Rapid estimation of microbial biomass in grassland soils by ultra-violet absorbance

A reliable and simple technique for estimating soil microbial biomass (SMB) is essential if the role of microbes in many soil processes is to be quantified. Conventional techniques are notoriously time-consuming and unreproducible. A technique was investigated that uses the UV absorbance at 280 nm of 0.5 M K₂SO₄ extracts of fumigated and unfumigated soils to estimate the concentrations of carbon, nitrogen and phosphorus in the SMB. The procedure is based on the fact that compounds released after chloroform fumigation from lysed microbial cells absorb in the near UV region. Using 29 UK permanent grassland soils, with a wide range of organic matter (2.9–8.0%) and clay contents (22–68%), it was demonstrated that the increase in UV absorbance at 280 nm after soil fumigation was strongly correlated with the SMB C (r=0.92), SMB N (r=0.90) and SMB P (r=0.89), as determined by conventional methods. The soils contained a wide range of SMB C (412–3412 μg g⁻¹ dry soil), N (57–346 μg g⁻¹ dry soil) and P (31–239 μg g⁻¹ dry soil) concentrations. It was thus confirmed that the UV absorbance technique described was a rapid, simple, precise and relatively inexpensive method of estimating soil microbial biomass.     

PH对红壤微生物生物量碳和生物量磷的影响

The impact of pH changes on microbial biomass carbon (Cmic) and microbial biomass phosphorus (Pmic) were examined for 3 red soils under citrus production with different lengths of cultivation. Soil pH significantly affected Cmic and Pmic. The Cmie and Pmic changes, as a function of soil pH, appeared to follow a normal distribution with the original soil pH value at the apex and as pH increased or decreased compared to the original soil pH, Cmic and Pmic declined. Moreover, there were critical pH values at both extremes (3.0 on the acidic side and 8.0 to 8.5 on the alkaline side), beyond which most of microorganisms could never survive. The effect of pH on Cmic and Pmic was also related to the original soil pH. The higher the original soil pH was, the less Cmic or Pmic were affected by pH change. It is suggested that soil microorganisms that grow in a soil environment with a more neutral soil pH range (i.e. pH 5.5-7.5) may have a greater tolerance to pH changes than those growing in more acidic or more alkaline soil pH conditions.     

Determination of soil microbial biomass phosphorus in acid red soils from southern China

A CHCl₃ fumigation and 0.03 M NH₄F-0.025 M HCl extraction procedure was used to measure microbial biomass P (P_mic) in 11 acid red soils (pH <6.0) from southern China and the results compared to those obtained by the commonly-used CHCl₃ fumigation and 0.5 M NaHCO₃ extraction method. Extraction with NH₄F-HCl was found to be more effective and accurate than NaHCO₃ extraction for detecting the increase of P from microbial biomass P following chloroform fumigation due to its higher efficiency in extracting both native labile phosphate and added phosphate (³²P) in the soils. This was confirmed by the recovery of ³²P from in situ ³²P-labeled soil microbial biomass following fumigation and extraction by the NH₄F-HCl solution. Soil microbial biomass P, measured by the NH₄F-HCl extraction method, was more comparable with soil microbial biomass C (with a more narrow C:P ratio range of 4.3 to 22.3 and a mean of 15.6 in the microbial biomass), than that obtained by NaHCO₃ solution (with a mean C:P ratio of 30.7 and a wide range of 14.9 to 48.9). K_p, the fraction of soil microbial biomass P extracted after CHCl₃ fumigation, by the NH₄F-HCl solution was 0.34. The amount of microbial biomass P determined (using K_p =0.34) was 3–400% (mean 131%) higher than that obtained by the NaHCO₃ extraction (using K_p =0.40) for the 11 red soils studied. The results suggest that the CHCl₃ fumigation and NH₄F-HCl extraction method is more reliable for measuring microbial biomass P than the NaHCO₃ extraction method in acid red soils.     

