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Difco in on brain heart infusion bhi
In On Brain Heart Infusion Bhi, supplied by Difco, used in various techniques. Bioz Stars score: 78/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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In Vivo:

Article Title: Contribution of Individual Ebp Pilus Subunits of Enterococcus faecalis OG1RF to Pilus Biogenesis, Biofilm Formation and Urinary Tract Infection
Article Snippet: .. E. faecalis strains were routinely grown in/on brain heart infusion (BHI) (Difco Laboratories, Detroit, MI) broth/agar or, for some experiments (see below), in tryptic soy broth supplemented with 0.25% glucose (TSBG) and, for in vivo experiments, in BHI supplemented with 40% horse serum (BHIS). .. E. coli strains were grown in Luria-Bertani media (Difco Laboratories).

Plasmid Preparation:

Article Title: Contribution of Individual Ebp Pilus Subunits of Enterococcus faecalis OG1RF to Pilus Biogenesis, Biofilm Formation and Urinary Tract Infection
Article Snippet: Plasmid constructs were given pTEX numbers and their host strains corresponding TX numbers. .. E. faecalis strains were routinely grown in/on brain heart infusion (BHI) (Difco Laboratories, Detroit, MI) broth/agar or, for some experiments (see below), in tryptic soy broth supplemented with 0.25% glucose (TSBG) and, for in vivo experiments, in BHI supplemented with 40% horse serum (BHIS).

Article Title: Characterization of the ebpfm pilus-encoding operon of Enterococcus faecium and its role in biofilm formation and virulence in a murine model of urinary tract infection
Article Snippet: Plasmid constructs were given pTEX numbers and their host strains corresponding TX numbers. .. E. faecium strains were routinely grown in/on brain heart infusion (BHI) (Difco Laboratories, Detroit, MI) broth/agar or, for some experiments (see below), in tryptic soy broth supplemented with 0.25% glucose (TSBG) or on trypticase soy agar supplemented with 5% sheep blood (Becton Dickinson, Franklin Lakes, NJ), and E. coli in Luria-Bertani media (Difco Laboratories) at 37°C.

Article Title: Characterization of the ebpfm pilus-encoding operon of Enterococcus faecium and its role in biofilm formation and virulence in a murine model of urinary tract infection
Article Snippet: Plasmid constructs were given pTEX numbers and their host strains corresponding TX numbers. .. E. faecium strains were routinely grown in/on brain heart infusion (BHI) (Difco Laboratories, Detroit, MI) broth/agar or, for some experiments (see below), in tryptic soy broth supplemented with 0.25% glucose (TSBG) or on trypticase soy agar supplemented with 5% sheep blood (Becton Dickinson, Franklin Lakes, NJ), and E. coli in Luria-Bertani media (Difco Laboratories) at 37°C.

Construct:

Article Title: Contribution of Individual Ebp Pilus Subunits of Enterococcus faecalis OG1RF to Pilus Biogenesis, Biofilm Formation and Urinary Tract Infection
Article Snippet: Plasmid constructs were given pTEX numbers and their host strains corresponding TX numbers. .. E. faecalis strains were routinely grown in/on brain heart infusion (BHI) (Difco Laboratories, Detroit, MI) broth/agar or, for some experiments (see below), in tryptic soy broth supplemented with 0.25% glucose (TSBG) and, for in vivo experiments, in BHI supplemented with 40% horse serum (BHIS).

Article Title: Characterization of the ebpfm pilus-encoding operon of Enterococcus faecium and its role in biofilm formation and virulence in a murine model of urinary tract infection
Article Snippet: Plasmid constructs were given pTEX numbers and their host strains corresponding TX numbers. .. E. faecium strains were routinely grown in/on brain heart infusion (BHI) (Difco Laboratories, Detroit, MI) broth/agar or, for some experiments (see below), in tryptic soy broth supplemented with 0.25% glucose (TSBG) or on trypticase soy agar supplemented with 5% sheep blood (Becton Dickinson, Franklin Lakes, NJ), and E. coli in Luria-Bertani media (Difco Laboratories) at 37°C.

Article Title: Characterization of the ebpfm pilus-encoding operon of Enterococcus faecium and its role in biofilm formation and virulence in a murine model of urinary tract infection
Article Snippet: Plasmid constructs were given pTEX numbers and their host strains corresponding TX numbers. .. E. faecium strains were routinely grown in/on brain heart infusion (BHI) (Difco Laboratories, Detroit, MI) broth/agar or, for some experiments (see below), in tryptic soy broth supplemented with 0.25% glucose (TSBG) or on trypticase soy agar supplemented with 5% sheep blood (Becton Dickinson, Franklin Lakes, NJ), and E. coli in Luria-Bertani media (Difco Laboratories) at 37°C.

Selection:

Article Title: Characterization of the ebpfm pilus-encoding operon of Enterococcus faecium and its role in biofilm formation and virulence in a murine model of urinary tract infection
Article Snippet: E. faecium strains were routinely grown in/on brain heart infusion (BHI) (Difco Laboratories, Detroit, MI) broth/agar or, for some experiments (see below), in tryptic soy broth supplemented with 0.25% glucose (TSBG) or on trypticase soy agar supplemented with 5% sheep blood (Becton Dickinson, Franklin Lakes, NJ), and E. coli in Luria-Bertani media (Difco Laboratories) at 37°C. .. With E. faecium TX82, gentamicin (Sigma Chemical Co., St. Louis, MO) was used at 125 µg/mL (150 µg/mL for selection of transformants) and chloramphenicol at 10 µg/mL and, with E. coli , gentamicin was used at 25 µg/mL.

