Rev Environ Health 2018; 33(1): 63–76
Review
Harriet Whiley*, Sharyn Gaskin, Tiffany Schroder and Kirstin Ross
Antifungal properties of essential oils for
improvement of indoor air quality: a review
https://doi.org/10.1515/reveh-2017-0023
Received August 1, 2017; accepted September 4, 2017; previously
published online October 27, 2017
Abstract: Concerns regarding indoor air quality, par-
ticularly the presence of fungi and moulds, are increas-
ing. The potential for essential oils to reduce, control or
remove fungi, is gaining interest as they are seen as a
“natural” alternative to synthetic chemical fungicides.
This review examines published research on essential oils
as a method of fungal control in indoor environments.
It was difficult to compare the relative performances of
essential oils due to differences in research methods and
reporting languages. In addition, there are limited stud-
ies that scale up laboratory results and assess the efficacy
of essential oils within building environments. However,
generally, there appears to be some evidence to support
the essential oils clove oil, tea tree oil, oregano, thyme and
lemon as potential antifungal agents. Essential oils from
heartwood, marjoram, cinnamon, lemon basil, caraway,
bay tree, fir, peppermint, pine, cedar leaf and manuka
were identified in at least one study as having antifungal
potential. Future studies should focus on comparing the
effectiveness of these essential oils against a large number
of fungal isolates from indoor environments. Studies will
then need to focus on translating these results into realis-
tic application methods, in actual buildings, and assess
the potential for long-term antifungal persistence.
Keywords: fungicidal; indoor air; natural; plant-derived
compound; plant extract.
*Corresponding author: Harriet Whiley, Environmental Health,
Science and Engineering, Flinders University, GPO Box 2100,
Adelaide 5000, Australia, E-mail: Harriet.Whiley@flinders.edu.au
Sharyn Gaskin: Occupational and Environmental Health, School
of Public Health, The University of Adelaide, 28 Anderson St
Thebarton, Adelaide 5031, Australia
Tiffany Schroder and Kirstin Ross: Environmental Health, Science
and Engineering, Flinders University, GPO Box 2100, Adelaide 5000,
Australia
Introduction
Indoor air quality is a public health issue of increasing
concern (1–3). One of the leading indoor air quality com-
plaints is the presence of fungi and moulds, which have
been associated with increased risk of adverse health
effects (4, 5). The most common adverse health effects
associated with fungi in indoor environments, as recently
reviewed by Nevalainen et al. (6), are various respiratory
conditions. Other potential health effects include aller-
gic responses, infection or toxigenic effects, for which
the pathophysiology is less evident (2). Of these, infec-
tion and toxicity have serious consequences but are rare
in occurrence, whereas allergic responses are commonly
observed (7). These allergic responses include rhinitis, eye
irritation, cough and aggravation of asthma, which is of
emerging significance given that the incidence of asthma
in children from developed countries is increasing (8–10).
Burr et  al. (11) conducted a randomised control trial to
explore the effect of eradicating visible mould from the
homes of asthma patients. It was found that the symp-
toms of asthma and rhinitis improved and medication
usage decreased in the patients who had indoor mould
removed, fungicide applied and a fan installed in the loft
of their homes.
Fungi and moulds have also been demonstrated to
contribute to sick building syndrome and other build-
ing related illnesses (7, 12). Sick building syndrome is
recognised as a group of symptoms (e.g. eyes, nose and
throat irritation; dry skin, headache and lethargy) that are
related to spending time in a particular building. This is
more commonly identified in offices; however, there have
been reports of sick building syndrome in schools, hos-
pitals, care homes and domestic houses (13). Research
has demonstrated that one of the key risk factors of sick
building syndrome is dampness, which promotes mould
growth (14).
To reduce the potential risk of exposure, indoor areas
with visible fungal growth must be immediately remedi-
ated (15). Early intervention for fungal contamination is
essential; otherwise, professional remediation will be
required (3). Remediation typically involves the removal
64      Whiley et al.: Indoor air and antifungal essential oils
of building material with visible mould contamination, in
conjunction with treatment of surfaces with an antifungal
product. Remediation should also include steps to prevent
moisture build-up, which enables future fungal growth
(16). An antifungal agent (otherwise known as a fungicide)
is a compound used to kill or inhibit the growth of fungi
and/or fungal spores (sporicide) (17). Antifungal agents rec-
ommended for indoor environments should be non-toxic to
