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).
Nitrofurantoin
Manufactured by HiMedia
Sourced in India
Nitrofurantoin is a laboratory reagent used in various research and analytical applications. It is a synthetic antimicrobial agent that has been widely studied for its functional properties. The core function of Nitrofurantoin is to serve as a chemical compound for use in scientific experiments and investigations, without further interpretation or extrapolation on its intended use.
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Lab products found in correlation
Ciprofloxacin, by HiMedia (14 mentions)
Ceftazidime, by HiMedia (12 mentions)
Amikacin, by HiMedia (11 mentions)
Ampicillin, by HiMedia (10 mentions)
Ceftriaxone, by HiMedia (9 mentions)
Gentamicin, by HiMedia (9 mentions)
Imipenem, by HiMedia (9 mentions)
Meropenem, by HiMedia (9 mentions)
Cefotaxime, by HiMedia (9 mentions)
Cotrimoxazole, by HiMedia (9 mentions)
Tetracycline, by HiMedia (6 mentions)
Cefepime, by HiMedia (6 mentions)
Piperacillin tazobactam, by HiMedia (6 mentions)
Cefoxitin, by HiMedia (6 mentions)
Cefixime, by HiMedia (4 mentions)
Nalidixic acid, by HiMedia (4 mentions)
Norfloxacin, by HiMedia (4 mentions)
Mueller hinton agar (mha), by HiMedia (4 mentions)
Chloramphenicol, by HiMedia (3 mentions)
Cefpodoxime, by HiMedia (3 mentions)
20 protocols using nitrofurantoin
1
Antibiotic Resistance Profiling of E. coli
2
Antibiotic Susceptibility Testing of Citrobacter freundii
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According to the Clinical and Laboratory Standards Institute [55 ], the AST of isolated C. freundii was done by a Kirby–Bauer disk diffusion test [56 (link)]. In this study, 14 antibiotics from ten classes were chosen based on their availability in Bangladesh: fluoroquinolones (ciprofloxacin—5 μg, levofloxacin—5 μg), aminoglycosides (gentamicin—10 μg), tetracyclines (tetracycline—30 μg), macrolides (azithromycin—15 μg), cephalosporins (ceftriaxone—30 μg, cephalexin—30 μg, cefotaxime—30 μg, ceftazidime—30 μg), penicillins (ampicillin—10 μg), glycopeptides (chloramphenicol—30 μg), sulfonamides (cotrimoxazole- 25 μg), phosphonic acids (fosfomycin—200 μg), nitrofurantoin (nitrofurantoin—100 μg) (HiMedia, India). A multidrug-resistant (MDR) isolate was characterized as one that is resistant to three or more antibiotic classes [57 (link)]. We enumerated the multiple antibiotic resistance (MAR) index by the following formula [47 (link)]: MAR = w/v; here, w = number of antibiotics to which an isolate is resistant, v = total number of antibiotics used in this study.
3
Antimicrobial Susceptibility Testing Protocol
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Antimicrobial susceptibility testing was performed using the Kirby-Bauer disk diffusion method, adhering to Clinical and Laboratory Standards Institute (CLSI) guidelines [26 ]. The selection of antibacterial disks was based on CLSI 2022 recommendations and availability. The bacterial inoculum, equivalent to 0.5 McFarland standards, was uniformly spread onto Mueller–Hinton agar plates (Himedia India) using a sterile cotton swab applicator. Antibiotic disks for gram-positive bacteria were penicillin G (10 μg), nitrofurantoin (300 μg), ciprofloxacin (5μg), trimethoprim-sulfamethoxazole (1.25/23.75 μg), gentamicin (10 μg), cefoxitin (30 μg), and clindamycin (30 μg) (Himedia, India).
The antimicrobial disks for Enterobacteriaceae included piperacillin-tazobactam (100/10 μg), ampicillin (10 μg), nitrofurantoin (300 μg), amoxicillin-clavulanate (20/10 μg), gentamicin (10 μg), cefotaxime (30 μg), ciprofloxacin (30 μg), meropenem (10 μg), piperacillin (100 μg), cefepime (30 μg), amikacin (30 μg), trimethoprim-sulfamethoxazole (1.25/23.75 μg), ceftazidime (30 μg), and ceftriaxone (30 μg) (Himedia, India). Antimicrobial disks for Pseudomonas spp and Acinetobacter were gentamicin (10 μg), meropenem (10 μg), ceftazidime (30 μg), piperacillin-tazobactam (100/10 μg), ciprofloxacin (5 μg), and amikacin (30 μg) (Himedia, India).
The antimicrobial disks for Enterobacteriaceae included piperacillin-tazobactam (100/10 μg), ampicillin (10 μg), nitrofurantoin (300 μg), amoxicillin-clavulanate (20/10 μg), gentamicin (10 μg), cefotaxime (30 μg), ciprofloxacin (30 μg), meropenem (10 μg), piperacillin (100 μg), cefepime (30 μg), amikacin (30 μg), trimethoprim-sulfamethoxazole (1.25/23.75 μg), ceftazidime (30 μg), and ceftriaxone (30 μg) (Himedia, India). Antimicrobial disks for Pseudomonas spp and Acinetobacter were gentamicin (10 μg), meropenem (10 μg), ceftazidime (30 μg), piperacillin-tazobactam (100/10 μg), ciprofloxacin (5 μg), and amikacin (30 μg) (Himedia, India).
