EMLAB P&K - Air-Fungi 2003
EMLAB P&K - Air-Fungi 2003
EMLAB P&K - Air-Fungi 2003
INTERPRETATION GUIDE
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INTRODUCTION
The goal of this guide is to provide our clients background information and testing support to assist in the
location of potential sources of indoor air quality problems associated with aerosols and bioaerosols such as
mold spores, fiberglass fibers, and shedding of building components.
Although it is often not possible for the analyst to precisely identify the particle, or specific emission source
by analyzing the air sample alone, alerting a client when an “atypical” concentration of a specific particle
category is present can help identify and locate a potential contamination source.
Identification and classification procedures require the use of both bright field (BF) and polarized Light
Microscopy (PLM). Analysis of biological constituents rely primarily on the use of bright field microscopy.
Analysis of inorganic constituents and fibrous materials requires the laboratory analyst to use both bright
field and Polarized Light Microscopy. With the addition of the Air-O-Cell CSI cassette to the line of sampling
products now in commercial use, additional analysis by Scanning Electron Microscopy and Energy
Dispersive X-ray analysis can now be performed. This expansion of capabilities now allows the analysis of
mineral and non-biologic particles that do not have definitive optical identification properties.
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THE EAA PARTICLE CLASSIFICATION SYSTEM
The EAA Particle Classification System uses a combination of morphology and optical properties to
classify particles. In some cases the classification may not accurately represent the exact identity of
an individual particle.
Fibrous glass fibers Fibrous transparent glass fibers (fiberglass & mineral
(Isotropic) wool) used primarily as insulation materials and fillers
in ceiling tiles.
Opaque particles Particles that are dark brown or black in color or are
optically opaque and appear black with transmitted
light.
Insect Parts All particle debris associated with insects including leg
parts, wing scales, and body fragments
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MOLD & FUNGI
Elevated mold spore concentrations in both the indoor and outdoor environment are
known to cause of allergy symptoms and occasionally responsible for respiratory illness in
immuno-compromised individuals. Elevated mold spore concentrations in the indoor
environment can be from outdoor infiltration; or from indoor growth sources when elevated
surface moisture and humidity exist for extended periods of time.
Mold and fungi require three basic criteria to colonize the inside of a building including:
- A source of moisture
- A food source
- Lack of surface disturbance and/or air movement
Moisture sources in buildings occur most commonly as water and/or sewer leaks,
moisture intrusion through walls and foundations, or as condensation around windows or
in HVAC systems. In some parts of country such as the Southeast U.S. for example, the
relative humidity during certain times of the year is high enough to act as a significant
moisture source alone.
Indoor food sources for mold can be any organic material provided by a flood or sewer
backup; or cellulosic materials present in the building such as carpet backing, linoleum
backing, drywall paper, ceiling panels, or the buildup of plant and/or skin cell fragments or
debris on inorganic surfaces. Skin cell fragments are a significant food and colonizing
source in office buildings and private homes where a high occupancy exists, or adequate
housekeeping is not performed.
Molds colonize most readily where air disturbance is minimal. For this reason, mold
colonization occurs most frequently in closed or concealed spaces such as closets,
storerooms, basements, refrigeration units; or on the back or underside surface of
furniture. 4
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MOLD & FUNGI
At present, it is generally accepted in the medical community that exposure to mold may result in
symptoms consistent with a cold, flu, allergy hay fever, or asthma in some people. Others have no
symptoms at all. It is also generally accepted that there are no long term or permanent health effects
from exposure to mold once the occupant is removed from the property. It is also generally
recognized in the medical community, that those who are known to be allergic to molds and those with
asthma mayhave a higher risk of allergic reactions and should take extra precautions when in such
situations.
Outdoor assemblages of mold spores are most commonly populated with over 90% of the following
spores (listed in approximate order of descending abundance):
- Cladosporium
- Mushroom-like fungi (Ascospores and Basidiospores)
- Alternaria
- Rusts and Smuts (colonizing primary flower and leaf parts)
- Aspergillus & Penicillium (soil and moist cellulosic surfaces).
All of the above mentioned spores colonize decaying vegetation and/or soil.
The most common molds susceptible to indoor amplification (over 90% of the typical mold growth) in
approximate order of descending abundance include:
- Penicillium
- Aspergillus (flavus, fumigatus, terrus, versicolor, niger)
- Cladosporium
- Stachybotrys
- Alternaria, Chaetomium
- Zygomycetes (Mucor & Rhizopus)
- Ulocladum, Trichoderma
- Basidiomycete fungi
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MOLD & FUNGI
When moisture intrusion becomes chronic and/or involves sewage contamination, tertiary mold growth such
as Stachybotrys, Chaetomium, and Ulocladum may become common along with increased concentrations of
bacteria. Chronic moisture can also initiate the colonization of wood destroying fungi. Over time, these
kinds of fungi will colonize and destroy structural wood components of a building and can result in very high
indoor airborne basidiospore concentrations.