humans, odourless and hypoallergenic (18). It is ideal that
the antifungal agent also provides long-term protection
from fungal regrowth, especially in humid or moist envi-
ronments that would promote fungal growth; however, in
reality, this long-term persistent protection is difficult to
achieve especially for non-toxic fungicides (19, 20).
Globally, there is increasing concern regarding syn-
thetic chemical usage and residue and the potential
human health effects of exposure (21). Consequently,
there is a push from consumers for ‘natural’ alternatives
to chemical antifungal products for use in residential
and commercial indoor environments (2, 15, 22). As such,
research on essential oils and their potential antimicrobial
capabilities has received increasing attention (23). Essen-
tial oils are complex mixtures of volatile compounds bio-
synthesised by plants, the main groups of which include
terpenes and terpenoids and aromatic and aliphatic con-
stituents, and are characterised by low molecular weight
(24). Essential oils have been widely used in medicine
and the food industry for their antimicrobial properties;
however, there are limited studies investigating their use
for the control of indoor air quality. The increased interest
in natural substances is driving the research community
to find new applications of these substances. The aim of
this review was to examine studies that have investigated
the antifungal potential of essential oils specifically as
a method for improving indoor air quality for building
occupants.
Methods
A search was conducted through the Scopus (25), Pro-
Quest (1), Science Direct (6) and Web of Science (26)
databases. The search terms included (mould OR mold
OR fungi OR fungal OR fungus) AND (“indoor air” OR
indoor OR building OR buildings) AND (“essential oil” OR
“essential oils” OR “plant-derived compound” OR “plant
extract”) AND (antifungal OR fungicidal OR fungicide OR
sporicidal OR sporicide OR anti-microbial OR biocide OR
biocidal), and the search was limited to the title, abstract
and keywords. Figure 1 presents the systematic approach
to article inclusion or exclusion. Articles were screened by
reading titles and abstracts and initially excluded if they
did not describe original research or did not examine the
fungicidal activity of essential oils. Articles were then read
in full and excluded if they described a fungal control not
relating to indoor air or buildings (e.g. articles describ-
ing the control of clinical isolates, animal, crop or food
spoilage, etc. were excluded). A total of 19  studies that
described the antifungal potential of essential oils or their
extracts for the purpose of influencing indoor air quality
were included and are summarised in Table 1.
Antifungal potential of essential oils for
the control of indoor air environments
The biggest challenge when evaluating the antifungal
potential of essential oils or their extracts (Table 1) is the
lack of a standard method for both designing experiments
and describing antifungal efficacy (43). The most com-
monly adopted screening method identified in the studies
shown in Table 1 was the disk diffusion assay. This is where
the essential oil or treatment is added to filter paper discs
and placed in the centre of agar plates containing fungal
lawn. The zone of clearing in fungal growth is measured as
an indicator of fungal inhibition. Other studies also used
modified versions of this method, including adding the
essential oil through syringe injection or placing in a well
in the centre of the agar plate (26, 36, 37). Rogawansamy
et  al. (15) used the disk diffusion assay in addition to a
method adapted from Soylu et  al. (44) to investigate the
antifungal efficiency of the treatments in the vapour
phase. Briefly, agar plates with fungal lawns are created
and filter paper containing the treatment is placed on
the inner surface of the agar plate lid, ensuring no direct
contact with the fungal lawn. Plates are sealed with para-
film and antifungal efficiency is determined by measur-
ing the zone of clearing. Another method frequently used
was the serial dilution method, where serial dilutions of
essential oils were prepared and inoculated with fungal
cultures to determine the minimum concentration that
inhibited fungal growth (25, 32, 40).
Even when the same method was used, it is difficult to
compare results as a consequence of different reporting lan-
guage used to describe antifungal efficacy. Using the disk
diffusion assay, researchers reported the results as either
diameter of zone of inhibition (15, 37) or as percentages of
growth inhibition compared with the control plates (25, 29,
33). Using the serial dilution method, Šegvić Klarić et  al.
(40) reported antifungal efficacy using the term minimum
inhibitory concentration (MIC), which is the lowest con-
centration that allowed no more than 20% fungal growth
 Whiley et al.: Indoor air and antifungal essential oils      65

