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Allergens and Bacteria
Webinar
Will begin at 9:00 a.m. (PST)
Participants will be in listen only mode.
Download the PDF of this presentation (case sensitive):
http://www.emlab.com/m/media/CET-webinar-day2.pdf
Allergens
Bacteria
Risk Communication
Harriet Burge, Ph.D.
Copyright © 2007-2013 EMLab P&K. All rights reserved. 2
Continuing Education Units (CEUs)
To receive a certificate of attendance, you must complete the survey after the webinar:
• Click on the survey link in the “Thank you” email (sent 1 hour after this webinar).
• Complete survey by this Friday, March 15, 2013.
• You will receive an email in 2-3 weeks with instructions when your certificate is ready.
Copyright © 2007-2013 EMLab P&K. All rights reserved. 3
Seminar Agenda – Day 2
1. Allergens
2. Bacteria a. Overview of bacteria
b. Summary of some adverse health effects due to bacteria a. Legionella
b. Endotoxin
c. MRSA
d. Sewage
3. Sampling and analysis
4. Risk communication
Allergens
Participants will be in listen only mode.
Otherwise, the background noise makes
effective communication impossible.
Harriet Burge, Ph.D.
Copyright © 2007-2013 EMLab P&K. All rights reserved. 5
The Immune System
• Passive immunity Physical barriers (skin)
Low pH of our stomach
Coughing
• Active immunity
• Mediators Specific cell types
Antibodies
• Types Antigen exposure
Transfer from mother
Acquired
Copyright © 2007-2013 EMLab P&K. All rights reserved. 6
A Few Terms
• Antigen: A substance that stimulates an immune response, esp. production of antibodies.
• Antibody: One of a variety of proteins called immunoglobulins
– normally present in the body or produced in response to an antigen
– IgA, IgD, IgE, IgG, IgM
Source: http://en.wikipedia.org/wiki/Antigen
Copyright © 2007-2013 EMLab P&K. All rights reserved. 7
Internal Risk Factors For Infection
Poorly Developed or Impaired Immunity
• Age. Neonates and the elderly are at increased risk of
bacterial infections.
• Nutritional status. Malnutrition results in a depressed
immune system
• Genetic predisposition. The Human Genome Project
increased our ability to locate specific genes related to
infectious disease susceptibility ( Bentley, DR, 2000).
• Immunosuppression via disease or medications
• Lack of induced specific antibodies
Copyright © 2007-2013 EMLab P&K. All rights reserved. 8
Hypersensitivity Diseases
• Disease states caused by antigen exposure and subsequent sensitization
– Allergy
• Immunoglobulin E (IgE)
– Hypersensitivity pneumonitis
• Immunoglobulin G (IgG)
Copyright © 2007-2013 EMLab P&K. All rights reserved. 9
Allergy
• Allergy is an inappropriate over-reaction of our immune system.
• IgE antibodies are produced by B cells in response to allergen exposure
• IgE molecules attach to Mast cells.
• Mast cells release Histamine and other chemicals
Copyright © 2007-2013 EMLab P&K. All rights reserved. 10
Allergy
• Genetically controlled
• Sensitization thought to require repeated low-level exposures to allergens
• Allergens are complex proteins produced by many different organisms Dust mites, cockroaches
Cats, dogs, other pets
Rats, mice
Fungi
Plants
• Symptoms include upper respiratory (hay fever), lower respiratory (asthma), systemic (anaphylaxis), and skin (hives).
Copyright © 2007-2013 EMLab P&K. All rights reserved. 11
IgG
• IgG generally helps with antibody based protection against pathogens.
Allergens may also lead to high levels of IgG.
Sensitization is thought to require intense exposure to the allergen and may occur relatively quickly.
Sensitization may lead to flu-like symptoms.
Continued exposure can lead to hypersensitivity pneumonitis
Copyright © 2007-2013 EMLab P&K. All rights reserved. 12
Some Indoor Sources of Allergens
Organism Some allergens Some sources
within organism
Some exposure
sources
Dust mites Der f1, Der p1, Blo
t1, etc. Fecal material
Dust, especially
bedding
Cockroaches Bla g1, Bla a1 Skin secretions Dust, especially,
kitchens
Cats Fel d1, etc. Skin secretions Airborne,
accumulates in dust
Dogs Can f1 Skin secretions Airborne,
accumulates in dust
Rodents Rat r1, Mus m1 Urine Airborne,
accumulates in dust
Fungi Alt a1, Pen c1,
Asp v1, etc.
Digestive
enzymes
Airborne with spores,
accumulates in dust
Copyright © 2007-2013 EMLab P&K. All rights reserved. 13
Dust Mites
• Two most common house dust mites:
– American dust mite (Dermatophagoides
farinae)
– European dust mite (Dermatophagoides
pteronyssinus)
• Very small, invisible to the unaided eye.
• Live for approximately 3-4 months.
