Indoor Air Quality Webinar - EMLab P&K · 2013-02-05 · Indoor Air Quality Webinar Mold: • What it is • Health Effects • Sampling • Data Interpretation EMLab P&K Will begin

Indoor Air Quality WebinarMold:

• What it is

• Health Effects

• Sampling

• Data Interpretation

EMLab P&K

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Seminar Agenda – Day 1

Mold1. What is mold?2. Health effects associated with mold exposure3. Sampling and analytical methods4. Laboratory data interpretation


Macrocybe TitansCosta Rica, 16 pounds, three feet in diameter


• Molds: visible filamentous fungal colonies

• Moldy odors: volatile organic compounds produced by most fungi

• Toxins: produced by many different kinds of fungi, some of which would not be considered “molds.”

• All fungi are NOT considered molds.

• All molds ARE fungi.

A Few Fungal Terms


• Kingdom Fungi– Diverse group defined by filamentous

lifestyle; require complex carbon compounds for food

– Mushrooms, plant rusts and smuts, molds, yeasts, etc.

– 1.5 million species– Only ~70,000 fungi currently categorized

• Fungal lifestyles– Saprobes (decay dead plant matter)– Plant parasites– Animal parasites– Symbiotic relationships

• Reproduce sexually and asexually by spores

What Are Fungi?


ClassOrder

FamilyGenus

Species

Example: Genus StachybotrysSpecies chartarum

Fungal Classification


The Fungal Body / The Inside-Out Organism


Fungal Physiology

• Excrete enzymes into the environment• Enzymes aid in digestion of carbohydrates and proteins into soluble

units• Soluble sugars and amino acids are absorbed into the fungal cell• Cell uses these foods to produce energy. • Steps in the production of energy lead to the production of

secondary metabolites– Antibiotics (e.g., Penicillin)– Volatile organic compounds (e.g., benzene)– Alcohols (e.g., ethanol) – Acids (e.g., citric acid)– Mycotoxins (e.g., aflatoxin)– CO2

– H20


Emericella(ascospores)

Aspergillus(conidia)

Polymorphism in Fungi


mycelium

mycelium

Nuclear fusion

meiosis

Sexual spores

mitosis

Asexual spores Sexual

ascospores

Asexual

conidia

Fungal Lifecycles


Spores: Disseminators

• Dry, hydrophobic• Wind disseminated

– Penicillium

– Aspergillus

– Cladosporium

– Alternaria

– Etc.


Spores: Disseminators

• Wet, hydrophilic• Water and insect disseminated

– Fusarium

– Acremonium

– Stachybotrys

– Chaetomium

– Etc.


Fungi in Nature

• Ubiquitous - they’re everywhere• Play a vital role in the environment

as: – Decomposers

– Plant pathogens

– Animals pathogens (esp. insects)

– Produce allergens

• Always some in both indoor and outdoor air

• Many man-made uses– Antibiotics

– Food products

– Chemicals


Outdoor Aerosol


Outdoor Aerosol Exposure Patterns

• Spores are always present in the outdoor air.• Spore aerosol distributions vary with season and

with climate (e.g., geographically)


Impact of Outdoor Aerosols on Indoor Aerosols

• Except when snow covers the ground, outdoor aerosol concentrations generally exceed those indoors

• Unless active growth is occurring indoors, the general pattern of aerosols is the same indoors and out.

• Outdoor aerosols enter through openings (windows, doors, cracks) and on surfaces (including clothing) brought indoors.


Growth of Outdoor Aerosols on Indoor Substrates

• Requirements for growth– Water, complex carbon compounds

• Growth rate– Germination rate: Minutes to days

– Vegetative growth: hours to weeks

• Competition– Sugar: lots of competition: fast growth rate

– Cellulose: much less competition: moderate growth rate

– Lignin: very little competition: slow growth rate


Botrytis on Strawberries


Cladosporium Growing in Condensation


Penicillium on an Orange


White Rot


Soft-Rot Fungi• Degrade cellulose and hemicellulose• Early stages of wood decay• High moisture and increased nitrogen content• Wood affected by soft rot may appear wet, spongy, or pitted. • 300 species of known soft-rot fungi. • Cephalosporium, Acremonium, and Chaetomium

Wood Rot Fungi

Major Components of Wood• Cellulose 40-50%• Hemicellulose 25-40%• Lignin 20-35%• Fungal spores do not germinate

readily on wood that has a moisture content below the fiber saturation point, around 25-30%.


Brown-Rot Fungi• Degrading cellulose and hemicellulose• Lignin remains intact and appears as a brown, crumbly matrix• Less than 6% of all wood-decay fungi• Basidiomycota (the mushrooms)

Wood Rot Fungi

Major Components of Wood• Cellulose 40-50%• Hemicellulose 25-40%• Lignin 20-35%


White-Rot (Dry Rot) Fungi• Degrade all the major components of wood (cellulose, hemicellulose, and lignin).• Wood to become progressively more fragile over time.• Most are macro fungi that produce visible reproductive structures such as mushrooms.

Wood Rot Fungi

Major Components of Wood• Cellulose 40-50%• Hemicellulose 25-40%• Lignin 20-35%


Stachybotrys?

Lumberyard Mold


Ceratocystis / Ophiostoma Group: ‘Lumberyard Molds’

• Grows quickly on wet sap wood• Becomes dormant when dry• Almost always present on lumber. • Integrity of wood not damaged;

superficial on surfaces• Dissemination of sticky spores by

arthropods; not airborne• Toxicity or allergenicity unknown,

but little if any exposure occurs from this source


Information on other fungi is available online at: www.EMLab.com

Ceratocystis/Ophiostoma Group

Distribution Allergen Growth Indoors Characteristics: Growth/Culture

Ubiquitous; cosmopolitan. Approx. 56 species, both genera.

Not studied. Persons most likely to be affected would be lumberyard workers or carpenters.

Most homes built with lumber have areas of growth on wood framing inside walls.

Some species may grow on specialized laboratory media. Asexual forms grow on general fungal media.

Where Found Potential Opportunist or Pathogen

Industrial Uses Notes on Spore Trap Recognition

Commercial lumber, tree and plant pathogen.

Not reported to infect humans or animals. A connection between Ophiostoma and the human pathogen Sporothrixschenckii has been proposed but not confirmed.