A high variability in outdoor mold spore concentrations and distribution exists on a daily to hourly basis and
is dependent on local vegetation and micro-climate, the time of year, local weather patterns, and diurnal
variation. As a result, caution must be used when simultaneously comparing limited data sets of inside and
outside concentrations or over generalizing any set of data. Tables given below can serve as a guide to
evaluating the relative degree of indoor airborne mold spore amplification.
As a general rule, total indoor airborne spore concentrations in a typical clean HVAC supplied building are
less than the “average” regional outside concentrations, and/or less than approximately 1,500 cts/m3.
Aspergillus /Penicillium and other hyaline spores are on average less than 700 cts/m3. Indicator fungi such
as Stachybotrys, Chaetomium, Ulocladium are often recovered in low concentrations in indoor samples as a
result of normal infiltration, therefore, automatically assuming there is indoor growth when low
concentrations of any indicator species are detected is inappropriate. Remember, there is always a likely
exception to every rule or generalization, and because there is no direct relationship between
simultaneously collected indoor and outdoor samples, performing a direct comparison with limited sampling
is often be misleading. The range of expected variability (i.e. a factor of 5 to 10 fold differences) when
comparing limited data sets must also be considered.
Examples photos of common airborne mold spores found indoors are given below:
Spores commonly associated with infiltration (in approximate decreasing frequency of occurrence)
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POLLEN
The presence of pollen in the indoor environment is almost always the result of air infiltration from the
outdoor environment. In a typical HVAC supplied air building, airborne pollen concentrations will be
very low (less than 10ct/m3) or not detected at all. However, sensitive individuals can often mistakenly
attribute complaints to the interior of a building that are actually the result of exterior pollen or other
allergen sources. Often landscaping in building courtyards can also be a factor with perceived indoor
problems. The time of year and the individuals home environment and pathway to work may also be
significant factors to consider for potential exposure.
According to the literature, individual response to pollen is highly variable. Some individuals with pollen
allergies may begin to exhibit symptoms when airborne concentrations exceed approximately 50 cts/m3,
especially with grass or highly allergenic pollen such as ragweed. The time of day when symptoms are
pronounced is extremely critical for proper diagnosis. Because of the wide range and severity of pollen
allergenicity, consultation with an Allergist may be warranted in the rare occasions when significant
indoor pollen concentrations are encountered.
Pollen identification in the EAA analysis report are given as the genus when known, or as the taxonomic
classification (i.e. inaperturate, triporate, tricolpate, etc.) when the pollen cannot be readily identified.
•Note: All concentrations refer to measurements obtained during the growing seasons reflecting local
California data. Other regions could be higher or lower than approximate ranges given above.
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ALGAE AND FERN SPORES
When algal or fern spores (in wet climate areas) are detected in any concentration in "indoor" samples, a
stagnant water source is likely present somewhere in proximity to the air intake stream. Although
significant information is not readily available regarding health effects, algal are potential indicators of
persistent moisture and other potential bacteriological or protozoa reservoirs.
DESCRIPTION Cts/m3
Outside air (depending on micro-climate -- 10 - 1,000
Inside air "clean Southern California building ND - 50
Inside air “moderate" 50 - 200
Inside air "high " > 200
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SKIN CELL FRAGMENTS
The skin cell fragment category includes skin cell fragment concentrations 20.0um in diameter. Dander or
skin cell fragments are the most common and major source of particle debris in indoor samples. One of
the biggest differences between "inside" and "outside" air quality is the concentration of airborne skin cell
fragment concentrations, and human-borne contaminants (i.e bacteria, viruses) riding as passengers on
skin tissue. Skin fragments often comprise over 50% of the "volume" of identifiable particles in indoor air.
It is not possible in a microscopic analysis to differentiate human dander from animal or pet dander.
Although no direct health effects can be derived by their measurement, skin cell fragment concentrations
are a good combined surrogate indicator of effective fresh air transfer rates, occupant density,
commensal bacteria potential, house-keeping and cleaning practices, and filtration of recirculated air in
the building. Typical concentrations encountered in buildings are given below.
