Antimicrobial susceptibility of the isolates to the following antibiotics were carried out by the Kirby-Bauer (KB) disk diffusion method according to the Clinical Laboratory Standards Institute (CLSI 2012) guide lines (19 ); tetracycline (TE) [30 μg], amikacin (AN) [30 μg], tigecycline (TIG) [15 μg], colistin (CL) [10 μg], gentamicin (GM) [10 μg], piperacillin (PIP) [100 μg], ciprofloxacin (CIP) [5 μg], ceftazidime (CAZ) [30 μg], cefotaxime (CTX) [30 μg], tobramycin (TOB) [10 μg], amoxicillin + clavulanic acid (AMC) [30 μg], rifampin (Rif) [30 μg], cefixime (CFM) [5 μg], nalidixic acid (NA) [30 μg], imipenem (IMP) [10 μg] and meropenem (MEM) [10 μg] (Hi-media-India). Susceptibility to tigecycline was classified based on EUCAST criteria (20 (link)) (MIC ≤ 0.5 μg/mL; inhibition zone ≤ 17 mm). Minimum inhibitory concentrations (MICs) of the IMP and MEM were determined by E-test (Hi-Media, India) as described by the manufacturer’s instructions and interpreted according to CLSI guidelines (19 ). E. coli ATCC 25922 was used as the quality control strain.
Nalidixic acid
Manufactured by HiMedia
Sourced in India
Nalidixic acid is a synthetic quinolone antibiotic used in the laboratory setting. It functions as a bactericidal agent by inhibiting DNA gyrase, an essential enzyme for bacterial DNA replication and transcription.
Automatically generated - may contain errors
Lab products found in correlation
Ciprofloxacin, by HiMedia (23 mentions)
Tetracycline, by HiMedia (17 mentions)
Cotrimoxazole, by HiMedia (17 mentions)
Ampicillin, by HiMedia (17 mentions)
Chloramphenicol, by HiMedia (15 mentions)
Ceftriaxone, by HiMedia (13 mentions)
Ceftazidime, by HiMedia (12 mentions)
Cefotaxime, by HiMedia (11 mentions)
Gentamicin, by HiMedia (10 mentions)
Imipenem, by HiMedia (9 mentions)
Cefixime, by HiMedia (8 mentions)
Azithromycin, by HiMedia (8 mentions)
Erythromycin, by HiMedia (7 mentions)
Streptomycin, by HiMedia (7 mentions)
Amikacin, by HiMedia (6 mentions)
Norfloxacin, by HiMedia (6 mentions)
Amoxicillin, by HiMedia (6 mentions)
Gentamycin, by HiMedia (6 mentions)
Levofloxacin, by HiMedia (5 mentions)
Colistin, by HiMedia (5 mentions)
35 protocols using nalidixic acid
1
Antimicrobial Susceptibility Testing of Isolates
2
MIC Variation Under Gut Conditions
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MIC was determined to check the effect of infection-related in vitro gut conditions, such as bile, osmotic, high and low iron, pH and temperature conditions, on MIC variation in resistant and sensitive isolate by using Ezy MIC™ strips of ampicillin, co-trimoxazole, imipenem, nalidixic acid, ciprofloxacin, tetracycline, nitrofurantoin and chloramphenicol (HiMedia Laboratories Pvt. Ltd., Maharashtra, India). Both the isolates were grown up to mid-exponential phase (MEP) under in vitro gut conditions and swabbed onto Muller Hinton agar (MHA) plates. MIC strips were placed onto plates using an applicator followed by incubation at 37 °C for 16–18 h. The result was read where the ellipse intersects the MIC scale on the strip for the tested antibiotics.
3
Antimicrobial Resistance Profile of Campylobacter
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The AMR profile of Campylobacter isolates was determined using standard Kirby-Bauer disc diffusion method as described by Taremi et al. [16 (link)]. A total of 38 revived isolates were tested against a panel of eight antibiotics that included ampicillin (AMP, 10 µg), gentamicin (GEN, 10 µg), ERY (15 µg), levofloxacin (LE, 5 µg), CIP (5 µg), nalidixic acid (NA, 30 µg), ceftriaxone (CTR, 30 µg), and co-trimoxazole (COT, 25 µg) (HiMedia). The isolates were revived on mCCDA plates supplemented with FD009 supplement. The growth suspension prepared in Tryptic soy broth and compared with 0.5 McFarland standard was spread on Mueller-Hinton agar plates supplemented with 7% sheep blood and incubated at 42°C in a CO2 incubator at 5% CO2 tension for 24 h. Zone diameter was measured and breakpoints were interpreted based on the recommendations of the Clinical and Laboratory Standards Institute standards for disc diffusion assay (CLSI 2016).
