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PubMed2006

Antagonistic coevolution with parasites increases the cost of host deleterious mutations.

The fitness consequences of deleterious mutations are sometimes greater when individuals are parasitized, hence parasites may result in the more rapid purging of deleterious mutations from host populations. The significance of host deleterious mutations when hosts and parasites antagonistically coevolve (reciprocal evolution of host resistance and parasite infectivity) has not previously been experimentally investigated. We addressed this by coevolving the bacterium Pseudomonas fluorescens and a parasitic bacteriophage in laboratory microcosms, using bacteria with high and low mutation loads. Directional coevolution between bacterial resistance and phage infectivity occurred in all populations. Bacterial population fitness, as measured by competition experiments with ancestral genotypes in the absence of phage, declined with time spent coevolving. However, this decline was significantly more rapid in bacteria with high mutation loads, suggesting the cost of bacterial resistance to phage was greater in the presence of deleterious mutations (synergistic epistasis). As such, resistance to phage was more costly to evolve in the presence of a high mutation load. Consistent with these data, bacteria with high mutation loads underwent less rapid directional coevolution with their phage populations, and showed lower levels of resistance to their coevolving phage populations. These data suggest that coevolution with parasites increases the rate at which deleterious mutations are purged from host populations.

Directed Molecular Evolution↗

PubMed2004

Adaptation of sympatric Achromatium spp. to different redox conditions as a mechanism for coexistence of functionally similar sulphur bacteria.

Changes in the abundance of sympatric Achromatium spp. in response to the artificial manipulation of redox conditions in sediment microcosms was determined by fluorescence in situ hybridization (FISH). Adaptation to different redox conditions was shown to be one mechanism that supported the coexistence of functionally similar Achromatium spp. In sediment microcosms, in which the overlying water was oxygenated, Achromatium community size and composition remained unchanged over time. However, imposition of anoxic conditions induced changes in community structure. Anoxia caused a reduction in the relative abundance of Achromatium sp. RY8 (72 +/- 4% to 49 +/- 2%) and an increase in Achromatium sp. RY5 (19 +/- 5% to 32 +/- 3%) and a newly identified Achromatium sp., RYKS (14 +/- 4% to 27 +/- 2%). In anoxic microcosms supplemented with a single addition of nitrate at different initial concentrations the relative decline in Achromatium sp. RY8 was dependent on the initial nitrate concentration. In these experiments nitrate was rapidly removed. In contrast, when high levels of nitrate were maintained by periodic replacement of the overlying water with nitrate supplemented anoxic water, the composition of the Achromatium community remained stable over time. This suggested that all of the coexisting Achromatium spp. are obligate or facultative anaerobes, but, Achromatium sp. RY8 was more sensitive to sediment redox conditions than the other Achromatium species. Given the heterogeneous nature of sedimentary environments, redox-related niche differentiation may promote coexistence of sympatric Achromatium spp.

Adaptation, Physiological↗

PubMed2000

Rhizosphere microbial community structure in relation to root location and plant iron nutritional status.

Root exudate composition and quantity vary in relation to plant nutritional status, but the impact of the differences on rhizosphere microbial communities is not known. To examine this question, we performed an experiment with barley (Hordeum vulgare) plants under iron-limiting and iron-sufficient growth conditions. Plants were grown in an iron-limiting soil in root box microcosms. One-half of the plants were treated with foliar iron every day to inhibit phytosiderophore production and to alter root exudate composition. After 30 days, the bacterial communities associated with different root zones, including the primary root tips, nonelongating secondary root tips, sites of lateral root emergence, and older roots distal from the tip, were characterized by using 16S ribosomal DNA (rDNA) fingerprints generated by PCR-denaturing gradient gel electrophoresis (DGGE). Our results showed that the microbial communities associated with the different root locations produced many common 16S rDNA bands but that the communities could be distinguished by using correspondence analysis. Approximately 40% of the variation between communities could be attributed to plant iron nutritional status. A sequence analysis of clones generated from a single 16S rDNA band obtained at all of the root locations revealed that there were taxonomically different species in the same band, suggesting that the resolving power of DGGE for characterization of community structure at the species level is limited. Our results suggest that the bacterial communities in the rhizosphere are substantially different in different root zones and that a rhizosphere community may be altered by changes in root exudate composition caused by changes in plant iron nutritional status.

