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, 2001). We added 2?��L radiolabelled Leu ([3H]Leu, 37?MBq?mL?1, 5.74?TBq?mmol?1, http://www.selleckchem.com/products/BIBF1120.html Amersham) combined with nonlabelled Leu to each tube, resulting in 275?nM Leu in the bacterial suspensions. The amount of Leu incorporated into extracted bacteria per hour and gram soil was used as a measure of bacterial growth. Fungal growth was assessed using the acetate incorporation into ergosterol method (Newell & Fallon, 1991) adapted for soil (Pennanen et al., 1998; B??th, 2001) with modifications (Rousk et al., 2009b), adding 1-[14C]acetic acid (sodium salt, 7.4?MBq?mL?1, 2.04?GBq?mmol?1, Amersham) combined with unlabelled sodium acetate resulting in a final acetate concentration of 220?��M in a soil slurry and having a 4-h incubation at 22?��C without light. Ergosterol was extracted, separated and quantified using HPLC equipped with a UV detector (282?nm). The ergosterol peak was collected and the amount of incorporated radioactivity was determined. The amount of acetate incorporated into fungal ergosterol was used as a measure of fungal growth. Respiration was determined from CO2 evolved from 3?g soil in 20-mL glass vials closed with crimp caps and incubated under dark conditions at 22?��C for 22�C24?h. The CO2 was determined using GC. Microbial biomass http://www.selleck.cn/products/carfilzomib-pr-171.html was estimated using the SIR method (Anderson & Domsch, 1978). Glucose?:?talcum (4?:?1; 6?mg?g?1) was added to the soil samples following the GC analyses. After 20?min, the atmosphere was purged of CO2 with pressurized air, after which http://www.selleckchem.com/products/Metformin-hydrochloride(Glucophage).html the vials were again closed with crimp caps, and incubated for 2�C3?h at 22?��C. The CO2 evolved was then determined. SIR respiration was converted to biomass using the relationship 1?mg?CO2?h?1 at 22?��C corresponds to 20?mg biomass C (recalculated from Anderson & Domsch, 1978), and that microbial biomass contains 45% C. The PLFA pattern was determined using 1?g of frozen soil according to Frosteg?rd et al. (1993) with modifications described by Nilsson et al. (2007). An internal standard (methyl nonadecanoate fatty acid, 19:0) was added before the methylation step. The PLFAs chosen to indicate bacterial biomass were i15:0, a15:0, i16:0, 16:1��9, 16:1��7c, 10Me16:0, cy17:0, i17:0, a17:0, 18:1��7 and cy19:0, while PLFA 18:2��6,9 was used to indicate fungal biomass (Frosteg?rd & B??th, 1996). Ergosterol, determined in the fungal growth estimations, was used as an alternative fungal biomass indicator. Soil pH was measured at a soil?:?water ratio of 1?:?2.5 (w/w). Air-dry soil (10?g,