S aureus atcc 43300

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S. aureus ATCC 43300 is a bacterial strain from the American Type Culture Collection. It is a methicillin-resistant Staphylococcus aureus (MRSA) strain.

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S. aureus (168 citations) ATCC 43300 (34 citations) Methicillin-resistant S. aureus ATCC 43300 (3 citations)

21 protocols using s aureus atcc 43300

Characterization of MRSA and MSSA Strains

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S. aureus ATCC 43300 (methicillin resistant S.aureus [MRSA]) and S.aureus ATCC 29213 (methicillin sensitive S.aureus [MSSA]) from ATCC, Mannasse, USA were used in this study. Clinical isolates of S. aureus were procured from Post-graduate Institute of Medical Education and Research (PGIMER), Chandigarh, India. The strains were isolated from clinical specimens (nasal screening swabs, blood, pus, soft tissue, wound swabs, respiratory samples and body fluids) from both in-patient as well as out-patients from in as well as around places near Chandigarh, India.
These strains were identified on the basis of Gram reaction, growth on mannitol salt agar (MSA), catalase activity, and coagulase test. Methicillin resistance was determined using cefoxitin disk on Mueller-Hinton agar (Oxoid) followed by determination of MICs of oxacillin for these strains as recommended by Clinical and Laboratory Standards Institute (CLSI) [38] .A total of forty five MRSA isolates were selected and stored in glycerol at −80°C. These strains were used for determining the lytic spectrum/host range of the isolated phage. S. aureus specific lytic bacteriophage, MR-5 with broad host range (active against both MRSA as well as MSSA strains), belonging to family Myoviridae, isolated and characterized in our laboratory earlier was used in the present study [39] (link).

Kaur S., Harjai K, & Chhibber S. (2014). Bacteriophage Mediated Killing of Staphylococcus aureus In Vitro on Orthopaedic K Wires in Presence of Linezolid Prevents Implant Colonization. PLoS ONE, 9(3), e90411.

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Survival Assay of C. elegans Infected with MRSA Strains

Cited 1 time

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The C. elegans mutant strain SS104
was purchased from the Caenorhabditis Genetics Centre and was propagated
and synchronized as previously reported.^{57 (link)} Approximately 10–20 worms were added per well of a 96-well
plate. Each control and experimental treatment had six replicate wells
(n ∼60–120 worms per treatment). E. coli OP50 culture was added as an uninfected control. S. aureus ATCC 43300, USA100, USA300, and COL were
purchased from ATCC. Cultures were grown in tryptic soy broth (TSB)
at 37 °C shaking overnight. They were normalized to an OD₆₀₀ of 1.2 and added to the worms as infected treatments. Loratadine
was used at a final concentration of 50 μM and oxacillin was
used at a final concentration of 4 μg mL^–1 to be consistent with RNA-seq and RT-qPCR experiments. The plate
was kept at room temperature on a rocking platform for 7 days. Dead
worms were scored every 24 h. Kaplan–Meier survival curves
and median survival times were generated using GraphPad Prism. Statistically
significant differences (p < 0.05) between survival
curves were determined using the log-rank (Mantel–Cox) test
within Prism. Four independent experiments were conducted for each
strain of MRSA, with a representative experiment shown.

Viering B.L., Balogh H., Cox C.F., Kirpekar O.K., Akers A.L., Federico V.A., Valenzano G.Z., Stempel R., Pickett H.L., Lundin P.M., Blackledge M.S, & Miller H.B. (2023). Loratadine Combats Methicillin-Resistant Staphylococcus aureus by Modulating Virulence, Antibiotic Resistance, and Biofilm Genes. ACS Infectious Diseases, 10(1), 232-250.

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Staphylococcus aureus Culture Protocol

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In this study, gram-positive Staphylococcus aureus (S. aureus) (ATCC 43300) was obtained from the American Type Culture Collection (Rockefeller, MD) in freeze-dried form. The bacteria were cultured on sheep blood agar (SBA) plates at 37 °C overnight. Then, a single bacterial colony was carefully transferred to 3 mL of TSB and incubated on an orbital shaker for another 12 h. The purified bacterial solution was gradient diluted to a suitable concentration for in vitro and in vivo experiments.