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Enterococcus Antibiotic Susceptibility Patterns
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The present study was conducted at Vinayaka Mission's Medical College and Hospitals, Salem Institutional Ethical Committee (IEC) Clearance (MICROBIOLOGY/Ph.D/01/2015) was obtained prior to this study A total of 1200 clinical samples received at the Department of Microbiology for routine culture and sensitivity were included. A total of 774 isolates of Enterococci obtained from 1200 clinical samples were processed for species identification and antimicrobial sensitivity testing. Identification of isolates to the species level was performed by the sugar utilization test using brain heart infusion broth incorporated with 1% sugar and bromothymol blue indicator.[9 (link)] Isolates of Enterococci were subjected to antimicrobial susceptibility testing by the Kirby–Bauer disc diffusion method as per Clinical and Laboratory Standards Institute guidelines[10 ] using the following drugs: penicillin (10 U), erythromycin (15 μg), high-level gentamicin (120 μg), linezolid (30 μg), teicoplanin (30 μg), ciprofloxacin (5 μg), and nitrofurantoin (300 μg).(Himedia Laboratories Ltd.) The minimum inhibitory concentrations (MICs) for these antibiotics were also determined bythe Vitek automated system.
5
Antimicrobial Susceptibility Testing
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Amikacin (AMK; 30 μg/disk), ampicillin/sulbactam (SAM, 10/10 µg/disc), cefixime (CFM; 5 µg/disc), cefotaxime (CTX; 30 µg/disc), ceftazidime (CAZ; 30 µg/disc), ceftriaxone (CRO; 30 µg/disc), ciprofloxacin (CIP, 5 µg/disc), gentamicin (GEN, 10 µg/disc), imipenem (IPM, 10 µg/disc), meropenem (MEM; 10 µg/disc), nitrofurantoin (NIT; 300 µg/disc), sulbactam (SUL; 10 µg/disc), trimethoprim/sulfamethoxazole (SXT; 25 µg/ disc) (HiMedia Laboratories, Mumbai,. India), imipenem and sulbactam powder (Sigma-Aldrich Co. St. Louis, MO, USA). The antifungal powders were dissolved in dimethyl sulfoxide (DMSO) and stock solutions diluted based on Clinical and Laboratory Standard Institute (CLSI) guidelines (CLSI M07-A10).
6
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.
7
Antibiotic Susceptibility of Bacterial Isolates
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The susceptibility to the commercial antibiotics of the bacterial isolates was evaluated using the disc diffusion method. Antibiotics used against Gram-positive bacteria included cefoxitin, benzyl-penicillin, oxacillin, imipenem, gentamicin, ciprofloxacin, moxifloxacin, inducible clindamycin resistance, erythromycin, clindamycin, vancomycin, tetracycline, fusidic acid, and trimethoprim/sulfamethoxazole. On the other hand, antibiotics used against Gram-negative bacteria included temocillin, ampicillin, amoxicillin/clavulanic acid, ticarcillin, ticarcillin/clavulanic acid, piperacillin, piperacillin/tazobactam, cephalothin, cefuroxime, cefotaxime, ceftazidime, ceftriaxone, cefepime, ertapenem, imipenem, meropenem, amikacin, gentamicin, tobramycin, ciprofloxacin, tigecycline, fosfomycin, nitrofurantoin, pefloxacin, minocycline, colistin, and trimethoprim/sulfamethoxazole (Himedia Labs, Mumbai, India) [19 ].
8
Antimicrobial Susceptibility Testing Protocol
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The susceptibility of bacterial isolates against different antibiotics was tested by the disk diffusion method [modified Kirby-Bauer method] on Mueller Hinton Agar (Hi-Media Laboratories, India) following standard zone size interpretative criteria recommended by Clinical and Laboratory Standards Institute (CLSI) [16 ]. Antibiotics that were tested in our study include amoxicillin (AMX 10 μg), amoxicillin clavulanate (AMC 20/10 μg), Gentamycin (GEN 10 μg), Ciprofloxacin (CIP 5 μg), Cotrimoxazole (COT 30 μg), Cefixime (CFM 5 μg), Cefotaxime/Ceftriaxone (CTX/CTR 30 μg), Ceftazidime (CAZ 30 μg), Cefepime (CPM 30 μg), piperacillin tazobactam (PIT 100/10 μg), Imipenem (IMP 10 μg), Meropenem (MEM 10 μg), Polymixin B (PB300 units), and Nitrofurantoin (NI 300 U) (Hi-Media Laboratories, India).
9
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.
10
Antimicrobial Susceptibility Testing of E. coli
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“Antimicrobial susceptibility testing” (AST) was performed using modified Kirby–Bauer disk diffusion method following the guidelines of Clinical and Laboratory Standards Institute guidelines (CLSI).33 Sixteen common antibiotic discs from HiMedia: meropenem (10µg), amoxycillin (10µg), cefotaxime (30µg), cefixime (30µg), ceftriaxone (30µg), ceftazidime (30µg), cefoxitin (30µg), cotrimoxazole (25µg), nitrofurantoin (300µg), levofloxacin (5µg), amikacin (30µg), gentamicin (10µg), cefoperazone/sulbactam (75/30µg), piperacillin/tazobactam (100/10µg), imipenem (10µg), doxycycline (30µg) were used. E. coli that were resistant to at least one agent in three or more antibiotic categories were characterized as multidrug-resistant E. coli.34 (link)
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