International Biodeterioration & Biodegradation 117 (2017) 115e122

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International Biodeterioration & Biodegradation

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Indoor fungal contamination of traditional public baths (Hammams)

Leyla Benammar a, Taha Menasria a, b, *, Amel Chergui a, Soumia Benfiala a,
Ammar Ayachi c
a
Department of Microbiology and Biochemistry, Faculty of Natural and Life Sciences, University of Batna 2, Batna 05078, Algeria
b
Department of Applied Biology, Faculty of Exact Sciences and Natural and Life Sciences, University of Tebessa, Tebessa 12002, Algeria
c
Veterinary and Agricultural Sciences Institute, Department of Veterinary Sciences, University of Batna 1, Batna 05000, Algeria

a r t i c l e i n f o a b s t r a c t

Article history: This study was carried out to provide an overview of the fungal load in Algerian traditional baths
Received 17 September 2016 (Hammams) as well as to isolate and identify the main pathogenic fungi. Over a period of four months,
Accepted 7 December 2016 ten baths were examined and screened for fungal contamination from several parts of the hot steamy
rooms (floor, wall, door, air and marble massage platform). In total, 7157 fungi isolates were recovered
from the surveyed Hammams and the most abundant molds were Penicillium spp. (45.12%) followed by
Keywords:
Aspergillus spp. (28.80%). In addition, molds flora in traditional baths was characterized by a large
Indoor environment
number of hydrophilic species like Cladosporium, Fusarium, Rhizopus, Mucor and Alternaria. Eight
Public baths
Hammam
candida-like appeared frequently (C. albicans, C. glabrata, C. tropicalis, C. lipolytica, Geotrichum sp., Tri-
Fungal flora chosporon sp., Rhodotorula sp. and Cryptococcus sp.) of which C. albicans was the common isolated yeast
Candida albicans (35.14%). The results indicate a significant difference (p ¼ 0.007) in species richness between molds and
Dermatophyte yeasts and their distribution varied significantly among sampled positions in baths. ANCOVA revealed a
significant increase in fungal loads related to the average number of customers and mean opening year of
the Hammams, in contrast with locality (favored or popular district). This study indicates that Hammams
present a potential source of pathogenic fungi which may impose a real threat on public health.
© 2016 Elsevier Ltd. All rights reserved.

1. Introduction
Traditional baths, known as Hammams or Moorish baths are a
widespread culture in Maghreb countries including Algeria, Middle
East (mainly Syria) and Turkey (Bastuji-Garin et al., 2002; Ouaffak
et al., 2003; Goksugur et al., 2006; Benouis et al., 2008; Cherif-
Seffadj, 2009; Boggs, 2010). They are highly frequented public
places not only for personal hygiene but because Hammams were
considered for a long time as a cultural heritage originally attached
to religious and familiar rituals. Formerly, traditional bath was
adapted to the precepts of Islam, which calls for meticulous hygiene
and regular ablutions for prayer, performed as “Ghusl” (full body
cleansing) or “Wudu” (washing some parts of the body before each
prayers, five times per day). In most cases, Hammam is visited
frequently by women in order to recover from childbirth or illness,
as well it is considered as a real beauty corners related to different
* Corresponding author. Departement of Applied Biology, Faculty of Exact Sci-
ences and Natural and Life Sciences, University of Tebessa, Tebessa 12002, Algeria.
E-mail address: tahamenasria@hotmail.com (T. Menasria).
provided cares such as; hair removal, dyeing, application of henna,
scrubbing, massage and body care. In Islamic countries, women and
men alternatively use Hammams and it is strictly forbidden for the
simultaneous use, according to Islamic rulings.
Traditional baths are in general structured as enclosed place
consisting of a series of rooms; Bit Skhouna (hot steamy room), Bit
al Wasta (warm room), Bit al Barda (cold room) and al Bit El Q'ad
(the rest room) where people get wiping and dressing (Ouaffak
et al., 2003; Cherif-Seffadj, 2009). Nowadays, with the emergence
of private or individual baths and the progress of therapeutic vir-
tues of balneotherapy, usefulness of Hammams extends to be much
more associated with relaxation, well-being and even to treat or
prevent several dermatological and rheumatological illness
(Hannuksela and Ellahham, 2001; Matz et al., 2003; Verhagen et al.,
2003; Bender et al., 2005).
Fungal pathogens are recognized among leading agents that
causes human skin infections, accounting more than 25% of
affected population worldwide (Havlickova et al., 2008). Up to date,
over 400 pathogenic fungi species have been reported and some of
these are widely distributed in damp location such as swimming
pools, saunas, baths including Hammams, which were responsible

