The tests were conducted in triplicate and repeated on three different days to test the repeatability and reproducibility of the experiments.

The results of this study showed that the E. angustifolia mother tincture combination produced a clear growth inhibition effect on the yeast strains. The tinctures showed an inhibitory effect on all the various Candida strains, with the exception of the repeat test on one strain C.

auris that did not show growth inhibition after 24 hours. auris was however inhibited by the ethanol control. Furthermore, the results showed that the growth inhibition effect decreased after 48 hours. pupurea and E. angustifolia homoeopathic mother tincture combination did yield a noteworthy degree of inhibition of fungal growth against a good percentage of the Candida strains utilized in this study.

This study contributes to the body of antimicrobial research involving E. angustifolia homoeopathic mother tinctures. Files and links 1 pdf Ammaarah Ismail WtmS 2. Open Access. Details Title The antifungal properties of a combination of Echinacea purpurea and Echinacea angustifolia mother tinctures on clinically relevant strains of Candida in vitro.

Creators - without role Ammaarah Ismail. Contributors - without role Tobias George Barnard. Awarding Institution University of Johannesburg; Master of Science MS. Theses and Dissertations Master of Science MS , University of Johannesburg. Identifiers Copyright University of Johannesburg.

Academic Unit Department of Complementary Medicine; Faculty of Health Sciences; University of Johannesburg. Language English. Resource Type Thesis. We use strictly necessary cookies which are required to run this site, as a result, users cannot opt-out of strictly necessary cookies. We also use non-essential cookies, which are used to enhance the user experience through analysis of your usage on this site.

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Cookie Preference Center. Your preferences. Of the 23 mutants, only 16 have a reported function. Follow-up analyses revealed an increase in sonication-associated cell death in the yeasts S. cerevisiae and Cryptococcus neoformans after Echinacea extract treatments.

Furthermore, fluorescence microscopy showed that Echinacea-treated S. cerevisiae was significantly more prone to cell wall damage than non-treated cells. This study further demonstrates the potential of gene deletion arrays to understand natural product antifungal mode of action and provides compelling evidence that the fungal cell wall is a target of Echinacea extracts and may thus explain the utility of this phytomedicine in treating mycoses.

Results show that hexane extracts of Echinacea variably inhibit growth of yeast strains of Saccharomyces cerevisiae, Candida shehata, C. kefyr, C. albicans, C. steatulytica and C. Reference method for broth dilution antifungal susceptibility testing of yeasts, Approved Standard, 2nd ed.

CLSI document MA2, Reference method for broth dilution antifungal susceptibility testing of filamentous fungi, Approved Standard, 3rd ed. Methods for Dilution Antimicrobial Susceptibility Tests for Bacteria That Grow Aerobically, Approved Standard, 9th ed.

CLSI document MA9, Antibacterial activity was evaluated against six Gram-negative Bacteria Pseudomonas aeruginosa ATCC, Escherichia coli ATCC , Salmonella enterica subsp. enterica Typhimurium ATCC, Shigella flexnelli ATCC , Acinetobacter baummani ATCC and Klebsiella pneumoniae ATCC and two Gram- positive bacteria Staphylococcus aureus ATCC and Enterococcus faecalis ATCC Antifungal activity was tested against Cryptococcus gattii R, Cryptococcus gattii ATCC , Cryptococcus neoformans H99, Cryptococcus neoformans ATCC , Candida albicans ATCC , Aspergillus fumigatus clinical strain , Trichophyton inderdigitale ATCC , Trichophyton rubrum ATCC and Microsporum gypseum clinical strain.

The inocula of the tested strains were prepared from grown cultures according to the tested microorganism. The suspensions were prepared in phosphate buffered saline PBS to obtain final concentrations of 1.

Then, µL of each solution was transferred to a sterile well microplate with µL of adequate media Müeller- Hinton for bacteria, Sabouraud dextrose for yeasts and potato dextrose for filamentous fungi. An aliquot of µL of each bacterial or fungi suspension was added to the microplate, in duplicate.

Negative and positive controls were used for each tested strain. The plates were incubated at 35 °C for 24 h bacteria or 72 h fungi , and 28 °C for 7 days for Trichophyton spp. Minimum inhibitory concentration MIC was determined by visual inspection as the lower concentration of the extracts that inhibited the microbial growth for each strain.

The increase in reactive oxygen species ROS production, as well as the intracellular proliferation rate of Cryptococcus gatti after phagocytosis by macrophages in the presence of EP dried extracts were also evaluated.

Fresh media were added every 48 h. Bone marrow-derived macrophages BMDM were collected on day 7 and used for subsequent experiments. Sterile circle coverslips mm were introduced in the bottom of each plate well of well plates of phagocytosis.

Then, the cell concentration was adjusted to 2. Viable yeasts of C. gatti were adjusted to 0. After 24 h, coverlips from 24 well plate from phagocytosis assay were carefully removed, washed with sterile PBS, fixed with ice methanol and stained with Panoptic dye.

