Rev Iberoam Micol. 2017;34(2):99–105
Revista Iberoamericana
de Micología
www.elsevier.es/reviberoammicol
Original article
Fungal monitoring of the indoor air of the Museo de La Plata
Herbarium, Argentina
Andrea C. Mallo a,e,∗ , Lorena A. Elíades b,f , Daniela S. Nitiu a,f , Mario C.N. Saparrat b,c,d,f
a
Cátedra de Palinología, Facultad de Ciencias Naturales y Museo, Universidad Nacional de La Plata, Argentina
Instituto de Botánica Carlos Spegazzini, Facultad de Ciencias Naturales y Museo, Universidad Nacional de La Plata, Argentina
Instituto de Fisiología Vegetal, Argentina
d
Cátedra de Microbiología Agrícola, Facultad de Ciencias Agrarias y Forestales, Universidad Nacional de La Plata, Argentina
e
Comisión de Investigaciones Científicas de la Provincia de Buenos Aires (CIC-PBA), Argentina
f
Consejo Nacional de Investigaciones Científicas y Tecnológicas (CONICET), Argentina
b
c
a r t i c l e
i n f o
Article history:
Received 7 November 2014
Accepted 11 May 2016
Available online 15 February 2017
Keywords:
Biological collections
Fungal spores
Indoor airborne fungi
La Plata Herbarium
Viable and non-viable methods
a b s t r a c t
Background: Biological agents, such as fungal spores in the air in places where scientific collections are
stored, can attack and deteriorate them.
Aims: The aim of this study was to gather information on the indoor air quality of the Herbarium of
Vascular Plants of the Museo de Ciencias Naturales de La Plata, Argentina, in relation to fungal propagules
and inert particles.
Methods: This study was made using a volumetric system and two complementary sampling methods: (1) a non-viable method for direct evaluation, and (2) a viable method by culture for viable fungal
propagules.
Results: The non-viable method led to ten spore morphotypes being found from related fungal sources.
A total of 4401.88 spores/m3 and 32135.18 inert suspended particles/m3 were recorded. The viable
method led to the finding of nine fungal taxa as viable spores that mostly belonged to anamorphic forms
of Ascomycota, although the pigmented yeast Rhodotorula F.C. Harrison (Basidiomycota) was also found.
A total count of 40,500 fungal CFU/m3 air was estimated for all the sites sampled.
Conclusions: Both the non-viable and viable sampling methods were necessary to monitor the bio-aerosol
load in the La Plata Herbarium. The indoor air of this institution seems to be reasonably adequate for the
conservation of vascular plants due to the low indoor/outdoor index, low concentrations of air spores,
and/or lack of indicators of moisture problems.
© 2016 Asociación Española de Micologı́a. Published by Elsevier España, S.L.U. All rights reserved.
Muestreo de los hongos del aire en el interior del Herbario del Museo
de La Plata, Argentina
r e s u m e n
Palabras clave:
Colecciones biológicas
Esporas fúngicas
Aire interior
Herbario La Plata
Muestreos viable y no viable
Antecedentes: Los agentes biológicos, tales como las esporas fúngicas suspendidas en el aire, en sitios
donde se conservan colecciones científicas, pueden dar lugar al ataque y deterioro de las mismas por los
hongos.
Objetivos: El objetivo de este trabajo fue proporcionar información acerca de la calidad del aire interior en
el Herbario de Plantas Vasculares del Museo de Ciencias Naturales de La Plata, Argentina, en relación con el
contenido de propágulos fúngicos y partículas inertes, mediante el uso de dos técnicas complementarias.
Métodos: El estudio se llevó a cabo con un sistema volumétrico y dos metodologías de muestreo:
1) método no viable de evaluación directa; y 2) método viable para el cultivo de propágulos fúngicos
viables.
Resultados: A partir del sistema de recuperación directa se cuantificó un total de 4401,88 esporas/m3
con 10 morfotipos pertenecientes en su mayoría a anamorfos de Ascomycota. Asimismo se cuantificaron 32135,18 partículas inertes suspendidas por m3 . Con el uso del sistema viable se estimó un
total de 40.500 UFC/m3 aire para todos los sitios muestreados y se identificaron nueve taxa fúngicos
∗ Corresponding author.
E-mail address: malloa2001@yahoo.com.ar (A.C. Mallo).
http://dx.doi.org/10.1016/j.riam.2016.05.003
1130-1406/© 2016 Asociación Española de Micologı́a. Published by Elsevier España, S.L.U. All rights reserved.
100
A.C. Mallo et al. / Rev Iberoam Micol. 2017;34(2):99–105
que pertenecen también a formas anamórficas de Ascomycota, aunque se halló la levadura pigmentada
Rhodotorula F.C. Harrison (Basidiomycota).
Conclusiones: Ambos métodos, viable y no viable de muestreo, son necesarios para el control de la carga de
aerosoles en el Herbario de La Plata. El aire interior de esta institución parece razonablemente adecuado
para la conservación de plantas vasculares, dado el bajo índice interior/exterior, bajas concentraciones de
esporas o la ausencia de indicadores de problemas de humedad.
