The probiotic strain A. faecalis A12C was isolated from Argyrosomus regius gills at the Instituto de Sanidad Animal y Seguridad Alimentaria of the University of Las Palmas de Gran Canaria. The strain was identified at the Microbiology Department of the Hospital Universitario de Gran Canaria Dr Negrín by means of matrix-assisted laser desorption ionization time-of-flight mass spectrometry (MALDI-TOF MS) system (Vitek®MS, Biomerieux, Madrid, Spain). A. faecalis A12C strain cultures were stored at − 80 °C with 20% glycerol (v/v) addition in Brain Heart Infusion broth (PanReac-AppliChem, Darmstadt, Germany). Fresh cultures were made prior the assays, and the strain was aerobically incubated in Trypticase Soy with 5% sheep blood (Becton Dickinson, Franklin Lakes, New Jersey, USA) medium for 18 h at 37 °C with shaking (120 rpm) each 2 days. Before the challenge, the bacteria were centrifuged at 2500×g for 10 min and washed three times with sterile 0.9% saline solution. The bacteria were resuspended in dinking mineral water (Fonteide®), and bacterial concentration was measured with a spectrometer at 600 nm. Mineral water (Fonteide®) was used to adjust the suspension to 6 × 108 CFU/mL. Finally, the suspension was stored at 4 °C and protected from light.
Trypticase soy
Manufactured by BD
Sourced in United States
Trypticase soy is a powdered medium used for the cultivation of a wide variety of microorganisms. It provides the necessary nutrients and growth factors required for the optimal growth and maintenance of various bacterial species.
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Lab products found in correlation
5 protocols using trypticase soy
1
Isolation and Characterization of Probiotic A. faecalis A12C
2
Evaluating Anti-Pseudomonal Resistance Mechanisms
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To determine the impact on preventing or reverting ceftolozane/tazobactam HR, time–kill analyses were performed on P. aeruginosa isolates with confirmed MHR to ceftolozane/tazobactam. The initial ceftolozane/tazobactam-susceptible and post-PAP ceftolozane/tazobactam-resistant isolates were preferentially selected. Antimicrobial exposures were based on estimated free trough concentrations of ceftolozane/tazobactam (8/2 mg/L) and common non-β-lactam antimicrobials used in CF exacerbations, amikacin (2 mg/L) and ciprofloxacin (0.5 mg/L) were tested.21 (link) Each isolate was subcultured twice on trypticase soy agar with 5% sheep blood (Becton, Dickinson & Co., Sparks, MD, USA). MH broth was inoculated with the bacterial suspension of ∼1 × 108 cfu/mL in 5 mL, colourless culture tubes (Falcon, Fisher Scientific). Control experiments without active compound were included. Singular studies were conducted unless initial bacterial log growth across arms was inconsistent. Final volumes for each P. aeruginosa–drug concentration were 5 mL and incubated at 37°C using a shaker at 250 rpm. Samples were taken from each culture tube at 0, 3, 6 and 24 h from the time of broth inoculation. Multiple 1:10 dilutions were made in saline and subcultured onto blood agar plates. Plates were incubated for 18–24 h at 37 °C and mean bacterial densities in cfu/mL were determined for each isolate.
3
Culturing H. pylori for Gastric Cell Co-culture
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Wild-type CagA positive H. pylori 7.13, J166 and the rodent-adapted PMSS1 strain were a kind gift from Dr. Richard Peek, Jr. (Vanderbilt University Medical Center). Briefly, H. pylori strains were cultured in trypticase soy agar with 5% sheep blood (BD Biosciences, Bedford, Massachusetts, USA) for 3 days and transferred to a new agar plate [25 ]. The strains were then cultured in Brucella broth with 10% serum and 10 μg/mL vancomycin at 37 °C and placed in an incubator with 5% CO2 overnight. The H. pylori bacteria was added over gastric cells, for a co-culture, at a multiplicity of infection of 100:1 and cells were harvested at different time points.
4
Bacterial Culture Conditions for Diverse Microbes
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H. pylori strains 26695, G27, 98-10, and isogenic 26695 mutants (19 (link), 20 (link)) were grown on Trypticase soy agar supplemented with 5% sheep blood (BD Biosciences) or brucella broth supplemented with 5% fetal bovine serum (FBS) at 37°C in 5% atmosphere CO2. E. coli MG1655 harboring the conjugative plasmid pKM101, E. coli MS411 harboring the R1-16 conjugative plasmid, and E. coli WM1652 recipient cells were grown on lysogeny broth (LB) agar or broth supplemented with ampicillin (50 µg/ml), kanamycin (50 µg/ml), or tetracycline (20 µg/ml), respectively. Agrobacterium tumefaciens C58 and its derivative GV3101 were maintained on LB plates supplemented with rifampin (25 µg/ml) at 28°C. A. tumefaciens GV3101 harboring pCAMBIA vectors were maintained on LB containing rifampin (25 µg/ml) and kanamycin (100 µg/ml).
5
Isolation and Characterization of a Novel Marine Bacterium
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Bacterial strains and culture conditions A marine dune was collected from Boryeong (36°20'95"N, 126°53'80"E) close to the Yellow Sea of Republic of Korea. The sample (about 1-2 g) was serially diluted with 0.85% (w/v) saline solution and spread onto marine agar 2216 (MA; BD Difco). After incubation at 25 °C for 7 days, strain BSSL-BM10 T was isolated from the MA plate and speak onto trypticase soy agar (TSA; BD Bacto) at 30 °C. Cells of strain BSSL-BM10 T were suspended in a sterile solution containing 20% (w/v) glycerol and stored at -80 °C for long-term preservation. Devosia naphthalenivorans JCM 32509 T and Devosia ribo avina DSM 7230 T were obtained from the Japan Collection for Microorganisms (JCM; Japan) and Deutsche Sammlung von Mikroorganismen und Zellkulturen (DSMZ; Germany), respectively. Cells of strain BSSL-BM10 T and D. ribo avina DSM 7230 T obtained from culture grown for 3 days in trypticase soy broth (BD Bacto) at 30 °C were used to extract DNA and to analyze isoprenoid quinones and polar lipids. Cell masses for cellular fatty acid analysis were obtained under the following conditions: strain BSSL-BM10 T were harvested from TSA plates after cultivation for 3, 5, and 7 days at 30 ºC, and D. naphthalenivorans JCM 32509 T and D. ribo avina DSM 7230 T were harvested from MA and TSA plates, respectively, after cultivation for 5 days at 30 ºC.
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