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International Biodeterioration & Biodegradation 60 (2007) 231–237

Aeromycological study in the Cathedral of

Santiago de Compostela (Spain)
M.J. Airaa, V. Jatob, A.M. Stchigelc, F.J. Rodrı́guez-Rajob,, E. Piontellid
Department of Botany, Faculty of Pharmacy, University of Santiago de Compostela, Campus South, Santiago de Compostela, Spain
Department of Plant Biology and Soil Sciences, Sciences Faculty Ourense, University of Vigo, Campus As Lagoas, Spain
Microbiology Unit, Medicine and Health Sciences Faculty, Rovira y Virgili University, Tarragona, Spain
Mycology Chair, Medicine School, Valparaı´so University, Valparaı´so, Chile

Received 26 June 2006; received in revised form 27 January 2007; accepted 28 February 2007
Available online 23 May 2007


A study of airborne fungi was carried out in the architectural complex of the Cathedral of Santiago de Compostela (Spain) during
2002, by using viable volumetric sampling methods. This resulted in a total of 35 identified taxa, of which the most abundant were:
Alternaria, Aspergillus, Cladosporium and Penicillium. Sampling was completed with data from the outdoor atmosphere and swab
samples in specific places.
In general there were no statistically significant indoor/outdoor differences and in both cases the highest CFU m-3 were obtained
during the spring-summer. Similar relatively low numbers of the same fungi were likewise detected at different points in the Cathedral
nave, while up to nearly 6500 CFU m-3 were recorded in the Corticela Chapel. The study of intradiurnal levels carried out in the
Cathedral nave reveals greater abundance of fungal concentrations at 13:00 h, the moment of massive influx of visitors in the Cathedral,
with 406 CFU m-3 compared to the 380 CFU m-3 sampled at 9:00 h and the 350 CFU m-3 at 21:00 h. The whole investigation is the first
study of the atmospheric fungal content of the Cathedral of Santiago de Compostela.
r 2007 Elsevier Ltd. All rights reserved.

Keywords: Aerobiology; Biodeterioration; Fungal spores; Santiago of Compostela Cathedral; Volumetric sampling

1. Introduction goal of studies of the mycobiota carried out in heritage

Different microorganisms such as fungi, algae and Studies on biodeterioration deal with different topics
bacteria may have a negative effect on the preservation and substrates. There is great diversity in relation to the
of artistic-historical heritage, especially when microclimatic subjects of such studies since they encompass any type of
conditions favour their development. Numerous museum work of art (Guglielminetti et al., 1994; Gorbushina and
complexes therefore employ systematic temperature and Palinska, 1999; Pitzurra et al., 1999; Maggi et al., 2000), as
humidity monitoring, with the aim to prevent or slow down well as the materials of the buildings (Caretta and Piontelli,
their growth (De Nuntiis et al., 2004). 1998; Petushkova and Kandyba, 1999). However, despite
Some authors also report interaction between micro- the interest of this type of study, those related to fungal
scopic fungi and arthropods on the surface of wall biodeterioration are not very common; in the case of the
paintings, which increases their alteration (Hoffland Cathedral of Santiago de Compostela, no previous studies
et al., 2004). These biological agents affect not only the have been conducted.
aesthetical appearance but also the structure of materials. The main objective of this study was therefore to
Contributing to their preservation therefore is the main ascertain the mycobiota atmospheric content of the city’s
most important architectural complex, which forms part
Corresponding author. Tel.: +34 988387054; fax: +34 988387001. of its World Heritage as pointed out by UNESCO, to
E-mail address: (F.J. Rodrı́guez-Rajo). identify the agents causing the proliferation of these