Article Title: Characterization of the ebpfm pilus-encoding operon of Enterococcus faecium and its role in biofilm formation and virulence in a murine model of urinary tract infection
Article Snippet: E. faecium strains were routinely grown in/on brain heart infusion (BHI) (Difco Laboratories, Detroit, MI) broth/agar or, for some experiments (see below), in tryptic soy broth supplemented with 0.25% glucose (TSBG) or on trypticase soy agar supplemented with 5% sheep blood (Becton Dickinson, Franklin Lakes, NJ), and E. coli in Luria-Bertani media (Difco Laboratories) at 37°C. .. With E. faecium TX82, gentamicin (Sigma Chemical Co., St. Louis, MO) was used at 125 μg/mL (150 μg/mL for selection of transformants) and chloramphenicol at 10 μg/mL and, with E. coli , gentamicin was used at 25 μg/mL.

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  • 99
    Difco brain heart infusion bhi broth
    Western blot analysis of the urea-activated fraction using antibodies against RgpB. All lanes contained 20 μg of acetone (37.5%)-precipitated proteins from the supernatant fractions of P. <t>gingivalis</t> FLL92 cultures grown in <t>BHI</t> medium at exponential growth phases. The membrane was reacted with antiserum raised in rabbits against RgpB. Lanes: 1, FLL92 exponential-growth-phase extracellular proteins; 2, FLL92 exponential-growth-phase extracellular proteins treated with urea (see the text).
    Brain Heart Infusion Bhi Broth, supplied by Difco, used in various techniques. Bioz Stars score: 99/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/brain heart infusion bhi broth/product/Difco
    Average 99 stars, based on 3 article reviews
    Price from $9.99 to $1999.99
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    96
    Difco brain heart infusion bhi medium
    Beta-lactam protection in a polymicrobial biofilm. Stationary biofilms of H. influenzae Rd and/or M. <t>catarrhalis</t> were established on chamber slides for 24 h and treated with 100 µg/ml ampicillin or ampicillin with 25 µg/ml clavulanate for an additional 24 h. Biofilms were resuspended in sterile PBS, serially diluted, and plated on sBHI medium plus clarithromycin and <t>BHI</t> medium plates for enumeration of viable H. influenzae Rd and M. catarrhalis bacteria, respectively. Data are represented as means ± SEM. *, P
    Brain Heart Infusion Bhi Medium, supplied by Difco, used in various techniques. Bioz Stars score: 96/100, based on 34 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
    https://www.bioz.com/result/brain heart infusion bhi medium/product/Difco
    Average 96 stars, based on 34 article reviews
    Price from $9.99 to $1999.99
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    84
    Difco brain heart infusion medium bhi
    pPD1 enhances EF competition for an intestinal niche (a) Bacteriocin assay by the soft agar method with EF r +pPD1::Δ bacAB, bacA-E +, EF r +pPD1 and EF r +pPD1::Δ bacAB spotted on a lawn of susceptible EF. Mice (N=5/group) were given EF r or EF r +pPD1 or EF r +pPD1::Δ bacAB or sterile drinking water for 14 days, at which time all mice were given sterile water. (b) One week and (c) four weeks after withdrawal of EF from drinking water, fecal samples were collected and abundance of total enterococci was determined by enumeration on m-Enterococcus selective agar <t>(Ent</t> agar, ○). Lab strain of EF was enumerated using Ent agar with rifampicin (Rf) (○), or <t>BHI</t> agar with rifampicin (○). (d and e and Fig. 1e ) At the end of week four, animals were euthanized and abundance of EF was determined in distal small intestine (DSI) and large intestine (LI). The results shown are representative of two biologically independent experiments. Horizontal lines indicate geometric mean. Each symbol represents an individual animal.
    Brain Heart Infusion Medium Bhi, supplied by Difco, used in various techniques. Bioz Stars score: 84/100, based on 3 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Western blot analysis of the urea-activated fraction using antibodies against RgpB. All lanes contained 20 μg of acetone (37.5%)-precipitated proteins from the supernatant fractions of P. gingivalis FLL92 cultures grown in BHI medium at exponential growth phases. The membrane was reacted with antiserum raised in rabbits against RgpB. Lanes: 1, FLL92 exponential-growth-phase extracellular proteins; 2, FLL92 exponential-growth-phase extracellular proteins treated with urea (see the text).

    Journal: Infection and Immunity

    Article Title: Gingipain RgpB Is Excreted as a Proenzyme in the vimA-Defective Mutant Porphyromonas gingivalis FLL92

    doi: 10.1128/IAI.71.7.3740-3747.2003

    Figure Lengend Snippet: Western blot analysis of the urea-activated fraction using antibodies against RgpB. All lanes contained 20 μg of acetone (37.5%)-precipitated proteins from the supernatant fractions of P. gingivalis FLL92 cultures grown in BHI medium at exponential growth phases. The membrane was reacted with antiserum raised in rabbits against RgpB. Lanes: 1, FLL92 exponential-growth-phase extracellular proteins; 2, FLL92 exponential-growth-phase extracellular proteins treated with urea (see the text).

    Article Snippet: P. gingivalis W83 and P. gingivalis FLL92 ( vimA :: ermF- ermAM ) were grown in brain heart infusion (BHI) broth (Difco Laboratories, Detroit, Mich.) supplemented with hemin (5 μg/ml), vitamin K (0.5 μg/ml), and cysteine (1%).

