Antimicrobial susceptibility was assessed by the disk diffusion method according to Clinical and Laboratory Standards Institute (CLSI) guidelines [8 9 ]. For 34 E. faecalis and 32 E. faecium isolates, 14 antibiotics were used to test susceptibility: ampicillin (AM), 10 µg; amoxicillin-clavulanic acid (AMC), 30 µg; chloramphenicol (C), 30 µg; tetracycline (TE), 30 µg; ciprofloxacin (CIP), 5 µg; erythromycin (E), 15 µg; imipenem (IPM), 10 µg; sulfamethoxazole/trimethoprim (SXT), 23.75 µg/1.25 µg; gentamycin (GM), 10 µg; streptomycin (S), 10 µg; high-level gentamicin (HLGM), 120 µg; high-level streptomycin (HLS), 300 µg; kanamycin (K), 30 µg; and quinupristin/dalfopristin (SYN), 15 µg (BD Biosciences). Each cultured single isolate was suspended in 0.85% NaCl and turbidity was adjusted to 0.5 McFarland. Five antibiotic disks were placed on each Mueller-Hinton agar (BD Biosciences) plate followed by incubation at 37℃ for 24 h. The diameters of the inhibitory zone of the disks were measured and tested isolates were determined to be susceptible, intermediate, or resistant based on the CLSI M100-S24 E. faecalis and E. faecium breakpoint guidelines. Interpretive criteria were adapted from Enterobacteriaceae guidelines as published in CLSI M100-S23 when the breakpoint was unavailable. S. aureus ATCC 25923 and E. faecalis ATCC 29212 were used as controls.
Erythromycin
Manufactured by BD
Sourced in United States, Germany
Erythromycin is a macrolide antibiotic used in various laboratory applications. It is a natural product derived from the bacterium Saccharopolyspora erythraea. Erythromycin inhibits bacterial protein synthesis by binding to the 50S subunit of the bacterial ribosome.
Automatically generated - may contain errors
Lab products found in correlation
Clindamycin, by BD (14 mentions)
Tetracycline, by BD (14 mentions)
Ciprofloxacin, by BD (13 mentions)
Gentamicin, by BD (13 mentions)
Chloramphenicol, by BD (10 mentions)
Cefoxitin, by BD (8 mentions)
Penicillin, by BD (8 mentions)
Trimethoprim sulfamethoxazole, by BD (7 mentions)
Fusidic acid, by BD (6 mentions)
Tobramycin, by BD (6 mentions)
Rifampicin, by BD (6 mentions)
Vancomycin, by BD (6 mentions)
Kanamycin, by BD (5 mentions)
Rifampin, by BD (5 mentions)
Linezolid, by BD (5 mentions)
Streptomycin, by BD (5 mentions)
Mupirocin, by BD (4 mentions)
Imipenem, by BD (4 mentions)
Novobiocin, by BD (4 mentions)
Vancomycin, by bioMérieux (3 mentions)
26 protocols using erythromycin
1
Antimicrobial Susceptibility in Enterococcus
2
Antibiotic Susceptibility Testing of Isolates
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The antibiotic susceptibility of the all isolates was determined by the disk diffusion method on Mueller–Hinton agar (Becton–Dickinson, Sparks, MD, USA) according to the recommendations of the Clinical and Laboratory Standards Institute (CLSI). The disk diffusion assay was run with 15 antibiotics: bacitracin (10 U), enrofloxacin (5 μg) (OXOID), streptomycin (10 μg), spiramycin (100 μg), sulfadiazine (25 μg), chloramphenicol (30 μg) (BD), doxycycline (30 μg), erythromycin (15 μg), gentamicin (10 μg), neomycin (30 μg), penicillin (10 U), tetracycline (30 μg), cefoxitin (30 μg), trimethoprim/sulfamethoxazole (1.25 μg/23.75 μg, respectively), and vancomycin (30 µg and 5 µg) (BIO-RAD, Hercules, CA, USA). S. aureus strain ATCC 25923 and Enterococcus faecalis strain ATCC 29212 were used as controls in the susceptibility test.
