Microbial Pathogenesis 134 (2019) 103580

Contents lists available at ScienceDirect

Microbial Pathogenesis
journal homepage: www.elsevier.com/locate/micpath

A comprehensive review of the antibacterial, antifungal and antiviral T

potential of essential oils and their chemical constituents against drug-
resistant microbial pathogens
Saika Tariqa, Saira Wania, Waseem Rasoola, Khushboo Shafia, Muzzaffar Ahmad Bhata,
Anil Prabhakarb, Aabid Hussain Shallaa, Manzoor A. Rathera,∗
a
Department of Chemistry, Islamic University of Science and Technology, Awanti Pora, Pulwama, Jammu and Kashmir, 192122, India
b
Instrumentation Division, Indian Institute of Integrative Medicine, Canal Road, Jammu, India

ARTICLE INFO ABSTRACT

Keywords: Essential oils are a complex mixture of odoriferous, volatile organic compounds. There are an extensive number
Antibacterial of published articles which highlight the antimicrobial action of a variety of essential oils from various parts of
Antifungal the world. The main aim of this review article is to compile these antimicrobial essential oils and their con-
Antiviral stituents from reliable sources and put them together. The published literature indicates that essential oils
Essential oils
possess a wide-spectrum of antibacterial, antifungal and even anti-viral activity. Essential oils have also been
Antibiotic
shown to inhibit the growth of drug-resistant microbial strains which are even difficult to be treated by con-
Terpenes
Infection ventional antibiotics. As for as their mode of action is concerned, in fungal pathogens, essential oils establish a
membrane potential across cell wall and disrupt ATP assembly, leading to cell wall damage. Essential oils can
also disintegrate mitochondrial membrane interfering with the electron transport system (ETS) pathway. In
bacterial pathogens, essential oils primarily destabilize the cellular architecture, leading to breakdown of
membrane integrity, disrupting many cellular activities including energy production and membrane transport.
Membrane rupture induced by essential oils can lead to leakage of cellular components and loss of ions. Several
essential oils have antiviral activities against many RNA and DNA viruses, such as type 1 and type 2 herpes
simplex virus (HSV-1 and HSV-2), dengue virus type 2, influenza virusadeno virus type 3, poliovirus, Junin virus,
and coxsackievirus B1. In conclusion, the current review article discusses in detail the various aspects of anti-
microbial activity of essential oils in a comprehensive manner.

1. Introduction perfumes, make up products as fragrances, household cleaning products

Essentail oils are volatile substances naturally produced by plants.
These are obtained from plant parts like flowers, buds, stems, leaves,
seeds, twigs, roots, fruits, bark and wood. Essential oils are stored in
cavities, secretory cells, epidermic cells, canals or glandular trichomes.
They are produced by plants as secondary metabolites, and are known
for their antibacterial, antifungal, insectcidal, antiviral properties.
Essential oils act against herbivores by triming their tendency for these
plants [1]. Essential oils constitute a major part of diverse natural flora
and are regarded as a major source in various fields like cosmetic,
fragrances, perfumery and pharmaceutical industries [2]. Currently
there are approximately 3000 established essential oils, some of them
(300) are important especially for food, sanitary, perfume, agronomic
industries e. g, g-limonene, gerenyl acetate or d-carvone are used in
and as industrial solvents [3]. These have also been used for oral and
dental treatments [4]. Owing to the fragrance and biological properties
of essential oils they have an overwhelming promising potential on
global market [5]. The special biological properties and fragrance of
essential oils are due to Terpenes and phenylpropanoids which are
major components of essential oils [6]. The presence of terpenes, phe-
nolics and aldehydes in EO's have a great application in bio-medicine as
they adequately snuff out many viral, fungal and bacterial pathogens
[7]. The antioxidative properties of EO's and their insect-repellent
properties have been confirmed [8]. Cardiovascular, neurodegenerative
disorders and cancer like diseases can be cured by certain bioactive
constitiuents of plants [9]. Sometimes the net biological activity of
essential oils cannot be ascribed to any of the major components and
the combination of components alter the biological activity to a greater

∗
Corresponding author.
E-mail address: manzooriiim@gmail.com (M.A. Rather).

https://doi.org/10.1016/j.micpath.2019.103580
Received 10 March 2019; Received in revised form 2 June 2019; Accepted 5 June 2019
Available online 11 June 2019
0882-4010/ © 2019 Published by Elsevier Ltd.
S. Tariq, et al.
extent for example, it has been reported that the inhibitory action of
rosemary oil against larvae insect is due to combined effect of various
EO constituents in which no individual constituent shows any biological
activity [10]. For centuries the use of essential oils in folk medicine and
the biological activity of a number of essential oils have been reported
by various studies. Essential oils have ability to accelerate the pene-
tration of drugs. The potential ability of certain essential oils is to en-
hance transdermal drug delivery. These are termed as sorption pro-
moters or accelerants and penetration enhancers. These essential oils
can penetrate the skin and are able to minimise the barrier resistance.
There are a number of mechanisms for the mode of action that have
been found for skin penetration enhancers; e. g, the interaction of liquid
crystals of skin lipids with essential oils [11]. Essential oils derived from
medicinal aromatic plants (e.g. Peppermint (Mentha piperita), thyme
(Thymus vulgaris), fennel (Foeniculum vulgare), are reported to be ef-
fective against Gram-negative and Gram-positive bacteria viruses, fungi
and yeast. EO's are reported to aid in defence mechanism in higher
plants [12]. Essential oils are the complex mixtures of natural com-
pounds of about 20–60 constituents in varying quantities. Major com-
ponents which are two or three in number nearly constitute 20–70% in
comparision to other components present in limited amounts because of
these major components the biological properties of essential oils are
generally determined. These components include two classes of sepa-
rate biological origin [13]. The prime group consists of terpenes and
terpenoids, the other of aliphatic and aromatic components and all have
low molecular weight.
As per monographs of International Pharmacopoeia the various
components of peppermint oil are menthofuran (1.0–9.0%), pulegone
(max. 4.0%), menthol (30.0–55.0%), carvone (max. 1.0%), cineole
(3.5–14.0%), pulegone (max. 4.0%), menthone (14.0–32.0%), limo-
nene (1.0–5.0%), isomenthone (1.5–10.0%), menthyl acetate
(2.8–10.0%), isomenthone (1.5–10.0%), limonene (1.0–5.0%), iso-
pulegol (max. 0.2%), menthyl acetate (2.8–10.0%), pulegone (max.
4.0%) and isopulegol (max. 0.2%) [14].
Rosemary essential oil was extracted by both steam and hydro-
distillation and the identification of components was done by com-
paring retention times to those of reference standards and their mass
spectra [15]. Essential oil from Lavender is extracted, normally by
steam distillation and there is a great diversity in chemical composition,
the most aromatic and sweeter oil being extracted from flowers [16].
The prime components of lavender essential oil are linalyl acetate, li-
nalool, β-ocimene (usually both trans and cis-), 1,8-cineole, terpinen-4-
ol, camphor and terpinen-4-ol [17].
2. Biological activity of essential oils
The essential oils inherent the activities of their major components
to show bio-activities [18]. Several studies have revealed the number of
the compounds phenols, aldehydes, ketones, alcohols, esters or hydro-
carbons present in essential oils manifests remarkable antimicrobial
property when tested separately and thus activity results from the
complex interactions between the different classes of compounds [19].
Essential oils containing aldehydes or phenols such as linanalde-
hyde, citral carvacrol, eugenol or thymol as major compound showed
highest antibacterial activity followed by essential oils containing ter-
pene alcohol. Other essential oil containing ketone or esters such as β-
myrcene, α-thujone or gerenyl acetate have much weaker activity.
While essential oils (volatile oils) containing terpene hydrocarbons are
usually inactive [20,21]. Essential oils bearing different terpenoid
components can interact in such a way that they either reduce or in-
crease their antimicrobial efficacy [22]. Because of interaction between
compounds they generate four possible types of effects: (a) indifferent,
(b) additive, (c) antagonistic or (d) synergistic effect [23]. When the
combined effects are equal to the sum of the individual effects this
observation is called additive effect. Antagonism is observed when the
effect of one or both compounds is less when they are applied together
2
Microbial Pathogenesis 134 (2019) 103580
than when individually applied. When the combined effect of sub-
stances is greater than sum of the individual effects the observation is
called synergism. While as absence of interaction is determined as in-
difference. The antimicrobial efficacy of essential oils depends on the
type of microbe to be inhibited as well as the evaluation methods, in-
cluding bioautography, difference and dilution [24]. Methods to eval-
uate the essential oil chemistry, their biological activities and various
factors that affect bioactivity are detailed in literature [25].
2.1. Antimicrobial effects of essential oils
Antimicrobial agents inhibit the growth of microorganism or lead to
their death. The antimicrobial effects of essential oils originated from
Medicinal and aromatic plants (MAP's) are basis of numerious appli-
cations, in various revenue generating sectors such as pharmaceuticals,
nutraceutical, cosmetic, perfume, agronomy and sanitary industries.
Most recently the prevalence of antimicrobial drug resistance has be-
come the new challenge for researchers to develop noval microbial lead
molecule to target various human pathogens [26]. As the inhibition of
many pathogenic microbes by using synthetic drugs fails and further-
more the uses of synthetic chemicals for control of pathogenic micro-
organism is concised due to the reason of carcinogenic effects, acute
toxicity and environmental hazard potential of these synthetic drug.
Essential oils are gaining more popularity because many synthetic drug
are connected with unpleasant side effects, such as nephrotoxicity or
ototoxicity. Volatile oils represents an interesting alternative due to
emerging resistance of microorganism against synthetic drugs. That is
reason why?
Essential oils are succeeding in inhibiting the microbial growth of
drug resistant bacterial strains. The antimicrobial activity of essential
oils can be witnessed by invitro tests, several methods are used to in-
vestigate the antimicrobial activity of essential oils. And most three
important ones are the agar diffusion, the agar or broth dilution and the
vapour phase test [27]. In this view exploitation of essential oils to
control multi drug resistant pathogenic microorganism can be suffi-
ciently used to combat various infectious diseases. In the following
section we have broadly elucidated the antibacterial, antifungal, anti-
viral potentials of essential oils extracted from MAP's as well as their
therapeutic revelance and possible mechanism involved in eradication
of pathogenicity in humans.
2.2. Antibacterial effects of essential oils
The various bacterial pathogens are treated by using the common
antibiotics. But nowadays the bacterial pathogens become resistant to
these multidrug antibiotics which led to the increase to severity of
diseases and therefore the scientists are now facing a challenge to find
the alternate way for the treatment of such diseases. Furthermore the
bacteria have ability to form bioflim associated drug tolerance and also
weak immunity in host cell leads to increase in number of life threa-
tening bacterial infection in human body [28]. Hence, today the major
causative agents for the number of human deaths are due to the bac-
terial infections in addition, higher doses of several antibacterial drugs
causes toxicity in the human beings. In the recent years, there has been
a growing interest in researching and exploring new and alternate key
molecules from various sources to combat the drug tolerance bacterial
strains [29]. In the view, essential oils obtained from the plants and
their major chemical composition are potential candidates to fight
against the bacterial infections [antibacterial agents]. Table 1 shows the
several type of essential oils and their major chemical constituents
obtained from various MAP's that have been reported and posses a wide
range of bacterial inhibitory potential.
The antibacterial activity of essential oil have severe effect as they
may seize the growth of the bacteria (bacteriostatic) or kill bacterial
cells (bactericidal). The minimum concentration of antimicrobial agents
that completely inhibits growths of the organism in micro-dilution wells
S. Tariq, et al. Microbial Pathogenesis 134 (2019) 103580

