The Complexity of Damp
Indoor Spaces
Jack D. Thrasher, Ph.D.
Sandra Crawley, M.ED., LADC
Michelle DeFazio, BSAT – NTEF
#1
Introduction
• Water Intrusion – fungi, bacteria and
protozoa
• Multiple Biocontaminants
• Additive and Synergistic Effects
• Increased Health Risks
#2
Additive and Synergistic Effects
It is apparent that additive and synergistic
effects have been demonstrated in vitro
and in vivo.
1. Vomitoxin and LPS increase apoptosis in
lymphoid organs and proinflammatory
cytokines in murine models
2. Roridin A and LPS – potentiates
neurotoxicity in nasal airways of mice
3. Aflatoxin B1 and LPS – synergistic toxicity
to liver
#3
References for Introduction and Public Health
•
Anderson et al (1997), Appl Environ Microbiol, 63:387-93.
•
Peltola et al (2001a,b) Appl Environ Microbiol 67:3260-75; 4293-4304
•
Hirvonen et al (2005) Indoor Air 25(suppl9):65-70.
•
Rintala et al (2001) Lett Appl Microbiol 23:439-43.
•
Fisk et al (2007) Indoor Air 17:294-96.
•
Mudarri et al (2007: Indoor Air 17:226-35.
•
Genuis SJ (2007) Eur J Intern Med 28:516-23.
•
Bernstein, et al J Allergy Clin Immunol 91:585:91.
#4
References for Introduction Cont.
LPS AND MYCOTOXIN SYNERGISM
• Islam Z et al (2002) Toxicol Appl Pharmacol 280:43-55.
Toxicol Sci 98:525-41.
• Islam Z et al (2007) Toxicol Sci 98:525-41.
• Islam Z, Pestka JJ (2006) Toxicol Appl Pharmacol 26:53-63.
• Luyendyk JP et al (2003, 2003). Toxicol Sci 68:220-5; 72:171-81
• Baron CC, et al (2001) Hepatology 33:44-73
#5
Water Intrusion
Water Activity (Aw) – Water Content
The ratio of vapor pressure exerted
by water in the material to the vapor
pressure of pure water at the same
Temp. and Pressure
#6
INDOOR BIOCONTAMINANTS
• Indicator molds
• Bacteria- Gram (+)
and (-)
• Particulates: Large
& Small (<2 micron)
• Mycotoxins
• Exotoxins
#7
• MVOCs
• Enzymes (Allergens)
• Hemolysin &
Siderophores
• Galactomannans
• 1-3 beta-Glucans
• Endotoxins
Molds in Relation to Water
Activity
•
•
•
•
#8
This table summarizes the key water activity (aw) and molds that grow
most readily as primary, secondary and tertiary colonizers (adapted
from Nielsen, 2003; Gorny, 2004).
Primary colonizers (aw <0.85): Alternaria citri, Aspergillus (Eurotium)
amstelodomi, Aspergillus candid\us, glaucus, niger, penicilloides, repens,
restrictus, versicolor; Paedilomyces varlotti, Penicillium aurantiogriseum,
brevicompactum, chrysogenum, commune, expansum, griseoflavum;
Wallemia sebi
Secondary colonizers (aw = 85-90): Cladosporium cladosporoides,
herbarum, sphaerosperumum; Mucor circinelloides; Rhizopus oryzae.
Tertiary colonizers (aw = >90): Alternaria alternata; Aspergillus
fumigatus; Epicoccum species; Exophiala species; Fusarium moniforme;
Mucor plumbeus; Phoma herbarum; Phiaosphora species; Trichoderma
species; Stachybotrys chartarum; Ulocladium consortiale; Rhodotorula
species, Sporobolomyces species; and Actinomyces (Actinobacteria).
Indicator Molds
• Aspergillus fumigatus, versicolor, flavus,
niger, sydowii, amstelodomi
• Penicillium brevicompactum, chrysogenum,
chrysogenum
• Trichoderma sp.
• Fusarium moniforme.
• Epiccocum sp.
• Stachybotrys chartarum (two chemotypes)
#9
Identification of Molds
•
•
•
•
•
PCR DNA to determine species
Viable air samples
Bulk samples
Wipe Samples
Wall Cavity Samples
• Compare species indoors vs outdoors. If
Aspergillus, Penicillium. Stachybotrys &
Actinomycete species are above outdoors Contamination Exists
#10
Particulates – Indoor Air
Two Groups of Particulates
1. Spores and Hyphae at 2
microns and up.
