The application of partial nitritation-anammox (PN/A) under mainstream conditions can enable substantial cost savings at wastewater treatment plants (WWTPs), but how process conditions and cell physiology affect anammox performance at psychrophilic temperatures below 15 °C remains poorly understood. We tested 14 anammox communities, including 8 from globally-installed PN/A processes, for (i) specific activity at 10-30 °C, (ii) composition of membrane lipids, and (iii) microbial community structure. We observed that membrane composition and cultivation temperature were closely related to the activity of anammox biomasses. The size of ladderane lipids and the content of bacteriohopanoids were key physiological components related to anammox performance at low temperatures. We also indicate that the adaptation of mesophilic cultures to psychrophilic regime necessitates months, but in some cases can take up to 5 years. Interestingly, biomass enriched in the marine genus "Candidatus Scalindua" displayed outstanding potential for nitrogen removal from cold streams. Collectively, our comprehensive study provides essential knowledge of cold adaptation mechanism, will enable more accurate modelling and suggests highly promising target anammox genera for inoculation and set-up of anammox reactors, in particular for mainstream WWTPs.

Delft University of Technology Department of Biotechnology Building 58 Van der Maasweg 9 2629 HZ Delft

Radboud University Department of Microbiology Institute for Water and Wetland Research 1Heyendaalseweg 135 6525 ED AJ Nijmegen The Netherlands

University of Chemistry and Technology Prague Department of Biochemistry and Microbiology Technická 5 166 28 Prague Czechia

University of Chemistry and Technology Prague Department of Food Analysis and Nutrition Technická 5 166 28 Prague Czechia

University of Chemistry and Technology Prague Department of Water Technology and Environmental Engineering Technická 5 166 28 Prague Czechia

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Effect of temperature on the compositions of ladderane lipids in globally surveyed anammox populations