Kuroki Hiroko, Miyamoto Hiroshi, Fukuda Kazumasa, Iihara Hirotoshi, Kawamura Yoshiaki, Ogawa Midori, Wang Yan, Ezaki Takayuki, Taniguchi Hatsumi
Department of Microbiology, School of Medicine, University of Occupational and Environmental Health, Fukuoka 807-8555, Japan.
Syst Appl Microbiol. 2007 Jun;30(4):273-9. doi: 10.1016/j.syapm.2006.11.005. Epub 2007 Jan 2.
In this study, we tried to isolate legionellae from nine Legionella DNA-positive soil samples collected from four different sites contaminated with industrial wastes in Japan. Using culture methods with or without Acanthamoeba culbertsoni, a total of 22 isolates of legionellae were obtained from five of the nine samples. Identification of species and/or serogroups (SGs), performed by DNA-DNA hybridization and agglutination tests, revealed that the 22 isolates consisted of ten isolates of Legionella pneumophila including five SGs, five Legionella feeleii, and one each of Legionella dumoffii, Legionella longbeachae, and Legionella jamestownensis. The species of the remaining four isolates (strains OA1-1, -2, -3, and -4) could not be determined, suggesting that these isolates may belong to new species. The 16S rDNA sequences (1476-1488bp) of the isolates had similarities of less than 95.0% compared to other Legionella species. A phylogenetic tree created by analysis of the 16S rRNA (1270bp) genes demonstrated that the isolates formed distinct clusters within the genus Legionella. Quantitative DNA-DNA hybridization tests on the OA1 strains indicated that OA1-1 should be categorized as a new taxon, whereas OA1-2, -3, and -4 were also genetically independent in another taxon. Based on the evaluated phenotypic and phylogenetic characteristics, it is proposed that one of these isolates from the soils, OA1-1, be classified as a novel species, Legionella impletisoli sp. nov.; the type strain is strain OA1-1(T) (=JCM 13919(T)=DSMZ 18493(T)). The remaining three isolates belong to another novel Legionella species, Legionella yabuuchiae sp. nov.; the type strain is strain OA1-2(T) (=JCM 14148(T)=DSMZ 18492(T)). This is the first report on the isolation of legionellae from soils contaminated with industrial wastes.
在本研究中，我们试图从日本四个受工业废弃物污染的不同地点采集的9份军团菌DNA阳性土壤样本中分离军团菌。使用含有或不含有culbertsoni棘阿米巴的培养方法，从9份样本中的5份共获得了22株军团菌分离株。通过DNA-DNA杂交和凝集试验进行的菌种和/或血清群（SGs）鉴定显示，这22株分离株包括10株嗜肺军团菌，其中包含5个血清群，5株费氏军团菌，以及各1株杜莫夫军团菌、长滩军团菌和詹姆斯敦军团菌。其余4株分离株（菌株OA1-1、-2、-3和-4）的菌种无法确定，表明这些分离株可能属于新物种。与其他军团菌属物种相比，这些分离株的16S rDNA序列（1476 - 1488bp）相似度低于95.0%。通过对16S rRNA（1270bp）基因分析构建的系统发育树表明，这些分离株在军团菌属内形成了不同的簇。对OA1菌株进行的定量DNA-DNA杂交试验表明，OA1-1应归类为一个新分类单元，而OA1-2、-3和-4在另一个分类单元中在遗传上也是独立的。基于评估的表型和系统发育特征，建议将这些土壤分离株中的一株OA1-1分类为一个新物种，即充实土壤军团菌（Legionella impletisoli sp. nov.）；模式菌株为菌株OA1-1(T)（=JCM 13919(T)=DSMZ 18493(T)）。其余3株分离株属于另一个新的军团菌物种，即矢吹军团菌（Legionella yabuuchiae sp. nov.）；模式菌株为菌株OA1-2(T)（=JCM 14148(T)=DSMZ 18492(T)）。这是关于从受工业废弃物污染的土壤中分离军团菌的首次报道。