Huang M., Ye K., Hu T., Liu K., You M., Wang L, & Qin H. (2021). Silver Nanoparticles Attenuate the Antimicrobial Activity of the Innate Immune System by Inhibiting Neutrophil-Mediated Phagocytosis and Reactive Oxygen Species Production. International Journal of Nanomedicine, 16, 1345-1360.

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Cell and Bacterial Cell Culture Protocol

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MC3T3-E1 osteoblast precursor cells and RAW264.7 macrophages were purchased from Shanghai Institute of Cell Biology, Chinese Academy of Science. S. aureus (ATCC43300) and E. coli (ATCC35218) were obtained from the American Type Culture Collection. The cells were cultured in Dulbecco’s modified Eagle’s medium (Gibco) containing 10% fetal bovine serum (Gibco). Bacteria were grown in trypticase soy broth (TSB, Solarbio, China). The cells and bacteria were passaged every 1 to 2 days.

Yang C., Luo Y., Shen H., Ge M., Tang J., Wang Q., Lin H., Shi J, & Zhang X. (2022). Inorganic nanosheets facilitate humoral immunity against medical implant infections by modulating immune co-stimulatory pathways. Nature Communications, 13, 4866.

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Microbial Strain Collection and Characteristics

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Staphylococcus aureus ATCC 29213, Staphylococcus aureus ATCC 25923, S. aureus ATCC 43300, Enterococcus faecalis ATCC 29212, Enterococcus faecium ATCC 70022, Escherichia coli ATCC 25922, and Pseudomonas aeruginosa ATCC 27853 have been obtained from the American Type Culture Collection (ATCC) via Microbiologics Inc. (St. cloud, MN), E. coli MG1655 originated from the laboratory collection of Dr. Csaba Pál. E. coli BW 25113, and E. coli BW 25113 ΔacrB, ΔdapF, ΔmrcB and ΔsurA mutant strains originated from the KEIO collection [30 ]. Single-gene knock-out strains of E. coli, JW5503 ΔtolC [30 ] and JD17464 ΔlpxC were obtained from the NBRP-E.coli collection at the National Institute of Genetics (NIG, Japan).

Durcik M., Lovison D., Skok Ž., Durante Cruz C., Tammela P., Tomašič T., Benedetto Tiz D., Draskovits G., Nyerges Á., Pál C., Ilaš J., Peterlin Mašič L., Kikelj D, & Zidar N. (2018). New N-phenylpyrrolamide DNA gyrase B inhibitors: Optimization of efficacy and antibacterial activity. European journal of medicinal chemistry, 154, 117-132.

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Cultivation and Preparation of S. aureus Strains

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S. aureus ATCC 29213 and S. aureus ATCC 43300 were obtained from the American Type Culture Collection (Rockville, MD, USA). All the strains were cultured in tryptic soya broth (TSB, OXOID) under aerobic conditions at 37 °C for 24 h on an orbital shaker at 200 rpm. Each tested compound was dissolved in methanol (1 mg/200 μL) to give a stock solution and diluted in Mueller–Hinton (MH).

Guzzo F., Buommino E., Landrum L., Russo R., Lembo F., Fiorentino A, & D’Abrosca B. (2023). Phytochemical Investigation of Myrcianthes cisplatensis: Structural Characterization of New p-Coumaroyl Alkylphloroglucinols and Antimicrobial Evaluation against Staphylococcus aureus. Plants, 12(5), 1046.

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Bacterial Strains and Cell Lines

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The bacterial strains S. aureus ATCC 43300 and S. aureus ATCC 25923 were purchased from American Type Culture Collection (ATCC). S. hyicus NCTC 10350 was purchased from National Collection of Type Culture (NCTC). The clinical strain of S. hyicus ACCC 61734 (Agricultural Culture Collection of China) was obtained from Animal husbandry and veterinary research institute (Tianjin, China). Hacat cells were purchased from Peking Union Medical College (Beijing, China). The six-week-old female ICR mice (SPF) were purchased from the Vital River Laboratories (VRL, Beijing, China). All other chemical reagents used were of analytical grade.

