Project Number: 6435-32000-010-04
Project Type: Specific Cooperative Agreement

Start Date: Sep 30, 2002
End Date: Sep 30, 2007

Objective:
The objectives of this cooperative research are to isolate, characterize and initiate axenic cultures of the symbiotic gut protozoans from the Formosan subterranean termite. Pseudotrichonympha grassei Koidzumi, Spirotrichonympha leidy Koidzumi, and Holomastigoides hartzmanii Koidzumi will be isolated from whole termites and/or isolated gut tracts from mid- to late-instar larvae of Coptotermes formosanus Shiraki. We will determine physical and biological methods to separate the species of the protozoans to be used for inocula for axenic culture or for biochemical characterization. Several known protozoan culture media and modifications of these base media will be investigated for the initiation and maintenance of one or more of the gut symbionts in culture. Various treatments of the cultured protozoans will be evaluated as possible mechanisms to control subterranean termite cellulose digestion.

Approach:
Mid- to late-instar larvae of the Formosan subterranean termite, Coptotermes formosanus Shiraki will be collected from bucket traps established in south Louisiana, particularly New Orleans and the surrounding area. Guts from freshly collected termites will be dissected into sterile media for isolation of gut protozoa and initiation of cultures. Groups of whole termites will be ground and physical methods, such as density gradient centrifugation will be utilized to separate and isolate individual species for biochemical characterization and as inoculum for axenic culture. Various published protozoan culture media will be initially utilized to attempt initiation of axenic cultures for these yet-to-be cultured organisms. Established protozoan culture media will be modified to reflect chemical composition of the termite gut. Differing culture conditions to mimic the termite gut will be provided to initiate or optimize the culture conditions for one or more of the gut symbionts.