Journal: Microorganisms

Article Title: Enzymatic Adaptation of Bifidobacterium bifidum to Host Glycans, Viewed from Glycoside Hydrolyases and Carbohydrate-Binding Modules

doi: 10.3390/microorganisms8040481

Figure Lengend Snippet: Enzymes involved in the degradation and assimilation of human milk oligosaccharides (HMOs) and mucin O -glycans by B. bifidum . The sugars are depicted according to the nomenclature committee of the Consortium for Functional Glycomics ( http://www.functionalglycomics.org/static/index.shtml ).

Article Snippet: As visualized in , B. bifidum [strains PRL2010, LMG 13195 (= JCM 7004), and TMC 3115] has 14 extracellular GHs involved in HMO/mucin O -glycan degradation, as predicted in silico, whereas B. infantis ATCC 15697 (= JCM 1222) and B. breve UCC2003 are equipped with the intracellular counterpart enzymes (e.g., GH33 sialidase and GH95 1,2-α- l -fucosidase).

Techniques: Functional Assay

Journal: Microorganisms

Article Title: Enzymatic Adaptation of Bifidobacterium bifidum to Host Glycans, Viewed from Glycoside Hydrolyases and Carbohydrate-Binding Modules

doi: 10.3390/microorganisms8040481

Figure Lengend Snippet: B. bifidum -mediated cross-feeding supports the growth of B. longum on HMO- and mucin-containing media. ( A – D ): The growth of B. longum 105-A in a basal medium supplemented with 1% human milk oligosaccharides (HMOs) ( A , B ) or 1% porcine gastric mucin ( C , D ) in the absence ( A , C ) and presence ( B , D ) of B. bifidum JCM 1254. The wild-type B. longum 105-A strain carrying the chloramphenicol ( A , B ) or spectinomycin ( C – F ) resistance gene on a plasmid (pBFS38 or pBFO2, respectively) was used for monitoring the growth. Colony-forming units (CFU) of B. longum 105-A were determined by spreading the serial dilution of the cultures on the agar plates containing the antibiotics (closed circles), while the CFU of total Bifidobacterium was determined using the agar plates without antibiotics (open circles). The data used in ( A ) was obtained from our previous study . ( E , F ): The growth competition between wild-type and Δ gltA mutant strains of B. longum 105-A in the presence of B. bifidum JCM 1254. Lactose (Lac), HMOs, or PGM was used as the sole carbon source. The ratio of the growth on HMOs ( E ) or PGM ( F ) that was normalized by that on Lac was compared. The Δ gltA mutant was transformed with pBFS38 carrying the chloramphenicol resistance gene, and CFU was determined on agar plates containing the antibiotics.

Article Snippet: As visualized in , B. bifidum [strains PRL2010, LMG 13195 (= JCM 7004), and TMC 3115] has 14 extracellular GHs involved in HMO/mucin O -glycan degradation, as predicted in silico, whereas B. infantis ATCC 15697 (= JCM 1222) and B. breve UCC2003 are equipped with the intracellular counterpart enzymes (e.g., GH33 sialidase and GH95 1,2-α- l -fucosidase).

Techniques: Plasmid Preparation, Serial Dilution, Mutagenesis, Transformation Assay

Journal: Frontiers in Microbiology

Article Title: Probiotic Mixture Golden Bifido Prevents Neonatal Escherichia coli K1 Translocation via Enhancing Intestinal Defense

doi: 10.3389/fmicb.2017.01798

Figure Lengend Snippet: Lactobacillus bulgaricus, Bifidobacterium , and Streptococcus thermophilus (LBS) alleviates mucin degradation during Escherichia coli K1 infection. HT-29 cells were pre-treated with or without LBS before E. coli K1 infection. Cells treated with PBS or LBS alone were served as controls. Cellular proteins were extracted for (A) periodic acid-Schiff (PAS) assay and (C) western blot analysis. (B) HT-29 monolayers were stained with PAS and observed under light microscope (100× magnification). (D) Immunofluorescence of MUC2 was obtained by fluorescence microscopy (200× magnification), green staining represents the MUC2, the cell nuclei were stained with Hoechst 33342 (blue). Results are represented as mean ± SD. ∗ P < 0.05, ∗∗∗ P < 0.001.

