Die‐off of Escherichia coli in intact and disrupted cowpats

Diffuse pollution from grazed pastures is recognised as an important source of faecal microbial contamination of surface waters, and farmers urgently require mitigation options to reduce this impact of their farming activities. An obvious mitigation strategy is to reduce the size of the reservoir of faecal material in pastures during critical periods when overland flow is most likely. This work tested the hypothesis that smearing fresh cowpats to increase their surface area would increase exposure to solar radiation and increase desiccation, which in turn would accelerate the die‐off of Escherichia coli in the faecal material. The total solids content and E. coli concentration in intact and disrupted cowpats were monitored during the four seasons. The total solids content was significantly (P < 0.001) higher in the disrupted cowpats compared to the intact cowpats. This indicates that increased desiccation of the faecal material was successful. However, there was no significant difference (P > 0.05) in E. coli concentrations in the disrupted and intact cowpats. The physical disruption of cowpats did not therefore accelerate the die‐off of E. coli in the faecal material. The disrupted cowpats did appear to decompose faster than the intact cowpats. Mitigation options to reduce faecal micro‐organism losses in overland flow from grazed pastures will now need to focus on other methods to reduce the build up of reservoirs of faecal material on farm.     

Quantitatively tracing bacterial transport in saturated soil systems

Three isolates of Escherichia coli were labeled by their resistance to sodium azide and, separately, to novobiocin, nalidixic acid, and tetracycline. The strains exhibited a high degree of persistence in the soil environment and were recoverable on strain specific media at levels within the 95% confidence interval of the numbers covered on nonselective media. The E. coli strains were subsequently used to evaluate the events which would occur when a septic tank drainfield became submerged in a perched water table and effluentborne bacteria escaped into the groundwater. Field experiments were conducted by introducing the strains into horizontal lines installed into the A, B, and C horizons of a soil profile and transport was evaluated by collecting groundwater samples from 5 rows of piezometers (sampling six separate depth zones/row) located downslope from the injection lines. The major portion of subsurface transport of the bacterial populations occurred in specific zones in the soil profile and at an apparent maximum velocity of 17.0 cm min^?1. The maximum bacterial density in the groundwater, observed at each sampling distance downslope, was used to produce a mathematical relationship which described the overall decrease in numbers of organisms with increased distance through the soil. The potential health hazards which could occur by the subsurface transport of fecal organisms in relation to these experiments are discussed.     

Fate of gram-negative bacterial biomass in soil—mineralization and contribution to SOM

Soil microorganisms contribute to the formation of non-living soil organic matter (SOM) by metabolic transformation of plant-derived material. After cell death, their biomass components with a specific molecular character become incorporated into SOM imprinting its chemical properties, although this process has not yet been quantified. In order to elucidate the contribution to SOM formation, we investigated the fate of gram-negative bacterial model biomass (Escherichia coli usually introduced into soil with manure or feces) during incubation of soil with isotopically (¹³C) and genetically (lux gene) labeled cells. The decline of living cells was monitored by the loss of bioluminescence. The carbon turnover and mineralization was balanced by bulk soil stable isotope analysis, and the persistence of nucleic acids was investigated by PCR amplification of the lux gene. During incubation, the number of viable E. coli cells decreased rapidly (99.9% within the first 42 d) serving as substrate for other microorganisms or for the formation of SOM, and bioluminescent cells could only be detected during the first 56 d. However, the lux gene was still detected after 224 d, which indicates stabilization of DNA in SOM. Although the survival of E. coli in soil is limited, only about 65% of the added labeled biomass carbon was mineralized to ¹³CO₂ and 51% remained in soil after 224 d with an average ¹³C recovery of 117%. The amount of ¹³C found in the PLFA representative of living cells had decreased to 25% of the initial value, suggesting a proportional decrease of the ¹³C in the soil microbial biomass. The extent of this decrease is higher than the mineralization of the bulk E. coli C and thus the difference of around 25% has to be stabilized as metabolites, or in non-living SOM. The data provide evidence that the genetic information and a considerable part of the carbon from dying bacterial biomass were retained in both the soil microbial food web and in non-living SOM.     