Bacteria↗

PubMed2002

Carbon and hydrogen isotopic fractionation during biodegradation of methyl tert-butyl ether.

Carbon and hydrogen isotopic fractionation during aerobic biodegradation of MTBE by a bacterial pure culture (PM1) and a mixed consortia from Vandenberg Air Force Base (VAFB) were studied in order to assess the relative merits of stable carbon versus hydrogen isotopic analysis as an indicator of biodegradation. Carbon isotopic enrichment in residual MTBE of up to 8.1/1000 was observed at 99.7% biodegradation. Carbon fractionation was reproducible in the PM1 and VAFB experiments, yielding similar enrichment factors (epsilon) of -2.0/1000 +/- 0.1/1000 to -2.4/1000 +/- 0.3/1000 for replicates in the PM1 experiment and -1.5/1000 +/- 0.1/1000 to -1.8/1000 +/- 0.1/1000 for replicates in the VAFB experiment. Hydrogen isotopic fractionation was highly reproducible for the PM1 pure cultures, with epsilon values of -33/1000 +/- 5/1000 to -37/1000 +/- 4/1000 for replicate samples. In the VAFB microcosms, there was considerably more variability in epsilon values, with values of -29/1000 +/- 4/1000 and -66/1000 +/- 3/1000 measured for duplicate sample bottles. Despite this variability, hydrogen isotopic fractionation always resulted in 2H enrichment of the residual MTBE of >80/1000 at 90% biodegradation. The reproducible carbon fractionation suggests that compound-specific carbon isotope analysis may be used to estimate the extent of biodegradation at contaminated sites. Conversely, the large hydrogen isotopic fractionation documented during biodegradation of MTBE suggests that compound-specific hydrogen isotope analysis offers the most conclusive means of identifying in-situ biodegradation at contaminated sites.

Bacteria↗

PubMed1999

Effect of toxic metals on indigenous soil beta-subgroup proteobacterium ammonia oxidizer community structure and protection against toxicity by inoculated metal-resistant bacteria.

Contamination of soils with toxic metals is a major problem on military, industrial, and mining sites worldwide. Of particular interest to the field of bioremediation is the selection of biological markers for the end point of remediation. In this microcosm study, we focus on the effect of addition of a mixture of toxic metals (cadmium, cobalt, cesium, and strontium as chlorides) to soil on the population structure and size of the ammonia oxidizers that are members of the beta subgroup of the Proteobacteria (beta-subgroup ammonia oxidizers). In a parallel experiment, the soils were also treated by the addition of five strains of metal-resistant heterotrophic bacteria. Effects on nitrogen cycling were measured by monitoring the NH3 and NH4+ levels in soil samples. The gene encoding the alpha-subunit of ammonia monooxygenase (amoA) was selected as a functional molecular marker for the beta-subgroup ammonia oxidizing bacteria. Community structure comparisons were performed with clone libraries of PCR-amplified fragments of amoA recovered from contaminated and control microcosms for 8 weeks. Analysis was performed by restriction digestion and sequence comparison. The abundance of ammonia oxidizers in these microcosms was also monitored by competitive PCR. All amoA gene fragments recovered grouped with sequences derived from cultured Nitrosospira. These comprised four novel sequence clusters and a single unique clone. Specific changes in the community structure of beta-subgroup ammonia oxidizers were associated with the addition of metals. These changes were not seen in the presence of the inoculated metal-resistant bacteria. Neither treatment significantly altered the total number of beta-subgroup ammonia-oxidizing cells per gram of soil compared to untreated controls. Following an initial decrease in concentration, ammonia began to accumulate in metal-treated soils toward the end of the experiment.

