Aerosol Science and Technology

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Aerodynamic Diameters and Respiratory

Deposition Estimates of Viable Fungal Particles in
Mold Problem Dwellings

T. Reponen

To cite this article: T. Reponen (1995) Aerodynamic Diameters and Respiratory Deposition
Estimates of Viable Fungal Particles in Mold Problem Dwellings, Aerosol Science and Technology,
22:1, 11-23, DOI: 10.1080/02786829408959724

To link to this article: https://doi.org/10.1080/02786829408959724

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Aerodynamic Diameters and Respiratory
Deposition Estimates of Viable Fungal
Particles in Mold Problem Dwellings
T . Reponen
Kuopio Regional Institute of Occupational Health,
P. 0 . Box 93, FIN-70701 Kuopio,
Finland
The aerodynamic size distributions of particles of ma-
jor fungal genera or groups, i.e., Penicillium, Cladospo-
rium, Aspergillus, and yeasts, were studied in real
exposure situations in mold problem and reference
dwellings. A new calculation method was used to esti-
mate respiratory deposition of fungal particles on the
basis of the measured data. The major fungal genera
had their maximum concentrations in the size range
2.1-3.3 pm, where alveolar deposition for particles
> 0.5 p m also has a maximum. According to respira-
tory deposition calculations for the most obvious
breathing patterns in the home environment, 30%-50%
INTRODUCTION
Fungal particles, consisting of fungal
spores, hyphal fragments or cells, such as
yeast cells, are organic particles ubiqui-
tously present in indoor environments. In
dwellings and office buildings, exposure to
airborne fungal particles may become a
problem due to an intramural source of
fungal particles, such as mold growth due
to water damage or other excess damp-
ness. People occupying moldy buildings
have been reported to suffer respiratory
symptoms and other health effects, such
as dermatitis and eye irritation (Platt et
al. 1989, Dales et al. 1991, Brunekreef
1992, Husman et al. 1993). The causal
relationships between exposure and symp-
toms are still poorly understood. There-
fore, the importance of various factors
determining exposure needs to be better
understood. Aerodynamic particle size
Aerosol Sc~enceand Technology 22:ll-23 (1995)
O 1995 Amencan Association for Aerosol Rescerch
Published by Elaevier Scicncc lnc.
of fungal particles would he deposited in the nose and
30%-40% in the alveoli during nasal breathing, whereas
70% would be deposited in the alveoli during oral
breathing. More Aspergillus species spores were esti-
mated to deposit in the alveoli for occupants of mold
problem versus reference dwellings whereas for yeasts
the situation was reversed. Comparison of airborne
fungal particle concentrations and numbers of de-
posited particles between mold problem and reference
dwellings indicated that more accurate estimation of
exposure to fungal particles is possible if one considers
both particle concentration and size.
(d,) is a critical factor when evaluating
respiratory exposure to fungal particles.
Particle size determines to a great extent
where fungal particles deposit in the res-
piratory system and hence the type of
respiratory reaction the particles may
cause. Allergic rhinitis and asthma can
result from exposure to relatively large
( > 5 pm) fungal particles (e.g., Altemaria
spp.) deposited in the nasal or extratho-
racic regions of the respiratory system. On
the other hand, allergic alveolitis requires
small fungal particles ( < 5 pm, e.g., As-
pergillus spp.) that penetrate to the alveoli
(Lacey and Crook, 1988; Burge, 1989).
The physical diameters of fungal parti-
cles are well known, e.g., Penicillium
chtysogenum is 3-4 p m in length and
2.8-3.8 p m in width (Samson and van
Reenen-Hoekstra, 1988). However, d, de-
termines fungal particle transport in air,

