Fate of Escherichia coli and Escherichia coli O157 in soils and drainage water following cattle slurry application at 3 sites in southern Scotland

Abstract. Slurry from farm animals may contaminate water supplies, rivers and bathing waters with faecal coliforms, such as Escherichia coli . Where animals harbour the O157 strain the hazard to human health is particularly high, but both the hazard level, and the low incidence and sporadic nature of the excretion of E. coli O157 make it difficult to study this strain under field conditions. The survival of total E. coli and of E. coli O157 were compared in the laboratory for two soils under controlled temperature and moisture. E. coli O157 die-off rate was the same as or quicker than for total E. coli . This result meant that field experiments studying the fate of total E. coli should give a satisfactory evaluation of the risk of water contamination by the O157 strain. In four field experiments at three sites, slurry containing total E. coli numbers of 2.2 × 10⁴ to 5.7 × 10⁵ colony forming units per mL (c.f.u. mL^–1) was applied to drained field plots. Field die-off was faster than expected from laboratory experiments, especially in one experiment where two weeks dry weather followed application. In all but this experiment, the first drain flow events after slurry application led to very high E. coli concentrations in the drains (10³ to 10⁴ c.f.u. mL^–1). E. coli O157 was present in the slurry used for two of the experiments (33 c.f.u. per 100 mL in each case). However the proportion of E.coli O157 was very low (about 1 in 10⁵) and it was not detected in the drainage water. After the first week E. coli drainage water numbers decreased rapidly but they were 1–10 c.f.u. mL^–1 for much of the sampling period after slurry application (1–3 months).     

Effect of Inorganic N Composition of Fertilizers on Nitrous Oxide Emission Associated with Nitrification and Denitrification

We studied the effect of repeated application (once every 2 d) of a fertilizer solution with different ratios of NH₄⁺ - and NO₃⁻-N on N₂O emission from soil. After the excess fertilizer solution was drained from soil, the water content of soil was adjusted to 50% of the maximum water-holding capacity by suction at 6 × 10³ Pa. Repeated application of NH₄⁺- rich fertilizer solution stimulated nitrification in soil more than NO₃⁻-rich fertilizer. Although the evolution of N₂O through nitrifier denitrification tended to increase with the repeated addition of a fertilizer solution rich in NH₄⁺ rather than in NO₃⁻, the contribution of nitrifier denitrification remained at levels of 20 to 36% of the total emission regardless of the inorganic N composition. The total emission of N₂O also tended to increase with the application of NH₄⁺- rather than NO₃⁻-rich fertilizer. It was suggested that the coupled process of nitrification and denitrification at micro-aerobic sites became important when fertilizer rich in NH₄⁺ was applied to soil under relatively aerobic conditions.     

Changes in the microbial community and physico-chemical characteristics of topsoils stockpiled during opencast mining

Abstract. Experiments on the effects of stockpiling soil on an opencast coal mine in Derbyshire showed that there were significant changes in the microbial community. Numbers of aerobic bacteria in stored soils ranged from 0.5 to 12.8 ± 10⁷ colony-forming-units (CFU)g^-1 with the smallest values being in the deepest parts of the oldest stores, whereas an adjacent undisturbed soil contained 6.6 ± 10⁷ CFU g^-1. There was a greater effect on the numbers of fungal spores, which ranged from 0.1 to 6.7 ± 10⁵ CFU g^-1 soil, all less than the 10 ± 10⁵ CFU g^-1 recorded for the undisturbed control soil. The number of fungal spores in the deepest part of the older soil stores was only 1/100 of the number in the undisturbed soil. This was mirrored by the biomass values, as determined by adenosine triphosphate (ATP) assay. Values of ATP ranged from 0.38 to 13.13 nmol g^-1 as compared to 5.8 nmol g^-1 in the undisturbed soil. All three of these microbiological properties decreased in value with both age and depth of storage. Neither anaerobic nor spore-forming bacterial numbers were greatly affected by storage.
The pH values tended toward neutrality in the deeper parts of the soil stores, and there was less organic matter in the older stores.     

