s aureus newman strain  (ATCC)

Journal: Antimicrobial Agents and Chemotherapy

Article Title: Genome-wide screen reveals glycerol-induced aminoglycoside potentiation against Staphylococcus aureus via boosting GlpK-initiated energy metabolism

doi: 10.1128/aac.00938-25

Figure Lengend Snippet: GlpK is essential for glycerol-enhanced tobramycin uptake and lethality via boosting glycerol-initiated energy metabolism. ( A ) Inhibition of E. coli cell growth on LB agar dishes by tobramycin extracted from stationary-phase S. aureus ATCC 25923 wild-type and ARTP-3A mutant cells, which were pretreated with 100 µg/mL tobramycin dissolved in pure water or 0.3 M glycerol. ( B ) Survival of S. aureus Newman wild-type and Δ glpK mutant cells following the combined treatment with 50 µg/mL tobramycin plus 0.3 M glycerol. ( C ) Inhibition of E. coli cell growth on LB agar dishes by tobramycin extracted from stationary-phase S. aureus Newman wild-type and Δ glpK mutant cells, which were pretreated with 100 µg/mL tobramycin dissolved in pure water or 0.3 M glycerol. ( D, E ) Results of a flow cytometric analysis of stationary-phase S. aureus ARTP-3A mutant cells (panel D ) as well as Newman wild-type and Δ glpK mutant cells (panel E) after treatment with indicated buffers (phosphate-buffered saline [PBS] or glycerol), similarly to . ( F, G ) ATP levels of indicated S. aureus cells after glycerol treatment. ( H ) Growth curves of Newman wild-type and Δ glpK mutant cells as cultured in 10% LB medium plus 0.3 M glycerol. ( I ) 3-D structure of GlpK (PDB ID: 3g25). Left: tetrameric structure of GlpK; right part: monomeric structure of GlpK showing its glycerol- and ATP-binding sites (colored in green and orange, respectively) and missense variation sites from our tolerant clones (colored in red). ( J ) Overlapping of missense variation sites of GlpK (colored in red on the top) with its ATP- and glycerol-binding sites (colored in green and orange on the bottom, respectively).

Article Snippet: Second, we constructed a glpK deletion mutant in the S. aureus Newman strain (Note: gene deletion of glpK was not successfully achieved in the ATCC 25923 strain due to the failure of plasmid transformation into this strain by electroporation), and found that the Δ glpK mutant was fully tolerant to the tobramycin-glycerol combined treatment, whereas the Newman wild-type strain could be killed by around two orders of magnitude ( ).

Techniques: Inhibition, Mutagenesis, Saline, Cell Culture, Binding Assay, Clone Assay

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Isoliquiritigenin Attenuates Staphylococcus aureus Adhesion and Invasion to Counteract Staphylococcus aureus Pathogenicity and Infection

doi: 10.3389/fcimb.2025.1686699

Figure Lengend Snippet: Inhibition of S. aureus adhesion, invasion, and biofilm formation by ISL through SrtA suppression. (A, B) ISL inhibits S. aureus adhesion to fibrinogen in a dose-dependent manner. Treatment with 32 μg/ml ISL reduced the adhesion of S. aureus USA300 and Newman. (C) ISL significantly reduces the invasive capacity of S. aureus USA300 against A549 lung epithelial cells in a concentration-dependent manner, with 32 μg/ml ISL resulting in a marked reduction in bacterial invasion. (D, E) ISL inhibits biofilm formation in both S. aureus USA300 and Newman strains. (F) ISL primarily disrupts the early stages of biofilm formation, with no significant effect on mature biofilms. (G) ISL reduces the surface expression of SpA, as evidenced by flow cytometry analysis, indicating that SpA anchoring is suppressed through the inhibition of SrtA activity. (H) Western blot analysis revealed that ISL does not alter SrtA expression. n.s., not significant (P ≥ 0.05); *, P < 0.05; **, P < 0.01; ***, P < 0.001.

