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Biomedical subjects

W Verstraete

Publications and source records attributed to W Verstraete.

At least 145 records · Page 8Linked to original sources

Contact angle measurement and cell hydrophobicity of granular sludge from upflow anaerobic sludge bed reactors.

The contact angle, which is generally used to evaluate the hydrophobicities of pure bacterial strains and solid surfaces, was used to study mixed cell cultures of bacteria involved in anaerobic digestion. Previously published data and data from this study showed that most acidogens are hydrophilic (contact angle, <45(deg)) but most of the acetogens and methanogens isolated from granular sludge are hydrophobic (contact angle, >45(deg)). The hydrophobicities of mixtures of hydrophilic and hydrophobic cells were found to be linearly correlated with the cell mixing ratio. The hydrophobicities of cells present in effluents from upflow anaerobic sludge bed reactors which were treating different types of substrates were different depending on the reactor conditions. When the reactor liquid had a high surface tension, cells sloughing off from sludge granules, as well as cells present on the outer surfaces of the granules, were hydrophobic. Short-term batch enrichment cultures revealed that proteins selected for highly hydrophilic cells. Long-term in-reactor enrichment cultures revealed that sugars selected for hydrophilic acidogens on the surfaces of the granules, while fatty acids tended to enrich for hydrophobic methanogens. When linear alkylbenzenesulfonate was added, the cells on the surfaces of granules became more hydrophilic. Control tests performed with pure cultures revealed that there was no change in the surface properties due to linear alkylbenzenesulfonate; hence, the changes in the wash-out observed probably reflect changes in the species composition of the microbial association. A surface layer with moderate hydrophobicity, a middle layer with extremely high hydrophobicity, and a core with high hydrophobicity could be distinguished in the grey granules which we studied.

Journal Article↗

Granulation and sludge bed stability in upflow anaerobic sludge bed reactors in relation to surface thermodynamics.

Adhesion of bacteria involved in anaerobic consortia was investigated in upflow anaerobic sludge bed reactors and was related to surface thermodynamics. The adhesion of hydrophilic cells appeared to be enhanced at a low liquid surface tension ((gamma)(infLV)), while the adhesion of hydrophobic cells was favored at a high (gamma)(infLV). Growth in protein-rich growth media resulted in low granular biomass yields; addition of polycations, such as poly-l-lysine and chitosan, increased the (gamma)(infLV) and the granular biomass yield. On the basis of the results of activity tests and microbial counts with wash-out cells, we identified two types of structured granules that were related to the influence of (gamma)(infLV). In one type of granules, hydrophilic acidogens surrounded a more hydrophobic methanogenic association. These granules were selected at a low (gamma)(infLV) provided that carbohydrates were available as substrates. The other type of granules was selected at a high (gamma)(infLV); hydrophobic cells (i.e., methanogens) were predominant throughout these granules. The granules which had acidogens as solid-phase emulsifiers around a methanogenic association appeared to allow more stable reactor performance. Decreasing the (gamma)(infLV) in the reactor by adding trace amounts of a surfactant also increased reactor stability.

Journal Article↗

Survival and Plant Growth Promotion of Detergent-Adapted Pseudomonas fluorescens ANP15 and Pseudomonas aeruginosa 7NSK2.

Four detergents were tested as selective C sources for the plant growth-promoting rhizobacteria Pseudomonas aeruginosa 7NSK2 and Pseudomonas fluorescens ANP15. CO-720 (Igepal CO-720) or DOS (dioctyl sulfosuccinate), applied at 0.2% to the soil, increased the number of detergent-adapted, inoculated strains by almost 1.5 log units after 25 days, accounting for virtually the entire increase in total bacteria. The same dose of Tween 80 or N-laurylsarcosine, on the other hand, increased the indigenous populations by almost 2.5 log units, with only minor increases in the number of detergent-adapted inoculated strains. When CO-720 or DOS was initially supplied, the number of detergent-adapted 7NSK2 organisms was about 2 log units higher after 3 months of incubation than for the detergent-unadapted strain. This better survival resulted in a significantly higher root colonization of maize in a pot experiment with soil inoculation, with a significantly (P <= 0.05) higher shoot dry weight (18 to 33%). In a first field experiment with rhizosphere inoculation of 1-month-old maize plants, no effects on the height of two maize cultivars could be observed 1 month after inoculation. In a second field experiment, leaf and stem dry weights of yellow mustard and grass dry weight were increased in the treatments with seed and soil inoculation of the detergent-adapted 7NSK2 in combination with CO-720 application by, respectively, 7 to 8%, 19 to 23%, and 20 to 31%, although only the increases in grass dry weight were statistically significant at P <= 0.1. To some extent, 7NSK2 and DOS application also positively affected the mineral content of yellow mustard.