© 2016 Asociación Española de Micologı́a. Publicado por Elsevier España, S.L.U. Todos los derechos
reservados.
Biodeteriogens like fungi, bacteria and Actinomycetes pose
severe threat to biological collections, represent risk for human
health, and are a cause of deterioration of different stored
materials.5,14,38 Climatic and indoor environmental conditions such
as temperature, humidity and air circulation influence the microbial prevalence.41 Microbes enter the indoor atmosphere through
wind currents, and staff and visitors carry dust particles deposited
by impaction. One of the main factors which influence indoor
pollution levels is the outdoor air quality.25,29 Also, the indoor concentrations of fungal spores depend on both indoor and outdoor
sources as well as on removal processes, such as air exchange and
chemical reactions.33,36
Microorganisms can compromise the structure and function of
the materials involved in biological collections in different ways.
Fungi are an important cause of chromatic and structural alterations due to mycelial growth and pigment production, degrading
cellulose and producing acids like oxalic, fumaric, succinic and
acetic acid, which alter the normal pH of the substrate.20,21
Biological collections are of vital importance for cultural heritage worldwide since they constitute an invaluable source of
scientific information. In this sense, numerous standardized protocols are applied at present to protect and maintain biological
materials over time.47 Biodeterioration and biodegradation processes are problems that curators have to deal with nowadays.
The monitoring and control of these agents that may colonize
and degrade materials are critical measures for the safe preservation of this cultural heritage. Several studies about the prevalence
and viability of microbial propagules in the air have been carried
out.12,16,51 These studies are vital to have a detailed overview of
the environmental air quality inside biological collections16 due
to their usefulness as indicators of the conditions of conservation.
The handling of biodeteriorated specimens is another serious problem for people’s health since these microorganisms might be also
pathogenic, allergenic and/or toxinogenic.27,35,45,46
The biological material stored in herbaria are mostly collections
of dry botanic specimens, organized by a given system, classified by
a phylogenetic and alphabetic order6 and preferably stored under
controlled conditions for conservation. The specimens are mounted
on cardboard and every material must be stored in cold (−20 ◦ C)
quarantine before incorporation. Then, they are transferred to special cabinets for preservation. One of the problems of conservation
of these collections is related to the substrate, comprised by both
the plant material (each specimen) and the support (cardboard),
because both are rich in organic components for the development
of insects, fungi and bacteria, which are important biodeterioration
agents.34,51 Another problem of these collections is the phytosanitary conditions of the plant material itself. So, safety practices as
the control of environmental conditions, people movement, and
monitoring of the collections are standard measures to be taken.
The Museo de Ciencias Naturales de La Plata, Argentina (34◦ 550 S,
57◦ 570 W) (Fig. 1a and b), houses the Herbarium of Vascular Plants
(LP), which is an important institution containing significant collections of Asteraceae, Poaceae, Fabaceae and Pteridophyta (ca.
500,000 specimens). This institution is one of the largest in Latin
America, and includes type material (5000 specimens) collected
by famous botanists such as A. L. Cabrera and C. L. Spegazzini.19,26
Although the plant collection is properly preserved in special cabinets and with controlled handling of environmental conditions,
there is no available information about the indoor air quality of
this Herbarium.
Fungi have proved to be the most important biodeterioration
agents, especially on supports made of cellulose,44 which is the
main component of these plant collections. On the other hand, dust
serves as a source of nutrients to some insects and fungi, and creates
a microenvironment on collection surfaces, facilitating the absorption of moisture and promoting the proliferation of pests.13 For this
reason, the aim of this work was to characterize both the indoor air
mycobiota and the content of inert particles in the LP Herbarium,
by using viable and non-viable volumetric methodologies.
Material and methods
Study site
The LP Herbarium, with a total surface of 350 m2 and ca.
1400 m3 , has several workplaces (Fig. 1c), including areas for
research and administration, and a main room where plant specimens are adequately preserved under standard protocols and
controlled temperature and humidity conditions (Laura Iharlegui,
personal communication). Taking into account the age of the building, the place where the collections are stored (sites I and II) is in
good condition due to its regular maintenance. The working and
circulation areas (sites IV and V) do not present visible deterioration, whereas the secondary entrance (III) is the most exposed
site due to the staff movement and the direct connection with the
exterior and the exterior corridor (VI). Six representative areas of
the Herbarium were analyzed in this study: I – the main access to
the Herbarium (180 m3 ), II – the central corridor (43 m3 ), III – the
secondary entrance (27 m3 ), IV – the visiting room (41 m3 ), V – the
lobby (60 m3 ), and VI – the exterior corridor (open space) (Fig. 1c).
Sampling and data analysis
The indoor air was sampled on December 21st, 2011, at 9:30 am.
The collection room (sites I–II) has a central air conditioning system
and a dehumidifier device to control temperature and humidity.
