Virgin Coconut Oil Inhibits Candida Albicans Growth In-vitro

Umi Fitriyani

, Meizly Andina

Faculty of Medicine, University of Muhammadiyah Sumatera Utara, Medan, Indonesia

Keywords: Virgin coconut oil, Candida albicans.

Abstract: Candida albicans is the commensal organism act as a normal flora of the human body that can turn into

pathogens. Virgin Coconut Oil (VCO) is an oil obtained from coconut meat (Cocos nucifera l.). The

purpose of this research is to obtain the inhibitory power of pure coconut oil (virgin coconut oil) to Candida

albicans growth in-vitro. Preparation of VCO was set up in four concentrations, i.e. VCO 100%, 50%, 25%,

12.5%, fluconazole and distilled water as the control. The VCO antifungal test is performed through

diffusion using the disc diffusion method (Kirby and Bauer test). VCO 50% has an average resistor of 5 cm

larger than the average inhibition power of VCO 100% that is 4.5 cm, 1 cm on VCO 25% and 0.7 cm on

VCO 12.5%. The inhibitory power of the fluconazole drug discs showed significantly different inhibitory

power results (P-value <0.05) compared with VCO with 100%, 50%, 25%, 12.5% concentration, and

distilled water. While the results of Mann Whitney-Test all VCO concentrations did not show differences in

inhibitory results on the growth of C. albicans (P> 0.05), except in VCO concentrations 50% compared with

12.5% and distilled water (p <0.05). All VCO concentrations can inhibit the growth of Candida albicans in-

vitro starting from concentration but the inhibitory power of the VCO test group is not as effective as the

inhibitory power of the fluconazole drug disc.

1 INTRODUCTION

Candidiasis is a fungal disease that is caused by

Candida sp. The most common cause is Candida

albicans species. Candida albicans infection may

affect the mouth, vagina, skin, nails, respiratory

tract, and at some point can also cause septicemia,

endocarditis, or meningitis if not adequately treated

(Kuswadji, 2007).

Candida albicans is one of the commensal

organisms that act as a normal flora of the human

body especially in the gastrointestinal mucous

membranes (24%) and the vaginal mucosa (5-11%)

and this species is in normal state, therefore it is

harmless to our body (Kayser, 2010). However, in

the event of a disruption such as a weak immune

system (in the case of HIV-AIDS), Candida

albicans, originally a normal flora in the body, may

become pathogenic, causing various diseases, such as

vulvovaginal candidiasis. Based on the results of the

study, three out of four women had at least once

experienced vulvovaginal candidiasis in their lives

(Hidalgo, 2012).

Pure coconut oil or Virgin Coconut Oil (VCO)

is an oil obtained from old and fresh coconut meat

(Cocos nucifera l.) processed by squeezing with or

without water addition, with or without heating that

is not more than 60oC and safe for human

consumption (SNI, 2008). Based on the research,

the content of lauric acid and capric acid in VCO

can kill C. albicans by destroying the C. albicans

plasma membrane and making its cytoplasm

shattered and shrunk. Therefore, the VCO is

possible when used as an infectious treatment

caused by Candida albicans that infects the skin

and mucosa, and is also possible to be used as an

antibiotic therapy for long periods of time

(Bergsson, 2001). Another research results on the

antimicrobial power of VCO indicate that VCO has

minimum inhibitory level and minimum killing rate

of 25% by dilution and diffusion method

(Nurjannah, 2012).

Based on the results of research conducted by

Kabara (2005) showed that lauric acid contained in

VCO has a bacteriostatic effect on gram-positive

bacteria and also Candida albicans fungi. Where

the researcher used fatty acid and its derivatives as

antimicrobial substances tested on 9 gram-positive

bacteria, i.e. S. aureus, S. epidermidis,

Streptococcus beta-hemolytic group A,

Fitriyani, U. and Andina, M.

Virgin Coconut Oil Inhibits Candida Albicans Growth In-vitro.

DOI: 10.5220/0008790100230026

In Proceedings of the 2nd Syiah Kuala International Conference on Medicine and Health Sciences (SKIC-MHS 2018), pages 23-26

ISBN: 978-989-758-438-1

Streptococcus beta-hemolytic group non-A,

Streptococci group D, Corynebacteria,

Micrococcus sp., Nocardia asteroides,

Pneumococci, and also Candida albicans. In the

treatment of Candida albicans, the result obtained

was the lauric acid having a minimum inhibitory

level of 2.49 micromol/ml, the capric acid had a

minimum inhibitory level of 2.9 micromol/ml,

while the caprylic acid had no minimum inhibitory

content.