• Prefer warm, moist areas (mattresses,
pillows, carpeting and upholstery)
• Cause of asthma and allergy:
– Approximately 85% of asthmatics are allergic
to dust mite allergens
– Evidence exists for a causal relationship
between dust mite exposure and the
development of asthma
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Cockroach
• Two most common indoor species of
cockroach in North America are:
– German cockroach (Blatella germanica)
– American cockroach (Periplaneta americana)
• Widely distributed in homes and schools
• Found throughout homes, including beds,
furniture, and carpets - highest levels are
typically found in kitchen
• Exposure is more likely in the bedroom
because the allergens are aerosolized when
getting into bed
• ~20% of homes with no evidence of
cockroach infestation have significant levels
in settled dust
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Cat
• The household cat species is Felis
domestica
• Cats are pets in 27% of U.S. households
• Cat allergens are very sticky and can
remain airborne for long periods of time
• High levels are often found on walls and
other surfaces within homes
• Cat allergens are easily carried from
home to home, office, school, etc.
• The vast majority of homes contain cat
allergen, even if a cat has not lived there
• Cat allergen is produced in cat
sebaceous, salivary and anal glands
Copyright © 2007-2013 EMLab P&K. All rights reserved. 16
Dog
• The household dog species is Canis
familiaris
• Dogs are pets in 31% of U.S. households
• Dog allergens are associated with particles
smaller than 5µm, which are easily
aerosolized and can remain airborne for long
periods of time
• Carpeting, bedding, and upholstered
furniture are typical reservoirs for deposited
dog allergen
• Dog allergen is an important cause of
asthma
• There are over 100 million dogs and cats in
the U.S. and more than 15 million Americans
have allergic reactions to dog allergen
Copyright © 2007-2013 EMLab P&K. All rights reserved. 17
Rodent (Mouse and Rat)
• Two known sources of rodent allergen are:
– Rat (Rattus norvegicus)
– Mouse (Mus musculus)
• High concentrations of these allergens are
present in the rodent’s urine.
• May be an important cause of asthma and
allergic rhinitis.
• Exposure to rodent allergens have been
linked to increased asthma symptoms
among research animal lab workers.
• A major study on asthma among inner-city
children found that nearly 20% of
asthmatic children had been sensitized to
rats and 15% were sensitized to mice.
Copyright © 2007-2013 EMLab P&K. All rights reserved. 18
Mold
• Most molds produce potential
allergens
• Mold allergens are proteins. Two
commonly identified mold allergens
are Alt a1 and Asp f1
• Well documented evidence exists for
mold as a cause of allergic disease
(asthma, hay fever, upper respiratory
allergies, hypersensitivity pneumonitis)
• 30 to 35% of the population can have
allergic response to the environment.
1/3 of these (10% of the population)
may have fungal allergies
Copyright © 2007-2013 EMLab P&K. All rights reserved. 19
Sampling for Allergens
• Sample using a dust
cassette.
• Approximately 150mg or 1
teaspoon of dust is required
for the analysis
• Dust mite allergen analysis is
time sensitive and should be
sent using a cold pack or a
cooler to prevent the
increase of allergens in
transit
Copyright © 2007-2013 EMLab P&K. All rights reserved. 20
How Are Allergen Samples Analyzed?
• Allergens are analyzed by
ELISA (Enzyme-Linked
ImmunoSorbent Assay)
• ELISA is a highly sensitive
and specific test that uses
monoclonal antibodies that
latch on to the specific
allergens
• Results are expressed as
micrograms (µg) of allergen
per gram (g) of dust
Copyright © 2007-2013 EMLab P&K. All rights reserved. 21
How Do I Interpret the Results?
Critical levels for sensitization
Dust mite 2.0µg/g of dust
Cockroach (Bla g1) 0.2 units/g of dust
Cat 1-8µg/g of dust
Dog Sufficient evidence is
Mouse not yet available for
Rat establishing risk levels
for these allergens.
Critical levels for exacerbation
Dust mite 10.0µg/g of dust
Cat >8µg/g of dust
Cockroach (Bla g1) 1 to 2 units/g of dust
Copyright © 2007-2013 EMLab P&K. All rights reserved. 22
Dust Mite Control
• Keep humidity below 50%
• Vacuum regularly and discard bags immediately; Replace carpet with hard flooring if possible
• Wash bedding every week in hot water (54°C); Use allergen free mattress and pillow covers
• Minimize upholstered furniture and curtains or drapes
• Minimize dust and dust catchers
Copyright © 2007-2013 EMLab P&K. All rights reserved. 23
Pest control
• Keep kitchens clean and dry
• Use cockroach traps; if you see cockroaches, call an
exterminator
• Seal up openings for mouse/rat entry
• If you see mice or rats, call an exterminator
Copyright © 2007-2013 EMLab P&K. All rights reserved. 25
EMLab P&K’s IAQ Pocket Guide
• Sampling and regulatory guidelines for: fungi, asbestos, bacteria, and allergens
• Updated MoldRange™ data from over 350,000 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.