These parameters were recorded using a HOBO U14 LCD data logger. At the moment of the sampling, the mean temperature and
mean relative humidity (RH) inside the collection site were 21.6 ◦ C
and 53.5% respectively, while those outside were 27.5 ◦ C and 55%
respectively. The study was performed during a period of recess of
activities and absence of staff and visitors to reduce the effect of
turbulence in the air.
The indoor air sampling was done on a volumetric Hirst type
®
system with a Zefon Z-Lite IAQ Air Sampling Pump by using two
procedures: a non-viable and a viable method. In the non-viable
®
method the pump was connected to the Air-O-Cell cassette for
direct microscopic evaluation of bioaerosols, whereas in the viable
method a holder with a cellulose filter was connected to the same
device to obtain a sample to culture viable fungal propagules. Both
A.C. Mallo et al. / Rev Iberoam Micol. 2017;34(2):99–105
a
101
b
VI
V
IV
Vascular plants division
I
II
Herbarium
III
c
Fig. 1. Museo de La Plata and its Herbarium: Panoramic view (a), map of the left side of the Museum (b), diagrammatic scheme of the facilities of the Herbarium (c): Sampling
sites: I – main access to the Herbarium, II – main corridor, III – secondary entrance, IV – visiting room, V – lobby, VI – exterior corridor. Maps: Arch. García Santa Cruz et al.
Laboratorio de Arquitectura y Hábitat Sustentable, Universidad Nacional de La Plata.
sampling procedures were standardized at the same air flow and
time of sampling for comparison of results. The sampling was carried out for 5 min at 1.5 m high in the center of the rooms.
Briefly, in the non-viable system the air stream is accelerated
by the suction pump at a flow of 15 l/min and impacts on a glass
slide that contains a sticky and optically clear sampling medium
which collects and holds bioaerosols and inert particles. The glass
cover slip containing the sample traces was removed from the
®
Air-O-Cell cassette and mounted on a slide, then stained with
lactophenol cotton blue and covered with a slip. These samples
were observed using an Olympus BH2 microscope at a magnification of 400× along seven transverse lines covering 22% of the total
area of the preparation. Likewise, a magnification of 1000× was
used in some cases to achieve an accurate identification. Different bioaerosols, including sexual and asexual fungal spores, were
detected and identified with reference atlases of the Kingdom Fungi
and related ones,3,18,24,28,39 as well as with a specialized electronic
database.7 Aerosol counts were then converted into elements per
m3 air following Baxter.4
Since environmental dust may be a source of nutrients and
support for the development of fungal microorganisms, the inert
particle content in the air was analyzed counting the particles by
means of the non-viable system. The criteria of the American Conference of Governmental Industrial Hygienists on Total Suspended
Particulates (TSP < 100 mm) was applied for particles of inorganic
origin like opaque particles and hyaline fibers and of human origin
like skin cell fragments.28 Particle counting and estimation of concentration were made by the same methodology as that used for
non-viable bioaerosols.
In the case of culturable-air sampling, the cassette was replaced
by a sterile holder with a GE Osmonics mixed-cellulose ester filter of 0.43 mm pore and 25 mm diameter and the resulting system
was applied to the pump for 5 min for each sample at a rate of
15 l/min. After sampling, in the laboratory, each filter was suspended in 50 ml of sterile water under aseptic conditions and the
resulting suspension was vigorously shaken (15 min at 2000 rpm).
To recover the greatest amount of fungal propagules adsorbed on
the capture surface, the filter and aliquots (1 ml) of the suspension as well as its decimal dilutions (1:10; 1:100) were spread
onto plates containing a 2% (w/v) Corn Meal Agar (CMA) medium
amended with glucose (2 g l−1 ), chloramphenicol (50 mg l−1 ) and
streptomycin (100 mg l−1 ). This medium is broadly used for recovery of fungi from several types of samples such as soil, litter and
air.8,40,42
The plates were incubated at 25 ◦ C and 63% HR in the dark until
colony development. After one week, colonies which had developed on each plate were counted (Colony Forming Units, CFU),
analyzed microscopically, and identified taxonomically, based on
cultural and morphological features.11 The CFU recovered from
each sample were then converted into volumetric units calculated
as: number of colonies x volume dilution/volume of air sampled.
Furthermore, some representative fungi were isolated in axenic
culture. Stock cultures of these isolates were kept at 4 ◦ C on 2%
(w v−1 ) malt extract-agar slants, and then deposited in the culture
collection of the Instituto Spegazzini, Universidad Nacional de La
Plata, La Plata, Argentina (LPSC).
On the data obtained from both sampling procedures, the
specific prevalence48 and species richness32 of each taxon were
estimated. The indoor/outdoor index was also estimated.31
Results
Sample analysis using the non-viable method
Ten spore morphotypes from fungal and related sources, one
fern spore type and one pollen grain type were found as biological
102
A.C. Mallo et al. / Rev Iberoam Micol. 2017;34(2):99–105
Table 1
Fungal bioaerosols identified and total concentrations estimated for the non-viable sampling in bioaerosols/m3 air, and for the viable sampling in CFU/m3 in the indoor air
of the LP Herbarium.