Journal Article↗

Exogenous isolation of mobilizing plasmids from polluted soils and sludges.

Exogenous plasmid isolation was used to assess the presence of mobilizing plasmids in several soils and activated sludges. Triparental matings were performed with Escherichia coli (a member of the gamma subgroup of the Proteobacteria) as the donor of an IncQ plasmid (pMOL155, containing the heavy metal resistance genes czc: Co, Zn, and Cd), Alcaligenes eutrophus (a member of the beta subgroup of the Proteobacteria) as the recipient, and indigenous microorganisms from soil and sludge samples as helper strains. We developed an assay to assess the plasmid mobilization potential of a soil ecosystem on the basis of the number of transconjugants obtained after exogenous isolations. After inoculation into soil of several concentrations of a helper strain (E. coli CM120 harboring IncP [IncP1] mobilizing plasmid RP4), the log numbers of transconjugants obtained from exogenous isolations with different soil samples were a linear function of the log numbers of helper strain CM120(RP4) present in the soils. Four soils were analyzed for the presence of mobilizing elements, and mobilizing plasmids were isolated from two of these soils. Several sludge samples from different wastewater treatment plants yielded much higher numbers of transconjugants than the soil samples, indicating that higher numbers of mobilizing strains were present. The mobilizing plasmids isolated from Gent-O sludge and one plasmid isolated from Eislingen soil hybridized to the repP probe, whereas the plasmids isolated from Essen soil did not hybridize to a large number of rep probes (repFIC, repHI1, repH12, repL/M, repN, repP, repT, repU, repW, repX). This indicates that in Essen soil, broad-host-range mobilizing plasmids belonging to other incompatibility groups may be present.

Journal Article↗

Fibrous dysplasia in the frontoethmoidal complex: diagnosis and surgical aspects.

Fibrous dysplasia of the paranasal sinuses is rare and often presents a diagnostic challenge. We describe 3 cases of fibrous dysplasia and ossifying fibroma involving the paranasal sinuses. After introduction of the term fibrous dysplasia into the literature in 1938 by Lichtenstein, many authors accepted fibrous dysplasia and ossifying fibroma as one and the same entity. In more recent years most investigators have considered the two lesions as separate and distinct clinical entities. A brief update of the clinical aspects, radiographic appearance, diagnosis and management of craniofacial fibro-osseous lesions is provided.

Adolescent↗

Development of a 5-step multi-chamber reactor as a simulation of the human intestinal microbial ecosystem.

A five-stage reactor was developed to simulate the gastro-intestinal microbial ecosystem of humans. The small intestine was simulated by a two-step "fill and draw" system, the large intestine by a three-step reactor. A representative supply medium was developed to support a microbial community resembling that of the human gastro-intestinal tract. The entire system was validated by monitoring fermentation fluxes and products, i.e. indicator bacterial groups, volatile fatty acids, enzymatic activities and headspace gases. The simulator was operated with varying concentrations and combinations of arabinogalactan, xylan, pectin, dextrins and starch. The resulting patterns of microbial diversity and activity were analysed and compared with data for in-vivo gastro-intestinal microbial communities as described in the literature and found to be representative.

Ammonia↗

Gene disruption in Lactobacillus plantarum strain 80 by site-specific recombination: isolation of a mutant strain deficient in conjugated bile salt hydrolase activity.

