Enzyme activities and microbial biomass in coastal soils of India

Soil salinity is a serious problem for agriculture in coastal regions, wherein salinity is temporal in nature. We studied the effect of salinity, in summer, monsoon and winter seasons, on microbial biomass carbon (MBC) and enzyme activities (EAs) of the salt-affected soils of the coastal region of the Bay of Bengal, Sundarbans, India. The average pH of soils collected from different sites, during different seasons varied from 4.8 to 7.8. The average organic C (OC) and total N (TN) content of the soils ranged between 5.2-14.1 and 0.6-1.4 g kg⁻¹, respectively. The electrical conductivity of the saturation extract (ECe) of soils, averaged over season, varied from 2.2 to 16.3 dSm⁻¹. The ECe of the soils increased five fold during the summer season (13.8 dSm⁻¹) than the monsoon season (2.7 dSm⁻¹). The major cation and anion detected were Na⁺ and Cl⁻, respectively. Seasonality exerted considerable effects on MBC and soil EAs, with the lowest values recorded during the summer season. The activities of β-glucosidase, urease, acid phosphatase and alkaline phosphatase were similar during the winter and monsoon season. The dehydrogenase activity of soils was higher in monsoon than in winter. Average MBC, dehydrogenase, β-glucosidase, urease, acid phosphatase and alkaline phosphatase activities of the saline soils ranged from 125 to 346 mg kg⁻¹ oven dry soil, 6-9.9 mg triphenyl formazan (TPF) kg⁻¹ oven dry soil h⁻¹, 18-53 mg p-nitro phenol (PNP) kg⁻¹ oven dry soil h⁻¹, 38-86 mg urea hydrolyzed kg⁻¹ oven dry soil h⁻¹, 213-584 mg PNP kg⁻¹ oven dry soil h⁻¹ and 176-362 mg PNP g⁻¹ oven dry soil h⁻¹, respectively. The same for the non-saline soils were 274-446 mg kg⁻¹ oven dry soil, 8.8-14.4 mg TPF kg⁻¹ oven dry soil h⁻¹, 41-80 mg PNP kg⁻¹ oven dry soil h⁻¹, 89-134 mg urea hydrolyzed kg⁻¹ oven dry soil h⁻¹, 219-287 mg PNP kg⁻¹ oven dry soil h⁻¹ and 407-417 mg PNP kg⁻¹ oven dry soil h⁻¹, respectively. About 48%, 82%, 48%, 63%, 40% and 48% variation in MBC, dehydrogenase activity, β-glucosidase activity, urease activity, acid phosphatase activity and alkaline phosphatase activity, respectively, could be explained by the variation in ECe of saline soils. Suppression of EAs of the coastal soils during summer due to salinity rise is of immense agronomic significance and needs suitable interventions for sustainable crop production.     

Measuring rates of gross and net mineralisation of organic phosphorus in soils

The isotopic dilution method developed by Oehl et al. [2001b. Organic phosphorus mineralisation studies using isotopic dilution techniques. Soil Science Society of America Journal 65, 780-787] to measure gross mineralisation of soil organic phosphorus (P) was tested on a range of low-P sorbing soils. This isotopic dilution method relies on accurate prediction of radiolabel behaviour due to soil physicochemical processes. Based on experimental validation of the extrapolation for isotopic dilution due to physicochemical processes using autoclaved soils, a simple power function was used for extrapolation rather than the more complex equation used in the original method. For several soils, however, a potential overestimation of gross mineralisation by 0.1-2.0 mg P kg⁻¹ d⁻¹ was revealed. In addition, the detection limit of P mineralisation ranged between 0.6 and 2.6 mg P kg⁻¹ d⁻¹. The method is likely to be at the detection limit for soils that are high in available P and low in biological activity. The method was modified with respect to the extrapolation and successfully applied to a soil with relatively high microbial P (18 mg P kg⁻¹) and soil respiration rates (29 mg C kg⁻¹ d⁻¹), revealing gross mineralisation rates of organic P of 0.9-1.2 mg P kg⁻¹ d⁻¹. Measurement of uptake of ³²P by the microbial biomass allowed derivation of a net organic P mineralisation rate of 0.5-0.9 mg P kg⁻¹ d⁻¹.     

