Distribution of extracellular and intracellular antibiotic resistance genes in sludge fractionated in terms of settleability.

The widespread proliferation of antibiotic resistance genes (ARGs) is a serious environmental and human health issue. Wastewater treatment facilities (WWTFs) are potential sources to spread ARGs to natural environment, for which, the presence state of ARGs in the sludge, as extracellular ones (eARGs) or intracellular ones (iARGs), along with the sludge settleability, are very important factors. The sludge settleability is closely associated with its floc size and density, bacterial activity, and the proportion of intact/damaged bacterial cells that aggregate together to form flocs for separation in the sedimentation process. It is reasonable to hypothesize that the distribution of eARGs and iARGs may differ with the sludge fractions of different settleability, a topic of great academic and practical significance requiring clarification. In this study, sludge samples from the aerobic contact tank of six household WWTFs were fractionated into fractions with different settling velocities: sludge of low settleability (LS), medium settleability (MS) and high settleability (HS); and the distribution of eARGs and iARGs in the obtained fractions for the widely detected tet G, tet M and sul 1 in water environment was evaluated based on the PMA-qPCR method, together with the evaluation for the well reported mobile genomic element intl 1 and total bacterial 16S rDNA. For the LS fractions, which contained more damaged bacterial cells, the distribution percentages of eARGs were generally higher than those of iARGs. For the HS fractions, which contained flocs with larger sizes formed by both intact and damaged bacterial cells, the relative abundances of ARGs and intl 1 were found apparently lower even if the presence percentages of eARGs were comparatively higher. It is thus inferable that sludge fractions of LS may possess higher transfer potential for ARGs and enhancing their settleability through optimization of the operation conditions is important for mitigating the proliferation of ARGs.