Journal of Ethnopharmacology 265 (2021) 113318

Contents lists available at ScienceDirect

Journal of Ethnopharmacology
journal homepage: www.elsevier.com/locate/jethpharm

Traditional use, phytochemistry, toxicology, and pharmacology of

Origanum majorana L.
Abdelhakim Bouyahya a, *, Imane Chamkhi b, Taoufiq Benali c, Fatima-Ezzahrae Guaouguaou d,
Abdelaali Balahbib e, Nasreddine El Omari f, Douae Taha g, Omar Belmehdi h, Zengin Ghokhan i,
Naoual El Menyiy j
a
Laboratory of Human Pathologies Biology, Department of Biology, Faculty of Sciences, And Genomic Center of Human Pathologies, Faculty of Medicine and Pharmacy,
Mohammed V University in Rabat, Morocco
b
Microbiology and Molecular Biology Team, Center of Plant and Microbial Biotechnology, Biodiversity and Environment, Faculty of Sciences, Mohammed V University in
Rabat, Morocco
c
Laboratory of Natural Resources and Environment, Polydisciplinary Faculty of Taza, SidiMohamed Ben Abdellah University of Fez, B.P.: 1223, Taza-Gare, Taza,
Morocco
d
Mohammed V University in Rabat, LPCMIO, Materials Science Center (MSC), Ecole Normale Supérieure, Rabat, Morocco
e
Laboratory of Zoology and General Biology, Faculty of Sciences, Mohammed V University in Rabat, Rabat, Morocco
f
Laboratory of Histology, Embryology, and Cytogenetic, Faculty of Medicine and Pharmacy, Mohammed V University in Rabat, Morocco
g
Laboratoire de Spectroscopie, Modélisation Moléculaire, Matériaux, Nanomatériaux, Eau et Environnement, CERNE2D, Faculté des Sciences, Université Mohammed V,
Rabat, Morocco
h
Biology and Health Laboratory, Department of Biology, Faculty of Science, Abdelmalek Essaadi University, Tetouan, Morocco
i
Biochemistry and Physiology Laboratory, Department of Biology, Faculty of Science, Selcuk University, Campus, Konya, Turkey
j
Laboratory of Physiology, Pharmacology & Environmental Health, Faculty of Science, University Sidi Mohamed Ben Abdellah, Fez, Morocco

A R T I C L E I N F O A B S T R A C T

Keywords: Ethnopharmacological relevance: Origanum majorana L., is an aromatic and medicinal plant distributed in different
Origanum majorana parts of Mediterranean countries. This species is widely used in traditional medicine for the treatment of many
Traditional use diseases such as allergies, hypertension, respiratory infections, diabetes, stomach pain, and intestinal
Pharmacological actions
antispasmodic.
Bioactive compounds
Aim of the review: This work reports previous studies on O. majorana concerning its taxonomy, botanical
Mechanism insights
description, geographical distribution, traditional use, bioactive compounds, toxicology, and biological effects.
Materials and methods: Different scientific data bases such as Web of Science, Scopus, Wiley Online, SciFinder,
Google Scholar, PubMed, ScienceDirect, and SpringerLink were consulted to collect data about O. majorana. The
presented data emphasis bioactive compounds, traditional uses, toxicological investigations, and biological ac­
tivities of O. majorana.
Results: The findings of this work marked an important correlation between the traditional use of O. majorana as
an anti-allergic, antihypertensive, anti-diabetic agent, and its biological effects. Indeed, pharmacological in­
vestigations showed that essential oils and extracts from O. majorana exhibit different biological properties,
particularly; antibacterial, antifungal, antioxidant, antiparasitic, antidiabetic, anticancer, nephrotoxicity pro­
tective, anti-inflammatory, analgesic and anti-pyretic, hepatoprotective, and antimutagenic effects. Toxicological
evaluation confirmed the safety and innocuity of this species and supported its medicinal uses. Several bioactive
compounds belonging to different chemical family such as terpenoids, flavonoids, and phenolic acids were also
identified in O. majorana.
Conclusions: The results suggest that the pharmacological properties of O. majorana confirm its traditional uses.
Indeed, O. majorana essential oils showed remarkable antimicrobial, antioxidant, anticancer, anti-inflammatory,
antimutagenic, nephroprotective, and hepatoprotective activities. However, further investigations regarding the
evaluation of molecular mechanisms of identified compounds against human cancer cell lines, inflammatory

* Corresponding author.
E-mail addresses: boyahyaa-90@hotmail.fr (A. Bouyahya), chamkhi.imane@gmail.com (I. Chamkhi), benali.taoufiq@gmail.com (T. Benali), f.guaouguaou@
gmail.com (F.-E. Guaouguaou), balahbib.abdo@gmail.com (A. Balahbib), nasrelomari@gmail.com (N. El Omari), douae.taha02@gmail.com (D. Taha),
belmehdiomar@hotmail.fr (O. Belmehdi), biyologzengin@gmail.com (Z. Ghokhan), Nawal.ELMENYIY@usmba.ac.ma (N. El Menyiy).

https://doi.org/10.1016/j.jep.2020.113318
Received 1 May 2020; Received in revised form 22 July 2020; Accepted 22 August 2020
Available online 31 August 2020
0378-8741/© 2020 Elsevier B.V. All rights reserved.
A. Bouyahya et al. Journal of Ethnopharmacology 265 (2021) 113318

process, and microbial infections are needed to validate pharmacodynamic targets. The toxicological investi­
gation of O. Majorana confirmed its safety and therefore encouraged pharmacokinetic evaluation tests to validate
its bioavailability.

Abbreviations MCP-1 Monocyte Chemoattractant Protein 1

MIC Minimum Inhibitory Concentration
AGEs Advanced Glycation End products NF-kB Nuclear Factor-kappa B
ATCC American Type Culture Collection NLRP3 NOD-like Receptor Family, Pyrin Domain Containing 3
BHA Butylated Hydroxy-Anisole NOX4 NADPH Oxidase 4
BHT Butylated Hydroxy-Toluene OMEO Origanum majorana essential oils
DPPH 2,2-Diphenyl-1-picrylhydrazyl radical PEPCK Phospho-Enol-Pyruvate Carboxykinase
FRAP Ferric Reducing Antioxidant Power RANTES Regulated upon Activation, Normal T Cell Expressed and
G6pase Glucose-6-Phosphatase Presumably Secreted
G-CSF Granulocyte-Colony Stimulating Factor ROS Reactive Oxygen Species
GM-CSF Granulocyte-Macrophage Colony-Stimulating Factor STAT1 Signal Transducer and Activator of Transcription 1
HDL High Density Lipoprotein STZ Streptozotocin-Induced Diabetic
IC50 Half Maximal Inhibitory Concentration TNF-α Tumor Necrosis Factor
LD5 Lethal Dose VEGF Vascular Endothelial Growth Factor
LDL Low Density Lipoprotein VLDL Very Low-Density Lipoprotein
LIF Leukemia Inhibitory Factor VSMCs Vascular Smooth Muscle Cells
MBC Minimum Bactericidal Concentration

(Hachi et al., 2015). Moreover, several previous works have demon­

strated other ethnobotanical priorities of O. majorana L. such as the
1. Introduction
studies of El-Hilaly et al. (2003); El Abbouyi et al. (2014); Ouelbani et al.
(2016); Alaoui and Laaribya (2017), and Zougagh et al. (2019).
Origanum majorana L. (Known as Sahtar or Zaatar in traditional
Origanum majorana L. is rich in phytochemicals such as thymol,
medicine) currently named sweet marjoram, is a medicinal plant of the
carvacrol, tannins, hydroquinone, arbutin, methyl arbutin, vitexin, ori­
Lamiaceae family, a perennial herb of Origanum genus (Prerna and
entin, thymonin, triacontan, sitosterol, cis-sabinene hydrate, limonene,
Vasudeva, 2015), with a self-supporting growth habit, it is a photoau­
terpinene, camphene and flavonoids like diosmetin, luteolin, and api­
totroph (Ietswaart, 1980). This plant is distributed around the Medi­
genin (Jelali et al., 2011; Guerra-Boone et al., 2015; Sefeer and Elu­
terranean regions, in particular, Morocco, Algeria, Egypt, Spain, and
malai, 2018; Chaves et al., 2019), which explain its biological
Portugal (Ietswaart, 1980).
properties.
In folk Moroccan medicine, marjoram is used as anti-cooling (Bel­
The pharmacological investigations of the methanol extracts and the
lakhdar et al., 1991) against allergies, fever, flu, hypertension (Tahraoui
essential oils of O. majorana have shown interesting biological activities
et al., 2007; Benali et al., 2017), antipyretic (Bellakhdar et al., 1991),
(Fig. 1). In detail, an antibacterial property against different pathogenic
respiratory infections (Ennacerie et al., 2017), antidiabetic (Benkhnigue
bacteria such as Bacillus subtilis, Enterococcus faecalis, Escherichia coli,
et al., 2014; El Hafian et al., 2014), menstrual pain, cold in the uterus,
Klebsiella pneumonia, Salmonella choleraensius, Serratia sp., etc. (Busatta
stomach pain, cough (Bouayyadi et al., 2015), rheumatism, headache,
et al., 2008; Hajlaoui et al., 2016; Amor et al., 2019). The antibacterial
insomnia (Abouri et al., 2012), and used as intestinal antispasmodic

Fig. 1. Pharmacological properties of O. majorana.

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A. Bouyahya et al. Journal of Ethnopharmacology 265 (2021) 113318

action of OMEO bioactive compounds such as carvacrol and thymol Longitubus, Chilocalyxn, Majoranan, Campanulaticalyx, Elongatispica,
involves several mechanisms including the increase of membrane Origanum, and Prolaticorolla) following Ietswaart (1980) classification.
permeability, the leakage of vital cell contents, and the inhibition Our plant O. majorana L. belongs to Majorana (Miller) section which
quorum sensing (Xu et al., 2008). Moreover, O. majorana exhibited consists of three species (Origanum majorana L., Origanum onites L.,
antifungal effects against pathogenic fungi like Aspergillus niger, Tri­ O. syriacum L. var. syriacum, var. bevanii (Holmes) Ietswaart and var.
choderma viride, Penicillium cyclopium, Phytophthora infestants, Candida sinaicum (Boissier) Ietswaart (1980).
sp., etc. (Vagi et al., 2005; Hajlaoui et al., 2016; Della Pepa et al., 2019;
Thanh et al., 2019). The antiparasitic properties of O. majorana, in 3.2. Botanical description
particular, insecticidal (Barbouche et al., 2001) and larvicidal activities
(El-Akhal et al., 2016; Chaves et al., 2019), have also been reported. Origanum majorana L. is a bushy perennial plant that can reach
Moreover, O. majorana was reported to exhibit antidiabetic activity 30–60 cm in height, and possesses descending multi-branched reddish
(Pimple et al., 2012), nephrotoxicity protective effect (Soliman et al., square stems that spill over to create a mound. Its stems are straight
2016), anti-inflammatory, analgesic, and anti-pyretic actions (Seoudi having weak, hairy, round, and green with red speckles. The leaves of
et al., 2009). The antidiabetic properties of O. majorana phenolic com­ this plant are herbaceous, globose, simple, petiolate, and ovate to
pounds involve the increase of plasma insulin, the stimulation of hepatic oblong-ovate, small (0.5–1.5 cm long and 0.2–0.8 cm wide), often
glycogen synthesis, the increase of glucokinase activity, and the quadrigonus-cylindrical, and the texture is extremely smooth and hairy.
down-regulation of glucose-6-phosphatase (G6Pase) and phosphoenol­ The bracts are different from the leaves, densely imbricate, as long as
pyruvate carboxykinase (PEPCK) (van Son et al., 2011). In addition, the calyces, grey green in color, and arranged opposite to each other on a
hepatoprotective (Mossa et al., 2013), the antimutagenic (Qari, 2008), square stem. Sweet marjoram has small flowers (0.3 cm long and ar­
and the gastrointestinal effects were also reported (Khatab and Elhad­ ranged in burr-like, 1.3 cm long heads), hermaphrodite or female. The
dad, 2015). Moreover, the toxicological investigations have shown that flowers are tubular with two lipped, 1-lipped for one or more upper lips
O. majorana ethanolic extract is quietly safe (Seoudi et al., 2009). In entire or denticulate. It has flattened calyces, white or pale pink flowers
addition, no mortalities were recorded for the ethanolic extract of with grey green bracts that bloom in a spike from June to September.
O. majorana in rats according to the study of Selim et al. (2013). Although, the seeds ripen from August to September. They are oval, dark
This review was designed to explore all the studies about the and brown in color. The roots are sub–cylindrical, longitudinally wrin­
O. majorana L. plant; taxonomy, botanical description, distribution, kled with transverse fissures; 0.2–0.6 mm in diameter. The outer surface
ethnobotanical priorities, all pharmacological investigations of the is dark brown while light brown internally with several long rootlets
different parts of this plant, and we will summarize the list of all (Ietswaart, 1980; Prerna and Vasudeva, 2015).
phytochemical components isolated and identified from the methanol
extracts or from the essential oil of this plant. This article aims to provide 3.3. Geographic distribution
a scientific basis for further studies and development of medicinal agents
from O. majorana. Origanum majorana is commonly known as native to Cyprus, Antalya
(Turkey), distributed in different parts of Mediterranean countries as
2. Research methodology Serbia, Italy, Corsica, southern Spain and Portugal, Morocco, and
Algeria (Ietswaart, 1980). Including, all over the world where it is
The literature on O. majorana botanical description, traditional uses, cultivated in many countries in Europe such as France, US, Asia, in
bioactive compounds, pharmacological effects, and toxicological eval­ different parts of India, Hungary, and United States (Prerna and Vasu­
uations were collected, analyzed and summarized in this review. Sci­ deva, 2015).
entific search engines such as PubMed, ScienceDirect, SpringerLink,
Web of Science, Scopus, Wiley Online, Scifnder, and Google Scholar 3.4. Ethnomedicinal use
were used to collect all published articles about this species. Several
terms were used as keywords such as Origanum majorana, Origanum Origanum majorana is one of the medicinal plants known by their use
majorana essential oils, Origanum majorana extracts, antioxidant effects in traditional medicine (Table 1). The therapeutic application depends
of Origanum majorana, anticancer effects of Origanum majorana, anti­ on the plant parts. In traditional Moroccan medicine, the leaves repre­
parasitic effects of Origanum majorana, the chemical composition of sent the most used part, in particular, as anti-cooling (Bellakhdar et al.,
O. majorana essential oils. All published works on O. majorana in 1991; Benlamdini et al., 2014) and antipyretic (Bellakhdar et al., 1991),
different languages was cited in this study. The identification and ex­ also against allergies, fever, flu, and hypertension (Benali et al., 2017).
amination of the collected manuscripts were based on their titles and In addition, the leaves are used as a decoction or powder in the treat­
abstracts. References lists of the retrieved papers were also examined to ment of respiratory infections (Ennacerie et al., 2017), and blood sugar
identify further relevant papers. Chemical structures were drawn using regulation (Ghourri et al., 2013; Benkhnigue et al., 2014; El Hafian et al.,
ChemDraw Pro 8.0 software. PubChem database was used to check the 2014). O. majorana is used in the case of an irritated throat (El Azzouzi
IUPAC names of the secondary metabolites reported from the plant. and Zidane, 2015) as well as for menstrual pain, cold in the uterus,
stomach pain, and cough (Bouayyadi et al., 2015). Moreover, the plant
3. Results and discussion after infusion has been used for hypertension (Tahraoui et al., 2007).
The leaves of O. majorana mixed with stems have been known to be
3.1. Synonym and taxonomy effective against rheumatism, stomach pain, headache, fever, cough,
and insomnia (Abouri et al., 2012). Thus, the leaves mixed with the
Origanum majorana Linnaeus. (ID: 268884), sweet marjoram as a flowers, under the infusion preparation mode, exert effective effects
common name, formerly known as Majorana hortensis Moench and as a (calming and intestinal antispasmodic) and against certain diseases such
heterotypic synonym known as Origanum dubium Boiss (https://www. as colds, fever, and headaches (Hachi et al., 2015). In another ethno­
ncbi.nlm.nih.gov). Origanum majorana L. is a herbaceous species be­ botanical study, El-Hilaly et al. (2003) showed that the branch of
longs to Lamiaceae family which is one out of 200 genera, tribe Men­ O. majorana prepared by infusion is used to treat the fever, cough, and
theae, Origanum genus and Subgenus Majorana (Chishti et al., 2013). flatulence. Thus, the whole plant has sedative and stomach effects (El
The genus consists of over 44 species, 6 subspecies, 3 botanical varieties, Abbouyi et al., 2014), as well as other actions on oral and digestive
and 18 hybrids (Prerna and Vasudeva, 2015). The genus Origanum has diseases (Alaoui and Laaribya, 2017; Zougagh et al., 2019). The whole
been divided into 10 sections (Amaracus, Anatolicon, Brevifilamentumn plant, without its roots, is used for the treatment of headaches, insomnia,

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A. Bouyahya et al. Journal of Ethnopharmacology 265 (2021) 113318

Table 1
Ethnomedicinal use of O. majorana.
Study of area Used part Mode of Traditional use References
preparation

Eastern High Atlas (Morocco) Leaves Decoction Anti-cooling Benlamdini et al.

(2014)
Guercif (NE of Morocco) Leaves Decoction, Allergy, fever, flu, hypertension Benali et al., 2017
infusion
Kenitra (central Morocco) No reported Infusion Respiratory problems, gastric disorders, headache Salhi et al. (2010)
Talassemtane National Park (Western No reported Infusion Neurological and digestive disorders, mouth disease Rhattas et al. (2016)
Rif of Morocco)
Tata Province (South-eastern Morocco) Stems and Decoction Rheumatism, stomach pains, headache, fever, cough, insomnia Abouri et al. (2012)
Leaves Infusion
Oriental Morocco Leaves Not reported Against chill, antipyretlc Bellakhdar et al.
(1991)
Sidi Kacem (central Morocco) Leaves Decoction Respiratory infection Ennacerie et al.
(2017)
El Jadida city and suburbs (Morocco) Whole plant Infusion Sedative, stomachic El Abbouyi et al.
(2014)
Taounate province (Northern Morocco) Branches Infusion Fever, cough, flatulence El-Hilaly et al.
(2003)
Errachidia province (South-eastern Leaves Infusion Hypertension Tahraoui et al.
Morocco) (2007)
Agadir-Ida-Outanane (Morocco) Leaves Decoction Diabetes El Hafianet al.
(2014)
Al Haouz-Rhamna (Morocco) Leaves Decoction Diabetes Benkhnigue et al.
(2014)
Béni-Mellal (Morocco) Leaves Decoction Sore throat El Azzouzi and
Zidane (2015)
Gharb (Morocco) Leaves Decoction Menstrual pain, cold in the womb, ailments of stomach, cough Bouayyadiet al.
(2015)
Khenifra (Morocco) Leaves, flowers Infusion Calming, intestinal antispasmodic, colds, fevers, headaches Hachi et al. (2015)
Moroccan central plateau Aerial parts Infusion Respiratory infection El Hilah Fatima et al.
(2015)
Rural communes Sehoul and Sidi- Whole plant Not reported Digestive ailments Alaoui and Laaribya
Abderrazak (Maamora-Northern (2017)
Morocco)
Settat (Morocco) Whole plant Infusion Headache, insomnia, respiratory ailments, gastric disorders (spasm, colic Tahri et al. (2012)
without the …)
roots
Tan-Tan (Moroccan Sahara) Leaves Powder Diabetes Ghourri et al. (2013)
Casablanca (Moroccan central) Whole plant Decoction Gumdisease, dental pain Zougagh et al.
(2019)
Town of Escolca (Sardinia, Italy) Leaves Infusion Neuralgia, sedative, stomach pain Loi et al. (2005)
Thessaloniki (North of Greece) Aerial parts Decoction, Hypertension, hypotension, antiemetic, bloating, spasmolytic, whooping Hanlidou et al.
infusion cough, calmative, dizziness, headache, migraine, asthma, common cold, (2004)
dysmenorrhoea, antipyretic
Eastern Region (Libya) Leaves, flowers Not reported Flatulence, metritis, cough, menstruation, loss of appetite, heliananthic El-Mokasabial., 2016
colic, tranquilizing for nerves, hypertension, premenstrual syndrome
Alaşehir; Manisa (Turkey) Leaves Infusion Respiratory tract diseases, flu, asthma Sargın et al. (2013)
Antalya (Turkey) Aerial parts Infusion Sore throat, cold Senkardes and
Tuzlaci (2014)
Mersin and Adana provinces (Turkey) Whole plant Not reported Respiratory tract disorders, circulation system disorders, the intestinal Everest and Ozturk
digestive diseases (2005)
East Anatolia (Turkey) Whole plant Maceration Sedative, diaphoretic Altundag and Ozturk
(2011)
Acipayam –Denizli (Turkey) Aerial parts Infusion Cough Bulut et al. (2017)
Kapıdağpeninsula (Turkey) Leaves, seeds Not reported Stomach aches, atherosclerosis Uysal et al. (2010)
East Anatolia (Turkey) Whole plant Maceration Sedative, diaphoretic, stomach ache Özgökçe and
Özçelik, 2004
Constantine and Mila (North-East of Whole plant Not reported Rheumatism, thyroid, respiratory system diseases, cholesterol, Ouelbani et al.
Algeria) hypoglycemic (2016)
Hodna region (Algeria) Flowers Decoction Arterial hypertension Madani et al. (2012)
Catalonia (Spain) Aerial parts Infusion Hypnotic Bonet et al. (1999)

respiratory ailments, and gastric disorders (Tahri et al., 2012). Then, the In Turkey, the traditional use of O. majorana was reported by several
aerial part of O. majorana prepared by infusion is used against respira­ studies (Table 1). Using the maceration, the whole plant has a sedative
tory infections (El Hilah Fatima et al., 2015). Other ethnobotanical effect, diaphoretic, and an effect against stomach aches (Özgökçe and
studies have shown that O. majorana in infusion form has traditional Özçelik, 2004; Altundag and Ozturk, 2011), as well as for respiratory
effects for respiratory problems, gastric disorders, and headaches (Salhi tract disorders, circulation system disorders, and digestive intestinal
et al., 2010), also for neurological and digestive disorders and every­ diseases (Everest and Ozturk, 2005). The leaves are used after infusion
thing related to mouth diseases (Rhattas et al., 2016). The traditional use against influenza and asthma (Sargın et al., 2013). Furthermore, the
of O. majorana in traditional world pharmacopoeia representing by O. majorana leaves mixed with the seeds are used in the case of stomach
various therapeutic uses according to different parts of the plant are ache as well as antiatherosclerosis (Uysal et al., 2010). The aerial parts
represented in Table 1. of O. majorana are used after infusion against cough (Bulut et al., 2017),

