Estimation of microbial biomass and activity in soil using microcalorimetry

Relationships between the rate of heat output from soil, the rate of respiration and the soil microbial biomass were investigated for 25 soils from northern Britain. The rate of heat output, measured in a Calvet microcalorimeter at 22°C, correlated well with the rate of carbon dioxide respiration. The average amount of heat evolved per cm³ of gas respired. 21.1 J cm^?3, suggests that the biomass metabolism was largely aerobic. The rate of heat output per unit of total microbial biomass was remarkably uniform over a wide range of soils, but showed differences depending upon whether the soil had been stored or amended. Mineral soils that had been stored at 4°C had the lowest heat output, 12.0 mW g^?1 biomass C, compared with a mean of 20.4 mW g^?1 biomass C for freshly-collected soils. Amendment with glucose (0.5% w/w) caused an immediate increase in respiration and heat output, up to 59.4 mW g^?1 biomass C for stored soils and 188.2 mW g^?1 biomass C for freshly collected soils. There was a consistent relationship between the biomass and the rate of heat output from freshly collected and amended mineral and organic soils which gave a linear fit using log transformed data: y= 0.6970+ 1.025x (r= 0.98, P < 0.001) (y=log₁₀ biomass C, μgC g^?1; x=log₁₀ rate of heat output at 22°C, μW g^?1). The overall relationship between biomass and the rate of heat output for all the amended samples was: 1 g biomass C= 180.05 ± 34.61 mW.     

农艺措施对降低污染土壤重金属活性的影响

本文利用小冶炼厂重金属污染的农田为对象,研究了不同农艺措施对土壤重金属活性及植物累积的差异,并对不同农艺措施改良效果作了评价,为植物修复重金属污染土壤选择相应的农艺措施提供科学依据。     

Soil microbial activity measured by microcalorimetry in response to long-term fertilization regimes and available phosphorous on heat evolution

Microcalorimetric technique was used to investigate the effects of balanced versus nutrient-deficiency fertilization on soil microbial activity in a long-term (19-year) fertilizer experiment. The number of microorganisms in soils was measured by viable cell count, and the power-time curves were recorded for soil samples supplemented with glucose and ammonium sulphate, also with or without sodium dihydrogen phosphate for P-deficiency fertilization. Both the bacterial and the fungal populations were significantly higher (P < 0.05) under balanced fertilization than under nutrient-deficiency fertilization. The microbial activity presented by heat dissipation per cell unit indicated that microorganisms under balanced fertilizer treatments had more efficient metabolism, while decreased microbial activity under nutrient-deficiency treatments was firstly due to soil available P, followed by N and K (P < 0.05). In addition, microbial growth in soils under P-deficiency fertilization was stimulated by adding available P, while the lower growth rate, less peak heat, and longer peak time all indicated the low activity of soil microorganisms. We emphasize the importance of balanced fertilization, as well as the role of available P, in maintaining and promoting soil microbial activity.     

Effects of agricultural biostimulants on soil microbial activity and nitrogen dynamics

We investigated the effects of two commercially available soil biostimulants, designated Z93 and W91, on key microbial and nutrient cycling processes in the soil, by conducting short-term (1 week) and longer-term (8 weeks) soil incubations in the laboratory. In the short-term soil incubations, the two compounds differed in their effects on microbial activity: Z93 was effective over a wide range, stimulating substrate-induced respiration (SIR) and dehydrogenase activity (DHA) at remarkably low concentrations (0.5–500 nl/g soil); W91 stimulated SIR at these concentrations, but also inhibited DHA. In longer-term soil incubations, we amended batches of soil with either finely-ground alfalfa leaves, wheat straw, or added no amendments, to alter patterns of soil nitrogen mineralization and immobilization. We treated these soils with Z93 and W91 at two concentrations (0.005 and 0.5 μl/g soil), and incubated them for up to 8 weeks. These extremely low doses of both Z93 and W91 influenced soil SIR, DHA, and cellulase activity significantly (P<0.05). Both compounds also influenced soil nitrogen dynamics significantly; the extent depending upon the quality of the organic amendments. In the alfalfa-amended soil there was a steep increase in NO₃-N concentration during the incubation due to the rapid mineralization of nitrogen-rich alfalfa material. However, in this soil, both Z93 and W91 reduced NO₃-N concentrations greatly after 56 days. In the straw-amended soil, mineral nitrogen concentrations were very low, probably due to rapid immobilization of nitrogen by microbial biomass. In this soil, treatment with both compounds decreased microbial biomass nitrogen and increased dissolved organic nitrogen (DON), relative to that in the controls. Our results suggest that the two biostimulants can stimulate both the breakdown and mineralization of soil organic materials, perhaps by selectively inhibiting or stimulating particular components of the microbial community, leading to lasting (8 weeks or longer) increases in soil nitrogen availability.     