A chloramphenicol-resistance gene (cml) was introduced into the Lactobacillus plantarum gene encoding conjugated bile acid hydrolase (cbh) on a ColE1 replicon. This plasmid which is nonreplicative in Lactobacillus was used to transform L. plantarum strain 80. A homologous double cross-over recombination event resulted in replacement of the chromosomal cbh gene by the cml-containing cbh gene. The transformants obtained were unable to synthesize active conjugated bile acid hydrolase (Cbh). The Cbh- CmlR phenotype was stably maintained for more than 100 generations under nonselective conditions.

Amidohydrolases↗

An activated sludge-based biosensor for rapid IC50 estimation and on-line toxicity monitoring.

The RODTOX (Rapid Oxygen Demand and TOXicity tester), an activated sludge-based respirographic biosensor, is a device for on-line monitoring of the short-term biochemical oxygen demand (stBOD) and potential toxicity of incoming wastewater on the basis of on-line interpretation of respirograms resulting from pulse additions of either calibration substrate or sample. The principle of toxicity detection is based on the comparison of calibration respirograms before and after receiving a potential toxicant. In this paper, the results of the RODTOX as an on-line toxicity monitor are presented. In addition, a simple and fast procedure to estimate the IC50 of a toxicant has been developed, and its validity and good repeatability demonstrated. The performance of this procedure is compared with that of the Microtox test.

Biosensing Techniques↗

Prevalence of coaggregation reactions among chicken lactobacilli.

Interbacterial adherence was frequently encountered among chicken lactobacilli. Fourteen of 45 combinations involving nine adhering strains were shown to be coaggregative. The coadherence mechanism was mediated by complementary heat- and sonication-sensitive cell surface structures. It was shown that intrageneric adherence enabled lactobacilli to maintain higher numbers in fed-batch reactors simulating the gastrointestinal tract. The mechanism of coaggregation can substantially increase the colonization potential of lactobacilli in environments with short residence times.

Animals↗

Physicochemical characterization of the microbial Fe2+ chelator proferrorosamine from Pseudomonas roseus fluorescens.

The microbial chelating compound proferrorosamine A, produced by Pseudomonas roseus fluorescens, formed a complex with Fe2+ of which the apparent stability constant was found to be 10(23). The following order of increasing stability constants of metal complexes with proferrorosamine was established as: Ba2+, Ca2+, Mg2+, Mn2+ less than Hg2+ less than Zn2+ less than Pb2+ less than Co2+ less than Cu2+ congruent to Fe2+ less than Ni2+. Only Ni(2+)-proferrorosamine had a stability constant which was established as: Ba2+, Ca2+, Mg2+, Mn2+ less than Hg2+ less than Zn2+ less than Pb2+ less than Co2+ less than Cu2+ congruent to Fe2+ less than Ni2+. Only Ni(2+)-proferrorosamine had a stability constant which was ca 32 times higher than Fe(2+)-proferrorosamine. Because of the production of proferrorosamine the growth of Ps. roseus fluorescens was not inhibited in iron limiting media by the addition of 0.15 mmol/l of the weaker chemical Fe2+ chelator 2,2'-dipyridyl. This contrasted with the proferrorosamine-negative mutant K2 and Ps. stutzeri, which only produces Fe(3+)-chelating siderophores. Furthermore, it was found that proferrorosamine was able to dissolve Fe2+ from stainless steel. These results show that proferrorosamine is a strong and selective Fe2+ chelator which could be used as an alternative for the toxic 2,2'-dipyridyl to control lactic acid fermentations.

2,2'-Dipyridyl↗

Cloning and expression of a conjugated bile acid hydrolase gene from Lactobacillus plantarum by using a direct plate assay.