Records identified through databases Scopus (25), ProQuest (1), Science direct (6)
and Web of Science (26) databases. Keyword search (mould OR mold OR fungi OR
Identification

fungal OR fungus) AND (“indoor air” OR indoor OR building OR buildings) AND

(“essential oil” OR “essential oils” OR “plant-derived compound” OR “plant extract”)
AND (antifungal OR fungicidal OR fungicide OR sporicidal OR sporicide OR anti-
microbial OR biocide OR biocidal) and the search was limited to include peer
reviewed articles written in English (n = 65)
Screening

Records after duplicates removed

(n = 44)

Records screened Records excluded

(n = 44) (n = 16)
Eligibility

Full-text articles assessed for Full-text articles or resources

eligibility excluded, with reasons
(n = 28) (n = 9)
Included

Studies included in
qualitative synthesis
(n = 19)

Figure 1: Overview of search methods and article inclusion and exclusion criteria (1, 6, 25, 26).

(determined by a reduction in the number of colonies in 10
μL of the dilution inoculated onto Sabouraud Glucose Agar
incubated at 25 °C for 7  days), and minimum fungicidal
concentration (MFC), which was the lowest concentration
of essential oil that completely inhibited the growth of the
fungi. However, this differs from the definitions used by
Stupar et  al. (32), who also used a variation of the serial
dilution method but reported the MIC as the lowest concen-
tration without visible growth (assessed using a binocular
microscope) and the MFC as the lowest concentration with
no growth after inoculation of the original inoculum. This
definition of MIC was supported by Verma et al. (45), who
reported MIC as the lowest concentration that resulted in
no growth after the incubation period.
The other challenge when evaluating the antifungal
efficacy of essential oils is that most of the studies iden-
tified in Table 1 are laboratory based and there is a lack
of in situ experiments within building environments.
This makes it difficult to translate the experimental find-
ings into ‘real-world’ recommendations. Only one study,
by Su et  al. (23), investigated the antifungal efficacy of
essential oils by evaporating essential oil in indoor rooms
and measuring the changes in air quality. Another four
studies assessed the antifungal efficacy of essential oils
against fungal growth on different types of wood surfaces
used in building construction; however, all other studies
­identified were conducted on agar plates or broth cul-
tures (36–39). There is clearly need for further research
designed to investigate the antifungal efficacy in indoor
environments (in situ) in order to validate the transla-
tion of laboratory-based outcomes. There is also a need
to investigate the potential long-term persistence of the
treatment and any optimum reapplication requirements
in order to characterise antifungal capabilities.
Clove oil
Of the essential oils identified in Table 1, clove oil has
been researched the most extensively, and there are a
Table 1: Studies investigating the antifungal activity of essential oils for the control of fungi in indoor environments.
Treatment
Extracts from heartwood (Pinus   Alternaria alternata, Fusarium   Wood specimens treated at the level of
rigida), eucalyptus leaves
(Eucalyptus camaldulensis) and globosum, Aspergillus niger
creper ginger rhizomes (Costus
speciosus)
Extracts from marjoram
(Origanum majorana), thyme
(Thymus vulgaris) and ginkgo
leaves (Ginkgo biloba)
Extracts from myrrh
(Commiphora myhrra)
  Fungal species
subglutinans, Chaetomium
and Trichoderma viride
  C. globosum (ATCC 6205),
A. niger (ATCC 9642),
Aureobasidium pullulans
(ATCC 15233), Gliocladium
virens (ATCC 9645) and
Penicillium pinophilum (ATCC
11797)
  Acremonium strictum,
Aspergillus flavus,
Aspergillus sydowii,
C. globosum, Cladosporium
cladosporioides, Cladosporium and the lowest growth inhibition (12.7%)
sphaerospermium,
Cladosporium
verrucocladosprioides,
Cochliobolus spicifer,
Drechslera biseptata,
Embellisia chlamydospora,
Eurotium amstlodami, Fusarium
semitectium, Myceliophthora
lutea, Penicillium chrysogenum,
Penicillium fellutanum,
Penicillium reticulosum, Phoma
tropica, Torula caligans,
Trichoderma psudokoningii and
Ulocladium consortiale
  Efficacy
 