Table 1
Chemical composition of various essential oils and their antibacterial activity human pathogen.
MAPs Part used Major chemical compound Inhibited microorganisms Reference

Artimisia cana Aerial parts Santolina triene, α pinene, camphene Escherichiacoli, Staphylococcusaureus, Staphylococcus epidermidis [30]
Achillea ligustica Aerial parts Viridiflorol, terpin-4-ol Streptococcus mutans [31]
Artimisia frigida Aerial parts 1,8-cineole,methylchavicol, camphor E.coli, S. Aureus, S.epidermidis [30]
Achillea clavennae Leaves and Camphor, myrcene,1,8-cineole, β- Klebsiella pneumonia, Streptococcus pneumonia, Haemophilus influenza, [32]
Flowers caryophyllene,linalool, Gerenyl acetate Pseudomonas aeroginosa
Cyperus longus Aerial parts β-Himachalene,α-humulene, S.aureus, Listeria monocytogenes,, Enterococcus faecium, Salmonella [33]
γ-himachalene enterica,E.coli, Pseudomonas aeruginosa
Cuminum cyminum Leaves γ-Terpin-7-al,γ-terpinene,β-pinene, Salmonella typhimurium,E.coli [34]
Cuminaldehyde
Copaifera officinalis Essential oil δ-cadinene,germacrene D, E.coli, S.aureus [34]
α-humulene,α-copaene, germacrene B,β-
caryophyllene,
β-bisabolene
Cymbopogon citrus Leaves Ethanolic compounds S.aureus, Enterobacteriaceae [36]
Cymbopogon citrus Leaves Ethanolic compounds S.aureus, Enterobacteriaceae [36]
Cymbopogon citrus Leaves Ethanolic compounds S.aureus, Enterobacteriaceae [36]
Dracocephalum foetidum Leaves limonene, n-mentha1, Enterococcus hirae,S.aureus, Micrococcus luteus,E.coli, [37]
8-dien-10-al Bacillus subtilis, Streptococcus mutans
Eugenia caryophllata Flower buds Thymol, eugenol, carvacrol, cinnamaldehyde S.epidermidis [38]
Eremanthus erythropapps Leaves viridiflorol, p-cymene germacrene D, γ- S.epidermidis [39]
terpinene (Z)-caryophyllene
Foeniculum vulgare Leaves limonene, methylchavicol, E.coli, Salmonella typhimurium [34]
Trans-anthole
Juniperus phoenicea Arial part α-terpinyl actate,α- pinene P.aeruginosa,E.coli, S.aureus,E.faecium, Salmonell Enteriditis [33]
β-phellandrene
Mentha piperita Aerial parts _ S.typhimurium,S.aureus, Vibrio parahaemolyticus [40]
Momordica Charantia Seed germacrene D, Trans nerolidol, S.aureus,E.coli [41]
cis-dihydrocarvacol
Laurus nobilis Arial part linalool, Eucalyptol (1,8-cineole) Ecoli. Mycobacterium smegmatis [42]
Lippia sidoides Leaves carvacrol and thymol Streptococcus salivarius, Streptococcus sanguis Streptococcus mitis [43]
Streptococcus mutans
Nigella sativa Seeds longifolene, Thymoquinone P.aeruginosa,S.aureus E.coli, Bacillus cereus [44]
thymohydroquinone,
α- thujene,p-cymene
Ocimum basilicum Leaves, stems methylchavicol, γ-terpinene Pseudomonas putida, Mariniluteicoccus flavus, Listeria innocua,E.coli [45]
S.typhimurium, Brochothrix thermosphacta
Pimpinella anisum Seed Trans-anethole E.coli,S.typhimurium [46]
P.amboinicus Leaves viridiflorol, γ-terpinene, germacrene D,p- S.epidermidis [47]
cymene (Z)-caryophyllene
Rosmarinus officinalis Leaves, flower linalool, borneol, limonene, camphene, Bacillus subtilis,S.aureus, E.coli,S.epidermidis, B.cereus,P.aeroginosa
myrcene, camphor, gereniol, Listeria innocua, Enterococcus faecalis, pseudomonas putidaMycobacterium [48,49]
α-pinen, bornyl acetate, smegmatis,
α-terpinolene, linalool benzoylacetate S. typhimurium, Strongylus vulgaris
Salvia officinalis Arial part 1,8-cineole,camphor, E.coli, Providencia stuartii
α-pinene,α-thujone Sarcina lutea,S.aureus, Shigella sonnei,M.flavus, L.innocua, [50,51]
Syzygium aromaticum Leaves eugenylacetate, Eugenol Enterobacteriaceae, P.aeruginosa [35]
Salvia lavandulifolia Essential oil camphene, terpineol, Enterococcus faecalis,P.aeruginosa P.vulgaris, Klebsiella pneumoniae [52]
α-pinene,α-thujone camphor, β-thujone
Trachyspermum ammi Seeds _ S.aureus,E.coli, K.pneumoniae [53]
Thymus zygis Seeds _ E.coil,S, typhimiriu, Salmonella choleraesuis [54]
Warionia saharae Aerial part terpine-4-ol, p-cymene, P.aeruginosa,B.cereus [55]
trans-nerolidol,camphor, E.coli, S.aureus
1,8-cineole,linalool,
β-Eudesmol

or tubes as detected by unaided eye is called Minimum Inhibitory
Concentration (MIC) [56]. Thus the estimation of bactericidal activity is
determined as minimum bactericidal concentration [MBC], which is
defined as concentration killing 99.9% or more of initial inoculums
[57]. The rapid screening action of essential oils on bacterial agents is
usually conducted using agar diffusion technique. The effectiveness of
essential oils differ from one type to another depending on bacterial
infections as well as against different target bacteria (Gram positive and
Gram negative) [58]. Former studies showed a higher antibacterial
effects of essential oils against Gram positive rather than Gram negative
[59]. For example vetiver oil and sandal wood oil fails to inhibit Gram
negative bacterial strain but shows higher inhibitory action against the
Gram positive bacterial strains. Essential oils and their components are
gifted by important characteristic i. e hydrophobicity. Which enables
essential oils to separate them with lipids present in cell membrane of
bacteria and mitochondria rendering them more permeable by dis-
turbing the cell structure. This ultimately leads to the death of bacterial
cell due to leakage of critical molecules and ions from bacterial cell to
great extent. Some compounds modulate drug tolerance by attacking
efflux mechanism in several species of Gram negative bacteria [60]. The
outer cell membrane of Gram negative bacteria possesses hydrophilic
properties that obstruct the contact of hydrophobic constituents of es-
sential oils with bacterial cell [61]. In contrast to this essential oils
directly damage cell membrane of Gram positive bacteria resulting in
rupture of cell membrane by blockade of enzymes system and pro-
gressivity of ion permeability. The essential oil obtained from clove,
cinnamon, oregano, pimento, rosemary and thyme have shown strong
antibacterial activity against Staphylococcus aureus, Salmonella typhi and
Pseudomonas aeruginosa [62]. The most effective among all tested es-
sential oil was founded clove oil. The antimicrobial effect of these

