Synergistic enhancement effect of straw-earthworms in the reduction of sulfamethoxazole and antibiotic resistance genes

Soil antibiotic pollution is a global concern. It has been confirmed that straw or earthworm can enhance microbial degradation of antibiotics in soil. However, in the C/N transformation processes of soil ecosystems, straw and earthworms are closely interconnected. Whether their interaction can further enhance microbial degradation of antibiotic pollution and the underlying mechanisms remain to be explored. This study conducted a 90 days co-incubation experiment with four treatments: straw + earthworms + sulfamethoxazole (RS-EW-SMX), straw + SMX (RS-SMX), earthworms + SMX (EW-SMX), and SMX alone (SMX). Residual SMX, its degradation intermediates, and microbial communities were monitored at multiple timepoints. Results indicated an exponential decline in SMX degradation rates across treatments. By day 90, SMX was nearly completely degraded in all treatment groups. However, the combined effect of straw and earthworms significantly enhanced the degradation efficiency of SMX. During the rapid degradation phase, SMX in above four treatments decreased from 20.0 mg kg-1 to 0.93, 1.88, 5.26 and 7.02 mg kg-1, respectively at day 10. Furthermore, the RS-EW-SMX treatment promoted SMX transformation into low-molecular-weight intermediates and increased the relative abundance of SMX-degrading bacteria by 1.35, 2.01, and 2.17-fold compared to RS-SMX, EW-SMX, and SMX, respectively. SMX degradation efficiency exhibited a strong positive linear correlation with the relative abundance of degrading bacteria across all treatments (R2 = 0.961). Concurrently, analysis revealed that straw presence facilitated the targeted enrichment of SMX-degrading bacteria within the earthworm gut, concomitant with a reduction in associated antibiotic resistance genes (ARGs). This synergistic interaction between straw and earthworms, mediated through the gut microbiome and carbon utilization, constitutes a primary mechanism underpinning the accelerated SMX degradation observed. These findings reveal a novel macrofauna-plant residues interaction mechanism for improved in situ antibiotic bioremediation, providing practical solutions for soil pollution mitigation.