Monitoring microbial communities’ dynamics during the start-up of microbial fuel cells by high-throughput screening techniques

Microbial Electrochemical Technologies are based on the use of electrochemically active microorganisms that can carry out extracellular electron transfer to an electrode while they are oxidizing the organic compounds. The dynamics and changes of the bacterial community in the anode biofilm and planktonic broth of an acetate fed batch single chamber air cathode MFC have been studied by combing flow-cytometry and Illumina sequencing techniques. At the beginning of the test, from 0 h to 70 h, microbial planktonic communities changed from four groups to two groups, as revealed by DNA content, and from three groups to one group based on the cell membrane polarization revealed by a DiOC6(3) probe. Between 4th day and 13th day, microbial communities changed from one group to a maximum of three groups, monitoring DNA content, and from one group to two based on the cell membrane polarization. The 16S rDNA gene profiling confirmed the shift in microbial communities, with Acinetobacter (39.34%), Azospirillum (27.66%), Arcobacter (4.17%) and Comamonas (2.62%) being the most abundant genera at the beginning of MFC activation. After 70 h the main genera detected were Azospirillum (46.42%), Acinetobacter (34.66%), Enterococcus (2.32%), Dysgonomonas (2.14%). Data obtained have shown that flow cytometry and illumina sequencing are useful tools to monitor “online” the changes in microbial communities during the MFCs start-up and the increase of Azospirillum and Acinetobacter genera is in good agreement with the MFC voltage generation. Moreover, monitoring planktonic populations, instead of the less accessible anode biofilm, was in good agreement with the evolution of MFC voltage. Keywords: Microbial electrochemical technologies, Flow cytometry, Bacterial community composition, Illumina high-throughput sequencing