Microbial biomass in a semi arid soil of the central highlands of Mexico cultivated with maize or under natural vegetation

Microbial biomass (MB) is the key factor in nutrient dynamics in soil, but no information exists how clearing of vegetation to cultivate maize in the central highlands of Mexico might affect it. Soil MB was measured with the chloroform fumigation incubation (CFI) and fumigation extraction (CFE) techniques and the substrate-induced respiration (SIR) method in soil sampled under or outside the canopy of mesquite (Prosopis laevigata) and huisache (Acacia tortuoso), N₂ fixing shrubs, and from fields cultivated with maize. Microbial biomass C as measured with the CFI technique ranged from 122 mg C kg⁻¹ in agricultural soil to 373 mg C kg⁻¹ in soil sampled under mesquite shrubs. Microbial biomass N as measured with the CFI technique ranged from 11 mg N kg⁻¹ in agricultural soil to 116 mg N kg⁻¹ in soil sampled under mesquite shrub. The ratio of microbial biomass C as measured with CFI related to the ninhydrin-positive compounds (NPC) was 12.23 after 1 day and 8.43 after 10 days while the relationship with extractable C was 3.15 and 2.96, respectively. The metabolic quotient (qCO₂) decreased in the order OUTSIDE > MESQUITE > HUIZACHE > AGRICULTURE, and the microbial biomass:soil organic C ratio decreased in the order MESQUITE > HUIZACHE > OUTSIDE > AGRICULTURE using SIR to determine the microbial biomass. It was found that converting soil under natural vegetation to arable soil was not only detrimental for soil quality, but might be unsustainable as organic matter input is limited.     

Is nitrate reduction to nitrite possible in glucose-amended alkaline saline soil under aerobic conditions?

A phylogenetic analysis of the archaeal community in the soil of the former Lake Texcoco showed that some of the clones identified were affiliated to Archeae that reduce nitrate (NO₃^?) to nitrite (NO₂^?) and NO₂^? to unknown products under aerobic conditions. Previous research suggested that this indeed might occur when an easily decomposable C-substrate is available, but little is known about the factors that control the possible processes involved. The sandy clay loam soil with pH 10 and electrolytic conductivity 56 dS m^?1 was spiked with 1000 mg glucose-C kg^?1 soil (GLUCOSE pre-treatment), 200 mg NO₃^?-N kg^?1 soil (NITRATE pre-treatment), or left unamended (CONTROL pre-treatment) and conditioned for eight days. Pre-treated soil was then added with 1000 mg glucose-C kg^?1 soil and 200 mg NO₃^?-N kg^?1 soil and amended with ammonium (NH₄⁺) (AMM treatment) and l-glutamine (GLUT treatment), acetylene (C₂H₂) (ACE treatment), oxygen (O₂) (OXI treatment), left untreated (CON treatment) or sterilized. No abiotic factors affected concentrations of NH₄⁺, NO₂^? or NO₃^?. In the CONTROL pre-treatment, concentration of NO₃^? decreased 170 mg N kg^?1 soil within 72 h, in the GLUCOSE pre-treatment with 182 mg N kg^?1 soil within 2 h and in the NITRATE pre-treatment with 272 mg N kg^?1 soil within 168 h. Mean concentration of NO₂^? was 3.2 mg N kg^?1 soil in unamended soil, 5.7 mg N kg^?1 soil in the CONTROL pre-treatment, but >20 mg kg^?1 soil in the GLUCOSE pre-treatment and ≥40 mg kg^?1 in the NITRATE pre-treatment. The application of NO₃^? and glucose increased the mean concentration of NH₄⁺ compared to the unamended soil independently of pre-treatment. It was found that microorganisms in the alkaline saline soil of the former Lake Texcoco can reduce concentrations of NO₃^? while releasing NO₂^? under aerobic conditions when an easy decomposable substrate is available without it being directly related to microbial activity and this being more outspoken when glucose or nitrate were previously added.     

