Antibiotic sensitivity to cefiderocol (MedChemExpress, Monmouth Junction, NJ, USA) and minimum inhibitory concentrations (MIC) for piperacillin/tazobactam, ciprofloxacin, cefepime, meropenem, and imipenem (Liofilchem, Waltham, MA, USA) were obtained by recording the lowest concentration of antibiotic that inhibited growth of bacteria on Mueller Hinton agar (Neogen, Lansing, MI, USA), as described by the manufacturer’s protocol.
Ciprofloxacin
Manufactured by Liofilchem
Sourced in Italy
Ciprofloxacin is a fluoroquinolone antibiotic used in laboratory settings. It is a broad-spectrum antimicrobial agent effective against a variety of bacterial pathogens.
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Lab products found in correlation
Gentamicin, by Liofilchem (10 mentions)
Ceftazidime, by Liofilchem (9 mentions)
Imipenem, by Liofilchem (7 mentions)
Tetracycline, by Liofilchem (7 mentions)
Amikacin, by Liofilchem (6 mentions)
Cefepime, by Liofilchem (5 mentions)
Cefoxitin, by Liofilchem (5 mentions)
Amoxicillin, by Liofilchem (4 mentions)
Cefotaxime, by Liofilchem (4 mentions)
Meropenem, by Liofilchem (3 mentions)
Aztreonam, by Liofilchem (3 mentions)
Nitrofurantoin, by Liofilchem (3 mentions)
Nalidixic acid, by Liofilchem (3 mentions)
Erythromycin, by Liofilchem (3 mentions)
Chloramphenicol, by Liofilchem (2 mentions)
Fosfomycin, by Liofilchem (2 mentions)
Ampicillin, by Liofilchem (2 mentions)
Oxacillin, by Liofilchem (2 mentions)
Trimethoprim sulfamethoxazole, by Liofilchem (2 mentions)
Cefepime, by bioMérieux (2 mentions)
16 protocols using ciprofloxacin
1
Antibiotic Sensitivity and MIC Determination
2
Antibiotic Susceptibility Testing of E. coli
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A total of 18 antibiotics were used in the susceptibility test, by disk diffusion, with amoxicillin, amoxicillin and clavulanic acid, cefoxitin, cefotaxime, ceftazidime, ceftiofur, cefepime, ceftaroline, aztreonam, meropenem, ciprofloxacin, gentamicin, sulfamethoxazole-trimethoprim, tigecycline, tetracycline, fosfomycin, chloramphenicol, and nitrofurantoin (Liofilchem, Roseto degli Abruzzi, Italy). The interpretation was performed according to CLSI [45 ]. tigecycline resistance was interpreted according to EUCAST [46 ]. E. coli ATCC 25922 was used as quality control. In accordance with EUCAST standards, the minimal inhibitory concentration (MIC) for colistin was determined by the broth microdilution method [46 ]. MDR profiles were determined according to standard criteria [26 (link)]. ESBL detection was performed using the double-disk synergy test (DDST) [45 ].
3
Antibiotics and Artemisinin Derivatives Sensitivity
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Artemisinin, Dihydroartemisinin, Artesunate, and Artemisinic acid were obtained from the China Institute of food and drug control and dissolved following the instruction before use. All antibiotics (ciprofloxacin, imipenem, penicillin G, ampicillin, and oxacillin) were purchased from the North China Pharmaceutical Group Corp (Shijiazhuang, China) and Southwest Synthetic Pharmaceutical Co. Ltd (Chongqing, China). All antibiotics were dissolved and diluted according to Clinical and Laboratory Standards Institute (CLSI) guidelines (23 ). Antibiotics disks (10 μg) of all five antibiotics and strips of ciprofloxacin, imipenem (0.002–32 mg), penicillin G, ampicillin, and oxacillin (0.016–256 mg) were purchased from Liofilchem srl (Zona Industriale Italy). All standard strains of E. coli BNCC 186347, 125787, 125988, and 195617 were purchased from BeNe Culture Collection, Kunshan city, Jiangsu province, China.
4
Antimicrobial Susceptibility Testing Protocol
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The Kirby Bauer disk diffusion method was used with Muller Hinton agar (Oxoid Ltd. Basingstoke, Hampshire, England) to determine antimicrobial susceptibility patterns of the isolates and CLSI M100 2020 was used to interpret the results [30 ].
The following antimicrobial discs were used: ampicillin (10μl), amoxicillin/clavulanic acid (20/10μl), piperacillin/tazobactam (100/10μl), cefazolin (30μl), cefuroxime (30μl), ceftazidime (30μl) obtained from Hardy Diagnostics, Santa Maria, CA, USA. Ceftriaxone (30μl), cefotaxime (30μl), cefepime (30μl), imipenem (10μl), meropenem (10μl), amikacin (30μl), gentamicin (10μl), and tobramycin(10μl) were obtained from OXOID LTD., Basingstoke, Hampshire, England. Nalidixic acid(30μl), ciprofloxacin(5μl), trimethoprim/sulfamethoxazole (1.25/23.75μl), nitrofurantoin(300μl), and tetracycline(30μl) from Liofilchem, Roseto degli Abruzzi, Italy.
