Gut digestion of earthworms significantly attenuates cell-free and -associated antibiotic resistance genes in excess activated sludge by affecting bacterial profiles

Vermicomposting can significantly attenuate antibiotic resistance genes (ARGs) in the excess activated sludge (EAS). However, the effect of earthworms, especially the effect of gut digestion as a critical step in the vermicomposting process, remains unclarified. The purpose of this study was to investigate the response of ARGs (cell-free and -associated) in EAS to gut digestion of earthworms and to clarify the possible mechanism from the viewpoint of bacterial community through quantitative polymer chain reaction (q-PCR) and high throughput sequencing. Compared to the initial sludge, the earthworm casts were observed to have significantly lower absolute abundances of ARGs, especially qnrS, tetM, and tetX with the removal exceeding 90%. Cell-free and –associated ARGs (except sul1 and tetG) had equivalent contributions to the attenuation of each ARG. Remarkable reductions of bacterial number and alpha diversity (chao1 and Shannon) were detected in the casts. Spearman correlation analysis between the targeted genes and bacterial community indicates that twelve different phyla mainly including Acidobacteria, Euryarchaeota, Deinococcus-Thermus, Chlorobi, Firmicutes, Fibrobacteres, and Proteobacteria are the potential ARGs hosts, suggesting that the fate and behaviour of these hosts during gut digestion of EAS by earthworms substantially determined the dynamics of the ARGs. These findings increase our understanding of earthworm gut digestion as an important process for the attenuation of ARGs in EAS, and contribute towards preventing their release into the total environment.