Macrophages were counted using optical microscopy and phagocytic capacity was expressed by the percentage of macrophages with internalized fungi.

To investigate the capability of macrophages to control fungal proliferation after phagocytosis, at 24 h after infection, each well was washed with PBS and cells were lysed using μL of sterile distilled water.

Each plate was incubated at 37 ° C for 30 min for complete cell lysis. Then, 50 μL of the lysate was collected and plated on Sabouraud dextrose agar to determine the number of colony forming units CFU. The intracellular proliferation rate was calculated by dividing CFU at 24 hours t 24h by the initial CFU at 3 hours t 3h Ribeiro et al.

Atorvastatin as a promising anticryptococcal agent. Int J Antimicrob Agents. For the determination of reactive oxygen species, the same concentrations EP dried extracts concentrations were tested. The response was determined in a fluorimeter, using excitation and emission wavelengths at nm, and expressed as arbitrary units of fluorescence Ribeiro et al.

The content of caffeic acid derivatives in each sample of EP dried extract was determined by UPLC- DAD in triplicate Oliveira et al. A rapid UPLC method for the simultaneous quantitation of caffeic acid derivatives in dried extracts of Echinacea purpurea. J Chromatogr Sci. The analysis was carried out on a Zorbax Eclipse Plus C 18 column 2.

UV detection was performed at and nm. The evaluation of the antimicrobial activity of EP dried extracts and chlorogenic acid reference standard regarding different strains of microorganisms indicated that the extracts could not inhibit the growth of any tested strain of bacteria.

A slight antifungal activity was evidenced, mainly regarding C. neoformans and C. As demonstrated in Table I , samples 1, 2, 4 and 5 did not inhibit fungal growth at the tested concentrations.

Phagocytosis index was calculated as the percentage of macrophages with internalized fungi, regarding total macrophage number. The higher dose tested led to increased phagocytosis, with a more prominent effect for extracts 3 and 4.

The results from the ROS production assay were expressed as fluorescence arbitrary units Figure 1B. Comparing the infected macrophage group with treated groups, it can be observed that EP dried extracts stimulate ROS production in a dose-dependent way.

The intracellular proliferation rate of C. On the other hand, lower fungal growth inside macrophages was evidenced by extract 3 Figure 1C. FIGURE 1 Fungicidal activity from macrophage EP treated against Cryptococcus gattii. Bars represent the mean ±SD.

BMDM Cg. NI: non-infected. The individual and total contents of caffeic acid derivatives in the samples of EP dried extract are presented in Table II.

It was observed a similar amount of caffeic acid derivatives in the extracts, between 0. In addition, sample 3 presented the highest content of caftaric acid 0. According to the quality reports of suppliers, extract 3 was prepared using the roots of Echinacea purpurea.

In contrast, all other extracts were obtained from aerial parts or aerial parts mixed with roots and stems of the plant, and this difference may be related to the variation in caffeic acid derivatives. Thumbnail TABLE II Content of caffeic acid derivatives in EP dried extracts.

Traditionally, Echinacea purpurea is used for the treatment of respiratory and urinary infections. Sharma et al. demonstrated a significant antibacterial activity of an EP ethanolic extract obtained with aerial parts and roots against respiratory bacteria.

The authors attributed the observed activity to a bactericidal effect of the extract and an anti-inflammatory effect, which could reverse the inflammation caused by these bacteria. On the other hand, samples of EP dried extracts evaluated in the present work did not demonstrate activity against bacteria strains and, as previously mentioned, lack of biological activity may be related to variations on the part of the plant, extraction methods and solvents, or even low contents of vegetal markers in assayed extracts.

Many biological activities were previously reported for caffeic acid derivatives, such as chlorogenic acid, including antimicrobial activity Zheng et al. Extrication process of chlorogenic acid in Crofton weed and antibacterial mechanism of chlorogenic acid on Escherichia coli.

J Environ Biol. Activity of caffeic acid derivatives against Candida albicans biofilm. Bioorg Med Chem Lett. Moreover, Kong et al. In vitro activity of chlorogenic acid against Aspergillus fumigatus biofilm and gliotoxin production.

Exp Therap Med. demonstrated that chlorogenic acid decreased biofilm formation by Aspergillus fumigatus. These reports are consistent with the observed antifungal activity of chlorogenic acid, although this compound did not demonstrate antibacterial activity. gattii ATCC The results obtained in our biological tests may indicate that a possible way by which Echinacea purpurea is effective for the control of infections is an indirect mechanism.

The direct antimicrobial effect appears to be less pronounced, since no significant antibacterial activity was observed and only a slight antifungal activity was verified against the strains evaluated under the conditions of the tests.

is primarily attributed to the ketoalkenes and ketoalkynes abundantly present in the roots. Echinacea - Asteraceae - phototoxicity - polyacetylenes - alkylamides - Candida. Arnason Department of Biology University of Ottawa P.

Box Station A Ottawa ON K1N 6N5 Canada Email: jarnason science. ca Phone: Year Archive Subscribe to RSS Please copy the URL and add it into your RSS Feed Reader.