Effect of long-term fertilizers and manure application on microbial biomass and microbial activity of a tropical agricultural soil

We investigated some aspects of soil quality and community-level physiological profiles (CLPP) of bacteria in soil under a long-term (37 years) trial with either exclusive inorganic fertilizers or fertilizers combined with farmyard manure cultivated with jute–rice–wheat system. The treatments consisted of 100% recommended dose (RD) of NPK, 150% RD of NPK, 100% RD of N, 100% RD of NPK + FYM (10 t ha⁻¹ year⁻¹), and untreated control. Long-term application of 150% RD of NPK lowered the soil pH considerably while the soils in the other treatments remained near neutral. The 100% RD of NPK + FYM treated plot showed significantly highest accumulation of organic carbon, total nitrogen, microbial biomass carbon, basal soil respiration, and fluorescein diacetate hydrolyzing activity among the treatments. CLPP analysis in Biolog Ecoplates revealed that utilization of carbohydrates was enhanced in all input treated regimes, while the same for polymers, carboxylic acids, amino acids, and amines/amides were similar or less than the untreated control. However, within these groups of carbon sources, heterogeneity of individual substrate utilization between treatments was also noted. Taken together, addition of organic supplements showed significantly increased microbial biomass carbon and microbial activity, but input of nutrient supplements, both inorganic and organic, only marginally affected the overall substrate utilization pattern of soil microorganisms.     

Impact of soil fumigation practices on soil nematodes and microbial biomass

This study was designed to understand the impact of methyl bromide (MB) (CHaBr) and its alternatives on both free-living and root-knot nematodes in the soil. A randomized complete block experiment with six treatments and 4 replicates (each replicate in a separate greenhouse) was established in Qingzhou, Shandong Province, China. In addition to MB and untreated control (CK) treatments there were four alternative soil fumigation practices including MB virtually impermeable films (VIF), metam sodium (MS), MS VIF and soil solarization combined with selected biological control agents (SS BCA). Two tomato (Lycopersicum esculentum Mill.) cultivars, cv. Maofen-802 from the Xian Institute of Vegetable Science, China, and cv. AF179 Brillante from the Israeli Hazera Quality Seeds, were selected as test crops. The results indicated that Rhabditidae was the most dominant population with percentage abundance as high as 85% of the total number of identified free-living nematodes, followed by that of Cephalobidae. Methyl bromide and its alternatives except for the non-chemical SS BCA treatment controlled the target pest, root-knot nematodes. Also, the impact of the three chemical alternatives on free-living nematode number and functional group abundance was similar to the impact associated with a typical methyl bromide application. Chemical fumigation practices, especially that with MB, significantly reduced the number of nematodes in the soil and simultaneously significantly reduced the number of nematode genera thereby reducing nematode diversity. All the four soil chemical fumigation activities decreased soil microbial biomass and had an obvious initial impact on microorganism biomass. Furthermore, both plant-parasitic and fungivore nematodes were positively correlated with soil microbial biomass.     

Soil microbial diversity and activity in a Mediterranean olive orchard using sustainable agricultural practices

Sustainable soil management of orchards can have positive effects on both soils and crop yields due to increases in microbial biomass, activity and complexity. The aim of this study was to investigate medium‐term effects (12 yr) of two different management practices termed ‘sustainable’ (ST) and ‘conventional’ (CT) on soil microbial composition and metabolic diversity of a rainfed mature olive orchard in Southern Italy. ST included no‐till, self‐seeding weeds (mainly graminaceous and leguminosae), and mulch derived from olive tree prunings, whilst CT was managed by frequent tillage and included severe pruning with residues removed from the orchard. Microbial analyses were carried out by culture‐dependent methods (microbial cultures and Biolog^®). Molecular methods were used to confirm the identification by light microscopy of the isolates of fungi and Streptomyces. Significantly more culturable fungi and bacteria were found in ST than in CT. The number of fungal groups in ST was also significantly greater than in CT. Overall and substrate‐specific Biolog^® metabolic diversity indices of microbial communities and soil enzyme activities were greater in ST. The results demonstrate that soil micro‐organisms respond positively to sustainable orchard management characterized by periodic applications of locally derived organic matter. This study confirms the need to encourage farmers with orchards in the Mediterranean basin to practise soil management based on organic matter inputs associated with zero tillage to improve soil functionality.     

