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Candida albicans causes a variety of clinical manifestations through multiple virulence factors that act simultaneously to overcome the immune system and invade the host tissues. Owing to the limited number of antifungal agents available, new candidiasis therapeutic strategies are required. Previous studies have demonstrated that the metabolites produced by Streptococcus mutans lead to a decrease in the number of Candida cells. Here, for the first time, we evaluated whether the C. albicans cells that survived the pretreatment with S. mutans supernatant can modify their virulence factors and their capability to infect Galleria mellonella larvae. Streptococcus mutans supernatant (SM-S) was obtained by filtering the culture supernatant of this bacterium. Then, C. albicans cells were pretreated with SM-S for 24 h, and the surviving cells were evaluated using in vitro and in vivo assays. The C. albicans pretreated with SM-S showed a significant inhibition of hyphal growth, an altered adhesion pattern, and an impaired capability to form biofilms; however, its proteolytic activity was not affected. In the in vivo assays, C. albicans cells previously exposed to SM-S exhibited a reduced ability to infect G. mellonella and a higher amount of circulating hemocytes. Thus, SM-S could inhibit important virulence factors of C. albicans, which may contribute to the development of new candidiasis therapeutic strategies., (© 2023. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

Amphotericin B (AmB) is the gold standard for antifungal drugs. However, AmB systemic administration is restricted because of its side effects. Here, we report AmB loaded in natural rubber latex (NRL), a sustained delivery system with low toxicity, which stimulates angiogenesis, cell adhesion and accelerates wound healing. Physicochemical characterizations showed that AmB did not bind chemically to the polymeric matrix. Electronic and topographical images showed small crystalline aggregates from AmB crystals on the polymer surface. About 56.6% of AmB was released by the NRL in 120 h. However, 33.6% of this antifungal was delivered in the first 24 h due to the presence of AmB on the polymer surface. The biomaterial's excellent hemo- and cytocompatibility with erythrocytes and human dermal fibroblasts (HDF) confirmed its safety for dermal wound application. Antifungal assay against Candida albicans showed that AmB-NRL presented a dose-dependent behavior with an inhibition halo of 30.0 ± 1.0 mm. Galleria mellonella was employed as an in vivo model for C. albicans infection. Survival rates of 60% were observed following the injection of AmB (0.5 mg.mL -1 ) in G. mellonella larvae infected by C. albicans. Likewise, AmB-NRL (0.5 mg.mL -1 ) presented survival rates of 40%, inferring antifungal activity against fungus. Thus, NRL adequately acts as an AmB-sustained release matrix, which is an exciting approach, since this antifungal is toxic at high concentrations. Our findings suggest that AmB-NRL is an efficient, safe, and reasonably priced ($0.15) dressing for the treatment of cutaneous fungal infections., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023. Published by Elsevier B.V.)

Candida albicans is a pathogen equipped with a variety of commensal and virulence traits that help it colonize the microbiota and invade host tissue during infection. In this study, we investigated the potential anticandidal activity of 3-[2-(4-(4-methoxyphenyl)thiazol-2-yl)hydrazino)]butan-1-ol (MT), a thiazolylhydrazone compound synthesized by our group, and identified it as a promising antifungal agent. The activity of MT was evaluated in vitro and in vivo against C. albicans as well as its ability to inhibit virulence factors. For this, the ability of MT to inhibit the adhesion of C. albicans to human buccal epithelial cells and biofilm formation and filamentation was tested. In addition, the potential in vivo activity of MT was evaluated in murine models of oral candidiasis. Our results confirmed the antifungal activity of MT, with a minimal inhibitory concentration range of 0.5-2 µg/mL. Indeed, MT treatment in vitro decreased the expression of C. albicans genes involved in biofilm formation and morphogenesis and encoding hydrolytic enzymes, which was also confirmed through phenotypic observations. In addition, MT promoted a decrease in the colony forming units recovered from the tongues of mice with oral candidiasis. In this work, we present a potent antivirulence compound that shows potential for candidiasis therapy, especially for topical use., Competing Interests: Conflicts of Interest The authors declare no conflict of interest., (Copyright © 2021 Elsevier Masson SAS. All rights reserved.)

