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.
Fosfomycin
Manufactured by HiMedia
Sourced in India
Fosfomycin is a laboratory reagent used for research purposes. It is an antibiotic that inhibits bacterial cell wall synthesis. The core function of Fosfomycin is to serve as a tool for studying bacterial growth and susceptibility to this antimicrobial agent.
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Lab products found in correlation
Ciprofloxacin, by HiMedia (3 mentions)
Nitrofurantoin, by HiMedia (3 mentions)
Ampicillin, by HiMedia (3 mentions)
Amikacin, by HiMedia (3 mentions)
Minocycline, by HiMedia (2 mentions)
Vancomycin, by HiMedia (2 mentions)
Tobramycin, by HiMedia (2 mentions)
Cefuroxime, by HiMedia (2 mentions)
Gentamicin, by HiMedia (2 mentions)
Ceftazidime, by HiMedia (2 mentions)
Imipenem, by HiMedia (2 mentions)
Meropenem, by HiMedia (2 mentions)
Piperacillin tazobactam, by HiMedia (2 mentions)
Teicoplanin, by HiMedia (1 mentions)
Clarithromycin, by HiMedia (1 mentions)
Methicillin, by HiMedia (1 mentions)
Streptomycin, by HiMedia (1 mentions)
Tetracycline, by HiMedia (1 mentions)
Kanamycin, by HiMedia (1 mentions)
Norfloxacin, by HiMedia (1 mentions)
5 protocols using fosfomycin
1
Antibiotic Susceptibility Profiling of A. hydrophila
2
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 ].
3
Phenotypic Detection of ESBL and Carbapenem Resistance
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ESBL production was confirmed phenotypically, using guideline M31-A3 of the Clinical and Laboratory Standards Institute (CLSI) (20 ). Resistance to carbapenems was determined by using Etest (HiMedia), and susceptibility was defined by breakpoints per CLSI (20 ). All E. coli isolates were tested for resistance to six major classes of non-β-lactam antibiotics, employing standard disc agar diffusion on Muller-Hinton agar (HiMedia) (20 ). Six antimicrobial discs (HiMedia) were used: ciprofloxacin (30 µg), chloramphenicol (30 µg), fosfomycin (200 µg), gentamicin (10 µg), sulfamethoxazole-trimethoprim (25 µg), and tetracycline (30 µg). Isolates showing resistance to three or more antimicrobial drugs are considered multidrug resistant (MDR).
4
Antibiotic Susceptibility Testing of Bacterial Isolates
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The susceptibility of bacterial isolates against recommended antibiotics was tested by the modified Kirby–Bauer disk diffusion method on Mueller Hinton agar (HiMedia, India) following guidelines of Clinical and Laboratory Standards Institute (CLSI), Wayne, USA [9 ]. Antibiotics that were tested in our study include amoxycillin clavulanate (amc 20/10 μg), ampicillin (amp 10 μg), amikacin (ak 30 μg), ceftazidime (caz 30 μg), ceftazolin (30 μg), cefuroxime (cfm 30 μg), ciprofloxacin (cip 5 μg), cloxacillin (cox 30 μg), cotrimoxazole (cot 25 μg), fosfomycin (fo 200 μg), gentamicin (gen 10 μg), imipenem (imp 10 μg), meropenem (mrp 10 μg), nitrofurantion (300 μg), piperacillin-tazobactam (pit 100/10 μg), teteracycline (te 30 μg), and vancomycin (VAN 30 μg) (HiMedia Laboratories, India). Further, elucidations of antibiotic susceptibility results were made conferring to the zone size interpretative standards of CLSI [9 ]. MDR isolates, resistant to at least one antimicrobial from three different groups of first-line drugs, tested were identified according to the combined guidelines of the European Centre for Disease Prevention and Control (ECDC) and the Centers for Disease Control and Prevention (CDC) [10 (link)]. Escherichia coli ATCC 25922, Staphylococcus aureus (ATCC 25923), and Pseusdomonas aeruginosa (ATCC 27853) were used as a control organism for antibiotic susceptibility testing.
5
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.
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