2. Fragments <2 microns.
#11
Fine Particulates
1.
Gorny (2004) Filamentous microorganisms and
their fragments in the Indoor Air: A review.
Ann Agric Environ Med 11:185-97.
2.
Gorny et al (2002) Fungal Fragments as Indoor Air
Biocontaminants. Appl Environ Microbiol 3522-31.
3.
Gorny et al (2003) Release of Streptomyces albus
propagules from contaminated surfaces. Environ
Res 91:35-33.
#12
Summary of Gorny’s Work
• Particulates < 2 microns are aerosolized as
fragments from mycelia and spores.
• 320 times more concentrated than spores.
• Low Air velocity and vibration cause the release
of these fragments.
• The vibrations of 1-20 hertz with 1 hertz most
effective – normal human activity.
• Fragments contain allergens, enzymes, glucans,
galactomannans, mycotoxins, etc.
#13
Fine Particulates and Trichothecenes
• Brasel, et al (2005) Detection of airborne Stachybotrys
chartarum macrocyclic trichothecene mycotoxins in the indoor
environments. Appl Environ Microbiol 71:7376-88.
• Brasel, et al (2005) Detection of airborne Stachybotrys chartarum
macrocyclic trichothecene mycotoxins on particulates smaller
than conidia. Appl Environ Microbiol 71:114-22.
• Brasel, et al (2004) Detection of trichothecene mycotoxins in
sera from individuals in Stachybotrys chartarum indoor
environments. Arch Environ Health 59:317-23.
• Gottschalk C, et al (2008) Detection of Satratoxin G and H in
indoor air from a water-damaged building. Mycopathologia
166:103-07.
• Van Emon, et al (200) ELISA measurement of stachylysin in
serum to quantify human exposure to the indoor mold
Stachybotrys chartarum. J Occup Environ Med 45:682-91 #14
Summary of Brasel and Van Emon Papers
• Homes contaminated with Stachybotrys and multiple health
complaints. Tested for Trichothecenes in contaminated vs noncontaminated buildings and outdoors.
• Multistage filtration: 5.0, 1.2 & O.4 microns, SEM, HPLC, ELISA.
• Buildings contained from <10 to >1,300 ug/m3. Difference
between experimental and control building, p <0.001
• Sera testes: n = 44 (contaminated; n = 26 (uncontaminated).
GC/MS. The exposed had significantly higher concentrations in
sera vs controls (p <0.05>0.001)
• Stachylysin: sera of rat pup with nasal installation and 5 adult
males in a contaminated environment. Stachylysin averaged
371 mg/ml of serum.
#15
Indoor Mycotoxins Cont.
• Smoragiewicz, et al (1993) Trichothecene mycotoxins in the
dust of ventilation systems in office buildings. Int Arch Occup
Environ Health 65:113-7.
• Engelhart, et al (2002) Occurrence of toxigenic Aspergillus
versicolor isolates and sterigmatocystin in carpet dust from a
damp building. Appl Environ Microbiol 68:3886-90.
• Chapin-Kadouch et al (2006)Mycotoins identification in moldy
dwellings. J Appl Toxicol 26:475-9
• Bloom et al (2007) Mass spectrometry-based strategy for direct
detection and quantification of some mycotoxins produced by
Stachybotrys chartarum and Aspergillus spp in indoor
environments. Appl Environ Microbiol 73:4211-17.
#16
Ergosterol and Respiratory Illness
• Park et al (2008) Hydrophilic fungi and ergosterol
associated with respiratory Illness. Environ Health
Perspec 116:45-50.
• n = 200 with 123 respiratory disease = asthma
Asthma was significantly associated with presence
of hydrophilic and Ergosterol.
Ergosterol is a sterol found in fungal walls.
#17
Aspergillosis and Gliotoxin
•
•
Lewis et al (2005) Frequency and species distribution of
gliotoxin producing Aspergillus isolates recovered from
patients at a tertiary-care cancer center. J Clin Microbiol
43:6120-2.
Lewis et al (2005) Detection of gliotoxin in experimental and
human aspergillosis. Infect Immunity 73:635-7.
1.