Escherichia coli and Enterococci Attachment to Particles in Runoff from Highly and Sparsely Vegetated Grassland

Limited data on microbial partitioning between the freely suspended and particulate attached phases during transport along overland flow pathways have resulted in high uncertainty in bacterial fate and transport models and the application of these models to watershed management plans. The objectives of this study were to examine differences in attachment between E. coli and enterococci in runoff from plots with highly and sparsely vegetated grassland; investigate relations between flow regime, total suspended solids, and E. coli and enterococci attachment; and identify the particle size categories to which the attached cells were associated. Two rainfall simulations were conducted on large field plots 3 m wide by 18.3 m long with highly and both highly and sparsely vegetated covers and treated with standard cowpats. Results from the first experiment representing pasture with highly vegetated cover indicate that the majority of E. coli and enterococci are transported from the fresh manure source in the unattached state with only 4.8% of E. coli and 13% of enterococci associated with particles. The second experiment which compared partitioning in runoff from both highly and sparsely vegetated covers found lower bacterial attachment rates: the average E. coli percent attached was 0.06% from plots with highly vegetated cover and 2.8% from plots with sparsely vegetated cover while the corresponding values for enterococci were 0.98% and 1.23%, respectively. The findings from this study provide the first set of data on bacterial partitioning in overland flow from large field plots, and results may be helpful for parameterizing water quality models and designing conservation practices.     

S. typhimurium and E. coli O157:H7 retention and transport in agricultural soil during irrigation practices

During animal waste agricultural applications, the major concern is pathogen spreading, which may contaminate surface water and groundwater. Among the pathogenic microorganisms found in animal waste, Salmonella typhimurium and Escherichia coli O157:H7 are of particular concern. When transported in sub‐surface agricultural soil, S. typhimurium and E. coli O157:H7 are captured at the air–water–sediment interfaces through physical interactions. Because in situ colloids contribute to the formation of air–water–sediment, their mobilization affects the transport of S. typhimurium and E. coli O157:H7. The impact of irrigation rates on in situ colloid mobilization and S. typhimurium and E. coli O157:H7 transport was investigated in intact soil columns collected from an agricultural site in Gadsden County of Florida, USA. The columns were irrigated with sterilized nano‐pure deionized water to mobilize the colloids in the soil by stepwise increases in flow rate. For each flow rate, after colloids were mobilized and steady state was reached, S. typhimurium and E. coli O157:H7 were introduced. The cumulative amount of released in situ colloids increased linearly with the irrigation rates (R² = 0.986–0.996) and transport of the bacteria was enhanced after colloid mobilization. Interactions of the bacteria with the sediments and the air‐water interface were characterized: these played an important role in controlling S. typhimurium and E. coli O157:H7 retention in soil.     

The Transport of Escherichia coli Through Freeze-Fractured Clay Soil

Little is known about the transport of microorganisms through freeze-fractured clay soils. Normally consolidated clay (NCC) and compacted clay (CC) columns (representing a natural clay barrier and a compacted barrier, respectively) were exposed to six consecutive freeze–thaw cycles and permeated for 21 days with an Escherichia coli cell suspension (approximately 1?×?10⁷ colony forming units (CFU)/mL) containing a 2.1-mM bromide tracer. An unfractured sand column was also examined for comparison with the clay columns. While no E. coli was detected in the effluent of both untreated NCC and CC control clay columns, a relatively low density of E. coli (between 228 and 270 CFU/mL compared to 1?×?10⁷ CFU/mL in the influent) was first detected in the effluent of the freeze-fractured NCC and CC columns at 0.29 and 0.31 pore volumes (or at 5.4 and 4.1 h), respectively. It took 11 min for a full breakthrough of E. coli through the sand column, but only about 0.1% of the influent E. coli density was detected in the effluents of the freeze-fractured NCC and CC columns at day 21. These observations show that despite the high bacterial retention capacity of the freeze-fractured clay columns, the fractures were large enough for the E. coli to flow through. Based on batch sorption tests and the permeation data, it is estimated that 18%, 7%, and 84% of the freeze-fractured NCC, CC, and sand columns would be exposed to the influent, respectively, under a full E. coli breakthrough condition. Our data show that the high bacterial retention capacity of clay barriers can be compromised by freeze–thaw conditions.     

