Activity and biomass of soil microorganisms at different depths

We measured microbial biomass C and soil organic C in soils from one grassland and two arable sites at depths of between 0 and 90 cm. The microbial biomass C content decreased from a maximum of 1147 (0–10 cm layer) to 24 g g^-1 soil (70–90 cm layer) at the grassland site, from 178 (acidic site) and 264 g g^-1 soil (neutral site) at 10–20 cm to values of between 13 and 12 g g^-1 soil (70–90 cm layer) at the two arable sites. No significant depth gradient was observed within the plough layer (0–30 cm depth) for biomass C and soil organic C contents. In general, the microbial biomass C to soil organic C ratio decreased with depth from a maximum of between 1.4 and 2.6% to a minimum of between 0.5 and 0.7% at 70–90 cm in the three soils. Over a 24-week incubation period at 25°C, we examined the survival of microbial biomass in our three soils at depths of between 0 and 90 cm without external substrate. At the end of the incubation experiment, the contents of microbial biomass C at 0–30 cm were significantly lower than the initial values. At depths of between 30 and 90 cm, the microbial biomass C content showed no significant decline in any of the four soils and remained constant up to the end of the experiment. On average, 5.8% of soil organic C was mineralized at 0–30 cm in the three soils and 4.8% at 30–90 cm. Generally, the metabolic quotient qCO₂ values increased with depth and were especially large at 70–90 cm in depth.     

The significance of microbial biomass sulphur in soil

The soil microbial biomass S fraction of total organic S in soil is considered to be relatively labile and the most active S pool for S turnover in soil. Its significance has been demonstrated in studies of S deficiency in agronomic situations and in those of S pollution from high atmospheric inputs. The utility of the CHCl₃ fumigation-extraction technique for the measurement of microbial S has been proved for a range of soils and conditions. The various methodologies currently available are discussed, including the need for determination of the conversion (K _s) factor. Microbial S values, summarized from the available literature, ranged from 3 to 300 g S g^-1 dry weight soil. They were generally greater in grassland than in arable systems, though the greatest values were obtained in the few examples from forest and peatland soil systems. Microbial S values showed direct relationships with both microbial C and with total soil organic S. Again, there were significant differences between arable and grassland systems. The effect of factors such as organic and inorganic inputs as well as soil physical conditions on microbial S are described. Microbial S turnover rates were estimated from seasonal, ³⁵S-labelling and modelling studies. These rates varied between an approximately annual turnover rate in undisturbed soils up to 80 year^-1 following the addition of readily available substrates. Prospective future research areas are also outlined.     

Effect of cultivation on microbial carbon and nitrogen in dry tropical forest soil

Summary Fifteen- and forty-year-old cropfields developed from a dry tropical forest were examined for soil organic C and total N and soil microbial C and N. The 15-year-old field had never been manured while the 40-year-old field had been fertilized with farmyard manure every year. The native forest soil was also examined. The results indicated that the native forest soil lost about 57% and 62% organic C and total N, respectively, in the 0–10 cm layer after 15 years of cultivation. The microbial C and N contents of the forest soil were greater than those of the cultivated soils. Application of farmyard manure increased the biomass-C and -N levels in the cultivated soil but the values were still markedly lower than in the forest soil. There was an appreciable seasonal variation in biomass C and N, the values being highest in summer and lowest in the rainy season. During an annual cycle, biomass-C contents varied from 180 to 727 g g^–1 and N from 20 to 80 g g^–1 dry soil, and both were linearly related. Microbial biomass C represented 1.6%–3.6% of total soil organic C and microbial biomass N represented 1.7% 1–4.4% of soil organic N.     

Influence of soil properties on microbial C,N, and P in dry tropical ecosystems

Summary Relationships between soil physicochemical characteristics and soil microbial C, N, and P in Indian dry tropical ecosystems are discussed. The major ecosystem studies were on forest, savanna, cropped fields, and mine spoils. The highest microbial C, N, and P levels were recorded from the mixed forest and the lowest levels in 5-year-old mine spoil. Across the sites, microbial C ranged from 226 to 643 g g^-1, microbial N from 19 to 71 g g^-1, and microbial P from 9 to 28 g g^-1 soil. The proportion of soil organic C contained in the microbial biomass ranged from 2.2 to 5.0%. The microbial C: N ratio in these soils ranged from 7.4. to 13.1 and the microbial C: P ratio from 16.6 to 30.6. The concentrations of microbial C, N, and P were correlated with several soil properties and among themselves. The soil properties, in various linear combinations, explained 90–99% of the variability in the microbial nutrients. Grazing of the savanna had some effect on the level of microbial biomass, and as the mine spoil aged, the level of microbial C, N, and P also increased.     