Copyright © 2007-2013 EMLab P&K. All rights reserved. 26
Outline for Bacteria
• Bacteria in the natural environment
• Nature and classification
• Overview of sampling and analytical methods for bacteria
• Health effects
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Distribution and Role of the Bacteria in
the Natural Environment
• Ubiquity:
– “We live now in the ‘Age of Bacteria.’ Our planet has
always been in the Age of Bacteria, ever since the first
fossils — bacteria, of course — were entombed in rocks
more than 3 billion years ago. Bacteria are — and always
have been — the dominant forms of life on Earth”
— Steven J Gould
– All surfaces on earth and all water contain bacteria
– Approximately five nonillion (5x1030) bacteria on Earth,
forming much of the world's biomass.
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• A teaspoon of soil may contain
100 million to 1 billion bacteria.
• Decomposers: can break down
pesticides and pollutants in soil
• Mutualists form partnerships
with plants. The most well-
known of these are the
nitrogen-fixing bacteria (ex.
Rhizobium)
• Plant pathogens: Xymomonas
and Erwinia species, and
species of Agrobacterium that
cause gall formation in plants
Soil
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• A million (106) bacterial cells in a milliliter of fresh water
• 5x108 in an 8 ounce glass of water
– Pseudomonas, Flavobacterium and Acinetobacter
Water
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Air
• Over 2,000 different kinds
of bacteria may be present
in the air on any given day
(Gary Anderson).
• Bacteria are attracted to
bubbles in water; Bubbles
bursting at surface release
bacteria into the air.
• Bacteria may reproduce
within airborne droplets • Bacteria, globally
distributed in the
atmosphere, play a
large role in formation
of snow and rain.
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Plant Surfaces
• In the air, water clumps
around bacteria forming
condensation nuclei,
leading to rainfall.
• Precipitation returns
bacteria to the ground and
onto leaf surfaces where
they multiply.
• Epiphytic bacteria can
increase water permeability
of leaf cuticles, increasing
the availability of water and
dissolved compounds.
• Bacteria multiply on the
surface of plants and are
aerosolized by wind and
rain action.
Copyright © 2007-2013 EMLab P&K. All rights reserved. 32
• There are at least ten times
as many bacteria as human
cells in the body
• In a 200-pound adult, these
organisms can weigh a
combined 2 to 6 pounds.
http://www.scientificamerican.com/arti
cle.cfm?id=microbiome-graphic-
explore-human-microbiome
• This interactive website
shows the kinds of bacteria
on/in different human parts.
The Mammalian Body
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Biofilms: A Larger Structure
• In nature, most bacteria
are bound to surfaces in
biofilms
• Few microns to half a
meter (yes, meter) in
depth.
• Complex arrangement of
cells & extracellular
components including
networks of channels to
diffuse nutrients.
Source: wikipedia.org/wiki/Biofilm
• Shower biofilm
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• Naming bacteria
• Morphology
– Macroscopic
– Microscopic
• Classification
• Physiology
Introduction To Bacteria
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Bacterial Diversity
Source: http://en.wikipedia.org/wiki/Bacteria#Multicellularity
Copyright © 2007-2013 EMLab P&K. All rights reserved. 36
Naming Bacteria
Binomials:
– Genus: Escherichia
– Species: coli
– Genus: Staphylococcus
– Species: aureus
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• DNA: loosely organized, no
membrane bound nucleus
• RNA: in cytoplasm
• Ribosomes: make protein
from amino acids under
instruction from RNA
• Plasma membrane:
lipid/protein layer with
selective permeability Source: http://en.wikipedia.org/wiki/File:Average_prokaryote_cell-_en.svg
• Cell wall:
– Peptidoglycan
– Lipopolysaccharide (Gram negative)
• Capsule (polysaccharides)
Bacterial Cell Structure
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Size
– 0.1 to about 600 µm over a
single dimension
– Escherichia coli: 1.1 to 1.5
µm by 2.0 to 6.0 µm
– On surfaces, usually present
as colonies with a few or
millions of cells
– When airborne, usually on
rafts (e.g., skin scales) (>10
µm) or in droplet nuclei (one
or more bacteria surrounded
by dried mucous) (>2 µm).
Source: http://en.wikipedia.org/wiki/File:Relative_scale.svg
Morphology
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Shape
– Cocci: Spherical
• Staphylococcus aureus
– Bacilli: Rod shaped
• Escherichia coli
• Pseudomonas
aeruginosa
– Spirilli: Spiral rods;
• Borrelia burgdorfii
• Helicobacter pylori
– Filamentous: long
branching strands
• Thermoactinomyces
vulgaris
Source: http://en.wikipedia.org/wiki/File:Bacterial_morphology_diagram.svg
Morphology
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Practical Classification
Bacillus
Corynebacterium
Staphylococcus
Streptococcus
Streptomyces
Micropolyspora
E. Coli
Pseudomonas Neisseria Vibrio
Rods Cocci
Gram positive
Common
bacteria
Filaments Rods Cocci
Gram negative
Spirals
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The Gram Stain
• Gram positive
– Cells with a peptidoglycan
outer wall
• Staphylococcus
• Bacillus
• Gram negative
– Cells with a
lipopolysaccharide outer
wall
• Pseudomonas
• “coliforms”
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Acid Fast
• Cells with mycolic acid in
outer wall: retain a dye
when treated with acid.
