Antimicrobial activity of olive oil

MANUEL BRENES*, EDUARDO MEDINA, CONCEPCIN ROMERO, ANTONIO DE CASTRO

*Corresponding autor

Oil health benefits

Food Biotechnology Department, Instituto de la Grasa (CSIC)

Avda. Padre Garca Tejero 4
41012-Seville, Spain

Anno 18 - No. 4

July/August 2007

AgroFOOD industry hi-tech

INTRODUCTION
As a natural process, foods are contaminated and degraded by an
array of microorganisms, the occurrence of them depending on many
factors such as the type of food, pH, aw, temperature and chemical
preservatives. New products are being introduced onto the market,
especially those designated as organics, and there is an increase in
demand for natural preservatives that could prevent the growth of
foodborne pathogens or to delay the onset of food spoilage (1). Thus,
the growing number of foodborne illness outbreaks caused by some
pathogens is of great concern (2). Food microbiologists have
investigated the antimicrobial properties of many plant molecules
derived from herbs, spices, essential oils and foods (3-6), and in
many occasions this activity has been attributed to phenolic
compounds. Catechins, chlorogenic acid, carvacrol, cinnamaldehyde,
thymol and oleuropein are, among others, the most reported
naturally-occurring phenolic compounds with antimicrobial activity (711). Olive oil and olive leaf extracts have been employed in folk
medicine for centuries, and researchers began to correlate the
antimicrobial activity of these products with phenolic compounds at
the end of the sixties (12). Initial studies were focused on the bitter
compound oleuropein and its effect on the table olive fermentation
(13, 14). Oleuropein, which is the major phenolic compound in olive
fruits, is a bitter glucoside formed by glucose, elenolic acid and the
polyphenol hydroxytyrosol. There has been a controversy for years
about the antimicrobial activity of this substance, although most
researchers found bioactivity (15-17). Its use as a natural
antimicrobial food additive, however, can be limited because of its
strong bitter taste. The antiviral effect of elenolic acid was also
observed in vitro during the sixties and calcium elenolate was
proposed as an antiviral drug (18). Unfortunately, studies carried out
in vivo failed and its medical use was abandoned. Antimicrobial
compounds in the wastewaters generated during the olive oil
extraction process have also been investigated (19). These solutions
have represented a big environmental problem in many
Mediterranean countries for years because of their content in
phenolic compounds that does not allow their complete microbial
depuration (20). Simple phenols such as hydroxytyrosol, tyrosol,
catechol and others have been considered responsible of the
antimicrobial activity detected in the olive oil mill wastewater as well
as their toxicity for plant germination (21). In consequence, many
chemical and biological methods have been investigated to eliminate
these substances from the olive oil mill wastewaters (22) in order to
use them for agricultural purposes. In the case of olive oil, very few
studies have been carried out until now dealing with its antimicrobial
activity. Some researchers detected bactericidal activity in some

ABSTRACT
Among the benefits that the consumption of olive oil can exert on
human health, its antimicrobial activity emerges as a promising new
property. Olive oil, in particular virgin olive oils with a high content in
certain phenolic compounds, can inhibit the growth of pathogenic
bacteria, this activity being higher than that reported for foods such as
tea, coffee, wine and others. Thus, its use in many foods could
contribute to a decrease in foodborne illness outbreaks. Also, olive oil
polyphenols, in particular the dialdehydic form of decarboxymethyl
oleuropein and ligstroside aglycons possess a strong bactericidal
activity in vitro against Helicobacter pylori, which opens up the possibility
of considering virgin olive oil as a chemopreventive agent for peptic
ulcers or gastric cancer.

