The Complexity of Damp Indoor Spaces

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The Complexity of Damp Indoor Spaces

• Jack D. Thrasher, Ph.D.
• Sandra Crawley, M.ED., LADC
• Michelle DeFazio, BSAT NTEF
• 1

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Introduction

• Water Intrusion fungi, bacteria and protozoa
• Multiple Biocontaminants
• Additive and Synergistic Effects
• Increased Health Risks
• 2

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Additive and Synergistic Effects

• It is apparent that additive and synergistic
  effects have been demonstrated in vitro and in
  vivo.
• Vomitoxin and LPS increase apoptosis in lymphoid
  organs and proinflammatory cytokines in murine
  models
• Roridin A and LPS potentiates neurotoxicity in
  nasal airways of mice
• Aflatoxin B1 and LPS synergistic toxicity to
  liver 3

4
References for Introduction and Public Health

• Anderson et al (1997), Appl Environ Microbiol,
  63387-93.
• Peltola et al (2001a,b) Appl Environ Microbiol
  673260-75 4293-4304
• Hirvonen et al (2005) Indoor Air
  25(suppl9)65-70.
• Rintala et al (2001) Lett Appl Microbiol
  23439-43.
• Fisk et al (2007) Indoor Air 17294-96.
• Mudarri et al (2007 Indoor Air 17226-35.
• Genuis SJ (2007) Eur J Intern Med 28516-23.
• Bernstein, et al J Allergy Clin Immunol
  9158591. 4

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References for Introduction Cont.

• LPS AND MYCOTOXIN SYNERGISM
• Islam Z et al (2002) Toxicol Appl Pharmacol
  28043-55.
• Islam Z et al (2007) Toxicol Sci 98525-41.
• Islam Z, Pestka JJ (2006) Toxicol Appl Pharmacol
  2653-63.
• Luyendyk JP et al (2003, 2003). Toxicol Sci
  68220-5 72171-81
• Baron CC, et al (2001) Hepatology 3344-73
• 5

Toxicol Sci 98525-41.
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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

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INDOOR BIOCONTAMINANTS

• Indicator molds
• Bacteria- Gram () and (-)
• Particulates Large Small (lt2 micron)
• Mycotoxins
• Exotoxins
• 7

• MVOCs
• Enzymes (Allergens)
• Hemolysin Siderophores
• Galactomannans
• 1-3 beta-Glucans
• Endotoxins

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Molds in Relation to Water Activity

• 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 lt0.85) 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 gt90) 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).
• 8

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Indicator Molds

• Aspergillus fumigatus, versicolor, flavus, niger,
  sydowii, amstelodomi
• Penicillium brevicompactum, chrysogenum,
  chrysogenum
• Trichoderma sp.
• Fusarium moniforme.
• Epiccocum sp.
• Stachybotrys chartarum (two chemotypes)
• 9

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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

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Particulates Indoor Air

• Two Groups of Particulates
• Spores and Hyphae at 2 microns and up.
• Fragments lt2 microns.
• 11

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Fine Particulates

• Gorny (2004) Filamentous microorganisms and their
  fragments in the Indoor Air A review. Ann
  Agric Environ Med 11185-97.
• Gorny et al (2002) Fungal Fragments as Indoor Air
  Biocontaminants. Appl Environ Microbiol 3522-31.
• Gorny et al (2003) Release of Streptomyces albus
  propagules from contaminated surfaces. Environ
  Res 9135-33.
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Summary of Gornys Work

• Particulates lt 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.
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Fine Particulates and Trichothecenes

• Brasel, et al (2005) Detection of airborne
  Stachybotrys chartarum macrocyclic trichothecene
  mycotoxins in the indoor environments. Appl
  Environ Microbiol 717376-88.
• Brasel, et al (2005) Detection of airborne
  Stachybotrys chartarum macrocyclic trichothecene
  mycotoxins on particulates smaller than conidia.
  Appl Environ Microbiol 71114-22.
• Brasel, et al (2004) Detection of trichothecene
  mycotoxins in sera from individuals in
  Stachybotrys chartarum indoor environments. Arch
  Environ Health 59317-23.
• Gottschalk C, et al (2008) Detection of
  Satratoxin G and H in indoor air from a
  water-damaged building. Mycopathologia
  166103-07.
• 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 45682-91 14

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Summary of Brasel and Van Emon Papers

• Homes contaminated with Stachybotrys and multiple
  health complaints. Tested for Trichothecenes in
  contaminated vs non-contaminated buildings and
  outdoors.
• Multistage filtration 5.0, 1.2 O.4 microns,
  SEM, HPLC, ELISA.
• Buildings contained from lt10 to gt1,300 ug/m3.
  Difference between experimental and control
  building, p lt0.001
• Sera testes n 44 (contaminated n 26
  (uncontaminated). GC/MS. The exposed had
  significantly higher concentrations in sera vs
  controls (p lt0.05gt0.001)
• Stachylysin sera of rat pup with nasal
  installation and 5 adult males in a contaminated
  environment. Stachylysin averaged 371 mg/ml of
  serum.
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Indoor Mycotoxins Cont.

