Application of synthetic herbicides is currently the most widely used and cost-effective methods to assist with revegetation programs. However, the effects of short-term application of herbicides such as Roundup®, acetic acid, BioWeed™ and Slasher® as compared with mulch, on soil microbial biomass and microbial diversity remain unknown. This study examined the effects of short-term herbicide application on soil microbial biomass, C:N ratio, and fungal and bacterial communities at months 2 and 8 following initiation of treatment application. No effects of treatments on soil pH, C:N and microbial biomass were found. No segregation among treatments in the community structure of bacteria and fungi was observed. However, the fungal phylum Basiodiomycota had one unidentified class, which was only found in the mulch treatment, suggesting the C quality in the mulch treatment may differ compared with the other treatments. The dry and hot conditions experienced throughout the study period may have resulted in fast degradation of the herbicides and may have minimised the impacts of the herbicides on microbial diversity and community structure. Given that the research was undertaken at a single site and over only a short time frame, the results should be extrapolated with caution. Herbicides may have greater impact with long-term use. Future research will need to assess the revegetation success of each treatment and determine if the observed change in Basidiomycota profile and C quality identified in this study becomes significant over the long-term. We hypothesise that mulching may be a preferred treatment to facilitate weed control in riparian zone revegetation. Copyright © 2019 Elsevier Ltd. All rights reserved.