Tryptone soya broth (tsb)

Manufactured by Thermo Fisher Scientific

Sourced in United Kingdom, United States, Italy, Germany, Australia

Tryptone Soya Broth (TSB) is a general-purpose culture medium used for the growth and cultivation of a wide range of microorganisms, including bacteria, yeasts, and fungi. It provides a nutritious environment that supports the growth and proliferation of these microorganisms.

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165 protocols using tryptone soya broth (tsb)

Bacterial and Fungal Propagation Protocols

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The bacterial isolates were propagated after Soliman et al. (2021b , 2021c ). Escherichia coli O157: H7 suspension (2.0 × 10⁴ CFU.ml^-1) was propagated into Mac-Conkey broth (Oxoid™) at 44°C/24 hours, eosin methylene blue agar (Oxoid^TM) at 37°C/24 hours, and tryptone soya broth (Oxoid™) providing 1.7 × 10¹¹ CFU. ml^-1 suspension. Lyophilized S. Typhimurium (3.4 × 10² CFU) was propagated into Rappaport-Vassiliadis broth (Oxoid™) at 37°C/24 hours, xylose lysine deoxycholate agar (Oxoid™) at 37°C/24 hours, and tryptone soya broth providing 1.5 × 10⁶ CFU.ml^-1 suspension.
The fungal isolates were propagated following Soliman et al. (2021b) . Aspergillus niger and C. albicans clinical isolates were propagated into sabouraud dextrose broth (SDB; HIMEDIA^®) at 37°C/24–72 hours, sabouraud dextrose agar (SDA; Oxoid™) at 37°C/24 hours, identified with lactophenol cotton blue stain (Hardy Diagnostics^®), and resuspended in SDB broth, providing 2.5 × 10⁸ CFU.ml^-1suspensions.

Soliman E.S., Hassan R.A, & Farid D.S. (2023). Eichhornia crassipes from wastes to valuable products in water purification and influence on growth and impregnability in overwhelmed broiler chickens. Open Veterinary Journal, 13(4), 451-465.

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Isolation and Characterization of K. pneumoniae Phage

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K. pneumoniae strains isolated from the caecum were grown to OD₆₆₀ ∼ 0.4 in tryptone soya broth (Oxoid Ltd), and used in spot assays as follows. An aliquot of culture (200 µl) was inoculated into 3 ml tryptone soya broth containing 0.3% (w/v) agarose (SeaKem LE agarose; Lonza Rockland) that had been heat-treated by microwaving and dispensed aseptically from a larger volume before cooling to 48 °C. The overlays containing bacteria were poured over 20 ml solid agar plates of autoclaved tryptone soya agar (Oxoid Ltd). Once the agar had solidified, a 10 µl spot of filtered caecal effluent was applied to the overlays, and the plates were incubated overnight at 37 °C in the anaerobic cabinet. Identical plaques were observed on all K. pneumoniae isolates; strain L4-FAA5 was used to propagate and purify an isolated phage in tryptone soya broth or reinforced clostridial and nutrient media (Oxoid Ltd). Neither calcium nor magnesium was added to media during propagations.
The ability of the purified phage to infect a panel of K. pneumoniae subsp. pneumoniae clinical isolates (Table 1) was determined using the spot assay as described above, except that nutrient agar plates were used for the base. Strain L4-FAA5 was used as a positive control.

Hoyles L., Murphy J., Neve H., Heller K.J., Turton J.F., Mahony J., Sanderson J.D., Hudspith B., Gibson G.R., McCartney A.L, & van Sinderen D. (2015). Klebsiella pneumoniae subsp. pneumoniae–bacteriophage combination from the caecal effluent of a healthy woman. PeerJ, 3, e1061.

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Blood and Stool Specimen Collection

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After the interview, venous blood and stool specimens were collected aseptically from each study participant. Ten ml and 3 ml blood specimens were collected aseptically from adults and children respectively using culture bottles with tryptone soya broth (Oxoid; Hampshire UK). Besides, fresh stool specimens were collected using sterile screw capped containers. Both specimens were transported to medical microbiology research laboratory of Bahir Dar University College of Medicine and Health Sciences within two hours of collection.

Amsalu T., Genet C, & Adem Siraj Y. (2021). Salmonella Typhi and Salmonella Paratyphi prevalence, antimicrobial susceptibility profile and factors associated with enteric fever infection in Bahir Dar, Ethiopia. Scientific Reports, 11, 7359.