    Techniques: Western Blot

    Protease activation in the extracellular protein fraction from P. gingivalis FLL92. P. gingivalis was grown to late log phase (OD 660 of 0.8) in 1 liter of BHI broth supplemented with hemin and vitamin K. Acetone (37.5%)-precipitated proteins were mixed with 8 M urea and incubated at 4°C for 1 h. The urea was slowly removed from the mixture by centrifugation (10,000 × g ). The results shown are representative of four independent experiments.

    Journal: Infection and Immunity

    Article Title: Gingipain RgpB Is Excreted as a Proenzyme in the vimA-Defective Mutant Porphyromonas gingivalis FLL92

    doi: 10.1128/IAI.71.7.3740-3747.2003

    Figure Lengend Snippet: Protease activation in the extracellular protein fraction from P. gingivalis FLL92. P. gingivalis was grown to late log phase (OD 660 of 0.8) in 1 liter of BHI broth supplemented with hemin and vitamin K. Acetone (37.5%)-precipitated proteins were mixed with 8 M urea and incubated at 4°C for 1 h. The urea was slowly removed from the mixture by centrifugation (10,000 × g ). The results shown are representative of four independent experiments.

    Article Snippet: P. gingivalis W83 and P. gingivalis FLL92 ( vimA :: ermF- ermAM ) were grown in brain heart infusion (BHI) broth (Difco Laboratories, Detroit, Mich.) supplemented with hemin (5 μg/ml), vitamin K (0.5 μg/ml), and cysteine (1%).

    Techniques: Activation Assay, Incubation, Centrifugation

    Detection of proteolytic activity in casein-conjugated polyacrylamide gel. All lanes contained 20 μg of acetone (37.5%)-precipitated proteins from the supernatant fractions of cultures in BHI medium at exponential growth phases. Samples were solubilized at room temperature for 10 min in a Tris-glycine SDS sample buffer (Invitrogen) prior to electrophoresis. Lanes: 1, W83; 2, FLL92. The arrow indicates a 48-kDa proteolytic band present in the FLL92 exponential-growth-phase extracellular proteins.

    Journal: Infection and Immunity

    Article Title: Gingipain RgpB Is Excreted as a Proenzyme in the vimA-Defective Mutant Porphyromonas gingivalis FLL92

    doi: 10.1128/IAI.71.7.3740-3747.2003

    Figure Lengend Snippet: Detection of proteolytic activity in casein-conjugated polyacrylamide gel. All lanes contained 20 μg of acetone (37.5%)-precipitated proteins from the supernatant fractions of cultures in BHI medium at exponential growth phases. Samples were solubilized at room temperature for 10 min in a Tris-glycine SDS sample buffer (Invitrogen) prior to electrophoresis. Lanes: 1, W83; 2, FLL92. The arrow indicates a 48-kDa proteolytic band present in the FLL92 exponential-growth-phase extracellular proteins.

    Article Snippet: P. gingivalis W83 and P. gingivalis FLL92 ( vimA :: ermF- ermAM ) were grown in brain heart infusion (BHI) broth (Difco Laboratories, Detroit, Mich.) supplemented with hemin (5 μg/ml), vitamin K (0.5 μg/ml), and cysteine (1%).

    Techniques: Activity Assay, Electrophoresis

    SDS-PAGE of acetone-precipitated P. gingivalis extracellular proteins. Proteins in the supernatant fractions of cultures at different growth phases in BHI medium were precipitated with 37.5% (A) or 60% (B) acetone. The fractions were solubilized at 72°C in reducing buffer. NuPAGE bis-Tris gels (4 to 12%) were stained with Simply Blue Safe stain. Lanes: 1, W83 at exponential phase; 2, FLL92 at exponential phase; 3, W83 at stationary phase; and 4, FLL92 at stationary phase. The molecular mass markers (in kilodaltons) are indicated on the left. Each lane contains 20 μg of protein. It is noteworthy that the 64-kDa band was observed in abundance when 37.5% acetone instead of 60% acetone was used for the precipitation.

    Journal: Infection and Immunity

    Article Title: Gingipain RgpB Is Excreted as a Proenzyme in the vimA-Defective Mutant Porphyromonas gingivalis FLL92

    doi: 10.1128/IAI.71.7.3740-3747.2003

    Figure Lengend Snippet: SDS-PAGE of acetone-precipitated P. gingivalis extracellular proteins. Proteins in the supernatant fractions of cultures at different growth phases in BHI medium were precipitated with 37.5% (A) or 60% (B) acetone. The fractions were solubilized at 72°C in reducing buffer. NuPAGE bis-Tris gels (4 to 12%) were stained with Simply Blue Safe stain. Lanes: 1, W83 at exponential phase; 2, FLL92 at exponential phase; 3, W83 at stationary phase; and 4, FLL92 at stationary phase. The molecular mass markers (in kilodaltons) are indicated on the left. Each lane contains 20 μg of protein. It is noteworthy that the 64-kDa band was observed in abundance when 37.5% acetone instead of 60% acetone was used for the precipitation.

    Article Snippet: P. gingivalis W83 and P. gingivalis FLL92 ( vimA :: ermF- ermAM ) were grown in brain heart infusion (BHI) broth (Difco Laboratories, Detroit, Mich.) supplemented with hemin (5 μg/ml), vitamin K (0.5 μg/ml), and cysteine (1%).

    Techniques: SDS Page, Staining

    Western immunoblot analysis of the extracellular proteins from P. gingivalis by using specific anti-Rgp and anti-Kgp antibodies as probes. All lanes contained 20 μg of acetone (37.5%)-precipitated proteins from the supernatant fractions of cultures in BHI medium at different growth phases. The membrane was reacted with antiserum raised in rabbits or chickens against the Arg-X- and Lys-X-specific protease from P. gingivalis . The secondary antibody was goat anti-rabbit or anti-chicken immunoglobulin G-horseradish peroxidase conjugate.(Zymed Laboratories Inc.). Reactions were done with rabbit anti-RgpB (A), rabbit anti-RgpA (B), and chicken anti-Kgp (C). Lanes: 1, W83 at exponential phase; 2, FLL92 at exponential phase; 3, W83 at stationary phase; 4, FLL92 at stationary phase. It is noteworthy that the 64-kDa band was identified as the RgpB proenzyme by mass spectrometry (see the text).

