Mb-NO analysis was carried out in accordance with the previous method (Gao et al., 2014 ) with minor modifications. The minced sample (10 g) was homogenized in 90 mL of phosphate buffer (pH 6.0, 0.01 mol/L) using a BagMixer (400CC; Interscience, Saint-Nom la Bretèche, France) for 90 s. Then the mixture was placed in a dark room at 4°C for 1 h and filtered with a nitrocellulose membrane (0.22 mm). The absorbance spectra of pigment extracts from the sausages were evaluated from 350 nm to 700 nm using a UV–Vis spectrophotometer (TU-1810; Beijing Purkinje General Instrument Co., Ltd., Beijing, China). For lipid oxidation, the minced sample (10 g) was homogenized in 50 mL of trichloroacetic acid solution (7.5% of trichloroacetic acid, 0.1% of EDTA, w/v) using a BagMixer (400CC; Interscience, Saint-Nom la Bretèche, France) for 90 s. Then the lipid oxidation was measured by the thiobarbituric acid (TBA ) method accordingly as described previously by measuring the absorbance of the reaction products from the oxidation products of unsaturated fatty acids and TBA (Racanicci et al., 2008 ).
Bagmixer
Manufactured by Interscience
Sourced in France, United States, Japan, United Kingdom
The BagMixer is a laboratory equipment used for homogenizing samples. It mixes and homogenizes the contents of a bag in a controlled and standardized manner.
Automatically generated - may contain errors
Lab products found in correlation
Ringer solution, by Merck Group (3 mentions)
Buffered peptone water (bpw), by Thermo Fisher Scientific (3 mentions)
Dilumat, by bioMérieux (2 mentions)
Mrs agar, by Thermo Fisher Scientific (2 mentions)
Bagfilter 400, by Interscience (2 mentions)
Bile salt, by Thermo Fisher Scientific (1 mentions)
Tryptic soy broth (tsb), by Thermo Fisher Scientific (1 mentions)
Novobiocin, by Merck Group (1 mentions)
Tryptic soy broth (tsb), by Merck Group (1 mentions)
Violet red bile dextrose agar, by Merck Group (1 mentions)
Vitek 2 compact system, by bioMérieux (1 mentions)
Porcine stomach mucin type 2, by Merck Group (1 mentions)
Bacteriological agar, by Merck Group (1 mentions)
Potato dextrose agar (pda), by Thermo Fisher Scientific (1 mentions)
Ciprofloxacin, by Merck Group (1 mentions)
Cm0509, by Thermo Fisher Scientific (1 mentions)
Maldi tof, by Bruker (1 mentions)
Cm0129, by Thermo Fisher Scientific (1 mentions)
Fastdna spin kit for soil, by MP Biomedicals (1 mentions)
Red blood cell (rbc), by Merck Group (1 mentions)
66 protocols using bagmixer
1
Nitric Oxide and Lipid Oxidation Analysis
2
Isolation and Confirmation of E. coli O157:H7 from Salad Samples
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Salad samples (25 g) were added to 225 mL TSB (Oxoid) supplemented with 20 mg L−1 of novobiocin (Sigma, MO, US) and 1.12 g L−1 of bile salts (Oxoid) as reported by Fusco et al. [32 (link)] and homogenized in a stomacher (Bag Mixer, Interscience) for 2 min. Triplicate series of tubes containing decimal serial dilution (from 10 to 10−5 grams of homogenate) were incubated for 48 h at 37°C. Turbid cultures were considered to be presumptive positive. Confirmation of presumptive E. coli O157:H7 was carried out by spread-plating 100 μL samples from each turbid tube onto tellurite-cefixime Sorbitol MacConkey agar plates (Becton Dickinson, Sparks, MD) supplemented with 200 μg mL−1 ampicillin. Plates were incubated overnight at 37°C and colonies showing the typical morphology (colorless or neutral/gray with a smoky center and 1-2 mm in diameter) were considered positive. For further confirmation, a portion of each typical colony was picked and tested using RIM E. coli O157:H7 latex agglutination assay (Remel Inc., Lenexa, KS).
3
Salmonella Growth Kinetics in Egg Whites
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Fresh shell eggs were purchased from a local market located in Jeonju, South Korea, and pasteurized LEW was purchased from an online retail market in South Korea. Fresh shell eggs were separated into egg yolk and egg white (EW) in a sterile biosafety cabinet (Esco Micro Pte. Ltd, Singapore, Singapore). The collected EW (stored in sterile sampling bags) (Merck, Darmstadt, Germany) were homogenized by a stomacher (BagMixer, Interscience, St. Nom, France) to ensure the consistency of the samples. Homogenized EW were subdivided into 10 mL aliquots in sterile conical tubes (SPL, Pocheon, Korea), and pasteurized LEW were also prepared in the same way. The sample tubes were prepared to fulfill the number of data points for each temperature.