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A. Bouyahya et al. Journal of Ethnopharmacology 265 (2021) 113318

as well as in the case of sore throat and as anti-cooling (Senkardes and α-terpinene (14.28%) (Abdalla and Hendi, 2014b).
Tuzlaci, 2014). The work of Raina and Negi (2012) showed that the O. majorana
In Algeria, the whole plant is used against neurological and digestive essential oil is mainly composed of terpinene-4-ol (31.15%),
disorders, rheumatism, thyroid, diseases of the respiratory system, cis-sabinene hydrate (15.76%), p-cymene (6.83%), sabinene (6.91%),
cholesterol, hypoglycemia, and diseases of the mouth (Ouelbani et al., trans hydrate sabinene (3.86%), and α-terpineol (3.71%). In addition,
2016). Moreover, the O. majorana flowers have an effect on hyperten­ terpinene-4-ol, γ-terpinene, cis-sabinene hydrate, and α-terpineol are
sion arterial (Madani et al., 2012; Sari et al., 2012). In Libya, the flowers the main compounds identified in the essential oil of O. majorana leaves
of O. majorana mixed with the leaves have been used against flatulence, (Jelali et al., 2011). However, the main compounds of OMEO cultivated
cough, menstruation pain, loss of appetite, heliananthic colic, hyper­ in China are terpinene-4-ol (33.0%), caryophyllene oxide (11.9%),
tension, and premenstrual syndrome (El-Mokasabi et al., 2016). p-cymene (6.8%), α-terpineol (6.7%), and spathulenol (6.0%) (Jiang
In European countries, O. majorana has been used to treat different et al., 2011). Chromatographic analysis of OMEO reveals the presence of
illnesses (Table 1). In Italy, the O. majorana leaves in traditional medi­ 27 compounds, namely terpinene-4-ol (36.2%), p-cymene (16.3%), and
cine are considered anti-neuralgia and sedative (Loi et al., 2005). Either γ-terpinene (7.31%) (Khanavi et al., 2010). The main compounds of 20
by the infusion or decoction preparation modes, the aerial part of O. Origanum species are cis-sabinene hydrate and cis-sabinene hydrate
majorana are used in Greece against hypertension, hypotension, bloat­ acetate, representing a total of 68.5% of the essential oil of each species
ing, whooping cough, dizziness, headache, migraine, asthma, colds, and (Novak et al., 2003).
dysmenorrhea and also as antipyretic, analgesic, spasmolytic, and The compounds identified in O. majorana essential oil are repre­
antiemetic (Hanlidou et al., 2004). Furthermore, the whole plant is used sented by cis-sabinene hydrate (cis-thuyanol-4) which reaches 33.3%
in Catalonia (Spain) against hypnosis (Bonet et al., 1999). and terpinene-4-ol with 21.6% (Arnold et al., 1993). The 4-terpineol
compound identified in O. majorana essential oil is the major com­
3.5. Phytochemistry pound with 37%; α-terpineol hydrate and cis- and trans-sabinene made
up 50% of this oil (Komaitis et al., 1992). The work of Nykänen (1986)
The secondary metabolites produced by O. majorana have been the has shown that cis-sabinene hydrate (8–43% of the oil) and 4-terpineol
subject of several studies. These studies have almost all focused on the (21–52% of the oil) are the main compounds of O. majorana essential oil.
aerial parts of this plant. Phytochemical screening of extracts and The essential oil of O. majorana has a very important chemical
essential oils of O. majorana has shown the richness of this plant in polymorphism. The content and the nature of the major compounds vary
phenolic compounds. These are most commonly phenolic acids, flavo­ considerably from one sample to another depending on the origin of the
noids, and terpenoids. Chromatographic analyzes of O. majorana plants. In addition to the essential oil, O. majorana also contains phenol
essential oils allowed identifying around thirty terpenoid compounds acids, flavonoids, sterols, triterpenes, alkaloids, coumarins, tannins, and
(Table 2, Fig. 2). saponins (Hossain et al., 2011; Benhalilou et al., 2019). have identified
The essential oil of O. majorana is mainly composed of carvacrol, three groups of phenolic compounds in O. majorana extracts; phenolic
linalool, thymol, borneol, camphor, terpinen-4-ol, α-pinene, α-thujene, acids group with five compounds: rosmarinic acid, caffeic acid, gallic
p-cymene, terpinene, α-terpineol, sabinene, myrcene, limonene, acid, carnosic acid, and ferulic acid; flavonoids group with eight com­
camphene, terpinolene, verbenene, β-caryophyllene, 1,8-cineole, euca­ pounds: luteolin-7-Oglucoside, apigenin-7-Oglucoside, apigenin, hes­
lyptol, and phellandrene (Hajlaoui et al., 2016; Muqaddas et al., 2016; peretin, luteolin, arbutin, quercetin, and catechin; terpenoids group
Erdogan and Ozkan, 2017; García-Risco et al., 2017; Ben Salha et al., with five compounds: carnosol, limonene, terpinen-4-ol, linalylacetate,
2017; Al-Fatimi, 2018; da Cunha et al., 2018; Baj et al., 2018; Jan et al., and β-caryophyllene (Table 2, Fig. 2).
2018; Partovi et al., 2018; Sefeer and Elumalai, 2018; Abbasi-Maleki This chemical composition is completely distinct from that of the
et al., 2019; Amor et al., 2019; Benhalilou et al., 2019; Chaves et al., extracts studied by several authors (Sellami et al., 2009; Kaiser et al.,
2019; Della Pepa et al., 2019; Khadhri et al., 2019; Makrane et al., 2019; 2013; Taamalli et al., 2015; Makrane et al., 2018; Méabed et al., 2018).
Ragab et al., 2019; Thanh et al., 2019; Waller et al., 2019; Xylia et al., Indeed, the phenolic acids are the main group identified in O. majorana
2019). extracts with a dozen compounds, namely gallic acid, caffeic acid,
Chemical variability is observed in the composition of OMEO dihydroxy phenolic acid, chlorogenic acid, syringic acid, vanillic acid,
extracted by different methods; O. majorana essential oils are rich in p-coumaric acid, ferulic acid, rosmarinic acid,
oxygenated monoterpenes and monoterpene hydrocarbons. While, trans-2-dihydroxycinnamic acid, cinnamic acid, lithospermic acid, and
oxygenated sesquiterpenes have the lowest percentage. Terpinene-4-ol pyrogallol. Moreover, several flavonoids have been identified such as
is the major compound (Ragab et al., 2019). epicatechin, rutin, quercetin-3-rhamnoside, luteolin, coumarin, quer­
The work of Chaves et al. (2019) has shown that pegegone (57.05%) cetin, apigenin, amentoflavone, hesperidin, taxifolin, and isorhamnetin
is the major compound of O. majorana essential oil followed by verbe­ (Table 2, Fig. 3).
none (16.92%), trans-menthone (8.57%), cis-menthone (5.58%), On the other hand, the chemical composition of the methanolic
piperitone (2.83%), 3-octanol%), and isopulegol (1.47%). extract of O. majorana was determined by reverse phase high perfor­
The essential oil of O. majorana aerial parts is rich in terpinoids; mance liquid chromatography. Amentoflavone is the dominant flavo­
whose main component is carvacrol with 52.5%, followed by linalool noid. However, trans 2-hydrocinnamic acid is the main phenolic acid
with 45.4%. This essential oil consists mainly of oxygenated mono­ (Baâtour et al., 2013). Ayari et al. (2013) have shown that the meth­
terpenes (98.2%), while monoterpene hydrocarbons are poorly repre­ anolic extract of different organs of O. majorana studied are rich in
sented (1.7%). Terpinene-4-ol is the main component of O. majorana phenolic acids, flavonoids, and tannins.
essential oil from Kalocsa, Hungary (Erdogan and Ozkan, 2017). The Furthermore, the chemical composition of other O. majorana extracts
main compounds identified in O. majorana essential oils treated with 75 is characterized by the strong dominance of phenolic acids. The results
mM NaCl are sabinene (7.723 μg/g DW) followed by cis-sabinene hy­ obtained made it possible to identify approximately 8 constituents
drate (4.857 μg/g DW), and terpinene 4-ol (2.861 μg/g DW) (Olfa et al., (catechol, cinnamic acid, gallic acid, ascorbic acid, syringic acid, caffeic
2016). Chemical analysis of two fractions of O. majorana essential oil has acid, p-coumaric acid, and trans-ferulic acid) (Table 2, Fig. 4) (Dhull
shown that the colorless fraction is rich in terpinene-4-ol (23.1%) and et al., 2016; Makrane et al., 2019). On the other hand, a study was
thymol (16.3%), however these same compounds are also predominant carried out by Adam and Ahmed (2014) on O. majorana extracts and it
in the yellow fraction, terpinene-4-ol (27.7%) and thymol (24.6%) proved the presence of other secondary metabolites such as sterols,
(Guerra-Boone et al., 2015). The main compounds identified in triterpenes, alkaloids, coumarins, tannins, and saponins. The variations
O. majorana essential oil are 4-terpineol (34.23%) followed by encountered in the chemical composition of the essential oil and extracts

5
A. Bouyahya et al. Journal of Ethnopharmacology 265 (2021) 113318

Table 2
Chemical compounds of O. majorana.
Part used Extracts/essential oils Compounds Compounds References
groups

Aerial Essential oil Terpinoids Terpinene-4-ol, γ-Terpinene, trans-Sabinene hydrate, α-Terpinene, Sabinene, Abbasi-Maleki et al.
parts α-Terpineol, α-Terpinolene, β-Phellandrene, Thujene, α-Pinene, β-Pinene, β-Myrcene, (2019)
α-Phellandrene, r-Cymene, cis-Sabinene hydrate, (E)-p-Menth-2-en-1-ol, trans-
Piperitol, Linalyl acetate, Nerylacetate, trans-Caryophyllene, α-Humulene,
Bicyclogermacrene, Spathulenol, Caryophyllene oxide
Aerial Essential oil Terpinoids Terpinen-4-ol, α-Terpinene, endo-Fenchyl-acetate, Terpineol, α-Pinene, p-Cymene, Amor et al. (2019)
parts iso-Sylvestrene, α-Phellandrene, δ-3-Carene, Limonene, 1,8 Cineole, β-Ocimene, cis-
Sabinene hydrate, β-Pinene, Pulegone, trans-Pinocarveol, cis-Limonene oxide,
dihydro-Linalool, cis-Verbenol, Viridene, (E)-Isocitral, Thymol, Carvacrol, γ-Elemene,
α-Terpinylacetate, Eugenol, Nerylacetate, α-Copaene, Geranylacetate, iso-
Longifolene, (E)-Caryophillene, β-Duprezianene, β-Cedrene, β-Copaene, α-Guaiene,
Aromadendrene, allo-Aromadendrene, Valencene, Caryophyllene oxide, Epiglobulol,
(− )-Spathulenol, β-Atlanthol, Rosifoliol, Cubenol
Aerial Essential oil Terpinoids α-Thujene, α-Pinene, α-Fenchene, Camphene, Verbetene, trans-Isolimonene, δ-2- Della Pepa et al.
parts Carene, α-Terpinene, cis-Sabinene hydrate, Linalool, cis-p-Menth-2-en-1-ol, allo- (2019)
Ocimene, trans-p-Menth-2-en-1-ol, Terpinen-4-ol, α-Terpineol, Thymol, Carvacrol,
δ-Elemene, (E)-Caryophyllene, cis-Muurola-3,5-diene
Aerial Essential oil Terpinoids Sabinene, β-Myrcene, β-Cymene, o-Cymol, Eucalyptol, cis-Linalool oxide, 2-Furanm Khadhri et al.
parts ethanol, Nerol, β-Linalool, β-Terpineol, cis-Terpineol, Thujol, Borneol, p-Menth-1-en- (2019)
4-ol, α-Terpineol, Transpiperitol, p-Menth-2-ene 1,4-epidioxy, Bergamiol, 3,3,5-tri­
methyl cyclohexanol, Caren 2 ol, 2 Undecanone, 1 Methyl 1,2 cyclohexene oxide, p-
Menth-2-ene 1,4 epidioxy, p-Menthane-1,2,3-triol, isothujol, Caryophyllene oxide,
Longipinene epoxide, Ledol, Farnesene, trans-Z-α-Bisabolene Epoxide
Aerial Essential oil Terpinoids γ-Terpinene, trans-Sabinene hydrate, α-Terpinene, Sabinene, cis-Sabinene hydrate, Ragab et al. (2019)
parts α-Terpinolene, β-Phellandrene, cis-p-Menth-2-en-1-ol, α-Thujene, D-Limonene, L-
Linalool
Aerial Aqueous extract Phenolic acids Syringic acid, Caffeic acid, p-coumaric acid, trans-ferulic acid, o-Coumaric acid Makrane et al.
parts (2019)
Aerial Essential oil Terpinoids Alpha.-thujene, (+)-Sabinene, Beta.-Myrcene, Alpha.-phellandrene, (+)-4-Carene, D- Makrane et al.
parts Limonene, Beta.-Phellandrene, Beta.-Cymene, Gamma.-Terpinene, 4-Isopropenyl-1- (2019)
methylbicyclo[3.1.0]hexan-1-ol, Linalool, 5-Isopropyl-2-methylbicyclo[3.1.0]hexan-
2-ol, Trans-4-Isopropyl-1-methyl-2-cyclohexen-1-ol, Cis-4-Isopropyl-1-methyl-2-
cyclohexen-1-ol, (− )-Terpinen-4-ol, Alpha.-Terpineol, Trans-p-Menth-1-en-3-ol,
Bergamol, 2-Isopropenyl-1-methyl-4-(1-methylethylidene)-1-vinylcyclohexane,
Carvacrol, Neryl acetate, Caryophyllene, Germacrene B, Spathulenol, Beta.-Eudesmol
Aerial Depleted aqueous Phenolic acids o-Coumaric acid, Syringic acid, Caffeic acid, Vanillin, p-Coumaric acid Makrane et al.
parts (2018)
Aerial Petroleum ether Phenolic acids Syringic acid, Caffeic acid, Vanillin, trans-Ferulic acid, o-Coumaric acid, Cinnamic Makrane et al.
parts acid (2018)
Flavonoids Rutin, Luteolin, Quercetin, trans-Chalcone
Aerial Dichloromethane Phenolic acids trans-Ferulic acid, o-Coumaric acid, Cinnamic acid Makrane et al.
parts Flavonoids Apigenin, Quercetin, Rutin, Luteolin (2018)
Aerial Ethyl acetate Phenolic acids Vanillic acid, 4-Hydroxybenzoic acid, Syringic acid, Caffeic acid, Vanillin, trans- Makrane et al.
parts Ferulic acid, Cinnamic acid (2018)
Flavonoids Luteolin
Aerial Methanol Phenolic acids Vanillic acid, trans-Ferulic acid, o-Coumaric acid Makrane et al.
parts (2018)
Aerial Aqueous extract Phenolic acids Chlorogenic acid, Syringic acid, Caffeic acid, Vanillin, p-Coumaric acid, Ferulic acid, Méabed et al.
parts o-Coumaric acid, Cinnamic acid (2018)
Flavonoids Rutin, Luteolin
Aerial Essential oil Terpinoids Trans-sabinene hydrate, Sabinene, cis-sabinene hydrate, γ-terpinene, α-terpinyl Al-Fatimi (2018)
parts acetate, α-terpinene, terpinen-4-ol, p-Cymene, α-Thujene, β-Phellandrene, α-Pinene,
Terpinolene
Aerial Essential oil Terpinoids Tujene, α-pinene, camphene, sabinene, β-pinene, β-myrcene, α-phellandrene, Salha et al. (2017)
material α-terpinene, p-cymene, β-phellandrene, β-ocimene, γ-terpinene, α-Terpinolene, trans-
sabinene hydrate, cis-sabinene hydrate, linaloool, trans-p-menth-2-enol, 1-terpineol,
endo-borneol, terpine-4-ol, p-cymen-8-ol, α-terpinol, trans-piperitol, dihydrocarvone,
cis-piperitol, carvone, carvacrol, caryophyllene, aromandendrene, α-humulene,
bicyclo germacrene, spathulenol, caryophyllene oxide, linalyl acetate, bornyl acetate,
α-terpenyl acetate, neryl acetate, geranyl acetate
Aerial Essential oil Terpinoids Carvacrol, Linalool, γ-Terpinene, Cymene, Thymol Erdogan and Ozkan
parts (2017)
Aerial Essential oil Terpinoids Sabinene, 1r-pinene, β-phellandrene, β-pinene, β-myrcene, p-cymene, D-limonene, Ouedrhiri et al.
parts 1,8 cineole, E-terpineol, trans-4-thujanol, trans-2-menthenol, cis-menth-2-en-1-ol, (2016)
(− )-terpinen-4-ol, p-cymen-8-ol, α-terpineol, verbenone, cis-sabinene, hydrate
acetate, trans-sabinene hydrate acetate, caryophyllene, spathulenol, caryophyllene
oxide
Aerial Essential oil Terpinoids Terpinene-4-ol, α-Terpinene, Sabinene, α-Terpineol, Linalool, (Z)-β-Ocimene, (Tahmasebi et al.,
parts Terpinolene, (E)-β-Ocimene, trans-Sabinene hydrate, Bicyclogermacrene, Myrcene, 2016)
trans-Caryophyllene
Aerial Essential oil Terpinoids Thymol, cis-Sabinene hydrate, γ-Terpinene, Terpinen-4-ol, Sabinene, α-Terpinene, Soliman et al.
parts Terpinolene, p-Cymene, Bicyclogermacrene, α-Terpineol, β-Pinene, p-Menth-2-en-1- (2016)
ol
(continued on next page)

6
A. Bouyahya et al. Journal of Ethnopharmacology 265 (2021) 113318

Table 2 (continued )
Part used Extracts/essential oils Compounds Compounds References
groups