as well as in human airways and sampling
devices (Baron and Willeke, 1993). Aero-
dynamic particle size can be calculated
when the density, shape and physical
diameter of a particle are known. The
densities of fungal particles common
outdoors vary from 0.56 to 1.44 g/cm3
(Gregory, 1973). However, there are no
reports on the densities of fungal particles
most common in indoor air. It appears
that neither fungal particle density, nor
physical diameter or shape are constant,
but vary according to the particle's state
of hydration. At high relative humidity,
fungal particles absorb water and their
size increases (Pasanen et al., 1991a;
Madelin and Johnson, 1992). At low hu-
midity, fungal particles lose water and may
decrease in size, partially collapse, change
shape or develop internal gas bubbles (In-
gold, 1956; Lacey, 1991; Madelin and
Johnson, 1992). The effects of hydration
and dehydration on fungal particles are
not well understood at this time and ap-
pear to vary widely among species (Lacey,
1991; Pasanen et al., 1991a).
The dyes commonly used to distinguish
fungal particles from other particles, in
microscopic analysis, cause fungal parti-
cles to inflate to their physical maxima
(Madelin and Johnson, 1992). When de-
termining fungal particle size by electron
microscopy dimensional changes also de-
pend on sample preparatory treatment
(Beckett et al., 1984). Therefore, for the
purpose of estimating fungal particle be-
havior in air and the respiratory tract,
methods that given d , directly are prefer-
able to size analysis based on microscopic
measurements. Microscopic analysis, how-
ever, is helpful in determining the degree
of aggregation of fungal particles.
Lacey and Dutkiewitz (1976) deter-
mined d , for fungal particles released
from hay in a sedimentation chamber. The
d , calculated from settling velocities, were
3.1 p m for Aspergillus furnigatus and
3.2 p m for mixed Penicilliurn spp. The
T. Reponen
physical diameter of A. fumigatus spores
is 2.5-3 p m (Samson and van Reenen-
Hoekstra, 1988). Pasanen et al. (1991a)
studied the behavior of viable fungal par-
ticles at different humidities (12%-73%)
with a six-stage impactor. These authors
studied fungal genera common both in-
doors and outdoors and reported the fol-
lowing aerodynamic diameters for the
lowest and highest humidity values, re-
spectively: 2.0 and 2.7 p m for A. furniga-
tus, 2.2 and 3.9 p m for Penicilliurn spp.,
and 1.6 and 4.6 p m for Cladosporiurn spp.
Madelin and Johnson (1992) used an
aerodynamic particle sizer (APS) and a
cascade impactor in the humidity range of
40%-98%, obtaining the following d ,
with the APS: 1.9-2.2 p m for A. furniga-
tus, 2.6-3.0 p m for Penicilliurn chlyso-
genurn, and 2.3-2.5 p m for Cladosporiurn
cladosporioides. The corresponding physi-
cal diameters are 2.5-3 pm, (3-4) X
(2.8-3.8) pm, and (3-7) X (2-4) pm, re-
spectively (Samson and van Reenen-
Hoekstra, 1988). Madelin and Johnson
also observed the largest aerodynamic
particle diameters at the highest humidity.
Comparison of APS and impactor mea-
surements revealed that aggregated fun-
gal particles in chains, behaved aerody-
namically similar to, or only slightly larger
than single fungal particles. Lacey (1991)
reported similar results.
All of these studies have been per-
formed in experimental settings with labo-
ratory-grown fungal particles. However,
the nutrients provided in growth media
(Harding, 1975; Ellis, 1981) as well as air
humidity affect fungal particle size and
shape. Therefore, it may be possible that
fungal particles that are released directly
from laboratory cultures differ in size from
fungal particles from natural sources and
have a different residence time in the air.
It is possible to estimate exposure to
airborne particles via the respiratory sys-
