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Identification of Rare Bacterial Pathogens by 16S rRNA Gene Sequencing and MALDI-TOF MS
Published on: July 11, 2016
Leucobacter faecis sp. nov., a Novel Actinomycete Isolated from Bat Feces of Rousettus and Hipposideros spp
Yuyuan Huang1,2,3, Jing Yang4,5, Qinglian Qin1,2
1Guangxi Key Laboratory of AIDS Prevention and Treatment, School of Public Health, Guangxi Medical University, Nanning, 530021, Guangxi, PR China.
Abstract:
Two novel Gram-stain-positive, non-spore-forming, strictly aerobic, regular rod-shaped bacteria (HY1910T and HY1908) were isolated from bat feces collected from Changshou District of Chongqing City (Altitude: 490 m; N: 30° 02' 15″, E: 107° 07' 4″, September 2011) and Chuxiong Yi Autonomous Prefecture, Yunnan province (Altitude: 1795 m; N: 25°09'10″, E: 102°04'39″, October 2013), respectively. Optimal growth is obtained at 28-30 °C (range, 10-37 °C) on BHI-5% sheep blood plate with pH 7.5 (range, 5.0-10.0) in the presence of 0.5-1.0% NaCl (w/v) [range, 0-5% NaCl (w/v)]. Phylogenetic analysis based on 16S rRNA gene and 458 core genes indicated that strains HY1908 and HY1910T belong to genus Leucobacter, forming a clade with L. salsicius M1-8T (98.6% 16S rRNA gene sequence similarity) and L. exalbidus K-540BT (98.3%). The digital DNA-DNA hybridization (dDDH), average amino acid identity (AAI) and average nucleotide identity (ANI) values of type strain HY1910T with its closest related species were 19.5%-23.5% (dDDH), 58.1%-83.1% (AAI) and 69.4%-80.2% (ANI), falling below the recommended thresholds for species delimitation. In contrast, the dDDH and ANI values between strains HY1908 and HY1910T are 84.0% and 98.1%, respectively, well above the species threshold and suggesting that they belong to the same species. The major fatty acids (> 10.0%) of strain HY1910T were anteiso-C15:0, iso-C16:0 and anteiso-C17:0. Polar lipids included diphosphatidylglycerol, phosphatidylethanolamine, phosphatidylglycerol and two unidentified lipids. Menaquinone 11 (MK-11) was the major respiratory quinone. Based on the combined phylogenetic, chemotaxonomic, and phenotypic evidence, the two strains could be classified as a novel species within the genus Leucobacter, for which the name Leucobacter faecis sp. nov. is proposed (type strain HY1910T=CGMCC 1.19166T=JCM 34880T).
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