Genetic profiling via a novel PCR-RFLP method enabled identification of four genera of anaerobic gut fungi isolated from nyala, giraffe, and zebra hosts.

LSU-based PCR-RFLP using AluI and Hyp188I digestion identified distinct anaerobic gut fungal genera, enabling isolation of Piromyces (NY08), Neocallimastix cameroonii, Feramyces austinii, and Khoyollomyces ramosus from zoo herbivores.

• Why it matters: Axenic culture of anaerobic gut fungi is resource-intensive, limiting functional studies and niche mapping within Neocallimastigomycota in herbivore gut microbiomes.
• What they did: The team combined morphological and functional screening with an in silico–guided PCR-RFLP strategy, then validated genus calls by confirmatory ITS or LSU sequencing and amplicon sequencing of host communities.
• The result: Amplicon sequencing showed these species dominated AGF communities in their respective hosts, and enrichment conditions were linked to cultivability, supporting a workflow for efficient AGF isolation for biotechnology.

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