Introduction : Candida  spp. are commensal yeasts capable of causing infections such as superficial, oral, vaginal, or systemic infections. Despite medical advances, the antifungal pharmacopeia remains limited and the development of alternative strategies is needed. Areas covered : We discuss available treatments for Candida spp. infections, highlighting advantages and limitations related to pharmacokinetics, cytotoxicity, and antimicrobial resistance. Moreover, we present new perspectives to improve the activity of the available antifungals, discussing their immunomodulatory potential and advances on drug delivery carriers. New therapeutic approaches are presented including recent synthesized antifungal compounds (Enchochleated-Amphotericin B, tetrazoles, rezafungin, enfumafungin, manogepix and arylamidine); drug repurposing using a diversity of antibacterial, antiviral and non-antimicrobial drugs; combination therapies with different compounds or photodynamic therapy; and innovations based on nano-particulate delivery systems. Expert opinion : With the lack of novel drugs, the available assets must be leveraged to their best advantage through modifications that enhance delivery, efficacy, and solubility. However, these efforts are met with continuous challenges presented by microbes in their infinite plight to resist and survive therapeutic drugs. The pharmacotherapeutic options in development need to focus on new antimicrobial targets. The success of each antimicrobial agent brings strategic insights to the next phased approach in treating Candida  spp. infections.

Background: Antimicrobial photodynamic therapy (aPDT) shows antimicrobial activity on yeast of the genus Candida. In aPDT, the depth at which the light penetrates the tissue is extremely important for the elaboration of the treatment. The aim of this study was to evaluate the action of aPDT on experimental candidiasis and the laser impact in the tissue using Galleria mellonella as the infection model., Methods: G. mellonella larvae were infected with different Candida albicans strains. After 30 min, they were treated with methylene blue-mediated aPDT and a low intensity laser (660 nm). The larvae were incubated at 37 °C for seven days and monitored daily to determine the survival curve, using the Log-rank test (Mantel Cox). To evaluate the distribution of the laser as well as its depth of action in the larva body, the Interactive 3D surface PLOT of Image J was used. The effects of aPDT on the immune system were also evaluated by the quantification of hemocytes in the hemolymph of G. mellonella after 6 h of Candida infection (ANOVA and Tukey's test)., Results: In both the ATCC 18,804 strain and the C. albicans clinical strain 17, aPDT prolonged the survival of the infected G. mellonella larvae by a lethal fungal dose. There was a statistically significant difference between the aPDT and the control groups in the ATCC strain (P = 0.0056). The depth of laser action in the insect body without the photosensitizer was 2.5 mm and 2.4 mm from the cuticle of the larva with the photosensitizer. In the larvae, a uniform distribution of light occurred along 32% of the body length for the group without the photosensitizer and in 39.5% for the group with the photosensitizer. In the immunological analysis, the infection by C. albicans ATCC 18,804 in G. mellonella led to a reduction in the number of hemocytes in the hemolymph. The aPDT and laser treatment induced a slight increase in the number of hemocytes., Conclusion: Both aPDT and laser treatment positively influenced the treatment of experimental candidiasis. G. mellonella larvae were a useful model for the study of light tissue penetration in antimicrobial photodynamic therapy., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Investigation into new therapeutic strategies, such as the use of bacterial isolates with probiotic characteristics, has increased in importance due to the high incidence of Candida albicans and non-albicans Candida infections. This study evaluates Lactobacillus paracasei, Lactobacillus fermentum and Lactobacillus rhamnosus strains as prophylactic and therapeutic agents against infection caused by Candida albicans, Candida glabrata, Candida krusei, and Candida tropicalis in a Galleria mellonella model. Prophylactic treatment provided greater benefits during Candida spp. infection, increasing G. mellonella survival, compared to therapeutic treatment. This study demonstrated that the different Lactobacillus species are potent prophylactic agents of Candida species infection., (© The Author(s) 2018. Published by Oxford University Press on behalf of The International Society for Human and Animal Mycology.)

Candida albicans is the most common human fungal pathogen and can grow as yeast or filaments, depending on the environmental conditions. The filamentous form is of particular interest because it can play a direct role in adherence and pathogenicity. Therefore, the purpose of this study was to evaluate the effects of three clinical strains of Lactobacillus on C. albicans filamentation as well as their probiotic potential in pathogen-host interactions via an experimental candidiasis model study in Galleria mellonella. We used the reference strain Candida albicans ATCC 18804 and three clinical strains of Lactobacillus: L. rhamnosus strain 5.2, L. paracasei strain 20.3, and L. fermentum strain 20.4. First, the capacity of C. albicans to form hyphae was tested in vitro through association with the Lactobacillus strains. After that, we verified the ability of these strains to attenuate experimental candidiasis in a Galleria mellonella model through a survival curve assay. Regarding the filamentation assay, a significant reduction in hyphae formation of up to 57% was observed when C. albicans was incubated in the presence of the Lactobacillus strains, compared to a control group composed of only C. albicans. In addition, when the larvae were pretreated with Lactobacillus spp. prior to C. albicans infection, the survival rate of G. mellonela increased in all experimental groups. We concluded that Lactobacillus influences the growth and expression C. albicans virulence factors, which may interfere with the pathogenicity of these microorganisms.

