Sabouraud dextrose broth (sdb)

Manufactured by Merck Group

Sourced in Germany, United States, France, Italy

Sabouraud dextrose broth is a microbiological culture medium used for the isolation and cultivation of fungi. It provides a nutrient-rich environment that supports the growth of a wide range of fungal species. The broth contains dextrose as the primary carbon source and peptones to supply essential nutrients for fungal metabolism and proliferation.

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Dextrose (252 citations) Sabouraud broth (20 citations) Sabouraud dextrose (8 citations) Sabouraud dextrose broth (SDB (5 citations) Sabouraud dextrose broth medium (SDB; (2 citations)

91 protocols using sabouraud dextrose broth (sdb)

Microdilution Assay for Antifungal Evaluation

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The microdilution method was performed according to the method described by Marcello et al. [34 ] and Barbour et
al. [35 (link)], with certain modifications described in our previous research [33 ]. The following
media were used in this method: Sabouraud Dextrose Broth (SDB; Merck, Darmstadt) and SDA. Inoculated microtiter plates were incubated at 25°C for 48 hr, and inoculated SDA plates
were incubated at 25°C for 72 hr. The lowest LA concentration at which there was no visible fungal growth in SDB was considered to be the minimum inhibitory concentration (MIC).
The lowest LA concentration at which there was no growth on the SDA after re-inoculation from the liquid medium (SDB) was considered to be the minimum fungicidal concentration
(MFC).

STANOJEVIĆ-NIKOLIĆ S., DIMIĆ G., MOJOVIĆ L., PEJIN J., RADOSAVLJEVIĆ M., ĐUKIĆ-VUKOVIĆ A., MLADENOVIĆ D, & KOCIĆ-TANACKOV S. (2020). Reduction of sterigmatocystin biosynthesis and growth of food-borne fungi by lactic acid. Bioscience of Microbiota, Food and Health, 39(3), 83-88.

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Bimetallic NPs Modulate C. albicans Morphogenesis

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The effect of bimetallic NPs on C. albicans SC5314 morphogenesis was determined according to the previously reported method [38 (link)]. Briefly, the yeast cells from agar slants were grown in SDB (SigmaAldrich, St. Louis, MO, USA) at 37 °C until the late log phase was achieved. Subsequently, with the help of phosphate-buffered saline (PBS), cells were washed twice and transferred into fresh SDB, and re-incubated further for 48 h at 37 °C resulting in a synchronized yeast population. For initiation of yeast to hyphae transition, the synchronized yeast cells (50 μL) were added to a fresh SDB (10 mL) medium with 10% fetal bovine serum (SigmaAldrich, St. Louis, MO, USA). To this mixture, bimetallic NPs at desired concentrations (¼ × MIC and ½ × MIC) were added and incubated at 37 °C. The external pH of the medium was kept constant at pH 6.5. The cell aliquots were collected at different time intervals, and the morphological changes were visualized using a microscope (Leica Microsystems, Heerbrugg, Switzerland). Untreated yeast cells were also included in the study as a control.

Kamli M.R., Malik M.A., Lone S.A., Sabir J.S., Mattar E.H, & Ahmad A. (2021). Beta vulgaris Assisted Fabrication of Novel Ag-Cu Bimetallic Nanoparticles for Growth Inhibition and Virulence in Candida albicans. Pharmaceutics, 13(11), 1957.

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Standardized Candida glabrata Culture Protocol

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For each experiment, C. glabrata ATCC2001, C. glabrata 534784, and C. glabrata 562123 strains were subcultured on Sabouraud dextrose agar (SDA) (Merck, Darmstadt, Germany) for 24 h at 37 °C. The cells were then inoculated in Sabouraud dextrose broth (SDB) (Merck) and incubated for 18 h at 37 °C under agitation at 120 rpm. After incubation, the cells were harvested by centrifugation at 3000× g (Thermo Scientific, CL10, Hampton, NH, USA) for 10 min at 4 °C and washed twice with phosphate buffered saline (PBS, pH = 7.5). The cell pellets were then suspended in Roswell Park memorial institute (RPMI), and the cellular density was adjusted to 1 × 10⁵ cells/mL, using a Neubauer counting chamber.

