Amikacin

Manufactured by Thermo Fisher Scientific

Sourced in United Kingdom, United States, Ireland

Amikacin is an antibiotic medication used to treat bacterial infections. It is a type of aminoglycoside antibiotic that works by inhibiting protein synthesis in bacteria, leading to their death or inhibition of growth. Amikacin is commonly used to treat infections caused by Gram-negative bacteria, such as Pseudomonas, Acinetobacter, and Klebsiella.

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154 protocols using amikacin

Antimicrobial Susceptibility Testing of Pathogens

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To test antibiotic resistance in Campylobacter spp., the broth microdilution method was used with 5 % sheep blood. For all other pathogens, antimicrobial susceptibilities were determined by the agar dilution method according to the Clinical and Laboratory Standards Institute (CLSI) Guidelines, 2015 [20 ]. All isolates of Salmonella spp. were tested for their minimum inhibitory concentrations (MICs) of ampicillin, ampicillin-sulbactam, ceftriaxone, cefotaxime, nalidixic acid, ciprofloxacin, levofloxacin, co-trimoxazole, azithromycin, chloramphenicol and tetracycline (Oxoid); DEC were tested for ampicillin, ampicillin-sulbactam, cefotaxime, ciprofloxacin, levofloxacin, chloramphenicol, tetracycline, cefazolin, cefuroxime, imipenem, amikacin and gentamicin (Oxoid); Campylobacter spp. were tested for ciprofloxacin, azithromycin, tetracycline, erythromycin and doxycycline (Oxoid); and Aeromonas spp. were tested for cefotaxime, ciprofloxacin, levofloxacin, co-trimoxazole, chloramphenicol, tetracycline, cefazolin, cefuroxime, imipenem, amikacin and gentamicin (Oxoid). ATCC 25922, 35218, 700603 and 27853 were used as quality control strains. Antibiotic susceptibility was interpreted according to CLSI guidelines, 2015 [20 ].

Tian L., Zhu X., Chen Z., Liu W., Li S., Yu W., Zhang W., Xiang X, & Sun Z. (2016). Characteristics of bacterial pathogens associated with acute diarrhea in children under 5 years of age: a hospital-based cross-sectional study. BMC Infectious Diseases, 16, 253.

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Antimicrobial Susceptibility Testing Protocol

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Antimicrobial susceptibility was determined using the disc diffusion method, in accordance with the Clinical and Laboratory Standard Institute (CLSI) [27 (link)] for the following antibiotics: Ampicillin (AMP, 10 µg), cefoxitin (FOX, 30 µg), cefotaxime (CTX, 30 µg), ciprofloxacin (CIP, 5 µg), norfloxacin (NOR, 10 µg), tobramycin (TOB, 10 µg), gentamicin (GEN, 10 µg), doxycycline (DOX, 5 µg), amikacin (AMK, 30 µg), azithromycin (AZM, 15 µg), nitrofurantoin (NIT, 300 µg), amikacin, vancomycin (VA, 30 µg), linezolid (LZD, 30 µg) and sulfamethoxazole/trimethoprim (SXT, 1.25/23.75 µg) (Oxoid, Hampshire, England). The results were interpreted using the criteria outlined in CLSI guidelines based on the inhibition zone produced, which correlate with susceptibility levels [27 (link)]. The obtained results were used to identify the percentage of MDR among the tested isolates. As previously documented, multidrug resistance (MDR) is defined as resistance to three or more antimicrobial classes [28 (link)]. The phenotypic identification of the isolates as MRSA was performed against cefoxitin through the disk diffusion method, while the standard strain of S. aureus (ATCC 29312) was included as a control isolate.

Ramadan H.A., El-Baz A.M., Goda R.M., El-Sokkary M.M, & El-Morsi R.M. (2023). Molecular characterization of enterotoxin genes in methicillin-resistant S. aureus isolated from food poisoning outbreaks in Egypt. Journal of Health, Population, and Nutrition, 42, 86.

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Hydrated Antibiotic-Loaded Bone Cement

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Combinations of TMC and PEGDAc and non-derivatized paste were hydrated using a 5 mg/mL combination solution of vancomycin and amikacin (MP Biomedicals, USA). Approximately 0.6 mL of hydrated paste (n = 4) was injected into cell crowns (Scaffdex, Finland) with nylon filters (pore size = 41 μm) attached (See Supplementary Material for visual representation). Each sample was placed in 5 mL of phosphate buffered saline (PBS), incubated at 37°C, and sampled daily. Upon sampling, each sample was completely refreshed with PBS. vancomycin was detected and quantified using a high performance liquid chromatography (Dionex UltiMate 3000 HPLC, Thermo Scientific, Waltham, MA) system interfaced with a UV/Vis spectrophotometer at 209 nm [32 (link)]. amikacin was quantified using a previously described method of pre-column derivatization with an o-phthaldialdehyde reagent (AdipoGen Life Sciences, USA) and subsequent detection with an HPLC system using a fluorescence detector (Excitation = 340 nm, Emission = 455 nm) [33 ]. Both detections utilized reverse-phase columns, C18 150 × 4.6 mm (Hypersil Gold, Thermo Scientific) for vancomycin and C8 100 × 4.6 mm (Hypersil BDS, Thermo Scientific) for amikacin, with mobile phase consisting of 85% phosphate buffer at pH = 7.4 and 15% acetonitrile.