Quantifying microbial biomass phosphorus in acid soils

 This study aimed to validate the fumigation-extraction method for measuring microbial biomass P in acid soils. Extractions with the Olsen (0.5 M NaHCO₃, pH 8.5) and Bray-1 (0.03 M NH₄F–0.025 M HCl) extractants at two soil:solution ratios (1 : 20 and 1 : 4, w/v) were compared using eight acid soils (pH 3.6–5.9). The data indicated that the flushes (increases following CHCl₃-fumigation) of total P (P_t) and inorganic P (P_i) determined by Olsen extraction provided little useful information for estimating the amount of microbial biomass P in the soils. Using the Bray-1 extractant at a soil:solution ratio of 1 : 4, and analysing P_i instead of P_t, improves the reproducibility (statistical significance and CV) of the P flush in these soils. In all the approaches studied, the P_i flush determined using the Bray-1 extractant at 1 : 4 provided the best estimate of soil microbial biomass P. Furthermore, the recovery of cultured bacterial and fungal biomass P added to the soils and extracted using the Bray-1 extractant at 1 : 4 was relatively constant (24.1–36.7% and 15.7–25.7%, respectively) with only one exception, and showed no relationship with soil pH, indicating that it behaved differently from added P_i (recovery decreased from 86% at pH 4.6 to 13% at pH 3.6). Thus, correcting for the incomplete recovery of biomass P using added P_i is inappropriate for acid soils. Although microbial biomass P in soil is generally estimated using the P_i flush and a conversion factor (k _P) of 0.4, more reliable estimates require that k _P values are best determined independently for each soil. Received: 3 February 2000     

长期施肥对红壤性水稻土微生物生物量与活性的影响

土壤微生物及其活性是指示土壤增肥过程和土壤环境变化的灵敏指标.本文研究了红壤荒地开垦为水田后不同施肥制度定位施肥 16 年后水稻土的微生物生物量与活性特征,结果表明:经 16 年水稻耕植,不同施肥措施下土壤的微生物生物量和活性还处于较低水平.施肥制度显著影响了水稻土的微生物生物量 C 和基质诱导呼吸,但对基础呼吸的影响还不明显.只施用 N、K 肥对提高土壤微生物生物量和活性没有显著效果,在不施肥或施用化肥的基础上配合有机循环可以显著提高土壤微生物的生物量、代谢活性和微生物呼吸的温度敏感性,N、P、K 肥配合有机循环的施肥制度对提高土壤微生物生物量和代谢活性的作用最好.     

Estimating microbial biomass N in soils with and without living roots: Limitations of a pre-extraction step

Special net-closed soil containers were used in a pot experiment with low and high plant densities to give soil samples with and without roots. Soils from the containers were analysed either by the fumigation-extraction method or by a modified procedure starting with a pre-extraction and sieving step to remove plant roots from the samples. In the extracts NO ₃ ^- -N, NH ₄ ⁺ -N, organic N, and total N were measured. Microbial biomass N was calculated from the differences in total N in fumigated and unfumigated soils. Different plant densities had almost no influence on the values of the N compounds using either method. In soils with roots, significantly more organic N (and total N) was found by the fumigation-extraction method compared to soils without roots while no differences were obtained using pre-extractions and sieving. Though the organic N content in pre-extracts from soils with roots was significantly higher than from soils without roots, the NO ₃ ^- -N and NH ₄ ⁺ -N content was lower. Significant differences in biomass N in soils with and without roots were found only with the fumigation-extraction method. Biomass N levels calculated using the results after pre-extraction and sieving were about 50% lower than levels detected using fumigation-extraction alone. With the use of special net-closed soil containers, not only were soil samples produced with and without roots, but it was also possible to induce a rhizophere in the soils. A comparison of the two methods using these soils clearly demonstrated that the method used has profound influence on the final biomass N results. While higher biomass levels were found by fumigation-extraction in soils with roots, because root N becomes extractable after fumigation, the use of a pre-extraction and a sieving step may underestimate the total biomass N content due to the pre-extraction of microbial N (especially from rhizosphere microorganisms) from the sample. Nevertheless, pre-extraction and sieving followed by fumigation-extraction does seem to be the preferable method for biomass N measurement in comparative studies, because in most cases only minor errors will occur.     

Soil microbial biomass estimates in soils contaminated with metals

The validity of the chloroform fumigation-incubation procedure for measuring soil microbial biomass in field soils contaminated with metals (e.g. Cu, Ni, Zn, Cd) was assessed. The metal contamination was the result of past sewage sludge additions and the contaminated field soils now contain metals at about current maximum U.K. recommended levels. The decomposition of native soil biomass or microbial material added after fumigation was little affected by the presence of metals and it was concluded that fumigation-incubation is a reliable procedure for measuring biomass in soils contaminated with moderate amounts of metals. This conclusion was confirmed by direct microscopy: similar soil biomass estimates were obtained by both methods.     

