Recognition of two subspecies of Lactobacillus amylovorus, with proposal of Lactobacillus amylovorus subsp. animalis subsp. nov. isolated from bovine feces and Lactobacillus amylovorus subsp. amylovorus, and an emended description of Lactobacillus amylovorus

Kenji Yamane; Yasuhiro Tanizawa; Hisami Kobayashi; Tomomi Kamizono; Yoichiro Kojima; Hiroki Takagi; Masanori Tohno

doi:10.51094/jxiv.521

##article.authors##

Kenji Yamane Innovative Animal Production System, University of Tsukuba,Nihon Shokuhin Kako Co., LTD https://orcid.org/0009-0000-8330-7734
Yasuhiro Tanizawa Department of Informatics, National Institute of Genetics https://researchmap.jp/ytani
Hisami Kobayashi Institute of Livestock and Grassland Science, National Agriculture and Food Research Organization https://researchmap.jp/hisamiK
Tomomi Kamizono Institute of Livestock and Grassland Science, National Agriculture and Food Research Organization https://researchmap.jp/t-kamizono
Yoichiro Kojima Institute of Livestock and Grassland Science, National Agriculture and Food Research Organization
Hiroki Takagi Nihon Shokuhin Kako Co., LTD, Research Institute
Masanori Tohno Innovative Animal Production System, University of Tsukuba,Institute of Livestock and Grassland Science, National Agriculture and Food Research Organization,Research Center of Genetic Resources, National Agriculture and Food Research Organization https://researchmap.jp/mtohno

DOI:

https://doi.org/10.51094/jxiv.521

キーワード:

lactic acid bacteria、 bacillota、 lactobacillus amylovorus subsp. animalis、 bovine feves

抄録

Five novel lactic acid bacterial strains (BF125^T, BF186, YK3, YK6, and YK10) were isolated from the fresh feces of Japanese black beef cattle or spent mushroom substrates and characterized using a polyphasic taxonomy method. These strains are rod-shaped, Gram-stain-positive, nonmotile, non-spore-forming, catalase-negative, cytochrome oxidase-negative, facultatively anaerobic, and homofermentative. The cells of BF125^T were 0.5–0.7 µm in width and 3.0–7.0 µm in length. Strain BF125^T did not produce any gas from glucose; both D- and L-lactate were produced as end products of glucose (D/L, 40:60). Growth occurred at a temperature of 30–45°C (optimum, 37°C), pH of 5.0–8.0 (optimum, pH 6.0), and NaCl concentration of 1.0–3.0% (w/v). The GC content of genomic DNA of strain BF125^T was 37.8% (whole-genome analysis). The major fatty acids were C_16:0, C_18:1 w9c, C_19:0 cyclo w8c, and summed feature 10. Strain BF125^T retained high similarity of the 16S rRNA gene to the type strain of Lactobacillus amylovorus (99.93%), and the other isolates were also identified as L. amylovorus based on high 16S rRNA gene similarities. Comparison of the core genomes of L. amylovorus strains, including the five isolates, showed that they could be divided into two clusters, and phenotypic differences in fermentability were observed for D-lactose, salicin, and gentiobiose between these two groups. Digital DNA-DNA hybridization (dDDH) and average nucleotide identity (ANI) analyses showed that both categories were below the thresholds for defining subspecies (maximum dDDH value, 77.2%; maximum ANI value, 96.50%). In light of the physiological, genotypic, and phylogenetic evidence, we propose a novel subspecies of L. amylovorus, L. amylovorus subsp. animalis subsp. nov. Type strain: BF125^T (= MAFF 212522^T = DSM 115528^T). Our results also led to the automatic creation of L. amylovorus subsp. amylovorus subsp. nov. and an emended description of the species L. amylovorus.

利益相反に関する開示

K. Y. and H. T. are employees of Nihon Shokuhin Kako Co., Ltd.. H. K., T. K., and M. T. received research grants from Nihon Shokuhin Kako Co., Ltd.. The authors declare no conflicts of interest.

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