Diversity of anaerobic fungi within cow manure determined by ITS1 analysis

The diversity of anaerobic fungi was evaluated in cow semiliquid manure obtained from input homogenizing tank of biogas plant. Among three sets of tested primers, the combination of fungal universal ITS1F and Neocallimastigales specific Neo QPCR Rev primers was selected and used for the construction of clone library. Eighty-four new complete internal transcribed spacers (ITS1) and partial 5.8S rDNA sequences generated within this study were analyzed by Bayesian inference and assigned to an existing order of the Neocallimastigales. Sixty-seven % of sequences were affiliated with Cyllamyces, 24 % with Piromyces, 7 % with Anaeromyces, only 2 % with Neocallimastix, and no sequences with Orpinomyces. According to Bayesian analysis the genus Caecomyces was polyphyletic and disappeared from the presented ITS-based phylogram. This study gave a first insight into the diversity of anaerobic fungi in cow manure, where the prevalence of fungi with bulbous morphology was indicated.