    Journal: Infection and Immunity

    Article Title: Gingipain RgpB Is Excreted as a Proenzyme in the vimA-Defective Mutant Porphyromonas gingivalis FLL92

    doi: 10.1128/IAI.71.7.3740-3747.2003

    Figure Lengend Snippet: Western immunoblot analysis of the extracellular proteins from P. gingivalis by using specific anti-Rgp and anti-Kgp antibodies as probes. All lanes contained 20 μg of acetone (37.5%)-precipitated proteins from the supernatant fractions of cultures in BHI medium at different growth phases. The membrane was reacted with antiserum raised in rabbits or chickens against the Arg-X- and Lys-X-specific protease from P. gingivalis . The secondary antibody was goat anti-rabbit or anti-chicken immunoglobulin G-horseradish peroxidase conjugate.(Zymed Laboratories Inc.). Reactions were done with rabbit anti-RgpB (A), rabbit anti-RgpA (B), and chicken anti-Kgp (C). Lanes: 1, W83 at exponential phase; 2, FLL92 at exponential phase; 3, W83 at stationary phase; 4, FLL92 at stationary phase. It is noteworthy that the 64-kDa band was identified as the RgpB proenzyme by mass spectrometry (see the text).

    Article Snippet: P. gingivalis W83 and P. gingivalis FLL92 ( vimA :: ermF- ermAM ) were grown in brain heart infusion (BHI) broth (Difco Laboratories, Detroit, Mich.) supplemented with hemin (5 μg/ml), vitamin K (0.5 μg/ml), and cysteine (1%).

    Techniques: Western Blot, Mass Spectrometry

    Sensitivity of P. gingivalis W83, FLL32, FLL455, and FLL456 to UV irradiation. Dilutions of fresh cultures of the P. gingivalis strains were grown anaerobically to exponential phase in BHI broth from overnight cultures and spread on BHI agar. The cells

    Journal: Journal of Bacteriology

    Article Title: Nitric Oxide Stress Resistance in Porphyromonas gingivalis Is Mediated by a Putative Hydroxylamine Reductase

    doi: 10.1128/JB.06457-11

    Figure Lengend Snippet: Sensitivity of P. gingivalis W83, FLL32, FLL455, and FLL456 to UV irradiation. Dilutions of fresh cultures of the P. gingivalis strains were grown anaerobically to exponential phase in BHI broth from overnight cultures and spread on BHI agar. The cells

    Article Snippet: P. gingivalis strains were grown in brain heart infusion (BHI) broth supplemented with 0.5% yeast extract (Difco Laboratories, Detroit, MI), hemin (5 μg ml−1 ), vitamin K (0.5 μg ml−1 ), and cysteine (0.1%; Sigma-Aldrich, St. Louis, MO).

    Techniques: Irradiation

    Sensitivity of P. gingivalis W83, FLL455 and FLL456 to single NO stress. P. gingivalis strains were grown anaerobically to early exponential phase in BHI broth at 37°C, DEA NONOate (NO) was added to the cultures, and the cultures were further

    Journal: Journal of Bacteriology

    Article Title: Nitric Oxide Stress Resistance in Porphyromonas gingivalis Is Mediated by a Putative Hydroxylamine Reductase

    doi: 10.1128/JB.06457-11

    Figure Lengend Snippet: Sensitivity of P. gingivalis W83, FLL455 and FLL456 to single NO stress. P. gingivalis strains were grown anaerobically to early exponential phase in BHI broth at 37°C, DEA NONOate (NO) was added to the cultures, and the cultures were further

    Article Snippet: P. gingivalis strains were grown in brain heart infusion (BHI) broth supplemented with 0.5% yeast extract (Difco Laboratories, Detroit, MI), hemin (5 μg ml−1 ), vitamin K (0.5 μg ml−1 ), and cysteine (0.1%; Sigma-Aldrich, St. Louis, MO).

    Techniques:

    Sensitivity of P. gingivalis W83, FLL455, and complemented (comp) FLL455 to NO stress. P. gingivalis strains were grown anaerobically to early exponential phase in BHI broth at 37°C, DEA NONOate (NO) was added to the cultures, and the cultures

    Journal: Journal of Bacteriology

    Article Title: Nitric Oxide Stress Resistance in Porphyromonas gingivalis Is Mediated by a Putative Hydroxylamine Reductase

    doi: 10.1128/JB.06457-11

    Figure Lengend Snippet: Sensitivity of P. gingivalis W83, FLL455, and complemented (comp) FLL455 to NO stress. P. gingivalis strains were grown anaerobically to early exponential phase in BHI broth at 37°C, DEA NONOate (NO) was added to the cultures, and the cultures

    Article Snippet: P. gingivalis strains were grown in brain heart infusion (BHI) broth supplemented with 0.5% yeast extract (Difco Laboratories, Detroit, MI), hemin (5 μg ml−1 ), vitamin K (0.5 μg ml−1 ), and cysteine (0.1%; Sigma-Aldrich, St. Louis, MO).

