Effects of coexistence of tetracycline, copper and microplastics on the fate of antibiotic resistance genes in manured soil

Highlights: • Environmental microplastics (EM) increased the bioavailable Cu and TC in soil. • EM magnified co-selection of TC and Cu for antibiotic resistance in manured soil. • Intl1 played a great role in ARG propagation in soil environments and plastisphere. • EM could be the hotspot of antibiotic-resistant bacteria and intl1 gene in soil. • EM in manured soil could increase the risk of ARGs pollution from animal manure. The coexistence of antibiotics, heavy metals and microplastics is becoming commonplace and may affect antibiotic resistance in manured soil. The current understanding of the role of microplastics in soil with combined pollution of antibiotics, heavy metals and antibiotic resistance genes (ARGs) is limited. Here, the effects of the coexistence of tetracycline (TC), Cu and environmental microplastics (EM) on the fate of nine ARGs and three heavy metal resistance genes in agricultural soil were investigated by batch and microcosm experiments. EM were obtained by exposing virgin microplastics to soil environments for 80 days, which exhibited higher adsorption affinity for Cu and TC than soil particles and virgin microplastics. 1% EM in soil increased bioavailable concentrations of TC and Cu by 79–138% and 88–135%, respectively, and decreased TC dissipation from 11.79 mg kg⁻¹ to 3.08 mg kg⁻¹. Correspondingly, the total relative abundances of target ARGs increased by 219–348%. The significant correlations of tetG, tetB, tetQ, sul2, sul1 and intl1 with bioavailable fractions of TC and Cu in soil environments were revealed by network analysis. Moreover, scanning electron micrographs showed the special plastisphere around EM. Attributed to the biofilm generation and higher pollutant accumulation in the plastisphere, EM could be the source of antibiotic-resistant bacteria and ARGs in soil environments. Structure equation models further identified that indirect effects of EM acted a major role in the propagation of ARGs by altering soil properties, soil microbial diversity and intl1 abundance. This study revealed that EM could increase the stimulative effects of Cu and TC on antibiotic resistance and magnify the environmental risk of manure application in soil environments.