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Methodologies for in vitro and in vivo evaluation of efficacy of antifungal and antibiofilm agents and surface coatings against fungal biofilms

Authors: M. Ghannoum and More

Publication Year: 2018

PMID: 29992128

Superficial fungal infections of the skin and nails are the most common fungal infections in humans and, although rarely invasive, they can be debilitating, persistent and impose substantial treatment costs 1. In contrast, invasive fungal infections are life threatening, with a higher mortality rate per year than that by malaria, breast or prostate cancer 2. More than 90% of all reported fungal-related deaths (about one million people every year) result from species that belong to one of four genera: CryptococcusCandidaAspergillus and Pneumocystis 2,3.

The most important antifungal agents (antimycotics) clinically used for systemic infections can be subdivided into four main classes: azoles, polyenes, echinocandins and pyrimidine analogues (5-fluorocytosine). In addition, allylamines (terbinafine) are frequently used against superficial fungal infections 4. The rise in azole resistance, echinocandin resistance and cross-resistance to at least 2 antifungal classes (multi-drug resistance: MDR) has been a worrisome trend, mainly in large tertiary and oncology centers. Overall, rates of antifungal resistance and MDR in Candida species and particularly in the emerging human pathogen C. glabrata are increasing 5. More concerning, the newly identified Candida species C. auris has drawn considerable attention as this uncommon species is the first globally emerging fungal pathogen exhibiting MDR to the three major classes of antifungals (azoles, echinocandins and amphotericin B and its lipid formulations) and is characterized by a strong potential for nosocomial transmission 6. In addition to Candida, azole resistance in Aspergillus fumigatus has been reported worldwide, and such resistant isolates can cause invasive infections with high mortality rates 7. Alongside the serious issues presented by classical MDR, there is another important, but less appreciated problem with our current approach to antimicrobial therapy in general. Existing antimicrobial treatments are frequently associated with therapeutic failure even against infections caused by susceptible strains due to intrinsic mechanisms that protect the micro-organisms from the antimicrobial agents, such as the formation of drug-tolerant biofilms. Microbial biofilms consist of dense layers of microorganisms surrounded by an extracellular polymer matrix, which provides biofilm-embedded microorganisms with protection against antimicrobial agents. Most bacteria and fungi exist predominantly in such organized communities in nature and, according to a recent public announcement from the US NIH, biofilms are responsible for more than 80% of human soft- and hard-tissue infections 8. Of more significance, microbial biofilms are thought to result in therapeutic failure and occurrence of resistance 9,10,11,12.

Therefore, the development of accurate susceptibility testing methods for detecting or excluding antifungal resistance, as well as discovery of novel antifungal and antibiofilm agents, are key priorities in medical mycology research. The term ‘antibiofilm agents’ relates to compounds that can inhibit biofilm formation and/or eradicate fungal cells in the biofilm.

To direct advancements in this field, we present in this review an overview of methods for use by investigators who aim to examine:

(i) susceptibility (and resistance) of fungal cultures or biofilms against antifungal or antibiofilm compounds and compound combinations;

(ii) in vivo efficacy of antifungal and antibiofilm compounds and compound combinations; and

(iii) in vitro and in vivo performance of anti-infective coatings and materials to prevent fungal biofilm-related infections.

Several of these topics are already covered in recent guideline-style based reviews 13,14,15,16. We refer to these reviews in the relevant sections and summarize their most important recommendations and guidelines.

Read the Full Report Here.