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Airborne Fungi and Mycotoxins

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DOI: 10.1128/9781555818821.ch3.2.5

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Airborne Fungi and Mycotoxins
DE-WEI LI, ECKARDT JOHANNING, AND CHIN S. YANG

3.2.5
Fungi are a heterogeneous group of organisms including true (19–21) and are suggested as a major possible human health
fungi (Kingdom Eumycota or Fungi), lichens (a fungus and risk factor in buildings with mold problems (22, 23). In addi-
an alga in symbiotic relationship), the true slime molds (Myx- tion to mycotoxins, some MVOCs produced by actively grow-
omycetes, Kingdom Protozoa), and the water molds (Oomy- ing fungi as both primary and secondary metabolites are
cetes, Kingdom Chromista). At present the latter two are known irritants or hazardous chemicals (18, 24). They may
sometimes referred to as fungi-like organisms due to their phy- pose a health risk to building occupants and workers handling
logenetic differences from the true fungi. Frequently, they are fungal matter (5, 6, 25–27).
still called fungi, since they are traditionally studied by mycol- This chapter reviews the existing literature on airborne
ogists and covered in mycology (1). Fungi as a group inhabit a fungi with an emphasis on indoor fungal growth and contam-
wide range of niches and environments from plants, plant ination as well as the principal human health effects of expo-
debris, soils, and animals to exposed rock (some lichens), riv- sure to fungi, mycotoxins, and other by-products. It is
ers and lakes (aquatic fungi), the sea (marine fungi), the important to note that bacteria (including actinomycetes),
North Pole, and the tropics (2). They have developed mites, insects, and other microbes may also grow in a wet,
many different modes of obtaining nutrients. Fungi function damp indoor environment.
in both beneficial and detrimental ways from a human per- A wealth of literature on outdoor airborne fungi can be
spective. Some fungi, such as powdery mildews and rusts, found in reviews by Gregory (28), Flannigan et al. (29), Lacey
are obligate parasites of plants. Some fungi exist in symbiotic (30, 31), and Levetin (32, 33). It is important to note that
relationships with plant roots to form mycorrhizae or with outdoor airborne spores are often the source of indoor fungal
algae to form lichens. Some fungi are able to break down or spora. Their impact on indoor airborne fungal populations
detoxify wastes and other pollutants (3). A large number of could be immediate or delayed until they have settled and
fungi survive as saprobes recycling nutrients in ecosystems, colonized an indoor environment. Sampling of airborne
such as carbon, nitrogen, and sulfur. Unfortunately, some of and indoor fungi is not covered in this chapter. For any issues
these saprobes have been found to grow in the indoor envi- or questions related to sampling of indoor and airborne
ronments of buildings and have led to building related fungi, refer to chapter 3.2.2 in this book and several guides/
complaints and illnesses (4–7). Samson (8) estimated that books published by AIHA in the past 10 years (34–36).
100–149 species occurs indoors, while Miller and Day (9)
found approximately 270 species recovered from the dust
indoors (10). Li and Yang (11) found that over 600 species AIRBORNE FUNGAL POPULATIONS
were identified from samples collected from indoor environ- It must be emphasized that it is to the selective advantage of
ments in North America based on the information in a fungi to release, disperse, and disseminate their spores and
database of a commercial laboratory. However, the exact occasionally hyphal fragments or partial conidiophores in
number still remains unknown. Likely it will exceed 600 spe- air from one location to others. They cannot survive and
cies, as occasionally a new record is discovered from samples complete their life cycles by staying afloat in air for an indef-
collected from the indoor environment. A majority of indoor inite period of time. The dispersal of fungal spores can be
fungi are anamorphic fungi. short or long distance (37, 38). The majority disperse their
Fungi develop and release their spores into the air for dis- spores over a short distance. Therefore, when discussing fun-
persal (12). Human beings are often exposed to fungi by inha- gal contamination, identifying and locating the source of fun-
lation of airborne fungal spores, hyphal fragments, and fungal gal colonization is often of higher importance than assessing
by-products, especially in indoor environments. Certain airborne fungi data (39, 40). In addition to airborne dispersal,
fungi have been associated with asthma and respiratory con- some fungi rely on running water, insects, and animals for dis-
ditions (13–15). persal of their spores (41, 42).
Fungi produce a variety of secondary metabolites, includ- A large collection of literature on assessing indoor fun-
ing mycotoxins and some fungal volatile organic com- gal populations has been accumulated. The majority of
pounds (VOC’s), also known as microbial VOCs (MVOCs) the literature was based on air sampling data. These include
(3, 16–18). Mycotoxins are harmful to animals and humans hospitals and health care facilities (43–48), residential
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3.2.5-2 ▪ AIR

dwellings (49–55), schools (56–58), and office buildings (59, constant temperature in a sealed enclosure until the water
60). The focus of hospital sampling was often on Aspergillus in the sample reaches equilibrium with the water vapor in
species, including A. fumigatus, an opportunistic human the enclosure, then aw = ERH/100. Another expression is:
pathogen (43). General fungal populations were identified
in nonhospital sampling. vapor pressure of water in substrate
aw ¼
A comprehensive assessment of fungal contamination in vapor pressure of pure water
the indoor environment should include consideration of fac-
tors such as outdoor air, air conditioning, heating and venti- A detailed discussion of water activity of fungi in food and
lation systems, ventilation mode, heating, occupant density, materials is presented by Gravesen et al. (4), Hocking and
ventilation rate, moisture (including water damage, high Miscamble (66), Li and Yang (40), Smith and Onion (67),
relative humidity [RH] in the air, and dampness), mainte- Troller and Scott (68), Adan et al. (69), and Huinink and
nance, on-site inspection, air sampling, surface and source Adan (70).
sampling, sample analysis, risk analysis, and finally remedial Many common indoor fungi are hydrophilic and require
actions (49, 61). Unfortunately, the majority of investigations aw near 1 for growth. Some xerophilic fungi, however, have
fail to follow the approach of comprehensive assessment, optimal water activity ranging from 0.65 to 0.90 (4). Both
often due to insufficient strategy, labor, time, and funds as well mesophilic and xerophilic fungi can be found in the indoor
as understanding of environments and ecosystems indoors. environment. Table 1 lists some hydrophilic, mesophilic,
An alternate approach to identifying indoor fungal contami- and xerophilic fungi and their minimum aw. It should be
nation is to focus on inspection and surface/source sampling. noted that a very short period of peak humidity, even below
saturation RH, may lead to fungal growth. Adan et al. (69)
showed that it took only 69 h for Penicillium chrysogenum
Factors Affecting Airborne Fungal Populations Thom to develop from spore germination to sporulation
Three important factors that directly affect airborne fungal and 73 h to develop mycelial mass on pure gypsum at 21°C
populations are the availability of food/substrates and free and 97% RH observed with cryo-SEM.
water for fungal growth and the methods of spore dispersal.
Other physical, chemical, and biological parameters affecting Fungal Spore Discharge Mechanisms
fungal growth, and subsequent airborne fungal populations, Fungal spores are released by two basic mechanisms: (1)
can be found in recent references (2, 3, 8, 41). active spore discharge and (2) passive spore release. Concen-
trations of certain airborne fungal spores have been known to
Substrates (Including Water Activity) peak during certain hours of the day or night. This periodicity
Fungi are achlorophyllous and heterotrophic, take up is related to spore discharge mechanisms and environmental
nutrients by absorption from substrates, and require simple factors in nature (28, 30, 42). Details of these two mecha-
sugars, carbohydrates, and other organics, such as vitamins, nisms and environmental factors affecting spore release are
amino acids, and essential mineral elements to survive (3). presented by Lacey (30) and Levetin (33).
In the natural environment, fungi have developed a number Fungi with active spore discharge include such common
of ways to obtain these nutrients (3), such as necrotrophic, airborne fungi as Sporobolomyces, Epicoccum, Nigrospora, and
symbiotic, and saprotrophic relationships. some smut-like yeasts. Many ascospores and basidiospores
Humans share food, living space, environments, and also have active discharge mechanisms (33). Sporobolomyces
resources with fungi. We utilize fungi to produce bread; man- and some basidiospores are usually most abundant at night
tou (steamed bun); cheese; edible mushrooms; alcoholic bev- or in the predawn hours. Their spore release requires the
erages; and useful by-products, such as antibiotics, enzymes, absorption of moisture to build-up release pressure or forming
and organic acids (60). Some fungi cause food spoilage or a droplet at the hilar area. Dry spore fungi, such as Aspergillus,
make food toxic to humans. Botrytis cinerea is a well-known Penicillium, and Cladosporium, are often hydrophobic. They
pathogenic fungus causing gray mold disease on grapevines, become airborne by passive force, such as air movement or
strawberries, and many other fruits and produce. Species of rain droplets (71, 72). Cladosporium usually dominates the air-
Penicillium and Aspergillus often cause spoilage in foodstuffs borne spore population during the day. Its spores stay airborne
and make them inconsumable to animals and humans. On owing to the buoyancy of warmer air.
the other hand, Penicillium camemberti and P. roqueforti are Some fungi, such as puffballs (Calvatia, Lycoperdon, Sclero-
used in cheese production. Fungi are also known to cause derma) of basidiomycetes, are able to produce a huge number
wood stains, wood decay, biodeterioration, and biodegrada- of spores and release their spores in “spore clouds or puffs”
tion of polymers, carpet, plaster, drywalls, wallpaper, paints when affected by raindrops, humans, or small animals (33,
and organic coatings, fuels and lubricants, leather, fabric 71–73). The spore clouds may persist for a period of time until
products, paper, and wood products (4, 62–65). These refer- air mixing and dilution disperse them. Results of air sampling
ences underscore the very likelihood that fungi can and will can be greatly affected by whether the spore cloud has dis-
grow in artificial environments. Consequently, controlling persed (31).
nutrient sources to limit fungal proliferation/growth is practi- Many fungi that are frequently detected indoors and out-
cally impossible. doors produce spores in a slimy mass. These include such
One of the critical factors affecting indoor fungal growth is common indoor contaminants as Acremonium (although
water. There are a number of ways to measure water availabil- some species of Acremonium produce dry spores), Aureobasi-
ity in materials. Water or moisture content of a material is dium, Fusarium, Phoma, Stachybotrys, Trichoderma, and yeasts.
expressed as a percentage of the oven dry weight (65). How- Slimy spores may be released into the air when they become
ever, water content does not suggest the actual availability of dry, disturbed, or attached to other particles. Their dissemina-
free water in the material to fungi. A better measurement of tion is often assisted by insects, mites, small animals, or water
water availability to fungi is water activity. Water activity is (42). Because slimy spores do not become airborne easily,
numerically equal to equilibrium relative humidity (ERH) their detection indoors should be considered significant.
expressed as a decimal. If a sample of substrate is held at Any detection of Stachybotrys in air samples taken indoors

