From cultures of L. monocytogenes strains obtained on Columbia Agar with 5% sheep blood (CAB, bioMérieux, Marcy-l’Étoile, France) suspensions of a density of 0.5 MacFarland standard (5.80 × 108 CFU × mL−1) were prepared in 3 mL of Mueller Hinton Broth (MHB, Becton Dickinson, Franklin Lakes, New Jersey, USA). For this purpose, the optical density for the sterile MHB (Mueller Hinton Broth, Becton Dickinson, Franklin Lakes, New Jersey, USA) medium was first established. A sterile swab was then collected from a single colony grown on Columbia Agar with 5% sheep blood (CAB, bioMérieux, Marcy-l’Étoile, France) and loaded into the MHB (Mueller Hinton Broth, Becton Dickinson, Franklin Lakes, New Jersey, USA) medium, followed by measurement of the optical density of the suspension and subsequent colonization of L. monocytogenes added if necessary. The optical density of the suspension was set at 0.5 + the optical density of the sterile MHB (Mueller Hinton Broth, Becton Dickinson, Franklin Lakes, New Jersey, USA). The measurements were made with a DEN-1B denitometer from Biogenet (Józefów, Poland).
Mueller hinton broth (mhb)
Manufactured by BD
Sourced in United States, France, Germany, Poland, Canada
Mueller-Hinton broth is a microbiological growth medium used for the cultivation and susceptibility testing of bacteria. It provides the necessary nutrients for the growth of a wide range of microorganisms.
Automatically generated - may contain errors
Lab products found in correlation
Mueller hinton agar (mha), by BD (31 mentions)
Tryptic soy broth (tsb), by BD (15 mentions)
Gentamicin, by Merck Group (8 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (8 mentions)
Agar, by BD (6 mentions)
Tryptic soy agar, by BD (6 mentions)
Skim milk, by BD (6 mentions)
Crystal violet, by Merck Group (6 mentions)
Vancomycin, by Merck Group (5 mentions)
Amikacin, by Merck Group (5 mentions)
Isovitalex, by BD (5 mentions)
E z n a bacterial rna kit, by Omega Bio-Tek (4 mentions)
Smart bacterial protein extraction solution, by iNtRON Biotechnology (4 mentions)
Ceftazidime, by Merck Group (4 mentions)
Ciprofloxacin, by Merck Group (4 mentions)
Sabouraud broth, by BD (4 mentions)
Etest, by bioMérieux (4 mentions)
Tryptic soy broth tsb, by BD (4 mentions)
Chloramphenicol, by Merck Group (4 mentions)
Oxacillin, by Merck Group (4 mentions)
240 protocols using mueller hinton broth (mhb)
1
Standardized Listeria monocytogenes Culture
2
Brucella Strain Growth Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The bacterial strains and plasmids used in this study are listed in Supplementary Table S1 . All bacteria were grown either on tryptic soy agar (TSA, Pronadisa) plates or in tryptic soy broth (TSB, Scharlau) or Mueller-Hinton broth (Becton Dickinson, Difco) at 37°C. Where indicated, growth media were supplemented with kanamycin (Km) at 50 mg/ml, nalidixic acid (Nal) at 25 mg/ml, ampicillin (Amp) at 100 mg/ml, and/or 5% sucrose. Bacterial growth rates were determined at 37°C in Mueller–Hinton broth (Becton Dickinson, Difco), using a Bioscreen C apparatus (Lab Systems). All strains were stored in skim milk at -80°C. Work with Brucella was performed at the Biosafety Level 3 (BSL-3) laboratory facilities of the “Centro de Investigación Médica Aplicada de la Universidad de Navarra” (CIMA) and “Centro de Investigación y Tecnología Agroalimentaria de Aragón” (CITA), Spain.