Lumbricus terrestris L. does not impact on the remediation efficiency of compost and biochar amendments

The aim of this study was to test the impact of compost and biochar, with or without earthworms, on the mobility and availability of metals, and on the growth of grass to re-vegetate contaminated soil from the Parys Mountain mining site, Anglesey. We also determined if the addition of earthworms compromises remediation efforts.In a laboratory experiment, contaminated soil (1343 mg Cu kg^?1, 2511 mg Pb kg^?1 and 262 mg Zn kg^?1) was remediated with compost and/or biochar. After 77 days Lumbricus terrestris L. earthworms were added to the treatment remediated with both compost and biochar, and left for 28 days. L. terrestris was not able to survive in the biochar, compost or unamended treatments. A germination and growth bioassay, using Agrostis capillaris (Common Bent) was then run on all treatments for 28 days.The combination of biochar and compost decreased water soluble Cu (from 5.6 to 0.2 mg kg^?1), Pb (from 0.17 to less than 0.007 mg kg^?1) and Zn (from 3.3 to 0.05 mg kg^?1) in the contaminated soil and increased the pH from 2.7 to 6.6. The addition of L. terrestris to this treatment had no effect on the concentration of the water soluble metals in the remediated soil.The compost was the only treatment that resulted in germination and growth of A. capillaris suitable for re-vegetation purposes. However, the combination of compost and biochar (with or without L. terrestris) produced the lowest concentrations of Cu (8 mg kg^?1) and Zn (36 mg kg^?1) in the aboveground biomass, lower than the compost treatment (15 mg Cu kg^?1 and 126 mg Zn kg^?1).The addition of biochar and compost both separately and as co-amendments was effective in reducing the mobility and availability of metals. The addition of L. terrestris did not re-mobilise previously sequestered metals.     

Effects of azoxystrobin,chlorothalonil, and ethoprophos on the reproduction of three terrestrial invertebrates using a natural Mediterranean soil

The potential terrestrial toxicity of three pesticides, azoxystrobin, chlorothalonil, and ethoprophos was evaluated using reproduction ecotoxicological tests with different non-target species: the collembolan Folsomia candida, the earthworm Eisenia andrei, and the enchytraeid Enchytraeus crypticus. All reproduction tests were performed with natural soil from a Mediterranean agricultural area (with no pesticide residues) in order to improve the relevance of laboratory data to field conditions. Controls were performed with natural and standard artificial soil (OECD 10% OM). The fungicide azoxystrobin showed the highest toxicity to earthworms (EC₅₀ = 42.0 mg a.i. kg⁻¹ dw soil). Collembolans were the most sensitive taxa in terms of sublethal effects of chlorothalonil with an EC₅₀ of 31.1 mg a.i. kg⁻¹ dw soil followed by the earthworms with an EC₅₀ of 40.9 mg a.i. kg⁻¹ dw soil. The insecticide ethoprophos was the most toxic to collembolans affecting their reproduction with an EC₅₀ of 0.027 mg a.i. kg⁻¹ dw soil. Enchytraeids were generally the least sensitive of the three species tested for long-term effects. Earthworms were not always the most sensitive species, emphasizing the need to increase the number of mandatory assays with key non-target organisms in the environmental risk assessment of pesticides.     

Accumulation of As,Pb, Zn,Cd and Cu and arbuscular mycorrhizal status in populations of Cynodon dactylon grown on metal-contaminated soils

Metal(loid) accumulation and arbuscular mycorrhizal (AM) status of the dominant plant species, Cynodon dactylon, growing at four multi-metal(loid)s-contaminated sites and an uncontaminated site of China were investigated. Up to 94.7 As mg kg^?1, 417 Pb mg kg^?1, 498 Zn mg kg^?1, 5.8 Cd mg kg^?1 and 27.7 Cu mg kg^?1 in shoots of C. dactylon were recorded. The plant was colonized consistently by AM fungi (33.0–65.5%) at both uncontaminated site and metal-contaminated sites. Based on morphological characteristics, fourteen species of AM fungi were identified in the rhizosphere of C. dactylon, with one belonging to the genus of Acaulospora and the other thirteen belonging to the genus of Glomus. Glomus etunicatum was the most common species associated with C. dactylon growing at metal-contaminated sites. Spore abundance in the rhizosphere of C. dactylon growing at the metal-contaminated soils (22–82 spores per 25 g soil) was significantly lower than that of the uncontaminated soils (371 spores per 25 g soil). However, AM fungal species diversity in the metal-contaminated soils was significantly higher than that in the uncontaminated soils. This is the first report of AM status in the rhizosphere of C. dactylon, the dominant plant survival in metal-contaminated soils. The investigation also suggests that phytorestoration of metal-contaminated sites might be facilitated using the appropriate plant with the aid of tolerant AM fungi.     