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TABLE 1 Selected fungi and their aw b


aw T (°C) aw T (°C)
Absidia corymbifera 0.88 25 Paecilomyces variotii 0.84 25
Alternaria citri 0.84 25 Penicillium brevicompactum 0.81 23
Aspergillus candidus 0.75 25 P. chrysogenum 0.79 25
A. flavus 0.78 33 P. citrinum 0.80 25
A. fumigatus 0.82 25 P. corylophilum 0.80 25
A. niger 0.77 35 P. expansum 0.83 23
A. ochraceus 0.77 25 P. frequentans 0.81 23
A. restrictus 0.75 25 P. griseofulvum 0.81 23
A. sydowii 0.78 25 P. spinulosum 0.80 25
A. terreus 0.78 37 Phoma herbarum 0.93 25
A. versicolor 0.78 37 Rhizopus microsporus 0.90 25
A. wentii 0.84 25 R. stolonifer 0.84 25
Cladosporium cladosporioides 0.84 25 R. oryzae 0.88 25
Cladosporium sphaerospermum 0.84 25 Scopulariopsis brevicaulis 0.90 NA
Eurotiuma amstelodami 0.70 25 Sistotrema brinkmannii 0.97 25
Eurotium chevalieri 0.71 33 Stachybotrys chartarum 0.93 25
Emericellaa nidulans 0.78 37 Syncephalastrum racemosum 0.84 25
Fusarium moniliforme 0.89 25 Trichoderma harzianum 0.91 25
Geomyces pannorum 0.89 25 Wallemia sebi 0.70 25
Mucor circinelloides 0.90 25 Ulocladium chartarum 0.89 25
M. plumbeus 0.93 25 U. consortiale 0.89 25
a
Both genera are teleomorphs of Aspergillus.
b
NA, not available. Sources: 40, 65–67; For calculation of aw, see text.

should trigger further investigation and search for the con- and they are often missed or misidentified. A real-time
firmed presence of this fungus. PCR test has been developed for the detection of dry-rot
fungi, Meruliporia incrassata and Serpula lacrymans (83).
Airborne Fungal Populations Ascospores have been identified from air samples using
Fungi as a group produce a number of different spores, both spore traps (84). Many Ascomycetes, however, do not pro-
sexual and asexual. Many types of spores are capable of duce ascomata and ascospores in culture. This makes it diffi-
becoming airborne. Some spores, such as chlamydospores cult to determine the frequency of occurrence of ascospores in
(asexual) and zygospores (sexual), are not designed to be dis- air. Some ascomycetes, such as Ascotricha, Chaetomium,
persed by air transmission, and there has been no report of Emericella, and Eurotium (both Emericella and Eurotium are
recovering these spores in air or they are rarely observed in teleomorphs of some Aspergillus species) are commonly
the air, such as spores of Glomus species (Klironomos, unpub- formed in cultures. Ascomata of species of Ascotricha, Peziza,
lished data). Hypogeous fungi (including such well-known and Pyronema are frequently observed or reported from indoor
fungi as truffles) produce subterranean sporocarps and dis- environments (11, 85). This demonstrates that ascospores
perse their spores through different mechanisms. However, may become airborne. Ascospores are suspected to be aller-
both sexual and asexual spores of four major groups (Myxo- genic. Four species of Chaetomium were listed as licensed
mycetes, zygomycetous fungi, Ascomycetes including ana- by the FDA for commercial production as allergens (84).
morphic fungi [formerly classified as Deuteromycetes], and Rivera-Mariani et al. (86) demonstrated that out of 33 aller-
Basidiomcetes) have been isolated and reported from air. gic rhinitis and asthmatic patients in Puerto Rico, 31 reacted
Spores of Ascomycetes and Basidiomycetes have fre- to ascospores, 29 to basidiospores, 19 to hyphae/fungal frag-
quently been recovered from air samples using spore trap sam- ments, and 12 to mitospores. They stated that sensitization
plers. Levetin (41) reported 18 genera of basidiospores from to airborne spores of basidiomycetes, ascomycetes, and fungal
the atmosphere in Tulsa, Oklahoma. Furthermore, many spe- fragments seem to be more prevalent than to similar numbers
cies of basidiospores have been demonstrated to be allergenic of conidia in patients with active allergies, suggesting a possi-
(74–80). The importance of basidiomycetes in the indoor ble role in exacerbations of respiratory allergies in tropical
environment depends on the building construction. Wood- environments. Otherwise, little documentation is available
inhabiting basidiomycetes, such as polypores, are associated on the allergenicity of ascospores.
with wood decay and may be found in buildings constructed The majority of airborne fungi collected on samplers and
of wood. In fact, wood decay caused by polypores is a signifi- grown on agar media are anamorphic fungi and zygomycetous
cant problem in the United Kingdom (81) and in the United fungi. Most anamorphic fungi are asexual states of Ascomy-
States (65). A number of Coprinus species and several other cota and a minority Basidiomycota. Asexual spores of ana-
mushrooms were found growing in buildings with water dam- morphic fungi are called conidia, and of zygomycetous
age (82). Unfortunately, identification of airborne basidio- fungi, sporangiospores. Many of these spores are known aller-
mycetes collected using culture techniques can be difficult gens. Some of them have been prepared into allergen extracts