Gliotoxin was produced by isolates: fumigatus (93%); niger
(75%), terreus (25%) flavus (4%).
2.
Mice: Lungs (m = 3,975 ng/g; Sera (M = 36.9 ng/ml)
Human Sera: 2 of 11 patients without confirmed I.A. In
Confirmed I.A. patients 4 of 5 (range 166 to 785 ng/ml).
#18
Bacteria Gram Negative and Positive –
Examples – Gorny & Dukiewicz, 2002; Bouillard
et al, 2004.
Gram Negative
Enterobacteriaceae
Pseudomonas sp.
Proteus sp.
Actinobacter sp.
Alcaligenes faecalis
(aerobes and
anaerobes)
#19
Gram Positive
Micrococcus sp.
Bacillus sp
Streptococcus sp
Enterococcus sp.
Mycobacteria –
Streptomyces sp,
Nocardia sp.,
Mycobacterium sp.
Endotoxins - Indoor
• Chew et al (2006) Mold and endotoxin levels in the aftermath of
hurricane Katrina: A pilot project of homes in New Orleans.
Environ Health Perspec 114:2883-9
• Rao et al (2007) Characterization of airborne molds,
endotoxins, and glucans in the homes of New Orleans after
hurricanes Katrina and Rita. Appl Environ Microbiol 73:1630-4
• Gorny, Dutkiewcz (2002) Bacterial and fungal aerosols in
indoor environment in Central and Easter European countries.
Ann Agric environ Med. 9:17-23
• Gorny (2004) Filamentous microorganisms and their fragments
in the Indoor Air: A review. Ann Agric Environ Med 11:185-97
#20
Endotoxins – Indoor Air
1. Inhaled LPS are associated with inflammation of the lungs
involving TNF-alpha, and activation of C3.
2. There is complicated interaction between TLR4 receptors and
CD14 polymorphism – CD14-CC and CD14-TT including
carrier proteins M88 and TRAM
3. IgE interactions are not the norm. IL4, IL5, eosinophils and
neutrophils are involved and difficult to control.
4.
#21
CC phenotypes react to low concentrations, while TT carriers
react to higher concentrations of endotoxins – Wheeze not
Asthma
CD14 and TLR4 - Endotoxins
1.
Martinez (2007) CD14, endotoxin and asthma. Proc Amer
Thoracic Soc. 4a;221-5
2.
Martinez (2007) Gene-environment interactions in asthma.
Proc Amer Thoracic Soc 4:26-31.
3.
Simpson et al (2006) Endotoxin exposure, CD14, and allergic
diseases. Am J Respir Crit Care Med 174:386-92
4.
Sackesen et al (2005) The effect of polymorphism at the CD14
promoter and the TLR4 gene on asthma phenotypes in
Turkish children with asthma. Allergy 60:1482-92.
#22
Actinomycetes
• Streptomyces – Up to 13 different
species
• Nocardia, at least two species
• Mycobacterium – Several different
species.
#23
Streptomyces and Toxicity
Three Key Papers
• Huttunen et al (2004) Synergistic interaction in simultaneous
exposure to Streptomyces californicus and Stachybotrys
chartarum. Environ Health Perspec 112:659-65.
• Andersson et al (1998) Mitochondrial poison by Streptomyces
griseus strains isolated from an indoor environment is
Valinomycin. Appl Environ Microbiol 12:4763-73
• Jussila, et al (2003) Systemic immune responses in mice after
repeated exposure of lungs to spores of Streptomyces
californicus. Clin Diag Lab Immunol 10:30-7
#24
Summary of Streptomyces Papers
• Synergistic action occurs with trichothecenes and S.
californicus with respect to IL-4, TNF-alpha in vitro.
• Vanlinomycin – caused mitochondrial swelling,
inhibiting oxidative phosphorylation and motility –
boar sperm cells.
• Spores of S. californicus cause inflammation in the
lungs and systemic immunotoxicity in the spleen
and other lymphoid organs.
#25
Toxic Nocardia
Peltola et al (2004) Isolation of toxigenic Nocardia strains from
indoor environments and description of two new
Nocardiopsis species, N. exhalans sp. nov. and N.
umisdischolae sp. nov. Appl Environ Microbiol 67:4293-304.
1.
Methanol soluble toxin that depleted ATP sperm
cells.
2.