0964-8305/$ - see front matter r 2007 Elsevier Ltd. All rights reserved.
232 M.J. Aira et al. / International Biodeterioration & Biodegradation 60 (2007) 231–237

microorganisms and its favouring factors. This will pave same conditions as the previous ones, with the exception that the
Sabouraud was replaced by MEA and oat agar (OA).
the way for a proposal regarding any necessary conserva-
The influence of the high quantity of people visiting the Cathedral on
tion work to the building, which is showing some evidence Sundays was studied, since religious services may be attended by more
of alteration caused by fungi in certain areas. than 5000 people. To evaluate whether this variable influences fungal
Indirectly, this type of study may have public health content, every fourth Sunday, air in the centre of the Nave (Point 5) and
applications, since the calculation of atmospheric fungal outdoors was sampled onto Sabouraud agar using the BPC. The samples
content and the identification of certain human pathogens were conducted when low numbers of visitors are in the Cathedral
(09.00 h), in the moment of the high influx of visitors (13.00 h) and in an
can show whether an atmosphere is healthy or not and hour of the day with medium people presence (21.00 h), so as to assess its
point to potential allergy risks (Terr, 2004). In this regard, impact on the data obtained.
fungi such as Stachybotrys are a usual component of the Other seasonal samplings (Spring: April 21, Summer: August 11,
mycobiota inside buildings with water-damaged walls Autumn: November 3 and Winter: December 29, January 27, February
1st) was carried out (Fig. 1) inside the Cathedral chapels (Point a:
(Terr, 2001).
Corticela and Point b: Santı́simo) and inside the Museum’s exhibition
rooms (Point c: Library and Point d: Goya), where possible alterations
2. Material and methods produced by fungi were detected, to determine the particular atmospheric
fungal content in each area. In this case both volumetric sampling systems
This study was carried out during the year 2002 in the main were used, with Sabouraud agar as before as culture medium. Finally,
monumental complex of Santiago de Compostela (Spain), made up of swabbing was used to obtain samples directly from the walls inside of the
the Cathedral and Museum. Chapel of Corticela and the Chapel of Santı́simo, the sides of the ‘‘Pórtico
Atmospheric spore samples were collected by using two viable de la Gloria’’ (which is located in the Cathedral nave) and from different
volumetric sampling systems. These are a six-stage Andersen sampler artistic pieces on display inside the Museum’s exhibition rooms Library
(A) and a single-stage Burkard portable air sampler for agar plates (BPC) and Goya.
including 1 sampling plate. The sampling lasted 10 min been the collectors The culture plates obtained were incubated at 25 1C during 7 d, after
placed 1 m above ground level. The results were expressed in CFU m-3 which the colonies were counted and isolated. Simultaneously, in all of the
and the positive-hole correction in both samplers was applied (Mehta et samples, temperature and humidity was measured using a weatherlink
al., 1996). Random swab samples were also collected, to identify the meteorological station; days with rainfall were also recorded due to the
fungal types present on certain works of artistic value or ‘‘areas’’ showing possible influence on the outdoor atmosphere’s fungal content.
fungal growth. The entire methodology used in the study was explained in Finally, we applied Scheffé’s test to study the homogeneity of the
detail by Aira et al. (2005). The preparation of culture media was populations under study (Wassertheil-Smoller, 2004), which would
conducted following the instructions of Hoog et al. (2000). enabled us to identify the presence of differences in the quantity of
With the aim to ascertain the fungal quantity and diversity throughout mycobiota content between the sampling points. Spearman’s correlation
the year, sampling was carried out every four weeks at five points in the test was used to find a possible relationship between the fungal levels and
Cathedral nave, four at the extremes (identified as Point 1: Quintana, meteorological factors (temperature and humidity) or visitors affluence.
Point 2: Azabacheria, Point 3: Portico and Point 4: Platerias) and one at
the centre (Point 5: Nave centre) of the Latin-cross ground plan. Sampling
3. Results
was also carried out outside the Cathedral (Point 6: Outdoors) as a
reference (Fig. 1). Friday was chosen as a representative day of the
Cathedral’s normal activity, i.e. without a massive influx of visitors that 3.1. Quantitative data
could affect atmospheric fungal content. At each sampling time and
location, single samples were taken using the BPC and commercial The total number of colony forming units in all of the
Sabouraud agar amended with chloramphenicol to control bacteria.
samples taken in the Cathedral nave (38,137 CFU) was
To evaluate efficiency of Sabouraud agar culture media in comparison
with malt extract agar (MEA) and oatmeal agar (OA), sampling was higher than that of outdoor air (20,509 CFU). However,
carried out in the central area of the nave (Point 5: Nave centre) under the despite this apparent predominance of fungi inside the
Cathedral (Table 1), when applying Scheffé’s test no general
significant differences (po0.05) were revealed when com-
paring each sampling point inside the nave with the outside.
In general, the results obtained throughout the year
inside the nave of the Cathedral show that the maximum
concentration is recorded during spring and summer
months (with the exception of sampling in April), decreas-
ing considerably in the first 3 months of the year. The