tem using respiratory deposition data if
particle concentration and aerodynamic
Fungal Particle Size and Respiratory Deposition
size distribution are known. There are
many reports available about regional
particle deposition in the respiratory tract,
based either on models (e.g., Morrow,
1966; Stahlhofen et al., 1989; Koblinger
and Hofman, 1990; James et al., 1991), or
on experimental data from humans (e.g.,
Heyder et al., 1986; Chan and Lippmann
1980) or animals (e.g., Raabe et al., 1977).
However, there is little exposure informa-
tion available based on combining d, and
particle concentration with respiratory de-
position data. Hewett (1991) suggested this
approach for occupational epidemiology,
and Jarabek et al. (1989) combined lung
deposition data with theoretical particle
size distributions to extrapolate from ani-
mal toxicity data to human health risk
assessment.
In this study, d, for fungal particles
have been studied in situ in real exposure
situations in moldy and nonmoldy refer-
ence dwellings. Fungal particle concentra-
tions, as well as a general analysis of the
aerodynamic size distributions of fungal
particles in these dwellings, have been
presented previously (Reponen et al.,
1994). In this paper, the size distributions
of individual genera were analyzed. A new
calculation method to estimate respira-
tory particle deposition has been intro-
duced and the fungal particle deposition
has been estimated from the measured
data.
MATERIAL AND METHODS
Measurement and Data Analysis of the
Size Distribution of Fungal Particles
The present study was carried out in 12
dwellings. Six of the dwellings had mold
problems and each was matched with a
reference dwelling with no visible signs of
mold problems. All dwellings had ventila-
tion systems of mechanical exhaust.
Samples of airborne fungal particles
were collected twice in each dwelling, in
13
fall 1990 and in winter 1991. Outdoor
samples were also collected in fall be-
cause outdoor fungal particle concentra-
tions may influence indoor concentrations
and flora. Outdoor air samples were not
collected because outdoor concentrations
of fungal particles are extremely low due
to snow cover in winter (Reponen et al.,
1992).
Samples were taken with a six-stage
impactor (Model 10-800, Graseby Ander-
sen, Atlanta, GA) on malt extract agar.
The method allows the measurement of
viable fungal particles, i.e., those particles
that are able to grow in the agar medium
used. The 50% cutoff diameters of the
sampler are: 0.65, 1.1, 2.1, 3.3, 4.7 and
7.0 p m (Andersen-Sampler, 1976). Yeasts
were counted separately and molds were
identified to genus using an optical micro-
scope to examine mycelial and fungal par-
ticle morphology. For detailed description
of the dwellings and sampling procedure
see Reponen et al. (1994).
The four most common sampled fungal
genera or groups were included in the
analysis of the size distributions. Size dis-
tributions of fungal particles were calcu-
lated as geometric means for each fungal
genera or group by using the transforma-
tion log(x + 1). First, the geometric mean
diameter, d,, was calculated for each fun-
gal genus in each sample. Then the geo-
metric mean of the d, values over all
samples, d ,,, was calculated for each
genus. Differences in size distribution of
fungal particles between mold problem
and reference dwellings as well as be-
tween fall and winter periods were studied
using the Wilcoxon test. The d, values for
different fungal genera were compared
with the Friedman test (Siegel, 1988).
The aerodynamic diameters of selected
indoor air fungi were calculated for the
lowest and highest possible physical sizes.
The effects of fungal particle aggregation
and density on d, were calculated for P.
chpsogenum as an example, following