http://dx.doi.org/10.1016/j.ibiod.2016.12.004
0964-8305/© 2016 Elsevier Ltd. All rights reserved.
116 L. Benammar et al. / International Biodeterioration & Biodegradation 117 (2017) 115e122
for several infections (Chabasse et al., 2009; Matos et al., 2002;
Brandi et al., 2007). Hammams are highly suspected in mycosis
transmission, considering the high level of temperature and hu-
midity for long periods promoting fungal development (Havlickova
et al., 2008; Moriyama et al., 1992; Hamada and Abe, 2010). It was
reported that cutaneous infections specially induced by dermato-
phytes were mostly contracted in such places particularly Tinea
pedis, Tinea capitis and Onychomycosis (Kamihama et al., 1997;
Gudnadottir et al., 1999). In most cases superficial fungal in-
fections are contagious and may tend to recurrence (Kawachi et al.,
2010).
A part from, temperature and humidity which contributing to
fungi survival, poor hygiene and uncontrolled people attendance
with fungal skin infections, adnexa, and mucosae are certainly
remaining among the major sources of molds and yeast dissemi-
nation in traditional public baths. Many scientific reports deal with
the indoor mycobiota of various public and private buildings
(Hunter et al., 1988; Gilleberg et al., 1998; Picco and Rodol, 2000;
Gniadek et al., 2005; Nunes et al., 2013). However and despite the
increased importance of fungal contamination studies in traditional
hot steamy baths as part of preventive medicine against mycosis, no
researches are available on the incidence of indoor fungal diversity
and contaminants in Hammams in Algeria and North Africa.
Therefore, a quantitative and qualitative study was conducted to
determine and better understand the current distribution of indoor
fungal flora of traditional baths in a medium sized city with
approximately 320,000 inhabitants (Batna northeastern Algeria).
2. Materials and methods
2.1. Sample collection
Samples were collected from ten different Hammams in Batna
city (Northeast Algeria) over a period of four months (from
February up to May 2015). Hammams were selected randomly
including eight used alternatively by men and women and two
others used only by women. For each Hammam, two sets of sam-
ples were carried out before cleaning time on five different places
including floor, wall, door, air, and marble massage platform of
caladarium (hot steamy room). The first set was intended for the
enumeration of total fungal flora, while the second used for the
isolation.
For surface sampling, 10 cm2 selected sections was swabbed and
swabs expected for enumeration were placed in their tubes with
sterile saline, whereas those used for isolation in dry tubes
(Hamada and Abe, 2010). The air sampling has been carried out
using settle plate method (Napoli et al., 2012).
2.2. Fungal enumeration
After vortexing for two minutes, tenfold dilution series were
carried out and aliquots of 0.5 ml of each dilution was spread
immediately on Sabouraud chloramphenicol medium (BioRad)
followed by incubation at 25  C for 6e8 days. Fungal colonies were
then counted and levels are expressed as logarithmic scale of col-
ony format units (log10 CFU/cm2) (Hamada and Fujita, 2000). All
experiments were performed in duplicate.
2.3. Fungal isolation and identification
Fungal flora was isolated by direct inoculation on three different
media, Sabouraud agar, Sabouraud agar supplemented with 0.1%
chloramphenicol and Sabouraud chloramphenicol actidione agar.
The plates were incubated at 25  C for 4 weeks, and were observed
every other day for fungal growth. Positive cultures were
subcultured and stored on Sabouraud agar slopes at 4  C. Selected
molds and dermatophytes isolates were identified basing on
macro/microscopic characteristics using Lactophenol Cotton Bleu
(LPCB) slide mount and urease test when necessary. While, yeasts
were identifying using several conventional approaches including
macromorphology, germ tube test, Dalmau plate culture, urease
test, sensitivity to actidione and India ink staining. Further identi-
fication of yeasts was achieved using API 20C AUX system
(Biome rieux).
2.4. Survey and questionnaire
After obtaining the agreement, direct interviews were con-
ducted with the owners of Hammams from which sampling was
performed, in order to seek information about their knowledge of
fungi, their propagation, possibility of fungal survival in Hammams
and the way in which they cause infection. A preliminary investi-
gation was also carried aiming to correlate the data with possible
fungal contamination.
The questionnaire targeted different factors: location of Ham-
mams, opening year, possible renovations, average daily users,
cleaning frequency, cleaned places, and products used for cleaning
and disinfection.
2.5. Data analysis
Fungal a-diversity was calculated in terms of Shannon's di-
P
versity index H 0 ¼  (pi  ln pi) and evenness (E ¼ H'/log S) ac-
cording to sampling position, nature and localization of Hammams
where species richness (S) is the total number of identified fungal
species and pi correspond to the proportion of individuals of total
species represented by ith species (Magurran, 2004). Similarities
between molds and yeasts assemblages were compared using a
Student's t-test with a ¼ 0.05.
Response frequencies were calculated as the percentage of re-
spondents to the total of surveyed Hammam's owners. Pearson's
Chi-squared test (c2) was performed to test the statistical signifi-
cance of differences between groups of each surveyed question.
Analysis of covariance (ANCOVA) was computed to test the ef-
fect of factors mean opening year, average customer, cleaning fre-
quency, locality, nature, sampling position and their interactions on
fungi load recovered from Hammams. In addition, analysis of
variance (ANOVA) was performed testing the effect of sampling
position and surveyed Hammams on fungal load. Multiple com-
parisons of means with Tukey's post hoc analysis were conducted
to distinguish homogeneous and heterogeneous groups among
different variables. Descriptive statistics (mean, standard deviation
and quartiles) were given as box plots to describe the effect of
sampling position, nature and localization of Hammams on total
and specified fungal loads. All computations were performed using
XLSTAT (Addinsoft's, 2014).
3. Results and discussion
3.1. Fungal load and factor affecting their interaction
All studied Hammams yielded positive results for fungal
contamination, which is consistent with conditions of fungus
growth in particular physical environmental factors including hu-
midity and temperature which considering determinants not only
for growth initiation, but to stimulate fungi colonization (Kavanagh,
2011).
The total fungal load depending on sampling sites indicate
highly significant difference (p ¼ 0.003) (Table 1), showing that the
doors and marble massage platforms had the highest rates of
 L. Benammar et al. / International Biodeterioration & Biodegradation 117 (2017) 115e122 117