3
Comparative Colistin Susceptibility Profiling
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Colistin MIC was determined for isolates AB1-5 with Etest (bioMérieux, Marcy l’Etoile, France), broth dilution and agar dilution. Colistin Etest was applied on Mueller-Hinton Agar (MHA) (Becton Dickinson, Franklin Lakes, NJ, USA) inoculated with a bacterial suspension of 0.5 McFarland in 0.85 % NaCl and was read after 24 h incubation at 36 °C. Broth dilution MIC of Colistin sulphate (Sigma-Aldrich corp., St Louis, MO, USA) was determined with 2-fold dilutions (0.016–256 mg/L) in glass tubes containing 1 mL Mueller-Hinton Broth (MHB) (Becton Dickinson), with an inoculum of approximately 1 × 105 CFU/mL. Broth dilution MIC was defined as the lowest concentration inhibiting visible growth after 24 h of incubation at 36 °C. Agar dilution MIC was determined with MHA (Becton Dickinson) plates with 2-fold dilutions of Colistin sulphate (Sigma Aldrich), ranging from 0.08 to 2.0 mg/L. A bacterial suspension of approximately 105 CFU/mL, determined by viable count, was prepared by dilution with NaCl 0.85 % of an overnight culture of 3 mL MHB (Becton Dickinson). Plates were spot inoculated, with two spots and 104 CFU in each spot. The plates were incubated overnight at 36 °C. Agar-MIC was defined as the lowest concentration that inhibited visible growth. All tests were performed in duplicates to ensure reproducibility.
4
Growth Conditions for Diverse Microorganisms
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The bacterial strains used in this study are shown in Table 1 . Escherichia coli and Pseudomonas aeruginosa were grown in Muller-Hinton broth (MHB, Becton Dickinson, Sparks, MD, USA) supplemented with 50 μg mL−1 of CaCl2 and 25 μg mL−1 of MgCl2, and methicillin-resistant S. aureus (MRSA) strains were grown in MHB supplemented with 25 μg mL−1 of CaCl2, 12.5 μg mL−1 of MgCl2, and 2% NaCl [24 ]. Streptococcus spp. were grown in brain-heart infusion (BHI, Becton Dickinson) anaerobically. C. albicans was grown in Sabouraud dextrose medium composed of 10 g L−1 of peptone and 40 g L−1 of glucose aerobically. F. nucleatum and P. gingivalis were grown in BHI supplemented with 5 μg mL−1 of hemin and 0.5 μg mL−1 of menadione anaerobically. Aggregatibacter actinomycetemcomitans was grown in Todd Hewitt Broth (OXOID Ltd., Hampshire, UK) anaerobically. For the biofilm formation assays of F. nucleatum, S. mutans, and MRSA T31, Trypticase soy broth (TSB, Becton Dickinson) supplemented with 5 μg mL−1 of hemin and 0.5 μg mL−1 of menadione, TSB supplemented with 0.3% sucrose, and TSB supplemented with 0.3% glucose were used, respectively. For biofilm formation assay of C. albicans, yeast nitrogen base medium at pH 7 containing 2.5 mmol L−1 of N-acetylglucosamine (YNBNP) [25 (link)] was used.
5
Determining Ceftiofur MIC in E. coli
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The minimum inhibitory concentration (MIC) of ceftiofur for each E. coli isolate was determined using a broth microdilution method and following Clinical and Laboratory Standards Institute standards [23 ]. Isolates were grown overnight on blood agar. A single colony was inoculated into 3 ml of sterile phosphate-buffered saline and brought to a 0.5 McFarland Standard, then 10 μl of this Standard were further diluted in 990 μl sterile phosphate-buffered saline. This final bacterial suspension (50 μl) was inoculated into 50 μl of two-fold serial dilutions of ceftiofur (USP, Rockville, MD) in Mueller Hinton broth (BD, Sparks, MD) ranging in concentration from 0.03 to 32 mg/L. Each bacterial suspension (50 μl) was also inoculated into a control well containing 50 μl Mueller Hinton broth (BD, Sparks, MD), with no antibiotic. The 96-well plates were incubated 18 hours at 37°C. The drug concentration in the first well with no visible growth was determined to be the MIC.
6
Antibiotic Minimum Inhibitory Concentration Determination
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The minimum inhibitory concentration (MIC) of all antibiotics was determined using the broth-microdilution method, according to the Clinical and Laboratory Standard Institute (CLSI) guidelines [13 ,14 ]. Briefly, cation-adjusted Mueller-Hinton broth (Becton, Dickinson and Co.; Franklin Lakes, NJ, USA) containing graded concentrations of antibiotics was freshly prepared. Study isolates that grew to an optic density of 0.5 McFarland units were diluted to 5 ×106 CFU/ml. Then 100 μl of antibiotic solution and 10 μl of bacteria suspension were added simultaneously to 96-well U-bottom microplates. After mixing with a vortexer, the microplates were incubated for 24 h at 37°C in ambient air. All susceptibility tests were performed in 3 independent experiments on different days.