Candida albicans and Candida tropicalis are commensal microorganisms occurring in the oral cavity of approximately 50%-70% of healthy individuals. However, these microbes can become pathogenic through changes in the environment or weakened host immune system. Thus, the aim of this investigation was to evaluate the interaction between species of the genus Candida in the biofilm formation, filamentation, gene expression and virulence in Galleria mellonella. Coincubation of C. albicans with C. tropicalis cells after 48 h resulted in significant reduction of biofilm formation by decreasing viable cell counts, metabolic activity and hyphal growth. The C. albicans genes (BCR1, CPH1, EFG1, UME6, HWP1, ALS3, SAP5 and PLB2) were quantified by quantitative real-time polymerase chain reaction and most of genes were downregulated. Regarding in vivo assay, the groups that the larvae received C. albicans and C. tropicalis had a significant survival increase compared to the control group of C. albicans (P = 0.0001) in agreement with the in vitro results. In conclusion, C. tropicalis colonization was associated with a decrease in the growth of C. albicans, suggesting an antagonistic relation between these two species. Therefore, C. tropicalis by reducing C. albicans virulence profile may limit the ability of this pathogenic fungus to cause infection.

Due to the growing number of multi-resistant Candida spp., adjuvant treatments that may help combat these fungal pathogens are relevant and useful. This study evaluated the immunomodulation and anti-Candida activity of Lactobacillus rhamnosus (LR), Lactobacillus acidophilus and Lactobacillus paracasei suspensions, either single- or multiple-strain, in mouse macrophages (RAW 264.7) and Galleria mellonella (GM). Mouse macrophages were activated by different lactobacilli suspensions and challenged with C. albicans (CA). Tumor necrosis factor (TNF)-α, interleukin IL-1β, IL-6 and IL-17 production and cell viability were investigated. LR was the best suspension for stimulating all evaluated cytokines and thus was used in subsequent in vivo assays. Two C. albicans clinical strains, CA21 and CA60, were then added to the GM assays to further confirm the results. LR suspension was injected into the larvae 24 h before challenging with CA. Survival curve, CFU per larva and hemocytes were counted. In the GM, the LR suspension increased the survival rate and hemocyte counts and decreased the CFU per larva counts for all groups. Lactobacilli suspensions presented strain-dependent immunomodulation; however, single suspensions showed better results. Anti-Candida activity was demonstrated by decreased Candida counts in the GM with the use of LR., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

In this study, we evaluated the interactions between Candida albicans, Candida krusei and Candida glabrata in mixed infections. Initially, these interactions were studied in biofilms formed in vitro. CFU/mL values of C. albicans were lower in mixed biofilms when compared to the single biofilms, verifying 77% and 89% of C. albicans reduction when this species was associated with C. glabrata and C. krusei, respectively. After that, we expanded this study for in vivo host models of experimental candidiasis. G. mellonella larvae were inoculated with monotypic and heterotypic Candida suspensions for analysis of survival rate and quantification of fungal cells in the haemolymph. In the groups with single infections, 100% of the larvae died within 18 h after infection with C. albicans. However, interaction groups achieved 100% mortality after 72 h of infection by C. albicans-C. glabrata and 96 h of infection by C. albicans-C. krusei. C. albicans CFU/mL values from larvae hemolymph were lower in the interacting groups compared with the monoespecies group after 12 h of infection. In addition, immunosuppressed mice were also inoculated with monotypic and heterotypic microbial suspensions to induce oral candidiasis. C. albicans CFU/mL values recovered from oral cavity of mice were higher in the group with single infection by C. albicans than the groups with mixed infections by C. albicans-C. glabrata and C. albicans-C. krusei. Moreover, the group with single infection by C. albicans had a higher degree of hyphae and epithelial changes in the tongue dorsum than the groups with mixed infections. We concluded that single infections by C. albicans were more harmful for animal models than mixed infections with non-albicans species, suggesting that C. albicans establish competitive interactions with C. krusei and C. glabrata during biofilm formation and development of experimental candidiasis.

The number of studies using G. mellonella as a model host for human pathogens has increased significantly in the last few years. Important studies were published from different countries for evaluating the pathogenesis of bacterial and fungal infections and for exploring the host defenses against pathogens. Therefore, standardized conditions for the use of G. melonella larvae need to be established. Recent research showed that the deprivation of G. mellonella larvae of food during the experiment caused a reduction in immune responses and an increased susceptibility to infection, suggesting that incubating of larvae in the presence or absence of nutrition may affect the results and comparisons among different laboratories.