The isolates positive for at least one virulence marker gene by PCR were subjected to antimicrobial susceptibility test against the selected antimicrobials (ampicillin-10 μg, amikacin-10 μg, chloramphenicol-30 μg, ceftriaxone-10 μg, cephalexin-30 μg, ciprofloxacin-10 μg, co-trimoxazole-25 μg, cefoperazone-tazobactam-75 + 10 μg, meropenem-10 μg, norfloxacin-10 μg, gentamicin-10 μg, cefixime-5 μg, doxycycline hydrochloride-10 μg and ofloxacin-5 μg) (HiMedia, India) by disc diffusion method in Mueller–Hinton agar [18 (link)]. The performance of this test was checked by employing E. coli ATCC 25922 as a standard quality control strain. The results were expressed as sensitive, intermediate and resistant as per standard CLSI guidelines [19 ]. MDR was defined as “acquired non-susceptibility to at least one agent in three or more antimicrobial categories” [20 (link)].
Norfloxacin
Manufactured by HiMedia
Sourced in India
Norfloxacin is a synthetic antibacterial agent that belongs to the quinolone class of antimicrobial drugs. It is used as a laboratory standard in research and development applications.
Automatically generated - may contain errors
Lab products found in correlation
Ciprofloxacin, by HiMedia (16 mentions)
Tetracycline, by HiMedia (14 mentions)
Ampicillin, by HiMedia (13 mentions)
Gentamicin, by HiMedia (12 mentions)
Ceftriaxone, by HiMedia (9 mentions)
Amikacin, by HiMedia (9 mentions)
Cefotaxime, by HiMedia (9 mentions)
Chloramphenicol, by HiMedia (8 mentions)
Meropenem, by HiMedia (7 mentions)
Erythromycin, by HiMedia (7 mentions)
Ceftazidime, by HiMedia (7 mentions)
Imipenem, by HiMedia (7 mentions)
Cotrimoxazole, by HiMedia (6 mentions)
Nalidixic acid, by HiMedia (6 mentions)
Azithromycin, by HiMedia (6 mentions)
Levofloxacin, by HiMedia (5 mentions)
Amoxicillin, by HiMedia (5 mentions)
Tobramycin, by HiMedia (5 mentions)
Cefoperazone, by HiMedia (4 mentions)
Cephalexin, by HiMedia (4 mentions)
20 protocols using norfloxacin
1
Antimicrobial Resistance Profiling of Virulent Isolates
2
Antibiotic Susceptibility Screening of Isolates
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Antibiotic drug susceptibility was determined by spreading overnight grown culture of the isolates on MRS agar plates as a lawn. Standard antibiotic discs (tetracycline, erythromycin, ampicillin, gentamycin, penicillin, chloramphenicol, cefuroxime, cefoperazone, levofloxacin, norfloxacin, Hi-Media, Mumbai) were placed on the surface of the MRS agar medium aseptically. Plates were incubated for 24 h at 37 °C and observed for zones of inhibition.
3
Antibiotic Resistance Profiling of E. coli
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The antibiotics resistance pattern of E. coli isolates against 20 different antibiotics such as ampicillin (10 mcg), amoxicillin (10 mcg), ceftazidime (30 mcg), cefoperazone (75 mcg), cefpodoxime (30 mcg), cefepime (30 mcg), clotrimazole (10 mcg), cefazolin (30 mcg), cefotaxime (30 mcg), cotrimaxozole (25 mcg), ceftriaxone (30 mcg), chloramphenicol (30 mcg), meropenem (10 mcg), moxifloxacin (5 mcg), nystatin (100 units), nitrofurantoin (300 mcg), norfloxacin (10 mcg), nitrofurantoin (300 mcg), penicillin (10 units) and tetracycline (30 mcg) (HiMedia, India) was performed on Mueller Hinton agar at 37 o C for 24 hrs as per standard agar-disc diffusion method (Bauer et al., 1966) . The selection of antimicrobial drugs was based on their common use, availability and as per the recommendations of Clinical and Laboratory Standards Institute (CLSI). After incubation, the zone of inhibition was measured and interpreted as sensitive, resistant and intermediate sensitive as per CLSI recommendations (Clinical and Laboratory Standards Institute, 2014).