Aerial Essential oil Terpinoids trans sabinene hydrate, terpinene 4-ol, trans-p-menth-2-1-ol, cis-sabinene hydrate, Olfa et al. (2016)
parts Sabinene, Bicyclogermacreme, Tricyclene, Linalyl acetate, γ-terpinene, neryl acetate,
β-elemene, α-pinene
Aerial Methanolic extractss Phenolic acids Dihydroxybenzoic acid hexose, Syringic acid, Vanillic acid, p-Coumaric acid, Taamalli et al.
parts Rosmarinic acid (2015)
Flavonoids Caffeoyl-arbutin, Gallocatechin isomer 2, Luteolin-6,8-C-dihexose, Orientin,
Apigenin-O-glucuronide, Luteolin, Taxifolin, Dihydrokaempferide, Eriodictyol,
Sakuranetin, Isorhamnetin
Aerial Petroleum ether extract Other secondary Sterols, Triterpens, Alkaloid, Coumarins, Tannin, Saponine Adam and Ahmed
parts Methanol + water metabolites (2014)
Aerial Essential oil Terpinoids 1,30-Dibromotriacontane, 11-Tricosene, 1,38-Dibromooctatriacontane, 1-Pentacon­ Selim et al. (2013)
parts tanol, O-2-Methylpropyl-hydroxylamine, 2-Piperidinone,N-[4-bromo-N-butyl], 2-
Methyl-tricosane, 9-Cyclohexyl-eicosane, 1,54-Dibromo-tetrapentacontane, 2,4,6-
Tritert-butyl-4-methyl-2,5-cyclohexadien-1-one, 1-Hexyl-2-nitrocyclohexane, 2-
Azido-2,3,3-trimethylbutane, 6-Ethyl-2-methyl-decane
Aerial Methanolic extract Phenolic acids Trans-2-hydrocinnamic acid Baâtour et al.
parts Flavonoids Amentoflavone (2013)
Aerial Essential oil Terpinoids Terpinen-4-ol, cis-Sabinene hydrate, Sabinene, p-Cymene, γ-Terpinene, trans- Raina and Negi
parts Sabinene hydrate, α-Terpineol, β-Caryophyllene, Linalyl acetate, Limonene, (2012)
Bicyclogermacrene, trans-p-Menth-2-en-1-ol
Aerial Methanolic extract Phenolic acids Rosmarinic acid, Caffeic acid, Gallic acid, Carnosic acid Hossain et al.
parts Flavonoids Luteolin-7-Oglucoside, Apigenin-7-Oglucoside (2011)
Terpinoids Carnosol
Aerial Essential oil Terpinoids Trans Sabinene hydrate, Terpinene-4-ol, cis-Sabinen hydrate, δ-Terpinene, Sabinene, Alizadeh et al.
parts α-Terpineol, α-Phellandrene, p-Cymene, β-Caryophyllene, Bicyclogermacrene, (2011)
Myrcene, Terpinolene
Aerial Essential oil Terpinoids Thymol, Carvacrol, p-Cymene, Terpinene-4-ol, γ-Terpinene Khanavi et al.
parts (2010)
Aerial Methanolic extract Phenolic acids Gallic acid, Cafeic acid, Dihydroxyphenolic acid, Chlorogenic acid, Syringic acid, Sellami et al. (2009)
parts Vanillic acid, p-Coumaric acid, Ferulic acid, Rosmarinic acid, trans-2-
Dihydroxycinnamic acid, Cinnamic acid
Flavonoids Epicatechin, Rutin, Quercetin-3-rhamnoside, Luteolin, Coumarin, Quercetin,
Apigenin, Amentoflavone
Aerial Essential oil Terpinoids α-Pinene, α-Thujene, Sabinene, 3-Carene, Myrcene, α-Terpinen, Limonene, 1,8- Sellami et al. (2009)
parts Cineole, trans-2-Hexenal, α-Terpinene, p-Cymene, Terpinolene, trans-Sabinene
hydrate, Linalool, cis-Sabinene hydrate, Linalyl acetate, Bornyl acetate, α-Elemene,
Terpinen-4-ol, β-Caryopyllene, α-Humulene, α-Terpinyl acetate, Myrtenyl acetate,
α-Terpineol, Bicyclogermacrene, β-Selinene, Geranyl acetate, γ-Cadinene, Nerol,
Myrtenol, Geraniol, cis-Carveol, Nonadecane, Eicosane, Methyl eugenol, Spathulenol,
Thymol, Carvacrol
Aerial Essential oil Terpinoids cis-sabinene hydrate, Terpinen-4-ol, linalyl acetate, α-Terpineol, trans Sabinene Tabanca et al.
parts hydrate, Linalool, α-Terpinene, Sabinene, bicyclogermacrene, thymol, terpinolene, (2004)
trans-p-menth-2-en-1-ol
Aerial Essential oil Terpinoids Terpinene-4-ol, Terpinolene, α-Pinene, β-Pinene, α-Terpinene, γ-Terpinene, Novak et al. (2003)
parts α-Terpineol, Camphene, 1,8-Cineol, cis-sabinene-hydrate, Sabinene, β-Caryophyllene
Aerial Essential oil Terpinoids Carvacrol, γ-terpinene, p-cymene, myrcene, α-pinene, α-terpineol, α-terpinene, trans- Baser et al. (1993)
parts sabinene hydrate, trans-dihydrocarvone + methyl carvacrol, thymol, 1,8-cineole,
Limonene, terpinen-4-ol, β-phellandrene
Aerial Essential oil Terpinoids α-Pinene, β-Pinene, Sabinene, δ-3-Carene, Myrcene, a-Phellandrene, a-Terpinene, Komaitis et al.
parts Limonene, β-Phellandrene, cis-β-Ocimene, γ-Terpinene, trans-β-Ocimene, p-Cymene, (1992)
Terpinolene, β-Caryophyllene, 1,2,3,4,4a,Hexahydro-1,6-dimethyi-4-isopropyl-
naphthalene,1-Octen-3-ol, trans-Sabinene hydrate, cis-Sabinene hydrate, Linalool, 3-
Octanol, 1,8-Cineol, 4-Terpinenl, trans-trans-(+)-5-Carano, a-Terpineol, c/s-Piperitol,
trans-Piperitol, Geraniol, Santalol, Benzaldehyde, l-(l,4-Dimethyl-3-cyclohexen-l-yl)-
ethanone, Verbenone, Carvone, Estragoi, Thymol, Carvacrol, Ethyl myristate,
Palmitic acid Ethyl paimitate, Ethyl linoleate, 4-Terlx-nyl acetate, Linalyl acetate,
Bornyl acetate, Geranyl acetate
Aerial Essential oil Terpinoids α-Thujene, α-Pinene, Sabinene, β-Pinene, Myrcene, α-Phellandrene, α-Terpinene, p- Nykänen (1986)
parts Cymene, β-Phellandrene, Limonene, Ocimene, γ-Terpinene, Terpinolene,
β-Caryophyllene, α-Humulene, Germacrene, γ-Cadinene
Leaves Ethanolic extract Phenolic acids Rosmarinic acid, Ferulic acid Benhalilou et al.
Terpinoids Limonene, Terpinen-4-ol, Linalylacetate, β Caryophyllene (2019)
Flavonoids Apigenin, Hesperetin, Luteolin, Arbutin, Quercetin, Catechin
Leaves Essential oil Terpinoids Pulegone, verbenone, trans-p-menthan-2-one, isomenthone, piperitone, 3-octanol, Chaves et al. (2019)
isopulegol, Lavandulol, β-pinene, Limonene, α-pinene, nonen-1-al-(2E)
Leaves Essential oil Terpinoids α-Thujene, α-Pinene, Camphene, Sabinene, β-Pinene, 1-Octen-3-ol, 3-Octanone, Jan et al. (2018)
Myrcene, α-Phellandrene, 3-Carene, α-Terpinen, p-Cymene, Limonene,
β-Phellandrene, (Z)-β-Ocimene, γ-Terpinene, trans-Sabinene hydrate, Borneol,
Terpinen-4-ol, α-Terpineol, trans-Dihydrocarvone, Carvacrol methylether,
Thymoquinone, Thymol, Carvacrol, β-Caryophyllene, α-Humulene, Allo-
Aromadendrene, α-Muurolol, β-Bisabolene, γ-Cadinene, δ-Cadinene, Spathulenol,
Caryophyllene oxide, Epi-α-Muurolol, α-Eudesmol
Leaves Aqueous lyophilized extract Phenolic acids Chlorogenic acid, Gallic acid, Pyrogallol, Resorcinol, p-Coumaric acid, Ferulic acid, Makrane et al.
Cinnamic acid (2018)
Flavonoids Hesperidin, Quercetin, Rutin
(continued on next page)

7
A. Bouyahya et al. Journal of Ethnopharmacology 265 (2021) 113318

Table 2 (continued )
Part used Extracts/essential oils Compounds Compounds References
groups

Leaves Supercritical fluid extractor Terpinoids Eucalyptol, Linalool, Camphor, Borneol, Terpineol, Thymol, Carvacrol, γ-Elemen, García-Risco et al.
α-Terpineol acetate, Neryl acetate, Caryophyllene, Longiborneol, α-Elemen, (2017)
α-Himachalen, γ- Cadinene, Epiglobulol, Globulol, Ent-spathulenol, Caryophyllene
oxide, Ledol, γ- Eudesmol, β- Guaiene, β-Eudesmol
Leaves Essential oil Terpinoids cis-Sabinene hydrate, Terpinen-4-ol, γ-Terpinene, trans-Sabinene hydrate, Sabinene, Calín-Sánchez et al.
(fresh) α-Terpinene, Linalool, Linalyl acetate, α-Thujene, α-Pinene, Camphene, β-Myrcene, (2015)
β-Pinene, 2-Hexen-1-ol acetate, 3-Carene, p-Cymene, Limonene, cis-β-Ocimene,
Terpinolene, cis-p-2-Menthen-1-ol, 1-Terpineol, Verbenone, α-Terpineol, trans-
Dihydrocarvone, trans-Piperitol, Nerol, cis-Sabinene hydrate acetate, trans-Sabinene
hydrate acetate, Isobornyl acetate, Nerylacetate, Geranylacetate, Caryophyllene,
α-Humulene
Leaves Essential oil Terpinoids cis-Sabinene hydrate, Terpinen-4-ol, γ-Terpinene, Linalyl acetate, trans-Sabinene Calín-Sánchez et al.
(dried) hydrate, Sabinene, α-Thujene, α-Pinene, Camphene, β-Myrcene, β-Pinene, 2-Hexen-1- (2015)
ol acetate, 3-Carene, α-Terpinene, p-Cymene, Limonene, cis-β-Ocimene, Terpinolene,
Linalool, cis-p-2-Menthen-1-ol, 1-Terpineol, Verbenone, α-Terpineol, trans-
Dihydrocarvone, trans-Piperitol, Nerol, cis-Sabinene hydrate acetate, trans-Sabinene
hydrate acetate, Isobornylacetate, Nerylacetate, Geranylacetate, Caryophyllene,
α-Humulene
Leaves Ethanolic extract Phenolic acids 3,4-dicaffeoylquinic acid, 4,5-dicaffeoylquinic acid, gallic acid, chlorogenic acid, Wahby et al. (2015)
tannic acid, cinnamic acid
Flavonoids Rutin, quercetin
Leaves Essential oil Terpinoids α-thujene, α-pinene, Sabinene, β-pinene, α-phellandrene, α-terpinene, p-cymene, El-Akhal et al.
β-phellandrene, γ-terpinene, Terpinolene, cis-Sabinene hydrate, trans-sabinene (2014)
hydrate, α-terpineol, Linalyle acetate, cis-sabinene hydrate acetate, Monoterpenes,
Hydrocarbon monoterpenes oxygenated, Monoterpenes, Trans dihydrocarvone, 4-
terpinene, p-menth-2-en-1-ol, β-caryophyllene, Valencene, α-humulene,
Monoterpenes, Monoterpenes hydrocarbon, Monoterpenes oxygenated,
Sesquiterpenes, Sesquiterpene hydrocarbon
Leaves Methanolic extract Phenolic acids Gallic acid, Chlorogenic acid, Caffeic acid, p-Coumaric acid, Ferulic acid, Rosmarenic Roby et al. (2013)
acid, Methyl rosmarenate
Flavonoids Apigenin, Luteolin-7-o-rutinose
Leaves Methanolic extract Total phenols nd Ayari et al. (2013)
Flavonoids nd
Tannins nd
Leaves Essential oil Terpinoids Terpinen-4-ol, γ-Terpinene, cis-sabinene-hydrate, α-Terpinene, α-Terpineol, trans- Jelali et al. (2011)
sabinene-hydrate, α-Terpinolene, Sabinene, Linalool, β-Pinene, p-Cymene, Linalyl
acetate
Leaves Essential oil Terpinoids Terpinene-4-ol, p-Cymene, α-Terpineol, Caryophyllene, Spathulenol, Caryophyllene Jiang et al. (2011)
oxide
Leaves Essential oil Terpinoids Trans-sabinene hydrate, Thymol, γ-terpinene, terpinen-4-ol, α-terpinene, limonene, Dambolena et al.
carvacrol, cis-sabinene hydrate, β-caryophyllene, α-terpineol, β-myrcene, carvacrol (2010)
methyl ether, germacrene D
Leaves Essential oil Terpinoids Terpinen-4-ol, γ-Terpinene, cis-sabinene-hydrate, α-terpinene, sabinene, terpinolene, Busatta et al. (2008)
α-thujene, α-pinene, β-pinene, myrcene, α-phellandrene, b-caryophyllene, p-cymene,
limonene, trans-p-menthenol, cis-p-menthenol, α-terpineol, trans-piperitol,
germacrene, spathulenol, caryophyllene oxide
Leaves Essential oil Terpinoids Tricyclene, α-Pinene, Camphene, Sabinene, α-Terpinene, p-Cimene, Limonene, 1,8- Banchio et al.
Cineole, γ-Terpinene, cis-Sabinene hydrate, Terpinen-4-ol, cis-Piperitol acetate, trans- (2008)
Piperitol acetate, Borneol, trans-Sabinene hydrate, α-Terpineol, trans-Carveol,
Camphor, β-Caryophyllene, α-Humulene
Leaves Essential oil Terpinoids Linalool, terpinen-4-ol, p-cymene, fenchone, sabinene, α-terpineol, geraniol, bornyl Charai et al. (1996)
acetate, lirnonene, β-caryophyllene, fenchol, α-campholenal
Leaves Essential oil Terpinoids cis-sabinene hydrate, terpinen-4-ol, γ-terpinene, α-terpineol, α-terpinene, trans- Arnold et al. (1993)
sabinene hydrate, linalool, sabinene, 1,8-cineole, lirnonene, terpinolene,
β-caryophyllene
Stems Essential oil Terpinoids α-Thujene, α-Pinene, Camphene, Sabinene, β-Pinene, 1-Octen-3-ol, 3-Octanone, Jan et al. (2018)
Myrcene, α-Phellandrene, 3-Carene, α-Terpinen, p-Cymene, Limonene,
β-Phellandrene, (Z)-β-Ocimene, γ-Terpinene, trans-Sabinene hydrate, Borneol,
Terpinen-4-ol, α-Terpineol, trans-Dihydrocarvone, Carvacrol methyl ether,
Thymoquinone, Thymol, Carvacrol, β-Caryophyllene, α-Humulene, Allo-
Aromadendrene, α-Muurolol, β-Bisabolene, γ-Cadinene, δ-Cadinene, Spathulenol,
Caryophyllene oxide, Epi-α-Muurolol, α-Eudesmol
Stems Essential oil Terpinoids Terpinen-4-ol, Thymol, trans-hydrate Sabineno, γ-Terpinene, r- Cymene, α-Terpineol, Guerra-Boone et al.
α-Terpinene, Thymol methyl ether, cis-hydrate Sabinene, Carvacrol methyl ether, (2015)
Terpinolene, (E)-β-Ocimene
Stems Methanolic extract Total phenols nd Ayari et al. (2013)
Flavonoids nt
Tannins nd
Stems Essential oil Terpinoids cis-sabinene hydrate, terpinen-4-ol, γ-terpinene, α-terpineol, α-terpinene, trans- Arnold et al. (1993)
sabinene hydrate, linalool, sabinene, 1,8-cineole, lirnonene, terpinolene,
β-caryophyllene
Flowers Essential oil Terpinoids α-Thujene, α-Pinene, Camphene, Sabinene, β-Pinene, 1-Octen-3-ol, 3-Octanone, Jan et al. (2018)
Myrcene, α-Phellandrene, 3-Carene, α-Terpinen, p-Cymene, Limonene,
β-Phellandrene, (Z)-β-Ocimene, γ-Terpinene, trans-Sabinene hydrate, Borneol,
Terpinen-4-ol, α-Terpineol, trans-Dihydrocarvone, Carvacrol methyl ether,
(continued on next page)

8
A. Bouyahya et al. Journal of Ethnopharmacology 265 (2021) 113318

Table 2 (continued )
Part used Extracts/essential oils Compounds Compounds References
groups

