JOURNAL OF MEDICINAL FOOD

J Med Food 16 (6) 2013, 499–503

# Mary Ann Liebert, Inc., and Korean Society of Food Science and Nutrition
DOI: 10.1089/jmf.2012.0066

In Vitro and In Vivo Effects of Two Coconut Oils in Comparison

to Monolaurin on Staphylococcus aureus: Rodent Studies
Vijaya Manohar,1 Bobby Echard,1 Nicholas Perricone,2 Cass Ingram,3 Mary Enig,4
Debasis Bagchi,5 and Harry G. Preuss1
1
Department of Biochemistry, Georgetown University Medical Center, Washington, District of Columbia, USA.
2
Michigan State University College of Human Medicine, East Lansing, Michigan, USA.
3
North American Herb And Spice, Buffalo Grove, Illinois, USA.
4
Enig Associates, Inc., Bethesda, Maryland, USA.
5
Department of Pharmacological & Pharmaceutical Sciences, University of Houston, Houston, Texas, USA.

ABSTRACT Since monolaurin, a monoglyceride formed in the human body in small quantities, has proven effective both
in vitro and in vivo against certain strains of Staphylococcus aureus, an important question arises whether consuming a
substance high in lauric acid content, such as coconut oil could increase intrinsic monolaurin production to levels that would
be successful in overcoming staphylococcal and other microbial invaders. Both a cup plate method and a microdilution broth
culture system were employed to test bacteriostatic and bactericidal effects of the test agents in vitro. To test effectiveness
in vivo, female C3H/he mice (10–12 per group) were orally administered sterile saline (regular control), vancomycin (positive
control), aqueous monolaurin, or two varieties of coconut oil (refined, bleached, deodorized coconut oil and virgin coconut oil)
for 1 week before bacterial challenge and 30 days after. A final group received both monolaurin and vancomycin. In contrast
to monolaurin, the coconut oils did not show bactericidal activity in vitro. In vivo, the groups receiving vancomycin,
monolaurin, or the combination showed some protection—50–70% survival, whereas the protection from the coconut oils
were virtually the same as control—0–16% survival. Although we did not find that the two coconut oils are helpful to
overcome S. aureus infections, we corroborated earlier studies showing the ability of monolaurin to do such.

KEY WORDS:  antibacterial  coconut oil  monolaurin

INTRODUCTION refined, bleached, deodorized (RBD) coconut oil (contain-

ing lauric acid approximating 50%), and a virgin coconut oil
M onolaurin, a monoglyceride formed in the human
body in small quantities, has proven effective both in
vitro and in vivo against certain strains of Staphylococcus
containing lauric acid at roughly the same level. As a first
approximation, we also examined, the influences of the
two coconut oil preparations in vitro on two strains of
aureus.1–6 Studies performed in vivo have suggested that
S. aureus, one claimed to be multidrug resistant, before
exogenous monolaurin could essentially be as effective as
launching in vivo experiments. We found that in contrast to
vancomycin.5 The latter scenario leads to an important
pure monolaurin the two brands of coconut oil by in vitro
question: would consuming a substance high in lauric acid
means and given orally to living mice were essentially
content, such as coconut oil increase intrinsic monolaurin
ineffectual against the Staphylococcal organism and the
production to levels that would be successful in overcoming
infection caused by it.
staphylococcal and other microbial invaders4? If so, foods
containing coconut oils in sufficient quantities might become MATERIALS AND METHODS
effective antimicrobial protective agents.
We designed the current study to examine the ability of In vitro studies
two different coconut oils given orally to protect mice Initially a cup plate method was employed to screen all
against a fatal form of S. aureus. The two forms were the test agents, including the antibiotics.7 Molten nutrient
agar cooled to 45C was seeded with exponentially growing
S. aureus cultures and poured into Petri dishes. After the
Manuscript received 27 February 2012. Revision accepted 30 September 2012
media was hardened, cups were made into the agar using a
Address correspondence to: Harry G. Preuss, MD, MACN, CNS, Department of Bio- sterile borer. The wells were filled with the test reagents—
chemistry, Georgetown University Medical Center, Basic Science Building, Rm. 231 B,
3900 Reservoir Rd. NW, Washington, DC 20057, USA, E-mail: preusshg@
0.l mL containing various concentrations of material under
georgetown.edu examination or the solvent control. The plates were

