Staphylococcus aureus

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Staphylococcus aureus is a bacterial strain available in the American Type Culture Collection (ATCC) product portfolio. It is a Gram-positive, spherical-shaped bacterium commonly found in the human nasal passages and on the skin. This strain is widely used in research and laboratory settings for various applications.

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Microorganism Strains for Antibacterial Studies

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Test microorganism strains that were used in this study include Bacillus cereus ATCC 14579, Bacilus subtilis ATCC 8188, Enterococcus faecalis ATCC 700802, Enterococcus faecalis ATCC 29212, Enterococcus faecalis JH-22, Staphylococcus aureus ATCC 700699, Staphylococcus aureus ATCC 43300, Staphylococcus aureus ATCC 6538P, Staphylococcus aureus ATCC 29213, Aeromonas hydrophila ATCC 49140, Escherichia coli ATCC 25922, Klebsiella pneumoniae ATCC 10031, Proteus mirabilis ATCC 49140, Proteus vulgaris (Institute of Medical Research, Malaysia), Pseudomonas aeruginosa ATCC 10145, Pseudomonas aeruginosa ATCC BAA-47, Salmonella Typhimurium ATCC 14028, and Shigella flexneri ATCC 12022. Strains were cultured on Muller Hinton borth (MHB) (Oxoid, UK) at 37°C and maintained at −80°C in MHB with 25% (v/v) glycerol.

Ong K.S., Aw Y.K., Lee L.H., Yule C.M., Cheow Y.L, & Lee S.M. (2016). Burkholderia paludis sp. nov., an Antibiotic-Siderophore Producing Novel Burkholderia cepacia Complex Species, Isolated from Malaysian Tropical Peat Swamp Soil. Frontiers in Microbiology, 7, 2046.

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Antimicrobial Susceptibility Testing Protocol

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Reference strains of microorganisms that came from American Type Culture Collection (ATCC) (Manassas, VA, USA) were used for this study. The representative Gram-positive bacteria were as follows: Staphylococcus aureus ATCC 25923, Staphylococcus aureus ATCC 29213, Staphylococcus aureus ATCC 43300, Staphylococcus epidermidis ATCC 12228, Enterococcus faecalis ATCC 29212, Micrococcus luteus ATCC 10240, Bacillus subtilis ATCC 6633 and Bacillus cereus ATCC 10876. While, those of Gram-negative bacteria were as follows: Bordetella bronchiseptica ATCC 4617, Escherichia coli ATCC 25922, Klebsiella pneumoniae ATCC 13883, Proteus mirabilis ATCC 12453, Salmonella Typhimurium ATCC 14028 and Pseudomonas aeruginosa ATCC 9027. Moreover, the following fungi belonging to yeasts were used: Candida albicans ATCC 10231, Candida albicans ATCC 2091, Candida parapsilosis ATCC 22019, Candida glabrata ATCC 90030 and Candida krusei ATCC 14243.

Walczak-Nowicka L., Biernasiuk A., Ziemichód W., Karczmarzyk Z., Kwaśnik M., Kozyra P., Wysocki W., Stenzel-Bembenek A., Kowalczuk D., Herbet M, & Pitucha M. (2023). N-Substituted 2-(Benzenosulfonyl)-1-Carbotioamide Derivatives Exert Antimicrobial and Cytotoxic Effects via Aldehyde Dehydrogenase Pathway: Synthesis, In Silico and In Vitro Studies. Pharmaceuticals, 16(12), 1706.

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Antimicrobial Screening of Compounds 1a-1c

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Compounds 1a–c were screened for antibacterial and antifungal activities by microdilution broth method as described elsewhere [24 (link)]. For all compounds, minimal inhibition concentration (MIC), minimal bactericidal concentration (MBC), and minimal fungicidal concentration (MFC) were determined. The tested derivatives were evaluated for the panel of the reference microorganisms from American Type Culture Collection (ATCC), including Gram-positive bacteria (Staphylococcus aureus ATCC 25923, Staphylococcus aureus ATCC 43300, Staphylococcus aureus ATCC 6538, Staphylococcus epidermidis ATCC 12228, Micrococcus luteus ATCC 10240, Bacillus cereus ATCC 10876, Bacillus subtilis ATCC 6633, Streptococcus pyogenes ATCC 19615, Streptococcus mutans ATCC 25175, Streptococcus pneumoniae ATCC 49619),Gram negative bacteria (Salmonella Typhimurium ATCC14028, Escherichia coli ATCC 25922, Klebsiella pneumoniae ATCC 13883, Pseudomonas aeruginosa ATCC 9027, Proteus mirabilis ATCC 12453), and fungi (Candida parapsilosis ATCC 22019, Candida albicans ATCC 10231, Candida parapsilosis ATCC 22019) (LGC Standards, Teddington, Middlesex, UK). Each experiment was performed in triplicate. Representative data are presented.