Thymoquinone, Thymol, Carvacrol, β-Caryophyllene, α-Humulene, Allo-

Aromadendrene, α-Muurolol, β-Bisabolene, γ-Cadinene, δ-Cadinene, Spathulenol,
Caryophyllene oxide, Epi-α-Muurolol, α-Eudesmol
Flowers Methanolic extract Total phenols nd Ayari et al. (2013)
Flavonoids nd
Tannins nd
Flowers Essential oil Terpinoids α-Thujene, α-Pinene, Sabinene, Myrcene, α-Terpinene, p-Cymene, β-Phellandrene, Novak et al. (2004)
γ-Terpinene, trans-Sabinene hydrate, α-Terpinolene, cis-Sabinene hydrate, Linalool,
cis-p-Menth-2-en-1-ol, trans-p-Menth-2-en-1-ol, Terpinen-4-ol, α-Terpineol, cis-
Sabinene hydrate acetate, Linaly acetate, β-Caryophyllene, α-Humulene,
Bicyclogermacrene
Flowers Essential oil Terpinoids Linalyl acetate, sabinene, γ-terpinene, cis-sabinene hydrate, cis-piperitol, α-terpineol, Barazandeh (2001)
p-cymene, (Z)-β-ocimene, β-caryophyllene, trans-sabinene hydrate, myrcene,
terpinolene
Flowers Essential oil Terpinoids cis-sabinene hydrate, terpinen-4-ol, γ-terpinene, α-terpineol, α-terpinene, trans- Arnold et al. (1993)
sabinene hydrate, linalool, sabinene, 1,8-cineole, lirnonene, terpinolene,
β-caryophyllene
Flowers Essential oil Terpinoids α-Terpinene, γ-Terpinene, Carvacrol, α-Pinene, β-Pinene Limonene, α-Thujene, Sarer et al. (1985)
α-Phellandrene, β-Phellandrene, Camphene, trans-β-ocimene, cis-β-ocimene, δ-4-
Carene, δ-3-Carene, p-Cymene, Myrcene, Terpinolene
Seeds Ethanol, methanol, acetone, Phenolic acids Catechol, Cinnamic acid, Gallic acid, Ascorbic acid Dhull et al. (2016)
and chloroform extracts
Seeds Essential oil Terpinoids cis-sabinene hydrate/linalool, Sabinene, trans-sabinene hydrate, α-terpineol, linalyl Novak et al. (2003)
acetate, limonene/β-phellandrene, bicyclogermacrene, β-caryophyllene, myrcene,
borneol, α-pinene
– Essential oil Terpinoids α-pinene, Sabinene, β-pinene, β-myrcene, α-phellandrene, α-terpinene, p-cymene, Xylia et al. (2019)
Sylvestrene, Eucalyptol, γ-terpinene, cis-Sabinene hydrate, Terpinolene, trans-
sabinene hydrate, cis-p-menth-2-en-1-ol, iso-3-Thujanol,Terpinen-4-ol, α-terpineol,
trans-Sabinene hydrate acetate, Linalool acetate, Bornyl acetate, Terpinene-4-ol
acetate, Caryophyllene E, Bicyclogermacrene
– Essential oil Terpinoids Terpinen-4-ol, γ-terpinene, α-terpinene Waller et al. (2019)
– Essential oil Terpinoids 1r-alpha-pinene, sabinene, cymol, cyclohexanol, cineole, alpha-terpinolene, alpha- Thanh et al. (2019)
terpineol, linalyl ester, beta-caryophyllene, alpha-caryophyllene
– Pure essential oil Terpinoids R-Pinene, Sabinene, Myrcene, 2-Carene, o-Cymene, Terpinyl acetate, Phellandrene, da Cunha et al.
Terpinene, Terpineol, (cis) Terpinolene, Linalool, Terpineol, trans-4-Isopropyl-1- (2018)
methyl-2-cyclohexen-1-ol, Terpinen-4-ol, ɑ-Terpineol, Linalyl anthranilate,
β-caryophyllene, Elixene
– Nanocapsules of essential oil Terpinoids β-citronellal, 3,7-dimethyl-2,6-octadien-1-ol, germacrene D, β-bourbonene, da Cunha et al.
β-caryophyllene,1,1,7-trimethyl-4-methylenedecahydro-1h-cyclopropa[e]-azulene, (2018)
torreyol, geranyl isobutyrate, 8,14-cedranoxide, spathulenol, caryophylene oxide,
cubenol, eudesmol, agaruspirol, β-eudesmol, widdrolhydroxyether, geranyltiglate, 1-
methylene-2b-hydroxymethyl-3,3-dimethyl-4b-(3-methylbut-2-enyl)-cyclohexane, 9-
(1-methylethylidene)-bicyclo[6.1.0]nonane, ethanol, 2-(3,3-
dimethylcyclohexylidene),
3-[2-(6-bromo-2-hydroxy-2,5,5,8 atetramethyldecahydro-1-naphthalenyl)ethyl]-3-
butene-1,2-diol,14-isopropyl-3,7,11-trimethyl-2,6,10-cyclotetradecatrien-1-
one,3,7,11-tetramethyltricyclo [5.4.0.0 (4,11)]undecan-1-ol,1,5,9,9-tetramethyl-2-
oxatricyclo [6.4.0.0 (4,8)] dodecane
– Essential oil Terpinoids α-thujene, α-pinene, camphene, sabinene, β-pinene, β-myrcene, α-phellandrene, Baj et al. (2018)
α-terpinene, p-cymene, limonene, 1.8-cineole, trans-β-ocimene, γ-terpinene, trans-
sabinene hydrate, α-terpinolene, linalool, cis-sabinene hydrate, fenchol, trans-p-2
menthen-1-ol, camphor, borneol, terpinen-4-ol, p-cymen-8-ol, α-terpineol, trans-
piperitol, linalyl acetate, α-terpineolacetate, eugenol, geranyl acetate, (E)-
β-caryophyllene, α-humulene, germacrene D, spathulenol, caryophyllene oxide
Flavonoids Luteolin-6,8-di-C-glucoside (lucenin-2), Apigenin-6,8-di-C-glucoside, (vicenin-2),
Luteolin-glucuronide, Apigenin-glucuronide, Apigenin
– Essential oil Terpinoids Benzenemethanol, 4-(1-methylethyl), Benzene, 1-methyl-2-(1-methylethyl), Sefeer and Elumalai
Cyclohexene, 1-methyl-4-(1-methylethylidene), 1,6-Octadien-3-ol, 3,7-dimethyl, (2018)
Caryophyllene, Bicyclo[3.1.0]hex-2-ene,2-methyl-5-(1-methylethyl), α–Pinene,
(+)-4-Carene, Camphene, Cyclohexene, 1-methyl-4-(5-methyl-1-methylene-4-hex­
enyl)-, (S), Benzene, 1-methoxy-4-methyl-2-(1-methylethyl)
– Essential oil Terpinoids Terpinen-4-ol, c-Terpinene, Linalool, p-Cymol, α-Pinene, Camphene, α-Terpineol, Muqaddas et al.,
α-Terpinene, β-Pinene, α-Terpinolene, β-Caryophyllene, Terpenyl acetate 2016
– Essential oil Terpinoids Terpinene-4-ol, γ-terpinene, Sabinene, cis-sabinene hydrate, α-Terpinene, α-terpineol, Abdalla and Hendi
trans-sabinene hydrate, p-cymene, Myrcene, trans-p-menthenol, cis-p-menthenol, (2014b)
α-Pinene, β-caryophyllene
– Essential oil Terpinoids 4-terpineol, γ-terpinene, trans-sabinene, α-terpinene, α-terpineol, Sabinene, p- Freire et al. (2011)
cymene, Limonene, cis-sabinene, trans-cariofilene
– Essential oil Terpinoids α-Pinene, Camphene, β-Myrcene, β-Pinene, α-Terpinene, p-Cymol, γ-Terpinene, Vagi et al. (2005)
Terpenyl acetate, α-Terpinolene, Linalool, Terpinen-4-ol, α-Terpineol,
β-Caryophyllene
– Ethanolic extract Terpinoids α-pinene, β-pinene, α-terpinene, p-cymol, γ-terpinene, α-terpinolene, linalool, Vagi et al. (2005)
terpenyl ester, terpinen-4-ol, α-terpineol, β-caryophyllene, neophytadiene,
spathulenol
– Supercritical fluid extract Terpinoids Vagi et al. (2005)
(continued on next page)

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A. Bouyahya et al. Journal of Ethnopharmacology 265 (2021) 113318

Table 2 (continued )
Part used Extracts/essential oils Compounds Compounds References
groups

α-terpinene, p-cymol, γ-terpinene, α-terpinolene, linalool, cis-sabinene hydrate,

terpenyl ester, terpinen-4-ol, α-terpineol, β-caryophyllene, neophytadiene,
spathulenol
– Essential oil Terpinoids cis-Sabinene hydrate, cis-Sabinene hydrate acetate, Sabinene, Linalyl acetate, Novak et al. (2002)
Linalool, trans-Sabinene hydrate, β-Caryophyllene, α-Terpineol, β-Phellandrene,
Bicyclogermacrene, Myrcene, Terpinen-4-ol, γ-Terpinene, β-Pinene
– Essential oil Terpinoids α-Thujene, α-Pinene, Camphene, Sabinene, β-Pinene, Myrcene, α-Phellandrene, Vera and
α-Terpinene, p-Cymene, Limonene, cis-Ocimene, trans-Ocimene, γ-Terpinene, Chane-Ming (1999)
Terpinolene, trans-sabinene hydrate, Linalool, cis-Sabinene hydrate, α-Pinene oxide,
trans-para-Menth-2en-1-ol, Camphor, cis-para-Menth-2-en-1-ol, Terpinen-4-ol,
α-Terpineol, trans-Piperitol, Isoborneol, cis-Piperitol, Nerol, Geraniol, Linalyl acetate,
Bornyl acetate, trans-Sabinyl acetate, Thymol, 5-Caranol, cis-dihydro-α-Terpinyl
acetate, α-Terpinyl acetate, Neryl acetate, Geranyl acetate, β-Caryophyllene,
α-Humulene, Bicyclogermacrene, Spathulenol, Caryophyllene oxyde, Acorenone

of O. majorana, from the qualitative and quantitative point of view, interesting results, in which Escherichia coli and Pseudomonas aeruginosa,
maybe due to certain ecological factors, to the part of the plant used, to which are known as highly resistant to antibacterial agents, were more
the age of the plant and the period of the vegetative cycle, or even to sensitive to the essential oil (MIC = 31.25 μg/mL) comparing to Staph­
genetic factors. ylococcus aureus.
Partovi et al. (2018) have evaluated the antibacterial effect of OMEO
(10% v/v) at a dose of 10 μL against foodborne bacteria using disc
3.6. Pharmacological investigation
diffusion, microdilution, and Time-Kill methods. And for the control,
chloramphenicol and streptomycin were used as positive control and
3.6.1. Antibacterial activity
dimethyl sulphoroxide (DMSO 2%) as negative control. As results,
Several works showed the antibacterial efficacy of different essential
OMEO exhibited an important antibacterial effect against Bacillus subtilis
oils and extracts from different plant parts. An investigation of previous
(33.16 ± 0.76 mm), Staphylococcus aureus (30.58 ± 0.72 mm), E. coli
researches shows that many studies listed antibacterial potential of
(23.58 ± 0.38 mm), Listeria monocytogenes (23.16 ± 0.38 mm), and
O. majorana against Gram-positive and Gram-negative bacteria (Charai
Salmonella enteric Typhimurium (22.96 ± 0.25 mm) more than the
et al., 1996; Leeja and Thoppil, 2007; Busatta et al., 2008; Choi and
positive controls namely streptomycin and chloramphenicol. The
Rhim, 2008; de Oliveira et al., 2009; Ramos et al., 2011; Omara et al.,
microdilution assay indicates that OMEO exhibits a bacteriostatic effect
2014; Adam and Ahmed, 2014; de Lima Marques et al., 2015; Lakhrissi
against Bacillus subtilis, Listeria monocytogenes, Salmonella enterica
et al., 2015; Ouedrhiri et al., 2016; Partovi et al., 2018; Amor et al.,
Typhimurium, and Staphylococcus aureus, while exhibits bactericidal
2019; Al-Fatimi, 2018; Chaves et al., 2019).
effect against Escherichia coli. According to Time-Kill assay, the OMEO
The antibacterial activity of OMEO as well as its extracts from
possesses a bactericidal effect against B. subtilis (at 3 h), Staphylococcus
different parts (aerial parts and leaves) using inhibition zone diameter
aureus, and Listeria monocytogenes (at 6 h). While OMEO showed a
(IZD) and/or minimum inhibitory concentration (MIC) methods are
bactericidal effect against Salmonella enterica Typhimurium and
summarized in Table 3.
Escherichia coli at 24 h. In another work, the agar diffusion method
Charai et al. (1996) have tested in vitro the antibacterial effect of
demonstrated that OMEO from leaves inhibits the growth of Staphylo­
Origanum majorana essential oil (0.1, 0.2, 0.4, 0.8, 1.6, 2, 4, and 5 ppm)
coccus aureus (16 mm), Escherichia coli (15 mm), Klebsiella pneumoniae
and its aqueous extract (1, 4, 25, and 40%) obtained from the plant
(13 mm), and Enterococcus faecalis (12 mm) compared to positive con­
leaves. This study showed that OMEO exhibits a total inhibition at 5 ppm
trols; Ampicillin (10 μg), Amikacine (30 μg), Piperaciline (8 μg), and
against Pseudomonas fluorescens T32, Pseudomonas fluorescens T3,
negative control dimethyl sulphoroxide (DMSO 2%) (Ramos et al.,
Escherichia coli 1 (EC1), Escherichia coli 7 (EC7), Staphylococcus aureus 4
2011).
(S4). In another study, de Lima Marques et al. (2015) have evaluated the
More interestingly, the antibacterial activity of O. majorana EO
antibacterial activity of OMEO (10 μL) extracted from the leaves against
(1.2%) at a dose of 5 μL was determined in vitro and in a food system
Staphylococcus aureus using disk diffusion and microdilution methods,
against foodborne bacteria (Busatta et al., 2008). The results showed
and compared with Tween 80 as negative control and six different an­
that among 19 microorganisms, tested by the disc diffusion method and
tibiotics (Ampicilin (10 μg), Ciprofloxacin (5 μg), Chloramphenicol (30
using chloramphenicol (30 mg) as positive control, 10 strains, namely,
μg), Gentamicin (10 μg), Sulphazotrin (25 μg), and Tetracyclin (10 μg) as
Bacillus subtilis, Enterococcus faecalis, Staphylococcus aureus, and Strep­
positive control. The results showed that OMEO exhibits an
tococcus mutans (Gram Positive), and Aeromonas sp., Escherichia coli,
anti-Staphylococcal activity with an inhibitory zone of 41 mm and MIC
Klebsiella pneumonia, Salmonella choleraensius, Serratia sp., and Shigella
equal to 50 μl/mL− 1. In another study, Lakhrissi et al. (2015) have
flexneri (Gram negative) were sensitive and selected to determine the
studied synergetic effect of OMEO and Origanum vulgare essential oils
minimal inhibitory concentration. The microdilution approach indicates
(OVEO) against Staphylococcus aureus (Gram positive) and Klebsiella
that all the microorganisms selected and tested were sensitive to the
pneumoniae, Escherichia coli, Escherichia cloacae, Pseudomonas aeruginosa,
action of OMEO with MIC values varied from 0.069 to 2.3 mg/mL. The
and Acinetobacter sp. (Gram negative). This study demonstrated that,
antibacterial effect of the highest MIC in a food system comprising fresh
except Pseudomonas aeruginosa, all bacteria were sensitive to the mixture
sausage during storage showed that their addition causes bactericidal
obtained from OMEO/OVEO, while the most sensitive bacteria was
action at a concentration of 5.75 mg/g and bacteriostatic at a concen­
Acinetobacter sp. (25 mm) followed by Staphylococcus aureus (22 mm),
tration of 1.15 mg/g against Escherichia coli (Busatta et al., 2008).
Escherichia coli and Klebsiella pneumoniae (20 mm), and Escherichia
In addition, the antibacterial effect of OMEO from the aerial part was
cloacae (18 mm). Chaves et al. (2019) have tested the antibacterial effect
evaluated by many studies. Indeed, Amor et al. (2019) have evaluated in
of O. majora EO at a concentration of 2000 μg/mL against Staphylococcus
vitro the antibacterial activity of the essential oil of O. majorana aerial
aureus, Pseudomonas aeruginosa, and Escherichia coli using standard an­
parts (0.97%) at a dose of 5–50 μL against Gram positive and Gram
tibiotics amoxicillin (50 μg/mL) as positive control and dimethyl sul­
negative bacteria and used ethanol as negative control and Tetracycline
phoroxide (DMSO 2%) as negative control. The author reported

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A. Bouyahya et al. Journal of Ethnopharmacology 265 (2021) 113318

Fig. 2. Structure of phenolic acids compounds identified in O. majorana.

11
A. Bouyahya et al. Journal of Ethnopharmacology 265 (2021) 113318

Fig. 3. Structure of flavonoids compounds identified in O. majorana.

(10 μg) and Gentamycin (10 μg) as positive controls. The results showed varied between 6.0 ± 0.0 and 32.3 ± 2.5 mm. Among the 20 microor­
that OMEO inhibits the bacterial growth in a concentration-dependent ganisms tested, the diameter of the inhibition zones of 10 bacteria were
manner on all microorganisms tested. The important antibacterial ef­ superior to 20 mm, while the most sensitive bacteria was Staphylococcus
fect was determined at 50 μL, while the diameter of the inhibition zones aureus (32.3 ± 2.5 mm), followed by Salmonella enterica Typhimurium

12
A. Bouyahya et al. Journal of Ethnopharmacology 265 (2021) 113318

Fig. 4. Structure of terpenoids compounds identified in O. majorana.

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A. Bouyahya et al. Journal of Ethnopharmacology 265 (2021) 113318

Fig. 4. (continued).

(29.7 ± 0.6 mm), Bacillus clausii (28.3 ± 1.5 mm), Escherichia coli (26.7 bactericidal effect against Klebsiella spp. at 4h, Proteus spp. at 8h, and
± 0.6 mm), Brochothrix thermosphacta (26.3 ± 1.2 mm), Streptococcus S. aureus at 24h. Another study reported the antibacterial activity of
salivarius (24.3 ± 1.2 mm), Staphylococcus saprophyticus (24.3 ± 1.2 O. majorana EO (2.3%) at a dose of 10 μL extracted from its aerial part
mm), and Hafnia alvei (20.3 ± 0.6 mm). The evaluation of the antibac­ against four Gram-positive bacteria (Staphylococcus aureus, Staphylo­
terial activity of the essential oil of O. majorana aerial parts against 12 coccus epidermidis, Streptococcus aureus, and Enterococcus faecalis) and
strains using the disc diffusion and microdilution techniques showed six Gram-negative bacteria (Escherichia coli, Pseudomonas aeruginosa,
that the oil was active against all bacterial strains tested. In fact, the Klebsiella pneumoniae, Proteus mirabilis, Shigella dysenteria, and Salmo­
diameter of the inhibition zones varied from 8 ± 0 to 18.33 ± 0.57 mm. nella enteritidis) (Omara et al., 2014). Indeed, the results report that the
On the other hand, the MBC/MIC ratio indicates that O. majorana EO most sensitive bacteria were Escherichia coli, Streptococcus aureus,
exhibits bactericidal action against Salmonella enterica Typhimurium, Shigella dysenteria, and Salmonella enteritidis with the same MIC value
Vibrio parahaemolyticus, Pseudomonas aeruginosa, Staphylococcus epi­ (1/1256). The less sensitive one was Pseudomonas aeruginosa (MIC =
dermidis, Staphylococcus aureus, and Micrococcus luteus. While a bacte­ 1/16).
riostatic effect was exhibited on the rest of the bacteria tested. Ouedrhiri Earlier works listed the O. majorana extract antibacterial efficacy.
et al. (2016) have tested the antibacterial activity of OMEO (1.2%) at a Adam and Ahmed (2014) have investigated the antibacterial activity of
dose of 10 μL against Staphylococcus aureus, Bacillus subtilis, Escherichia petroleum ether and mixture methanol-water extracts (50%, 25%, and
coli, Pseudomonas aeruginosa. In this study, the most sensitive strain was 12.5%) obtained from the aerial part of O. majorana using the disc
Escherichia coli (20.33 ± 2.30 mm), followed by Bacillus subtilis (19.66 ± diffusion method and dimethyl sulphoroxide (0.1 mL) as negative con­
1.52 mm), Staphylococcus aureus (16.33 ± 2.51 mm), and Pseudomonas trol and Gentamicine (0.1 mL) as positive control. This study concluded
aeruginosa (9.66 ± 0.57 mm). Also, a bactericidal effect was exhibited by that Escherichia coli was the most sensitive bacteria to the petroleum
O. majorana EO against Staphylococcus aureus, Escherichia coli, and Ba­ ether extract (21 mm), followed by Pseudomonas aeruginosa and Staph­
cillus subtilis. In addition, this study demonstrated that the synergy ylococcus aureus with 16 mm and 15 mm, respectively. The same strain
optimal mixture against Escherichia coli was composed by O. majorana was the most sensitive to methanol-water extract (15 mm), followed by
and O. compactum essential oils at 25% and 75%, respectively. The study Pseudomonas aeruginosa and Staphylococcus aureus with 15 mm and 14
reported by de Oliveira et al. (2009) indicates that O. majorana EO (160, mm, respectively. In another study, Leeja and Thoppil (2007) reported a
80, 40, 20, 10, 5, and 2.5 μL/mL) at dose of 20 μL possesses an inter­ good antibacterial activity of O. majorana methanol extract against Ba­
esting antibacterial activity against six bacteria isolated from patients cillus subtilis, Bacillus megaterium, Escherichia coli, Proteus vulgaris, Pseu­
with conjunctivitis. In fact, the diameter of the inhibitory zones varied domonas aeruginosa, and Staphylococcus aureus compared to Gentamycin
between 15 and 20 mm with MIC values ranged from 2.5 to 10 μL/mL. In (40 mg/mL) used as positive control. The methanol extract inhibits the
addition, the “Time Kill study” indicates that O. majorana EO exhibits a growth of all bacteria with diameters of the inhibition zones ranged from

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A. Bouyahya et al. Journal of Ethnopharmacology 265 (2021) 113318

Table 3
Antibacterial effects of O. majorana essential oils and extracts.
Part used Extracts Tested strains Key results References

