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. 2022 Sep 27;8(10):1014.
doi: 10.3390/jof8101014.

Assessment of Biofilm Formation by Candida albicans Strains Isolated from Hemocultures and Their Role in Pathogenesis in the Zebrafish Model

Sabi Pokhrel  1 , Nawarat Boonmee  2 , Orawan Tulyaprawat  1 , Sujiraphong Pharkjaksu  1 , Iyarit Thaipisutikul  1 , Phoom Chairatana  1 , Popchai Ngamskulrungroj  1 , Chalermchai Mitrpant  2
Affiliations

Assessment of Biofilm Formation by Candida albicans Strains Isolated from Hemocultures and Their Role in Pathogenesis in the Zebrafish Model

Sabi Pokhrel et al. J Fungi (Basel). .

Abstract

Candida albicans, an opportunistic pathogen, has the ability to form biofilms in the host or within medical devices in the body. Biofilms have been associated with disseminated/invasive disease with increased severity of infection by disrupting the host immune response and prolonging antifungal treatment. In this study, the in vivo virulence of three strains with different biofilm formation strengths, that is, non-, weak-, and strong biofilm formers, was evaluated using the zebrafish model. The survival assay and fungal tissue burden were measured. Biofilm-related gene expressions were also investigated. The survival of zebrafish, inoculated with strong biofilms forming C. albicans,, was significantly shorter than strains without biofilms forming C. albicans. However, there were no statistical differences in the burden of viable colonogenic cell number between the groups of the three strains tested. We observed that the stronger the biofilm formation, the higher up-regulation of biofilm-associated genes. The biofilm-forming strain (140 and 57), injected into zebrafish larvae, possessed a higher level of expression of genes associated with adhesion, attachment, filamentation, and cell proliferation, including eap1, als3, hwp1, bcr1, and mkc1 at 8 h. The results suggested that, despite the difference in genetic background, biofilm formation is an important virulence factor for the pathogenesis of C. albicans. However, the association between biofilm formation strength and in vivo virulence is controversial and needs to be further studied.

Keywords: Candida albicans; biofilm; gene; virulence; zebrafish.

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Conflict of interest statement

The authors declare no conflict of interest. The funder had no any role in the design, data collection, analysis, or interpretation regarding the study or in the manuscript preparation.

Figures

Figure 1
Figure 1
Survival of zebrafish larvae injected with either C. albicans strains 104 (non-biofilm former), 57 (weak biofilm former), or 140 (strong biofilm former). The control group was injected with PBS without an organism. Each experiment was performed in duplicate.
Figure 2
Figure 2
Fungal tissue burden in zebrafish larvae that were infected with yeast cells of C. albicans strains 104, 57, or 140. Each experiment was performed in triplicate. CFU, colony forming unit.
Figure 3
Figure 3
The expression levels of eap1, als3, hwp1, bcr1, and mkc1 in C. albicans strains 104, 57, and 140 infected zebrafish larvae, by time point. Each experimental group of 30 zebrafish was injected with 0.7 OD yeast cells of one of the C. albicans strains. The normalized ratios of expression were calculated by comparison with the level of expression of the act-1 gene in each group of planktonic cells. Each experiment was performed in duplicate. h, hour; **, p < 0.01; ***, p < 0.001; and ****, p < 0.0001.
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