According to Bergsson (2001), the sensitivity

of Candida albicans to fatty acids and some of its

monoglycerides tested by short-acting inactivation,

and ultrathin were studied using an electron

transmission microscope (TEM) after being given a

capric acid. The results showed that the capric acid

(C-10) caused the quickest and most effective

killings of all three C. albicans strains tested,

leaving the cytoplasm in an irregular and shrunken

state due to the disruption or destruction of the

Candida albicans plasma membrane. Lauric acid

(C-12), is the most active at lower concentrations

and after incubation time is longer. Here's a picture

of electron microscope Candida albicans

morphology after being given capric acid.

The objective of this research is to see the

inhibitory power of virgin coconut oil to the growth

of Candida albicans in-vitro.

2 METHOD

The type of this research is laboratory

experiments. This study examined the minimum

inhibitory levels of virgin coconut oil (Virgin

Coconut Oil) on the growth of Candida albicans in

vitro. The research was conducted at the

Microbiology Laboratory, Faculty of Medicine,

University of North Sumatera. The sample of this

study is the culture of Candida albicans fungus

taken from the Microbiology Laboratory, University

of North Sumatera.

The culture of Candida albicans is done by

making the germ suspension made by taking the

culture result (+) with osse, then diluted with 0.9%

NaCl sterile and adjusted to 0.5 McFarland solution.

The germ solution is taken with a sterile cotton

swab, emphasized on the edge of the tube until it

does not drip when removed. Then the cotton swab

was evenly applied on the surface of the Saboraud

Dextrose Agar medium and waited until it dried.

After drying, the disc to be tested is taken with

sterile tweezers and placed on Saboraud Dextrose

Agar media for 24 media. Then incubated for 24

hours at 37

VCO preparations were made into four

concentrations. A total of 12 sterile 3 ml volume

reaction vessels were provided for 3 series VCO

dilutions. For each series of dilutions prepared 4

sterile reaction tubes, then numbered from 1 to 4. As

much as 2 ml VCO is inserted in tube 1 (for 100%

VCO concentration). Then as much as 1 ml VCO is

inserted in tube no.2 and mixed with tween 80 as

much as 1 ml (total volume 2 ml) then stirred until

homogeneous (for making VCO concentration

50%). The VCO has then added as much as 0.5 ml

on tube number 3, then mixed with tween 80 as

much as 1.5 ml, then mixed until homogeneous (to

make VCO concentration 25%). Then as much as

VCO 0.25 is inserted into tube number 4, then mixed

with tween 80 as much as 1.75 ml (for making VCO

concentration 12.5%). Furthermore, blank disc paper

is inserted into each test material cylinder (100%

VCO, 50%, 25%, 12.5%, distilled water) for 10

minutes for the solution to be absorbed into the disc

paper properly.

3 RESULT

The VCO antifungal test is performed by

diffusion using the disc diffusion method (Kirby and

Bauer test). Each sabouraud dextrose agar medium

that has been planted with Candida albicans affixed

to each paper disc containing 100%, 50%, 25%,

12.5% VCO, distilled water, and fluconazole drug

solutions. The preparation is made up of 4 media

and each medium is repeated 4 times.

Table 1: The inhibitory power of all test groups on the

growth of Candida albicans in-vitro

Group

Mean inhibitory

zone diameter

(

)

Standard

deviation

(

)

Fluconazole 17.7 1.78

VCO 100% 4.5 6.22

VCO 50% 5 1.76

VCO 25% 1 2.23

VCO 12.5% 0.7 1.56

Distilled water 0 0

Then all treated media were incubated for 24

hours at 37 ° C in the incubator. Then perform the

drag zone measurements by using the sliding term

SKIC-MHS 2018 - The 2nd Syiah Kuala International Conference on Medicine and Health Sciences

on the clear area that occurs around the hole and

measured with the measuring paper. The clear area

is the diameter of the growing resistance of the

fungus being tested. Then interpret the results of the

measurement of the clear zone.