Hinds's (1982) method and using Fuchs's
(1964) dynamic shape factors for single
particles and Davies's (1979) factors for
aggregated particles. Equivalent volume
particle diameters were calculated using
the volume of an ellipsoid for nonspheri-
cal fungal particles and ignoring the possi-
ble surface roughness.
Estimation of Respiratory Deposition
In estimating respiratory deposition of
fungal particles, the measured size distri-
butions for fungal particles were com-
bined with respiratory deposition data. A
semiempirical model by Stahlhofen et al.
(1989) was selected for this analysis be-
cause the model combines experimental
data from several research groups. Size
distribution curves for fungal particles
were multiplied by the deposition effi-
ciency curves of particular lung regions
for particular breathing patterns. The ar-
eas under these combined curves were
calculated and multiplied by the respira-
tory flow rates giving the number of de-
posited fungal particles per hour.
Fungal particle number concentrations
were standardized with respect to the im-
pactor's size intervals by dividing the con-
centration for each of the six intervals
(cfu/m3) by interval width (A log d,, pm).
Cubic spline interpolation (Stoer and Bu-
lirsch, 1980) was used to obtain more in-
termediary values for number concentra-
tion. The particle size range 0.8-8.5 p m
was divided into 100 evenly spaced parti-
cle sizes, for which the fungal particle
number concentrations were interpolated.
Respiratory deposition coefficients were
calculated for the above described parti-
cle sizes from equations given by
Stahlhofen et al. (1989). The calculations
were done separately for nasal, laryngeal
(mouth, larynx and pharynx), tracheo-
bronchial and alveolar deposition. The
respiratory deposition coefficient for each
region was the number of particles de-
T. Reponen
posited in the region divided by the num-
ber of particles inhaled. The interpolated
fungal particle concentrations were multi-
plied by the corresponding coefficients of
respiratory deposition giving the number
of deposited particles as a function of
particle size. All deposited particles were
summarized giving the total number of
deposited fungal particles per hour (de-
termined from colony-forming units, cfu;
cfu/h) by the following equation:
where
Q = mean breathing rate (cm3/s),
R, = interpolated respiratory depo-
sition coefficient,
N, = standardized and interpolated
fungal particle number con-
centration, cfu/(m3 pm),
A log d,,, =interpolation interval size
( pm).
Calculations were performed for adult hu-
mans at three breathing patterns asso-
ciated with different activity levels
(Martonen et al., 1992). A respiratory flow
rate of 117 cm3/s and nasal breathing
were used to simulate the lowest activity
level, e.g., a sedentary person. A flow rate
of 336 cm3/s and nasal breathing were
used to simulate light activity, e.g., domes-
tic work. A flow rate of 500 cm3/s, still
regarded in light activity, was used to sim-
ulate the activity level when people switch
to oral breathing (Kleinman, 1984).
RESULTS AND DISCUSSION
Size Distributions of Individual Fungal
Genera
A previous analysis of these data (Re-
ponen et al., 1994) showed that total con-
centrations of viable fungal particles were
Fungal Particle Size and Respiratory Deposition
higher in mold problem versus reference
dwellings. The concentration difference
was clearest in the size range 2.1-3.3 pm,
i.e., in the most common range for fungal
particles. In addition to concentration
differences, geometric mean of fungal
particle diameter was larger in mold prob-
lem than reference dwellings. In fall, rela-
tive humidity was higher in mold problem
(50%) than reference dwellings (42%),
whereas in winter, humidity level was the
same in both groups of dwellings (42%).
No apparent explanation for the differ-
ences in size distributions of fungal parti-
cles could be found in a general analysis
of the data.
Altogether 26 fungal genera were re-
covered from the air samples. However,
the frequency of occurrence for most gen-
era was so low, that their size distribu-
tions could not be calculated. Therefore,
only those fungal genera or groups pre-
sent in 2 50% of samples, i.e., Penicil-
lium, Cladosporium, and Aspergillus spp.,
and yeasts, were included in the detailed
analysis. In addition, d,,,,, values were
calculated for Oidiodendron spp., the con-
centration of which was higher in mold
problem buildings, and for Acremonium,
Botvyosporium , Oedocephalum, and
Stachybotrys spp., which were found only
in mold problem dwellings in winter (Re-
ponen et al., 1994).
The most common fungal genera or
groups had maximum concentrations in
the size range 2.1-3.3 p m (Figures 1-4).
Thus, the peaks in the size distributions
for total fungal particle concentrations re-
AERODYNAMIC DIAMETER (urn)
flected the combined effects of several
genera. The gaps in the size distributions
in Figures 1-4 may be caused by the
distinct and quite sharp size distributions
of different species within each genus.
Fungal particle concentration in mold
problem dwellings was higher than in ref-
erence dwellings in every size class for
Penicillium and Aspergillus spp. both in
fall and winter (Figures 1 and 3). In fall,
Penicillium spp. spore concentration was
higher indoors than outdoors in both
groups of dwellings. The size distribution
of fungal spores in indoor air also differed
from that in outdoor air. The size distri-
bution of Aspergillus spp. spores in out-
door air is not presented because this
genus occurred in only one of 12 outdoor
samples. These findings support previous
studies where Penicillium and Aspergillus
spp. were shown to originate mainly from
indoor sources (Fradkin et al., 1987; Flan-
nigan et al., 1991; Pasanen et al., 1991b).
The concentration of Cladosporium
spores was lower in indoor than outdoor
air in fall but the size distributions in-
doors and outdoors were similar indicat-
ing that the main source for Cladosporium
spp. spores was outdoor air (Figure 2).
However, in winter, the concentration of
Cladosporium spp. spores was higher in
mold problem than reference dwellings in
the size range 2.1-4.7 pm, indicating un-
usual indoor sources for Cladosporium in
this size range.
For yeasts, no clear differences be-
tween mold problem and reference
dwellings were seen in fungal particle
FIGURE 1. Size distribution of Penicillium spp.
spores in mold problem (--) and refercncc
(- - -) dwellings. The distribution is the geometric
mean for samples in following groups: A = Fall
1990, outdoor ( n = 6); B = Fall 1990, indoor
( n = 14); C = Winter 1991, indoor ( n = 12).