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Ciprofloxacin-loaded PLGA/Chitosan Nanoparticles for Dental Applications

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Ciprofloxacin hydrochloride was supplied by EPICO Company, Cairo, Egypt. Poly (d,l lactide-co-glycolide) ‘PLGA’ lactide: glycolide 50:50 ester terminated molecular weight 38,000–54,000 Da, chitosan low molecular weight 50,000–190,000 Da and Poly vinyl alcohol molecular weight 31,000–50,000 were supplied by Sigma Aldrich, Berlin, Germany. Brain heart infusion broth was supplied by LAB M United Kingdom, Crystal Violet supplied by Oxford Lab Chem, Mumbai, India, Tryptone soya broth was supplied by OXOID, England. Cellophane membrane 12–14 kDa supplied by Spectrum Medical Inc, Raleigh, NC. Normal saline for IV injection and Glucose 5% for IV injection supplied by Nasr Company, Cairo, Egypt. Ethanol was supplied by Fisher Scientific, Loughborough, UK. Hydrochloric acid was supplied by Al Ahram Laboratory Chemicals CO, Cairo, Egypt Dichloromethane and Acetic acid were supplied by Pio Chem, Cairo, Egypt. Human extracted single rooted teeth provided from dentistry hospital of the British University in Egypt.

Arafa M.G., Mousa H.A, & Afifi N.N. (2019). Preparation of PLGA-chitosan based nanocarriers for enhancing antibacterial effect of ciprofloxacin in root canal infection. Drug Delivery, 27(1), 26-39.

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Crystallization Assay of Urinary Solutes

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Ethylenediaminetetraacetic acid tetrasodium salt hydrate (≥99.0%) (tEDTA), calcium chloride dihydrate, magnesium chloride hexahydrate (≥99.0%), sodium chloride (≥99.0%), sodium sulfate (≥99.0%), trisodium citrate dihydrate (≥99.0%), potassium chloride (≥99.0%), ammonium chloride (≥99.5%) and urea were obtained from Sigma-Aldrich (Dorset, UK). Creatinine (98%), sodium oxalate (≥99.5%) and potassium dihydrogen phosphate (≥98.0%) were purchased from Alfa Aesar (Heysham, UK). Quarter-strength Ringer's solution (QSRS), phosphate-buffered saline (PBS), tryptone soya broth (TSB), Mueller-Hinton broth (MHB) and agar were purchased from Oxoid Ltd (Hampshire, UK). BARD silicone, 14-channel male catheters and URIPLAN 2 L drainable bed bags (98 cm inlet) were purchased from BARD Ltd (Crawley, UK). Proteus mirabilis ATCC 51286 and Staphylococcus aureus ATCC 29213 were purchased from LGC Standards (Middlesex, UK), and Escherichia coli NSM59 was obtained from Dr Jones (University of Bath, UK). Deionised water used in bulk crystallisation assays was purified with an Aqua Solutions RODI-C-12A purification system (18.2 MΩ).

Moore J.V., Kim D., Irwin N.J., Rimer J.D, & McCoy C.P. (2023). Tetrasodium EDTA for the prevention of urinary catheter infections and blockages. RSC Advances, 13(4), 2202-2212.

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Culturing SpPdp11 Strain for Beneficial Aquaculture

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The SpPdp11 strain was grown in tryptone soya broth (OXOID Ltd., Basingstoke, UK) supplemented with 1.5% NaCl (TSBs, 20 h, 20 °C) under continuous shaking. Contant shaking was used to wash the bacteria that were removed from the plates in sterile phosphate-buffered saline (PBS, pH 7.4). The density of the bacterial suspension was resolved utilizing Coulter Z2 particle counter (BECKMAN Coulter, Barcelona, Spain), and the volume was adjusted to the target concentration (10⁹ cfu g⁻¹). This dose was chosen based on the health benefits in S. aurata and S. senegalensis as reported in previous studies [27 (link),29 (link)]

Cerezo I.M., Pérez-Gómez O., Bautista R., Seoane P., Esteban M.Á., Balebona M.C., Moriñigo M.A, & Tapia-Paniagua S.T. (2023). SpPdp11 Administration in Diet Modified the Transcriptomic Response and Its Microbiota Associated in Mechanically Induced Wound Sparus aurata Skin. Animals : an Open Access Journal from MDPI, 13(2), 193.

