Advances in Cytology & Pathology

Mini Review Open Access

Antibacterial activities of rosemary essential oils and

their components against pathogenic bacteria
Abstract Volume 3 Issue 4 - 2018

There are many known essential oils, of which only 300 are commercially important for
Jasmina Stojiljkovic,1 Metodija Trajchev,2
different types of industry. Antimicrobials are used in food for two main reasons: to control
natural spoilage processes and to prevent/control growth of micro-organisms, including
Dimitar Nakov,2 Milena Petrovska3
1
Food Technology, College of Applied Studies, Republic of Serbia
pathogenic microorganisms. The aim of this review was to investigate antibacterial effects 2
Faculty of Agricultural Science and Food, University St. Cyril
of rosemary essential oils (EOs) on some Gram-positive and Gram-negative bacteria: and Methodius, Republic of Macedonia
Staphylococcus aureus, Bacillus subtilis, Escherichia coli, Proteus vulgaris, Pseudomonas 3
Faculty of Medicine, University St. Cyril and Methodius,
aeruginosa. Listeria monocytogenes, Salmonella enteritidis, Salmonella typhimurium. Republic of Macedonia
Rosmarinus officinalis essential oil is very important for its medicinal uses and its powerful
antibacterial, cytotoxic, anti-mutagenic, antioxidant, anti-phlogistic and chemopreventive Correspondence: Jasmina Stojiljkovic, MSc of Technological
properties. The antimicrobial and other biological activities of rosemary essential oil are Microbiology, Coordinator of the department Food Technology,
directly correlated with the presence of bioactive volatile components. To this end, one College of Applied Studies, Filipa Filipovica 20,Vranje, Republic
possibility has been the use of rosemary essential oils (EOs) and the compounds found of Serbia, Tel +3816 3108 6667, Fax +3811 7404 661,
there in as alternative antimicrobial food preservatives. The future will see investigation Email [email protected]
of food applications of the naturally occurring antimicrobials, especially the effectiveness
of rosemary EOs, individually and in combination with other parts of plant extract, other Received: February 05, 2018 | Published: September 18, 2018
effective EOs and other food-processing techniques.

Keywords: antibacterial effect, essential oil, rosemary, pathogenic bacteria

Abbreviations: EOLs, essential oils; ATP, adenosine three Discussion

phosphate; MIC, minimum inhibitory concentrations; IZ, inhibition
zone; ATCC, american type culture collection; NCTC, national Mechanisms of action of the EOs on pathogenic
collection of type cultures, a culture collection of public health bacteria
england Some studies have demonstrated that whole EOs usually
have higher antibacterial activity than the mixtures of their major
Introduction components, suggesting that the minor components are critical to
Plants producing essential oils belong to various genera in about the synergistic activity, though antagonistic and additive effects have
60 families, including Alliaceae, Apiaceae, Asteraceae, Lamiaceae, also been observed.9,10 EOs derived from spices and plants have
Myrtaceae, Poaceae, and Rutaceae.1 Plant essential oils are a antimicrobial activity against Listeria monocytogenes, Salmonella
potentially useful source of antimicrobial compounds.2 Aromatic typhimurium, Escherichia coli O157:H7, Shigella dysenteriae,
plants and their essential oils (EOs) have been used since ancient times Bacillus cereus and Staphylococcus aureus at levels between 0.2 and
in food, agriculture, medicine, cosmetic applications, as condiments 10μL/mL.11 Determination of the antimicrobial activity of 17 essential
and spices, in therapeutic uses, as antimicrobials, as flavoring agents, oils against Escherichia coli O157:H7 and Salmonella enterica in
and in storage as insecticidal agents.3 The use of plant essential oils has apple juices indicated that the reduction in the number of bacteria
been the new focus of agricultural and food industries as an alternative can reach 50%.12 Considering the large number of different groups
to synthetic food additives for inactivation of microorganisms such as of chemical compounds present in EOs, it is most likely that their
pathogenic and spoilage bacteria that cause food safety and public antibacterial activity is not attributable to one specific mechanism but
health-related issues during transportation, storage, shelf-life, and that there are several targets in the cell. An important characteristic
packaging.4 Antimicrobials are used in food for two main reasons: of EOs and their components is their hydrophobicity, which enables
to control natural spoilage processes (food preservation), and to them to partition in the lipids of the bacterial cell membrane and
prevent/control growth of microorganisms, including pathogenic mitochondria, disturbing the structures and rendering them more
microorganisms (food safety).5 Natural antibacterial essential oils permeable. Leakage of ions and other cell contents can then occur
have gained an increased interest and are considered as safe and eco- and causes the death of bacterial cell. Generally, the EOs possessing
friendly alternative to control foodborne bacteria and other pathogenic the strongest antibacterial properties against food borne pathogens
microorganisms, notably drugs resistant ones.6,7 In food processing, it contain a high percentage of phenolic compounds such as carvacrol,
is important that proper measures are taken to ensure the safety and eugenol and thymol.11 The mechanisms of action of the EOs include
stability of products for the duration of their shelf life.8 the degradation of the cell wall, damaging the cytoplasmic membrane,