Spore type (non-viable system)
SI
SII
Alternaria
Arthrinium
Aspergillus/Penicillium
Ascospores
Chaetomium
Cladosporium
Dreschlera/Bipolaris
Epicoccum
Leptosphaeria type
Myxomycota
Pollen
Fern spores
Total part/m3 air
Indoor/outdoor index
SIII
SIV
SV
SVI
Total count
238
59
59
953
119
238
59
1013
178
59
1788
178
59
238
357
59
178
4411
59
59
1609
178
178
59
59
119
298
59
59
1192
0.48
Fungal taxa (viable system)
SI
Acremonium
Alternaria alternata
Cladosporium cladosporioides
Cladosporium herbarum
White yeast
Penicillium frequentans
Penicillium rubrum
Penicillium thomii
Rhodotorula sp.
Total part/m3 air
Indoor/outdoor index
SII
SIII
500
2000
59
178
536
0.21
178
0.07
SIV
2504
SV
SVI
Total count
500
1500
500
1500
500
500
1500
2500
0.63
27,500
30,000
7.50
500
0.13
2000
0.50
500
3000
4000
500
1000
500
500
2500
28,000
40,500
500
500
500
500
500
500
500
500
4000
1500
0.38
Table 2
Type and concentrations of inert particles (particles/m3 air) estimated in the indoor air of the LP Herbarium.
SI
SII
Skin cell fragments
Hyaline fibers
Opaque particles
Total
1013
1311
59
2384
8048
1669
9777
SIII
5842
4709
10,552
dispersal units in the samples analyzed (Table 1). Other dusty particles such as skin cell fragments, hyaline fibers, and opaque particles,
were also identified (Table 2). Total aerosol count converted into
elements per m3 air corresponding to all the sites sampled showed
a total of 4401.88 biological spores and 32135.18 total suspended
particles (TSP).
Regarding the spore concentration, no spores were found in
the main access (I) or in the central corridor (II) of the rooms
containing the collections. In contrast, the secondary entrance
(III) showed a concentration of around 1200 units/m3 , where
Aspergillus/Penicillium and Cladosporium spore types were prevalent. In the visiting room (IV), the total concentration was of about
180 units/m3 and the ascospore type was the most important. The
total spore concentration in the lobby (V) was 350 units/m3 and
Dreschlera/Bipolaris was the most important type. Also fern spores
and pollen grains were found. Finally, the highest concentration of
spore types was found in the exterior corridor (VI), with around
2500 units/m3 (Fig. 2) and Cladosporium being the prevalent type
(Table 1).
Regarding the results obtained in the sampling of inert particles (Table 2), skin cell fragments represented 67% of the total
concentration, hyaline fibers 24% and opaque particles 9%, being
the central corridor (II) and the secondary entrance (III) the most
affected by TSP.
Since the magnitude of the outdoor bioaerosol concentration
might also be a determining factor affecting the indoor levels, an
indoor/outdoor index was estimated for all interior sites and for
individual sporal types, using the data obtained in the exterior corridor (VI) as an outdoor reference. The indoor/outdoor index in the
SIV
SV
SVI
775
417
715
1132
5723
1192
536
7452
Total count
21,403
7631
3040
32,135
59
834
3000
2500
Spores/m3
Inert particles
2000
1500
1000
500
0
S1
S2
S3
S4
S5
S6
Sample
Fig. 2. Concentrations obtained by the non-viable method in the sampling sites
(spores/m3 air).
collection room was 0% in the main access (I) and central corridor
(II) and 46% in the secondary entrance (III). Values for the visiting room (IV) and the lobby (V) were of 7% and 21% respectively
(Table 1).
When the ratio was calculated for individual taxa (Table 1), four
sporal types were present only in the exterior corridor (VI), two
solely in the interior air of the Herbarium and four taxa both inside
and outside (Fig. 3).
A.C. Mallo et al. / Rev Iberoam Micol. 2017;34(2):99–105
9
Alternaria
Arthrinium
Chaetomium
Epicoccum
8
Vs
NVs
7
Number of taxa
Aspergillus/Penicillium
Cladosporium
Leptosphaeria type
Myxomycota
103
Outdoor
Indoor
6
5
4
3
2
Shared
1
0
S1
S2
S3
S4
S5
S6
Sample
Fig. 5. Comparison of the richness of taxa obtained by the viable and non-viable
methods in the sampling sites. V, viable method; NV, non-viable method.
Ascospore
Dreschlera/Bipolaris
Fig. 3. Typical spores found by the non-viable method, inside the Herbarium, outside the Herbarium and in both sites.
Alternaria alternata
Cladosporium cladosporioides
Penicillium thomii
Rhodotorula
Cladosporium herbarum
Penicillium frequentans
Penicillium rubrum
30 000
CFU/m3
25 000
20 000
Outdoor
15 000
Indoor
Shared
10 000
5000
Acremonium
0
S1
S2
S3
S4
S5
S6
Sample
Fig. 4. Concentrations obtained by the viable method in the sampling sites (CFU/m3
air).