2% concentration of heartwood extract
observed good inhibition to the mould
growth
  Marjoram extracts demonstrated  
excellent antifungal performance in the
laboratory experiments and when applied
to the antifungal mortars
  The antifungal efficacy of myrrh was  
highly dependent on the sensitivity of
the fungal species. The highest growth
inhibition (74.6%) was against A. strictum – Fungicidal ability tested against
was against U. consortiale
  Strengths   Weaknesses   Ref.
– Moderate transferability of   – Unknown relevance of fungi to   (27)
results to real-world application environmental isolates (stock cultures
(wood blocks were immersed in used)
the treatment prior to placing
on agar plates inoculated with
fungal species)
– Efficacy compared to DMSO as a
positive fungicidal control
– Multiple time periods assessed
– Moderate transferability of   – Unknown relevance of fungi to   (28)
results to real-world application environmental isolates (ATCC strains
(treated mortars were placed used)
on agar plates inoculated with
fungal spores. A mould growth
index of 0–4 was then used to
quantify the fungal growth.)
66      Whiley et al.: Indoor air and antifungal essential oils
– Efficacy compared to commercial
chemical antifungal agents
– Multiple time points
assessed, including long-term
performance (up to 12 months)
– Relevant fungal species used   – Limited transferability of results   (25)
(isolated from indoor air to real-world application (growth
environment) inhibition percentage calculate using
disk diffusion assay)
a large number of fungal – Multiple time points were not
isolates assessed
– Efficacy not compared to a standard/
commercially available fungicide
Table 1 (continued)
Treatment  
® ®
Cavicide and Virkon , 70%  
ethanol, vinegar (4.0%–4.2%
acetic acid) and tea tree oil
(Melaleuca alternifolia)
Clove oil (Syzygium aromaticum)   Three white-rot fungi (Trametes  50 μg/g clove essential oil had 100%
hirsuta, Schizphylhls commne,
and Pycnoporus sanguineus)
(common causes of wood rot)
Cinnamon oil (Cinnamomum  
verum) and clove oil
(S. aromaticum)
Cinnamon oil (C. verum) and  
clove oil (S. aromaticum)
Fungal species  
Aspergillus fumigatus and  
P. chrysogenum
Aspergillus sp. (WU1003),  
Trichothecium sp. (WU1004),
Trametes sp. (WU1005) and
Gloeophylum sp. (WU1006)
Aspergillus sp. WU1003 and  
Trichothecium sp. WU1004
Efficacy   Strengths   Weaknesses   Ref.
Tea tree oil demonstrated the greatest   –R
 elevant fungal species used   – Limited transferability of results   (15)
inhibitory effect on the growth of both (isolated from indoor air to real-world application (growth
fungi, followed by Cavicide® and Virkon®. environment) inhibition zones were measured
Vinegar only inhibited P. chrysogenum –E
 fficacy compared to using disk diffusion assay on agar
and 70% ethanol had no inhibitory effect commercially available and antifungal activity in vapour
fungicides phase was assessed using by placing
a treated filter paper disk on the
inside lid of an agar plate containing
a fungal lawn)
– Multiple time periods were not
assessed for the disk diffusion method
    – Limited transferability of results to   (29)
mortality to Reticulitermes chinensis after real-world application (antifungal
testing for 5 days index was calculated using the agar
dilution method)
– Unknown relevance of fungi to
environmental isolates (stock cultures
used)
– Efficacy not compared to a
commercially available fungicide
– Multiple time points were not examined
3% concentration of cinnamon and clove   – Moderate transferability of   – Unknown relevance of fungi to   (30)
oil gave complete inhibition against results to real-world application environmental isolates (stock cultures
Aspergillus sp. and Trichothecium (particleboards were treated used)
sp. on rubberwood particleboard for and then dipped into the – Efficacy was not compared to a
9 weeks. Particleboards treated with mould spore inoculum and commercially available fungicidal
clove or cinnamon oil were found to have then incubated in a humid control
reduced mass loss from Trametes sp. and environment)
Gloeophylum sp. compared to the controls. – Results were measured over time
The percentage loss of mass decreased with
increasing levels of clove and cinnamon oils
Dip treatment in cinnamon oil and clove oil   – Moderate transferability of   – Unknown relevance of fungi to   (31)
at a concentration of 0.63% was capable results to real-world application environmental isolates (stock cultures
of providing complete protection for at (particleboards were treated used)
least 8 and 5 weeks, respectively. Contents and then dipped into the – Efficacy was not compared to a
of cinnamaldehyde and eugenol in the mould spore inoculum and commercially available fungicidal
particleboards, the main antifungal agents then incubated in a humid control
in cinnamon oil and clove oil, respectively, environment)
largely declined within the first 4 weeks of – Multiple time points assessed
incubation, which could explain the time
Whiley et al.: Indoor air and antifungal essential oils      67
limit of the fungicidal activity
Table 1 (continued)
Treatment  
Oregano (Origanum vulgare)
essential oil
Essential oils from caraway
(Carum carvi L.), bitter orange
(Citrus aurantium L.), bergamot
orange (Citrus bergamia Risso
& Poit), coriander (Coriandrum
sativum L.), common juniper
(Juniperus communis L.), English
lavender (Lavandula angustifolia
Mill.) corn mint (Mentha arvensis
L.), pennyroyal (Mentha pulegium
L.), basil (Ocimum basilicum L.),
lemon basil (Ocimum citriodorum
Vis), marjoram (O. majorana L.),
oregano (O. vulgare L.), bay tree
(Pimenta racemosa (Mill.) J.W.
Moore), rosmary (Rosmarinus
officinalis L.), common sage
(Salvia officinalis L.), sage (Salvia
sterea L.), tansy (Tanacetum
vulgare L.), thyme (Thymus
satureoides Coss. & Balansa.,
T. vulgaris L.) and ginger (Zingiber
cassumunar Roxb.)
Siberian fir (Abies sibirica L.),
common caraway (Carum carvi
L.), peppermint (Mentha piperita were A. versicolor, A. niger,
L.), willow-leaved gum tree
(Eucalyptus globulus Labill.),
lemon thyme (Thymus pulegioides C. cladosporioides,
L.), clove tree (S. aromaticum
(Linn.) Merril & Perry) and
bergamot orange (C. bergamia
Risso & Poit)
Fungal species  
  A. fumigatus,  
Aspergillus nidulans,
Aspergillus versicolor and one however, it was not as effective as biocide
Penicillium species
  A. alternata, Stachybotrys  
chartarum, C. cladosporioides The group of most effective essential
and A. niger
  The dominant species isolated   The highest fungicidal activity was
from walls and air samples
A. fumigatus, Cladosporium
sphaerospermum,
P. chrysogenum,
P. aurantiogriseum,
P. simplicissimum and
Efficacy  
Oregano demonstrated strong antifungal   – Relevant fungal species used
activity against all fungal isolates tested;
and benzalkonium chloride
High antifungal activity  
oils including coriander, lemon basil,
oregano, caraway, bay tree and thyme
achieved high inhibition levels up to
100% across the entire spectrum of target
pathogenic fungi
Moderate antifungal activity:
English lavender, pennyroyal, corn
mint and sage exceeded 50% inhibitory
effect boundary level in all target
species except A. niger. Marjoram also
did not exceed 50% inhibitory effect in
C. cladosporioides
Low antifungal activity:
Bitter orange, bergamot orange, common
juniper,, basil, common sage, tansy and
ginger all failed to reach 50% antifungal
inhibitive effect in a single target fungi
 
demonstrated by clove oil. All cultures
were affected and the fungicidal zones
ranged from 20 to 50.5 mm. This
fungicidal activity was comparable to the – Fungicidal ability tested against
most effective disinfectant (Biosheen
20.9–57.5 mm). Next, fir oil was also
effective against all fungi tested, but its
fungicidal impact was less than that of
clove oil
Strengths  
 
