Isolation and identification of cellulolytic anaerobic fungi and their associated methanogens from Holstein Cow

Abstract:

Abstract: ［Objective］ We studied the microbial interaction between anaerobic fungi and methanogens in the rumen of Holstein Cow.［Methods］ Co-cultures of anaerobic fungi with indigenously associated methanogen were isolated by Hungate roll-tube technique. The anaerobic fungi were identified by morphology and 4’，6 diamidino-2-phylindole nucleus staining and the methanogens were identified by 16S rＲNA gene sequencing． ［Ｒesults］A total of 28 co-cultures of anaerobic fungus with indigenously associated methanogen were obtained． The anaerobic fungi in the co-cultures were identified as monocentric genera Piromyces，Neocallimastix and Caeomyces. The indigenously associated methanogens were Methanobrevibacter olleyae like and Methanobrevibacter thaueri like strains．Four different phylotypes of fungus-methanogen co-cultures were obtained， which were Piromyces/Methanobrevibacter olleyae like strains，Neocallimastix/Methanobrevibacter olleyae like strains， Neocallimastix /Methanobrevibacter thaueri like strains and Caecomyces/Methanobrevibacter olleyae like strains．［Conclusion］Our study isolated and identified 28 co-cultures of anaerobic fungus and associated methanogens，which provided new materials for further study the mechanism of methane emission in the rumen．