Destruction of Cell membrane of sperm cells.
3.
The toxic properties suggests good survival in the
indoor environment.
#26
Mycobacterium – Non-Tuberculin
• Torvinen et al (2000) Mycobacteria and fungi in moisturedamaged building materials. Appl Environ Microbiol 72:6822-4
• Jussila et al (2002) Mycobacterium terrae isolated form indoor
air of a moisture-damaged building induces biphasic
inflammatory response in mouse lungs. Environ Health
Perspec 110:119-25
• Falkinham (2003) Mycobacterial aerosols and respiratory
disease. CDC. Emerging Diseases. 9(No. 7) July 2003.
• American Thoracic Society. (1997) Diagnosis and treatment
causes by nontuberculous mycobacteria. Respir Crit Care Med
256:S1-S55.
#27
Concerns of Nontuberculin Mycobacteria
•
•
Torvinen – Nontuberculin mycobacteria are present in water-damaged
building materials.
American Thoracic Society – Nontuberculin Mycobacterium infections
are on the increase worldwide. The cause is unknown.
•
Falkinham – Mycobacterium terrae, immunogenum and avium have
been implicated in outbreaks of Hypersensitivity pneumonitis.
•
Jussila et al – Two phase inflammatory response to M. terrae.
– A. Initial response lasts 6 hours to 3 days- TNF alpha & IL-6 and
neutrophils and increased LDH.
– B, Second Phase last up 7-28 days – increase production of TNFalpha and Nitric Oxide synthase, mononuclear cells infiltration and
increased LDH.
#28
Hemolysins
• In Cleveland, Ohio, pulmonary hemosiderosis was initially
blamed on S. chartarum, Stachylysin and a siderophore
produced by S. chartarum.
• Vesper and Vesper recently published data on hemolysins
produced by Aspergillus, Penicillium and other molds isolated
from the Cleveland Homes. Advances Appl. Microbiol, V 55.
A. 12 species of Aspergillus
B. 11 species of Penicillium.
C. Others – Emericella, Memnoniella, Trichoderma
species
#29
Other Irritants and Toxins
• 1,3-Beta glucans
• Galactomannans
• MVOCs
Time does not allow us to go into full details
of these contaminants.
#30
Olfactory Tract and Mycotoxins
•
Larrson, Tjaive (2000) Intranasal instillation of Aflatoxin B1 in rats:
Bioactivation in the nasal mucosa and neuronal transport to the
olfactory bulb. Toxicol Sci 55:383-91.
• Islam et al (2006) Satratoxin G from the black mold Stachybotrys
chartarum evokes olfactory sensory neuron loss and inflammation in
the murine nose and brain. Environ Health Perspec 114:1099-1107.
• Islam et al (2007) Neurotoxicity and inflammation in the nasal airways
of mice exposed to the macrocyclic trichothecene mycotoxin roridin a:
kinetics and potentiation by bacterial polysaccharide coexposure.
Toxicol Sci 98:526-41
• Shwe TTT et al (2008) Changes in neurotransmitter levels and
proinflammatory cytokine mRNA expressions in the mice olfactory
bulb following nanoparticle exposure. Toxicol Appl Pharm 226:192-8.
#31
Air Pollution and Olfactory TractHumans
• Calderon-Garciduenas (2004) Brain inflammation and
Alzheimer’s-like pathology in individuals exposed to severe air
pollution. Toxicol Pathol 42:650-8.
• Calderon-Garciduenas (2008) Long-term air pollution exposure
is associated with neuroinflammation, innate immune
response, disruption of the blood-brain barrier, ultrafine
particulate deposition and accumulation of amyloid beta-42 and
alpha-synuclein in children and young adults. Toxicol Pathol
36:289-320.
• Calderon-Garciduenas et al (In press) Air pollution: cognitive
deficits and brain abnormalities: a pilot study with children and
dogs. Brain and Cognition
#32
Conclusions
1.
As we have seen here today, the indoor environment
resulting from microbial growth in response to water
intrusion is very complex involving molds, bacteria and their
by-products.
2.
The interactions of the by-products are complex and
probably include synergism
3.
The biocontaminants of the indoor environment should be
systematically tested for mold species, bacterial species as
well as mycotoxins.
4.
Air, bulk, wipe and wall cavity samples with PCR-DNA
analysis of mold and bacterial species
#33