A new biotechnique for the treatment of complex emissions is the use of membranes integrated within bioreactors (MBRs). In this study, the reactor performance of MBRs for removal of dimethyl sulfide (DMS) as a model compound is presented. Composite membranes with a poly(dimethylsiloxane) (PDMS)-coating layer were used. The MBRs were inoculated with the sulfur-degrading culture Hyphomicrobium VS or a suspension of Hyphomicrobium VS, ammonium-oxidizing bacteria (AOB), and nitrite-oxidizing bacteria (NOB) to colonize the PDMS-coating layer. Although inoculation with AOB and NOB might give rise to competition for space on the membrane, their presence in the MBRs appeared to be positive as they co-oxidize DMS. Dimethyl sulfide elimination depended on the inoculum type, DMS inlet concentration, gas residence time, and membrane polymer. For equal loading rates, the elimination capacity (EC) increased at larger gas residence times and inlet concentrations. The maximum EC obtained with the MBRs was 4.8 kg of DMS x m(-3) x d(-1). This value is higher than any reported figure for biofilters and biotrickling filters.

Air Pollution↗

Release of angiotensin I converting enzyme (ACE) inhibitory activity during in vitro gastrointestinal digestion: from batch experiment to semicontinuous model.

Gastrointestinal digestion is of major importance in the bioavailability of angiotensin I converting enzyme (ACE) inhibitory peptides, bioactive peptides with possible antihypertensive effects. In this study, the conditions of in vitro gastrointestinal digestion leading to the formation and degradation of ACE inhibitory peptides were investigated for pea and whey protein. In batch experiments, the digestion simulating the physiological conditions sufficed to achieve the highest ACE inhibitory activity, with IC(50) values of 0.076 mg/mL for pea and 0.048 mg/mL for whey protein. The degree of proteolysis did not correlate with the ACE inhibitory activity and was always higher for pea than whey. In a semicontinuous model of gastrointestinal digestion, response surface methodology studied the influence of temperature and incubation time in both the stomach and small intestine phases on the ACE inhibitory activity and degree of proteolysis. For pea protein, a linear model for the degree of proteolysis and a quadratic model for the ACE inhibitory activity could be constituted. Within the model, a maximal degree of proteolysis was observed at the highest temperature and the longest incubation time in the small intestine phase, while maximal ACE inhibitory activity was obtained at the longest incubation times in the stomach and small intestine phase. These results show that ACE inhibitory activity of pea and whey hydrolysates can be controlled by the conditions of in vitro gastrointestinal digestion.

Angiotensin-Converting Enzyme Inhibitors↗

A novel approach to calcium removal from calcium-rich industrial wastewater.

Calcium-rich wastewater is a problem for industries due to calcification during downstream processing. The potential for Ca2+ removal from industrial wastewater through ureolytic microbiological carbonate precipitation was investigated for the first time. Batch experiments were used to determine feasible urea concentrations and hydraulic retention times. These results were applied in a semi-continuous reactor system, where the emphasis was placed on the development of a calcifying sludge. Calcium removal in excess of 90% was achieved throughout the experimental period, while the effluent pH remained at a reasonable level.

Calcium↗

Identification and quantification of nitrogen removal in a rotating biological contactor by 15N tracer techniques.

High autotrophic nitrogen removal rates of 858mg NL(-1) day(-1) or 1.55g Nm(-2) day(-1) were obtained in a lab-scale rotating biological contactor treating an ammonium rich influent. It was postulated that ammonium was removed as dinitrogen gas by a sequence of aerobic ammonium oxidation to nitrite taking place in the outer biofilm layer and anaerobic ammonium oxidation with nitrite as electron acceptor occuring in the deeper biofilm layer. Chemical evidence for anaerobic ammonium oxidation within intact biofilm sludge from a lab-scale rotating biological contactor could be provided, without direct identification of responsible organisms catalysing this reaction. 15N tracer techniques were used for identification and quantification of nitrogen transformations. In batch tests with biofilm sludge at dissolved oxygen concentrations lower than 0.1mgL(-1), ammonium and nitrite did react in a stoichiometric ratio of 1:1.43 thereby forming dinitrogen. 15N isotope dilution calculations revealed that anaerobic ammonium oxidation was the major nitrogen transformation leading to concomitant ammonium and nitrite removal. Isotopic analysis of the produced biogas showed that both ammonium-N and nitrite-N were incorporated in N(2).

