Sabouraud dextrose broth sdb

Manufactured by Liofilchem

Sourced in Italy

Sabouraud Dextrose Broth (SDB) is a general-purpose microbiological culture medium used for the cultivation of fungi and yeasts. It provides the necessary nutrients and growth conditions for these microorganisms.

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Lab products found in correlation

Sabouraud dextrose agar, by Liofilchem (5 mentions) Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions) Tryptic soy broth tsb, by Liofilchem (2 mentions) Sabouraud dextrose broth (sdb), by Liofilchem (2 mentions) Bhi agar, by Liofilchem (1 mentions) Maxq 4000, by Thermo Fisher Scientific (1 mentions) Tryptic soy broth (tsb), by Liofilchem (1 mentions) Sabouraud dextrose agar, by Merck Group (1 mentions) Sabouraud dextrose broth (sdb), by Merck Group (1 mentions)

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Sabouraud dextrose broth (9 citations)

8 protocols using sabouraud dextrose broth sdb

Cultivation and Biofilm Formation of Drug-Resistant Candida

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In this study, 13 drug-resistant Candida isolates, C. albicans (n = 3), C. glabrata (n = 8), C. krusei (n = 1), and C. guilliermondii (n = 1), belonging to the Biofilm Research Group of the Centre o Biological Engineering, were used (57 (link)). The clinical isolates were subcultured from a frozen stock (Sabouraud dextrose broth [SDB; Liofilchem] medium with 20% [vol/vol] glycerol at −80°C ± 2°C) onto Sabouraud dextrose agar (SDA; Liofilchem) plates and incubated for 24 h. Prior to testing, a preinoculum was prepared in SDB with colonies from the SDA plates for 18 h at 37°C under agitation (120 rpm). Next, the cellular suspensions were centrifuged and washed twice with PBS (5,000 × g for 10 min at 4°C).
For biofilm assays, vaginal Candida isolates cells were cultivated in simulated vaginal fluid (SVF). SVF consisted of 58 mM NaCl (Biochem), 18 mM KOH (AppliChem), 2 mM Ca(OH)₂ (Frilabo), 1.75 mM glycerol (Biochem), 6.7 mM urea (Frilabo), 33 mM glucose (Biochem), and 6.7 g/L yeast nitrogen base (YNB;(Difco)). In addition, natural compounds in the vaginal fluid, such as acetic acid (17 mM; pK_a, 54.76) and lactic acid (22 mM; pK_a, 53.85), were added to maintain the pH at 4.2 as described previously by Sosinska et al., Owen and Katz, and Fernandes et al. (58 (link)– (link)60 (link)).

Fernandes L., Costa R., Silva S., Henriques M., Costa-de-Oliveira S, & Rodrigues M.E. (2023). Effect of Vapor-Phase Oregano Essential Oil on Resistant Candida Species Biofilms: Mechanisms of Action. Microbiology Spectrum, 11(2), e05124-22.

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Preparation and Cultivation of C. tropicalis and P. aeruginosa

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C. tropicalis ATCC 750 and P. aeruginosa DSM 22644 were stored at –80 ± 2 °C in broth medium with 20% (v/v) glycerol. Prior to each assay, C. tropicalis and P. aeruginosa strains were subcultured from the frozen stock preparations onto Sabouraud Dextrose Agar (SDA) and Tryptic Soy Agar (TSA) plates, respectively. SDA and TSA were prepared from Sabouraud Dextrose Broth (SDB; Liofilchem, Roseto degli Abruzzi, Italy) or Tryptic Soy Broth (TSB; Liofilchem, Roseto degli Abruzz, Italy), supplemented with 2% (w/v) agar (Liofilchem, Roseto degli Abruzzi, Italy). The plates were then incubated aerobically at 37 °C for 18–24 h.
Pure liquid cultures (pre-inocula) of C. tropicalis were maintained in SDB, whereas P. aeruginosa was grown overnight in TSB. For planktonic and biofilm assays, 0.22 µm filter-sterilized Roswell Park Memorial Institute (RPMI) 1640 medium (Gibco^® by Life Technologies ^TM, Grand Island, NY, USA) at pH 7.0 was used. Unless otherwise stated, all rinse steps were performed either by using 0.9% (w/v) saline solution (NaCl; J.T. Baker, Deventer, The Netherlands) or ultrapure (UP) sterile water.

