Shared Makeup Cosmetics as a Route of Demodex folliculorum Infections

Abstract

Purpose
The aim of the study was to examine Demodex survival in makeup cosmetics, i.e., powder cream, mascara, and lipstick, and to determine whether cosmetics shared with others can be a source of D. folliculorum infection.

Methods
Live D. folliculorum adults were placed in cosmetic samples and their motility was observed under a microscope. The mites were fully or partially immersed in the powder cream and lipstick, and only partially immersed in the mascara. Partial immersion means that only the opisthosoma was covered by the cosmetic, whereas the gnathosoma and podosoma had no contact with the cosmetic. Cessation of motility was regarded as a sign of death.

Results
In the control (mites placed on a microscope slide with no cosmetics), the survival time was 41.2 h. D. folliculorum that were immersed fully or partially in the lipstick substrate were viable for 38.5 h and 148 h, respectively. The survival time of the mites at full and partial immersion in the powder cream was 0.78 h and 2.16 h, respectively. The average survival time in the mascara was 21 h.

Conclusions
Makeup cosmetics used by different individuals at short intervals (from several hours to several days) can be a source of transmission of Demodex sp. mites.

Full text

Vol.:(0123456789)
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Acta Parasitologica
https://doi.org/10.1007/s11686-020-00332-w
ORIGINAL PAPER
Shared Makeup Cosmetics asaRoute ofDemodex folliculorum
Infections
AleksandraSędzikowska1 · KatarzynaBartosik2 · RenataPrzydatek‑Tyrajska2,3 · MonikaDybicz1
Received: 14 October 2020 / Accepted: 19 December 2020
© The Author(s) 2021
Abstract
Purpose The aim of the study was to examine Demodex survival in makeup cosmetics, i.e., powder cream, mascara, and
lipstick, and to determine whether cosmetics shared with others can be a source of D. folliculorum infection.
Methods Live D. folliculorum adults were placed in cosmetic samples and their motility was observed under a microscope.
The mites were fully or partially immersed in the powder cream and lipstick, and only partially immersed in the mascara.
Partial immersion means that only the opisthosoma was covered by the cosmetic, whereas the gnathosoma and podosoma
had no contact with the cosmetic. Cessation of motility was regarded as a sign of death.
Results In the control (mites placed on a microscope slide with no cosmetics), the survival time was 41.2h. D. folliculorum
that were immersed fully or partially in the lipstick substrate were viable for 38.5h and 148h, respectively. The survival
time of the mites at full and partial immersion in the powder cream was 0.78h and 2.16h, respectively. The average survival
time in the mascara was 21h.
Conclusions Makeup cosmetics used by different individuals at short intervals (from several hours to several days) can be
a source of transmission of Demodex sp. mites.
Keywords Demodicosis· Demodex folliculorum· Demodex transmission· Transmission via cosmetics· Blepharitis
Supplementary Information The online version contains
supplementary material available at https ://doi.org/10.1007/s1168
6-020-00332 -w.
* Aleksandra Sędzikowska
aleksandra.sedzikowska@wum.edu.pl
1 Department ofGeneral Biology andParasitology, Medical
University ofWarsaw, Chałubińskiego 5, 02-004Warszawa,
Poland
2 Chair andDepartment ofBiology andParasitology, Medical
University ofLublin, Radziwiłłowska 11, 20-080Lublin,
Poland
3 Reno-Med Non-Public Health Care, Podkowy 87,
04-937Warszawa, Poland
Introduction
Demodex mites are permanent residents of pilosebacious
units in humans and other mammalian species. Two spe-
cies, i.e., Demodex folliculorum and Demodex brevis, have
been found to infest humans. The predilection sites mainly
include the face, scalp, and chest, but the mites can also
colonise other parts of the body [1, 2]. D. folliculorum
is most often present in hair follicles, whereas D. bre-
vis attacks sebaceous glands. The infestation can be both
symptomatic and asymptomatic. Such ocular symptoms as
itching, redness of eyelids, and lacrimation are reported
most frequently [3]. Ocular demodicosis is also considered
as a risk factor of recurrence of pterygium [4]. In some
patients, the presence of these mites may be associated
with blepharitis, rosacea, chalazion, perioral dermatitis,
or idiopathic follicular mucinosis of the head and neck
[5–10]. Cases of local dermatological lesions caused by
increasing Demodex populations have been reported as
well [11].
In the case of mites that are permanent human parasites,
such as Demodex spp. or Sarcoptes scabiei, it is believed
that the invasion requires human skin contact with live
mites directly or via an indirect route.]. 241 sequences
from the mitochondrial genome of D. folliculorum were
analyzed and the haplotypes were much more likely to be
shared within families than between unrelated individuals.
Molecular analyses indicate that frequent close physical
contact leads to mite transmission [12]. Direct contact
or eggs present in dust as well as contact with infected
towels, blankets, or sponges are the possible routes of

