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200539 https://hdl.handle.net/2286/R.2.N.200539 http://rightsstatements.org/vocab/InC/1.0/ http://creativecommons.org/licenses/by-nc-sa/4.0 2025-05 32 pages Willis, Maya Gile, Gillian Hileman, Jonathon Barrett, The Honors College School of Life Sciences School of Applied Sciences and Arts Coptotermes formosanus, an invasive termite species from East Asia, causes around $2 billion in damage and infestation control annually, and is a significant risk to the United States economy and infrastructure. C. formosanus is known to harbor 5 different species of eukaryotic protists in its hindgut: Pseudotrichonympha grassii, Holomastigotoides hartmanni, Holomastigotoides minor, Cononympha leidyi, and Cononympha koidzumii. Each of these protists has an additional symbiotic relationship with multiple prokaryotic endo- and/or ectosymbionts. Ectosymbionts associated with Holomastigotoides – such as spirochetes and Parendomicrobium – are likely able to utilize H2, ferment glucose and hexuronate and metabolize monoethanolamine, but their complete diversity and function is unknown. The goal of this study was to elucidate the localization and function of a new prokaryotic ectosymbiont, from the order Bacteroidales, associated with Holomastigotoides using fluorescent in-situ hybridization (FISH) and metagenome-assembled genome (MAG) analysis. The symbiont lies close and flat to the cell membrane and is not localized to a specific end of the cell or associated with the flagellar lines. The bacterial cells vary in size, between 1 micron and 10 microns, along with their population size. According to MAG analysis, the symbiont is potentially capable of: D-galacturonate biosynthesis, cysteine and arginine biosynthesis, PRPP biosynthesis, beta-oxidation/acyl-CoA and PE biosynthesis, and many other possible functions. An understanding of how prokaryotic ectosymbionts contribute to the overall metabolism for their host protist and termite illuminates the ecological importance of this mutual symbiosis. Microbial Eukaryotic Diversity Prokaryotic Diversity Termite Symbiosis Evolution Phylogenetics Bioinformatics Fluorescent in-situ Hybridization and Metagenomic Analysis of a New Bacteroidales Symbiont of Holomastigotoides within the Coptotermes formosanus Hindgut