1 10 0.3 1 10 0.3 1 10
AERODYNAMIC DIAMETER i,um)
FIGURE 2. Size distribution of Cladosporium
spp. spores in mold problem (--) and refer-
ence (- - -) dwellings. The distribution is the geo-
metric mean for samples in following groups:
A = Fall 1990, outdoor ( n = 6); B = Fall 1990,
indoor ( n = 14); C = Winter 1991, indoor ( n =
12).
concentration or size distribution (Figure
41, indicating that the main sources for
yeasts were outdoors.
The effect of outdoor air infiltration on
indoor fungal concentration was seen
clearly both in particle size distribution
curves and d, values for Cladosporium
spp. and yeasts (Figures 2 and 4, Table 1).
The size distribution of these fungi in
indoor air shifted towards smaller parti-
cles supporting Raunemaa et a l . ' ~(1989)
earlier finding that fewer larger particles
infiltrate from outdoor into indoor air.
Measured d, values for each fungal
genus showed a large variation but d,,,,,
values ranged predominantly from 2-3
p m (Table 1). The d,,,,, values were close
to those obtained in laboratory experi-
ments at low humidities ( 5 40%) (Pasanen
et al., 1991; Madelin and Johnson, 1992)
but were remarkably smaller than the cal-
culated d, presented in Table 2. Possible
explanations for this are swelling of fungal
particles during the staining procedure
used for microscopic examination, or that
naturally occurring fungal particles may
be less than unit density (Table 3).
Calculated particle sizes for aggregated
fungal particles (Table 3) agreed well with
T. Reponen
AERODYNAMIC DIAMETER (,urn)
FIGURE 3. Size distribution of Aspergil . -
spores in mold problem (--) and reference
(- - -) dwellings. The distribution is the geometric
mean for samples in following groups: A = Fall
1990, indoor ( n = 14); B = Wintcr 1991, indoor
( n = 12).
u - -
0.3 1 10 03 1 10 0.3 1 10
AERODYNAMIC DIAMETER (,urn)
FIGURE 4. Size distribution of yeasts particles
in mold problem (--) and reference (- --)
dwellings. The distribution is the geometric mean
for samples in following groups: A = Fall 1990,
outdoor ( n = 6); B = Fall 1990, indoor ( n = 14);
C = Wintcr 1991, indoor ( n = 12).
Lacey's (1991) experimental results. In
Lacey's study, the d, for a chain of eight
fungal particles was only ca. 50% larger
than a single fungal particle while d, for a
compact aggregate of eight fungal parti-
cles was ca. 80% larger.
Measured d, values differed statisti-
cally significantly among the most com-
mon fungal genera or groups, Penicillium,
Cladosporium, Aspergillus, and yeasts
( p = 0.001, Friedman test). Aspergillus
spp. had the lowest d, values and Cla-
dosponum spp. and yeasts the highest (Ta-
Fungal Particle Size and Respiratory Deposition 17