3
S. Tariq, et al.
essential oils was proportional to the availability of major compounds
such as eugenol, cinnamic aldehyde, thymol, p-cymene and carvacrol
[63]. The compounds like benzaldehyde, benzoic acid and cinnamic
acid condition under anaerobic showed up to 50% inhibition of Listeria
monocytogenes. Clove oil has inhibitory effect on Listeria monocytogenesis
as it effects on respiratory metabolism, structural changes of DNA with
interaction of eugenol (main component of clove oil) and cell mem-
brane permeability [64]. The essential oils from basil, cardamom, ro-
semary, anise, coriander, angelica, drill and parsley and their efficacy
against saprophytic and pathogenic microorganism was scrutinized by
Elgayyer et al. [65]. The ample use of essential oils or their utilization
at high concentration Hood et al. [66] delineated the arrestment of
bacterial growth and the mode of action of essential oils results in the
decrease in bacterial cells. The MIC value of 0.25% to ≥2% v/v is
shown by an essential oils oregano and thyme against pathogenic
bacteria; Salmonella enteritidis, E. coli, Salmonella choleraesuis and Sal-
monella typhimurium [67]. Camphor,α-Thujone, and 1,8- cineole are the
major chemical components of essential oil of Salvia officinalis and has
the inhibitory activity in human bacterial pathogens such as Providencia
stuartii and Salmonella aureus. [68]. As per Nevas et al. thyme, oregano
and savory essential oils effectively inhibited the pathogenic bacteria
such as Clostridium perfringens and Clostridium botulinum. Achillea ligus-
tica a medical plant; contains 1,8- cineole, terpinen-4- ol, linalool and β-
pinene showed effective inhibitory activity against S. mutans with an
MIC range of 155–625 μg/mL. Ouazzou et al. Studied Mentha pulegium
an essential oil has the best antibacterial activity compared to Juniperus
pheonicea and cyperus longus1μg/mL MIC value for E. coli, S. enteritidis,
S. aureus and L. monocytogenes. M. pulegium oil has the MIC value < 0.5
for Enterococcus faecium. The essential oil of cinnamomum zeylancium
inhibits the diverse range of bacterial pathogens such as Enterococcus
feacium, streptococcus pyogenes, S. aureus, S. pneumoniae, B. cereus, En-
terococcus faecalis, Acinetobacter lwoffii, K. pneumonia, E. coli, E. aero-
genes, S. typhimurium, Proteus mirabilis, Mycobacterium smegmatis, P.
aeruginosa and C. perfringens reported by Unlu et al. [69]. The patho-
genic bacterial strains such as Neisseria gonorrhoeae, E. coli, B. subtilis, S.
aureus and P. aeruginosa the activities of all these were effectively in-
hibited by the essential oils of Syzygium cumini which contains trans-
caryophylene, limonene, 1,3,6- octatriene, β-pinene, δ-3- careen, α-
pinene, and α-caryophyllene as major compounds and responsible for
effective antibacterial activity [70]. In aromatheraphy procedures An-
drade et al. studied 27 different essential oils with antimicrobial ac-
tivity contain Cinnamomum cassia, Melaleuca alternifolia, Piper nigrum
and Copaifera officinalis essential oils against E. coli and S. aureus
whereas essential oil S. aromaticum was efficient against P. aeruginosa
strain. (Tulsi) Ocimum tenuiforum of Australian grown essential oil re-
vealed antimicrobial activity against microbial pathogens like (MRSA)
Methicilin resistant S. aureus, P. aeruginosa and E. coli with the in-
hibitory zone ranging from 2.25 to > 4.5 μg/mL [71]. Herb Struchium
sparganophora produces essential oil which shows the existence of
germacrene A, - caryophyllene, germacrene D and α-humelene as a
major chemical constituent and inhibits the P. aeruginosa, B. subtilis, S.
typhi, P. mirabilis and B. cereus [72]. The MIC ranging from
18.5 ± 2.2 mm to 20.0 ± 0.0 mm obtained essential oil from stem
while as MIC value ranging from 9.0 ± 1.0 mm to 14.3 ± 2.55 mm
when obtained from leaf oil. A good antimicrobial activity is seen in
onion which inhibits S. aureus (MIC = 12 μg/mL). The best activity is
shown by Chamomile (Anthemis nobilis) oil againt P. aeroginos
(MIC = 5.1 μg/mL) Mahmoud et al. [73]. All 36 clinical isolates of E.
coli, S. aureus and K. pneumonia isolates from patients of suffering from
urinary tract infections were inhibited by the essential oil of Trachy-
spermum ammi which is extracted from seeds [74].
2.2.1. Mechanism of antimicrobial action of essential oils against human
pathogens
MAP's contain several types of chemicals constituents that have
antimicrobial properties. These are synthesized to protect the plants
4
Microbial Pathogenesis 134 (2019) 103580
from microbial pathogens. The antimicrobial property of essential oils
mainly depends on their chemical constituents and the quantity of
major single compounds. These chemical compounds are secreted
through a series of molecular interactions under specific biotic/abiotic
stress conditions [75,76]. Each compound may exhibit a different me-
chanism of antibacterial action is mediated by a series of biochemical
reactions in the bacterial cell, which are dependent on the type of
chemical constituents present in the essential oil [77]. Moreover, the
antibacterial activity of essential oils also differs because of different
bacterial architecture, such as Gram-positive and Gram-negative bac-
terial, which differs in their cell membrane compositions [78,79]. In the
following sections, the mechanism of antimicrobial activities of essen-
tial oils is described with reference to the available literature.
2.2.2. Action against selected bacterial pathogens
Various mechanisms of antibacterial activity of essential oils have
been proposed. Essential oils primarily destabilize the cellular archi-
tecture, leading to the breakdown of membrane integrity and increased
permeability, which disrupts many cellular activities, including energy
production (membrane-coupled), membrane transport, and other me-
tabolic regulatory functions. The disruption of the cell membrane by
essential oils may affect various vital processes, nutrient processing, the
synthesis of structural macromolecules, and the secretion of growth
regulators [80]. The essential oils may affect both the external envelope
of the cell and cytoplasm. Owing to their lipophilic nature, essential oils
are easily penetrable through the bacterial cell membranes. Essential oil
of various MAPs were reported to cause increased bacterial cell,
membrane permeability leading to the leakage of cellular components
and loss of ions [80,81]. The antibacterial effect of essential oils is also
linked to reduce membrane potential, the disruption of proton pumps,
and the depletion of ATP [82]. This alteration in the cell organization
may cause a cascade effect, resulting in the cell organelles being af-
fected [83]. Likewise, Cox et al. [84] have demonstrated that tea tree
oil inhibits the growth of S. Aurous and E. Coli by altering cell per-
meability, increasing the leakage of intracellular K+ ions and disturbing
cell respiration. The essential oils pass through cell wall and cytoplasm
membrane, which may disrupt the arrangement of dissimilar fatty
acids, phospholipids bilayers, and polysaccharides molecules [85]. All
these events may be responsible for the coagulation of inner cellular
components in the cytoplasm and breakdown of the bonds between the
lipid and protein layer. In some cases, the pure compounds of essential
oils exhibits higher antibacterial activity compared to the essential oil.
The antibacterial effect of essential oils constituents such as thymol,
menthol, and linalyl acetate is because of perturbation of the lipid
fraction of bacterial plasma membranes [86]. This may affect the per-
meability of the membrane and induce leakage of intracellular mate-
rials. Carvacrol is a hydrophobic compound that influences cell mem-
brane by altering the composition of fatty acids which affects the
membrane fluidity and permeability [87]. However, its extra me-
chanism of action is still unclear. It was reported that carvacrol sig-
nificantly depleted the internal ATP pool of the bacterial cells [88]. In
another study carvacrol induced the leakage and loss of ATP from
bacterial cells [89]. Likewise, the compounds methyl carvacrol,
methnol, citronellol, and thymol also cause an enlargement of the cell
membrane that leads to passive diffusion of ions between the expanded
phospholipids. Another effect of essential oils on cell membranes is the
inhabitation of toxin secretion. Ultee and Smid reported that exposure
of B. Cereus to carvcrol resulted in the inhibition of toxin production,
and application of oregano essential oil completely abolished the en-
terotoxin production of S. Aureus. Thus, the secretion of toxin may be
prevented by modifications in the bacterial membrane transport pro-
cess in the plasma membrane due to the influence of the essential oil
compounds on the trans-membrane transport process in the plasma
membrane, which limits the release of toxins to the external environ-
ment [90]. Another mechanism of action by trans-cinnamaldehyde,
which enters the periplasm of the cell and disrupts cellular functions
S. Tariq, et al.
[91]. Moreover, p-cymene has a greater affinity towards bacterial cell
membranes and thus may be disturb the membrane integrity [92]. The
outer membrane proteins are also affected by essential oil components.
For example: carvacrol can disturb the insertion and folding of proteins
such as DnaK and GroEL. Carvcrol can also inhibit the synthesis of
flagellin, a microbial protein required for bacterial motility. The phe-
nylpropene, eugenol, also exhibits activity by modifying the fatty acid
outline to alter the cytoplasmic membrane of different bacteria. In
addition it can destroy various bacterial enzymes such as ATPase,
amylase histidinecarboxylase, and proteases [93,94]. Likewise, cinna-
maldehyde was reported to inhibit ATPase enzymes and disrupt the
outer cell membrane. Other studies have found that vanillin exhibited
antimicrobial activity by obstructing the pathways of bacterial re-
spiration and disrupting the flux K+ ions and pH gradient. Similarly
careveol, citronellal, and carvone essential oils were shown to modify
hydrophobicity and disrupt membrane integrity, leading to the leakage
of K+ ions [95]. Some essential oils can inhibit the cell-cell commu-
nication quorum sensing network ediated by various bacterial signal
molecules [96]. The efficacy of the antibacterial effect of essential oils
or their individual compounds may differ from one microbe to another.
Hence elucidation on the exact mechanisms of action of each essential
oil and their components is required, including further study on the
numerous microbial strains/species. Furthermore, detailed study of the
components of the essential oils would be helpful to improve our un-
derstanding of their mechanism of antimicrobial activity. Fig. 1 depicts
the antimicrobial mechanism of essential oils on microbes and different
uses of essential oils are represented in Fig. 2.
2.2.3. Essential oils against drug-resistant bacterial strains
A new challenge for scientists has emerged to find possible mode for
cure of infections, which are caused by the tolerance of several mi-
croorganism against commonly used antibiotic drugs. The imprecise
application of drugs is one of the major reasons that provokes the
higher resistance to microorganisms [97]. (E. coli), Enterobacter aero-
genes Pseudomonas aeruginosa and Acinetobacter baumannii are some of
the multidrug resistant Gram negative bacterial strains whose action is
inhibited by gereniol, one of the constituent of Helichrysum italicum
(Roth) G. Don fil.(Asteraceae) volatile oil. It was assumed that this es-
sential oil contains substances that acts as efflux pump inhibitors.
Fig. 1. Antimicrobial mechanism of essential oils on microbes.
5
Microbial Pathogenesis 134 (2019) 103580
Essential oils revealed to be active against bacteria which over-ex-
pressed efflux pumps and therefore developed tolerance towards drugs
[98]. A common part of human microbial skin flora– S. aureus; a Gram
positive bacterium– causes minor infections; nevertheless, also causes
severe diseases like meningitis, sepsis, pneumonia and endocarditis espe-
cially in hospitalized patients. These pathogens have increased re-
sistance to the current drugs, complicates tremendously the therapy of
these infections [99]. The antimicrobial active agent terpinen- 4-ol, the
constituent of Melaleuca alternifoliaCheel.(Myrtaceae) ethereal oil. The
bactericidal and bacteriostatic activity of both terpinen-4- ol isolated
from M. altrernifolia and volatile oil M. alternifolia in vitro tests has been
ascertained against Methicillin-resistant staphylococcus aureus(MRSA)
and coagulase-negative staphylococci shows much stronger activity
when using terpinen-4-ol on its own. As a result an interesting alter-
native could be terpinen-4-ol that can be used in the therapy of skin
infections MRSA [100]. Hence as a results of above, it supports the idea
of using volatile oils to form well established drugs which show high
efficacy to inhibit drug-resistant bacterium strains. Thus essential oils in
combination with synthetic active agents as synergy was seen or in
combination with other essential oils as well as ethereal oils on their own
could be used. Essential oil Zataria multiflora in combination with the
synthetic active agent vancomycin [101] as well as when combining
Lavandula luisieri essential oil with L. angustifolia or L. stoechas essential
oil were observed to have synergistic effect against MRSA. One of the
natural essential oils litsea cubeba has destructive effect on cell mem-
brane and Hexose monophosphate pathway (HMP) of MRSA [102].
Table 2 shows essential oils and methicillin-resistant Staphylococcus
aureus (MSRA). The Myrtaceae family whose members are Melaleuca
alternifolia, Eukalyptus globulus, C. operculatus, the essential oils of
whose monocyclic monoterpenes like (terpinen-4-ol and 1,8-cineole)
shows effectively inhibition efficacy against the growth of drug-re-
sistant bacterial strains.
2.2.4. Essential oils against helicobacter pylori (H.pylori)
In the stomach of many humans, the Gram-negative bacteria
Helicobacter pylori colonizes. The symptoms on one hand may be gas-
tritis and ulcers but on other hand this infection can proceed without
symptoms. Proton-pump inhibitors in combination with antibiotics are
commonly treated for these complications [107]. Although essential
S. Tariq, et al.
Fig. 2. Flowchart representation of the use of essential oils in different industries.
oils like Thymus caramanicus, Apium nodiflorum, Plinia cerrocampanensis,
Dittrichia viscose could inhibit the growth of this pathogens as tests, in
the last three years relatively few studies have dealt with the effect of
essential oils against H. pylori. Being the fact that the individual volatile
oils does not share characteristics considering their family origin as well
as their chemical composition, they possess the strong antimicrobial
activity against this pathogen in common. In general the secure and
effective eradication of this pathogen is enabled by the combination of
several synthetic drug. The answer to this question why few studies are
conducted to find essential oils as agents against H. pylori. Table 3.
Shows essential oils possess antibacterial activity against H. pylori.
2.3. Antifungal effects of essential oils
The essential oils and their constituents are used against a broad
range of plant pathogens. The essential oils extract from several plants
like basil, citrus, fennel, lemon grass, oregano, rosemary, and thyme have
shown significant antifungal activity against a large varietyof flora
pathogens [111]. The antimicrobial activity of essential oils extracted
from spices against fungal pathogens observed by Arora and Kaur
[112]. They found that garlic and clove extracts inhibited the expansion
of Candida acutus, Candida albicans, Candida apicola, Candida catenulata.
Candida inconspicua, Candida tropicalis, Rhodotorula rubra, Saccharomy
cerevisiae and Trigonopsis variabilis. According to the report of Ultee and
Smid [113], oregano and thyme essential oils were some of the best
inhibitors of fungal pathogens, because of the phenolic compounds
(carvacrol and thymol) as main constituents, which disrupt fungal cell
membranes. Likewise, Delaquis and Mazza [114] reported the anti-
microbial effects of chemical irritant isolated from the essential oils of
onion and garlic plants. They expressed that isothiocyanates may
Table 2
Antibacterial activity of Essential oils/their components against Methicillin- Resistant Staphylococcus Aureus(MRSA).
Effective against Essential oil
MRSA,VRE Kadsura longipedunculata Fine & Gagnepain(Schisandraceae)
MRSA Salvia rosifolia Sm.(Lamiaceae)
MRSA Lavandula latifolia Medik (Laminiaceae)
MRSA Thymus vulgaris L.(Laminaceae)
MRSA Eucalyptus globules Labill.(Myrtaceae)
MRSA Lavandula luisieri Rozeira Riv.-Mart. (Laminaceae)
MRSA Zanthoxylum tingoassuiba St.-Hil. (Rutaceae)
Microbial Pathogenesis 134 (2019) 103580
inactivate the extracellular enzymes through oxidative cleavage of
disulphide bonds. Isothiocyanate was effective against Botrytis, Fu-
sarium, fungus genus, and Cladosporium species. The antifungal activity of
essential oils and their derivatives on the cell viability, plant growth,
and mycotoxin-producing ability of moulds has been studied [115].
More recently, Eugenol (a volatile oil compound from clove) was shown
to cause permanent injury to the cells of C. albicans and was thought to
be an economical antifungal (Table 4).
2.3.1. Mechanism of action of essential oils against fungal pathogens
In yeast cells, essential oils establish a membrane potential across
the cell wall and disrupt the assembly of ATP, that ends up in cell wall
damage [116]. The essential oils have the power to penetrate and dis-
rupt the fungal cell wall and protoplasm membranes through a per-
meabilization process, that ends up in the disintegration of mitochon-
drial membranes. This can be caused by adjustment within the flow of
electrons inside the electron transport system (ETS) pathway. This
might conjointly harm the lipids, proteins and supermolecule contents
of cells infected by the fungal pathogens [117]. The action of essential
oils against Gram-positive bacteria and fungi appears to be similar in
sort. The oil elements destroy the microorganism and fungal cell wall
and living substance membrane, which ends up in an exceedingly run of
the protoplasm and its action [118]. In some fungi and alternative
gram-positive microorganism, which are sensitive to iminazole and
whose cell membranes are rich in unsaturated fatty acids, the mem-
brane constituents arrangement leads to the loss of cell viability and,
eventually, lysis. Essential oils additionally inhibit the synthesis of
DNA, RNA, proteins and polysaccharides in flora and micro-organism
cells [119,120]. In fungi, and become ineffective as a result of the
speedy development of fungal resistance [121]. The effects that occur
Main component Test method Ref.
δ-Cadinene (21.8%) Diffusion test, dilution test [103]
α-Pinene, 1,8- cineole MIC = 125 μg/Ml [104]
Linalool (38.8%); 1,8-cineole Disc diffusion [102]
Thymol (48.1%) MIC = 18.5 μg/mL [105]
1,8- cineole (47.2%) MIC = 85.6 μg/mL [104]
α-Necrodyl acetate(34.5%); 1,8-cineole (17.6%) Disc diffusion [102]
α-Bisabolol, methyl-N- methylanthranilate Disc diffusion [106]
6
S. Tariq, et al. Microbial Pathogenesis 134 (2019) 103580