The conjugated bile acid hydrolase gene from the silage isolate Lactobacillus plantarum 80 was cloned and expressed in Escherichia coli MC1061. For the screening of this hydrolase gene within the gene bank, a direct plate assay developed by Dashkevicz and Feighner (M. P. Dashkevicz and S. D. Feighner, Appl. Environ. Microbiol. 53:331-336, 1989) was adapted to the growth requirements of E. coli. Because of hydrolysis and medium acidification, hydrolase-active colonies were surrounded with big halos of precipitated, free bile acids. This phenomenon was also obtained when the gene was cloned into a multicopy shuttle vector and subsequently reintroduced into the parental Lactobacillus strain. The cbh gene and surrounding regions were characterized by nucleotide sequence analysis. The deduced amino acid sequence was shown to have 52% similarity with a penicillin V amidase from Bacillus sphaericus. Preliminary characterization of the gene product showed that it is a cholylglycine hydrolase (EC 3.5.1.24) with only slight activity against taurine conjugates. The optimum pH was between 4.7 and 5.5. Optimum temperature ranged from 30 to 45 degrees C. Southern blot analysis indicated that the cloned gene has similarity with genomic DNA of bile acid hydrolase-active Lactobacillus spp. of intestinal origin.

Amidohydrolases↗

Determination of the mechanism of retrotransfer by mechanistic mathematical modeling.

Two mathematical models to elucidate the mechanism of retromobilization (or retrotransfer), that is, the ability of conjugative plasmids to mobilize genes into the cell containing the conjugative plasmid, were developed. This study deals with retromobilization of nonconjugative plasmids (Tra-Mob+). Plasmid transfer was modeled by two mass action models. The first is based on the hypothesis that retromobilization of the Tra-Mob+ vector occurs in one step, by means of the pilus formed by the Tra+ plasmid in the original host. In the second model, retromobilization is considered to be a two-step process involving two transfer events. The first step involves the transfer of the Tra+ plasmid from the recipient cell to the donor of the nonconjugative vector, and during the second encounter the nonconjugative vector is mobilized toward the recipient. Since the relationships between the number of transconjugants and the number of recipients for the two models are different, filter matings were performed for short time periods with different initial densities of the recipient population. Comparison of the numbers of transconjugants with the results of the mathematical equations confirmed the hypothesis that retromobilization is a one-step conjugation process.

Conjugation, Genetic↗

Detection and prevalence of Listeria monocytogenes in the agricultural ecosystem.

The sensitivity of four different enrichment procedures to detect Listeria monocytogenes in the presence of high levels of Streptococcus faecalis was investigated. Defined mixed cultures of Strep. faecalis and L. monocytogenes gave better results with one-stage enrichment techniques. For manure samples, however, two-stage enrichment techniques gave the best performance. The so-called cold enrichment techniques were found to be unsatisfactory for samples from natural environments. The following materials were examined for the presence of L. monocytogenes: fresh pig faeces (16% positive), fresh cattle faeces (20% positive), stored liquid manure (0% positive), manured soil samples (0% positive) and ground water samples (5% positive). After 3 weeks of storage L. monocytogenes could be detected in only one of the initially nine positive fresh faeces samples. Two months after inoculation of stored liquid pig manure, stored liquid cattle manure and soil with L. monocytogenes, this bacterium could not be traced in any of these materials. Radishes (Raphanus sativus) and carrots (Daucus carota), sown in soil inoculated with L. monocytogenes, were gathered after 3 months and examined for the presence of L. monocytogenes. Three of six radish samples were found to be positive. Remarkably, however, all carrot samples (six) were free of L. monocytogenes.

Animals↗

The Fe Chelator Proferrorosamine A Is Essential for the Siderophore-Mediated Uptake of Iron by Pseudomonas roseus fluorescens.

Pseudomonas roseus fluorescens produces, besides the Fe chelator proferrorosamine A, Fe -chelating compounds, called siderophores. The production of proferrorosamine A and siderophores by P. roseus fluorescens appears to be controlled in a similar way by the concentration of available iron and by the concentration of dissolved oxygen. The higher the concentration of iron available for the microorganism, the lower the production of both chelating compounds. However, the production of siderophores was much more sensitive to iron availability than was proferrorosamine A production. Proferrorosamine A and siderophores were only produced in minimal medium C if the concentration of dissolved oxygen ranged from 4.5 to 2.0 ppm. At higher or lower concentrations, none of the iron-chelating compounds were produced. Furthermore, it has been shown that proferrorosamine-negative Tn5 mutants of P. roseus fluorescens were able to form siderophores only under iron-limiting conditions when proferrorosamine A was added to the medium. Our data suggest that proferrorosamine A production is essential for siderophore synthesis by P. roseus fluorescens; the production of siderophores occurred only when proferrorosamine A was present.

