Effects of passivators on antibiotic resistance genes and related mechanisms during composting of copper-enriched pig manure

Due to the intensive use of feed additives in livestock farming, animal manure has become a hotspot for antibiotics, heavy metals, and antibiotic resistance genes (ARGs). Unlike antibiotics, heavy metals cannot be degraded during composting and thus could pose a persistent co-selective pressure in the proliferation of antibiotic resistance. Passivators are commonly applied to immobilize metals and improve the safety of compost. However, little is known about the effects of various passivators on ARGs and mobile genetic elements (MGEs) during composting and the underlying mechanisms involved. Thus, three typical passivators (biochar, fly ash, and zeolite) were applied during the composting of copper-enriched pig manure, and their effects on ARGs, copper resistance genes, MGEs, and the bacterial communities were examined. Compared to the control, all passivator treatments reduced the abundances of at least six ARGs (tetC, tetG, tetQ, tetX, sul1, and ermB) by 0.23–1.09 logs and of two MGEs (intI1 and ISCR1) by 26–85% after composting. Biochar and fly ash also significantly reduced the abundances of intI2 and Tn914/1545. In contrast, abundances of copper resistance genes were not reduced by passivators, implying that the decreased co-selective pressure may not be a major contributor to ARG reductions in this study. Procrustes analysis and redundancy analysis demonstrated that shifts in the bacterial community determined the changes in the abundances of ARGs, and the variation in MGEs and DTPA-Cu can also partially explain the ARG variance. Overall, all of three passivators can be used to reduce the health risks associated with ARGs in livestock manure, and biochar performed the best at reducing ARGs and MGEs.