Deciphering correlation between chromaticity and activity of anammox sludge

The red color is the most striking character of anaerobic ammonium-oxidizing bacteria (AnAOB) which has been used to estimate the anammox activity roughly. However, the quantitative relationship between the color and activity of anammox sludge still remains unknown. In this study, the chromaticity, activity and their correlation were systematically investigated at different steady-state nitrogen loading rates. The chromaticity of anammox sludge was digitalized by the CIE L*a*b* color space. The results revealed that the average chroma value was found to be significantly correlated with specific anammox activity (r = 0.940, p < 0.01) and the cluster centers of chromaticity coordinates (a*, b*) of anammox sludge were established to define the typical working states of anammox system. The visible spectra of anammox sludge were proved to originate from the cytochrome c. The correlation between chroma and heme c concentration of anammox sludge was consistent with the fully-reduced cytochrome c and the chroma was determined by both content and redox ratio of cytochrome c. The chromaticity of anammox sludge was able to be linked with the anammox activity via reduced cytochrome c content. The gene abundance of cytochrome c synthetase linked the chromaticity with AnAOB quantity via total cytochrome c content, while the enzyme activity of octaheme hydrazine dehydrogenase linked the chromaticity with AnAOB activity via reduced cytochrome c ratio. Moreover, the redundancy analysis proved that heme c, as the key component of cytochrome c, was the most important explanatory variable accounting for the maximum 69.6% of the total variation of the anammox community, which correlated positively with the relative abundance of dominant AnAOB (Candidatus Kuenenia). This work aimed at demonstrating the chromaticity of anammox sludge could be developed as an alternative intuitive anammox activity indicator which will promote the monitoring and optimization of anammox process.