Antifungal effects of ethanolic and aqueous extracts of Vitex agnus-castus against vaginal isolates of Candida albicans

Abstract

Background and Purpose:Vulvovaginal candidiasis is one of the most common infections in female genital organs, which is caused by Candida species. Candida albicans is the causative agent of more than 80% of infections, and the role of non-Candida strains in the disease etiology is less prominent. The expansion of Azoles resistance among C. albicans strains is considered an important medical problem. According to previous studies, Vitex agnus-castus (vitex) has some antimicrobial effects. We aimed to evaluate the anti-fungal effects of aqueous and alcoholic extracts of vitex against clinical vaginal isolates of C. albicans in comparison with fluconazole.Materials and Methods:Gas chromatography-mass spectrometry analysis was performed on vitex to identify its possible bioactive components. Forty C. albicans clinical isolates were identified by using germ tube, chlamydospore production, culture on CHROMagar, and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP). Finally, after the extraction of vitex, drug susceptibility test was carried out according to the clinical laboratory standards institute (CLSI) M27-S4 document guidelines.Results:The major chemical components of vitex leaf as determined by gas chromatography included α-Pinene, isoterpinolene, caryophyllene, and azulene. The minimum inhibitory concentrations (MICs) of aqueous and alcoholic extracts of vitex, as well as fluconazole were within the ranges of 15.62–62.5, 7.81–15.62, and 0.25–8 μg/mL, respectively.Conclusion:Our findings showed that the alcoholic and aqueous extracts of vitex had antifungal activity against clinical isolates of C. albicans. Moreover, the alcoholic extract of vitex and fluconazole were more effective against clinical vaginal isolates of C. albicans compared to the aqueous extract of vitex.

Supplemental Information 1: Anticandidal activity of VAC leaves extract at concentration of 20% by using well diffusion assay.

“…Several of these secondary metabolites belong to important chemical groups such as polyphenols, fatty acids, terpenes, terpenoid, steroids, aldehydes, alcohol, and esters. These results are in agreement with a previous study of Keikha et al (2018), which stated that the VA-C extract was rich in chemical compounds, and the alcoholic extract contained 36 chemical compounds that belong to different chemical groups. Our results showed that the majority of compounds were 4,5-Dichloro-1,3-dioxolan-2-one in both ethanol and methanol, 1H-Indene, 2,3-dihydro-1,1,2,3,3pentamethyl in ethanol, Isobutyl 4-hydroxybenzoate in methanol, and Benzoic acid and 4-hydroxy-in water.…”

“…Moreover, we showed that the inhibitory capacity of the solvents varied significantly in descending order of ethanol, then water, then methanol. These results are in line with a recent study conducted by Keikha et al (2018) who found that VA-C ethanol leaf extract has the highest inhibiting effect against C. albicans isolates than water extract. Our results showed a similarity in the inhibitory effect of all extracts with nystatin (10 mcg) as a positive control against C. albicans, where the inhibitory effect for the positive control was higher than all extracts against C. tropicalis.…”

“…For water and methanol the MIC values are specified at 25 µg/ml which represents dilution 3. Our results are similar to those of Keikha et al (2018) who also found that the ethanol extract of VA-C was more effective than the aqueous extract when the MIC values of ethanol against isolates of Candida species were between 0.78 µg/ml and 1.56 µg/ml, which represent dilution 7 and 8, respectively. The values of the aqueous extract were between 6.25 µg/ml and 1.562 µg/ml, which represent dilutions of 5 and 7, respectively.…”

“…Our results showed that the majority of compounds were 4,5-Dichloro-1,3-dioxolan-2-one in both ethanol and methanol, 1H-Indene, 2,3-dihydro-1,1,2,3,3pentamethyl in ethanol, Isobutyl 4-hydroxybenzoate in methanol, and Benzoic acid and 4-hydroxy-in water. Keikha et al (2018) found that the majority of compounds in the VA-C ethanol extract were a-Pinene, isoterpinolene, caryophyllene, and azulene. The difference in the number of phytochemical compounds may be attributed to the variations among the VA-C genotypes (Karaguzel & Girmen, 2009).…”

Vitex agnus-castus L.: Main Features and Nutraceutical Perspectives

“…Vitex agnus-castus L. leaf essential oil eliminated the cariogenic bacteria Streptococcus mutans (MIC = 15.6 µg/mL) [71]. In another study, alcoholic and aqueous extracts had an antifungal effect on C. albicans isolated from clinical vaginal infections [72]. Vitex agnus-castus L. seed essential oil showed antifungal activity against Candida species (IC 50 = 1.072 mg/mL) and also antioxidant potential [73].…”