Article Snippet: The methicillin-resistant Staphylococcus aureus (MRSA) USA300 strain and methicillin-sensitive S. aureus (MSSA) Newman strain utilized throughout this study were sourced from the American Type Culture Collection (ATCC; Manassas, VA, United States).

Techniques: Inhibition, Concentration Assay, Expressing, Flow Cytometry, Activity Assay, Western Blot

Journal: Scientific Reports

Article Title: Assessment of the biofilm formation capacities of Staphylococcus aureus strains Newman and Newman D2C in vitro and in vivo

doi: 10.1038/s41598-025-00521-5

Figure Lengend Snippet: Whole-genome sequencing reveals disparities between the Newman/Newman D2C derivatives in stock at the Institute for Medical Microbiology and Hygiene (Homburg, Germany) and published Newman/Newman D2C reference genomes. A visualization of S. aureus Newman/Newman D2C genetic diversity is shown on a circular map of the chromosome of Newman UoM ( LT598688.1 ). The first ring from the outside shows the scale of the chromosome in kilobases (kb). The second ring indicates the positions of mutated genes in the genome (gray). The next five circles illustrate the five genomes used in this study (Newman D2C HOM [ CP160002.1 ], Newman D2C [ CP023391.1 ], Newman [ AP009351.1 ], Newman HOM [ CP160003.1 ], Newman UoM [ NZ_LT598688.1 ]). The colored tiles inside each circle represent the positions of mutations within each respective genome.

Article Snippet: Newman D2C HOM , Clumping factor-positive variant of S. aureus strain Newman D2 (ATCC 25904), agrA , saeR , .

Techniques: Sequencing

Journal: Scientific Reports

Article Title: Assessment of the biofilm formation capacities of Staphylococcus aureus strains Newman and Newman D2C in vitro and in vivo

doi: 10.1038/s41598-025-00521-5

Figure Lengend Snippet: Quantitative transcript analyses of RNAIII and eap by qRT-PCR in Newman HOM (orange bars) and Newman D2C HOM (turquoise bars) cells grown in TSB at 37 °C and 225 rpm. ( a , b ) Transcript rates of RNAIII ( a ) and eap ( b ) at the time points indicated. Transcript rates were quantified in reference to the transcription of gyrase B (in copies per copy of gyrB ). Data are presented as mean + SD of six biological replicates. ** p < 0.01 (Mann–Whitney U test between Newman HOM and Newman D2C HOM at a given time point).

Article Snippet: Newman D2C HOM , Clumping factor-positive variant of S. aureus strain Newman D2 (ATCC 25904), agrA , saeR , .

Techniques: Quantitative RT-PCR, MANN-WHITNEY

Journal: Scientific Reports

Article Title: Assessment of the biofilm formation capacities of Staphylococcus aureus strains Newman and Newman D2C in vitro and in vivo

doi: 10.1038/s41598-025-00521-5

Figure Lengend Snippet: Growth kinetics of strains Newman HOM and Newman D2C HOM under aerobic conditions in tryptic soy broth (TSB). Bacteria were inoculated to an optical density at 600 nm (OD 600 ) of 0.05 in TSB and cultured aerobically at 37 °C and 225 rpm in a culture-to-flask volume of 1:10. OD 600 measurements of the cultures were determined hourly. Samples were diluted in TSB when an OD 600 value of 0.8 was reached. The dilution factor was used to multiply with the measured result for the OD 600 values displayed in the graphs. Calculated values are given as arbitrary units (au). ( a ) Growth kinetics of Newman HOM (orange symbols) and Newman D2C HOM (turquoise symbols) cell suspensions. The results are the mean ± SD of nine biological replicates. ( b , c ) OD 600 values ( b ) and colony forming units (CFU) rates per ml ( c ) of the cell cultures at 2, 4, 6, and 8 h of growth, respectively. The data represent the values of every individual OD 600 reading/CFU count (symbols) and the median (horizontal line). ( d ) Fold changes in the CFU rates per ml of Newman HOM and Newman D2C HOM 6 h cultures upon sonication. Data are presented as box and whisker plots (min-to-max). Symbols indicate the mean values per experiment ( n = 6). ( e ) Forward scatter (FSC) versus green fluorescence cytograms of SYTO9-stained Newman HOM and Newman D2C HOM 6 h cultures (diluted 1:100 in PBS). Data shown represent one of the assays carried out in three biological replicates. sc single cells, d doublets, la larger aggregates. ** p < 0.01 [Mann–Whitney U test ( b , d ) and Kolmogorov–Smirnov test with two-stage linear step-up procedure of Benjamini, Krieger, and Yekutieli ( c )].