Gobis K., Szczesio M., Olczak A., Mazerant-Politowicz I., Ziembicka D., Pacholczyk-Sienicka B., Augustynowicz-Kopeć E., Głogowska A., Korona-Głowniak I, & Fruziński A. (2022). N′-Substituted 4-Phenylpicolinohydrazonamides with Thiosemicarbazone Moiety as New Potential Antitubercular Agents: Synthesis, Structure and Evaluation of Antimicrobial Activity. Materials, 15(16), 5513.

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Antimicrobial Efficacy Evaluation

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Ten bacterial strains from American Type Culture Collection (ATCC, Rockville, MD, USA) were used for tests: Bacillus cereus ATCC 10876, Bacillus subtilis ATCC 6633, Staphylococcus aureus ATCC 6538p, Staphylococcus aureus ATCC 29213, Staphylococcus aureus ATCC 43300 (MRSA), Enterococcus faecalis ATCC 29212, Escherichia coli ATCC 25922, Klebsiella pneumoniae ATCC 19883, Acinetobacter baumanni ATCC 19606, Pseudomonas aeruginosa ATCC 27853. Seven Candida species from ATCC were used for antifungal tests: Candida albicans ATCC 10231, Candida albicans ATCC 24433, Candida albicans ATCC 90028, Candida guilliermondii ATCC 6260, Candida glabrata ATCC 15126, Candida krusei ATCC 14243, Candida kefyr ATCC 204093.

Rosato A., Carocci A., Catalano A., Clodoveo M.L., Franchini C., Corbo F., Carbonara G.G., Carrieri A, & Fracchiolla G. (2018). Elucidation of the synergistic action of Mentha Piperita essential oil with common antimicrobials. PLoS ONE, 13(8), e0200902.

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Antimicrobial Susceptibility Testing Strains

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The reference strains of microorganisms from American Type Culture Collection (ATCC), Manassas, VA, USA, were included. The representative Gram-positive bacteria were: Staphylococcus aureus ATCC 6538, Staphylococcus aureus ATCC 25923, Staphylococcus epidermidis ATCC 12228, Micrococcus luteus ATCC 10240, Bacillus subtilis ATCC 6633 and Bacillus cereus ATCC 10876, while those of Gram-negative bacteria: Escherichia coli ATCC 25922, Proteus mirabilis ATCC 12453, Klebsiella pneumoniae ATCC 13883, Salmonella Typhimurium ATCC 14028, Pseudomonas aeruginosa ATCC 9027 and Bordetella bronchiseptica ATCC 4617. Moreover, the fungi belonging to yeasts (Candida albicans ATCC 2091, Candida albicans ATCC 10231, Candida parapsilosis ATCC 22019, Candida glabrata ATCC 90030 and Candida krusei ATCC 14243) were used.

Biernasiuk A., Banasiewicz A., Masłyk M., Martyna A., Janeczko M., Baranowska-Łączkowska A., Malm A, & Łączkowski K.Z. (2021). Synthesis and Physicochemical Characterization of Novel Dicyclopropyl-Thiazole Compounds as Nontoxic and Promising Antifungals. Materials, 14(13), 3500.

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Antimicrobial Activity Screening Protocol

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The microdilution method was used to determine the antimicrobial activity according to the European Committee on Antimicrobial Susceptibility Testing [20 ]. MH broth and MH broth with 2% glucose were used to grow the bacteria and yeasts, respectively. The following microbial strains were tested: Bacillus subtilis ATCC 6633, Candida albicans ATCC 2091, Candida glabrata ATCC 90030, Candida parapsilosis ATCC 22019, Enterococcus faecalis ATCC 29212, Escherichia coli ATCC 25922, Klebsiella pneumoniae ATCC 13883, Micrococcus luteus ATCC 10240, Proteus mirabilis ATCC 12453, Pseudomonas aeruginosa ATCC 90271, Salmonella Typhimurium ATCC 14028, Staphylococcus aureus ATCC 25923, Staphylococcus aureus ATCC BAA-1707 (methicillin-resistant strain), and Staphylococcus epidermidis ATCC 12228. The minimum inhibitory concentration (MIC) was calculated and reported for each sample and strain. All experiments were performed in triplicate.

Luca S.V., Zengin G., Sinan K.I., Korona-Glowniak I., Minceva M., Skalicka-Woźniak K, & Trifan A. (2023). Value-Added Compounds with Antimicrobial, Antioxidant, and Enzyme-Inhibitory Effects from Post-Distillation and Post-Supercritical CO2 Extraction By-Products of Rosemary. Antioxidants, 12(2), 244.

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Antibacterial Assay and MIC Determination

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Bacterial strains including Gram-positive bacteria (Enterococcus faecalis ATCC 29212, Micrococcus luteus DMST 15503, methicillin resistant Staphylococcus aureus NPRC 001R, Staphylococcus aureus ATCC 25923, Streptococcus pyogenes ATCC, 19615), Gram-negative bacteria (Shigella flexneri DMST 4423, Pseudomonas aeruginosa ATCC 10145, Salmonella typhi DMST 22842, Salmonella typhimurium DMST 562, Escherichia coli TISTR 780) and fungi (Candida albicans ATCC 10231) used in these experiments were obtained from the Microbiological Resources Center, Thailand. A 2-fold serial dilution method (Nutrient broth) was used for the antibacterial assays and the determination of minimum inhibitory concentrations (MICs) (Wikler et al., 2006 ; Tantapakul et al., 2016 (link)). The antimicrobial assays were tested in triplicate, and the standard compounds were vancomycin and gentamycin.