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© 2018 Stojiljkovic et al. This is an open access article distributed under the terms of the Creative Commons Attribution License,
which permits unrestricted use, distribution, and build upon your work non-commercially.
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cytoplasm coagulation, damaging the membrane proteins, increased which suggested a synergic interaction. The incorporation of oregano
permeability leading to leakage of the cell contents, reducing the and rosemary alone (MIC) or combined at different subinhibitory
proton motive force, reducing the intracellular ATP (Adenosine concentrations in vegetable broth resulted in a decrease in the viable
Three Phosphate) pool via decreased ATP synthesis and augmented cell counts of all test strains over 24h. After a 10-min treatment, the
hydrolysis that is separate from the increased membrane permeability oregano and rosemary, either alone or in combination at different sub
and reducing the membrane potential via increased membrane inhibitory concentrations, caused similar decrease (p>0.05) in the
permeability. The cell wall of Gram-negative bacteria is more resistant counts of all assessed groups (or family) of spoilage microorganisms.
to the activity of EOs and their components. The Gram-negative cell The decrease in counts of mesophilic bacteria, enterobacteria and
wall does not allow for the entrance of hydrophobic molecules as fungi was always close to 3.0, 2.5 and 3.1 log cycles, respectively.
readily as Gram-positive bacteria; thus, EOs are less able to affect the Overall, the exposure of the EOs for 10 min caused a greater
cell growth of the Gram-negative bacteria.13 Interaction of essential decrease (p<0.05) in spoilage native flora counts in comparison to
oils with microbial cell membranes results in growth inhibition 5 min. The essential oil of rosemary were tested against eight strains
of some Gram-positive and Gram-negative bacteria.14 It has been of bacteria: Staphylococcus aureus (NCTC 6571), Bacillus cereus
reported that Gram-positive bacteria appear to be more susceptible (ATCC 11778), Bacillus subtilis (NCTC 10400), Bacillus pumilis
to the antibacterial properties of essential oil compounds than Gram- (wild type), Pseudomonas aeruginosa (NCTC 1662), Salmonella
negative bacteria. This is expected, as Gram-negative bacteria have poona (NCTC 4840), Escherichia coli (ATCC 8739) and amphicilin
an outer layer surrounding their cell wall, limiting the access of resistant Escherichia coli (NCTC 10418). The results from the disc
hydrophobic compounds. Gram-negative bacteria are usually more diffusion assay followed by modified resazurin assay indicated that
resistant to the plant-origin antimicrobials and even show no effect, the tested essential oil showed higher antibacterial activity against
compared to Gram-positive bacteria.15,16 Gram-positive bacteria (IZ 18.0- 24.2; MIC 0.20-0.48mg mL-1)
than against Gram-negative bacteria (IZ 12.8-17.5; MIC 1.16-1.72
Essential oil of rosemary mg mL-1).25 Rosmarinus officinalis Linn. (rosemary) hydroalcoholic
Rosemary (Rosmarinus officinalis L.) is of considerable importance extract was assayed against Streptococcus mitis, Streptococcus
in term of its great an important medicinal and aromatic value. This sanguinis, Streptococcus mutans, Streptococcus sobrinus and
plant belongs to Lamiaceae family. Rosemary is a perennial evergreen Lactobacillus casei standard strains, and its antimicrobial activity
herb with fragrant needlelike leaves.17 Rosemary herbs have been was proven in all tests, except against S. mitis.26 The aim of the
widely used in the traditional medicine and cosmetics. They are study of,27 was to evaluate antibacterial properties of commercial
also used as flavouring agents in foods.18 Rosmarinus officinalis essential oils (rosemary, caraway and fennel) to reduce the number
essential oil is also important for its medicinal uses and its powerful of Staphylococcus aureus and Escherichia coli. The result of
antibacterial, cytotoxic, antimutagenic, antioxidant, antiphlogistic and experiments showed that essential oils contained in microbiological
chemopreventive properties.19 The major components determined in media significantly reduced the number of S. aureus and E. coli cells.
R. officinalis essential oil were 1,8-cineol (38.5%), camphor (17.1%), The best antibacterial properties possessed caraway oil: 1mg/g for S.