Sample analysis using the viable method
Nine fungal taxa were identified as viable spores when different air samples from the LP Herbarium were applied as inoculum
source on an agar medium (Table 1). Although several decimal dilutions of up to 1:103 were spread on agar plates, only informative
data were obtained in the sampling of their own filter and the
non-diluted suspension (≤40 CFU per plate).
The total estimated count of fungal CFU/m3 air showed
40500 elements for all the sites sampled.
Regarding the total concentration, 2500 CFU/m3 air were estimated for the main access (I), where Cladosporium cladosporioides
(Fresen.) G.A de Vries, was the most important type. The central corridor (II) showed the lowest value, with 500 CFU/m3 and Alternaria
alternata (Fr.) Keissl, being the main type. The secondary entrance
(III) had the highest concentration, with 30,000 CFU/m3 , mainly
represented by Rhodotorula F.C. Harrison.
In the visiting room (IV) (2000 CFU/m3 ), Penicillium thomii
Maire, was the most important taxon being close to Penicillium
frequentans Westlingbut (Fig. 4).
When each spore type was considered, Rhodotorula sp. F.C. Harrison, C. cladosporioides (Fres.) de Vries and Cladosporium herbarum
(Pers.) Link, were the most prevalent taxa. Similarly to the data
obtained using the non-viable method, the exterior corridor (VI)
showed the highest richness of airborne fungal propagules (Fig. 5).
When the ratio was calculated for individual taxa in the viable
method (Table 1), three spore types were present in the exterior
corridor (VI), one solely in the interior air of the Herbarium and
four taxa both inside and outside (Fig. 6).
Fig. 6. Typical spores found with the viable method, inside the Herbarium, outside
the Herbarium and in both sites.
Discussion
In this study, a preliminary analysis of the indoor air quality
in six sectors of the LP Herbarium was done using a volumetric
system for both viable and non-viable samples. Although a differential richness of spores and/or other airborne elements was found,
it was dependent on each methodology applied. In this sense, the
viable method was the most efficient in the identification of fungal
taxa. While three fungal representatives were captured from sites
I and II, the non-viable one did not record any fungal spore in those
sectors. However, the apparent absence of propagules from one
sampling type cannot discard their existence in the environment,
when they are detected using another method. This fact is closely
related to the existence of specific limitations for each type of
sampling, being the morphological alteration and the viability and
physiological requirements (including nutritional ones) the critical aspects for the non-viable and viable methods respectively.43
Trunov et al.49 suggested that disaggregated spherical spores, such
as those belonging to Aspergillus and Penicillium, can be damaged
by mechanical stress during their collection and be transformed
to particulate elements that hinder their diagnosis and therefore
their identification. On the other hand, the viable method also
can fail when spore viability is affected and/or when the culture
medium used for the growth of the air samples is not suitable.
Although Aspergillus/Penicillium-type and Chaetomium-type spores
were found in the secondary entrance and/or the exterior corridor,
respectively, none of these taxa developed on an agar medium for
fungi when samples from the viable method were inoculated.
While the two methods used in this work recovered a similar number of fungal spores, the taxa were different in each case
(Figs. 3, 5 and 6). Only spores related to the genera Alternaria
Nees and Cladosporium Link were detected by both methods. However, the non-viable method, which detected a higher number of
104
A.C. Mallo et al. / Rev Iberoam Micol. 2017;34(2):99–105
taxa than the viable method, is also useful for the identification
and quantification of some ascospores, basidiospores and other
propagules that show specific difficulties to germinate.37 Both
Aspergillus-Penicillium-type and Cladosporium-type spores were the
ones that showed the highest relative concentration (23 and 40%,
respectively) in comparison to the other types identified by the
non-viable method. This method also allowed the capture and
detection of several inert particles, which might be important
in air pollution since they may be vehicle and/or substrate for
microorganisms.15,22,41 Similarly, other authors have reported a
higher number of airborne fungi using the non-viable method.1,2,7
In contrast, the viable method allows the accurate identification
of most anamorphic fungal species such as those which require
culture methods.
An indoor/outdoor ratio ≥0.63 and concentrations of
bioaerosols ≥2500 part/m3 air were found in the samples from
sites I and II. Although there are no international standards for
indoor air quality regarding bioaerosols, our results suggest a
safe air quality in the conservation areas of the LP Herbarium
according to the findings of several authors.17,31,34 Additionally,
although several taxa and/or morphological types were identified
in the indoor air of the LP Herbarium by means of both methodologies, none of the samples analyzed included representatives of
Stachybotrys Corda, Fusarium Link, Trichoderma Pers., or Aspergillus
versicolor (Vuill.) Tirab, which are generally regarded as indicators
of moisture problems.9,34 Therefore our results are compatible
with a safe conservation for cultural heritage such us Herbarium
material. However, most fungal types and taxa that we found in the
Herbarium, such as Penicillium and Cladosporium, are ubiquitous
fungal elements in indoor air.23,53 On the other hand, higher
richness and concentration of airborne elements were found in
site VI (exterior corridor), probably due to the proximity to the
exterior and/or the movement of the staff within the Museum.