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Key Points
This study investigates the relationship between sharing
cosmetics and Demodex folliculorum infection.
The results show that mites can survive from few hours
to few days in makeup cosmetics depending on its type
and composition.
Clients in drugstores using makeup cosmetic testers are
at risk of Demodex folliculorum transmission via this
route.
than in juvenile stages and the movement of their legs is
more clearly visible. All adult forms were observed under a
microscope every 5min and their continuous motility was
recorded. The analyses of mite viability in the cosmetics
were carried out only on specimens that moved all the legs
and gnathosoma. Mites showing minimal movement (e.g.,
only the gnathosoma or one pair of legs) were not qualified
for further tests. Samples marked as positive were used in
the study on the day of sampling.
The survival of Demodex mites was examined in basic
makeup cosmetics, i.e., mascara, lipstick, and powder cream.
Each cosmetic was placed on a microscope slide. Next, the
mites were gently transferred directly onto the tested cos-
metic and no coverslip was used. A single mite was placed
on each microscopic slide. Demodex mites transferred
to a slide with the cosmetic did not change the place but
only moved their legs and gnathosoma and stayed on the
slide. Colourless lipstick was used to facilitate the observa-
tion of the mites. Demodex could be on the surface of the
cosmetic or deeper that is why they were immersed either
fully or partially in the powder cream and lipstick. Partial
immersion means that only the opisthosoma (fragment C
marked in Fig.1) was covered by the cosmetic, whereas the
gnathosoma and podosoma (fragments A and B marked in
Fig.1) had no contact with the cosmetic. Specimens were
transferred onto the mascara without full immersion, as its
Fig. 1 Morphological elements of Demodex folliculorum: A – gna-
thosoma, B – podosoma, C – opisthosoma (original magnification
200x)
Demodex spp. infection described in the literature [13, 14].
The use of facial creams or eyeliners has been considered
as a potential route of Demodex spp. infections; however,
no studies have addressed the issue of the length of sur-
vival of these mites in cosmetics.
Cosmetics are shared not only in households. Before
purchase, cosmetic testers available in drugstores can be
used or makeup can be done as a special offer by a beauti-
cian, who uses the same cosmetics in many customers.
Whether such consumer behaviour can become a potential
route of Demodex spp. infection is unknown. Therefore,
the aim of the study was to determine the length of sur-
vival of Demodex mites in commonly used makeup cos-
metics, i.e., powder cream (fluid foundation), mascara, and
lipstick, and to find out whether shared cosmetics could be
a source of D. folliculorum infection.
Materials andMethods
Research Procedures
Live Demodex mites collected from patients reported for
consultation with an aesthetic medicine doctor in Warsaw,
Poland. The investigation procedures involving the volunteer
patients in the study were approved by the local Bioethics
Committee at the Medical University of Lublin (approval no.
KE-0254/122/2018). Individual patients’ consent to partici-
pate in the study was obtained as well. The mites were col-
lected from the patients with a method of lash sampling with
sterile tweezers. Eyelashes from the left and right eye were
sampled for the study. The eyelashes were placed directly on
the microscope slide. A Zeiss Primo Star light microscope
(magnification from 40 × to 400 × ) was used to detect the
presence of Demodex spp. in the collected material. The
presence of Demodex spp. adults, juvenile forms, or eggs
on the slide indicated that the sample was positive. Further
analyses were performed only on positive slides with the
presence of adults (Fig.1), as their legs are better developed