α-pinene (12.3%), limonene (6.23%), camphene (6.00%) and linalool aureus and 10mg/g for E. coli, weaker rosemary (5mg/g) and fennel
(5.70%).20 The chemical analysis of rosemary essential oil by GC has (20mg/g) oils. The combination of clove and rosemary essential oils
identified 11 compounds which they make 78.25% rosemary essential produced an additive effect against the Gram-positive and Gram-
oil. Major components of the essential oil include alpha-pinene negative bacteria, namely Staphylococcus aureus, Staphylococcus
(23.93%), camphen (8.7%), camphor (10.97%), verbenon (15.44%), epidermidis, Bacillus subtilis, Escherichia coli, Proteus vulgaris and
p-cymene (7.48%) and 3-Octanone (5.63%).21 Pseudomonas aeruginosa.28 Essential oils Origanum vulgare and
Rosmarinus officinalis combined at sub-inhibitory concentrations
The antimicrobial activity of rosemary oil were effective in inhibiting the growth and survival of pathogenic
and spoilage microorganisms associated to minimally processed
In the Italian territory, Abruzzo is one of the central area regions
vegetables, although the underlying mode of action remains to be
characterized by a multitude ofenvironments and microclimates and
explored in the future. Sensory evaluation suggested that application
with the richest flora of Italy and the Mediterranean basin.22 In the
of the essential oils in mixture at sub-inhibitory concentrations as
Abruzzo territory, different plant species are cultivated and exploited
sanitizer in vegetables would be acceptable to consumers, mainly when
for their therapeutic and alimentary properties;23 among them,
regarded a more extended storage time.29 The inhibitory effects of the
Rosmarinus officinalis, Origanum vulgare, Salvia officinalis, Mentha
hydrosols of thyme, black cumin, sage, rosemary, and bay leaf against
piperita, Allium sativum, Foeniculum vulgare, Satureja montana,
S. typhimurium and Escherichia coli O157:H7 were shown in carrots
Thymus vulgaris, and Coriandrum sativum.
and apples. Thyme hydrosol had the highest antibacterial effect on
The study of24 assessed the effect of the combined application both Salmonella typhimurium and Escherichia coli O157:H7 counts.30
of essential oils (EOs) from Origanum vulgare L. - oregano and Among Gram-negative, Salmonella spp. was inhibited mostly by EOs
Rosmarinus officinalis L. - rosemary, alone or in combination at of thyme (500ppm), rosemary (5000ppm) and garlic (20000ppm), E.
subinhibitory concentrations, against three pathogenic bacteria that coli was mostly inhibited by thyme (250ppm), cinnamon (2500ppm)
are associated with fresh leafy vegetables: Listeria monocytogenes and cumin (2500ppm).31
(L. monocytogenes), Escherichia coli (E. coli) and Salmonella
enterica Serovar Enteritidis (S. Enteritidis). The minimum inhibitory Conclusion
concentration MIC of oregano was 0.6mL/mL against the test strains Natural antimicrobial extracts from herbs, spices, and plants can
either in single and mixed inoculum. The MIC of rosemary was 5mL/ be used in food industry to prevent growth of foodborne pathogens
mL against L. monocytogenes and E. coli and 10mL/mL against S. and food spoilage microorganisms and to enhance the shelf life
Enteritidis in single inocula, whereas it was 10mL/mL against the and stability. Due to the antioxidant and antimicrobial effects of
mixed inoculum. The fractional inhibitory concentration index of natural ingredients, they can be a good alternative to classical food
the combined EOs was 0.5 against the mixed bacterial inoculum,

Citation: Stojiljkovic J, Trajchev M, Nakov D, et al. Antibacterial activities of rosemary essential oils and their components against pathogenic bacteria. Adv Cytol
Pathol. 2018;3(4):93‒96. DOI: 10.15406/acp.2018.03.00060
 Copyright:
Antibacterial activities of rosemary essential oils and their components against pathogenic bacteria ©2018 Stojiljkovic et al. 95

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Citation: Stojiljkovic J, Trajchev M, Nakov D, et al. Antibacterial activities of rosemary essential oils and their components against pathogenic bacteria. Adv Cytol
Pathol. 2018;3(4):93‒96. DOI: 10.15406/acp.2018.03.00060
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Citation: Stojiljkovic J, Trajchev M, Nakov D, et al. Antibacterial activities of rosemary essential oils and their components against pathogenic bacteria. Adv Cytol
Pathol. 2018;3(4):93‒96. DOI: 10.15406/acp.2018.03.00060