Cell-free supernatants (CFS) of L. reuteri LR47 were prepared according to the method described by Lüthi-Peng et al. (31 (link)). A single colony with typical morphology (small pinpointed and creamy white colony) from the MRS agar plates was picked and inoculated into MRS broth and incubated anaerobically (gas pack system, HiMedia) at 37 °C for 20 h. After incubation, the culture was centrifuged (Precision Biotech Instruments Pvt. Ltd., Delhi, India) at 4000×g and 4 °C for 10 min. The supernatant was filter-sterilised through a 0.22-μm syringe filter (Millipore, New Delhi, India) to remove any remaining cells. The cell pellet was used for secondary fermentation to prepare a suspension of MRS supplemented with glycerol (360 mM). The supernatant obtained from secondary fermentation of MRS with glycerol was designated MRS-Gly, while the supernatant derived from MRS broth was designated MRSB. Finally, the pH values of the above supernatants were determined using a portable pH meter (Orion Star™ A111; Thermo Fisher Scientific, Waltham, MA, USA). The pH of the supernatants was adjusted from pH=4.6 to 6.5 (in order to eliminate the cause of microbial death due to acidic pH) using 0.1 M sodium hydroxide at room temperature prior to testing their antimicrobial activity.
Orion star a111
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Orion Star A111 is a benchtop pH meter designed for basic pH, mV, and temperature measurements. The device features an easy-to-read LCD display, automatic temperature compensation, and intuitive button controls. It is a reliable instrument for routine pH testing in various laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
Sytox green, by Thermo Fisher Scientific (3 mentions)
S230 sevencompact conductivity meter, by Mettler Toledo (2 mentions)
0.22 μm syringe filter, by Merck Group (1 mentions)
Gas pack system, by HiMedia (1 mentions)
Nutrient agar medium, by HiMedia (1 mentions)
Violet red bile agar, by HiMedia (1 mentions)
Phenol, by Merck Group (1 mentions)
Optima 3200 xl, by PerkinElmer (1 mentions)
Uvmini 1240, by Shimadzu (1 mentions)
Kinetic enzyme assay kit, by Salimetrics (1 mentions)
Ics 3000, by Thermo Fisher Scientific (1 mentions)
Dr 5000, by HACH (1 mentions)
19 protocols using orion star a111
1
Preparation of Cell-free Supernatants from Lactobacillus reuteri
2
Analytical Methods for Biomethanation Process
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The SCOD, TS, TSS (Total Suspended Solid), VS, VSS (Volatile Suspended Solid), and pH were analyzed according to the standard method (American Public Health Association (APHA), 1998). In brief, SCOD was detected by using potassium dichromate reagent and sulfuric acid reagent. Before testing, the sample was centrifuged for 5 min and filtered using the 0.45 µm membrane filter. TS and VS were measured after the sample was dried at 105°C and 550 °C in the oven (Precision Oven Model JA-72) and furnace (Model KEO-27L), respectively. The pH was determined by a pH meter (Thermo Scientific Orion Star A111). For the biogas composition was measured by gas chromatography (GC) with a Thermal Conductivity Detector (TCD) (GC System HP 6890 Series, USA). The temperature of the injector and detector were set at 175 °C and 250 °C, respectively. Helium (He) was used as the carrier gas at a flow rate of 40 mL/min. High-Performance Liquid Chromatography (HPLC-CM5000 series, Japan) from HITACHI was used to examine the volatile fatty acids (VFAs) concentration in the digester mixture.
3
Grape Ripeness Indicators Analysis
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In both years, all fruit were harvested from experimental vines and used for analysis of fruit total soluble solids (TSS), pH, and titratable acidity (TA), all of which are indicators of grape ripeness (Wolf, 2008 ). Yeast assimilable nitrogen (YAN) was also measured as it represents the pool of fruit N that can be utilized by yeast for alcoholic fermentation (Waterhouse et al., 2016 ). Clusters were harvested on September 27 in 2019 and September 30 in 2020, counted, and weighed, and a subsample of five clusters per vine were placed on dry ice and transported to the laboratory for storage at −80°C until analysis. One hundred berries per vine were randomly selected for juice chemistry analysis. The berries were weighed, placed in a plastic quart‐sized bag, and thawed in a water bath at 60°C until they reached room temperature. Berries were manually macerated, the juice was filtered using cheesecloth, and TSS was determined using a handheld refractometer (Master, Atago USA, Inc., Bellevue, WA). Juice pH was determined using a benchtop pH meter (Orion Star A111, Thermo Fisher Scientific, Inc., Waltham, MA, USA), and TA was determined using an autotitrator (G20, Mettler Toledo, Columbus, OH, USA). A 1 ml subsample of juice was used to assess YAN using commercial enzyme assay kits (Vintessential Laboratories, Victoria, AU).
4
pH Measurement of HIU-Treated Protein
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The pH values of the HIU-treated MP solutions adjusted to a protein concentration of 5 mg/mL were measured using a pH meter (Orion Star A111, Thermo Fisher Scientific, Waltham, MA, USA) at room temperature.
5
Raw Milk Chemical and Microbial Analysis
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The analysed chemical parameters of the raw milk samples were pH and titratable acidity (TA). The pH was measured using a digital pH meter (Orion Star™ A111; Thermo Fisher Scientific) and the TA was determined by titration method (39 ) at different time intervals (0, 3, 6 and 9 h). Each sample was measured in triplicate. Total plate count (TPC) and coliform counts were also tested simultaneously by the sample dilution pour plate method. Briefly, tenfold serial dilutions of samples were made up to volume fraction of 10-7 in normal saline solutions (0.85%). A volume of 1 mL of milk sample was diluted in a series of normal saline solutions (9 mL of saline down to 10-8) and 1-mL aliquots of milk in the saline solution from appropriate volume fraction dilutions: 10-1, 10-3 or
10-5 for coliform count, and 10-3, 10-5 or 10-7 for TPC were transferred to Petri dishes. Samples were plated in triplicates, using pour plate technique. Nutrient agar medium (HiMedia) was used for the TPC, while violet red bile agar (HiMedia) was used for counting coliforms incubated at 37 °C. After incubation, plates containing 30 to 300 colonies were selected for screening and results were expressed as log CFU/mL.
