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PubMed1979

Sampling culturable heterotrophs from microcosms: a statistical analysis.

The contributions of different sources of error in sampling mixed and unmixed bacterial microcosms were evaluated by using analysis of variance. Culturable heterotrophic bacteria from a turbid freshwater impoundment were sampled from 9-liter tanks that were unagitated or mixed with magnetic stirrers or pumps and from dilution bottles that were unagitated or agitated with a mechanical shaker. Axenic cultures of Enterobacter aerogenes were also sampled from manually shaken test tubes. In both agitated and unagitated tanks and in unagitated dilution bottles, dilutions made from the same sampling pipette were significantly different, showing a clumping of bacteria on the scale of millimeters. Also, microcosms within a single experiment differed from one another by a large margin. Dilution mean squares and tank or bottle mean squares were homogeneous for all types of tanks and unagitated bottles, indicating that the gentle mixing provided by pumps and stir bars did not reduce either millimeter scale or intermicrocosm variability over what prevailed in unagitated microcosms. By contrast, the vigorously shaken bottles and test tubes showed no millimeter scale variability. Intermicrocosm variability was undetectable in test tubes and two orders of magnitude less in shaken bottles than in unshaken bottles. When these facts are coupled with the inherent statistical advantage of replicating large rather than small experimental units, it is concluded that sampling error in the enumeration of aquatic bacteria in microcosms will be reduced by using numerous, small, violently agitated microcosms with a minimum of subsampling per microcosm.

Analysis of Variance↗
PubMed2003

Moisture conditions and the presence of bryophytes determine fescue species abundance in a dry calcareous grassland.

Festuca ovina is the abundant matrix-forming species and F. rubra a subordinate species in shallow-soil calcareous grasslands. F. pratensis is a transient species, occurring sparsely in this community. We hypothesised that the different abundances of these three species are primarily due to the differential effect of moisture conditions on their germination and early establishment, and that the effect of the pattern of rainfall intensity depends on the presence or absence of a bryophyte layer. We studied the dependence of the germination and establishment of the three fescue species on the moisture conditions both in the laboratory and in the patches of intact grassland community (microcosms). In a laboratory germination experiment, F. pratensis showed the highest, F. rubra, the intermediate and F. ovina, the lowest drought tolerance. In microcosms, the establishment of F. ovina was the highest. At the same time, the annual mortality of seedlings of F. ovina was the lowest. All three species responded positively to an increasing irrigation level. Differently from F. ovina, F. rubra showed a positive response only in plots from which the bryophyte layer had been removed, while F. pratensis responded positively to both irrigation and bryophyte removal. We conclude that moisture conditions have a differential effect on the three fescue species mainly in the seedling establishment, not in the germination phase. For the successful establishment of F. rubra and F. pratensis, the coincidence of high rainfall and local disturbance, removing bryophytes, is required. The presence or absence of bryophytes had no effect on establishment in dry years, while in rainy years the removal of bryophytes has a clear positive effect.

Bryophyta↗
PubMed1994

Behavioral and physiological responses of the terrestrial life stages of the Jefferson salamander, Ambystoma jeffersonianum, to low soil pH.

Terrestrial metamorphs of the salamander, Ambystoma jeffersonianum, were exposed to four levels of pH on artificial substrates in the laboratory. Short-term exposure (7 d) to pH levels between 3.5 and 5.0 had no effect on total body mass, Na+, K+, Ca2+, and Mg2+ concentrations. However, at 14 d in both laboratory experiments and in field microcosms, whole body water and Na+ concentrations were significantly lower at the lower pH level. Terrestrial metamorphs and adults of the Jefferson salamander preferred artificial substrates that were higher in pH. Ponds not utilized for breeding all had adjacent soil pH below 3.7. Pond water and soil pH were directly and highly correlated. Soil pH may indirectly affect breeding success of Jefferson salamanders by influencing pond pH or directly by physiologically stressing the terrestrial life stages. Laboratory data implicate direct effects on the physiology of terrestrial young of the year salamanders by induction of loss of body water and Na+. Apparently, soil pH is a critical factor not only in the selection of vernal ponds by Jefferson salamander adults for breeding, but in determining the likelihood of reproductive success as judged by recruitment into the adult population.