Influence of the nature of clay minerals on the fixation of radiocaesium traces in an acid brown earth–podzol weathering sequence

The magnitude of radiocaesium fixation by micaceous clay minerals is affected by their transformation, which depends on weathering in soil. The net retention of radiocaesium traces was quantified by sorption–desorption experiments in the various horizons of four sandy soils forming an acid brown earth–podzol weathering sequence derived from sandy sediments and characterized by marked changes in mineral composition. The features of the 2:1 minerals of the four soils, resulting from an aluminization process in depth and a desaluminization process towards the surface, had a strong influence on Cs⁺ fixation. Beneath the desaluminization front, which deepens from the acid brown earth to the podzol, hydroxy interlayered vermiculite was dominant and the ¹³⁷Cs⁺ fixation was the weakest. At the desaluminization front depth, vermiculite was responsible for the strongest ¹³⁷Cs⁺ fixation. In the upper layers, smectite appeared in the podzolized soils and the ¹³⁷Cs⁺ fixation decreased. The magnitude in Cs⁺ fixation therefore appeared as a tracer of the transformation process affecting the 2:1 clay minerals in the acid brown earth–podzol weathering sequence. This magnitude was positively correlated with the vermiculite content of the studied soil materials estimated by the rubidium saturation method.     

Effects of organic matter and calcium on soil structural stability

The cationic bridging effect of the calcium ion (Ca²⁺) and the flocculating ability of clay and organic matter are crucial in the formation and stability of soil aggregates. They are therefore likely to influence the soil's saturated hydraulic conductivity ( K _s). We tested the individual effects of these factors on aggregate stability and related hydraulic properties, and studied the influence of clay mineralogy also. Samples from the surface (0–10 cm) of three contrasting soils in Trinidad were used. The soils were treated with three levels of Ca²⁺ and three levels of organic matter in a 3 × 3 × 3 factorial design and incubated for 14 days. Both aggregate stability and saturated hydraulic conductivity were influenced by all factor combinations. Interactions between soil type and Ca²⁺ revealed the importance of polyvalent cations in aggregate stability of soils with low activity minerals. The influence of organic matter varied with quantity; the more there was, the more stable the soil became, particularly in the soil containing little clay. Clay dispersion and slaking of expanding minerals occurred even with large additions of Ca²⁺ and organic matter, emphasizing the overall influence of mineralogy in determining the response of soils to stability treatments.     

Pollution of carbonate soils in a Mediterranean climate due to a tailings spill

The retention walls in a pond containing the residues from the pyrite mine of Aznalcóllar (southern Spain) broke open on 25 April 1998, spilling approximately 6 × 10⁶ m³ of polluted water and toxic tailings, which affected some 55 km². Drying and aeration of the tailings resulted in oxidation, forming an acidic solution with high pollutant contents, the effects of which were studied in a calcareous soil. The infiltration of this solution markedly affected only the first 12 mm of the soil, where strong acidification caused the weathering of the carbonates, and where the fine mineral particles were hydrolysed. The SO₄²⁻ ions in the acidic solution precipitated almost entirely at this depth, forming gypsum, hydroxysulphates and complex sulphates. The Fe³⁺ ions also precipitated there, mainly in amorphous or poorly crystallized forms, adsorbing to As, Sb, Tl and Pb dissolved in the acidic solution. The Al³⁺ ions, though partly precipitating in the acidic layer, accumulated mostly where the soil pH exceeded 5.5 (12–14 mm in depth). They did so primarily as amorphous or poorly crystallized forms, adsorbing to Cu dissolved in the acidic solution. The Zn²⁺ and Cd²⁺ ions accumulated mainly at pH > 7.0 (19–21 mm in depth), being adsorbed chiefly by clay mineral. After 15 months, only the first 20 mm of the soil were acidified by the oxidation of the tailings and most of the pollutants did not penetrate deeper than 100 mm. Consequently, the speed of the cleanup of the toxic spill is not as important as a thorough removal of tailings together with the upper 10 cm of the soil.     

