Culture media profoundly affect Candida albicans and Candida tropicalis growth, adhesion and biofilm development

Abstract

As there are sparse data on the impact of growth media on the phenomenon of biofilm development for Candida we evaluated the efficacy of three culture media on growth, adhesion and biofilm formation of two pathogenic yeasts, Candida albicans and Candida tropicalis. The planktonic phase yeast growth, either as monocultures or mixed cultures, in sabouraud dextrose broth (SDB), yeast nitrogen base (YNB), and RPMI 1640 was compared, and adhesion as well as biofilm formation were monitored using MTT and crystal violet (CV) assays and scanning electron microscopy. Planktonic cells of C. albicans, C. tropicalis and their 1:1 co-culture showed maximal growth in SDB. C. albicans/C. tropicalis adhesion was significantly facilitated in RPMI 1640 although the YNB elicited the maximum growth for C. tropicalis. Similarly, the biofilm growth was uniformly higher for both species in RPMI 1640, and C. tropicalis was the slower biofilm former in all three media. Scanning electron microscopy images tended to confirm the results of MTT and CV assay. Taken together, our data indicate that researchers should pay heed to the choice of laboratory culture media when comparing relative planktonic/biofilm growth of Candida. There is also a need for standardisation of biofilm development media so as to facilitate cross comparisons between laboratories.

Fluconazole impacts the extracellular matrix of fluconazole-susceptible and -resistant Candida albicans and Candida glabrata biofilms

“…The microorganisms kept at −80°C were seeded onto Petri dishes with Sabourand dextrose agar (DIFCO, Detroit, MI) culture medium supplemented with chloramphenicol (50 mg/L) [28] and incubated at 37°C for 48 h. Then, five colonies of each microorganism were added separately to tubes containing Yeast Nitrogen Base medium (YNB – DIFCO, Detroit, MI) supplemented with 100 mM of glucose [28] and these pre-inocula were incubated at 37°C for 16 h. Subsequently, the pre-inocula were diluted with fresh YNB medium plus 100 mM glucose (1:20 dilution for C. albicans strains and 1:10 for C. glabrata strains) to form the inocula, and the OD 540 nm and CFU were determined every 2 h in a total of 16 h for each strain. Biofilms were generated using mid-log growth phase cells (Figure 1).…”

Influence of Laboratory Culture Media on in vitro Growth, Adhesion, and Biofilm Formation of <b><i>Pseudomonas aeruginosa</i></b> and <b><i>Staphylococcus aureus</i></b>

“…Some even fail to develop biofilm complexes of bacteria found in differing niduses depending on their role within the biofilm [10]. Additionally, with the growing attention on biofilm infections and the need for finding novel therapeutic strategies to combat these infections, it is essential to standardize relevant techniques across laboratories.…”

The Effect of Nutritive and Non-Nutritive Sweeteners on the Growth, Adhesion, and Biofilm Formation of Candida albicans and Candida tropicalis

“…A 100-µL sample of prepared Candida cell suspensions was inoculated in triplicate into a 96-well, flat-bottom, microtiter plate. The growth rates were then determined by optical density (OD 492 ) measurements in each well at 492 nm at 2-h intervals for 14 h using a microtiter plate reader (SpectraMax Plus 384; Molecular Devices, Inc., USA) and growth curves were prepared [8,11].…”

“…To examine the initial adhesion phase, 100 µL of the prepared standard cell suspensions were inoculated in triplicate wells of a microtiter plate and incubated for 90 min at 37 ° C [6,8]. After 90 min of incubation, the plates were carefully washed twice with 200 µL sterile PBS to remove non-adherent cells, and the adherent cells were quantified using a tetrazolium salt 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay (Sigma-Aldrich) [20].…”

“…Both C. albicans and C. tropicalis are known to produce extensive oral biofilms both in vitro and in vivo, and some are macroscopically visible as white, pseudo-membranes in disease such as oral thrush [8][9][10]. Furthermore, it is known that fermentable dietary sugars including sucrose and glucose promote Candida growth, adhesion, and biofilm development [11] and could aggravate oral diseases, in particular Candida-associated denture stomatitis and dental caries [12].…”