None known. Not identifiable on spore trap slides. On rare occasions the spores of Gonatobotryum, which grows parasitically on Ceratocystis, are seen on spore trap samples (an indirect indication of the presence of this group of fungi).

Mode of Dissemination Potential Toxin Production Other Comments Notes on Tape Lift Recognition

Wet spore. Insects. None listed. Ophiostoma ulmi is the cause of Dutch Elm Disease.

Distinctive fruiting bodies with long necks are identifiable on tape lifts.

Ascomycetes with multiple asexual forms: Graphium, Leptographium, Sporothrix-like anamorphs, Chalara.


Penicillium Gorgonzola


Health Effects


Fungal Disease

• Disease related to airborne fungi remains controversial.

• Except for medical treatment, little new information is available.

• This presentation reviews – The diseases and

– Present traditional and new information on cause/effects relationships and the environmental aspects of fungal disease


Anecdotal evidence: word of mouth, case studies

Experimental evidence Epidemiological evidence

Logic

“proof”

Types of Evidence


Allergic Disease

• Diseases– Allergic rhinitis (hay fever)

– Asthma

– Hypersensitivity pneumonitis(allergic pneumonia)

– Eczema (atopic dermatitis?)


Allergic Disease

• Background– Antibody/antigen related

– Multiple exposures required

– Allergens/antigens are usually proteins

– Fungi produce these proteins

– Most disease-causing exposure occurs outdoors

– New allergies can develop from indoor fungal growth


Evidence for Allergic Reactions

• Asthma:– There is extensive evidence to

document that exposure to fungi can precipitate asthma attacks, and may be involved in the initial development of asthma in some circumstances.

• Childhood asthma studies

• Work related asthma studies




Logic

“proof”

Types of Evidence for Fungal-Induced Asthma


Allergic Disease

• Asthma attack related to fungi on mouth guard (Glass et al., 2007)– If mouth guard is used only at

school, the school environment could be blamed

– If mold is found in a building, the tendency is to blame the mold for symptoms

– Important to consider other possibilities, especially if only one person is affected.


Lower Respiratory Infections

• Lower respiratory illness:– There is epidemiological evidence

that children living in damp buildings have an increased incidence of lower respiratory infections.

– These effects are separate from asthma, and don’t depend on sensitivity to the fungi, or to family history of allergies.




Logic

“proof”

Evidence for Fungal-Associated Lower Respiratory Infection


Fungal Irritants

• Irritants (volatile organic compounds):– There is some evidence that these

agents cause mucous membrane (eye, nose) irritation, and may lead to headaches.

– Evidence so far is incomplete but there is no indication so far that these effects do not occur.

– Neurological effects probably due to perception of odors




Logic

“proof”

Evidence for Fungal-Associated Irritant Effects


Fungal Colonization

• Diseases– Allergic bronchopulmonary aspergillosis (mycosis)– Allergic Fungal Sinusitis

• Background– ABPA: fungus colonizes mucous in the lung; symptoms caused by immune

response to the fungus– AFS: Fungus colonizes mucous in sinuses; sinuses fill with fungal hyphae

and thick mucous produced in response to the fungal allergens– Risk depends on having chronic asthma or rhinitis– Only a few fungi are involved


Fungal Infections

• Diseases– Virulent infections

– Skin infections

– Opportunistic infections

• Background– Virulent: Infect everyone without

specific immunity (like the flu)

– Skin infections: Infect skin surfaces only (ringworm, dandruff, athlete’s foot)

– Opportunistic: Infect only those with damaged immune defenses


Fungal Infections

– All require living spores for disease

– Virulent agents do not grow indoors and disease depends on disturbance of outdoor reservoirs

– Skin agents are probably not airborne

– Opportunistic agents common in the environment


Virulent Infections

• Histoplasmosis (Histoplasma capsulatum)– Wet soil enriched with bird droppings

– Soil disturbance necessary for dissemination

– Endemic in the Mississippi valley

• Blastomycosis (Blastomyces dermatididis)– Similar to Histoplasma

• Coccidioidomycosis (Coccidioides immitis)– Dry desert soils

– Sandstorms


Skin Infections

• Microsporum, Epidermophyton and Trichophyton– Athlete’s foot

– Ringworm

– Jock itch


Opportunistic Infections

• Can be a serious issue for the immune compromised:– AIDS

– Organ transplants

– Chemotherapy

– Bone marrow transplants

• Not normally an issue for people with competent immune systems

• Well documented




Logic

“proof”

Types of Evidence for Opportunistic Infections


Fungal Toxins

• Diseases– Cancer– Immunosuppression– Neurological effects– Irritant effects

• Background– All of these effects are related to

ingestion of moldy food. – Cancer requires long term

exposure; exposure cumulative– Other effects are immediate;

exposures not cumulative– Inhalation very unlikely as an

exposure route


Fungus Mycotoxins Possible health effects

Alternaria alternata Tenuazoic acid Nephro, hepato toxic, hemorrhagic

Aspergillus flavus, A. parasiticus

Aflatoxins Mutagenic, carcinogenic, hepatotoxic

Aspergillusfumigatus

Fumitremorgens, gliotoxin

TremorgenicCytotoxic

Cladosporium sp. Epicladosporic acid Immunosuppressive

Fusariummoniliforme

Fumonisins Neuro, nephro & Hepatotoxic, carcinogen

Stachybotryschartarum (atra)

Satratoxins, verrucarins, roridins

Immunosuppressive, hemotoxic, hemorrhagic

Mycotoxins


Fungal Toxins – New Information

• New information – Hundreds of publications on

dietary mycotoxins, especially in underdeveloped countries.

– No new publications that provide evidence for airbornemycotoxin exposure.

– Stachybotrys conidia require air speeds 1,000 times higher than those that normally prevail in the indoor environment. (Tucker et al. 2007)


Fungal Toxins

• Toxic molds on the web– Many sites on the web claim that

scientists are underreporting the dangers of toxic mold.

– Position papers by major medical organizations are considered “biased”.

– Expert witnesses are considered biased because they are paid for presenting their opinions.


Fungal Toxins

• Comments– Medical groups should not rely on reviews

for position papers, only the published peer reviewed research literature.