Table 1
Mean of CFU and range of counts for each sampling point in the nave of
the Cathedral

Point 1 Point 2 Point 3 Point 4 Point 5 Outdoors

Quintana Azabacherı́a Pórtico Platerı́as Nave
Fig. 1. Sampling site points in the nave Cathedral (1—Quintana, 2—
Azabacherı́a, 3—Pórtico, 4—Platerias, 5—Nave Centre, 6—Outdoors) Range 135–560 145–615 70–610 85–570 105–560 55–540
and seasonal sampling site points (a—Corticela Chapel, b—Santı́simo Mean 327 350 388 339 357 309
Chapel, c—Library Museum room, d—Goya Museum room).
M.J. Aira et al. / International Biodeterioration & Biodegradation 60 (2007) 231–237 233

700 1 2 3 4 5 Out







Jan 25 Feb 22 Mar 22 Apr 10 May 17 Jun 14 Jul 12 Aug 9 Sep 6 Oct 4 Nov 1 Nov 29 Dec 27

Fig. 2. Representation of number of CFU m-3 throughout the year in the Cathedral nave sampling points 1—Quintana, 2—Azabacherı́a, 3—Pórtico, 4—
Platerias, 5—Nave Centre and 6—Outdoors.

minimum values were obtained in April at all sampling Table 2

points, with the exception of point 2, which was recorded in Number of CFU m-3 sampled at different hours of the day (09.00, 13.00
and 21.00 h) indoor (in the sampling point 5: Nave Centre) and outdoor
The most marked differences among the five sampling Indoor Outdoor
points in the nave were recorded in September (points 1
and 2), January (points 3 and 4) and November (points 3 09.00 13.00 21.00 09.00 13.00 21.00
and 5); comparing the indoor and outdoor atmospheres, 27 Jan 385 510 550 255 255 465
the greatest values occurred in January and August with 1 Feb 530 575 540 550 85 115
amounts greater than 300 CFU m-3 (Fig. 2). 24 Mar 505 530 380 390 130 295
In relation to culture media, the most effective medium 21 Apr 195 220 215 105 95 320
19 May 245 455 260 355 315 245
from a quantitative point of view was MEA, in which a
16 Jun 460 495 495 575 505 530
mean of 380 CFU m-3 was recorded, followed by Sabour- 14 Jul 580 630 575 650 645 610
aud agar with 357 CFU m-3 and finally OA with 11 Aug 455 490 410 445 515 480
308 CFU m-3. The greatest differences in relation to culture 8 Sep 470 465 275 530 205 535
media were obtained between OA and MEA, although they 6 Oct 285 395 320 470 505 430
3 Nov 405 95 165 460 185 480
were not statistically significant. Nevertheless, the highest
1 Dec 210 200 145 435 275 260
individual value corresponded to the sampling carried out 29 Dec 210 220 215 335 255 245
on March 24th with Sabouraud (560 CFU m-3).
Mean 380 406 350 427 305 385
The study of intradiurnal levels carried out in the
Cathedral nave reveals greater abundance of fungal
concentrations around midday (at 13:00 h) in the moment
of the high influx of visitors, with 406 CFU m-3, compared Cathedral around mid-day, despite this differences were
to the 380 CFU m-3 at 09:00 h and 350 CFU m-3 at 21:00 h. not significant according to Spearman’s correlation test.
The levels recorded in the early morning are higher In the seasonal sampling (Table 3) carried out inside the
outdoors, with 427 CFU m-3 at 09:00 h compared to the Cathedral chapels and inside the two Museums rooms,
305 CFU m-3 sampled at 13.00 h and 385 CFU m-3 at the number of colony forming units was higher using the
09.00 h (Table 2). In the nave of the Cathedral, the values Andersen sampler, and the indoor levels were higher than
obtained at 13:00 h were higher than those at 09:00 h in 9 the outdoor ones in both cases. This fact should be caused
of the 13 sampling days. The highest differences, greater as a six-stage sampler achieves a better separation of spores
than 100 CFU m-3, were obtained in May, January and than a one-stage sampler, and also because it sampled a
October. The values recorded at 13:00 h were likewise larger volume of air. Some unexpected results such us, the
higher than those at 21:00 h in 10 of the 13 sampling days. winter BPC count inside Library Museum room were lower
The most marked differences (greater than 100 CFU m-3) than outdoors, the summer BPC count in the Goya
occurred in May, September and March. Museum room were higher than the Andersen count and
The comparison of indoor/outdoor results shows that the outdoor level in autumn were higher than inside the
the differences are greatest at 13:00 h, when the indoor Santı́simo Chapel, Library Museum room and Goya
concentrations are very high in relation to the outdoor Museum room values, could be caused by the fact only
ones. Even in the case of those days when the outdoor that a single sample has been taken for each season.
concentrations are highest, the difference decreases be- The highest levels inside the two chapels were obtained
tween 9:00 and 13:00 h. These differences seem to indicate at different times of year, as it was the case of the two
the influence of the massive influx of visitors in the Museum rooms, while outdoors they were recorded during
234 M.J. Aira et al. / International Biodeterioration & Biodegradation 60 (2007) 231–237