Journal Article↗

Phosphonate utilization by bacteria in the presence of alternative phosphorus sources.

Batch and continuous culture experiments were carried out to investigate the effect of orthophosphate and p-nitrophenylphosphate on the utilization of various phosphonates as a P source by bacteria. Detailed tests with methylphosphonate as a model phosphonate and the phosphonate-degrading Pseudomonas paucimobilis strain MMM101a revealed that, in contrast with the majority of literature data, the phosphates did not suppress phosphonate utilization. Under conditions of P stress, strain MMM101a simultaneously took up both P-sources, with a preference for the phosphate-P. Study of the kinetic parameters for strain MMM101a, growing on the different P sources revealed similar, rather low, maximum growth rates (ca. 0.15 h-1). However, the affinity for orthophosphate (Ks:0.17 microM), was more than two orders of magnitude higher than for methylphosphonate (Ks: 66 microM), which might account for the preferential uptake of orthophosphate. Cellular phosphorus yields in continuous cultures varied considerably with the conditions applied. The results suggest that phosphonate degradation can occur also in environments with substantial backgrounds of phosphate.

Biodegradation, Environmental↗

Manganese as a controlling factor in mixed cultures of Lactobacillus plantarum and Enterobacteriaceae.

Addition of manganese, at levels of 50 ppm, to a liquid growth medium simulating adverse silage conditions had no effect on the growth or on the fermentation pattern of Enterobacter cloacae and Proteus vulgaris. Yet, the manganese strongly enhanced the growth of Lactobacillus plantarum. Co-cultures of L. plantarum and E. cloacae or P. vulgaris were, by addition of manganese ions, significantly altered in the favour of the former. This finding can be of use in mixed cultures where Enterobacteriaceae act as spoiler microorganisms.

Acetates↗

Iron complexation as a tool to direct mixed clostridia-lactobacilli fermentations.

Iron complexation was investigated as a possible tool to give lactobacilli a competitive advantage over clostridia. The iron complexing substance tested, i.e. 2,2'-dipyridyl, was not toxic itself for clostridia, but its addition to a mixed culture of lactobacilli and clostridia resulted in a strong ecological advantage of the lactobacilli.

2,2'-Dipyridyl↗

Marking the rhizopseudomonas strain 7NSK2 with a Mu d(lac) element for ecological studies.

The mini Mu element Mu dII1681, which contains the lac operon genes and a kanamycin resistance gene, was inserted in the chromosome of plant growth-beneficial Pseudomonas aeruginosa 7NSK2 to construct a marked strain (MPB1). In MPB1, beta-galactosidase is permanently expressed under the culture conditions used. The MPB1 strain could be recovered with an efficiency of about 100% from a sandy loam soil on 5-bromo-4-chloro-3-indolyl-beta-D-galactopyranoside medium containing sebacic acid and kanamycin. The limit of detection is about 10 CFU/g of soil. A detailed comparison was made between the wild-type strain 7NSK2 and the Mu dII1681-containing MPB1 strain. The results showed that no genes essential for growth, siderophore production, survival in sterile and nonsterile conditions, plant growth stimulation, or root colonization had been damaged in the MPB1 strain, which means that MPB1 can reliably be used for ecological studies in soil. MPB1 survived well at 4 or 28 degrees C but died off relatively rapidly in air-dried soil or at subzero temperatures. In these conditions, however, the MPB1 strain did not completely disappear from the soil but survived at a very low level of about 100 CFU/g of soil for more than 3 months. This observation stresses the need for very sensitive counting methods for ecological studies and for the evaluation of released microorganisms. Maize was inoculated with MPB1 via seed inoculation or soil inoculation. Upon seed inoculation, only the upper root parts were effectively colonized, while soil inoculation resulted in a complete colonization of the root system.

Ecology↗