Application of the selective inhibition method to determine bacterial: fungal ratios in three beechwood soils rich in carbon — Optimization of inhibitor concentrations

Bacterial and fungal contributions to microbial respiration in three beechwood soils rich in C (two basalt soils and one limestone soil) were investigated by using streptomycin and cycloheximide to inhibit substrate-induced respiration after glucose (8000 g g^-1), N, and P addition to soil samples. The inhibitors were added as solutions (2000, 8000, and 16000 g g^-1) and the reduction in substrate-induced respiration after separate and combined inhibitor addition was measured in an automated electrolytic microrespirometer. Bacterial and fungal contributions to microbial respiration were calculated using the interval 6–10 h after inhibitor application. The microbial biomas was smaller in the two basalt soils (Oberhang and Mittelhang) than in the limestone soil (Unterhang). In the presence of both inhibitors, microbial respiration was inhibited by a maximum of 45, 45, and 25% in the two basalt soils and the limestone soil, respectively. Inhibition of microbial respiration was at a maximum at streptomycin and cycloheximide concentrations of 16000 g g^-1. The inhibitor additivity ratio approached 1.0 even at high inhibitor concentrations, indicating high inhibitor selectivity. Calculated prokaryote: eukaryote ratios indicated lower bacterial contributions to the microbial biomass in the Mettelhang (0.74) and Unterhang (0.73) than in the Oberhang (0.88) soil.     

Spatial changes of soil fungal and bacterial biomass from a sub-alpine coniferous forest to grassland in a humid, sub-tropical region

 Fungal and bacterial biomass were determined across a gradient from a forest to grassland in a sub-alpine region in central Taiwan. The respiration-inhibition and ergosterol methods for the evaluation of the microbial biomass were compared. Soil fungal and bacterial biomass both significantly decreased (P<0.05) with the shift of vegetation from forest to grassland. Fungal and bacterial respiration rates (evolved CO₂) were, respectively, 89.1 μl CO₂ g^–1 soil h^–1 and 55.1 μl CO₂ g^–1 soil h^–1 in the forest and 36.7 μl CO₂ g^–1 soil h^–1 and 35.7 μl CO₂ g^–1 soil h^–1 in the grassland surface soils (0–10 cm). The fungal ergosterol content in the surface soil decreased from the forest zone (108 μg g^–1) to the grassland zone (15.9 μg g^–1). A good correlation (R ²=0.90) was exhibited between the soil fungal ergosterol content and soil fungal CO₂ production (respiration) for all sampling sites. For the forest and grassland soil profiles, microbial biomass (respiration and ergosterol) declined dramatically with depth, ten- to 100-fold from the surface organic horizon to the deepest mineral horizon. With respect to fungal to bacterial ratios for the surface soil (0–10 cm), the forest zone had a significantly (P<0.05) higher ratio (1.65) than the grassland zone (1.05). However, there was no fungal to bacterial ratio trend from the surface horizon to the deeper mineral horizons of the soil profiles. Received: 30 March 2000     

Dynamics of soil biomass C,N, and P in a dry tropical forest in India

Summary Three dry tropical forest soils along a topographic sequence were examined to determine the seasonal dynamics of microbial C, N, and P. The lowest microbial biomass was found in forest soils at the foot of the hill followed by midslope forest soils. The hilltop soil, which had the most fine particles, water-holding capacity, organic C, and total N, reflected the presence of greater amounts of microbial C, N, and P. Mean annual microbial C, N, and P ranges were 466–662, 48–72 to 21–30 g g^-1, respectively. The seasonal pattern of microbial biomass, C, N, and P was similar at all sites, the values being greatest during the dry season and lowest during the wet season. The seasonal values for microbial biomass C, N, and P were positively correlated with each other and a negative correlation was found between microbial biomass and the fine root mass in these forest soils.     