– Courts should not decide scientific relevance of scientific literature. Position papers are consensus documents of experts in the field, and should be considered authoritative.

– Just because scientists are often defense witnesses does not mean they do not know the facts. Experts are hired because they do know the facts. These same experts would be used by the plaintiffs if the facts fit their cases.

– The fact remains that the exposure models are relevant and there is no good evidence for mycotoxin inhalation disease




Logic

“proof”

Evidence for Fungal-Associated MycotoxinEffects (Inhalation)


On the Other Hand

• A June 2007 article in the Journal of Nutrition showed a positive effect on the immune system due to consumption of white button mushrooms.

• Study done on mice.• Helps promote innate immunity

against tumors and viruses


References

• Kim et al., Indoor Air 2007; 17: 153–163• Pini et al.,2008• Tucker et al., Indoor Air 2007; 17: 153–163• Burr et al., Thorax 2007;62:767–772.• Fisk et al., Indoor Air 2007; 17: 284–296• Holck et al., 2007 Basic & Clinical Pharmacology & Toxicology 101, 455–

458,• Iverson et al., Eur J Clin Microbiol Infect Dis (2007) 26:879–886• Nucci & Anaissie, CLINICAL MICROBIOLOGY REVIEWS, Oct. 2007, p.

695–704• Wang et al., Pediatr Allergy Immunol 2007: 18: 441–447• Park et al., Environ Health Perspect 116:45–50 (2008). • Glass et al., Gen Dent. 2007 Sep-Oct;55(5):4a36-40 • Sautour et al., J Hosp Infect. 2007 Dec;67(4):367-73.


Why Sample for Biological Materials?

• Find out if there is fungal growth in the environment.

Note that this can sometimes be done by a field inspection as well -directly observing fungi or finding risk factors.

• Find out if people are being exposed to more or different kinds of fungi than would normally be experienced.

• Help identify building defects(dry rot,…)


Fundamental Approach to Sampling

• Use a hypothesis driven, technically valid approach:– Develop a hypothesis

– Try to prove the hypothesis is wrong

– Develop an investigation strategy to test your hypothesis


Investigative Strategies

• Visual/sensory inspection– Find water intrusion sites

– Find visible mold

– Locate source of moldy odors

• Surface or bulk sampling– Verify visible mold

– Check for invisible mold

• Air sampling– Verify that aerosols can be produced

– Verify that exposure is occurring


Appropriate Analytical Methods For Various Sample Types

Sampling method Analytic method

Tape sampling Microscopy

Swab sampling Culture (microscopy)

Bulk sampling Culture, microscopy

Dust sampling Culture, ELISA, PCR, GCMS, (microscopy)

Air sampling: inertial impaction on sticky surface

Microscopy

Air sampling: inertial impaction on culture plate

Culture

Liquid impingement Culture, PCR, GCMS

Filtration PCR, (culture, microscopy)


Ye ol’ Chain of Custody

• This is a legal document:– May show up in court– Needs to be accurate– Needs to communicate

credibility

• Be sure that:– Contact information is

accurate– All samples are accounted for– Desired analysis is indicated

(especially for swabs and samples often analyzed in multiple ways)

– Special requests are large and clear


Surface Sampling

• Tapes:– Very good for most surfaces– Very poor for cultures– Poor results on wet surfaces– Try to leave a “tab” or “handle” for the lab

• Bulks:– Keep small (2” x 2”)– Can do direct exam (generally recommended)– Good for cultures

• Swabs:– Good for places that are hard to reach– Last choice for direct exam (try tape or bulk first)– Good for cultures– Ship for overnight delivery– Ship with a cold pack (especially if sampling for

bacteria)


Note: It is important that the lab is achieving the primary and secondary purposes - i.e., they are differentiating between mold growth, odd distributions, and normal fallout.

Tape Surface Sampling with Microscopy

Primary purpose: Determine whether mold is or has been growing on the surface and, if so, what typesSecondary purpose: Note a skewing of the normal distribution of spore typesPros:

– Shows exactly what is there– Can be rushed (analyzed immediately)– Can differentiate between mold growth and normal fallout (important)

Cons:– Surface growth does not provide information on airborne spore levels– Cannot differentiate between some spore types– Cannot assess current viability (usually not important)– Areas with minimal growth may be missed if there is high background debris


Tape Surface Sample, Microscopy:Growth, Hyphae, Spores


Tape Surface Sample, Microscopy:Growth, No Hyphae Present


Tape Surface Sample, Microscopy:Growth, No Spores


Tape Surface Sample, Microscopy:Growth, No Hyphae Present


Report of Tape Sample Analysis


Swab Direct Exam With Chaetomium


Report: Swab Direct Microscopy, Quantitative


Note: The most accurate results are obtained by using multiple media and multiple dilutions• Fungi do not have to be viable to cause allergic responses or cause other irritation• Surface growth does not provide information about airborne spore levels• It is important for the lab to use: Multiple media types and multiple dilutions

Culturable Surface Sampling

Pros:• Assesses viability• Differentiates Aspergillus and Penicillium• Permits further speciation if required

Cons:• Surface growth does not provide information on airborne

spore levels• Misses fungi that:

– Are not viable– Do not compete well– Do not germinate on the media provided

• Results may not reflect what was actually growing on the surface

• Cannot be rushed (analyzed immediately)


Pros:• Assesses viability (needed in

certain situations)• Differentiates Aspergillus and

Penicillium• Permits further speciation when

required

Cons:• Misses fungi that

– Are not viable– Do not compete well– Do not germinate on the

media provided

• Cannot be rushed (cannot be analyzed immediately)

NOTE: Fungi do not have to be viable to cause allergic responses or other irritation.

Culturable Air Sampling – Pros & Cons


Report of Culture Plate Air Sample Analysis


Non-Culturable Air Sampling – Pros & Cons

Pros:• Captures all the spores in the air

– Can accurately characterize environments where fungi are present that:

• Are no longer viable

• Do not culture well

• Do not compete

• Can be rushed (analyzed immediately)Note: Fungi do not have to be viable to cause allergic response or other irritation

Cons:• Cannot assess viability (usually not important)• Cannot differentiate between some spore

types• Background debris may limit visibility


Stachybotrys spores

Penicillium/Aspergillus spores

One Half of a 400X Microscope Field


Spore Traps With Different Background Debris


200x 400x 600x 1000x

Spore Traps at Different Magnifications


Report of Spore Trap Sample Analysis








Fungal Data Interpretation

• Unless you’re an M.D., be aware that interpreting data with respect to heath effects can get you into legal difficulties

• Stick to interpreting:– Is there growth in the building?