Cyanobacterial inoculation of heated soils: effect on microorganisms of C and N cycles and on chemical composition in soil surface

Physiological groups of soil microorganisms, total C and N and available nutrients were investigated in four heated (350 °C, 1 h) soils (one Ortic Podsol over sandstone and three Humic Cambisol over granite, schist or limestone) inoculated (1.5 μg chlorophyll a g⁻¹ soil or 3.0 μg chlorophyll a g⁻¹ soil) with four cyanobacterial strains of the genus Oscillatoria, Nostoc or Scytonema and a mixture of them.Cyanobacterial inoculation promoted the formation of microbiotic crusts which contained a relatively high number of NH₄⁺-producers (7.4×10⁹ g⁻¹ crust), starch-mineralizing microbes (1.7×10⁸ g⁻¹ crust), cellulose-mineralizing microbes (1.4×10⁶ g⁻¹ crust) and NO₂⁻ and NO₃⁻ producers (6.9×10⁴ and 7.3×10³ g⁻¹ crust, respectively). These crusts showed a wide range of C and N contents with an average of 293 g C kg⁻¹ crust and 50 g N kg⁻¹ crust, respectively. In general, Ca was the most abundant available nutrient (804 mg kg⁻¹ crust), followed by Mg (269 mg kg⁻¹ crust), K (173 mg kg⁻¹ crust), Na (164 mg kg⁻¹ crust) and P (129 mg kg⁻¹ crust). There were close positive correlations among all the biotic and abiotic components of the crusts.Biofertilization with cyanobacteria induced great microbial proliferation as well as high increases in organic matter and nutrients in the surface of the heated soils. In general, cellulolytics were increased by four logarithmic units, amylolytics and ammonifiers by three logarithmic units and nitrifiers by more than two logarithmic units. C and N contents rose an average of 275 g C kg⁻¹ soil and 50 g N kg⁻¹ soil while the C:N ratio decreased up to 7 units. Among the available nutrients the highest increase was for Ca (315 mg kg⁻¹ soil) followed by Mg (189 mg kg⁻¹ soil), K (111 mg kg⁻¹ soil), Na (109 mg kg⁻¹ soil) and P (89 mg kg⁻¹ soil). Fluctuations of the microbial groups as well as those of organic matter and nutrients were positively correlated.The efficacy of inoculation depended on both the type of soil and the class of inoculum. The best treatment was the mixture of the four strains and, whatever the inoculum used, the soil over lime showed the most developed crust followed by the soils over schist, granite and sandstone. In the medium term there were not significant differences between the two inocula amounts tested.These results showed that inoculation of burned soils with alien N₂-fixing cyanobacteria may be a biotechnological means of promoting microbiotic crust formation, enhancing C and N cycling microorganisms and increasing organic matter and nutrient contents in heated soils.     

Basal organic phosphorus mineralization in soils under different farming systems

Soil organic P (P_o) mineralization plays an important role in soil P cycling. Quantitative information on the release of available inorganic P (P_i) by this process is difficult to obtain because any mineralized P_i gets rapidly sorbed. We applied a new approach to quantify basal soil P_o mineralization, based on ³³PO₄ isotopic dilution during 10 days of incubation, in soils differing in microbiological activity. The soils originated from a 20 years old field experiment, including a conventional system receiving exclusively mineral fertilizers (MIN), a bio-organic (ORG) and bio-dynamic (DYN) system. Indicators of soil microbiological activity, such as size and activity of the soil microbial biomass and phosphatase activity, were highest in DYN and lowest in MIN. In order to assess P_o hydrolysis driven by phosphatase in sterile soils, a set of soil samples was γ-irradiated. Basal P_o mineralization rates in non-irradiated samples were between 1.4 and 2.5 mg P kg⁻¹ day⁻¹ and decreased in the order DYN>ORG≥MIN. This is an amount lower, approximately equivalent to, or higher than water soluble P_i of MIN, ORG and DYN soils, respectively, but in every soil was less than 10% of the amount of P isotopically exchangeable during one day. This shows that physico-chemical processes are more important than basal mineralization in releasing plant available P_i. Organic P mineralization rates were higher, and differences between soils were more pronounced in γ-irradiated than in non-irradiated soils, with mineralization rates ranging from 2.2 to 4.6 mg P kg⁻¹ day⁻¹. These rates of hydrolysis, however, cannot be compared to those in non-sterile soils as they are affected by the release of cellular compounds, e.g. easily mineralizable P_o, derived from microbial cells killed by γ-irradiation.     

Unusually high levels of bio-available phosphate in the soils of Ogasawara Islands, Japan: Putative influence of seabirds

Ogasawara Islands are important ecosystems sustaining many indigenous spices. To clarify the indigenous soil environments of Ogasawara Islands, we studied the chemistry of the soils. Many surface soils were low in bio-available P (0 to 0.55 g P₂O₅ kg⁻¹, average: 0.04 g P₂O₅ kg⁻¹ as Bray II P, n = 22), but several soils were found to contain extremely large amounts of bio-available P (1.36 to 6.98 g P₂O₅ kg⁻¹, average: 2.93 g P₂O₅ kg⁻¹, n = 5). From soil profile analyses, the authors concluded that the extremely large amount of bio-available P could not be explained by the effects of parent materials with high P contents nor the effect of fertilizations by human activity, but the effects of natural seabird activities in the past could be the cause. The soil profiles with large amounts of bio-available P indicate deep migration of soil materials from A horizons, which could be a result of intensive mixing of upper horizons by seabird activities. The intensive mixing was supported by the low mechanical impedance of the horizons for the P-accumulating soils (8.17 ± 2.54 kg cm⁻², n = 8) than those for the non-P-accumulating soils (17.46 ± 3.52 kg cm⁻², n = 36). It is likely that in the past seabirds, such as shearwaters, made burrows in the soils for nesting and propagating and inadvertently transported a large amount of P from the sea to the soils, resulting in the extremely large amounts of bio-available P in the present soils.     