499
500 MANOHAR ET AL.

incubated at 37C for 24 to 48 + h. Antibiotics examined Table 1. Screening of Antibiotics

included; amoxicillin, penicillin, streptomycin, methicillin, Against Staphylococcus aureus Strains
vancomycin, as well as test agents. RBD and virgin coconut
Agents tested, S. aureus BAA 42 S. aureus BAA 14154
oil and monolaurin (glycerol ester of lauric acid) were the lg/mL MIC MBC MIC MBC
test agents. Absolute ethanol was the solvent control. Test
organisms were; S. aureus ATCC strain #BAA42 and Vancomycin 2.5 5.0 25.00 30.00
ATCC strain #14154. Amoxycillin 5.0 5.0 25.00 25.00
Methicillin 2.5 2.5 20.00 25.00
The zone of inhibition was measured to assess the mini- Penicillin 5.0 5.0 30.00 35.00
mum inhibitory concentration (bacteriostatic) at 24 h and Streptomycin 5.0 5.0 30.00 40.00
minimum bactericidal concentrations required at the end of Monolaurin 1.25 2.5 10.00 15.00
48 + h of incubation. Virgin coconut oil was obtained from Coconut oil No apparent killing
North American Herb and Spice (NAHS), Waukeegan, IL, (RBD)
and RBD from San Pablo Manufacturing Corporation, San Ethanol No apparent killing
Pablo City, Philippines. MIC, minimum inhibitory concentration; MBC, minimum bactericidal
A microdilution broth culture system (1 mL) was also concentration; RBD, refined, bleached, deodorized.
employed to test the bacteriastatic (24 h) and bactericidal
(48 + h) effects of the test agents.6,8 Serial dilutions of an-
tibiotics, the two oils under investigation and monolaurin kidney homogenates on SG agar plates further tested the
were made in nutrient broth and inoculated with exponen- renal burden of S. aureus.
tially growing (18 h old) bacteria (6 · 106 cfu). Appropriate
solvent controls were also included. Cultures were incu- RESULTS
bated at 37C on a rotary shaker for 5 days. Aliquots from
the culture were drawn after 24, 48, and 72 h, and plated on In vitro
nutrient agar plates. Plates were incubated in a 37C incu- Using the cup plate method,3 monolaurin produced a clear
bator for the appearance of colonies. area in culture in a dose-concentrated manner (data not shown).
The RBD showed virtually no area of clearing, while the
In vivo studies NAHS coconut oil showed some clearing (data not shown).
Using the microdilution method, initial screening of the
Female C3H/hej mice (15–18 g) were obtained from Ta- panel of antibiotics against the two strains of S. aureus re-
conic Farms (Germantown, NY, USA). The animals, main- vealed that the strain 14154 is in general more resistant and
tained in a controlled environment at 25C with a 12 h light required higher concentrations of antibiotics and mono-
and 12 h dark cycle, were acclimatized for 3–5 days before laurin to be effective (Table 1).
use. The mice were housed in groups of five, fed commercial Figure 1 depicts the ability of monolaurin at different
rodent pellets, and given water ad libitum throughout the concentrations (starting at 2.5 lg/mL) to destroy staphylo-
experiments. The protocol for the in vivo investigation was coccus in vitro, whereas no similar effects were seen with
approved by the Animal Welfare Board at Georgetown RBD coconut oil even at higher concentrations (Fig. 2).
University Medical Center (Washington, DC, USA).
The optimal conditions for murine bacteremia model and In vivo
its treatment by vancomycin were established previously.8
Six groups of 12 mice were injected with 0.75 mL sterile
Twelve animals in each group (single exception was group
saline containing the BAA 14154 strain. The only exception
receiving both monolaurin/vancomycin containing 10 mice)
in numbers was the group receiving the combination of
were orally administered with doses of aqueous monolaurin
monolaurin and vancomycin that contained 10 mice. In the
(5.00 mg in 0.75 mL of sterile saline) and coconut oil
12 receiving olive oil as baseline, the mice died within 8
(0.75 mL) for a week before infection, and this regimen was
days with the exception of one mouse (Fig. 3). Seven of
to continue until the termination of the experiments (30 days
those receiving vancomycin survived 30 days and upon
later). The control groups were gavaged with either 0.75 mL
gross inspection and culture of the kidneys were found free
sterile saline (Regular Control) or 400 lg of vancomycin in
of infection. Examining the results of the mice receiving the
0.75 mL sterile saline (Positive Control). All groups were
natural products, six receiving monolaurin survived, as did
eventually injected intraperitoneally with 25 · 106 bacterial
seven ingesting the combination of monolaurin and vanco-
particles in 0.1 mL of sterile PBS containing 5% mucin. A
mycin. In contrast, none survived that received RBD coco-
new needle and syringe were used for each injection.
nut oil and two survived that got concentrated virgin
The mice were monitored twice daily postinjection with
coconut oil. Again, all survivors were free of staphylococcus
the bacteria until the completion of the experiment. The
in their kidneys after 30 days.
body weight of each mouse was recorded daily. Animals
exhibiting extreme distress, such as, 15% weight loss, severe
DISCUSSION
dehydration, and poor overall appearance at any time during
the experimental period, were euthanized. Thirty-day sur- Herbal products, such as medium chain fatty acids and
vivors were euthanized at the end. Culturing aliquots of essential oils either used as nutritional supplements or
 COCONUT OIL/MONOLAURIN 501