(isolated from indoor air
environment)
– Efficacy compared to commercial
biocide (benzalkonium chloride)
– Multiple exposure times were
assessed
– Large number of essential oils   – Unknown relevance of fungi to
tested
– Relevant fungal species used  
(isolated from indoor air and
surfaces of water damaged
buildings)
a large number of fungal
isolates
– Efficacy compared to commercial
disinfectants
Weaknesses   Ref.
– Limited transferability of results to   (32)
real-world application [antifungal
activity was assessed using the
suspension-neutralisation method on
malt extract agar and the agar micro-
dilution technique (results reported
as MIC and MFC)]
  (33)
environmental isolates (stock cultures
used)
– Limited transferability of results
to real-world application (agar
microdilution method on potato
dextrose agar. Antifungal activity was
reported as percentage of growth
inhibition compared to control.)
68      Whiley et al.: Indoor air and antifungal essential oils
– Multiple time points were not
assessed
– Efficacy not compared to commercial
fungicide
– Limited transferability of results to   (34)
real-world application (disk diffusion
assay with results reported as
diameter of fungal inhibition zones)
– Incubation time was describes as
3–5 days. The effect of time was not
assessed
Table 1 (continued)
Treatment  
 
Crude extract of glycoalkaloids  
from nightshade (Solanaceae)
plants
Essential oils from black peper   A. niger and Geotrichum
(Piper nigrum Linn.), castor oil
(Ricinus communis Linn.), cedar
(Cedrus deodara (Roxb.) Loud.),
clove (S. aromaticum Linn.),
eucalyptus (Merrill & Perry
E. globulus Labill.), bitter orange
(C. aurantium Linn.), lemon
(Citrus limon (Linn). Burm. f),
olive (Olea europaea Linn.), and
peppermint (M. piperita Linn.)
Thujopsene (found in the  
essential oil of a variety of
conifers)
Fungal species
Ulocladium chartarum.
In addition, the fungus
Penicillium digitatum was
found spreading on the wall in
one case
Aspergillus, Penicillium,
Coprinellus, Fusarium,
Rhizoctonia and Stemphylium
genera
candidum
A. niger, Aspergillus  
ochraceus, A. sydowii,
Aspergillus ustusa,
Botrytis cinerea, Eurotium
herbariorum, Gonytrichum
macrocladum, Penicillium
decumbensa, Penicillium
expansum, Penicillium
hirsutum, Penicillium
polonicuma, Penicillium
sp., Periconia britannicaa,
Rhizopus stolonifer,
S. chartarum and Ulocladium
botrytisa
  Efficacy  
  The extract of glycoalkaloids from  
nightshade plants demonstrated only
partial growth inhibition of Fusarium and
Rhizoctonia
  Highest antifungal activity shown  
by clove, lemon, bitter orange and
peppermint. The concentration of 5 ppm
was as effectivity as 5 ppm Ketoconazole – Efficacy compared to
(positive control) and the lowest was
shown by castor oil, cedar and olive
Thujopsene demonstrated fungicidal  
activity against only 5 of the 16 fungal
strains tested
Strengths  
– Relevant fungal species used  
(isolated from building surfaces) real-world application (percentage of
– Relevant fungal species used  
(isolated from indoor air and
surfaces)
commercially available fungicide centre with the antifungal agent)
as positive control
– Relevant fungal species used
(isolated from indoor air
environment)
– Fungicidal ability tested against
a large number of fungal
isolates
– Multiple time points assessed
Weaknesses   Ref.
– Limited transferability of results to   (35)
growth inhibition was calculated using
the agar micro-dilution method)
– Incubation time was describes as
4–5 days. The effect of time was not
assessed
– Efficacy was not compared to
commercial fungicide
– Limited transferability of results to   (36)
real-world application (MIC were
determined using potato dextrose
agar plate needle-inoculated in the
– Multiple time points were not
assessed
  – Limited transferability of results to   (37)
real-world application (disk diffusion
and agar well diffusion assay on malt
extract agar and fungicidal activity
reported as growth inhibition zones)
– Efficacy not compared to
commercially available fungicide
Whiley et al.: Indoor air and antifungal essential oils      69
Table 1 (continued)

Treatment   Fungal species   Efficacy   Strengths   Weaknesses   Ref.

Cedar leaf oil (Thuja plicata)  
Essential oil extracts from  
manuka (Leptospermum
scoparium) including eugenol,
thymol, cinnamaldehyde,
carvacrol, manuka oil, manuka
oil less triketones fraction and
triketones
Candida albicans and A. niger   C. albicans was readily killed by cedar leaf  – Moderate transferability of
oil. A. niger was inhibited but complete
eradication was not achieved
Penicillium corylophilum,   Eugenol, cinnamaldehyde, thymol and  
A. alternata and Cladosporium carvacrol completely inhibited the growth
herbarum of the three test fungi at a concentration
of 1% w/v
All the extracts, including eugenol,
cinnamaldehyde and thymol virtually
completely inhibited the growth of
P. corylophilum on unfinished gypsum
board at 3% w/v and significantly
reduced growth on the finished gypsum
boards
  – Unknown relevance of fungi to   (38)
results to real-world application environmental isolates (ATCC strains
(dried films of fungi were used)
exposed directly to cedar leaf oil) – Efficacy not compared to
– Multiple time points assessed commercially available fungicide
– Moderate transferability of   – Source of fungal isolates not reported   (39)
results to real-world application – Efficacy not compared to
(initial experiment – growth commercially available fungicide
inhibition was examined on malt
extract agar containing essential
oil compared to growth of the
control. Follow-up experiment –
the efficacy of eugenol,
thymol and cinnamaldehyde
was evaluated against P.
corylophilum on gypsum board.
70      Whiley et al.: Indoor air and antifungal essential oils

Finished (sealed and painted)

