## 4 Discussion  
Our study demonstrated that _Leuconstoc citream_ THY-F51, a lactic acid bacterium isolated from kiwi fruit, exhibited significant antifungal activity against _Candida_ species, the major causative agents of VVC. Additionally, it was shown to effectively suppress _Candida_-induced inflammatory cytokine production and inhibit biofilm formation, which is a key factor in _Candida_ pathogenicity. These findings suggest that _L. citream_ THY-F51 may serve as a next-generation, safe, and promising alternative for patients facing antibiotic resistance, highlighting its potential as a preventive and therapeutic agent for managing VVC, a prevalent infection of the female reproductive tract. Noteworthy findings include the antifungal activity of THY-F51's CFS against _C. albicans_, with MIC and MFC values of 1.25 mg/mL (250 \(\mu\)L/mL) and 2.5 mg/mL (500 \(\mu\)L/mL), respectively (Figure 2, Table 1). These values are comparable to those from antifungal studies using a _Lactobacillus plantarum_ CFS, which reported MIC and MFC values of 200 \(\mu\)L/mL [32]. While _L. plantarum_ is a well-established probiotic strain with confirmed antifungal properties, the antifungal activity of THY-F51 has not been previously studied, making these findings a novel contribution to the field. Furthermore, SEM analysis (Figure 3) revealed significant morphological changes in the _C. albicans_ cell wall, such as deformation and shrinkage, suggesting that the antifungal activity of THY-F51 may be due to metabolites like bacteriocins, organic acids, and pH-altering compounds. These metabolites likely disrupted lipid-protein interactions in the fungal membrane, leading to destabilization and deformation [33]. While various antifungal compounds were evaluated, the exact active agents remain unidentified, highlighting the need for further mechanistic studies and genomic profiling to fully characterize their antifungal potential. The current study demonstrated that the CFS of THY-F51 significantly inhibits fungal biofilm formation in _C. albicans_ KCTC 7270, _C. albicans_ ATCC 10231, and _C. tropicalis_ KCTC 17762, with biofilm formation significantly reduced at 1\(\times\) MIC (1.25 mg/mL) and nearly completely abolished at 2\(\times\) MIC (2 mg/mL), while the control group treated with an MRS medium exhibited enhanced biofilm formation at higher concentrations (Figure 4), indicating the potent anti-biofilm activity of _L. citream_ THY-F51 and its potential as a therapeutic candidate for biofilm-associated fungal infections, in line with a previous study by [PERSON] et al. (2016) [34]. Figure 8: (**a**) The inhibitory effect of THY-F51 on the adhesion of _Candida_ to HeLa cells. (**b**) Acid tolerance test: the survival rate of THY-F51 under vaginal pH (4.0) conditions was measured by colony-forming units (CFUs). \({}^{\star}\), \(p<0.01\); \({}^{\star\star}\), \(p<0.001\). VVC is a condition where inhibiting not only the _Candida_ species but also inflammation plays a crucial therapeutic role [35]. _C. albicans_ adheres to the vaginal epithelium, and the hyphal form invades tissues, causing damage to epithelial cells and triggering immune responses [36]. The host immune system recognizes _Candida_ via pattern recognition receptors (PRRs) such as Toll-like receptors (TLRs) and Dectin-1, which are expressed on the surface of immune cells like dendritic cells and macrophages [37]. Upon recognition of Candida, immune cells initiate the production of inflammatory cytokines such as IL-1B, IL-6, IL-8, and TNF-\(\alpha\), as well as chemotactic factors [38]. During biofilm formation, _Candida_ produces a toxin called candidalysin, which disrupts cell membranes and causes direct damage to host tissues [39]. This leads to the release of damage-associated molecular patterns (DAMPs), which further stimulate the immune response and amplify the inflammatory process, contributing to the release of inflammatory mediators, including TNF-\(\alpha\), IL-1B, IL-6, IL-8, prostaglandandins, histamine, chemokines, MMPs, nitric oxide, and leukotrienes [40].