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To aid in their digestion of wood, lower termites are known to harbour a diverse community of prokaryotes as well as parabasalid and oxymonad protist symbionts. One of the best-studied lower termite gut communities is that of  Zootermopsis angusticollis  which has been known for almost 100 years to possess 3 species of  Trichonympha  ( T. campanula ,  T. collaris , and  T. sphaerica ), 1 species of  Trichomitopsis  ( T. termopsidis),  as well as smaller flagellates. We have re-assessed this community by sequencing the small subunit (SSU) rRNA gene and the internal transcribed spacer (ITS) region from a large number of single  Trichonympha  and  Trichomitopsis  cells for which morphology was also documented. Based on phylogenetic clustering and sequence divergence, we identify 3 new species:  Trichonympha postcylindrica ,  Trichomitopsis minor , and  Trichomitopsis parvus  spp. nov. Once identified by sequencing, the morphology of the isolated cells for all 3 new species was re-examined and found to be distinct from the previously described species:  Trichonympha postcylindrica  can be morphologically distinguished from the other  Trichonympha  species by an extension on its posterior end, whereas  Trichomitopsis minor  and  T. parvus  are smaller than  T. termopsidis  but similar in size to each other and cannot be distinguished based on morphology using light microscopy. Given that  Z. angusticollis  has one of the best characterized hindgut communities, the near doubling of the number of the largest and most easily identifiable symbiont species suggests that the diversity of hindgut symbionts is substantially underestimated in other termites as well. Accurate descriptions of the diversity of these microbial communities are essential for understanding hindgut ecology and disentangling the interactions among the symbionts, and molecular barcoding should be a priority for these systems.

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Single-Cell DNA Barcoding Using Sequences from the Small Subunit rRNA and Internal Transcribed Spacer Region Identifies New Species of Trichonympha and Trichomitopsis from the Hindgut of the Termite Zootermopsis angusticollis