Boles L.R., Bumgardner J.D., Fujiwara T., Haggard W.O., Guerra F.D, & Jennings J.A. (2019). Characterization of trimethyl chitosan/polyethylene glycol derivatized chitosan blend as an injectable and degradable antimicrobial delivery system. International journal of biological macromolecules, 133, 372-381.

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Antibiotic Susceptibility Testing of Staphylococcus aureus

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The following antibiotics (Thermo Fisher Scientific Inc., Washington, USA) were used for antibiotic susceptibility testing of S. aureus:
Amikacin (AK), cefoxitin (FOX), chloramphenicol (C), co-trimoxazole (SXT), ciprofloxacin (CIP), gentamicin (CN), linezolid (LZD), fusidic acid (FD), neomycin, norfloxacin (NOR), tetracycline (TE), penicillin (P), clindamycin (DA), erythromycin (E), tigecycline (TGC), teicoplanin (TEC), tobramycin (TOB), and vancomycin (VA).
All of the coagulase test positive staphylococci isolates were tested for resistance to FOX on MH Agar using 30 ug/mL FOX according to the standard guidelines of CLSI. As per CLSI guidelines, the resistant strains showed a zone diameter of <24 mm, while a diameter of >25 mm showed the susceptible (S) strains. Strains that were resistant to FOX were noted as MRSA [24 ].

Ahmed N., Khalid H., Mushtaq M., Basha S., Rabaan A.A., Garout M., Halwani M.A., Al Mutair A., Alhumaid S., Al Alawi Z, & Yean C.Y. (2022). The Molecular Characterization of Virulence Determinants and Antibiotic Resistance Patterns in Human Bacterial Uropathogens. Antibiotics, 11(4), 516.

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Colistin Resistance Determination in E. coli

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Antibiotic susceptibility was determined by disk diffusion method. The tested antibiotics were cefoxitin, ceftriaxone, ertapenem, imipenem, meropenem, gentamicin, amikacin, nalidixic acid, ciprofloxacin, tetracycline, erythromycin, piperacillin/tazobactam, cefoperazone/sulbactam and colistin (Thermo Fisher Scientific, USA). colistin resistance was further confirmed by the broth microdilution method. The minimum inhibitory concentrations (MICs) of colistin were determined by the broth microdilution in cation-adjusted Mueller–Hinton II broth according to Clinical and Laboratory Standards Institute (CLSI) 2017 guidelines [16 ]. E. coli ATCC 25922 was used as a control and a range of colistin dilutions (Chem-Impex Int’l Inc., USA) between 0.25 mg/L and 128 mg/L were performed. Breakpoints of colistin susceptibility defined by European Committee on Antimicrobial Susceptibility Testing (EUCAST) [17 ] were used as follows: isolates with a colistin MIC ≤ 2 mg/L were categorised as susceptible, and those with a colistin MIC > 2 mg/L were categorised as resistant.

Boonyasiri A., Brinkac L.M., Jauneikaite E., White R.C., Greco C., Seenama C., Tangkoskul T., Nguyen K., Fouts D.E, & Thamlikitkul V. (2023). Characteristics and genomic epidemiology of colistin-resistant Enterobacterales from farmers, swine, and hospitalized patients in Thailand, 2014–2017. BMC Infectious Diseases, 23, 556.

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Antibiotic Susceptibility Testing Protocol

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The antibacterial susceptibility testing was performed usingthe disk diffusion and the VITEK 2 system (bioM’erieux, Marcy l’Etoile, France) and interpreted by the protocol specified in the guidelines of Clinical and Laboratory Standards Institute (CLSI, 2015). The antibacterial agents included cefoxitin, ceftriaxone, cefotaxime, cefepime, ceftazidime, cefazolin, moxifloxacin, ofloxacin, levofloxacin, gentamicin, amikacin, amoxycillin, minocycline, piperacillin, azithromycin, nitrofurantoin, polymyxin B, and meropenem (Thermo Fisher Scientific, USA). MDR isolates were defined as resistant to at least three different classes of antibiotics. Isolates were defined as extensively drug resistant (XDR) if they were not susceptible to at least one agent in all but two or fewer antibacterial categories (ie, bacterial isolates remained susceptible to only one or two antibacterial categories).

