Antibiotic and antibiotic resistance genes (ARGs) have been considered as emerging environmental contaminants and possess potential crisis to global public health. However, little is known about the differences between various configurations of two-stage combinations of constructed wetlands (CWs) on antibiotics and ARG removal from wastewater. In the study, three configurations of two-stage hybrid CWs (horizontal subsurface flow-down-flow vertical subsurface flow CWs, HF-DVF; horizontal subsurface flow-up-flow vertical subsurface flow CWs, HF-UVF; down-flow vertical subsurface flow-up-flow vertical subsurface flow CWs, DVF-UVF) were operated to evaluate their ability to remove high-concentration antibiotics (tilmicosin-TMS and doxycycline-DOC), ARGs (seven tet genes and three erm genes), intI1, 16S rRNA, and nutrients from goose wastewater. The results showed that all three hybrid CWs could remove more than 98% of TMS and DOC from wastewater, without significant difference among treatments (p > 0.05). For ARGs, DVF-UVF showed significantly higher removal efficiencies of intI1, ermB, ermC, ermF, tetW, and tetG compared to HF-UVF (p < 0.05), mainly because they might remove and arrest growth of bacteria. The relatively high removal efficiencies of NH₄⁺-N, NO₃^-N, and NO₂^--N were also observed from DVF-UVF, ranging from 87% to 95% (p > 0.05), indicating that anaerobic ammonium oxidation (anammox) might be established in the CWs. Our results demonstrate that the removal performances of antibiotics using two-stage hybrid CWs are not affected by the combined configuration, whereas the combination of DVF and UVF CWs perform better on the removal of ARGs and nutrients compared with HF-DVF and HF-UVF CWs.

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