Table 3 Cladosporium, which includes species such as Cladosporium

Number of CFU m-3 obtained in the seasonal sampling of different cladosporioides (Fresen.) G.A. de Vries, Cladosporium
Cathedral Chapels and rooms
avellaneum G.A. de Vries, Cladosporium elatum (Harz)
Spring Summer Autumn Winter Annual total Nannf., Cladosporium oxysporum Berk. and M.A. Curtis,
among others, and herbarum-type Cladosporium, which
Corticela Chapel encompasses Cladosporium herbarum (Pers.) Link, Clados-
Andersen 2943 1547 1470 509 6469
porium sphaerospermum Penz. and Cladosporium macro-
BPC 545 505 335 484 1869
Temperature 13 21 21 12 13 carpum Preuss (Mediavilla et al., 1995).
Humidity 65 58 66 76 Within the Aspergillus genus, the most abundant species
was Aspergillus fumigatus Fresen., representing up to 28%
Santı´simo Chapel
Andersen 539 373 360 966 2238 of the outdoor fungi count, while Penicillium purpurogenum
BPC 480 525 300 490 1795 Stoll was dominant within its genus, with a maximum
Temperature 17 21 25 14 representation of 48% (Table 6).
Humidity 57 64 60 80 In relation to the results obtained by swabbing the inside
Library Museum room walls of the Santı́simo Chapel, we identified Arthrobotrys
Andersen 733 266 348 187 1534 oligospora Fresen., Aspergillus nidulans (Eidam) G. Winter,
BPC 460 175 215 270 1120 Aspergillus niger Tiegh., Aspergillus versicolor (Vuill.)
Temperature 15 22 22 16
Tirab., Aureobasidium pullulans (de Bary) G. Arnaud,
Humidity 56 61 54 65
Curvularia lunata (Wakker) Boedijn, Penicillium aspergil-
Goya Museum room loides Rudakov, P. purpurogenum Stoll and Trichoderma
Andersen 255 113 227 304 899
viride Pers. in green and brownish-grey patinas, while
BPC 335 210 185 405 1135
Temperature 15 23 22 17 A. pullulans(de Bary) G. Arnaud and Cunninghamella
Humidity 54 55 50 63 elegans Lendn. formed blackish powdery patinas on the
sides of the ‘‘Pórtico de la Gloria’’ and P. purpurogenum
Andersen 235 180 540 96 1051 Stoll colonised several religious pieces of great artistic value
BPC 5 165 390 360 920 in the inside walls of the Corticela Chapel.
Temperature 21 23 17 14 Different species of Penicillium, A. fumigatus and Absidia
Humidity 45 53 57 66 corymbifera (Cohn) Sacc. and Trotter were identified with
The values of the meteorological data during the samples were show. the same technique in several display cabinets inside the
Cathedral Museum’s library room; A. corymbifera (Cohn)
Sacc. and Trotter was also isolated from a powdery sample
from the Museum’s Choir Room. In the Goya Museum
