Kutsuna Ryo, Miyoshi-Akiyama Tohru, Mori Koji, Hayashi Masahiro, Tomida Junko, Morita Yuji, Tanaka Kaori, Kawamura Yoshiaki
Laboratory of Microbiology, Aichi Gakuin University Graduate School of Pharmaceutical Science, Nagoya, Aichi, Japan.
Pathogenic Microbe Laboratory, Research Institute, National Center for Global Health and Medicine, Shinjuku, Tokyo, Japan.
Microbiol Immunol. 2019 Jan;63(1):1-10. doi: 10.1111/1348-0421.12663.
Taxonomic studies of strain PAGU 1678 , an obligately anaerobic, gram-positive, spore-forming bacterium isolated from biobreeding rat feces, were performed. This strain has been demonstrated to have the ability to exacerbate pathosis in a mouse model of dextran sulfate sodium-induced ulcerative colitis. Phylogenetic analysis based on the 16S rRNA gene showed high homology with Paraclostridium bifermentans. To clarify the correct taxonomic position of strain PAGU 1678 , a comparative taxonomic study using P. bifermentans PAGU 2008 (═JCM 1386 ) and the closely related bacterial species P. benzoelyticum PAGU 2068 (═LMG 28745 ) was carried out. Despite the close similarity of 16S rRNA gene sequences, DNA-DNA hybridization between strain PAGU 1678 and P. bifermentans PAGU 2008 was 60.03% on average, average nucleotide identity was 96.17%, and it was shown to have different genomic sequences. Biochemically, strain PAGU 1678 could be differentiated from P. bifermentans PAGU 2008 by H S production. Furthermore, strain PAGU 1678 was characterized by the presence of two phospholipids with different polarity on polar lipid analysis. In addition, strain PAGU 1678 differed from P. bifermentans PAGU 2008 in findings on whole-cell protein analysis and matrix-assisted laser desorption/ionization-time of flight mass spectrometry. On the basis of these biochemical and genetic characteristics, a novel subspecies of P. bifermentans with the name Paraclostridium bifermentans subsp. muricolitidis subsp. nov. is here proposed, with PAGU 1678 (═CCUG 72489 ═NBRC 113386 ) as the type strain, which automatically creates P. bifermentans subsp. bifermentans subsp. nov. JCM 1386 (═ATCC 638 ═DSM 14991 ).
对从生物繁殖大鼠粪便中分离出的专性厌氧、革兰氏阳性、产芽孢细菌PAGU 1678菌株进行了分类学研究。该菌株已被证明能够在葡聚糖硫酸钠诱导的溃疡性结肠炎小鼠模型中加剧病情。基于16S rRNA基因的系统发育分析表明,它与双发酵副梭菌具有高度同源性。为了明确PAGU 1678菌株的正确分类地位,使用双发酵副梭菌PAGU 2008(=JCM 1386)和密切相关的细菌物种解苯副梭菌PAGU 2068(=LMG 28745)进行了比较分类学研究。尽管16S rRNA基因序列相似度很高,但PAGU 1678菌株与双发酵副梭菌PAGU 2008之间的DNA-DNA杂交平均为60.03%,平均核苷酸同一性为96.17%,且显示出不同的基因组序列。在生化方面,PAGU 1678菌株可通过产H₂S与双发酵副梭菌PAGU 2008区分开来。此外,在极性脂质分析中,PAGU 1678菌株的特征是存在两种不同极性的磷脂。此外,PAGU 1678菌株在全细胞蛋白质分析和基质辅助激光解吸/电离飞行时间质谱分析结果上与双发酵副梭菌PAGU 2008不同。基于这些生化和遗传特征,本文提出了双发酵副梭菌的一个新亚种,命名为鼠栖双发酵副梭菌亚种(Paraclostridium bifermentans subsp. muricolitidis subsp. nov.),以PAGU 1678(=CCUG 72489=NBRC 113386)作为模式菌株,这自动产生了双发酵副梭菌双发酵亚种(Paraclostridium bifermentans subsp. bifermentans subsp. nov.)JCM 1386(=ATCC 638=DSM 14991)。