S. aureus (ATCC 25923), B. subtilis (ATCC 11774), B. cereus (ATCC 14579), E. coli (ATCC 25922), P. aeruginosa (ATCC 15442), S. typhimurium (ATCC 14028), C. lusitaniae (ATCC 34449), and A. niger (ATCC 16888) strains were provided by the Affiliated Hospital of Qingdao University. CPC, fetal calf serum (FCS), Eagle’s minimum essential medium (EMEM), and agar powder were purchased from Sangon Biotech Co., Ltd. (Shanghai, China). CHA and bovine serum protein (BSA) were purchased from Solarbio Science & Technology Co., Ltd. (Beijing, China). LB broth medium, Sabouraud dextrose broth medium, and D/E (Dey/Engley) neutralizing broth were purchased from Hope Bio-Technology Co., Ltd. (Qingdao, China). Glutaraldehyde solution (2.5%, v/v) was purchased from Macklin Biochemical Co., Ltd. (Shanghai, China). Filmtracer™ LIVE/DEAD™ Biofilm Viability Kit was purchased from Thermo Fisher Scientific (China) Co., Ltd. (Shanghai, China). BCA Protein Colorimetric Assay Kit was purchased from Elabscience Biotechnology Co., Ltd. (Wuhan, China). New Zealand white rabbits were purchased from Lukang Pharmaceutical Co., Ltd (Qingdao, China). All the other chemicals and reagents were of analytical grade.
Agar powder
Manufactured by Sangon
Sourced in China
Agar powder is a natural polysaccharide derived from red algae. It is commonly used as a gelling agent in microbiological culture media, allowing for the solidification of nutrient broths to create agar plates. Agar powder is a versatile ingredient, providing a stable, transparent, and thermoreversible gel when dissolved in water.
Automatically generated - may contain errors
Lab products found in correlation
Sodium chloride (nacl), by Sangon (3 mentions)
Yeast extract, by Thermo Fisher Scientific (2 mentions)
Bacto tryptone, by Thermo Fisher Scientific (2 mentions)
Filmtracer live dead biofilm viability kit, by Thermo Fisher Scientific (1 mentions)
Bca protein colorimetric assay kit, by Elabscience (1 mentions)
Glutaraldehyde solution, by Maclin Biochemical Technology (1 mentions)
Hydrogen peroxide, by Sangon (1 mentions)
Ferrous sulfate, by Sangon (1 mentions)
Sodium bicarbonate, by Sangon (1 mentions)
Acetone, by Sangon (1 mentions)
Congo red, by Sangon (1 mentions)
Tryptone, by Sangon (1 mentions)
Mueller hinton broth, by Thermo Fisher Scientific (1 mentions)
5 protocols using agar powder
1
Antimicrobial Activity Evaluation Protocol
2
Rhodotorula glutinis SRY Strain Cultivation
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The Rhodotorula glutinis SRY strain was isolated and maintained by the Food Microbiology Team, Agro-Processing Institute, Jilin Academy of Agricultural Sciences. Protein, yeast extract powder, agar powder, sodium chloride, glucose, concentrated hydrochloric acid, acetone, zinc sulfate, manganese sulfate, ferrous sulfate, copper sulfate, hydrogen peroxide and sodium bicarbonate were all purchased from Sangon Biotech (Shanghai) Co., Ltd.; vitamin B1 was purchased from Cisen Pharmaceutical Co., Ltd.; soybean oil was purchased from Yihai Kerry Arawana Holdings Co, Ltd. and TWW is taken from the local market.
3
Isolation and Characterization of Multi-Drug Resistant E. coli
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The multi-drug resistant E. coli strain ECDCM1 used in this study was isolated from a dairy cow with mastitis. Strain ECDCM1 was stored at −80 °C. Before each experiment, it was first cultured on Luria-Bertani (LB) agar plates which contained 10 g/L Bacto Tryptone (Oxoid, Basingstoke, UK), 5 g/L yeast extract (Oxoid, Basingstoke, UK), 10 g/L NaCl (Sangon, Shanghai, China), and 20 g/L agar powder (Sangon, Shanghai, China) for 16 h at 37 °C in air supplied with 5% CO2. QA NPs were synthesized according to a previous study (Sun et al., 2017 (link)).
4
Examining ompR Gene's Impact on Bacterial Growth
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To determine the effect of the ompR gene on bacterial growth, the AE17, AE17ΔompR, and AE17C-ompR strains were determined on LB medium. Briefly, bacteria were incubated in LB medium at 37°C and 150 RPM, and the optical density (OD ) of strains was monitored at 1 h intervals by spectrophotometry (Bio-Rad).
Overnight cultures of AE17, AE17ΔompR, and AE17C-ompR strains were diluted 1:25 and statically cultured to the logarithmic phase. The bacteria were washed with phosphate buffer saline (PBS ), plated on the LB semi-solid medium (0.25% agar), and incubated at 37°C for 8 h, then the motile cycles of the strains were observed. The LB semi-solid medium was prepared from 0.5% NaCl, 1% tryptone, 0.8% glucose, and 0.25% agar powder (Sangon, Shanghai, China).
Congo red plates were used to detect curli production. The overnight bacteria were added to Congo red plates and incubated at 37°C for 3 d. Congo red plates were LB agar plate without salt supplemented with 40 mg/L Congo red (Sangon) and 20 mg/L brilliant blue (Sangon).
Overnight cultures of AE17, AE17ΔompR, and AE17C-ompR strains were diluted 1:25 and statically cultured to the logarithmic phase. The bacteria were washed with phosphate buffer saline (
Congo red plates were used to detect curli production. The overnight bacteria were added to Congo red plates and incubated at 37°C for 3 d. Congo red plates were LB agar plate without salt supplemented with 40 mg/L Congo red (Sangon) and 20 mg/L brilliant blue (Sangon).
5
Isolation and Cultivation of Mastitis-Derived E. coli
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The E. coli strain used in this study was isolated from a dairy cow with mastitis. The E. coli was stored at -80°C. Before each experiment, it was first cultured on Luria broth agar plates that contained 10 g/L of Bacto tryptone (Oxoid, Basingstoke, UK), 5 g/L of yeast extract (Oxoid), 10 g/L of NaCl (Sangon, Shanghai, China), and 20 g/L of agar powder (Sangon) for 16 h at 37°C in air supplemented with 5% CO 2 . Colonies were then cultivated overnight in 2 mL of Mueller-Hinton (MH) broth (Oxoid), designated as the first overnight cultures, and subsequently E. coli from the first overnight culture was diluted to an optical density at 600 nm of approximately 0.03 in fresh MH broth for the following experiments. Cultures of the E. coli strain were grown at 37°C with shaking at 200 rpm.
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