3
Identification and Antimicrobial Susceptibility of S. aureus
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S. aureus isolates were identified by MALDI-TOF mass spectrometry (Bruker Daltonic GmBH, Bremen, Germany), and MRSA isolates were detected by antibiotic susceptibility testing using the agar disk diffusion method according to the European Committee on Antimicrobial Susceptibility Testing (EUCAST) [18 ]. The antibiotics tested were penicillin, cefoxitin, gentamicin, erythromycin, clindamycin, tetracycline, ciprofloxacin, trimethoprim-sulfamethoxazole, rifampin, linezolid, and mupirocin (BD, Sparks, USA). The minimal inhibitory concentration (MIC) of vancomycin and teicoplanin was determined using the E-test (bioMerieux, Marcy-l’Etoile, France).
4
Bacterial Strain and Cultivation Protocols
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Bacterial strains used in this study are listed in Table 1 .Table 1 .
5
Antimicrobial Susceptibility Testing of Bacterial Isolates
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An antimicrobial susceptibility test was performed for each pure isolate of bacteria on Mueller Hinton agar by using Kirby-Bauer disk diffusion techniques.27 Accordingly, detailed Clinical and Laboratory Standards Institute (CLSI) guidelines.25 For each category of gram-positive and gram-negative bacteria isolates were tested against, amoxicillin (30 µg, BD), amoxycillin-clavulanic acid (30 µg, BD), ceftazidime (30 µg, BD), cefotaxime (30 µg, BD), ceftriaxone (30 µg, BD), chloramphenicol (30 µg, BD), clindamycin (2 µg, BD), gentamicin (10 µg, BD), cotrimoxazole (25 μg), tetracycline (30 µg, BD), ciprofloxacin (5 µg, BD), penicillin (10 units, BBL), cefoxitin (30 μg), erythromycin (15 µg, BD), piperacillin (100 μg), cefepime (30μg), streptomycin (10 μg) and meropenem (10 μg). Then after overnight incubation at 37°C, the diameter of zone of inhibition was measured by millimetre and interpreted as sensitive, intermediate and resistant.25
6
Bacillus cereus Biofilm Cultivation
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B. cereus was routinely grown at 25°C or 37°C in LB broth or on LB agar (BD, Franklin Lakes, NJ, USA) supplemented, when required, with erythromycin (5 μg mL−1), tetracycline (10 μg mL−1) and kanamycin (50 μg mL−1). For biofilm assays Tryptic Soy Broth (TSB) (EMD Millipore, Billerica, MA, USA), sterilized by filtration (autoclaving was found to reduce biofilm formation by B. cereus 905), was used. TSB broth was supplemented, where indicated, with 1% glucose, proteinase K (0.1 mg mL−1) (Omega Bio-Tek, Norcross, GA, USA) or DNaseI (5 U mL−1) (Qiagen, Valencia, CA, USA). Other media utilized were TSB without glucose (Peptone from casein [BD] 17 g L−1, Peptone from soymeal (Amresco, Solon, OH, USA) 3 g L−1, NaCl (Sigma, St. Louis, MO, USA) 5 g L−1 and dipotassium hydrogen phosphate (Macron, Center Valley, PA, USA) 2.5 g L−1), BHI (EMD Millipore), MSgg (Branda et al. 2001 (link)), LBGM (Shemesh and Chai 2013 (link)) and M9 (De Kievit et al. 2001 (link)). Escherichia coli DH5α was grown at 37°C in LB medium and on LB agar supplemented, when required, with ampicillin (100 μg mL−1), kanamycin (50 μg mL−1) or tetracycline (10 μg mL−1). The pH of biofilm cultures was measured by spotting 20–30 μL on pH-indicator strips (range 5.0–10.0) (EMD Millipore).