Bacteria, Anaerobic↗

Test for assessing shear sensitivity of activated sludge flocs: a feasibility study.

Sludge deflocculation can cause, like excessive growth of filamentous bacteria, activated sludge wastewater treatment failure. Yet, unlike the latter cause, there exists no widely accepted tool to assess the flocculation level of activated sludge and to predict sludge deflocculation. In this study, a test procedure is proposed to assess the sensitivity of activated sludge flocs to shear. The test consists in subjecting sludge sample to a shear treatment followed by a sludge volume measurement (SV(30)), with reference to a control. The ratio (%) obtained by dividing the SV(30) of the sheared sample by the SV(30) of the control is used to express the shear sensitivity. In a first series of experiments using two types of sludge, the test was shown to be able of ranging sludge samples in a correct order of shear sensitivity. Applying this so-called shear-SV test procedure to a series of sludge samples allowed to distinguish amorphous sludges (SV(30) ratio> or =100%) from normal sludges (SV(30) ratio<100%). The test was shown to be repeatable and simple. It requires only basic laboratory equipment and implies minimum cost. It can be used as framework for the assessment of activated sludge shear sensitivity in practice.

Feasibility Studies↗

Removal of PCBs from wastewater using fly ash.

Liquids and sludges containing polychlorinated biphenyls (PCBs) can be treated to concentrate the PCBs in a solid residue. The latter can then be handled to destroy the PCBs. A study on sorption kinetics of PCBs on fly ash was conducted in controlled batch systems. TCB and HeCB are removed at 25 degrees C by adsorption on fly ash up to 97% at pH 7, with an adsorbent dose of 5 g/l. An examination of the thermodynamic parameters shows that the adsorption of TCB and HeCB by fly ash is a process occurring spontaneously at ambient conditions. Activation energies for the sorption process ranged between 5.6 and 49.1 kJ/mol. It was observed that the rate at which TCB and HeCB are adsorbed onto fly ash showed a diffusion limitation. The uptake rate of TCB and HeCB increases with increasing initial concentration and gradually tends to a constant value. A decrease in the adsorption of TCB and HeCB was observed when interfering ions and other PCB congeners were present. Changing the pH in the aqueous solution from 2 to 10 had no effect on the adsorption process. Overall, fly ash can be used for an efficient removal of PCBs from several aqueous solutions.

Adsorption↗

Impact of iron salts on activated sludge and interaction with nitrite or nitrate.

Iron salts are often used in activated sludge treatment plants as coagulants or to improve reactor performance. Previous studies have indicated that iron itself has an impact on the activated sludge process. However, the interaction of iron with nitrite or nitrate present in the sludge has received little attention. In this research, the influence of addition of Fe(II) or Fe(III), alone or together with NO(2)(-) or NO(3)(-) on bench-scale activated sludge reactors was examined. Large differences were established between the distinct treatments, regarding reactor performance, sludge characteristics as well as microbial community. Ferric iron was more detrimental than ferrous iron. In some cases, nitrite was found to enhance inhibitory effects of the added iron, whereas nitrate had more a neutralizing effect. It was found that precipitation of phosphate by the iron was not responsible for the observed inhibition. Decrease in pH upon formation of iron hydroxides and the impairment of the floc structure could partially explain the toxicity of the iron dosages. The formation of toxic nitrogen oxides, such as nitric oxide, can also be of importance. The observed positive effect of nitrate on the floc activity is of interest and warrants further elucidation.

Biodegradation, Environmental↗

Growth-substrate dependent dechlorination of 1,2-dichloroethane by a homoacetogenic bacterium.