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dark colour would hamper observations of these small mites.
Demodex mites placed on the microscope slide with no cos-
metics were the control sample. No coverslips were placed
on the slides with the mites. All samples were transferred
to a humid chamber to prevent desiccation. The slides were
stored at room temperature of 22°C and 60% RH to mimic
drugstore and home storage conditions.
The Demodex spp. specimens were observed under a light
microscope at various intervals. Their viability was assessed
according to the following criteria:
• (+ + +) high viability (the mite moved the legs and gna-
thosoma; sometimes, the entire body moved)
• (+ +) moderate viability (the mite moved the legs and
gnathosoma but at a definitely slower pace)
• (+) low viability (the mite exhibited minimal movements
of the claws only or gently moved the gnathosoma)
• (−) no motility (the mite did not move any part of the
body during the observation).
Mites assigned to categories (+), (+ +), and (+ + +) were
considered alive. Each specimen that showed no motility
was observed three more times at a several-minute inter-
val. When the subsequent observations revealed no signs of
life, the specimen was regarded dead and the first time point
of cessation of movement was recorded. The frequency of
observations depended on the motility of the mites. In the
case of the very high viability (+ + +), the observations were
conducted every 2–4h. When the viability declined with
time (+), the observations were conducted more frequently
(every 5–10min) to record the time point of cessation of
movement. Demodex mites with low viability (+) were
observed at high magnification (400 ×) to notice even the
slightest movements indicating that the specimen was still
alive. Sometimes, only a claw or a part of the gnathosoma
was found to move. In the case of an immotile specimen, the
exact time of cessation of movement was recorded, and the
mite was checked three times to monitor any signs of life.
When there was no movement in each subsequent observa-
tion, the specimen was assumed to have died at the first time
point recorded.
Commonly available facial cosmetics, i.e., powder cream,
mascara, and lipstick, were selected for the study. The chem-
ical composition of the cosmetics including the first six
ingredients with the highest concentration was as follows:
Powder cream: dimethicone, water, dipropylene glycol,
alcohol denat., nylon-12, butylene glycol.
Lipstick: cera microcristallina, octyldodecanol, hydrogen-
ated polydecene, Ricinus communis seed oil, cetyl palmitate,
ethylhexyl methoxycinnamate,
Mascara: water, Copernicia cerifera cera, cera alba, glyc-
eryl stearate, Euphorbia cerifera cera, stearic acid.
Statistical Analysis
The following tests were used to check the difference in the
survival of the mites in the different cosmetics:
– Student’s t test–comparison of the survival of the mites
in two groups (normality of distribution and uniformity
of variance),
– Cochran-Cox test–comparison of the survival of the
mites in two groups (normality of distribution but no
homogeneity of variance),
– Mann–Whitney U test–comparison of the survival of the
mites in two groups (no normality of distribution),
– Kruskal–Wallis ANOVA test–comparison of the survival
of the mites in six groups (no normality of distribution
and no homogeneity of variance). The Kruskal–Wallis
multiple comparison (post-hoc) test was used to analyse
the significance of the differences in the survival between
the substrates.
A p value of < 0.05 was considered statistically signifi-
cant. Statistical calculations were carried out using the STA-
TISTICA 10 PL statistical package.
Results
From 10 to 18 D. folliculorum adults, i.e., the species present
in the examined patients, were subjected to observations in
each cosmetic substrate. In total, the survival of the mites
was examined in 77 samples (28 samples of the lipstick, 25
samples of the powder cream, 14 samples of the mascara,
and 10 control samples). The invitro survival time of the
Demodex mites in the selected cosmetics and in the control
sample is presented in Table1.
The survival time was the longest in the case of D. fol-
liculorum immersed in the lipstick- 69h and 260h for the
fully and partially immersed mites, respectively (Fig.2). In
the mascara, the mites were able to stay alive for as long as
56h. In the powder cream samples, the overall survival time
of D. folliculorum was the shortest, i.e., maximum 2.3 and
4.5h for the fully and partially immersed mites (Table1).
Discussion
Human sebum, which consists mainly of triglycerides and
fatty acids (57.5%) as well as wax esters (26%), squalene
(12%), and cholesterol (4.5%), is an optimal living environ-
ment for D. folliculorum and D. brevis [15, 16]. These com-
pounds protect the mites against water loss and are a source
of food. Hormones increasing sebum production may also