Ambystoma↗
PubMed2002

Survival and leaching of fecal bacteria after slurry spreading on mountain hay meadows: consequences for the management of water contamination risk.

The aim of this work is to achieve a better understanding of the behavior of fecal coliform populations on the vegetation and in the soil after slurry spreading in environmental conditions, typical of vegetative growth period in mountain hay meadows. Changes in fecal coliform populations on the vegetation and in the soil were monitored in situ for 3 months after slurry spreading on 9 plots. The variations found in populations are related to the agricultural, soil. and climatic characteristics of plots and to the moisture regime of soils. These observations are compared with laboratory experiments on undisturbed soil microcosms. In absence of water flux, survival durations recorded in the laboratory and in the field are of the same order of magnitude. The data enable us to pinpoint the influence of various factors affecting the decline and transfer of fecal bacteria in the plant-soil system and consequently to discuss the risk management of water contamination by agriculture.

Agriculture↗
PubMed1998

Degradation of 2,4,6-trichlorophenol by a specialized organism and by indigenous soil microflora: bioaugmentation and self-remediability for soil restoration.

A selected mixed culture and a strain of Alcaligenes eutrophus TCP were able to totally degrade 2,4,6,-TCP with stoichiometric release of Cl-. In cultures of Alc. eutrophus TCP, a dioxygenated dichlorinated metabolite was detected after 48 h of incubation. Experiments conducted with soil microcosms gave evidence that: the degradative process had a biotic nature and was accompanied by microbial growth; the soil used presented an intrinsic degradative capacity versus 2,4,6-TCP; the specialized organism used as inoculum was effective in degrading 2,4,6-TCP in a short time. These results could be utilized for the adoption of appropriate remediation techniques for contaminated soil.

Alcaligenes↗
PubMed2001

Combined effects of predatory fish and sublethal pesticide contamination on the behavior and mortality of mayfly nymphs.

We evaluated the potential interaction of pesticide effect and predatory fish on behavior and mortality of a stream mayfly. Experiments in laboratory stream microcosms compared the activity, drift, and mortality of Baetis mayfly nymphs in the absence of fish with that in the presence of Cape galaxias (Galaxias zebratus), both species inhabiting the same stream environments in the Western Cape of South Africa. These two predator treatments were combined with exposure either to no pesticide or to 0.2 microg/L of the organophosphate insecticide azinphos-methyl (AZP) or 0.2 microg/L of the pyrethroid insecticide fenvalerate (FV). Such pesticide levels are known from transient spraydrift- or runoff-related pesticide input into running waters. Each of the six combinations was replicated six times as 30-min trials during the day and effects were analyzed using 2 x 2 factorial analysis of variance (ANOVA). A single exposure to either fish or pesticide significantly increased the absolute activity of mayflies, measured as number of animals visible on top of stones, and the absolute mayfly drift in the fish treatment and in the FV treatment but did not increase the mortality above 0.8%. The combination of predator presence and sublethal pesticide exposure resulted in a significant increase in the mortality rate, to about 9% in the AZP x fish and 25% in the FV x fish treatment, although the activity and drift rates were not increased compared with the single-stressor treatments. Two-by-two factorial ANOVA and the comparison of expected and measured responses indicated that the mortality resulted from a synergistic interaction of the two stressors. The observed mortality was without exception caused by predation of the fish on drifting mayflies. The relative drift rate in the FV x fish treatment was decreased, again due to a synergistic interaction, which suggests an active drift avoidance reaction of the mayflies exposed to the combined pesticide x fish treatment, in contrast with those exposed just to FV. We conclude that the drift response of mayflies to transient sublethal pesticide exposure results in a synergistically increased adverse effect in the presence of predatory fish.