Article Snippet: Newman D2C HOM , Clumping factor-positive variant of S. aureus strain Newman D2 (ATCC 25904), agrA , saeR , .

Techniques: Bacteria, Cell Culture, Sonication, Whisker Assay, Fluorescence, Staining, MANN-WHITNEY

Journal: Scientific Reports

Article Title: Assessment of the biofilm formation capacities of Staphylococcus aureus strains Newman and Newman D2C in vitro and in vivo

doi: 10.1038/s41598-025-00521-5

Figure Lengend Snippet: Quantitative and qualitative adhesion capacities of strains Newman HOM and Newman D2C HOM on artificial surfaces. ( a ) Quantitative adhesion of cells of strains Newman HOM (orange filled box and symbols) and Newman D2C HOM (turquoise filled box and symbols) on polystyrene-based 96-well microplates. Exponential growth phase cells were fluorescence-labeled with Hoechst 33342 and placed into the wells of the Nunclon Delta-treated, flat-bottom 96-well microplate. Unbound and loosely bound bacterial cells were removed by washing, and adherent cells were counted by fluorescence microscopy. Box and whisker plots (min-to-max) of the mean values of adherent bacteria found in a 100 µm 2 area ( n = 9 images/strain obtained by three biological experiments). ( b–e ) Adhesive strengths of cells of strains Newman HOM and Newman D2C HOM on polystyrene and PVC tubing. ( b , c ) Mean SCFS retraction curves of six individual bacteria (indicated by different colors) per strain on polystyrene ( b ) and PVC tubing ( c ), respectively. Bacterial cells were probed with the substratum with a 5 s surface delay time. The means were calculated using 30 force-distance curves per cell and surface type (shaded areas represent the standard deviations per bacterial cell). ( d , e ) Box and whisker plots (min-to-max) of the mean adhesion forces ( d ) and mean rupture lengths ( e ) of cells of strains Newman HOM and Newman D2C HOM probed on polystyrene and PVC tubing, respectively. Mean values of the individual cells are indicated as round symbols. ns not significant; ** p < 0.01 (Mann–Whitney U test).

Article Snippet: Newman D2C HOM , Clumping factor-positive variant of S. aureus strain Newman D2 (ATCC 25904), agrA , saeR , .

Techniques: Fluorescence, Labeling, Microscopy, Whisker Assay, Bacteria, Adhesive, MANN-WHITNEY

Journal: Scientific Reports

Article Title: Assessment of the biofilm formation capacities of Staphylococcus aureus strains Newman and Newman D2C in vitro and in vivo