Ammonia↗

PubMed2006

[The time-evolution study on the laser-induced oxygen plasmas spectra].

The present paper reports the results of spectroscopic study on the oxygen plasma induced by a pulsed laser with a wavelength of 1.06 microm. The gas samples were pure oxygen (99.9999%) and the spectral range was from 300 to 900 nm. The temporal behavior of the spectra was measured. The experiment results showed that the spectra could be divided into continuous spectra and line spectra. Each component of the spectra attenuated according to the disparate mode along temporal development. The microcosmic physical processes and mechanisms were analyzed. The reason for the longer plasma spectral lifetime was discussed. These works were helpful to better understanding of the microscopic mechanisms during the decay of the laser-induced air plasmas, and also provide more experimental evidences for prolonging the decay time of plasmas in some technical applications.

English Abstract↗

PubMed2003

The effect of amount of crude oil on extent of its biodegradation in open water- and sandy beach-laboratory simulations.

We examined the biodegradation of varying amounts of artificially weathered Alaskan North Slope crude oil in laboratory microcosm test systems that use natural seawater and simulate spills in open water and on sandy beaches. The model bioremediation treatment consisted of periodic applications of marine bacteria, selected to degrade n-alkanes and a range of aromatic compounds, suspended in a salts solution that supplied inorganic nitrogen and phosphorous. Beach microcosms dosed with low and high oiling lost an average of 22.5% and 11.3% oil weight, respectively. Open-water microcosms dosed with high and low oiling lost 19.1% and 2.9% oil weight, respectively. Thus, the lower doses of oil were more efficiently degraded. The model bioremediation treatment also affected a greater number of selected analytical endpoints in the lower-oil-dose than higher-dose experiments and the former showed more substantial degradation of recalcitrant components. Above a certain threshold oil concentration, bioremediation did not effectively remove oil. Below this threshold the distinction between active bioremediation treatment and intrinsic biodegradation of the controls was less prominent; i.e., fewer of the oil components were statistically depleted by remediation treatment relative to controls. Furthermore, the oil-dose range over which bioremediation was realized in these systems occurred at very low oiling levels. Thus, under the environmental conditions simulated in these microcosms, the effectiveness of bioremediation peaked over a rather narrow low-dose oiling range.

Biodegradation, Environmental↗

PubMed2005

Effect of dissolved oxygen regime on growth dynamics of Pseudomonas spp during benzene degradation.

We investigated the effect of different oxygen regimes on growth patterns of Pseudomonas spp. during benzene degradation in microcosm batch studies. Benzene degradation was induced by limiting oxygen available for microbial activity, which consists of three initial-dissolved oxygen (DO) levels of oxic, hypoxic, and anoxic conditions. Batch experiments were performed for cell growth and benzene degradation by inoculating three strains of Pseudomonas spp. (Pseudomonas aeruginosa, Pseudomonas fluorescens, Pseudomonas putida) in mineral salt medium containing aqueous benzene. Results showed that all strains were capable to grow and degrade benzene under all oxygen regimes but in a different manner. The highest cell growth of P. aeruginosa and P. fluorescens was achieved under oxic and anoxic condition, respectively, but there was no substantial difference on benzene degradation between the oxygen treatments with about 25% reduction for both strains. P. putida showed a facultative process for both cell growth and benzene degradation. This reveals that care should be taken in selection of microorganisms with regard to environmental studies since they exhibit different responses for given environmental conditions such as DO levels.

Benzene↗

PubMed2006

Crude petroleum-oil biodegradation efficiency of Bacillus subtilis and Pseudomonas aeruginosa strains isolated from a petroleum-oil contaminated soil from North-East India.