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3.2.5-4 ▪ AIR

and approved by the FDA for medical uses (84). Common Mucor, Paecilomyces, and yeasts were the five most common
anamorphic fungi found in air include Alternaria, Aspergillus, fungi recovered in culture at 37°C. The agreements and dis-
Cladosporium, Epicoccum, and Penicillium. Mucor, Rhizopus, agreements in the findings may be attributed to differences
and Syncephalastrum, all zygomycetous fungi, are also fre- in sampling techniques, isolation media used, incubation
quently isolated from air. temperatures, and geographical areas (42, 88).
Another group capable of producing and releasing spores Shelton et al. (91) evaluated 12,026 fungal air samples
into the air is the Myxogastria (formerly Myxomycota) or (9,619 indoor samples and 2,407 outdoor samples), collected
the true slime molds. Slime molds, as a group, are polyphy- from 1,717 buildings located across the United States using
letic. They are considered to have similarities to both true Andersen N6 single-stage samplers. Ninety-nine percent of
fungi and animals (1) and have been placed in the Kingdom the samples were collected with rose bengal agar and the
Protista (33, 87). Spores of slime molds have been docu- other 1% with malt extract agar. The culturable airborne fun-
mented from air samples (28, 87). Occasionally, species of gal concentrations in indoor air were lower than those in out-
Stemonitis have been found growing in environments such door air. However, Stachybotrys chartarum was identified in
as a cellar, basement, and window sill underneath a leaking the air in 6% of the buildings studied and 1% of the outdoor
window air conditioning unit in buildings with water issues samples. The fungal levels were highest in the fall and summer
( personal observation). Allergic reactions to extracts of slime and lowest in the winter and spring. Geographically, the high-
molds have been reported. Giannini et al. (77) reported that est fungal levels were found in the southwest, far west, and
15.4% of patients tested showed positive skin reactions to southeast. Because different fungal isolation media are known
extracts of Fuligo septica, Lycogala epidendrum, and Stemonitis to have different selective effects, the combined use of the
ferruginea. Benaim-Pinto (74) and Santilli et al. (79) found data derived from two different media is inappropriate. The
that patients yielded positive skin responses to spore extracts reliability of fungal identification, data, and sampling quality
of Fuligo septica. control of such a large project must be scrutinized before the
Outdoor airborne fungal populations may directly or indi- results and conclusions are fully accepted.
rectly affect the indoor populations as the pathways of infiltra- In a study of 50 single-family detached homes built since
tion are often suspected to be from leaks and cracks or through 1945 with less than 0.18 m2 (or 2 ft2) of known water damage,
doors, windows, and air intake systems. Therefore, it is not and located within a central city census tract in the metropol-
surprising that common outdoor fungal taxa are often the itan Atlanta city (DeKalb and Fulton Counties) of Georgia,
predominant fungal types detected indoors (42, 42, 88). In air and dust samples were collected for assessment to establish
a review of the literature, some agreements and disagreements a baseline of “normal and typical” types and concentrations of
exist on the predominant fungi identified indoors. Yang et al. fungi in urban homes (92). The homes were predetermined
(88), based on the culture of over 2,000 Andersen samples not to have noteworthy moisture problems or indoor fungal
collected outdoors and in nonresidential buildings in the growth. The homes were sampled twice (summer and winter)
United States, found that Cladosporium, Penicillium, Aspergil- within a calendar year. Air samples were collected with a Spi-
lus, Basidiomycetes, and Alternaria were the top five fungal ral Air System at 180 lpm onto MEA plates. Positive-hole cor-
taxa found indoors as well as outdoors in frequency of occur- rection was applied. Dust samples were sieved and inoculated
rence. All five fungal taxa were detected in less than 40% of by the “direct plating” method onto MEA and DG18 media.
indoor samples. However, Cladosporium was found in over Cladosporium cladosporioides, Cladosporium sphaerospermum,
80% of outdoor samples, and Penicillium was detected in and Cladosporium spp. were the top three fungal species and
58%. These suggest that both Cladosporium and Penicillium group in both indoor and outdoor air samples. The findings
are common in outdoor air. The results were somewhat in included that rankings by prevalence and abundance of the
agreement with those reported by Strachan et al. (52) from types of airborne and dustborne fungal spores did not differ
British homes and by Womble et al. (89) in 86 office build- from winter to summer, nor did the rankings differ when air
ings in the continental United States. Strachan et al. (52) samples taken indoors were compared with those taken out-
found that Penicillium, Cladosporium, and Basidiomycetes doors. Water indicator fungi (such as Chaetomium, Stachybo-
(including Sistotrema brinkmanii) were the common types of trys, and Ulocladium) were essentially absent from both air and
mold isolated as well as the predominant mold concentrations dust samples.
measured. Womble et al. (90) found, in rank order, nonspor-
ulating fungi, Cladosporium, Penicillium, yeast, and Aspergillus Indoor Sources of Fungi
were the most common fungi indoors. Using a number of The detection of airborne fungi does not necessarily suggest
different types of samplers and sampling media, VerHoeff growth and reproduction of fungi indoors. However, it is
et al. (53) found that species of Cladosporium, Penicillium, believed that actively growing fungi in the indoor environ-
and Aspergillus (including teleomorph Eurotium) were com- ment are the primary cause of adverse health effects due to
mon in homes in the Netherlands. In a survey of 10 elemen- exposure to indoor fungal allergens, mycotoxins, and fungal
tary schools in southern California using an Andersen MVOCs. It is therefore important to identify and detect infes-
sampler, Dungy et al. (56) found that Cladosporium, Alterna- tation sites of fungi indoors. Due to the limitation of correct
ria, Penicillium, sterile mycelia, and Epicoccum were the top identification of fungi, some of the health effects of indoor
five fungal groups isolated indoors. The predominant fungi fungi are arguably considered not necessarily species-specific,
detected outdoors were slightly different in that Aureobasi- nevertheless, correct identification of fungi may be important
dium was more frequently encountered than Epicoccum. How- for practical and research issues.
ever, using a Roto Rod sampler, they found that spores of Many common indoor fungi are strong biodeteriorating
Alternaria, rust fungi, Cladosporium, Epicoccum, and smut agents and have been reported from various building materi-
fungi were predominant both indoors and outdoors. Through als and systems. Raper and Fennell in their classic publication
further comparison of airborne fungal populations in the same (93) reported various Aspergilli from building materials, such
study, the authors found that the top seven fungal types were as wallpaper and paper products, textiles, jute, insulation
identical at schools and at homes. In a hospital sampl- materials, and fabrics. Two species of Aspergillus were isolated
ing, Solomon et al. (45) found Aspergillus fumigatus, A. niger, from and found to grow on glass (94). Many species of the

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genus Penicillium, commonly detected in indoor air sampling, actinomycetes. They sampled surfaces of filters and fans with
were frequently referred to as food spoilage and biodeteriorat- RODAC contact plates and water from humidifiers. The
ing agents (4, 64, 85, 93). Penicillium chrysogenum is reported serial dilution method was used for analyzing humidifier
to be the most common fungus indoors (95). However, Scott water. A wide variety of fungi were identified. However,
et al. (95) found that P. chrysogenum is rare in outdoor air. some of the identified fungi were likely spore contaminants
P. chrysogenum was further studied by Houbraken et al. (96) rather than the result of fungal growth. Kemp et al. (104)
using partial β-tubulin, calmodulin, and RPB2 data sets. Hou- studied fungal growth on filters of the HVAC system and
braken et al. (96) found that Fleming’s penicillin-producing reported isolation of Aspergillus niger, A. fumigatus, Alternaria,
strain is not Penicillium chrysogenum but P. rubens. It is, there- Cladosporium, Mucor sp., Aspergillus sp., and Penicillium sp.
fore, P. rubens that is one of most common fungi in indoor However, they could only confirm growth of Aspergillus sp.,
environments, not P. chrysogenum s. str. Gravensen et al. Cladosporium sp., and Penicillium sp. when filters were directly
(4) included a list of 13 fungal species as important molds examined under the microscope. Buttner et al. (105) reported
in damp buildings. Samson et al. (7, 74) described some com- a controlled study using three air duct materials (i.e., gal-
mon fungal species from indoor environments. vanized metal, rigid fibrous glass ductboard, and fiberglass
Morgan-Jones and Jacobsen (63) studied moldy carpets, duct liner) and Penicillium chrysogenum spores. They found
plasterboard, and wallpaper from three hotels in Florida and that fungal growth might occur on a variety of duct materials,
Georgia. Their brief literature review suggested that many including bare metal, provided soiling and moisture were
fungi had been reported to cause biodeterioration of paper, present. The results showed that contaminated air ducts
textiles, and plaster. The genera of fungi most often identified might expose building occupants to high concentrations of
were the ascomycete genus Chaetomium; dematiaceous spores dispersed from fungal colonies growing on duct materi-
hyphomycete genera Alternaria, Cladosporium, Stachybotrys, als during normal operation of the system. Yang (102) exam-
and Ulocladium; the moniliaceous hyphomycete genera Acre- ined and cultured 1,200 fiberglass insulation liner samples
monium, Aspergillus, and Penicillium; and the pycnidial genus from HVAC systems in the United States and found fungal
Phoma. In the study, 14 species, including 2 new species of colonization and growth in approximately 50% of the samples
Cladosporium, in 11 genera were isolated and identified. In a studied. Fungal taxa were differentiated based on water and
study of toxicity of moldy building materials, Johanning humidity conditions. Species of Cladosporium and Penicillium
et al. (97) not only detected cytotoxicity of the materials to were primarily from areas with high RH, whereas species of
cell cultures but also identified satratoxin H and spirolac- Acremonium, Aureobasidium, Exophiala, Fusarium, Paecilomy-
tone/lactams and several groups of fungi. The fungi were iso- ces, Phoma, Rhodotorula, Sporobolomyces, and yeasts were sus-
lated from gypsum wallboards and other building materials. pected in areas subjected to frequent wetting. Cladosporium
The fungi identified included those described by Morgan- cladosporioides, C. herbarum, and C. sphaerospermum were
Jones and Jacobsen and additional species of Aspergillus, Pae- the primary species identified.
cilomyces, and Trichoderma. All fungi found to colonize building materials are saprobic
Li and Yang (11) described seven new records or note- and biodeteriorating agents. Some fungi, such as species of
worthy fungi isolated from indoor environments. The Chaetomium, Aspergillus, Stachybotrys, and Trichoderma, are
seven species are Ascotricha chartarum, A. erinacea, Memno- known to be capable of degrading cellulose fibers. Although
niella echinata, Sporoschisma saccardoi, Stachybotrys microspora, at least 270 fungal species have been reported from indoor
S. nephrospora, and Zygosporium masonii. All species were environments in the literature (10), it is very likely that any
recovered from water-damaged building materials, such as saprobic, biodeteriogenic, or cellulolytic fungi can potentially
drywall, wallpaper, or wood. Four species were reported for grow indoors if opportunity arises (4, 39).
the first time from the United States. Li et al. (98) described Although species of anamorphic fungi are commonly
a new species from indoor environments in the United States detected in moldy building materials, Ascotricha chartarum,
and Canada. Corda (99) described Stachybotrys chartarum as A. erinacea, Chaetomium species, Peziza spp., and Pyronema
S. atra from wallpaper of a residence in Prague in 1837. domesticum of the Ascomycetes are occasionally found on
In the past several years, mass sequencing techniques damp materials in buildings (11, 39, 79). The asexual state
were used to study indoor fungi from dust samples collected of basidiomycetes, such as species of Cryptococcus, Rhodotor-
around the world in several studies (100). Amend et al. ula, and Sporobolomyces, are also common in indoor envi-
(100) found that fungal diversity in the temperate region is ronments (39) and Wallemia sebi was often reported from
greater than that in the tropical region and building function buildings with dampness or water damage problems. In addi-
does not have significant effect on indoor fungal composi- tion, Yang (unpublished data) has seen and identified fruiting
tion, despite differences between architecture and materials structures of a slime mold, Stemonitis sp., and basidiomata
of some buildings in close vicinity based on 72 dust samples (fruiting bodies of basidiomycetes) of Coprinus spp., Pleurotus,
collected from six continents. However, Andersen et al. and Poria from various building materials, from ceiling tiles to
(101) showed that indoor mycota are associated with differ- wood products. Samson et al. (85) reported that Sistotrema
ent building materials: (a) Acremonium spp., Penicillium chrys- brinkmanii, a wood decay fungus, is commonly isolated from
ogenum, Stachybotrys spp., Ulocladium spp. and gypsum and wet, decaying window and door joinery. Mycelia and hyphae
wallpaper; (b) Arthrinium phaeospermum, Aureobasidium pul- with clamp connections, indicating basidiomycetes, are fre-
lulans, Cladosporium herbarum, Trichoderma spp., yeasts and quently detected colonizing water-damaged wood structures.
woods and plywood; and (c) Aspergillus fumigatus, Aspergillus Not surprisingly, wood-inhabiting basidiomycetes are often
melleus, Aspergillus niger, Aspergillus ochraceus, Chaetomium wood decay fungi. The wood decay, dry rot fungus Serpula
spp., Mucor racemosus, Mucor spinosus, and concrete and lacrymans is the most common indoor basidiomycete in
other floor-related materials in Denmark and Greenland. central Europe, while Meruliporia incrassata pendant to S.
In addition to building materials, fungi have been known lacrymans also received considerable attention in North
to grow in the heating, ventilating, and air-condition- America (106). Specimens of S. lacrymans from California
ing system (HVAC) (54, 102). Heinemann et al. (103) collected in nature were ascribed to var. shastensis while all
studied contamination of fungi, bacteria, and thermophilic other collections, mainly from buildings in Europe, were