Animals↗
PubMedDate not supplied

Fate and toxicity of aircraft deicing fluid additives through anaerobic digestion.

Benzotriazole derivatives are widely used corrosion inhibitors and their fate during wastewater treatment processes is unknown. The purpose of this research was to study the toxic effects and fate of the two commercially significant benzotriazole isomers used in aircraft deicing fluids (4-, and 5-, methylbenzotriazole [MeBT]) during anaerobic digestion. Experiments were executed in microcosms using mesophilic anaerobic biomass co-digesting wastewater sludge and propylene glycol. Sorption of MeBT to digesting solids could be approximated with a Freundlich model, and no anaerobic breakdown of either MeBT isomer was detected. Digesters fed more than 300 mg/L MeBT responded with a significant decrease in methanogenic microbial activity and volatile solids production and a concomitant increase in accumulation of volatile fatty acids. Direct microscopic measurements using fluorescent phylogenetic probes applied to digesting biomass revealed that members of both Archaea and Bacteria domains were sensitive to MeBT. Granular activated carbon (GAC) (volatile solids: GAC = 10%) reduced the apparent toxic effects of MeBT; GAC addition nearly restored the baseline activity of digesters fed MeBT (500 to 1000 mg/L).

Aircraft↗
PubMed2005

Using phospholipid fatty acid technique to study short-term effects of the biological control agent Pseudomonas fluorescens DR54 on the microbial microbiota in barley rhizosphere.

The biological control agent (BCA) Pseudomonas fluorescens DR54 was applied to seeds (experiment 1) or roots (experiment 2) of barley growing in microcosms, while noninoculated plants served as controls. The fate of the BCA and its effects on the rhizosphere microbial community was evaluated in microcosms destructively sampled at days 2, 4, 6, and 9 after inoculation. In both experiments the number of P. fluorescens DR54 cells decreased immediately after application as enumerated by immunostaining and microscope direct counting. Substrate-induced respiration (SIR) was taken as a measurement of the active microbial biomass, while indicators of the total microbiota (and main taxonomic groups) were obtained using the phospholipid fatty acid (PLFA) technique. In experiment 1, these parameters were unaffected by the relatively small number of BCA cells applied, whereas in experiment 2, the larger BCA input resulted in an enhanced level of both SIR and PLFAs from Gram-negative bacteria (which included the BCA itself). However, at day 9 after inoculation, treatments with P. fluorescens DR54 and controls were similar in all measured parameters in both experiments. This was also illustrated very clearly by principal component analysis of the PLFA data, which in both experiments were able to discriminate between treatments in the first days after BCA inoculation, thus confirming the sensitivity of this method. Laccase activity has a potential as an indicator of fungal stress, e.g., when challenged with an antifungal BCA. This seemed to be supported in experiment 2, where the activity of this enzyme was enhanced four-fold in the BCA treatment at day 2. Our study shows that under the present conditions, P. fluorescens DR54 disappears from the soil and causes only transient effects on the soil microbiota. It also shows that the PLFA technique is a sensitive and reliable monitoring tool in in situ assessment of BCA nontarget effect on indigenous microorganisms in soil.

Fatty Acids↗
PubMed2002

Robust hydrocarbon degradation and dynamics of bacterial communities during nutrient-enhanced oil spill bioremediation.