Preferential Attachment of Escherichia coli to Different Particle Size Fractions of an Agricultural Grassland Soil

This study reports on the attachment preference of a faecally derived bacterium, Escherichia coli, to soil particles of defined size fractions. In a batch sorption experiment using a clay loam soil it was found that 35% of introduced E. coli cells were associated with soil particulates >2 μm diameter. Of this 35%, most of the E. coli (14%) were found to be associated with the size fraction 15–4 μm. This was attributed to the larger number of particles within this size range and its consequently greater surface area available for attachment. When results were normalised with respect to estimates of the surface area available for bacterial cell attachment to each size fraction, it was found that E. coli preferentially attached to those soil particles within the size range 30–16 μm. For soil particles >2 μm, E. coli showed at least 3.9 times more preference to associate with the 30–16 μm than any other fraction. We report that E. coli can associate with different soil particle size fractions in varying proportions and that this is likely to impact on the hydrological transfer of cells through soil and have clear implications for our wider understanding of the attachment dynamics of faecally derived bacteria in soils of different compositions.     

Survival of faecal coliforms in four different types of sludge-amended soils in Botswana

This study aimed at identifying the factors that affected the survival of faecal coliforms as measured by E. coli in four types of soils in Botswana which were amended with sewage sludge. Physico-chemical and biochemical properties and coliform population in the different soils, sludge, and soil/sludge mixtures were determined after sampling, on composing the soil/sludge mixtures, and 90 days after composing. Coliform population in the different soil/sludge mixtures decreased by about 90% after 90 days. The age of the sludge used and the rate at which it was applied to the soils initially determined the population of E. coli in each soil/sludge mixture, but after 90 days, differences inherent in the different soil types were the main determinants of the E. coli population. Percentage reduction of coliforms in Type 1 sludge mixtures were lower (38%) than in Type 2 sludge mixtures (57%). Up to 79.8% of the reduction observed in E. coli population in the Barolong luvisol-sludge mixtures after 90 days was caused by reduction in pH and moisture content, while 72.6%, 84.5% and 55.1% of the reduction in E. coli population in Tuli luvisol-, arenosol- and vertisol-sludge mixtures, respectively, was accounted for by the reduction in moisture content and Olsen P concentration. Coliform survival rates varied with soil types being 12%, 6.4%, 5.3% and 5.8% for the vertisol, arenosol, Barolong luvisol and Tuli luvisol, respectively. A minimum period of 90 days should be allowed between when sludge is applied to similar soils and when seeds are sown. The exact period should, however, be determined by the properties of the soil with fine-textured soils requiring a longer period than coarse-textured soils.     

Simulating transport of E. coli derived from faeces of grazing livestock using the macro model

Abstract. Coliforms such as Escherichia coli and E. coli O157 are present in faeces deposited on the ground by grazing livestock, which gives rise to environmental concerns about the consequences of their transport in soil water draining to rivers, lakes, groundwater, water supplies and bathing waters. Following a similar study in relation to slurry spreading (Soil Use and Management 2003; 19, 321–330), a two‐stage approach was adopted to using the dual‐porosity contaminant transport model macro to simulate processes by which E. coli microorganisms from grazing livestock (sheep) pass through the soil to receiving waters via field drains. First, model parameter values were selected to reproduce experimental measurements showing rapid flows of the organisms by macropore flow without trapping in smaller pores. However, because of the large number of parameters and likely experimental errors, the set of values chosen, although plausible, is not necessarily unique and so any predictions should be considered provisional pending validation. Second, a series of predictive simulations was carried out to test the influence of soil and weather conditions on losses to field drains during grazing. These showed that E. coli losses were influenced almost entirely by the soil water content at the time of grazing, rising to a high level during grazing in wet conditions, but low or zero under dry conditions. In contrast, rainfall at the time of grazing had almost no consistent effect, other than large losses on the occasional days with over 20 mm of rain. Overall losses for a period of grazing were generally small during summer, but rose to a high level if grazing continued into autumn, due to the increase in soil water content. This demonstrates that there would probably be substantial reductions in the environmental risks of water pollution by E. coli and other faecal microorganisms if continuous grazing were stopped around early September and replaced by grazing on dry days only.     