4
Antimicrobial Susceptibility Profiling
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A total of 18 clinically relevant antibiotics were tested using the disc diffusion method (Kirby-Bauer's) and the inhibition zone diameters were measured by (mm) according to Clinical and Laboratory Standards Institute (CLSI) guidelines M100 27 th 18 . These antimicrobial discs were amoxicillin/clavulanic acid ( 30μg), ampicillin (10 μg), cefotaxime (30 μg), ceftazidime (30 μg), ceftriaxone (30 μg), aztreonam (30 μg), imipenem (10 μg), meropenem (10 μg), ertapenem(10 μg), gentamicin (10 μg), tobramycin (10 μg) , amikacin (30 μg), tetracycline (30 μg), ciprofloxacin (5 μg), norfloxacin (10 μg), nalidixic acid (30 μg), co-trimoxazole (25 μg) and colistin (10 μg) were obtained from HiMedia Laboratories (India). The results for the antimicrobial susceptibility test strain were interpreted as (S) susceptible, (I) intermediate, or (R) resistant by comparing the results to the CLSI 2017 18 standard zone diameter Quality control strains used in antimicrobial susceptibility testing are Escherichia coli ATCC #25922 18 . Minimum inhibitory concentration (MIC) of imipenem and meropenem were determined using the agar dilution method and interpreted according to the CLSI guidelines, the carbapenems resistant Enterobacteriaceae (CRE) isolates were included based on showing MICs ≥2 µg/mL for imipenem or meropenem were considered resistant. Only one isolate per patient was included 19 .
5
Antibiotic Susceptibility of Salmonella
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Antimicrobial susceptibility testing of Salmonella isolates was performed by using the modified Kirby-Bauer disk diffusion method as recommended by CLSI guidelines. The antibiotics used in this study were ampicillin (AMP, 10 μg), azithromycin (AZM, 15 μg), cefixime (CFM, 5 μg), cefotaxime (CTX, 30 μg), ceftriaxone (CTR, 30 μg), ciprofloxacin (CIP, 5 μg), chloramphenicol (C, 30 μg), cotrimoxazole (COT, 25 μg), levofloxacin (LEV, 5 μg), nalidixic acid (NA, 30 μg) and ofloxacin (OF, 5 μg) from HiMedia [17 ]. MDR Salmonella species were identified based on the resistance to the three first-line drugs [18 (link)].
6
Antimicrobial Susceptibility Testing of Escherichia coli
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This was performed with the disk diffusion method (Kirby-Bauer’s) and the inhibition zone diameters were measured by (mm) according to Clinical and Laboratory Standards Institute (CLSI).12 (link) The antimicrobial discs tested include Amoxicillin/clavulanic acid (30 μg), ampicillin (10 μg), cefotaxime (30 μg), ceftazidime (30 μg), ceftriaxone (30 μg), aztreonam (30 μg), imipenem (10 μg), meropenem (10 μg), ertapenem (10 μg), gentamicin (10 μg), tobramycin (10 μg), amikacin (30 μg), tetracycline (30 μg), ciprofloxacin (5 μg), norfloxacin (10 μg), nalidixic acid (30 μg), co-trimoxazole (25 μg) and colistin (10 μg) were obtained from HiMedia Laboratories (India). The results for the antimicrobial susceptibility test strain were interpreted as (S) susceptible, (I) intermediate or (R) resistant by comparing the results to the CLSI 2018 standard zone diameter.12 (link) Minimum inhibitory concentrations (MICs) for imipenem and meropenem were performed by the agar dilution method and interpreted according to CLSI guidelines (strains displaying MICs ≥8 µg/mL for imipenem and meropenem were considered resistant).12 (link)
Escherichia coli ATCC 25922 was used as standard control strains.
Escherichia coli ATCC 25922 was used as standard control strains.
7
Antimicrobial Susceptibility Testing of Bacterial Isolates
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Antimicrobial susceptibility testing of the isolated strains was performed using the disk diffusion method (modified Kirby–Bauer method) on Mueller–Hinton agar (HiMedia, India) as recommended by the CLSI, Wayne, United States.
4
Susceptibility of the fluoroquinolones including nalidixic acid (30 µg), pefloxacin (5 µg), ciprofloxacin (5 µg), azithromycin (15 µg), chloramphenicol (30 µg), cotrimoxazole(1.25 µg/23.75 µg), cefixime (5 µg), and ceftriaxone (30 µg) (HiMedia Laboratories, India) was done. The results of the antibiotic susceptibility were determined on the basis of interpretative zone diameters as suggested by CLSI. For standardization,
Escherichia coliATCC-25922 was used as the control organism for antibiotic sensitivity.
Further, MICs of all isolates was checked using broth microdilution method as per CLSI guidelines.
7
ciprofloxacin concentrations ranged from 0.06 to 16 µg/mL. pefloxacin HiComb (HiMedia),was used for determining the MIC of pefloxacin, which is available as Part A and Part B with concentration of 240 to 0.01 µg/mL and 30 to 0.001 µg/mL, respectively. Antimicrobial susceptibility testing was performed according to the manufacturer’s instructions and interpreted using CLSI guidelines.
