Since ancient times, folk medicine and agro-food science have benefitted from the use of plant derivatives, such as essential oils, to combat different diseases, as well as to preserve food. In Nature, essential oils play a fundamental role in protecting the plant from biotic and abiotic attacks to which it may be subjected. Many researchers have analyzed in detail the modes of action of essential oils and most of their components. The purpose of this brief review is to describe the properties of essential oils, principally as antifungal agents, and their role in blocking cell communication mechanisms, fungal biofilm formation, and mycotoxin production.

Essential Oils and Antifungal Activity

A comprehensive review of the antibacterial, antifungal and antiviral potential of essential oils and their chemical constituents against drug-resistant microbial pathogens

The essential oil extracted from the seeds of dill (Anethum graveolens L.) was demonstrated in this study as a potential source of an eco-friendly antifungal agent. To elucidate the mechanism of the antifungal action further, the effect of the essential oil on the plasma membrane and mitochondria of Aspergillus flavus was investigated. The lesion in the plasma membrane was detected through flow cytometry and further verified through the inhibition of ergosterol synthesis. The essential oil caused morphological changes in the cells of A. flavus and a reduction in the ergosterol quantity. Moreover, mitochondrial membrane potential (MMP), acidification of external medium, and mitochondrial ATPase and dehydrogenase activities were detected. The reactive oxygen species (ROS) accumulation was also examined through fluorometric assay. Exposure to dill oil resulted in an elevation of MMP, and in the suppression of the glucose-induced decrease in external pH at 4 µl/ml. Decreased ATPase and dehydrogenase activities in A. flavus cells were also observed in a dose-dependent manner. The above dysfunctions of the mitochondria caused ROS accumulation in A. flavus. A reduction in cell viability was prevented through the addition of L-cysteine, which indicates that ROS is an important mediator of the antifungal action of dill oil. In summary, the antifungal activity of dill oil results from its ability to disrupt the permeability barrier of the plasma membrane and from the mitochondrial dysfunction-induced ROS accumulation in A. flavus.

/pdf/the-mechanism-of-antifungal-action-of-essential-oil-from-25vkdo3aaz.pdf

The mechanism of antifungal action of essential oil from dill (Anethum graveolens L.) on Aspergillus flavus

The unremitting emergence of multidrug-resistant bacterial pathogens highlights a matching need for new therapeutic options. For example, new carbapenemases such as KPC (class A Klebsiella pneumoniae) and NDM-1 (New Delhi metallo-β-lactamase 1) are surfacing, resulting in almost total resistance to β-lactam antibiotics. Furthermore, resistance is quickly disseminated, not only in the healthcare sector, but also within the community at large, because many resistance determinants are carried on mobile genetic elements readily shared among pathogens. The absence of new antibiotics has led to a growing reliance on older, more toxic drugs such as colistin, but resistance to these is already arising. One approach to combat this growing problem is the use of combination drug antibiotic adjuvant therapy, which potentiates the activity of antibiotics. Here, we review the current situation and discuss potential drug combinations that may increase the potency of antibiotics in the future. Adjuvant therapies include antibiotic combinations, synergy between antibiotics and nonantibiotics, inhibition of resistance and molecules that alter the physiology of antibiotic-insensitive cells, such as those in biofilms. We provide a rationale for these multicomponent strategies, highlighting current research and important considerations for their clinical use and pharmacological properties.

Antibiotic adjuvants: multicomponent anti-infective strategies

Silver nanoparticles have been shown to be detrimental to fungal cells although the mechanism(s) of action have not been clearly established. In this study, we used Candida albicans cells to show that silver nanoparticles exert their antifungal effect through apoptosis. Many studies have shown that the accumulation of reactive oxygen species induces and regulates the induction of apoptosis. Furthermore, hydroxyl radicals are considered an important component of cell death. Therefore, we assumed that hydroxyl radicals were related to apoptosis and the effect of thiourea as a hydroxyl radical scavenger was investigated. We measured the production of reactive oxygen species and investigated whether silver nanoparticles induced the accumulation of hydroxyl radicals. A reduction in the mitochondrial membrane potential shown by flow cytometry analysis and the release of cytochrome c from mitochondria were also verified. In addition, the apoptotic effects of silver nanoparticles were detected by fluorescence microscopy using other confirmed diagnostic markers of yeast apoptosis including phosphatidylserine externalization, DNA and nuclear fragmentation, and the activation of metacaspases. Cells exposed to silver nanoparticles showed increased reactive oxygen species and hydroxyl radical production. All other phenomena of mitochondrial dysfunction and apoptotic features also appeared. The results indicate that silver nanoparticles possess antifungal effects with apoptotic features and we suggest that the hydroxyl radicals generated by silver nanoparticles have a significant role in mitochondrial dysfunctional apoptosis.

https://febs.onlinelibrary.wiley.com/doi/pdf/10.1111/j.1742-4658.2012.08527.x

Silver nanoparticles induce apoptotic cell death in Candida albicans through the increase of hydroxyl radicals.