Aerial parts Essential oil (0.97%) Gram Positive Ф = 28.3 ± 1.5 mm Amor et al. (2019)
Dose (50 μL) Bacillus Clausii 2226
Brochothrix Thermosphacta 274 Ф = 26.3 ± 1.2 mm
Brochothrix Thermosphacta 7R1 Ф = 21.3 ± 0.6 mm
Carnobacterium maltaromaticum Ф = 9.3 ± 0.6 mm
9P
Carnobacterium maltaromaticum Ф = 9.7 ± 0.6 mm
H1201
Carnobacterium maltaromaticum Ф = 13.3 ± 1.5 mm
D1203
Carnobacterium maltaromaticum Ф = 6.0 ± 0.0 mm
F1201
Escherichia coli 32 Ф = 26.7 ± 0.6 mm
Enterococcus faecalis 226 Ф = 9.7 ± 0.7 mm
Enterococcus faecalis E21 Ф = 6.0 ± 0.0 mm
Listeria innocua 1770 Ф = 13.7 ± 1.5 mm
Staphylococcus aureus Ф = 32.3 ± 2.5 mm
Streptococcus salivarius Ф = 24.3 ± 1.2 mm
Staphylococcus 3S Ф = 24.3 ± 1.2 mm
Staphylococcus sp. ES1 Ф = 21.0 ± 1.0 mm
Staphylococcus sp. GB1 Ф = 19.3 ± 1.2 mm
Gram negative Ф = 13.7 ± 1.5 mm
Hafnia alvei 53M
Pseudomonas fragi 6P2 Ф = 9.3 ± 0.6 mm
Salmonella enterica Ф = 29.7 ± 0.6 mm
Typhimurium
Streptococcus proteamaculans Ф = 19.3 ± 1.2 mm
20P
Leaves Essential oil Gram Positive MIC = 0.069 mg/mL Busatta et al. (2008)
Dose (5 μL) Bacillus subtilis
Enterococcus faecalis MIC = 2.300 mg/mL
Staphylococcus aureus MIC = 0.782 mg/mL
Streptococcus mutans MIC = 2.300 mg/ML
Gram negative MIC = 0.920 mg/mL
Aeromonas sp.
Escherichia coli MIC = 0.920 mg/mL
Klebsiella pneumoniae MIC = 0.920 mg/mL
Salmonella choleraensius MIC = 0.920 mg/mL
Serratia sp. MIC = 2.300 mg/mL
Shigella flexneri MIC = 0.782 mg/mL
Aerial parts Essential oil (12, 6, 3 mg/mL) Gram-positive Inhibit the growth of bacteria at concentration of Della Pepa et al. (2019)
Dose (10 μL) Bacillus megaterium 12 mg/mL
Clavibacter michiganensis Inhibit the growth at all tested concentrations
Gram-negative Inhibit the growth of bacteria at concentration of
Xanthomonas campestris 12 mg/mL
Pseudomonas fluorescens Inhibit the growth of bacteria at concentration of
12 mg/mL
Pseudomonas syringae pv. Inhibit the growth of bacteria at concentration of 6
Phaseolicola mg/mL
Aerial parts Essential oil (50 mg/mL) Gram positive MIC = 0.39 mg/mL, MBC = 1.562 mg/mL Hajlaoui et al., 2016
Dose (10 μL) Staphylococcus epidermidis CIP
106510
Staphylococcus aureus CIP MIC = 0.195 mg/mL, MBC = 0.781 mg/mL
106510
Micrococcus luteus NCIMB 8166 MIC = 0.097 mg/mL, MBC = 0.39 mg/mL
Enterococcus feacalis 29212 MIC = 0.39 mg/mL, MBC = 3.125 mg/mL
Bacillus cereus ATCC 11778 MIC = 0.195 mg/mL, MBC = 1.562 mg/mL
Bacillus cereus ATCC 14579 MIC = 0.097 mg/mL, MBC = 0.781 mg/mL
Gram negative MIC = 0.781 mg/mL, MBC = 6.25 mg/mL
Escherichia coli ATCC 35218
Listeria monocytogenes MIC = 0.39 mg/mL, MBC = 3.125 mg/mL
Pseudomonas aeruginosa MIC = 3.125 mg/mL, MBC = 12.5 mg/mL
Salmonella typhimurium LT2 MIC = 1.562 mg/mL, MBC = 3.125 mg/mL
DT104
Vibrio parahaemolyticus MIC = 1.562 mg/mL, MBC = 6.25 mg/mL
Vibrio alginolyticus ATCC 17749 MIC = 0.39 mg/mL, MBC = 3.125 mg/mL
Ethanolic extract (0.2, 0.4% (m/v)) Gram positive Inhibitory effect = 76.9 ± 7.9% at concentration of Vagi et al. (2005)
Bacillus cereus 0.4% (m/v)
Gram negative Inhibitory effect = 21.5 ± 0.3% at concentration of
Escherichia coli 0.4% (m/v)
Pseudomonas fluorescens Inhibitory effect = 56.6 ± 16.4% at concentration
of 0.4% (m/v)
Supercriticalfluid extract (0.2, 0.4% (m/ Gram positive Inhibitory effect = 98.1 ± 1.7% at concentration of Vagi et al. (2005)
v)) Bacillus cereus 0.4% (m/v)
(continued on next page)

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A. Bouyahya et al. Journal of Ethnopharmacology 265 (2021) 113318

Table 3 (continued )
Part used Extracts Tested strains Key results References

Gram negative Inhibitory effect = 96.4 ± 6.3% at concentration of

Escherichia coli 0.4% (m/v)
Pseudomonas fluorescens Inhibitory effect = 98.8 ± 2.1% at concentration of
0.4% (m/v)
Aerial parts Essential oil Gram positive Ф = 16.33 ± 2.51 mm, Ouedrhiri et al. (2016)
Dose (10 μL) Staphylococcus aureus MIC = 0.25%, MBC = 2% (v/v)
Bacillus subtilis Ф = 19.66 ± 1.52 mm,
MIC = 0.125%, MBC = 2% (v/v)
Gram negative Ф = 20.33 ± 2.30 mm,
Escherichia coli MIC = 0.25%, MBC = 0.25% (v/v)
Pseudomonas aeruginosa Ф = 9.66 ± 0.57 mm,
MIC = 2%, MBC= >4% (v/v)
Essential oil Gram positive Ф = 21.20 ± 1.24 mm Almasi et al. (2020)
Dose (100 μL) Staphylococcus aureus
Gram negative Ф = 15.10 ± 1.09 mm
Escherichia coli
Essential oil Gram positive Ф = 41 mm, MIC = 50 μg/mL, MBC > CMT de Lima Marques et al.
Dose (10 μL) Staphylococcus aureus (2015)
Areal parts Petroleum ther (50%, 25%, and 12.5%) Gram positive Ф = 15 mm Adam and Ahmed
Staphylococcus auries (2014)
Gram negative Ф = 21 mm
Escherichia coli
Pseudomonas aeruginosa Ф = 16 mm
Areal parts Methanol water (50%, 25%, and 12.5%) Gram positive Ф = 14 mm Adam and Ahmed
Staphylococcus auries (2014).
Gram negative Ф = 19 mm
Escherichia coli
Pseudomonas aeruginosa Ф = 15 mm
Aerial parts Essential oil (250 μg/mL) Gram positive MIC = From 150.0 to 250.0 μg/mL Selim et al. (2013)
Staphylococcus aureus
Bacillus cereus MIC = From 75.0 to 150.0 μg/mL
Gram negative MIC = From 150.0 to 250.0 μg/mL
Salmonella indica
Escherichia coli MIC = Over 250.0 μg/mL
Leaves Essential oil Gram positive Ф = 16 mm Ramos et al. (2011)
Dose (10 μL) Staphylococcus aureus (ATCC
25923)
Enterococcus faecalis (ATCC, Ф = 12 mm
19433)
Gram negative Ф = 15 mm
Escherichia coli (ATCC 25992)
Klebsiella pneumoniae (ATCC Ф = 13 mm
23357)
Leaves Essential oil Gram positive Ф = 33.16 ± 0.76 mm Partovi et al. (2018)
Dose (10 μL) Listeria monocytogenes ATCC
7644
Staphylococcus aureus ATCC Ф = 30.58 ± 0.72 mm
65138
Bacillus subtilis ATCC 11778 Ф = 33.16 ± 0.76 mm
Gram negative Ф = 23.58 ± 0.38 mm
Escherichia coli O157:H7
Salmonella typhimurium ATCC Ф = 22.96 ± 0.25 mm
14028
Aerial parts Essential oil Gram positive Ф = 16.75 mm Omara et al. (2014)
Shigella
Salmonella Ф = 19.20 mm
Pseudomonas Ф = 14.00 mm
Proteus Ф = 15.00 mm
Escherichia coli Ф = 19.00 mm
Citrobacter Ф = 16.00 mm
Salmonella typhimurium ATCC Ф = 13.00 mm
14028
Essential oil (160, 80, 40, 20, 10, 5, and Gram positive Ф = 22 mm de Oliveira et al. (2009)
2.5 μL/mL) Staphylococcus aureus BH01 MIC = 5 μL/mL
Dose (20 μL) Staphylococcus aureus BH02 Ф = 20 mm
MIC = 10 μL/mL
Staphylococcus aureus BH3 Ф = 21 mm
MIC = 2.5 μL/mL
Staphylococcus coagulase- BH8 Ф = 18 mm
MIC = 5 μL/mL
Staphylococcus coagulase + BH9 Ф = 20 mm
MIC = 10 μL/mL
Gram negative Ф = 15 mm
Enterobacter spp. BH15 MIC = 2.5 μL/mL
Areal parts Essential oil (0.25–50 μg/mL) Gram positive Ф = 8 ± 0.04 mm Olfa et al. (2016)
Dose (5 μL) Enterococcus faecalis
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A. Bouyahya et al. Journal of Ethnopharmacology 265 (2021) 113318

Table 3 (continued )
Part used Extracts Tested strains Key results References

Paenibacillus larvae Ф = 7 ± 0.01 mm

Staphylococcus aureus Ф = 9 ± 0.01 mm
Listeria monocytogenes Ф = 11 ± 0.02 mm
Listeria ivanovii Ф = 12 ± 0.11 mm
Listeria inocua Ф = 10 ± 0.12 mm
Gram negative Ф = 16 ± 0.03 mm
Escherichia coli
Salmonella enteritidis Ф = 12 ± 0.05 mm
Aerial parts Methanol extract Gram positive Ф = 23 mm Leeja and Thoppil
Staphylococcus aureus (2007)
Bacillus subtilis Ф = 42 mm
Bacillus megaterium Ф = 20 mm
Gram negative Ф = 20 mm
Proteus vulgaris
Pseudomonas aeruginosa Ф = 18 mm
Escherichia coli Ф = 18 mm
Leaves Essential oil Gram positive Ф = 22 mm Lakhrissi et al. (2015)
Staphylococcus
Gram negative Ф = 20 mm
Klebsiella pneumoniae
Acinetobacter Ф = 25 mm
Pseudomonas No inhibition
Escherichia coli Ф = 20 mm
Enterobacter Ф = 18 mm
Ethanolic extract Gram positive Ф = 12 mm Joshi et al. (2009)
Dose (50 μL) Staphylococcus aureus MBC = 2.5 mg/mL
Bacillus subtilis Ф = 14 mm
MBC = 2.5 mg/mL
Bacillus cereus Ф = 10 mm
MBC>10 mg/mL
Bacillus thuringiensis Ф = 8 mm
MBC = 5 mg/mL
Gram negative No effect
Klebsiella pneumoniae
Proteus spp No effect
Escherichia coli No effect
Pseudomonas spp No effect
Salmonella Typhi No effect
Shigella dysenteriae No effect
Essential oil (0.1, 0.5, 1, 5, 10, 20, 30, 40, Gram positive Inhibition halo = 1.6 cm at concentration of 50% Freire et al. (2011)
and 50%) Staphylococcus aureus
Gram negative Inhibition halo = 1.6 cm at concentration of 50%
Escherichia coli
Aerial parts Dichloromethane extract Gram positive Ф = 20 mm Al-Fatimi (2018)
Bacillus subtilis
Staphylococcus aureus Ф = 30 mm
Micrococcus flavus Ф = 15 mm
Gram negative Ф = 15 mm
Pseudomonas aeruginosa
Escherichia coli Ф = 20 mm
Aerial parts Methanolic extract Gram positive Ф = 15 mm Al-Fatimi (2018)
Bacillus subtilis
Staphylococcus aureus Ф = 35 mm
Micrococcus flavus Ф = 20 mm
Gram negative Ф = 15 mm
Escherichia coli
Pseudomonas aeruginosa Ф = 20 mm
Aerial parts Aqueous extract Gram positive Ф = 10 mm Al-Fatimi (2018)
Bacillus subtilis
Staphylococcus aureus Ф = 15 mm
Micrococcus flavus Ф = 10 mm
Gram negative Ф = 10 mm
Pseudomonas aeruginosa
Escherichia coli Ф = 10 mm
Aerial parts Essential oil Gram positive Ezzeddine et al. (2001)
Staphylococcus aureus MBC = 1/128
Staphylococcus epidermitis MBC = 1/128
Enterococcus faecalis MBC = 1/128
Streptococcus A MBC = 1/256
Gram negative MBC = 1/16
Pseudomonas aeruginosae
Klebsiella pneumoniae MBC = 1/128
Proteus mirabilis MBC = 1/128
Shigella dysenteria MBC = 1/256
Escherichia coli MBC = 1/256
Salmonella enteritidis MBC = 1/256
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A. Bouyahya et al. Journal of Ethnopharmacology 265 (2021) 113318

Table 3 (continued )
Part used Extracts Tested strains Key results References

Leaves and Essential oil (1.0–10 μL/mL) Gram positive Ф = 5.5 mm Deans and Svoboda
stems Staphylococcus aureus (1990)
Micrococcus iuteus Ф = 10.6 mm
Leuconostoc cremoris Ф = 8.1 mm
Lactobacillus plantarum Ф = 17.7 mm
Enterococcus faecalis Ф = 8.7 mm
Enterobacter aerogenes Ф = 11.6 mm
Clostridiuni sporogenes Ф = 8.7 mm
Brocothrix thermosphacta Ф = 8.3 mm
Brevibacterium linens Ф = 13.9 mm
Bacillus subtilis Ф = 13.6 mm
Gram negative Ф = 20.1 mm
Beneckea natriegens
Alcaligenes faecalis Ф = 11.1 mm
Aeromonas hydrophila Ф = 11.5 mm
Citrobacter freundii Ф = 10.0 mm
Erwinia carotovora Ф = 11.6 mm
Escherichia coli Ф = 8.7 mm
Flavobacterium suaveolens Ф = 22 .4 mm
Klebsiella pneumoniae Ф = 9.4 mm
Moraxella sp. Ф = 21.7 mm
Proteus vulgaris Ф = 12.9 mm
Pseudomonas aeruginosa Ф = 19.2 mm
Salmonella pullorum Ф = 14.3 mm
Serratia marcescens Ф = 12.1 mm
Acinetobacter calcoacetica Ф = 14.8 mm
Yersinia enterocolitica Ф = 15.7 mm
Leaves Essential oil (2000 μg/mL) Gram positive MIC = Not present Chaves et al. (2019)
Staphylococcus auereus MBC = Not present
Gram negative MIC = 31.25 μg/mL
Pseudomonas aeruginosa MBC = 1000 μg/mL
Escherichia coli MIC = 31.25 μg/mL
MBC = 500 μg/mL
Ethanol extract Gram positive Ф = 31.8 ± 0.29 mm Choi and Rhim (2008)
Staphylococcus aureus KCTC
1928
Listeria monocytogenes ATCC Ф = 30.5 ± 0.45 mm
19117
Gram negative Ф = 32.0 ± 0.50 mm
Salmonella enteritidis
Salmonella cholerasuis ATCC Ф = 30.1 ± 0.36 mm
2931
Salmonella paratyphi Ф = 31.8 ± 0.25 mm
Escherichia coli KCTC 2441 Ф = 29.5 ± 0.35 mm
Escherichia coli 0157:H7 ATCC Ф = 31.1 ± 0.32 mm
43890
Yersinia enterocolitica ATCC Ф = 30.6 ± 0.32 mm
23715
Salmonella typhi KCCM 11808 Ф = 31.4 ± 0.53 mm
Salmonella typhimurium KCTC Ф = 31.8 ± 0.72 mm
2491
Leaves Aqueous extract (1, 4, 25, and 40%) Gram positive No inhibition Charai et al. (1996)
Staphylococcus aureus (S4) No inhibition
Bacillus cereus 24 No inhibition
Bacillus cereus 30 No inhibition
Gram negative No inhibition
Pseudomonas fluorescens T2 No inhibition
Pseudomonas fluorescens T3 No inhibition
Escherichia coli EC1
Escherichia coli EC7
Leaves Essential oil (0.1, 0.2, 0.4, 0.8, 1.6, 2, 4, Gram positive Inhibition at concentration of 1 ppm Charai et al. (1996)
and 5 ppm) Staphylococcus aureus (S4) Inhibition at concentration of 0.4 ppm
Bacillus cereus 24 Inhibition at concentration of 0.4 ppm
Bacillus cereus 30 No inhibition
Gram negative Inhibition at concentration of 0.8 ppm
Pseudomonas fluorescens T2 Inhibition at concentration of 2 ppm
Pseudomonas fluorescens T3 Inhibition at concentration of 1.6 ppm
Escherichia coli EC1
Escherichia coli EC7

18 to 42 mm. The most sensitive bacteria was Bacillus subtilis (42 mm), aerial parts tested by disc diffusion method and used ampicillin (10
followed by Staphylococcus aureus (23 mm), Bacillus megaterium, Proteus μg/disc) and gentamicin (10 mg/disc) as positive controls. According to
vulgaris (20 mm), Escherichia coli, and Pseudomonas aeruginosa with same this work, the dichloromethane and methanol extracts exhibited more
the diameter of 18 mm. Al-Fatimi (2018) studied the antibacterial ac­ qualitative and quantitative activity than aqueous extracts against
tivity of dichloromethane, methanol, and water extracts of O. majorana bacterial strains tested. In fact, the disc diffusion method demonstrated

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Table 4 Table 4 (continued )

Antifungal activity of O. majorana essential oils and extracts. Part Extracts Tested strains Key results References
Part Extracts Tested strains Key results References used
used
MFC = 0.937
Aerial Essential oil Aspergillus niger Ф = 14.0 ± 1.0 Amor et al. mg/mL
parts (0.97%) mm (2019) Microsporum Ф = 12 ± 0 mm
Dose (20 μL) canis MIC = 0.234
Aerial Essential oil Penicillium Important Della Pepa mg/mL
parts (2000, 1000, expansum antifungal et al. MFC = 1.875
and 500 ppm) Aspergillus niger activity at (2019) mg/mL
Monilinia concentration of Essential oil (80 Sporothrix MIC≤ 0.07–2.25 Waller
fructicola 2000 ppm mg/kg) brasiliensis mg/mL et al.
Aerial Essential oil (30 Candida Ф = 28 ± 0 mm Hajlaoui (2019)
parts mg/mL) albicans ATCC MIC = 0.058 et al., 2016 Essential oil Sporothrix No inhibition Waller
90028 mg/mL (72–2.25 mg/ brasiliensis No inhibition et al.
MFC = 0.234 mL) (Humans) (2016)
mg/mL Dose (100 μL) Sporothrix MIC90 ≤ 2.25
Candida Ф = 15.33 ± brasiliensis mg/mL
glabrata ATCC 0.58 mm (Dogs) MFC90 = 9 mg/
90030 MIC = 0.117 mL
mg/mL Sporothrix MIC90 = 4.5 mg/
MFC = 0.468 brasiliensis mL
mg/mL (Cats) MFC90 = 9 mg/
Candida Ф = 14.33 ± mL
parapsilosis 0.58 mm Sporothrix MIC90 = 4.5 mg/
ATCC 22019 MIC = 0.117 brasiliensis mL
mg/mL (Overall) MFC90 = 9 mg/
MFC = 0.234 mL
mg/mL Sporothrix No inhibition
Candida krusei Ф = 13 ± 0.0 mm schenckii (IOC No inhibition
ATCC 6258 MIC = 0.117 1226)
mg/mL Sporothrix MIC90 = 4.5 mg/
MFC = 0.468 schenckii mL
mg/mL (Overall) MFC90 = 9 mg/
Saccharomyces Ф = 11 ± 0.0 mm mL
cerevisae MIC = 0.468 Ethanolic Aspergillus niger Inhibitory effect Vagi et al.
mg/mL extract (2.5% = 19.3 ± 12.5% (2005)
MFC = 0.937 w/v) at the
mg/mL concentration of
Candida Ф = 13.66 ± 2.5% (w/v)
tropicalis 0.57 mm Trichoderma Inhibitory effect
MIC = 0.234 viride = 0% at the
mg/mL concentration of
MFC = 0.937 2.5% (w/v)
mg/mL Penicillium Inhibitory effect
Candida Ф = 12.33 ± cyclopium = 39.8 ± 4.3% at
glabrata 0.57 mm the
MIC = 0.468 concentration of
mg/mL 2.5% (w/v)
MFC = 0.937 Supercritical Aspergillus niger Inhibitory effect Vagi et al.
mg/mL fluid extract = 100% at the (2005)
Candida Ф = 11.33 ± (0.05, 0.15, concentration of
albicans 0.57 mm 0.25, 0.4, and 0.4% (w/v)
MIC = 0.468 0.5% w/v) Trichoderma Inhibitory effect
mg/mL viride = 100% at the
MFC = 1.875 concentration of
mg/mL 0.5% (w/v)
Candida Ф = 12 ± 0 mm Penicillium Inhibitory effect
Parapsilosis MIC = 0.234 cyclopium = 100% at the
mg/mL concentration of
MFC = 0.937 0.4% (w/v)
mg/mL Essential Oil Phytophthora Ф = 18.00 ± Thanh
Trichophyton Ф = 11.66 ± (500, 1000, infestants 0.50 mm at the et al.
violaceum 0.57 mm 2000, and 5000 concentration of (2019)
MIC = 0.468 ppm) 5000 ppm
mg/mL Aerial Essential oil (22 Aspergillus niger Fungal growth Salha et al.
MFC = 1.875 parts mg/mL) inhibition = (2017)
mg/mL Dose (5 μL) 36.56 ± 9.56% at
Trichophyton Ф = 11 ± 0 mm the
rubrum MIC = 0.234 concentration of
mg/mL 22 mg/mL
MFC = 1.875 Aerial Petroleum ether Candida No inhibition Adam and
mg/mL parts (12.5%, 25%, albicans zone Ahmed
Trichophyton Ф = 15.33 ± and 50%) Aspergillus niger (2014)
mentagrophytes 0.57 mm Methanol + Candida No inhibition
MIC = 0.117 water extract albicans zone
mg/mL (12.5%, 25%, Aspergillus niger
and 50%)
(continued on next page)