E. coliATCC 25922 and
Pseudomonas aeruginosaATCC 27853 served as quality control strains.
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Antimicrobial susceptibility testing of the isolated strains was performed using the disk diffusion method (modified Kirby–Bauer method) on Mueller–Hinton agar (HiMedia, India) as recommended by the CLSI, Wayne, United States.
4
Susceptibility of the fluoroquinolones including nalidixic acid (30 µg), pefloxacin (5 µg), ciprofloxacin (5 µg), azithromycin (15 µg), chloramphenicol (30 µg), cotrimoxazole(1.25 µg/23.75 µg), cefixime (5 µg), and ceftriaxone (30 µg) (HiMedia Laboratories, India) was done. The results of the antibiotic susceptibility were determined on the basis of interpretative zone diameters as suggested by CLSI. For standardization,
Escherichia coliATCC-25922 was used as the control organism for antibiotic sensitivity.
Further, MICs of all isolates was checked using broth microdilution method as per CLSI guidelines.
7
ciprofloxacin concentrations ranged from 0.06 to 16 µg/mL. pefloxacin HiComb (HiMedia),was used for determining the MIC of pefloxacin, which is available as Part A and Part B with concentration of 240 to 0.01 µg/mL and 30 to 0.001 µg/mL, respectively. Antimicrobial susceptibility testing was performed according to the manufacturer’s instructions and interpreted using CLSI guidelines.
E. coliATCC 25922 and
Pseudomonas aeruginosaATCC 27853 served as quality control strains.
8
Antibiotic Susceptibility Testing of Bacterial Isolates
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The antimicrobial susceptibility of bacterial isolates was performed by using Kirby Bauer Disk diffusion method on Mueller Hinton Agar (Hi-Media, India) according to the CLSI guidelines [15 ]. The antibiotic susceptibility pattern was examined by using commercial antibiotic discs including Amoxicillin (10μg), Ceftazidime (30μg), Gentamicin (10μg), Tetracycline (30μg), Nitrofurantoin (300μg), Cotrimoxazole (1.25μg), Nalidixic acid (30μg), Cefoxitin (30μg), Azithromycin (15μg), and Ciprofloxacin (5μg) (Hi-Media, India). The Escherichia coli isolate ATCC 25922 and Staphylococcus aureus isolate ATCC 25923 were used as reference organisms for quality control to antimicrobial susceptibility testing.
9
Antimicrobial Susceptibility of Pathogenic Vibrio
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Antimicrobial susceptibility testing of the pathogenic (tdh+) V. parahaemolyticus isolates were performed using the disk diffusion method with commercially available antibiotic disks such as ampicillin (10 µg), cefotaxime (30 µg), cefpodoxime (10 µg), ceftazidime (30 µg), ceftizoxime (30 µg), ceftriaxone (30 µg), chloramphenicol (30 µg), ciprofloxacin (5 µg), gentamicin (10 µg), levofloxacin (5 µg), nalidixic acid (30 µg), norfloxacin (10 µg), tetracycline (30 µg) and trimethoprim (5 µg) (HiMedia, India) in accordance with the criteria recommended by Clinical and Laboratory Standards Institute (CLSI 2010 ). Escherichia coli ATCC 25,922 was used as a control strain.
10
Antimicrobial Susceptibility Testing of E. coli
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Antimicrobial susceptibility testing was performed via a standard disk diffusion method, also known as the Kirby–Bauer method, according to the protocols of the CLSI [36 ]. We again used antibiotics applicable to the treatment of patients, namely meropenem (10 µg, MEM10C Oxoid ltd, Basingstoke, Hampshire, UK), ampicillin (10 µg, SD002-1PK), amoxycillin (25 µg, SD129-1PK), amoxycillin/clavulanic acid (20/10 µg, AUG30C), carbenicillin (100 µg, SD004-1PK), cefamandole (30 µg, SD200-1PK), erythromycin (15 µg, SD013-1PK), streptomycin (10 µg, SD031-1PK), tetracycline (30 µg, SD037-1PK), doxycycline hydrochloride (30 µg, SD012-1PK), chloramphenicol (30 µg, SD006-1PK), nalidixic acid (30 µg, SD021-1PK), ciprofloxacin (5 µg, SD060-1PK), pefloxacin (5 µg, SD070-1PK) and co-trimoxazole (25 µg, SD010-1PK) from HiMedia, India. The results were evaluated according to the cut-off breakpoint values of EUCAST version 12.0, 2022 [37 ], CLSI, 31st edition [36 ] and the Manual of BBL Products and Laboratory Procedures [38 ]. Breakpoint values of erythromycin for other bacterial species were taken for E. coli.
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