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
Bacteriological Analysis and Antibiotic Susceptibility
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The swabs collected were directly inoculated in duplicate on blood agar, (Oxoid) Mac Conkey agar and Muller Hinton agar (Oxoid). The pairs of inoculated media were incubated aerobically at 37 °C for 24 h and then examined for bacteria growth according to standard protocol [14 ]. The positive culture growths were examined for colony characteristics, Gram-reaction and biochemical characteristics as described by Cheesbrough (2000) [14 ]. The biochemical characteristics tested were: (i) catalase, (ii) coagulase, (iii) oxidase, (iv) hemolysis, (v) sugar fermentation, (vi) indole production, (vii) citrate utilization, (viii) urease activity, (ix) triple sugar iron and (x) hydrogen sulphide production.
Purified bacterial isolates were subjected to drug susceptibility tests using the Kirby Bauer disc diffusion method [14 ]. Commercially available antibiotic discs that were used include: ciprofloxacin, norfloxacin, gentamycin, erythromycin, clindamycin, cephalexin, co-trimoxazole, tetracycline, cefoxitin, ampicillin, vancomycin and chloramphenicol (Hi Media Laboratories Pvt. Ltd, India). These antibiotics were chosen based on the type of microorganisms frequently isolated and their availability at University Teaching Hospital.
Purified bacterial isolates were subjected to drug susceptibility tests using the Kirby Bauer disc diffusion method [14 ]. Commercially available antibiotic discs that were used include: ciprofloxacin, norfloxacin, gentamycin, erythromycin, clindamycin, cephalexin, co-trimoxazole, tetracycline, cefoxitin, ampicillin, vancomycin and chloramphenicol (Hi Media Laboratories Pvt. Ltd, India). These antibiotics were chosen based on the type of microorganisms frequently isolated and their availability at University Teaching Hospital.
6
Antimicrobial Resistance Profiling of MRSA
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Resistance to antibiotics was confirmed as per the guidelines of internationally recognized standards of the Clinical and Laboratory Standards Institute (CLSI)53 . MRSA, MRSE and MRSH strains were screened for susceptibility by 22 antimicrobial agents using disc diffusion method on Muller-Hinton agar. The antibiotics disc tested were (concentration in μg, HiMedia, India) Erythromycin (E-15 μg), Clindamycin (CD-2 μg), Gentamicin (GEN-10 μg), Cefotaxime (CTX-30 μg), Norfloxacin (NX-10 μg), Ciprofloxacin (CIP-5 μg), Tetracycline (TE-30 μg), Rifampicin (RIF-5 μg), Nitrofurantoin (NIT-300 μg), Linezolid (LZ-30 μg), Chloramphenicol (C-30 μg), Amoxyclav (AMC-10 μg), Amphicillin (AMP-10 μg), Azithromycin (AZM-15 μg), Bacitracin (B-10 Units), Cefoxitime (CFM-5 μg), co-Trimoxazole (COT-23.75 μg), Kanamycin (K- 30 μg), Oxacillin (OX-I μg), Penicillin G (P-10 Units), Streptomycin (S-100 μg), Tetracycline (TE-30 μg) and Tobramycin (TOB-30 μg). The results were recorded after 24 h of incubation at 37 °C and interpreted as per CLSI guidelines. Screened isolates showing resistance to more than three classes of non-β-lactam antibiotics were considered as multidrug resistant (MDR) strains, as described by Chajecka et al.,9 (link). Assay was performed in triplicates for all strains.
7
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.
8
Enzymatic Assay for Cysteine Sulfoxide Synthesis
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Reduced
form of nicotinamide adenine dinucleotide
(NADH), lactate dehydrogenase (LDH) from rabbit muscle, and (±)-L-alliin
were purchased from Sigma-Aldrich; pyridoxal 5′-phosphate (PLP)
andd ,l -dithiothreitol (DTT) were from Serva; kanamycin
is a domestic product (OAO Biokhimik); DEAE-sepharose was from Amersham.
