Marianne Bischoff

Surface and subsurface microbial biomass, community structure and metabolic activity as a function of soil depth and season

Microbial biomass, size and community structure along with an estimate of microbial activity and soil chemical parameters were determined at three depths in two soils (e.g. sandy loam Ultic Hapludalf and silt loam Mollic Hapludalf ) replicated three times under one winter and summer season. Microbial biomass and community structure were estimated from phospholipid-PO4 content and fatty acid methyl ester (FAME) measurements. Microbial activity and assimilative capacity were estimated using a 3 H-acetate incorporation into phospholipids and by incubating the soil samples at the average winter and summer temperatures, 3 and 20 ◦ C, respectively. We found that the size of the microbial biomass in both the surface and the subsurface soils was not significantly affected by the seasonal variation but activity increased by as much as 83% at the summer temperatures in the surface soil. We demonstrated using FAME analysis that for both soils seasonal changes in the subsurface microbial community occurred. These findings suggest that winter conditions will shift the population activity level in both the surface and subsurface systems and the biochemical structure of the community in the subsurface. In all cases, the inorganic chemical properties of the soil, as a function of season, remained constant. The greatly increased activity of microbial population at the higher temperature will favor the capacity of the system to utilize nutrients or organic materials that may enter soil. During low temperature seasons the capacity of either surface or subsurface soils to assimilate materials is generally diminished but the reduction reflects changes in metabolism and not a reduced biomass size.

Association of Microbial Community Composition and Activity with Lead, Chromium, and Hydrocarbon Contamination

ABSTRACT Microbial community composition and activity were characterized in soil contaminated with lead (Pb), chromium (Cr), and hydrocarbons. Contaminant levels were very heterogeneous and ranged from 50 to 16,700 mg of total petroleum hydrocarbons (TPH) kg of soil−1, 3 to 3,300 mg of total Cr kg of soil−1, and 1 to 17,100 mg of Pb kg of soil−1. Microbial community compositions were estimated from the patterns of phospholipid fatty acids (PLFA); these were considerably different among the 14 soil samples. Statistical analyses suggested that the variation in PLFA was more correlated with soil hydrocarbons than with the levels of Cr and Pb. The metal sensitivity of the microbial community was determined by extracting bacteria from soil and measuring [3H]leucine incorporation as a function of metal concentration. Six soil samples collected in the spring of 1999 had IC50 values (the heavy metal concentrations giving 50% reduction of microbial activity) of approximately 2.5 mM for CrO42− and 0.01 mM for Pb2+. Much higher levels of Pb were required to inhibit [14C]glucose mineralization directly in soils. In microcosm experiments with these samples, microbial biomass and the ratio of microbial biomass to soil organic C were not correlated with the concentrations of hydrocarbons and heavy metals. However, microbial C respiration in samples with a higher level of hydrocarbons differed from the other soils no matter whether complex organic C (alfalfa) was added or not. The ratios of microbial C respiration to microbial biomass differed significantly among the soil samples (P < 0.05) and were relatively high in soils contaminated with hydrocarbons or heavy metals. Our results suggest that the soil microbial community was predominantly affected by hydrocarbons.

Microbial community analysis of soils contaminated with lead, chromium and petroleum hydrocarbons.

The impact on the microbial community of long-term environmental exposure to metal and organic contamination was investigated. Twenty-four soil samples were collected along a transect dug in soils contaminated with road paint and paint solvents, mainly toluene. Chemical analysis along the transect revealed a range from high to low concentrations of metals (lead and chromium) and organic solvent compounds. Principal components analysis of microbial community structure based on denaturing gradient gel electrophoresis of the V3 region of the 16S rRNA gene and fatty acid methyl esters derived from phospholipids (phospholipid fatty acid analysis) showing samples with similar fingerprints also had similar contaminant concentrations. There was also a weak positive correlation between microbial biomass and the organic carbon concentration. Results indicated that microbial populations are present despite some extreme contaminant levels in this mixed-waste contaminated site. Nucleotide sequence determination of the 16S rRNA gene indicated the presence of phylogenetically diverse bacteria belonging to the α-, β-, γ-, and δ-Proteobacteria, the high and low G + C Gram-positive bacteria, green nonsulfur, OP8, and others that did not group within a described division. This indicates that soils contaminated with both heavy metals and hydrocarbons for several decades have undergone changes in community composition, but still contain a phylogenetically diverse group of bacteria (including novel phylotypes) that warrant further investigation.

