Resistance to antibiotics was confirmed as per the guidelines of internationally recognized standards of the Clinical and Laboratory Standards Institute (CLSI)53 . MRSA, MRSE and MRSH strains were screened for susceptibility by 22 antimicrobial agents using disc diffusion method on Muller-Hinton agar. The antibiotics disc tested were (concentration in μg, HiMedia, India) Erythromycin (E-15 μg), Clindamycin (CD-2 μg), Gentamicin (GEN-10 μg), Cefotaxime (CTX-30 μg), Norfloxacin (NX-10 μg), Ciprofloxacin (CIP-5 μg), Tetracycline (TE-30 μg), Rifampicin (RIF-5 μg), Nitrofurantoin (NIT-300 μg), Linezolid (LZ-30 μg), Chloramphenicol (C-30 μg), Amoxyclav (AMC-10 μg), Amphicillin (AMP-10 μg), Azithromycin (AZM-15 μg), Bacitracin (B-10 Units), Cefoxitime (CFM-5 μg), co-Trimoxazole (COT-23.75 μg), Kanamycin (K- 30 μg), Oxacillin (OX-I μg), Penicillin G (P-10 Units), Streptomycin (S-100 μg), Tetracycline (TE-30 μg) and Tobramycin (TOB-30 μg). The results were recorded after 24 h of incubation at 37 °C and interpreted as per CLSI guidelines. Screened isolates showing resistance to more than three classes of non-β-lactam antibiotics were considered as multidrug resistant (MDR) strains, as described by Chajecka et al.,9 (link). Assay was performed in triplicates for all strains.
Tetracycline
Manufactured by HiMedia
Sourced in India
Tetracycline is a broad-spectrum antibiotic used in laboratory settings. It functions as a protein synthesis inhibitor, preventing bacterial cells from producing proteins essential for their growth and survival.
Automatically generated - may contain errors
Lab products found in correlation
Ciprofloxacin, by HiMedia (43 mentions)
Gentamicin, by HiMedia (33 mentions)
Ampicillin, by HiMedia (32 mentions)
Chloramphenicol, by HiMedia (31 mentions)
Erythromycin, by HiMedia (26 mentions)
Cefotaxime, by HiMedia (22 mentions)
Cotrimoxazole, by HiMedia (21 mentions)
Ceftriaxone, by HiMedia (19 mentions)
Streptomycin, by HiMedia (19 mentions)
Amikacin, by HiMedia (17 mentions)
Nalidixic acid, by HiMedia (17 mentions)
Ceftazidime, by HiMedia (15 mentions)
Vancomycin, by HiMedia (14 mentions)
Norfloxacin, by HiMedia (14 mentions)
Cefoxitin, by HiMedia (14 mentions)
Imipenem, by HiMedia (13 mentions)
Amoxicillin, by HiMedia (13 mentions)
Levofloxacin, by HiMedia (12 mentions)
Gentamycin, by HiMedia (12 mentions)
Clindamycin, by HiMedia (12 mentions)
71 protocols using tetracycline
1
Antimicrobial Resistance Profiling of MRSA
2
Antibiotic Susceptibility Testing of Shigella
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Antibiotic susceptibility testing was done using Kirby-Bauer disc diffusion method (3 ) for the antibiotics: ampicillin 10 µg, trimethoprim-sulphamethoxazole 1.25/23.75 µg, ciprofloxacin 5 µg, ceftriaxone 30 µg, tetracycline 30 µg, and chloramphenicol 30 µg (Himedia Laboratories, Mumbai). The minimum inhibitory concentration (MIC) for ciprofloxacin and ceftriaxone were performed using Epsilometer test (E-test) strips according to the manufacturer's instructions (AB Biomeriuex, India). The inoculum for the susceptibility testing and the interpretation were done as per CLSI (Clinical Laboratory Standards Institute) guidelines (3 ). ATCC Escherichia coli 25922 was used as the control for interpretation of zone diameters. Combination disc method according to CLSI guidelines was used in order to detect ESBL production in ceftriaxone-resistant Shigella isolates.
3
Antibiotic Susceptibility Testing of Yeast Isolates
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Isolated yeast strains were tested against 30 antibiotics with different modes of actions such as amikacin (MD001), amoxycillin (MD002), azithromycin (MD004), benzyl penicillin (MD062), cefalexin (cephalexin) (MD014), cefepime (MD070), cefotaxime (cephotaxime) (MD064), chloramphenicol (MD016), ciprofloxacin (MD017), erythromycin (MD022), gemifloxacin (MD076), gentamicin (MD061), kanamycin (MD026), levofloxacin (MD027), methicillin (MD031), moxifloxacin (MD033), neomycin (MD036), norfloxacin (MD038), ofloxacin (MD039), pefloxacin (MD040), polymyxin-B (MD043), rifampicin (MD045), roxithromycin (MD046), streptomycin (MD048), sulphadiazine (MD050), sulphamethizole (MD052), teicoplanin (MD055), vancomycin (MD060), tetracycline (MD056), meropenem (SD727) (Himedia, Mumbai, India) with standard antibiotic concentration previously determined by testing against pathogens [17 (link)]. The diameter of the inhibition zone was measured after 48 h of incubation at 30 °C.