A rod shaped, gram positive, non sporulating Acetobacterium strain was isolated that dechlorinated 1,2dichloroethane (1,2-DCA) to ethene at a dechlorination rate of up to 2 nmol Cl- min(-1) mg(-1) of protein in the exponential growth phase with formate (40 mM) as the substrate. Although with other growth substrates such as pyruvate, lactate, H2/CO2, and ethanol higher biomass productions were obtained, the dechlorination rate with these substrates was more than 10-fold lower compared with formate growing cells. Neither cell extracts nor autoclaved cells of the isolated Acetobacterium strain mediated the dechlorination of 1,2-DCA at significant rates. The addition of 1,2-DCA to the media did not result in increased cell production. No significant differences in corrinoid concentrations could be measured in cells growing on several growth-substrates. However, these measurements indicated that differences in corrinoid structure might cause the different dechlorination activity. The Acetobacterium sp. strain gradually lost its dechlorination ability during about 10 transfers in pure culture, probably due to undefined nutritional requirements. 16S rDNA analysis of the isolate revealed a 99.7% similarity with Acetobacterium wieringae. However, the type strains of A. wieringae and A. woodii did not dechlorinate 1,2-DCA.

Acetobacterium↗

A microbial fuel cell capable of converting glucose to electricity at high rate and efficiency.

A microbial fuel cell containing a mixed bacterial culture utilizing glucose as carbon source was enriched to investigate power output in relation to glucose dosage. Electron recovery in terms of electricity up to 89% occurred for glucose feeding rates in the range 0.5-3 g l(-1) d(-1), at powers up to 3.6 W m(-2) of electrode surface, a five fold higher power output than reported thus far. This research indicates that microbial electricity generation offers perspectives for optimization.

Bacteria, Anaerobic↗

Long-term effects of mineral versus organic fertilizers on activity and structure of the methanotrophic community in agricultural soils.

Agricultural practices, such as mineral nitrogen fertilization, have an impact on the soil's ability to oxidize methane, but little is known about the shifts in the methanotrophic community composition associated with these practices. Therefore, the long-term effect of both mineral (NH4NO3) and organic (manure and GFT-compost) fertilizer applications on the soil methanotrophic community activity and structure were investigated. Both high and low affinity methane oxidation rates were lower in the soil treated with mineral fertilizer compared to the other soils. An enhanced nitrate concentration was observed in the mineral fertilized soil but nitrate did not show a direct affect on the high affinity methane oxidation. In contrast, the low affinity methane oxidation was slowed down by increased nitrate concentrations, which suggests a direct effect of nitrate on low affinity methane oxidation. Denaturing gradient gel electrophoresis (DGGE) analysis of 16S rRNA gene fragments specific for methanotrophs revealed a distinct community between the mineral and organic fertilized soils as extra Type I methanotrophic bands (phylotypes) became visible in the organic fertilized soils. These phylotypes were not visible in the patterns of the added organic fertilizers suggesting an indirect effect of the organic fertilizers on the methanotrophic community. Additionally, a molecular analysis was performed after the low affinity methane oxidation test. The enhanced methane concentrations used in the test enriched certain low affinity methanotrophs in the organic fertilized soils but not in the mineral fertilized soil. Supporting the molecular and functional observations, fatty acids characteristic for methanotrophs were less abundant in the soil treated with mineral fertilizer compared to the soil treated with compost. In conclusion, the function and molecular and chemical composition of the methanotrophic community are all altered in soil fertilized with mineral fertilizer.

Alphaproteobacteria↗

Cell density related H2 consumption in relation to anoxic Fe(0) corrosion and precipitation of corrosion products by Shewanella oneidensis MR-1.