6
Evaluating Bacterial Stress Responses
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The isolates were grown in MRS broth at 37 °C for 14 h. An aliquot of 100 µL of each culture was transferred to the experimental MRS. Three experimental MRS were prepared. One set of MRS media was subjected to pH adjustment with 1 M of HCl to 2, 3, and 4 using pH meter (Thermo Scientific, Waltham, MA, USA, Orion Star A111.). A second set of experimental MRS broth was supplemented with sodium chloride (NaCl) to final concentrations of 3%, 6%, 9%, and 12% (w/v, 99.5% purity, M.W. 58.44). The third set of experimental MRS was supplemented with 0.2%, 0.4%, and 0.6% phenol (Sigma-Aldrich, St. Louis, USA, M.W. 94.11). Optical density at 600 nm was measured from each culture after incubation for 24 h. All of those isolates growing in regular MRS broth were regarded as control. Relative growth was determined by the ratio of OD600 of LAB cultures grown in the MRS supplemented with different concentration of acid, NaCl, and phenol to that in the regular MRS.
7
Fruit Extract Physicochemical Analysis
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pH, TSS, and TA were measured in the extract of the fruits (20 g of pulp were macerated and filtered with a fine cloth) in triplicate for each batch during the time interval. The pH was measured using a digital potentiometer (Orion Star A111, Thermo Scientific, USA). TSS was assessed using a digital handheld refractometer (Reichert, Germany), and results were expressed as °Bx. TA content was measured by titrating the samples with NaOH solution at 0.1 M in a pH = 8.1 ± 0.2. The results were expressed as a percentage of citric acid. TSS/TA ratio was calculated as a percentage between the TSS and TA.
8
Pectin-Based Nanocellulose Coatings
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The coatings were prepared as described by Chaichi et al. (2017) (link), with some modifications. Briefly, 6 g of pectin were dissolved in 120 ml of distilled water at 70 °C (Corning, USA). Next, glycerin was added at 0.3 w/w of pectin. The pH value of the solution was adjusted to 4.5 with 0.1 N NaOH (Orion Star A111, Thermo Scientific, USA). After this, different concentrations (0.0, 0.5, 2.0 and 5.0 % w/w) of nanocellulose were added. The dispersions were homogenized in an ultrasonic homogenizer (MicroDisTec™ Homogenizer 1000, Switzerland) at 10.000 rpm for 15 min. Samples were labeled according to the concentration of cellulose nanofibers, as follows: P (0 %), P + CN0.5%, P + CN2%, and P + CN5%.
9
Soil and Compost Characterization Protocol
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The soil bulk density was measured using 100 cm3 stainless-steel soil cores [24 ], and the soil texture was classified using a hydrometer based on the United States Department of Agriculture (USDA) triangle method. Soil pH and electrical conductivity (EC) were measured at a soil:distilled water ratio of 1:5 (w/v), or manure-based compost:distilled water ratio of 1:10 (w/v) using a pH meter (Orion Star™ A111, Thermo Fisher Scientific, Waltham, MA, USA) and an EC meter (SevenCompact Conductivity Meter S230, Mettler Toledo, Columbus, OH, USA), according to the Korean Standard Test Method (KSTM ES 07302). Soil organic matter content (SOM) was analyzed using the Walkley–Black method [25 (link)], and the organic content of compost was measured using a loss-on-ignition method with a furnace (Lindberg/Blue M42.5 LC2 Moldatherm Box Furnace, Thermo Fisher Scientific).
10
Grape Chemical Composition and Rotundone Analysis
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In both years and for each experimental unit, fruit chemical composition data [total soluble solids (TSS), pH, and titratable acidity (TA)] were measured on a randomly selected 100-berry sample selected from the frozen clusters collected at harvest. Frozen berry samples were thawed within a plastic zip-lock bag that was heated in a water bath at 60°C, and berries were hand-crushed for juice analysis. Total soluble solids were measured using a hand-held refractometer (Master, Atago USA, Inc., Bellevue, WA) and juice pH was measured using a benchtop pH-meter (Orion Star A111, Thermo Fisher Scientific, Waltham, MA). Titratable acidity was assessed using an autotitrator (G20, Mettler Toledo, Columbus, OH) on a 10 ml juice sample titrated to an endpoint pH of 8.2 with a 0.1 M NaOH solution. Average berry weight was calculated using a 200-berry sample taken from the frozen harvested clusters.
Berry processing for rotundone extraction and analysis followed the protocol used by Homich et al. (2017) (link). Analysis of rotundone was conducted via solid phase microextraction multidimensional gas chromatography–mass spectrometry (SPME-MDGC-MS) at the Australian Wine Research Institute (AWRI, Glen Osmond, SA) using the equipment and protocols outlined in Geffroy et al. (2014) (link).
Berry processing for rotundone extraction and analysis followed the protocol used by Homich et al. (2017) (link). Analysis of rotundone was conducted via solid phase microextraction multidimensional gas chromatography–mass spectrometry (SPME-MDGC-MS) at the Australian Wine Research Institute (AWRI, Glen Osmond, SA) using the equipment and protocols outlined in Geffroy et al. (2014) (link).
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