Biochar induced inhibitory effects on intracellular and extracellular antibiotic resistance genes in anaerobic digestion of swine manure.

Distribution of intracellular (iARGs) and extracellular ARGs (eARGs) in manure anaerobic digestion (AD) process coupled with two types of biochar (BC and BP) were investigated. And the effects of biochar on the conjugation transfer of ARGs were explored by deciphering the interaction of biochar with bacterial stress responses, physiological metabolism and antibiotic resistances. Results showed that AD process could effectively remove all the detected eARGs with efficiency of 47.4-98.2%. The modified biochar (BP) with larger specific surface area (SSA) was propitious to decrease the absolute copy number of extracellular resistance genes. AD process could effectively remove iARGs by inhibiting the growth of host bacteria. The results of structural equation models (SEM) indicated that biochar put indirect influences on the fate of ARGs (λ = -0.23, P > 0.05). Analysis on oxidative stress levels, antioxidant capacity, DNA damage-induced response (SOS) response and energy generation process demonstrated that biochar induced the oxidative stress response of microorganisms and enhanced the antioxidant capacity of bacteria. The elevated antioxidant capacity negatively affected SOS response, amplified cell membrane damage and further weakened the energy generation process, resulted in the inhibition of horizontal transfer of ARGs.