Mizuho Muramatsu

Molecular Phylogenetic Diversity of the Bacterial Community in the Gut of the Termite Coptotermes formosanus

The phylogenetic diversity of the bacterial community in the gut of the termite Coptotermes formosanus was investigated using a 16S rRNA gene clone library constructed by PCR. After screening by restriction fragment length polymorphism (RFLP) analysis, 49 out of 261 clones with unique RFLP patterns were sequenced and phylogenetically analyzed. Many of the clones (94%) were derived from Bacteroidales, Spirochaetes, and low G+C content gram-positive bacteria consisting of Clostridiales, Mycoplasmatales, Bacillales, and Lactobacillales. In addition, a few clones derived from Actinobacteria, Proteobacteria, Planctomycetes, Verrucomicrobia, and the candidate phylum “Synergistes” were also found. The most frequently identified RFLP type, BCf1-03, was assigned to the order Bacteroideales, and it constituted about 70% of the analyzed clones. The phylogenetic analysis revealed that the representative clones found in this study tended to form some clusters with the sequences cloned from the termite gut in several other studies, suggesting the existence of termite-specific bacterial lineages.

Phylogenetic Analysis of the Gut Bacterial Microflora of the Fungus-Growing Termite Odontotermes formosanus

We constructed a bacterial 16S rRNA gene clone library from the gut microbial community of O. formosanus and phylogenetically analyzed it in order to contribute to the evolutional study of digestive symbiosis and method development for termite control. After screening by restriction fragment length polymorphism (RFLP) analysis, 56 out of 280 clones with unique RFLP patterns were sequenced and phylogenetically analyzed. The representative phylotypes were affiliated to four phylogenetic groups, Firmicutes, the Bacteroidetes/Chlorobi group, Proteobacteria, and Actinobacteria of the domain Bacteira. No one clone affiliated with the phylum Spirochaetes was identified, in contrast to the case of wood-feeding termites. The phylogenetic analysis revealed that nearly half of the representative clones (25 phylotypes) formed monophyletic clusters with clones obtained from other termite species, especially with the sequences retrieved from fungus-growing termites. These results indicate that the presence of termite-specific bacterial lineages implies a coevolutional relationship of gut microbes and host termites.

Armatimonas rosea gen. nov., sp. nov., of a novel bacterial phylum, Armatimonadetes phyl. nov., formally called the candidate phylum OP10

A novel aerobic, chemoheterotrophic bacterium, strain YO-36(T), isolated from the rhizoplane of an aquatic plant (a reed, Phragmites australis) inhabiting a freshwater lake in Japan, was morphologically, physiologically and phylogenetically characterized. Strain YO-36(T) was Gram-negative and ovoid to rod-shaped, and formed pinkish hard colonies on agar plates. Strain YO-36(T) grew at 20-40 °C with optimum growth at 30-35 °C, whilst no growth was observed at 15 °C or 45 °C. The pH range for growth was 5.5-8.5 with an optimum at pH 6.5. Strain YO-36(T) utilized a limited range of substrates, such as sucrose, gentiobiose, pectin, gellan gum and xanthan gum. The strain contained C(16 : 0), C(16 : 1), C(14 : 0) and C(15 : 0) as the major cellular fatty acids and menaquinone-12 as the respiratory quinone. The G+C content of the genomic DNA was 62.4 mol%. Phylogenetic analysis based on 16S rRNA gene sequences revealed that strain YO-36(T) belonged to the candidate phylum OP10 comprised solely of environmental 16S rRNA gene clone sequences except for two strains, P488 and T49 isolated from geothermal soil in New Zealand; strain YO-36(T) showed less than 80 % sequence similarity to strains P488 and T47. Based on the phylogetic and phenotypic findings, a new genus and species, Armatimonas rosea gen. nov., sp. nov., is proposed for the isolate (type strain YO-36(T)  = NBRC 105658(T)  = DSM 23562(T)). In addition, a new bacterial phylum named Armatimonadetes phyl. nov. is proposed for the candidate phylum OP10 represented by A. rosea gen. nov., sp. nov. and Armatimonadaceae fam. nov., Armatimonadales ord. nov., and Armatimonadia classis nov.

Fatty acids of astaxanthin esters in krill determined by mild mass spectrometry

Krill is a major source of astaxanthin, which has strong antioxidant activity. Fractions with astaxanthin monoesters and diesters of Antarctic krill Euphausia superba were isolated. Astaxanthin esters were separated by C18-HPLC depending on the number of carbons and double bonds of esterified fatty acid(s). Small amounts of other lipids remained in the samples, but relative molecular masses of carotenoid esters could be measured by field desorption mass spectrometry without fragmentation and interference from contaminant lipids. The fatty acids were determined by calculation of difference between astaxanthin and astaxanthin esters. Only five kinds of fatty acids, dodecanoate, tetradecanoate, hexadecanoate, hexadecenoate and octadecenoate, were detected. Fast atom bombardment mass spectrometry and secondary ion mass spectrometry showed similar spectra. The fatty acid composition in astaxanthin esters was different from those in krill lipids. Therefore, determination of fatty acids in carotenoid esters by a combination of HPLC elution profile and mild mass spectrometry is found to be a useful tool.

Termite-Regulated Fungal Monoculture in Fungus Combs of a Macrotermitine Termite Odontotermes formosanus

Abstract The mechanism of the exclusive growth of Termitomyces in fungus combs with fungi-growing termites, O. formosanus was examined using laboratory scale fungus combs. In the combs without the termites, vigorous growth of unidentified fungi was observed although no significant change was found in the case of the combs with termites. In addition, these results were reproducible even when incubated in a separated dish, suggesting that the physicochemical conditions were not the reason for the growth. With the molecular based analysis for the microbial communities in the combs, monoculture of the Termitomyces in the combs with termites was confirmed while the bacterial communities were independent either with or without termites. Possible mechanism of the exclusive growth of Termitomyces, such as the selective grazing of pathogenic fungi or contribution of antifungal activity giving actinomycetes were also discussed.

Shinella yambaruensis sp. nov., a 3-methyl-sulfolane-assimilating bacterium isolated from soil.

A bacterial strain, designated MS4(T), was isolated from soil in the Ryukyu Archipelago, Japan. The bacterium grew with 3-methyl sulfolane as sole sulfur source. Phylogenetic analysis based on 16S rRNA gene sequences suggested that strain MS4(T) belonged to the genus Shinella; it was closely related to the type strains of Shinella granuli and Shinella zoogloeoides (16S rRNA gene sequence similarities of 98.2 and 96.7 %, respectively). Strain MS4(T) was a Gram-negative, non-motile, rod-shaped, aerobic bacterium. The major respiratory quinone was ubiquinone-10 and the predominant cellular fatty acid was C(18 : 1)omega7c. The DNA G+C content was 66.4 mol%. Based on phylogenetic and phenotypic traits, it was concluded that the organism represents a novel species in the genus Shinella for which the name Shinella yambaruensis sp. nov. is proposed; the type strain is MS4(T) (=NBRC 102122(T)=DSM 18801(T)).