    Techniques:

    (A) Sensitivity of P. gingivalis W83 to single NO stress. P. gingivalis W83 strain was grown anaerobically to early exponential phase in BHI broth at 37°C, DEA NONOate (NO) or NaOH was added to the cultures, and the cultures were further incubated

    Journal: Journal of Bacteriology

    Article Title: Nitric Oxide Stress Resistance in Porphyromonas gingivalis Is Mediated by a Putative Hydroxylamine Reductase

    doi: 10.1128/JB.06457-11

    Figure Lengend Snippet: (A) Sensitivity of P. gingivalis W83 to single NO stress. P. gingivalis W83 strain was grown anaerobically to early exponential phase in BHI broth at 37°C, DEA NONOate (NO) or NaOH was added to the cultures, and the cultures were further incubated

    Article Snippet: P. gingivalis strains were grown in brain heart infusion (BHI) broth supplemented with 0.5% yeast extract (Difco Laboratories, Detroit, MI), hemin (5 μg ml−1 ), vitamin K (0.5 μg ml−1 ), and cysteine (0.1%; Sigma-Aldrich, St. Louis, MO).

    Techniques: Incubation

    Glutamate concentration in P. gingivalis W83, FLL455, and complemented FLL455. P. gingivalis strains were grown anaerobically to early exponential phase in BHI broth at 37°C, and cultures were either left untreated or treated with the indicated

    Journal: Journal of Bacteriology

    Article Title: Nitric Oxide Stress Resistance in Porphyromonas gingivalis Is Mediated by a Putative Hydroxylamine Reductase

    doi: 10.1128/JB.06457-11

    Figure Lengend Snippet: Glutamate concentration in P. gingivalis W83, FLL455, and complemented FLL455. P. gingivalis strains were grown anaerobically to early exponential phase in BHI broth at 37°C, and cultures were either left untreated or treated with the indicated

    Article Snippet: P. gingivalis strains were grown in brain heart infusion (BHI) broth supplemented with 0.5% yeast extract (Difco Laboratories, Detroit, MI), hemin (5 μg ml−1 ), vitamin K (0.5 μg ml−1 ), and cysteine (0.1%; Sigma-Aldrich, St. Louis, MO).

    Techniques: Concentration Assay

    Sensitivity of P. gingivalis W83, FLL455, and FLL456 to H 2 O 2 stress. P. gingivalis strains were grown anaerobically to early exponential phase in BHI broth at 37°C, H 2 O 2 was added to the cultures, and the cultures further incubated for 24 h. Each

    Journal: Journal of Bacteriology

    Article Title: Nitric Oxide Stress Resistance in Porphyromonas gingivalis Is Mediated by a Putative Hydroxylamine Reductase

    doi: 10.1128/JB.06457-11

    Figure Lengend Snippet: Sensitivity of P. gingivalis W83, FLL455, and FLL456 to H 2 O 2 stress. P. gingivalis strains were grown anaerobically to early exponential phase in BHI broth at 37°C, H 2 O 2 was added to the cultures, and the cultures further incubated for 24 h. Each

    Article Snippet: P. gingivalis strains were grown in brain heart infusion (BHI) broth supplemented with 0.5% yeast extract (Difco Laboratories, Detroit, MI), hemin (5 μg ml−1 ), vitamin K (0.5 μg ml−1 ), and cysteine (0.1%; Sigma-Aldrich, St. Louis, MO).

    Techniques: Incubation

    Beta-lactam protection in a polymicrobial biofilm. Stationary biofilms of H. influenzae Rd and/or M. catarrhalis were established on chamber slides for 24 h and treated with 100 µg/ml ampicillin or ampicillin with 25 µg/ml clavulanate for an additional 24 h. Biofilms were resuspended in sterile PBS, serially diluted, and plated on sBHI medium plus clarithromycin and BHI medium plates for enumeration of viable H. influenzae Rd and M. catarrhalis bacteria, respectively. Data are represented as means ± SEM. *, P

    Journal: mBio

    Article Title: Indirect Pathogenicity of Haemophilus influenzae and Moraxella catarrhalis in Polymicrobial Otitis Media Occurs via Interspecies Quorum Signaling

    doi: 10.1128/mBio.00102-10

    Figure Lengend Snippet: Beta-lactam protection in a polymicrobial biofilm. Stationary biofilms of H. influenzae Rd and/or M. catarrhalis were established on chamber slides for 24 h and treated with 100 µg/ml ampicillin or ampicillin with 25 µg/ml clavulanate for an additional 24 h. Biofilms were resuspended in sterile PBS, serially diluted, and plated on sBHI medium plus clarithromycin and BHI medium plates for enumeration of viable H. influenzae Rd and M. catarrhalis bacteria, respectively. Data are represented as means ± SEM. *, P

    Article Snippet: M. catarrhalis strains were cultivated in brain heart infusion (BHI) medium (Difco), and H. influenzae strains were cultivated in BHI medium supplemented with hemin (ICN Biochemicals) and NAD (Sigma); this medium is referred to herein as supplemented BHI (sBHI).