Qian W., Li X., Yang M., Liu C., Kong Y., Li Y., Wang T, & Zhang Q. (2022). Relationship Between Antibiotic Resistance, Biofilm Formation, and Biofilm-Specific Resistance in Escherichia coli Isolates from Ningbo, China. Infection and Drug Resistance, 15, 2865-2878.

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Antibiotic Susceptibility of Lactobacillus from Bees

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The antibiotic susceptibility of Lactobacillus strains isolated from the GI tract of bees was evaluated using the following antibiotics: Cefotaxime, Ampicillin, Gentamicin, Chloramphenicol, Kanamycin, Amikacin,-sSulfamethoxazole-Trimethoprim, Ciprofloxacin, Colistin, Rifampicin, Vancomycin, Neomycin, Streptomycin, Efrotomycin, Clarithromycin and Penicillin (Oxoid, Thermo Fisher Scientific, Waltham, MA, USA). Bacterial suspensions of the strains to be assayed were prepared at a concentration equivalent to 0.5 McFarland in sterile saline solution. One hundred μL of the bacterial suspension were seeded on MRS agar containing dishes and a maximum of 5 antibiotic disks were placed per dish. Then, dishes were incubated under microaerobic conditions at 37 °C for 48 h. After 48 h incubation, the zones of inhibition were measured and recorded in mm. The criterium to evaluate susceptibility was the one described by (Georgieva et al., 2008 (link)) expressing sensitivity as R (resistant); MS (intermediate sensitive)- zone of inhibition between 7 and 16 mm; S (sensitive) zone of inhibition between 16 and 25 mm; SS (highly sensitive) zone of inhibition over 25 mm; ND not determined.

Carvajal R.I., Silva-Mieres F., Ilabaca A., Rocha J., Arellano-Arriagada L., Zuniga Arbalti F.A, & García-Cancino A. (2023). Isolation and characterization of Lactobacillus casei A14.2, a strain with immunomodulating activity on Apis mellifera. Saudi Journal of Biological Sciences, 30(4), 103612.

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Preparation and Application of Aminoglycoside Antibiotics

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AGs (Sigma – Gentamicin sulfate, Paromomycin sulfate, Neomycin sulfate, Tobramycin, Amikacin sulfate; Thermo Fisher – G418 sulfate) were prepared by dissolving compounds in either water or 150 mM Tris pH 8.0 to prepare 50 mg/mL stock solutions of each compound. Liquid preparations were also purchased for G418 and Gentamicin (Thermo Fisher Scientific). Activity of compounds was not influenced by preparation methods (data not shown), so compounds prepared in 150 mM Tris pH 8.0 were used for the majority of experiments. Prior to addition of AGs for ribosome profiling and RNA-seq, AGs were added to media and equilibrated in a cell culture incubator to allow stabilization of pH and temperature before addition to cells.

Wangen J.R, & Green R. (2020). Stop codon context influences genome-wide stimulation of termination codon readthrough by aminoglycosides. eLife, 9, e52611.

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Antibiotic Susceptibility of P. aeruginosa

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The following antibiotics (Thermo Fisher Scientific Inc., Waltham, MA, USA) were used for antibiotic susceptibility testing of P. aeruginosa:
Amikacin (AK), ciprofloxacin (CIP), co-trimoxazole (SXT), gentamicin (CN), tobramycin (TOB), aztreonam (AZM), cefepime (FEP), ceftazidime (CAZ), colistin (CT), meropenem (MEM), imipenem (IPM), piperacillin-tazobactam (TZP) and polymyxin (PB) [24 ].

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Synergism Testing of Antibiotics

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The MIC was initially determined using strips impregnated with colistin (bioMérieux, France), amikacin, meropenem (Thermo Fisher Scientific, Basingstoke, UK), and ertapenem (Liofilchem, Italy) at increasing concentrations. For the synergism testing, one test strip was placed on the inoculated MHA plate. After 1 h at room temperature, the agar was marked adjacent to the previously determined MIC of the agent, and the tape was replaced. The second strip was then placed over the imprint of the previous strip such that the mark on the agar corresponds to the MIC of the second agent [20 (link)]. The highest value was considered for isolates in which the MIC exceeded the value of the strip concentration [6 (link)]. The resulting ellipse of inhibition was checked after 18–20 h at 35 ± 2 °C and the Fractional Inhibitory Concentration Index (ΣFIC) was calculated and interpreted using the following criteria: synergism, ΣFIC ≤0.5; antagonism, ΣFIC> 4; and indifferent, ΣFIC> 0.5–4 [6 (link), 15 (link), 20 (link), 31 (link)–40 ].

Gaudereto J.J., Neto L.V., Leite G.C., Espinoza E.P., Martins R.C., Villas Boa Prado G., Rossi F., Guimarães T., Levin A.S, & Costa S.F. (2020). Comparison of methods for the detection of in vitro synergy in multidrug-resistant gram-negative bacteria. BMC Microbiology, 20, 97.

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