The result of the inhibitory zone diameter of all

VCO concentrations above is then analyzed by using

SPSS with the initial stage determining the

normality and homogeneity of the data. From the

normality and homogeneity test, the result is that the

data distribution is not normal (p> 0.05).

Furthermore, non-parametric analysis of the Kruskal

Wallis method was performed. The results obtained

are p <0.05 (ie 0.006), this indicates that all the test

groups there is no significant difference.

Mann Whitney-Test was then performed by

comparing each test group. Then the results obtained

are Fluconazole drug disc's inhibitory effect showed

significantly different inhibitory power results (P

<0.05) compared with VCO 100%, 50%, 25%,

12.5%, and distilled water. While the results of

Mann Whitney-Test all VCO concentrations did not

show differences in inhibitory results on the growth

of C. albicans (P> 0.05), except in VCO

concentrations 50% compared with VCO 12.5% and

distilled water (p <0.05) (Table 2).

Table 2: Differences of inhibitory zone results on the

growth of C. albicans.

Group Comparison P-value

Flukonazol

VCO 100% 0.020

<0.05

)

VCO 50% 0.020

<0.05

)

VCO 25% 0.018

<0.05

)

VCO 12,5 % 0.018

<0.05

)

Distilled wate

0.014 (p<0.05)

VCO 100%

VCO 50% 1.000 (p>0.05)

VCO 25% 0.321 (p>0.05)

VCO 12,5 % 0.321

>0.05

)

Distilled wate

0.131

>0.05

)

VCO 50 %

VCO 25% 0.069

>0.05

)

VCO 12,5 % 0.037 (p<0.05)

Distilled wate

0.013 (p<0.05)

VCO 25%

VCO 12,5 % 0.850 (p>0.05)

Distilled wate

0.317

>0.05

)

VCO 12,5 % Distilled wate

0.317

>0.05

)

4 DISCUSSION

VCO concentrations of 100%, 50%, 25%, and

12.5% on average can inhibit the growth of Candida

albicans fungi. The results obtained are in

accordance with the results of research conducted by

Diana,(2009) who also conducted research on the

antifungal effects of VCO against C. albicans. Based

on the results he obtained, the Minimum Depression

Level (KHM) VCO 100% was 14 mm, VCO 50%:

11 mm, VCO 25%: 10 mm and VCO 12.5%: 8mm.

Other studies have also been conducted by

Nurjannah (2012) about the antimicrobial power of

VCO against C. albicans with the tube dilution

method. The results show that VCO has anti mycotic

power against C. albicans with Minimum Fungicidal

Concentrations (MFCs) and Minimum Inhibitory

Concentrations (MICs) are 25%.

However, other studies show that VCO cannot

inhibit the growth of C. albicans in-vitro (Yulian,

2007) (S.S Dewi, T Aryadi, 2010). According to

research the factors that influence is due to the poor

solubility of VCO in the air (Yulian, 2007). Because

in making the concentration of VCO that is by

mixing VCO with tween 80 and distilled water with

ratio 1:1:2.

The use of tween 80 in this study was as a

solvent VCO in the manufacture of concentrations.

Tween 80 is a nonionic surfactant and emulsifier

derived from polyethoxylated sorbitan and oleic

acid. Surfactants are molecules that have hydrophilic

groups and lipophilic groups. Can dissolve in the

form of air or oil. Tween 80 can lower the voltage

between drugs and medium and various micelles.

This building will be carried away by micelles

dissolved in the medium (Zulkarnain, 2008)

In this study, four repetitions were conducted, it

aims to estimate the variations of the experimental

error, to estimate the standard error of the treatment

rate, to improve the test provision, to extend the

precision of the experimental conclusions through

the selection and use of experimental units which is

more varied. If the number of replications increases,

then the alleged population mean value through the

midpoint of the observed treatment becomes more

thorough. The research error is a measure of

diversity among all observations derived from the

experimental units that received the same treatment

(Raupong, 2011).

5 CONCLUSIONS

All VCO concentrations can inhibit the growth of

Candida albicans in-vitro from VCO concentrations

of 12.5% to 100% VCO. However, the inhibitory

power of the VCO test group is not as effective as

the inhibitory power of the fluconazole drug disc.

Virgin Coconut Oil Inhibits Candida Albicans Growth In-vitro

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SKIC-MHS 2018 - The 2nd Syiah Kuala International Conference on Medicine and Health Sciences