Each P. aeruginosa strain was cultured in LB medium to logarithmic phase at 37°C with rotary shaking at 150 rpm (Excella E-24 incubator shaker; Eppendorf, Hamburg, Germany). The culture was centrifuged at 10,000 × g for 2 min and the bacterial pellet was washed three times with physiological saline. The pellet was resuspended in an adequate volume using physiological saline, and the optical density (OD) at 600 nm (OD 600 ) was adjusted to give the approximate desired inocula (OD 600 of 1 ≅ 5 × 10 8 colony-forming units [CFU]/mL). The inocula were verified by serial 10-fold dilutions of the suspensions and plating on cetrimide agar (Nissui Pharmaceutical, Tokyo, Japan).
Cetrimide agar
Manufactured by Nissui Pharmaceutical
Sourced in Japan
Cetrimide agar is a microbiological culture medium used for the selective isolation and identification of Pseudomonas species from clinical and environmental samples. It contains cetrimide, a quaternary ammonium compound that inhibits the growth of most other bacteria, allowing Pseudomonas species to grow selectively.
Automatically generated - may contain errors
3 protocols using cetrimide agar
1
Inoculum Preparation for P. aeruginosa
2
Bacterial Reactivity Comparison of Oral Gels
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The control gel and MA-T gel were used to compare reactivity to bacteria. Most gram-positive and gram-negative bacteria were cultured on cetrimide agar (Nissui, Tokyo, Japan). MRSA and P. aeruginosa-selective media were used for culture of the respective organisms. Oral film of 0.5 g and 1.0 mL of sterilized distilled pure water were mixed by vortex for 1 min after being left for 10 min at 28 °C. Half of the liquid part of the mixed oral epithelium was mixed with gel and spread in the culture medium prepared in 9-cm diameter plates. Three experiments were performed. After culturing the cells for 24 h, the number of bacteria was evaluated.
3
Antimicrobial Activity of Fly Larvae
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Musca domestica larvae were provided by E's Inc. (Tokyo, Japan). Staphylococcus aureus (NBRC100910), Staphylococcus epidermidis (NBRC100911), and Pseudomonas aeruginosa (NBRC12689) were purchased from National Institute of Technology and Evaluation (Kisarazu, Chiba, Japan). Escherichia coli XL1-blue was purchased from Stratagene (Agilent Technologies, Santa Clara, California, USA). Mannitol salt agar and cetrimide agar were purchased from Nissui Pharmaceutical (Tokyo, Japan). Mueller-Hinton II broth was purchased from Becton Dickinson (Franklin Lakes, New Jersey, USA). LB broth was purchased from Nacalai tesque (Kyoto, Japan). Agar for bacterial culture was purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan).
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