Soil salinity is a widespread problem that affects crop productivity. We expect that saline soils also have altered microbial community structure, soil food webs and related soil properties. To test this, we sampled field soils across four farms in eastern North Dakota that host salinity gradients. We evaluated microbial biomass carbon, phospholipid fatty acid analysis and nematode counts in moderately saline and low saline soils. Additionally, we measured soil properties that represent potential food sources and habitat characteristics that influence microbial communities. We found higher microbial group abundance in moderately saline soils than in the lower saline soils. In contrast, we found lower nematode abundances in the moderately saline soils. We also observed increased labile carbon, nitrogen, phosphorus, and water content in the moderately saline soils. Based on our results, saline soils appear to have unique soil biological characteristics, which have implications for overall soil function along salinity gradients.