doi: 10.1038/s41598-025-00521-5

Figure Lengend Snippet: Biofilm formation of S. aureus strains SA113, Newman HOM, and Newman D2C HOM in TSB supplemented with 0.5% glucose. Exponential growth phase cells of strains SA113 (red bars), Newman HOM (orange bars), and Newman D2C HOM (turquoise bars) were inoculated into TSB supplemented with 0.5% glucose (TSB-G) and cultured in Nunclon Delta-treated 96-well microplates for 18 h at 37 °C under static or dynamic (120 rpm) conditions as indicated. ( a–d ) Vegetation were washed twice prior to the safranin staining ( a , b ) or washed as indicated ( c , d ). ( a , c ) Representative images of safranin-stained vegetation. ( b , d ) A 530 readings of safranin contents in the wells after solubilization with 30% acetic acid. ( e ) Impact of the washing steps on the A 530 readings of safranin contents in the wells. Values are given in relation to the safranin signals seen in wells that were not washed, which were set to 100%. ( f ) PIA contents of the vegetation formed by strains SA113, Newman HOM, and Newman D2C HOM in TSB-G in 96-well microplates cultured for 18 h at 37 °C under static conditions. PIA contents of the biofilms were determined by staining the vegetation with XFD488-labeled WGA (2.5 µg/ml) and determining the fluorescence signals of the incorporated dye at 480/530 nm (Excitation/Emission). Differences in PIA contents are given as relative light units (RFUs). Results represent the averages of five to nine independent experiments done in duplicate. Error bars indicate the standard deviation of the mean. Round symbols indicate the mean A 530 /%/RFU values of individual experiments. * p < 0.05; ** p < 0.01 (Kruskal–Wallis test and Dunn’s multiple comparison test).

Article Snippet: Newman D2C HOM , Clumping factor-positive variant of S. aureus strain Newman D2 (ATCC 25904), agrA , saeR , .

Techniques: Cell Culture, Staining, Labeling, Fluorescence, Standard Deviation, Comparison

Journal: Scientific Reports

Article Title: Assessment of the biofilm formation capacities of Staphylococcus aureus strains Newman and Newman D2C in vitro and in vivo

doi: 10.1038/s41598-025-00521-5

Figure Lengend Snippet: Three-dimensional organization and PIA distribution of the vegetation formed by strains SA113, Newman HOM, and Newman D2C HOM in PS-based microplates. Exponential growth phase cells of S. aureus strains were inoculated into TSB supplemented with 0.5% glucose and cultured in tissue culture-treated, PS-based 6-well microplates for 18 h at 37 °C under static conditions. The vegetation formed were stained with XFD488-WGA and Nile Red (NR), and fluorescence was monitored with CLSM. ( a , b ) Three-dimensional image reconstructions of z series were recorded at 525 nm (XFD488) and 595 nm (NR), respectively. Reconstructed top view ( a ) and side view ( b ) images of the vegetation formed by the test strains. CLSM reconstructions are representative of three separate experiments. Each side of a grid square in the image reconstructions represents 100 μm. Scale bar, 200 μm (scale bar applies to all images in the respective panel).

Article Snippet: Newman D2C HOM , Clumping factor-positive variant of S. aureus strain Newman D2 (ATCC 25904), agrA , saeR , .

Techniques: Cell Culture, Staining, Fluorescence

Journal: Scientific Reports

Article Title: Assessment of the biofilm formation capacities of Staphylococcus aureus strains Newman and Newman D2C in vitro and in vivo

doi: 10.1038/s41598-025-00521-5

Figure Lengend Snippet: Biofilm formation of S. aureus strains SA113, Newman HOM, and Newman D2C HOM in TSB-HBS or RPMI-1640 under static conditions. ( a , b ) Exponential growth phase cells of strains SA113 (red bars), Newman HOM (orange bars), and Newman D2C HOM (turquoise bars) were inoculated into TSB supplemented with 5% human blood serum (TSB-HBS; a ) or Roswell Park Memorial Institute 1640 medium (RPMI-1640; b ) and cultured in Nunclon Delta-treated 96-well microplates for 18 h at 37 °C under static conditions. Washing steps prior to the safranin staining were performed as indicated. ( a , c ) Representative images of safranin-stained vegetation. ( b , d ) A 530 readings of safranin contents in the wells after solubilization with 30% acetic acid. Results represent the averages of six to nine independent experiments done in duplicate. Error bars indicate the standard deviation of the mean. Round symbols indicate the mean A 530 values of individual experiments. * p < 0.05; ** p < 0.01 (Kruskal–Wallis test and Dunn’s multiple comparison test).

