Antimicrobial sensitivity testing was done by Kirby-Bauer Antimicrobial Susceptibility Test (disc diffusion method) using Mueller-Hinton agar [15 ]. Six different antibiotics were tested and the zone size was measured. Amikacin, streptomycin, gentamicin, tobramycin, netilmicin, and neomycin (obtained from HiMedia, India) were taken and screening test for detection of high level aminoglycoside resistance (HLAR) in Enterococcus species was confirmed as per Clinical and Laboratory Standards Institute (CLSI) approved standards [16 ] The E. coli isolates were described as isolates with high level aminoglycoside resistance considering growth ≥2048 μg/mL for streptomycin, ≥512 μg/mL for gentamicin, ≥512 μg/mL for neomycin, ≥256 μg/mL for tobramycin, ≥256 μg/mL for netilmicin, and ≥256 μg/mL for Amikacin. MICs were determined by Etest (AB Biodisk).
Neomycin
Manufactured by HiMedia
Sourced in India
Neomycin is a broad-spectrum antibiotic used in laboratory settings. It inhibits bacterial protein synthesis, thereby preventing bacterial growth.
Automatically generated - may contain errors
Lab products found in correlation
Streptomycin, by HiMedia (5 mentions)
Amikacin, by HiMedia (4 mentions)
Ampicillin, by HiMedia (4 mentions)
Tetracycline, by HiMedia (4 mentions)
Gentamicin, by HiMedia (3 mentions)
Chloramphenicol, by HiMedia (3 mentions)
Penicillin g, by HiMedia (3 mentions)
Ceftriaxone, by HiMedia (3 mentions)
Polymyxin b, by HiMedia (2 mentions)
Ciprofloxacin, by HiMedia (2 mentions)
Cefotaxime, by HiMedia (2 mentions)
Amoxicillin, by HiMedia (2 mentions)
Enrofloxacin, by HiMedia (2 mentions)
Etest, by bioMérieux (1 mentions)
Netilmicin, by HiMedia (1 mentions)
Tobramycin, by HiMedia (1 mentions)
Cephalexin, by HiMedia (1 mentions)
Cotrimoxazole, by HiMedia (1 mentions)
Norfloxacin, by HiMedia (1 mentions)
Gentamycin, by HiMedia (1 mentions)
7 protocols using neomycin
1
Antimicrobial Resistance Profiling in Enterococcus
2
Antibiotic Susceptibility Profiling of Vibrio
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Antibiotic susceptibility testing was implemented in accordance with the Kirby–Bauer disk diffusion method following the Clinical and Laboratory Standards Institute (CLSI) guidelines [47 (link)]. In brief, 150 µL of overnight Vibrio culture was spread aseptically on LB agar plates. E. coli DH5α was used as the negative control (usually E. coli strain K12 MG1655 is used as a negative control). The antibiotics which are most commonly used for the treatment of cholera were chosen for antibiotic susceptibility testing [15 (link),33 (link)]. Different antibiotics discs [ampicillin (AMP, 10 μg), chloramphenicol (C, 30 μg), cefotaxime (Ce, 30 μg), ciprofloxacin (Cf, 5 μg), co-trimoxazole (Co, 25 μg), polymyxin-B (PB, 300 unit), furazolidone (FR, 100 μg), gentamycin (GEN, 10 μg), neomycin (N, 30 μg), norfloxacin (Nx, 10 μg), nalidixic acid (Na, 30 μg), tetracycline (T, 30 μg), cephalexin (CN, 30 μg), streptomycin (S, 10 μg), trimethoprim (Tr, 5 μg)] (HiMedia, Mumbai, India) were placed at certain distances on the surface of the agar plate. The plates were incubated at 37 °C for 24 h and then the diameter of the zone of inhibition of each antibiotic disc was measured.
3
Antibiotic Susceptibility Testing Protocol
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The antibiotic susceptibility test was conducted using the Kirby–Bauer disk diffusion method [37 (link)]. The standard antibiotic discs were procured from ‘HiMedia, Mumbai, India, which include polymyxin-B (300 µg), amoxiclav (30 µg), rifampicin (5 µg), tetracycline (30 µg), oxacillin (5 µg), amikacin (30 µg), cefoxitin (30 µg), cefepime (30 µg), ceftazidime (30 µg), cefotaxime (30 µg), chloramphenicol (30 µg), cefdinir (5 µg), penicillin g (10 µg), moxifloxacin (5 µg), ampicillin (10 µg), vancomycin (30 µg), ceftriaxone (30 µg), neomycin (10 µg), ofloxacin (5 µg), norfloxacin (10 µg), kanamycin (30 µg), bacitracin (10 µg), co-trimoxazole (25 µg), methicillin (10 µg), streptomycin (10 µg), levofloxacin (5 µg), erythromycin (15 µg), clindamycin (2 µg), gentamycin (120 µg), and sterile disc (control). The antibiotic discs were placed onto the freshly prepared lawns of each isolate on Mueller–Hinton agar (MHA) plates. The plates were incubated at 40 °C for 24–48 h, and the diameter of the zone of inhibition was measured in millimetres. The strains were classified in accordance with the Clinical and Laboratory Standards Institute [38 ], following the standard antibiotic disc chart.