Table 1 sites (Table 1). Doors, floors and walls exhibited similar mean loads
Outcomes of ANOVA testing the effect of sampling site and surveyed Hammams on varying between 1.76 ± 1.85 and 2.52 ± 1.31 log10 CFU/cm2 (Fig. 1),
fungal load (*: significant, **: highly significant, ***: very highly significant, NS: no
significant).
compared to massage platforms (0.72 ± 1.01 log10 CFU/cm2).
Regarding mold loads, no significant differences (p > 0.05) were
Source Variable Df SS F P Sig reported among sampling sites, which exhibited mean rates of
Yeast load Hammam 9 126.66 16.97 <0.0001 *** 1.56 ± 0.83, 1.58 ± 0.52, 2.01 ± 1.14 and 1.78 ± 0.56 log10 CFU/cm2 for
Sampling site 3 32.67 13.13 <0.0001 *** floors, walls, massage platforms and doors respectively (Fig. 1). Poor
Interaction 27 140.07 6.26 <0.0001 ***
hygiene may increase the fungal load on doors and walls. Based on
Mold load Hammam 9 29.99 7.55 <0.0001 ***
Sampling site 3 2.65 2.00 0.129 NS our survey, all caldarium are provided with a single opening door
Interaction 27 35.80 3.00 0.001 ** with hand pushing which may lead to a large contamination.
Total fungi load Hammam 9 24.53 4.38 0.001 ** Indeed, it was reported that in 40% of surveyed Hammams, doors
Sampling site 3 9.94 5.33 0.003 **
are cleaned once a week and once per month for other 20%, while
Interaction 27 38.58 2.30 0.008 **
40% remaining baths performed daily cleaning using only water.
The high humidity could cause fungal development thus; a high
load of hygrophilous fungus was detected in lower zones of walls
contamination 2.78 ± 0.73 and 2.53 ± 1.13 log10 CFU/cm2 respec- which suggest the vertical distribution of the fungal flora according
tively (Fig. 1). Besides, floors and walls showed fungal loads of to moisture difference (Hamada and Abe, 2009). Marble massage
1.48 ± 0.92 and 2.21 ± 0.70 log10 CFU/cm2 respectively, which share platforms are places where people perform massage and scrub
common characteristics with all the rest sites. their skin, therefore an area rich in keratin and human filth used as
Analyzing the yeast load revealed highly significant differences nutrient source for fungi (Hamada and Abe, 2009; Lee et al., 2007).
(p < 0.0001) within Hammams even between selected sampling Besides in studied Hammams, cleaning of this zone is often made

