Crespi Sebastian, Drašar Vladimír, Salvà-Serra Francisco, Jaén-Luchoro Daniel, Piñeiro-Iglesias Beatriz, Lindemann Paul Christoffer, Aliaga-Lozano Francisco, Fernández-Juárez Victor, Coll-García Guillem, Moore Edward R B, Bennasar-Figueras Antoni
Biolinea Int.; Calle Sócrates, 4; ES-07007 Palma de Mallorca, Spain.
Public Health Institute Ostrava - National Legionella Reference Laboratory; Masarykovo nam., 16; 682 01 Vyškov, Czech Republic.
Int J Syst Evol Microbiol. 2023 Feb;73(1). doi: 10.1099/ijsem.0.005686.
Two -like strains isolated from hot water distribution systems in 2012 have been characterized phenotypically, biochemically and genomically in terms of DNA relatedness. Both strains, HCPI-6 and EUR-108, exhibited biochemical phenotypic profiles typical of species. Cells were Gram-negative motile rods which grew on BCYEα agar but not on blood agar and displayed phenotypic characteristics typical of the family , including a requirement for l-cysteine and testing catalase positive. Both strains were negative for oxidase, urease, nitrate reduction and hippurate negative, and non-fermentative. The major ubiquinone was Q12 (59.4 % HCPI-6) and the dominant fatty acids were C ω7 (28.4 % HCPI-6, ≈16 % EUR-108), C iso (≈22.5 % and ≈13 %) and C anteiso (19.5 % and ≈23.5 %, respectively). The percent G+C content of genomic DNA was determined to be 39.3 mol %. The 16S rRNA gene, sequence and comparative genome sequence-based analyses (average nucleotide identity, ANI; digital DNA-DNA hybridization, dDDH; and phylogenomic treeing) demonstrated that the strains represent a new species of the genus . The analysis based on the 16S rRNA gene sequences showed that the sequence similarities for both strains ranged from 98.8-90.1 % to other members of the genus. The core genome-based phylogenomic tree (protein-concatemer tree based on concatenation of 418 proteins present in single copy) revealed that these two strains clearly form a separate cluster within the genus . ANI and dDDH values confirmed the distinctiveness of the strains. Based on the genomic, genotypic and phenotypic findings from a polyphasic study, the isolates are considered to represent a single novel species, for which the name sp. nov. is proposed. The type strain is HCPI-6 (=CCUG 75071=CECT 30569).
2012年从热水分配系统中分离出的两株类似菌株已在表型、生化和基因组方面就DNA相关性进行了表征。两株菌株HCPI-6和EUR-108均表现出该物种典型的生化表型特征。细胞为革兰氏阴性运动杆菌,在BCYEα琼脂上生长但不在血琼脂上生长,并表现出该菌科典型的表型特征,包括对L-半胱氨酸的需求和过氧化氢酶检测呈阳性。两株菌株氧化酶、脲酶、硝酸盐还原试验均为阴性,马尿酸盐试验阴性,且不发酵。主要泛醌为Q12(HCPI-6中占59.4%),主要脂肪酸为C ω7(HCPI-6中占28.4%,EUR-108中约占16%)、C异(分别约占22.5%和约13%)和C前异(分别占19.5%和约23.5%)。基因组DNA的G+C含量百分比测定为39.3 mol%。基于16S rRNA基因、序列和基于比较基因组序列的分析(平均核苷酸同一性,ANI;数字DNA-DNA杂交,dDDH;以及系统发育基因组树构建)表明,这些菌株代表了该属的一个新物种。基于16S rRNA基因序列的分析表明,两株菌株与该属其他成员的序列相似性范围为98.8 - 90.1%。基于核心基因组的系统发育基因组树(基于418个单拷贝存在的蛋白质串联构建的蛋白质串联体树)显示,这两株菌株在该属内明显形成一个单独的聚类。ANI和dDDH值证实了这些菌株的独特性。基于多相研究的基因组、基因型和表型结果,这些分离株被认为代表一个单一的新物种,为此提出名称 新种。模式菌株为HCPI-6(=CCUG 75071=CECT 30569)。