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Sulfur deficiency arising due to intensive cultivation, use of sulfur free fertilizers and reduction in atmospheric sulfur depositions has become a major issue limiting crop production in many parts of the world. Elemental sulfur could be a good source of available S, but its slow oxidation is a problem for its efficient use as a sulfur fertilizer. Main objective of the study was to assess the effect of organic amendments (OA) and elemental sulfur (ES) on microbial activities, sulfur oxidation and availability in soil. A laboratory incubation experiment was carried out for a 56 days period using two sulfur deficient alkaline soils. Organic amendments (OA), i.e., farmyard manure (FYM), poultry litter (PL) and sugarcane filter cake (SF), were applied (1% <i>w</i>/<i>w</i>) with or without elemental sulfur (ES) at 50 mg kg⁻¹. Application of ES alone or in combination with OA significantly increasedCO₂-C evolution, microbial biomass, and enzyme activities in the soils, except dehydrogenase activity (DHA) which was not affected by ES application. Combined application of OA and ES had a more pronounced effect on microbial parameters compared to ES or OA applied alone. Ratios of dehydrogenase activity-to-microbial biomass C and arylsulfatase activity-to-microbial biomass C were high in ES+FYM and ES+SF treatments, respectively. Elemental sulfur got sufficiently oxidized resulting in significant improvement in plant available S. Relatively more ES was distributed into C-bonded-S than ester bonded-S. Increase in sulfur availability in ES+OA amended soils was the combined function of sulfur oxidation and mineralization processes through improved microbial activity.