Impact of agricultural land use on distribution of microbial biomass and activity within soil aggregates

The presence of aggregates of various sizes in the soil is an important condition for soil carbon sequestration. In this system, microbial biomass is a key link. This work was devoted to the study of the influence of land use systems on the distribution of SOС, MB-SIR, microbial activity and eco-physiological indices (qCO₂, QR, MB-SIR/SOС and qCO₂/SOС) in relation to the size of soil aggregates. The distribution of SOС, MB-SIR and mineralization activity among the aggregates was heterogeneous. In the soil of crop rotation, high mineralization activity and MB-SIR were found in the aggregates 0.5–0.1 mm, in the monoculture soil in aggregates <0.1 mm and in the control soil in the aggregates 1–0.25 mm. There was a general trend towards a decrease in microbial activity, MB-SIR and SOС availability with an increase in aggregate size. In agricultural soils, microbial activity was determined by large aggregates (>5 mm), while in the control soil, by the aggregates 5–1 mm. Depending on the type of site and the size of aggregates, the differences in microbial metabolism were revealed. The qCO₂ and QR values decreased, and the MB-SIR/SOС and qCO₂/SOС increased in the series: control soil > crop rotation > monoculture. In the control soil, the values of the eco-physiological indices decreased with decreasing aggregate size. And vice versa, in agricultural soils, these parameters were the highest in the microaggregates (<0.25 mm). The monoculture soil, in contrast to the control soil and crop rotation soil, turned out to be more energy efficient.     

Wetting of agricultural soil measured by a simplified capillary rise technique

We describe the use of a capillary rise method to measure the wettability of 10 samples of agricultural soil from Rothamsted long-term experimental sites. The samples have very similar clay contents, but organic carbon (C) contents range from 11.5 to 31.2 g kg⁻¹. Their wetting rates were interpreted by an improved method of data analysis, consistent with the Washburn equation, and showed an increase in the effective contact angle between the water meniscus and the soil with increasing C content. This corresponds to a decrease in wettability with increasing C content, and accords with other results reported in the literature. By contrast with water, we found that capillary rise for n -hexane into soil did not depend on the soil's bulk density or C content. A priori calculations of the expected wetting rates from fluid properties and an effective hydraulic radius estimated by other methods gave magnitudes and trends that agreed with our experimental data. The results show that estimates of effective hydraulic radius can provide a useful approximation for characterizing soil wetting, but that further modelling should be carried out.     

Influence of agricultural traffic and crop management on collembola and microbial biomass in arable soil

Collembola and microbial biomass C were investigated in a field experiment with controlled agricultural traffic and crop rotation over a period of 27 months. The wheel-induced compactive efforts were applied according to management practices within the crop rotation of sugar beet, winter wheat, and winter barley. Increasing wheel traffic produced increasing soil compaction, mainly due to a reduction in surface soil porosity. Increasing soil compaction was accompanied by a decrease in microbial biomass C and the density of collembola. The influence of soil compaction on microbial biomass C was smaller than that of the standing crop. However, for collembola, especially euedaphic species, a reduction in pore space appeared to be of more importance than the effects of a standing crop. Within the crop rotation, microbial biomass C and the density of collembola increased in the order sugar beet, winter wheat, and winter barley.     

Influence of microbial activity and soil moisture on herbicide immobilization in soils

Mobility, extractability, and disappearance of the herbicides diuron, terbuthylazine, metolachlor, and pendimethalin were examined in incubation experiments with two topsoil samples of different natural microbial activity and after sterilization. Soil moisture was held constant at 10, 40, and 60 % WHC. In other variants, the soil water content was changed during the incubation. The four herbicides reveal a fairly different extent of microbial and chemical degradation and immobilization. The herbicide mobility – expressed by coefficients of partition between adsorbed and dissolved herbicide amounts – decreases at a lower rate and extent, when the microbial activity is low or the soil is sterile. With increasing initial soil moisture, also herbicide mobility and extractability increase; but in the course of time, abiotic immobilization occurs to a higher extent. When soil moisture changes during the incubation, formerly non‐extractable herbicide fractions (up to 40 % of the applied amounts) become extractable. Kinetics of herbicide immobilization follow an empirical sigmoidal function, which describes three periods of immobilization. The three‐period shape of the curve and its possible reasons are discussed for the data of the incubation experiments as well as for the results of a long‐term field trial with diuron.     