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ascribed to var. lacrymans (107). Meruliporia incrassata TABLE 2 Complaints and symptoms reported by patients with
(syn. Poria incrassata) and Serpula lacrymans were detected exposure to excessive fungal growth
in building wood samples collected from the United States
by real-time PCR (83). Approximately 80 species of wood Headaches
decay fungi have been found in buildings in northern Ger- Runny nose or nasal congestion
many (82, 106). Twenty-nine species or genera of basidiomy- Burning sensation and watery eyes
cetes were identified from 3,434 decay fungi occurrences in Sore throat and hoarseness
Norwegian houses from 2001 to 2003 (108). One hundred
Sneezing or irritant-dry cough, chest tightness and burning chest
fifty-two species of wood decay fungi were reported from
sensation, shortness of breath, wheezing
wood products in the United States (109). It should be
pointed out that these fungi were not isolated from an indoor Unusual nosebleeds and coughing up blood (rare)
environment. The exact number of wood decay fungi from Skin and mucous membrane irritation (occasionally hair loss)
indoor environments in the United States remains unknown. Dizziness, concentration and memory problems; cognitive
dysfunction
Severe fatigue and exhaustion ( physical and/or mental);
FUNGI AND HUMAN HEALTH
Nausea, (vomiting) and gastrointestinal problems (loose stools,
Fungi are known in veterinary and human medicine to be a
stomachaches)
source of infections, allergies, and irritant-toxic health reac-
tions primarily with symptoms and disorders of the skin, Feverish feeling
mucous membranes, or internal organs. Current knowledge Joint and muscle ache (flu-like reaction)
and key concepts are summarized and discussed below (57,
110, 111). Expert reviews of reported health problems associ-
ated with building dampness and biological agents, such as
reactions are common in individuals with atypical fungal
the Institute of Medicine (IOM) (2004) or the WHO-EU
indoor exposure. Typical health complaints of patients living
(2009) and others concluded, based on their reviews mainly
in indoor environments with excessive fungal exposure are
of the English-language literature and epidemiological stud-
listed in Table 2.
ies, that dampness-related fungi are associated with allergies,
The medical conditions and illnesses associated with fun-
respiratory symptoms or diseases, such as asthma, and changes
gal indoor exposure include a spectrum of infectious, skin and
of the immunological system (112–116). In addition, there
respiratory disorders, allergy, and irritant/toxic health reac-
are several clinical studies and case reports of adverse health
tions (112, 114, 125, 134). Most of the reported adverse
reactions that include primarily nonallergic adverse effects
health reactions are normally of short duration and reversible
to the lungs (bleeding in infants; allergic alveolitis), neuro-
after the exposure has been stopped or controlled. In some
logical system (headaches and cognitive dysfunction), endo-
cases, the adverse health consequences can be more serious
crine and reproductive organs (thyroid hormonal changes
or may be irreversible, requiring symptomatic treatment and
and menstrual disorders in women), and rheumatological dis-
careful avoidance of microbial triggers (134). Medical condi-
orders ( joint pain), and an increased risk of cancer has been
tions are listed in Table 3 and described later in more detail.
also explored. Some of the fungi (maybe in combination with
bacteria) produce chemicals that are known genotoxins and
carcinogens (117–123). However, these are findings that Infections
are difficult to document and validate in epidemiological or Infections caused by fungi are called mycoses. They are cate-
experimental studies and have therefore been considered gorized as endemic mycoses and opportunistic mycoses.
debatable by some. Further evidence needs to be obtained. Opportunistic fungal pathogens have a great public health
Fungi and their by-products, such as (1–3)-ß-D-glucan, importance, especially in persons with HIV, with organ fail-
mycotoxins, and MVOCs, have been implicated in adverse ure, and or receiving organ transplants (135, 136). Endemic
health reactions and diseases (7,124–127). Americans spend mycoses are related to the geographical distribution of certain
up to 90% of their time indoors, where contaminants often fungal pathogens. These types of infection are caused by the
are at higher levels than they are in the ambient air. It is inhalation of airborne spores or conidia found in certain
not uncommon that exposure duration and concentrations regions where there is a higher frequency of such fungi
of atypical fungi (fungi associated with excessive dampness) because of unique soil and plant/flora conditions (30, 111,
are greater indoors than outdoors because of occupant life- 137). Table 4 lists several important fungi and the infections
styles, building conditions, and materials that lead to fungal that occur through air transmission, the diseases they cause,
growth and accumulation. Additionally, exposure to biologi- and clinical manifestations.
cal contaminants of all kinds, but particularly molds and bac- Opportunistic infections are secondary complications that
teria, can be high when buildings have moisture problems or occur in patients with an altered or weakened immune sys-
water damage (128). It is estimated that more than 1one third tem. Patients at risk for fungal infections usually have major
of buildings in the United States and Western Europe have systemic diseases or health suppressed conditions such as
severe moisture problems that result in significant fungal complicated diabetes mellitus, cancer, HIV/AIDS, severe
contamination in indoor environments (129–131). Exposure liver or kidney diseases, organ transplantation, and burn
to high levels of indoor dampness and mold has been associ- injury or may be on immune-suppressive medication treat-
ated with upper and lower respiratory symptoms, including ment. An endemic outbreak of fungal meningitis, spinal
nasal and sinus irritation, congestion and inflammation, infections, and other serious health complications in 2012
sore throats and chest burning sensation, cough, wheeze, appeared to be caused by injecting directly into the spinal
chest tightness, and exertional dyspnea in people, according fluid of patients a steroid medication for pain control, meth-
to several large epidemiological studies cited by the IOM, ylprednisolone acetate, from a compounding pharmacy
WHO, and others (112, 113, 116, 132, 133). Clinical case that reportedly was contaminated by Exserohilum rostratum,
studies and research have shown that nonallergic health Aspergillus fumigatus, Stachybotrys chartarum, Cladosporium,

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TABLE 3 Medical conditions associated with indoor fungal exposure