Article Snippet: Newman D2C HOM , Clumping factor-positive variant of S. aureus strain Newman D2 (ATCC 25904), agrA , saeR , .

Techniques: Cell Culture, Staining, Standard Deviation, Comparison

Journal: Scientific Reports

Article Title: Assessment of the biofilm formation capacities of Staphylococcus aureus strains Newman and Newman D2C in vitro and in vivo

doi: 10.1038/s41598-025-00521-5

Figure Lengend Snippet: Biofilm formation of S. aureus strains SA113, Newman HOM, and Newman D2C HOM on PVC tubing under dynamic conditions. ( a ) Cell suspensions of strains SA113 (red bars), Newman HOM (orange bars), and Newman D2C HOM (turquoise bars) were used to inoculate the lumen of 1 cm long PU-based PVC tubing fragments, and the infected tubing fragments were cultured in TSB or RPMI-1640 for 5 days at 37 °C under non-nutrient limiting and dynamic conditions (rotation at 20 rpm). ( b ) Biofilm formation of S. aureus strains SA113, Newman HOM, and Newman D2C HOM on the lumen of PVC tubing in TSB under constant flow (1 ml/min). CFU rates of detached biofilms are shown. Error bars indicate the standard deviation of the mean. Round symbols indicate the CFU values of individual experiments. ns not significant; * p < 0.05 (Kruskal–Wallis test and Dunn’s multiple comparison test).

Article Snippet: Newman D2C HOM , Clumping factor-positive variant of S. aureus strain Newman D2 (ATCC 25904), agrA , saeR , .

Techniques: Infection, Cell Culture, Standard Deviation, Comparison

Journal: Scientific Reports

Article Title: Assessment of the biofilm formation capacities of Staphylococcus aureus strains Newman and Newman D2C in vitro and in vivo

doi: 10.1038/s41598-025-00521-5

Figure Lengend Snippet: Infectivity of S. aureus strains Newman HOM and Newman D2C HOM in a murine foreign body-related infection model. Sterile polyurethane-based peripheral venous catheter tubing fragments were implanted subcutaneously into the left and right flanks of normoglycemic C57BL/6 mice ( n = 5 per group) and inoculated with cells of S. aureus strain Newman HOM (orange symbols) and Newman D2C HOM (turquoise symbols), respectively. Mice with uninfected implants served as controls (white symbols). On day 10 after infection, animals were euthanized, edema sizes around the insertion site were determined, blood was taken, and the catheter fragments and the surrounding tissues (peri-implant tissues) were removed and separated. Bacteria adherent to the catheters were detached by sonication in saline, and tissue samples were homogenized in saline. Bacterial loads from catheter-detached biofilms ( a ) and in peri-implant tissue homogenates ( b ) were determined by CFU counting. Edema endpoints at day 10 post-infection are depicted in ( c ), and neutrophil numbers observed in the blood are depicted in ( d ). Each symbol represents an individual infection site, and horizontal bars indicate the median of all observations. ns, not significant; * p < 0.05; ** p < 0.01 (Kruskal–Wallis test and Dunn’s multiple comparison test).

Article Snippet: Newman D2C HOM , Clumping factor-positive variant of S. aureus strain Newman D2 (ATCC 25904), agrA , saeR , .

Techniques: Infection, Sterility, Bacteria, Sonication, Saline, Comparison

Journal: Microbiology Spectrum

Article Title: A mechanistic understanding of the effect of Staphylococcus aureus VraS histidine kinase single-point mutation on antibiotic resistance

doi: 10.1128/spectrum.00095-25

Figure Lengend Snippet: Relationship between the drug concentrations and the bacterial burden. Dose-response curves of the tested antibiotics in the Newman WT and T331I mutant strain. Results obtained are in , and the parameters for the model are in Table S1.

Article Snippet: The drug-susceptible strain S. aureus Newman D2C strain (ATCC #25904) and its isogenic mutant VraS T331I were maintained in Luria-Bertani (LB) media at 37°C unless otherwise mentioned.

Techniques: Mutagenesis