DESCRIPTION Cts/m3
Outside air -- 50 - 1,000
Inside air "clean building 1,000 - 10,000
Inside air " high human activity" 10,000 - 20,000
Inside air "high personnel density and/or poor house-keeping" 20,000 - 100,000
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BIOLOGICAL, CELLULOSIC, & SYNTHETIC FIBERS
The cellulosic / synthetic fiber category covers a wide range of fibers that are commonly found in indoor
samples. Airborne fibers in this category include biogenic (derived from biological activity) fibers,
cellulosic fibers (derived from plants), and common synthetic fibers such as rayon, nylon, etc. Indoor
fiber emission sources include architectural finishes, paper products, clothing, and carpeting, and are
commonly found in airborne concentrations ranging from 100 to 1,000 cts/m3. These fibers are for the
most part are “anisotropic”, and produce a positive sign of elongation and will appear yellow when
oriented nw / se, and blue when oriented ne / sw in the PLM microscope. Some synthetic fibers and
monoclinic crystals appear yellow in both vibration directions, i.e., the sample light vibration in all
directions. Biogenic fibers (shedding from biological sources (plant, insect, or animal) by themselves
are not normally a cause of allergy or illness symptoms. Elevated biogenic fibers may be an indication
of inadequate housekeeping ventilation, high biogenic sources, and or high occupancy rates.
DESCRIPTION Cts/m3
• Outside air -- (Usually plant fragments) 100 - 1,000
• Inside air "clean buildings” (Usually fabric and paper fragments) 100 - 1,000
• Inside air " high human activity" 1,000 -5,000
• Inside air "high personnel density and/or poor house-keeping" 5,000 - 50,000
Cardboard box (BF) (PLM / full wave) Nylon carpet (BF) (PLM / full wave)
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FIBERGLASS FIBERS
Airborne fiberglass fibers encountered in the indoor environment are most commonly
composed of amorphous (non-crystalline) fibrous glass particles commonly known or
identified in products as fiberglass or mineral wool. Fibrous glass sources may
include ceiling tiles, debris from renovation projects, or the degradation of HVAC
system sound liners. More importantly, airborne detection is a strong indicator of
concurrent surface contamination and the potential for skin or eye contact irritation.
Because “fiberglass” and “mineral wool” are manufactured by different processes,
they are morphologically different. Fiberglass fibers are uniform along the entire
width of the fiber, while “mineral wool” is characterized by non-uniform width and the
presence of bulbous and rounded ends. Both categories of fibers are “isotropic” and
by definition the refractive index does not change with orientation. As a result,
fiberglass fibers when viewed in cross-polarized light become invisible without the use
of a retardation wave plate with the polarized light microscope. When a full wave
retardation plate is inserted, these fibers will appear colorless in all orientations. The
macroscopic coloration is due to the resin binder and not the color of the fiber.
DESCRIPTION Cts/m3
Outside air -- ND – 20
Inside air “low 20 - 50
Inside air " moderate“ 20 - 100
Inside air "high – building shedding likely" > 100
Yellow duct wrap insulation Mineral wool – ceiling tile Fiberglass (PLM-dispersion staining)
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OPAQUE PARTICLES
The opaque particle category encompasses a wide range of unrelated optically opaque particles
including combustion emissions (primarily diesel), paint and binders from degrading sound liners
in HVAC systems, biogenic debris (biological origin; i.e. insect droppings, decayed biological
debris, etc.), and rust from HVAC components, rubber tire particles, and copier and printer toner.
Specific identification of particle type usually requires additional sampling and analysis by
Scanning Electron Microscopy (SEM). This category of particle does not normally occur in
concentrations exceeding approximately 5,000 cts/m3 in "clean" indoor environments unless an
infiltration source (such as close proximity to roads or agriculture) is present. Identification of the
particle origin is not always possible, however, should be investigated as a potential contributor to
air quality complaints when airborne concentrations exceed 10,000 cts/m3. Commonly occurring
optically black / brown or opaque debris are generated from 3 general processes including
biogenic decay, corrosion, and combustion. Biogenic opaque black or brown debris are
generated from chemical, biological, or heat decomposition.
The most common outdoor sources are soil, vegetation, and automobile emissions. The most
common indoor biogenic sources (degradation from biological decay) are decayed dander, plant
fragments, and insect droppings. The most common non-biogenic sources are from the
degradation of metal HVAC system components (rust/metal fragments), binders and coatings
from insulation and architectural finishes, and combustion or heat generated degradation.
From a morphological standpoint, biologically derived opaque particles can often be separated
from other types of opaque particles. In some cases opaque particles cannot be morphologically
differentiated from corrosion shedding particles without using additional analysis by Scanning
Electron Microscopy or chemical analysis.