Genetic variation in desiccation tolerance of Dendrobaena octaedra cocoons originating from different climatic regions

The aim of this study was to examine genetic variation in desiccation tolerance in cocoons of the parthenogenetically reproducing earthworm Dendrobaena octaedra by comparing populations originating from different geographic regions (Denmark, Norway and Finland), representing large differences in precipitation and temperature. In one experiment, the tolerance of the three populations to increasing desiccation stress in the range from 100 to 91.6% relative humidity (RH) was examined, aiming to represent ecologically relevant RH values. In a second experiment, the effect of cocoon size on desiccation tolerance was investigated at 92.3% RH in the same three populations. There were highly significant differences in desiccation tolerance between populations, indicating a high genetic differentiation of this trait in D. octaedra. Cocoons from Denmark were much more sensitive (71±14% mortality at 91.6% RH) than cocoons from Norway (21±4% mortality) and Finland (4±5% mortality). Cocoons of worms from Finland and Norway were significantly larger than cocoons produced by worms from Denmark suggesting that cocoons from Denmark lost water at a higher rate when subjected to low humidity. Assuming that slow dehydration is necessary for physiologically based protection mechanisms it may be expected that desiccation tolerance is positively correlated with cocoon size. However, within each of the populations cocoon fresh weight did not have any significant impact on desiccation tolerance. When all populations were pooled there was a significant positive effect of cocoon fresh weight on desiccation tolerance, explaining about 20% of the total variation (linear regression). It seems therefore that genetic variation of desiccation tolerance in D. octaedra cocoons is related to variation in both cocoon size and other, physiologically based tolerance mechanisms.     

Habitat moisture availability and the local distribution of the Antarctic Collembola Cryptopygus antarcticus and Friesea grisea

Population densities of the Collembola Cryptopygus antarcticus and Friesea grisea were compared in two maritime Antarctic habitats with different moisture availability. C. antarcticus was absent from the drier rock platform habitat, where F. grisea was the only collembolan collected. In contrast, the sand/pebble habitat on East Beach had greater moisture availability, and C. antarcticus dominated the arthropod community, with juveniles (individuals <1 mm length) representing 58% of the population. The hygropreference characteristics of F. grisea were determined in relative humidity (RH) gradients (12-98% RH) at 10 and 20 °C. F. grisea demonstrated a stronger preference for 98% RH conditions than C. antarcticus, suggesting that the former species is less likely to vacate moist refuges when available. The movement of both species was also monitored at 10 and 15 °C under conditions of 33, 75 and 100% RH. C. antarcticus was more active than F. grisea at both temperatures, and its movement increased at a greater rate as a consequence of reduced RH. The limited desiccation tolerance of C. antarcticus, combined with the increased water loss that would result from its continued movement under declining RH conditions, suggests this species is not well suited to drought-prone environments. In contrast, the reduced movement and ‘risk averse’ behavioural strategy of F. grisea, i.e. taking advantage of moist refuges when available, facilitates water conservation between precipitation/habitat rehydration events. This study provides the first evidence that moisture availability and habitat structure are potential habitat segregation mechanisms between these two Antarctic Collembola.     

Vadose Zone Microbial Transport Below At-Grade Distribution of Wastewater Effluent

The attenuation of Escherichia coli and total coliform from secondary treated wastewater effluent under two “at-grade” effluent distribution systems was evaluated in a sandy silt vadose zone in a cold climate. The two at-grade distribution lines had different designs and hydraulic loading rates. Effluent transport was examined using chloride as a tracer. Coliform fate was evaluated relative to the chloride using a combination of in situ pore water sampling and destructive soil sampling, combined with the observation of a dye tracer along excavation sidewalls. Although bacteria attenuation in the subsoil appeared to decrease during colder, winter temperatures (likely due to decreased viability and decreased predation), the subsoil provided about a four log reduction in E. coli over 90 cm of vertical transport. Horizontal transport of bacteria (up to 1.5 m from the line) was likely aided by flow on top of a microbial biomat observed at the soil surface. Both the subsurface dye patterns and the E. coli sampling suggested less preferential flow occurred below the lower loading rate design. At-grade distribution of secondary treated wastewater appears to be a viable alternative to conventional distribution fields at sites with similar climate and soils.     