Table 3
Essential oils against Helicobacter pylori
Effective against Essential oil Main component Test method Ref.

Thymus caramanicus Carvacrol (68.9%) Disc diffusion testMIC = 14.5–58.0 μg/mL [108]
H.pylori
Apium nodiflorum(L.) Lag.(Apaeceae) Limonene(27%); p-cymene (23%); Myristicine (18%) MIC = 12.5 μg/mL [109]
H.pylori
Plinia cerrocampanesis Barrie (Myrtaceae) α-Bisabolo (42.8%) MIC = 62.5 μg/mL [110]
H.pylori

due to essential oils are similar to the effects of antibiotic action [122].
Natural antifungal agents are in great need because currently used
therapeutics have toxic side-effects which, may interact with other
drugs.
Trichophyton may be a plant life species that causes superificial
mycoses unremarkably referred to as tinea infections in varied areas in
humans and alternative animals. Ketoconazole is amongst the un-
remarkably used antifungal medication administered orally for the
treatment of each superificial and deep infections caused by
Trichophyton. However, the unpleasant side-effects of this drug embrace
nausea, abdominal pain and skin sensation, and its toxicity limits its
therapantic use in several cases. Futhermore, the therapeutic response
may be slow, and thus unsuitable for treatment of patients with severe
or rapidly progressive mycoses. Additionally, the efficacy of ketoco-
nazole is poor in immunological disorder patients and within the
treatment of infectious diseases. EO's are one amongst the foremost
promising teams of natural compounds for the event of safer antifungal
agents. However their poor absorption from the human bowel and
comparatively delicate antifungal activity compared with industrial,
artificial antifungal medication could ultimately limit their clinical
application within the treatment of general plant life infections [123].
Many essential oils are helpful for treatment of tinea and their in
vitro and in vivo antifungal effects are evaluated [124]. In general, their
antifungal activity is delicately compared with normally used anti-
biotics. Several essential oils are only fungistatic and high concentra-
tions are required for agent activity [125]. To reinforce the efficacy of
EO's, the combined use of various oils, has been evaluated recently for
potential synergistic effects. The mixture of EO's with artificial anti-
fungals will possibly result in a more effective therapy though. However
synergistic effect between EO's and customary antifungal medicine
against tricho-phyton haven't yet been reported until now so far. So as to
develop a broad spectrum natural anti-genus trichophyton agent [126].
We tend to evaluate the activities of EO's used for the treatment of plant
life infections in aroma-medical core and practice of medicine against
six species of genus trichophyton fungi by broth dilution & disc diffusion
tests. Additionally we have a tendency to evaluate the synergistic ef-
fects of the EO's from Pelargonium graneolens, a potent oil containing the
extremely active & stable primary alcohols citronellol & Geranoid &
ketoconazole, a normally used therapeuntic for Trichophyton infections
against Trichophyton spp.
2.3.2. Growth inhibition of Trichophyton spp. On sabouraud's agar plates
The consequences of the disc diffusion tests (Table 5) also indicate
that these oils are significantly potent against Trichophyton spp. Con-
sistent with the MIC assay results, the oils of Cymbopogon citrates, La-
vandula angustifolia, Pelargonium graveolens, Rosmarinus officinalis and
Thymus vulagris exhibited rash examine of > 39 mm in most out-
standing of the extremist concentrations of 25 and 12.5 mg per disc.
The results from tests with T. vulagris and its main part thyme camphor
and P. graveolens and its main components citronellol and geraniol were
similar, at 1.25 mg per disc. However, not all the fungal inhibition re-
sults on sabourand's agar plates were consistent results from the MIC
assays. For example, the radius of inhibition by Eukalyptus globulus oil
and its main component 1,8-cineol was smaller than expected based on
its M.I.C.Funiperus communis and Pogestemon patchouli oils were rela-
tively ineffective against all the fungi tested, with inhibition zones be-
tween 2.8 and 7.3 mm even at 25 mg per disc sabouraud's dextroglucose
agar plates. The widths of the fungous inhibition zones were usually
dose dependent.
The increasing resistance to antifungal compounds and the reduced
number of available drugs led us to search for the new alternatives
among aromatic plants and their essential oils, used for their antifungal
properties.
The antifungal activity may be attributed to the presence of some
elements like carvacrol,α-terpinly acetate, cymene,thymol, pinene volatile
oil that area unit already well-known to exhibit antimicrobial activity
[127]. Variety of scientific investigations have highlighted the im-
portance and therefore the contribution of the many plant families
i.e.Asteraceae, Liliaceae, Apocynaceae, Solanaceae, Rutaceae, Piperaceae
etc, used as healthful plants [128]. Numerous in Vitro studies have been
posted confirming the impact of crucial oil and their predominant
compounds on plant and human pathogenic fungi. Some of the plant
families and their antifungal activity of volatile oil are summarized
below in Table 6.
Fungal infections are caused by eukaryotic organisms, and it's so
harder to establish their presence and apply the suitable therapeutic

Table 4
Minimum Inhibitory concentration (MIC) and minimum fungicidal conc. (MFC) of herbal oils against Trichophyton spp.
FUNGI.
E.O Trichophytonerinacei T.Mentagrophytes T.rubrum T.schoenleinii T.soudanense T.tousurans

MIC MFC MIC MFC MIC MFC MIC MFC MIC MFC MIC MFC

Citrus Bergamia 4 16 2 4 1 4 2 4 0.5 2 1 2

Cedrus Atlantica 2 4 1 2 1 2 1 2 0.25 0.5 0.5 1
Geraniol 0.5 2 0.5 1 1 1 0.5 1 0.25 0.5 0.5 1
Citronellol 0.5 4 1 2 2 4 1 4 0.5 1 2 4
Thymus vulgaris 0.5 1 1 1 1 1 0.5 1 0.5 1 1 2
Benzoic acid < 0.125 0.125 0.25 0.5 0.25 < 0.25 < 0.25 < 0.25 0.25 0.5 0.25 0.5
Thymol 0.25 0.5 0.5 1 0.5 1 0.5 1 0.5 1 0.5 1
Eukalyptus globulus 0.25 1 0.25 0.5 < 0.125 0.5 0.25 0.5 0.25 0.5 < 0.125 0.125
Lavandula Angustifolia 0.5 4 2 4 0.5 4 2 4 0.25 0.5 1 2
Melaleuca Alternifolia 0.5 3, 12 1 > 32 1 > 32 2 > 32 8 16 1 2

