Real-time biodiversity monitoring should provide more resolved data to quantify shifts in ecological communities progressively altered by anthropogenic disturbances. Identifying biodiversity trends requires a rapid and efficient inventory method that enables the collection and delivery of high-resolution data within short intervals. Using aquatic environmental DNA (eDNA), we investigated spatiotemporal changes in fish and mammal communities along the Maroni River in French Guiana. We compared spatial biodiversity trends between two years, separated by a four-year interval, during which an increase in anthropogenic disturbances was observed. To evaluate biodiversity changes, we examined the impact of these disturbances on both taxonomic and functional diversity. Our findings revealed that, while the increase in disturbances did not result in major biodiversity decline, it continued to drive alterations in community taxonomic and functional richness. Communities underwent changes in their functional structure, with mammal communities experiencing a decline in extreme functional traits, while fish communities lost functional redundancy in generalist functions and experienced a reduction in extreme functional strategies. In a context of small-scale anthropogenic disturbances, these changes highlight the necessity of long-term, short-interval monitoring to capture rapid reorganisation of ecological communities under stress.