Removal of benzo (a) pyrene from soil using an endogeic earthworm Pontoscolex corethrurus ()

The endogeic earthworm Pontoscolex corethrurus (Müller, 1857) was the most abundant species (75%) in soil contaminated with hydrocarbons, mostly benzo(a)pyrene (BaP), in the state of Tabasco (Mexico). The earthworm P. corethrurus was tested for its capacity to remove 100 mg BaP kg⁻¹ from an Anthrosol soil (sterilized or not) and amended with legume Mucuna pruriens (L.) DC. var. utilis (Wall. ex Wight) Baker ex Burck (3%) or the grass Brachiaria humidicola (L.) DC (3%) (recently renamed as Urochloa humidicola (Rendle) Morrone & Zuloaga) in an aerobic incubation experiment. P. corethrurus removed 26.6 mg BaP kg⁻¹ from the sterilized soil and application of B. humidicola as feed increased this to 35.7 mg BaP kg⁻¹ and M. pruriens to 34.2 mg BaP kg⁻¹ after 112 days. The autochthonous microorganisms removed 9.1 mg BaP kg⁻¹ from the unsterilized soil and application of B. humidicola increased this to 18.0 mg BaP kg⁻¹ and M. pruriens to 11.2 mg BaP kg⁻¹. Adding P. corethrurus to the unsterilized soil accelerated the removal of BaP and 36.1 mg kg⁻¹ was dissipated from soil. It was found that the autochthonous microorganisms removed BaP from soil, but addition of P. corethrurus increased the dissipation 4-fold. The endogeic earthworm P. corethrurus can thus be used to remediate hydrocarbon-contaminated soils in tropical regions.     

Impact of high cadmium and nickel soil concentration on selected physiological parameters of Arundo donax L.

The purpose of this study was to investigate the effects of high cadmium and nickel soil concentrations on selected physiological parameters of Arundo donax L. A 2-year pot experiment was held in the field and the pots were irrigated with aqueous solutions of Cd and Ni in concentrations of 5, 50 and 100 ppm, against the control (tap water). At the end of the cultivation periods the pots soil was divided into three equal zones and total and NH₄OAc extractable Cd and Ni concentrations were determined. The top zone exhibited the highest metal content. Cadmium and nickel total concentrations at the end of the experiment were up to 973.8 mg kg⁻¹ and 2543.3 mg kg⁻¹ respectively, while NH₄OAc extractable Cd was up to 291.7 mg kg⁻¹ and Ni up to 510.3 mg kg⁻¹. Stomatal conductance ranged between 0.3 and 0.8 mol CO₂ m⁻² s⁻¹, intercellular CO₂ concentration ranged between 212.9 and 243.0 ppm CO₂, stomatal resistance between 0.6 and 1.3 s cm⁻¹, chlorophyll content (SPAD values) between 46.3 and 57.0 and chlorophyll fluorescence (F_v/F_m) ranged between 0.8 and 0.9. All studied physiological parameters did not show statistically significant differences among control and heavy metal treated plants for both years; therefore, high soil cadmium and nickel concentration did not inhibit stomatal opening and did not affect the function of the photosynthetic machine of A. donax plants.     