    Techniques:

    Polymicrobial infection augments M. catarrhalis persistence in vivo . Chinchillas were infected with 10 3 CFU of H. influenzae or H. influenzae luxS , 10 4 CFU of M. catarrhalis , or a mixture of both species. (A) Middle ear effusion fluids were removed for enumeration of viable H. influenzae and M. catarrhalis bacteria by plating on sBHI medium plus clarithromycin or BHI medium, respectively. (B) Bullae were removed at each time point and homogenized for enumeration of viable H. influenzae and M. catarrhalis bacteria, as described above. Data represent the mean results from four experiments ± SEM. ***, P

    Journal: mBio

    Article Title: Indirect Pathogenicity of Haemophilus influenzae and Moraxella catarrhalis in Polymicrobial Otitis Media Occurs via Interspecies Quorum Signaling

    doi: 10.1128/mBio.00102-10

    Figure Lengend Snippet: Polymicrobial infection augments M. catarrhalis persistence in vivo . Chinchillas were infected with 10 3 CFU of H. influenzae or H. influenzae luxS , 10 4 CFU of M. catarrhalis , or a mixture of both species. (A) Middle ear effusion fluids were removed for enumeration of viable H. influenzae and M. catarrhalis bacteria by plating on sBHI medium plus clarithromycin or BHI medium, respectively. (B) Bullae were removed at each time point and homogenized for enumeration of viable H. influenzae and M. catarrhalis bacteria, as described above. Data represent the mean results from four experiments ± SEM. ***, P

    Article Snippet: M. catarrhalis strains were cultivated in brain heart infusion (BHI) medium (Difco), and H. influenzae strains were cultivated in BHI medium supplemented with hemin (ICN Biochemicals) and NAD (Sigma); this medium is referred to herein as supplemented BHI (sBHI).

    Techniques: Infection, In Vivo

    AI-2 promotes M. catarrhalis biofilm development and antibiotic resistance. (A) M. catarrhalis was cultured in BHI medium or BHI medium supplemented with 0.2 µM synthetic AI-2 (DPD) to determine AI-2 production and depletion, as measured by Vibrio harveyi bioluminescence. H. influenzae luxS was cultured in sBHI medium supplemented with DPD to measure depletion. An uninoculated control of BHI medium with DPD shows the minimal degradation of the AI-2 signal during 6 h of incubation at 37°C. (B) Depletion of DPD by M. catarrhalis biofilms were established for 24 h following incubation with 10 µg/ml tetracycline was measured by bioluminescence over a period of 7 h. (C) M. catarrhalis biofilms were established in the presence or absense of DPD and stained with crystal violet for determination of biofilm biomass at 4, 6, 12, 24, and 48 h. Data represent the mean results from three combined experiments, with three replicate wells per experiment. Error bars represent SEM. (D and E) M. catarrhalis biofilms were established for 24 h in the presence (E) or absence (D) of DPD and stained with a viability kit for CLSM visualization of surface coverage and biofilm thickness. (F and G) Z-series images from panels D and E were compressed to show total viable and nonviable staining of biofilms established in the presence (G) or absence (F) of DPD. (H and I) SEM images of 24-h M. catarrhalis biofilms established with (I) or without (H) DPD. (J) M. catarrhalis biofilms were established for 4 h in the presence or absence of DPD and then treated with 6 µg/ml clarithromycin for 24 h and plated for enumeration of viable bacteria. Data represent the means from three replicates ± SEM. *, P

    Journal: mBio

    Article Title: Indirect Pathogenicity of Haemophilus influenzae and Moraxella catarrhalis in Polymicrobial Otitis Media Occurs via Interspecies Quorum Signaling

    doi: 10.1128/mBio.00102-10

    Figure Lengend Snippet: AI-2 promotes M. catarrhalis biofilm development and antibiotic resistance. (A) M. catarrhalis was cultured in BHI medium or BHI medium supplemented with 0.2 µM synthetic AI-2 (DPD) to determine AI-2 production and depletion, as measured by Vibrio harveyi bioluminescence. H. influenzae luxS was cultured in sBHI medium supplemented with DPD to measure depletion. An uninoculated control of BHI medium with DPD shows the minimal degradation of the AI-2 signal during 6 h of incubation at 37°C. (B) Depletion of DPD by M. catarrhalis biofilms were established for 24 h following incubation with 10 µg/ml tetracycline was measured by bioluminescence over a period of 7 h. (C) M. catarrhalis biofilms were established in the presence or absense of DPD and stained with crystal violet for determination of biofilm biomass at 4, 6, 12, 24, and 48 h. Data represent the mean results from three combined experiments, with three replicate wells per experiment. Error bars represent SEM. (D and E) M. catarrhalis biofilms were established for 24 h in the presence (E) or absence (D) of DPD and stained with a viability kit for CLSM visualization of surface coverage and biofilm thickness. (F and G) Z-series images from panels D and E were compressed to show total viable and nonviable staining of biofilms established in the presence (G) or absence (F) of DPD. (H and I) SEM images of 24-h M. catarrhalis biofilms established with (I) or without (H) DPD. (J) M. catarrhalis biofilms were established for 4 h in the presence or absence of DPD and then treated with 6 µg/ml clarithromycin for 24 h and plated for enumeration of viable bacteria. Data represent the means from three replicates ± SEM. *, P

    Article Snippet: M. catarrhalis strains were cultivated in brain heart infusion (BHI) medium (Difco), and H. influenzae strains were cultivated in BHI medium supplemented with hemin (ICN Biochemicals) and NAD (Sigma); this medium is referred to herein as supplemented BHI (sBHI).