Organ system Clinical effect Exposure
Upper airways: Nose, sinuses, and Rhinitis, sinusitis, laryngitis Fungi, allergens, irritants,
throat MVOCs, particles
Lower airways: lung with bronchial Bronchitis, asthma, bronchiolitis, asthma, Fungi, allergens, fungal
system and alveoli allergic bronchopulmonary aspergillosis, by-products. Fine, ultra-fine
allergic extrinsic alveolitis (a.k.a. particles
hypersensitivity pneumonitis); toxic
alveolitis and pneumonitis
Combined upper and lower airway Aspergillosis Fungal rhinosinusitis Fungi, particles
Skin and mucous membrane Urticaria, dermatitis (allergic vs. irritant), Fungal irritants, allergens,
conjunctivitis
Other organs: central nervous Overlapping diagnoses, differential Fungi, organic dusts, microbial
system, immune system, liver, diagnoses (exclusion of unrelated by-products
kidney, endocrine system conditions)

and several other fungi (139). The outbreak was associated such as chronic sinusitis and asthma, suggesting a combined
with more than 39 deaths out of 620 cases in multiple states effect of infections and inflammation (146–149).
by mid-December 2012 (140). These iatrogenic fungal The prevention, diagnosis, and therapy of opportunistic
infections were likely the result of contamination during infections may be difficult for those who are not well trained
the medication production process in an unhygienic indoor and experienced in this field. Early recognition, preventive
environment. These infections are not contagious, and the building engineering, hygiene, and public health interven-
fungi are not considered obligatory pathogens. Secondary tion can reduce the incidence of mycosis, especially the insti-
fungal infections and medical complications related to air- tutional or iatrogenic acquisition in care facilities.
borne fungal contamination in hospitals and transplant units
have been reported. Immunocompromised patients may be at Allergy and Respiratory Diseases
an increased risk for opportunistic infections if pathogenic Fungi are known to cause an immune pathology with an exag-
fungi become airborne and are significantly elevated in gerated or inappropriate immune response, called hypersensi-
indoor air surveys. Among the fungi of concern are Aspergillus tivity reaction or common allergy (114). The important types
spp., such as A. fumigatus, A. flavus, and A. niger. Soil, bird of the allergic immune reactions according to the Coomb-
and bat droppings, water-damaged materials, or organic-rich Gell classification are listed in Table 5, but in the clinical
substrates in buildings may be a reservoir for these fungi context these often occur with overlapping presentation.
(39, 57, 141). Other clinically important fungal infections The fungal spore is a known cause of allergic diseases (150–
are candidiasis with local mucocutaneous or disseminated 152) and was identified as one of the major indoor allergens
systemic organ manifestations and skin mycoses, such as (114, 153). However, there are still significant methodologi-
dermatophytoses, keratomycosis, tinea nigra, piedra, and cal problems in the production of reliable allergen extract
malassezia-caused dermatitis. Invasive fungal diseases of the from fungi compared to cats, dust mites, and other better char-
paranasal sinuses may also be associated with allergic sinusitis acterized allergens. Extracts that were available often corre-
in atopic patients (142). Aspergillus species are often sponded poorly with the fungi frequentlyfound in indoor
involved. Noninvasive forms may colonize preexisting body surveys (154). Many extracts of common indoor fungi are
cavities and may be asymptomatic as long as some immuno- not available for clinical allergy testing. Several recent epide-
logical resistance can be maintained. Chronic rhinosinusitis miological studies have shown that long-duration indoor
with eosinophilic inflammation of the airways has been exposure to certain fungi can result in hypersensitivity reac-
linked to dampness-related fungi from indoor environments tion and chronic diseases. Mold levels and fungi comparable
and may also be related to the development of asthma to outside background levels and types are usually well toler-
(143–145). The beneficial use of antifungals (i.e., itracona- ated by most people. Normal or “typical” indoor molds may
zole) has been observed in treatment of respiratory conditions vary depending on climate variations and geographical

TABLE 4 Diseases transmitted by airborne fungi and affected tissuesa


Fungi Disease Affected organ/tissues
Histoplasma capsulatum Histoplasmosis Lung, eye (skin and bone)
Cryptococcus neoformans Cryptococcosis Lung, central nervous system, meninges, skin, and viscera
Coccidioides immitis Coccidioidomycosis Lung, multi-organ dissemination (skin, bone, meninges, joints)
Blastomyces dermatitidis Blastomycosis Lung, skin and mucous membrane, bone, joints
Aspergillus spp., particularly A. fumigatus Aspergillosis Lung, bronchial airways and sinus cavities, ear canal, eye (cornea)
Sporotrix schenkii Sporotrichosis Granulomatous pneumonitis (rare), skin, joints, central nervous system, eyes
Mucorales, zygomycetes Mucormycosis Nose, sinuses, eye, lung (brain and other organs), gastrointestinal system
a
Sources: 112, 138.

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TABLE 5 Immunopathological responses caused by fungal hypersensitivity


Type Immune response Diseases
I: immediate hypersensitivity IgE, mast cell Asthma, rhinitis, eczema, and hay fever
III: immune complex IgG, antigen/antibody complexes deposition Hypersensitivity pneumonitis, Arthus
mediated in the blood vessels and tissues reaction, extrinsic allergic alveolitis
IV: delayed-type Antigen-sensitized T-lymphocyte Allergic contact dermatitis, pneumonitis
hypersensitivity

regions. However, when types of mold and levels that are (176). This method provided alternative tools to the
“atypical” in the indoor environment increase because of morphology-based methods for indoor mold research and
recurrent water leaks, home dampness, and high humidity, investigation. However, ERMI is still subject to debate and
the prevalence of allergy and respiratory problems also rises further evaluation and validation. It is questionable whether
(29, 39, 40, 52, 155–161). Dampness and mold have, in ERMI should be applied to nonresidential environments. It
many epidemiological studies, been shown to be associated has not been widely accepted as a valid exposure marker at
with cough, environmental lung disease, and asthma (116, present.
162–166). Molds found on wet building materials and known In clinical allergy studies, patients can be tested for specific
to be associated with allergy and respiratory problems are spe- mold allergy using skin or serological tests (IgE-RAST), and
cies of Alternaria, Aspergillus, Aureobasidium, Cladosporium, appropriate advice and treatment can then be prescribed. Due
Fusarium, Paecilomyces, Phoma, Penicillium, Rhizopus, Stachy- to the low sensitivity of some of the commercially available
botrys, Trichoderma, and others (7, 79, 97, 101, 167, 168). mold extract tests, false negative results are not uncommon.
The prevalence of allergy to fungi among atopic patients is Patients with an atopy are frequently allergic to multiple fun-
estimated to be around 30% and in the general population up gal species and manifest type I reactions (Table 5). Another
to 6%. Exposure to molds during childhood is suspected to be practical problem is that available mold extracts at present
a risk factor for development of allergic respiratory disease only cover a very small portion of molds we are exposed to
(169, 170), although a possible protective effect has been and that are common in buildings with dampness and water
reported concerning children’s microbial exposure in farm damage.
environments (171–173). New onset of asthma in children Most of the fungi in bioaerosols may be allergenic depend-
after prolonged moisture and mold exposure has been demon- ing on the exposure situation and doses (114), although the
strated in a prospective study (170). However, respiratory sensitivity of clinical tests may vary with the study population
problems can occur in atopic and nonatopic individuals and individual immune system characteristics. Atopic indi-
(atopy is a genetic trait of increased allergen sensitivity). viduals typically have a higher rate of positive skin reactions
The reported percentages of populations allergic to molds after provocation tests and serological allergy tests measuring
vary from 2% to 18%. Approximately 80% of asthmatics were antibody precipitins (IgE). Diseases such as allergic broncho-
reported to be allergic to molds (29). In a 2002 study, up to pulmonary aspergillosis (138) and allergic fungal sinusitis
35% of newly diagnosed asthma cases were attributable to possibly require additional host factors which are not well
workplace mold exposure (172). A recent study in Europe documented (144), and may be the result of a combined reac-
found that workplace exposure to dampness and molds is tion of allergenic inflammation and the immunotoxic effect
associated with the occurrence of new-onset asthma. In of fungal metabolites. The relevant route of exposure is inha-
addition, exposed workers suffering from asthma-like symp- lation. Fungal by-products (i.e., mycotoxins) have ciliostatic
toms are subject to an increased risk for the development of effects in the respiratory tract (177), which can be one of the
asthma (174). important pathological mechanisms causing diminished
The incidence and prevalence of allergic diseases is on the mucociliary clearing and local inflammatory effects in the air-
rise (114). Many patients with chronic rhinosinusitis have a ways and sinuses. In general, the adverse effects of fungal
very high incidence of positive fungal cultures (up to 96%), inhalation are related to duration and intensity of fungal
and it is often associated with allergic fungal sinusitis (144). exposure. However, typical for allergic reactions is that once
Park et al. (145) found building-related (BR) rhinosinusitis an individual develops an allergy to certain fungi, even small
symptoms were a risk factor for the onset and development airborne concentrations can trigger an asthma attack or other
of BR asthma symptoms and exposure to molds in water dam- allergic reactions. This is principally different from fungal
aged buildings is an increased risk for the development of BR toxic-inflammatory health reactions, which depend on air-
asthma symptoms among the individuals with BR rhino- borne concentrations and will be similar for most people,
sinusitis symptoms. Murr et al. (175) found that Alternaria whether or not they are sensitized. Allergy “threshold levels”
alternata, Cladosporium cladosporioides, Cladosporium herba- to common mold have been reported (151), but variations in
rum, Penicillium brevicompactum, Penicillium crustosum, and sampling strategies and methodological limitations make
Penicillium chrysogenum were at very high concentrations these very unreliable in practical settings (178, 179). There-
ranging from 1,451 to 2,867,839 cells equivalence/sample fore, the consensus is that acceptable safe threshold limits for
in the sinus samples of some chronic rhinosinusitis (CRS) fungal indoor exposure cannot be established (17, 180), and it
patients. However, the Environmental Relative Moldiness is generally recommended to avoid or minimize unnecessary
Index (ERMI) results did not show significant difference in indoor fungal exposures (181).
fungi in the dust samples between homes of CRS and Although a low rate of IgE-mediated allergic responses to
non-CRS patients. the toxigenic fungi Stachybotrys chartarum have been reported
ERMI is a DNA-based method, based on a collection in some studies, it is unlikely a strong allergen in the clini-
of approximately 1,000 dust samples collected from U.S. cal setting, and the toxic-irritant effects appear to be more
houses, developed to screen indoor environments for molds important (182). Chun et al. (183) established a suggested