Molecular and physiological bacterial diversity of a semi-arid soil contaminated with different levels of formulated atrazine

A semi-arid soil treated with different concentrations of formulated atrazine in a laboratory experiment was studied over 45 days, by different biological and molecular parameters (bacterial enumeration (cfu), community level physiological profiles (CLPPs) measured by Biolog^® and denaturing gradient gel electrophoresis (DGGE)), to study the bacterial community diversity.Formulated atrazine was almost totally degraded at different concentrations after this incubation time. The number of colony forming units (cfu) for soils with 100 and 1000 mg kg⁻¹ atrazine was significantly (p ≤ 0.05) higher than for the control, 1 and 10 mg kg⁻¹ treatments. DGGE banding patterns showed that regardless of time elapsed, concentrations of 10, 100 and 1000 mg kg⁻¹ atrazine in soil, affected the bacterial community compared to control and 1 mg kg⁻¹.The Shannon diversity index (H′) based on CLPP data showed a significant (p ≤ 0.05) decrease at atrazine concentrations of 100 and 1000 mg kg⁻¹. The Shannon diversity indices for different guilds of source carbon and the parameters K and r (based on the kinetics of colour formation rather than on the degree of colour development) were related to guilds of carbon substrates and atrazine concentration at a sampling time. The parameter K was very sensitive to atrazine effects on microbial communities.These biological and molecular parameters can be used to monitor changes in soils treated with atrazine at different concentrations, even when the pesticide is degraded.     

Earthworm colonisation of abandoned arable soil polluted by copper

The distribution, density and biomass of earthworms were investigated at the copper polluted site, Hygum (Denmark). In 1994, shortly after farming of the area was abandoned, only four earthworm species were present and their distribution was restricted to areas where copper concentration did not exceed 200 mg kg^?1 dry soil. Sixteen years later (in 2010), without any agricultural activity, ten species of earthworms were found, in particular, epigeic species were present where soil copper concentrations reached >1000 mg kg^?1 dry soil.     

Climatic influences on active fractions of soil organic matter

Biologically active fractions of soil organic matter are important in understanding decomposition potential of organic materials, nutrient cycling dynamics, and biophysical manipulation of soil structure. We evaluated the quantitative relationships among potential C and net N mineralization, soil microbial biomass C (SMBC), and soil organic C (SOC) under four contrasting climatic conditions. Mean SOC values were 28±11 mg g⁻¹ (n=24) in a frigid–dry region (Alberta/British Columbia), 25±5 mg g⁻¹ (n=12) in a frigid–wet region (Maine), 11±4 mg g⁻¹ (n=117) in a thermic–dry region (Texas), and 12±5 mg g⁻¹ (n=131) in a thermic–wet region (Georgia). Higher mean annual temperature resulted in consistently greater basal soil respiration (1.7 vs 0.8 mg CO₂–C g⁻¹ SOC d⁻¹ in the thermic compared with the frigid regions, P<0.001), greater net N mineralization (2.8 vs 1.3 mg inorganic N g⁻¹ SOC 24 d⁻¹, P<0.001), and greater SMBC (53 vs 21 mg SMBC g⁻¹ SOC, P<0.001). Specific respiratory activity of SMBC was, however, consistently lower in the thermic than in the frigid regions (29 vs 34 mg CO₂–C g⁻¹ SMBC d⁻¹, P<0.01). Higher mean annual precipitation resulted in consistently lower basal soil respiration (1.1 vs 1.3 mg CO₂–C g⁻¹ SOC d⁻¹ in the wet compared with the dry regions, P<0.01) and lower SMBC (31 vs 43 mg SMBC g⁻¹ SOC, P<0.001), but had inconsistent effects on net N mineralization that depended upon temperature regime. Specific respiratory activity of SMBC was consistently greater in the wet than the dry regions (≈33 vs 29 mg CO₂–C g⁻¹ SMBC d⁻¹, P<0.01). Although the thermic regions were not able to retain as high a level of SOC as the frigid regions, due likely to high annual decomposition rates, biologically active soil fractions were as high per mass of soil and even 2–3-times greater per unit of SOC in the thermic compared with the frigid regions. These results suggest that macroclimate has a large impact on the portion of soil organic matter that is potentially active, but a relatively small impact on the specific respiratory activity of SMBC.     