7
S. Tariq, et al.
treatment compared to micro-organisms infections. The chitin structure
of fungi present in cell wall is regarded as the prime target for selec-
tively toxic antifungal agents which is absent in humans. Chemical
treatments are mostly effective, however resistant strains and in-
trinsically resistant species are often developed. The onset and severity
of the fungal infection depends on the substance charge, the host's
medical specialty state and resistance. Essential oils can represent one
of the most promising natural products for flora inhibition [152,153].
The antimicrobial or antifungal activity of EO's could be caused by the
properties of terpenes/terpenoids, that due to their extremely oleophilic
nature and low molecular weight –are capable of disrupting the cell
wall, causing cell death or inhibiting the separation and propagation of
food spoilage fungi. Therefore many in-vitro tests indicates that ter-
penes/terpenoids show ineffective antimicrobial activity once used as
singular composite as compared to the complete EO [154,155]. All the
activities exhibited by E. O's &/or their part, which can be mentioned in
this Table 7.
According to Freiesleben and Jager [216], the antifungal agents will
deactivate the plant life by disrupting the structure and performance of
membranes or organelles of plant cell and/or inhibiting the nuclear
material or macromolecule synthesis (Fig. 1).
During the previous couple of decades, plant infections are thought
to be a heavy unhealthyness and a serious sickness, significantly among
disorder patients. Talking regarding immunocompetent people, mycosis
(tinea or ringworm) could be a prevailing variety of zymosis often en-
countered in these people in medical speciality patient department.
This infection is caused by a bunch of keratinophilic fungi referred to as
dermatophytes that affects the keratinous tissues of humans and dif-
ferent vertebrates, inflicting superficial infections. Dermatophytes be-
long to 3 genera: Trichophyton, Microsporum and Epidermophyton [217].
Mycosis will transfer from soil and animals to humans and cause in-
fection on several elements of the body [218]. The antifungal medicine
most typically used against these diseases embody antibiotic ketoco-
nazole, fluconazole, terbinafine and flucytosine. Adverse reactions are
related to the employment of obtainable antifungal medicine together
with nephrotoxicity, hepatotoxicity and neurotoxicity [219]. Moreover,
therapeutic response is slow in disorder patients. Fluconazole is taken
into account to be one of the safest antifungals used in the treatment of
plant infections however the fungistatic nature and therefore the de-
velopment of resistance in fungi have restricted the utilization of flu-
conazole [220,221]. Resistance of human pathogens to antifungal
8
Microbial Pathogenesis 134 (2019) 103580
medicine and toxicity-connected issues have resulted the desire for
novel anti-mycotic agents with a broad spectrum of actions and fewer
dose limiting adverse reactions. Many in-vitro and in-vivo studies ex-
ploitation plant merchandise historically employed in ethnomedicine
have shown promising antifungal activity with none adverse reaction,
particularly once plant essential oils area unit used, in keeping with
World Health Organization, medicative plants would be the most ef-
fective supply of a range of medicine [222].
a. Ferulago capillaris: The Apiaceae family includes a high variety
of aromatic plants that are well-known to possess antimicrobial prop-
erties, significantly nice one to their essential oils contents. Being a
perennial genus of the Apiaceae family, the genus Ferulago is portrayed
by forty species round the world. These species are employed in drugs
for sedative, tonic, organic process and anti-parasitic effects and for the
treatment of ulcers, snakebites, haemorrhoids, headache and diseases of
the spleen [223,224]. The antimicrobial activity has previously been
rumored for a few Ferulago species, such as F. thyrsiflora, F . Sylvatica, F.
nodosa, F. bernardii, F. longistylis and F. angulata subsp. Carduchorum
[225]. Pinto et al. [226] rumored the antifungal activity of Ferulago
capillaris oil against fungus Cryptococcus fungus genus and dermatophyte
species. The target of their study was to evaluate the composition, an-
tifungal activity and mechanism of action of the EO of Ferulago capillaris
and it's main parts, terpene and α-pinene, against clinically relevant
yeasts and moulds. The study found that the MIC values of F. capillaris
oil against all the tested organisms square measure nearly adequate to
the minimum final concentration (MLC) values suggesting that the
antifungal activity of this oil can be antifungal instead of fungistatic.
The main parts of the oil, namely, α-pinene and terpene, were ad-
ditionally investigated against all the tested microorganisms. Terpene
was slightly less active than α-pinene, significantly against dermato-
phyte and Cryptococcus, and has same activity against the bulk of fungus
strains and fungus genus. Each of the isolated compounds were less
active than the oil, apart from dermatophytes, with α-pinene being a
more active. As a result, it can be understood that the oil from F. ca-
pillaris possesses a promising efficiency to be objected for additional
S. Tariq, et al. Microbial Pathogenesis 134 (2019) 103580

Table 5
(Growth inhibition (mm)͙∗ of Trichophyton spp. On sabouraud's agar plates:
FUNGI.
E.O Trichophyton erinacei T.mentagrophytes T.rubrum T.schoenleinii T.soudanense T.tonsurans

C.Atlantia
1 7.3 ± 0.76f 9.2 ± 0.29e 1.0 ± 0.00a 7.3 ± 0.58e 8.7 ± 0.58d 7.2 ± o.76d
2 3.3 ± 0.76c 5.3 ± 0.29c 0.7 ± 0.29a 4.3 ± 1.15c 5.8 ± 0.76c 3.8 ± 0.29b
C.bergamia
I 8.0 ± 0.50f 6.0 ± 1.00c 8.7 ± .76e 3.3 ± 0.58b 14.3 ± 1.53 3.3 ± 0.58
II 5.0 ± 2.00e 2.8 ± 0.29b 6.0 ± 1.00d 2.5 ± 1.50a 7.8 ± 0.29 2.3 ± 0.58a
F.Communis
I 2.8 ± 0.29b 5.8 ± 0.29c 3.8 ± 0.29c 6.5 ± 1.32 5.2 ± 0.29b 4.7 ± 0.58
II 1.7 ± 0.29b 2.7 ± 0.58b 11.0 ± 0.58 3.3 ± 0.29b 2.8 ± 0.29 2.2 ± 0.29a
M.alternifolia
I 13.0 ± 0.58 8.7 ± 0.58e 9.3 ± 0.58e 16.0 ± 1.15f 23.3 ± 1.53 5.2 ± 0.29c
II 4.8 ± 0.29e 5.3 ± 0.29c 6.7 ± 1.15e 8.3 ± 0.58 11.7 ± 0.29e 3.5 ± 0.87b
C.Citratus
I > 39.0 > 39.0 > 39.0 > 39.0 > 39.0 > 39.0
II > 39.0 > 39.0 > 39.0 > 39.0 > 39.0 > 39.0
I
II 25.0 ± 1.53 21.0 ± 2.08 15.0 ± 2.00 9.8 0.58 > 39.0 10.0 ± 0.29f
II 18.0 ± 0.00 13.0 ± 1.53 9.0 ± 2.00e 6.2 1.15d > 39.0 5.8 ± 0.76c
III 0.0 ± 0.00a 0.0 ± 0.00a 0.0 ± 0.00a 0.0 ± 0.00a 0.0 + 0.00a 0.0 ± 0.00a
L.angustifolia
I > 39.0 > 39.0 > 39.0 > 39.0 > 39.0 > 39.0
II > 39.0 > 39.0 > 39.0 > 39.0 > 39.0 > 39.0
R.Officinals
I > 39.0 > 39.0 > 39.0 > 39.0 > 39.0 31.0 ± 0.58
II > 39.0 > 39.0 > 39.0 > 39.0 > 39.0 17.0 ± 0.00
Benzoic Acid
III 1.3 ± 0.29a 2.5 ± 0.50b 5.7 ± 0.58d 2.8 ± 0.29b 6.3 ± 0.58c 5.8 ± 0.29c
P.graveolens
I > 39.0 > 39.0 > 39.0 > 39.0 > 39.0 > 39.0
II > 39.0 > 39.0 > 39.0 > 39.0 > 39.0 > 39.0
III 4.0 ± 0.50 6.8 ± 1.04d 4.3 ± 1.04d 10.5 ± 1.32 8.7 ± 1.26 9.5 ± 0.50f
Geraniol
III 4.0 ± 0.00d 6.2 ± 0.76c 3.7 ± 0.29c 4.7 ± 0.58c 8.8 ± 0.29d 9.3 ± 0.58f
Thymol
III 8.8 ± 0.76f 7.3 ± 0.58d 9.3 ± 0.58e 9.3 ± 0.58 6.3 ± 1.53c 5.8 ± 0.29c
T.Vulgaris
I > 39.0 > 39.0 > 39.0 > 39.0 > 39.0 > 39.0
II > 39.0 > 39.0 > 39.0 > 39.0 > 39.0 > 39.0
III 4.5 ± 0.87 7.8 ± 1.04 4.7 ± 0.58d 10.8 ± 1.26 11.3 ± 0.64e 8.2 ± 1.04e

investigation associate degreed development as an antifungal drug,
significantly for the treatment of fungal infection, candidiasis and
dermatophytes.
b. Curcuma longa: Curcuma longa or turmeric, a stalk nonwoody
perennial plant of the Zingiberaceae, monocot family [227]. A perennial
herb with thick and ellipsoidovate rootstock with orange cortex within.
Turmeric have been utilised in Bharat for thousands of years and may
consists a major part of Ayurvedic drugs [228]. It was Ist used as a dye
and then later for its medicative properties. Medicative uses of the
rhizomes arise from oil as a carminative and antifungal activity and
yellow curcuminoids as antioxidative and anti inflammatory properties.
In addition, it provides proof for the ethnopharmacological use of this
medical plant to treat skin diseases, particularly mycosis.
c. Eugenia caryophyllata: Unremarkably called clove, is an aro-
matic tree native of Indonesia and used as a spice in cuisines in several
components of the plane. Cloves are employed in Indian Ayurvedic
drugs, Chinese drugs, and western herbalism and medical specialty.
Cloves are afore mentioned to be a natural anthelmintic. Applied to a
cavity in a very decayed tooth, it conjointly relieves aching [229]. The
dicot genus is one among seventy five genera that's native within the
tropics, notably in tropical America and plants of this family are le-
gendary to be made of volatile oils that are reportable for people's
healthful importance. Dicot genus has properties like anti-in-
flammatory, analgesic, antipyretic, antifungal, and employed in peptic
ulceration treatment [230–232].
Gayoso et al. [233] reported antifungal impact of dicot genus con-
sisting compounds cariophyllata oil and eugenol. They evaluated the
antifungal activity of dicot genus cariophyllata oil and eugenol, its
major constituent, on plant life strains isolated from onychomycosis.
Microorganisms used were candida, fungus tropicalis, fungus krusei, Tri-
cophyton rubrum, Tricophyton mentagrophytes and Geotrichum candidum.
These microorganisms were isolated and known from clinical samples
collected from patients. Their study found that E. cariophyllata oil (MIC
2%) and eugenol (MIC 4%) are significantly active against the mould
and yeast strains isolated from onychomycosis. These plant merchandise
will so be enclosed in pharmaceutical formulations to combat the
etiological agents of onychomycosis.
Gynecologic infections are the most common reasons today why
women visits to a doctor and raise to be treated [234] in recent years,
the severity and incidence of flora infections have increased sig-
nificantly. One amongst the diseases caused by duct endogenous flora is
mycosis. It's calculable that seventy fifth of girls expertise vulvar-va-
ginal mycosis a minimum of once in their period. Virtually forty fifth of
girls are full of this infection doubly per annum. Unexpectedly, only a
tiny low range of girls are full of chronic continual infections. C. albi-
cans is that the accountable germ in 85–90% of duct yeast infections
[235]. Early designation and applicable treatment of vaginitis are ne-
cessary, as a result of failure to timely and appropriately treatment to
these infections ends up in serious complications such as pelvic in-
flammatory disease, infertility, chronic pelvic pain, premature birth,
and also the dangers of HIV infection [236]. Since the amount of pa-
tients with AIDS and unwellness and also the use of antibiotics increases
everyday and given that this fungal disease incorporates a high pre-
valence during this patient population, it's necessary to search out a

9
S. Tariq, et al. Microbial Pathogenesis 134 (2019) 103580

Table 6
Some of the plant families bearing compounds with antifungal activities.
Plant Family Botanical Name Compounds present References

1.Anacardiaceae Pistacia lentiscus α-Terpineol, [129]

