Acclimation of Microbial Consortia to Ammonia and Salt in Methane Fermentation

Abstract

As methane fermentation is inhibited by ammonia derived from organic waste, anaerobic microbial communities tolerant to enriched wastewater with high concentrations of ammonia and salt must be obtained for methane fermentation. Therefore, acclimation cultures were prepared in bottles for 60–80 weeks with artificial wastewater medium added every 2 weeks, using three types of sludge from wastewater treatment plants in food factories. These cultures were maintained without substantially decreasing methanogenesis and gradually increasing NH4-N and salt concentrations to 5 and 34 g/L, respectively, via the accumulation of ammonia and salt through anaerobic digestion and direct addition. The culture did not show the severe inhibition of methanogenesis or the accumulation of volatile fatty acids (VFAs) such as acetic and propionic acids. The analysis of bacterial consortia in the acclimated sludge based on the 16S rRNA sequence showed that hydrogenotrophic methanogenic bacteria of the genus Methanoculleus were dominant among archaea, whereas bacteria from the orders Clostridiales and Bacteroidales were dominant among eubacteria. Further, VFA-assimilating bacteria, including synthetic acetate-oxidizing bacteria coupled with hydrogenotrophic Methanoculleus to convert methane from acetate, were present to prevent the excessive accumulation of VFAs in the acclimation culture. The proposed acclimation process can enhance the anaerobic digestion of wastewater for methane production.

Keywords

• acclimation
• ammonia
• methane fermentation
• microbial consortia
• repetitive batch culture
• salt