添加生物质炭对红壤水稻土有机碳矿化和微生物生物量的影响

通过室内培育试验,研究了添加生物质炭对江西红壤水稻土有机碳矿化和微生物生物量碳、氮含量的影响。结果表明:红壤有机碳矿化速率在培育第2天达最大值后迅速降低,培养7天后下降缓慢并趋于平稳;添加生物质炭降低了土壤有机碳的矿化速率和累积矿化量,培养结束时,不加生物质炭的对照处理中有机碳的累积矿化量分别比添加0.5%和1.0%生物质炭的处理高10.0%和10.8%。此外,生物质炭的加入显著提高了土壤微生物生物量,添加0.5%生物质炭处理的土壤微生物生物量碳、氮含量分别比对照高111.5%～250.6%和11.6%～97.6%,添加1.0%生物质炭处理的土壤微生物生物量碳、氮含量分别比对照高58.9%～243.6%和55.9%～110.4%。相同处理中,干旱的水分条件下(40%田间持水量)微生物生物量要高于湿润的水分条件(70%田间持水量)。同时,添加0.5%和1.0%的生物质炭使土壤代谢熵分别降低2.4%和26.8%,微生物商减少了43.7%和31.7%。     

中国土壤微生物量的大小

The microbial biomass C, N and P of soils all over China were determined in this study to study their affecting factors. The results, about 100-417 mg C kg^-1 soil, 18-51 mg N kg^-1 soil and 4.4-27.3 mg P kg^-1 soil, showed the biomass C, N and P in linear relationship with the soil total organic C, toal N and soil organic P. The ratios of C: N and C:P, ranging from 5.6 to 9.6 and from 11.2 to 48.4 respectively, were affected by soil pH, texture, crop rotation, macroclimate, etc. The ratio of C:N in soil biomass increases gradually from the north to the south in China.     

Response of soil microbial biomass to straw incorporation

The response of the soil microbial biomass to cereal straw incorporation was assessed for three sites in Eastern Scotland. Increases in the C:N ratio of the biomass, due to straw inputs, were greater for soil with a history of straw incorporation than soil with no previous incorporation history. The biomass C:N ratio response to straw inputs was associated with increases in the fungal component of soil microbial respiration, fungal plate count (total and cellulolytic) and FDA-active lengths of fungal hyphae.     

Ratios between estimates of microbial biomass content and microbial activity in soils

The content levels and activities of the microbiota were estimated in topsoils and in one soil profile at agricultural and forest sites of the Bornhöved Lake district in northern Germany. Discrepancies between data achieved by fumigation-extraction (FE) and substrate-induced respiration (SIR), both used for the quantification of microbial biomass, were attributed to the composition of the microbial populations in the soils. In the topsoils, the active, glucose-responsive (SIR) versus the total, chloroform-sensitive microbial (FE) biomass decreased in the order; field maize monoculture (field-MM)>field crop rotation (field-CR) and dry grassland>beech forest. This ratio decreased within the soil profile of the beech forest from the litter horizon down to the topsoil. Differences between microbial biomass and activities suggested varying biomass-specific transformation intensities in the soils. The metabolic quotient (qCO₂), defined as the respiration rate per unit of biomass, indicates the efficiency in acquiring organic C and the intensity of C mineralization, while biomass-specific arginine-ammonification (arginine-ammonification rate related to microbial biomass content) seems to be dependent on N availability. The qCO₂, calculated on the basis of the total microbial biomass, decreased for the topsoils in the same order as did the ratio between the active, glucose-responsive microbial biomass to the total, chloroform-sensitive microbial biomass, in contrast to qCO₂ values based on the glucose-responsive microbial biomass, which did not. There was no difference between the levels of biomass-specific arginine-ammonification in topsoils of the fertilized field-CR, fertilized field-MM, fertilized dry grassland and eutric alder forest, but levels were lower in the beech forest, dystric alder forest, and unfertilized wet grassland topsoils. Ratios between values of different microbiological features are suggested to be more useful than microbiological features related to soil weight when evaluating microbial populations and microbially mediated processes in soils.     

