A two-compartment microcosm was used to investigate the role of arbuscular mycorrhizal fungus (AMF) hyphae and earthworm in altering soil microbial community and OTC degradation. Treatments comprised OTC-contaminated hyphal compartments with or without AMF hyphae and with or without earthworms. Results indicated both AMF hyphae and earthworms accelerated OTC degradation; two degradation products were identified as 4-epi-oxytetracycline (EOTC) and 2-acetyl-2-decarboxamido-oxytetracycline (ADOTC). Q-PCR results indicated that both earthworms and AMF hyphae increased 16s rDNA gene, enhancing OTC degradation consequently. Illumina sequencing of the 16S rRNA genes showed that AMF hyphae and earthworm altered bacterial community. Earthworms stimulated the growth of class Anaerolineae, family Flavobacteriaceae, Genus Pseudomonas, reducing OTC residues. AMF hyphae significantly increased the abundance of family Pirellulaceae, genus Glycomyces, and Nonomuraea which had a negative correlation with EOTC, accelerating OTC degradation. When used together, AMF hyphae and earthworms enhanced OTC degradation by stimulating class Anaerolineae and family Flavobacteriaceae. (C) 2017 Elsevier B.V. All rights reserved.