– Are people being exposed to fungi?

– Is there evidence of building defects?

• Stick to the original proposed hypothesis:– e.g. “Does this data prove that

people are being exposed to Stachybotrys?”


Interpretation

Numerical Guidelines:• No widely accepted standards

exist for any type of fungal sampling (surface, dust, aerosol)

In 1986, 1987, and 1989, the ACGIH published proposed numerical guidelines for interpretation. In 1999, they took them back. It is unlikely that scientifically valid, widely accepted, numerical guidelines will exist in the near future.


General Interpretive Guidelines

Air samples:• It is helpful to compare the inside and outside

air if false positive and negatives are considered.

• The inside sample “should” have:– Generally lower quantities– Same fungal types– Same basic distribution– Look for an order of magnitude difference– Look for “indicator fungal types”– Consider exchange rates of the indoor air with

the outside– Consider activity levels, inside and out– Consider weather– Consider concentrations of the fungi present


Interpretation Sample – Project A


Interpretation Sample – Project B


False Positive Results

• Less well recognized within the industry

• Harder to explain• Important to understand• The aim of the studies

presented here


Effects of Vacuuming


Making the Bed


Moldy Apple Pie Toss


Moldy Scone Toss


Moldy Persimmon Toss


Moldy Food and Total Spore Levels


Interpretation Sample – Project C


In General:

• Data cannot be interpreted in the absence of an understanding of the field conditions and why the data were collected.

• Some system of sanity checking, given the constraints stated earlier, can help guide, train, and educate new investigators.

• An improvement can be made over I/O ratios.


Existing Methods

• Many guidelines for interpretation of indoor/outdoor data have been published – Indoor/Outdoor ratios

• With any ratio > X indicating a problem. Common values for X = 1,5,10

• Requiring a minimum concentration of spores and a higher ratio to indicate a problem.

• Using specific types of spores

• Spearman Rank Correlation • Agreement ratios • Subtracting outdoor concentrations

of specific fungi from those indoors


MoldScoreTM

• We have – Used an aerobiological approach

to develop the MoldScoreTM

– Tested the MoldScoreTM against the opinion of experts

– Reevaluated and updated the algorithms

– Compared MoldScoreTM to • Indoor/outdoor ratios

• Spearman Rank Correlation

• Agreement Ratio


Data Source

• 100 pairs of inside and outside samples

• All included indoor and outdoor spore trap samples

• Distributed over the US from Alaska to Florida

• Distributed throughout the year• Wide range of spore counts and

inside to outside ratios


Agreement Ratio vs. Experts


Spearman’s Rank Correlation vs. Experts


Converted I/O vs. Experts


A2 vs. A1


MoldScoreTM vs. Experts


Data Set 798, June in FL


Ratings for Data Set 798

• Agreement ratio =0.76

• Spearman’s = Yes (similar, OS source)

• Max I/O = 1.9

• MoldScore = 17

• Expert avg. = 11


Data Set 613, July in FL





• Max I/O = 2.9

• MoldScore = 93



Data Set 072, April in OR




• Spearman’s = No (not similar, IS source)

• Max I/O = 15.4

• MoldScore = 100



Data Set 938, December in ND





• Max I/O = 10.9

• MoldScore = 23



Data Set 009, November in FL





• Max I/O = 3.4

• MoldScore = 63



Data Set 561, September in AK





• Max I/O = 0.99

• MoldScore = 100



Data Set 320, December in NY





• Max I/O = 2.5

• MoldScore = 65



Conclusions

• Can be a useful sanity check in some situations.

• Results in a score that is an indication of the probability of indoor fungal growth

• Meant to be used in conjunction with a visual inspection by a qualified home inspector

• Can help large epidemiological studies

• Keep working at it


• Outdoor spore populations vary widely over short periods of time especially during changes in weather.

• Understanding this variability is crucial if indoor to outdoor comparison are to be used for data interpretation.

Variability


Sunny Day Aerosol


The Start of Rain


Rainy Day Aerosol


• With limited outdoor sampling, one needs a database with which to evaluate the representativeness of data

• We have developed the MoldRangeTM, which is a database compiled from over 100,000 outdoor spore trap samples collected across the country and throughout the year.

MoldRangeTM


Research

• The MoldRangeTM data base is also useful for research on the outdoor aerosol– Geographic and seasonal prevalence of certain spore types– Factors influencing prevalence

• Because the data base is continually updated, we can also compare changes in prevalence by local, season, and taxonover time.


By Month, by Taxon: Pen/Asp


Arizona, Cladosporium, chronological

1

10

100

1000

10000

100000

0 50000 100000 150000 200000

sample number

spor

es/m

3

By State, by Taxon: Chronological


Arizona, Pen/Asp, chronological

1

10

100

1000

10000

100000

1000000

40000 45000 50000 55000 60000 65000 70000

Project #

spor

es/m

3

By State, by Taxon: Chronological


By Month by Taxon: Alternaria


Air Sampling Data – MoldRANGE™ Sample Report

• Separates mold types found between “generally able to grow indoors” and “seldom found growing indoors”

• Gives typical outdoor range by month

• Gives typical outdoor range by state


EMLab P&K’s IAQ Pocket Guide

• Sampling and regulatory guidelines for: fungi, asbestos, bacteria, and allergens

• Updated MoldRange™ data from over 260,000 spore trap samples

• Data interpretation guidelines

To order a complimentary Pocket Guide, complete the survey after the webinar. The “Thank you” email (sent an hour after the webinar) will include a link to the survey to order your Pocket Guide.


EMLab P&K Mobile App

Access your lab results: Easier. Faster. Anywhere you are.

