The bacterial strains and media used in this study are described in Table 1 . All bacterial strains were cultured at 37°C for 24 h. All media were purchased from BD (USA), and the agar medium was prepared with 1.8% (w/v) Bacto Agar (BD). Four samples (chicken breast and three animal byproducts) were collected (200 g for each sample) from the Garak market (Korea) to isolate the foodborne pathogens (Table 1 ). After sample collection, the collected samples were cut using a sterile scalpel. Twenty-five grams of collected samples was transferred into 3M sterilized bag (USA) and suspended with 225 ml of sterilized phosphate-buffered saline (PBS) for homogenization. Homogenization was performed using BagMixer 400 (Interscience, France) with speed of 4 m/s for 30 s. After homogenization, samples were serially diluted up to 10−5, spread onto the selective agar plate specific for each pathogen (Table 1 ), and incubated as previously described. After incubation, a single colony was collected and streaked on fresh culture medium agar plate (Table 1 ). The selected bacterium was identified using 16S rRNA gene sequencing. In particular, pathogenic E. coli was further identified using pathogen type-specific gene PCR. The identified bacterium was stored at −80°C in 20% (w/v) sterilized glycerol solution.
Bagmixer 400
Manufactured by Interscience
Sourced in France, United Kingdom
The BagMixer 400 is a laboratory equipment designed for the homogenization of samples. It is used to mix and blend various types of samples, such as food, environmental, and medical samples, in a sealed bag. The device employs a reciprocating motion to ensure thorough mixing of the sample.
Automatically generated - may contain errors
Lab products found in correlation
Ringer s solution, by Merck Group (4 mentions)
Anaerocult a, by Merck Group (3 mentions)
Optizen 2120uv, by Mecasys (3 mentions)
Petrifilm aerobic count plate, by 3M (2 mentions)
Cycloheximide (chx), by Merck Group (2 mentions)
Mrs agar, by Thermo Fisher Scientific (2 mentions)
Egg yolk tellurite emulsion, by Thermo Fisher Scientific (2 mentions)
Baird parker agar, by Thermo Fisher Scientific (2 mentions)
Ringer solution strength, by Lab M (2 mentions)
Oxford listeria selective agar, by Merck Group (2 mentions)
Mrs medium, by Thermo Fisher Scientific (2 mentions)
Ph basic 20, by Crison (2 mentions)
Buffered peptone water (bpw), by Thermo Fisher Scientific (2 mentions)
Fraser broth, by BD (2 mentions)
Bacto agar, by BD (1 mentions)
Spd1010, by Thermo Fisher Scientific (1 mentions)
Amicon ultra 15 centrifugal filter unit, by Merck Group (1 mentions)
Mrs agar medium, by Thermo Fisher Scientific (1 mentions)
Peptone, by Merck Group (1 mentions)
Peptone water, by Merck Group (1 mentions)
93 protocols using bagmixer 400
1
Isolation and Identification of Foodborne Pathogens
2
Peptide Extraction from Samples
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Peptide extraction was conducted using a modified version of the method by Gallego et al. (2016) . Ten grams of sample were homogenized with 50 mL of 0.01 N HCl for 3 min in a stomacher (BagMixer 400, Interscience, Saint Nom, France). The homogenate was centrifuged at 10,000 × g for 30 min at 4°C and filtered through glass wool. Three volumes of ethanol were mixed with the filtrate, and it was stored for 24 h at 4°C. The mixture was centrifuged at 10,000 × g for 30 min at 4°C, and the supernatant was lyophilized using a vacuum evaporator (SPD1010, Thermo Fisher Scientific Inc). Lyophilized samples were dissolved in 5 mL of 0.01 N HCl, neutralized to pH 7.0 using NaOH, and filtered using a 0.45-µm nylon membrane filter (Millipore Corp., Bedford, MA). The filtrates were centrifuged in centrifugal filter-containing tubes (Amicon Ultra-15 Centrifugal Filter Unit, Millipore) at 10,000 × g for 30 min.
