Effects of Nano-Titanium Dioxide on the Horizontal Transfer of Antibiotic Resistance Genes in Microplastic Biofilms

Emerging pollutants such as microplastics in water environments readily accumulate microorganisms on their surfaces, forming biofilms and concentrating antibiotic-resistant bacteria (ARB) and antibiotic resistance genes (ARGs). Consequently, microplastic biofilms have attracted the attention of researchers. Horizontal gene transfer (HGT) of ARGs is one of the primary ways that bacteria acquire antibiotic resistance. Most studies focus on the effects of nanomaterials on suspended bacteria, but microplastic biofilms as hotspots for horizontal gene transfer also warrant significant investigation. This study primarily explored and compared the effects of nano-titanium dioxide on the conjugation transfer frequency of ARGs in suspended bacteria and microplastic biofilms. Nano-titanium dioxide could promote ARG conjugation in both suspended bacteria and microplastic biofilms, with a greater effect on the former. The mechanism involved nano-titanium dioxide promoting the production of reactive oxygen species (ROS) in suspended and biofilm bacteria, increasing the synthesis of outer membrane proteins, enhancing the cell membrane permeability, and elevating the expression levels of conjugation-related genes, thereby facilitating the conjugation transfer of ARGs. Biofilm bacteria, being heavily encased and protected by extracellular polymeric substances (EPS), exhibit greater resistance to external environmental pressure, resulting in the weaker impact of nano-titanium dioxide on biofilm bacteria compared to suspended bacteria. This study reveals the risk of ARG conjugation transfer within microplastic biofilms induced by nanomaterials, providing valuable insights into the risks of microplastic and antibiotic resistance dissemination in water environments.