Fate of sulfadiazine administered to pigs and its quantitative effect on the dynamics of bacterial resistance genes in manure and manured soil

The veterinary antibiotic sulfadiazine (SDZ), labelled by ¹⁴C, was administered to pigs to follow the fate of the drug and its metabolites in manure and manure-amended soil, and to investigate the dynamics of drug effects on resistance genes and bacterial communities. In the manure sampled over 10 days, more than 96% of the drug was found as parent compound or metabolites N-acetyl-SDZ and 4-hydroxy-SDZ. While the manure was stored the concentration of SDZ increased by 42% due to deacetylation of the metabolite N-acetyl-SDZ, whereas the minor metabolite 4-hydroxy-SDZ kept constant. In the soil the extractable amounts of the compounds decreased exponentially to less than 1 mg kg^?1 within 11 days after manure amendment. The abundances of SDZ resistance genes sul1 and sul2 were determined by qPCR relative to 16S rRNA genes in total DNA from manure and manure-amended soil. In manure both genes increased exponentially in copy number during the first 60 days of storage, suggesting preferential growth of resistant populations. However, the abundance of sul1 and sul2 decreased below 10^?5 copies per 16S rRNA gene after 175 days. With manure high amounts of sul1 and sul2 were introduced into the soil which were reduced by more than 10 times within 24 days. Thereafter, sul1 was stably maintained in soil, while sul2 further decreased between day 60 and day 165. A mathematical model was developed that could well explain the time course of sul gene abundance by considering the cost of sul genes, horizontal gene transfer, and selection of the resistant populations in the presence of SDZ. Modelling revealed a selective effect of SDZ on sul2 even at low concentrations down to 0.15 mg kg^?1 soil. Bacterial community profiles of manure and manure-amended soil were distinct, indicating that bacteria introduced with manure do not become prominent in soil. The composition of the bacterial community in manure constantly changed during storage, but mainly during the first 10 days. Profiles of soil bacterial communities revealed only a transient perturbation by manure containing SDZ.     

Incongruous variation of denitrifying bacterial communities as soil N level rises in Canadian canola fields

Soil N fertilization stimulates the activity of the soil bacterial species specialized in performing the different steps of the denitrification processes. Different responses of these bacterial denitrifiers to soil N management could alter the efficiency of reduction of the greenhouse gas N₂O into N₂ gas in cultivated fields. We used next generation sequencing to show how raising the soil N fertility of Canadian canola fields differentially modifies the diversity and composition of nitrite reductase (nirK and nirS) and nitrous oxide reductase (nosZ) gene-carrying denitrifying bacterial communities, based on a randomized complete blocks field experiment. Raising soil N levels increased up to 60% the ratio of the nirK to nirS genes, the two nitrite reductase coding genes, in the Brown soil and up to 300% in the Black soil, but this ratio was unaffected in the Dark Brown soil. Raising soil N levels also increased the diversity of the bacteria carrying the nitrite reductase gene nirK (Simpson index, P = 0.0417 and Shannon index, 0.0181), and changed the proportions of the six dominant phyla hosting nirK, nirS, and nosZ gene-carrying bacteria. The level of soil copper (Cu) and the abundance of nirK gene, which codes for a Cu-dependent nitrite reductase, were positively related in the Brown (P = 0.0060, R² = 0.48) and Dark Brown (0.0199, R² = 0.59) soils, but not in the Black soil. The level of total diversity of the denitrifying communities tended to remain constant as N fertilization induced shifts in the composition of these denitrifying communities. Together, our results indicate that higher N fertilizer rate increases the potential risk of nitrous oxide (N₂O) emission from canola fields by promoting the proliferation of the mostly adaptive N₂O-producing over the less adaptive N₂O-reducing bacterial community.     

Microbial utilization and mineralization of [14C]glucose added in six orders of concentration to soil