4
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) .
5
Antibiotic Susceptibility Screening of Isolates
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Antibiotic drug susceptibility was determined by spreading overnight grown culture of the isolates on MRS agar plates as a lawn. Standard antibiotic discs (tetracycline, erythromycin, ampicillin, gentamycin, penicillin, chloramphenicol, cefuroxime, cefoperazone, levofloxacin, norfloxacin, Hi-Media, Mumbai) were placed on the surface of the MRS agar medium aseptically. Plates were incubated for 24 h at 37 °C and observed for zones of inhibition.
6
Antibiotic Susceptibility of E. coli
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All E. coli isolates were tested for antibiotics susceptibility by using the disk diffusion method as reported by Kirby-Bauer [18 (link)]. Briefly, isolates were suspended in sterile 0.85% normal saline and adjusted to 0.5 McFarland standard solution. Then, MHA plates were inoculated, and antibiotic disks were seeded within 15 min after inoculation of MHA plates. MHA plates were incubated aerobically at 37 °C for 16–18 h. The interpretations of zones of inhibitions were performed as recommended by the CLSI 29th Edition guidelines [19 ]. All E. coli that showed intermediate susceptibility to the antibiotics tested were regarded as resistant to such particular antibiotics. Antibiotics tested included ciprofloxacin (CIP 5 μg; HiMedia, Mumbai, India), ampicillin (AMP 10 μg; HiMedia, India), tetracycline (TE 30 μg; HiMedia, India), meropenem (MEM 10 μg; HiMedia, India), ceftazidime (CAZ 30 μg; HiMedia, India), gentamicin (CN 10 μg; HiMedia, India), cefepime (FEP 30 μg; HiMedia, India), and trimethoprim-sulfamethoxazole (SXT 25 μg; HiMedia, India).
7
Antibiotic Susceptibility of Gram-Negative Bacteria
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The susceptibilities of Gram-negative bacteria (GNB) to the antimicrobial agents ampicillin (10 µg) amoxicillin/clavulanate (30 µg), cefuroxime (30 µg), cefotaxime (30 µg), chloramphenicol (30 µg), cotrimoxazole (25 µg), gentamicin (10 µg), amikacin (30 µg), ciprofloxacin (5 µg), tetracycline (30 µg) and levofloxacin (5 µg) (HiMedia Laboratories, India) were determined with Kirby-Bauer disc diffusion method in accordance with the Clinical and Laboratory Standards Institute (CLSI) guidelines [16 ]. Erythromycin (30 µg), oxacillin (30 µg), and vancomycin (30 µg) were included for Gram positive bacteria (GPB). The reference strain E. coli ATCC 25922 and Staphylococcus aureus ATCC 10221 were included as quality controls in the susceptibility assays. Relative to the panel of antibiotics tested for each isolate, and according to the international standard definitions for acquired resistance, multidrug resistant (MDR) phenotype was defined as in vitro non-susceptibility to ≥1 agent in ≥3 antimicrobial categories [17 (link)]: penicillins, cephalosporins, beta-lactamase inhibitor combinations, fluoroquinolones, aminoglycosides, chloramphenicol, folate pathway inhibitors, tetracyclines, macrolides and glycopeptides.
8
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.
9
Antibiotic Susceptibility of E. coli and Salmonella
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Antibiotic sensitivity testing of isolated E. coli and Salmonella spp. was carried out using the disk diffusion assay as previously described [63 (link)]. Antibiotic classes included fluoroquinolones (levofloxacin, LEV—5 μg; ciprofloxacin, CIP—5 μg), aminoglycosides (gentamicin, GEN—10 μg; streptomycin, S—10 μg), carbapenems (Meropenem, MEM—10 μg; imipenem, IMP—10 μg), amphenicols (chloramphenicol, C—10 μg), macrolides (erythromycin, E—15 μg), and tetracyclines (tetracycline, TE—30 μg) purchased from Hi Media (India). Sensitivity tests were performed on freshly grown isolates having a concentration equivalent to 0.5 McFarland standard using Mueller-Hinton agar media (Hi Media, India). All results were interpreted according to the guidelines provided by Clinical and Laboratory Standards Institute [64 ]. Furthermore, isolates showing resistance against three or more different classes of antibiotics were defined as MDR [65 (link)].
10
Antibiotic Susceptibility Testing for Klebsiella
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Antibacterial susceptibility testing was performed using the disk diffusion method according to the guidelines of the Clinical and Laboratory Standards Institute (CLSI) documents (13 ). The antibiotics used comprised of piperacillin (100 μg), ceftazidime (30 μg), cefotaxime (30 μg), cefazoline (30 μg), tetracycline (30 μg), kanamycin (30 μg), Imipenem (10 μg), and Meropenem (10 μg) (Himedia, Mombay, India). Minimum inhibitory concentration (MIC) for Imipenem was determined by E-test method (AB Biodisk, Solna, Sweden) for all K. pneumoniae isolates according to CLSI guideline (14 (link)).
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