Zinc toxicity on N2O reduction declines with time in laboratory spiked soils and is undetectable in field contaminated soils

Denitrification assays in soils spiked with zinc salt have shown inhibition of the N₂O reduction resulting in increased soil N₂O fluxes with increasing soil Zn concentration. It is unclear if the same is true for environmentally contaminated soils. Net production of N₂O and N₂ was monitored during anaerobic incubations (25 °C, He atmosphere) of soils freshly spiked with ZnCl₂ and of corresponding soils that were gradually enriched with metals (mainly Zn) in the field by previous sludge amendments or by corrosion of galvanized structures. Total denitrification activity (i.e. the sum of N₂O+N₂ production rate) was not inhibited by freshly added Zn salts up to 1600 mg Zn kg⁻¹, whereas N₂O reduction decreased by 50% (EC₅₀) at total Zn concentrations of 231 mg Zn kg⁻¹ (ZEV soil) and 368 mg Zn kg⁻¹ (TM soil). In contrast, N₂O reduction was not reduced by soil Zn in any of the field contaminated soils, even at total soil Zn or soil solution Zn concentrations exceeding more than 5 times corresponding EC₅₀'s of the freshly spiked soil. The absence of adverse effects in the field contaminated soils was unrelated to soil NO₃ or organic matter concentration. Ageing (2-8 weeks) and soil leaching after spiking reduced the toxicity of Zn on N₂O reduction, either expressed as total Zn or soil solution Zn, suggesting adaptation reactions. However, no full recovery after spiking was identified at the largest incubation period in one soil. In addition, the denitrification assay performed with sewage sludge showed elevated N₂O release in Zn contaminated sludges (>6000 mg Zn kg⁻¹ dry matter) whereas this was not observed in low Zn sludge (<1000 mg Zn kg⁻¹ dry matter) suggesting limits to adaptation reactions in the sludge particles. It is concluded that the use of soils spiked with Zn salts overestimates effects on N₂O reduction. Field data on N₂O fluxes in sludge amended soils are required to identify if metals indeed promote N₂O emissions in sludge amended soils.     

Heavy metal accumulation by two earthworm species and its relationship to total and DTPA-extractable metals in soils

The concentrations of Zn, Cd, Pb and Cu in earthworm tissues were compared with the total and DTPA-extractable contents of these heavy metals in contaminated soils. Samples were taken from a pasture polluted by waste from a metallurgic industry over 70 y ago. Three individuals of Aporrectodea caliginosa and Lumbricus rubellus and soil samples were collected at six points along a gradient of increasing pollution. Total metal contents of earthworms, soil, and metals extracted by DTPA from the soil were measured. Total heavy metal contents of the soils ranged from 165.7 to 1231.7 mg Zn kg⁻¹, 2.7 to 5.2 mg Cd kg⁻¹, 45.8 to 465.5 mg Pb kg⁻¹ and 30.0 to 107.5 mg Cu kg⁻¹. Their correlations with metals extracted by DTPA were highly significant. Contents of the metals in earthworm tissues were higher in A. caliginosa than in L. rubellus, with values ranging from 556 to 3381 mg Zn kg⁻¹, 11.6 to 102.9 mg Cd kg⁻¹, 1.9 to 182.8 mg Pb kg⁻¹ and 17.9 to 35.9 mg Cu kg⁻¹ in A. caliginosa, and from 667.9 to 2645 mg Zn kg⁻¹, 7.7 to 26.3 mg Cd kg⁻¹, 0.5 to 37.9 mg Pb kg⁻¹ and 16.0 to 37.6 mg Cu kg⁻¹ in L. rubellus, respectively. Correlations between body loads in earthworms with either total or DTPA-extractable contents of soil metals were significant, except for Cd in L. rubellus and Cu in A. caliginosa. Considering its simple analytical procedure, DTPA-extractable fraction may be preferable to total metal content as a predictor of bio-concentrations of heavy metals in earthworms. Biota-to-Soil Accumulation Factor (BSAF) of these four metals are Cd>Zn>Cu>Pb, with range of mean values between: Cd (6.18-17.02), Zn (1.95-7.91), Cu (0.27-0.89) and Pb (0.08-0.38) in A. caliginosa, and Cd (3.64-6.34), Zn (1.5-6.35), Cu (0.29-0.87) and Pb (0.04-0.13) in L. rubellus. The BSAF of Ca, Fe and Mn are Ca>Mn>Fe, with mean values of: Ca (0.46-1.31), Mn (0.041-0.111), Fe (0.017-0.07) in A. caliginosa and Ca (0.98-2.13), Mn (0.14-0.23), Fe (0.019-0.048) in L. rubellus, respectively. Results of principal component analysis showed that the two earthworm species differ in the pattern of metal bioaccumulation which is related to their ecological roles in contaminated soils.     

