Polyethylene terephthalate microplastic fibers increase the release of extracellular antibiotic resistance genes during sewage sludge anaerobic digestion

•MFs increase the abundances of eARGs during sludge anaerobic digestion.•The increase of eARGs release by MFs has no significant selectivity.•The potential host number of eARGs also increased with MFs exposure.•Increased eARGs release could attribute to the upregulated type IV secretion system.•Increased eARGs may promote ARGs spread and should be considered in sludge handling. Microplastic fibers (MFs), as the most frequently detected microplastic shape in sewage sludge, have posed emerging concern for sludge treatment and disposal. However, the effect of MFs on antibiotic resistance genes (ARGs), especially extracellular ARGs (eARGs) during sludge treatment remains far from explicit. Therefore, this study investigated the potential impact of MFs on eARGs during sludge anaerobic digestion (AD), a commonly used sludge treatment method, through long-term operation. The qPCR results showed that both absolute and relative abundances of eARGs increased with the MFs exposure during sludge AD. The average absolute and relative abundances of eight tested eARGs in the AD reactor with the highest MFs dosage (170 items/gTS) were 1.70 and 2.15 times higher than those in the control AD reactor. The metagenomics results further comfirmed the increase of eARGs abundance during sludge anaerobic digestion after MFs exposure and the enhancement did not show significant selectivity. The identification of the potential hosts of eARGs suggested the host numbers of eARGs also increased with MFs exposure, which may suggest enhanced horizonal transformation as a result of increased eARGs. Further exploring the mechansims showed that the genes involved in type IV secretion system was upregulated after MFs exposure, suggesting the active release of eARGs was enhanced with MFs exposure. In contrast, the MFs may not affect the passive release of eARGs as its impact on cell membrance damage was insignificant. The enhanced eARGs in sludge AD process may further accelerate the transport of ARGs in environment, which should be considered during sludge treatment and disposal. [Display omitted]