Effect of grazing intensity on protozoan community, microbial biomass, and enzyme activity in an alpine meadow on the Tibetan Plateau

The alpine meadow has received mounting attention due to its degradation resulting from overgrazing on the Tibetan Plateau. However, belowground biotic characteristics under varied grazing stresses in this ecosystem are poorly understood. Here, the responses of soil protozoan abundance, community composition, microbial biomass, and enzyme activity to five grazing patterns including (1) artificial grassland without grazing (AG), (2) winter grazing (WG), (3) grazing for 7 months within a fence (GF), (4) continuous grazing for a whole year (CG), and (5) natural heavy grazing (HG) were investigated for two continuous years. Soil protozoan community composition was investigated using the most possible number (MPN) method, and soil microbial biomass and enzyme activity were analyzed using chloroform fumigation extraction and substrate utilization methods, respectively. Multivariate statistical analysis, the analysis of variance (ANOVA), multiple comparisons, and correlation analysis were together performed. The WG treatment had the highest abundance of total protozoa (2342-2524 cell g(-1)). Compared with AG treatment, HG treatment significantly reduced the abundance of soil total, flagellate and ciliate protozoa, and protease activities in 2012 and 2013. Significantly, lower soil microbial biomass nitrogen (MBN) was also observed in the HG (6.60 and 14.6 mg N kg(-1)) than those in other four treatments (22.3-82.9 mg N kg(-1)) both in 2012 and 2013, whereas significantly higher microbial biomass carbon (MBC) was observed in HG than that in AG treatment in 2012. Moreover, significantly positive correlations were detected between the abundance of soil protozoa and soil moisture, pH, organic C, total N, and MBN. Our results indicated that soil protozoa showed a negative response to increasing grazing intensities and therefore, suggesting that aboveground grazing practices also exerted strong impact on belowground protozoa, not only on soil microbial characteristics. Soil protozoan community composition was apparently different between the HG treatment and other four grazing patterns and was potentially impacted by altered soil properties and MBC and/or MBN. Our results suggested that moderate grazing may sustain better belowground biotic diversity and ecosystem functioning in this alpine meadow on the Tibetan Plateau.