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The inferred metagenomes analysis demonstrated that the shifts in functional microorganisms were related to variation in the main metabolic pathways. The specific activities of anammox and ASO both are regarded as key indicators for the successful start-up of bioreactor. This work revealed a novel technique for the preservation of anammox consortia and might be a potential strategy for overcoming the drawback of long start-up time. A lack of understanding exists regarding how freshwater species will respond to increases in temperature