Survival of Escherichia coli in soil with modified microbial community composition

Understanding the survival and persistence of Escherichia coli in soil with different microbial composition is essential for the accuracy of water quality assessment and microbial source tracking. This microcosm experiment was conducted to investigate the survival pattern of three E. coli strains (originated from soil, dog feces and human feces, separately) in soil with modified microbial community composition. Bile salt No. 3 (BS3) of progressively increased density (0.05%, 0.15%, 0.30% and 0.50%) was added into sandy loam soils and incubated for 90 days. Laboratory cultured E. coli were then inoculated into soil and incubated for another 150 days to monitor their survival pattern. Change of bacterial community diversity by BS3 was detected by both cultivation based and cultivation independent (PCR-Denaturing Gradient Gel Electrophoresis) methods. In general, progressively increased BS3 concentration resulted in decreased CFU counts both at 10 days and 90 days incubation. DGGE analysis indicated only a slight change in bacterial community composition at 10 days but a significant change at 90 days. Cluster analysis suggested that BS3 treatment grouped separately from controls. Survival of E. coli in soil was significantly influenced by the complexity of the microbial community, as die-off rate of E. coli progressively declined with the reduction of microbial community diversity. Differential survival of E. coli under different soil microbial stress highlights the importance of incorporating biotic factors in predictive models for water quality management and microbial source tracking study.     

Relative risk of surface water pollution by E. coli derived from faeces of grazing animals compared to slurry application

Abstract. This article examines some of the factors that influence the relative risk of Escherichia coli pollution of surface waters from grazing animals compared to cattle slurry application. Drainage water from pipe‐drained plots grazed with sheep (16 sheep + lambs per hectare) from 29 May to 17 July 2002 had average E. coli counts of 11 c.f.u. mL^?1 or 0.4% of estimated E. coli inputs over the grazing period. Drainage water from plots on the same site treated with cattle slurry (36 m³ha^?1 on 29 May 2002) had lower average E. coli counts of 5 c.f.u. mL^?1 or 0.03% of estimated faecal input. Sheep (16 lambs per hectare) grazing under cooler, moister conditions from 24 September to 3 December 2001 gave drainage water with much higher average E. coli counts of 282 c.f.u. mL^?1 or 8.2% of estimated input, which is more than twice the average E. coli counts previously reported under such conditions (Vinten et al. 2002 Soil Use and Management 18, 1–9). Laboratory studies of runoff from soil slabs after slurry application showed that the mobility of E. coli in surface soil decreased with time, suggesting that increased attachment to soil or migration to ‘immobile’ water also provides at least part of the physical explanation for the relatively higher risk of pollution from grazing animals compared with slurry. Sampling for E. coli in field drainflow and in streamwater during a storm event in the predominantly dairy Cessnock Water catchment, Ayrshire, Scotland supported the hypothesis that E. coli transport is linked to grazing animals. For a 7‐mm rainfall event, roughly 14% of the estimated daily input from grazing livestock was transported to the river, even though little slurry spreading had occurred in the catchment in the previous month. Spot sampling of field drains in grazed fields and silage fields in the same catchment also showed that grazing animals were the principal source of E. coli and faecal streptococci.     

Effects of Aeration,Molasses, Kelp,Compost Type,And Carrot Juice on the Growth of Escherichia Coli in Compost Teas

Growth of a nonpathogenic E. coli strain (K12- MG1655, ATCC 700926) in aerated and nonaerated compost teas containing molasses, kelp and carrot juice was examined. Teas were prepared using four different compost types that had undetectable levels of indigenous E. coli. Three of the composts were produced by turn pile windrow composting method using dairy, swine and horse manure as feedstock, while the fourth, a vermicompost, was produced by feeding separated dairy solids to worms Eisenia feotida. Molasses and kelp enhanced the growth of E. coli in inoculated teas and the E. coli density was positively correlated with nutrient concentrations ranging from 0.1 to 8.0 g/L. Irrespective of the presence of molasses and kelp, E. coli was not detected in noninoculated teas. Even though E. coli is a facultative anaerobe, its growth was significantly higher in nonaerated teas than in aerated teas. Without aeration, dissolved oxygen in teas declined rapidly and fell below 0.1 mg/L within 20 h, whereas continuous aeration at 0.8 L/min maintained an aerobic condition (> 5 mg/L dissolved oxygen) in teas during the 48 h brewing period. The pH values of nonaerated teas were significantly lower than those of aerated teas and were always slightly acidic. E. coli growth in different compost types was significantly different. The density of E. coli was lowest in teas made with vermicompost and highest in teas made with swine manure compost. E. coli proliferations in both aerated and nonaerated swine manure compost teas were inhibited by carrot juice. Carrot juice lowered dissolved oxygen in aerated teas. The total bacterial densities in noninoculated compost teas were not reduced by carrot juice.     