4
Antibiotic Sensitivity of Listeria Isolates
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All Listeria isolates were subjected to antibiotic sensitivity test against 12 different antimicrobial agents by agar diffusion method in Mueller-Hinton AGAR [15 ]. Listeria isolates were tested against amoxicillin (10 mg), cefixime (10 mg), chloramphenicol (25 mg), furazolidone (100 mg), co-trimoxazole (25 mg), oxytetracycline (30 mg), erythromycin (15 mg), chlortetracycline (30 mg), streptomycin (10 mg), and nitrofurazone (100 mg), neomycin (10 mg), doxycycline hydrochloride (30 mg), and antibiotic discs (HiMedia, Mumbai). The clinical breakpoints for Listeria susceptibility testing were defined according to the Clinical and Laboratory Standard Institute [16 ] and the isolates were grouped as sensitive, intermediary sensitive, and resistant against each antibiotic.
5
Antimicrobial Resistance Profiling of Urinary Isolates
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Different strains of various organisms isolated from urine samples of the infected animals were subjected to in vitro drug sensitivity testing, using 15 antimicrobials by the disc-diffusion method. With the help of a platinum loop, a small amount of test culture was transferred into a tube of brain heart infusion broth and incubated for 2-5 h at 37°C, to obtain turbidity. With the help of a sterile cotton swab, the broth culture was then evenly spread by smearing over the surface of BA/Mueller-Hinton agar plates. The antimicrobial discs were placed on the agar and gently pressed. These were then incubated at 37°C for 24 h. The sensitivity was observed on the basis of zone size interpretation chart, provided by the manufacturer. Different antimicrobials used were amikacin (10 mcg), amoxicillin (10 mcg), amoxicillin-clavulanic acid (30 mcg), ampicillin (25 mcg), cefoperazone (75 mcg), ceftriaxone (10 mcg), ceftriaxone–tazobactam (30mcg), chloramphenicol (25 mcg), cloxacillin (30 mcg), enrofloxacin (10 mcg), gentamicin (30 mcg), neomycin (30 mcg), oxytetracycline (30 mcg), penicillin-G (10 units), and streptomycin (25 mcg) (Hi-Media). To remain conservative in our estimates of resistance, isolates exhibiting intermediate zones of inhibition were interpreted as resistant.
6
Antibiotic Susceptibility of SMB1 and DSM 19409
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The susceptibility of strain SMB1T and the DSM 19409T to different antibiotics was checked using disc diffusion assay as per Clinical and Laboratory Standards Institute guidelines41 . The antibiotics discs (Hi-Media, India) used were Tetracycline (30 mcg), Neomycin (30), Penicillin G (2 units), Gentamycin (10 mcg), Kanamycin (30 mcg), Amoxycillin (30 mcg), Cephanroxcil (30 mcg), Lincomycin (2 mcg), Cefalexin (30 mcg), Chloramphenicol (30 mcg), Cefazolin (30 mcg), Polymixin B (300 units), Cefprozil (30), Vancomycin (30 mcg), Novobiocin (30 mcg) and ChlorTetracycline (30 mcg).
7
Antibiotic Sensitivity Patterns of Isolates
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In vitro antibiotic sensitivity patterns of the isolates were conducted as per the method of Bauer et al., (1966) . Antibiotics disc (Hi Media Ltd., Mumbai, India) used in the present study were Amikacin (30 mcg), Amoxyclav (30 mcg), Ampicillin (10 mcg), Ciprofloxacin (5 mcg), Colistin (10 mcg), Ceftriaxone (30 mcg), Erythromycin (15 mcg), Enrofloxacin (10 mcg), Gentamicin (10mcg), Neomycin (30 mcg), Penicillin-G (10IU), Streptomycin (10 mcg), Sulphadiazine (300 mcg) and Tetracycline (30 mcg). Diameters of the clear zone of inhibition were measured and the interpretation of the results was made in accordance with the instructions supplied by the manufacturer (Hi Media Ltd., Mumbai, India). Multiple Antibiotic resistance index (MARI) were also determined for each isolates by dividing the number of antibiotics to which the isolate is resistant to by the total numbers of antibiotics tested (Adenaike, 2016) .
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