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Endophyte and Vibrio Cholerae Antibiofilm Protocol

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The production was conducted using a liquid medium in Erlenmeyer flasks for endophyte bacteria and Vibrio cholerae strains in. One loop of bacteria was inoculated into 20 mL of brain heart infusion broth (Oxoid) as a production medium and incubated at 37 °C with 120 rpm agitation for 24 h. While for actinomycetes, the isolate was inoculated into 20 mL tryptone soya broth (Oxoid) as a production medium and incubated at 28 °C with agitation at 120 rpm for 7 days as instructed in the previous study [26 ]. The culture was transferred into centrifuge tubes and centrifuged at 7798 RCF (Thermo Scientific) for 10 min. The supernatant was collected and transferred to centrifuge tubes, and subsequently centrifuged again at 7798 RCF (Thermo Scientific) for 10 min. After the second centrifugation, the supernatant was collected and used for the antibiofilm assays or stored at 4 °C for a week or at -20 °C for a month.

M.i.c.h.a.e.l, & Waturangi D.E. (2023). Antibiofilm activity from endophyte bacteria, Vibrio cholerae strains, and actinomycetes isolates in liquid and solid culture. BMC Microbiology, 23, 83.

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Stimulating Rainbow Trout Splenocytes

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The Gram-negative fish pathogen A. salmonicida subsp. salmonicida CECT4237 was aerobically grown in Tryptone Soya Broth (Oxoid) at 25 ˚C. To stimulate the rainbow trout splenocytes, A. salmonicida grown in broth overnight to exponential phase was heat-inactivated at 65 °C for 1 h. In some experiments, LPS obtained from A. salmonicida was used to stimulate the cells. For this, LPS was isolated using a commercial extraction kit (iNtRON Biotechnology) following the manufacturer’s protocol. The absence of DNA and protein contamination was confirmed by SDS-PAGE and agarose electrophoresis.

Soleto I., Morel E., Muñoz-Atienza E., Díaz-Rosales P, & Tafalla C. (2020). Aeromonas salmonicida activates rainbow trout IgM+ B cells signalling through Toll like receptors. Scientific Reports, 10, 16810.

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Cultivation of Diverse Microorganisms

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B. pumilus (formerly known as Bacillus sp. strain 13M) [7 (link)], C. boidinii (strain 682) [8 (link)] and T. harzianum, strain Var119 [9 (link),10 (link)], were used throughout this study. The bacterium was maintained on Tryptone Soya Agar (Oxoid, Basingstoke, UK) at 4 °C and grown in Tryptone Soya broth at 30 °C for 48 h; the yeast was maintained on YPG agar (yeast extract, 10 g/L; peptone, 20 g/L; glucose, 20 g/L) and grown in YPG broth incubated at 25 °C for 72 h, while the fungus was maintained on PDA (potato, 200 g/L; dextrose, 20 g/L; agar 20 g/L) at 4 ± 3 °C, and grown on PDA at 23 ± 2 °C for 15 days in the dark.

Campaniello D., Carlucci A., Speranza B., Raimondo M.L., Cibelli F., Rosaria Corbo M, & Bevilacqua A. (2020). A Comparative Study on Trichoderma harzianum and a Combination of Candida/Bacillus as Tools for the Bioremediation of Table Olive Processing Water. Microorganisms, 8(6), 878.

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Bacterial Strain Cultivation and Preparation

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The bacterial strains used in this study were: Enterococcus faecium LMG 11423, Staphylococcus aureus NCTC 4135, Klebsiella pneumoniae LMG 3081, Acinetobacter baumannii LMG 1041, Pseudomonas aeruginosa LMG 9009 and Enterobacter cloacae LMG 2783 (LMG: Laboratorium voor Microbiologie, Universiteit Gent, Belgium; NCTC: National Collection of Type Cultures, Colindale, UK). Bacteria were cultured in 100 mL nutrient broth (Oxoid Ltd, UK), with the exception of E. faecium and A. baumannii which were cultured in tryptone soya broth (Oxoid Ltd, UK), at 37 °C under rotary conditions (120 rpm) for 18–24 h (C24 Incubator Shaker, New Brunswick Scientific, USA). Following cultivation, broths were centrifuged at 3939 × g for 10 min (Heraeus Labofuge 400R; Kendro Laboratory Products, UK), and the cell pellet was resuspended and serially diluted in phosphate buffered saline (PBS; Oxoid Ltd, UK) to obtain the required population density, in colony forming units per millimetre (CFUmL⁻¹), for experimental use. Population densities were checked by cultivation on nutrient agar plates (NA; Oxoid Ltd, UK), with the exception of A. baumannii and E. faecium, which were cultivated on tryptone soya agar (TSA; Oxoid Ltd, UK).

Sinclair L.G., Anderson J.G., MacGregor S.J, & Maclean M. (2024). Enhanced antimicrobial efficacy and energy efficiency of low irradiance 405-nm light for bacterial decontamination. Archives of Microbiology, 206(6), 276.

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