    Techniques: Cell Culture, Incubation, Staining, Confocal Laser Scanning Microscopy

    Polymicrobial biofilm formation protects H. influenzae and M. catarrhalis from antibiotic treatment. Single-species or polymicrobial stationary biofilms were established for 4 h and treated with 60 µg/ml trimethoprim-sulfamethoxazole (TS) (A) or 6 µg/ml clarithromycin (B) for 24 h. Biofilms were resuspended in sterile PBS, serially diluted, and plated on sBHI medium plus clarithromycin or BHI medium for enumeration of viable H. influenzae and M. catarrhalis bacteria, respectively. Data are represented as the mean results from three combined experiments, with two replicates per experiment. Error bars represent SEM. *, P

    Journal: mBio

    Article Title: Indirect Pathogenicity of Haemophilus influenzae and Moraxella catarrhalis in Polymicrobial Otitis Media Occurs via Interspecies Quorum Signaling

    doi: 10.1128/mBio.00102-10

    Figure Lengend Snippet: Polymicrobial biofilm formation protects H. influenzae and M. catarrhalis from antibiotic treatment. Single-species or polymicrobial stationary biofilms were established for 4 h and treated with 60 µg/ml trimethoprim-sulfamethoxazole (TS) (A) or 6 µg/ml clarithromycin (B) for 24 h. Biofilms were resuspended in sterile PBS, serially diluted, and plated on sBHI medium plus clarithromycin or BHI medium for enumeration of viable H. influenzae and M. catarrhalis bacteria, respectively. Data are represented as the mean results from three combined experiments, with two replicates per experiment. Error bars represent SEM. *, P

    Article Snippet: M. catarrhalis strains were cultivated in brain heart infusion (BHI) medium (Difco), and H. influenzae strains were cultivated in BHI medium supplemented with hemin (ICN Biochemicals) and NAD (Sigma); this medium is referred to herein as supplemented BHI (sBHI).

    Techniques:

    pPD1 enhances EF competition for an intestinal niche (a) Bacteriocin assay by the soft agar method with EF r +pPD1::Δ bacAB, bacA-E +, EF r +pPD1 and EF r +pPD1::Δ bacAB spotted on a lawn of susceptible EF. Mice (N=5/group) were given EF r or EF r +pPD1 or EF r +pPD1::Δ bacAB or sterile drinking water for 14 days, at which time all mice were given sterile water. (b) One week and (c) four weeks after withdrawal of EF from drinking water, fecal samples were collected and abundance of total enterococci was determined by enumeration on m-Enterococcus selective agar (Ent agar, ○). Lab strain of EF was enumerated using Ent agar with rifampicin (Rf) (○), or BHI agar with rifampicin (○). (d and e and Fig. 1e ) At the end of week four, animals were euthanized and abundance of EF was determined in distal small intestine (DSI) and large intestine (LI). The results shown are representative of two biologically independent experiments. Horizontal lines indicate geometric mean. Each symbol represents an individual animal.

    Journal: Nature

    Article Title: Bacteriocin production augments niche competition by enterococci in the mammalian GI tract

    doi: 10.1038/nature15524

    Figure Lengend Snippet: pPD1 enhances EF competition for an intestinal niche (a) Bacteriocin assay by the soft agar method with EF r +pPD1::Δ bacAB, bacA-E +, EF r +pPD1 and EF r +pPD1::Δ bacAB spotted on a lawn of susceptible EF. Mice (N=5/group) were given EF r or EF r +pPD1 or EF r +pPD1::Δ bacAB or sterile drinking water for 14 days, at which time all mice were given sterile water. (b) One week and (c) four weeks after withdrawal of EF from drinking water, fecal samples were collected and abundance of total enterococci was determined by enumeration on m-Enterococcus selective agar (Ent agar, ○). Lab strain of EF was enumerated using Ent agar with rifampicin (Rf) (○), or BHI agar with rifampicin (○). (d and e and Fig. 1e ) At the end of week four, animals were euthanized and abundance of EF was determined in distal small intestine (DSI) and large intestine (LI). The results shown are representative of two biologically independent experiments. Horizontal lines indicate geometric mean. Each symbol represents an individual animal.

    Article Snippet: Brain heart infusion medium (BHI) and m-Enterococcus agar (Difco™) (Ent-agar) were prepared as described by the manufacturer (Becton Dickinson).

    Techniques: Mouse Assay

    EF+pPD1 but not EF r +pCF10 dominates the intestinal enterococcal population pCF10 is a well-studied pheromone-inducible conjugative plasmid of EF that encodes resistance to tetracycline but does not encode a known bacteriocin determinant. (a) EF r : (o, N=3 mice, no plasmid), EF r +pPD1: (☐, N=3 mice, pPD1) or EF r +pCF10, (∇, N=5 mice, pCF10) was added to drinking water for 14 days, and then replaced by sterile drinking water. (a) Fecal samples were taken from each animal at the transition to sterile drinking water (week 0) and then weekly. EF abundance was determined by enumeration on BHI agar with rifampicin. (b) Four weeks after withdrawal of EF from drinking water, animals were euthanized and abundance of EF r +pCF10 was determined in each segment of the GI tract (distal small intestine, DSI; cecum; and large intestine, LI). Mice colonized with EF r +pCF10 maintained long-term fecal shedding of EF r +pCF10 similar to EF r , and persistent colonization throughout the GI tract. Abundance of enterococci in the feces was determined by enumeration on m-Enterococcus agar (Ent agar, o), Ent agar with rifampicin (Rf) (○), or BHI agar with rifampicin (○) at week 1 (c) and week 4 (d). Unlike EF r +pPD1 that dominated the enterococcal niche in the GI tract ( Fig. 1e ), EF r +pCF10 did not outcompete the indigenous enterococci, colonizing at levels comparable to EF r (c and d). (a) The lines are fitted using an exponential decay model, and there are significant differences in rate of decay for the “pCF10” group vs. “pPD1” group ( P =0.012) and for the “pPD1” vs. “no plasmid” group ( P =0.007). Horizontal lines in (b, c, and d) indicate geometric mean. Each symbol represents an individual animal; data are obtained from one experiment.

