Lactobacillus huangpiensis sp. nov. and Lactobacillus laiwuensis sp. nov., isolated from the gut of honeybee (Apis mellifera)

Abstract

Four Gram-stain-positive bacterial strains were isolated from the gut of honeybee (Apis mellifera) in China. These strains were characterized using a polyphasic taxonomic approach. The data demonstrated that three of the four strains represented two novel species of the genus Lactobacillus , strains F306-1T and F551-2T were designated as the type strains. Results of 16S rRNA gene sequence analysis indicated that strains F306-1T, F447 and F551-2T were phylogenetically related to the type strains of Lactobacillus kimbladii and Lactobacillus kullabergensis , having 99.1–99.7 % 16S rRNA gene sequence (about 1400 bp) similarities. The phylogenetic tree based on concatenated pheS, rpoA, gyrB, hsp60, recA, rpoB and tuf sequences (4114 bp) and the phylogenomic tree based on whole genome sequences indicated that strains F306-1T and F447 were most closely related to L. kullabergensis Biut2NT, and strain F551-2T was most closely related to L. kimbladii Hma2NT. Strains F306-1T and F447 shared 99.9 % average nucleotide identity (ANI), 99.7 % digital DNA–DNA hybridization (dDDH) and 99.9 % average amino acid identity (AAI) values, indicating that they belong to the same species. Strain F306-1T exhibited the highest ANI (94.4 %), dDDH (56.7 %) and AAI (94.7 %) values to L. kullabergensis Biut2NT. Strain F551-2T had the highest ANI (94.0 %), dDDH (54.3 %) and AAI (95.8 %) values with L. kimbladii Hma2NT. Acid production from amygdalin, maltose, starch, gentiobiose and turanose, activity of esterase (C4) and α-glucosidase, growth with 3 % NaCl at 37 °C under strict anaerobic condition (on mMRS agar plates), and growth with 1–6% NaCl at 37 °C under aerobic condition (on mMRS agar plates supplemented with 0.05 % cysteine or with 1 % cysteine and 2 % fructose) could differentiate strains F306-1T and F447 from L. kullabergensis DSM 26262T. Acid production from d-glucose, arbutin and gentiobiose, growth with 3 % NaCl at 37 °C under strict anaerobic condition (on mMRS agar plates), and growth at 45 °C under strict anaerobic condition (on mMRS agar plates) could differentiate strain F551-2T from L. kimbladii DSM 26263T. Based upon the data obtained in the present study, two novel species, Lactobacillus huangpiensis sp. nov. and Lactobacillus laiwuensis sp. nov., are proposed and the type strains are F306-1T (=LMG 32144T=JCM 34361T=CCTCC AB 2020300T) and F551-2T (=JCM 34502T=CCTCC AB 2021027T), respectively.

Characterization of Lactic Acid Bacteria Strains Isolated from Algerian Honeybee and Honey and Exploration of Their Potential Probiotic and Functional Features for Human Use

“…The same findings were reported for different or identical phylotypes of the genus Lactobacillus originating from honeybees guts and their food [ 58 , 64 , 65 ]. Recently, three and two species belonging to Apilacobacillus and Lactobacillus , respectively, were described as novel and hosting the Apis mellifera gut [ 10 , 66 , 67 ]. Lactobacillus kimbladii and Lactobacillus kullabergensis , belonging to the Firm-5 clade, were commonly identified as the most numerically abundant phylotype in honey bee guts [ 68 , 69 ].…”

Lactiplantibacillus paraxiangfangensis sp. nov., isolated from traditional Chinese pickle

“…Phenotypic characteristics were investigated as described previously [4, 33]. Resistance to antibiotics was investigated by adding them in mMRS solid medium [34].…”

“…High completeness (85.2-99.4 %) and low contamination (0.6-3.1 %) showed that the genomes used in this study were of high quality (Table S5). Phenotypic characteristics were investigated as described previously [4,33]. Resistance to antibiotics was investigated by adding them in mMRS solid medium [34].…”

Lactobacillus juensis sp. nov. and Lactobacillus rizhaonensis sp. nov., isolated from the gut of honeybee (Apis mellifera)

“…In the genus Lactobacillus, Lactobacillus huangpiensis, Lactobacillus laiwuensis, Lactobacillus helsingborgensis, Lactobacillus melliventris, Lactobacillus kimbladii, Lactobacillus kullabergensis, Lactobacillus apis, Lactobacillus panisapium and Lactobacillus bombicola are associated with bees (Table S1, available in the online version of this article) [1][2][3][4][5], and they form a small phylogenetic branch (L. melliventris clade) [1,6]. All of them are members of Firm5 in honeybee gut microbiota [7].…”

“…Phenotypic features were investigated as described previously [1,8]. Resistance to antibiotics was investigated at four concentrations (5, 50, 100 and 300 µg ml -1 ) by adding antibiotics to mMRS solid medium [29].…”