The substrate availability for microbial biomass (MB) in soil is crucial for microbial biomass activity. Due to the fast microbial decomposition and the permanent production of easily available substrates in the rooted top soil mainly by plants during photosynthesis, easily available substrates make a very important contribution to many soil processes including soil organic matter turnover, microbial growth and maintenance, aggregate stabilization, CO₂ efflux, etc. Naturally occurring concentrations of easily available substances are low, ranging from 0.1 μM in soils free of roots and plant residues to 80 mM in root cells. We investigated the effect of adding ¹⁴C-labelled glucose at concentrations spanning the 6 orders of magnitude naturally occurring concentrations on glucose uptake and mineralization by microbial biomass. A positive correlation between the amount of added glucose and its portion mineralized to CO₂ was observed: After 22 days, from 26% to 44% of the added 0.0009 to 257 μg glucose C g^?1 soil was mineralized. The dependence of glucose mineralization on its amount can be described with two functions. Up to 2.6 μg glucose C g^?1 soil (corresponds to 0.78% of initial microbial biomass C), glucose mineralization increased with the slope of 1.8% more mineralized glucose C per 1 μg C added, accompanied by an increasing incorporation of glucose C into MB. An increased spatial contact between micro-organisms and glucose molecules with increasing concentration may be responsible for this fast increase in mineralization rates (at glucose additions <2.6 μg C g^?1). At glucose additions higher than 2.6 μg C g^?1 soil, however, the increase of the glucose mineralization per 1 μg added glucose was much smaller as at additions below 2.6 μg C g^?1 soil and was accompanied by decreasing portions of glucose ¹⁴C incorporated into microbial biomass. This supports the hypothesis of decreasing efficiency of glucose utilization by MB in response to increased substrate availability in the range 2.6–257 μg C g^?1 (=0.78–78% of microbial biomass C). At low glucose amounts, it was mainly stored in a chloroform-labile microbial pool, but not readily mineralized to CO₂. The addition of 257 μg glucose C g^?1 soil (0.78 μg C glucose μg^?1 C micro-organisms) caused a lag phase in mineralization of 19 h, indicating that glucose mineralization was not limited by the substrate availability but by the amount of MB which is typical for 2nd order kinetics.     

Emission of nitrous oxide from hydrocarbon contaminated soil amended with waste water sludge and earthworms

Soils in Mexico are often contaminated with hydrocarbons and addition of waste water sludge and earthworms accelerates their removal. However, little is known how contamination and subsequent bioremediation affects emissions of N₂O and CO₂. A laboratory study was done to investigate the effect of waste water sludge and the earthworm Eisenia fetida on emission of N₂O and CO₂ in a sandy loam soil contaminated with the polycyclic aromatic hydrocarbons (PAHs): phenanthrene, anthracene and benzo(a)pyrene. Emissions of N₂O and CO₂, and concentrations of inorganic N (ammonium (NH₄⁺), nitrite (NO₂^?) nitrate (NO₃^?)) were monitored after 0, 5, 24, 72 and 168 h. Adding E. fetida to the PAHs contaminated soil increased CO₂ production rate significantly 2.0 times independent of the addition of sludge. The N₂O emission rate from unamended soil expressed on a daily base was 5 μg N kg^?1 d^?1 for the first 2 h and increased to a maximum of 325 μg N kg^?1 d^?1 after 48 h and then decreased to 10 μg N kg^?1 d^?1 after 168 h. Addition of PAHs, E. fetida or PAHs + E. fetida had no significant effect on the N₂O emission rate. Adding sludge to the soil sharply increased the N₂O emission rate to >400 μg N kg^?1 d^?1 for the entire incubation with a maximum of 1134 μg N kg^?1 d^?1 after 48 h. Addition of E. fetida, PAHs or PAHs + E. fetida to the sludge-amended soil reduced the N₂O emission rate significantly compared to soil amended with sludge after 24 h. It was found that contaminating soil with PAHs and adding earthworms had no effect on emissions of N₂O. Emission of N₂O, however, increased in sludge-amended soil, but addition of earthworms to this soil and contamination reduced it.     

Effects of nonylphenols on soil microbial activity and water retention

The main aim of this study is to analyze the influence of 4-nonylphenol (NP) on soil water retention and biological activity. Two doses of 4-nonylphenol (25 and 50 mg kg⁻¹) were tested in a loam soil with and without peat amendment. In general, one week after the start of the experiment, the soil water content retained at −0.75 MPa of soil suction was 18% higher in the soil amended and its basal respiration (BR) was 15% higher than soil without peat. In contrast, the microbial activity indices (CM: coefficient of mineralization or BR:total organic carbon (TOC) ratio; Cmic:Corg: microbial biomass carbon (MBC):TOC ratio; qCO₂: metabolic quotient or BR:MBC ratio) were higher in the soil without peat, compared to the soil amended with peat. On the other hand, the addition of NP to soil was able to modify soil biological but not physical (water retention, desorption) properties. When soil was amended with peat, MBC was reduced one week after applying NP. In contrast, no effects of NP on MBC were observed in the soil without peat. BR was reduced by 16% one week after applying 50 mg kg⁻¹ of NP to soil with peat, and was increased by 46% one week after applying 25 mg kg⁻¹ of NP to soil without peat. The effects of NP on MBC and BR could be associated more with the adsorption of NP by soil organic matter, while changes in CM or Cmic:Corg ratio were more closely related to changes in soil water retention. The potential toxic effects of NP (high qCO₂ values) were only observed in the absence of peat amendments. Peat addition reduced NP toxic effects on microorganisms.     

