The grounded barley by‐products were obtained from local mill (Ustukiu malunas Ltd., Pasvalys, Lithuania) in 2018. Cereal by‐products samples were collected according to standard procedures described in ISO 24333:2009 (Cereals and cereal products—Sampling, ISO, 2009 :2009 ). Samples were collected during barley milling process and stored in the dark at ambient temperature until experiment. The P. acidilactici strain LUHS29, previously isolated from spontaneous fermented cereals and which showed broad antimicrobial activity against opportunistic and pathogenic strains (Bartkiene et al., 2019 ), was stored at −80°C and cultured at 30°C for 48 hr in MRS broth (CM0359, Oxoid Ltd) with the addition of 40 mmol/L fructose and 20 mmol/L maltose prior to use. The fermentation of barley by‐products was performed with a multiplied P. acidilactici strain (3% by volume of the pure P. acidilactici strain, diluted in MRS broth added to cereal/water mass). Three parallel replicates of fermented samples were prepared, and each fermented sample analysis was repeated three times. The water content of the end‐product for SSF was 450 g/kg; for SMF, it was 650 g/kg. Fermentation was carried out for 72 hr at 32 ± 2°C. Unfermented barley by‐products were used as a control.
Cm0359
Manufactured by Thermo Fisher Scientific
Sourced in United Kingdom
The CM0359 is a laboratory equipment product manufactured by Thermo Fisher Scientific. It is designed for use in scientific and research applications. The core function of the CM0359 is to provide a controlled environment for various laboratory processes.
Automatically generated - may contain errors
Lab products found in correlation
Microbank system, by Pro-Lab Diagnostics (5 mentions)
Cm0361, by Thermo Fisher Scientific (2 mentions)
Br 38, by Thermo Fisher Scientific (2 mentions)
Cm0325, by Thermo Fisher Scientific (1 mentions)
Cm0069, by Thermo Fisher Scientific (1 mentions)
Cm0469, by Thermo Fisher Scientific (1 mentions)
Mrs broth, by Thermo Fisher Scientific (1 mentions)
20 protocols using cm0359
1
Fermentation of Barley By-Products
2
Cecal Metabolomics of Conventional and GF Mice
3
Sourdough Fermentation Strains for BWP
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Five strains of LAB were used for the fermentation of BWP—Levilactobacillus brevis LUHS173, Liquorilactobacillus uvarum LUHS245, Lactiplantibacillus plantarum LUHS135, Lacticaseibacillus paracasei LUHS244 and Lacticaseibacillus casei LUHS210—which were isolated from spontaneous sourdough and provided from the collection of the Lithuanian University of Health Sciences [16 (link)]. Prior the use, LAB strains were maintained at −80 °C in a 25% glycerol solution in a Microbank system (Pro-Lab Diagnostics, Merseyside, UK) and before the fermentation strains were activated in MRS broth (de Man, Rogosa, and Sharpe, CM 0359, Oxoid Ltd., Hampshire, UK) for 48 h at 30 °C.
4
Bacterial Enumeration in Food Samples
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For bacterial enumeration, 1 g sample was transferred in 9 mL phosphate buffer saline (PBS) and serially diluted. The Lactobacillus spp., Escherichia coli, Salmonella spp., total coliform, and total viable bacteria (aerobes and anaerobes) were quantified on de Man Rogosa and Sharpe (CM0359, Oxoid UK), xylose lysine deoxycholate (CM0469, Oxoid UK), eosin methylene blue (CM0069, Oxoid UK), MaConkey’s (CM0505, Oxoid UK) and plate count agar (CM0325, Oxoid UK), respectively. The colonial counts were estimated through log cfu/g using spread plate technique (Andrews et al., 2014 ).
5
Cecal Metabolomics of Conventional and GF Mice
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Cecal contents from conventional or GF mice were collected and immediately placed in De Man, Rogosa, and Sharpe media (MRS broth – Oxoid, CM0359) in oxygen evacuated headspace vials. The control mouse diet was placed in MRS broth. Samples were incubated at 37°C and were collected over a 24h time period and prepared for metabolomics analysis as above.
6
Screening Lactobacillus Strains for Probiotics
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A total of 235 Lactobacillus isolates were tested using an antagonistic activity assay as described by Schillinger and Lucke, 1989 , Teo and Tan, 2005 , and the four strains of Lactobacillus isolates were selected as probiotic candidates by largest inhibition zone appearance with indicator pathogenic strains of C. perfringens and Escherichia coli. These four strains of Lactobacillus were tentatively identified as Lactobacillus johnsonii, Lactobacillus crispatus, Lactobacillus salivarius and one unidentified Lactobacillus sp.
All the strains were kept at −20°C in de Man, Rogosa, Sharpe (MRS) broth (Oxoid, CM0359) with 40% glycerol. The culture medium used for growth was MRS agar (Oxoid, CM0361). The overnight culture of each Lactobacillus isolate was used as a feed additive probiotic candidate after anaerobic incubation at 39°C for 24 h.