autumn, probably due to the influence of local mycobiota room, where mainly tapestries are on display, there were
content and meteorological variables. Temperature in the also different species of Penicillium and Chaetomium,
indoor atmospheres sampled oscillated between 12 and A. fumigatus Fresen., A. versicolor (Vuill.) Tirab., C.
15 1C and humidity between 54% and 80%. cladosporioides (Fresen.) G.A. de Vries and Scopulariopsis
Spearman correlation test was applied to ascertain the brumptii Salvanet-Duvalin.
association degree between fungal levels and temperature
and humidity, revealing a positive correlation with 4. Discussion
humidity (R ¼ 0.475) and a negative one with temperature
(R ¼ 0.502), with 99% significance in both cases. The study of the Cathedral nave’s fungal representation
reveals the existence of mean fungal levels greater than
3.2. Qualitative data 325 CFU m-3 at all sampling points. The application of
Scheffé’s statistical test to the results indicates that there
A total of 35 taxa were identified and the greatest fungal are no significant differences, but to confirm this result a
diversity was obtained in the samples taken in the larger study by means a higher number of plates exposed
Cathedral nave, with 31 taxa identified, followed by should be done, as microorganisms in indoor air are not
the outdoor atmosphere with 23 taxa and the Museum uniformly dispersed, and samples taken at the same
samples, in which a total of 14 taxa were identified location within minutes of each other could be markedly
(Table 4). different. At all of the sampling points the mean levels were
The four most abundant genera, regardless of the higher than those recorded outdoors (309 CFU m-3),
sampling point, were: Alternaria, Aspergillus, Cladosporium although such differences were not significant.
and Penicillium, with the first three dominant in the nave On the other hand, inside the Corticela and Santı́simo
while Alternaria stands out slightly outdoors (Table 5). chapels, using the data obtained with the BPC (which is,
The most abundant Alternaria species was Alternaria therefore, comparable to that of the similar points in the
alternata (Fr.) Keissl., while in the case of Cladosporium, nave), the mean values are higher, and surpass those of the
there was a codominance between cladosporioides-type outdoor atmosphere.
M.J. Aira et al. / International Biodeterioration & Biodegradation 60 (2007) 231–237 235

Table 4
Taxa identified by using volumetric sampling methods in the Cathedral nave, Chapels and Museum rooms, and outside