Influence of selenium on oxidoreductive enzymes activity in soil and in plants

The aim of this greenhouse experiment was the assessment of the influence of H₂SeO₃ at soil concentrations of 0.05, 0.15 and 0.45 mmol kg⁻¹, on the activity of selected oxidoreductive enzymes in wheat (Triticum aestivum). The wheat plants were grown in 2 dm³ pots filled with dust-silt black soil of pH 7.7. Applied H₂SeO₃ caused activation of plant nitrate reductase at all concentrations, but activation of plant polyphenol oxidase at only two lower concentrations. The highest concentration caused inhibition of polyphenol oxidase and peroxidase. Plant catalase activity decreased under the influence of 0.15 and 0.45 mmol kg⁻¹ concentration. After the final analysis Se was quantified in plants and soil. The amounts in plants were: control (unamended soil) 1.95 mg kg⁻¹; I dose (0.05 mmol kg⁻¹) 18.27 mg kg⁻¹; II dose (0.15 mmol kg⁻¹) 33.20 mg kg⁻¹ and III dose (0.45 mmol kg⁻¹) 38.37 mg kg⁻¹, in soil: 0.265 mg kg⁻¹; 3.61 mg kg⁻¹; 10.53 mg kg⁻¹; 30.53 mg kg⁻¹; respectively. Simultaneously, a laboratory experiment was performed, where the activity of soil catalase and peroxidase were tested after 1, 3, 7, 14, 28, 56, and 112 days after Se treatment. Peroxidase activity in soil decreased with increasing Se content, over the whole experiment. The lowest dose of Se caused activation a significant 10% increase in catalase activity, but the influence of others doses was unclear.     

Surface runoff phosphorus (P) loss in relation to phosphatase activity and soil P fractions in Florida sandy soils under citrus production

Phosphorus losses by surface runoff from agricultural lands have been of public concern due to increasing P contamination to surface waters. Five representative commercial citrus groves (C1-C5) located in South Florida were studied to evaluate the relationships between P fractions in soils, surface runoff P, and soil phosphatase activity. A modified Hedley P sequential fractionation procedure was employed to fractionate soil P. Soil P consisted of mainly organically- and Ca/Mg-bound P fractions. The organically-bound P (biological P, sum of organic P in the water, NaHCO₃ and NaOH extracts) was dominant in the acidic sandy soils from the C2 and C3 sites (18% and 24% of total soil P), whereas the Ca/Mg-bound P (HCl-extractable P) accounted for 45-60% of soil total P in the neutral and alkaline soils (C1, C4 and C5 soils). Plant-available P (sum of water and NaHCO₃ extractable P fractions) ranged from 27 to 61 mg P kg⁻¹ and decreased in the order of C3>C4>C1>C2>C5. The mean total P concentrations (TP) in surface runoff water samples ranged from 0.51 to 2.64 mg L⁻¹. Total P, total dissolved P (TDP), and PO₄³⁻-P in surface runoff were significantly correlated with soil biological P and plant-available P forms (p<0.01), suggesting that surface runoff P was directly derived from soil available P pools, including H₂O- and NaHCO₃- extractable inorganic P, water-soluble organic P, and NaHCO₃- and NaOH-extractable organic P fractions, which are readily mineralized by soil microorganisms and/or enzyme mediated processes. Soil neutral (55-190 mg phenol kg⁻¹ 3 h⁻¹) and natural (measured at soil pH) phosphatase activities (77-295 mg phenol kg⁻¹ 3 h⁻¹) were related to TP, TDP, and PO₄³⁻-P in surface runoff, and plant-available P and biological P forms in soils. These results indicate that there is a potential relationship between soil P availability and phosphatase activities, relating to P loss by surface runoff. Therefore, the neutral and natural phosphatase activities, especially the natural phosphatase activity, may serve as an index of surface runoff P loss potential and soil P availability.     

Toxicity of imidacloprid to the earthworm Eisenia andrei and collembolan Folsomia candida in three contrasting tropical soils

Purpose

Imidacloprid is a widely used seed dressing insecticide in Brazil. However, the effects of this pesticide on non-target organisms such as soil fauna still present some knowledge gaps in tropical soils. This study aimed to assess the toxicity and risk of imidacloprid to earthworms Eisenia andrei and collembolans Folsomia candida in three contrasting Brazilian tropical soils.