Carbon and nitrogen relationships in the microbial biomass of soils in beech (Fagus sylvatica L.) forests

Soils from 38 German forest sites, dominated by beech trees (Fagus sylvatica L.) were sampled to a depth of about 10 cm after careful removal of overlying organic layers. Microbial biomass N and C were measured by fumigation-extraction. The pH of the soils varied between 3.5 and 8.3, covering a wide range of cation exchange capacity, organic C, total N, and soil C:N values. Maximum biomass C and biomass N contents were 2116 g C m^-2 and 347 g N m^-2, while minimum contents were 317 and 30 g m^-2, respectively. Microbial biomass N and C were closely correlated. Large variations in microbial biomass C:N ratios were observed (between 5.4 and 17.3, mean 7.7), indicating that no simple relationship exists between these two parameters. The frequency distribution of the parameters for C and N availability to the microflora divided the soils into two subgroups (with the exception of one soil): (1) microbial: organic C>12 mg g^-1, microbial:total N>28 mg g^-1 (n=23), a group with high C and N availability, and (2) microbial:organic C12 mg g^-1, microbial:total N28 mg g^-1 (n=14), a group with low C and N availability. With the exception of a periodically waterlogged soil, the pH of all soils belonging to subgroup 2 was below 5.0 and the soil C:N ratios were comparatively high. Within these two subgroups no significant correlation between the microbial C:N ratio and soil pH or any other parameter measured was found. The data suggest that above a certain threshold (pH 5.0) microbial C:N values vary within a very small range over a wide range of pH values. Below this threshold, in contrast, the range of microbial C:N values becomes very large.     

Wood-ash fertilization and fire treatments in a Scots pine forest stand: Effects on the organic layer,microbial biomass,and microbial activity

We studied the reactions of humus layer (F/H) microbial respiratory activity, microbial biomass C, and the fungal biomass, measured as the soil ergosterol content, to the application of three levels of wood ash (1000, 2500, and 5000 kg ha^-1) and to fire treatment in a Scots pine (Pinus sylvestris L.) stand. Physicochemical measurements (pH, organic matter content, extractable and total C content, NH ₄ ⁺ and total N content, cation-exchange capacity, base saturation) showed similarity between the fire-treated plots and those treated with the lowest dose of wood ash (1000 kg ha^-1). The ash application did not change the level of microbial biomass C or fungal ergosterol when compared to the control, being around 7500 and 350 g g^-1 organic matter for the biomass C and ergosterol, respectively. The fire treatment lowered the values of both biomass measurements to about half that of the control values. The fire treatment caused a sevenfold fall in the respiration rate of fieldmoist soil to 1.8 l h^-1 g^-1 organic matter compared to the values of the control or ash treatments. However, in the same soils adjusted to a water-holding capacity of 60%, the differences between the fire treatment and the control were diminished, and the ash-fertilized plots were characterized by a higher respiration rate compared to the control plots. The glucose-induced respiration reacted in the same way as the water-adjusted soil respiration. The metabolic quotient, qCO₂, gradually increased from the control level with increasing applications of ash, reaching a maximum in the fire treatment. Nitrification was not observed in the treatment plots.     

Interactions between 14C-labelled atrazine and the soil microbial biomass in relation to herbicide degradation

