For determination of the colony-forming units (CFU), the specimens were vortexed for 2 min in sterile tubes with 1 mL 0.9% sodium chloride and for 1 min in an ultrasonic bath on ice. This solution was serially diluted up to 1 : 104 in saline solution and plated on Columbia blood agar (CBA, aerobic bacteria and facultative anaerobic bacteria) or on yeast-cysteine-blood agar, respectively (HCB, anaerobic bacteria). The HCB plates were incubated for 7 days in anaerobic jars (Merck) at 37°C using BBL GasPak Anaerobic System Envelopes (Becton Dickinson, NJ, USA), the CBA plates under aerobic conditions with 5% CO2 for 2 days.
Bbl gas pak anaerobic system envelopes
Manufactured by BD
Sourced in United States
The BBL® Gas Pak anaerobic system envelopes are designed to create an anaerobic environment for the cultivation of anaerobic microorganisms. The envelopes contain chemical reagents that absorb oxygen and generate carbon dioxide, establishing an anaerobic atmosphere within the sealed container.
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Lab products found in correlation
4 protocols using bbl gas pak anaerobic system envelopes
1
Determination of Microbial Colony Counts
2
Anaerobic Fermentation of Lactobacilli
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Lactobacilli were routinely grown in an MRS broth (Merck) [18 (link)]. Overnight grown cells were washed twice in saline (0.85% NaCl solution), and 10% of the bacterial suspension (107 cfu mL−1) was used to inoculate modified MRS broth and agar media (pH 6.8) containing either 2% glucose, 2% xylose, 2% xylobiose, or 2% XOS. The anaerobic fermentations were performed in 100-mL glass bottles at 37 °C for 48 h (BBL® Gas Pak anaerobic system envelopes, Becton Dickinson, NJ, USA).
3
Culturing Lactobacillus Strains Anaerobically
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In this study, we used three strains of lactobacilli from the collection of the Department of General and Applied Microbiology at Sofia University (Sofia, Bulgaria): Lactobacillus plantarum S26, Lactobacillus brevis S27, and Lactobacillus sakei S16. The strains were cultured overnight (16–18 h) on MRS (de Mann Rogosa Sharpe broth, Merck, Darmstadt, Germany) media [18 (link)] at 37 °C and in limitation of oxygen (BBL® Gas Pak anaerobic system envelopes, Becton Dickinson, Franklin Lakes, NJ, USA).
4
Enumeration of Microbial Populations in Food Samples
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Each sample of 10 mL was mixed with 90 mL of 0.85% (wt/vol) sterile physiological saline, and homogenized in a Stomacher Lab-blender 400 Circulator (Seward, Worthing, UK) for 2 min. Suitable serial dilutions in the same diluent were prepared. Lactic acid bacteria were enumerated on de Man, Rogosa, and Sharpe agar and enterococci on kanamycin azide agar, both incubated anaerobically by means of anaerobic jars and BBL GasPak anaerobic system envelopes (Becton Dickinson, Franklin Lakes, NJ) at 37°C for 48 h. Potato dextrose agar, added with chloramphenicol (Sigma-Aldrich), incubated at 25°C for 72 h, was used for the enumeration of yeasts, and violet red bile glucose agar incubated at 37°C for 24 h, was used for Enterobacteriaceae population. Staphylococcus aureus was enumerated in Baird Parker medium supplemented with egg yolk tellurite emulsion incubated at 35°C for 48 h. Standard cultivation methods were carried out for Salmonella spp. and Listeria monocytogenes isolation as recommended by ISO 6579 (ISO, 2002) and ISO 11290-1 (ISO, 2004) procedures, respectively.
All the media were from Oxoid-Thermofisher (Rodano, Italy), except where otherwise specified.
All the media were from Oxoid-Thermofisher (Rodano, Italy), except where otherwise specified.
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