Furthermore, the secondary entrance, which showed a higher
indoor/outdoor index for the culturable sample, is also amenable
to aerobiological contamination, possibly due to the fact that
it is a crossing point between the Herbarium and the outside
world, and it is frequently used to access to the collections. In
this sense, Rhodotorula sp., which was detected only in this site
(secondary entrance) using the viable method, was the element
with the highest relative concentration, in comparison to others
also identified (Table 1). Although Rhodotorula is a common environmental yeast that may be found in several contexts-including
the air-, colonizing plants, humans, and other mammals, it has
been recently recognized as an emerging pathogen in immunosuppressed patients, and the number of infections it causes
has increased along the time.50,52 The presence of this fungus
in association to a high level for an individual bioaerosol (not
detected in the exterior corridor) suggests that the main source
comes from within, for example from human origin.31 Everyday
activities in the Herbarium such as handling organic materials,
resuspension of spores as a result of cleaning activities, and the
transportation of spores in the clothes,30 plus the movement of
the air and the relative humidity, could help to explain the results
obtained.
In conclusion, both sampling methods were necessary to
monitor the bioaerosol charge in the LP Herbarium. Under the
environmental conditions in which this institution was analyzed,
the status in this indoor air seems to be rationally adequate for
the conservation of Vascular Plants due to the low indoor/outdoor
index, low concentrations of airspora and lack of taxa indicator of
moisture problems, at least in sites I and II, where the plants are
conserved. Similar results were obtained during the same season
of sampling in a neighboring repository of mummified legacy
from the Museo de La Plata.39 Therefore, although preliminary,
our results may be applied for the diagnosis and prevention of
the potential effects of airborne fungi in biodeterioration since
under certain climatic conditions such as high humidity and
precipitation, their explosion and activity may be relevant.10
These findings are important since there are scarce available
data or systematic studies on the quality of the indoor air in La Plata
Museum and the incidence of bioaerosols, including fungi, which
can potentially cause adverse health effects.39 This information
should have priority treatment to evaluate the need of measures
for the proper handling of materials, and to take any additional
action to control the conditions and safe preservation of the plant
collections of the LP Herbarium and other similar institutions.
Sources: Maps of the Museo de Ciencias Naturales and Herbarium LP by Arch. Mauro García Santa Cruz, Lic. Jimena García Santa
Cruz and Arch. Analía Gómez Laboratorio de Arquitectura y Hábitat Sustentable, Facultad de Arquitectura y Urbanismo Universidad
Nacional de La Plata.
Conflict of interest
The authors declare no conflict of interest.
Acknowledgements
The authors wish to thank architect Mauro García Santa Cruz,
lic. Jimena García Santa Cruz and architect Analía Gómez, from
the Laboratorio de Arquitectura y Hábitat Sustentable, Facultad de
Arquitectura y Urbanismo, Universidad Nacional de La Plata, Museo
de Ciencias Naturales and Herbarium LP for drawing the maps of
the Museum and Herbarium.
The authors wish to thank Dr. Pedro Balatti for his generous
advice on the development of the viable method. We are grateful
to Dr. Jorge Crisci for allowing us to carry out this study in the LP
Herbarium, to its curator, lic. Laura Iharlegui for her assistance, and
to Dr. Marta A. Morbelli for her unconditional support.
This study was supported by grants from the Consejo Nacional
de Investigaciones Científicas y Tecnológicas (CONICET), PIP 112200801-01412, PIP 112-200801-01085, PIP 112-201101-00381, PIP
112-201101-00087, PIP 112-20110100391; Foncyt: PICT 20130148, PICT 501, and Facultad de Ciencias Naturales y Museo,
Universidad Nacional de La Plata (N581). Lic. Andrea C. Mallo
belongs to the Comisión de Investigaciones Científicas de la Provincia de Buenos Aires CIC. PBA and Dr. Lorena Elíades, Dr. Daniela S.
Nitiu and Dr. Mario Saparrat are researchers from CONICET.
References
1. Adhikari A, Sen MM, Gupta-Bhattacharya S, Chanda S. Airborne viable, nonviable, and allergenic fungi in a rural agricultural area of India: a 2-year study
at five outdoor sampling stations. Sci Tot Environ. 2004;326:123–41.
2. Almaguer M, Aira MJ, Rodríguez Rajo FJ, Rojas TI. Study of airborne fungus spores
by viable and non viable methods in Havana, Cuba. Grana. 2013;52:289–98.
3. Barnet HL, Hunter BB. Illustrated genera of imperfect fungi. New York: MacMillan Publ. Co.; 1987.
4. Baxter A. On line document Air O Cell Interpretation guide. Last update on
January 2013. Environmental Analysis Association; 2006.