Scaling the saturated hydraulic conductivity of an alfisol

Alfisols exhibit a high degree of spatial variability in their physical properties. As a result, it is difficult to use information on physical parameters measured at one location to model larger-scale hydrologic processes. In this study, the saturated hydraulic conductivity, K_S , of an Alfisol was determined on 109 undisturbed monoliths using the falling-head permeameter method. The model developed by Arya & Paris (1981) was used to calculate the pore volume from sand and clay fractions. Scaling factors were calculated from the measured K_s , sand pore-volume, clay pore-volume, clay content and effective porosity, using the similar media concept. Prediction of K_s of gravelly Alfisol using clay pore-volume is confounded by high gravel content which, when discounted, improves the prediction remarkably. The scaled mean saturated hydraulic conductivity K* for all horizons of the Alfisol was approximately l.0x 10⁻⁵ms⁻¹.     

Retarded leaching of nitrate measured in monolith lysimeters in south-east Nigeria

At Onne in South-east Nigeria, drainage water was collected from four monolith lysimeters and analysed for nitrate. The lysimeters contained an acid sandy loam. At the start of the first rainy season two lysimeters received urea labelled with ¹⁵NO₃^– and two received no nitrogen fertilizer; all four were uncropped in the first year.
The peak concentrations of ¹⁵NO₃^– and of unlabelled (soil) NO₃^– were found after 2.5 pore volumes of water had passed through the lysimeters. Using the same soil in the laboratory after fine sieving, the peak concentration of tritiated water was found at 1 pore volume whereas nitrate leaching was retarded. The pattern of nitrate leaching was well described by miscible and immiscible models which included an adsorption coefficient for nitrate. Over the 2 years 81.4% of the ¹⁵N added at the start of the first rainy season was recovered in the drainage water.     

Real-time imaging of nitrogen fixation in an intact soybean plant with nodules using 13N-labeled nitrogen gas

Real-time images of nitrogen fixation in an intact nodule of hydroponically cultured soybean ( Glycine max [L] Merr.) were obtained. In the present study, we developed a rapid method to produce and purify ¹³N-labeled radioactive nitrogen gas (half life: 9.97 min). ¹³N was produced from a ¹⁶O (p, α) ¹³N nuclear reaction. The target chamber was filled with CO₂ and irradiated for 10 min with protons at an energy of 18.3 MeV and an electric current of 5 μA, which was delivered from a cyclotron. All CO₂ in the collected gas was absorbed and removed with powdered soda-lime in a syringe and replaced with helium gas. The resulting gas was injected into gas chromatography and separated and a 35 mL fraction, including the peak of [¹³N]-nitrogen gas, was collected by monitoring the chromatogram. The obtained gas was mixed with 10 mL of O₂ and 5 mL of N₂ and used in the tracer experiment. The tracer gas was fed into the underground part of intact nodulated soybean plants and serial images of the distribution of ¹³N were obtained non-invasively using a positron-emitting tracer imaging system (PETIS). The rates of nitrogen fixation of the six test plants were estimated to be 0.17 ± 0.10 μmol N₂ h⁻¹ from the PETIS image data. The decreasing rates of assimilated nitrogen were also estimated to be 0.012 ± 0.011 μmol N₂ h⁻¹. In conclusion, we successfully observed nitrogen fixation in soybean plants with nodules non-invasively and quantitatively using [¹³N]N₂ and PETIS.     

Risk assessment of nitrate pollution in groundwater based on the environmental nitrogen-assimilation capacity of agricultural lands in Hokkaido

(pp. 17–24)
A trial calculation was performed of the environmental nitrogen-assimilation capacity and the amount of nitrogen input based on various statistical data, which were compiled from each city, town and village in Hokkaido prefecture. The relationship between the excess quantity of nitrogen, after nitrogen input, and the environmental nitrogen-assimilation capacity and the nitrate-nitrogen concentration of the groundwater was considered.
Environmental nitrogen-assimilation capacity = nitrogen output by the crops + acceptable level of residual nitrate in the soil profile.*
^*It is calculated by the amount of nitrate precipitation evapotranspiration ×10 mg L⁻¹.