Thyme essential oil and thymol  
Aspergillus spp. A. versicolour,   Both thymol and thyme essential oil
A. niger, A. sulphureus, A.
flavus, P. chrysogenum,
P. brevicompactum, P.
griseofulvum, Penicillium
spp., Alternaria spp., A.
alternat, Ulocladium spp.,
Absidia spp., Mucor spp., C.
spp., C. sphaerospermum,
Trichoderma spp., Rhizopus
spp., C. globosum, and
S. chartarum
 
showed strong fungicidal activity
Vapor phase of the thyme essential oil
at concentration of 82 μg L−1 exhibited
fungi-static and/or fungicidal activity
during 60 days of exposure in glass
chambers. They were also demonstrated
to be sporicidal against all tested mould
species
and unfinished gypsum was
sprayed with the essential oil
treatment and then inoculated
with 107 spores/mL)
– Multiple time points assessed
– Relevant fungal species used   – Efficacy was not compared to   (40)
(isolated from building surfaces) commercially available fungicide
– Moderate transferability of – Multiple time points were not
results to real-world application assessed
[initial experiment – MIC and
MFC were determined by using
the serial broth dilution method.
Follow-up experiment – the
activity of the vapour phase of
essential oil of thyme was tested
by a modified micro-atmosphere
method]
Table 1 (continued)
Treatment  
Essential oils from lemon  
(C. limon) including C. paradishi,
C. sinensis, Citrus aurantifolia
and Citrus reticulate
Volatile organic compounds  
(VOCs) produced from
evaporating essential
oils indoors lavender
(L. angustifolis), eucalyptus
(E. globulus) and tea tree
(M. alternifolia)
Essential oil from pine tree  
(Pinus sylvestris L.)
DMSO, Dimethyl sulfoxide; ATCC, American type culture collection.
Fungal species  
A. niger  
Natural fungi contained within   A slight decrease in fungal concentrations   – Comparatively transferable to
air samples
A. niger, Penicillium  
funiculosum, P. chrysogenum, against all fungi tested, although the
T. viride, Ulocladium
oudemansii, Paecilomyces
variotii, Phoma glomerata, S.
chartarum and A. versicolor
Efficacy  
C. aurantifolia and C. reticulata exhibited     – Source of fungal isolates not reported   (41)
significant antifungal potency against
building fungi
was observed in the first 30–60 min but
levels quickly increased to pre-treatment
concentrations
Pine oil displayed fungicidal activity  
effectiveness depended on the fungal
species and the concentration of pine oil
Strengths  
 
real-world application [300 μL
of each essential oil was diluted
with 50 mL water for use in
incense evaporator with burning
candle in two rooms (21.6 m3
and 28.2 m3). Changes in total
airborne fungal concentration
were examined by air sample
collection using Burkard
sampler and enumerated on
malt extract agar plates an hour
prior to treatment]
– Relevant source of fungi used
– Multiple time points assessed
– Relevant fungal species used  
(isolated from indoor air
environment)
Weaknesses   Ref.
– Multiple time points not assessed
– Efficacy was not compared to
commercially available fungicides
– Limited transferability of results to
real-world application (MICs were
calculated at different concentrations
of essential oils on potato dextrose
agar)
– Efficacy not compared to   (23)
commercially available fungicide
– Limited transferability to real-world   (42)
application [MICs were determined
using oil diffusion on Czapek agar (for
fungi) and malt extract agar (for yeast
and yeast-like fungi)]
– Multiple time points not assessed
– Efficacy not compared to
commercially available fungicide
Whiley et al.: Indoor air and antifungal essential oils      71
72      Whiley et al.: Indoor air and antifungal essential oils
number of studies (laboratory and in situ) that have
demonstrated that clove oil has strong antifungal capa-
bilities. The most robust study demonstrated that clove
essential oil had fungicidal activity comparable to com-
mercial disinfectants and had the highest antifungal
efficacy compared with Siberian fir, common caraway,
peppermint, willow-leaved gum tree, lemon thyme and
bergamot orange against fungi isolated from indoor air
and surfaces (A. versicolor, A. niger, A. fumigatus, C.
sphaerospermum, C. cladosporioides, P.  chrysogenum,
P. aurantiogriseum, P. simplicissimum, U. chartarum and