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A. Bouyahya et al. Journal of Ethnopharmacology 265 (2021) 113318

Table 4 (continued ) Vagi et al., 2005; Adam and Ahmed, 2014; Hajlaoui et al., 2016; Waller
Part Extracts Tested strains Key results References et al., 2019). The previous results of the antifungal activity of
used O. majorana EOs and its extracts are summarized in Table 4.
Aerial Essential oil Verticillium Ф = 14 mm Rus et al.
The investigation of the antifungal activity of OMEO has started
parts (0.25 mg/L, 0.5 dahliae MIC = 5 mg/L (2015) since 1990 by Deans and Svoboda (1990). This study reports the anti­
mg/L, 1 mg/L, MFC = 20 mg/L fungal effect of OMEO, at concentrations ranging from 1.0 to 10 μL/mL,
5 mg/L, 10 mg/ Penicillium Ф = 10 mm against Aspergillus flavus, Aspergillus niger, Aspergillus ochraceus, Asper­
L, 15 mg/L, 20 aurantiogriseum MIC = 1 mg/L
gillus parasiticus, and Trichoderma viride. The results showed that OMEO
mg/L) MFC = 20 mg/L
Leaves Essential oil Aspergillus Inhibitory effect Deans and inhibits the growth of fungi at different degrees. At the concentration of
and (1.0–10 μL/mL) flavus = 83% at the Svoboda 10 μg/mL and among the fungi tested, Aspergillus niger was the most
stems concentration of (1990) sensitive to OMEO by an inhibitory effect of 89%, followed by Aspergillus
10 μg/mL ochraceus, Aspergillus flavus, Aspergillus parasiticus, and Trichoderma vir­
Aspergillus niger Inhibitory effect
= 89% at the
ide with an inhibitory effect of 84%, 83%, 81%, and 79%, respectively.
concentration of Rus et al. (2015) studied the antifungal activity of OMEO, at concen­
10 μg/mL trations of 0.25 mg/L, 0.5 mg/L, 1 mg/L, 5 mg/L, 10 mg/L, 15 mg/L, 20
Aspergillus Inhibitory effect mg/L, and 0 mg/L for positive control, obtained from the aerial part of
ochraceus = 84% at the
O. majorana using the disc diffusion and microdilution methods, and
concentration of
10 μg/mL Thiophanate-methyl, a commercial agricultural fungicide as negative
Aspergillus Inhibitory effect control. According to them, the growth of Verticillium dahliae was
parasiticus = 81% at the inhibited (diameter of inhibition = 14 mm and MIC = 1 mg/mL) while
concentration of Penicillium aurantiogriseum was inhibited (diameter of inhibition = 10
10 μg/mL
Trichoderma Inhibitory effect
mm and MIC = 1 mg/L). In another work, Hajlaoui et al. (2016) tested
viride = 79% at the the antifungal activity of OMEO (30 mg/mL) against 13 fungal strains: 8
concentration of yeast (Candida albican ATCC 90028, Candida glabrata ATCC 90030,
10 μg/mL Candida parapsilosis ATCC 22019, Candida krusei ATCC 6258, Saccha­
romyces cerevisae, Candida tropicalis, Candida glabrata, Candida albicans,
and Candida Parapsilosis) and 4 Dermatophytic strains (Trichophyton
that dichloromethane extract inhibits the growth of Staphylococcus
violaceum, Trichophyton rubrum, Trichophyton mentagrophytes, and
aureus (30 mm), Escherichia coli (20 mm), Bacillus subtilis (20 mm),
Microsporum canis) using the disc diffusion and microdilution methods
Pseudomonas aeruginosa (15 mm), and Micrococcus flavus (15 mm). The
and Amphotericin B (20 mg/disc) as positive control. The result revealed
diameter of the inhibition zone exhibited by methanol extract varied
that the diameter of inhibition caused by OMEO against human patho­
from 15 to 35 mm, while the most sensitive bacteria were Staphylococcus
genic fungal varied from 11 ± 0 to 28 ± 0 mm and MIC values ranged
aureus (35 mm), Mariniluteicoccus flavus (20 mm), and Pseudomonas
from 0.058 to 0.468 mg/mL.
aeruginosa (20 mm). The aqueous extract exhibited the lowest antibac­
The most sensitive fungus was Candida albicans ATCC 90028
terial activity against the tested bacteria. Another previous research
(Diameter inhibition = 28 ± 0 mm and MIC = 0.058 mg/mL), Candida
reported a great antibacterial activity of ethanol extract (at 5,10, and 20
glabrata ATCC 90030, and Trichophyton mentagrophytes (15.33 ± 0.58
mg) of O. majorana against Gram-positive and Gram-negative bacteria
mm and MIC = 0.117 mg/mL, respectively), and the less sensitive one
(Choi and Rhim, 2008). This study indicated that O. majorana extract
were Saccharomyces cerevisae (11 ± 00 mm, MIC = 0.468 mg/mL) and
inhibits the growth of all bacteria in a concentration-dependent manner,
Trichophyton rubrum (11 ± 00 mm, MIC = 0.234 mg/mL).
while 20 mg/mL exhibited a significant antibacterial effect against all
Della Pepa et al. (2019) evaluated the antifungal activity of OMEO at
strains tested compared to other concentrations with diameter of inhi­
the concentrations of 2000, 1000, and 500 ppm against Botrytis cinerea,
bition zones ranged from 29.5 ± 0.35 to 32.0 ± 0.50 mm.
Penicillium expansum, Aspergillus niger, and Monilinia fructicola, using
The strong antibacterial action of EOs depends on their chemical
azoxystrobin (80 μL.100 mL− 1) as positive control and Tween 20 (0.2%)
composition. Some molecules such as thymol and carvacrol containing
as negative control. The results showed that OMEO inhibits the growth
in EOs have been known as effective antibacterial agents. Moreover, the
of Monilinia fructicola, Penicillium expansum, and Aspergillus niger at a
complex nature of EOs, with several compounds, made these substances
concentration of 2000 ppm. The results of the Fungal Spore Germination
difficult for bacterial strains to develop the resistance against them. In
Assay showed that OMEO inhibits significantly the fungal spore germi­
fact, even the effect of minor elements is not negligible, because they can
nation of Monilinia fructicola in a dose-dependent manner. In fact, the
act synergistically on the antibacterial action.
spore germination was reduced from 93% to 76% by increasing the
The mechanisms of action of essential oils have been demonstrated.
concentration of OMEO from 15 to 45 μL/mL.
The most important action was attributed to their hydrophobicity
Waller et al. (2016) have evaluated the anti-Sporothrix sp. of OMEO
criteria. In fact, The EOs have been known to affect the bacterial
(72–2.25 mg/mL) at dose of 100 μL using microdilution technique with
membrane by increasing its permeability, and as a conclusion, they
Itraconazole as positive control. As a result, all fungi strains (Sporothrix
cause the leakage of vital cell contents (Bouyahya et al., 2019). In
brasiliensis derived from humans, dogs, cats, and Sporothrix schenckii)
addition, other mechanisms of action have been reported such as en­
were sensitive to OMEO action with MIC values varied between ≤ 2.25
zymes inhibitory, DNA fragmentation, and quorum sensing inhibition
and 9 mg/mL. This study listed the fungistatic effect observed at con­
(Bouyahya et al., 2017). Indeed, carvacrol and thymol, the two major
centrations from 2.25 to 9 mg/mL and fungicidal effect occurred be­
compounds of OMEO, showed an important capacity to inhibit Escher­
tween 2.25 and 18 mg/mL. More interesting, Waller et al. (2019) have
ichia coli. This antibacterial action of these compounds was attributed to
studied in vivo the protective effect of OMEO in the cutaneous sporo­
the permeabilization and depolarization of plasma membrane (Xu et al.,
trichosis, caused by Sporothrix brasiliensis, by oral daily treatment of
2008).
control (saline solution), itraconazole (10 mg/kg) and marjoram
essential oil (80 mg/kg) for 30 days. This study showed the antifungal
3.6.2. Antifungal action mechanism
and the systemic protective effects of OMEO by decreasing the fungal
The antifungal activities were reported in the literature indicating
burden in the systemic organs compared to control and itraconazole
high efficacy of EOs and extracts obtained from different parts of
group. The authors suggested OMEO as an alternative treatment of
O. majorana against many fungal strains (Deans and Svoboda, 1990;
cutaneous sporotrichosis.

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A. Bouyahya et al. Journal of Ethnopharmacology 265 (2021) 113318

Table 5
Antioxidant activity of O. majorana.
Part used Extracts Used methods Key results References

Seeds Ethanolic extract DPPH assay Percent Activity = 91.63% Dhull et al. (2016)
ABTS assay Percent Activity = 76.96%
Reducing power assay 7.78 mg AAE/g DWB
Hydroxyl free radical scavenging activity Percent Activity = 5.12%
(HFRSA)
Methanol extract DPPH assay Percent Activity = 91.89%
ABTS assay Percent Activity = 90.79%
Reducing power assay 8.69 mg AAE/g DWB
HFRSA Percent Activity = 0.89%
Acetone extract DPPH assay Percent Activity = 91.03%
ABTS assay Percent Activity = 71.11%
Reducing power assay 6.13 mg AAE/g DWB
HFRSA Percent Activity = 6.13%
Chloroform extract DPPH assay Percent Activity = 84.87%
ABTS assay Percent Activity = 0.76%
Reducing power assay 1.77 mg AAE/g DWB
HFRSA Percent Activity = 0.20%
Leaves Supercritical fluid extractor DPPH assay EC50 = 229.84 mg/mL García-Risco et al.
ABTS assay TEAC = 0.642 mmol/g (2017)
Ultrasonic assisted extraction (ethanol) DPPH assay EC50 = 22.17 mg/mL
ABTS assay TEAC = 0.967 mmol/g
Ultrasonic assisted extraction (ethanol: DPPH assay EC50 = 0.967 mg/mL
water) ABTS assay TEAC = 1550 mmol/g
Aerial parts Essential oil DPPH assay IC50 = 62.66 ± 2.08 μg/mL Hajlaoui et al., 2016
Superoxide quenching activity IC50 = 1.66 ± 0.76 μg/mL
Reducing power IC50 = 2.5 ± 0.28 μg/mL
B-carotens IC50 = 12.83 ± 1.04 μg/mL
Methanolic extract FRAP assay 18.56 g Trolox/100 g DW Hossain et al. (2011)
Aerial parts Essential oil DPPH IC50 = 0.3 ± 0.01 mg/mL Khadhri et al. (2019)
Iron chelating IC50 = 23 ± 0.2 mg/mL
Reducing power IC50 = 0.4 ± 0.01 mg/mL
Ethanolic extract DPPH IC50 = 11.5 ± 0.3 mg/mL
Iron chelating IC50 = 67.2 ± 0.5 mg/mL
Reducing power IC50 = 5.6 ± 0.02 mg/mL
Leaves Aqueous lyophilized extract DPPH IC50 = 12.34 ± 0.1 mg/mL Makrane et al. (2018)
Leaves Methanolic extract DPPH IC50 = 0.0013 ± 0.0001 mg/mL Roby et al. (2013)
Ethanolic extract DPPH IC50 = 0.0025 ± 0.0005 mg/mL
Diethyl ether extract DPPH IC50 = 0.0020 ± 0.0003 mg/mL
Hexane extract DPPH IC50 = 0.0035 ± 0.0009 mg/mL
Methanol extract Superoxide anion radical IC50 = 1.44 ± 0.16 μg/mL Jin Jun et al. (2001)
Leaves Ethanolic extract DPPH assay IC50 = 0.125 mg/mL Abdel-Massih et al.
(2010)
Essential oil DPPH assay Percent Inhibition = 87.9% Baj et al. (2018)
1
Ultrasound-assisted extraction (ethanol/ ABTS assay TEAC = 1.52 ± 0.04 mmol TE g− dry Arranz et al. (2019)
water) extract
1
Pressurized liquid extraction (ethanol/ ABTS assay TEAC = 1.81 ± 0.02 mmol TE g− dry
water) extract
Aerial Essential oil DPPH assay IC50 = 418.80 ± 37.65 μg/mL Salha et al. (2017)
material
Stems Ethanolic extract DPPH assay IC50 = 21.05 μg/mL Vasudeva et al. (2014)
Hydrogen peroxide scavenging activity IC50 = 492.8 μg/mL
Ferric reducing power activity Absorbance = 0.527 ± 0.14
Metal chelating complex assay IC50 = 156.9 μg/mL
Roots Ethanolic extract DPPH assay IC50 = 84.98 μg/mL
Hydrogen peroxide scavenging activity IC50 = 477.6 μg/mL
Ferric reducing power activity Absorbance = 0.747 ± 0.23
Metal chelating complex assay IC50 = 141.79 μg/mL
Leaves Ethanolic extract DPPH assay IC50 = 395.91 ± 0.25 μg/mL Serafini et al. (2012)
ABTS assay AA = 227.03 ± 2.12 μmol trolox g− 1
β-carotene/linoleic acid AA = 82.60 ± 0.27%
FRAP AA = 217.27 ± 0,84 μmol Fe+2 g− 1
Essential oil DPPH assay IC50 = 341.0 μg/mL Schmidt et al. (2008)
Hydroxyl radicals scavenging (OH•) IC50 = 0.11 μg/mL
Areal parts Essential oil DPPH assay IC50 = 1.85 ± 0.02 μg/mL Olfa et al. (2016)
Leaves Methanolic extract DPPH assay IC50 = 12.9 μg/mL Mossa et al. (2013)
ABTS assay IC50 = 9.35 μg/mL
Reducing power IC50 = 21.16 μg/mL
HFRSA IC50 = 9 μg/mL
Leaves Methanolic extract FRAP 17.08 g Trolox/100 g DW Hossain et al. (2011)
Leaves Essential oil DPPH assay IC50 > 250 μg/mL Guerra-Boone et al.
(2015)
Aerial parts Methanolic extract DPPH assay IC50 = 102.6 μg/mL Al-Fatimi (2018)
Aerial parts Ethanolic extract DPPH assay IC50 = 2.77 μg/mL Erenler et al. (2016)
ABTS assay High activity
(continued on next page)

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A. Bouyahya et al. Journal of Ethnopharmacology 265 (2021) 113318

Table 5 (continued )
Part used Extracts Used methods Key results References

FRAP assay IC50 = 0.65 μg/mL

Aerial parts Essential oil DPPH assay IC50 = 170 ± 0.3 μg/mL Erdogan and Ozkan
β-Carotene/linoleic acid assay IC50 = 40.0 ± 0.2 μg/mL (2017)
Leaves Hydro-ethanolic extract DPPH assay IC50 = 37.42 mg/mL Deuschle et al. (2018)
FRAP assay High activity
Leaves Essential oil DPPH assay IC50 = 16.83 μg/mL Chaves et al. (2019)
Leaves Aqueous extract Total scavenging capacity TSC = 100% at the concentration of 0.008 Vagi et al. (2005)
(w/w)
DPPH assay Important antioxidant activity
Ethanol extract, Supercritical CO2 Rancimat method Strong antioxidant activity
extraction
Essential oil DPPH assay Percent Inhibition = 87.61% Abdalla and Hendi
(2014b)

Other studies have focused on the antifungal activity of crude ex­ power, β-carotens, and DPPH assays, respectively. Guerra-Boone et al.
tracts of O. majorana. Indeed, Vagi et al. (2005) studied the antifungal (2015), Salha et al. (2017), and Chaves et al. (2019) have reported an
activity of ethanol extract (2.5% w/v) and supercritical fluid extract anti-DPPH effect of OMEO with IC50 values of >250, 418.80, and 16.83
(0.05, 0.15, 0.25, 0.4, and 0.5% w/v) by daily measurement of the radial μg/mL, respectively.
growth on PDA (potato dextrose agar) plates against Aspergillus niger, In the evaluation of the antioxidant activity of O. majorana EO and its
Trichoderma viride, and Penicillium cyclopium. The results showed that ethanolic extract conducted by Khadhri et al. (2019) using three
the fungi strains tested were more sensitive to the supercritical fluid methods, namely radical scavenging DPPH, iron chelating power, and
extract than the ethanolic extract. In fact, at 0.4% (w/v), the super­ the reducing power assay. This study reports that OMEO, from aerial
critical fluid extract caused an inhibitory effect of 100% against the part, has a good anti-DPPH effect with IC50 = 0.3 mg/mL, an important
growth of Aspergillus niger and Penicillium cyclopium, while its 0.5% reduction power and iron chelating activity with IC50 = 0.4 mg/mL and
(w/v) gave an inhibitory effect of 100% against Trichoderma viride, while IC50 = 23 mg/mL. The ethanol extract showed also an antioxidant effect
a slight inhibitory effect (19.3% inhibition) was found in the presence of with anti-DPPH IC50 value of 11.5 mg/mL, for iron chelating IC50 = 67.2
the ethanolic extract at a concentration of 2.5%. In another work, mg/mL, and IC50 = 5.6 mg/mL for the reducing power.
antifungal activity of the petroleum ether and methanol-water extracts In addition, several scientists have reported the antioxidant activity
at concentrations of 12.5%, 25%, and 50% was determined by agar-well of the crude extract of O. majorana. Roby et al. (2013) examined the
diffusion methods with dimethyl sulphoroxide (DMSO) as negative antioxidant activity of methanol, ethanol, diethyl ether, and hexane
control and standard antibiotics (Nystatine) as positive control. The extracts using DPPH free radical scavenging activity assay. According to
results revealed that the extracts at all concentrations (12.5%, 25%, and this study the interesting IC50 values were in the order of 0.0013 mg/mL,
50%) were not active against fungi organisms (Candida albicans and 0.0020 mg/mL, 0.0025 mg/mL, and 0.0035 mg/mL for methanolic,
Aspergillus niger) (Adam and Ahmed, 2014). diethyl ether, ethanolic, and hexane extracts, respectively. In another
work, Dhull et al. (2016) studied the antioxidant effect of ethanol,
3.6.3. Antioxidant activity methanol, acetone, and chloroform extracts of O. majorana leaves using
Several studies have evaluated the antioxidant activity of EOs and the DPPH, ABTS, reducing power, and hydroxyl free radical scavenging
extracts obtained from different parts of O. majorana (Schmidt et al., activity (HFRSA) methods. The results showed that all extracts have an
2008; Serafini et al., 2012; Mossa et al., 2013; Roby et al., 2013; Abdalla important anti-DPPH activity with a percent activity of 91.89%,
and Hendi, 2014a; Vasudeva et al., 2014; Guerra-Boone et al., 2015; 91.63%, 91.03%, and 84.87% caused by methanol, ethanolic, acetone,
Hajlaoui et al., 2016; Dhull et al., 2016; García-Risco et al., 2017; and chloroform extracts, respectively.
Erdogan and Ozkan, 2017; Salha et al., 2017; Chaves et al., 2019; Another research work reported the antioxidant effect of ethanol
Khadhri et al., 2019; Arranz et al., 2019). All of these studies were extract of O. majorana obtained from leaves and roots using the DPPH
carried out by DPPH, FRAP, Beta-carotene, ABTS, HFRSA, superoxide assay, hydrogen peroxide scavenging activity, ferric reducing power
quenching activity, and TAC assays. activity, and metal chelating complex assay (Vasudeva et al., 2014). As a
Table 5 summarizes all studies that have evaluated the antioxidant result, the ethanol extract from leaves exerted a good antioxidant effect
activity of O. majorana. It presents the part used, the type of extract, the in which the IC50 values were 21.05 μg/mL for DPPH, 156.9 μg/mL for
methods used, and the main results. Indeed, Abdalla and Hendi (2014a), metal chelating complex, and 477.6 μg/mL for hydrogen peroxide
studied the anti-radical activity of OMEO using DPPH as free radical. scavenging. While the ferric reducing power was expressed as absor­
The results showed that the percentage of inhibition caused by OMEO bance with a value equal to 0.527. For the ethanol extract of the roots,
was 87.61%. Schmidt et al. (2008) have reported the antioxidant effect the results showed that the extract inhibits the DPPH with IC50 = 84.98
of OMEO using DPPH and hydroxyl radicals scavenging (OH•) assay. μg/mL, IC50 = 477.6 μg/mL for hydrogen peroxide scavenging, and IC50
This study demonstrated that the IC50 of OMEO was 0.11 and 341.0 = 141.79 μg/mL for Metal chelating complex, while the reduction power
μg/mL, using (OH•) and DPPH, respectively. The results of the study absorbance was 0.747 ± 0.23. The antioxidant propriety of ethanol
reported by Erdogan and Ozkan (2017) showed that the inhibition of extract of O. majorana was also studied by Serafini et al. (2012) using
β-Carotene bleaching activity of OMEO obtained from the aerial parts DPPH, ABTS, β-carotene/linoleic acid, and FRAP methods. This extract
was 40.0 ± 0.2%, while the concentration for 50% inhibition of DPPH possesses an anti-DPPH activity with IC50 = 395.91 ± 0.25 μg/mL. The
(IC50) was 170 μg/mL. The anti-DPPH effect of the aerial part of OMEO ABTS analysis indicates the antioxidant activity in the ethanol extract of
was reported by Salha et al. (2017) which showed that the IC50 = 227.03 ± 2.12 μmol TEAC g/L. For the ferric reducing antioxidant
418.80 ± 37.65 μg/mL. In another work, Hajlaoui et al. (2016) inves­ power, the extract showed activity of 217.27 ± 0.84 μmol Fe2+ g/L of
tigated the antioxidant effect of OMEO using DPPH radical scavenging the plant.
assay, superoxide anion radical-scavenging activity, reducing power Mossa et al. (2013) used DPPH, ABTS, reducing power, and hydroxyl
method, and β-Carotene-linoleic acid model system. The results of this radicals scavenging methods to study the antioxidant activity of meth­
study indicate that the IC50 values were 1.66 μg/mL, 2.5 μg/mL, 12.83 anol extract of O. majorana. The results showed a great antioxidant
μg/mL, and 62.66 μg/mL for superoxide quenching activity, reducing property of this extract with IC50 values of 9 μg/mL, 9.35 μg/mL, 12.9