TABLE 1. Measured Aerodynamic Particle Sizes ( pm) of Common Fungal Genera or Groups in
Mold Problem and Reference Dwellings."
Mold Problem Reference
Sampling Period/Genus n ' g , avg GSD n ' g , avg GSD
Full, indoor
Total
Penicillium
Cladosponum
Aspergillus
yeastsh
Oidiodendron
Fall, outdoor
Total
Penicillium
Cludosporium
Yeasts
Winter, indoor
Total
Penicillium
Cludosponum
Aspergillus
Yeasts
' n = number of posilive samples out of 14 samples indoors in fall, 6 samples outdoors in fall and 12 samples indoors in
winter; dg,avg = average geometric mean diameters of n samples; GSD = gcometric standard deviation of d,,,,,.
"d,,,,, statistically significantly higher ( p < 0.05) in mold problems vcrsus reference dwellings (Wilcoxon test).

ble 1). The d, values within each genus
did not vary statistically significantly be-
tween the fall and winter periods, suggest-
ing that fungal particle release mecha-
nisms did not change much with season.
The d,,,,, for total fungal particles was
larger in mold problem than reference
dwellings (Reponen et al., 1994). A similar
difference was seen also in fall for As-
pergillus, and Oidiodendron spp. and yeasts
(Table 1). However, d,,,,, for yeasts was
larger in outdoor air samples at mold
problem dwellings than reference dwel-
lings, perhaps due to differences in yeast
species at the various sites. However, this
proposed explanation cannot be con-
firmed from these data because yeasts
were not identified to species level.
The large size of Aspergillus and Oidio-
dendron spp. in mold problem dwellings
was probably caused by the higher relative
humidity in these buildings. This differ-
ence was seen only in fall when relative
humidity was higher in mold problem than
reference dwellings (Reponen et al., 1994).
In winter, relative humidity did not differ
between the two sets of buildings and only
yeasts showed different d, values. Other
possible explanations for the observed
differences in d, values could be different
distributions of fungal species, variations
in fungal particle age or degree of aggre-
gation. However, were these explanations
accurate, the effects of these differences
would have been expected also in winter
which was not the case.
Besides the most common fungal gen-
era described above, some rare fungal
genera also affected d, in mold problem
dwellings. Acremonium, Botryosporium,
Oedocephalum, and Stachybotys spp. were
found only in mold problem dwellings in
winter and, more frequently in mold prob-
lem dwellings in fall. The corresponding
dg ,avg values were 2.4, 2.6, 6.2, and
4.2 pm, respectively. Thus, Oedocephalum
18 T. Reponen

TABLE 2. Physical and Aerodynamic Diameters (d,, pm) of Indoor Airborne Fungal Particles, d,
Calculated Assuming Particle Density = 1 g/cm3.
Fungal Physical Measured
Species Sizea% d, Calculated d ,
Common in zndoor
C. cladosponoides (3-7) X (2-4) 2.3-2.5' 2.3-4.8
C. herbarum (5.5-13) X (4-6) - 4.4-7.6
C. sphuerospemum 3-4.5 - 3.0-4.5
P. breuicompactum 3-4.5 - 3.0-4.5
P. chrysogenurn (3-4) X (2.8-3.8) 2.6-3" 2.9-3.9
P. glabrum 3-3.5 - 3.0-3.5
P. uemcosum 3-4 - 3.0-4.0
A. penicilloides (3-3.5) X 4.5 - 3.4-3.8
A. niger 3.5-5 - 3.5-5.0
A . uersicolor 2-3.5 - 2.0-3.5
Indicators of indoor mo1dproblems:f
A. fumigatus 2.5-3 1.9-3.1"~"~ 2.5-3.0
Stachybotrys atra (7-12) X (4-6) 4.6' 4.7-7.4
Trichoderma uiride (3.5-4.5) X (3.6-4.8) - 3.5-4.6
Rhodotomla spp. (2.5-5) X (3-13) - 2.6-4.5
%amson and Reenen-Hockstra, 1988.
ell e t al., 1984.
'Fradkin e t al., 1987.
d ~ e r h o e f fe t al., 1990.
eMadelin and Johnson, 1992 (geomctric mean acrodynarnic diameter).
f ~ a m s o ne t al., 1993.
gLacey and Dutkiewitz, 1976 (calculated from sedimentation ralc).
h ~ a s a n e nc t al., 1992 (geometric mean aerodynamic diameter).
'Sorenson c t al., 1987 (mass median aerodynamic diamctcr).

and Stachybotrys spp. partly increased the
d, values for total fungal particles in mold
problem dwellings.
The measured fungal concentrations
and size distributions seen in this study
apply to a subarctic climate where build-
ing construction is usually tight decreasing
infiltration of outdoor fungal particles into
indoor air. In addition, outdoor concen-
trations of fungal particles are low in win-
ter due to snow cover, which facilitates
identification of indoor sources. On the
other hand, fungal species regarded as
indicators of mold problems are fairly
similar around the world (Samson et al.,
1994).