Article Snippet: The probiotics used in this study was obtained from Live Combined bifidobacterium and lactobacillus Tablets [Trade Name: Golden Bifid, Inner Mongolia Shuangqi Pharmaceutical Co. Ltd., 0.5 g/tablet, no less than 50 million live Bifidobacterium (ATCC 15697), five million live L. bulgaricus (ATCC 11842) and S. thermophilus (ATCC 19987)].

Techniques: Infection, PAS Assay, Western Blot, Staining, Light Microscopy, Immunofluorescence, Fluorescence, Microscopy

Journal: Frontiers in Microbiology

Article Title: Probiotic Mixture Golden Bifido Prevents Neonatal Escherichia coli K1 Translocation via Enhancing Intestinal Defense

doi: 10.3389/fmicb.2017.01798

Figure Lengend Snippet: Lactobacillus bulgaricus, Bifidobacterium , and Streptococcus thermophilus inhibits adhesion and invasion of E. coli K1 to HT-29 cells. HT-29 cells were cultured in 24-well plate and pre-treated with various doses of LBS for 3 h before adding E. coli K1 or co-incubated with LBS plus E. coli K1. Adhesion and invasion assays were carried out as described in section “Materials and Methods.” The numbers of associated bacteria (A,C) and intracellular bacteria (B,D) were determined. Results are represented as mean ± SD. ∗ P < 0.05, ∗∗ P < 0.01.

Article Snippet: The probiotics used in this study was obtained from Live Combined bifidobacterium and lactobacillus Tablets [Trade Name: Golden Bifid, Inner Mongolia Shuangqi Pharmaceutical Co. Ltd., 0.5 g/tablet, no less than 50 million live Bifidobacterium (ATCC 15697), five million live L. bulgaricus (ATCC 11842) and S. thermophilus (ATCC 19987)].

Techniques: Cell Culture, Incubation, Bacteria

Journal: Frontiers in Microbiology

Article Title: Probiotic Mixture Golden Bifido Prevents Neonatal Escherichia coli K1 Translocation via Enhancing Intestinal Defense

doi: 10.3389/fmicb.2017.01798

Figure Lengend Snippet: Lactobacillus bulgaricus, Bifidobacterium , and Streptococcus thermophilus prevents E. coli K1-induced disruption of intestinal integrity. HT-29 cells were cultured on the upper chamber of the Transwell insert and pre-treated with or without LBS before E. coli K1 infection. Cells treated with PBS or LBS alone were served as controls. After infection, HRP was added to the upper chamber of Transwell for 1 h. Bacteria and HRP translocated from the upper chamber to the lower chamber were quantified as described in section “Materials and Methods.” (A) The OD 450 value of HRP. (B) The number of E. coli K1 CFU, which penetrated across the HT-29 monolayer. Results are represented as mean ± SD. ∗ P < 0.05, ∗∗∗ P < 0.001. (C) HT-29 cells were treated as described in Figure , proteins of each group were isolated for western blotting. The expressions of ZO-1 and occludin was determined, β-Actin band was used as an indicator of protein loading.

Article Snippet: The probiotics used in this study was obtained from Live Combined bifidobacterium and lactobacillus Tablets [Trade Name: Golden Bifid, Inner Mongolia Shuangqi Pharmaceutical Co. Ltd., 0.5 g/tablet, no less than 50 million live Bifidobacterium (ATCC 15697), five million live L. bulgaricus (ATCC 11842) and S. thermophilus (ATCC 19987)].

Techniques: Disruption, Cell Culture, Infection, Bacteria, Isolation, Western Blot