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Fig 1.

Point inoculation of C. albicans isolates.

(a) MTCC-3017, (b) C. albicans M-207, (c) C. albicans S-470 on TSA medium at 16 h.

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Fig 1 Expand

Fig 2.

Growth of C. albicans clinical isolates on FBS and non-FBS coated 96 well microtiter plate in TSB medium.

All values are expressed as mean and standard deviation. The experiment was performed in triplicate.

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Fig 3.

Growth profile of C. albicans isolates in TSB medium for 0–72 h.

All values are expressed as mean and standard deviation. The experiment was performed in triplicate.

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Fig 3 Expand

Fig 4.

Antimicrobial activity of aqueous extracts of garlic, clove, and Indian gooseberry with conventional antimycotics.

(1) Clove (43mg), (2) Garlic (200mg), (3) Indian Gooseberry (86mg) with (a) (4) Fluconazole (25mcg) (control) and (b) (4) Caspofungin (8mcg) (control) for (A) C. albicans M-207, (B) C. albicans S-470. Concentrations are expressed as dry weight measurements.

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Table 1.

Zone of inhibition for aqueous extracts of garlic, clove, Indian gooseberry, and standard antimycotic discs of fluconazole and caspofungin by disc diffusion method.

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Fig 5.

Antimicrobial activity of aqueous garlic extract.

(a) garlic extract at 0.25mg—1.25mg (b) garlic extract at 2.5 mg– 200 mg (Neat) on MHA for (A) C. albicans M-207 & (B) C. albicans S-470. Concentrations expressed as dry weight measurements.

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Fig 6.

Antimicrobial activity of aqueous extracts of garlic, clove, and Indian gooseberry on 96 well microtiter plate.

(A) C. albicans M-207, (B) C. albicans S-470 at various concentrations and different time intervals by growth absorbance. All values are expressed as mean and standard deviation. The experiment was performed in triplicate. Concentrations are expressed as dry weight measurements.

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Fig 6 Expand

Fig 7.

Panel A: Antimicrobial activity of aqueous extracts of garlic, clove, and Indian gooseberry by MTT Assay. (A) C. albicans M-207, (B) C. albicans S-470 at different concentrations and at different time intervals. The values are expressed as mean and standard deviation. The assay was performed in triplicate. Concentrations are expressed as dry weight measurements. Panel B: Antimicrobial activity of aqueous extracts of garlic against preformed C. albicans biofilm by MTT Assay. (A) 24 h preformed biofilm of C. albicans M-207, (B) 24 h preformed biofilm of C. albicans S-470, (C) 48 h preformed biofilm of C. albicans M-207, (D) 48 h preformed biofilm of C. albicans S-470.

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Fig 8.

Glass slides immersed in glass Petri plate containing C. albicans treated with garlic, clove, and Indian gooseberry.

(a) C. albicans Control, (b) Aqueous garlic treated, (c) Aqueous clove treated, (d) Aqueous Indian gooseberry treated (A) C. albicans M-207 and (B) C. albicans S-470.

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Fig 9.

HPTLC results of garlic, clove, and Indian gooseberry.

(A) Garlic extract derivatized with ninhydrin—Track 1: Garlic (Aq. Extract); Track 2: Alliin standard (Rf: 0.38), (B) Clove extract derivatized with ferric chloride—Track 1: Clove (Aq. Extract); Track 2: Ellagic acid standard (Rf: 0.3), (C) Indian Gooseberry extract derivatized with ferric chloride—Track 1: Indian Gooseberry (Aq. Extract); Track 2: Gallic acid standard (Rf: 0.82).

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Fig 10.

LC-MS spectra for crude fresh aqueous extract of garlic, clove, and Indian gooseberry.

LC-MS measured at 210 nm, 275 nm, and 254 nm respectively.

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Fig 11.

Microscopic images of crystal violet stained Candida biofilm on a glass slide.

Images captured at 100X magnification at different time intervals (a) 1 h, (b) 3 h, (c) 12 h, (d) 24 h, (e) 48 h, (f) 72 h, (g) 96 h (h) 120 h for (A) C. albicans M-207 control, (B) Aqueous garlic treated C. albicans M-207, (C) C. albicans S-470 control, (D) Aqueous garlic treated C. albicans S-470, (E) Aqueous clove treated C. albicans S-470, (F) Aqueous Indian gooseberry treated C. albicans S-470.

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Fig 12.

Phase-contrast microscopic images in 96 well microtiter plate.

Images captured at 20x magnification at different time intervals (a) 1 h, (b) 3 h, (c) 6 h, (d) 12 h, (e) 24 h, (f) 48 h, (g) 72 h (h) 96 h (i) 120 h for (A) C. albicans M-207 control, (B) Aqueous garlic treated C. albicans M-207 (1mg), (C) C. albicans S-470 control, (D) Aqueous garlic treated C. albicans S-470 (1.25mg), (E) Aqueous clove treated C. albicans S-470 (0.215mg), (F) Aqueous Indian gooseberry treated C. albicans S-470 (0.537mg). Concentrations are expressed as dry weight measurements.

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Fig 13.

Fluorescence images of C. albicans stained with calcofluor white.

Panel A: Images captured at different time intervals (a) 3 h, (b) 6 h, (c) 12 h, (d) 24 h for (A) C. albicans M-207 control, (B) Aqueous garlic treated C. albicans M-207 (1mg), (C) C. albicans S-470 control, (D) Aqueous garlic treated C. albicans S-470 (1mg), (E) Aqueous clove treated C. albicans S-470 (0.215mg), (F) Aqueous Indian gooseberry treated C. albicans S-470 (0.537mg). All images were captured at 40x magnification. Concentrations are expressed as dry weight measurements. Panel B: Quantification of fluorescence intensity of C. albicans M-207 and S-470 treated with garlic, clove, and gooseberry at different time intervals analyzed by the fluorescence of the entire image using ImageJ.

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