• Review your projects and reports as they are made available

• Browse EMLab P&K's fungal library• Contact your Project Manager with the tap of

the screen

More details at http://www.emlab.com/mobile


Interpretation Sample – Project D1






Interpretation Sample – Project E1


Interpretation Sample – Project E2


Interpretation Sample – Project F


Interpretation Sample – Project G1




Discussion Case: Desert Community

Criteria being used:• Inside greater than 10 times

outside• Mold found on surface samples

Data:• Inside spore levels of 400

spores/m3

• Outside spore levels varied from below the detection limit to 200 spores/m3

• Stachybotrys was found on occasional surface samples, but not in the air


Discussion Case: Desert CommunityActions taken:• Used as evidence for amplification• Buildings were evacuated• Contents were deemed

contaminated

Commentary:• Airborne levels were normal• No evidence of airborne exposure

to Stachybotrys is present• Outside levels were low (and often

are in the desert)• Data does not support

contamination of contents


Data Management and Presentation

The presentation of the data:• Can make your job easier

or harder• May affect how people

interpret it• May reflect how it is

perceived in court• Can make communication

with your client easier or harder


Air Sampling Data – Standard Graph

• Makes comparison of data between samples easy

• May understate the importance of certain fungal groups, genera, or species

• May not be valuable if fungal counts for one location are very high


Air Sampling Data – Landscape Graph

• Makes comparison of data between samples easy

• May understate the importance of certain fungal groups, genera, or species

• May not be valuable if fungal counts for one location are very high


• Shows typical outside ranges

• Provides some nice statistical “rules of thumb”

MoldSTAT™




Interpretation Sample – Project H1


Interpretation Sample – Project H2








Dust Samples

• May represent settled dust, or growth in the dust

• Get at least 1 teaspoon of dust, if possible

• Control sampling variables:– Location (stay out of walkways)– Vacuuming methods

• Interpretation is less defined:– Wide range of numbers– Weight of background material affects

calculated results– Consider a control sample

• The most accurate results are obtained if the lab uses multiple media and multiple dilutions


Culturable – Bulk, Swab, Dust

• The culturable sample is diluted 1:10, 1:100, 1:1000, and 1:10000. Then the dilutions are plated 6 times on three different media.


Dust Sampling — Fungal Culture Report


Thank you for your time!

Questions about Mold:[email protected]

All other questions:[email protected]

Appendix

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Texas - Houston6310 Rothway St.Houston, Texas 77040phone: 800.651.4802

California - Glendale1010 N. Central Ave.Suite 420Glendale, CA 91202phone: 866.465.6653

California - Irvine17461 Derian AveSuite 100Irvine, CA 92614Phone: 866.465.6653

California - Sacramento880 Riverside ParkwayWest Sacramento, CA 95605phone: 866.888.6653

Virginia - Fairfax3929 Old Lee HighwayUnit 91CFairfax, Virginia 22030phone: 866.871.1984

Washington - Seattle19515 North Creek Parkway N, Suite 100Bothell, WA 98011phone: 866.888.6653

For the most current list of locations, please visit us at www.emlabpk.comContact individual laboratories for service capabilities and scopes of accreditation.


TestAmerica LocationsFor the most current list of locations, please visit us at www.emlabpk.com

For your convenience, you can drop off samples for EMLab P&K at these locations.CALIFORNIA - Pleasanton1220 Quarry Ln.Pleasanton, CA 94566phone: (925) 484-1919

CALIFORNIA - San Bernardino202 E. Airport RoadSuite 140San Bernardino, CA 92408Phone: (909) 370-4707

CALIFORNIA – W. Sacramento880 Riverside PkwyWest Sacramento, CA 95605phone: (916) 373-5600

CONNECTICUT128 Long Hill Cross Rd.Shelton, CT 06484phone: (203) 929-8140

FLORIDA - Jacksonville8933 Western Way, Suite 1Jacksonville, FL 32256phone: (904) 519-9551

FLORIDA - Orlando8010 Sunport Drive, Suite 116Orlando, FL 32809phone: (407) 851-2560

FLORIDA - Pensacola3355 McLemore Dr.Pensacola, FL 32514phone: (850) 474-1001

FLORIDA - Tallahassee2846 Industrial Plaza Dr.Tallahassee, FL 32301phone: (850) 878-3994

FLORIDA - Tampa6712 Benjamin Rd., Suite 100Tampa, FL 33634phone: (813) 885-7427

GEORGIA - Atlanta6500 McDonough Drive, Suite C-10Norcross, GA 30093phone: (678) 966-9991

GEORGIA - Savannah5102 LaRoche AvenueSavannah, GA 31404phone: (912) 354-7858

HAWAII - Honolulu99-193 Aiea Heights Dr.Suite 121Aiea, HI 96701phone: (808) 486-5227

ILLINOIS - Chicago2417 Bond StreetUniversity Park, IL 60484phone: (708) 534-5200

ILLINOIS - Elmhurst655 W. Grand Ave., Suite 205Elmhurst, IL 60126phone: (630) 758-0262

INDIANA - IndianapolisStutz Business Center212 W. 10th Street, Ste A-205Indianapolis, IN 46202Phone: (317) 264-9686

INDIANA - Valparaiso2400 Cumberland DriveValparaiso, IN 46383phone: (219) 464-2389

IOWA - Cedar Falls704 Enterprise DriveCedar Falls, IA 50613phone: (319) 277-2401

IOWA - Davenport736 Federal St., Suite 2202Davenport, IA 52803phone: (563) 323-7944

LOUISIANA - Baton Rouge6113 Benefit Dr.Baton Rouge, LA 70809phone: (225) 755-8200

MARYLAND - Baltimore5710 Executive Drive, Suite 106Baltimore, MD 21228phone: (410) 869-0085

MASSACHUSETTS - Boston240 Bear Hill Rd., Suite 104Waltham, MA 02451phone: (781) 466-6900

MASSACHUSETTS - Westfield53 Southampton RoadWestfield, MA 01085phone: (413) 572-4000

MICHIGAN - Brighton10448 Citation Drive, Suite 200Brighton, MI 48116Phone: (810) 229-2763

MINNESOTA - Minneapolis7204 West 27th Street, Suite 114St. Louis Park, MN 55426phone: (800) 593-8519

ALABAMA - Mobile900 Lakeside DriveMobile, AL 36693phone: (251) 666-6633

ALASKA - Anchorage2000 W. International Airport Rd., Suite A10Anchorage, AK 99502phone: (907) 563-9200

ARIZONA - Phoenix4625 E. Cotton Center Blvd., Suite 189Phoenix, AZ 85040phone: (602) 437-3340

