Antimicrobial susceptibility was determined by Kirby–Bauer disc diffusion method13 (link) on MHA plates. The following antibiotics were used: amikacin (30 μg), gentamicin (10 μg), ciprofloxacin (30 μg), trimethoprim/sulphamethoxazole (1.25/23.75 μg), cefepime (30 μg), imipenem (10 μg), meropenem (10 μg), ceftriaxone (30 μg) and aztreonam (30 μg) (HiMedia). Minimum inhibitory concentrations (MICs) of various antibiotics were determined on MHA plates by agar dilution method according to CLSI guidelines14 using the following antibiotics: cefotaxime, ceftazidime, ceftriaxone, cefepime, imipenem, meropenem, ertapenem and aztreonam (HiMedia).
Aztreonam
Manufactured by HiMedia
Sourced in India
Aztreonam is a synthetic antibacterial agent that belongs to the class of monobactam antibiotics. It is a semisynthetic beta-lactam antibiotic used to treat Gram-negative bacterial infections.
Automatically generated - may contain errors
Lab products found in correlation
Ciprofloxacin, by HiMedia (20 mentions)
Ceftazidime, by HiMedia (20 mentions)
Imipenem, by HiMedia (19 mentions)
Gentamicin, by HiMedia (17 mentions)
Amikacin, by HiMedia (15 mentions)
Cefotaxime, by HiMedia (15 mentions)
Meropenem, by HiMedia (13 mentions)
Cefepime, by HiMedia (13 mentions)
Ampicillin, by HiMedia (13 mentions)
Ceftriaxone, by HiMedia (12 mentions)
Ofloxacin, by HiMedia (8 mentions)
Levofloxacin, by HiMedia (8 mentions)
Piperacillin tazobactam, by HiMedia (7 mentions)
Tobramycin, by HiMedia (7 mentions)
Ertapenem, by HiMedia (6 mentions)
Colistin, by HiMedia (6 mentions)
Amoxicillin clavulanic acid, by HiMedia (5 mentions)
Cotrimoxazole, by HiMedia (5 mentions)
Piperacillin, by HiMedia (4 mentions)
Tetracycline, by HiMedia (4 mentions)
21 protocols using aztreonam
1
Antibiotic Susceptibility Testing Protocol
2
Antibiotic Susceptibility Testing of Isolates
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Antibiotic susceptibility tests of all isolates were performed using Kirby Bauer disc diffusion method on Mueller-Hinton Agar with recommended antibiotics by CLSI 2020 guidelines [14 ].The antibiotics used were gentamicin (GEN,30 µg), amikacin (AK, 10 µg), ciprofloxacin (CIP, 5 µg), ceftazidime (CAZ, 30 µg), cefepime (CPM, 30 µg), aztreonam (AT, 30 µg), imipenem (IPM, 10 µg), piperacillin (PI,30 µg), piperacillin-tazobactam (PIT), meropenem (MRP, 10 µg), ofloxacin (OF, 30 µg), Levofloxacin (LEV, 30 µg) and colistin (CL,10 µg) from Hi-Media, Laboratories Pvt. Ltd. India.
Isolates that were non-susceptible to at least one agent in ≥ 3 antimicrobial categories have been categorized under MDR [15 (link)].
Isolates that were non-susceptible to at least one agent in ≥ 3 antimicrobial categories have been categorized under MDR [15 (link)].
3
Antibiotic Resistance Profiling of Clinical Isolates
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Antibiotic susceptibility test (AST) of both clinical isolates was performed using the modified Kirby-Bauer disk diffusion method on Mueller Hinton agar (Hi-Media Laboratories, India) following standard zone size interpretative criteria set by the Clinical and Laboratory Standards Institute (CLSI) [25 ]. The different antibiotic disks used in this study during AST were procured from HiMedia Laboratories, India, and include amoxicillin (30 μg), gentamicin (10 μg), cotrimoxazole (25 μg), ciprofloxacin (5 μg), imipenem (10 μg), amoxicillin/clavulanic acid (20/10 μg), cefotaxime (30 μg), ceftriaxone (30 μg), ceftazidime (30 μg), aztreonam (30 μg), and cefpodoxime (10 μg). The E. coli and K. pneumoniae isolates were regarded as MDR isolates if they were resistant to at least one agent of three different classes of antimicrobial disks [2 (link)].