In the absence of oxygen, a protective H2 film is formed around an Fe(0) surface, inhibiting the electron flow from this surface. Our study of anoxic corrosion of Fe(0) beads revealed that, in the presence of Shewanella oneidensis MR-1, H2 removal and precipitation of Fe mineral particles on the cell surface are determining processes for corrosion. These two biologically mediated processes were governed by cell density. H2 removal by Shewanella oneidensis was detected at cell concentrations of 1.0 x 10(6) live cells ml-1 and higher and H2 was electron donor for denitrification of NO3-. The removal of the protective H2 layer from Fe(0) beads by Shewanella oneidensis, resulted in an increase of Fe release out of the Fe(0) beads from 153 +/- 25 mg l(-1) to 196 +/- 7 mg l-1 after 20 h. When the cell concentration exceeded 1.0 x 10(8) live cells ml-1, precipitation of iron minerals on the cell surface was characteristic for the greatest percentage of MR-1 cells, whereas micrometre-scale iron precipitates not associated with culturable cell biomass significantly decreased in number. Addition of supernatant of a corrosion assay with high cell concentration induced metabolic activity in a corrosion assay with low cell concentration, resulting in increased H2 consumption and Fe release from Fe(0) beads. Homoserine lactone-like molecules were detected in the supernatant by a bio-assay, suggesting the involvement of a quorum-sensing regulatory mechanism.

4-Butyrolactone↗

Bioaugmentation as a tool to protect the structure and function of an activated-sludge microbial community against a 3-chloroaniline shock load.

Bioaugmentation of bioreactors focuses on the removal of xenobiotics, with little attention typically paid to the recovery of disrupted reactor functions such as ammonium-nitrogen removal. Chloroanilines are widely used in industry as a precursor to a variety of products and are occasionally released into wastewater streams. This work evaluated the effects on activated-sludge reactor functions of a 3-chloroaniline (3-CA) pulse and bioaugmentation by inoculation with the 3-CA-degrading strain Comamonas testosteroni I2 gfp. Changes in functions such as nitrification, carbon removal, and sludge compaction were studied in relation to the sludge community structure, in particular the nitrifying populations. Denaturing gradient gel electrophoresis (DGGE), real-time PCR, and fluorescent in situ hybridization (FISH) were used to characterize and enumerate the ammonia-oxidizing microbial community immediately after a 3-CA shock load. Two days after the 3-CA shock, ammonium accumulated, and the nitrification activity did not recover over a 12-day period in the nonbioaugmented reactors. In contrast, nitrification in the bioaugmented reactor started to recover on day 4. The DGGE patterns and the FISH and real-time PCR data showed that the ammonia-oxidizing microbial community of the bioaugmented reactor recovered in structure, activity, and abundance, while the number of ribosomes of the ammonia oxidizers in the nonbioaugmented reactor decreased drastically and the community composition changed and did not recover. The settleability of the activated sludge was negatively influenced by the 3-CA addition, with the sludge volume index increasing by a factor of 2.3. Two days after the 3-CA shock in the nonbioaugmented reactor, chemical oxygen demand (COD) removal efficiency decreased by 36% but recovered fully by day 4. In contrast, in the bioaugmented reactor, no decrease of the COD removal efficiency was observed. This study demonstrates that bioaugmentation of wastewater reactors to accelerate the degradation of toxic chlorinated organics such as 3-CA protected the nitrifying bacterial community, thereby allowing faster recovery from toxic shocks.

Ammonia↗

Synergistic degradation of linuron by a bacterial consortium and isolation of a single linuron-degrading variovorax strain.