7
Antibiotic Susceptibility of Acinetobacter baumannii
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Acinetobacter baumannii AB1845 and AB2092 were isolated from the sputum of the patient before and after cefoperazone/sulbactam therapy. Both isolates were stored at -80°C. The MIC was determined in cation-adjusted Mueller-Hinton broth (Becton-Dickinson, Sparks, MD, USA) using a broth microdilution according to Clinical and Laboratory Standards Institute (CLSI) standards. The MICs of the β-lactam/β-lactamase inhibitor combination, including cefoperazone/sulbactam, carbapenems, colistin, and polymyxin B, were determined.
8
Quantification of Tigecycline in Plasma
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Tigecycline reference standard (99% purity, Lot L910S56 and LN70S134) was purchased from Beijing J&K Scientific Ltd., and internal standard (IS) tigecycline-d9 (95% purity, Lot 2-NYL-17-1-PFZ) was purchased from Toronto Research Chemical, Inc., HPLC-grade formic acid and methanol were purchased from Thermo Fisher Scientific. Ultrapure water was prepared using a Milli-Q water purification device (Millipore, Bedford, MA, United States). Drug-free plasmas were obtained from the Department of Blood Transfusion, PLA General Hospital. A total of 134 AB isolates were used for susceptibility tests. Quality control strain ATCC25922 was purchased from National Institutes for Food and Drug Control, China. Mueller–Hinton Agar and Mueller–Hinton Broth were purchased from Becton, Dickinson and Company. Chromatographic analysis was performed using the Agilent 1260 high-performance liquid chromatography system (Agilent Technologies Inc.), and mass spectral analysis was performed using the Agilent 6460A mass spectrometer (Agilent Technologies Inc.) with an Agilent MassHunter Workstation B.06.00 software for data processing.
9
Bacterial Growth Kinetics Assay
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The overnight cultures were adjusted to OD600nm 0.05–0.1 in cation-adjusted Mueller Hinton broth ( Becton Dickinson and Company, Sparks, MD, USA), and 200 µL of each were added to each well in a 96-well polystyrene microplate. The OD600nm measurements were taken every 15 min with 5 s of agitation before and after the reading using the SpectraMax M5 spectrophotometer (Molecular Devices, San Jose, CA, USA) at 37 °C for 10 h. The assay was performed in five replicates. The measurements were plotted on a growth curve, and the doubling times were calculated using linear regression of the log-phase. The statistical significance was calculated using the Student’s t-test and ANOVA (p < 0.05).
10
Characterization of Staphylococcus Strain Gradients
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The strains employed in this study included methicillin-susceptible strain SH1000 [63 (link)] and a TTO-selected SCV, SH1000-TTORS-1 [30 (link)]. Mueller Hinton broth (MHB), Luria broth (LB), and bacteriological-grade agar were purchased from Becton Dickinson and Company (Franklin Lakes, NJ). All chemicals were purchased from Sigma Aldrich Co. (St. Louis, MO), unless otherwise noted. TTO utilized for this study was purchased from Aura Cacia (Urbana IA, Melaleuca alternifolia product code A191139) and contained 44.1% terpinen-4-ol and 3.9% α-terpineol (v/v) which complies with the TTO International Standard (ISO 4730).
Gradient plate analysis was performed with MHB overnight cultures and Mueller Hinton agar (MHA) as previously described [64 (link)], and confluent growth along the gradient was measured in mm ± standard deviation (n = 3). Colony size determination was carried out as previously described [29 (link)] by diluting overnight MHB cultures and plating on MHA and MHA containing 0.1% (v/v) Tween 80. Random colonies were then measured for each strain grown on both media using a caliper in mm ± standard deviation (n = 10).
Gradient plate analysis was performed with MHB overnight cultures and Mueller Hinton agar (MHA) as previously described [64 (link)], and confluent growth along the gradient was measured in mm ± standard deviation (n = 3). Colony size determination was carried out as previously described [29 (link)] by diluting overnight MHB cultures and plating on MHA and MHA containing 0.1% (v/v) Tween 80. Random colonies were then measured for each strain grown on both media using a caliper in mm ± standard deviation (n = 10).
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!