Materials and methods

Acute and chronic toxicity assays were performed in the laboratory with both species in a tropical artificial soil (TAS) and in two natural soils (Oxisol and Entisol), at room temperature of 25 °C. The ecological risk was calculated for each species and soil by using the toxicity exposure ratio (TER) and hazard quotient (HQ) approaches.

Results and discussion

Acute toxicity for collembolans and earthworms was higher in Entisol (LC₅₀?=?4.68 and 0.55 mg kg^?1, respectively) when compared with TAS (LC₅₀?=?10.8 and 9.18 mg kg^?1, respectively) and Oxisol (LC_{50collembolans}?=?25.1 mg kg^?1). Chronic toxicity for collembolans was similar in TAS and Oxisol (EC_{50 TAS}?=?0.80 mg kg^?1; EC_{50 OXISOL}?=?0.83 mg kg^?1), whereas higher toxicity was observed in Entisol (EC₅₀?=?0.09 mg kg^?1). In chronic assays with earthworms, imidacloprid was also more toxic in Entisol (EC₅₀?=?0.21 mg kg^?1) when compared to TAS (EC₅₀?=?1.89 mg kg^?1). TER and HQ values indicated a significant risk of exposure of the species to imidacloprid in all soils tested, and the risk in Entisol was at least six times higher than in Oxisol or TAS.

Conclusions

The toxicity and risk of imidacloprid varied significantly between tropical soils, being the species exposure to this pesticide particularly hazardous in very sandy natural soils such as Entisol.

     

Assessing the toxic effect of nitrapyrin on nitrogen transformation in soil

A 28 d N transformation test was developed according to the OECD guideline 216. In the laboratory-based test, a suitable soil was amended with powdered plant meal as an organic N source. Soil samples of 1 kg treated with five concentrations of nitrapyrin (2-chloro-6-(trichloromethyl)-pyridine), in the range 1.0-100 mg kg⁻¹ dry weight were incubated for 28 d at 20±2 °C. A dose response was produced and the N mineralisation EC₅₀ (95% C.I.) for nitrapyrin was 3.1 (1.9-4.3) mg kg⁻¹ dry soil. The determined EC₅₀ was compared with literature figures for similar end points but using different methodology.     

Possibilities to reduce rice straw-induced global warming potential of a sandy paddy soil by combining hydrological manipulations and urea-N fertilizations

Global warming potential (GWP) of sandy paddy soils may be reduced by trade-offs between N₂O, CH₄ and CO₂ emissions. Laboratory experiments using either rice straw (1% or 0.5%) or together with urea-N (25 or 50 mg N kg⁻¹ soil) at various levels of soil water were carried out for 30 days each, to test this assumption. Waterlogging combined with urea-N increased total N₂O emissions, with greater release upon rewaterlogging (7.4 mg N kg⁻¹ soil) than experienced by removing waterlogging only. Rice straw±urea-N either emitted small amounts of N₂O or resulted in negative values at all water levels, including saturated and aerobic. Total CH₄ fluxes declined with the decreased water levels and amount of rice straw (<193 mg C kg⁻¹ soil), and also for CO₂ with the latter (<1340 mg C kg⁻¹ soil), and rewaterlogging had little influence on both. N₂O under rewaterlogged and waterlogged±urea-N, CH₄ under waterlogged with rice straw, and CO₂ for the remainder were the major contributors to GWP. Results show that waterlogging following aerobic decomposition of rice straw (1%) with urea-N, applied either at the beginning or at the end of the aerobic conditions, could decrease GWP by 56-64% and 32-42% over the sole addition of rice straw (1% and 0.5%) under waterlogged and saturated conditions, respectively.     

Increased degradation of phenanthrene in soil by Pseudomonas sp. GF3 in the presence of wheat

A phenanthrene-degrading bacterial strain Pseudomonas sp. GF3 was examined for plant-growth promoting effects and phenanthrene removal in soil artificially contaminated with low and high levels of phenanthrene (0, 100 and 200 mg kg⁻¹) in pot experiments. Low and high phenanthrene treatments significantly decreased the growth of wheat. Inoculation with bacterial strain Pseudomonas sp. GF3 was found to increase root and shoot growth of wheat. Strain GF3 was able to degrade phenanthrene effectively in the unplanted and planted soils. Over a period of 80 days the concentration of phenanthrene in soil in which wheat was grown was significantly lower than in unplanted soil (p<0.05). At the end of the 80-d experiments, 62.2% and 42.3% of phenanthrene had disappeared from planted soils without Pseudomonas sp. GF3 when the phenanthrene was added at 100 and 200 mg kg⁻¹ soil, respectively, but 84.8% and 70.2% of phenanthrene had disappeared from planted soils with the bacterial inoculation. The presence of vegetation significantly enhances the dissipation of phenanthrene in the soil. There was no significant difference in soil polyphenol oxidase activities among the applications of 0, 100 and 200 mg kg⁻¹ of phenanthrene. However, the enzyme activities in planted and unplanted soils inoculated with the strain Pseudomonas sp. GF3 were significantly higher than those of non-inoculation controls. The bacterial isolate was also able to colonize and develop in the rhizosphere soil of wheat after inoculation.     