• Smoragiewicz, et al (1993) Trichothecene
  mycotoxins in the dust of ventilation systems in
  office buildings. Int Arch Occup Environ Health
  65113-7.
• Engelhart, et al (2002) Occurrence of toxigenic
  Aspergillus versicolor isolates and
  sterigmatocystin in carpet dust from a damp
  building. Appl Environ Microbiol 683886-90.
• Chapin-Kadouch et al (2006)Mycotoins
  identification in moldy dwellings. J Appl Toxicol
  26475-9
• 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 734211-17.
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Ergosterol and Respiratory Illness

• Park et al (2008) Hydrophilic fungi and
  ergosterol associated with respiratory Illness.
  Environ Health Perspec 11645-50.
• 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.
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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
  436120-2.
• Lewis et al (2005) Detection of gliotoxin in
  experimental and human aspergillosis. Infect
  Immunity 73635-7.
• 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).
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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.

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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 1142883-9
• 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 731630-4
• Gorny, Dutkiewcz (2002) Bacterial and fungal
  aerosols in indoor environment in Central and
  Easter European countries. Ann Agric environ
  Med. 917-23
• Gorny (2004) Filamentous microorganisms and their
  fragments in the Indoor Air A review. Ann
  Agric Environ Med 11185-97
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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.
• CC phenotypes react to low concentrations, while
  TT carriers react to higher concentrations of
  endotoxins Wheeze not Asthma
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CD14 and TLR4 - Endotoxins

• Martinez (2007) CD14, endotoxin and asthma. Proc
  Amer Thoracic Soc. 4a221-5
• Martinez (2007) Gene-environment interactions in
  asthma. Proc Amer Thoracic Soc 426-31.
• Simpson et al (2006) Endotoxin exposure, CD14,
  and allergic diseases. Am J Respir Crit Care Med
  174386-92
• 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 601482-92.
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Actinomycetes

• Streptomyces Up to 13 different species
• Nocardia, at least two species
• Mycobacterium Several different species.
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Streptomyces and ToxicityThree Key Papers

• Huttunen et al (2004) Synergistic interaction in
  simultaneous exposure to Streptomyces
  californicus and Stachybotrys chartarum. Environ
  Health Perspec 112659-65.
• Andersson et al (1998) Mitochondrial poison by
  Streptomyces griseus strains isolated from an
  indoor environment is Valinomycin. Appl Environ
  Microbiol 124763-73
• Jussila, et al (2003) Systemic immune responses
  in mice after repeated exposure of lungs to
  spores of Streptomyces californicus. Clin Diag
  Lab Immunol 1030-7
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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.
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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 674293-304.
• 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

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Mycobacterium Non-Tuberculin

• Torvinen et al (2000) Mycobacteria and fungi in
  moisture-damaged building materials. Appl Environ
  Microbiol 726822-4
• 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
  110119-25
• 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 256S1-S55. 27

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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 TNF-alpha and Nitric Oxide
  synthase, mononuclear cells infiltration and
  increased LDH.
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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
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Other Irritants and Toxins

• 1,3-Beta glucans
• Galactomannans
• MVOCs
• Time does not allow us to go into full details
  of these contaminants.
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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 55383-91.
• 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
  1141099-1107.
• 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 98526-41
• 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
  226192-8.
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Air Pollution and Olfactory Tract- Humans

• Calderon-Garciduenas (2004) Brain inflammation
  and Alzheimers-like pathology in individuals
  exposed to severe air pollution. Toxicol Pathol
  42650-8.
• 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 36289-320.
• Calderon-Garciduenas et al (In press) Air
  pollution cognitive deficits and brain
  abnormalities a pilot study with children and
  dogs. Brain and Cognition
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Conclusions

• 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.
• The interactions of the by-products are complex
  and probably include synergism
• The biocontaminants of the indoor environment
  should be systematically tested for mold species,
  bacterial species as well as mycotoxins.
• Air, bulk, wipe and wall cavity samples with
  PCR-DNA analysis of mold and bacterial species
• 33