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 Testing Protocol
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Antibiotic susceptibility testing (AST) was performed following the modified Kirby Bauer disc diffusion method using CLSI guidelines (2016) as a reference [17 ]. A total of 13 different commonly prescribed antibiotics (tetracycline (30 µg), imipenem (10 µg), chloramphenicol (30 µg), ciprofloxacin (5 µg), gentamycin (10 µg), azithromycin (15 µg), methicillin (5 µg), ceftazidime (30 µg), cefotaxime (30 µg), cefepime (30 µg), amikacin (30 µg), aztreonam (30 µg), and levofloxacin (5 µg)) procured from Hi-Media, India, were used for susceptibility testing.
6
Antibiotic Susceptibility Profiling of Bacterial Isolates
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The susceptibility of bacterial isolates against different antibiotics was determined by the disk diffusion method [modified Kirby-Bauer method] on Mueller Hinton agar (Hi-Media, India) following standard procedures recommended by the Clinical and Laboratory Standards Institute (CLSI), Wayne, USA [18 ]. For this purpose following antibiotics with specified concentrations were used; ampicillin (10 μg), ampicillin-sulbactam (10/10 μg), ceftazidime (30 μg), ceftriaxone (30 μg), cefepime (30 μg), cefoxitin (30 μg), piperacillin-tazobactam (100/10 μg, aztreonam (30 μg), imipenem (10 μg), meropenem (10 μg), gentamycin (10 μg), amikacin (30 μg), ciprofloxacin (5 μg), ofloxacin (5 μg), levofloxacin (5 μg), trimethoprim-sulphamethoxazole/co-trimoxazole (25 μg), polymixin B (300unit), colistin sulphate (10 μg)] and tigecycline (30 μg) from HiMedia Laboratories, India. Interpretations of antibiotic susceptibility results were made according to the guidelines of interpretative zone diameters of CLSI [18 ]. Escherichia coli ATCC 25922 and Pseudomonas aeruginosa ATCC 27853 were used as the control organisms for antibiotic sensitivity.
7
ESBL Detection in E. coli Isolates
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As per CLSI protocol, E. coli isolates with a zone of inhibition of ≤17 mm for aztreonam and ceftazidime, and ≤22 mm for cefotaxime in disc diffusion susceptibility testing were selected for detection of ESBLs production. For this purpose, four antibiotics cefotaxime (10 μg), ceftazidime (30 μg), aztreonam (30 μg), and cefotaxime+clavulanic acid (30+10 μg) (HiMedia, India) were used. Discs were placed on the inoculated MHA plates at a distance of 25 mm apart and incubated overnight. The E. coli isolates resistant to either of the cephalosporin discs and sensitive to their respective cephalosporin+clavulanic acid discs with diameter of more than 5 mm were considered as presumptive ESBL producers [13 (link)].
8
Antimicrobial Susceptibility of E. coli
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All biochemically confirmed E. coli isolates were tested for their antimicrobial drug susceptibility pattern on Mueller-Hinton agar (MHA) (HiMedia, India) by the disc diffusion method [12 ]. The antibiotics used were oxytetracycline (30 μg), cephalexin (30 μg), ciprofloxacin (5 μg), gentamicin (30 μg), cefotaxime (10 μg), ampicillin (10 μg), ceftazidime (30 μg), aztreonam (30 μg), imipenem (10 μg), cefixime (5 μg), and meropenem (10 μg) (HiMedia, India). The diameter of the zones of complete inhibition was measured and compared with the zone size interpretation chart and was graded as sensitive, intermediate, and resistant. The MAR Index was also calculated for all E. coli isolates, by applying formula a/b where “a” is the number of antibiotics to which an isolate was resistant and “b” is the number of antibiotics to which the isolates exposed [13 (link)].
9
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.
10
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).
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