3
Microbial Profiling of Dairy Products
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Cell suspensions of milk samples were subjected to decimal serial dilutions in Ringer’s solution (1:10), while GPP, curd and cheese samples were first homogenized in Ringer’s solution by a stomacher (Bag-Mixer 400; Interscience, Saint Nom, France) for 2 min at the maximum speed (blending power 4) and then serially diluted. Cell suspensions of GPP were subjected to plate count for the main microbial groups belonging to the pro-technological, spoilage and pathogenic populations following the approach of Cruciata et al. [16 (link)].
Cell suspensions of raw milk and pasteurized milk were analyzed for total mesophilic microorganisms (TMM), mesophilic rods and cocci as reported by Barbaccia et al. [15 (link)].
Milk inoculated with each NMSC, curd and cheese samples were analyzed only for the levels of TMM and L. lactis on skim milk agar [17 ] incubated aerobically at 30 °C for 72 h and M17 agar incubated anaerobically at 30 °C for 48 h, respectively. All media and supplements were purchased from Biotec. Plate counts were performed in duplicate.
Cell suspensions of raw milk and pasteurized milk were analyzed for total mesophilic microorganisms (TMM), mesophilic rods and cocci as reported by Barbaccia et al. [15 (link)].
Milk inoculated with each NMSC, curd and cheese samples were analyzed only for the levels of TMM and L. lactis on skim milk agar [17 ] incubated aerobically at 30 °C for 72 h and M17 agar incubated anaerobically at 30 °C for 48 h, respectively. All media and supplements were purchased from Biotec. Plate counts were performed in duplicate.
4
Isolation and Enumeration of Mesophilic Lactic Acid Bacteria in Italian Dairy Products
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Samples of MBC were received on the day of production from four dairy factories located in different provinces of central and southern Italy (Latina-LT; Salerno-SA; Caserta-CE; Foggia-FG). We exclusively selected dairy plants with associated animal farming, which guarantees reproducible sources of milk and associated microbiota profiles for cheese production. Samples were stored at 4°C and processed within 12 h. Pooled or single samples of MBC were homogenized with a BagMixer400 (Interscience, France) in sodium citrate solution (2% w/v) at a concentration of 0.5 g/mL. In order to test the titer of mesophilic cultivable LAB, serial dilutions were made in Quarter Strength Ringer's solution and plated on MRS agar medium (Oxoid Ltd, Basingstoke, Hampshire, England), as previously reported [13 (link)]. Plates were incubated at 30°C for 48 h, under aerobic and anaerobic conditions (Anaerocult A, Merck, Germany).
5
Assessing Antimicrobial Efficiency of APPJ on Fresh-Cut Leaves
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The antimicrobial efficiency of APPJ treatment was assessed by measuring the total aerobic bacterial count of the fresh-cut leaf surface. To this purpose, leaves were homogenized in sterile 2% (w/v; pH 7.5 ± 0.1) dipotassium hydrogen phosphate solution (Sigma-Aldrich, St. Louis, MO, USA) for 2 min using a Stomacher blender (BagMixer 400, Interscience, St Nom la Bretèche, France). Ten-fold serial dilutions of each sample were prepared in quarter-strength Ringer’s solution (Scharlau Microbiology, Barcelona, Spain) and plated on a Petrifilm Aerobic Count plate (3M Minneapolis, MN, USA) according to the manufacturer’s instructions. The plates were incubated at 30 °C for 72 h. Each experiment was performed in triplicate. The results were expressed as log10 CFU per gram of product (log10 CFU/g, mean values ± standard error).
6
Biofilm Growth on Crab and Shrimp Surfaces
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The crab and shrimp surfaces (2 cm, 2 cm, and 0.1 cm, respectively) were treated as previously described [43 (link),44 (link)]. To eliminate any leftover substance, oil, and bacteria, the food samples were washed with distilled water (DW) and 70% ethanol. The mixed culture (100 µL) was placed in a 50 mL Falcon tube with a coupon in 10 mL TSB for biofilm growth. At different sub-MIC conditions (0, 1/2, 1/4, and 1/8 MIC), the cells were incubated for 24 h. After biofilm development, the samples were washed with DW to eliminate any detached adherent cells. After that, the samples were homogenized in 710 mL WhirlPak filter stomacher bags with 90 mL of 0.1% PW (Oxoid) using a stomacher (BagMixer 400; Interscience, Paris, France) at a maximum speed for 2 min. Then, the suspensions were cultured on PALCAM agar; cell counts were determined using the plate count method. Log CFU/cm2 was used to describe the biofilm cells.