P. digitatum) (34). Another study that was designed to
have moderate transferability of results, by Yingpra-
sert et  al. (31), demonstrated that particle board that
had been dipped in 0.63% clove oil completely inhib-
ited Aspergillus and Trichothecium for up to 5 weeks. By
increasing the concentration to 3%, it was found that
Aspergillus and Trichothecium were inhibited for up
to 9 weeks and the percentage of mass lost as a conse-
quence of Trametes and Gloeophylum was reduced by
5% (31).
These findings were supported by other studies that
used agar plates spiked with clove oil to demonstrate that
it had higher antifungal efficacy against A. niger and G.
candidum compared with black pepper, castor-oil plant,
cedar, eucalyptus and olive (36). Also, a study using the
agar spiked method found that clove oil was 100% effec-
tive at controlling R. chinensis, a common white rot fungi
found in wood surfaces (29).
Tea tree oil
Two studies from Table 1 identify tea tree oil as a poten-
tial antifungal agent. The most translatable study was Su
et al. (23), which evaluated tea tree oil by evaporating it
in a room and measuring the changes in fungal concen-
tration using an air sampler. This study is one of the few
investigations into applying essential oils as a fungicide in
a real-world building environment. It was found that the
concentrations of fungi in the air initially decreased as a
result of the VOCs from tea tree oil, but after 30–60 min,
the concentrations returned to normal background indoor
levels. However, one of the limitations of this study was
that it did not compare the efficacy of tea tree oil to that
of commercially available fungicides. Another study used
the disk diffusion assay method to test tea tree oil against
A. fumigatus and P. chrysogenum isolated from indoor air
samples and found that it had greater fungicidal activity
compared with some commercially available antifungal
agents (15).
Oregano
Oregano was identified as an effective antifungal agent
in a study by Zabka et  al. (33) using the agar microdilu-
tion method. It was demonstrated to have high inhibition
levels against all fungi tested (A. alternata, S
­ . chartarum,
C. cladosporioides and A.  niger) and was more effective
compared to other essential oils, including English lav-
ender, pennyroyal, corn mint, sage, bitter orange, berga-
mot orange, common juniper, basil, common sage, tansy
and ginger. However, this study used stock culture fungi,
which makes it difficult to translate these results com-
pared to studies using environmentally isolated fungi,
and did not compare efficacy to that of commercially
available fungicides. These findings were supported by
another study that also used the microdilution methods
to demonstrate that oregano displayed antifungal prop-
erties against A. fumigatus, A. nidulans, A. versicolor and
a Penicillium species isolated from the frescoes within a
monastery in Serbia. However, it was not as effective com-
pared to the biocide benzalkonium chloride, a quaternary
ammonium compound (32).
Thyme
Thyme was also identified as an effective antifungal agent
in a study by Zabka et  al. (33), with greater fungal inhi-
bition potential compared to essential oils from English
lavender, pennyroyal, corn mint, sage, bitter orange,
bergamot orange, common juniper, basil, common sage,
tansy and ginger. However, this study used stock culture
fungi, which makes it difficult to translate these results
compared to studies using environmentally isolated
fungi, and did not compare efficacy to that of commer-
cially available fungicides. It was also demonstrated to
have fungicidal and sporicidal activity in both the liquid
and vapour phases against fungal isolates collected from
the walls of damp buildings in Slovakia (40). However, in
other more robust studies, it was shown to be not as effec-
tive compared to clove oil, fir oil or marjoram (28, 34).
Citrus
Verma et al. (41) demonstrated that the volatile essential
oils from lime (C. aurantifolia) and mandarin (C. reticulate)
exhibited significant antifungal potency against build-
ing fungi A. niger. This was followed up by another study
using a modified disk diffusion assay method (the essen-
tial oil was needle-inoculated in the centre of the agar