Phukhatmuen P., Meesakul P., Suthiphasilp V., Charoensup R., Maneerat T., Cheenpracha S., Limtharakul T., Pyne S.G, & Laphookhieo S. (2021). Antidiabetic and antimicrobial flavonoids from the twigs and roots of Erythrina subumbrans (Hassk.) Merr.. Heliyon, 7(4), e06904.

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Microbial and Cell Line Sources

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Escherichia coli (ATCC^® 8739), Staphylococcus aureus (ATCC^® 6538), Staphylococcus aureus (ATCC^® 29737), Bacillus subtilis (ATCC^® 6633), Salmonella enterica sv typhimurium (ATCC^® 14028), Salmonella species sv Abony (NCTC 6017), Kocuria rhizophila (ATCC^® 9341), Candida albicans (ATCC^® 10231), Aspergillus brasiliensis (ATCC^® 16404), Human Breast Adenocarcinoma (MCF-7 ATCC^® HTB-22TM) and Human Lung Adenocarcinoma (SK-LU-1; ATCC^® HTB-57TM) cell lines were obtained from the American Type Culture Collection (ATCC).

Rahaman M.S., Siraj M.A., Sultana S., Seidel V, & Islam M.A. (2020). Molecular Phylogenetics and Biological Potential of Fungal Endophytes From Plants of the Sundarbans Mangrove. Frontiers in Microbiology, 11, 570855.

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Antimicrobial Evaluation of Novel Compounds

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The microbial strains used were provided by King Saud Medical City Central Laboratories. Bacterial strains include gram-positive bacterial strains Staphylococcus aureus (ATCC 25923) used for agar-well diffusion, Staphylococcus aureus (ATCC 29213) for determining the minimum inhibitory concentration, Staphylococcus epidermidis (ATCC 12228), and Enterococcus faecalis (ATCC 29212). Gram-negative strains included Escherichia coli (ATCC 25922), Pseudomonas aeruginosa (ATCC 27853), and Klebsiella pneumoniae (ATCC 700603). Fungal strains were additionally tested and included Candida albicans (ATCC 10231) and Candida parapsilosis (ATCC 22019). All strains were cultured on Mueller Hinton agar and broth and then adjusted to 0.5 McFarland turbidity in 10 mL Mueller Hinton broth for antimicrobial assays.

Elmongy E.I., Alanazi W.S., Aldawsari A.I., Alfaouri A.A, & Binsuwaidan R. (2024). Antimicrobial Evaluation of Sulfonamides after Coupling with Thienopyrimidine Coplanar Structure. Pharmaceuticals, 17(2), 188.

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Antimicrobial Evaluation of Pathogenic Strains

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The following strains from the University of Messina's in-house culture collection (Messina, Italy) were used for the antimicrobial testing: Staphylococcus aureus ATCC 6538P, Staphylococcus aureus MRSA ATCC 43300, Staphylococcus epidermidis ATCC 49134, Staphylococcus epidermidis ATCC 35984, Staphylococcus epidermidis ATCC 12228, Streptococcus pneumoniae ATCC 6003, Streptococcus pyogenes ATCC 19615, Streptococcus mutans ATCC 35668, Listeria monocytogenes ATCC 7644, Listeria monocytogenes ATCC 1392, Enterococcus hirae ATCC 10541, Moraxella catarrhalis ATCC 8176, Bacillus subtilis ATCC 8176, Enterococcus durans (wild-type strain), Escherichia coli ATCC 25922, Klebsiella pneumoniae (wild-type strain), Pseudomonas aeruginosa ATCC 27853, Pseudomonas aeruginosa ATCC 9027, Pseudomonas aeruginosa ATCC 15442, Proteus mirabilis (wild-type strain), Serratia marcescens (wild-type strain), Salmonella typhi ATCC 13311, Candida albicans ATCC 10231, Candida parapsilosis ATCC 29947, and the fungus Aspergillus niger ATCC 16404.
Bacteria were grown in Mueller-Hinton Broth (MHB, Oxoid, CM0405) at 37°C (24 h), whereas yeasts were cultured in Sabouraud Liquid Medium (SLM, Oxoid, CM0147) at 30°C (48 h). For solid media, 1.5% (w/v) agar (Difco) was added. Aspergillus niger was grown in Sabouraud Dextrose Agar at 30°C for 7 days as previously reported [10 (link)].

Filocamo A., Bisignano C., Mandalari G, & Navarra M. (2015). In Vitro Antimicrobial Activity and Effect on Biofilm Production of a White Grape Juice (Vitis vinifera) Extract. Evidence-based Complementary and Alternative Medicine : eCAM, 2015, 856243.

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