Changes in microbial biomass and activity in soils amended with phenolic acids

The phenolic acids p-hydroxybenzoic, ferulic, caffeic and vanillic acid, were added to soil of the Countesswells series that had been fallow or carried crops of potatoes, peas or barley for two consecutive years. Changes in phenolic acid concentration, the soil biomass, the respiration rate, and soil amylase activity were measured over 28 days. All the phenolic acids were sorbed by the soils which was generally in the order caffeic > ferulic = vanillic > hydroxybenzoic acid. The phenolic acids stimulated soil respiration and increased the biomass as determined by the substrate-induced respiration method. but the fumigation method of biomass assessment gave anomalous results. The soil amylase activity was initially increased by phenolic acid amendments but soon decreased, and after 7 days was less than in non-amended soil although activity had increased again after 28 days. The rates of respiration and the total phenolic acid concentrations were similar to unamended controls after 28 days. The immediate respiration response, measured 1–6 h after amendment, indicated that caffeic acid gave the largest initial response of the phenolic tested, this being 55–72% of that given by glucose. Soil from the potato plot showed the highest immediate response to the phenolic acid amendments measured as a proportion of the respiration response to glucose. The findings suggest that some crops stimulate the growth of phenolic-acid degrading organisms.     

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.     

湘北红壤丘岗稻田土壤有机碳、养分及微生物生物量空间变异

通过对湘北典型红壤丘岗254个稻田耕层样(01～8.cm)进行分析,比较了微地形对稻田土壤有机碳、氮、磷和微生物生物量的影响。结果表明,丘岗底部稻田土壤有机碳、全氮、微生物生物量碳、微生物生物量氮、可溶性氮含量分别比丘岗中下部稻田高14.6%1、3.6%、24.6%、20.4%和95.8%,丘岗中下部稻田土壤Olsen-P含量比丘岗底部稻田高33.3%,差异均达极显著水平(P0.01)。不同部位稻田土壤全磷、微生物生物量磷含量和有效磷库(微生物生物量磷与Olsen-P之和)含量差异不显著。此外,丘岗底部稻田土壤碳磷比、微生物生物量碳磷比和微生物商比丘岗中下部稻田高12.7%,28.5%,8.2%,其差异达显著(P0.05)或极显著(P0.01)水平。但微生物生物量氮/全氮、微生物生物量磷/全磷、土壤碳氮比和微生物生物量碳氮比差异不显著。     

Fuzzy classification of microbial biomass and enzyme activities in grassland soils

Soil microbial properties are widely used as indicators of soil quality. The interpretation of soil microbial processes, however, is difficult because of their regional and seasonal heterogeneity as well as the lack of reference values. One possibility to overcome these limitations to apply the fuzzy set theory. This approach more realistically describes ecological systems because it considers natural ambiguity and complexity. The present study applies a fuzzy rule-based classification model to define soil quality based on soil microbial biomass, N-mineralisation, enzyme activity data (urease, xylanase, phosphatase, arylsulfatase) and soil organic matter. The data have been collected from different grassland sites in the European Union over a period of 20 years. The fuzzy model is based on a rule system derived from a training set using simulated annealing as an optimisation algorithm. For each variable, nine triangular fuzzy sets were defined for consideration as possible rule arguments. The model uses the t-norm for combination of arguments, product inference, the weighted sum as rule combination and the maximum method for defuzzification. The output is the assignment of membership of the object to a given soil quality class. The soil quality classes (very poor, poor, medium, high, very high) were defined by five heavy metal contamination levels (very high, high, medium, low, no). A predefined number of fuzzy rules were assessed using a simulated annealing algorithm. The fuzzy model was validated by a test file by assigning the soils to the quality class with the highest response value. The fuzzy model yielded an overall coincidence of 88.8% between observed and simulated results. The most sensitive index of soil quality was microbial biomass. N-mineralisation was a good indicator for the high-quality soils, while urease and arylsulfatase were important indicators for heavily contaminated, poor soil quality. Xylanase and phosphatase behaved ambivalently. Including soil organic carbon in the model decreased its effectiveness by 6.5%. We suggest that the presented fuzzy model based on soil microbial properties could be applied not only to soil degradation, upscaling and prediction, but also to judge the response of soils to environmental changes.     

Seasonal changes in microbial biomass and nutrient flush in forest soils

Microbial biomass and N, P, K, and Mg flushes were estimated in spring, summer, autumn, and winter samples of different forest soils. The microbial biomass showed significant seasonal fluctuations with an average distribution of 880±270 g C g^-1 soil in spring, 787±356 g C g^-1 soil in winter, 589±295 g C g^-1 soil in summer, and 560±318 g C g^-1 soil in autumn. The average annual concentrations of C, N, P, K, and Ca in the microbial biomass were 704, 106, 82, 69 and 10 g g^-1 soil, respectively. Microbial C represented between 0.5 and 2% of the organic soil C whereas the percentage of microbial N with respect to the total soil N was two-to threefold higher than that of C; the annual fluctuations in these percentages followed a similar trend to that of the microbial biomass. Microbial biomass was positively correlated with soil pH, moisture, organic C, and total N. The mean nutrient flush was 31, 15, 7, and 4 g g^-1 soil for N, K, P, and Mg, respectively, and except for K, the seasonal distribution was autumn spring winter summer. The average increase in available nutrient due to the mineralization of dead microbial cells was 240% for N, and 30, 26, and 14% for P, K, and Mg, respectively. There was a positive relationship between microbial biomass and the N, P, K, and Mg flushes. All the variables studied were significantly affected by the season, the type of soil, and the interaction between type of soil and season, but soil type often explained most of the variance.     

