Industrial-scale aerobic composting of livestock manures with the addition of biochar: Variation of bacterial community and antibiotic resistance genes caused by various composting stages.

The presence of large amounts of antibiotic resistance genes (ARGs) in livestock manures poses an impending, tough safety risk to ecosystems. To investigate more comprehensively the mechanisms of ARGs removal from industrial-scale composting of livestock manure based on biochar addition, we tracked the dynamics of bacterial community and ARGs at various stages of aerobic composting of livestock manures with 10% biochar. There were no significant effects of biochar on the bacterial community and the profiles of ARGs. During aerobic composting, the relative abundance of ARGs and mobile genetic elements (MGEs) showed overall trends of decreasing and then increasing. The key factor driving the dynamics of ARGs was bacterial community composition, and the potential hosts of ARGs were Caldicoprobacter, Tepidimicrobium, Ignatzschineria, Pseudogracilibacillus, Actinomadura, Flavobacterium and Planifilum. The retention of the thermophilic bacteria and the repopulation of the initial bacteria were the dominant reasons for the increase in ARGs at maturation stage. Additionally, among the MGEs, the relative abundance of transposon gene was substantially removed, while the integron genes remained at high relative abundance. Our results highlighted that the suitability of biochar addition to industrial-scale aerobic composting needs to be further explored and that effective measures are needed to prevent the increase of ARGs content on maturation stage.

Variations in antibiotic resistance genes and removal mechanisms induced by C/N ratio of substrate during composting.

For a comprehensive insight into the potential mechanism of the removal of antibiotic resistance genes (ARGs) removal induced by initial substrates during composting, we tracked the dynamics of physicochemical properties, bacterial community composition, fungal community composition, the relative abundance of ARGs and mobile genetic genes (MGEs) during reed straw and cow manure composting with different carbon to nitrogen ratio. The results showed that the successive bacterial communities were mainly characterized by the dynamic balance between Firmicutes and Actinobacteria, while the fungal communities were composed of Ascomycota. During composting, the interactions between bacteria and fungi were mainly negative. After composting, the removal efficiency of ARGs in compost treatment with C/N ≈ 26 (LL) was higher than that in compost treatment with C/N ≈ 35 (HL), while MGEs were completely degraded in HL and enriched by 2.3% in LL. The large reduction in the relative abundance of ARGs was possibly due to a decrease in the potential host bacterial genera, such as Advenella, Tepidimicrobium, Proteiniphilum, Acinetobacter, Pseudomonas, Flavobacteria and Arcbacter. Partial least-squares path modeling (PLS-PM) revealed that the succession of bacterial communities played a more important role than MGEs in ARGs removal, while indirect factors of the fungal communities altered the profile of ARGs by affecting the bacterial communities. Both direct and indirect factors were affected by composting treatments. This study provides insights into the role of fungal communities in affecting ARGs and highlights the role of different composting treatments with different carbon to nitrogen ration on the underlying mechanism of ARGs removal.