Fig. 1. Box plots displaying variation of fungal load (Log10 CFU/cm2) among sampling sites, nature and localization of different Hammams. The average values (C), the horizontal
line within the box indicates the median value, the ends of the box show the 25 and 75% quartiles, and the whiskers show the minimum and maximum values.
118 L. Benammar et al. / International Biodeterioration & Biodegradation 117 (2017) 115e122

with water, which is insufficient to reduce fungal flora. However,
Anaissie et al. (2002) reported that floor cleaning with water reduce
the airborne fungi.
Modeling the factors affecting fungal loads revealed that Ham-
mam's locality was significantly associated with yeasts, molds and
total fungi loads (p ¼ 0.0003, p ¼ 0.003, and p ¼ 0.002) respectively.
The ANCOVA analysis indicated a significant effect (p ¼ 0.002,
p ¼ 0.01, p < 0.0001) respectively, for yeasts, molds and total fungi
according to the variation of seniority of Hammams (Table 1).
Furthermore, the mean age, renovation and restoration can influ-
ence considerably the fungal flora in indoor environments (Fairs
et al., 2010). Similar studies showed that the oldest buildings are
most exposed to fungi contaminants (Bartlett et al., 2004; Sabah,
2011). According to ours findings, Hammams that undergone
renovation were least contaminated by molds. Several studies re-
ported a decrease in fungal indoor concentration after renovation
of mold damages (Lignell et al., 2007; Huttunen et al., 2008).
The statistical analysis showed that the fungi flora are not
significantly related to Hammam's nature and cleaning frequency.
Besides, mold and total fungal rates were more significantly asso-
ciated with locality and number of customer (p ¼ 0.011, p ¼ 0.028)
respectively, whereas the others terms of the model showed no
differences (Table 2).
The load of fungal flora according to the gender users (nature of
Hammams) revealed fluctuating values expressed in log10 CFU/cm2
of 2.42 ± 0.66 for women baths and 2.55 ± 0.24 for baths used
alternately by men and women. Whereas no significant variations
were recorded for the yeasts (1.10 ± 0.64 and 01.48 ± 0.61 log10 CFU/
cm2) or molds (2.09 ± 0.30, and 1.97 ± 0.21 log10 CFU/cm2)
respectively (Fig. 1, Table 2) in both Hammams. The result is
probably due to the limited number of surveyed baths used only by
women (two baths), compared to the eight remaining Hammams
alternately used by women and men.
In the present investigation, popular Hammams presents the
mean reservoirs of yeasts, molds and fungal population with
2.01 ± 0.29, 1.85 ± 0.20 and 2.44 ± 0.45 log10 CFU/cm2 respectively,
compared to lower loads detected in favored district (Fig. 1). The
significant variation among fungal load is assigned to socioeco-
nomic level, reduced standard of living and high level of ignorance
about general hygiene practices, which may indirectly influence
dissemination of fungal infections (Ouaffak et al., 2003; Colbeck
and Lazaridis, 2010). Additionally, Hammams in popular district
are among the oldest, less maintained traditional baths and most of
which have not been restored since their inauguration. They char-
acterized by the presence of cracks inducing fungal contamination,
even if baths are maintained relatively dry (Hamada and Fujita,
1999). Further, the hight number of occupants in restricted and
enclosed place like the caldarium without an efficient ventilation
for all investigated Hamamas was found to be more perseverant in
increasing fungal contamination. Same results were reported for
other public buildings such as schools and libraries (Hayleeyesus
and Manaye, 2014; Hospodsky et al., 2015). In addition to the
availability of a suitable nutrient source, other factors that influence
fungal spore growth, distribution and deposition are moisture,
temperature, pH, humidity and oxygen availability (Elena Aringoli
et al., 2008).
In the present survey, it has been reported that the cleaning of
Hammams is based primarily on sodium hypochlorite which is
mainly used without allowing a sufficient contact time to inhibit
the present microbial flora. In fact, no significance correlation was
noted between cleaning frequency and fungal loads probably to the
counterfeit information declared by owners as a precaution of
municipal hygiene inspection services. Although, sodium hypo-
chlorite is effective against fungal growth even at a low concen-
tration but with sufficient duration exposure (Martyny et al., 2005;
Reynolds et al., 2012). It should be noted that sodium hypochlorite
is easily decomposed by hot water and reacts with organic sub-
strates (Ranco and Mishkin, 2007), which reduces its efficiency and
germicidal effect.
3.2. Fungal isolation and abundance
Nineteen different fungi representing four groups were identi-
fied from surveyed Hammams, including 11 genera of molds, two
dermatophytes, one genus of pseudodermatophytes and five yeast
genera (Table 3).
3.2.1. Yeast group
Eight yeast species appeared frequently, C. albicans, C. glabrata,
C. tropicalis, C. lipolytica, Geotrichum sp., Trichosporon sp., Rhodo-
torula sp. and Cryptococcus sp.
The present study showed that Candida was the most recovered
yeast (62.16%) compared to other genera and C. albicans (35.14%)
was the most dominant species in all studied Hammams (Table 3).
Noting that eight of the surveyed Hammams harboring this species.
Same results have been reported by several authors (Ouaffak et al.,
2003; Goksugur et al., 2006).
The prevalence of C. albicans can be explained by the fact that
traditional baths were occupied by women for a period of about
10 h per day, compared to men (6 h). Indeed about 20% of healthy