    Journal: Nature

    Article Title: Bacteriocin production augments niche competition by enterococci in the mammalian GI tract

    doi: 10.1038/nature15524

    Figure Lengend Snippet: EF+pPD1 but not EF r +pCF10 dominates the intestinal enterococcal population pCF10 is a well-studied pheromone-inducible conjugative plasmid of EF that encodes resistance to tetracycline but does not encode a known bacteriocin determinant. (a) EF r : (o, N=3 mice, no plasmid), EF r +pPD1: (☐, N=3 mice, pPD1) or EF r +pCF10, (∇, N=5 mice, pCF10) was added to drinking water for 14 days, and then replaced by sterile drinking water. (a) Fecal samples were taken from each animal at the transition to sterile drinking water (week 0) and then weekly. EF abundance was determined by enumeration on BHI agar with rifampicin. (b) Four weeks after withdrawal of EF from drinking water, animals were euthanized and abundance of EF r +pCF10 was determined in each segment of the GI tract (distal small intestine, DSI; cecum; and large intestine, LI). Mice colonized with EF r +pCF10 maintained long-term fecal shedding of EF r +pCF10 similar to EF r , and persistent colonization throughout the GI tract. Abundance of enterococci in the feces was determined by enumeration on m-Enterococcus agar (Ent agar, o), Ent agar with rifampicin (Rf) (○), or BHI agar with rifampicin (○) at week 1 (c) and week 4 (d). Unlike EF r +pPD1 that dominated the enterococcal niche in the GI tract ( Fig. 1e ), EF r +pCF10 did not outcompete the indigenous enterococci, colonizing at levels comparable to EF r (c and d). (a) The lines are fitted using an exponential decay model, and there are significant differences in rate of decay for the “pCF10” group vs. “pPD1” group ( P =0.012) and for the “pPD1” vs. “no plasmid” group ( P =0.007). Horizontal lines in (b, c, and d) indicate geometric mean. Each symbol represents an individual animal; data are obtained from one experiment.

    Article Snippet: Brain heart infusion medium (BHI) and m-Enterococcus agar (Difco™) (Ent-agar) were prepared as described by the manufacturer (Becton Dickinson).

    Techniques: Plasmid Preparation, Mouse Assay

    Complementation of Bac-21 production restores colonization phenotype by providing competitive advantage Bacteriocin activity was restored upon ectopic expression of bacABCDE (from pAM401) in EF+pPD1::Δ bacAB but not in EF lacking pPD1, indicating that the distal part of the bac operon ( bacFGHI) is necessary for bacteriocin expression and that the bacAB in-frame deletion is not polar on downstream genes. Mice (N=5) were given EF r +pPD1::Δ bacAB, bacABCDE + as described in the methods and abundance was determined by enumeration on m-Enterococcus agar (Ent agar), Ent agar plus rifampicin (Rf), or BHI agar with rifampicin. The presence of pAM401A:: bacABCDE+ (complementing plasmid) was determined by enumerating CFU on BHI agar with rifampicin and chloramphenicol (Cm). Fecal samples were obtained at week 1 (a) and week 4 (b) after transition to sterile drinking water. Horizontal lines indicate geometric mean. Each symbol represents an individual animal and data is a representative of two biologically independent experiments. EF s +pPD1:: ΔbacAB bacABCDE + stably colonized the GI tract (a), although in the absence of chloramphenicol selection, pAM401:: bacABCDE was gradually lost from the population (b). Over time, loss of pAM401:: bacABCDE resulted in the complemented strain reverting back to the bacteriocin-defective Δ bacAB strain, with the loss of bacteriocin activity. Nevertheless, this strain persisted in the gut, suggesting that Bac-21 was essential for clearing a niche for EF, but once cleared, EF uses other mechanisms to maintain colonization (a and b).

    Journal: Nature

    Article Title: Bacteriocin production augments niche competition by enterococci in the mammalian GI tract

    doi: 10.1038/nature15524

    Figure Lengend Snippet: Complementation of Bac-21 production restores colonization phenotype by providing competitive advantage Bacteriocin activity was restored upon ectopic expression of bacABCDE (from pAM401) in EF+pPD1::Δ bacAB but not in EF lacking pPD1, indicating that the distal part of the bac operon ( bacFGHI) is necessary for bacteriocin expression and that the bacAB in-frame deletion is not polar on downstream genes. Mice (N=5) were given EF r +pPD1::Δ bacAB, bacABCDE + as described in the methods and abundance was determined by enumeration on m-Enterococcus agar (Ent agar), Ent agar plus rifampicin (Rf), or BHI agar with rifampicin. The presence of pAM401A:: bacABCDE+ (complementing plasmid) was determined by enumerating CFU on BHI agar with rifampicin and chloramphenicol (Cm). Fecal samples were obtained at week 1 (a) and week 4 (b) after transition to sterile drinking water. Horizontal lines indicate geometric mean. Each symbol represents an individual animal and data is a representative of two biologically independent experiments. EF s +pPD1:: ΔbacAB bacABCDE + stably colonized the GI tract (a), although in the absence of chloramphenicol selection, pAM401:: bacABCDE was gradually lost from the population (b). Over time, loss of pAM401:: bacABCDE resulted in the complemented strain reverting back to the bacteriocin-defective Δ bacAB strain, with the loss of bacteriocin activity. Nevertheless, this strain persisted in the gut, suggesting that Bac-21 was essential for clearing a niche for EF, but once cleared, EF uses other mechanisms to maintain colonization (a and b).

    Article Snippet: Brain heart infusion medium (BHI) and m-Enterococcus agar (Difco™) (Ent-agar) were prepared as described by the manufacturer (Becton Dickinson).

    Techniques: BAC Assay, Activity Assay, Expressing, Mouse Assay, Plasmid Preparation, Stable Transfection, Selection