Reclassification of Olsenella gallinarum as Thermophilibacter gallinarum comb. nov. and description of Thermophilibacter immobilis sp. nov., isolated from the mud in a fermentation cellar used for the production of Chinese Luzhou-flavour Baijiu

Abstract

A novel Gram-stain-positive, strictly anaerobic, elliptical, non-motile and non-flagellated bacterium, designed LZLJ-2T, was isolated from the mud in a fermentation cellar used for the production of Chinese Luzhou-flavour Baijiu. Growth occurred at 28–45 °C (optimum, 37 °C), at pH 6.0–7.0 (optimum, pH 6.0) and with concentrations of NaCl up to 2 % (w/v; optimum, 0 %). On the basis of 16S rRNA gene sequence similarity, strain LZLJ-2T belonged to the genus Thermophilibacter and was most closely related to Thermophilibacter mediterraneus Marseille-P3256T (similarity 96.9 %), Olsenella gallinarum ClaCZ62T (similarity 96.6 %) and Thermophilibacter provencensis Marseille-P2912T (similarity 96.4 %). In addition, strain LZLJ-2T had high similarity to the genus Olsenella , including Olsenella profusa DSM 13989T (similarity 94.9 %), Olsenella umbonata DSM 22620T (similarity 94.9 %), Olsenella uli ATCC 49627T (similarity 94.22 %), Tractidigestivibacter scatoligenes DSM 28304T (similarity 93.9 %) and Paratractidigestivibacter faecalis KCTC 15699T (similarity 93.25 %). Comparative genome analysis showed that orthoANI values between strain LZLJ-2T and Thermophilibacter mediterraneus Marseille-P3256T, Olsenella gallinarum ClaCZ62T, Thermophilibacter provencensis Marseille-P2912T, Olsenella profusa DSM 13989T, Olsenella umbonata DSM 22620T, Olsenella uli ATCC 49627T, Tractidigestivibacter scatoligenes DSM 28304T and Paratractidigestivibacter faecalis KCTC 15699T were 78.68, 78.99, 78.29, 73.40, 74.00, 74.30, 75.08 and 77.23 %, and the genome-to-genome distance values were respectively 22.3, 22.5, 22.4, 19.6, 20.5, 19.7, 20.5 and 21.5 %. The genomic DNA G+C content of strain LZLJ-2T was 65.21 mol%. The predominant cellular fatty acids (>10 %) of strain LZLJ-2T were C18 : 1 cis 9 (33.7 %), C14 : 0 (22.0 %) and C18 : 1 cis 9 DMA (13.5 %). d-Glucose, sucrose, mannose, maltose, lactose (weak), salicin, glycerol (weak), cellobiose and trehalose (weak) could be used by strain LZLJ-2T as sole carbon sources. Enzyme activity results showed positive reactions with valine arylamidase, leucine arylamidase, crystine arylamidase, acid phosphatase, alkaline phosphatase, esterase (C4) (weakly positive), naphthol-AS-BI-phosphohydrolase, α-glucosidase and β-glucosidase. The major end products of glucose fermentation were lactic acid and acetic acid. It produced skatole from indole acetic acid, and produced p-cresol from modified peptone–yeast extract medium with glucose. Based on the 16S rRNA gene trees as well as the genome core gene tree, it is suggested that Olsenella gallinarum are transferred to genus Thermophilibacter as Thermophilibacter gallinarum comb. nov. Based on phenotypic, genotypic and phylogenetic data, strain LZLJ-2T is considered to represent a novel species of the genus Thermophilibacter , for which the name Thermophilibacter immobilis sp. nov. is proposed. The type strain is LZLJ-2T (=KCTC 25162T=JCM 34224T).

From pollen to putrid: Comparative metagenomics reveals how microbiomes support dietary specialization in vulture bees

“…and Acinetobacter sp. but also other microbes that are found on vertebrate bodies or mud such as Salmonella enterica , Escherichia coli , Thermophillibacter immobilis , Pseudomonas sp., Olsenella uli , Paralysiella testudinis , Comamonas sp., and Klebsiella sp., several of which are considered opportunistic human pathogens (Busse et al., 2021; Lu et al., 2021; Ryan et al., 2022). Likewise, facultative necrophages have some bacteria known to be linked to pollinivorous diets such as Prevotella sp.…”

“…The latter two, as well as Gilliamella , Bombella (formerly Parasaccharibacter ), and S accharibacter , were contributing several differentially more abundant genes in the pollinivores versus the obligate necrophage (Figure 2). The facultatively necrophagous bees had a high abundance of anaerobic microbes known to be environmentally acquired such as Pectinatus (involved in beer spoilage) in addition to microbes associated with vertebrates ( Paralysiella testudinis – only ever isolated from a turtle cloaca) and even mud ( Thermophilibacter immobilis ) (Busse et al., 2021; Lu et al., 2021; Rodríguez‐Saavedra et al., 2021). In addition to the environmental microbes, the facultative necrophages also maintain a high abundance of the corbiculate and Meliponini core such as Gilliamella and Snodgrassella , Apibacter , and Commensalibacter .…”

Systematic Review of Actinomycetes in the Baijiu Fermentation Microbiome

“…Arthrobacter protophormiae was isolated using inorganic salt medium from 20-year-old PM [ 38 ]. Streptomyces sampsonii and Streptomyces rutgersensis were separated from the in situ medium and the Thermophilibacter gallinarum R2A medium from the PM [ 41 , 42 ]. Three genera ( Streptomyces , Nocardiopsis , and Massilia ) and 26 species were isolated from the fermentations using modified Gao’s No.1 medium [ 35 ].…”

“…Aside from enzymes, actinomycetes also produce many metabolites, including aroma compounds, antibiotics, and off-odor compounds ( Table 4 ). Thermophilibacter gallinarum can convert glucose into lactic acid and acetic acid [ 41 ]. Actinomycetes also have the potential to produce butyric acid, thereby providing the precursor for ethyl butyrate and enriching the flavor of the Baijiu [ 143 ].…”