Immobilization and remineralization of N following addition of wheat straw into soil: determination of gross N transformation rates by N-ammonium isotope dilution technique

N dynamics in soil where wheat straw was incorporated were investigated by a soil incubation experiment using ¹⁵N-labelled nitrate or ¹⁵N-labelled wheat straw. The incubated soils were sampled after 7, 28, 54 days from the incorporation of wheat straw, respectively, and gross rates of N transformations including N remineralization and temporal changes in the amount of microbial biomass were determined.Following the addition of wheat straw into soils, rapid decrease of nitrate content in soil and increase of microbial biomass C and N occurred within the first week from onset of the experiment. Both the gross rates of mineralization and immobilization determined by ¹⁵N-ammonium isotope dilution technique were remarkably enhanced by the addition of wheat straw, and gradually decreased with time. Remineralization rate of N derived from ¹⁵N-labelled nitrate, and mineralization rate of N derived from ¹⁵N-labelled wheat straw was estimated by ¹⁵N isotope dilution technique using non-labelled ammonium. Remineralization rates of N derived from ¹⁵N-labelled nitrate were calculated to be 0.71 mg N kg⁻¹ d⁻¹ after 7 days, 0.55 mg N kg⁻¹ d⁻¹ after 28 days, and 0.29 mg N kg⁻¹ d⁻¹ after 54 days.Nearly 10% of the ¹⁵N-labelled N originally contained in the wheat straw was held in the microbial biomass irrespective of the sampling time. The amount of inorganic N in soil which was derived from ¹⁵N-labelled wheat straw ranged between 1.93 and 2.37 mg N kg⁻¹.Rates of N transformations in soil with ¹⁵N-labelled wheat straw were obtained by assuming that the k value was equal to the ¹⁵N abundance of biomass N, and the obtained values were considered to be valid.     

Water dispersible clay in calcareous soils of southwestern Spain

Water dispersible clay (WDC) is a good indicator of the risk of soil erosion by water and the consequent losses of nutrients and contaminants in overland flow. We measured the content and studied the properties of WDC in 26 samples of calcareous Xeralfs, Xerepts and Xererts of southwestern Spain collected from fields under different crop and tillage management; the soils ranged widely in total clay content (60–455 g kg^− 1), calcium carbonate equivalent (CCE) (< 1–559 g kg^− 1) and active calcium carbonate equivalent (ACCE; 2–135 g kg^− 1), and were poor in organic carbon and soluble salts. The WDC content was determined by shaking 10 g of soil in 1 L of simulated rainwater for 4 hours. Non-carbonate WDC contents were found to be strongly correlated with the total clay content of the soils and ranged from 1 to 92 g kg^− 1 soil (mean = 29 g kg^− 1), the non-carbonate WDC/total clay ratio ranging from 0.01 to 0.29 (mean = 0.12). Based on regression analyses, illite was more dispersible than smectite and iron oxides decreased dispersion of clay. Carbonate WDC contents ranged from 1 to 27 g kg^− 1 (mean = 8 g kg^− 1) and were averaged one third the non-carbonate WDC contents; also, they were strongly correlated with the soil ACCE. WDC was rich in phosphorus (P) relative to the bulk soil. The enrichment ratio (ER) for total P (i.e. the ratio of total P in WDC to total P in soil) ranged from 0.2 to 29 (mean = 5) and was inversely related to the total clay content. On average, about one tenth of the soil total P was exported in the WDC and about one fifth of the total P in WDC was in the form of bicarbonate-extractable P (i.e. relatively soluble or ‘labile’ P). Part of the P in WDC seemingly occurred as metal phosphate particles formed by reaction of P fertilizers with soil. In summary, significant amounts of P can be exported via WDC, even though the proportion of total clay that is water dispersible is substantially lower in these soils than in cultivated soils of other semiarid regions.     