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A. Bouyahya et al. Journal of Ethnopharmacology 265 (2021) 113318

μg/mL, and 21.16 μg/mL for hydroxyl radicals scavenging, ABTS assay, study, El-Akhal et al. (2014), have tested the larvicidal effect of
DPPH assay, and Reducing power, respectively. O. majorana EO against Culex pipiens and West Nile virus vector. The
The antioxidant activity of O. majorana extract has been proven by results showed that LC50 and LC90 values were 258.71 and 580.49 mg/L,
García-Risco et al. (2017). The antioxidant effect was determined by respectively (El-Akhal et al., 2014).
different methods: the DPPH radical assay, expressed as EC50 value and The literature reported also that O. majorana EO presented an
the ABTS radical method expressed as TEAC values (mmol TE/g extract). effective effect against the beetle Tribolium castaneum (Sharma et al.,
Both methods demonstrated a good antioxidant activity with the EC50 2016). The treatment with EO at 1000 and 500 ppm exhibited a
value of 13.74 mg/mL and TEAC value of 1550 mmol TE/g extract for of maximum mortality. Moreover, the larvicidal activity of O. majorana
ultrasonic assisted extraction (ethanol:water), while for ultrasonic was tested against the third instar larvae of Aedes Aegypti, Anopheles
assisted extraction (ethanol), the EC50 value was equal to 22.17 mg/mL stephensi, and Culex quinquefasciatus, the findings of this work showed
and TEAC value was equal to 0.967 mmol TE/g extract. For supercritical that the lethal concentration (LC50) values were 519.07, 500.08, and
fluid extract, the EC50 was equal to 229.84 mg/mL and TEAC value was 527.651 ppm, respectively (Sefeer and Elumalai, 2018). In another
equal to 0.642 mmol TE/g extract. study, the larvicidal activity showed that O. majorana EO caused high
Another experiment conducted by Arranz et al. (2019) indicates the mortality in larvae of the third stage of A. aegypti larvae with LC50 =
antioxidant activity of ultrasound-assisted extraction (ethanol/water) 15.696 μg/mL after 48 h (Chaves et al., 2019). Also, Salaheddine et al.
and pressurized liquid extraction (ethanol/water) of O. majorana leaves. (2013) tested the insecticidal activity of essential oil from O. majorana
Indeed, TEAC value was 1.52 ± 0.04 mmol TE g/L dry extract and 1.81 against adults of Tribolium castaneum and larvae of Plodia interpunctella.
± 0.02 mmol TE g/L dry extract for ultrasound-assisted extraction The insecticidal activity of leaves oil against T. castaneum adults was
(ethanol/water) and pressurized liquid extraction (ethanol/water), higher (LC50 = 73.75 μl/L air). However, Plodia interpunctella larvae
respectively. were exposed to higher doses (LC50 = 329.5 μl/L air). For flower oil, the
insecticidal activity obtained was interesting (LC50 = 76.5 μL/L air for
3.6.4. Antiparasitic and insecticide activities T. castaneum and LC50 = 329.55 μl/L air for Plodia interpunctella) (Sal­
Mosquitoes have always been considered a source of nuisance for aheddine et al., 2013).
humans, mainly due to the fact that they can be vectors of diseases. The
use of chemical insecticides is currently the most practiced technique to 3.6.5. Antidiabetic activity
fight against insects but can cause various environmental problems Several studies have reported the in vitro and in vivo antidiabetic
(Barbouche et al., 2001; Sinegre et al., 1977). To ensure better inter­ effect of O. majorana (Table 7). The leaf methanolic extract containing
vention, the implementation of new mosquito control alternatives is five hydroxyflavonids was tested against α-Glucosidase in an in vitro
more encouraged. In this context, the plants are a source of natural study (Kawabata et al., 2003). The results revealed that hydrox­
substances that have great potential for application against insects and yapigenin exhibit the potent inhibitory effect on α-Glucosidase (IC50 =
other pests. Indeed, the insecticidal activity of O. majorana EOs was 12 μM). Using, alloxan induced diabetic rats model, Oaman and Abbas
executed against various insects which showed that EOs displayed (2010) showed that the aqueous extract from O. majorana leaves
considerable activity (Table 6). For example, El-Akhal et al. (2016), decreased importantly blood glucose, amylase, insulin, testosterone, and
evaluated the larvicidal activity of essential oils from O. majorana cholesterol levels in diabetic rats. On the other hand, the hypoglycemic
against the larvae of the malaria vector Anopheles labranchiae (Diptera: effects of O. majorana leaf extracts (volatile oil, methanol, and aqueous
Culicidae). The results showed that O. majorana essential oil exhibited a extracts) has been tested in vivo using nicotinamide and streptozotocin
larvicidal effect, with the lethal concentration (LC50 = 107.13 μg/mL (STZ) diabetic rat model (Pimple et al., 2012). The study showed that
and LC90 = 365.90 μg/mL) after 24 h (El-Akhal et al., 2016). In another oral administration of extracts at 100, 200, and 400 mg/kg have
significantly reduced blood glucose and increased hemoglobin A1c
(HbA1c) level and blood insulin in diabetic rats in the case of volatile
Table 6 and methanol extracts. However, the aqueous extract reduced impor­
Antiparasitic activity of O. majorana. tantly blood glucose and increased hemoglobin A1c (HbA1c) level, but
Part Extracts Tested strains Key results References without affecting basal plasma insulin concentrations in diabetic rats
used (Pimple et al., 2012).
Leaves Essential Aedes aegypti After 48 h, LC50 Chaves et al. Using the same experimental model, Perez Gutierrez (2012) revealed
oil = 15.696 μg/mL (2019) that O. majorana leaf methanolic extract administrated at 200 mg/kg
Vegetal Essential Anopheles CL50 = 107.13 El-Akhal et al. reduces Advanced Glycation End products (AGEs) and TBA-reactive
matte oil labranchiae μg/mL (2014)
substance level and decreases the levels of serum protein and hemo­
(Diptera:
Culicidae) globin glycosylated. The results showed also a reduction of tail tendon
Essential Culex pipiens CL50 = 258.71 collagen and the levels of collagen-linked fluorescence, and an increase
oil μg/L of pepsin soluble collagen in diabetic rats (Perez Gutierrez, 2012).
Essential Aedes aegypti LC50 = 519.07 Sefeer and Moreover, the administration of food containing O. majorana powder
oil pmm Elumalai
Essential Anopheles stephensi LC50 = 500.08 (2018)
(from leaves) in STZ diabetic rats induced a significant decrease in the
oil ppm serum level of glucose (p < 0.05) and MDA level (p = 0.04) versus un­
Essential Culex LC50 = 527.651 treated diabetics, as well as protection of the renal tissue through
oil quinquefasciatus ppm attenuation of glomerular expansion and oxidative stress (Moghaddam
Essential Tribolium Maximum Sharma et al.
et al., 2013). Recently, Tripathy et al. (2018) reported that O. majorana
oil castenium L. mortality with (2016)
1000 and 500 ethanolic extract administrated at different doses (100, 200, and 400
ppm treatments mg/kg), reduced blood glucose and enhanced serum insulin levels in
Leaves Essential Tribolium LC50 = 73.75 Salaheddine STZ diabetic rats. This extract exhibited also a significant decrease (p <
oil castaneum μL/L air et al. (2013) 0.05, P < 0.01) of total cholesterol, triglycerides, LDL, and VLDL levels
Larvae of Plodia LC50 = 382.25
interpunctella μL/L air
(156.5 ± 2.05, 88.15 ± 1.93, 90.05 ± 1.25, 21.51 ± 0.62 mg/dL,
Flowers Essential Tribolium LC50 = 76 μL/L respectively) versus untreated diabetics (184.6 ± 0.91, 98 ± 2.86, 109.7
oil castaneum air ± 1.62, and 18.89 ± 0.56 mg/dL, respectively) and a significant increase
Larvae of Plodia LC50 = 329.5 (p < 0.05) of HDL levels (43.5 ± 0.11 mg/dL) versus untreated diabetics
interpunctella μL/L air
(20.18 ± 0.37 mg/dL) (Tripathy et al., 2018).

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A. Bouyahya et al. Journal of Ethnopharmacology 265 (2021) 113318

Table 7
Antidiabetic activity of O. majorana.
Part Extract tested/compound Dose Model Keys results References
used

Leaves Hydrodistilled volatile oil 100, 200, and 400 Nicotinamide + Significantly reduced blood glucose and increased Pimple et al.
Methanolic extract mg/kg body weight Streptozotocin (STZ) haemoglobin A1c (HbA1c) level and blood insulin in (2012)
Aqueous extract diabetic rats diabetic rats
Leaves Ethanolic extract 100, 200, and 400 Streptozotocin (STZ) Significantly reduced blood glucose Tripathy et al.
mg/kg body weight diabetic rats Enhanced serum insulin levels of the treated diabetic (2018)
rats
Significantly decreased total cholesterol, triglycerides,
LDL, and VLDL levels
Increased HDL levels in diabetic rats
Leaves Methanolic extract 200 mg/kg body Streptozotocin (STZ) Significantly reduced AGE and TBA-reactive substance Perez Gutierrez
weight diabetic rats level (2012)
Decreased the levels of serum protein and hemoglobin
glycosylated
Significantly reduced tail tendon collagen and the levels
of collagen-linked fluorescence
Increased pepsin soluble collagen in diabetic rats.
Leaves Powder Food containing OM Streptozotocin (STZ) Significantly reduced the serum level of glucose and Moghaddam et al.
powder diabetic rats MDA level (2013)
Protected the renal tissue via attenuation of oxidative
stress
Leaves Aqueous extract 20 and 40 mg/kg Alloxan induced diabetic rats Significantly decreased blood glucose, amylase, insulin, Oaman and Abbas
body weight testosterone, and cholesterol levels in diabetic rats (2010)
Leaves Methanolic extract (Five Nd α-Glucosidase IC50 = 12 μM for 6-hydroxyapigenin Kawabata et al.
hydroxyflavonids) (2003)

The antidiabetic effect of O. majorana extract is due to the presence phosphoenolpyruvate carboxykinase (PEPCK) (van Son et al., 2011).
of phenolic acid and flavonoid compounds such as rosmarinic acid, Additionally, ferulic acid may also decrease glucose levels by mitigating
ferulic, apigenin, hesperetin, luteolin, arbutin, quercetin, and catechin. pancreatic damage, increasing insulin release, and decreasing hepatic
Indeed, some studies revealed that ferulic acid exhibits the antidiabetic glycogenolysis (Azay-Milhau et al., 2013). On the other hand, apigenin
effects via several mechanisms such as increasing the plasma insulin ameliorates importantly glucose homeostasis by decreasing the levels of
levels, stimulating hepatic glycogen synthesis, increasing glucokinase blood glucose, serum lipid and insulin resistance index, and improving
activity, down-regulating glucose-6-phosphatase (G6pase) and impaired glucose tolerance (Ren et al., 2016). Moreover, it ameliorates

Table 8
Anticancer activity of O. majorana.
Part used Extracts Cell lines Key results References

Leaves Supercritical fluid extractor Pancreatic human tumor-derived cell line miapaca-2 IC50 > 100 μg/mL García-Risco et al.
Ultrasonic assisted extraction Pancreatic human tumor-derived cell line miapaca-2 IC50 > 100 μg/mL (2017)
(ethanol)
Ultrasonic assisted extraction Pancreatic human tumor-derived cell line miapaca-2 IC50 > 100 μg/mL
(ethanol:water)
Aerial Essential oil Human tumor cell lines, Hep2 CC50 = 85.63 ± 2.38 mg/mL Hajlaoui et al., 2016
parts Human tumor cell lines, HT29 CC50 = 13.73 ± 1.31 mg/mL
Continuous cell lineage control (Vero) CC50 = 70.13 ± 1.72 mg/mL
Aerial Aqueous extract Human breast cell line MDA-MB-231 IC50 = 69.18 ± 3.10 μg/mL Makrane et al. (2018)
parts Human colon cell line HT-29 IC50 = 177.82 ± 4.07 μg/mL
Depleted aqueous extract Human breast cell line MDA-MB-231 IC50 = 87.09 ± 2.39 μg/mL
Human colon cell line HT-29 IC50 = 158.48 ± 1.23 μg/mL
Petroleum ether extract Human breast cell line MDA-MB-231 IC50 = 43.65 ± 2.63 μg/mL
Human colon cell line HT-29 IC50 = 63.09 ± 3.65 μg/mL
Dichloromethane extract Human breast cell line MDA-MB-231 IC50 = 53.70 ± 7.94 μg/mL
Human colon cell line HT-29 IC50 = 125.89 ± 7.86 μg/mL
Ethyl acetate extract Human breast cell line MDA-MB-231 IC50 = 30.90 ± 1.39 μg/mL
Human colon cell line HT-29 IC50 = 50.11 ± 1.44 μg/mL
Methanolic extract Human breast cell line MDA-MB-231 IC50 = 48.97 ± 7.97 μg/mL
Human colon cell line HT-29 IC50 = 141.25 ± 1.16 μg/mL
Leaves Lyophilized aqueous extract Fibrosarcoma cancer cell line HT-1080 Not cytotoxic Rao et al. (2014)
Ethanolic extract Fibrosarcoma cancer cell line HT-1080 IC50 = 110 μg/mL
Methanolic extract Fibrosarcoma cancer cell line HT-1080 Not cytotoxic
Leaves Ethanolic extract Human breast cancer cells MDA-MB-231 IC50 = 400 mg/mL Al Dhaheri et al. (2013)
Leaves Ethanolic extract Human lymphoblastic leukemia cell line Jurkat IC50 = 5 mg/mL Abdel-Massih et al.
(2010)
Leaves Ethanolic extract Human colorectal cancer cell lines HT-29human IC50 = 342 μg/mL Benhalilou et al. (2019)
colorectal cancer cell lines Caco-2 IC50 = 296 μg/mL
Leaves Methanolic extract Human colon cancer HT-29 cell line Inhibited the growth of HT-29 cells by Al Tamimi (2015)
effecting the cells viability
Leaves Aqueous and ethanol extracts Human hepatocellular carcinoma (HepG2) cell line Highly significant inhibitory effect on HepG2 Fathy et al. (2016)
cell proliferation
Leaves Aqueous extracts HeLa, MCF-7, and Jurkat cancer cells Hight antiproliferative inhibition Elansary and Mahmoud
(2015)

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A. Bouyahya et al. Journal of Ethnopharmacology 265 (2021) 113318

also the vascular endothelial dysfunction, increases insulin-mediated extracts obtained by supercritical fluid extractor were evaluated for
NO production and inhibits NF-kB-mediated inflammatory response in their cytotoxic effects against pancreatic human tumor-derived cell line
endothelial cells (Jung et al., 2016). In addition, by using Western blot miapaca-2. The results showed that all extracts revealed moderate
analysis, Wang et al. (2017) revealed that the pretreatment with api­ cytotoxic effects with IC50 > 100 μg/mL (García-Risco et al., 2017).
genin effectively reduces ROS levels and restores the cell apoptosis of Hajlaoui et al. (2016) evaluated the antiproliferative effect of
pancreatic β-cells stressed by different concentrations of STZ. Further­ O. majorana EO on two human tumor cell lines (Hep2 and HT29) and
more, Esmaeili et al. (2009) showed that this compound increases in­ Vero cell line. The results showed that O. majorana EO inhibited the
sulin secretion and protects pancreatic β-cells from oxidative stress proliferation of Hep2, HT29, and Vero with IC50 values of 85.63 ± 2.38,
induced by STZ. 13.73 ± 1.31, and 70.13 ± 1.72 mg/mL, respectively. In another study,
different extracts (aqueous, petroleum ether, dichloromethane, ethyl­
3.6.6. Anticancer activity acetate, and methanolic extracts) prepared from the same part of
Origanum majorana has also been studied for its anticancer proper­ O. majorana were investigated for their anticancer effects on human
ties. Indeed, several in vitro investigations based on cell culture tests breast cell line MDA-MB-231 and human colon cell line HT-29 (Makrane
showed that O. majorana extracts and essential oils exhibit anti­ et al., 2018). The results demonstrated that O. majorana ethyl acetate
proliferative effects against different cancer cell lines (Abdel-Massih extract exhibited the best antiproliferative effect on both MDA-MB-231
et al., 2010; Al Dhaheri et al., 2013; Rao et al., 2014; Hajlaoui et al., and HT-29 cell lines at IC50 = 30.90 ± 1.39 and IC50 = 50.11 ± 1.44
2016; García-Risco et al., 2017; Makrane et al., 2018; Benhalilou et al., μg/mL, respectively (Makrane et al., 2018). Methanolic, ethanolic, and
2019) (Table 8). lyophilized aqueous extracts of O. majorana leaves have been tested by
Volatile compounds extracted, and ethanol and water alcoholic Rao et al. (2014) on fibrosarcoma cancer cell line HT-1080. In this study,

Table 9
Other pharmacological activities of O. majorana.
Activities Use part Extracts Experimental approach Key results References

