APPLIED AND ENVIRONMENTAL MICROBIOLOGY, Nov. 1994, p. 4172-4173 Vol. 60, No.

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0099-2240/94/$04.00+0
Copyright C 1994, American Society for Microbiology

Fungal Production of Volatiles during Growth on Fiberglass

I. M. EZEONU,' D. L. PRICE,2 R. B. SIMMONS,' S. A. CROW,' AND D. G. AHEARN"*
Biology Department, Georgia State University, Atlanta, 1 and Interface
Research Corporation, Kennesaw,2 Georgia
Received 16 May 1994/Accepted 7 September 1994

Acoustic and thermal fiberglass insulation materials used in heating, ventilation, and air-conditioning
systems were colonized with fungi in laboratory chambers. The mixed fungal population, principallyAspergillus
versicolor, Acremonium obelavatum, and Cladosporium herbarum, produced odoriferous volatiles, including
2-ethyl hexanol, cyclohexane, and benzene. These volatiles may be related to poor indoor air quality and the
sick building syndrome.

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More than 300 volatile organic compounds (VOCs) have 15 g) from laboratory chambers or directly from buildings were
been identified in indoor air (8, 9, 12). The chief sources of packed into separate trap vessels and held for 7 to 14 days at
VOCs in indoor air are thought to be building materials, 22 to 25°C. The headspace air in the trap vessel was drawn
combustion processes, consumer products, and other synthetic through a desorption tube packed with Carbotrap C (3 cm),
materials used in homes and office buildings (8, 9). A great Carbotrap (3.5 cm), and Carbosieve (1.5 cm) sorbent charcoals
variety of volatiles, most of which are aliphatic alcohols, (Suppelco Inc., Bellefonte, Pa.). The charcoal bed containing
ketones, and terpenes, are produced by fungi (2, 3, 5). Some of the adsorbed organic compounds was purged with helium and
the more commonly identified VOCs of fungi include geosmin, thermally volatilized and cryofocused with a Chrompack TCT
3-octanone, 2-octen-1-ol, 3-methyl-1-butanol, and 3-methyl- unit (Raritan, N.J.). The captured compounds were thermally
furan (2, 3, 5, 6, 10, 15). The various volatiles reported have desorbed at 250°C and injected onto a Hewlett-Packard GC
been mainly from fungi grown in culture, or culture findings column (50 m by 0.2-mm internal diameter; film thickness, 0.33
have been correlated with analyses of building air (1-3, 5). The mm). The detection was accomplished with a Hewlett-Packard
number and types of VOCs produced have been greatly 5971A mass selective detector in a scan mode with a range of
influenced by the substrate and conditions of growth of the 30 to 300 atomic mass units. Spectra were analyzed and peaks
fungi (1, 3, 11). In laboratory chambers, Acremonium obclava- were identified with the NIST/EPA/MSDC 49K Mass Spectral
tum was the most common colonizer of new unused fiberglass Database (U.S. Department of Commerce, Washington,
duct liners and duct boards (13), whereas Aspergillus versicolor D.C.). The detection limit for organics was 50 mg/m3, based on
was most common in samples from various "sick buildings" a toluene (1-methyl benzene) external standard. Identifications
(buildings in which occupants complained of moldy air) (7).
The urea-formaldehyde resins that bind the fibers as well as the
acrylic-latex facings of the duct liners were indicated as prob-
able nutrient sources for the growth of fungi (7, 13). A. TABLE 1. VOCs produced by fungi in pure culture on enriched
versicolor is a known producer of the mycotoxin sterigmatocys- agar and from colonized fiberglass insulation materialsa
tin (14) and of a variety of volatile metabolites including VOCs produced on:
geosmin, 3-octanone, 3-methyl-furan, 2-methyl-1-butanol, Species Colonized fiberglass
2-methyl-1-propanol, dimethyl benzene, ethyl benzene, and Enriched agar duct liner"
2-ethyl hexanol (2, 4, 5). This last compound is a known eye
irritant with an unpleasant odor and may be associated with A
some of the skin and upper airway irritations found in the sick A. obclavatum Ethanolc (91)d, acetoneC Cyclotrisiloxane (91),
building syndrome (3). Herein, we examine the production of (78), 2-butanone (86), limomenec (91),
volatile metabolites by some colonizers of fiberglass insulation methyl benzenec (87), pentane (90),
materials on enriched media as well as on the natural substrate cyclohexanec (96), 2-ethyl arsenous acid (91),
(fiberglass duct liner). hexanol (72), benzenec benzenec (90)
Ten- to fifteen-gram sections (6 by 12 cm) of fiberglass duct (60)
B
liner were exposed to fungal colonization in laboratory cham- A. versicolor 1,3-Dimethoxy benzene Methyl benzenec (91),
bers for a period of 28 to 90 days at relative humidities of about (94), methyl benzenec cyclotrisiloxane
95% (7, 13). The fungi were identified from acetate tape (90), cyclotetrasiloxane (91), ethanolc (86),
mounts of the colonized surfaces, and sections of colonized (94) xylene (64), D-
materials were cultured on mycological agar (Difco) with limonenec (91), 2-
0.05% chloramphenicol for verification of identification. ethyl hexanol (78),
Cultures were grown on mycological agar in 100-ml boro- benzeneC (83)
silicate purge-and-trap vessels (Wheaton, Millville, N.J.) for 5
to 7 days at 22 to 25°C. Colonized fiberglass insulations (10 to a Only volatiles with a high probability of accurate library identification are
listed.
*Colonized by a mixed population of A. obclavatum and A. versicolor,
Cladosporium herbarum was also present. These volatiles were not detected from
noncolonized duct liner.
*
Corresponding author. Mailing address: Biology Department, c Confirmations were made by comparisons with external standards.
Georgia State University, P.O. Box 4010, Atlanta, GA 30303-4010. d The numbers in
parentheses are quality of assurance values for gas chroma-
Phone: (404) 651-3110. Fax: (404) 651-2509. tography-mass spectrometry.

4172
VOL. 60, 1994 NOTES 4173

. (A) onstrate that fungi in heating, ventilation, and air-conditioning

systems may play a significant role in health problems encoun-
tered in some sick buildings.

i
0

isI i This research was supported in part by a contract awarded to

Georgia State University by NASA and by National Institute of
General Medical Sciences award 1 F31 GM16377-01 SRC-1.

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