Chemical UK) and fluconazole (> 98%, Acros Organics UK) were conducted to assess the adsorption capacity in the concentration range of 15-320 mg L -1 (fluconazole) and 5-500 mg L -1 (caffeine). For the adsorption experiments, 25 mg of biochar (wt. % d.b.) were mixed with 25 mL of pharmaceutical solution, shaken for 24 hours on an orbital shaker at 180 rpm before filtering through 0.45 µm hydrophilic PTFE syringe filters. All experiments were conducted at room temperature of 20 °C. The supernatants were analysed by UV-Vis spectrophotometry at 260 nm for both contaminants, with linear calibration ranges of 10-100 mg L -1 (fluconazole) and 2-125 mg L -1 (Caffeine). Equilibrium concentrations were calculated according to Yan et al. (2015) (link). The obtained isotherm data was fitted using the non-linear forms of the Langmuir and Freundlich isotherm models to calculate the adsorption capacity (Kumar, 2006) (link). Characterisation of the softwood and wheat straw pellets highlights the main differences between the two feedstocks, with ash contents of 1.9 % (SWP) and 7.9 % (WSP) and a lower FC content for softwood than wheat straw (8.6 % and 15.2 %).
Fluconazole
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom, United States, Germany
Fluconazole is a pharmaceutical compound used in the treatment of fungal infections. It is an antifungal agent that inhibits the synthesis of ergosterol, a critical component of fungal cell membranes. Fluconazole is commonly prescribed for the management of conditions such as candidiasis, cryptococcosis, and other fungal infections.
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Lab products found in correlation
Chloramphenicol, by Thermo Fisher Scientific (7 mentions)
Sabouraud dextrose agar, by Thermo Fisher Scientific (5 mentions)
Mueller hinton agar (mha), by Thermo Fisher Scientific (5 mentions)
Gentamicin, by Thermo Fisher Scientific (4 mentions)
Voriconazole, by Tokyo Chemical Industry (4 mentions)
Cefoxitin, by Thermo Fisher Scientific (3 mentions)
Caspofungin, by Merck Group (3 mentions)
Rpmi 1640, by Thermo Fisher Scientific (3 mentions)
Itraconazole, by Tokyo Chemical Industry (3 mentions)
Gentamicin sulfate, by Chem-Impex (3 mentions)
Amphotericin b, by Thermo Fisher Scientific (2 mentions)
Dimethylsulfoxid, by Centralchem (2 mentions)
Rpmi powder, by Merck Group (2 mentions)
Flucytosine, by Tokyo Chemical Industry (2 mentions)
Nile red, by Tokyo Chemical Industry (2 mentions)
Triton x 100, by Thermo Fisher Scientific (2 mentions)
Voriconazole, by Thermo Fisher Scientific (2 mentions)
Mueller hinton agar, by Thermo Fisher Scientific (2 mentions)
Dimethyl sulfoxide (dmso), by Thermo Fisher Scientific (2 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
37 protocols using fluconazole
1
Biochar Adsorption of Fluconazole and Caffeine
2
Antimicrobial Evaluation of Grape Pomace
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The agar disc diffusion method was used for the determination of the antimicrobial activity of the grape pomace extracts. Briefly, a suspension of the tested microorganism (0.1 mL of 105 cells/mL) was spread onto Mueller Hinton Agar (MHA, Oxoid, Basingstoke, UK) and Sabouraud dextrose agar (Oxoid, Basingstoke, UK) at 25 °C. Filter paper discs (6 mm in diameter) were impregnated with 15 µL of the grape pomace extract and placed on the inoculated plates. Chloramphenicol (10 μg/disk, Oxoid, Basingstoke, UK) for G−, streptomycin (10 μg/disk, Oxoid, Basingstoke, UK) for G + and Fluconazole (25 µg/disc, Oxoid, Basingstoke, UK) for yeasts were used as a positive control to determine the sensitivity of the microorganisms under study. The plates were kept at 4 °C for 2 h and subsequently incubated aerobically at 37 °C for 24 h and 25 °C for 24 h for bacteria and yeast, respectively. The diameters of the inhibition zones were measured in millimetres. All the tests were performed in triplicate.
3
Antimicrobial Activity of Usnea barbata Extracts
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Usnea barbata dry extracts and usnic acid were dissolved in 0.1% DMSO and applied on Whatman® filter paper discs (6 mm, Merk KGaA, Darmstadt, Germany). The solvent (0.1% DMSO) was the negative control and UA in 0.1% DMSO (129 mg/mL) was the positive control for UBDE [46 (link)]. The weighted mass values for UA and each UBDE were similar to those used in our previous study to evaluate their cytotoxic activity by brine shrimp lethality assay [46 (link)]. Therefore, the concentrations of the sample UBDE solutions were as follows: 172 mg/mL UBEA, 162 mg/mL UBA, 161 mg/mL UBE and UBM, and 160 mg/mL UBW [46 (link)]. Each filter paper disc was impregnated with 10 µL solution. For antimicrobial activity evaluation, blank antibiotic discs (6 mm)—Levofloxacin 5 µg, Tetracycline 30 µg, and antifungal ones—Voriconazole 1 µg and Fluconazole 15 µg (Oxoid, Thermo Fisher Scientific GmbH, Dreieich, Germany) were used. The blank discs were maintained in a freezer at −14 °C and incubated at room temperature for 2 h before analysis.