• 1) 

  The average value of the environmental nitrogen-assimilation capacity in Hokkaido Prefecture was observed to be 183 kg ha⁻¹. The maximum and minimum values of the environmental nitrogen-assimilation capacity were 308 kg ha⁻¹ and 94 kg ha⁻¹, respectively. When the average value of the environmental nitrogen-assimilation capacity with respect to main agricultural land use was compared across municipalities, it was largely in the following order · grassland (218 kg ha⁻¹), upland (169 kg ha⁻¹), and paddy land (157 kg ha⁻¹).

       
  
  

Analysis of Potassium Uptake by Rice Roots Treated with Aluminum Using a Positron Emitting Nuclide, 38K

We investigate the effect of Al on K⁺ uptake by rice roots. Potassium-38 (³⁸K), a positron emitting nuclide (the half-life: 7.61 min), was used to trace K⁺ behavior. When a rice root was treated with 10μM Al for 24 h, the uptake of ³⁸K in the root was increased in the range of 1 to 2 cm from the root tip compared with that of the control sample. Because the root continued to grow without showing any damage of plasma membrane during the Al treatment, it was suggested that the ³⁸K uptake was not occurred through diffusion into the cells. The uptake of ³⁸K in both treatments, with/without Al, was decreased by VO₄^3- (inhibitor of H⁺-ATPase on plasma membrane) and DNP (H⁺ ionophore) treatment, which suggested that the K⁺ uptake was performed through an active transport, such as H⁺:K⁺ transport or H⁺ gradient promoted by an Al treatment.     

Selenite fixation by soil particle-size separates

The fixation of selenite by clay- (< 2 μm), silt- (2–20 μm) and sand-size (20–2000 μm) separates from two arable soils was examined in solutions of ⁷⁵Se-labelled sodium selenite using a Se/sample ratio of 1/10⁶. Size separates were isolated by ultrasonic dispersion and gravity sedimentation. Selenite fixation was determined after equilibration periods ranging from 5 min to 26 h. Hydrogen peroxide-treated samples were included to examine the effect of organic matter on selenite fixation capacity.
The relative distribution of native Se, C, dithionite/citrate-extractable Fe and Al between size separates was similar. Concentrations in clay were four to nine times higher than in whole soils, silt showing two to five times higher concentrations and sand being very low in Se, C, Fe and Al.
After 1 h, clay, silt and sand fixed 64–65%, 45–61% and <5% of the selenite added, respectively. The fixation on whole soils was 14–18%. After 1 day, fixation on clay, silt, sand and whole soil increased to 78–87%, 67–79%, 3–14% and 31–39%, respectively.
Hydrogen peroxide-treatment reduced the selenite fixation capacity of whole soil, silt and sand to very low levels. Fixation on peroxide-treated clay was in accord with values for pure clay minerals reported in the literature. Generally, the fixation capacity of peroxide-treated natural clay and pure clay minerals was only half that observed for intact clay-size separates, demonstrating the importance of organic matter in soil selenite fixation capacity.     

Short-term kinetics of residual wheat straw C and N under field conditions: characterization by 13C15N tracing and soil particle size fractionation

Summary A clear understanding of the short-term decomposition and fate of crop residues is necessary to predict the availability of mineral N in soil. The fate of ¹³ C¹⁵N-labelled wheat straw in a silty soil (Typic Hapludalf) was studied using particle size fractionation and in situ incubation in which the equivalent of 8 t dry matter per ha of straw was incorporated into the soil over 574 days. Soil samples were separated into five particle-size fractions by wet sieving after disruption of aggregates. The weight, C and N contents, and ¹³C and ¹⁵N atom excess of each fraction were determined. Straw-derived C disappeared rapidly from the > 2000-μm fraction with an estimated half-life of 53 'normalized' days (equivalent of 10°C and −0−01 MPA water potential). Straw-derived C appeared to be only temporarily stored in the intermediate fractions (1000–2000 and 200–1000 pm). The maximum net ¹³C accumulation in the 50–200-μm fraction was 4·4% of added ¹³C. Straw-derived C accumulated most rapidly and preferentially in the 50-μm fraction, which stabilized after 265 days and accounted for 70% of the residual ¹³C on day 574. Although there was more residual ¹⁵N than ¹³C, the distributions and kinetics of the two isotopes in the fractions were similar.     