FIG. 1. Depict the microdilution broth culture system (1 mL) employed to test the bactericidal (48 + h) effects of monolaurin on Staphy-
loccoccus aureus BA 42. Serial dilutions of monolaurin and RBD coconut oil were made in nutrient broth and inoculated with exponentially
growing (18 h old) bacteria (6 · 106 cfu). RBD, refined, bleached, deodorized. Color images available online at www.liebertpub.com/jmf

as food preservatives are known to possess antimicrobial
properties.1–6,8–10 For example, we demonstrated earlier that
monolaurin, oregano oil, and some other essential oils
possess antimicrobial and antifungal activity in vitro and
in vivo.5,6,8 Monolaurin is composed of a glycerol unit and a
single lauric acid. Lauric acid is a major component of
virgin coconut oil, but virgin coconut generally contains low
amounts of monolaurin. However, orally administered or
taken as a food additive, some coconut oil is hydrolyzed by
pancreatic lipase into the monoglycerate of lauric acid.11
The RBD coconut oil essentially contains only lauric acid,
because many short and medium chain carbon metabolites,
such as monolaurin are removed.12
However, a therapeutic efficacy has been attributed to RBD
oil and virgin coconut oil based on the hypothesis that lauric
acid can be hydrolyzed in vivo to yield therapeutically ef-
fective monoglycerates (monolaurin) and other medium chain
fatty acid- esters.11,13 Similarly, many coconut oils, used as a
food-flavoring agents, have been postulated to possess a broad
spectrum of antimicrobial activity due to their high content of
lauric acid that can be converted into monolaurin.4 Even
though it is generally accepted that coconut oils containing
high concentrations of lauric acid possess potential antimi-
crobial effects, little actual investigation has been performed
using these natural product to treat superficial or systemic
infections due to bacteria, viruses, or fungi.
Why do many investigators seek effective natural anti-
biotics? There are at least three good reasons. The first is the
possibility of avoiding so-called ‘‘drug resistance’’ whereby
the agent is no longer effective in killing organisms.14,15 In
addition to the possibility of avoiding resistance not un-
common with the use of drugs, many of the synthetic anti-
microbials are expensive (the second reason) and have
shown serious toxic adverse effects (the third reason).16,17
Accordingly, the development of an inexpensive, natural
product free of deleterious health side effects and not prone
to the development of resistance would aid in the preven-
tion, amelioration, and/or cure of various frequent, severe
infectious diseases.
Our study was designed to examine the ability of RBD
coconut oil and virgin coconut oil containing *50% lauric
acid on a fatal form of S. aureus infection in mice. S. aureus
strains (obtained from ATCC) exhibit varying degrees of
sensitivity to antibiotics. The strain BAA 42 appeared to be
sensitive to the antibiotics tested. The strain BAA 14154
was comparatively more resistant to all the antibiotics, in-
cluding methicillin. While the coconut oils produced little
bactericidal effect in vitro, monolaurin showed significant
antibacterial activity, demonstrating bacteriostatic and
bactericidal effects. The concentrations at which mono-
laurin exhibits bactericidal actions on these two strains is
indicative of their biological characteristics. The sensitive
502 MANOHAR ET AL.