The efficiency of Bacillus subtilis DM-04 and Pseudomonas aeruginosa M and NM strains isolated from a petroleum contaminated soil sample from North-East India was compared for the biodegradation of crude petroleum-oil hydrocarbons in soil and shake flask study. These bacterial strains could utilize crude petroleum-oil hydrocarbons as sole source of carbon and energy. Bioaugmentation of TPH contaminated microcosm with P. aeruginosa M and NM consortia and B. subtilis strain showed a significant reduction of TPH levels in treated soil as compared to control soil at the end of experiment (120 d). P. aeruginosa strains were more efficient than B. subtilis strain in reducing the TPH content from the medium. The plate count technique indicated expressive growth and biosurfactant production by exogenously seeded bacteria in crude petroleum-oil rich soil. The results showed that B. subtilis DM-04 and P. aeruginosa M and NM strains could be effective for in situ bioremediation.

Bacillus subtilis↗

PubMed2005

Does sediment grain size affect diatom grazing by harpacticoid copepods?

Estuarine soft sediments support a diverse group of eukaryotic and prokaryotic organisms though the role of the sediment per se for the functioning of these organisms remains largely unknown. The present study aimed to test the effect of sediment grain size on the grazing activities of harpacticoid copepods. In controlled experiments, two common intertidal harpacticoid species (Paramphiascella fulvofasciata and Nitokra spinipes) were each offered a mix of two benthic diatom species (Navicula phyllepta and Seminavis robusta) in different sedimentary conditions. Several microcosms were created using a variety of sediment types, including fine silt (<63 microm), coarser grained sands (125-250, 250-450, 100-300 microm), artificial 'sediments' of glass beads (250-500, 2000 microm) and even the absence of sediment was tested. The diatoms were enriched in the stable carbon (13)C to facilitate tracing in the harpacticoids. Both copepod species were able to graze on the diatoms with highest uptake when sediment was absent. In contrast, both harpacticoid species showed no uptake in silty conditions. In general, grazing was favoured when mean sediment grain size increased. The strong negative effect of fine grains on the grazer's efficiency can be explained by the resulting differences in the structure (and accessibility) of the diatom biofilm on the one hand and the mobility of the grazer on the other hand. In view of the subtle equilibrium between primary producers and grazers, these results might have important implications for the effect of siltation of tidal flats due to, e.g., human activities.

Analysis of Variance↗

PubMed2004

Biodiversity effects on soil processes explained by interspecific functional dissimilarity.

The loss of biodiversity can have significant impacts on ecosystem functioning, but the mechanisms involved lack empirical confirmation. Using soil microcosms, we show experimentally that functional dissimilarity among detritivorous species, not species number, drives community compositional effects on leaf litter mass loss and soil respiration, two key soil ecosystem processes. These experiments confirm theoretical predictions that biodiversity effects on ecosystem functioning can be predicted by the degree of functional differences among species.

Animals↗

PubMed2006

Dispersal of Beauveria bassiana by the activity of nettle insects.

Recent studies have shown that the entomopathogenic fungus Beauveria bassiana occurs naturally on the phylloplanes of several plants, including nettles. Insects could, by their activity, be contributing to this inoculum by dispersing it from other sites. The potential of nettle aphids Microlophium carnosum and their predator Anthocoris nemorum to disperse conidia of B. bassiana from soil to nettles and from sporulating cadavers in the nettle canopy was investigated in laboratory experiments. In petri dish assays, aphids showed potential to distribute B. bassiana from soil to nettle leaves. Predators dispersed inoculum from both soil and cadavers to nettle leaves in petri dishes. In microcosms, aphids did not disperse B. bassiana from the soil or from cadavers confined in the canopy, but A. nemorum were able to transfer inoculum from soil into the nettle canopy and to distribute conidia from cryptic cadavers. In some instances, infections were initiated in aphids and predators as a consequence of dispersal.

Animals↗

PubMed1994

Evidence of a dormant but infective state of the fish pathogen Pasteurella piscicida in seawater and sediment.