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TABLE 6 Fungal agents of HP and occupational dust exposure


Fungal agent Sources Disease
Aspergillus clavatus Moldy malt Malt worker’s lung
Aureobasidium pullulans Steam Sauna-taker’s lung
Alternaria spp. Wood Wood worker’s lung
Botrytis cincerea Moldy fruits Winegrowers’ lung
Cryptostroma corticale Wood Maple bark stripper’s lung
Farnai rectivirgula Straw Potato riddler’s lung
Serpula (Merulius) lacrymans Moldy building Dry rot lung
Penicillium spp. Cork Suberosis, woodman’s disease
Penicillium casei Cheese Cheese worker’s lung
Mucor stolonifer Moldy paprika Paprika worker’s lung
Trichosporon cutaneum House dust Japan summer pneumonitis

threshold dose (10 μg) for S. chartarum allergy induction by (agriculture) by the National Institute for Occupational
comparing the allergenicity of S. chartarum to house dust Safety and Health. The measures include the use of industrial
mite extracts in a mouse model. They opined that exposure hygiene controls, special protective equipment, ventilation,
to S. chartarum might be easily over the sensitization thresh- and respiratory protection (192). Although ODTS is more
old for a susceptible population in buildings with damp or likely to occur in settings where large amounts of organic
water-damaged conditions. waste are handled (such as agriculture and composting
Nagayoshi et al. (184) reported for the first time that facilities), it may also happen in office and domestic environ-
inhalation exposure to the conidia of S. chartarum resulted ments (189).
in the remodeling of pulmonary arteries and pulmonary Large-scale composting of organic waste is a new, growing
hypertension in mice. Yike and Dearborn (185) considered technology in municipal waste management (191). Environ-
it a significant step to understand the pathologic effects of mental monitoring suggests that this procedure involves risks
S. chartarum. Rakkestad et al. (186) demonstrated that heat- of high levels of exposure to several pathogenic fungi and
treated S. chartarum conidia induced cell death (apoptosis) bacteria. Immunological blood changes (such as IgG anti-
within 3–6 h due to DNA damage. body elevation) can be observed in waste-handling workers
or other occupations with high fungal exposure (193).
Hypersensitivity Pneumonitis and Organic Cladosporium cladosporioides, Cladosporium sp., and Fusa-
Dust Toxic Syndrome rium napiforme were reported to cause HP in residences
The clinical features, biochemistry, and pathophysiology of (194–196). HP caused by fungi was recently reviewed by Sel-
allergic or inflammatory-toxic reactions to airborne microbial man et al. (197). Bhan et al. (198) indicated that Stachybotrys
exposure are difficult to separate (187, 188). Hypersensitivity chartarum is a potential cause of HP which is TLR9-depend-
pneumonitis (HP), also called extrinsic allergic alveolitis, is a ent in mouse model.
well-recognized occupational disease. Table 6 lists several
fungi, their sources, and the occupational hypersensitivity Mycotoxins and Human Health
diseases they cause. Organic dust toxic syndrome (ODTS), Fungi produce toxic chemicals, such as the poisonous
also called toxic pneumonitis, is a nonallergic, noninfectious compounds found in some mushrooms and the toxic metab-
form of an acute inflammatory lung reaction to high fungal olites of some species of microfungi. Many of the mushroom
dust exposure (89, 189, 190). The differences between HP poisons are polypeptides or amino acid–derived toxins (42).
and ODTS can be difficult to distinguish. Table 7 lists com- Poisoning due to ingestion of poisonous mushrooms is
parative features of HP and ODTS. The significance of excluded from the scope of this chapter, and the reader
ODTS in occupational health is such that preventive meas- should consult suggested references on mushroom toxins
ures have been recommended for certain occupations (199, 200).

TABLE 7 Comparative features of HP and ODTSa


HP (extrinsic allergic alveolitis) OTDS (toxic pneumonitis)
Immune responses Type IV delayed hypersensitivity, cell-mediated Nonallergenic, noninfectious, lack of IgG.
immune reaction
Affected tissue/organ Lung alveoli, forming granulomas Inflammatory lung reaction
Exposure levels 106–1010 CFU/m3 of thermophilic actinomycetes or High concentrations of fungi, >109 spores/m3,
fungi >1–2 µg/m3 of endotoxins, or (1–3)-ß-D-glucan.
Clinical features Dypnea, cough, fatigue, poor appetite, weight loss, Dypnea, cough, headaches, fever, chills, malaise,
abnormal chest X ray, abnormal pulmonary acute inflammatory lung reaction, negative chest X
functions, high antibody precipitins; may cause ray; may recover after exposure cessation
pulmonary fibrosis long term
a
Source: 191.

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Some fungi have been known to produce secondary knowledge of these adverse health effects has led internation-
metabolites called mycotoxins that are harmful to animals ally to regulatory efforts to protect humans from excess expo-
and humans (19–21), and specifically when ingested (201, sure in food and agricultural products based in many cases on
202), inhaled (5, 156, 203, 204), or in contact with the a “precautionary principle,” in part because the data are still
skin (205–207). Mycotoxins in food and feed may lead to limited and definite dose-response models have not yet
cancer and mutagenicity, and estrogenic, gastrointestinal, been established for these agents. Toxic molds can induce
and kidney disorders. Some mycotoxins are immunosuppres- abortions and reproductive abnormalities in animals. There
sive and compromise the resistance to infectious disease. In is a concern among environmental health clinicians about
food safety mycotoxins are well recognized and regulated as similar effects in humans, however adequate human studies
potential disease agents affecting human and animal health are lacking. The International Agency for Research on Can-
(see “scientific opinion” at http://www.efsa.europa.eu/en/ cer (IARC) classified aflatoxin produced by Aspergillus flavus,
topics/topic/mycotoxins.htm). A. niger, and A. parasiticus as having “sufficient evidence” for
A recent study found that settled dust collected from human and animal liver carcinogenicity (217). Ochratoxin A
moisture-damaged, damp schools contained larger numbers is a potent nephrotoxin with immunosuppressive, terato-
of microbial secondary metabolites at higher levels than the genic, and carcinogenic properties and has been classified
dust samples from schools without moisture damage and as a possible carcinogen to humans (85, 221). Sterigmatocys-
dampness (208). Mycotoxin production is species specific tin is also a carcinogen (224). Whether patulin is carcino-
(22, 209). Nielsen and Frisvad (22) pointed out the six chal- genic is subject to further research. A recent study showed
lenges faced in mycotoxin research at present. Among the the possible role of free radicals in patulin-mediated dermal
challenges, flaws in methodology and misidentification of tumorigenicity involving mitogen-activated protein kinase
mycotoxigenic fungi or using sequences generated from mis- (222). Fumonisins (B1 and B2) are cancer-promoting metab-
identified cultures had led to some questionable results, olites (223, 224). Many of the other mycotoxins have not
such as false negative or positive results. The most important been classified as carcinogens, but carcinogenicity cannot
one among these hurdles is expertise. be ruled out due to a lack of appropriate studies.
These mycotoxins belong chemically to the alkaloids, Human cases of true mycotoxicosis appear to be rare and
cyclopeptides, and coumarins (3). Early reports of the delete- in the past were thought to be mostly related to ingestion of
rious and poisonous effects of mycotoxins on human health contaminated grain products. However, possible occupa-
goes back to the 1100s on ergotism (holy fire or St. Anthony’s tional or environmental inhalation exposures have been
fire) caused by consumption of rye bread contaminated by the described in recent case studies and epidemiological surveys.
alkaloid-containing ergot developed by Claviceps purpurea Typical nonallergic symptoms of patients in which myco-
(210). At present, more than 400 mycotoxins have been dis- toxin exposure was either confirmed or highly suspected are
covered (207). The total number of mycotoxins remains recurrent cold and flu–like symptoms, extreme fatigue, con-
unknown but is believed to be in the thousands. stant sore throat or skin irritation, severe and unusual head-
The effects of toxins produced by molds in humans have aches, neuromuscular and neurocognitive dysfunction
been mostly described and researched in relationship to food- (tremor and shakes, unusual memory and concentration prob-
borne diseases affecting animals or regional human disease lems), bleeding disorders of the lung in infants, irregular
outbreaks (202, 211). The earliest known mycotoxin pro- menses, diarrhea, dermatitis and irritation of skin, and
ducers, primarily Claviceps purpurea, produce the substance impaired immune function (23, 225–229). Mycotoxins may
ergot, which causes ergotism. Ergot toxins caused food poi- also be involved in occupational diseases and respiratory can-
soning outbreaks due to the consumption of contaminated cers among food and grain workers (230–232). Environmen-
rye bread. The toxin was associated with bizarre behaviors tal sentinel investigations in water-damaged buildings have
(known as “dancing plague,” “holy fire,” or “St. Vitus’s shown detectable levels of airborne mycotoxins from Stachy-
dance”) and may have contributed to the population decline botrys chartarum and others that may be of concern (233–
in Europe from the fourteenth to the eighteenth centuries 236). This is important in clean-up and remediation projects,
(210, 212). as the removal of toxins from water-damaged and moldy
There has been a debate regarding the public health building materials is difficult (237). Further studies are
importance of “toxic mold” in enclosed indoor environments needed to improve our understanding of mycotoxins found
and its impact on the occupants’ health (213, 214). Aspergil- in the indoor environment and possible adverse human
lus versicolor and species of Penicillium, Fusarium, Trichoderma, health effects.
Cephalosporium, Chaetomium, and Stachybotrys are known to Some mycotoxins, such as lysergic acid, are derivatives of
produce naturally potent mycotoxins, depending on available amino acids (such as tryptophan). Others derived from other
nutrients, favorable environmental conditions, or their life precursors are grouped into aromatic and phenolic–related
cycle. Health complaints and clinical findings in patients liv- toxins and terpenoid toxins. Some well-known and potent
ing or working in wet and moldy buildings often cannot be mycotoxins in the aromatic and phenolic–related toxin group
explained as allergic reactions in otherwise healthy individu- are aflatoxin, zearalenone, and griseofulvin. The terpenoid
als. An overview of clinically important health disorders toxins include trichothecenes and fusidanes (3). There
based on various case reports and results of disease cluster were more than 200 mycotoxins produced by a variety of com-
investigations are presented for the most important myco- mon fungi according to the WHO Environmental Health
toxin producers (Table 8) (215–217). Criteria 105 on mycotoxins, published in 1990 (238). Sam-
The toxicological knowledge of such mycotoxins pri- son (127) suggested that there were more than 400 toxic
marily stems from the veterinary and food-safety science metabolites in 1992. It is likely that the number of recorded
(218). A limited number of the more than 400 known myco- toxic metabolites will increase over time because of new dis-
toxins have been studied and found to have important geno- coveries (216, 239, 240). These alcohol- and water-soluble
toxic, mutagenic, cytotoxic, carcinogenic, nephrotoxic, toxins can be attached to spores, mycelia, or dust particles
pseudo-estrogenic, immunosuppressive, protein synthesis and are sufficiently small in size (2–10 micron) to be inhaled
inhibitor, or other toxic properties (19–21, 219, 220). The into the human lungs. Some mycotoxins are lipid soluble and