Release of potassium from some benchmark soils of India

Release of potassium from 15 surface samples of benchmark Alluvial, Red and Black soils of India to 0.01 M solutions of BaCl₂, CaCl₂, NH₄Cl and NaCl was studied in soils either untreated or pretreated with 5 × 10⁻³ M KCl. In the untreated soils, the efficacy of the extractants declined in the sequence: BaCl₂ > NH₄Cl > CaCl₂ > NaCl. Cumulative K-release was greatest from Black soils, followed by Red and Alluvial soils. From soils pretreated with 5 * 10⁻³ M KCl, more K was released than retained, and more 'native' K was released than that from untreated soils. Increase in the release of 'native' K decreased in the sequence: Red > Alluvial > Black soils. The amounts of surface and internal K, desorption rate constants and parabolic diffusion constants were calculated from K release to the various electrolytes.     

Metal-humus complexes in A horizons of Thai and Korean red and yellow soils

Carbon, Al and Fe (C_pyr, Al_pyr and Fe_pyr) were extracted with 0.1 m Na₄P₂O₇ from 26 A horizon samples of tropical Thai and temperate Korean soils (Ultisols, Alfisols, Oxisols and Inceptisols). The soils, except for one Thai Inceptisol, had similar total C (0.35–3.29%) and C_pyr/total C ratios (0.20–0.41). There were approximately linear relationships between total C or C_pyr and clay content; two groups of soils gave different linear relationships. A curvilinear relationship between C_pyr and (Al + Fe)_pyr (milli-atom kg⁻¹) that can be approximated by an equation: C_pyr= 53 (Al_pyr+ Fe_pyr)^1/2– 24 was also found for most Thai and Korean soils. The above relationships indicated that total C and C_pyr would be close to zero at zero clay or zero (Al + Fe)_pyr. It was inferred that clay-humus interaction has a primary importance in the determination of humus content in red and yellow soils in tropical and temperate regions and that the main role of clay is to supply Al and Fe that complex and stabilize humus against microbial degradation.     

The topology of pore structure in cracking clay soil I. The estimation of numerical density

The numerical density, N_v , of the pore structure of soil is the number of disjoint networks of pores per unit volume of soil. A method is described for estimating N_v of patterns of cracks that dominate in many clay subsoils. The cracks are photographed from numerous close-spaced parallel sections and skeletonized; by comparing the skeletonized photographs sequentially, individual networks are tracked from one section to another and counted. The average number of networks that appears or disappears per section in the sequence is a measure of the numerical density and is obtained by regressing the counts on the volume of soil spanned by the sections. The regressions for appearances and disappearances converge on one another and stabilize within 10 to 20 sections, so that N_v can be estimated for a sample of soil with moderate effort.
Estimates of N_v for cracks wider than 60 μm in subsoil of the Windsor series, sampled at two nearby sites and 5 years apart in time and determined from sections at 50 μm intervals, were approximately 32 cm⁻³ and 36cm⁻³. That of N_v in the Swanwick series subsoil nearby was about 75 cm⁻³.     

Estimation of the density of the protocatechuate-degrading bacterial community in soil by real-time PCR

The β-ketoadipate pathway is the major route for degradation of aromatic compounds by various soil microorganisms. Protocatechuate 3,4-dioxygenase, a key enzyme of this pathway and which is encoded by pcaGH genes, catalyses the ring cleavage of protocatechuate. Microorganisms harbouring pcaGH genes are widely distributed in the environment but little is known about their relative abundance within the total microflora. Hence, this paper reports the development of a real-time PCR assay to quantify the bacterial pcaH sequence that encodes the β sub-unit of the protocatechuate 3,4-dioxygenase. This real-time PCR assay was linear over seven orders of magnitude with a calculated efficiency of 99% and sensitive up to 10² copies of the pcaH sequence per assay. Real-time PCR analyses performed on six soils with different physico-chemical properties, revealed pcaH densities ranging from 10³ to 10⁴ copies of pcaH  ng⁻¹ of soil DNA, which corresponded to approximately 0.2–10.9% of the total bacterial community. The sequencing of real-time PCR amplicons yielded 48 deduced amino acid sequences that exhibited 44–100% identity to known bacterial PcaH sequences, thereby confirming the accuracy of this real-time PCR assay.     