Degradation of oil on beaches is, in general, limited by the supply of inorganic nutrients. In order to obtain a more systematic understanding of the effects of nutrient addition on oil spill bioremediation, beach sediment microcosms contaminated with oil were treated with different levels of inorganic nutrients. Oil biodegradation was assessed respirometrically and on the basis of changes in oil composition. Bacterial communities were compared by numerical analysis of denaturing gradient gel electrophoresis (DGGE) profiles of PCR-amplified 16S rRNA genes and cloning and sequencing of PCR-amplified 16S rRNA genes. Nutrient amendment over a wide range of concentrations significantly improved oil degradation, confirming that N and P limited degradation over the concentration range tested. However, the extent and rate of oil degradation were similar for all microcosms, indicating that, in this experiment, it was the addition of inorganic nutrients rather than the precise amount that was most important operationally. Very different microbial communities were selected in all of the microcosms. Similarities between DGGE profiles of replicate samples from a single microcosm were high (95% +/- 5%), but similarities between DGGE profiles from replicate microcosms receiving the same level of inorganic nutrients (68% +/- 5%) were not significantly higher than those between microcosms subjected to different nutrient amendments (63% +/- 7%). Therefore, it is apparent that the different communities selected cannot be attributed to the level of inorganic nutrients present in different microcosms. Bioremediation treatments dramatically reduced the diversity of the bacterial community. The decrease in diversity could be accounted for by a strong selection for bacteria belonging to the alkane-degrading Alcanivorax/Fundibacter group. On the basis of Shannon-Weaver indices, rapid recovery of the bacterial community diversity to preoiling levels of diversity occurred. However, although the overall diversity was similar, there were considerable qualitative differences in the community structure before and after the bioremediation treatments.

Bacteria↗
PubMed2005

Adaptive radiation in microbial microcosms.

It has often been argued that evolutionary diversification is the result of divergent natural selection for specialization on alternative resources. I provide a comprehensive review of experiments that examine the ecology and genetics of resource specialization and adaptive radiation in microbial microcosms. In these experiments, resource heterogeneity generates divergent selection for specialization on alternative resources. At a molecular level, the evolution of specialization is generally attributable to mutations that de-regulate the expression of existing biosynthetic and catabolic pathways. Trade-offs are associated with the evolution of resource specialization, but these trade-offs are often not the result of antagonistic pleiotropy. Replicate adaptive radiations result in the evolution of a similar assemblage of specialists, but the genetic basis of specialization differs in replicate radiations. The implications of microbial selection experiments for evolutionary theory are discussed and future directions of research are proposed.

Adaptation, Biological↗
PubMed2007

Atrazine sorption and fate in a Ultisol from humid tropical Brazil.

This study combined laboratory based microcosm systems as well as field experiments to evaluate the mobility of atrazine on a Ultisol under humid tropical conditions in Brazil. Results from sorption experiments fit to the Freundlich isotherm model [K(f) 0.99 mg kg(-1)/(mg l(-1))(1/n)], and indicate a low sorption capacity for atrazine in this soil and consequently large potential for movement by leaching and runoff. Microcosm systems using (14)C-atrazine to trace the fate of the applied herbicide, showed that 0.33% of the atrazine was volatilized, 0.25% mineralized and 6.89% was recorded in the leachate. After 60 d in the microcosms, 75% of the (14)C remained in the upper 5 cm soil layer indicating atrazine or its metabolites remained close to the soil surface. In field experiments, after 60 d, only 5% of the atrazine applied was recovered in the upper soil layers. In the field experiments atrazine was detected at a depth of 50 cm indicating leaching. Simulating tropical rain in field experiments resulted in 2.1% loss of atrazine in runoff of which 0.5% was adsorbed onto transported soil particles and 1.6% was in solution. Atrazine runoff was greatest two days after herbicide application and decreased 10 fold after 15 d. The use of atrazine on Ultisols, in the humid tropics, constitutes a threat to water quality, causing surface water and ground water pollution.

Adsorption↗
PubMed2005

Sediment resuspension effects on the benthic microbial loop in experimental microcosms.

Sediment resuspension induced by anthropogenic disturbance is becoming a major threat of marine coastal ecosystems worldwide. The effects of sediment resuspension on the pelagic domain and on macro- and meiobenthos are well documented in the literature, whereas the effects on the benthic microbial components are nearly neglected. We have investigated the effects of sediment resuspension at two different disturbance levels on benthic bacterial abundance, biomass, and activities and on heterotrophic nanobenthos abundance in experimental microcosms. The results of our experiments pointed out that, independently from the amount of involved energy, sediment resuspension determined a general decrease of all benthic microbial components. The main effects consisted of a decreased abundance of the metabolically active bacterial fraction and of heterotrophic nanobenthos abundance. However, the amount of energy involved in sediment resuspension had differential effects on the structure and functioning of the benthic microbial loop, but only in the short term (i.e., within 36 h). Sediment resuspension had a stimulatory effect on activities of surviving bacterial cells, which, in turn, resulted in increased sediment organic C turnover rates. We hypothesize that such an effect, enhancing nutrient availability, might have relevant consequences on the trophic state of coastal marine ecosystem.