Fernandes L., Oliveira A., Henriques M, & Rodrigues M.E. (2020). Honey as a Strategy to Fight Candida tropicalis in Mixed-Biofilms with Pseudomonas aeruginosa. Antibiotics, 9(2), 43.

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Microbial Strain Preparation Procedure

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Enterococcus faecalis (ATCC 29212), stored at −80 °C in brain heart infusion broth (BHI broth, PanReac AppliChem, Darmstadt, Germany) with 20% glycerol, and Candida albicans (ATCC 11225) stored at the same temperature in Sabouraud dextrose broth (SDB, Liofilchem, Roseto degli Abruzzi, Italy) with 20% glycerol were used in this study.
The bacterium was subcultured in brain heart infusion agar (BHI agar, Liofilchem, Roseto degli Abruzzi, Italy) and, prior to the experiments, one isolated colony was inoculated in 10 mL of brain heart infusion broth and grown anaerobically at 37 °C overnight. Candida albicans was subcultured in Sabouraud dextrose agar (SDA, Liofilchem, Roseto degli Abruzzi, Italy) and, before each assay, one isolated colony was inoculated in SDB and grown aerobically at 37 °C overnight with stirring. An aliquot of each culture (300 μL) was transferred into a new fresh liquid medium and grown under the same growth conditions until the stationary growth phase. This culture was then used for the assay.

Correia B.L., Gomes A.T., Noites R., Ferreira J.M, & Duarte A.S. (2022). New and Efficient Bioactive Glass Compositions for Controlling Endodontic Pathogens. Nanomaterials, 12(9), 1577.

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Co-Culture of P. aeruginosa and C. albicans

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P. aeruginosa PAO1 and C. albicans SC5314, two model reference strains with known sequenced whole genome, were used throughout this work. Both strains were stored at– 80 ± 2°C in broth medium with 20% (v/v) glycerol. Prior to each assay, P. aeruginosa and C. albicans strains were subcultured from the frozen stock preparations onto Tryptic Soy agar (TSA) and Sabouraud Dextrose agar (SDA) plates, respectively. TSA and SDA were prepared from Tryptic Soy Broth (TSB; Liofilchem, Italy) or Sabouraud Dextrose Broth (SDB; Liofilchem) supplemented with 1.2% w/v agar (Liofilchem). The plates were then incubated aerobically at 37°C for 18–24 h.
Pure liquid cultures (pre-inocula) of P. aeruginosa were grown overnight in TSB whereas C. albicans was maintained in SDB. For planktonic and biofilm assays, 0.22 μm filter-sterilized RPMI 1640 medium (Gibco® by Life Technologies ^TM, Grand Island, NY, USA) at pH 7.0 was used. Unless otherwise stated, all rinse steps were performed either by using 0.9% (w/v) saline solution (NaCl; J.T. Baker, The Netherlands) or ultrapure (UP) sterile water.

Rodrigues M.E., Lopes S.P., Pereira C.R., Azevedo N.F., Lourenço A., Henriques M, & Pereira M.O. (2017). Polymicrobial Ventilator-Associated Pneumonia: Fighting In Vitro Candida albicans-Pseudomonas aeruginosa Biofilms with Antifungal-Antibacterial Combination Therapy. PLoS ONE, 12(1), e0170433.

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Cryptococcus neoformans Clinical Isolates

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A total eight clinical isolates, namely, Cn46, Cn47, Cn91, Cn96, Cn118, Cn158, Cn169, and Cn173, were used in this work. The clinical isolates of C. neoformans were obtained from the HIV-positive patients at the Yaoundé Central Hospital and identified by serotyping by multiplex PCR in a previous study [21 (link)]. Sabouraud Dextrose agar (SDA, Liofilchem) and Sabouraud Dextrose broth (SDB, Liofilchem) were used for the activation of yeasts and antimicrobial assays, respectively.

