從微生物菌相生態實驗中，於白蟻腸道及菌圃中發現變形菌門、厚壁菌門等不同的細菌菌株，及些許無法培養的菌株，其中主要的細菌菌株屬厚壁菌門。而實驗所分離到的真菌主要是雞肉絲菇與一些小團體，如酵母菌。接著將這些分離純化到的細菌菌株與侵入性的microfungi Trichoderma harzianum進一步研究其相互作用關係。藉由這些研究得知腸道共生菌在調控菌圃侵入性microfungi過渡生長上扮演著重要的角色。實驗發現芽孢桿菌會抑制T. harzianum的生長因而促進雞肉絲菇的生長。因此，它被視為啟動的真菌生長的互惠者。隨後在分離純化到的菌株中分析其木質纖維素降解、固氮及生物制氫的功能。
The black winged subterranean termite Odontotermes formosanus is the only known macrotermitine termite in Taiwan. These termites construct large colonies with numerous fungus combs in subterranean nest and have great impact on the degradation of plant litter. The fecal material of O. formosanus containing the ingested plant material is made into a small ventilated structure, the fungus comb or the “fungus garden” to which the mycelium of the symbiotic fungi Termitomyces grows. Previous studies portray the symbiotic relationship between Odontotermes formosanus and Termitomyces species. By this relationship, the termite acquires nitrogenous compounds from the fungal hyphae of Termitomyces and in turn these fungi are provided with a suitable habitat for their growth and sustenance.
So far, the fruiting body of Termitomyces has not been cultivated in the laboratory in the absence of termites. This research has gained significance as gut symbionts are used to enhance the growth of Termitomyces mycelia by culturing. Therefore it is necessary to study the functioning of the O. formosanus termite-gut fungus comb symbionts under in vitro conditions. For this research, the microbial communities harboring in the gut and fungus comb of this termite are analyzed by both culture dependent and culture independent methods to better understand the ecology and community structure of their microflora. For the detection of bacterial and fungal communities by culture independent methods, denaturing gradient gel electrophoresis (DGGE) and clonal selection are performed; and for culture dependent methods, microbial communities are isolated using different culture mediums. These microbes are hypothesized to contribute to cellulose-hemicellulose hydrolysis, gut fermentation, nutrient production, fungus comb ageing and the initiation of the growth of the symbiotic fungus Termitomyces.
From the microbial ecological studies by culture dependent and independent methods in the termite gut and the fungus comb, the different bacterial strains are identified to belong to Proteobacteria, Firmicutes and other uncultured bacteria, of which Firmicutes are predominant. The dominant fungal isolate belong to Termitomyces species where as other minor groups like yeasts are also detected. The bacterial strains isolated by the cultural approach are further used for the interaction studies along with the invasive microfungi Trichoderma harzianum. These studies are performed to know if the gut symbionts contributed an important role in regulating the growth of other invasive microfungi overgrowing on the fungus comb. The Bacillus strains are detected to suppress the growth of T. harzianum and promote the growth of the dominant fungi Termitomyces. Therefore, they are considered as mutualists that initiate the growth of the fungi. The functional roles of the isolated strains are assessed for lignocellulosic degradation, nitrogen fixation and biohydrogen production.
In addition, the gut symbionts from the macrotermitine termite Odontotermes formosanus, the cellulolytic Bacillus and fermentative Clostridium are studied in batch experiments using different carbon substrates to biomimic the termite gut for hydrogen production. Their fungus comb ageing and the in vitro lignocellulosic degradation of the mango tree substrates by the synergistic interaction of Bacillus, Clostridium and Termitomyces are detected by Solid-state NMR. From the results, Bacillus species act as mutualists, by initiating an anaerobic environment for the growth of Clostridium for biohydrogen production and enhancing the lignocellulosic degradation of substrates in the presence of Termitomyces in vitro and in vivo. Thus, the synergistic collaboration of these three microbes can be used for termite-derived biofuel process technology. The chemical composition of the fungus comb is monitored and the degradation pattern of the organic components as a result of ageing is similar to the mango tree substrates that are incubated with cocultures of Bacillus, Clostridium and Termitomyces. The overall research is focused on studying the microbial community in the termite gut and fungus comb system of O. formosanus, their functional aspects and biomimicking their microbial process for biohydrogen production.