To determine the mathematical models of Salmonella spp. in EW and LEW, six strains of Salmonella serovars were used in the experiments. S. Enteritidis (ATCC 13076, NCCP 14546), S. Typhimurium (NCCP 16207, NCCP 12219), S. Montevideo (NCCP 10140), and S. Kentucky (NCCP 11686) were provided by the American Type Culture Collection (ATCC) and the National Culture Collection for Pathogens (NCCP). All strains were individually inoculated with 10 mL Tryptic Soy Broth (Merck, Darmstadt, Germany), cultured at 37 °C, and shaken (140 rpm) overnight. The cultures were combined to make a cocktail of bacteria, which was used to inoculate the samples.
To determine the mathematical models of Salmonella spp. in EW and LEW, six strains of Salmonella serovars were used in the experiments. S. Enteritidis (ATCC 13076, NCCP 14546), S. Typhimurium (NCCP 16207, NCCP 12219), S. Montevideo (NCCP 10140), and S. Kentucky (NCCP 11686) were provided by the American Type Culture Collection (ATCC) and the National Culture Collection for Pathogens (NCCP). All strains were individually inoculated with 10 mL Tryptic Soy Broth (Merck, Darmstadt, Germany), cultured at 37 °C, and shaken (140 rpm) overnight. The cultures were combined to make a cocktail of bacteria, which was used to inoculate the samples.
4
Quantifying Spoilage Bacteria in Fish
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In order to determine the microbial load in the tested fish samples, 10 g of representative fish sample was homogenized with 90 mL of sterilized Ringer solution (Merck, Darmstadt, Germany) in an appropriate sterile stomacher bag for 60 s using a Stomacher (BagMixer®, interscience, St Nom la Bretèche, France). Then, 0.1 mL of 10-fold serial dilutions of fish homogenates were transferred and spread on the surface of appropriate culture media in Petri dishes for spoilage bacteria enumeration. Plate count agar (PCA, Merck, Darmstadt, Germany) was used for the enumeration of total viable count (incubation at 25 °C for 72 h). For the enumeration of Pseudomonas spp., cetrimide–fucidin–cephaloridine (CFC) agar (Merck, Darmstadt, Germany) was used (incubation at 25 °C for 48 h). Violet Red Bile Dextrose agar (VRBD, Merck, Darmstadt, Germany) was used for the enumeration of Enterobacteriaceae spp. using the pour plate method (incubation at 25 °C for 48 h).
Two replicates of at least three appropriate dilutions were enumerated. Microbial growth modeling was carried out using the Baranyi growth model [21 (link)], by fitting curves using the DMFit program (http://www.combase.cc/index.php/en/ ). Different kinetic parameters, i.e., the rate (k) of the microbial growth, lag phase (Lag), and the final microbial population (Nmax) were estimated at the tested processing conditions.
Two replicates of at least three appropriate dilutions were enumerated. Microbial growth modeling was carried out using the Baranyi growth model [21 (link)], by fitting curves using the DMFit program (
5
Isolation and Identification of Aeromonas Bacteria
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Ten grams of the samples were mixed with 90 mL sterile normal saline and flapped in BagMixer (Interscience, St. Nom, France) for 1 min. Then, the treated sample was continuously diluted and counted on Plate Count Agar (PCA, Aoboxing Bio-Tech, Beijing, China) and incubated at 28 °C for 48 h. After 48 h, all obviously distinct bacterial colonies were appraised and each colony was streaked onto Aeromonas Selective Medium Base (Ryan) followed by incubation at 28 °C for 24 h. The color, outline, size and other optical properties of the colonies were observed and recorded. The VITEK-2 Compact system (BioMerieux, Marcy l’Etoile, France) was used to identify the single colony [23 ]. Further study on the colony was validated with 16S rRNA PCR using the forward primer 27F (5′-AGAGTTTGATCCTGGCTCAG-3′) and the reverse primer 1492R (5′-ACGGCTACCTTGTTACGACTT-3′) amplify the 16S rRNA gene [24 (link)]. The gene sequences were analyzed by comparing the closely related sequences by BLASTN program in the GenBank database and phylogenetic analyses using MEGA 5.0 software.