Turnover time of microbial biomass carbon in Japanese upland soils with different textures

We investigated the turnover time of microbial biomass-C in Japanese upland soils with various textures and examined the soil physicochemical properties influencing their turnover time. Samples from five different soil types (sand-dune regosol, light-colored Andosol, humic Andosol, brown forest soil, and dark red soil) were taken from upland concrete-frame plots in the experimental field of Chiba University. Each soil amended with [U -¹³C] glucose was incubated for 80 d at 25°C. Microbial biomass-C and -¹³C in soil were periodically determined by the fumigation-extraction method. The longest turnover time of microbial biomass-C was observed in the dark red soil (215 d) followed by the humic Andosol (134 d), brown forest soil (97 d), and light-colored Andosol (83 d) and the shortest in the sand-dune regosol (45 d). The turnover time of microbial biomass-C was significantly correlated with the value of soil clay (R: 0.917*), CEC (R: 0.921*), and macroaggregate (R: 0.907*) contents, but not with the total-C content. The amount of microbial biomass-C showed a close correlation with the turnover time of microbial biomass-C, suggesting that the turnover time of microbial biomass-C is an important factor influencing the accumulation of microbial biomass-C in soil.     

Effects of heavy metal accumulation in apple orchard soils on microbial biomass and microbial activities

Abstract

The effects of heavy metals (Cu, Pb, and As) accumulated in apple orchard surface soils on the microbial biomass, dehydrogenase activity, and soil respiration were investigated. The largest concentrations of total Cu, Pb, and As found in the soils used were 1,010, 926, and 166 mg kg^?1 soil, respectively. The amounts of microbial biomass C and N, expressed on a soil organic C and soil total N basis, respectively, were each negatively correlated with the amounts of total, 0.1 M HCI-extractable, and 0.1 M CaCl₂-extractable Cu as logarithmic functions, the correlation coefficient being lowest for the 0.1 M CaCl₂extractable Cu. Nevertheless, they were not correlated with the soil pH which was controlling the solubility of Cu in 0.1 M CaCl₂. The dehydrogenase activity expressed per unit of soil organic C was also negatively correlated with the amounts of total, 0.1 M HCI-extractable Cu, and 0.1 M CaCl₂-extractable Cu as logarithmic functions. However, the correlation coefficient was highest for the 0.1 M CaCl₂-extractable Cu. Although the soil respiration per unit of soil total organic C did not show any significant correlations with the total concentrations of heavy metals, it showed negative significant correlations with the amount of 0.1 M HCI-extractable Cu, and to a greater extent, with the amount of 0.1 M CaCl₂-extractable Cu. Both the dehydrogenase activity and respiration per unit of soil total organic C increased significantly with increasing soil pH. These results suggested that in apple orchard soils with heavy metal accumulation the microbial biomass was adversely affected by the slightly soluble Cu, whereas the microbial activities by the readily soluble Cu whose amount depended on the soil pH. The respiration per unit of microbial biomass C showed a positive significant correlation with the logarithmic concentration of total Cu. Furthermore, the contribution of fungi to substrate-induced respiration increased with increasing total Cu content in the soils.     

Soil microbial biomass and organic matter composition in soils under cultivation

To determine whether there is a relationship between the composition of soil organic matter and the activity of the soil microbial biomass, the composition of the organic matter in 12 typical arable soils in Northwest Germany was investigated by wet chemical analysis and CPMAS cross polarization magic angle spinning ¹³C-NMR spectroscopy. The data were correlated with the microbial biomass as estimated by substrate-induced respiration. A strong correlation between the microbial biomass and alkylic C compounds was observed (r=-0.960^***). Recalcitrant substances were enriched in this fraction, which were classified as humic acids according to the wet chemical procedure. The microbial decomposition of these humic acids is probably retarded, due to their chemical structure and/or physical bonding, when the soil microbial biomass activity is limited.