FIG. 2. Depict the microdilution broth culture system (1 mL) employed to test the bactericidal (48 + h) effects of RBD coconut oil on
Staphyloccoccus aureus BA 42. Serial dilutions of monolaurin and RBD coconut oil were made in nutrient broth and inoculated with expo-
nentially growing (18 h old) bacteria (6 · 106 cfu). Color images available online at www.liebertpub.com/jmf

and less virulent strain, BAA42 required far less antibiotic

FIG. 3. The number of surviving mice on the different regimens
after 30 days.
and monolaurin. It is important to keep in mind that food
products by nature are not toxic; hence, often require higher
concentrations to bring about the effects exhibited by the
antibiotics (which are require much lower concentrations
but can be quite toxic). Suffice it to say, the above described
features of the natural products could make for good pro-
phylactic agents.
In contrast to concentrated monolaurin, the two coco-
nut preparations did not show any significant effect on
S. aureus. These negative results with the two coconut oils
in the initial in vitro studies were not totally unanticipated.
We expected this, since we had previously postulated that
the coconut oil, dense in lauric acid, needed to be converted
into monolaurin to be effective. Thus, it could only be ef-
fective in vivo where the production of monolaurin could
take place. The virgin coconut oil preparation in vitro
showed some but only minor bactericidal capability con-
sistent with the presence of low levels of monolaurin known
to be present. The lack of any apparent effect from the RBD
coconut oil might be ascribed to the methodology that by
refining, bleaching, and deodorizing removes many fatty
acids that might include monolaurin from the preparation.
What was not fully characterized is whether one can in-
gest enough of precursor lauric acid from the two coconut
preparations to make enough monolaurin to be effective.4 It
is not been established from the literature with certainty that
coconut oils containing lauric acid (50% of the total fatty
 COCONUT OIL/MONOLAURIN
acid present in the coconut) can be metabolized in vivo to
yield sufficient quantities of monolaurin, the glycerol salt of
lauric acid.7 Nevertheless, it seemed possible that coconut
oil could effectively inhibit bacterial growth in vivo when
administered orally. Similar to previous studies,5 vanco-
mycin and monolaurin protected roughly one half the mice
from infectious death (vancomycin 7/12; monolaurin 6/12).
Percentage-wise the combination of the two did even
slightly better (7/10), but larger numbers are needed to de-
cide whether a combination would be a further improve-
ment. The survival patterns with the RBD (0/12) and the
coconut oil (2/12) were essentially similar to that of the
control group (1/12). Based on these findings, we were not
able to show that ingesting large amounts of lauric acid will
produce sufficient monolaurin to be effective.
Perhaps, we did not give the mice enough coconut oil to
be effective. Based on calculations of the amount of lauric
acid found in Mother’s milk, Enig suggested a successful
lauric acid diet for the average human adult would contain
*24 g of lauric acid daily contained in about 3.5 table-
spoons of coconut oil.18 How does this relate to what we
gave the mouse? A tablespoon is roughly 15 mL, therefore, a
reasonable human daily dose calculates to 52.5 mL. Each
mouse weighed *33 g. Comparison on a weight basis
would require that one multiply the mouse dose by roughly
2100 to approximate the weight of a 70 Kg person. Taking
into account surface area,19,20 the equivalency factor for a
mouse (1/12) would make that adjusting ratio closer to 175.
Accordingly, the mouse receiving 0.75 mL daily would take
in a dose equivalent to a human of 131 mL, roughly 2–3
times human daily dose mentioned above (on the basis of
weight alone, more than 20 times). While we believe start-
ing the feeding 1 week in advance is sufficient, still to be
determined is whether the metabolism of mice can be
compared to men in this respect.
While we did not unearth evidence that two different
coconut oils can be helpful to overcome S. aureus infec-
tion in vivo, we corroborated as shown earlier that di-
rect monolaurin ingestion is virtually as effective as
vancomycin.5
AUTHOR DISCLOSURE STATEMENT
No competing financial interests exist.
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