ARIZONA - Tucson1870 W. Prince Road, Suite 59Tucson, AZ 85705phone: (520) 807-3801

CALIFORNIA – Costa Mesa3585 Cadillac Ave, Suite ACosta Mesa, CA 92626phone: (714) 258-8610


TestAmerica Locations (cont’d)

NEW YORK - Albany25 Kraft Ave.Albany, NY 12205phone: (518) 438-8140

NEW YORK - Buffalo10 Hazelwood Drive, Ste. 106Amherst, NY 14228phone: (716) 691-2600

NEW YORK - New York City47-32 32nd Place, Suite 1141Long Island City, NY 11101Phone: (347) 507-0579

NEW YORK - Syracuse118 Boss Rd.Syracuse, NY 13211phone: (315) 431-0171

NORTH CAROLINA - CharlotteI-85 South Bldg.2858 Queen City Dr., Suite BCharlotte, NC 28208phone: (704) 392-1164

NORTH CAROLINA - Raleigh101-F Woodwinds Industrial CourtCary, NC 27511phone: (919) 380-9919

For the most current list of locations, please visit us at www.emlabpk.comFor your convenience, you can drop off samples for EMLab P&K at these locations.

OHIO - Cincinnati11416 Reading RoadCincinnati, OH 45241phone: (513) 733-5700

OHIO - Dayton4738 Gateway CircleDayton, OH 45440Phone: (937) 294-6856

OHIO - North Canton4101 Shuffel Street NWNorth Canton, OH 44720phone: (330) 497-9396

OREGON - Portland9405 SW Nimbus AvenueBeaverton, OR 97008phone: (503) 906-9200

PENNSYLVANIA - King of Prussia1008 W. Ninth Ave.King of Prussia, PA 19406phone: (610) 337-9992

PENNSYLVANIA - Pittsburgh301 Alpha DrivePittsburgh, PA 15238phone: (412) 963-7058

SOUTH CAROLINA - Charleston1436-A North Point LaneMt. Pleasant, SC 29464phone: (843) 849-6550

TENNESSEE - Knoxville5815 Middlebrook PikeKnoxville, TN 37921phone: (865) 291-3000

TENNESSEE - Nashville2960 Foster Creighton Dr.Nashville, TN 37204phone: (615) 726-0177

TEXAS - Austin14050 Summit Dr., Ste. A100Austin, TX 78728phone: (512) 244-0855

TEXAS - Beaumont4400 Lawndale Ave.Groves, TX 77619phone: (409) 540-5302

TEXAS - Corpus Christi1733 N. Padre Island DriveCorpus Christi, TX 78408phone: (361) 289-2673

MISSOURI - Eureka1699 West Fifth Street, #200Eureka, MO 63025Phone: (314) 302-8354

MISSOURI - Kansas City601 NW 39th StreetBlue Springs, MO 64015phone: (800) 276-1286

MISSOURI - St. Louis13715 Rider Trail NorthEarth City, MO 63045phone: (314) 298-8566

NEW JERSEY - Edison777 New Durham RoadEdison, NJ 08817phone: (732) 549-3900

NEW JERSEY - South Jersey520 Fellowship Rd., Suite A-106Mt. Laurel, NJ 08054phone: (856) 222-1990

TEXAS - San Antonio404 E. Ramsey, Suite 208San Antonio, TX 78216phone: (210) 344-9751

VERMONT - Burlington30 Community Drive, Suite 11South Burlington, VT 05403phone: (802) 660-1990

VIRGINIA - Virginia Beach5135 Cleveland StreetVirginia Beach, VA 23462phone: (757) 671-1291

WASHINGTON - Richland2800 George Washington WayRichland, WA 99354phone: (509) 375-3131

WASHINGTON - Spokane11922 E. 1st Ave.Spokane, WA 99206phone: (509) 924-9200

WASHINGTON - Tacoma5755 8th Street EastTacoma, WA 98424phone: (253) 922-2310


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Accreditation: A formalized recognition that a facility meets the AIHA, or other accrediting authorities, policy requirements to carry out defined tasks or specific types of tests. See also Certification.

Accredited Laboratory: A testing laboratory that has been evaluated and granted accreditation covering a specified measurement or task, usually for a specific property or analyze, and for a specified period of time.

Accuracy: The degree of agreement between an observed value and an accepted reference value or statistically defined criteria. Accuracy includes a combination of precision and bias. See Precision and Bias.

Agar: Musilagenous substance (galactan) extracted from some algae used to grow fungi and bacteria.

Aliquot: See Subsample.

Analysis: A qualitative or quantitative determination of a substance or material.

Analytical Method: A standardized set of procedures required to prepare a sample for analysis and analyze a sample to obtain a qualitative or quantitative determination. See also Standard Operating Procedure (SOP).

Approved Signatory: A person who is recognized by the laboratory as competent to adopt SOP’s, approve change control documents and other internal QC documents, verify the integrity of results, and introduce policies related to Quality Assurance.

Audit Trail: A process of documentation that allows for the tracking of a sample through the laboratory, including the time, date, and identification of the individual(s) responsible for performing an action during the receiving, log-in, sample preparation, and analysis of a sample. See also Sample Tracking.

Autoclave: A device that sterilizes using pressurized steam for a prescribed time period. Generally autoclaves run at 121º C and 15 PSI. Autoclaving allows for sterilization at lower temperatures than dry heat.

Bacterium (pl. bacteria): Heterogenous group of unicellular prokaryotic organisms.

Bacteriology: The scientific study of bacteria.

Bias: A systematic error manifested as a consistent positive or negative deviation from the known true value.

EMLab P&K General Glossary


Bioaerosol: Airborne particles that are either alive or originate from living organisms. Bioaerosols include microorganisms (i.e. culturable, non-culturable, and dead microorganisms) and fragments, toxins, and particulate waste products from all varieties of living things.

Biohazard: An object that contains infectious or dangerous organisms, or any object that contains an inordinate amount of organisms.

Blank: An unexposed sampling media or reagent(s), not taken to the field or shipped, but carried through the complete sample preparation and analytical procedure. The blank is used to assess possible background contamination from the analytical process.

Blind Sample: A sample submitted for analysis with a composition and identity known to the submitter, but unknown to the analyst, and used toevaluate proficiency in the execution of the measurement process.