Rodrigues C.F, & Henriques M. (2018). Portrait of Matrix Gene Expression in Candida glabrata Biofilms with Stress Induced by Different Drugs. Genes, 9(4), 205.

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Comparative Evaluation of Fluconazole-Susceptible and Resistant Candida albicans Strains

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In the present study, a FLC-susceptible C. albicans ATCC 10231 and a clinical FLC-resistant C. albicans PFCC 93-902 were obtained from the Pathogenic Fungi Culture
Collection of the Pasteur Institute of Iran, Tehran, Iran, were examined. C. albicans strains were kept as a frozen stock in glycerol at 80 ºC. Throughout the
investigation, fresh fungal cultures were generated by sub-culturing on Sabouraud dextrose agar (SDA, Merck, Germany) at 35°C for 24 h. To make the cell suspension,
one colony from the SDA cultures was taken and re-suspended in Sabouraud dextrose broth (SDB, Merck, Germany) at a concentration of 1×10⁶ cells/ml [ 2 (link) ].

Nademi A., Shams-Ghahfarokhi M, & Razzaghi-Abyaneh M. (2021). Effect of Allium cepa loaded polyacrylonitrile and polyvinyl pyrrolidone nanofibers on Candida albicans growth and the expression of CDR1 and CDR2 genes. Current Medical Mycology, 7(4), 28-33.

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Standardized Candida glabrata Culture

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The reference strain from the American Type Culture Collection, C. glabrata ATCC 2001, was used in this study. For each experiment, C. glabrata ATCC 2001 was subcultured on Sabouraud dextrose agar (SDA) (Merck, Darmstadt, Germany), for 24 h at 37 °C. Cells were then inoculated in Sabouraud dextrose broth (SDB) (Merck, Darmstadt, Germany), and incubated for 18 h at 37 °C under agitation at 120 rpm. After incubation, the cells were harvested by centrifugation at 3000 g for 10 min at 4 °C, and washed twice with Phosphate Buffered Saline (PBS, pH = 7.5). Pellets were then suspended in RPMI-1640 (Sigma-Aldrich, St. Louis, MI, USA), and the cellular density was adjusted to 1 × 10⁵ cells/mL, using a Neubauer counting chamber [28 (link)].

Rodrigues C.F., Alves D.F, & Henriques M. (2018). Combination of Posaconazole and Amphotericin B in the Treatment of Candida glabrata Biofilms. Microorganisms, 6(4), 123.

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Quantifying Candida Biofilm Formation

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In the present study, biofilm formation was determined using preseterilized polystyrene 96-well microplates (SPL, Korea) (20 ). For each isolate, a suspension from an overnight culture on SDA was prepared in sterile distilled water and adjusted to 1 McFarland. Each well of the microplate was filled with 180 μL of Sabouraud dextrose broth (SDB) (Merck, Germany) supplement with 8% glucose and then 20 μL of the standard suspension of tested isolates was inoculated (21 (link)). Microplates were covered with lids and incubated at 35°C for 24 hours (20 ). The medium in wells was removed and washed three times with sterile phosphated buffer solution (PBS). Microplates were stained with Giemsa for 5 minutes and then read at 405 nm by an Elisa reader (21 (link), 22 (link)). All tests were done in triplicates and means were calculated. Finally, adherent biofilm layers were scored as either negative; weak (+) (percentage transmittance (%T ≤ 20)); moderate, (++) (%T = 20-35); strong (+++) (%T = 36-50) and very strong (++++) (%T ≥ 50) (21 (link)).

Zarei Mahmoudabadi A., Zarrin M, & Kiasat N. (2014). Biofilm Formation and Susceptibility to Amphotericin B and Fluconazole in Candida albicans. Jundishapur Journal of Microbiology, 7(7), e17105.

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Synthesis and Characterization of Zinc Nanoparticles

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Zinc nitrate hexahydrate (Zn(NO₃)₂·6H₂O; 98% purity; CAS number 10196-18-6), potassium hydroxide (KOH; 90% purity; CAS number 1310-58-3), glutaraldehyde solution (OHC(CH₂)₃CHO; 50% in H₂O; CAS number 111-30-8), tween-80 (CAS number 9005-65-6), methanol (CH₃OH, CAS number 67-56-1), methanol HPLC grade (CH₃OH, CAS number 67-56-1), sodium chloride (NaCl, CAS number 7647-14-5), and Sabouraud dextrose broth (SDB) were obtained from Merck KGaA (Darmstadt, Germany). Dehydrated culture media: potato dextrose agar (PDA) was procured from BD Bioxon (Becton Dickinson de Mexico SA de CV, Estado de Mexico, Mexico). Bromine solution (0.03%) was purchased from Vicam (Milford, MA, USA). All solutions were prepared using deionized water.