Each inoculum was applied with a sterile cotton swab over the entire surface of the plate with the suitable culture media. After 15 min of drying, the filter paper discs were applied to the inoculated plates. The plates were incubated for 24 h at 37 °C.
Each inoculum was applied with a sterile cotton swab over the entire surface of the plate with the suitable culture media. After 15 min of drying, the filter paper discs were applied to the inoculated plates. The plates were incubated for 24 h at 37 °C.
4
Antimicrobial Efficacy of Essential Oils
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The antibacterial activity was examined by the agar disc diffusion method. S. maltophilia and B. subtilis were incubated in Mueller Hinton broth (MHB, Oxoid, Basingstoke, UK) at 37 °C for 24 h. P. expansum, P. crustosum, and P. citrinum were incubated in Sabouraud broth (SB, Oxoid, Basingstoke, UK) at 25 °C for 48 h. The isolates were identified by useof the MALDI-TOF MS Biotyper (Bruker Daltonic, Bremen, Germany) with a score higher than 2.2. Microbial suspensions were prepared in saline and adjusted to 0.5 McFarland turbidity standards with a densilameter (Erba Lachema s.r.o., Brno, Czech Republic). The isolates were used for inoculation onto the Mueller–Hinton agar (MHA) and Sabouraud agars, subsequently discs impregnated by the tested EOs (10 µL/disc) were used for the detection of the antimicrobial activity. MHA were incubated at 4 °C for 1–2 h, and then at 37 °C and 25 °C for 18–24 h and 48 h for bacteria and microscopic fungi, respectively. The antimicrobial activity was evaluated by measuring the zone of growth inhibition around the discs following incubation. Chloramphenicol (30 µg, Oxoid, Basingstoke, UK) and fluconazole (25 µg, Oxoid, Basingstoke, UK) served as positive antimicrobial controls.
5
Antimicrobial Activity of H. italicum Essential Oil
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The EO of H. italicum (plant material originated in Bosnia, France and Corsica) was tested against nine microorganisms with an agar disc diffusion method according to in our previous study [66 (link)]. In this study, 0.1 mL of microbial suspension was spread on the Mueller Hinton Agar (MHA, Oxoid, UK) for bacteria and Sabouraud Dextrose agar (SDA, Oxoid, UK) for yeasts. Six mm diameter filter paper discs were used for testing. The filter paper was impregnated with 15 μL of EO and placed on MHA, SDA, respectively, with a microbial inoculum. The MHA was maintained at 4 °C for 2 h and then at 37 °C for 24 h and SDA was maintained at 4 °C for 2 h and then at 25 °C for 24 h. After a 24 h incubation period, the diameter of the inhibition zones was measured (in mm). Chloramphenicol (30 µg, Oxoid, UK) and fluconazole (25 µg, Oxoid, UK) served as positive antimicrobial controls. Antimicrobial activity was measured in triplicate.
6
Antimicrobial Activity of CBPR Films
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The antimicrobial activity of CBPR films was measured by Kirby–Bauer assay in MHA (Gram-positive S. aureus and S. epidermidis, Gram-negative P. aeruginosa) or in SDA Agar (Candida) Petri dishes.
Microbial suspensions were prepared from an 18 h culture in a liquid medium. The microorganisms were recovered in sterile phosphate buffer (PBS) and brought to a concentration of 108 CFU/mL.
Dishes were seeded with the different microbial strains by sliding (rotating the plate 60° three times). The 6 mm diameter UV-sterilized film disks were hydrated with 20 μL of sterile water to facilitate both the placement on the surface of Petri dishes seeded with the microorganisms and to promote the release of the active ingredients from the film. A film of only chitosan was used as a negative control. Filter paper disks containing gentamicin (30 μg, Oxoid) for bacterial strains and fluconazole (25 μg, Oxoid) for C. albicans were used as positive controls. After 24 h of incubation at 37 °C, the microbial growth inhibition halos were measured. The extent of inhibition (80% growth reduction) was expressed as the diameter of the halo zone in mm.
Microbial suspensions were prepared from an 18 h culture in a liquid medium. The microorganisms were recovered in sterile phosphate buffer (PBS) and brought to a concentration of 108 CFU/mL.