Impact of agricultural practices on the size and activity of the microbial biomass in a long-term field experiment

The Dehérain long-term field experiment was initiated in 1875 to study the impact of fertilization on a wheat-sugarbeet rotation. In 1987, the rotation was stopped to be replaced by continuous maize. Crop residues were soil-incorporated and the mineral fertilization was doubled in some plots. The impact of those changes on the microbial biomass and activity are presented. In spring 1987, the soil was still in a steady-state condition corresponding to the rotation. The microbial biomass was correlated with total organic C and decreased in the order farmyard manure>mineral NPK>unfertilized control. Microbial specific respiratory activity was higher in the unfertilized treatments. The soil biomass was closely related to soil N plant uptake. In 1989, after 2 years of maize and crop residue incorporation, the steady-state condition corresponding to the previous agricultural practices disappeared. So did the relationship between the biomass and total organic C, and the soil N plant uptake. Biomass specific respiratory activity increased because of low efficiency in the use of maize residues by microbes under N stress.     

生物有机肥对土壤微生物活性的影响

通过两次连续温室玉米盆栽试验,研究了施用具有调节微生物功能的生物有机肥对土壤微生物数量与活性的影响,并利用传统平板计数法与BIOLOGECO方法相结合研究生物有机肥对土壤微生物生态的影响。结果表明,与化肥相比,施用生物有机肥可显著提高土壤微生物中3大菌群的数量;AWCD值及微生物对不同碳底物利用水平的测定结果表明,施用生物有机肥可明显提高土壤微生物对碳源的利用率,尤其土壤中的羧酸、胺类和其他类碳源等。表明生物有机肥的施用能增加土壤微生物利用碳源能力,改善微生物营养条件,使微生物保持较高活性,提高土壤微生物多样性。     

Influence of polychlorinated biphenyls on microbial biomass and its activity in grassland soil

Microbial biomass content, soil respiration and biomass specific respiration rate were measured in two parts of an area polluted by a municipal waste incinerator [polychlorinated biphenyls (PCBs) from combustion processes]. The soils in the studied parts differed significantly only in their levels of PCBs. The concentration of PCBs found in a control plot (4.4 ng g^-1 soil) can be regarded as a background value while the polluted plot contained an increased amount of PCBs (14.0 ng g^-1 soil). A significantly lower microbial biomass (decreased by 23%, based on the chloroform-fumigation extraction technique) and a lower specific respiration rate (decreased by 14%) were observed in the polluted plot in comparison with the control plot at the end of experimental period (1992–1994). Furthermore, a lower ability of microorganisms in the polluted plot to convert available C_org into new biomass was found in laboratory incubations with glucose-amended samples.     

Effects of biochar,earthworms, and litter addition on soil microbial activity and abundance in a temperate agricultural soil

Biochar application to arable soils could be effective for soil C sequestration and mitigation of greenhouse gas (GHG) emissions. Soil microorganisms and fauna are the major contributors to GHG emissions from soil, but their interactions with biochar are poorly understood. We investigated the effects of biochar and its interaction with earthworms on soil microbial activity, abundance, and community composition in an incubation experiment with an arable soil with and without N-rich litter addition. After 37 days of incubation, biochar significantly reduced CO₂ (up to 43 %) and N₂O (up to 42 %), as well as NH₄ ⁺-N and NO₃ ^?-N concentrations, compared to the control soils. Concurrently, in the treatments with litter, biochar increased microbial biomass and the soil microbial community composition shifted to higher fungal-to-bacterial ratios. Without litter, all microbial groups were positively affected by biochar × earthworm interactions suggesting better living conditions for soil microorganisms in biochar-containing cast aggregates after the earthworm gut passage. However, assimilation of biochar-C by earthworms was negligible, indicating no direct benefit for the earthworms from biochar uptake. Biochar strongly reduced the metabolic quotient qCO₂ and suppressed the degradation of native SOC, resulting in large negative priming effects (up to 68 %). We conclude that the biochar amendment altered microbial activity, abundance, and community composition, inducing a more efficient microbial community with reduced emissions of CO₂ and N₂O. Earthworms affected soil microorganisms only in the presence of biochar, highlighting the need for further research on the interactions of biochar with soil fauna.     