A laboratory incubation experiment was set up to determine the effects of atrazine herbicide on the size and activity of the soil microbial biomass. This experiment was of a factorial design (0, 5, and 50 g g^–1 soil of non-labelled atrazine and 6.6×10³ Bq g^–1 soil of ¹⁴C-labelled atrazine) x (0, 20, and 100 g g^–1 soil of urea-N) x (pasture or arable soil with a previous history of atrazine application). Microbial biomass, measured by substrate-induced respiration and the fumigation-incubation method, basal respiration, incorporation of ¹⁴C into the microbial biomass, degradation of atrazine, and ¹⁴C remaining in soil were monitored over 81 days. The amount of microbial biomass was unaffected by atrazine although atrazine caused a significant enhancement of CO₂ release in the non-fumigated controls. Generally, the amounts of atrazine incorporated into the microbial biomass were negligible, indicating that microbial incorporation of C from atrazine is not an important mechanism of herbicide breakdown. Depending on the type of soil and the rate of atrazine application, 18–65% of atrazine was degraded by the end of the experiment. Although the pasture soil had twice the amount of microbial biomass as the arable soil, and the addition of urea approximately doubled the microbial biomass, this did not significantly enhance the degradation of atrazine. This suggests that degradation of atrazine is largely independent of the size of the microbial biomass and suggests that other factors (e.g., solubility, chemical hydrolysis) regulate atrazine breakdown. A separate experiment conducted to determine total amounts of ¹⁴C-labelled atrazine converted into CO₂ by pasture and arable soils showed that less than 25% of the added ¹⁴C-labelled atrazine was oxidised to ¹⁴CO₂ during a 15-week period. The rate of degradation was significantly greater in the arable soil at 24%, compared to 18% in the pasture soil. This indicates that soil microbes with previous exposure to atrazine can degrade the applied atrazine at a faster rate.     

Identification of ergosterol in vesicular-arbuscular mycorrhizae

Summary Ergosta-5,7,22-tri-3-enol (ergosterol) was identified by gas chromatography-mass spectrometry in roots of berseem (Trifolium alexandrinum L., cv. Landsorte) and sweet corn (Zea mays L., cv. Honeycomb-F1) infected with the vesicular-arbuscular mycorrhizal (VAM) fungus Glomus intraradices. The fungal-derived compound ergosterol was determined quantitatively in root extracts using reverse-phase high performance liquid chromatography. The concentrations of ergosterol in VAM-infected roots reached 72 g^-1 dry material in berseem and 52 g^-1 in sweet corn after 80 days of growth, whereas concentrations in non-infected roots remained below 8 g^-1 dry weight. Additionally, phytosterols such as -sitosterol, campesterol, and stigmasterol were detected in both infected and non-infected roots. Ergosterol, as a characteristic fungal substance, is proposed as an indicator of fungal biomass in the early stages of VAM infection.     

Influence of soil properties on microbial populations, activity and biomass in humid subtropical mountainous ecosystems of India

 Microbial populations, biomass, soil respiration and enzyme activities were determined in slightly acid organic soils of major mountainous humid subtropical terrestrial ecosystems, along a soil fertility gradient, in order to evaluate the influence of soil properties on microbial populations, activity and biomass and to understand the dynamics of the microbial biomass in degraded ecosystems and mature forest. Although the population of fungi was highest in the undisturbed forest (Sacred Grove), soil respiration was lowest in the 7-year-old regrowth and in natural grassland (approximately 373 μg g^–1 h^–1). Dehydrogenase and urease activities were high in "jhum" fallow, and among the forest stands they were highest in the 7-year-old regrowth. Microbial biomass C (MBC) depended mainly on the organic C status of the soil. The MBC values were generally higher in mature forest than in natural grassland, 1-year-old jhum fallow and the 4-year-old alder plantation. The MBC values obtained by the chloroform-fumigation-incubation technique (330–1656 μg g^–1) did not vary significantly from those obtained by the chloroform-fumigation-extraction technique (408–1684 μg g^–1), however, the values correlated positively (P<0.001). The enzyme activities, soil respiration, bacterial and fungal populations and microbial biomass was greatly influenced by several soil properties, particularly the levels of nutrients. The soil nutrient status, microbial populations, soil respiration and dehydrogenase activity were greater in Sacred Grove, while urease activity was greater in grassland. Received: 14 October 1998     

The influence of tillage on the proportion of organic carbon and nitrogen in the microbial biomass of medium-textured soils in a humid climate