The bacterial community composition of a linuron-degrading enrichment culture and the role of the individual strains in linuron degradation have been determined by a combination of methods, such as denaturing gradient gel electrophoresis of the total 16S rRNA gene pool, isolation and identification of strains, and biodegradation assays. Three strains, Variovorax sp. strain WDL1, Delftia acidovorans WDL34, and Pseudomonas sp. strain WDL5, were isolated directly from the linuron-degrading culture. In addition, subculture of this enrichment culture on potential intermediates in the degradation pathway of linuron (i.e., N,O-dimethylhydroxylamine and 3-chloroaniline) resulted in the isolation of, respectively, Hyphomicrobium sulfonivorans WDL6 and Comamonas testosteroni WDL7. Of these five strains, only Variovorax sp. strain WDL1 was able to use linuron as the sole source of C, N, and energy. WDL1 first converted linuron to 3,4-dichloroaniline (3,4-DCA), which transiently accumulated in the medium but was subsequently degraded. To the best of our knowledge, this is the first report of a strain that degrades linuron further than the aromatic intermediates. Interestingly, the rate of linuron degradation by strain WDL1 was lower than that for the consortium, but was clearly increased when WDL1 was coinoculated with each of the other four strains. D. acidovorans WDL34 and C. testosteroni WDL7 were found to be responsible for degradation of the intermediate 3,4-DCA, and H. sulfonivorans WDL6 was the only strain able to degrade N,O-dimethylhydroxylamine. The role of Pseudomonas sp. strain WDL5 needs to be further elucidated. The degradation of linuron can thus be performed by a single isolate, Variovorax sp. strain WDL1, but is stimulated by a synergistic interaction with the other strains isolated from the same linuron-degrading culture.

Betaproteobacteria↗

Characterization of an autotrophic nitrogen-removing biofilm from a highly loaded lab-scale rotating biological contactor.

In this study, a lab-scale rotating biological contactor (RBC) treating a synthetic NH(4)(+) wastewater devoid of organic carbon and showing high N losses was examined for several important physiological and microbial characteristics. The RBC biofilm removed 89% +/- 5% of the influent N at the highest surface load of approximately 8.3 g of N m(-2) day(-1), with N(2) as the main end product. In batch tests, the RBC biomass showed good aerobic and anoxic ammonium oxidation (147.8 +/- 7.6 and 76.5 +/- 6.4 mg of NH(4)(+)-N g of volatile suspended solids [VSS](-1) day(-1), respectively) and almost no nitrite oxidation (< 1 mg of N g of VSS(-1) day(-1)). The diversity of aerobic ammonia-oxidizing bacteria (AAOB) and planctomycetes in the biofilm was characterized by cloning and sequencing of PCR-amplified partial 16S rRNA genes. Phylogenetic analysis of the clones revealed that the AAOB community was fairly homogeneous and was dominated by Nitrosomonas-like species. Close relatives of the known anaerobic ammonia-oxidizing bacterium (AnAOB) Kuenenia stuttgartiensis dominated the planctomycete community and were most probably responsible for anoxic ammonium oxidation in the RBC. Use of a less specific planctomycete primer set, not amplifying the AnAOB, showed a high diversity among other planctomycetes, with representatives of all known groups present in the biofilm. The spatial organization of the biofilm was characterized using fluorescence in situ hybridization (FISH) with confocal scanning laser microscopy (CSLM). The latter showed that AAOB occurred side by side with putative AnAOB (cells hybridizing with probe PLA46 and AMX820/KST1275) throughout the biofilm, while other planctomycetes hybridizing with probe PLA886 (not detecting the known AnAOB) were present as very conspicuous spherical structures. This study reveals that long-term operation of a lab-scale RBC on a synthetic NH(4)(+) wastewater devoid of organic carbon yields a stable biofilm in which two bacterial groups, thought to be jointly responsible for the high autotrophic N removal, occur side by side throughout the biofilm.

Anaerobiosis↗

Strain-specific ureolytic microbial calcium carbonate precipitation.

During a study of ureolytic microbial calcium carbonate (CaCO(3)) precipitation by bacterial isolates collected from different environmental samples, morphological differences were observed in the large CaCO(3) crystal aggregates precipitated within bacterial colonies grown on agar. Based on these differences, 12 isolates were selected for further study. We hypothesized that the striking differences in crystal morphology were the result of different microbial species or, alternatively, differences in the functional attributes of the isolates selected. Sequencing of 16S rRNA genes showed that all of the isolates were phylogenetically closely related to the Bacillus sphaericus group. Urease gene diversity among the isolates was examined by using a novel application of PCR-denaturing gradient gel electrophoresis (DGGE). This approach revealed significant differences between the isolates. Moreover, for several isolates, multiple bands appeared on the DGGE gels, suggesting the apparent presence of different urease genes in these isolates. The substrate affinities (K(m)) and maximum hydrolysis rates (V(max)) of crude enzyme extracts differed considerably for the different strains. For certain isolates, the urease activity increased up to 10-fold in the presence of 30 mM calcium, and apparently this contributed to the characteristic crystal formation by these isolates. We show that strain-specific calcification occurred during ureolytic microbial carbonate precipitation. The specificity was mainly due to differences in urease expression and the response to calcium.