TABLE 3. Effcct of Fungal Particle Aggregation and Density on d, ( pm) for P. chrysogenum with
Average Physical Diameter of 2.8 X 3 p m
Calculatcd d,,
Density = 1g/cm3 Density = 0.5 g/cm3
Single particle
4-particle chain
Compact 4-particle
aggregate
8-particle chain
Compact 8-particle
aggregate
Fungal Particle Size and Respiratory Deposition

Respiratory Deposition dependence on breathing pattern as re-

The observed differences in size distribu-
tions of fungal particles between mold
problem and reference dwellings would
result in differential deposition of fungal
particles in the human respiratory system.
In this study, a preliminary estimation of
fungal particle deposition into different
parts of the respiratory system was made.
The effect of hygroscopicity on fungal
particle size could not be taken into ac-
count because there is not yet sufficient
data to include this. If fungal particles
were hygroscopic, d, would increase and
respiratory deposition would change (Li
and Hopke, 1993). The calculations on
numbers of deposited fungal particles pre-
sented here apply to adult humans. Alveo-
lar deposition is lower in children's res-
piratory system, whereas tracheobron-
chial deposition higher than for adults'
(Martonen et al., 1989).
Figure 5 shows respiratory deposition's
ported earlier, e.g., by Vincent (1990). The
effect of breathing pattern on particle de-
position is presented for the total concen-
tration of viable fungal particles. The
highest total deposition was observed for
a light activity breathing pattern in combi-
nation with oral breathing and the lowest
deposition for sedentary activity in combi-
nation with nasal breathing. The numbers
of deposited fungal particles were higher
in mold problem than reference dwellings
reflecting concentration differences.
During nasal breathing, 30%-50% of
fungal particles deposited in the nose and
ca. 30%-40% in the alveoli, whereas dur-
ing oral breathing, when no nasal deposi-
tion occurs, ca. 70% of fungal particles
reached the alveolar area (Figure 5).
Bronchial deposition was ca. 20%-30%
and the laryngeal deposition only ca.
2%-10% of total deposition. These pro-
portions were similar for mold problem

A. Q = 17 CG~IS 6. Q = 336 c m 3 / s

FALL 1990

FALL 1990
FALL 1990 WINTER 199 WINTER 199 WINTER

FROB REF . FROB REF PROB REF . PR08 REF 'ROB REF .
B
PROB REF

FIGURE 5. Regional respiratory deposition of fungal particles

for three breathing patterns: A. respiratory flow rate = 117
cm3/s and nasal breathing (sedentary), B. respiratory flow
rate =336 c r n v s and nasal breathing (light activity), C. respira-
tory flow rate = 500 cm3/s and oral breathing (light activity).
Deposition values calculated from size distributions of the total
concentration of viable fungal particles from Reponen et al.
(1994). PROB = mold problem, REF = reference dwellings.
20 T. Reponen

and reference dwellings when calculated
from total concentration of fungal
particles.
The number of deposited fungal parti-
cles for individual genera is presented for
light activity breathing pattern with nasal
breathing (Figure 6). For Penicillium and
Cladosporium spp., the ratio of number of
deposited particles for mold problem and
reference dwellings was the same as the
ratio for the concentration values. In con-
trast, for yeasts, the number of deposited

A. PENlClLLlUM spp. 3. CLADOSPORIUM spp.

FALL 1 9 9 0

FALL 1990

WINTER 1 9 9 '

WINTER 1 9 9 '

PROB REF . PROB REF PROB REF , PROB REF

C. ASPERGILLUS spp. D. YEASTS

FALL 1 9 9 0

WINTER 1 9 9 '

WINTER 1 9 9 '

