The pig slurry (PS) application as an organic fertilizer in Brazilian soils occurs for many years and the effects of this application are known, but in this study our objective was to evaluate the microbiological attributes and microbial diversity of the soil after the interruption of the PS application. The PS rates were 0, 30 and 60 m(3) ha(-1) yr(-1) and in 2016 the plots were divided and the PS application was stopped in half of the plot, generating the factorial scheme 2 x 2, being two PS rates (30 and 60 m(3) ha(-1) yr(-1)) and two application conditions (continuous or interrupted). Two soil samples were taken at 5 and 12 mo after interruption. The PS interruption reduced the microbial biomass C (MBC) by 5% and 14% after 5 and 12 mo, respectively. Microbial biomass N (MBN) was reduced too by 32% and 29% due to the PS interruption. The soil basal respiration (SBR) decreased when PS application was interrupted. The metabolic quotient (qCO(2)) did not change with the PS interruption but was high in the 60 m(3) ha(-1) yr(-1). The bacteria and archaea genetic profiles showed similarity above 69% between continuous and interrupted applications. The bacterial diversity and richness index were higher than those found for fungal and archaeal communities. In contrast, the interruption had a strong effect on the soil fungi, which had diversity reduced due to the increase of the species dominance. The PS interruption up to 12 mo has an effect on the biomass, diversity, and activity of the microorganisms.