Bulk Sample: Any non-liquid, solid matrix submitted for analysis that cannot be categorized as a soil, dust, or spore trap. Some examples of bulks include carpet, drywall, carpet backing, particleboard, wood, etc. Ideally bulks samples should be reported as a surface area instead of a unit of mass.

Calibration: To determine, by measurement or comparison with a standard, the correct setting on a meter or other instrument. See Calibration curve.

Calibration Curve: A graphical relationship between the known values for a series of calibration standards and instrument responses. The levels ofthe calibration standards should bracket the range of measurements.

Certification: Procedure by which a third party gives written assurance that the process of testing and evaluating against specifications designed to document, verify and recognize the competence of a person, organization, or other entity to perform a function or service conforms to specified requirements. See Accreditation.

Certified Reference Material (CRM): A reference material that has one or more of its property values established by a technically valid procedure, and is accompanied by or traceable to a certificate or other documentation issued by a certifying body. See Reference material.

Chain of Custody: An unbroken trail of records that documents the location of samples, associated data, and records. See also Audit Trail.

Change Control (CC): The process by which changes are made to any controlled document that defines procedures, processes, products, etc. The process involves formally updating all associated documents and officially removing and archiving all the non-updated versions of the document.

Client: Any person or organization that engages the services of the laboratory.

EMLab P&K General Glossary (continued)


Communications: Transmission of information by any means including verbal, mail, and electronic.

Computer Records: Files containing data that can be retrieved when needed and documentation of alterations to data. Computer records should not allow unauthorized access to data or the undocumented amendment of data.

Conclusion: A determination based on the analysis and interpretation of data collected from the testing of a hypothesis. See also Hypothesis Testing.

Contamination: The introduction of non-indigenous organisms or substances into an environment, or the introduction of organisms or substances into a previously sterile environment.

Control Chart: A graph of some measurement plotted over time or sequence of sampling, together with control limit(s) and, usually, a central line and warning limit(s).

Corrective Action: A process that addresses a Work Out of Specification, Client Contact, or Continuous Improvement event that is rated as significant by the management team and requires a change to the way a process or the work is performed, or an adaptation to the Quality Control System associated with the step or process.

Culturable Air Analysis: A technique that assesses the amount of viable organisms present in the air. The technique allows for the differentiation of Penicillium and Aspergillus species, and permits further speciation when required. The technique misses fungi that are not viable, do not compete well, or do not germinate on the selected medium.

Direct Examination (Surface) Analysis: A technique that assesses whether mold is growing on a surface, and if there is mold growth, what organisms are present. A direct exam also notes if there is skewing of the distribution of mold types. The technique shows exactly what is found on the surface and can differentiate between growth and normal fallout. The technique cannot differentiate between certain spore types, does not assess the airborne levels of spores, or give any information about the viability of organisms.

Document Control: A system of protecting the integrity of processes and systems in the laboratory by uniquely identifying and monitoring the alteration to documents within the laboratory. New documents and revisions or changes to documents are signed for authorization by a designated signatory before implementation in the laboratory. See also Approved Signatory.

Duplicate Analyses: The analyses or measurements of the variable of interest performed identically on two sub-samples of the same sample. The results from duplicate analyses are used to evaluate analytical or measurement precision but not the precision of sampling, preservation or storage internal to the laboratory.



Duplicate Samples: Two samples taken from and representative of the same population and carried through all steps of the sampling and analytical procedures in an identical manner. Duplicate samples are used to assess variance of the total method including sampling and analysis.

Dust Sample: A sample matrix generally collected using a vacuum device and a cassette, or other collection device, containing a filter or filtering matrix. Typically dust samples are reported per unit of area vacuumed or per unit mass.

Endotoxins: Toxins that are confined inside the microorganisms and released only when microorganisms are broken down or die.

Environmental Microbiology: The area of microbiology that focuses on the biology, physiology, ecology and sampling and analysis of microorganisms inhabiting or affecting air, water, soil and other natural or man-made substances and/or systems in a variety of work environments, and that may contribute to adverse health effects.

EMLab P&K Accreditation Program (EMLAP): This AIHA program is intended for the accreditation of environmental microbiology laboratories.

Environmental Microbiology Proficiency Analytical Testing (EMPAT): Required proficiency testing program in the core business of an EMLabP&K .

Equipment: All physical items (including software and instruments) in a facility used in the process of analytical testing.

Equipment Log: A chronological record of preventive and emergency maintenance performed on an analytical instrument. The logs include a record of calls, service technician summaries, records of calibration by the manufacturer, and other information as required by these policies.

Facility: A permanent, temporary or mobile operation established for the purpose of performing laboratory testing.

Field Blank: An analyte-free matrix (i.e. spore trap cassette, media plate, tape, etc.) carried to the sampling site, kept sealed or un-exposed throughout the sampling process, returned to the laboratory, treated as a sample, and carried through all steps of the analysis.

Fungus (pl. fungi): Eukaryotic organisms without plastids, nutrition absorptive, never phagotrophic, lacking an amoeboid pseudopodial phase.

Hardcopy: Information or data recorded on a paper form or other physical document. See also Computer Records and Single Strike Out

Hypothesis Testing: The process of developing a tentative explanation that accounts for a set or facts that is either proved or disproved by subsequent investigation.



Incubation period: The time between inoculation and the development of visible symptoms

Industrial Hygiene Experience: A person’s history of onsite evaluation of a variety of occupational environments to include inspection, monitoring, laboratory analysis and control recommendations regarding actual or potential occupational exposure to conditions likely to result in adverse health effects.

Instrument: A device used for observation or measurement that yields test results.

Intra-analyst Comparisons: A method for measuring and improving analyst precision, and to a certain extent accuracy, by having multiple analysts analyze one sample. Raw counts are statistically analyzed to determine the degree of variance in counting amongst analysts.

Internal Quality Control: Routine activities and checks, such as periodic calibrations, duplicate analyses, and intra-analyst comparisons, that are performed during routine internal procedures to control the accuracy and precision of measurements.

Laboratory: A place within a permanent, mobile or temporary facility used for the purpose of analysis.

Laboratory Assessment: An onsite evaluation of a laboratory for the purpose of conducting a technical systems audit to assess compliance with AIHA accreditation requirements.