Table 2
ANCOVA testing the effect of factors “Mean Opening Year, OY”, “Average Number of Customer, ANC”, “Cleaning Frequency, CF”, “Locality, L”, “Nature, N” and their interactions
on fungal load recovered from Hammams (*: significant, **: highly significant, ***: very highly significant, NS: no significant).

Source Yeast load Mold load Total fungi load

Df SS F P Sig Df SS F P Sig Df SS F P Sig

ANC 1 8.003 4.722 0.035 * 1 5.341 9.110 0.004 ** 1 0.358 0.591 0.446 NS
CF 1 0.624 0.368 0.547 NS 1 0.249 0.425 0.518 NS 1 0.785 1.297 0.261 NS
OY 1 17.414 10.275 0.002 ** 1 4.284 7.307 0.010 * 1 13.297 21.960 <0.0001 ***
L 1 25.456 15.021 0.0003 *** 1 5.810 9.911 0.003 ** 1 6.648 10.979 0.002 **
N 1 0.084 0.050 0.824 NS 1 0.126 0.215 0.645 NS 1 0.001 0.002 0.963 NS
ANC  CF 1 3.924 2.316 0.135 NS 1 0.865 1.476 0.231 NS 1 0.224 0.371 0.546 NS
ANC  OY 1 0.055 0.032 0.858 NS 1 0.428 0.730 0.398 NS 1 2.413 3.985 0.052 NS
ANC  L 1 0.239 0.141 0.709 NS 1 4.099 6.992 0.011 * 1 3.133 5.174 0.028 *
ANC  N 1 38.553 22.749 <0.0001 *** 1 0.539 0.919 0.343 NS 1 0.743 1.227 0.274 NS
CF  OY 1 0.766 0.452 0.505 NS 1 0.490 0.836 0.366 NS 1 0.104 0.173 0.680 NS
CF  L 1 0.258 0.152 0.699 NS 1 0.089 0.152 0.699 NS 1 0.974 1.609 0.211 NS
CF  N 1 3.872 2.285 0.138 NS 1 0.005 0.009 0.926 NS 1 0.097 0.160 0.691 NS
OY  L 1 3.314 1.955 0.169 NS 1 1.084 1.849 0.181 NS 1 0.728 1.202 0.279 NS
ANC  CF  OY 1 4.578 2.701 0.107 NS 1 0.649 1.107 0.298 NS 1 0.316 0.522 0.474 NS
ANC  CF  L 1 1.613 0.952 0.334 NS 1 0.856 1.461 0.233 NS 1 0.003 0.005 0.942 NS
 L. Benammar et al. / International Biodeterioration & Biodegradation 117 (2017) 115e122 119

Table 3
Relative abundances of fungal isolates collected in surveyed Hammams (W: Women, M/W: Men and Women, PoD: Popular district, FaD: Favored district).

Isolated species Nature Localization Overall (%)

W (%) M/W(%) PoD (%) FaD (%)