All acid fast bacteria are
Gram positive.
– Mycobacterium
– Nocardia
– Some amoebal cysts
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Physiology
• Professor Alan Parsons
and Dr. Richard Heal of
QinetiQ Ltd, claim to have
shown that physically
separated colonies of
bacteria can transmit
signals conferring
resistance to commonly
used antibiotics
• (volatile organic
compounds)?
Copyright © 2007-2013 EMLab P&K. All rights reserved. 44
• Psychrophiles: Grow
best at cold
temperatures. <10°C
• Mesophiles: Grow best
at medium temperatures.
All infectious bacteria are
mesophiles. 10-40°C
• Thermophiles: Grow
best at hot temperatures.
40-80°+C
Temperature Requirements
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Oxygen Requirements
• Obligate Aerobes: strictly oxidative respiration and
require oxygen for survival.
• Facultative Anaerobes: fermentation in the absence of
O2, or respiration in its presence.
• Obligate Anaerobes: anaerobic fermentation; cannot
survive in the presence of oxygen.
Copyright © 2007-2013 EMLab P&K. All rights reserved. 46
• (volatile organic compounds)?
Volatile Organic Compounds (VOCs)
Copyright © 2007-2013 EMLab P&K. All rights reserved. 47
Sampling and Analysis
• Type of sample collection and choice of analytical
methods depends on:
– Hypothesis or goals (monitoring)
– Expected concentrations
– Agent(s) of concern
– Standards/guidelines
Copyright © 2007-2013 EMLab P&K. All rights reserved. 48
Types of Samples and Relevant
Hypotheses or Goals
• Water
– This water contains sewage organisms
– This water contains Legionella pneumophila
• Surface
– Potential pathogens are falling into wounds during surgery
– Residual contamination is present on these surfaces
• Bulk
– This slime contains Legionella pneumophila
– This humidifier water contains Thermophilic actinomycetes
• Air
– Exposure is occurring to this agent
– This activity produces aerosols containing this agent.
Copyright © 2007-2013 EMLab P&K. All rights reserved. 49
Analytical Methods: Bacteria
• Culture
• Microscopy
• Stains
• DNA methods
• Bioassays
• Immunoassays
• HPLC
• GCMS
Copyright © 2007-2013 EMLab P&K. All rights reserved. 50
Culture
• Requires organism to be alive; samples must be protected from extreme conditions, and from conditions that allow growth during transport.
• Recovers only organisms that can reproduce under the provided conditions
– Best to use broad spectrum medium such as R2A that allows damaged bacteria to recover
• Always underestimates concentrations and diversity
• Hypotheses:
– Potential pathogens are falling into wounds during surgery
– Legionella pneumophila is growing in this humidifier.
Copyright © 2007-2013 EMLab P&K. All rights reserved. 51
Microscopy
• Vital Staining
– Use of stain that differentiates living from dead bacteria
– All cells can be counted and % viable calculated
– Samples must be maintained to minimize growth and death during transport.
• Fluorescence staining
– Allows microscopic or flow cytometric counting of cells; fluorescent vital staining available
• Fluorescent antibody staining
– Allows counting of specific organisms
• Hypotheses:
– The bacterial aerosol has xx living and xx non-living organisms
– This treatment kills bacteria
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Other Kinds of Staining
• Gram stain and Fluorescent gram stains
– This particular population consists of Gram negative bacteria (or not)
• Acid fast staining
– Mycobacterium cells are present in this tissue sample
• Acridine orange (a fluorescent stain)
– Total concentration of bacterial cells in this sample is xx
Mycobacterium tuberculosis (stained red) in tissue (blue).
Source: en.wikipedia.org/wiki/acid-fast
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DNA Methods
• For identification
• For monitoring populations
– This specific strain of Legionella pneumophila is present in
this cooling tower
– Track composition of bacterial populations are present in
this aerosol
– The bacterial population in this biofilm is the same as or
different than the one in a different biofilm.
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Bioassays
• Limulus assay for endotoxin
– Depends on the horseshoe crab
– Quantitative only within each batch of lysate
– Internal controls essential for every assay
• This sample contains more endotoxin than the outdoor air
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Immunoassays for Specific Bacteria
• There are immunoassays for surrogates of bacteria
considered possible biowarfare agents (e.g., Bacillus
globigii as a surrogate for Bacillus anthracis).
