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Nutrient availability is an important indicator of nutrient balance and affects soil microbial processes and functions. However, few studies have explored the effect of short-term application of mineral phosphorus (P) on grassland soils. This study evaluated the effect of mineral P fertilization on soil microbial biomass, activity and community composition in the soil of timothy (Phleum pratense L.) swards at two sites (established in 2010) in eastern Canada with P fertilization rates of 0, 20, and 40 kg P ha⁻¹. Soil samples were collected at the top 10 cm three times in 2013 and 2014. Results revealed that P application has significantly increased soil Mehlich-3P and microbial biomass P at both sites tested, but soil microbial biomass C and N were not affected. Phosphorus application has increased dehydrogenase activity, but it had no significant effect on alkaline phosphomonoesterase activity. The total soil bacterial …