Response of microbial activity and microbial community composition in soils to long-term arsenic and cadmium exposure

Arsenic (As) and cadmium (Cd) in soils can affect soil microbial function and community composition and, therefore, may have effects on soil ecosystem functioning. The aim of our study was to assess the effects of long-term As and Cd contamination on soil microbial community composition and soil enzyme activities. We analyzed soils that have been contaminated 25 years ago and at present still show enhanced levels of either As, 18 and 39 mg kg⁻¹, or Cd, 34 and 134 mg kg⁻¹. Soil without heavy metal addition served as control. Polymerase chain reaction (PCR) followed by denaturing gradient gel electrophoresis (DGGE) showed that bacterial community composition in As and Cd contaminated soils differed from that in the control soil. The same was true for the microbial community composition assessed by analysis of respiratory quinones. Soil fungi and Proteobacteria appeared to be tolerant towards As and Cd, while other groups of bacteria were reduced. The decline in alkaline phosphatase, arylsulphatase, protease and urease activities in the As- and Cd-contaminated soils was correlated with a decrease of respiratory quinones occuring in Actinobacteria and Firmicutes. Xylanase activity was unaffected or elevated in the contaminated soils which was correlated with a higher abundance of fungal quinones, and quinones found in Proteobacteria.     

Transfer and loss of naturally-occurring plasmids among isolates of Rhizobium leguminosarum bv. viciae in heavy metal contaminated soils

Plasmid transfer among isolates of Rhizobium leguminosarum bv. viciae in heavy metal contaminated soils from a long-term experiment in Braunschweig, Germany, was investigated under laboratory conditions. Three replicate samples each of four sterilized soils with total Zn contents of 54, 104, 208 and 340 mg kg⁻¹ were inoculated with an equal number (1×10⁵ cells g⁻¹ soil) of seven different, well-characterized isolates of R. leguminosarum bv. viciae. Four of the isolates were from an uncontaminated control plot (total Zn 54 mg kg⁻¹) and three were from a metal-contaminated plot (total Zn 340 mg kg⁻¹).After 1 year the population size was between 10⁶ and 10⁷ g⁻¹ soil, and remained at this level in all but the most contaminated soil. In the soil from the most contaminated plot no initial increase in rhizobial numbers was seen, and the population declined after 1 year to <30 cells g⁻¹ soil after 4 years. One isolate originally from uncontaminated soil that had five large plasmids (no. 2-8-27) was the most abundant type re-isolated from all of the soils. Isolates originally from the metal-contaminated soils were only recovered in the most contaminated soil. After 1 year, four isolates with plasmid profiles distinct from those inoculated into the soils were recovered. One isolate in the control soil appeared to have lost a plasmid. Three isolates from heavy metal contaminated soils (one isolate from the soil with total Zn 208 mg kg⁻¹ and two isolates from the soil with total Zn 340 mg kg⁻¹) had all acquired one plasmid. Plasmid transfer was confirmed using the distinct ITS-RFLP types of the isolates and DNA hybridization using probes specific to the transferred plasmid. The transconjugant of 2-8-27 which had gained a plasmid was found in one replicate after 2 years of the most contaminated soil but comprised more than 50% of the isolates. A similar type appeared in a separate replicate of the most contaminated soil after 3 years and persisted in both of these soils until the final sampling after 4 years. After 2 years isolates were recovered from four of the soil replicates with the chromosomal type of 2-8-27 which appeared to have lost one plasmid, but these were not recovered subsequently.Isolate 2-8-27 was among the isolates most sensitive to Zn in laboratory assays, whereas isolate 7-13-1 showed greater zinc tolerance. Acquisition of the plasmid conferred enhanced Zn tolerance to the recipients, but transconjugant isolates were not as metal tolerant as 7-13-1, the putative donor. Laboratory matings between 2-8-27 and 7-13-1 in the presence of Zn resulted in the conjugal transfer of the same small plasmid from 7-13-1 to isolate 2-8-27 and the transconjugant had enhanced metal tolerance. Our results show that transfer of naturally-occurring plasmids among rhizobial strains is stimulated by increased metal concentrations in soil. We further demonstrate that the transfer of naturally-occurring plasmids is important in conferring enhanced tolerance to elevated zinc concentrations in rhizobia.     

Denitrification enzyme activity and potential of subsoils under grazed grasslands assayed by membrane inlet mass spectrometer