volatile compounds of the oil (aliphatic aldehydes) (23), and others

attributed this effect to simple phenols (24). However, simple phenols
such as hydroxytyrosol and tyrosol are not the main phenolic
compounds in virgin olive oil (25).
OLIVE OIL POLYPHENOLS
Despite various reports on phenolic compounds in olive oil, there is
still some confusion about their presence and content in the oils.
Oleuropein, the major phenolic compound in fruits, is present in a
very low amount in olive oils and it was not even detected in many
samples (26). The main phenyl alcohols in virgin olive oil are
hydroxytyrosol and tyrosol, which were considered the most important
polyphenols in olive oil up to the nineties when Montedoro et al. (25)
disclosed that their concentration in fresh virgin olive oils is low.
However, a hydrolysis reaction of the secoiridoid aglycons during
olive oil storage occurs and the oil is progressively enriched in these
two phenyl alcohols, the rate of this reaction depending on the quality
of the oil (27). By contrast, the prevalent phenolic compounds in virgin
olive oils are the secoiridoid derivatives such as the oleuropein and
ligstroside aglycons, and the dialdehydic form of decarboxymethyl
oleuropein and ligstroside aglycons (25, 26). These secoiridoids arise
from the glucosides oleuropein and ligstroside present in the olive
fruit which are hydrolyzed by endogenous -glucosidase during
crushing and, thus, are transferred to the oil phase. The lignans, 1acetoxypinoresinol and pinoresinol, and hydroxytyrosol acetate have
also been detected in most olive oils (28), as well as the flavones
luteolin and apigenin. Other minor polyphenols identified in olive oils
were hydroxytyrosol glycol, catechol, tyrosol acetate, p-coumaric,
vanillic, cinnamic and ferulic acids and vanillin. A broad range of
polyphenol concentration in olive oils has been reported for years,
which is due to the high number of factors that influence this
parameter: variety, irrigation, degree of maturation, extraction
process, storage conditions and others. The polyphenols
concentration of Spanish
virgin olive oils ranges
from about 300-500 mg/kg
(29), which is similar to
the range reported for
Italian and Greek oils (25,
30). Of course, there are
commercial olive oils in
Table 1. Phenolic compounds in
Spanish virgin olive oils
the market with higher
and lower amounts. Table
1 shows the range concentration for each individual phenolic
compound in Spanish virgin olive oils. Despite variability, it can be
deduced that the secoiridoid oleuropein derivatives are the main
polyphenols in oils, followed by the secoiridoid ligstroside derivatives
and the lignans. Other minor compounds may account for 10-160
mg/kg of oil.
IN VITRO ANTIMICROBIAL ACTIVITY OF OLIVE OIL
The bactericidal activity of many different types of olive oils against
several microorganisms has been studied in vitro (31). The first
relevant finding of this work was that all types of olive oils killed
bacteria and this effect was not observed for other edible vegetable
oils (corn, sunflower, soybean, rapeseed and cotton). Moreover, this
activity was higher in virgin olive oils, followed by olive oils and
pomace olive oils, which is in accordance with their decreasing

Some researchers have attributed this bioactivity to the simple phenols

hydroxytyrosol and tyrosol (24) and others speculated about other
different polyphenols (34).
We have demonstrated for the first time that the most bactericidal
polyphenols of olive oil are the dialdehydic form of decarboxymethyl
oleuropein and ligstroside aglycons (Figure 1), in particular the latter
compound. We isolated all olive oil polyphenols by HPLC and tested
them against L. monocytogenes and H. pylori (31, 37).
In both cases, the dialdehydic form of decarboxymethyl ligstroside
aglycon showed the strongest bactericidal activity followed by the
oleuropein derivative. No significant effect was observed for
hydroxytyrosol and tyrosol and the oleuropein and ligstroside
aglycons. It seems therefore that the oleosidic part of the active
molecule is very important to exert the killer action.
ANTIMICROBIAL ACTIVITY OF OLIVE OIL IN FOOD
PRODUCTS AND COMPARISON WITH OTHER
FOODSTUFFS
It is well-demonstrated that olives and their derived products,
especially olive oil, possess compounds with antimicrobial activity but
this bioactivity has been reported for many other foodstuffs as well.
The survival of pathogens in common beverages depends on many
factors such as pH, content in organic acids, ethanol, additives and
natural antimicrobials (41-44). Among the latter substances, the
phenolic compounds have been considered the main antimicrobials in
tea, coffee, aromatic plants and many other foodstuffs (10, 45-47).
Given the above findings on the antimicrobial activity of olive oil, we
ran some experiments to compare the survival of pathogenic bacteria
in olive oil extracts and several common beverages (40). S. aureus, L.
monocytogenes, S. enterica, E. coli, S. sonnei and Yersinia sp. were
inoculated in liquid products and counted after 5 minutes of contact.
All tested pathogens survived in peach, pineapple and orange juices,
cow's milk, yogurt drink, coffee extracts, beer with and without alcohol
and Coca-cola. By contrast, red and white wine, green and black tea,
vinegar and olive oil extracts showed bactericidal activity against the
foodborne pathogens tested (Figure 2).