Influence of earthworm activity and rice straw application on the soil microbial community structure analyzed by PLFA pattern

The earthworm Pheretima hilgendorfi, one of the most common anecic species in Japan, abounds in soils with applied rice-straw residues. The influences of the worm’s activity and/or those of rice-straw application on the soil microbial community structure were studied using a microcosm approach. Low Humic Andosol was incubated with or without earthworms in a jar for a month after the following treatments: 1) no treatment, 2) chopped rice straw top-dressed on the soil and 3) rice straw incorporated into the soil. The soil NO₃^–-N level increased with the earthworms’ presence even in the soil without rice straw. The soil NH₄⁺-N level was by far the highest in the soil treated with worms and no rice straw. Amounts of total and bacterial PLFAs increased due to the earthworms’ presence when rice straw was incorporated. The proportion of unsaturated fatty acids increased in the earthworm treatment while the saturated fatty acids decreased, suggesting an increase in the Gram-negative bacterial proportion. The results of principal component analysis indicated that the rice straw and the earthworms affected the soil microbial community structure independently. Any direct influence of the PLFAs contained in the worms’ bodies and in the rice straw on the soil’s PLFA profile was thought to be small.     

Influence of irrigated agriculture on soil microbial diversity

Organic carbon (C), bacterial biomass and structural community diversity were measured in Southern Idaho soils with long term cropping histories. The soils tested were native sagebrush vegetation (NSB), irrigated moldboard plowed crops (IMP), irrigated conservation – chisel – tilled crops (ICT) and irrigated pasture systems (IP). Organic C concentration in soils decreased in the order NSB 0–5 cm > IP 0–30 cm = ICT 0–15 cm > IMP 0–30 cm > NSB 5–15 cm = NSB 15–30 cm. Active bacterial, fungal and microbial biomass correlated with soil C as measured by the Walkely Black method in positive curvilinear relationships (r² = 0.93, 0.80 and 0.76, respectively). Amplicon length heterogeneity (LH-PCR) DNA profiling was used to access the eubacterial diversity in all soils and at all depths. The Shannon–Weaver diversity index was used to measure the differences using the combined data from three hypervariable domains of the eubacterial 16S rRNA genes. Diversity was greatest in NSB 15–30 cm soil and lowest in the IMP soil. With the exception of IMP with the lowest diversity index, the samples highest in C (NSB 0–5 cm, IP 0–30 cm, ICT 0–15 cm) reflected lower diversity indices. However, these indices were not significantly different from each other. ICT and IP increase soil C and to some extent increase diversity relative to IMP. Since soil bacteria respond quickly to environmental changes, monitoring microbial communities may be one way to assess the impact of agricultural practices such as irrigation and tillage regimes.     

Discriminating the effects of agricultural land management practices on soil fungal communities

The structure of fungal communities was examined in soil subjected to 5 years of different agricultural land management and tomato production practices. Length heterogeneity polymerase chain reaction (LH-PCR) of fungal rDNA internal transcribed spacer-1 (ITS-1) regions was used to create genomic fingerprints of the soil fungal communities. Three years after initiation of land management practices, univariate analysis of genetic diversity failed to detect differences among soil fungal communities in plots managed organically, conventionally or maintained free of vegetation by continuous tillage (disk fallow). Genetic diversity was significantly higher in plots maintained as a perennial pasture grass (Paspalum notatum var Argentine bahiagrass) or as an undisturbed weed fallow. The composition of soil fungal communities within organic, pasture grass or disk fallow plots were separated into unique clusters by non-parametric multivariate analysis of their Bray-Curtis similarity matrices, computed from the relative abundance of ITS-1 amplicons, while the composition of communities within disk fallow and conventional plots could not be distinguished from each other. Diversity of soil fungal communities was significantly reduced following the cultivation of tomato in year four when compared to the diversity in plots where tomato was not cultivated. Divergence in the composition of soil fungal communities was observed following the cultivation of tomato under all land management regimes except organic, where communities continued to remained clustered based upon similarities among their ITS-1 amplicons. Divergence in the composition of fungal communities became more pronounced following two major hurricanes (Francis and Jeanne, September 2004) except for communities in the organic and pasture grass plots. Following the completion of a second tomato crop in year 5, genetic diversity and richness was similar under all land management regimes except the pasture grass, where it remained significantly higher. By contrast, following two consecutive years of tomato production, unique but mutually similar compositions of fungal communities were detected only in plots subjected to the organic land management regime. This was supported by observations that fungal communities were dominated by a 341 bp rDNA amplicon fragment in all land management regimes except the organic. Cloning and sequencing indicated that the 341 bp fragment generated by LH-PCR had a sequencing size of 343 bp, which was most closely related to Fusarium oxysporum. Thus, land management practices that disturb or disrupt soil fungal communities will significantly reduce their diversity. However, the composition of soil fungal communities is more strongly influenced by land management practices and communities within an organically management system were more resistant to anthropogenic and meteorological disturbances.