The importance of subsoil denitrification on the fate of agriculturally derived nitrate (NO₃) leached to groundwater is crucial for budgeting N in an ecosystem and for identifying areas where the risk of excess NO₃ is reduced. However, the high atmospheric background of di-nitrogen (N₂) causes difficulties in assessing denitrification enzyme activity (DEA) and denitrification potential (DP) in soils directly. Here, we apply Membrane Inlet Mass Spectrometry (MIMS) technique to investigate indirectly DEA and DP in soils by measuring N₂/Ar ratio changes in headspace water over soil. Soils were collected from 0-10, 15-25 and 60-70 cm depths of a grazed ryegrass and grass-clover. The samples were amended with helium-flushed deionized water containing ranges of NO₃ and carbon (glucose-C) and were incubated for six hours in the dark at 21 °C. The peaks for N₂/Ar ratio, declined with increasing soil depth, indicating a reduced substrate requirements to initiate DEA en-masse (15-30 mg NO₃-N alone or with 60-120 mg glucose-C, kg⁻¹ soil). The dissolved N₂O concentrations were very small (0.004-0.269 μg N kg⁻¹ soil) but responded well to the added N and C, showing a reduction in DEA with soil depth. In three separate studies, only subsoils were incubated for 3 days at 12 °C with 20-30 mg NO₃-N ± 40-60 mg glucose-C, kg⁻¹ soil. Denitrification capacity (DC, NO₃ only treatment) was not statistically different to the control (no amendment) within a land use (0.03-0.05 vs. 0.07-0.22 mg N kg⁻¹ soil d⁻¹), the highest being in ryegrass subsoils receiving groundwater. The DP was significantly (P < 0.0001) higher in subsoils under ryegrass than under grass-clover (0.50-0.71 vs. 1.15 mg N kg⁻¹ soil d⁻¹). The rates of DP (NO₃ + glucose-C) increased significantly (P < 0.0001) in unsaturated and saturated subsoils (0.92 and 2.19 mg N kg⁻¹ soil d⁻¹, respectively) of grass-clover, due to the higher reductive state resulting from the 10 day pre-incubation. Available C accelerated denitrification in soils and superseded the temporary elevation in oxidative state due to NO₃ addition. The substrates load differences between the land uses regulated the degree of denitrification rates. Results suggest that both dissolved N₂O measured by gas chromatography and N₂/Ar ratio measured by MIMS to indirectly determine DEA, and the latter to quantify total DC/DP in soils can be used. However, interference of oxygen in the MIMS system should be considered if available C is added or is naturally elevated in soil or groundwater.     

Land use effects on gross nitrogen mineralization, nitrification, and N2O emissions in ephemeral wetlands

Stable ¹⁵N isotope dilution and tracer techniques were used in cultivated (C) and uncultivated (U) ephemeral wetlands in central Saskatchewan, Canada to: (1) quantify gross mineralization and nitrification rates and (2) estimate the relative proportion of N₂O emissions from these wetlands that could be attributed to denitrification versus nitrification-related processes. In-field incubation experiments were repeated in early May, mid-June and late July. Mean gross mineralization and nitrification rates (10.3 and 3.1 mg kg⁻¹ d⁻¹, respectively) did not differ between C and U wetlands on any given date. Despite these similarities, the mean NH₄⁺ pool size in the U wetlands (17.2 mg kg⁻¹) was two to three times that of the C wetlands (6.7 mg kg⁻¹) whereas the mean NO₃⁻ pool size in U wetlands (2.2 mg kg⁻¹) was less than half that of C wetlands (5.8 mg kg⁻¹). Mean N₂O emissions from the C wetlands decreased from 112.8 to 17.0 ng N₂O m² s⁻¹ from May to July, whereas mean U-wetland N₂O emissions ranged only from 31.8 to 51.1 ng N₂O m² s⁻¹ over the same period. This trend is correlated to water-filled pore space in C wetlands, demonstrating a soil moisture influence on emissions. Denitrification is generally considered the dominant emitter of N₂O under anaerobic conditions, but in the C wetlands, only 49% of the May emissions could be directly attributed to denitrification, decreasing to 29% in July. In contrast, more than 75% of the N₂O emissions from the U wetlands arose from denitrification of the soil NO₃⁻ pool throughout the season. These land use differences in emission sources and rates should be taken into consideration when planning management strategies for greenhouse gas mitigation.     

Black carbon in a temperate mixed-grass savanna

Black carbon (BC) or charcoal is thought to represent an important component of the carbon cycle, but has seldom been quantified in soils. We quantified soil BC in a temperate mixed-grass savanna in the southern Great Plains using benzenecarboxylic acids as molecular markers for BC. Soils were collected from four fire treatments (repeated summer fires in 1992 and 1994; repeated winter fires in 1991, 1993 and 1995; alternate-season fires in winter 1991, summer 1992, and winter 1994; and unburned control) at 0-10 and 10-20 cm depth in 1996. Black carbon concentrations ranged from 50 to 130 g BC kg⁻¹ of soil organic carbon (SOC), or from 0.55 to 1.07 g BC kg⁻¹ of whole soil in this mixed grass savanna. The BC contribution to SOC increased significantly with soil depth (P<0.05). Repeated fires increased BC slightly compared to the unburned controls; however, the effects of repeated fires on BC were not statistically significant in this mixed-grass savanna. Results of this study provide estimates of BC concentrations for native, uncultivated mixed-grass savanna, and indicate that 2-3 fires have little effect on the size of the soil BC pool in this region.