Discharge of Escherichia Coli from Agricultural Surface and Subsurface Drainage Water: Tillage Effects

Drainage water from agricultural fields with applied manure can degrade the bacterial quality of surface and groundwater. The impact of conventional tillage (CT) and zero tillage (ZT) practices on Escherichia coli (E.coli) discharge through artificially drained soils is not well understood. Consequently, two field trials were conducted during 2002–2004. The first trial involved fall applications of beef manure while the second involved spring applications of dairy manure. Both surface and subsurface drainage water were monitored in the first trial while only subsurface drainage water was monitored in the second. Under fall applied beef manure (trial 1), no differences (p?>?0.05) were observed in E.coli concentrations (cfu/100 ml) in combined drainage water under both tillage systems. However, during 2003–2004, subsurface drainage water under ZT had higher E.coli concentrations and loads than drainage water under CT. When the combined (surface + subsurface) annual E.coli loads were considered, CT loads were greater than ZT during 2002–2003 with an opposite situation during 2003–2004. Overall, annual E.coli loads were similar under ZT (4.7?×?10¹⁰ cfu/ha) and CT (4.8?×?10¹⁰ cfu/ha). Spring dairy manure application (trial 2) produced significant (p?>?0.03) tillage effect on E.coli loads in subsurface drainage water only during the second year. During the study period, ZT plots (1.55?×?10¹⁰ cfu/ha) discharged 5× more E.coli than CT (0.23?×?10¹⁰ cfu/ha). A longer duration of ZT practices resulted in higher subsurface flow volumes and subsequently greater loads of E.coli discharge in both trials.     

Protozoan predation of Escherichia coli in hydroponic media of leafy vegetables

Multiple outbreaks of food poisoning associated with fresh vegetable consumptions have occurred in many countries. Numerous reports have described human pathogenic bacteria, such as Escherichia coli O157:H7 and Salmonella spp., that can internalize into fresh vegetables via root or leaf surfaces. While attempting to obtain the threshold concentration of internalization of E. coli inoculated into hydroponic medium during vegetable cultivation, we observed a rapid decrease in E. coli numbers. In the present study, we determined that the rapid decline in E. coli was not due to a physiological change into a viable but non-culturable (VNC) state. The population crash was instead caused by true bacterial death, as the rapid descent was also confirmed by micro-colony fluorescence in situ hybridization, a culture-independent method that can detect VNC cells. We next monitored the number of E. coli inoculated into intact or filter-sterilized hydroponic medium after cultivation of various types of plants. We found that the number of E. coli in intact hydroponic medium decreased markedly, whereas the level in filter-sterilized hydroponic medium was completely unchanged. This result suggests that biotic factors were present that could be eliminated by filtering. Robust predation of E. coli by protozoa (ciliates and flagellates) was observed using fluorescently labeled bacteria incorporated into the hydroponic medium. Finally, morphological identification of flagellates by scanning electron microscopy revealed the presence of a species of Stramenopiles. These findings suggest the importance of protozoa as bacterial feeders in hydroponic systems and hence the use of these organisms as potential control agents of human pathogenic bacteria.     

Survival of <Emphasis Type="Italic">Escherichia coli</Emphasis> O157:H7 in various soil particles: importance of the attached bacterial phenotype

The risk of enteropathogens to food and water is highly dependent on their survival in soil environments. Here, the effects of soil type, particle size, the presence of natural organic matter (NOM) or Fe/Al (hydro)oxides on pathogenic Escherichia coli O157:H7 survival in sterilized soil particles were assessed through survival, attachment, metabolic activity, and qRT-PCR analyses. The abundance of inoculated E. coli O157:H7 in Brown soil (Alfisol) particles increased 0.6–1.4 log₁₀ CFU/g within 3 days (except for NOM-stripped clay), while that in Red soil (Ultisol) particles decreased rapidly in 8 days post-inoculation. Additionally, survival of bacteria was significantly enhanced when Fe/Al (hydro)oxides had been removed from Red soil particles. For the two soils, E. coli O157:H7 survived the longest in NOM-present clays and the bacterial adenosine 5′-triphosphate (ATP) levels were 0.7–2.0 times greater in clays than in sands and silts on day 8. Moreover, clays were more effective than silts and sands in binding cells and changing the expressions of acetate pathway-associated genes (pta and ackA). For silts and sands, E. coli O157:H7 decayed more rapidly in the presence of NOM and similar trends of bacterial ATP levels were observed between NOM-stripped and NOM-present soil particles, indicating that the primary role of NOM was not as a nutrient supply. These findings indicate that soil particles function mainly through attachment to change the metabolic pathway of E. coli O157:H7 and ultimately impact the survival of bacterial pathogens in soils.     