Molds
Penicillium sp. 32 (23.02) 2301 (45.73) 1525 (62.58) 808 (29.55) 2333 (45.12)
Aspergillus sp. 8 (5.76) 1481 (29.43) 193 (7.92) 1296 (47.40) 1489 (28.80)
Fusarium sp. e 406 (8.07) 405 (16.62) 1 (0.04) 406 (7.85)
Sytalidium dimidiatum 53 (38.13) 299 (5.94) 168 (6.89) 184 (6.73) 352 (6.81)
Scedosporium apiospermum 1 (0.72) 314 (6.24) 13 (0.53) 302 (11.05) 315 (6.09)
Cladosporium sp. 22 (15.83) 140 (2.78) 63 (2.59) 99 (3.62) 162 (3.13)
Microsporum audounii e 37 (0.74) 36 (1.48) 1 (0.04) 37 (0.72)
Mucor sp. 1 (0.72) 9 (0.18) 1 (0.04) 9 (0.33) 10 (0.19)
Alternaria sp. 5 (3.60) 3 (0.06) 4 (0.16) 4 (0.15) 8 (0.15)
Trichophyton mentagrophytes 2 (1.44) 3 (0.06) 2 (0.08) 3 (0.11) 5 (0.10)
Rhizopus sp. e 3 (0.06) 1 (0.04) 2 (0.07) 3 (0.06)
Acremonium sp. e 2 (0.04) e 2 (0.07) 2 (0.04)
Rhizomucor sp. 1 (0.72) 1 (0.02) 1 (0.04) 1 (0.04) 2 (0.04)
Microsporum gypsum 1 (0.72) e 1 (0.04) e 1 (0.02)
Phialophora sp. 1 (0.72) e 1 (0.04) e 1 (0.02)
Trichophytonrubrum 1 (0.72) e 1 (0.04) e 1 (0.02)
Not identified 11 (17.91) 33 (0.66) 22 (0.90) 22 (0.80) 44 (0.85)
Yeasts
Candida albicans 216 (56.99) 484 (33.47) 376 (43.82) 324 (28.72) 700 (35.15)
Candida glabrata 54 (14.24) 215 (14.86) 54 (6.29) 215 (19.06) 269 (13.51)
Candida krusei e 53 (3.66) e 53 (4.70) 53 (2.66)
Candida lipolytica e 53 (3.66) e 53 (4.70) 53 (2.66)
Candida tropicalis 53 (13.98) e 53 (6.17) e 53 (2.66)
Candida sp. e 107 (7.40) 53 (6.17) 54 (4.79) 107 (5.37)
Cryptococcus sp. e 53 (3.66) e 53 (4.70) 53 (2.66)
Geotrichum sp. e 215 (14.86) 161 (18.75) 54 (4.76) 215 (10.79)
Rhodotorula sp. 56 (14.77) 51(3.53) e 107 (9.48) 107 (5.37)
Trichosporon sp. e e 107 (12.47) 54 (4.79) 161 (8.08)
Not identified e 215 (14.86) 54 (6.29) 161 (14.27) 215 (10.79)

asymptomatic women hosted Candida as an indigenous vaginal
microbiota (Kurjak and Chervenak, 2015) and the major factor of
Candida infections compared to men, as for vulvovaginal candidi-
asis (VVC) which affects 75% of women at least one episode in their
lifetime (Schlossberg, 2015). In addition, Candida is a part of the oral
and gastrointestinal human microbiota, counting approximately
40%e71.4% of healthy carriers (Kumamoto, 2011; Ribeiro and
Edvaldo, 2015).
Candida glabrata ranked in second position with (13.15%) fol-
lowed by C. tropicalis (5.26%), C. krusei and C. lipolytica with a
common rate of 2.63% of the total yeast isolates. Recently,
C. glabrata has become a major opportunistic pathogen and etio-
logical agent of superficial or systemic infections after Candida
albicans, especially in immunocompromised patients (Silva et al.,
2012). Current epidemiological data have highlighted increasing
incidence of the non-albicans Candida species such as C. krusei, C.
tropicalis, and C. lipolytica in dissemination of candidiasis (Bolotin-
Fukuhara and Fairhead, 2014).
Other species were isolated, Geotrichum sp. and Trichosporon sp.
with frequency of 10.81% and 8.11%, respectively. The two species
induce uncommon but serious invasive infections exclusively in
immunocompromised patients with hematological malignancies
and severe neutropenia (Girmenia et al., 2005). Similarly, other data
also showed isolation of Rhodotorula sp. (Ara et al., 2004; Hamada
and Fujita, 2000). Formerly regarded non-pathogenic, some species
of Rhodotorula have emerged as opportunistic pathogens having
the capacity to infect immunocompromised patients and even
animals, or to cause fungemia (Wirth and Goldani, 2012). The
presence of this yeast was probably due to her ability to grown on
the soap and human dirt in baths (Hamada and Fujita, 1999).
3.2.2. Mold group
The occurrence of molds and the total frequency of each fungus
in different Hammams are summarized in Table 3. Six fungi groups
appeared frequently and the highest rate was recorded for Peni-
cillium sp. (45.12%), followed by Aspergillus sp. (28.80%), Fusarium
sp. (7.85%), Sytalidium dimidiatum (6.81%), Pseudallescheria boydii
(6.09%) and Cladosporium sp. (3.13%).
According to localization, except Microsporum gypsum, Phialo-
phora sp., Trichophyton rubrum, and Acremonium sp. all mold spe-
cies were common with fluctuating frequencies between favored
and popular districts with a predominance of Aspergillus sp.
(47.40%) and Penicillium sp. (62.58%), respectively. Besides, Ham-
mams used alternately by men and women exhibited the important
rate of Penicillium sp. (45.37%) and Aspergillus sp. (29.43%), while
those used only by women where dominated by Sytalidium dimi-
diatum (38.13%) followed by Penicillium sp. (23.02%).
Molds flora in traditional baths was characterized by a large
number of hydrophilic species like Cladosporium, Fusarium,
Rhizopus, Mucor, Alternaria, probably due to the high humidity in
Hammams. Our findings were in concordance with those reported
by Hamada and Fujita (1999) and Ara et al. (2004). Some molds
isolates were either thermotolerant which grow well up to 50  C
such as some Aspergillus (A. candidus A. niger, A. falvus, A. terreus),
Penicillium and Rhizopus species or thermophilic like Aspergillus
fumugatus, some Rhizomucor and Mucor species. Additionally, it has
been shown that most of the thermophilic and thermotolerant
fungi are hydrophilic (Johri et al., 1999).
A low frequency (0.98%) of dermatophytes was reported in our
study compared to all isolates. This could be due to the presence of
chlorine residues which inhibits their growth (Moriello et al., 2013),
to the seasonal variation of which the number of dermatophytes
decrease during winter and spring (Lee et al., 2014; Gniadek et al.,
2005), coinciding with the period of our study (February and May),
or to the use by Algerian women of some products having anti-
fungal properties against dermatophytes or pathogenic fungi such
120 L. Benammar et al. / International Biodeterioration & Biodegradation 117 (2017) 115e122