2-Nitro-5-thiobenzoate (NTB) was prepared according to ref (31 (link)). S-Methyl-l -cysteine sulfoxide (methiin) was synthesized according to
Morozova et al.22 (link) PEG–poly(α,β-aspartic
acid)70 (PEG–P(Asp)70) and poly-(l -lysine)70 (PLL70) were synthesized according
to Koide et al.32 (link) Luria–Bertani
broth (LB), Mueller–Hinton broth, Mueller–Hinton agar,
and antibiotic-impregnated discs: amikacin, amoxycillin/clavulanic
acid, ampicillin, azithromycin, aztreonam, cefepime, ceftazidime,
ceftriaxone, cephotaxime, cephoxitin, chloramphenicol, ciprofloxacin,
colistin, doxycycline, erythromycin, gentamicin, imipenem, levofloxacin,
lincomycin, norfloxacin, ofloxacin, oxacillin, rifampicin, spiramycin,
tobramycin, and vancomycin were from HiMedia Laboratories Pvt. Limited
(India).
form of nicotinamide adenine dinucleotide
(NADH), lactate dehydrogenase (LDH) from rabbit muscle, and (±)-L-alliin
were purchased from Sigma-Aldrich; pyridoxal 5′-phosphate (PLP)
and
is a domestic product (OAO Biokhimik); DEAE-sepharose was from Amersham.
2-Nitro-5-thiobenzoate (NTB) was prepared according to ref (31 (link)). S-Methyl-
Morozova et al.22 (link) PEG–poly(α,β-aspartic
acid)70 (PEG–P(Asp)70) and poly-(
to Koide et al.32 (link) Luria–Bertani
broth (LB), Mueller–Hinton broth, Mueller–Hinton agar,
and antibiotic-impregnated discs: amikacin, amoxycillin/clavulanic
acid, ampicillin, azithromycin, aztreonam, cefepime, ceftazidime,
ceftriaxone, cephotaxime, cephoxitin, chloramphenicol, ciprofloxacin,
colistin, doxycycline, erythromycin, gentamicin, imipenem, levofloxacin,
lincomycin, norfloxacin, ofloxacin, oxacillin, rifampicin, spiramycin,
tobramycin, and vancomycin were from HiMedia Laboratories Pvt. Limited
(India).
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
Antibiotic Susceptibility Profiling of A. hydrophila
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Antibiotic susceptibility profile for A. hydrophila Ah17 was determined by the Kirby‐Bauer disk diffusion method (Bauer, Kirby, Sherris, & Turck, 1966 ). The following antibiotics were tested: Amikacin (AK: 30 μg), Amoxicillin (AMC: 30 μg), Ampicillin (AMP: 10 μg), Azithromycin (AZM: 15 μg), Cefixime (CFM: 5 μg), Cefoperazone (CPZ: 75 μg), Cefpodoxime (CPD: 10 μg), Ciprofloxacin (CIP: 5 μg), Chlorompenicol (C: 30 μg), Clarithromycin (CLR: 15 μg), Co‐Trimoxazole (COT: 25 μg), Doxycycline hydrochloride (DO: 30 μg), Fosfomycin (FO: 200 μg), Fusidic acid (FC: 10 μg), Gentamicin (GEN: 10 μg), Imipenem (IPM: 10 μg), Kanamycin (K: 30 μg), Linezolid (LZ: 30 μg), Methicillin (MET: 5 μg), Minocycline (MI: 30 μg), Nalidixic acid (NA: 30 μg), Nitrofurantoin (NIT: 300 μg), Norfloxacin (NX: 10 μg), Penicillin G (P: 10 μg), Pristinomycin (RP: 15 μg), Rifampicin (RIF: 5 μg), Streptomycin (S: 10 μg), Teicoplanin (TEI: 30 μg), Tetracycline (TE: 30 μg), Trimethoprim (TR: 5 μg), Tobramycin (TOB: 10 μg), and Vancomycin (VA: 30 μg) (HiMedia). 200 μl of A. hydrophila Ah17 suspension was swabbed on Mueller‐Hinton agar (MHA) medium (HiMedia) and kept for drying. Antibiotic discs were placed on the MHA medium and incubated at 37°C for 24–48 hr. The diameter of the zone of inhibition was measured, and susceptibility was categorized according to the manufacturer's protocol.
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