Antifungal Activity Testing Services

Creative Biolabs provides antifungal activity testing services for various antifungal agents for our customers all over the world.

Background

Fungi are eukaryotic microorganisms closely related to humans. Fungal growth in food and feed and the resulting release of mycotoxins threaten human health and can even lead to death in acute cases. Dermatophytes are a group of pathogenic fungi that can cause superficial infections of hair, skin, and nails. The genera Microsporum, Trichophyton, and Epidermophyton cause significant disease in humans and other animals.

In healthy people, common forms of mycoses are superficial, cutaneous, or subcutaneous, and in some cases systemic, causing diseases ranging from athlete's foot and nail infections to life-threatening severe disseminated disease. On the other hand, immunocompromised individuals are susceptible to a large number of opportunistic fungal pathogens, causing systemic mycoses. Human fungal diseases can be divided into three categories: toxic reactions to toxins present in certain fungi, allergic reactions to fungal proteins, and infections (mycoses). The application of fungal genomics provides a good opportunity for the development of new antifungal drugs.

Antifungal drugs such as azoles and polyenes can effectively control the infection. However, most of these antifungal agents are known to cause side effects. Therefore, there is a need to find new antifungal agents or preparations that are less toxic and more effective. In recent years, more and more clinical applications using natural products such as plant products and beneficial microorganisms such as probiotics have attracted attention. Probiotics, especially lactic acid bacteria, have been screened for antifungal activity and have shown promising anti-tinea effects. Lactic acid bacteria can produce compounds such as organic acids, enzymes, bacteriocins, and other metabolites of vitamins, some of which are low molecular weight biomolecules that have been isolated and have shown potential to inhibit fungal growth.

Antifungal Activity Testing Services

The standardization of in vitro antimicrobial susceptibility testing by the Clinical and Laboratory Standards Institute (CLSI) and the European Committee on Antimicrobial Susceptibility Testing (EUCAST), as well as the provision of reference methods, are important advances in this field and a major advance in the field of medical mycology.

• Broth/agar dilution test
• Broth microdilution method

Gold standard. Microdilution methods have been more readily adopted for mainstream use in clinical laboratories. The methodology for both yeasts and filamentous fungi is standardized and can be used for any newly discovered antifungal agent.

Minimum inhibitory concentration (MIC)

Minimum fungicidal concentration (MFC)

Minimum effective concentration (MEC)

• Disk diffusion assay
  The disk diffusion method is reproducible, inexpensive, and easy to interpret. Importantly, its results are in excellent taxonomic agreement with the MIC results obtained by the broth microdilution method.
• Gradient Diffusion Strips
  Gradient diffusion strips are another alternative, using inhibition bands to read test results to mark the point where the fungal oval growth intersects the strip, indicating the MIC value. Improved discrimination between true resistance and trailing growth phenomenon. Interpretation is somewhat subjective. Not available for antifungals in development.
• Mycelial Growth Rate Method
• Biofilms Prevention and Treatment Assays

• Determination of Lesion of Plasma Membrane
  Scattergram analysis was performed on each sample to assess morphological changes and the percentage of PI-positive cells was determined by flow cytometry.
• Determination of The Mitochondrial Membrane Potential (MMP)
  Membrane potential kit.
  The changes in MMP in fungi after antifungal treatment were analyzed by laser confocal microscopy and flow cytometry with fluorescent dyes.
• Determination of Mitochondrial ATPase Activity
  The mitochondrial ATPase activity in fungal cells is measured using a Micro-ATPase Assay Kit.
• Determination of Endogenous ROS Production
  Endogenous ROS levels in fungal cells are measured by a fluorometric assay using the fluorescent dye.

Antifungal Drugs Mode of Action

The modes of action of antifungal drugs on fungi include inhibition of ergosterol biosynthesis, inhibition of major components of the plasma membrane, non-competitive inhibition of b-(1,3) -D-glucan synthase, inhibition of nucleic acid biosynthesis, and disruption of spindle and cytoplasmic microtubule functions.

Case Studies

The client provided two small molecule samples. We contracted for the dose-response assay to test the inhibitory effects on the Candida albicans biofilm formation. As shown in Fig.1, the molecule could inhibit biofilm formation.

Fig.1 XTT assay.

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Frequently Asked Questions

Creative Biolabs is positioned to be a global leader in preclinical research in the field of live Biotherapeutics or probiotics. Underpinned by our comprehensive technology platform and a multi-background team of experts, will lead to the discovery of new therapeutic, probiotic, and nutritional approaches to maintain health and treat disease. Please feel free to contact us for more details.

For Research Use Only. Not intended for use in food manufacturing or medical procedures (diagnostics or therapeutics). Do Not Use in Humans.