Summary Microbial biomass C and N respond rapidly to changes in tillage and soil management. The ratio of biomass C to total organic C and the ratio of mineral N flush to total N were determined in the surface layer (0–5 cm) of low-clay (8–10%), fine sandy loam, Podzolic soils subjected to a range of reduced tillage (direct drilling, chisel ploughing, shallow tillage) experiments of 3–5 years' duration. Organic matter dynamics in the tillage experiments were compared to long-term conditions in several grassland sites established on the same soil type for 10–40 years. Microbial biomass C levels in the grassland soils, reduced tillage, and mouldboard ploughing treatments were 561, 250, and 155 g g^-1 soil, respectively. In all the systems, microbial biomass C was related to organic C (r=0.86), while the mineral N flush was related to total N (r=0.84). The average proportion of organic C in the biomass of the reduced tillage soils (1.2) was higher than in the ploughed soils (0.8) but similar to that in the grassland soils (1.3). Reduced tillage increased the average ratio of mineral N flush to total soil N to 1.9, compared to 1.3 in the ploughed soils. The same ratio was 1.8 in the grassland soils. Regression analysis of microbial biomass C and percent organic C in the microbial biomass showed a steeper slope for the tillage soils than the grassland sites, indicating that reduced tillage increased the microbial biomass level per unit soil organic C. The proportion of organic matter in the microbial biomass suggests a shift in organic matter equilibrium in the reduced tillage soils towards a rapid, tillage-induced, accumulation of organic matter in the surface layer.     

The spatial distribution of soil microbial biomass in a permanent row crop

The effects of soil texture (silt loam or sandy loam) and cultivation practice (green manure) on the size and spatial distribution of the microbial biomass and its metabolic quotient were investigated in soils planted with a permanent row crop of hops (Humulus lupulus). The soil both between and in the plant rows was sampled at three different depths (0–10, 10–20, and 20–30 cm). The silt loam had a higher overall microbial biomass C concentration (260 g g^-1) than the sandy loam (185 g g^-1), whereas the sandy loam had a higher (3.1 g CO₂-C mg^-1 microbial Ch^-1) metabolic quotient than the silt loam (2.6 g CO₂-C mg^-1 microbial C h^-1), on average over depth (0–30 cm) and over all treatments. There was a sharp decrease in the microbial biomass with increasing depth for all plots. However, this was more pronounced in the silt loam than in the sandy loam. There was no distinct influence of sampling depth on the metabolic quotient. The microbial biomass was considerably higher in the rows than between the rows, especially in the silt loam plots. There was no significant difference between plots without green manure and plots with green manure for either the microbial biomass or the metabolic quotient.     

Examination of microbial biomass in beech forest moder profiles

Microbial biomass C, ATP, and substrate-induced respiration were measured in the organic layers and the mineral A horizon of three beech forest soils with moder humus differing in Ca and Mg supply. Analyses of variance showed that horizon-specific differences explained most of the variance in the three microbial parameters. All three were significantly interrelated, with Spearman rank correlation coefficients of between 0.86 and 0.93. However, differences in the decline of these parameters with depth led to horizon-specific differences in their ratios. Thus, the ratios were not markedly interrelated. The mean ATP: microbial C ratio was 5.2 mol ATP g^-1 C in the L 2 layer, 19.5 in the F layers, and 9.6 in the H and A horizons. The ratio of substrate-induced respiration to microbial C varied between 39.3 and 82.2 O₂h^-1 g^-1 C in the F1 layers and between 5.3 and 32.1 l in the other layers. It is concluded that the use of different parameters can help to analyze both horizonand site-specific differences in microbial performance.     

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.     

Effect of residue placement and chemical fertilizer on soil microbial biomass under tropical dryland cultivation

Four treatments (control, chemical fertilizer, wheat straw, and wheat straw+fertilizer) were established on the dryland experimental farm of the Institute of Agricultural Sciences, Banaras Hindu University. Organic in C in the different treatments ranged from 0.69 to 0.93%, total N from 0.08 to 0.11%, and total P from 0.018 to 0.021. The application of straw significantly increased the soil water-holding capacity. The maximum effect on the microbial biomass was realized with the straw+fertilizer treatment, followed by straw and then by the fertilizer treatment. During the study microbial biomass C ranged from 144 to 491 g g^-1 dry soil, biomass N from 14.6 to 50.1 g g^-1, and biomass P from 7.2 to 17.6 g g^-1 soil. Microbial biomass C, N and P represented 3.2–4.6% of total C, 2.6–3.8% of total N, and 5.8–8.2% of total P in the soil, respectively, in all cases the highest proportion occurred in the straw+fertilizer treatment and the lowest in the control. Microbial biomass C, N, and P were positively correlated with each other. Microbial biomass C and N increased by 77% in straw+fertilizer-treated plots relative to the control. The increase in microbial biomass P in the straw+fertilizer treatment over the control was 81%. The increase in the microbial biomass is expected to enhance nutrient availability in the soil, as the microbial biomass acts both as a sink and a source of plant nutrients.     

