Two polarity reversal modes lead to different nitrate reduction pathways in bioelectrochemical systems.

Polarity reversal is one of the effective strategies to rapidly start up denitrifying BESs，but the long-term performances of the denitrifying BESs operated under polarity reversal receive little attention. This study investigated the effects of periodic polarity reversal (PPR) and polarity reversal once only (PRO) on the long-term performances of denitrifying BESs. Repeatable oxidative and reductive currents were observed in the BESs obtained by PPR (PPR-BESs). The peak reductive currents of the PPR-BESs reached 0.95 A/m2, and nitrate was mainly removed by dissimilatory nitrate reduction to ammonium pathway with removal rates higher than 95 %. In contrast, the peak reductive currents of the BESs obtained by PRO (PRO-BESs) progressively decreased from 1.01 A/m2 to 0.12 A/m2. The nitrate removal rates of the PRO-BESs were <50 %, and the product of nitrate reduction turned to N2 instead of ammonium. 16S rDNA sequencing and metatranscriptomic analysis revealed that Geobacter capable of bidirectional extracellular electron transfer (EET) and Afipia capable of autotrophic growth were the dominant genera in the two types of BESs. Outer membrane cytochrome c and formate dehydrogenase were potentially involved in the cathodic electron uptake. These findings contribute to a better understanding of the EET mechanisms of electroautotrophic denitrifiers.