As the human population grows, anthropogenic disturbances increasingly threaten natural environments, impacting biodiversity within ecosystems. Traditional bioindicators which rely on species presence and abundance, are valuable for monitoring environmental health. However, the gut microbiome, which can reflect the health of an organism, may serve as a novel bioindicator, particularly due to the rapid response of microbial communities to ecological changes. Spiders, abundant in the environment and known for inhabiting diverse ecological niches, are effective bioindicators and could potentially reflect environmental pressures through their gut microbiota. Here we investigated the gut microbiome of the Australian spider species, the Redback spider (Latrodectus hasselti) and the Black House spider (Badumna insignis), captured from various locations in Victoria to assess their potential as bioindicators of anthropogenic disturbance. Utilizing 16S rRNA sequencing, the taxonomic composition and diversity of the spider gut was analysed. The results revealed that the bacterial phyla Proteobacteria and Firmicutes dominated the gut of Redback spiders and Black House spiders, respectively. Furthermore, the Black House spiders exhibited a highly diverse and stable gut microbiome, while the Redback spiders showed significant variability between individuals. No correlation was observed between the spider gut composition and the individuals' geographic location, suggesting that other factors may influence the gut microbiome of these species. These findings indicate that the gut microbiome of Black House spiders may be a potential bioindicator for monitoring human-induced environmental impacts. This preliminary study establishes a foundation for future controlled research to identify the main drivers influencing the gut microbiome of arachnids and their response to anthropogenic disturbance.