Long-term effects of chromium and lead upon the activity of soil microbial communities

The inhibitory effects of heavy metals upon microbial populations and the factors limiting microbial activity and growth were determined in soils that had been contaminated with lead (Pb) or chromium (Cr) for more than 40 years. Total Cr and Pb concentrations were 260,000 and 10,000 mg kg −1 soil in Cr- and Pb-contaminated soils, respectively. The tolerance of bacteria extracted from soil particles to Pb or Cr was assayed by measuring the incorporation of 3 H -leucine into macromolecules at a series of CrO4 2− or Pb 2+ concentrations. IC50 (heavy-metal concentration giving 50% reduction of microbial 3 H -leucine incorporation compared to the control) for the two metals was similar in the two soils, despite their differences in metal contamination; the IC50 values were 4 and 0.02 mM for CrO4 2− and Pb 2+ , respectively. Stimulation of microbial activity and biomass by organic C was measured in microcosms over 56 days. The adverse effects of Cr and Pb were characterized in terms of the ratio of microbial biomass C to soil organic C, basal respiration per unit microbial biomass (qCO2), and the ratio of substrate-responsive respiration to microbial biomass C (substrate-responsive qCO2). Ratios of microbial biomass C to soil organic C were small: 0.42% in Cr-contaminated soil and 0.36% in Pb-contaminated soil. Values of qCO2 and substrate-responsive qCO2 were significantly higher in Pb- than in Cr-contaminated soils ( P< 0.01). The microcosm results indicate that Cr and Pb decreased microbial activities and led to the accumulation of soil organic C, and that Pb posed greater stress to soil microbes than Cr.

Response of Soil Microorganisms to As-Produced and Functionalized Single-Wall Carbon Nanotubes (SWNTs)

The use of single-wall carbon nanotubes (SWNTs) in manufacturing and biomedical applications is increasing at a rapid rate; however data on the effects of a potential environmental release of the materials remain sparse. In this study, soils with either low or high organic matter contents as well as pure cultures of E. coli are challenged with either raw as-produced SWNTs (AP-SWNTs) or SWNTs functionalized with either polyethyleneglycol (PEG-SWNTs) or m-polyaminobenzene sulfonic acid (PABS-SWNTs). To mimic chronic exposure, the soil systems were challenged weekly for six weeks; microbial activities and community structures for both the prokaryote and eukaryote community were evaluated. Results show that repeated applications of AP-SWNTs can affect microbial community structures and induce minor changes in soil metabolic activity in the low organic matter systems. Toxicity of the three types of SWNTs was also assessed in liquid cultures using a bioluminescent E. coli-O157:H7 strain. Although decreases in light were detected in all treated samples, low light recovery following glucose addition in AP-SWNTs treatment and light absorption property of SWNTs particles suggest that AP-SWNTs suppressed metabolic activity of the E. coli, whereas the two functionalized SWNTs are less toxic. The metals released from the raw forms of SWNTs would not play a role in the effects seen in soil or the pure culture. We suggest that sorption to soil organic matter plays a controlling role in the soil microbiological responses to these nanomaterials.

The role of tree leaf mulch and nitrogen fertilizer on turfgrass soil quality

Abstract The influence of tree leaf amendment and N fertilization on soil quality in turfgrass environments was evaluated. Our objective was to assess changes in soil quality after additions of leaf materials and N fertilization by monitoring soil chemical and physical parameters, microbial biomass and soil enzymes. Established perennial ryegrass (Lolium perenne) plots were amended annually with maple (Acer spp.) leaves at three different rates (0, 2240, and 4480 kg ha–1 year–1) and treated with three nitrogen rates (0, 63, and 126 kg N ha–1 year–1). Tree leaf mulching did not significantly affect water infiltration or bulk density. However, trends in the data suggest increased infiltration with increasing leaf application rate. Tree leaf mulching increased total soil C and N at 0–1.3 cm depth but not at 1.3–9.0 cm. Extracted microbial phospholipid, an indicator of microbial biomass size, ranged from 28 to 68 nmol phospholipid g–1 soil at the 1.3–9.0 cm depth. The activity of β-glucosidase estimated on samples from 0–1.3 cm and 1.3–9.0 cm depths, and dehydrogenase activity estimated on samples from 1.3–9.0 cm were significantly increased by leaf mulching and N fertilizer application. Changes in microbial community composition, as indicated by phospholipid fatty acid methyl ester analysis, appear to be due to seasonal variations and did not reflect changes due to N or leaf amendment treatments. There were no negative effects of tree leaf mulching into turfgrass and early data suggest this practice will improve soil chemical, physical, and biological structure.

