Compromising chitinase activity decreases eDNA release and biofilm formation in C. albicans . C. albicans Δcht2, Δcht3, Δcht2/ Δcht3 and parental strain CA14 biofilms were grown in 96 well microtitre plates for 24 h at 37°C in RPMI (A) Following biofilm development, biofilms were carefully washed with PBS and incubated in RPMI for a further 24 h, ± 256 μg/ml DNase. Biomass assessed spectrophotometrically by measuring CV absorbance at 570 nm. Also CV stained biofilms were digitally imaged to show the difference in biofilm formation and effect of DNase treatment. §P < 0.05, §§ p < 0.01. (B) C. albicans Δcht2, Δcht3, Δcht2/ Δcht3 and wild type CA14 (×1000) were grown on Thermanox™ coverslips for 24 h at 37°C. Biofilms were then processed and viewed on a JEOL JSM-6400 scanning electron microscope and images assembled using Photoshop software. Note the reduction in biofilm biomass in Δcht3 and Δcht2/ Δcht3. (C) eDNA release was measured after 4 and 24 h using SYBR green I assay, normalised to each isolates biomass. Each isolate was measured in duplicate, on three separate occasions. *p < 0.05, **p < 0.01. (D) Biofilm growth (hyphal cells indicated by arrow head) and accumulation of eDNA (Indicated by arrow) were visualized under a fluorescence microscope (Motic BA400 Colorview system) at an Ex/Em wavelengths of 350/400 nm for calcofluor white and 540/525 nm for propidium iodide. One representative from each group was digitally photographed.