• Obviously it is possible since bacteria can stimulate an
antibody (immune system) response. Such methods
have not been widely used.
Copyright © 2007-2013 EMLab P&K. All rights reserved. 56
HPLC, GCMS
• Chemical methods used for measurement of bacterial
and other biological chemicals.
• HPLC: high pressure liquid chromatography
• GCMS: Gas chromatography Mass spectroscopy
• Fatty acid analysis used for identification of bacteria in
bulk samples
Copyright © 2007-2013 EMLab P&K. All rights reserved. 57
To receive a certificate of attendance, you must complete the survey after the webinar:
• Click on the survey link in the “Thank you” email (sent 1 hour after this webinar).
• Complete survey by this Friday, March 15, 2013.
• You will receive an email in 2-3 weeks with instructions when your certificate is ready.
Continuing Education Units (CEUs)
Copyright © 2007-2013 EMLab P&K. All rights reserved. 58
Overview of Health Effects of Bacteria
• Infection
• Allergy
• Toxicosis
• Symbiosis
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Infection
Invasion by, and multiplication of, “pathogenic”
microorganisms in a bodily part or tissue, which may
produce subsequent tissue injury and progress to overt
disease through a variety of cellular or toxic mechanisms
Overwhelming communities of bacteria
Immune reactions leading to disease
Release of toxins that cause the symptoms of disease.
Copyright © 2007-2013 EMLab P&K. All rights reserved. 60
The Term “Pathogen”
• A pathogen is defined as:
– A microorganism able to cause infection
• Limited to infectious agents
— OR —
– A microorganism able to cause disease
• Includes humidifier fever, hypersensitivity pneumonitis,
allergy (rarely), toxicoses
• Obligate pathogens
– Only grow in the host; not recovered from the environment
• Environmental pathogens
– Pathogens that grow naturally in the environment but also
cause disease when they invade host tissue
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Virulence
• Virulent infection: infection by an organism that can
infect anyone without specific antibodies.
• Virulence: a measure of how readily an organism can
infect a host.
– Highly virulent agents may cause infection following exposure
to a single organism. (e.g., Mycobacterium tuberculosis)
– Less virulent organisms may require higher doses before
infection occurs (e.g., Bacillus anthracis)
– Avirulent organisms may only infect those without a
functioning immune system (e.g, Pseudomonas aeruginosa)
• Opportunistic infection
– Caused by avirulent organisms
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Risk Factors for Infection
• Immune status
– Lack of induced specific antibodies
– Age. Neonates and the elderly are at increased risk of bacterial infections.
– Nutritional status. Malnutrition results in a depressed immune system
– Genetic predisposition. The Human Genome Project increased our ability to locate specific genes related to infectious disease susceptibility ( Bentley DR 2000).
– Immunosuppression via disease or medications
Copyright © 2007-2013 EMLab P&K. All rights reserved. 63
Risk Factors for Infection: Exposure
• Amount of exposure
– Depends on virulence of the organism
– Depends on immune status of the host
• Exposure pathway
– Depends on the organism
• Mycobacterium tuberculosis: must be inhaled into lower airway
• GI tract agents must be ingested
• Exogenous: those that originate outside the body
– Food, water, air, surfaces, other people
– Ingestion, inhalation, other entry points.
• Endogenous: caused by bacteria within the body that cause disease when the body's resistance is lowered
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Pathways To Infection
• Airborne infection – inhalation of microorganisms or spores suspended in air, on dust
particles, or in droplet nuclei (dried droplets).
• Droplet infection – inhalation of respiratory pathogens suspended for a brief time on
liquid particles exhaled by someone already infected.
• Direct contact – Touching an infected person or contaminated surface
• Endogenous infection – reactivation of organisms present in a dormant focus, as occurs
in tuberculosis, etc.
• Tunnel infection – subcutaneous infection of an artificial passage into the body that
has been kept open.
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Respiratory Infections
• Respiratory infections involve the nose, throat, and all the airway passages plus the ears.
• Common respiratory infections
– Colds and flu (viral)
– Sinus infections and ear infections
– Bronchitis
– Pneumonia
• Modes of infection
– Airborne
– Droplets
– Direct contact
– Endogenous
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URI and Ear Ache
• Upper respiratory tract infections (URI)
– Leading cause of time lost from work and school
– Bacteria account for up to 25 percent of URI (the rest are viral)
– Streptococcus, Haemophilus influenzae
• Otitis media
– Middle ear infection: most common bacterial infection in U.S. children.
– Streptococcus pneumoniae
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Tuberculosis
• Tuberculosis (TB)
– Mycobacterium tuberculosis
– Affects 15 million people in the U.S. Fewer develop disease which depends on nutritional status, age, HIV, incarceration
– Major cause of death in many countries; in some cases virtually all of the population is infected and a large proportion develop disease
– Organism must be inhaled into the lower airway.