Bacteria↗

Use of 16S-23S rRNA intergenic spacer region PCR and repetitive extragenic palindromic PCR analyses of Escherichia coli isolates to identify nonpoint fecal sources.

Despite efforts to minimize fecal input into waterways, this kind of pollution continues to be a problem due to an inability to reliably identify nonpoint sources. Our objective was to find candidate source-specific Escherichia coli fingerprints as potential genotypic markers for raw sewage, horses, dogs, gulls, and cows. We evaluated 16S-23S rRNA intergenic spacer region (ISR)-PCR and repetitive extragenic palindromic (rep)-PCR analyses of E. coli isolates as tools to identify nonpoint fecal sources. The BOXA1R primer was used for rep-PCR analysis. A total of 267 E. coli isolates from different fecal sources were typed with both techniques. E. coli was found to be highly diverse. Only two candidate source-specific E. coli fingerprints, one for cow and one for raw sewage, were identified out of 87 ISR fingerprints. Similarly, there was only one candidate source-specific E. coli fingerprint for horse out of 59 BOX fingerprints. Jackknife analysis resulted in an average rate of correct classification (ARCC) of 83% for BOX-PCR analysis and 67% for ISR-PCR analysis for the five source categories of this study. When nonhuman sources were pooled so that each isolate was classified as animal or human derived (raw sewage), ARCCs of 82% for BOX-PCR analysis and 72% for ISR-PCR analysis were obtained. Critical factors affecting the utility of these methods, namely sample size and fingerprint stability, were also assessed. Chao1 estimation showed that generally 32 isolates per fecal source individual were sufficient to characterize the richness of the E. coli population of that source. The results of a fingerprint stability experiment indicated that BOX and ISR fingerprints were stable in natural waters at 4, 12, and 28 degrees C for 150 days. In conclusion, 16S-23S rRNA ISR-PCR and rep-PCR analyses of E. coli isolates have the potential to identify nonpoint fecal sources. A fairly small number of isolates was needed to find candidate source-specific E. coli fingerprints that were stable under the simulated environmental conditions.

Animals↗

Stereoselective microbial dehalorespiration with vicinal dichlorinated alkanes.

The suspected carcinogen 1,2-dichloroethane (1,2-DCA) is the most abundant chlorinated C(2) groundwater pollutant on earth. However, a reductive in situ detoxification technology for this compound does not exist. Although anaerobic dehalorespiring bacteria are known to catalyze several dechlorination steps in the reductive-degradation pathway of chlorinated ethenes and ethanes, no appropriate isolates that selectively and metabolically convert them into completely dechlorinated end products in defined growth media have been reported. Here we report on the isolation of Desulfitobacterium dichloroeliminans strain DCA1, a nutritionally defined anaerobic dehalorespiring bacterium that selectively converts 1,2-dichloroethane and all possible vicinal dichloropropanes and -butanes into completely dechlorinated end products. Menaquinone was identified as an essential cofactor for growth of strain DCA1 in pure culture. Strain DCA1 converts chiral chlorosubstrates, revealing the presence of a stereoselective dehalogenase that exclusively catalyzes an energy-conserving anti mechanistic dichloroelimination. Unlike any known dehalorespiring isolate, strain DCA1 does not carry out reductive hydrogenolysis reactions but rather exclusively dichloroeliminates its substrates. This unique dehalorespiratory biochemistry has shown promising application possibilities for bioremediation purposes and fine-chemical synthesis.

Alkanes↗