7
Antibiotic Susceptibility Testing of S. aureus
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S. aureus isolates obtained at C1 and C2 were suspended in sterile saline to 0.5 McFarland. Susceptibility testing was performed on Mueller-Hinton II agar 3.8% w/v (BD Diagnostic Systems, Sparks, MD, USA) using the standardized disk diffusion method in accordance with the European Committee on Antimicrobial Susceptibility Testing (www.eucast.org ) guidelines for the following antibiotics: cefoxitin (30 µg), fusidic acid (10 µg), erythromycin (15 µg), clindamycin (2 µg), rifampicin (5 µg), gentamicin (10 µg), trimethoprim-sulfamethoxazole (25 µg), tetracycline (30 µg), and norfloxacin (10 µg). All disks were from Oxoid, Basingstoke, UK.
8
Antibiotic Susceptibility Profiling of Bacterial Isolates
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Each isolate cultured from the spleen of cases submitted to the VDL (Fayetteville, AR) was individually grown under aerobic conditions at 37°C in 4 mL of BHI broth to achieve turbidity of 0.5 MacFarland. A cotton swab was submerged into the inoculated BHI broth and swabbed onto Mueller Hinton agar with 5% sheep blood (Catalog No. R01622, Remel, Lenexa, KS) to achieve a uniform lawn. The Mueller Hinton plate was then tamped with a spring-loaded antibiotic disc dispenser (BD, Catalog No. 260640, Sparks, MD) containing 12 different antibiotics: clindamycin (CC ), 2 μg/disc; erythromycin (E ), 15 μg/disc; florfenicol (FFC ), 30 μg/disc; gentamycin (GM ), 10 μg/disc; neomycin (N ), 30 μg/disc; penicillin (P ), 10 μg/disc; sulfadiazine (SD ), 0.25 μg/disc; spectinomycin (SPT ), 100 μg/disc; sulfamethoxazole/trimethoprim (SXT ), 23.75/1.25 μg/disc; oxytetracycline (T ), 30 μg/disc; tetracycline (TE ), 30 μg/disc; ceftiofur (XNL ), 30 μg/disc; (BD, Sparks, MD). The plate was incubated overnight at 37°C under aerobic conditions. After approximately 18 h, the zone of inhibition for each antibiotic was determined using a caliper, rounding to the nearest millimeter. Susceptibility was determined according to the American Standard of the Clinical and Laboratory Standards Institute (Hudzicki, 2009 ).
9
Antimicrobial Susceptibility Testing of CA-MRSA
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All S. aureus isolates were identified by mass spectrometry (MALDI-TOF, Biotyper, Bruker Daltonic GmBH, Bremen, Germany). The susceptibility of CA-MRSA isolates was tested against 16 antimicrobial agents using the disk diffusion method according to the guidelines of the Clinical Laboratory Standard Institute (CLSI) (13 ). The antibiotics tested were penicillin, cefoxitin, vancomycin, gentamicin, tobramycin, kanamycin, erythromycin, clindamycin, tetracycline, ciprofloxacin, trimethoprimsulfamethoxazole, chloramphenicol, rifampin, linezolid, mupirocin and fusidic acid (BD, Maryland, USA). Minimal inhibitory concentration (MIC) determination of oxacillin was performed using the E-test (bioMerieux, Marcy I’Etoile, France).
10
Antibiotic Susceptibility Profiling of MRSA
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The susceptibility patterns of the MRSA isolates were performed by the agar disk diffusion method according to the guidelines of the European Committee on Antimicrobial Susceptibility Testing (EUCAST) (15 , 16 ). The antibiotics tested were penicillin, cefoxitin, gentamicin, tobramycin, kanamycin, erythromycin, clindamycin, tetracycline, ciprofloxacin, trimethoprim-sulfamethoxazole, chloramphenicol, rifampin, linezolid, mupirocin and fusidic acid (BD, Sparks, USA). Minimal inhibitory concentration (MIC) determination of oxacillin, cefoxitin and vancomycin was performed using the E-test (bioMerieux, Marcy-l’Etoile, France).
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