The automatized bacterial identification was made according to previously reported by our research group. Briefly, bacterial suspensions were made, by depositing two or three medium-sized colonies (2 to 3 mm) in BBL™ Crystal™ Inoculum Broth (Becton Dickinson, Franklin Lakes, NJ, USA). Said inoculum was adjusted to a Mac Farland 1.0 scale (Expected CFU/mL 3.0 × 108). The inoculum was deposited in BBL™ Crystal™ Enteric/Nonfermenter or Anaerobe ID Kit plates, incubated at 37 °C for 18 h, without CO2 and 40–60% humidity. Finally, the plates were read by the BBL™ Crystal™ AutoReader (Becton Dickinson, Franklin Lakes, NJ, USA) and the results were analyzed with the BBL™ Crystal™ MIND v.5.05 Software (Becton Dickinson, Franklin Lakes, NJ, USA) [26 (link)].
Bbl crystal inoculum broth
Manufactured by BD
Sourced in United States
BBL Crystal Inoculum Broth is a sterile, aqueous medium used for the preparation of standardized inocula for use in microbiological testing procedures. The broth provides a consistent suspension of microorganisms for inoculation purposes.
Automatically generated - may contain errors
Lab products found in correlation
Bbl crystal autoreader, by BD (2 mentions)
Ampicillin, by BD (2 mentions)
Cefepime, by BD (2 mentions)
Cefazolin, by BD (2 mentions)
Mezlocillin, by BD (2 mentions)
Carbenicillin, by BD (2 mentions)
Cefaclor, by BD (2 mentions)
Cefoperazone, by BD (2 mentions)
Cefotetan, by BD (2 mentions)
Ampicillin sulbactam, by BD (2 mentions)
Piperacillin tazobactam, by BD (2 mentions)
4 protocols using bbl crystal inoculum broth
1
Automated Bacterial Identification Protocol
2
Antimicrobial Susceptibility Testing Protocol
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The antimicrobial susceptibility testing was done following a previously reported method [26 (link)]. Briefly, the pure colonies obtained from blood agar were resuspended in bacterial suspensions, and were made depositing two or three medium-sized colonies (2 to 3 mm) in BBL Crystal Inoculum Broth (Becton Dickinson; Franklin Lakes, NJ, USA). The obtained inoculum was adjusted while using a Mac Farland 0.5 reading (Expected CFU/mL 1.5 × 108), and cultured in Müeller Hinton 150 × 15 mm2 media BD BBL (Becton Dickinson Franklin Lakes, NJ, USA). The antibiotic discs were applied with the Sensi-Disc Designer Dispenser System. The antibiotics panel was conformed by ampicillin (10 µg), ampicillin/sulbactam (10/10 µg), mezlocillin (75 µg), carbenicillin (100 µg), piperacillin/tazobactam (100/10 µg), cefazolin (30 µg), cefaclor (30 µg), cefepime (30 µg), cefoperazone (75 µg), and cefotetan (30 µg) from Becton Dickinson (Franklin Lakes, NJ, USA), Müeller Hinton medium were incubated at 37 °C for 24 h in both aerobic and anaerobic conditions. In anaerobic conditions, the media were placed in an anaerobic jar (BD BBL™ GasPak™), with a C02 gas generators envelope (BD BBL™) and another envelope of BD BBL™ GasPak™ anaerobic indicator placed on them.
3
Antimicrobial Susceptibility Testing Protocol
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Antimicrobial susceptibility testing was carried out by the Kirby–Bauer method under the Clinical and Laboratory Standards Institute protocol (NCCLS standards). The pure colonies obtained from blood agar were resuspended in bacterial suspensions were made depositing two or three medium-sized colonies (2 to 3 mm) in BBL Crystal Inoculum Broth (Becton Dickinson; Franklin Lakes, NJ, USA). The obtained inoculum was adjusted while using a Mac Farland 0.5 reading (Expected CFU/mL 1.5 × 108), and cultured in Müeller Hinton 150 × 15 mm2 media BD BBL (Becton Dickinson Franklin Lakes, NJ, USA). The antibiotic discs were applied with the Sensi-Disc Designer Dispenser System. The antibiotics panel was conformed by ampicillin (10 µg), ampicillin/sulbactam (10/10 µg), mezlocillin (75 µg), carbenicillin (100 µg), piperacillin/tazobactam (100/10 µg), cefazolin (30 µg), cefaclor (30 µg), cefepime (30 µg), cefoperazone (75 µg), and cefotetan (30 µg) from Becton Dickinson (Franklin Lakes, NJ, USA)
4
Bacterial Identification Using BBL Crystal System
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The bacterial identification was made from the pure colonies that were obtained from blood agar using Gram staining and oxidase, Indol and biochemistry tests. The BD BBL Gram Stain Kit (Becton Dickinson; Franklin Lakes, NJ, USA) was used. Oxidase and Indol tests were carried out by using BBL DrySlide Oxidase kit (Becton Dickinson, Franklin Lakes, NJ, USA) and BBL DrySlide Indole kit (Becton Dickinson; Franklin Lakes, NJ, USA) according to the manufacturer’s specifications. The bacterial identification was performed using the BBL Crystal Enteric/Nonfermenter ID system (Becton Dickinson; Franklin Lakes, NJ, USA). Briefly, bacterial suspensions were made depositing two or three medium-sized colonies (2 to 3 mm) in BBL Crystal Inoculum Broth (Becton Dickinson; Franklin Lakes, NJ, USA). That inoculum was adjusted to a Mac Farland 1.0 scale (Expected CFU/mL 3.0 × 108). To adjust the scale, a CrystalSpec Nephelometer (Becton Dickinson; Franklin Lakes, NJ, USA) was used. The inoculum was deposited in BBL Crystal Enteric/Nonfermenter ID Kit plates, incubated at 37 °C for 18 h, without CO2 and 40 to 60% humidity. Finally, the plates were read using a BBL Crystal AutoReader (Becton Dickinson; Franklin Lakes, NJ, USA) and the results were analyzed with the BBL Crystal MIND Software (Becton Dickinson; Franklin Lakes, NJ, USA).
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