DESCRIPTION Cts/m3
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OPAQUE PARTICLES (Primarily biogenic)
Biogenic opaque black or brown debris are derived from the chemical or biological decomposition of
organically derived debris. The most common indoor sources are dander, plant fragments, insect droppings,
etc. From a morphological standpoint, biologically derived opaque particles can often be separated from
other types of opaque particles. Most biogenic debris have irregular, rounded, and “fuzzy” edge definition
and lack the presence of straight particle edges, cleavage planes, or fracture marks. They also have a
variability in optical density and will show an irregular variation in color and/or light transmission near the
edge of the particle, as well as throughout the particle. Examples of high levels of biogenic derived debris
(i.e. >100,000 cts/m3) are below:
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OPAQUE PARTICLES (Non-biogenic)
Non-biogenic black or brown debris are derived from the chemical or physical degradation,
corrosion, and shedding of mineral, resinous or film forming debris. The most common indoor
sources are drip pan scale, rust, or pigments from coated metals or insulation. From a
morphological standpoint, these types of opaque particles can often be separated from other
sources of opaque particles.
- Most non-biogenic opaque debris have well defined, angular, and distinct edge
definition and usuall have straight particle edges, cleavage planes, or fractures.
- Most non-biogenic opaque particles have a low variation in optical density (within the same
particle thickness) and do not show a large gradational variation in color and/or light
transmission near the edge or center of the particle.
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OPAQUE PARTICLES (Combustion debris)
Opaque combustion debris are derived from the heating burning of cellulosic materials, hydrocarbons,
or plastic residues. Angular and plate-like “Char” and “ash” particles are generated by the combustion
of plant material from forest fires. The most common indoor sources include the infiltration of outdoor
wood smoke or indoor fires. From a morphological standpoint, these types of opaque particles can be
separated from other types of opaque particles based on their shape, morphology, and residual plant-
like structure.
The combustion of fuel based organic material plastics, and plant resins produce small grape-like
spheroidal particle chains with individual particles at the limit of resolution of the optical microscope.
When these types of particles are “fresh”, positive identification is more easily achieved. When
combusted cellulose and aciniform particles are subjected to weathering and biodegration, it
becomes more difficult to differentiate them from other “opaque” debris.
It is important to note, emissions from indoor fires are derived from a combination of cellulose
sources, plastics, organic fuels, chemical compounds, and inorganic compounds. Indoor fires will
produce a combination of cellulosic combustion particles and well as “aciniform” particles associated
with hydrocarbon “combustion”. In addition, automobile emissions, especially diesel, produce
“aciniform” particles.
Opaque debris derived from soil infiltration in partially enclosed areas such as roll-up door storage
areas, warehouses, and crawl spaces. From a morphological standpoint, these types of opaque
particles cannot be readily separated from other types of opaque particles .
Warehouse next to an agricultural field – Opaque particle concentrations 150,000 ct/m3) – 800X
.
Mixture of rust & opaque particles (and mold spores) in construction debris – 800X
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OTHER PARTICLES – Insect parts
Most of the “other” biogenic particles include insect parts (i.e. body parts, antennae, legs, scales,
body hairs, and wing fragments. In "clean" indoor environments, airborne insect parts are
routinely detected, however, moderate concentrations above ~100 cts/m3 are not routinely
measured. Quantities of wings scales, body parts, or droppings (>500 cts/m3) may be an
indicator of an infestation or simply inadequate building maintenance and/or air filtration.
Occasionally dust mites are found in air samples when inadequate housekeeping or extensive
mold growth is present. The presence of dust or carpet mites in airborne samples may indicative
of an infestation.
DESCRIPTION Cts/m3
Outside air -- ND – 500
Inside air “low < 50
Inside air " moderate“ 50 -500
Inside air "high – building shedding likely" > 500
Dust & carpet mites Moth wing scale Moth wing scale
Elevated mineral construction debris particles are most commonly associated with renovation
activities. They are composed mostly of carbonate and gypsum dust generated from the
application and treatment of building components, drywall, patching compounds, and flooring
adhesives. In "clean" indoor environments, airborne concentrations are typically less than 5,000
cts/m3 and do not warrant routine measurement. However, when moderate concentrations are
observed in each field of view during the analysis they should be reported as a general comment
in regards to their presence and relative abundance (i.e. low, moderate, high).
DESCRIPTION Cts/m3
Outside air -- (expected range) ND – 1,000
Inside air “low <5,000
Inside air " moderate“ ~5,000 – 50,000
Inside air "high" > 50,000
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SUMMARY OF AIRBORNE PARTICLE CONCENTRATIONS
Outdoor Asp./Pen.
Indoor Asp./Pen.
Algae spores
Outdoor
Indoor
Pollen
Outdoor
Indoor
Outdoor Total
Indoor Total
Cellulose
Outdoor Total
Indoor Total
Opaque
Outdoor Total
Indoor Total
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