5. Borrego S, Guiamet P, Gómez de Saravia S, Batistini P, García M, Lavin P, et al. The
quality air at archives and the biodeterioration of photographs. Int Biodeterior
Biodegrad. 2010;64:139–45.
6. Bridson D, Forman L, editors. The herbarium handbook. Kew, UK: The Board of
Trustees of The Royal Botanic Gardens; 1992. p. 93.
7. Burge HP, Boise JR, Rutherford JA, Solomon WR. Comparative recoveries of airborne fungus spores by viable and non-viable modes of volumetric collection.
Mycopathologia. 1997;61:27–33.
8. Cabello MN, Arambarri AM. Diversity in soil fungi from undisturbed and disturbed Celtis tala and Scutiabuxifolia forests in the eastern Buenos Aires province
(Argentina). Microbiol Res. 2002;157:115–25.
9. Chapman JA. Stachybotrys chartarum (chartarum = atra = alternans) and other
problems caused by allergenic fungi. Allergy Asthma Proc. 2003;24:1–7.
10. D’Arcy N, Canales M, Spratt DA, Lai K. Healthy schools: standardization of culturing methods for seeking airborne pathogens in bio-aerosols emitted from
human sources. Aerobiologia. 2012;28:413–22.
A.C. Mallo et al. / Rev Iberoam Micol. 2017;34(2):99–105
11. Domsch KH, Gams W, Anderson T. Compendium of soil fungi. Eching, Germany:
IHW-Verlag; 1993.
12. Fazio AT, Papinutti L, Gómez BA, Parera SD, Rodríguez Romero A, Siracusano G,
et al. Fungal deterioration of a Jesuit South American polychrome wood sculpture. Int Biodeterior Biodegrad. 2010;64:694–701.
13. Florian MLE. Heritage eaters: insects and fungi in heritage collections. London:
James & James (Science Publishers) Ltd.; 1997.
14. Florian MLE. Water, heritage photographic materials and fungi. Top Photogr
Preserv. 2003;10:60–73.
15. Gallo F, Valenti P, Colaizzi P, Sclocchi MC, Pasquariello G, Scorrano M, et al.
Research on the viability of fungal spores in relation to different microclimates
and materials. In: International Conference on Conservation and Restoration of
Archive and Library Materials, vol. 1. 1996. p. 177–93.
16. Giraldo Castrillón M. Aislamiento de hongos celulolíticos causantes del biodeterioro de la Biblioteca Central de la Universidad del Valle (Cali-Colombia). Rev
Mex Micol. 2009;29:9–14.
17. Gots RE, Layton NJ. Indoor health background levels of fungi. J Occup Environ
Hyg. 2003;64:427–38.
18. Grant Smith E. Sampling and identifying allergenic pollens and molds. San Antonio, TX: Blewstone Press; 1990.
19. Gutiérrez DG, Katinas L, Torres Robles SS. Material tipo de Carlos L. Spegazzini
en el herbario del Museo de La Plata (LP), Argentina. II: Fabaceae. Darwiniana.
2002;40(1–4):77–101.
20. Hidalgo Y, Borrego S. Aislamiento y caracterización de hongos en documentos
de la Biblioteca Nacional José Martí; 2006. http://www.bnjm.cu/rev biblioteca/
bibliotecas 2006/pages/articulo6.htm
21. Hidalgo Y. Aislamiento y caracterización de hongos en documentos de la Biblioteca Nacional José Martí. Bibliotecas. An Investig. 2006;2.
22. Hsing J, Milton CH, Schwartz DK, Burge HA. Dust borne fungi in large office
buildings. Mycopathologia. 2001;154:93–106.
23. Hyvärinen A, Vahteristo M, Meklin T, Jantunen M, Nevalainen A, Moschandreas
D. Temporal and spatial variation of fungal concentrations. Indoor Air Aerosol
Sci Technol. 2001;35:688–95.
24. Käärik A, Keller J, Kiffer E, Perreau J, Reisinger O. In: Nilsson S, editor. Atlas of
airborne fungal spores in Europe. Berlin: Springer-Verlag; 1983.
25. Kanaani H, Hargreaves M, Ristovski Z, Morawska L. Deposition rates of fungal
spores in indoor environments, factors affecting them and comparison with nonbiological aerosols. Atmos Environ. 2008;42:7141–54.
26. Katinas L. Material Tipo de Carlos L. Spegazzini en el Herbario del Museo de La
Plata (LP), Argentina. III. Darwiniana. 2004;42(1–4):177–200.
27. Klich MA. Health effects of Aspergillus in food and air. Toxicol Ind Health.
2009;25(9–10):657–67.
28. Lacey ME, West JS. The air spore. Dordrecht, Netherlands: Springer; 2006.
29. Lee HS, Kang BW, Cheongs JP, Lee SK. Relationships between indoor and
outdoor air quality during the summer season in Korea. Atmos Environ.
1997;31:1689–93.