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TABLE 8 Some toxigenic fungi, fungal chemical metabolites, and their health effects
Fungi Chemical metabolites Health effects
Penicillium (>200 species) Patulin Hemorrhage of lung, brain disease
citrinin Renal damage, vasodilatation, bronchial constriction,
increased muscular tone
ochratoxin A Nephrotoxic, hepatotoxic
citroviridin Neurotoxic
emodin Reduced cellular oxygen uptake
gliotoxin Lung disease
verruculogen Neurotoxic: trembling in animal
secalonic acid D Lung, teratogenic in rodents
Aspergillus spp. Patulin Hemorrhage of lung, brain disease
A. clavatus Aflatoxin B1 Liver cancer, resp. cancer, cytochrome P-450
monooxygenase disorder.
A. flavus and A. parasiticus Sterigmatocystin Carcinogen
A. versicolor ochraceus Ochratoxin A Nephrotoxic, hepatotoxic
Stachybotrys chartarum, Trichothecenesa (more than 50 derivatives Immune suppression and dysfunction, cytotoxic,
Fusarium species, known): T-2, nivalenol, deoxynivalenol, bleeding, dermal necrosis; high dose ingestion = lethal
Trichoderma species diacetoxyscirpenol, satratoxin H, G, (human case reports); low dose, chronic = potentially
other macrocyclic trichothecenes lethal); teratogenic, abortogenic (in animals).
Spirolactone Hemorrhage
Zearalenone “Alimentary toxic aleukia” (ATA) reported in Russia
and Siberia
“Staggering wheat” in Siberia
“Red mold disease” in Japan
Neurotoxic/nervous behavior abnormality
Co-carcinogen/chemotoxic (?)
Anticomplement function.
Phytoestrogen may alter immune function; stimulates
growth of uterus and vulva, atrophy of ovary
Claviceps spp. Ergot alkaloids Prolactin inhibitor, vascular constriction, uterus
contraction promoted.
a
Trichothecenes are also produced by Myrothecium, Trichothecium, and Gibberella (teleomorph of some Fusarium species).

may be absorbed via skin. Building materials contaminated is often neither allergic nor infectious in nature. In several dis-
with the above-mentioned fungi have been shown to produce ease outbreaks, human and animal death has been linked to
detectable levels of mycotoxins on the materials (97, 167, exposure to toxigenic fungi, typically through ingestion. Cur-
168, 241). rent research, however, indicates that inhalation of certain
Aflatoxin may be involved in occupational respiratory mycotoxins has even stronger effects (247) and may be fre-
cancers among food and grain workers (242). Aflatoxin- quently associated with health complaints and human dis-
induced disease has been well reviewed (217, 243, 244). Tri- ease. The occurrence of mycotoxins in the environment,
chothecene toxins (T-2 toxin, Fusarium toxins) are listed their chemistry and related adverse health effects have been
with “limited evidence” for animals and “inadequate evi- reviewed (5, 20, 21, 127, 206, 216, 248). Several mycotoxins,
dence” (no data available) for humans (217). Macrocyclic even in low concentrations, were observed to be cytotoxic,
trichothecenes, such as satratoxin H, have not been classified. interfere with DNA and RNA synthesis, inhibit protein syn-
Epidemiological studies suggest a higher rate of upper respira- thesis, and cause apoptosis of cells of different body organs.
tory tract and lung cancer in workers in the grain and food- These toxic effects may cause a variety of short-term as well
handling industry with high fungal product inhalation risk as long-term adverse health effects in animals and humans
(230). The high rate of lung cancer among uranium miners (21, 168, 248–250). Samson (127) divided the effects into
in Slesien (Schneeberg disease), may be related to combined four basic categories: acute, chronic, mutagenic, and terato-
effects of high radon and Aspergillus exposure in the under- genic. Symptoms thought to be due to mycotoxins or toxin-
ground mines (244). containing spores ( particularly those of S. chartarum) include
Research has shown that water-damaged building materi- dermatitis, recurring cold and flu–like symptoms, burning
als are often contaminated with fungi that produce detectable sore throat, headaches and excessive fatigue, diarrhea, and
levels of mycotoxins (157, 167, 245, 246), which may impaired or altered immune function (206). The ability of
become airborne and further contribute to indoor air pollu- the body to resist infectious diseases may be weakened, result-
tion (101, 241). From a public health point of view, probable ing in opportunistic infections. Certain mycotoxins, such as
important toxigenic fungi are Aspergillus species, Penicillium zearalenone, have been found to cause infertility and still-
species, Fusarium species, S. chartarum (syn. S. atra), Paecilo- births in pigs (201). Low-level, complex exposures from a
myces species, and Trichoderma species. These fungi have mixture of mycotoxins, as would be typically encountered
been associated with adverse health effects in humans and in real-life situations, may have synergistic effects, which
animals resulting in typical organ damage and disease, which may result in central neuroendocrine-immune changes and