Nephroprotective Flowers Ethanolic extract Cisplatin induced the nephrotoxicity in rats Reduced the creatinine, urea, uric acid, and BUN Soliman et al.
effect (500 mg/kg body levels by 57.78%, 55.39%, 42.87%, and 37.67%, (2016).
weight) for 14 days respectively
Significantly potentiated the renal antioxidant
molecules such as GSH, SOD, and CAT by 38.56%,
35.42%, and 48.42%, respectively
Significantly lowered the MDA and NO levels by
107.74% and 73.02%, respectively
Anti-inflammatory Aerial Ethanolic extract Carrageenan-induced paw oedema in rats Significantly decreased the paw oedema rate at a Seoudi et al.
effect parts (0.25 and 0.5 g/kg dose of 0.25 and 0.5 g/kg body weight in (2009)
body weight) comparison to control non-treated group and
indomethacin (standard drug) group
Analgesic effect Aerial Ethanolic extract Hot plate method Increased the reaction time (12.20 s at zero time) for Seoudi et al.
parts (0.25 and 0.5 g/kg the dose of 0.25 g/kg body weight to 15.60 s at 30 (2009)
body weight) min post administration
Anti-pyretic Effect Aerial Ethanolic extract Brewer’s yeast-induced pyrexia in rats Significantly decreased the induced rise in rectal Seoudi et al.
parts (0.25 and 0.5 g/kg temperature compared to control (2009)
body weight)
Hepatoprotective Leaves Essential oil (160 L/ Prallethrin induced oxidative stress and Significantly reduced the activities of ALT, AST, and Mossa et al.
effect kg body weight) hepatotoxicity in rat ALP compared with prallethrin group (2013)
Significant reduction in MDA in prallethrin-treated
rats
Improved significantly the activities of CAT, SOD,
and GST in liver compared with control values
Antimutagenic Leaves Aqueous extract Vicia faba root meristematic cells exposed to Showed chromosomal aberrations at 50, 100, and Qari (2008)
effect (50, 100, and 200 250 and 350 μg/mL of sodium azide for 6 h 200 μg/mL
μg/mL) Decreased significantly the number of abnormal
metaphases and the number of aberrations
Decreased the percentage of mitotic indexes
Leaves Oil extract (1.25 Allium cepa root tips mono-sodium glutamate Increased mitotic index Khatab and
μL/mL) (MSG) induced mutagenicity Reduced the chromosomal aberration Elhaddad
(2015)
Antiulcer effect Aerial Ethanol extract Hypothermic restraint stress-, indomethacin- Significantly decreased the incidence of ulcers, Al-Howiriny
parts (250 and 500 mg/ , necrotizing agents induced ulcers and basal basal gastric secretion, and acid output et al. (2009)
kg of body weight) gastric acid secretion using pylorus ligated Significantly lowered the increase in the
Shay rat-model concentration of malondialdehyde (MDA)
Acute toxicity Aerial Ethanolic extract Oral single administration of extract at doses No mortalities in rats following oral administration Seoudi et al.
parts (1, 2, 3, 4, and 5 g/kg body weight) to rats in doses up to 5 g/kg body weight (2009)
Behavioral changes, symptoms of toxicity,
and mortality were observed for 24 h
Aerial Essential oil Oral single administration of origanum No toxicity symptoms occurred Selim et al.
parts majorana oil at doses of 80, 160, and 320 (2013)
mg/kg, respectively to hamsters
Determination of mortality in 14 days
Preventive effects Leaves Aqueous extract Isoproterenol (ISO)-induced haematological Significantly alleviated erythrocytosis, Ramadan et al.
changes and myocardial oxidative stress rats granulocytosis, thrombocytosis, and shortened (2013)
clotting time
Significantly alleviated the increase in relative heart
weight, myocardial oxidative stress, and the leakage
of heart enzymes

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only the ethanolic extract has exhibited a cytotoxic effect of the tested effect was carried out using brewer’s yeast-induced pyrexia in rat
cell line (IC50 = 110 μg/mL). Recently, Benhalilou et al. (2019) showed model. The ethanolic extract of O. majorana administered (0.25 g/kg b.
that O. majorana ethanolic extract had important antiproliferative ef­ wt.) to animals reduced importantly the induced rise in rectal temper­
fects against human colorectal cancer cell lines HT-29 (IC50 = 342 ature compared with the control. Moreover, during 4 h of experience,
μg/mL) and human colorectal cancer cell lines Caco-2 (IC50 = 296 the ethanolic extract (0.5 g/kg b.wt.) decreased also the rectal temper­
μg/mL). ature with a percent throughout time intervals of the experiment (2.8,
The main volatile compounds of OMEO such as carvacrol and thymol 4.3, 5.0, and 5.3%, during 4 h, respectively). This effect was important
demonstrated remarkable anticancer effects with several mechanisms. compared with the control group (Seoudi et al., 2009).
These molecules induced cytotoxicity of cancer lines via numerous ac­ Phenolic compounds of O. majorana extracts exhibit anti-
tions such as cell cycle arrest, apoptosis, autophagy, inhibition of cell inflammatory properties via different actions. The reported studies
migration, and angiogenesis (Bouyahya et al., 2020; Elbe et al., 2020; showed that quercetin (main flavonoid compound) reduces the expres­
Pakdemi˙rli˙ et al., 2020). Flavonoids and phenolic acids identified in sion of TNF-α in mouse model of endotoxemia (Penalva et al., 2017).
O. majorana extracts such as rosmarinic acid, ferulic, apigenin, luteolin, Moreover, this molecule had inhibitory effects of different inflammatory
and quercetin exhibit several mechanisms of action on human cancers mediators such as NO, IL-6, MCP-1, IP-10, RANTES, GM-CSF, G-CSF,
(Kopustinskiene et al., 2020; Khan et al., 2020). These bioactive com­ TNF-α, LIF, LIX, and VEGF. Molecular investigations showed that this
pounds target cancer cell lines on different checkpoints such as targeting inhibition involves the repression of activated transcription 1(STAT1)
cell death, cell cycle arrest, inducing apoptosis, autophagy, telomerase and STAT3 which regulate the transcriptional activity of these mediators
activity, modulating ROS effects, suppressing cancer cell proliferation (Kim and Park, 2016). Moreover, quercetin inhibited the enzymatic
and invasiveness (Khan et al., 2020; Kopustinskiene et al., 2020). activity of pro-inflammatory 12/15-lipoxygenase in lung and liver as
well as their expression. These actions were associated with a decrease
3.6.7. Nephrotoxicity protective effect in transcriptional factor NF-κB activity (Gardi et al., 2015).
Soliman et al. (2016) investigated the anti-nephrotoxicity effect of Phenolic acids such as ferulic acid also showed an anti-inflammatory
O. majorana ethanolic extract. In this study, the authors used cisplatin effect (Cao et al., 2015). This compound was reported to be able to
anticancer drugs to induce the nephrotoxicity in rats and tested thus the decrease IL-6, IL-1β, and TNF-α levels and to inhibit iNOS gene
preventive effect of ethanolic extract of O. majorana. The experiment expression in H2O2-induced cell culture rat vascular smooth muscle cells
divided into two parts: a group treated only with cisplatin at 3 mg/kg of (VSMCs) as well as H2O2-induced iNOS mRNA expression. This inhibi­
body weight), and a group treated with both O. majorana ethanolic tion involves a decrease in p22phox, p47phox, and gp91phox mRNA
extract and cisplatin 500 mg/kg of body weight. After 14 days of expression and a reduction of gp91phox, p47phox, and NOX4 levels in
treatment, cisplatin showed significant (p < 0.05) toxicity revealed by H2O2-induced VSMCs (Cao et al., 2015). Moreover, Doss et al. (2016)
the increase of creatinine, urea, uric acid, blood urea nitrogen, malon­ evaluated the anti-inflammatory activity of ferulic acid against mono­
dialdehyde, and nitric oxide levels (0.727 ± 0.047 mg/dL, 0.727 ± sodium urate crystal-induced inflammation in rats. The results showed a
0.047 g/dL, 2.742 ± 0.165 mg/dL, 116.911 ± 8.654 g/dL, 12.029 ± decrease in paw oedema with a significant reduction in the levels of
0.711 nmol/g protein, and 312.555 ± 19.110 Mmol/g protein, respec­ articular elastase and lysosomal enzymes. Ferulic acid also suppressed
tively) as compared to control group (0.448 ± 0.013 mg/dL, 38.145 ± TNF-α, IL-1β, NLRP3, and caspase-1 expression throughout the inhibi­
1.065 g/dL, 2.148 ± 0.081 mg/dL, 81.718 ± 2.278 g/dL, 5.937 ± 0.595 tion of transcriptional factor NF-kB (Doss et al., 2016). In another
nmol/g protein and 171.602 ± 9.944 Mmol/g protein, respectively). In experimental model, in mice exposed to chronic unpredictable mild
addition, enzymes of antioxidant defense such as glutathione, superox­ stress; ferulic acid decreased several neuro-inflammatory factors such as
ide dismutase, and catalase were repressed (13.621 ± 2.098 mg/g IL-1β and TNF-α in the prefrontal cortex (Liu et al., 2017).
protein, 655.220 ± 87.654 U/g. protein and 5.730 ± 0.776 U/g. protein,
respectively) versus control group (22.719 ± 2.885 mg/g protein, 3.6.9. Hepatoprotective effects
993.255 ± 74.932U/g. protein, and 11.054 ± 0.502 U/g. protein, The hepatoprotection of O. majorana was reported by Mossa et al.
respectively). Importantly, the incorporation of O. majorana ethanolic (2013) (Table 9). In this study, the authors used Prallethrin as a toxic
extract with cisplatin has conversantly modulated the oxidative markers agent for inducing oxidative stress and hepatotoxicity in rat. This
and reduced significantly the nephrotoxicity induced by cisplatin via treatment caused an important decrease in body weight gain (8.13%)
antioxidative effects (scavenging activity) (Soliman et al., 2016) compared to control group (13.53%) and a significant increase in rela­
(Table 9). tive liver weight compared to control (3.95% versus 2.84%). In addition,
the activity of several liver enzymes (aspartate transaminase, alanine
3.6.8. Anti-inflammatory, analgesic, and anti-pyretic effects transaminase, alkaline phosphatase, and serum marker enzymes) was
Ethanolic extract of O. majorana was evaluated by Seoudi et al. significantly increased (p < 0.05). It had also the capacity to reduce
(2009) for its anti-inflammatory, analgesic, and anti-pyretic properties significantly (p < 0.05) the activity of several other enzymes, including
(Table 9). The anti-inflammatory effect was carried out in vivo using superoxide dismutase, catalase (13.90 versus 30.22 μmol/mg protein),
carrageenan-induced paw oedema in rats. It has shown as results that and glutathione-S-transferase (452.62 versus 643.01 μmol/mg protein)
the pretreatment with the ethanol extract of O. majorana at two doses in the liver compared to the control group. Interestingly, the incorpo­
(0.25 and 0.5 g/kg of body weight) remarkably decreased the paw ration of O. majorana EOs (160 μL/kg b.wt.) revealed remarkable pro­
oedema rate in comparison with control (non-treated group). At 0.25 tection of hepatotoxicity induced by prallethrin. Indeed, O. majorana EO
g/kg of body weight, during 4 h, this extract inhibited the induced has established almost all serum marker enzymes and the antioxidant
inflammation by 28.6, 21.3, 16.3, and 12.4%, for each hour, respec­ status and brought all the values near to normal, which indicate that the
tively. However, at 0.5 g/kg of body weight dose, O. majorana ethanol protective effect of this oil was mediated by suppressing oxidative
extract exhibited inhibition of 35.4, 33.3, 28.1, and 30.5%, during 4 h of damage and liver injury in rats (Mossa et al., 2013).
treatment, respectively (Seoudi et al., 2009). Moreover, the hot plate
method was used to study the analgesic effect of the same extract. The 3.6.10. Antimutagenic
authors showed an important increase in reaction time (pain latency) Aqueous and oil extracts from O. majorana were investigated for
when compared to the control (non-treated group and their basal their anti-mutagenic effects (Table 9). The study of Qari (2008) inves­
values). At 0.25 g/kg of body weight treatment with O. majorana etha­ tigated the in vitro anti-mutagenic effect of O. majorana extract on the
nolic extract, the reaction time for the dose was 12.20 s at zero time and meristematic root cells of Vicia faba. The roots of this later were treated
increased to 15.60 s at 30 min (Seoudi et al., 2009). The anti-pyretic with sodium azide at 250 and 350 μg/mL for 6 h, and then the

26
A. Bouyahya et al. Journal of Ethnopharmacology 265 (2021) 113318

administration of O. majorana aqueous extracts was performed at 50, 3.8. Clinical evaluation
100, and 200 μg/mL for 20 h, prior to sodium azide treatment (Qari,
2008). The results showed that the treatment of Vicia faba root meri­ Clinical investigations of some O. majorana bioactive compounds
stematic cells induced important chromosomal aberrations with including quercetin have been showed. Indeed, this molecule demon­
increasing concentrations. The total number of aberrations was signifi­ strated important clinical anti-inflammatory effects by inhibiting the
cantly reduced in root tip cells pretreated with O. majorana. However, generation of inflammatory mediators such as cytokines and chemo­
these aberrations were importantly reduced with the incorporation of kines (McAnulty et al., 2008). Moreover, in another study, this molecule
O. majorana extract. It was concluded that O. majorana extracts has an decreased the expression of numerous inflammatory factors and medi­
important antimutagenic potential against sodium azide induced chro­ ators also in athletics (Konrad et al., 2011). It has been showed that
mosomal aberrations in Vicia faba root meristematic cells (Qari, 2008). quercetin decreases also the level of c-reactive protein (CRP), stimulates
In another study, the anti-mutagenic activity of O. majorana was factors of the colonies of granulocytes and macrophages (GM-CSF) and
carried out on Allium cepa root tips mono-sodium glutamate (MSG) the interleukins (IL-10, IL-1β, IL-2, IL-6, and IL-8). In another work on
induced mutagenicity (product which induces several mitodepression athletics (McAnulty et al., 2013) quercetin significantly increased the
chromosomal aberrations such as disturbance, sticky chromosomes, level of IL-8 and CRP after exercise. From a pathological point of view,
bridges, fragments, and other morphological abnormalities like cells quercetin showed its ability to reduce inflammation in patients with
enlargement) (Khatab and Elhaddad, 2015). Importantly, O. majorana sarcoidosis by reducing inflammatory factors, in particular TNF-α/IL-10
oil extract (1.25 μl/mL) inhibited the mutagenic effect induced by MSG and IL-8/IL-10 (Boots et al., 2011). Epicatechin, has also presented an
via the diminishing of mitotic index and reduction of the chromosomal important clinical effectiveness in clinical trials (Dower et al., 2015).
aberration. These findings suggest that O. majorana could be a promising Indeed, it has been showed that pure epicatechin supplementation in
source of antimutagenic and antigenotoxic potential (Khatab and healthy hypertensive women and men in phase III showed significant
Elhaddad, 2015). effect on inflammatory markers or the z-score of inflammation. More­
over, the consumption of gallic acid (15 mg/w/d for seven days)
3.6.11. Protection of gastric mucosal reduced importantly CRP (by 39%) (Ferk et al., 2018).
The ethanolic extract of O. majorana was tested as an antiulcerogenic
agent in vivo using rat model (Al-Howiriny et al., 2009) (Table 9). Ulcers 4. Concluding remarks and future perspectives
were induced using hypothermic restraint stress, indomethacin, and
necrotizing agents (80% ethanol, 25% NaCl, and 0.2M NaOH), while Here, we highlighted a review about O. majorana concerning its
pylorus ligated Shay rat-model was employed to induce basal gastric taxonomy, description, distribution, medicinal use, bioactive com­
acid secretion. The results showed that O. majorana ethanolic extract pounds, pharmacological properties, and toxicological evidences. The
administered orally at 250 and 500 mg/kg b.wt. has significantly (p < data reported that this species is used in traditional medicine to treat
0.05-p<0.001) decreased the incidence of ulcers (13.50 ± 4.08 and different pathologies including microbial infections, inflammation, and
11.66 ± 1.50 at doses of 250 and 500 mg/kg, respectively) versus diabetes. The phytochemical analysis using GC-MS, HPLC, LC-MS
indomethacin group (41.50 ± 4.76), basal gastric secretion (5.00 ± 5.47 allowed the identification of numerous bioactive compounds, in
and 3.33 ± 5.16 at doses of 250 and 500 mg/kg, respectively) compared particular, terpenoids containing in essential oils of this plant. The
to control group (18.33 ± 1.50), and acidity (92.77 ± 12.36 and 64.10 variability between these components depends on plant part used and
± 7.20 mEq/L at doses of 250 and 500 mg/kg, respectively, versus the origin of plant. Few studies have determined the phenolic acids,
130.55 ± 8.27 mEq/L in control group). Moreover, treatment with flavonoids, and tannins containing in O. majorana extracts. However,
marjoram extract at the higher dose (500 mg/kg) significantly (p < investigations on these chemical families should be explored using
0.01-P<0.001) replenished the ethanol-induced depleted gastric wall different analytical techniques such as HPLC-DAD and LC-MS spectros­
mucus and non-protein sulfhydryls (NP-SH) (336.65 ± 18.97 and 3.55 copy tools. Furthermore, in vitro and in vivo pharmacological in­
± 0.29 μmol/g of Wet Glandular Tissue, respectively) compared to vestigations of O. majorana essential oils and extracts reported several
ethanol group 182.35 ± 12.46 and 2.40 ± 0.15 μmol/g of Wet Glandular effects such as antibacterial, antifungal, antiparasitic, antioxidant,
Tissue, respectively) and significantly (p < 0.01-P<0.001) lowered the antidiabetic, anticancer, anti-inflammatory, analgesic, anti-pyretic,
increase in the concentration of malondialdehyde (MDA) (1.96 ± 0.41 hepatoprotective, and antimutagenic activities.
μmol/g of Wet Glandular Tissue) compared to ethanol group (3.53 ± Essential oils of O. majorana (rich in carvacrol and thymol) showed
0.31 μmol/g of Wet Glandular Tissue). Histological examinations have remarkable bacteriostatic and bactericidal effects against several Gram-
confirmed the ulcer prevention of O. majorana ethanolic extract and Gram + bacteria. Further studies investigating the antibacterial
(Al-Howiriny et al., 2009). action of these EOs and their bioactive compounds against multi-
resistant bacteria are needed. The use of O. majorana EOs as additive
3.7. Toxicological investigation in food conservation could also reveal important results.
On the other hand, OMEO showed important antifungal effects, in
The acute toxicity study revealed that O. majorana ethanolic extract particular, against Candida albicans. However, the anti-candida action of
is quietly safe (Table 9) at different doses (1, 2, 3, 4, and 5 g/kg b.wt.) OMEO has not been mechanistically elucidated. In the light of this
(Seoudi et al., 2009). Indeed, no mortalities were recorded for result, it seems remarkable to thoroughly investigate the antifungal
O. majorana ethanolic extract in rats following oral administration in action of OMEO and their bioactive compounds against this pathogenic
doses up to 5 g/kg of body weight. In another study, Selim et al. (2013), strain and to elucidate the molecular mechanism both in vitro and in vivo
showed that essential oil of O. majorana did not present any toxicity on in experimentally infected animals. This could be used to screen for anti-
hamsters. Indeed, after 14 days of treatment, there was a total survival candida drugs from O. majorana.
and no symptoms were observed (Selim et al., 2013). The traditional uses of O. majorana were scientifically confirmed by
In order to complete the toxicity profile of this plant, further studies pharmacological assays, which linked the ethnopharmacological uses to
should be conducted to assess its toxicity over time and determine its the biological activities and secondary metabolite content, but without
effects on pregnant animals and their descendants, over long periods of investigating the actual mechanisms of action. The anti-oxidant effect of
study. O. majorana extracts and EOs is due, in particular, to the presence of
antioxidant bioactive compounds, essentially, phenolic substances.
Moreover, O. majorana showed remarkable pharmacological effects
against some oxidative stress related diseases such as cancer,

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A. Bouyahya et al. Journal of Ethnopharmacology 265 (2021) 113318

inflammation, and diabetes. In this way, the development of drugs from Al Dhaheri, Y., Eid, A., AbuQamar, S., Attoub, S., Khasawneh, M., Aiche, G.,
Hisaindee, S., Iratni, R., 2013. Mitotic arrest and apoptosis in breast cancer cells
O. majorana for diabetes, inflammation, and cancer needs further in vitro
induced by Origanum majorana extract: upregulation of TNF-α and downregulation
and in vivo tests to determine pharmacodynamic and pharmacokinetic of survivin and mutant. PloS One 8, 53.
parameters as well as the therapeutic efficacity. Al Tamimi, N.F., 2015. Anti-Colon Cancer Activity of Origanum Majorana.
Toxicological evidences revealed the safety of this plant on rate. Al-Fatimi, M., 2018. Volatile constituents, antimicrobial and antioxidant activities of the
aerial parts of Origanum majorana L. from Yemen. J. Pharmaceut. Res. Int. 1–10.
However, further investigations regarding the evaluation of toxicity at Al-Howiriny, T., Alsheikh, A., Alqasoumi, S., Al-Yahya, M., ElTahir, K., Rafatullah, S.,
different doses and different periods are required. Moreover, main 2009. Protective effect of Origanum majorana L. “Marjoram” on various models of
compounds of OMEO should also be tested for their toxicity. In this gastric mucosal injury in rats. Am. J. Chin. Med. 37, 531–545. https://doi.org/
10.1142/S0192415X0900703X.
regard, we invite research groups to carry out in-depth toxicological Alaoui, A., Laaribya, S., 2017. Etude ethnobotanique et floristique dans les communes
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validate its safety. On the other hand, in vivo evaluation of pharmaco­ Technol.
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dine El Omari, Douae Taha, and Taoufiq Ben Ali wrote a part of the Baâtour, O., Mahmoudi, H., Tarchoun, I., Nasri, N., Trabelsi, N., Kaddour, R., et al., 2013.
manuscript. Abdelaali Balahbib contributed to the collection of data and Salt effect on phenolics and antioxidant activities of Tunisian and Canadian sweet
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