7
Isolation and Enumeration of Acetic Acid Bacteria
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Apricot samples (100 g) were homogenized using a stomacher (Bagmixer 400, Interscience, France) and allowed to ferment with their natural microbiota in a sterile jar at 30℃ for 15 d for selective growth of AAB under an alcohol-rich and a low pH environment [7, 8] . Serial dilutions of the fermented apricot homogenates and vinegar samples were prepared in peptone water (0.1% [w/v], Merck KGaA, Germany) and were plated on acetic acid medium (AAM) agar (1% glucose [Sigma-Aldrich, USA], 0.5% ethanol [Isolab Laborgeräte GmbH, Germany], 0.3% acetic acid [Sigma-Aldrich], 1.5% peptone [Merck], 0.8% yeast extract [Biolife, Italy], and 1.5% agar [Merck], all w/v except acetic acid and ethanol, which were v/v) [9, 10] and glucose yeast extract calcium carbonate (GYC) agar (10% glucose [Sigma-Aldrich], 1.0% yeast extract [Biolife], 2.0% calcium carbonate [Sigma-Aldrich], and 1.5% agar [Merck], w/v) [11] . Both media were supplemented with 0.4% (w/v) cycloheximide (Sigma-Aldrich) to inhibit yeast growth [9] . The petri plates were incubated at 30℃ for 72 h.
Morphologically different colonies were selected from the plates with 25-250 colonies and purified by streaking onto other plates with the corresponding isolation medium. Isolates were grown in isolation medium broths containing 20% glycerol and kept at -80℃ for long-term storage.
Morphologically different colonies were selected from the plates with 25-250 colonies and purified by streaking onto other plates with the corresponding isolation medium. Isolates were grown in isolation medium broths containing 20% glycerol and kept at -80℃ for long-term storage.
8
Microbiological Analysis of Food Samples
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Microbial analysis was measured by homogenizing 10 g of sample from each replicate with 90 ml of sterile peptone water for 90 s using a stomacher blender (Bag Mixer 400; InterScience, France). Serial dilutions of fruit homogenates were prepared and 1 ml of final preparation was poured in plate count agar under sterile conditions (Valero et al., 2005 ). All plates were incubated for 5 days at 25°C and counts of colony forming were evaluated. Results were expressed as Log CFU/g.
9
Microbial Analysis of Fruit Samples
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Microbial analysis was performed by homogenizing 10 g of the sample from each replicate with 90 ml of sterile peptone water (Oxoid, Basingstoke, UK) for 90 s using a stomacher blender (Bag Mixer 400; InterScience, St.‐Nom‐La‐Bretèche, France). The blend was diluted 10 fold and 1 ml of the final preparation was poured in an agar plate (Mold Count Plates, ABRI, Karaj, Iran) under sterile conditions. All plates were incubated at 25°C for 5 days. The results were expressed as the logarithm of the number of colonies per 10 g of fruit FW (Valverde et al., 2005 ).
10
Microbial Analysis of Steak Surfaces
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Steak surface samples (~3 mm thick slices; 10 g) were taken aseptically, chopped, and transferred to sterile lateral filter bags (BagPage; Interscience, St Nom, France), with addition of 90 mL of sterile tryptone salt solution at 0.85% (w/v). Samples were mixed with a blender (BagMixer 400; Interscience, France) for 2 min at room temperature. A 10-fold dilution series was prepared to perform microbial analysis. Diluted sample solutions were cultured on plate count agar (Land-Bridge Co., Ltd., Beijing, China) and incubated at 37°C for 48 h. Results were expressed as log colony-forming unit (CFU)/g sample.
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