Influence of soil properties on the vertical movement of genetically-marked Pseudomonas fluorescens through large soil microcosms

Summary Vertical translocation of the introduced transposon Tn5-tagged Pseudomonas fluorescens cells was studies after irrigation of 50-cm long soil columns of loamy sand. The soil in the columns was slowly brought to saturation using groundwater, and enough water was then slowly added to permit collection of the percolated water. Introduced bacteria were transported to lower soil layers to a significantly higher degree in undisturbed soil cores than in repacked cores; water transport was hampered in both core types due to high soil bulk densities. Soil bulk density affected the degree of transport of the introduced cells; progressively more cells were translocated to deeper soil layers and into the percolation water at decreasing soil bulk densities. Repeated percolation of soil at a bulk density of 1.25 caused an increase in Tn5-tagged cell numbers in the lower soil layers and in the percolated water. Further, cells initially introduced into a dry (5.3% moisture) soil were translocated to a lesser extent than cells introduced into a wetter (13% moisture) soil. Finally, wheat roots enhanced the water-induced transport of introduced cells to the 40- and 50-cm deep soil layers and into the effluent, but not to the remaining soil layers. Large soil columns such as those used in the present study are useful in assessing the transport and survival of introduced bacterial cells in soils under a variety of simulated environmental conditions.     

Chemical and Microbiological <Emphasis Type="Italic">in situ</Emphasis> Characterization of Benthic Communities in Sediments with Different Contamination Levels

Background  River sediments are natural habitats of complex bacterial and fungal communities and therefore play a decisive role in the mineralization process of organic matter in freshwater systems. By means of comparative temporal and spatial analyses of microbial communities, the in situ impact of anthropogenically generated pollutants on these biofilm associations can be assessed and discriminated from seasonal variations. Aim  The aim was the adaptation of hybridization with fluorescently labelled rRNA-targeted oligonucleotides (FISH) for the in situ characterization of the structural and functional diversity of native microbial communities in complex lotic sediments. The impact of qualitatively and quantitatively different water pollutants on the microbial diversity, metabolic potential, and relative abundance of characteristic bacterial groups was assessed by oligonucleotide probes on different phylogenetic levels. In particular, sulfate reducing bacteria (SRB) were investigated to evaluate their potential applicability as microbial biomonitors in sediments. Methods  Sediment samples from the German lowland rivers Elbe and Oder were investigated over 12 months with regard to physico-chemical parameters and the composition of the attached microbial communities. Mechanical treatment including ultrasonification and sagitation under aerobic conditions combined with the use of pyrophosphate ensured the equal dispersion of fixed microbial cells within the sediment samples. The optimized whole-sediment FISH-technique was combined with an improved cell extraction procedure and applied, due to the specific grain size fraction distribution, at the different sampling sites. Resultsand discussion  Up to 85.6% of the total bacterial cell counts as determined by DAPI (4’, 6-diamidino-2-phenylindole) staining could be successfully monitored by the eubacterial oligonucleotide probe set EUB338, EUB338-II and EUB338-III, simultaneously indicating a high proportion of Eubacteria and the high metabolic potential of the bacterial community. Desulfobacteriaceae could be detected by the specific probe SRB385Db in various relative percentages ranging from 2.4 to 16.0% of the total bacterial cell counts. The total number of bacteria and the metabolic potential of sediment related bacteria were barely affected by the different pollution pattern of the sampling sites. Conclusions  The pre-treatment step as conducted by cell extraction as well as the FISH hybridization procedure was successfully optimized to the specific conditions present within freshwater sediments. Beside seasonal variations, particularly occurring at hydrologically influenced sites, sampling sites with different pol lution levels could be successfully distinguished by the relative abundance of Desulfobacteriaceae used as microbial indicator organisms. Outlook  The integration of ongoing insights into pollution induced changes of natural bacterial consortia should result in a system of ecotoxicological classes representing the different ecological status of riverine systems. Physiological directed methods like Community Level Physiological Profiling (CLPP) or Pollution Induced Community Tolerance (PICT), and structural techniques as FISH or microarrays should be used to investigate the influence of harmful substances on the biodiversity in natural microbial sediment communities.