All the strains were kept at −20°C in de Man, Rogosa, Sharpe (MRS) broth (Oxoid, CM0359) with 40% glycerol. The culture medium used for growth was MRS agar (Oxoid, CM0361). The overnight culture of each Lactobacillus isolate was used as a feed additive probiotic candidate after anaerobic incubation at 39°C for 24 h.
7
Bacteria-host cell interaction protocol
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Bacteria from Danone collection (Table A in S1 File ) were cultivated in MRS (Man, Rogosa and Sharpe medium, Oxoid CM0359) at 37°C in pseudo-aerobic condition. Bacterial cultures (stationary phase) were centrifuged at 5,000x g for 10 min. Conditioned media (CM) were then collected, and filtered on 0.2μm PES filters. Bacterial pellets were washed twice in PBS and resuspended in antibiotic-free DMEM at OD600 = 0.1 (corresponding to mean Multiplicity Of Infection ranging from 23 to 113 bacteria for 1 cell) for bacteria. Cells were stimulated with 20% final volume of bacterial culture.
To respect the same ratio (bacteria/cell), Heat Inactivated (HI) bacteria, prepared at OD600 = 1 and heated at 80°c for 20 min, were added at 10% of final volume.
Conditionned media (CM), were used at 10% of final volume to limit the presence of lactic acid.
TranswellTM permeable support (Corning) was used to separate bacterial strain from cells for contact dependency test. In those assay, HT–29 were grown in the bottom of 24-well plates, transwell were then added and bacteria were seeded in the transwell preventing direct contact.
To respect the same ratio (bacteria/cell), Heat Inactivated (HI) bacteria, prepared at OD600 = 1 and heated at 80°c for 20 min, were added at 10% of final volume.
Conditionned media (CM), were used at 10% of final volume to limit the presence of lactic acid.
TranswellTM permeable support (Corning) was used to separate bacterial strain from cells for contact dependency test. In those assay, HT–29 were grown in the bottom of 24-well plates, transwell were then added and bacteria were seeded in the transwell preventing direct contact.
8
Extruded and Fermented Wheat Bran Characterization
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Wheat bran was obtained from the SME “Ustukiu malunas” (Pasvalys, Lithuania). Wheat bran samples (WB) were extruded at 130 °C, speed of the screw—25 rpm and fermented with L. uvarum LUHS245 strain. The LUHS245 strain, before the experiment, was stored at −80 °C in a Microbank system (Pro-Lab Diagnostics, Birkenhead, Wirral, UK) and grown in de Man, Rogosa and Sharpe (MRS) broth (CM 0359, Oxoid, Basingstoke, Hampshire, UK) at 30 °C for 48 h prior to use.
The following parameters for WB were established: pH, total titratable acidity (TTA), L(+) and D(-) lactic acid bacteria concentration, LAB, mould/yeast (M/Y), total bacteria (TBC), and total enterobacteria (TEC) counts; sugars concentration (fructose, glucose, sucrose, maltose); amino acids and biogenic amines concentration. Non-extruded and non-fermented WB samples were used as control.
The following parameters for WB were established: pH, total titratable acidity (TTA), L(+) and D(-) lactic acid bacteria concentration, LAB, mould/yeast (M/Y), total bacteria (TBC), and total enterobacteria (TEC) counts; sugars concentration (fructose, glucose, sucrose, maltose); amino acids and biogenic amines concentration. Non-extruded and non-fermented WB samples were used as control.
9
Fermentation of Pea Seeds using Lactobacillus
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Peas (Ps) were obtained from the “Galinta” enterprise (Kaunas, Lithuania) in 2018. Before treatments, Ps seeds were milled (particle size 2 mm) using a laboratory mill (Braun, Germany). The LAB strains L. casei LUHS210 and L. uvarum LUHS245 strains were obtained from the Lithuanian University of Health Sciences collection (Kaunas, Lithuania). Lactobacillus casei and L. uvarum strains were stored at −80°C in a Microbank system (Pro‐Lab Diagnostics, UK) and propagated in de Man–Rogosa–Sharpe (MRS) broth (CM 0,359, Oxoid Ltd, Hampshire, UK) at 30 ± 2°C for 48 hr, before their use for submerged fermentation (SMF) and SSF of Ps.
10
Isolation and Cultivation of LAB SR6 from Bali Cattle
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LAB SR6 isolate obtained from the colon of Bali cattle with a high tolerance to the environment was taken from a 30% glycerol stock, which was stored at −20°C. It was thawed at 4°C for 15 min, planted at 27°C in sterile De Man, Rogosa, and Sharpe (MRS) broth media (Oxoid, CM0359, England), and then incubated at 37°C for 24 h. The grown isolate was ready to be used for deoxyribonucleic acid (DNA) extraction and bacteriocin production [10 ].
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