Acta Parasitologica
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affect the course of demodicosis and increase the possibility
of infection. Demodicosis and seborrheic dermatitis have
been associated with polycystic ovary syndrome (PCOS),
probably due to the increased discharge of sebum promoted
by androgens. D. folliculorum has been found to be more
predominant in women with PCOS [17]. An important epi-
demiological task is to determine whether and how long
these permanent human parasites can survive without a host.
Our previous investigations demonstrated that Demodex spp.
immersed in saline survived on average for 82h [18]. In the
present study, the mites in the control group survived for
41h, which was shorter than in physiological saline. The
shortest viability time was recorded in the powder cream
substrate. The survival time in the mascara was longer, and
the lipstick substrate ensured the longest viability of the
mites. The analysis of the Demodex survival in the cosmetics
was focused on the first six chemical components that may
potentially affect the viability of these mites. Demodicidal
activity has been shown for many compounds, i.e., 100% tea
tree oil (TTO), 100% caraway oil, 99% ethanol, 10% povi-
done-iodine, 4% pilocarpine, and 4% terpinen-4-ol (T4O),
but only some of them can be used as cosmetic ingredients
due to their intrinsic toxicity and the risk of irritation of eyes
and skin [19–22].
The powder used in the study contained the following
ingredients from the top of the list: dimethicone, water,
dipropylene glycol, alcohol denat., nylon-12, and butylene
glycol. Dimethicone and denatured alcohol seem to be the
most important components influencing Demodex survival.
Dimethicone has a protective effect, as it protects the skin
against external factors. It forms a film on the skin produc-
ing a feeling of smoothness, softens the skin, and nourishes
the hair [23]. The compound is used not only as a cosmetic
ingredient but also as an agent against head lice Pediculus
humanus capitis [24]. It enters the respiratory system of lice
and causes their death within 0.5h [25]. Other authors sug-
gest that the death of lice does not occur via hypoxia but via
inhibition of water excretion [26]. The average survival time
of Demodex mites that were fully immersed in the powder
cream was comparable to that of dimethicone-treated head
lice (0.78h). Mites that were partially immersed in the pow-
der cream survived over twice as long (2.16h). This may
indicate that dimethicone had the most significant effect on
the survival of Demodex in this substrate.
Another component, i.e., dipropylene glycol (DPG), is
used e.g. as a solvent in cosmetics and personal care prod-
ucts [27]. There are no data on the effect of this substance on
mites. Alcohol denat. is a general term used by the cosmetics
industry to refer to denatured alcohol, which is added to a
wide range of products [28]. It acts as an effective preserva-
tive, prevents spoilage of cosmetics, and serves as an anti-
bacterial agent [29]. In 75% and 100% ethyl alcohol, Demo-
dex can survive for approx. 150 and 4min, respectively [20].
The concentration of the alcohol in the powder cream is
definitely lower; however, its presence may be associated
with the shorter survival time of the mites in this substrate.
Since it is usually applied to the face with fingers, powder
cream can be a source of Demodex infections when used
by several persons within a short time (tens of minutes). It
seems that the dimethicone ingredient is crucial for Demo-
dex survival in this cosmetic.
The Demodex mites were immersed in the mascara par-
tially, as its dark colour would have prevented observation
at full immersion. The small size of these mites impeded
Table 1 Overall survival of
Demodex folliculorum in the
cosmetics and control group
(descriptive statistics)
SD standard deviation, n number of samples
p < 0.05, (1) Student’s t test, (2) Cochran-Cox test, (3) Mann–Whitney U test
Substrate nSurvival time (h) p
Mean SD Median Min Max
Control 10 41.2 22.2 42.0 9.0 84.0 0.7269(1)
Lipstick–full immersion 18 38.5 17.7 36.5 7.0 69.0
Lipstick–partial immersion 10 148.1 63.1 147.0 67.0 260.0
Powder cream–full immersion 15 0.79 0.64 0.52 0.25 2.30 0.0000(3)
Powder cream–partial immersion 10 2.17 1.11 1.86 0.88 4.53 0.0004(2)
Mascara–partial immersion 14 21.4 14.8 19.0 2.5 56.0 0.0154(1)
Fig. 2 Adult Demodex folliculorum fully immersed in colourless lip-
stick (original magnification 200x)

