Minimizing extracellular DNA improves the precision of microbial community dynamic analysis in response to thermal...

Extracellular DNA (exDNA) can induce bias when evaluating the microbiota in wastewater treatment systems, particularly when cell lysis caused by thermal hydrolysis pretreatment (THP) releasing abundant DNA. However, the influence of such exDNA is still unknown. Accordingly, this study applied a pretreatment strategy for DNA extraction with proteinase K and DNase Ⅰ to minimize the influence of exDNA when evaluating the sludge microbiota. Lactobacillus and Peptostreptococcus were confirmed as the main THP-resistant microorganisms. Gram-positive bacteria were more resistant to THP, implying that the presence of a cell wall could promote THP resistance in bacteria. Moreover, the ability to form spores did not affect the resistance of bacteria to THP. These findings showed that resistant microbiota could be effectively evaluated by excluding exDNA, which can provide important insights into the understanding of microbiota dynamic and the effects of pretreatment on the precision of microbiota analysis in sludge.