Effect of supplied phytosiderophore on 59Fe absorption and translocation in Fe-deficient barley grown hydroponically in low phosphorus media

Hydroponically grown barley plants ( Hordeum vulgare L. cv. Minorimugi) under iron-deficient (–Fe) and high phosphorus (P) conditions (500 µmol L⁻¹) showed Fe chlorosis and lower growth compared with plants grown in –Fe and low P conditions (50, 5 and 0.5 µmol L⁻¹). To understand the physiological role of P in regulating the growth of plants in –Fe medium, we carried out an Fe feeding experiment using four P levels (500, 50, 5 and 0.5 µmol L⁻¹) and phytosiderophores (PS), mugineic acid. Our results suggest that plants grown in a high P medium had higher absorption activity of ⁵⁹Fe compared with plants grown in low P media, irrespective of the presence or absence of added PS. Translocation of ⁵⁹Fe from roots to shoots was not affected by the P level. The relative translocation rate of ⁵⁹Fe increased with decreasing levels of P in the medium. In general, the addition of PS enhanced the absorption of ⁵⁹Fe and its translocation. Taken together these results suggest that the lower relative translocation rate of Fe in high P plants may be induced by the physiological inactivation of Fe in the roots, and the higher absorption activity of Fe in high P conditions possibly results from the response of barley plants to Fe deficiency.     

Fixation of radiocaesium traces in a weathering sequence mica → vermiculite → hydroxy interlayered vermiculite

Radiocaesium fixation in soils is reported to occur on frayed edge sites of micaceous minerals. The weathering of mica in acid soils may therefore influence the Cs⁺ fixation process and thereby the mobility of the radiopollutant. We produced a laboratory weathering model biotite → trioctahedral vermiculite → oxidized vermiculite → hydroxy interlayered vermiculite (HIV) and quantified the Cs⁺ fixation of each mineral both in a fixed K⁺–Ca²⁺ background and in acid conditions. The transformation process was achieved through K depletion by Na-tetraphenylboron, oxidation with Br₂ and Al-intercalation using NaOH and AlCl₃. In a constant K⁺–Ca²⁺ background, vermiculite fixed 92–95% of the initial ¹³⁷Cs⁺ contamination while biotite and HIV fixed only 18–33%. In acid conditions, the interlayer occupancy by either potassium (biotite) or hydroxy-Al groups (HIV) strongly limited Cs⁺ fixation to 1–4% of the initial ¹³⁷Cs⁺ contamination. Cs⁺ fixation occurred on vermiculitic sites associated with micaceous wedge zones. Though both oxidized and trioctahedral vermiculites fixed similar Cs⁺ amounts in a constant K⁺–Ca²⁺ background (92–95%), the oxidized vermiculite retained much more radiocaesium in acid conditions (78–84% against 54–59%), because of its dioctahedral character.     

Nitrogen inputs and outputs in a small agricultural catchment in the eastern part of the United Kingdom

Abstract. Nitrate concentrations measured in an ephemeral stream draining a 170 ha clay catchment in eastern England, with about 23% arable land, were greater than 11.3 mg N 1–¹ on the resumption of flow each autumn but then declined. There was also a spring peak in two years out of seven, 1978–1984, which depend on the length of time soils was at field capacity in the preceding winter. Mean annual load measured in rain was 19 kg N ha^-1 and loss of nitrate in the stream 34 kg N ha^-1. A catchment nitrogen balance suggested that inputs, which averaged 130 kg N ha yr^-1, were generally more than outputs, average 108 kg N ha yr^-1', but gaseous losses were not taken into account.