The stability of Pasteurella piscicida strains in seawater and sediment microcosms at different temperatures (6 and 20 degrees C) was investigated during a 1-month period. Three strains of P. piscicida showed similar survival kinetics. By a standard plate count method they survived in water and sediment for only 6 to 12 days, depending on the strain and type of microcosm. During this starvation period, the metabolic activity of the cells was reduced by more than 80%. Culturable cells of each P. piscicida strain persisted better in sediment than in water, as well as at 20 degrees C compared to 6 degrees C. However, in all the microcosms, the acridine orange direct counts remained at about 10(5) cells per ml during the experimental period, which demonstrated that P. piscicida possesses a capacity to enter a viable but not culturable state. Moreover, dormant cells were always resuscitated by the addition of fresh medium to the microcosms, since we recovered numbers of culturable cells similar to the acridine orange direct counts. These resuscitated cells exhibited the same respiration rate as that seen prior to the start of the experiments. Although the biochemical, physiological, and serological characteristics; lipopolysaccharides; membrane proteins; and plasmid content of P. piscicida strains were unaffected during the starvation conditions, the dormant cells were smaller (dwarf cells) and had increased surface hydrophobicity. The starved cells maintained their infectivity and pathogenic potential for fish, with 50% lethal doses similar to those of the original strains.

Animals↗

PubMed2003

Effect of sorption on benzene biodegradation in sandy soil.

The effect of sorption on benzene biodegradation in sandy soil was studied by conducting kinetic microcosm batch tests in soil-free solution and in the presence or absence of bacteria in soil materials with varying degrees of powdered activated carbon (PAC). In the soil-free experiment, benzene was added to a solution inoculated with Pseudomonas aeruginosa bacteria in order to achieve a potential or maximum biodegradation rate. In subsequent experiments, benzene was applied to a solution containing sandy soil and various PAC contents with and without inoculating P. aeruginosa. Benzene concentrations in the soil-free experiments decreased with time with two characteristic rates. A two-stage exponential decay model adequately represented the observed solution concentration pattern with time. Sorption experiments in bacteria-free soil also decreased monotonically, with the extent of sorption increasing as PAC content increased. The sorption data were represented well with a two-stage irreversible sorption model. A third set of experiments in the presence of both soil and bacteria showed more rapid concentration loss from solution than the set of experiments with bacteria-free soil. A model combining sorption and degradation greatly overestimated the loss when the rate coefficient from the bacteria-free experiments was used. Satisfactory agreement between model predictions and observed values was obtained when the degradation rate coefficients were decreased by factors ranging from 3 to 10, depending on the amount of PAC present. Model predictions of the percentage benzene mass remaining in the soil after 25 d of degradation ranged from 72 to 97%, depending on the PAC content, compared to only 2.5% remaining in soil-free solution.

Adsorption↗

PubMed1992

Expression and transfer of engineered catabolic pathways harbored by Pseudomonas spp. introduced into activated sludge microcosms.

Two genetically engineered microorganisms (GEMs), Pseudomonas sp. strain B13 FR1(pFRC20P) (FR120) and Pseudomonas putida KT2440(pWWO-EB62) (EB62), were introduced into activated sludge microcosms that had the level of aeration, nutrient makeup, and microbial community structure of activated sludge reactors. FR120 contains an experimentally assembled ortho cleavage route for simultaneous degradation of 3-chlorobenzoate (3CB) and 4-methyl benzoate (4MB); EB62 contains a derivative TOL plasmid-encoded degradative pathway for toluene experimentally evolved so that it additionally processes 4-ethyl benzoate (4EB). Experiments assessed survival of the GEMs, their ability to degrade target substrates, and lateral transfer of plasmid-encoded recombinant DNA. GEMs added at initial densities of 10(6) to 10(7) bacteria per ml of activated sludge declined to stable population densities of 10(4) to 10(5) bacteria per ml. FR120 degraded combinations of 3CB and 4MB (1 mM each) following 3 days of adaptation in the microcosms. Indigenous microorganisms required an 8-day adaptation period before degradation of 4MB was observed; 3CB was degraded only after the concentration of 4MB was much reduced. The indigenous microbial community was killed when both compounds were present at concentrations of 4.0 mM. However, in parallel microcosms containing FR120, the microbial community maintained a normal density of viable cells. Indigenous microbes readily degraded 4EB (2 mM), and EB62 did not significantly increase the observed rate of degradation. In filter matings, transfer of pFRC20P, which specifies mobilization but not transfer functions, from FR120 to P. putida UWC1 was not detectable (< 10(-7) transconjugants per donor cell).(ABSTRACT TRUNCATED AT 250 WORDS)

Benzoates↗

PubMed2004

Learning and living health: college students' experiences with an introductory health course.