– Colonization occurs, causing development of antibodies detectable with skin test
– Active disease occurs when some change occurs in the immune system
– Treatment is long term and compliance is often poor
– Control requires prevention of exposure to patients with active disease
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Legionnaires’ Disease
Legionella
• Gram negative bacteria
common in many
environments
• Approx. 50 species and 70
serogroups have been
described
• Legionella is the causative
agent of Legionellosis
(Legionnaires’ disease
and Pontiac fever)
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Legionnaires’ Disease (cont’d)
Legionnaires’ Disease
Legionellosis takes two distinct forms:
1) Pontiac fever: respiratory illness without pneumonia, symptoms
resemble acute influenza (not a lower respiratory illness)
2) Legionnaires’ disease: symptoms include fever, chills, cough, muscle
aches, headache, tiredness, loss of appetite, loss of coordination (ataxia),
and occasionally diarrhea and vomiting.
• Community and Hospital Acquired
• Risk factors: Exposure, compromised immune function
– Transmission is not person to person
– Age: Highest risk in elderly >65; uncommon in people <50; very rare in people <20
– Smoking
– Pre-existing chronic obstructive pulmonary disease (COPD), diabetes
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Infection and Transmission
• Infection occurs after inhaling water droplets that originated from a water source contaminated with Legionella.
• Manmade water sources include cooling towers, evaporative coolers, hot water systems, showers, whirlpool spas, architectural fountains, room-air humidifiers, ice-making machines, misting equipment.
• Environmental sources for Legionella are freshwater ponds, rivers and creeks.
• Legionella survives in the environment as intracellular parasites of freshwater protozoa.
• Legionella bacteria will grow in water at temperatures 20C to
50C (68F to 122F).
• Ideal growth conditions are in stagnant water (95F to 115F)
Legionnaires’ Disease (cont’d)
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Legionnaires’ Disease (cont’d)
• Threshold limits for
potable water
especially in hospitals
and nursing homes
should be considerably
lower.
• Goal for “sensitive
locations” is a zero
count for Legionella
(detection limits are
typically around 100
cfu/liter)
Copyright © 2007-2013 EMLab P&K. All rights reserved. 72
Sampling for Legionella
• Wear Respiratory Protection: Wear appropriate
respiratory protection during the examination of water
systems if a significant potential exists for exposure to
high concentrations of contaminated aerosols (e.g.
operating spray humidifier).
• Swabs: Sampling of biofilm (slime) or on water outlets
(e.g. inside of shower heads). Use sterile swab and keep
moist.
• Air: Air samples collected on special culture plates with
an Andersen-type sampler rarely demonstrate the
presence of Legionella in the air. Not recommended.
Copyright © 2007-2013 EMLab P&K. All rights reserved. 73
Specific Guidelines for Legionella
Water samples:
• Non-potable water source (e.g. cooling towers, chillers, condensate pans, surface water in reservoirs, sprinklers): Collect 250 ml water from the bottom or side of the vessel or reservoir.
• Potable water source:
– Use 1 liter bottles containing thiosulfate to neutralize chlorine.
– Collect a 250-mL to 1-Liter “pre-flush" sample of the first water drawn from bottom drains and outlet valves of storage tanks, sumps, and water heaters as will as faucets and showerheads.
– Allow the water to run until the temperature stabilizes and collect a second "post-flush" sample when the water temperature is constant (after approx. 60 sec.).
Copyright © 2007-2013 EMLab P&K. All rights reserved. 74
Water Treatment
Water treatment options to eradicate Legionella
• Thermal Eradication
• Copper-Silver Ionization (ionization unit)-best long term treatment
• Chlorination
• Ozonation
• Chlorine Dioxide
• Ultraviolet Irradiation (point of delivery treatment)
Heat treatment:
70 to 80C (158 to 176F): Disinfection range
At 66C (151F): Legionellae die within 2 minutes
At 60C (140F): Legionellae die within 32 minutes
At 55C (131F): Legionellae die within 5 to 6 hours
Copyright © 2007-2013 EMLab P&K. All rights reserved. 75
Legionella – Thresholds
CFU of Legionella
per Liter Action
< 1000 System under control
1000 – 10,000 Review program operation. Conduct re-sampling.
Review of control measures and risk assessment
should be carried out to identify any remedial
actions.
> 10,000 Implement corrective action. The system should
immediately be re-sampled.