Environmental implications of nanomaterials: are we studying the right thing?

A fundamental lack of data on the potential impacts of carbon based nanomaterials on natural ecosystems currently exists. The gap between what we know about environmental impacts and new products that may contain nanomaterials continues to get wider especially related to knowledge about nanocomposites. In this paper we present ideas and concerns about the current state of knowledge on nanomaterials in the environment and present a number of points about what recent work has provided us about the novel materials.

Understanding the role of agricultural practices in the potential colonization and contamination by Escherichia coli in the rhizospheres of fresh produce.

To better protect consumers from exposure to produce contaminated with Escherichia coli, the potential transfer of E. coli from manure or irrigation water to plants must be better understood. We used E. coli strains expressing bioluminescence (E. coli O157:H7 lux) or multiantibiotic resistance (E. coli²(+)) in this study. These marked strains enabled us to visualize in situ rhizosphere colonization and metabolic activity and to track the occurrence and survival of E. coli in soil, rhizosphere, and phyllosphere. When radish and lettuce seeds were treated with E. coli O157:H7 lux and grown in an agar-based growth system, rapid bacterial colonization of the germinating seedlings and high levels of microbial activity were seen. Introduction of E. coli²(+) to soil via manure or via manure in irrigation water showed that E. coli could establish itself in the lettuce rhizosphere. Regardless of introduction method, 15 days subsequent to its establishment in the rhizosphere, E. coli²(+) was detected on the phyllosphere of lettuce at an average number of 2.5 log CFU/g. When E. coli²(+) was introduced 17 and 32 days postseeding to untreated soil (rather than the plant surface) via irrigation, it was detected at low levels (1.4 log CFU/g) on the lettuce phyllosphere 10 days later. While E. coli²(+) persisted in the bulk and rhizosphere soil throughout the study period (day 41), it was not detected on the external portions of the phyllosphere after 27 days. Overall, we find that E. coli is mobile in the plant system and responds to the rhizosphere like other bacteria.

Influence of fullerene (C60) on soil bacterial communities: aqueous aggregate size and solvent co-introduction effects.

Fullerene C60 nanoparticles are being used in broad range of applications. It is important to assess their potential impacts in the environment. We evaluated the effects of C60 introduced as aqueous suspensions of nC60 aggregates of different particle size or via organic solvents on soils with different organic matter contents in this study. Impacts of the application were evaluated by measuring total microbial biomass, metabolic activity and bacterial community structure. Results show that nC60 aggregates, introduced as an aqueous suspension, had size-dependent effects on soil bacterial community composition in the low organic matter system, but induced minimal change in the microbial biomass and metabolic activity in soils with both high and low organic matter contents. Fullerene C60, co-introduced via an organic solvent, did not influence the response of soil microbes to the organic solvents. Our results suggest that nC60 aggregates of smaller size may have negative impact on soil biota and soil organic matter may play a key role in modulating the environmental effect of nanomaterials.

Rapid and complete degradation of the herbicide picloram by Lipomyces kononenkoae.

An enrichment culture approach was used to isolate a pure culture of the yeast Lipomyces kononenkoae, which had the ability to grow on the herbicide picloram. The yeast rapidly and completely degraded 50 microg mL(-1) picloram by 48 h of growth. While L. kononenkoae was found to use both N atoms of picloram as a sole nitrogen source for growth, it failed to mineralize the herbicide or use it as a sole C source. Product analysis done using LC-ESI-MS indicated that biodegradation of picloram by L. kononenkoae proceeds via a didechlorinated, dihydroxylated, pyridinecarboxylic acid derivative. Our results are consistent with the hypothesis that the majority of picloram degradation in the soil is likely due to microbial catabolic processes.