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consequently in complex effects of the endocrine and nerv- mycotoxins were in airborne fungal propagules of S. atra
ous system (121). and could be collected on membrane filters. Conidia of
Nonallergic complaints from patients in which myco- A. flavus and A. parasiticus were reported to contain aflatoxins
toxin-producing fungal exposure was either confirmed or (263). Miller (5) also reported detection of two mycotoxins,
strongly suggested include recurrent cold and flu–like symp- deoxynivalenol and T-2 toxin, in conidia of Fusarium grami-
toms, extreme fatigue, constant sore throat or skin irritation, nearum and F. sporotrichioides, respectively. These references
severe and unusual headaches, neuromuscular and neurocog- suggest that inhalation exposure to conidia may also increase
nitive dysfunction (tremor and shakes, unusual memory and the chance of exposure to mycotoxins. Corey et al. (123)
concentration problems), bleeding disorders of the lung in found that satratoxin G from S. chartarum induced rhinitis
infants, irregular menses, diarrhea, dermatitis and irritation and apoptosis of olfactory sensory neurons in the nasal airways
of skin, and impaired immune function. Mycotoxins may of rhesus monkeys.
also be involved in occupational respiratory cancers among Although relationships were established to link inhalation
food and grain workers (230), however, typical home or office exposure to mycotoxin-containing fungal spores and symp-
indoor environments have not been studied. Better con- toms of mycotoxicoses in fungi-infested indoor environments
trolled studies are needed to improve our understanding of (155, 203, 261, 262), other possible exposure routes such as
mycotoxins found in the indoor environment and possible ingestion and skin contact are likely. Because fungal spores
adverse human health effects. are ubiquitous in a contaminated environment, the chance
Historically, mycotoxins have been a problem to farmers of ingesting toxin-containing spores is likely to increase
and food industries and in Eastern European and developing through eating, drinking, and smoking. It is prudent to limit
countries (251). The large-dose exposure to fungi and myco- exposure to such potent toxic chemicals (124), particularly
toxins encountered by farmers and in food industries was gen- when significant fungal growth and amplification is found
erally considered unlikely to occur in nonfarming activities. indoors (134).
However, many toxigenic fungi, such as S. chartarum and spe- The toxigenic fungus frequently detected in “problem
cies of Aspergillus, Penicillium, and Fusarium have been found buildings” is S. chartarum, which produces a series of potent
to infest buildings with known indoor air and building-related cytotoxins (trichothecenes, satratoxins, and spirolactones)
problems and illnesses (29, 156, 203, 252). It has been sug- as well as a variety of other compounds affecting the immune
gested that inhalation exposure to mycotoxin-containing system (240, 264, 265). Case studies of health effects and
fungal spores is significant in the reported cases of building- immunological laboratory changes related to indoor exposure
related mycotoxicoses (252). Croft et al. (203) reported sev- to trichothecenes and possibly other mycotoxins, disorders of
eral cases of mycotoxicoses caused by airborne exposure to the the respiratory and central nervous system were noted (215,
toxigenic fungus S. chartarum in a residential building. Addi- 266, 267). Abnormal test results of the cellular and humoral
tional cases of office building–associated Stachybotrys myco- immune system were found (152). In earlier cases in Eastern
toxicosis were reported by Hodgson et al. (226), Johanning Europe, typically in an agricultural setting, marked leukope-
(253), and Johanning et al. (156). Satratoxin H was detected nia or acute “radiation-mimetic” effects on the blood cell sys-
in the fungus isolated from the contaminated building. Hem- tem with subsequent sepsis-like opportunistic infections after
orrhagic lung disease in infants was highly associated with trichothecene ingestion were reported (268, 269).
indoor S. chartarum exposure in a case cluster investigation Trichothecenes are considered to be the most potent
in Cleveland (254) and in a case-home investigation in small molecule inhibitor of protein synthesis, acting through
the U.S. Midwest (255). Subsequently consultants for the inhibition of the peptidyl transferase activity (219, 270).
Centers for Disease Control and Prevention called for more These toxins can cause alveolar macrophage defects and
research to prove the causal relationship of Stachybotrys may affect phagocytosis. They have been investigated for
and idiopathic pulmonary hemorrhage in infants on review use in cancer treatment (271), but also in chemical-biological
of the Cleveland study (256). After more than 10 years, warfare. The presence of fungal chemical metabolites has
more cases were reported, increasing from 9 cases in the orig- been reported in several cases of animal and human ingestion-
inal study to 52 cases at present. Among the cases, 91% of related mycotoxicosis, resulting sometimes in death (252,
patients were living in residences with S. chartarum (257). 272). Mycotoxins, such as satratoxin H of the trichothecene
Studies with toxic Stachybotrys fungi in mice showed similar group, have been shown to cause depressed T or B lymphocyte
effects (inflammation and hemorrhage) (258). S. chartarum activity, suppressed immunoglobulin and antibody produc-
was isolated from brochoalveolar lavage fluid of a child tion, reduced complement or interferon activity, and
with pulmonary hemorrhage (259), and S. atra exposure impaired macrophage-effector cell function of human neutro-
was found in an infant who developed laryngeal spasm and phils (20).
hemorrhage during general anesthesia (260). In an epide- Laboratory changes of immunoglobulins (IgA, IgE, IgG,
miological study a high prevalence of pulmonary diseases and IgM) in workers handling mycotoxin-contaminated
of office workers in Florida court buildings were reported foodstuffs, primarily deoxynivalenol (vomitoxin), have
after prolonged indoor exposure to primarily S. chartarum been reported (273). An increase of IgA production and
and A. versicolor (226). IgA nephropathy and a decrease of IgG and IgM after inges-
Mycotoxins generally have low volatility; therefore, inha- tion of vomitoxin were reported in a mice experiment (274).
lation of volatile mycotoxins is not very likely (206). Rather, Renal failure and IgG deposition in the glomeruli after inha-
the toxins are an integral part of the fungus. Sorenson et al. lation of ochratoxin produced by Aspergillus ochraceus was
(261) demonstrated in the laboratory that aerosolized conidia found in the case of a farmer (275).
of S. atra contained trichothecene mycotoxins. The most A WHO task group concluded that an association
common toxin was satratoxin H. Lesser amounts of satratoxin between trichothecene exposure and human disease episodes
G and trichoverrols A and B were also detected but less fre- is possible; however, only limited data are available (238).
quently. They also found that most of the airborne particles Immunotoxicological effects principally depend on the expo-
were within respirable range. Similar experiments, conducted sure conditions, dose, and timing. Some immunological
by Pasanen et al. (262), demonstrated that trichothecene effects may only be transient, of short duration, and difficult

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3.2.5. Airborne Fungi and Mycotoxins ▪ 3.2.5-13

to detect in routine medical tests. Medical findings are often damp and water-damaged buildings play an important role
nonspecific and other systemic diseases or causes need to be in public health and disease prevention. There is a growing
ruled out by the experienced clinician. The treating physician consensus among experts that fungi associated with dampness
often does not recognize mycotoxicosis, especially because leads to preventable health problems, primarily of the respira-
exposure circumstances and presence of certain mycotoxins tory organs and allergies. They are allergenic and irritant/
are unknown. Advanced fungal exposure characterization toxic agents that typically cause or aggravate airways, or are
and sampling techniques now available should improve the associated with infectious and mainly respiratory diseases in
chances for better medical detection of mycotoxicosis. exposed people. Research findings indicate that they are a
Analytical methods involving immunoassays and cell line major problem in buildings where moisture control is poor
cytotoxicity analysis are able to provide relatively rapid and or where water intrusion is common. Human exposures in
easy screening tests to detect the presence of mycotoxins in these situations typically are a mixture of different fungi and
fungal-contaminated materials (157, 250, 276). bacteria. Synergistic inhalation effects of fungal by-products,
Mycotoxin research faces methodological challenges, such as mycotoxins in fungal spores, β-glucans, or likely fun-
such as misidentification of mycotoxigenic fungi or using gal MVOCs released into the surroundings are potentially
sequences generated from misidentified cultures (22). This irritating, toxic, teratogenic, carcinogenic, and immune-
may led to erroneous results, such as false negative or positive suppressive. Clinical diagnoses of mold allergies and fungal
results. infections are generally easier and less complicated than
emerging health concerns of such fungal metabolites as
Volatile Organic Compounds Produced by Fungi (1-3)-ß-D-glucan, airborne mycotoxins, and fungal MVOCs.
Fungi in active growth produce VOCs also known as Furthermore, risk assessment of human exposure to these
MVOCs, which typically are noticed as a musty, moldy fungi and their by-products is complex, because multiple
odor. Over 200 MVOC compounds have been identified agents, hypersensitivity reactions, and different disease out-
from different fungi (277). Indoor measured VOC levels, comes are involved. Human susceptibility to them varies
however, are typically low and any serious health risks are from individual to individual. Some of the health implica-
uncertain (278, 279). Possibly, related mucous membrane tions from inhalation exposure of fungi are undergoing further
and olfactory irritations may trigger an “unpleasant odor reac- research, particularly at low exposure concentrations that are
tion” and annoyance. Measurement of VOCs may be an indi- likely different from agricultural settings studied in the past.
cator of excessive indoor fungal growth (280). A number of In most cases, diligent exposure cessation and control leads
VOCs have been identified from fungi common in indoor to symptom reversal and health improvement. Little is known
contamination. Most of these fungal VOCs are derivatives concerning the consequences of short-term and long-term
of alcohols, aldehydes, amines, ketones, terpenes, esters, environmental exposures to mycotoxins and whether all of
hydrocarbons, aromatics, and sulfur-containing compounds the health effects are reversible. However, based on what is
(281, 282). The in vitro production of fungal volatiles from known at present regarding fungal exposures and potential
47 Penicillium taxa were made up of alcohols, ketones, esters, adverse health effects, it is prudent to avoid or minimize expo-
small alkenes, monterpenes, sesquiterpenes, and aromates sure to infectious, allergenic, and toxic fungi and to control
(283). However, aldehydes were not detected. indoor growth conditions. Furthermore, fungal growth
Some of the fungal VOCs have an unpleasant odor (4), indoors suggests water infiltration and damage to building
and other fungi (such as mushrooms) produce VOCs of pleas- structures and material, that ought to be corrected to avoid
ant odors and flavors. 1-Octen-3-ol, one of the major fungal further decay.
VOCs, has a characteristic mushroom odor. The musty,
moldy, and earthy odors are likely to come from 2-octen-1-ol
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