30. Lehtonen M, Reponen T. Everyday activities and variation of fungal spore concentrations in indoor air. Int Biodeterior Biodegrad. 1993;31:25–39.
31. Levetin E, Shaughnessy R, Fisher E, Ligman B, Harrison J, Brennan T. Indoor air
quality in schools: exposure to fungal allergens. Aerobiologia. 1995;11:27–34.
32. Magurran AE. Ecological diversity and its measurement. London: Croom Helm;
1988.
33. Maroni M, Seifert B, Lindvall T, editors. Indoor air quality: a comprehensive
reference book. Air quality monographs, vol. 3. Elsiever; 1995.
34. Michelsen A, Pinzari F, Barbabietola N, Piñar G. Monitoring the effects of different
conservation treatments on paper-infecting fungi. Int Biodeterior Biodegrad.
2012;30:1–9.
105
35. Muro Cacho CA, Stedford T, Banasik M, Suchecki TT, Persad AS. Mycotoxins:
mechanisms of toxicity and methods of detection for identifying exposed individuals. J Land Use Sci. 2004;19:537–45.
36. Mycology [Online]. http://www.mycology.adelaide.edu.au [accessed 17.03.14].
37. Nayar TS, Jothish PS. An assessment of the air quality in indoor and outdoor air
with reference to fungal spores and pollen grains in four working environments
in Kerala, India. Aerobiologia. 2013;29:131–52.
38. Nitiu DS, Mallo AC, Gardella Sambeth MC, Morbelli MA. Contribución a la identificación de esporas del Reino Fungi en la atmósfera de La Plata (Argentina). Bol
Soc Arg Bot. 2010;45(3–4):301–8.
39. Nitiu DS, Mallo AC, Elíades LA, Saparrat ME, Vazquez HR. Monitoreo de la carga
fúngica ambiental y de otros bioaerosoles en un depósito de restos momificados del NOA del Museo de La Plata (Argentina). Bol Soc Arg Bot. 2015;50:
427–36.
40. Pancotto VA, Sala OE, Cabello M, López NI, Robson M, Ballare CA, et al. Solar
UV-B decreases decomposition in herbaceous plant litter in Tierra del Fuego,
Argentina: potential role of an altered decomposer community. Glob Change
Biol. 2003;9:1465–74.
41. Petushkova J, Kandyba P. Aeromicrobiological studies in the Moscow cathedrals.
Aerobiologia. 1999;15:193–201.
42. Punnapayak H, Sudhadham M, Prasongsuk S, Pichayangkura S. Characterization of Aureobasidium pullulans isolated from airborne spores in Thailand. J Ind
Microbiol Biotechnol. 2003;30:89–94.
43. Pyrri I, Kapsanaki-Gotsi E. A comparative study on the airborne fungi in
Athens, Greece, by viable and non-viable sampling methods. Aerobiologia.
2007;23:3–15.
44. Saparrat MCN, Estevez JM, Troncoso MI, Arambarri AM, Balatti PA. In-vitro
depolymerization of Scutia buxifolia leaf-litter by a dominant Ascomycota Ciliochorella sp. Int Biodeterior Biodegrad. 2010;64:262–6.
45. Sequeira S, Cabrita EJ, Macedo MF. Antifungals on paper conservation: an
overview. Int Biodeterior Biodegrad. 2012;74:67–86.
46. Simmon Nobbe B, Denk U, Poöll V, Rid R, Breitenbach M. The spectrum of fungal
allergy. Int Arch Allergy Immunol. 2008;145:58–86.
47. Simmons JE, Muñóz-Saba Y, editors. Cuidado, manejo y conservación de las
colecciones biológicas, Serie Manuales de campo. Conservación Internacional.
Universidad Nacional de Colombia; 2005.
48. Soldevilla Agreda J. 2nd National Study of Pressure Ulcer Prevalence in Spain,
2005: Epidemiology and definitory wound and patient variables. Gerokomos.
2006;17:154–72.
49. Trunov M, Trakumas S, Willeke K, Grinshpun SA, Reponen T. Collections of
bioaerosols particles by impaction effect of fungal spore agglomeration and
bounce. Aerosol Sci Technol. 2001;35:617–24.
50. Tuon FF, Costa SF. Rhodotorula infection. A systematic review of 128 cases from
literature. Rev Iberoam Micol. 2008;25:135–40.
51. Velikova T, Trepova E, Rozen T. The use of biocides for the protection of library
documents: before and now. In: Méndez-Vilas A, editor. Science against microbial pathogens: communicating current research and technological advances.
2011. p. 152–3.
52. Wirth F, Goldani LZ. Epidemiology of Rhodotorula: an emerging pathogen. Interdiscip Perspect Infect Dis. 2012, http://dx.doi.org/10.1155/2012/465717. Article
ID 465717, 7.
53. Xu Z, Yao M. Analysis of culturable bacterial and fungal aerosol diversity
obtained using different samplers and culturing methods. Aerosol Sci Technol.
2011;45:1143–53.