Acta Parasitologica
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observations of the movement of legs, which was the basic
survival criterion. The partially immersed specimens were
found to survive in the mascara for 21h on average.
The first main components of the mascara are water,
Copernicia cerifera cera, cera alba, glyceryl stearate,
Euphorbia cerifera cera, and stearic acid. Water provides
Demodex mites with the required humidity; hence, this sub-
stance does not seem to have a negative effect on the sur-
vival of these parasites. Another ingredient, i.e., Copernicia
cerifera cera, is wax extracted from leaves and buds of the
Brazilian palm Copernicia prunifera. This substance is con-
sidered hypoallergenic, although allergic contact dermatitis
has been reported in the literature [30]. In turn, the cera
alba beeswax forms an occlusive layer on the surface of the
skin or mites, preventing excessive evaporation of water. In
humans, cera alba may be a causative agent of allergic reac-
tions [31]. Euphorbia cerifera cera is a wax component as
well; yet, the effect of waxes on Demodex mites is unknown
at present [32].
Glyceryl stearate is used in cosmetic preparations as an
emollient, emulsifier, and stabiliser. This component was
found to be slightly toxic in acute oral toxicity studies in rats
[33]. However, as in the case of waxes, its effect on Demodex
has not been described.
Demodex mites immersed in the lipstick substrate had
the longest survival time (fully immersed: 38.5h, partially
immersed: 148.1h). The main components of this product
include various types of emollients preventing water evapora-
tion. Cera microcristallina (crystalline mineral wax), i.e. the
so-called greasy emollient, is the first ingredient on the list. It
is used in skin and hair care formulations to form an occlu-
sive layer (film) on their surface. Thus, it prevents excessive
evaporation of water. Similar effects are provided by the other
ingredients, i.e., octyldodecanol, hydrogenated polydecene,
Ricinus communis seed oil, and cetyl palmitate [34–37]. In
turn, ethylhexyl methoxycinnamate is one of the most com-
monly used UVB filters in sunscreen products [38]. There are
no data about its acaricidal activity.
Most compounds contained in the lipstick have lubricating
properties. Given the long survival of the Demodex mites in
this cosmetic, it can be concluded that the parasites find such
an environment favourable as they feed on lipids [39]. Trans-
mission via shared lipstick may take place mainly in the case
of perioral demodicosis.
The present study was conducted to determine whether
widely available cosmetics, e.g., testers in drugstores or cos-
metics used by makeup artists, could be a potential source of
Demodex infection. The study was focused on facial cosmetics,
as these parasites most frequently colonise the face. The high
prevalence of Demodex spp. in humans indicates widespread
occurrence of these mites, whereas the routes of their spread
have not been fully elucidated.
Our results indicate that mites can be spread in shared cos-
metics. Shared facial powder is associated with the likelihood
of transferring Demodex spp. when the cosmetic is used by
several persons within a short time (average survival time:
47min). Shared lipsticks may promote Demodex spp. infec-
tion (average survival time: 38.5h), although the number of
mites on the lips is inconsiderable. The analysed mites die
faster in powder, but there is a greater risk of transmission
thereof from the face onto the cosmetic with user’s fingers.
A high risk of transmission of Demodex spp. is associated
with using the same mascara, in which the mite was found to
survive up to 56h.
The survival of Demodex spp. in cosmetics is undoubtedly
influenced by their chemical composition as well as the mode
of use. Since testers in drugstores can be opened and applied
several times on the face with fingers instead of disposable
spatulas, there is a risk that mites present on the face will be
transferred to another person using the same tester. Testing
mascara poses a high risk, as Demodex mites are typically
present on eyelashes. The next person using the same brush
can be infected in this way.
Based on information provided by drugstore staff and our
observations, we estimate that from a dozen to even several
tens of persons per day may use makeup testers. Some of them
test such cosmetics as powder on the hand, but a large group
uses cosmetic testers for partial or full makeup. Similarly, cus-
tomers check mascara testers on their eyelashes. Such obser-
vations of consumer behaviour have supported our thesis that
investigation of the survival of Demodex mites in cosmetics is
an epidemiologically important issue.
Conclusions
Facial cosmetics shared at a short interval may contribute to
Demodex transfer between the users. Therefore, cosmetics
available to many customers should be tested with the use of
disposable spatulas and makeup cosmetics should only serve
for personal use. Furthermore, addition of not only bacteri-
cidal but also demodicidal compounds to makeup cosmetic
formulas should be considered.
Funding This research received no external funding.
Compliance with ethical standards
Conflict of interest The authors declare no conflict of interest.

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