Plagiochin E, an antifungal active macrocyclic bis(bibenzyl), induced apoptosis in Candida albicans through a metacaspase-dependent apoptotic pathway

Plagiochin E, an antifungal bis(bibenzyl), exerts its antifungal activity through mitochondrial dysfunction-induced reactive oxygen species accumulation in Candida albicans.

Effect of plagiochin E, an antifungal macrocyclic bis (bibenzyl), on cell wall chitin synthesis in Candida albicans

/pdf/effect-of-plagiochin-e-an-antifungal-macrocyclic-bis-4ifexsxuve.pdf

Effect of plagiochin E, an antifungal macrocyclic bis(bibenzyl), on cell wall chitin synthesis in Candida albicans.

The vitro antifungal activity of retigeric acid B (RAB), a pentacyclic triterpenoid from the lichen species Lobaria kurokawae, was evaluated alone and in combination with fluconazole, ketoconazole, and itraconazole against Candida albicans using checkerboard microdilution and time-killing tests. The MICs for RAB against 10 different C. albicans isolates ranged from 8 to 16 μg/ml. A synergistic action of RAB and azole was observed in azole-resistant strains, whereas synergistic or indifferent effects were observed in azole-sensitive strains when interpreted by a separate approach of the fractional inhibitory concentration index and ΔE model (the difference between the predicted and measured fungal growth percentages). In time-killing tests, we used both colony counts and a colorimetric assay to evaluate the combinational antifungal effects of RAB and azoles, which further confirmed their synergistic interactions. These findings suggest that the natural product RAB may play a certain role in increasing the susceptibilities of azole-resistant C. albicans strains.

In Vitro Activities of Retigeric Acid B Alone and in Combination with Azole Antifungal Agents against Candida albicans

Biofilm formation plays a key role in the life cycles and subsistence of many microorganisms. The human fungal pathogen Candida albicans has a high propensity to develop biofilms and resulted resistant to traditional antifungal agents. Biofilms are composed of a mixture of cell types, including yeast, pseudohyphal and hyphal cells, and hyphae are a prominent feature of biofilms. Riccardin D is a macrocyclic bisbibenzyl isolated from the liverwort Dumortiera hirsute in our laboratory. In the present investigation, the XTT [2,3-bis(2-methoxy-4-nitro-5-sulfo-phenyl)-2H-tetrazolium-5-carboxanilide] reduction assay and live/dead cell staining were employed for evaluating the effects of riccardin D on C. albicans biofilms. The results demonstrated that riccardin D can interfere with the biofilm formation. To investigate whether this effect was due to the inhibition of hyphae formation, morphological observation and real-time reverse transcriptase polymerase chain reaction (RT-PCR) were employed for evaluating the effects of riccardin D on the hyphae formation and the expression of hyphae specific genes. The results showed that the hyphae formation was strongly inhibited and the mRNA expression levels of hyphae specific genes were downregulated after riccardin D treatment. We concluded that riccardin D interfered with the biofilm formation of C. albicans through downregulating the expression of hyphae specific genes and inhibiting the formation of hyphae.

Xiu-Zhen Wu

Papers

Plagiochin E, an antifungal active macrocyclic bis(bibenzyl), induced apoptosis in Candida albicans through a metacaspase-dependent apoptotic pathway

Plagiochin E, an antifungal bis(bibenzyl), exerts its antifungal activity through mitochondrial dysfunction-induced reactive oxygen species accumulation in Candida albicans.

Effect of plagiochin E, an antifungal macrocyclic bis(bibenzyl), on cell wall chitin synthesis in Candida albicans.

In Vitro Activities of Retigeric Acid B Alone and in Combination with Azole Antifungal Agents against Candida albicans

The Inhibitory Effect of a Macrocyclic Bisbibenzyl Riccardin D on the Biofilms of Candida albicans