plate), which showed that 5 ppm concentration of lemon
essential oil was as effective as 5 ppm ketoconazole (a syn-
thetic antifungal drug) against A. niger and G. candidum
isolated from the surfaces and indoor air of buildings. The
study also demonstrated essential oil from lemon to have
greater antifungal potential compared to castor oil, cedar
and olive (36).
Other essential oils demonstrating
­fungicidal potential
Other essential oils that demonstrated potential antifun-
gal activity in at least one study (Table 1) include heart-
wood, marjoram, cinnamon, lemon basil, caraway, bay
tree, fir, peppermint, pine, cedar leaf and essential oil
extracts from manuka (eugenol, cinnamaldehyde, thymol
and carvacrol).
Essential oils demonstrating only moderate
or low fungicidal activity
The essential oils identified in Table 1 which demonstrate
only moderate or limited antifungal activity include euca-
lyptus leaves, crêpe ginger, ginkgo leaves, myrrh, English
lavender, pennyroyal, corn mint, sage, bitter orange, ber-
gamot orange, common juniper, common basil, night-
shade, castor-oil-plant, olive, willow-leaved gumtree and
thujopsene (a compound found in the essential oil of a
variety of conifers).
Mechanism of antifungal activity
Understanding the mechanism(s) of action of different
antimicrobial agents is important to characterise effi-
cacy as one agent may not inhibit all microorganisms.
It is important to acknowledge the principal differences
between bacteria and fungi. The structures of fungi and
bacteria differ in significant ways, for example most fungi
are diploid in nature and have longer generation time
compared with bacteria (46). This means that antibac-
terial and antifungal agents target structures and func-
tions most relevant to the organisms to be inhibited. For
example, many antibacterial agents inhibit steps impor-
tant for the formation of peptidoglycan (47), the essential
component of the bacterial cell wall. In contrast, most
antifungal compounds target either the formation or the
function of ergosterol (48, 49) an important component
of the fungal cell membrane. This membrane interaction
Whiley et al.: Indoor air and antifungal essential oils      73
weakens the structure, increasing permeability, which is
responsible for the leakage of solutes across the mem-
brane and causes cell lysis. For example, Shao et al. (50)
described this mechanism of action when applying tea
tree oil on B. cinerea (an important fungus in viticulture
and food spoilage). Tea tree oil was found to inhibit the
growth of the fungus and germination of spores was sup-
pressed. The cell wall structure was reported to have lost
its ultrastructure and showed thickening and rupturing.
The authors concluded that the cell wall integrity was
destroyed, increasing the membrane permeability.
Overall, there is currently limited knowledge regard-
ing the antimicrobial mechanisms of essential oils, par-
ticularly with regards to antifungal activity (36, 49). A
few authors have mentioned the antimicrobial activity of
essential oils; however, the mechanism of action has not
been studied in great detail (49, 51). Chemical analysis
of essential oils show that the major active components
are phenols, terpenes, aldehydes and ketones (52), and
it is generally believed that essential oils principally act
against cell cytoplasmic membranes of microorganisms.
Hydrophobicity is an important characteristic of essential
oils and their components (51), which may enable them to
accumulate in cell membranes, disturbing the structures
and causing an increase of permeability.
One study by Pinto et al. (49) demonstrated that clove
oil and eugenol (the main component of many essential
oils) was found to be fungicidal as a result of extensive
lesion of the fungal cell membrane. In addition, clove oil
and eugenol reduced the quantity of ergosterol, a specific
fungal cell membrane component. This resulted in inhibi-
tion of germ tube formation of C. albicans (49). Similarly,
it has been suggested that the antifungal action of tea tree
oil is as a result of its capability to change or damage the
function of fungal membranes (50, 53).
A great deal remains to be learned about the mecha-
nisms of action of essential oils against fungal species.
Although some progress has been made with clinical
investigations, a greater understanding of these mecha-
nisms is clearly lacking for other environmental organ-
isms. Studies of the mechanisms of action relevant to
fungal species in indoor air would allow more efficient
and effective use of these agents.
Potential health effects – is ‘natural’ safer?
The increasing interest in ‘natural’ products for controlling
microorganisms in indoor environments is due in part to
the perception of benefit (i.e. inhibition of fungal growth)
without the need for using potentially ‘harmful synthetic
74      Whiley et al.: Indoor air and antifungal essential oils
chemicals’. However, this assumption that ‘natural’ prod-
ucts are not harmful to human health is flawed.
Currently, there are limited studies investigating the
potential adverse health consequences of repeated expo-
sure to essential oils. The oils themselves are complex
mixtures, which may contain naturally occurring contact
sensitisers. In fact, some evidence suggests that they are
potential skin allergens or sensitising agents (23, 54, 55).
An ideal antifungal agent would not generate toxic fumes
during application and is non-irritating if accidentally
exposed to skin. Skin irritation and skin sensitisation are
different responses; skin irritation occurs on the first expo-
sure to the agent; the inflammatory reaction is typically
rapid and the severity will depend on the concentration
of the irritant present, compared with skin sensitisation,
which is a complex allergic immunological response, with
the reaction typically occurring after repeated exposure to
the chemical and is usually irreversible (i.e. once sensi-
tised, always react). Schaller and Korting (55) described a
case report of allergic contact dermatitis due to repeated
exposure to essential oil use in aromatherapy (applied
topically or released as aerosols). There have also been
several studies that have demonstrated exposure to
essential oils exacerbated respiratory problems includ-
ing asthma, decreased pulmonary function and increased
chest tightness (56, 57).
Thus, the perception that ‘natural’ is safer may not
necessarily be appropriate when considering essential oils
for fungicidal treatment, and care must be taken with their
repeated application in the indoor environment. Essen-
tial oils should be considered in the same way that use of
chemical fungicides would be, based on risk assessment.
Conclusion and recommendations
Fungal contamination of indoor buildings and indoor air
quality is a health issue of increasing concern. There is a
need for greater guidance regarding appropriate antifun-
gal agents to treat fungal contamination of buildings, as
well as a drive from consumers and other groups to con-
sider the potential of essential oils as a ‘natural’ alterna-
tive to commercially available fungicides. This review
identifies the studies assessing the fungicidal potential of
essential oils against fungi relevant to indoor air quality.
The biggest challenge with comparing the fungicidal effi-
cacy of essential oils from different studies is the lack of
a standard method and reporting language. Addition-
ally, the efficacy of essential oils is also dependant on the
fungal species being challenged which makes it difficult
to compare studies using different fungal isolates.
However, despite these challenges, clove oil was iden-
tified as the best-performing essential oil within the more
robust studies. Additionally, there appears to be some evi-
dence to support the essential oils tea tree oil, oregano,
thyme and lemon as potential antifungal agents with rel-
evance to indoor air quality. Heartwood, marjoram, cinna-
mon, lemon basil, caraway, bay tree, fir, peppermint, pine,
cedar leaf and manuka were also identified in at least one
study as having antifungal potential; however, there is a
need for more robust studies to examine these further.
Future studies should focus on comparing the effi-
cacy of these essential oils against a large number of
fungal isolates from indoor environments. Studies will
then need to focus on translating these results with in
situ studies investigating the effectiveness in actual build-
ings and assessing the potential for long-term antifungal
persistence. The studies identified in this review, which
were either moderately or comparatively translational
(23, 28, 31, 38–40), can inform the design of these studies.
However, they should additionally compare the efficacy
of essential oils to commercially available fungicides and
examine the effect of time on fungicidal activity. Further-
more, when considering the application of these essential
oils in building environments, the effect of different con-
centrations, mechanisms of application and the potential
human side effects must also be examined.
Author Statement
Research funding: Authors state no funding involved.
Conflict of interest: Authors have no conflicts of interest to
declare. Informed consent: Informed consent is not appli-
cable. Ethical approval: The conducted research is not
related to either human or animal use.
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