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Researchers believe that adding conductive mediums in electrochemically active biofilms (EABs) would improve the sensitivity of EAB-based biosensor for real-time water quality early-warning through facilitating the extracellular electron transfer (EET), which has been hardly evidenced mostly because naturally formed EABs employed in previous biosensor studies were recognized distinct and incapable of delivering comparable electrical signals. By preparing artificial EABs where Shewanella oneidensis MR-1 was encapsulated in sodium alginate (SA), this study solved how polypyrrole (PPy) as conductive medium would affect the sensitivity of EAB-based biosensor, as well as mass transfer of toxicant during this process. Different mass ratios (0.125:1, 0.25:1 and 1:1) of PPy over SA were tested, and the sensitivity promoted by 20%, 15% and 6%, respectively. Results indicated that a small amount of PPy addition (PPy: SA = 0.125: 1 in mass ratio) was more effective to increase the biosensor's sensitivity compared to larger amount of PPy employed in EAB. This was when improved conductivity introduced by PPy would dominate in affecting the sensitivity over contrarily weakened mass transfer in the meantime. • Artificial electroactive biofilms containing polypyrrole were prepared. • Polypyrrole increases conductivity as well as decreases mass transfer. • Small amount of polypyrrole better increased biosensor's sensitivity. [ABSTRACT FROM AUTHOR]