Terpineol
2.Amaranthaceae Chenopodium ambrosioides m-cymene,myrtenol [130]
3.Asteraceae Arnica longifolia Camphor,1,8-cineole [131]
Aster hesperius Carvacrol,α-bisabolol [131]
Baccharis latifolia Hexadecanoic acid, Carvacrol [132]
Chrysothamnus nauseosus Camphor,α-and β-pinene,lyratyl acetate [131]
Elephantopus Spicatus β- Phellandrene,β-pinene [133]
Eupatorium Semialatum δ-elemene, farnesene, α-curcumene,selina-4,7(11)-diene,β-bisabolene [133]
Spilanthes Americana Piperitone, piperitenone [134]
4.Apiaceae Crithmum maritimum Dillapiole, γ-Terpinene,Sabinene,Thymol-methylether,β-phellandrene [135]
Daueus carota subsp.carota Sardinia:β-bisabolene,11-α-[H]-himachal-4-en-1-β-ol Portugal geranyl acetate,α-pinene [136,137]
Disticchoseelinum tenuifolium Myacene, Limonene [138]
Daucus carota subsp.halophilus Flowering umbels:Sabinene α- pinene, limonene;Ripe umbels elemicin, Sabinene [139]
Eryngium duriaei subsp.Juresianum Αneocallitropsene,Isocaryophyllen-14-al, [139]
14-hydroxy-β-caryophyllen,caryophyllene oxide,E-β-Caryophyllene
Ferula hermonis α-pinene,α-bisabolol,3,5-nonadiyne [140]
Trachyspermum ammi Thymol,P-cymene, γ-Terpinene, β-pinene,Terpinen-4-ol [141]
Coriandrum Sativum Linalool, geraniol [142]
Pimpinella anisum Trans-anethole [143]
Foeniculum graveolens Anethol, Fenchone [144]
5.Euphorbiaceae Croton cajucara Linalool [145]
6.Gentianaceae Gentiana asclepiadea Xanthones [146]
7.Hypericaceae Hypericum perforatum Terpinen-4-ol [147]
8.Labiatae Hyptis suaveolens Sabinone, Terpinolene.1,8-cineole [148]
9.Lauraceae Aniba rosaedora Linalool [149]
Laurus.nobilis 1,8-cineole [149]
Sassafras albidum Safrote [149]
Cinnamomum Zeylanicum Trans-cinnamaldehyde [149]
10.Piperaceae Piper barberi 1,8 ceneole α-pinene,eugenol isomer, camphor [150]
11.Ranunculacee Nigella Sativa Nigellone [151]

Table 7
Principle effects of essential oils and/or their components on various fungi.
Mecahnism of action Mechanism of action
Oils/compounds Oils/compounds

Antifungal Melaleuca alternifolia [156], Matricaria ricutita, Lavandula
angustifolia [157], Illicium verum, Hyssopus officinalis,
Hedychium spicatum [158], Eucalyptus [159],
Cymbopogon nardus [160], Curcuma longa [161], Coriandrum
sativum, Commiphora myrrha, Citrus [162], Cicuta virosa
[163], Cananga odorata, and Calamintha nepeta [164]
Effect on membrane/cell benzyl benzoate [168], anethole [169], Thymus [170,171],
Syzygium aromaticum [172], Salvia sclarea [173] Origanum,
Ocimum basilicum, Mentha piperita, Melaleuca alternifolia
[174], Litsea cubeba, Juniperus communis, Coriandrum sativum
[175], Coriaria nepalensis [176], and Citrus
Effect on cell growthand Thymol, terpinene-4-ol, γ-terpinene, α-terpinene, citronellal
morphology [182], p-cymene, 1,8-cineole, α-pinene, carvacrol, Thymus
spp., and Eucalyptus
Action of fungal tetraterpenoid [184], Lupeol, Origanum majorana, Origanum
mitochondria compactum [185], Hedychium spicatum, Commiphora myrrha,
Coriandrum sativum, Cinnamomum camphora, Cananga
odorata, Artemisia herba alba, and Anethumgraveolens [186]
Synergistic/antagonistic Rosa damascena [190], Pelargonium graveolens, Origanum
heracleoticum, Ocimum basilicum, Myrthus, Melaleuca
alternifolia, Matricaria recutita, Lavandula angustifolia, Illicium
verum, Eucalyptus, Cymbopogon nardus, Coriandrum sativum,
and Citrus
Inhibition of biofilm Myrtus communis, Ocimum [192,193], Piper claussenianum
development [194], Rosmarinus officinalis [195], Mentha Piper [196,197],
Melaleuca alternifolia [198,199], Litsea, Laurus nobilis [200],
Eucalyptus, Cymbopogon [201], Cytrus [202], Crotoncajucara
[203,204], and Coriandrum sativum [205]
Anti quorum sensing Origanum, Salvia sclarea, limonene, linalool, α-pinene,
terpinene-4-ol, Citrus [209], Juniperus communis, Mentha
piperita [209]
Antifungal Carvacrol [165], Viola odorata, Saturjeia hortensis,
Thymus vulgaris [166], Syzygium aromaticum,
Salvia officinalis,
Piper nigrum, Pelargonium graveolens, Origanum,
Ocimum basilicum, Myrthaceae [167], Myristica fragrans, and
Melissa officinalis
Effect on membrane/ 1,8-cineole, carvacrol [177], cinnamaldehyde [178], p-
wall cymene, citral, citronellal, eugenol, limonene, linalool,
linalyl acetate, α-pinene, α-terpinene, terpinene-4-ol, and
thymol [179–181]
Inhibition of efflux thymol [183] carvacrol, Citrus, Eucalyptus, Melaleuca
pump alternifolia, Mentha, Ocimum basilicum, Origanum vulgare,
and Thymus vulgaris
ROS production Zatharia multiflora [187], carvacrol, p- cymene, farnesol
Anti-nitric oxide [188], and thymol [189]
Synergistic/ Thymol, linalyl acetate, linalool, eugenol, Citronellal, citral,
antagonistic 1,8-cineole, carvacrol [191], benzyl benzoate, Viola odorata,
Thymus vulgaris, and Satureja hortensis
Inhibition of biofilm Syzygium aromaticum, ρ-cymene, 1-8-cineole, linalool,
development terpinolene, α-terpineol, terpinen-4-ol eucarobustol E
[206,207], eugenol [208], α-terpinene, and γ-terpinene
Effects on microtoxins Cinnamomum [210,211] Origanum vulgare, Cymbopogon
synthesis/production [211] Cider, Citrus, Eucalyptus, Mentha, Rosmarinus
officinalis, Thymus, Ocimum sanctum [212], Satureja
hortensis [213], Zataria multiflora [213] 2,3-
dideoxyglucosides, eugenol [214,215]

10
S. Tariq, et al.
secure and additionally a correct treatment. Oily essence has histori-
cally been employed in the treatment and also the bar of varied diseases
and infections [237]. In the present era, a giant trend in treatment in
America has occurred which is the use of complementary and medicine.
It includes a variety of treatments from acupuncture and prayer con-
ferences and, a lot of significantly, the utilization of seasoner medicines
that embody the normal herbs and native plants from China, India,
Africa, Europe, and America. In addition as being a natural product, has
verified that it's a comparatively safe medical aid. So, this medicine has
been emphasised due to the restricted facet effects and low price for its
application in chronic diseases medical aid and bar [238]. Lavender
(Lavandula angustifolia), a kind of medicinal herb, will be effectively
employed in the treatment of duct discharges. Lavender oil is usually
employed in traditional drugs [239] and its impact on reducing the
number of Candida albicans fungus has been shown in in-vitro [240]
and clinical studies [241]. On the opposite hand, one in all the standard
chemical medicine to treat duct mycosis is clotrimazole. Since this
chemical drug has side effects, corresponding to multiplied liver en-
zymes, painful voiding, and depression (due to general absorption of
the drug), and complications corresponding to irritation and feeling of
tingling or dermatitis [242], and because of the increasing resistance to
antifungal medicine [243], during this study, the result of lavender
(Lavandula angustifolia) and clotrimazole on the duct mycosis growth, in
vitro, was judged. With the increase in drug-resistant fungal treatment
in recent years and adverse effects of antifungal agents, the requirement
for kind excellent new medicine has become necessary. Studies on the
antifungal effects of lavender were totally different. Lavender had a quite
weak restrictive result of flora in some studies, but, in most cases, its
antifungal result was considerably positive [244]. Devkatte et al. [245]
in an exceedingly study on flavouring oils as potential inhibitors of
Candida albicans growth aimed to determine the minimum inhibitory
concentration and minimum agent concentration of thirty eight fla-
vouring oil essences. They reported that cinnamon oil was the best and
had the impact of the fungicide concentration of 0.03% against four
fungus strains. Most oils were placed in moderate cluster and nine oils
were the smallest amount effective during which the lavender was
thought of within the least effective cluster. Mahboubi et al. reportable
that lavender has moderate antifungal activity whereas alternative es-
sential oils, like thyme and herb, have strong antifungal effects. Mah-
boubi showed that some essential oils were nearest against fungus
strains [246,247]. Basically, lavender oil incorporates a long history of
medicinal use in Traditional Chinese Medicine (TCM) [248] and has
antimicrobial activity against fungi and bacterium [244,249]. Many
mixtures like to linalyl acetate, linalool, saturated fatty acid, and pro-
panoic acid are concerned during this activity. It had been shown in an
exceedingly similar study that the volatile oil of Lavandula officinalis
and its main parts, essential oil and linalyl acetate, will kill or inhibit
the expansion of fungi. Life threatening infections have increased
worldwide in disorder patients in developing countries and are getting
a very important reason behind illness and death [250]. Unhealthful
organism like yeast from fungus|fungus genus are recognized as a sig-
nificant agent of hospital nonheritable infections in humans though
emergence of infections due to non albicans Candida are represented in
recent times [251,252]. Another yeast species like Cryptococcus and
Tricosporon may cause serious diseases in disorder people [253]. Man-
agement of yeast infections will vary and area unit supported the
anatomic location of the infection, the patient's underlying sickness,
immune standing, and patients risk issue for infection, the particular
species accountable for infection and also the condition of the species to
specific antifungal medicine [254]. Among antifungal agents Azole
medicine and their derivatives still dominate as agents of selection for
the treatment of those infections, either as relevant application or oral
dosage forms. These medicines though terribly wide acclaimed for their
activity, may cause various side effects [255,256]. Drug like Flucona-
zole is fungistatic in nature and there's emergence of Fluconazole re-
sistance among clinical isolates of Candida albicans [257]. Thus there is
11
Microbial Pathogenesis 134 (2019) 103580
a need for therapeutic alternatives against yeast infections that should
be effective with lesser facet effects [258]. Varied essential plant oils
are tested for in vitro growth of C. albicans [259]. However, none of the
study demonstrates their activity on different yeast species. Thus pre-
sent study was done to understand the activity of essential plant oils
against varied yeast species isolated from clinical samples. ICU patients
were selected because they are prone to develop fungal infections.
2.4. Antiviral properties
From literature survey, it is explicit that several essential oils have
antiviral activities against many RNA and DNA viruses, such as type 1
and type 2 herpes simplex virus (HSV-1 and HSV-2), dengue virus type 2,
influenza virusadeno virus type 3, poliovirus, Junin virus, and coxsack-
ievirus B1 [260]. Bioactive pure natural compounds and Plant-based
products are a new source of antiviral drugs, as natural bioactive pro-
ducts have genetically high chemical distinctiveness. Globally, viral
diseases are so far a main concern for human well-being. So far, only a
finite count of drugs are active against several viruses, which has
evoked need for the research to find novel antiviral lead molecules. The
major constituents of MAPs along with their antiviral activities are
listed in Table 8.
The strong antiviral activities are exhibited by clove and oregano
EO's against many non enveloped RNA and DNA viruses such as polio
virus, coxsackie virus B1 and adeno virus type 3, [268,269]. Melaleuca
alternqolia EO's shows antiviral activityagainst Tobacco Mosaic Virus
(TMV) and is quite active in reducing lesion number of Nicotiana glu-
tinosa to an appreciable extent for ten days post inoculation [270].
Essential oils and the monoterpene components of thyme, eucalyptus
and tea tree possess antiviral activity against herpes simplex virus type-
1 (HSV-1) in vitro. These EO's can inhibit HSV by more than 96% and
the monoterpenes inhibited HSV by about more than 80% [271]. The
essential oils of several plant sources and their antiherpes activity as
well as several constituents of essential oils has been established earlier
[272]. The antiherpes activity of Australian tea tree oil and eucalyptus
oil [273], manuka oil [274] and thyme oil [275] have been reported
previously. Some sesquiterpenes [276] triterpenes [277] and phenyl-
propanes [278,279] have been certified for their antiviral activity
against different herpesviruses and rhinovirus. There are only a few
research articles which describe the inhibition of viral multipilication
by monoterpenes, and they have not been analysed systematically for
their antiviral efficiency. e.g. isoborneol as a potent inhibitor of HSV
[280] consequently there is little information about terpenes con-
cerning the inhibition of the viral multipilication cycle and their mode
of antiviral activity. The multipilication ability of virus HSV-1 can be
repressed by many essential oils in in-vitro experimental conditions
[281]. The main cause of some common viral infections in humans,
such as herpetic encephalitis, herpetic keratitis, neo natal herpes and
mucocutaneous herpes infections is HSV-1. Research on the essential
oils of Glechon marifolia, Artemisia arborescens and Glechon spathu-
lata found that they potentially suppressed HSV-1 [282]. Citral and
citronellal, the major components of Melissa officinalis essential oils
can inhibit the replication of HSV-2 [283]. Similarly, the antiherpes
properties of thyme oil, Australian tea tree oil and eucalyptus oil have
been reported previously [284]. ??-Caryophyllene, which is present in
essential oils of many medicinal plants, is regarded asthe best anti viral
agent [285]. Similarly, several sesquiterpenes and phenylpropanoids
including eugenol, trans-anethole, ??-eudesmol, farnesol and ??-car-
yophyllene possess antiviral properties against HSV [285]. Likewise,
eugenol, possess virucidal activity against human herpes virus
[286,287]. Some sesquiterpenes and triterpenes also possess antiviral
activity against several rhinovirus and herpesviruses [288]. Garcıaet.al
[289] reported the antiviral activity of Artemisia douglasiana and Eu-
patorium patens essential oils against the dengue virus. In addition,
Lippia turbinate and Lippia junelliana essential oils possessed activity
against the Junin virus. Anti-influenza A (H2N2) activity was exhibited
S. Tariq, et al. Microbial Pathogenesis 134 (2019) 103580