Despite the Healthy People 2010 initiative and the national focus on promoting healthy lifestyle strategies, studies indicate that U.S. citizens are becoming more obese, and that rates of depression, diabetes, hypertension, homicide, and suicide are increasing. The college campus provides a microcosm of the health issues facing the nation, with college students' reports of increased risk behaviors evident in higher rates of smoking and binge drinking. Faculty in schools of nursing are in an ideal position to develop and provide health promotion and illness prevention programs, both for their own nursing students and for a broader campus-wide student population. This article describes a research study that course faculty conducted during the first year of implementation, designed to evaluate the extent to which participation in the "Introduction to Health" coursework and related activities influenced students' health-related behaviors.

Adaptation, Psychological↗

PubMed2002

Ozone effects on Sphagnum mosses, carbon dioxide exchange and methane emission in boreal peatland microcosms.

Microcosms of a boreal peatland originating from an oligotrophic fen in Eastern Finland were fumigated under four ozone concentrations (0, 50, 100 and 150 ppb O3) in laboratory growth chambers during two separate experiments (autumn and summer) for 4 and 6 weeks, respectively. Ozone effects on Sphagnum mosses and the fluxes of carbon dioxide and methane were evaluated. In both experiments, the three Sphagnum species studied showed only a few significant responses to ozone. In the autumn experiment, membrane permeability of S. angustifolium, measured as conductivity and magnesium leakage, was significantly higher under ozone fumigation (P = 0.005 and < 0.001, respectively), and there was a distinct dose-dependence. S. magellanicum showed no clear responses, either for membrane leakage or pigment content. There were no substantial ozone responses in the gross photosynthesis or net CO2 exchange during the 6-week-long summer experiment, but dark ecosystem respiration was transiently increased by ozone concentration of 100 ppb after 14 days of exposure (P < 0.05). Fumigation with 100 ppb of ozone, however, more than doubled (P < 0.05) methane emission from the peatland monoliths. Our results suggest that increasing tropospheric ozone concentration may cause substantial changes in the carbon gas cycling of boreal peatlands, even though these changes are not closely associated with the changes in Sphagnum vegetation.

Bryopsida↗

PubMed2002

In situ MTBE biodegradation supported by diffusive oxygen release.

Microcosm studies with sediments from Vandenberg Air Force Base, CA, suggest that native aerobic methyl tert-butyl ether (MTBE)-degrading microorganisms can be stimulated to degrade MTBE. In a series of field experiments, dissolved oxygen has been released into the anaerobic MTBE plume by diffusion through the walls of oxygen-pressurized polymeric tubing placed in contact with the flowing groundwater. MTBE concentrations were decreased from several hundred to less than 10 microg/L during passage through the induced aerobic zone, due apparently to in situ biodegradation: abiotic MTBE loss mechanisms were insignificant. Lag time for initiation of degradation was less than 2 months, and the apparent pseudo-first-order degradation rate was 5.3 day(-1). Additional MTBE was added in steps to raise the influent concentration to a maximum of 2.1 mg/L. With each step, MTBE was degraded within the preestablished aerobic treatment zone at rates ranging from 4.4 to 8.6 day(-1). Excess dissolved oxygen suggested that even higher MTBE concentrations could have been treated. Continued flow through the treatment zone was repeatedly confirmed through tracer and other tests. These and others' results suggest that it is possible to create permeable in situ treatment zones solely by releasing oxygen to support native microbial degradation of MTBE.

Bacteria, Anaerobic↗