Hernández-Meléndez D., Salas-Téllez E., Zavala-Franco A., Téllez G., Méndez-Albores A, & Vázquez-Durán A. (2018). Inhibitory Effect of Flower-Shaped Zinc Oxide Nanostructures on the Growth and Aflatoxin Production of a Highly Toxigenic Strain of Aspergillus flavus Link. Materials, 11(8), 1265.

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Wheat Sourdough Microbial Community

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Refined flour (Sinangil, Eksim Co., İstanbul, Türkiye), whole wheat flour (Sinangil, Eksim Co., İstanbul, Türkiye), dry yeast (Pakmaya, İzmit, Türkiye), and salt (Billur, İzmir, Türkiye) were purchased from a local market. MRS (de Man, Rogosa, and Sharpe) agar, SDA (Sabouraud dextrose agar), MRS broth, Sabouraud dextrose broth (SDB), and peptone water were purchased from Merck (Darmstadt, Germany). Lactic acid bacteria (Lacp. plantarum LABE 29, Levilactobacillus brevis LABE 32) and Saccharomyces cerevisiae TGM 55 were obtained from the culture collection of Yildiz Technical University in the Department of Food Engineering. The isolation source of these bacteria and yeasts was wheat sourdough.

, & Ozulku G. (2024). Monitoring the Dough Properties, Quality Characteristics and Volatile Compounds of Whole Wheat Bread Made by Different Sourdough Types during Frozen Storage. Foods, 13(9), 1388.

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Candida glabrata Strain Cultivation

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For each experiment, C. glabrata ATCC2001 and C. glabrata ΔHT6 strains were subcultured on Sabouraud dextrose agar (SDA) (Merck, Darmstadt, Germany) and the mutant C. glabrata Δmnn2 was cultured on SD-his, as indicated by West et al. [9 (link)], during 24 h at 37 °C. Cells were then inoculated in Sabouraud dextrose broth (SDB, Merck) and incubated for 18 h at 37 °C under agitation at 120 rpm. After incubation, the cells were harvested by centrifugation at 3000 g for 10 min at 4 °C and washed twice with phosphate buffered saline (PBS 0.1M, pH = 7.5). Pellets were then suspended in RPMI-1640 (Sigma-Aldrich, Roswell Park, St. Louis, MO, USA) and the cellular density was adjusted to 1 × 10⁵ cells/mL, using a Neubauer counting chamber.

Rodrigues C.F., Vilas Boas D., Haynes K, & Henriques M. (2018). The MNN2 Gene Knockout Modulates the Antifungal Resistance of Biofilms of Candida glabrata. Biomolecules, 8(4), 130.

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Cultivation and Characterization of Candida albicans

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Candida albicans SC5314 strain was used in this study. This strain belongs to the Biofilm group collection, located at the Centre of Biological Engineering of Minho University (Braga, Portugal), and its identity was confirmed by PCR-based sequencing with specific primers (ITS1 and ITS4) [20 (link)]. For all experiments, cells were subcultured on sabouraud dextrose agar (SDA; Merck, Germany) and incubated for 24 h at 37 °C. An inoculum, obtained from SDA plates, was resuspended in sabouraud dextrose broth (SDB; Merck, Germany) and incubated at 37 °C for 18 h at 120 rpm. After this time, the cells’ suspension was centrifuged for 10 min at 3000 g and 4 °C and washed twice with phosphate-buffered saline (PBS; pH 7, 0.1 M). Pellets were resuspended in 5 mL of PBS and the cellular density adjusted for each experiment using a Neubauer chamber (Marienfild, Land-Konicshofem, Germany) to 1 × 10⁶ cells ml⁻¹.

Barbosa A., Araújo D., Ribeiro E., Henriques M, & Silva S. (2020). Candida albicans Adaptation on Simulated Human Body Fluids under Different pH. Microorganisms, 8(4), 511.

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