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Prevention of Biofilm Formation
In this experiment, to determine whether jujube honey
could prevent the formation of Candida biofilms and to
determine the lowest concentration of honey capable of
preventing biofilm formation, different concentrations of
honey in YEPD broth (80% w/v, 40% w/v, 20% w/v,
10% w/v and 5% w/v) were used to study the kinetics of
biofilm inhibition. The inhibition of biofilm formation
was dependent on the concentration of the honey. It was
evident that concentrations of honey below 10% w/v did
not inhibit the biofilm and even encouraged biofilm development
(Figure 2). However, concentrations more than
10% w/v inhibited significantly the biofilm formation.
Effect of Honey on Established Biofilms Similarly, when 24 h established biofilms were treated with
different concentrations of jujube honey (80e5% w/v), the
C. albicans biomass was significantly reduced after 24 h of
contact with honey concentrations greater than 10% w/v,
but biofilm growth was enhanced at 5% w/v. A higher concentration
of jujube honey was required to disrupt established
biofilms than to prevent biofilm formation.
When a 24-h established biofilm was
treated with 40% w/v of honey, growth of the established
biofilm was inhibited, and some small pores developed in
the cell walls. These pores may be due to bursting of cell
membrane of C. albicans cells by shrinkage and osmotic effect
of honey, which led to cell death and to a reduction in
the numbers of established cell (Figure 4B). No exopolysaccharide
material is observed and shrinkage of cell membrane
due to plasmolysis has been observed (Figure 4B).
Biofilm formed in the presence of 40% (w/v) jujube honey,
no exopolysaccharide material and cell aggregation are
observed. Shrinkage of the cell membrane indicates cell
lysis (Figure 4C and 4D).
differences between the untreated and treated C. albicans
biofilms in this region indicated that honey affected the
formation and secretion of exopolysaccharide matrix by
altering the sugars (major constituents of C. albicans biofilm
exopolysaccharides) composition and deposition.
Thus, there is direct evidence that honey affects the exopolysaccharide
composition of C. albicans biofilms.
Mature C. albicans biofilms are very difficult to eradicate
and are recalcitrant to antifungals. The extracellular
glucan present in extracellular matrix is required for C. albicans
biofilm resistance and it acts by sequestering antifungals,
rendering cells resistant to their action (51).
Many antimicrobials have been isolated from naturally
occurring substances over the years. Our findings indicate
that jujube honey inhibits the initial phase of biofilm formation
and has fungistatic, fungicidal and antibiofilm
potential. This potential is superior to that of most of the
commonly used antifungals. Because biofilms are multifactorial
phenomena, multiple mechanisms that target different
steps in biofilm development are probably involved in the
effects of honey on biofilms. This intriguing observation
may have important clinical implications that could lead
to a new approach for the management of C. albicans
In conclusion, the findings indicate that jujube honey can
inhibit C. albicans biofilms. The significant antifungal activity
of jujube honey suggests that this could serve as a source
of compounds which have a therapeutic potential for the
treatment of Candida-related infections. Further evaluation
in vivo is required to determine whether these findings can
be exploited in treating biofilm-associated candidiasis.