Table 8
Antiviral activity of essential oils against human pathogens.
Chemical compounds Inhibited microorganism Plant Part used

Thymol, ??-pinene, p-cymene, limonene Japanese encephalitis virus(JEV) [261] Trachyspermum ammi leaves
Patchoulol, ??-guaieno; gurjunene-??,??-guaiene, Influenza A (H2N2) virus [262] Pogostemon cablin leaves
aromadendrene,
??-patchoulene
Linalool, eugenol HSV-1 Ocimum campechianum leaves
??-Pinene, estragole HSV-1 Minthostachys mollis leaves
Myrcene, linalool, camphor, citronellal, HSV-2, avianinfluenza virus(AIV)subtypeH9N2 [11] Melissa officinalis leaves
??-caryophyllene,
caryophylleneoxide, citral
Germacrene-D HSV-1 Lepechinia salviifolia leaves
??-Phellandrene, p-cymene, HSV-1 [263] Hyptis mutabilis leaves
E-caryophyllene
??-Caryophyllene, bicyclogermacrene HSV-1 [264] Glechon marifolia leaves
??-Caryophyllene, bicyclogermacrene HSV-1 [264] Glechon spathulata leaves
2,5,5-Trimethyl-3,6- Aerial parts heptadien-2-ol, eucalyptol, ORF virus (aparapoxvirus) [265] Achilleafragrantissima Aerial parts
artemisia alcohol, thujone
Gurjunene, eudesmol, muurolene Avian influenza A virus (H5N1) [266], Fortunella margarita leaves
??-Thujone, linalool, myrcene, carvacrol Herpessimplex virus type 1 (HSV-1) [267] Artemisia arborescens Aerial parts

by the essential oil compounds of Pogostemoncablin [290] and the
antiviral property of the essential oils obtained from fruits and leaves of
Fortune llamargarita exhibited potential activity against avian influ-
enza virus (H5N1) [291]. Roy et al. [292] reported the potential anti-
viral activity of Trachyspermum oil against Japaneseencephalitis virus
(JEV). Similarly, Zeedan et al. [293] reported the antiviral activity of
Achillea fragrantissima against the ORF virus (aparapox virus). Recently,
Pour ghan bari et al. [294] evaluated invitro antiviral activity of M.
officinalis (lemon balm) essential oil and noseltamivir and their sy-
nergistic effect on avian influenza virus (AIV) subtype H9N2. They
found that various quantities of lemon balm essential oil decreased
influenza virus replication. However, it had increased efficacy when co
administered with the antiviral agento seltamivir. Essential oils ob-
tained from Colombian MAPs such as Lepechinia salviifolia, Minthos-
tachysmollis, Lepechinia vulcanicola, Hyptismutabilis, and Ocimum cam-
pechianum were reported to possess antiviral activity against human
herpes viruses types 1 and 2 [295]. They also reported that these es-
sential oils inhibit viral activity during their early stages of infection.
Thus, plant based essential oils could be used as antiviral agents against
several viral diseases in humans and have the potential to be used as
alternatives to synthetic antiviral drugs.
The volatile oil of Myrtaceaen species, specifically Eucalyptus glo-
bulus Labill. was investigated by Cermelli et al. and its effects on re-
spiratory bacteria and viruses assessed. The activity of E. globulus vo-
latile oil determined for one hundred twenty isolates of Streptococcus
pyogenes, twenty isolates of S. pneumoniae, forty isolates of S. agalactiae,
twenty isolates of S. aureus, 40 isolates of Haemophilus influenzae, thirty
isolates of H. parainfluenzae, ten isolates of enteric bacteria pneumo-
niae, ten isolates of Stenotrophomonas maltophilia and 2 viruses, a strain
of animal virus and a strain of infectious disease virus. The bactericide
activity was tested by the Kirby-Bauer paper technique, with minimum
antiseptic concentration and minimum repressing concentration. The
Kirby-Bauer paper technique, also called the agar diffusion test, is used
for measuring the effectof an antimicrobial agent against bacteria
grown in culture. By victimisation, the MTT check the toxicity was
evaluated on viro cells. The most influenced were H. influenzae, H.
parainfluenzae, S. maltophilia and S. pneumoniae. Moreover, solely a
gentle activity on epidemic parotitis virus was found [296]. Some es-
sential oils from the dicot family, like cajeput (Melaleuca Leucadendron
L.), clove (Syzygiumaromaticum (L.) Merril & amp; Perry, kanuka
(Kunzeaericoides (A.Rich.) Joy Thomps, and manuka (Leptospermum
scoparium J.R. Förster & G. Förster) were analysed by Schnitzler et al. a
spotlight of this study was to judge the oil's toxicity in a very normal
neutral red assay (NRU). This is often a cell survival assay supported the
power of viable cells to include and bind neutral red (NR). Most non-
cytotoxic concentrations for M. Leucadendron oil and S. aromaticum oil
were determined at 0.006%. K. ericoides oil and L. scoparium oil have
lot of cytotoxic with a most non-cytotoxic concentration of 0.001%.
Moreover, it was found that manuka essential oil possesses a high ca-
pacity of agent activity against HS1 still as against drug-resistant HS1
isolates in microorganism suspension tests [297]. Also, the anti-
microorganism activity of lemon balm oil, the volatile oil of genus
Melissa officinalis and L. Lamiaceae, against enveloped herpes viruses
was investigated. It has been reported that monoterpenes and of the
essential oils from eucalyptus (Eucalyptus sp., Myrtaceae), tea tree (M.
alternifolia, Myrtaceae) and thyme (Thymus sp., Lamiaceae) and of their
major monoterpene compounds a-terpinene, g-terpinene, a-pinene, p-
cymene, terpinen-4-ol, α-terpineol, thymol, citral and 1,8-cineoleshow
antiviral activity against HSV-1 in vitro. The results showed that these
essential oils led to a discount of infective agent activityby ninety six.
The essential oils and monoterpenes revealed only moderate antiviral
effects when they were added to host cells prior to infection or after
entry of HSV into cells. All the tested medicines led to associate degree
interaction in a very dose dependent manner with HSV particles. The
study showed that among the analysed compounds monoterpene hy-
drocarbons were slightly superior to monoterpene alcohols in their
antiviral activity. The highest selectivity index was shown by α-pinene
andα-terpineol. Moreover, the mixtures of different monoterpenes, that
square measure gift in natural tea tree oil, disclosed a 10-fold higher
property index and a lower toxicity than its isolated single mono-
terpenes [298]. Garazzo et al. investigated the in vitro antiviral activity
of M. alternifolia oil (tea tree oil, TTO), Myrtaceae, and of its main
parts, terpinen-4-ol, α-terpinene, g-terpinene, p-cymene, terpinolene
and a-terpineol. The antiviral activity was tested against infantile pa-
ralysis sort one, Echo 9, Coxsackie B1, adeno sort a pair of, herpes
simplex virus and herpes simplex by five hundredth plaque reduction
assay. The anti-influenza virus assay was based on the inhibition of the
virus-induced cytopathogenicity. The results showed that Time Trade
Of or Tactical Technology Office (TTO) and a few of its compounds, e.g.
terpinen-4-ol, terpinolene and α-terpineol, possess an inhibitory effect
on influenza A/PR/8virus subtype H1N1 replication at non-cytotoxic
concentrations. Further more, TTO showed AN ID50 worth of 0.0006%
(v/v) that was abundant below its CD50 worth with 0.025% (v/v). All
the compounds showed no agent activity against infantile paralysis 1,
adeno-2, Echo9, Coxsackie B1, herpes simplex virus and herpes sim-
plex, while TTO exhibited a slight virucidal effect against HSV-1 and
HSV-2. The results of this study showed that TTO is a promising drug in
the treatment of influenza virus infection [299].

