Improved regression model for anaerobic ammonium oxidation by repeated and prolonged batch assay under stressful salinity and pH conditions

The aim of this study was to propose repeated and prolonged batch (RPB) assay as a promising specific anammox activity (SAA) methodology assessing the anammox activity under stressed salinity and pH conditions. Response surface analysis (RSA) was used as a regression tool to evaluate statistical significance. The feasibility of RPB was investigated at 0 to 15 g-NaCl/L of salinity and pH 6 to 8 with reflecting the results of preliminary SAA. As a result, conventional SAA was statistically insignificant. In addition, the RSA results obtained from repeated batch did not meet the statistical significance despite ten times iterative reaction. Interestingly, the RPB assay (i.e., applied both repeated and prolonged reaction) was effective to obtain the reliable results. Candidadus Brocadia and Candidadus Jettenia were functional anammox microbiome during RPB. Outcomes of this study suggest that RPB assay can be applied to accurately determine the anammox activity under various stressful conditions.