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Soil microbiome responses to disturbance in arid ecosystems remains a critical knowledge gap. This study examines the soil microbiome in the north-east pastoral zone of South Australia, comparing the TGB Osborn Vegetation Reserve, established in 1925 with livestock and rabbit-proof fencing, to the adjacent grazed Koonamore Station, a merino sheep-grazing lease. Soil bacterial and fungal community compositions were analysed through amplicon sequencing, revealing that livestock grazing is associated with specific microbial abundance s and community structure, with distinct spatial patterns between bare soil areas and soil under perennial vegetation. Soils from open spaces in livestock grazing areas showed more disparity through reduced microbial abundances compared to soils ungrazed for 9 8 years. However, soils underneath shrub canopies in grazed areas showed increased abundances and taxonomic differences in comparison to ungrazed. Further research is needed to understand the functional consequences of these soil microbiome shifts on ecosystem function and services.