Effects of cultivation on soil microbiological properties in a freshwater marsh soil in Northeast China

The Sanjiang Plain has become an intensive area of land use/cover change in China. However, little is known about the effect of cultivation on soil microbiological properties in this freshwater marsh ecosystem. Our objective was to evaluate the effect of cultivation on mineralizable, microbial biomass, and total C in the Sanjiang Plain of Northeast China. Soil microbial biomass C (MBC) was 4346 ± 309 mg kg⁻¹ in undisturbed marsh and 229 mg kg⁻¹ in soil cultivated for 15 years. Undisturbed marsh soil had the highest microbial quotient (3.64%), which declined with increasing cultivation time (R² = 0.97, p < 0.01). Metabolic quotient increased with increasing cultivation time. Soil C mineralization in undisturbed marsh was 3.5 times that in soil cultivated for 1 year, and was 12 times that in soil cultivated for 15 years. Cultivation strongly affected measured soil microbiological properties.     

Mercury affects the distribution of culturable species of Pseudomonas in soil

Pseudomonas bacteria isolated during 52 days on Gould's S1 agar from soil spiked with 0, 3.5 and 15 mg Hg(II) kg soil⁻¹ were characterised to reveal whether mercury affected them differently. Isolates from the treatments with 0 and 15 mg Hg kg⁻¹ were characterised using FT-IR characterisation and subsequent 16S rDNA partial sequencing of representative isolates. To verify the selectivity of Gould's S1 agar and the FT-IR characterisation, all 450 isolates were subjected to the following tests: Gram-determination, catalase and oxidase activity, pigment production on PDA and growth at different temperatures. Furthermore, the isolates were tested for their ability to grow on agar amended with 10 mg Hg kg⁻¹ as an indication of mercury resistance. We found that up to 80% of the isolates in soil amended with 15 mg Hg kg⁻¹ were mercury-resistant, whereas only up to 20% were resistant in the treatments with 0 and 3.5 mg Hg kg⁻¹. We found two groups of Pseudomonas, which probably represent non-described species since they did not group closely with any known species of Pseudomonas in the dendrogram. Hg-enhanced isolates were closely related to P. frederiksbergensis. Furthermore, Hg resistance was almost exclusively restricted to P. frederiksbergensis and P. migulae groups. We conclude that Hg caused a shift in the dominating species of culturable Pseudomonas.     

Effects of dazomet on soil organisms and recolonisation of fumigated soil

Fumigation is a common practice to control soil pathogens, but little is known about the impacts of fumigation on other soil biota groups. The purpose of this study was to investigate the effects of fumigation on soil biota, including microorganisms, nematodes, and microarthropods. Bacteria were the most resistant group and some survived following treatment with 2000 mg kg⁻¹ dazomet. Some soil fungi survived 100 mg kg⁻¹ dazomet, although they were mainly Trichoderma. The fungi pathogenic to ginseng were all killed at 100 mg kg⁻¹, and showed both inter- and intra-species variation with respect to dazomet susceptibility. Among the nematodes, Aphelenchus was relatively resistant. The results suggested that susceptibility of soil organisms to dazomet differs between species, and that tolerant organisms may engage in recolonisation. In microcosm experiments, the microbial biomass and community were assessed using phospholipid fatty acid (PLFA) analysis while recolonisation of soil organisms was controlled by mesh size. The bacterial PLFA levels were changed little after fumigation, whereas the fungal PLFA levels gradually increased after fumigation. Principal analysis of the PLFA levels and the ratio of gram-negative to gram-positive bacteria showed that fumigation altered the microbial community. The number of nematodes did not recover even at 12 weeks after fumigation. The increased Collembolan numbers suggest that fumigated soil could be recolonised by specific organisms that have adapted to the conditions. In field experiments, we tested the ability of organic materials to enhance the recolonisation of fumigated soil by soil organisms. Bean powder and rice bran increased the microbial PLFA levels and nematode numbers at 6 weeks and 12 weeks after treatment, and the abundance of nematodes continued to increase 42 weeks after fumigation. The abundance of microarthropods was only slightly affected by the presence of the organic materials. We suggest that treating fumigated soils with organic materials is an effective technique to promote soil organism numbers. In addition, Trichoderma was observed to be relatively resistant to fumigation, and therefore, we propose that the fumigation effect can be improved by using a combination of resistant Trichoderma and dazomet.     