Bisso B.N., Makuété A.L., Tsopmene J.U, & Dzoyem J.P. (2023). Biofilm Formation and Phospholipase and Proteinase Production in Cryptococcus neoformans Clinical Isolates and Susceptibility towards Some Bioactive Natural Products. The Scientific World Journal, 2023, 6080489.

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Storage and Cultivation of Candida auris

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Candida auris NCPF 8971 was stored at −80 ± 2 °C in Sabouraud Dextrose Broth (SDB; Liofilchem, Roseto degli Abruzzi, Italy) with 20% (v/v) glycerol. Before each assay, C. auris was subcultured onto Sabouraud Dextrose agar (SDA; Liofilchem) and incubated aerobically at 37 °C for 18–24 h. SDA plates were prepared from SDB supplemented with 2% (w/v) agar (Liofilchem).

Fernandes L., Ribeiro R., Costa R., Henriques M, & Rodrigues M.E. (2022). Essential Oils as a Good Weapon against Drug-Resistant Candida auris. Antibiotics, 11(7), 977.

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Microbial Culture Procedures for Research

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Bacteria and yeast isolates were purchased from ATCC (S. aureus (ATCC 25923), P. aeruginosa (ATCC 89033) and C. albicans (ATCC 90028)) and stored at −80 °C in Luria-Bertani (LB) broth (Liofilchem, Roseto degli Abruzzi, TE, Italy) with 50% glycerol. The experiments were carried out with bacteria cultured in Tryptic Soy Broth (TSB) whereas yeasts in Sabouraud Dextrose Broth (SDB) (Liofilchem) in agitation at 150 rpm (MaxQ 4000, Thermo Scientific, Milan, Italy) or plated on Tryptic Soy Agar (TSA) (Scharlab Italia, Milan, Italy) or Sabouraud Dextrose Agar (SDA) (Liofilchem), respectively, and incubation at 35 °C/37 °C (bacteria) and 25 °C (yeasts).

Fontana R., Marconi P.C., Caputo A, & Gavalyan V.B. (2022). Novel Chitosan-Based Schiff Base Compounds: Chemical Characterization and Antimicrobial Activity. Molecules, 27(9), 2740.

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Candida tropicalis Cultivation and Identification

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Three strains of C. tropicalis were used in this study: one reference strain from the American Type Culture Collection (ATCC 750) and two clinical isolates (U69 and U75) obtained from patients with candiduria admitted to the intensive care unit and belonging to the archive collection of the University Hospital in Maringa ´, Parana ´, Brazil. The yeasts were identified by three methods: the MicroScan rapid yeast identification panel (Dade Behring Inc, CA, USA), the classical biochemical method and molecular identification [15] . The strains were kept frozen at -80 °C in Sabouraud dextrose broth (SDB; Liofilchem, Italy) containing 5 % (v v -1 ) glycerol. For each experiment, strains were subcultured on Sabouraud dextrose agar (SDA; Merck, Darmstadt, Germany) for 48 h at 37 °C. Yeast cells were then inoculated in Sabouraud dextrose broth (SDB; Merck) and incubated for 18 h at 37 °C under agitation in an orbital shaker (120 rev/ min). After incubation, yeast cells were harvested by centrifugation at 80009g for 5 min at 4 °C and washed twice with phosphate buffer solution (PBS; pH 7.5; 0.01 mol l -1 ). The remaining pellets were suspended in artificial urine (AU), and the cellular density adjusted to 1 9 10 7 yeasts mL -1 , using a Neubauer chamber. Artificial urine (pH 5.8) was prepared according to Silva et al. [11] .

Negri M., Silva S., Capoci I.R., Azeredo J, & Henriques M. (2016). Candida tropicalis Biofilms: Biomass, Metabolic Activity and Secreted Aspartyl Proteinase Production. Mycopathologia, 181(3-4).

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