PROB REF . PROB REF PROB REF . PROB REF

FIGURE 6. Regional respiratory deposition of major fungal

genera or groups for respiratory flow rate = 336 cm3/s and nasal
breathing. Deposition calculated from size distributions shown
in Figures 1-4. Note the different scales in figurcs. PROB = mold
problem, REF = reference dwellings.
Fungal Particle Size and Respiratory Deposition
fungal particles was lower in mold prob-
lem than reference dwellings although the
concentration was 1.4 times higher. With
Aspergillus spp., the concentration ratio
between mold problem and reference
dwellings was only two, while the ratio
of number of deposited particles was six
for total deposition and nine for alveolar
deposition. Aspergillus is a remarkable
genus, including species that may be aller-
genic, toxigenic or pathogenic (Burge,
1989; Flannigan, 1991; Miller, 1992).
Proportional deposition for individual
fungal genera were similar in mold prob-
lem and reference dwellings with some
exceptions. Due to particle sizes, the pro-
portional deposition of Aspergillus spp. in
the alveolar area was higher in mold prob-
lem than reference dwellings. For yeasts,
the situation was reversed: the proportion
of alveolar deposition was lower whereas
the proportion of nasal deposition higher
in mold problem than reference dwellings.
CONCLUSIONS
This analysis of the size distribution of the
main fungal genera or groups supports
previous findings that the sources of Peni-
cillium and Aspergillus spp. are mainly
indoors both in mold problem and refer-
ence dwellings. Furthermore, fungal con-
centrations were in every size class higher
in mold problem dwellings indicating that
a part of these fungal particles originated
from unusual indoor sources. Cladospo-
rium spp. and yeasts originated mainly
from outdoor sources, but in winter in-
door sources in mold problem dwellings
contributed to Cladosporium spp. concen-
trations. An effect of building infiltration
was seen in the indoor and outdoor size
distributions of Cladosporium spp. and
yeasts, i.e., an exclusion of larger fungal
particles.
The size distribution for total fungal
particles in mold problem dwellings was
skewed towards larger particle sizes when
21
compared with reference dwellings. This
difference was presumed due in part to
the genera Aspergillus and Oidiodendron,
whose larger size in mold problem
dwellings was associated with higher hu-
midity, and in part to yeasts, whose larger
size was probably due to the differences
in distributions of yeast species. Also some
rare fungal genera with larger than aver-
age d,, e.g., Oedocephalum and Stachy-
botrys, also contributed to the larger d,
of fungal particles in mold problem
dwellings. This difference in fungal parti-
cle size distribution between mold prob-
lem and reference dwellings would be
expected to cause differences in the air-
borne behavior of fungal particles be-
cause the particle settling velocity is a
function of squared diameter. However,
the difference in size distribution of total
fungal particles between mold problem
and reference dwellings was too small to
cause differences in proportional particle
deposition in the adult human respiratory
tract. Nevertheless, differences were esti-
mated for individual fungal groups. As-
pergillus spp. spores would be expected to
deposit more in the alveoli of the occu-
pants of mold problem dwellings whereas
for yeasts the situation would be expected
to be reversed.
The most common fungal genera had
their maximum concentrations in the size
range 2.1-3.3 pm, which led to high pro-
portional deposition in the alveoli because
alveolar deposition for particles > 0.5 p m
also has a maximum in the size range 2-3
pm. From the respiratory deposition cal-
culations with the most obvious breathing
patterns . for the home environment,
30%-50% of fungal particles would be
expected to deposit in the nose and
30%-40% in the alveoli during nasal
breathing, whereas 70% would deposit in
the alveoli during oral breathing. The pro-
portion of alveolar deposition was highest
for Aspergillus spp. (the smallest fungal
particles) and lowest for yeasts (the largest
22
fungal particles). Alveolar deposition is a
concern because clearance of particles
from this region is much slower than from
other parts of the respiratory tract due to
the lack of ciliary action (Vincent, 1990).
The dose-response relationships for
fungal particles are understood at present
poorly, i.e., the number of particles of
each fungal genus or species needed to
cause a certain symptom or disease is not
known. Therefore, in this study, respira-
tory exposure in mold problem houses
was compared with that in reference
houses rather than considering only abso-
lute air concentrations or numbers of de-
posited particles. This approach indicated
a higher exposure estimate for Aspevgillus
spp. by calculating the number of de-
posited spores than would have been ex-
pected based solely on airborne spore
concentration.
The author thanks Prof. K. Willeke, University of
Cincinnati, Dr. J. Ruuskanen, University of Kuopio and
Dr. A. Nevalainen, National Public Health Institute of
Finland for their helpful comments on this paper.
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Received January 4, 1994; accepted March 28, 1994.