Laboratory Accreditation Committee (LAC): A group of qualified individuals responsible for carrying out the technical business of an AIHA Laboratory Accreditation Program (LAP). Each individual LAP will have an LAC.

Laboratory Quality Assurance Committee (LQAC): General term referring to one or more committee(s) associated with any analytical program.

Laboratory Quality Assurance Program (LQAP): General term referring to any AIHA program or programs established to maintain the highest possible standards of performance for analysts and/or laboratories analyzing samples and evaluating exposures to hazardous agents.

Lot: A set of samples submitted together for laboratory analysis that can be treated as one or more batches. Lot may also refer to a batch of sampling media or reagents manufactured at the same time.



Macrofungi: See also Mushroom.

Matrix: The component or substrate (e.g., spore trap, air or bulk) that contains the analyte of interest.

Method Detection Limit (MDL): The minimum concentration of an analyte that, in a given matrix and with a specific method, has a 99 percent probability of being identified, qualitatively or quantitatively measured, and reported to be greater than zero.

Method Performance: A general term used to document the characteristics of a method. These characteristics usually include method detection limits, linearity, precision, accuracy and bias and uncertainty of measurement. See Acceptance limits.

Microfungus: A fungus whose morphological features are visible by high magnification compound microscope. e.g. Cladosporium, Aspergillus etc.

Mold: A microfungus having a well-marked mycelium or spore mass; a subset of fungi.

Mushroom: A fleshy basidioma, usually stalked and with a cap (pileus) beneath which gills or fleshy tubes are covered or lined with the hymenium.

Mycology: The scientific study of fungi.

Mycotoxins: Secondary metabolites (toxin) produced by fungi.

Policy: An organization’s written statement of commitment to implement a management program element.

Precision: The degree to which a set of observations or measurements of the same property, usually obtained under similar conditions, conform to themselves. Precision is often expressed as standard deviation, variance or range, in either absolute or relative terms.

Preventive Action: A planned activity to identify, recognize and control potential sources of nonconformance.

Procedure: A written set of instructions that describes how to implement a policy requirement, or how to carry out a specific task

Proficiency Testing: Refers to any proficiency analytical testing program(s), such as the programs established under the



Quality: A characteristic of a high standard of service including reliability and traceability.

Quality Assurance: An integrated system of activities involving planning, quality control, quality assessment, reporting, and quality improvement to ensure a product or service meets defined standards of quality within a stated level of confidence.

Quality Assurance Coordinator: See Quality manager.

Quality Control (QC): Technical activities whose purpose is to measure and control the quality of a product or service so that it meets the needs of users. The aim is to provide quality that is satisfactory, adequate, dependable and economical.

Quality Manager: The manager of the quality system. The quality manager is independent of the analyst (for a specific sample set) and reports directly to the highest level of management.

Quality Manual: A document stating the quality policy, quality system and internal quality control procedures of the laboratory.

Quality System: See Quality assurance.

Record Control: The procedures used to identify, collect, index, access, file, store, maintain, and dispose of quality and technical documents.

Reporting Limits: The maximum or minimum levels, or quantities of a target analyze, that can be quantified with the certainty required by the data user.

Reference Material: A material or substance, one or more properties of which are sufficiently homogeneous and well established to be used to monitor instrument and method performance. AIHA PAT samples may be used as reference materials.

Relevant Degree: A program of collegiate study that is appropriate to the applicable accreditation program.

Reproducibility: The extent to which a method, test or experiment yields the same or similar results when performed on sub-samples of the same sample by different analysts or laboratories.

Revision Number: A system of tracking new editions to a document or form. See also Document Control.

Root Cause Analysis (RCA): A process used to identify the cause of a problem or error. The process involves identifying all of the portions of a problem and through the process of elimination, narrowing the field down until a cause can be agreed upon by consensus.



Sample Log: A document where sample identification, date received, client, etc., are noted when samples arrive at the laboratory. The log is part of the sample tracking system. See Sample tracking.

Sample Tracking: A document system of following a sample from receipt at the laboratory, through sample processing and analysis, to final reporting. The system includes unique numbering, or bar coding labels, and the use of a Sample Log.

Sporulation: Process by which spores are produced.

Standard Operating Procedure (SOP): A written document that details the procedures of an operation; an analysis or action whose techniques and procedures are thoroughly prescribed, and which are accepted as the procedure for performing certain routine or repetitive tasks.

Standard Reference Material: A certified reference material produced by the U.S. National Institute of Standards and Technology (NIST) and characterized for absolute content independent of analytical method. It is accompanied by a certificate that reports the results of the characterization and describes the intended use of the material.

Standardization: The process of establishing the quantitative relationship between a known mass of target material (for example, the measurementsystem or instrument response). See Calibration and Calibration curve. The term may also refer to activities that establish provisions for common and repeated use of accreditation policies to achieve an optimum level of conformity.

Stock Solution: A concentrated solution of analyte(s) or reagent(s) prepared and verified by prescribed procedure(s), and used for preparing calibration standards. See Calibration standard.

Subsample: A representative portion of a sample; a subsample may be taken from any location or a field sample; in analytical chemistry, an“aliquot.”

Suspension: A temporary removal of the accredited status of a laboratory when it is found to be out of compliance with specific program requirements.



Taxonomy: The science of classification.

Technical Systems Audit: A thorough, systematic, onsite, qualitative evaluation of facilities, equipment, personnel, training, procedures, record keeping, data validation, data management and reporting aspects of a total quality system.

Test: A technical operation that consists of determining one or more elements in a sample according to a specified procedure.

Test Method: Specified technical procedure for performing a test.

Traceability: The process of documenting the value of a reference material or standard as related to NIST standards or equivalent through an unbroken chain of comparisons with stated uncertainties.

Training: A series of documented tasks related to learning

Uncertainty of Measurement: Result of the evaluation aimed at characterizing the range within which the true value of a test result is estimated to lie, generally within a given likelihood.

Validation: The process of confirming specified method performance criteria.

Verification: Confirmation by examination and provision of evidence that specified requirements have been met.

Work Out of Specification (WOOS): Work or processes that fall outside of the specifications of the internal Quality Control or deemed by the personnel performing the task as not meeting the requirements of normal operation.


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