Dishes were seeded with the different microbial strains by sliding (rotating the plate 60° three times). The 6 mm diameter UV-sterilized film disks were hydrated with 20 μL of sterile water to facilitate both the placement on the surface of Petri dishes seeded with the microorganisms and to promote the release of the active ingredients from the film. A film of only chitosan was used as a negative control. Filter paper disks containing gentamicin (30 μg, Oxoid) for bacterial strains and fluconazole (25 μg, Oxoid) for C. albicans were used as positive controls. After 24 h of incubation at 37 °C, the microbial growth inhibition halos were measured. The extent of inhibition (80% growth reduction) was expressed as the diameter of the halo zone in mm.
7
Fluconazole Antifungal Susceptibility Testing
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Antifungal susceptibility testing of fluconazole (25 μg) (Oxoid Ltd., Basingstoke, UK) was carried out by disc diffusion method as described by Clinical and Laboratory Standards Institute (CLSI, document M44-A2, 2009). A zone diameter of ≥19 mm was considered sensitive, 15 to 18 mm was considered dose-dependently susceptible, and a diameter ≤14 mm was considered resistant [21 (link)].
8
Antimicrobial Efficacy Evaluation via Disc Diffusion
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Antimicrobial activity was determined by using the disc diffusion method
[21 (link)]. All samples (dry residue) were dissolved in 10% sterile dimethyl sulfoxide. The discs (6 mm diameter) were impregnated with 10 mg/mL extract/fractions (100 μL/disc) placed aseptically on the inoculated agar. Discs injected with 100 μL of respective solvents served as a negative controls, rifampcin (100 μL/disc) (Oxoid) and fluconazole (100 μL/disc) (Oxoid) were used as positive reference for bacteria and fungi, respectively. The Petri dishes were incubated at 37 ± 0.1°C for 20–24 h and 28 ± 0.3°C for 40–48 h for bacteria and fungi, respectively. At the end of period, the inhibition zones formed on the media were measured. The positive antimicrobial activity was read based on growth inhibition zone.
[21 (link)]. All samples (dry residue) were dissolved in 10% sterile dimethyl sulfoxide. The discs (6 mm diameter) were impregnated with 10 mg/mL extract/fractions (100 μL/disc) placed aseptically on the inoculated agar. Discs injected with 100 μL of respective solvents served as a negative controls, rifampcin (100 μL/disc) (Oxoid) and fluconazole (100 μL/disc) (Oxoid) were used as positive reference for bacteria and fungi, respectively. The Petri dishes were incubated at 37 ± 0.1°C for 20–24 h and 28 ± 0.3°C for 40–48 h for bacteria and fungi, respectively. At the end of period, the inhibition zones formed on the media were measured. The positive antimicrobial activity was read based on growth inhibition zone.
9
Candida Antifungal Susceptibility Testing
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Antifungal susceptibility testing was performed for all Candida isolates using a modified disc diffusion method as per Clinical Laboratory Standard Institute guideline by adding 2% glucose and 0.5 μg/mL Methylene Blue dye into Mueller-Hinton agar. A suspension was prepared using normal saline by adding five different colonies and incubating overnight in Sabouraud dextrose agar. Then the suspension was compared to 0.5 McFarland standards. Cotton swab moistened with the fungal suspension was streaked on modified Mueller-Hinton media. Antifungal discs including amphotericin B 100 μg, clotrimazole10μg, fluconazole25 μg, itraconazole10 μg, and ketoconazole10 μg (Oxoid, UK), were placed on Mueller-Hinton agar using disk dispenser, and the plates were incubated at 370C for 24 h. Finally, the zones of inhibition (Zone diameters) were measured and interpreted according to CLSI guideline (M44-A2).
10
Antifungal Activity of Imidazole Derivatives
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Preliminary tests regarding the inhibitory activity of compounds were performed using the agar diffusion method. Several colonies from 24 h old cultures grown on Sabouraud Dextrose Agar (SDA) were suspended in sterile saline solution and adjusted to 1–5 × 105 CFU/mL. Inoculations were made via flooding the SDA surface with 1 mL of fungal suspension. The excess of suspension was removed and plates were kept inverted to allow the agar to dry. Afterward, wells were punched on agar plates (5 mm in diameter), and 50 µL of each compound was pipetted into the wells. The plates were incubated at 37 °C for 48 h, and antifungal activity was assessed as mm of inhibition zones. In order to compare the susceptibility of Candida spp. to imidazole derivatives SAM3 (200 µg), AM5 (300 µg) and SAM5 (300 µg), we used fluconazole (25 µg, Oxoid—Basingstoke, UK), an antifungal widely used in the treatment of infections caused by Candida spp., as a positive control. The testing and evaluation of the strains’ susceptibility were performed according to the CLSI standards [23 ].
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