as henna and cosmetics (Gozubuyuk et al., 2014; Suleiman and
Mohamed, 2014). On the other hand, the viability of the low con-
tent of dermatophytes in Hammams is possibly related to dead cells
from exfoliation or hair debris of the users that constitutes a source
of keratin to these keratinolytic fungi (Ouaffak et al., 2003;
Yenisehirli et al., 2012).
The difference in total frequency of molds (72.25%) compared to
yeasts (27.75%) is may due to the fact that two of total surveyed
Hammams appeared with no yeasts growth. The lack of yeasts
could probably due to the sampling method or as a result of anti-
fungal activity exercised by some fungi (ex. Aspergillus flavus) pre-
senting a significant activity against Candida albicans and Candida
tropicalis (Bhattacharyya et al., 2013). On the other hand, isolation
of yeasts was more difficult than for filamentous fungi which
growth consists of hyphae causing sharp rise limiting development
of yeasts in the medium (Pitt and Hocking, 2009).
3.3. Fungal diversity and evenness
3.4. Level of knowledge about fungi in Hammams
Information's relating to the level of knowledge about fungi in
the studied populations (Hammam's owners) are summarized in
Table 5. The analysis of questionnaire data by the Pearson's c2 test
showed that the respondent groups were significantly differed
between responses level of information about fungi (c2 ¼ 10.41,
p ¼ 0.005), with 80% of respondents have limited information, 20%
have no idea and none owners showed extensive or detailed
knowledge about fungi.
As for fungal propagation methods, the Chi-square test revealed
a significant difference (c2 ¼ 3.60, p ¼ 0.011), where only 10% of
surveyed subjects present some information (Table 4). No differ-
ences were noted for education level (c2 ¼ 0.80, p ¼ 0.670),
knowledge about possible fungal growth in Hammams (c2 ¼ 0.40,
p ¼ 0.527) and the possibility of contracting diseases via fungi
(c2 ¼ 1.60, p ¼ 0.206). Hammam's owner were mainly aged be-
tween 20 and 40 years (50%), with 40% belonging to 40e60 age

