Titanium dioxide nanoparticles (TiO₂NP) are often released into the soil through repeated discharge of wastewater and repetitive applications as fertilizers. Adverse effects of a single pulse on soil bacterial communities have been widely studied, while the impact of repeated exposure is poorly understood. This study compared the impacts of single and repeated exposure scenarios on the soil bacterial community. The repeated exposure promoted the total bacterial biomass but reduced the community diversity and induced larger alterations in community composition compared to the single exposure. Regarding the dosing frequencies of repeated exposure, community divergence increased in initial dosing cycles, and community stability was re-established and remained in subsequent dosing cycles. According to the different tolerance to dosing frequencies, the dynamic response patterns of the featured OTUs and functional genes could be classified into four types: 1) promotion, 2) suppression-recovery-promotion, 3) promotion-suppression-stable, and 4) suppression. These results suggest that chronic exposure with repetitive low-dosing of nanoparticles induced a tendency towards larger alteration of both community composition and functioning than in case of application of a single pulse of the same dosage. This study brings new insight into understanding the compositional and predicted functional dynamics of the soil bacterial community in response to nanoparticles and identifies a data gap in realistic time-variable exposure testing.

Titanium dioxide nanoparticles (TiO₂NP) are increasingly released in soil ecosystems, while there is limited understanding of the impacts of TiO₂NP on soil bacterial communities. Here we investigated the effects of TiO₂NP on the taxonomic composition and functional profile of a soil bacterial community over a 60-day exposure period. In short-term exposure (1-day), contradictory effects on the taxonomic composition of soil bacterial communities were found after exposure to a low realistic environmental concentration of TiO₂NP at 1 mg/kg as compared to the effects induced by medium and high concentrations of TiO₂NP at 500 and 2000 mg/kg. After long-term exposure (60-day), the negative effects of TiO₂NP at the low concentration disappeared, and the inhibition by TiO₂NP of the abundance of core taxa was enhanced along with increasing exposure concentrations. However, although significant alterations were observed in the taxonomic composition over time and exposure concentrations, no significant change was observed in the community functional profile as well as enzyme activity after 60-day exposure, indicating that functional redundancy likely contributed to the bacterial community tolerance after the exposure to TiO₂NP. Our study highlighted the importance of assessing bacterial community compositional and functional responses in assessing the environmental risk of nanoparticles on soil ecosystems.