The following antimicrobial discs were used: ampicillin (10μl), amoxicillin/clavulanic acid (20/10μl), piperacillin/tazobactam (100/10μl), cefazolin (30μl), cefuroxime (30μl), ceftazidime (30μl) obtained from Hardy Diagnostics, Santa Maria, CA, USA. Ceftriaxone (30μl), cefotaxime (30μl), cefepime (30μl), imipenem (10μl), meropenem (10μl), amikacin (30μl), gentamicin (10μl), and tobramycin(10μl) were obtained from OXOID LTD., Basingstoke, Hampshire, England. Nalidixic acid(30μl), ciprofloxacin(5μl), trimethoprim/sulfamethoxazole (1.25/23.75μl), nitrofurantoin(300μl), and tetracycline(30μl) from Liofilchem, Roseto degli Abruzzi, Italy.
5
Automated Characterization of Bacterial Isolates
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Oxi/Ferm Pluri Test® (Liofilchem, Italy) was used. Next, the automatic characterization was carried out with the VITEK® 2 equipment and with the VITEK®2 GN identification cards (bioMérieux, Marcy-l’Étoile, France). The motility of the bacterium was tested in Motility Test Agar, (Liofilchem, Italy). Antimicrobial sensitivity was determined using E-test in Müller Hinton agar (Pronadisa®, Madrid, Spain) using the following antibiotics: piperacillin and piperacillin with tazobactam, cefepime (bioMérieux, Marcy-l’Étoile, France); ceftazidime, imipenem, imipenem with EDTA, amikacin, gentamicin, and ciprofloxacin (Liofilchem, Italy).
6
Multidrug-Resistant S. aureus Isolation
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S. aureus bacterial strains were isolated from 20 sewage samples collected from informal settlements, sewage treatment plants and abattoirs within Nairobi County, Kenya. Specimens from all the samples were streaked on selective Mannitol salt agar (Liofilchem®, Roseto degli Abruzzi, Italy) supplemented with 4 µg of ciprofloxacin [Liofilchem®, Roseto degli Abruzzi, Italy]. Single colonies were amplified in nutrient agar (HiMedia, Mumbai, India). Staphylococcus strains were identified using microscopy, physiological tests and an Analytical Profile Index of Staphylococcus system [bioMérieux, Marcy l’Etoile, France]. We screened isolated S. aureus bacteria for antimicrobial resistance to the following drugs: ceftazidime 30 µg, oxacillin 1 µg, vancomycin 30 µg, netilmicin 30 µg, gentamicin 10 µg, erythromycin 15 µg, trimethroprim-sulfamethoxazole 25 µg and cefuroxime 30 µg (Liofilchem®, Roseto degli Abruzzi, Italy), according to the Clinical Laboratory Standard Institute protocol.20 Isolates were considered to be MDRSA, if they were non-susceptible to more than one class of antibiotics. Colonies identified as MDRSA were re-suspended in 50% glycerol-nutrient broth and stored at -20 °C until use.
7
Campylobacter Antibiotic Susceptibility Testing
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Campylobacter isolates were subcultured on Campylobacter agar with 5% sheep blood (Liofilchem, Italy) and incubated for 48 h at 42 °C in a micro-aerophilic atmosphere. The suspension for each bacterial isolate at 0.5 McFarland turbidity was inoculated onto Mueller–Hinton agar with 5% sheep blood. Erythromycin (0.016–256 mg/L), tetracycline (0.016–256 mg/L), and ciprofloxacin (0.002–32 mg/L) strips (Liofilchem, Italy) were placed on these agar media, and the plates were incubated at 42 °C in a microaerophilic atmosphere for 24 h (12 (link)). The results were interpreted according to the Clinical and Laboratory Standards Institute (CLSI) guideline (13 ).
8
Antimicrobial Activity Evaluation Protocol
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Antimicrobial activity was assessed by depositing 10 μl of the crude extract and dilutions (MICs) on a 6-mm disc which was deposited on an inoculated Petri dish. Gentamicin and ciprofloxacin (Liofilchem) were used as a control following the Clinical and Laboratory Standards Institute method.[9 10 (link)]
9
Antibiotic Susceptibility of P. aeruginosa Morphotypes
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The antimicrobial susceptibility profile of each morphotype was determined using the disk diffusion method for ten antibiotics commonly used to treat P. aeruginosa infections in CF context: amikacin (AK) 30 μg, aztreonam (ATM) 30 μg, ciprofloxacin (CIP) 5 μg, tobramycin (TOB) 10 μg, gentamicin (CN) 10 μg, ceftazidime (CAZ) 30 μg, cefepime (FEP) 30 μg, imipenem (IMI) 10 μg, piperacillin (PRL) 100 μg and ticarcillin/clavulanic acid (TTC) 85 (75 + 10) μg, all obtained from Liofilchem. Disk diffusion susceptibility testing was performed in accordance with the recommendations of NCCLS document M100-522 (Clinical and Laboratory Standards Institute, 2012). The cartridges of antibiotic disks were stored at -20 °C and allowed to come to room temperature prior to use. The inhibition zones were measured (mm) after 18-21 h at 37 °C in air conditions. Colony morphotypes-associated bacteria were assigned as resistant (R), intermediate resistant (I) or sensitive (S) as defined by NCCLS guidelines in document M100-S28 (Clinical and Laboratory Standards Institute, 2018).
10
Antimicrobial Susceptibility Testing Protocol
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At first, using disk diffusion method, the susceptibility pattern of isolates was determined against gentamicin (120 µg), ciprofloxacin (5 µg) and vancomycin (30 µg) (Liofilchem Co, Roseto degli Abruzzi, Italy). Then, based on the results, the following concentration ranges were used for detection of MIC by broth microdilution method as recommended by clinical and laboratory standards institute [17 ]: gentamicin (15.625 – 8,000 µg/ml) and ciprofloxacin (1 – 512 µg/ml) (Exir pharmaceutical company, Borujerd, Lorestan, Iran).
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