12
S. Tariq, et al.
2.4.1. Efficacy and toxicity on the development of an efficient antiviral drug
Since the discovery of IDU 50 years ago, only a few molecules have
proven to be effective and safe when used for selective antiviral
therapy. A huge breakthrough that came from the better understanding
of virus-host interaction was the inception of 9-(2-hydro-
xyethoxymethyl) guanine (Acyclovir). It was the first highly selective
antiviral drug, being a substrate for the Herpes Simplex Virus-encoded
thymidine kinase. It displayed a direct inhibitory effect against viral
replication and practically no adverse effects on the host. The
achievement of selective viral toxicity by Acyclovir and other similar
molecules were thought of as the beginning of a new therapeutic age for
a well-established, effective and safe antiviral therapy. Acyclovir is a
pro-drug, which means it has to be further metabolized in vivo before
entering the infected cell wherein further metabolism may or may not
be required to yield the active inhibitor. The key to Acyclovir's speci-
ficity is the selective phosphorylation of the acyclic guanosine nucleo-
side by the Herpes virus encoded pyrimidine deoxynucleoside kinase,
which means it would only be active on Herpes-infected cells [300].
2.5. Antibiofilm activity of essential oils
A biofilm is a complex matrix of microorganisms in which cells bind
together and attach to biotic or abiotic surface [301]. Biofilms some-
times create a sticky gel composed of polysaccharides, proteins and
other organic parts on a wet surface, found in numerous environments
as well as clinical and industrial, food process environments, and drink
distribution systems [302]. Bacteria within biofilms are more resistant
to antibiotics and chemical agents than planktonic cells in suspension
[303]. Chemical agents penetrating into the biofilm matrix area are less
effective, as a result of most of the chemicals area unit active sole ly
against unattached microorganisms. In order to penetrate and degrade
biofilms, it's necessary to change the biofilm matrix. Proscribing the
growth and development of food borne and health facility pathogens
such as staphylococci aureus and escherichia is incredibly important,
however the eradiation of these organisms is not always successful
because of their ability to form biofilms on a various range of surfaces
[304]. Interest in natural antimicrobial merchandise has multiplied in
recent years. The foremost vital and well researched compounds ori-
ginate from plants that show several medicative and antimicrobial
Properties [305] including potential activity against biofilm formation
[306]. Extracts and essential oils from a large variety of healthful plants
have attracted and inspired analysis interest. The plant extracts have
widespread application within the pharmaceutical business; as a result,
they contain varied bioactive compounds with antimicrobial properties.
For example biofilm on cucumber is eliminated by result of plant ex-
tracts (clove oil loaded with chitosan nanoparticle) are used against E.
Coli [307], Listeria monocytogenes [308], S. aureus [309] and Candida
albicans [310]. Plant compounds with robust antibacterial drug or
germicidal activity belong largely to the cluster of phytoalexins to-
gether with essential oils, [311]. Essential oils are volatile compounds
with antimicrobial properties constituting non-supportive media for the
growth of many bacteria and fungi. Several studies have reported the
antimicrobial properties of essential oils [312]. They constitute com-
plex and heterogenous mixtures of substances comprising several
structure classes with different biosynthetic origin: the main group in-
cludes terpenes (monoterpenes, sesquiterpenes) and terpenoids, to-
gether with aromatic (phenylpropanoids) and/or aliphatic compounds
[313]. Essential oils are readily isolated from plant material, exert low
toxicity in mammalians, and degrade quickly and easily in water [314].
In recent years, studies on the antibiofilm activity of essential oils have
been intensified. Antibiofilm activity of essential oils has been reported
against S. aureus by using thymoquinone, an active principle of Nigella
sativa L. seed oil [315]. Lemongrass oil (Cymbopogon flexuosus (Nees ex
Steud.) W. Watson) [316] oregano oil (Origanum vulgare L.), carvacrol
and thymol [317], oregano oil (Origanum onites L.) [318] cassia (Cin-
namomum cassia (Nees & T. Nees) J. Presl), Peru balsam (Myroxylon
13
Microbial Pathogenesis 134 (2019) 103580
balsamum (L.) Harms (L.) Harms), and red thyme (Thymus vulgaris L.)
essential oils [319] have Potential antibiofilm effect against E. coli has
been shown for tea tree, lavender, and melissa oil [320] cinnamon oil
(C. cassia (Nees & T. Nees) J. Presl) and cinnamaldehyde [321] and
eugenol and carvacrol [322].
2.6. Quorum sensing
In biology, quorum sensing is that the ability to notice and to reply
the cell population density by sequence regulation. Assemblage sensing
(QS) allows microorganism to limit the expression of specific genes to
the high cell densities at that the ensuing phenotypes are going to be
themost useful. Several species of microorganism use assemblage sen-
sing to coordinate organic phenomenon in keeping with the density of
their native population. In similar fashion, some social insects use as-
semblage sensing to work out wherever to nest. Quorum sensing (QS)
may be a communication system through that bacterium converse with
each other and better species [323]. QS is predicated on the synthesis
and perception of specific chemical signals, typically mentioned as
autoinducers, that accumulate within the growth medium throughout
microorganism growth. Once the concentration of autoinducers reaches
a threshold worth, such as a precise population density, it alters the
expression of genes. In several infective bacterium, QS completely
regulates genes answerable for virulence and biofilm formation [324].
2.6.1. Bacterial quorum sensing
Bacterial Quorum sensing (QS) may be a type of cell-to-cell com-
munication that is important to the pathogenicity of the many bac-
terium, and thus a promising target for the event of latest treatments for
microial infections. Several medicative plants possess medicinal ac-
tivity, solely many plants are shown to focus on assemblage sensing.
Many microorganism resistance to existing antibiotics in addition to the
decline in novel antibiotic development, QS inhibitors from medicative
plants may be a promising direction for brand spanking new medicinal
treatments. The aim of this review is to summarize the verified in-
formation on the anti-QS properties of medicative plants and also the
numerous mechanisms of their actions [325].
2.6.2. Role of essential oils in bacterial quorum sensing
Most QS inhibitors, both natural and artificial, belong to a family of
secondary metabolites referred to as furanones, that area unit structu-
rally lookslike AHLs and so bind to acyl-homoserine lactone (AHL) re-
ceptors. However, QS inhibitors area unit incapable of activating AHL
receptors. Asian Nation includes a wealth of ayurvedic medicines, and
Indian folks have long used ayurvedic herbs to treat contagions.
Cinnamon could be a dietary phytochemical that shows antimicrobial
properties and is especially vital as long as dietary chemicals area unit
deemed trustworthy and used routinely in everyday life. Cinnamon is
conventionally supported for treating biological process issues, as well
as nausea, vomiting, and symptom [326]. Cinnamon oil is additionally
employed in many toothpastes as associate antimicrobial substitute.
The target of this study was to analyze the power of cinnamon oil to
inhibit QS-mediated virulence factors and biofilm formation in Pseu-
domonas aeruginosa PAO1. Ferula (Ferula natural resin L.) and Dorema
(Dorema aucheriBioss.) each from Apiaceae family were tested for anti
quorum sensing (QS) activity against bacteria genus aeruginosa. Ferula
[Ferula asafetida L. (Apiaceae)] is associate non woody plant with a
good distribution in arid climates as well as Iran, Islamic State of Af-
ghanistan and Republic of India. This plant is wide accustomed alle-
viate bacterial infections in western a part of Asian country. Dorema
[Dorema aucheri Boiss. (Apiaceae)] may be a massive rosid dicot family
herb that's conjointly growing in western mountains of Asian country
and in Persian known as kandale koohi. Medicinally, the roots of D.
aucheri are used treat microbial infections. This species is the 1st herb
found to provide flavonoids that acknowledge as antimicrobial agents
[327]. Due to their use in treatment of microbial infections, it's
S. Tariq, et al.
attainable that these plants could possess anti-quorum sensing (QS)
properties. Bacterial gathering sensing (QS) could be a density depen-
dent communication system that regulates the expression of genes as
well as production of virulence factors in several pathogens. Bioactive
plant extracts inhibiting QS regulated organic phenomenon could also
be a possible candidate as antipathogenic drug. During this study anti-
QS activity of peppermint (Mentha piperita) oil was first tested. Further,
the findings of the current investigation discovered that flavouring
(PMO) at sub-Minimum repressive Concentrations (sub-MICs) power-
fully interfered with group homoserine lactone (AHL) regulated viru-
lence factors and biofilm formation in bacteria genus aeruginosa and
Aeromonas hydrophila. The results of molecular tying up analysis at-
tributed the QS repressive activity exhibited by PMO to application.
Assessment of ability of application to interfere with QS systems of
varied gram-negative pathogens comprising numerous AHL molecules
discovered that it reduced the AHL dependent production of violacein,
virulence factors, and biofilm formation indicating broad-spectrum
anti-QS activity. Mistreatment 2 escherichia biosensors, MG4/pKDT17
and pEAL08-2, we tend to additionally confirmed that application re-
strained each the las and pqs QS systems. Further, findings of the in
vivo studies with application on roundworm model Caenorhabditis ele-
gans showed considerably increased survival of the roundworm.
The essential oil of plants that are commonly used both in tradi-
tional and modern systems of medicine are anti-quorum sensing testing.
Twenty one essential oils have been screened and the result of that
indicated that cinnamon, peppermint, clove and lavender oils have
potential anti-QS activity on C. violaceum pigment production [328].
The C. Violaceum produced extent violacein which was studied in pre-
sence of clove oil using the method of Blosser and Grey 2000 [329]. At
lower concentration (2 μL), the clove oil shows no activity but at higher
concenration (20 μL), antibacterial activity was seen with anti-quorum
sensing activity. Vattem et al. (2007) described the method to assess
anti-QS nature of clove oil, against Ps. aeruginosa PAO1, the effect of
sub-MICs of clove oil on swarming motility was investigated. At 3.2%
v⁄v treatment of clove oil to be inhibitory to the growth of PAO1 [329].
GC-MS analysis revealed that the clove oil contains eugenol (74.32%)
and other compounds like α-caryophyllene (4.05%)and β-car-
yophyllene(4.92%), naphthalene, 1,2,3,5,6,8a-hexahy-dro-4,7-di-
methyl-1-(1-methylethyl) (7.04%),caryophyllene oxide (2.41%), 1,6-
Octa-diene-ol-,3,7-dimethylacetate (1.28%) and iso-caryophyllene
(5.96%). Eugenol as a major active component and well known for
antimicrobial action it was found that the anti-QS activity of clove oil is
probabablybecause of other ingredients like α-caryophyllene and β-
caryophyllene either in combination with other ingredients or alone. It
has been reported that the anti- QS of clove oil is concentration de-
pendent. Furthermore, inhibiting the swarming mortility of PAO1 by
clove oil strengthened its anti-QS behavior [330]. The essential oils
possess new antipathogenic properties that are because of its anti- QS
activity which prove to be important in rendering pathogenicity and
virulence of drug resistant bacteria in vivo.
3. Conclusions
The literature which has been published on antimicrobial activity of
essential oils makes it crystal clear that essential oils are embellished
with unique antibacterial, antifungal and antiviral properties and as
such can be utilised as antimicrobial agents. Essential oils are regarded
as GRAS (generally regarded as safe) grade chemicals by The U.S. Food
and Drug Administration (FDA) as such there are no toxicity issues
associated with them. Additionally because of their pleasant fragrance,
they can even be used in food applications.
Appendix A. Supplementary data
Supplementary data to this article can be found online at https://
doi.org/10.1016/j.micpath.2019.103580.
14
Microbial Pathogenesis 134 (2019) 103580
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