Cathedral Nave Cathedral Chapels Museum rooms Outdoors

Corticela Santı́simo Library Goya

Acremonium sp. + + +
Alternaria sp. + + + + + +
Arthrinium phaeospermum (Corda) M.B. Ellis + + + +
Aspergillus candidus Link + +
Aspergillus flavus Link + + +
Aspergillus fumigatus + + + + +
Aspergillus niger Fresen. + + +
Aspergillus oryzae (Ahlb.) E. Cohn +
Aspergillus terreus Thom + +
Aspergillus versicolor (Vuill.) Tirab. +
Aspergillus sp. + + + + + +
Beauveria bassiana (Bals.-Criv.) Vuill. + +
Botrytis cinerea Pers. + +
Chaetomium globosum Kunze + +
Chaetomium indicum Corda + + +
Cladosporium sp. + + + + + +
Cylindrocarpon sp. +
Microsphaeropsis pseudaspera B. Sutton +
Mucor mucedo L. + + +
Paecilomyces lilacinus (Thom) Samson + + + +
Paecilomyces variotii Bainier + + + + +
Penicillum aurantiogriseum Dierckx + +
Penicillium citrinum Sopp +
Penicillium crustosum Thom + +
Penicillium funiculossum Thom +
Penicillium glabrum (Wehmer) Westling + +
Penicillium pinophillum Thom + +
Penicillium purpurogenum Stoll + + + + + +
Penicillium spinulosum Thom +
Penicillium thomii Maire + +
Penicillium sp. + + + + + +
Rhizopus stolonifer (Ehrenb.) Vuill. + + + + +
Trichoderma sp. +
Stachybotrys atra Corda +
viride Pers. +

Table 5
Number of colonies identified of the four most abundant genera

Cathedral Nave Cathedral Chapels Museum rooms Outdoors

Corticela Santisimo Library Goya

Alternaria 47 15 7 18 12 53
Aspergillus. 185 114 40 33 19 74
Cladosporium 580 12 17 39 25 512
Penicillium 401 81 59 48 28 186

Applying Scheffé’s test to the results obtained with the microclimate that is especially suitable for fungal develop-
Anderson method reveals the existence of significant ment. In fact, there is evident deterioration in some
differences between those inside the Corticela Chapel and wooden beams inside of the Chapel of Corticela, while
those from the other sampled rooms and with those from there is an abundance of patinas on the dampest walls
outdoors, thereby indicating differential behaviour of its indoors of the Santı́simo Chapel. In view of the meteor-
aeromycobiota. The levels attained indoors the Corticela ological data, temperature does not seem to pose a risk to
Chapel are higher than 1400 CFU m-3 in three of the four the artistic material’s preservation, unlike humidity, whose
seasonal samples. These quantitative differences may be values exceed recommended levels, especially indoors the
due to the fact that the physical isolation favours a Cathedral’s chapels.
236 M.J. Aira et al. / International Biodeterioration & Biodegradation 60 (2007) 231–237

Table 6
Representation of the most abundant species (in %)

Cathedral Nave Cathedral Chapel Museum rooms Outdoors

Corticela Santisimo Library Goya

Number of A. fumigatus isolations 29 1 6 6 3 21

Number of Aspergillus sp. isolations 185 114 40 33 19 74
A. fumigatus (%) 16 1 15 18 16 28
Number of P. purpurogenum isolations 47 1 11 23 7 41
Number of Penicillium sp. isolations 401 81 59 48 28 186
P. purpurogenum (%) 12 1 19 48 25 22

Similar values to those of this study were detected in investigation and further research, by means other culture
Rome’s State Archives, where about 300 CFU m-3 were media and a higher number of plates exposed, should be
recorded, the most predominant genera being Cladospor- conducted to complete and verify this results with the aim
ium in winter and Penicillium in summer, along with others to generate more precise data for the preservation of
commonly found throughout the year such as Alternaria, artistic-historical heritage.
Aspergillus and Chaetomium (Maggi et al., 2000). On the
other hand, in India, a study carried out with an Andersen Acknowledgements
trap in several Delhi libraries showed a much higher level,
since the Cladosporium genus alone amounted to This study was subsidised by the Department of Culture,
12,322 CFU m-3. Other important genera were Aspergillus, Social Communication and Tourism of the Xunta de
Penicillium and Alternaria (Singh et al., 1995). Galicia (PGIDT01PAT38301PR).
Qualitatively speaking, the taxa identified in Santiago
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