Effects of farmyard manure and inorganic fertilizer on the dynamics of soil microbial biomass in a tropical dryland agroecosystem

Changes in the soil microbial biomass following applications of farmyard manure and inorganic fertilizer, alone and in combination, were studied for two annual cycles in a rice-lentil crop sequence grown under rainfed tropical dryland conditions. During the two annual cycles the microbial biomass C range (g g^-1) was 146–241 (x = 204), 191–301 (245), 244–382 (305), and 294–440 (365) in control, fertilizer, manure and manure+fertilizer plots, respectively. The corresponding ranges for microbial biomass N (g g^-1) were 16.5–21.0 (19.5), 20.4–38.2 (26.0), 23.0–34.6 (27.0) and 26.2–42.4 (33.3), and for microbial biomass P (g g^-1) 4.4–8.2 (7.0) 6.0–11.2 (9.6), 11.2–22.0 (17.0), and 10.0–25.4 (18.3). The maximum increase in the microbial biomass, due to these inputs was observed under the manure+fertilizer treatment followed, in decreasing order, by manure alone and fertilizer alone. Within individual crop periods the levels of microbial biomass decreased sharply from the seedling to the flowering stage and then increased slightly with crop maturity. The maximum levels of microbial biomass C and P were observed during the summer fallow. The maximum accumulation of microbial biomass N occurred in the early rainy season, immediately after the soil amendments. Microbial biomass C, N, and P were positively related to each other throughout the annual cycle.     

The use of phospholipid fatty acid analysis to estimate bacterial and fungal biomass in soil

The cell content of 12 bacterial phospholipid fatty acids (PLFA) was determined in bacteria extracted from soil by homogenization/centrifugation. The bacteria were enumerated using acridine orange direct counts. An average of 1.40×10^-17 mol bacterial PLFA cell^-1 was found in bacteria extracted from 15 soils covering a wide range of pH and organic matter contents. With this factor, the bacterial biomass based on PLFA analyses of whole soil samples was calculated as 1.0–4.8 mg bacterial C g^-1 soil C. The corresponding range based on microscopical counts was 0.3–3.0 mg bacterial C g^-1 soil C. The recovery of bacteria from the soils using homogenization/centrifugation was 2.6–16% (mean 8.7%) measured by PLFA analysis, and 12–61% (mean 26%) measured as microscopical counts. The soil content of the PLFA 18:26 was correlated with the ergosterol content (r=0.92), which supports the use of this PLFA as an indicator of fungal biomass. The ratio 18:26 to bacterial PLFA is therefore suggested as an index of the fungal:bacterial biomass ratio in soil. An advantage with the method based on PLFA analyses is that the same technique and even the same sample is used to determine both fungi and bacteria. The fungal:bacterial biomass ratio calculated in this way was positively correlated with the organic matter content of the soils (r=0.94).     

Does biochar interfere with standard methods for determining soil microbial biomass and phenotypic community structure?

Due to its high sorption affinity for organic compounds, biochar may interfere with extraction procedures involving such compounds used for microbially-related assays commonly applied to soils. Here we assessed the impact of two biochars (derived from pine bark and produced at 300 and 600 °C) at three concentrations (0, 12.5, and 50 g kg⁻¹) in three distinct arable soils with contrasting textural classes (loamy sand, sandy loam, and clay) on the determination of soil microbial biomass C by fumigation–extraction, fungal biomass by ergosterol analysis, and microbial community structure as defined by phospholipid fatty acid (PLFA) profiling. Biochar did not affect the apparent concentration of soil microbial biomass C and had no significant impact on apparent PLFA profiles. By contrast, the apparent extraction efficiency of ergosterol was affected dependent on soil type, biochar production temperature, and biochar concentration. Nonetheless, ergosterol contents of biochar-amended soils can be accurately estimated by correcting for reduced recovery using an ergosterol spike.