Population size of indigenous Rhizobium leguminosarum biovar trifolii in long-term field experiments with sewage sludge cake,metal-amended liquid sludge or metal salts: Effects of zinc,copper and cadmium

There is conflicting evidence, and therefore continuing concern, as to whether metals in sewage sludge are deleterious to soil microbial processes and long-term agricultural productivity. Nine field experiments with sewage sludge cakes, three with metal-amended liquid sludges and three with inorganic metal salts were set up across Britain in 1994 to give individual metal dose–response treatments to try to answer this question. This study reports on the effects of Zn, Cu and Cd on the population size of Rhizobium leguminosarum biovar trifolii, a nitrogen fixing symbiont of white clover (Trifolium repens), in soils from these experiments over 11 years. Significant (P < 0.05) reductions in indigenous rhizobial numbers occurred on the Zn metal dose–response treatments at eight of the sludge cake sites in 2005, but few consistent effects were evident on the Cu or Cd metal dose–response treatments during the 11-year monitoring period. The soil total Zn concentrations where effects occurred were near to the UK statutory limit of 300 mg kg^?1 for soils receiving sewage sludge. No significant reductions occurred in any treatments on the metal-amended liquid sludge or inorganic metal salt experiments in which the metals would be expected to be in a more bioavailable form, even after 11 years. The effects in the sludge cake experiments were related consistently with soil total Zn, with no recovery to date. The reductions in clover rhizobial numbers in the sludge cake experiments were due to Zn effects on free-living rhizobia in the soil, with gradual die-off over a long time with increasing soil total Zn concentrations. Currently, no consistent adverse effects on rhizobia have been seen at the UK limits for Cu and Cd of 135 and 3 mg kg^?1, respectively.     

Changes in soil microbial biomass and functional diversity with a nitrogen gradient in soil columns

Fertilization generates nutrient patches that may impact soil microbial activity. In this study, nitrogen patches were generated by adding ammonium sulfate or urea to soil columns (length 25 cm; internal diameter 7.2 cm). Changes in nitrogen transformation, soil microbial biomass, and microbial functional diversity with the nitrogen gradients were investigated to evaluate the response of microbial activity to chemical fertilizer nutrient patches. After applying of ammonium sulfate or urea, the added nitrogen migrated about 7 cm. Microbial biomass carbon (MBC) was lower in fertilized soil than in the control (CK) treatment at the same soil layers. MBC increased with soil depth while microbial biomass nitrogen (MBN) decreased. BIOLOG analysis indicated that the average well color development (AWCD) and functional diversity indices of the microbial communities were lower in the 1 cm and 2 cm soil layers after application of ammonium sulfate; the highest values were in the 3 cm soil layer. AWCD and Shannon indices from the 1 to 5 cm soil layers were higher than those from other soil layers under urea application. Both principal component analysis and carbon substrate utilization analysis showed significant separation of soil microbial communities among different soil layers under application of ammonium sulfate or urea. Microbial activity was substantially decreased when NH₄⁺-N concentration was higher than 528.5 mg kg⁻¹ (1–3 cm soil layer under ammonium sulfate application) or 536.8 mg kg⁻¹ (1 cm soil layer under urea application). These findings indicated that changes in soil microbial biomass and microbial functional diversity can occur with a nitrogen gradient. The extent of changes depends on the nitrogen concentration and the form of inorganic fertilizer.