• No concrete threshold and action limits for Legionella:
• The European Working Group for Legionella Infections
(EWGLI) published the following guidelines and action
limits for cooling towers:
Copyright © 2007-2013 EMLab P&K. All rights reserved. 76
Legionella – More Information
More information and literature can be found at:
• http://en.wikipedia.org/wiki/Legionnaire%27s_Disease
• http://www.cdc.gov/legionella/index.htm
• http://www.ewgli.org/
Copyright © 2007-2013 EMLab P&K. All rights reserved. 77
Inhalation Anthrax
• Mostly confined to
handlers of infected
animals
• Most of these develop
skin infection
• Inhalation infection rare
• Biowarfare problem
Copyright © 2007-2013 EMLab P&K. All rights reserved. 78
Gastrointestinal Infections
• GI infections involve the stomach, intestines, and colon
– Gastroenteritis: nausea, vomiting, diarrhea, abdominal
discomfort
– Diarrhea: abnormal fecal discharge with frequent and/or
fluid stool; usually resulting from disease of the small
intestine and resulting in fluid loss and dehydration
– Dysentery: inflammatory disorder accompanied by blood
and pus in the feces, abdominal pain, fever, and cramps;
usually results from disease of the large intestine.
– Enterocolitis: inflammatory disorder of both the small and
large intestine.
Copyright © 2007-2013 EMLab P&K. All rights reserved. 79
Common GI Infections
• Shiga toxin-producing E. coli (E. coli O157:H7 and other
serotypes)
• Salmonella species other than S. Typhi (salmonellosis)
• Shigella species (shigellosis)
• Campylobacter species
• Clostridum difficile and antibiotic-associated colitis
Copyright © 2007-2013 EMLab P&K. All rights reserved. 80
Shiga Toxin-producing E. Coli
(E. Coli O157:H7 and Other Serotypes)
• Estimated 20,000 cases of diarrheal disease each year.
• Most human infections transmitted by beef resulting from fecal
contamination of meat during slaughter. Undercooked hamburger
meat is a common problem. Other reported sources:
– Unchlorinated drinking water
– Unpasteurized apple cider
– Alfalfa sprouts
– Leaf lettuce
– Mesclun lettuce
– Radish sprouts
– Milk
• Infectious dose is low, possibly as low as 100 bacteria.
Therefore, prevention is difficult and person-to-person spread
may occur.
Copyright © 2007-2013 EMLab P&K. All rights reserved. 81
Salmonella Species Other Than
S. Typhi (Salmonellosis)
• Estimated 0.8 to 3.7 million cases each year.
• Human disease is usually associated with food products,
especially poultry and eggs. Other reported sources:
– Tomatoes
– Alfalfa sprouts
– Cantaloupe
– Freshly squeezed orange juice
– Keeping exotic pets, especially reptiles, of which up to
90% harbor Salmonella
Copyright © 2007-2013 EMLab P&K. All rights reserved. 82
Shigella Species (Shigellosis)
• Classic agents of dysentery
• At least 19,000 cases occur in the U.S. each year
• Shigella can contaminate food and water.
• Person-to-person transmission is the dominant mode of
transmission.
• As few as 10 to 100 Shigella organisms can cause
disease; therefore, shigellosis is highly communicable.
• Outbreaks of shigellosis occur in crowded, closed
environments such as nurseries, day care centers,
institutions, and cruise ships.
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Campylobacter Species
• One of the most common infectious diseases worldwide
• More than 1 million cases occur each year in the U.S.
• Human disease is mostly food-borne but can result from
direct contact with animals, including household pets
(notably, puppies or kittens with diarrhea)
• Fecal-oral transmission occurs.
• Transmission from food handlers appears to be
uncommon.
• Disease can result from ingestion of as few as 500
bacilli.
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Clostridum Difficile and Antibiotic-
associated Colitis
• Diarrhea is a relatively common complication of
antimicrobial therapy and is associated with Clostridium
difficile in about 10% to 30% of cases
• About 60% of adults in the U.S. have serum antibodies
to C. difficile
• Person-to-person transmission of C. difficile occurs, and
can be reduced by hand washing before and after
patient encounters and by use of disposable gloves
(followed by hand washing) during direct contact with
patients.
Copyright © 2007-2013 EMLab P&K. All rights reserved. 85
Sewage
• Because of its nature, GI tract disease is the most likely outcome of sewage exposure.
• Other diseases transmitted by sewage are not bacteria (Hepatitis, for example).
• Bacteria are used as indicators for sewage exposure.
Copyright © 2007-2013 EMLab P&K. All rights reserved. 86
What Are The Health Risks?
• Gastroenteritis: cramping stomach pains, diarrhea and
vomiting
• Weil's disease (leptospirosis) animal source bacterial
infection: a flu-like illness with persistent and severe headache;
damage to liver, kidneys and blood may occur and the condition
can be fatal (Leptospira)
• Hepatitis: inflammation of the liver, and jaundice (viral)
• Occupational asthma: attacks of breathlessness, chest
tightness and wheezing; produced by the inhalation of living or
dead organisms
• Infection of skin or eyes
• Rarely, allergic alveolitis: (inflammation of the lung) with
fever, breathlessness, dry cough, and aching muscles and joints
Copyright © 2007-2013 EMLab P&K. All rights reserved. 87
Who Is At Risk?
