The Tipula abdominalis larval hindgut microbial community presumably facilitates digestion of the lignocellulosic diet. The microbial community was investigated through characterization of bacterial isolates and analysis of 16S rRNA gene clone libraries. This initial study revealed novel bacteria and provides a framework for future studies of this symbiosis.Insects are the largest taxonomic group of animals on earth. Although a few thorough studies have shown that insects host an environment with high microbial diversity (3-5, 9, 11, 17, 24), less than 1% of described insect species have been examined for microorganisms (10). Tipula abdominalis is an aquatic crane fly ubiquitous in aquatic riparian environments. T. abdominalis larvae are shredders, a functional feeding group of insects that consume coarse particulate organic matter, primarily leaf litter. In small riparian stream ecosystems, leaf litter comprises the majority of carbon and energy inputs (26); however, many organisms are unable to degrade this lignocellulosic material, which is difficult to digest due to highly structured plant polysaccharide polymers (cellulose, hemicellulose, and lignin) and which has low nutritional value due to a high C/N ratio (15). By converting lignocellulose into a form that other organisms can use, T. abdominalis larvae influence the bioavailability of carbon and energy within the ecosystem.The larva itself is not capable of tissue-level synthesis of cellulolytic enzymes (22), and it was proposed that the larvae benefit nutritionally from microbially mediated digestion of leaf lignocellulose, providing simple fermentation products which can be used by the larvae (13). Scanning electron microscopy studies demonstrated a dense and morphologically diverse microbial community in the hindgut of T. abdominalis larvae (12). This microbial community was investigated for phylogenetic diversity and enzymatic activity towards model plant polymer substrates.Larva collection and dissection. Larvae were collected from 2nd-order streams (26) in Michigan. Hindguts were extracted and transferred to a reduced buffered salt solution (BSS) (14).