The sterile filter paper discs (Macherey-Nagel, MN®, Germany), clindamycin (2 μg/disc) by Oxoid (Basingstoke, UK) were used for antibacterial assay. The Brain heart infusion broth, Müller-Hinton broth and agar powder were bought from HiMedia Laboratories Pvt. Ltd. (Mumbai, India). Gas generation kit (AnaeroPack-Anaero) from MITSUBISHI Gas chemical (Tokyo, Japan). 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) and phorbol-12-myristate-13-acetate (PMA) was purchased from Sigma-Aldrich (Sigma Aldrich, Missouri, MO, USA). RPMI-1640 medium, supplemented with 10% fetal bovine serum (FBS) and 1% antibiotic mixture of penicillin G (10,000 units/mL)-Streptomycin (10,000 μg/mL) (Thermo Fisher Scientific, Inc., New York, NY, USA). In addition, analytical grade methanol, 95% ethanol, ethyl acetate, dichloromethane, dimethyl sulfoxide and hexane from RCI Labscan Ltd. (Bangkok, Thailand) were used. A Sandwich ELISA kit was used for pro-inflammatory cytokines detection following the manufacturer’s protocol (BioLegend, Inc., California, CA, USA).
M ller hinton broth
Manufactured by HiMedia
Sourced in India
Müller-Hinton broth is a general-purpose microbiological culture medium used for the cultivation and susceptibility testing of bacteria. It provides the necessary nutrients for the growth of a variety of bacterial species.
Automatically generated - may contain errors
Lab products found in correlation
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide, by Merck Group (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Sandwich elisa kit, by BioLegend (1 mentions)
Phorbol 12 myristate 13 acetate, by Merck Group (1 mentions)
Brain heart infusion broth, by HiMedia (1 mentions)
Methanol, by RCI Labscan (1 mentions)
Clindamycin, by Thermo Fisher Scientific (1 mentions)
Anaeropack anaero, by Mitsubishi (1 mentions)
Agar powder, by HiMedia (1 mentions)
Folin ciocalteu s phenol reagent, by Merck Group (1 mentions)
2 2 diphenyl 1 picrylhydrazyl (dpph), by Merck Group (1 mentions)
Carrageenan, by Merck Group (1 mentions)
Pca agar, by HiMedia (1 mentions)
Peptone, by HiMedia (1 mentions)
4 protocols using m ller hinton broth
1
Antibacterial and Anti-inflammatory Assay Protocol
2
Carrageenan-Based Antioxidant Film Development
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The carrageenan was purchased from Sigma-Aldrich (St. Louis, MO, USA), where the type was κ-carrageenan. The plasticizer used was glycerol, which was purchased from Mistura da Terra (Bagé, Brazil). The following reagents were bought from Sigma-Aldrich (St. Louis, MO, USA) and were of an analytical standard: Folin Ciocalteu’s phenol reagent, 2,2-diphenyl-1-picrylhydrazyl (DPPH), methanol, anhydrous sodium carbonate, gallic acid, and oleuropein. For microbiological analyses: nutrient broth, Müller–Hinton broth, PCA agar, and peptone (Himedia, Bengaluru, India), as well as sterilized distilled water, were used.
3
Antibacterial Activity Screening of Gram-Positive and Gram-Negative Strains
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Antibacterial activity was tested using two Gram negative strains: (i) Aeromonas hydrophila (ATCC 49140) and Shigella flexneri (ATCC 9199); and a panel of Gram positive strains: (ii) Listeria monocytogenes ATCC 19111, Bacillus subtilis ATCC 6633, Enterococcus faecalis ATCC 29212, and Staphylococcus aureus ATCC 25923. The initial screening of antibacterial activity was performed using five more Gram negative strains: (i) Enterobacter cloacae (ATCC 49141), Escherichia coli (ATCC 25922), Proteus mirabilis (ATCC 25933), Pseudomonas aeruginosa (ATCC 15422) and Salmonella enteritidis (ATCC 13076); and two more Gram positive strains: (ii) Micrococcus luteus (ATCC 7468) and Streptococcus equisimilis (ATCC 12394). The bacterial strains were cultured overnight at 37°C in MHB (Müller-Hinton broth, HiMedia, India) with the exception of L. monocytogenes which was cultured in BHI broth (Brain-Heart Infusion, Biomedics, Spain). Suspensions were adjusted to McFarland standard turbidity (0.5) which corresponds to 107–108 CFU/mL.
4
Evaluating OqxA/B Efflux Pumps in E. coli Ciprofloxacin Resistance
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To evaluate the role of OqxA/B efflux pumps in E. coli ciprofloxacin resistance, 5 isolates harboring oqxA/B genes were selected. The reduction in MICs of ciprofloxacin was evaluated using the microdilution method in 96-house plates in accordance with CLSI instructions [62 ]. Briefly, serially twofold dilutions of the ciprofloxacin, with concentrations ranging from 0.12 to 256 µg/mL, were prepared in a sterile Müller Hinton Broth (Himedia, India) culture medium. Then, 100 μL from each dilution was transferred into wells of a microtiter plate and inoculated with 5 µL of standardized bacterial suspension (1.5 × 107 CFU/mL). Finally, a constant volume (4 μL) of the efflux pump inhibitor (PAβN) at a concentration of 100 μg/mL was added to each well. The plates were incubated at 37 °C for 24 h. The change of ciprofloxacin MIC in the presence of the inhibitor was recorded compared to those with no PAβN [66 (link)].
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