Aminopeptidases↗
PubMed2001

Aspects of Marker/Reporter Stability and Selectivity in Soil Microbiology.

Based on several experiences of microbial release using genetically modified Rhizobium leguminosarum, we have highlighted a number of aspects related to the suitability of introduced markers such as resistance to mercury and b-galactosidase activity, the latter serving the function of high-expression level reporter gene obtained by the introduction of a synthetic promoter conferring strong inducible expression in Gram-negative bacteria. In vitro expression and in vivo performances of the chosen examples have been followed in model strains comparing gene dosage and expression levels. The technical possibility of unambiguously monitoring the marked GMM has been evaluated in medium- and long-term experiments carried out both in microcosms and soil, also including the presence of the plant symbiotic host. Marker stability, regardless the nature of the gene, was shown to be dependent on the location of the genetic modification and on its degree of gene expression regulation. Reporter strength was found to be an advantage allowing the distinction of marker-bearing bacteria while negatively affecting their genetic stability. Plasmid-borne regulated reporters were found to be stable up to the stages of rhizosphere colonization, but were more critically selected against upon symbiotic host invasion.

Journal Article↗
PubMed2003

A field method using microcosms to evaluate transfer of Cd, Cu, Ni, Pb and Zn from sewage sludge amended forest soils to Helix aspersa snails.

Juvenile Helix aspersa snails exposed in field microcosms were used to assess the transfer of Cd, Cu, Ni, Pb and Zn from forest soils amended with liquid and composted sewage sludge. Zn concentrations and contents were significantly higher in snails exposed to liquid and composted sludge after 5 and 7 weeks of exposure, when compared with control. Trends were less clear for the other metals. Present results show that Zn, among the cocktail of metallic trace elements (MTE) coming from sewage sludge disposal, represents the principal concern for food chain transfer and secondary poisoning risks. The microcosm design used in this experiment was well suited for relatively long-term (about 2 months) active biomonitoring with H. aspersa snails. The snails quickly indicated the variations of MTE concentrations in their immediate environment. Therefore, the present study provides a simple but efficient field tool to evaluate MTE bioavailability and transfer.

Animals↗
PubMed2005

Effect of manure on glyphosate and trifluralin mineralization in soil.

Manure additions to soil may alter soil chemical, physical, and biological characteristics, and thereby change pesticide fate processes in soil. This is the first study to examine the impact of liquid hog manure amendments on glyphosate and trifluralin mineralization in soil. Experiments were conducted in soil microcosms in the laboratory for a total of 332 (glyphosate) and 430 (trifluralin) days. The rate and amount of mineralization of both glyphosate and trifluralin were significantly influenced by the additions of fresh manure to soil in the laboratory and by the history of manure applications in the field. However, the maximum difference in herbicide mineralization between soils that were free of manure application and those amended with manure in the field or in the laboratory was only 6.1% and 7.3% of that initially applied, for trifluralin and glyphosate, respectively. Therefore, we conclude that liquid hog manure application to soil will have no significant effect on the mineralization of glyphosate and trifluralin under field conditions.

Animals↗
PubMed2002

Simulating species loss following perturbation: assessing the effects on process rates.

