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Perfluorooctanoic acid (PFOA) is a type of toxic per- and poly-fluoroalkyl substance (PFAS) commonly found in groundwater due to its use in firefighting and industrial applications. The main purpose of this study was to investigate the influence of PFOA shock on the biological performance of a hydrogen-driven bioreactor for nitrate and arsenate removal. Four hydrogen-driven removal reactors (HdBRs) used for the simultaneous removal of nitrate and arsenal were operated with concentrations of either 0, 1, 5, and 10 mg/L of PFOA to induce shock on the systems and examine the corresponding bacterial response. Our results showed that PFOA shock inhibited and decreased the maximum hydrogen-driven arsenate removal rate. Principal Component Analysis (PCA) confirmed that this performance decrease occurred due to a bacterial strike triggered by PFOA shock. PFOA toxicity also led to protein secretion and sludge density decreases. Bacterial analyses showed shifts in the community population due to PFOA shock. The dominant bacteria phylum Proteobacteria became more abundant, from 41.24% originally to 48.29% after exposure to 10 mg/L of PFOA. Other phyla, such as Euryarchaeota and Bacteroidetes, were more tolerant to PFOA shock. Although some of the predominant species within the sludge of each HdBR exhibited a decline, other species with similar functions persisted and assumed the functional responsibilities previously held by the dominant species. [Display omitted] • PFOA possess negative impacts on H 2 -driven biological nitrate and arsenate removal. • PFOA caused significant protein secretion and sludge particle size increases. • PFOA shock led to noticeable microbial population shifts within the HdBRs. • Members of Euryarchaeota and Bacteroidetes phyla could better tolerate PFOA shock. [ABSTRACT FROM AUTHOR]