Liu H., Yang N., Teng D., Mao R., Hao Y., Ma X, & Wang J. (2021). Design and Pharmacodynamics of Recombinant Fungus Defensin NZL with Improved Activity against Staphylococcus hyicus In Vitro and In Vivo. International Journal of Molecular Sciences, 22(11), 5435.

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Bacterial Strain Preparation and Preservation

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Escherichia coli ATCC 35218, Bacillus subtilis ATCC 6633, S. aureus ATCC 6538P (methicillin susceptible S. aureus, MSSA), S. aureus ATCC 43300 (MRSA), Enterococcus faecalis ATCC 29212, and E. faecalis ATCC 51299 (VanB phenotype) were obtained from the American Type Culture Collection (ATCC). E. faecalis 9160188401-EF-34 (VanA phenotype) is a clinical isolate, which was kindly provided by Laboratorio Microbiologia Clinica – Ospedale di Circolo, Varese, Italy. E. coli and B. subtilis were propagated overnight in Luria Bertani medium (LB, 2% tryptone, 2% yeast extract, and 1% NaCl), and the S. aureus and E. faecalis strains in Müller Hinton broth 2 (MHB2, 0.3% beef infusion solids, 1.75% casein hydrolysate, and 0.15% starch) with continuous shaking at 200 rpm and 37°C. For exponential growth, overnight cultures were transferred to fresh medium: inocula were prepared to start the cultures with an optical density at 600 nm (OD_{600 nm}) of 0.1 in the final medium. For long-term preservation, bacterial cultures were stored at -20°C in 20% glycerol. Media were acquired from Sigma-Aldrich, Milan, Italy, unless otherwise stated.

Armenia I., Marcone G.L., Berini F., Orlandi V.T., Pirrone C., Martegani E., Gornati R., Bernardini G, & Marinelli F. (2018). Magnetic Nanoconjugated Teicoplanin: A Novel Tool for Bacterial Infection Site Targeting. Frontiers in Microbiology, 9, 2270.

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Bovine Mastitis Staphylococcus aureus Protocols

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NZX (> 90% purity) was prepared as previous protocols (Liu et al. 2020) . Lincomycin and ceftiofur were purchased from the Dalian Meilun Biotech (Dalian, China). S. aureus E48 (CGMCC 1.90006) isolated from bovine mastitis was provided by Professor Xin Zhao, Northwest A&F University (Yangling, China) and stored at the China General Microbiological Culture Collection Center (CGMCC). S. aureus ATCC 43300 and S. aureus ATCC 25923 were purchased from the American Type Culture Collection (ATCC).

Zheng X., Yang N., Mao R., Hao Y., Teng D, & Wang J. (2024). Pharmacokinetics and pharmacodynamics of antibacterial peptide NZX in Staphylococcus aureus mastitis mouse model. Applied microbiology and biotechnology, 108(1).

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Antimicrobial Activity Assessment

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S. aureus ATCC 29213, S. aureus ATCC 43300, S. epidermidis ATCC 12228 and S. epidermidis ATCC 35984, were obtained from the American Type Culture Collection (Rockville, MD, USA). All the strains were cultured in tryptic soya broth (TSB, OXOID) under aerobic conditions at 37 °C for 24 h on an orbital shaker at 200 rpm. Each tested compound was dissolved in ethanol (Sigma, Milan, Italy) and diluted in TSB to give a stock solution.

Guzzo F., Durán A.G., Sanna C., Marasco R., Molfetta N., Buommino E., Fiorentino A, & D’Abrosca B. (2022). Gallomyrtucommulones G and H, New Phloroglucinol Glycosides, from Bioactive Fractions of Myrtus communis against Staphylococcus Species. Molecules, 27(20), 7109.

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