7
AgroFOOD industry hi-tech

Figure 2. Antimicrobial activity of several food products

Oil health benefits

content in phenolic compounds. All bacteria were sensitive to olive oil

polyphenols, the effect being in general higher against Gram-positive
than Gram-negative bacteria. In contrast, the yeast Candida albicans
survived after treatment with olive oil, which is surprising since a
mixture of honey, olive oil and beeswax has been recently proposed to
treat the diaper dermatitis produced partly by this yeast (32), and an
olive leaf extract was able to inhibit the growth of C. albicans after 1
day of contact (33). Moreover, olive oils exerted bactericidal activity in
vitro against the enteric microorganisms of the intestine Escherichia
coli and Clostridium perfringens but also against the beneficial bacteria
Lactobacillus acidophilus and Bifidobacterium bifidum.
These findings have been confirmed by Italian researchers (34).
Hence, it seems that the consumption of olive oil could influence the
growth of the intestinal microbiota, although it would depend on the
amount of oil ingested and the absorption of polyphenols before they
reach the colon (35).
In our work, we also observed a bactericidal effect with all types of
olive oils against the cariogenic bacteria Streptococcus mutants (31)
and it has also been reported by other researchers that an olive oil
formulation dentifrice can decrease both bacterial growth and
adhesion (36). The most promising results of our studies were the
strong bactericidal activity exerted by olive oil polyphenols against
foodborne pathogens.
This activity was found against strains of Staphylococcus aureus,
Listeria monocytogenes, Salmonella enterica subs.enterica, Yersinia
sp and Shigella sonnei, and these findings open up the possibility of
using olive oil as a food preservative to prevent the growth of
foodborne pathogens or to delay the onset of food spoilage (31).
The antimicrobial action of olive oil against the Gram-negative bacteria
Helicobacter pylori (37) was also studied in more detail because this
microorganism is responsible for most peptic ulcers and gastric
cancers (38). Spanish virgin olive oils showed bactericidal activity
against eight strains of H. pylori, five of them isolated from patients.
Likewise, it was confirmed that olive oil polyphenols resist gastric
conditions for hours without any significant transformation. Studies in
vivo are in progress to confirm these relevant findings and to propose
olive oil as a chemopreventive agent for peptic ulcer or gastric cancer.
One question arises from all these results: which are the compounds
responsible of this strong antimicrobial activity?

July/August 2007

Figure 1. Chemical structure of the main antimicrobial

polyphenols in olive oil

Among them, vinegar showed the strongest bactericidal effect,

followed by the aqueous extract of virgin olive oil, wines and olive oil
and tea extracts. Olive oil has been used to preserve foods for
centuries but the studies to identify the bactericidal compounds are
scarce. Radford et al. (39) observed a faster death rate of S. enterica
in mayonnaise made with virgin olive oil than in that prepared with

Anno 18 - No. 4

Oil health benefits

sunflower oil, and they attributed this effect to the high acidity as well
as simple phenols in olive oil. We have confirmed the bactericidal
action of virgin olive oil in egg and milk mayonnaises inoculated with
S. enterica and L. monocytogenes respectively (40). The counts of
pathogen microorganims were reduced by approximately 3 log cfu/ml
after 30 min. Lettuce was also seasoned with different types of oils,
lemon juice and vinegar, and inoculated with L. monocytogenes.
Slight reductions of the pathogen on lettuce seasoned with sunflower
oil and vinegar or lemon juice alone were observed. By contrast, L.
monocytogenes cells were killed after 30 min when virgin olive oil was
used, regardless of the lemon juice or vinegar added.
In light of these results, it can be said that olive oil, especially olive
oil polyphenols, must be considered in the future as a protective food
against foodborne pathogens.
Olive oil is consumed directly on toast and in fresh salads, but it is
also employed in commercially processed foods (e. g. tuna, tomato,
mayonnaise) and in many homemade dishes.
In consequence, olive oil could be a hurdle component in certain
processed foods and exert a protective effect against foodborne
pathogens when contaminated foods are ingested.
ACKNOWLEDGEMENTS
This study was supported by a grant from the Spanish Government
(CICYT) and the European Union FEDER Funds (Project AGL-200300826).

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