We removed stream-living macroinvertebrate shredder species in the sequences in which they are predicted to disappear, in response to two common types of anthropogenic disturbances: acidification and organic pollution, and analysed the effects on leaf breakdown rates. The experiment was performed in field microcosms using three shredder species. Species identity significantly affected leaf breakdown rates, while species richness per se was non-significant. The simulated sequential species loss showed large effects on leaf breakdown rates, with observed rates being significantly higher than expected from single-species treatments in two, out of four, two-species, and in all four three-species treatments. The invertebrates used in this study were taxonomically distinct (Insecta: Plecoptera and Trichoptera; Crustacea: Amphipoda), and of different sizes, hence a high degree of complementarity was probably present. A method to study the effects of species loss, characteristic of perturbation type, could be more useful than a random approach when investigating the impact of perturbation. Our results may have general applicability for investigations on the effects of diversity loss on ecosystem functioning in any ecosystem exposed to human perturbations, given that the order of extinction is known or can easily be assessed.

Amphipoda↗
PubMedDate not supplied

Microbial reduction of hexavalent chromium under vadose zone conditions.

Hexavalent chromium [Cr(VI)] is a common contaminant associated with nuclear reactors and fuel processing. Improper disposal at facilities in and and semiarid regions has contaminated underlying vadose zones and aquifers. The objectives of this study were to assess the potential for immobilizing Cr(VI) using a native microbial community to reduce soluble Cr(VI) to insoluble Cr(III) under conditions similar to those in the vadose zone, and to evaluate the potential for enhancing biological Cr(VI) reduction through nutrient addition. Batch microcosm and unsaturated flow column experiments were performed. Native microbial communities in subsurface sediments with no prior Cr(VI) exposure were shown to be capable of Cr(VI) reduction. In both the batch and column experiments, Cr(VI) reduction and loss from the aqueous phase were enhanced by adding high levels of both nitrate (NO3-) and organic C (molasses). Nutrient amendments resulted in up to 87% reduction of the initial 67 mg L(-1) Cr(VI) in an unsaturated batch experiment. Molasses and nitrate additions to 15 cm long unsaturated flow columns receiving 65 mg L(-1) Cr(VI) resulted in microbially mediated reduction and immobilization of 10% of the Cr during a 45-d experiment. All of the immobilized Cr was in the form of Cr(III), as shown by XANES analysis. This suggests that biostimulation of microbial Cr(VI) reduction in vadose zones by nutrient amendment is a promising strategy, and that immobilization of close to 100% of Cr contamination could be achieved in a thick vadose zone with longer flow paths and longer contact times than in this experiment.

Biodegradation, Environmental↗
PubMed2006

Importance of algal biomass to growth and development of Anopheles gambiae larvae.

We conducted experiments to investigate the importance of algal food resources for larval growth and adult emergence of Anopheles gambiae Giles s.s. in simulated larval habitats in Kenya, and in greenhouse and laboratory microcosms in the United States. In the first experiment, we used shading to reduce algal biomass, and because algal production and larval development might be a function of underlying soil nutrients, we crossed sun-shade treatments with soils of two distinct types collected near larval habitats. Shading reduced pupation rates and total adult biomass of An. gambiae by approximately 50%. Soil type had no significant effect on mosquito production, but it did significantly affect concentrations of phosphorus and chlorophyll a in the surface microlayer. In a subsequent experiment conducted in the greenhouse to reduce temperature differences found between the shaded and sunlit treatments, <1% of larvae in the shaded treatments reached the pupal stage. There was a marked reduction of chlorophyll a levels as a function of shading and larval density. In a third experiment, larvae receiving material harvested from sunlit surface microlayers performed as well as those receiving liver powder, whereas those receiving surface microlayer from shaded habitats suffered >90% mortality and failed to pupate. In a fourth experiment, glucose was added to shaded microcosms to stimulate bacterial activity in the absence of algae. Bacterial growth rates were 2 to 3 times higher, and larval development was enhanced in glucose-amended treatments. However, pupation rates and adult weights in glucose-amended shaded microcosms were still poor compared with those in nonamended sunlit microcosms. Overall, these results demonstrate the importance of algal biomass in the surface microlayers of larval habitats to development and adult production of An. gambiae.

Animal Feed↗