6
Listeria monocytogenes detection in cheese
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The L. monocytogenes strains were cultured in TSBYE at 37 °C for 24 h. The sliced cheese pieces were irradiated with ultraviolet light in a laminar airflow cabinet for 15 min to eliminate the background microflora. The cheese samples were tested for the presence of L. monocytogenes before the experiments. Each sample was inoculated with 50 µL aliquots of the cocktail of L. monocytogenes strains to obtain a final concentration of approximately 4.5 log cfu/g. The inocula were evenly spread over the surface of the cheese samples and allowed to dry for 15 min for the bacterial attachment. The cheese samples were coated as described in Section 3.6 , placed separately in a sterile bag (Whirl-Pak; Nasco, WI, USA), and sealed. A total of 120 cheese samples (five treatments × three replicates × eight days) were prepared on different days. Each treatment was stored at 5 °C for 24 days and sampled at regular time-intervals for the analysis. The sample was homogenized with sterile 0.1% (w/v) peptone water using a stomacher (BagMixer®; Interscience, France) for 1 min. Appropriate serially diluted samples were spread onto Oxford Agar plates which were incubated at 37 °C for 24 h. The population of L. monocytogenes was expressed as log cfu/g samples.
7
Microbial Analysis of Processed Fish
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Aliquots of 25 g sectioned from muscle tissue were aseptically collected and introduced in stomacher bags (BagMixer®, Interscience, Puycapel, Cantal, France) adding 225 mL of maximum recovery diluent (MDR, Oxoid Ltd., UK). These samples were successively stomached for two minutes at room temperature to produce 10-fold dilutions. All incubated specimens were plated onto plate count agar (PCA, Thermo Scientific™ Oxoid Standard Plate Count Agar) and the obtained quantitative data were expressed in log cfu/g. The microbiological analysis also included the detection of pathogenic bacteria which are also reported in the EU Reg No. 2073/2005, and other bacteria more frequently identified in processed fish products. Selective culture media were performed following standardized methods, as reported in Table 3 .
8
Adhesion of ETEC H10407 to Mucin-Agar
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Adhesion experiments were adapted from Tsilia et al. (2016) as previously described92 (link),93 (link). Briefly, mucin-agar consisted of 5% porcine stomach mucin-type II (Sigma-Aldrich, St. Louis, MO, USA) and 1% bacteriological agar (Sigma-Aldrich, St. Louis, MO, USA), with a pH adjusted to 6.8 to mimic human small intestinal pH. Six-well plates containing mucin-agar were inoculated with ETEC strain H10407 (initial concentration of 107 CFU.ml−1). After 1-hour incubation (37 °C, 120 rpm), each well was rinsed twice with phosphate buffer saline (PBS) to remove non-adherent bacteria. Separation of adhered bacteria was mechanically performed by transferring aseptically the whole mucin layer into a sterile bag containing PBS and further homogenization in a 400 P BagMixer® for 10 min (Interscience, Bread, Netherlands). Adhered ETEC bacteria were quantified by plating onto LB agar. Experiments were performed in triplicate and agar without mucin was used as a negative control.
9
Microbiological Enumeration of Lactobacilli
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For microbiological enumeration, three pieces of each treatment were weighted, diluted (1 : 10) with saline solution (NaCl 8.6 g L−1), and homogenized in a blender (Bag Mixer, Interscience, Saint-Nom-la-Bretèche, France) for 2 minutes. Then, samples were submitted to tenfold serial dilution. L. plantarum and L. fermentum concentration was determined by plating on MRS agar after incubation at 30°C for 48 h. Mesophilic microorganism and yeasts and moulds were enumerated by plate counting on PCA or PDA (Oxoid) added with chloramphenicol (100 mg L−1) and incubated at 25 and 30°C for 48 h, respectively.
10
Cranberry and Pomegranate Extraction
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The extraction method applied was an adaptation from the study of Yang, Jia and Zu (2016) [16 (link)]. In brief, cranberry and pomegranate fruits were blended for approximately 60 s at 25 °C in a stomacher apparatus (Bagmixer, InterScience, SaintNomlaBretèche, France). Twenty grams of each puree was mixed with 200 mL of water (10% w/v) at 60 °C in an Erlenmeyer flask with agitation for two hours.
A 0.45 mm filter paper under vacuum was used for the filtration of the above aqueous solutions and centrifugation of the filtrates took place at 5000× g (PALL, LifeSciences, Portsmouth, UK) for 20 min at room temperature. Evaporation of the supernatants until a final volume of 5 mL at 50 °C in an evaporator (Model R204B3, Senco Technology Ltd., Shanghai, China) was the next step. Finally, 5 mL of distilled water was added (initial concentration: 2 g/mL) to the previous concentrated solutions. The samples were kept at −4 °C.
A 0.45 mm filter paper under vacuum was used for the filtration of the above aqueous solutions and centrifugation of the filtrates took place at 5000× g (PALL, LifeSciences, Portsmouth, UK) for 20 min at room temperature. Evaporation of the supernatants until a final volume of 5 mL at 50 °C in an evaporator (Model R204B3, Senco Technology Ltd., Shanghai, China) was the next step. Finally, 5 mL of distilled water was added (initial concentration: 2 g/mL) to the previous concentrated solutions. The samples were kept at −4 °C.
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