Yeast population isolated from all Hammams had the greater

Table 5
and significant (p ¼ 0.007) diversity values (H' ¼ 2.03 bits, S ¼ 25) Main investigation parameters of Hammam's owners about their knowledge on
followed by molds with (H' ¼ 1.75 bits, S ¼ 11) (Table 4). fungi.
Generally, The mean number of isolated species, Shannon
Variable % c2 p-value
indices and evenness depending on sampling sites, nature and
A. Age of the owners (years) 2.60 0.272
localization of Hammams were almost similar for yeast and mold
20-40 50
mycobiotas (Table 4), except for the air (H’ ¼ 0, S ¼ 1). These results 40-60 40
indicate similarity with respect to the number of isolates found in > 60 10
them. B. Education level 0.80 0.670
The present results showed that molds community was un- Illiterate 40
High school 40
even due to the consistent high predominance of Penicillium sp. University graduate 20
(45.12%) and Aspergillus sp. (28.80%) respectively. In fact, the two C. Information about fungi 10.41 0.005
species were noticed to be prevalent not only in bathrooms and None 20
Hammams, but also in most indoor environments sharing favor- Limited information 80
Much information 00
able factors for their development (Borrego et al., 2010;
D. Fungal dissemination ways 6.40 0.011
Ghahfarokhi et al., 2014). These funguses are known as patho- Yes 10
genic responsible for severe respiratory disorders (Allergic rhinitis No 90
to asthma) and mycotoxins producers affecting drastically human E. Possible growth of fungi in Hammams 0.4 0.527
health (Nielsen, 2003; Bundy et al., 2009; Ruga et al., 2015). Yes 40
No 60
Furthermore, moderate diversity for molds was noted in the sur-
F. Might fungi cause diseases? 1.6 0.206
veyed Hammams, probably due to the lower number of isolates Yes 70
and to the greater number of rare species presents only in limited No 30
number of studies Hammams.

Table 4
Species richness and diversity of identified fungi in surveyed Hammams (N: number of fungal isolates, S: species richness, H: Shannon's index, E: evenness).

Source Molds Yeasts

N S N/S H0 E N S N/S H0 E

Hammams H1 71 10 7.10 1.05 0.46 54 1 54 0 e

H2 64 7 9.14 1.08 0.55 376 4 94 1.55 1.12
H3 628 8 78.50 0.88 0.42 108 1 108 0.69 e
H4 1108 10 110.80 1.33 0.58 428 7 61.14 1.91 0.98
H5 206 11 18.73 1.49 0.62 0 0 0 e e
H6 1063 7 151.86 1.26 0.65 215 2 107.5 0.69 1.00
H7 68 10 6.80 1.53 0.66 268 2 134 0.50 0.72
H8 1286 5 257.20 0.65 0.40 0 0 0 e e
H9 388 8 48.50 1.17 0.56 323 2 161.5 0.45 0.65
H10 289 9 32.11 1.37 0.62 214 3 71.33 1.04 0.95
Sampling position Floor 1270 17 74.70 0.92 0.32 216 3 72.00 1.04 0.95
Wall 1040 16 65.00 1.71 0.62 107 2 53.50 0.69 1.00
Door 1007 14 71.93 1.54 0.58 1286 8 160.75 1.75 0.84
Platform 1583 13 121.77 1.23 0.48 324 3 108.00 0.87 0.79
Air 271 11 24.64 1.11 0.46 53 1 53.00 0.00 e
Nature Women 139 17 8.18 1.85 0.65 323 3 107.67 0.87 0.79
Men/women 5032 22 228.73 1.72 0.56 1663 10 166.30 1.85 0.80
Localization Favored district 2734 22 124.27 1.79 0.58 1127 10 112.7 0.89 2.05
Popular district 2437 21 116.04 1.26 0.41 856 7 122.28 0.84 1.63
Over all 5171 25 206.84 1.75 0.54 1986 11 180.54 2.03 0.85
 L. Benammar et al. / International Biodeterioration & Biodegradation 117 (2017) 115e122
class and only 10% were above 60 years. For the respondent's ed-
ucation level, 40% are either high school graduates or illiterate and
20% were university graduates.
Regarding the results of interviewed subjects, it was found that
the owners have presented limited and insufficient information
about fungi and it was obvious since most of the questioned sub-
jects (80%) had restricted level of education, as well as lack of
commitment and indifference toward hygiene rules applied in
Hammams, which prevented having cognizance about the subject.
4. Conclusions
Algerian traditional baths are a highly frequented public places,
considering for a long time as a cultural and historical heritage.
However, such closed, hot and wet places with poor hygienic rules
may pose a serious risk factor by spreading fungal pathogens. Re-
sults of this study showed high fungal load in the surveyed Ham-
mams with varying differences among sampling parts, and even a
large diversity of fungal species, including human pathogens.
Moreover, the findings revealed that average number of customers,
mean opening year, and locality are the most involved parameters
that significantly influence the fungal loads in traditional public
baths. Further studies are clearly needed to better understand and
investigate other relevant parameters that influence fungal di-
versity in Hammams and to suggest control methods against fungal
dissemination in such indoor environment which imposing a real
threat on public health.
Conflict of interest
The authors declare that they have no conflicts of interest.
Funding sources
This research did not receive any specific grant from funding
agencies in the public, commercial, or not-for-profit sectors.
Acknowledgements
We gratefully acknowledge laboratory staff of microbiology at
the Department of Veterinary Sciences (University of Batna 1) for all
facilities provided during carrying out this study.
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