• Employees involved in sewer inspection and maintenance work
• Construction workers who repair or replace live sewers
• Water company employees who work with sewage treatment plants
• Agricultural and forestry workers who may be exposed to sewage sludge
• Sludge tanker drivers/operators and associated maintenance staff
• Plumbers
• Employees who clean and maintain the underside of railway carriages and empty aircraft sewage compartments and other types of portable lavatories.
Copyright © 2007-2013 EMLab P&K. All rights reserved. 88
How Do Sewage Micro-organisms Enter
The Body?
• Hand-to-mouth contact during eating, drinking and smoking,
• Wiping the face with contaminated hands or gloves, or by licking splashes from the skin.
• Skin contact, through cuts, scratches, or penetrating wounds, i.e. from discarded hypodermic needles.
• Aerosols landing on surfaces of the eyes, nose and mouth.
• By breathing them in, as either dust, aerosol or mist.
Copyright © 2007-2013 EMLab P&K. All rights reserved. 89
Aerosol Exposure
• Wastewater treatment
– Average concentrations of 17000 cfu/ml3 of mesophilic,
2100 cfu/ml3 of TSA-SB bacteria (bacteria associated with
certain waterborne virulence factors) in the water.
– In the aerosol of the fixed-film reactor 3000 cfu/m3
mesophilic and 730 cfu/m3 TSA-SB bacteria.
Copyright © 2007-2013 EMLab P&K. All rights reserved. 90
Coliforms
• Rod-shaped Gram-negative non-spore forming organisms that ferment lactose with the production of acid and gas when incubated at 35-37°C.
• Coliforms are abundant in the feces of warm-blooded animals, but can also be found in the aquatic environment, in soil and on vegetation
• TAXA – Citrobacter
– Enterobacter
– Escherichia
– Hafnia
– Klebsiella
– Serratia
– Yersinia
Copyright © 2007-2013 EMLab P&K. All rights reserved. 91
Water Quality Coliform Guidelines
• Culture based
– 200 colonies of fecal coliform bacteria /100ml for primary
contact recreation
– 1000 colonies of fecal coliform bacteria /100ml for
secondary contact recreation
• Swimming beaches:
– Geometric mean of 126 E.coli bacteria per 100 ml of water
(fresh water)
Copyright © 2007-2013 EMLab P&K. All rights reserved. 92
Enterococcus
• Higher correlation than fecal
coliforms with many human
pathogens
• In 2004, Enterococcus spp.
became the new federal
standard for water quality at
public beaches
• The acceptable level of
contamination is very low,
– Hawaii: 7 colony forming units
per 100 ml of water
– Geometric mean of 35 /100 ml
of water for five samples over
30 days and an instantaneous
(single sample) standard of 104
/ 100 ml of water (salt water).
Enterococcus infection in pulmonary tissue.
Source: wikipedia.org
Copyright © 2007-2013 EMLab P&K. All rights reserved. 93
• Remove all liquid and solid material as quickly as
possible.
• Residual organisms will stick to the dry surface and
eventually die (by orders of magnitude per hour).
• As a final precaution, soapy water with ~ 1/4 -1/2 cup
bleach/gallon can be used.
• Use skin protection especially over cuts and scrapes.
• Do not ever touch the face. Remember, the outside of
gloves are contaminated!
Sewage Spill Cleanup
Copyright © 2007-2013 EMLab P&K. All rights reserved. 94
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
Copyright © 2007-2013 EMLab P&K. All rights reserved. 95
MRSA and Staphylococci – Overview
Staphylococcus aureus
• Gram-positive bacteria
• 0.5 – 1 m diameter
• Commonly found on the
skin and in the nose of
humans
Copyright © 2007-2013 EMLab P&K. All rights reserved. 96
MRSA – Resistance
• MRSA is a resistant variation of the common bacterium
Staphylococcus aureus. It has evolved an ability to
survive treatment with beta-lactam antibiotics, including
penicillin, methicillin, and cephalosporins
– Hospital acquired (HA)
– Community acquired (CA)
Copyright © 2007-2013 EMLab P&K. All rights reserved. 97
• Approximately one third of the world’s population has S.
aureus bacteria on their body at any given time
• About 1% of people carrying S. aureus have MRSA
(CDC estimated)
• Spread from one person to another through casual
contact or contaminated objects
• Estimated 94,000 cases of MRSA infections in the U.S.
per year and nearly 19,000 deaths.
MRSA and Staphylococci – Overview
(cont’d)
Copyright © 2007-2013 EMLab P&K. All rights reserved. 98
Staphylococci – Symptoms
Staphylococcus
• Folliculitis (hair roots)
• Impetigo (blisters)
• Skin abscesses
• Cellulites
• Necrolysis
Copyright © 2007-2013 EMLab P&K. All rights reserved. 99
S. aureus – Systemic Infections
• Staphylococci tend to infect skin but can travel through the bloodstream and involve almost any site in the body, particularly the heart and the bones
• May infect the respiratory tract, mainly in people with chronic lung disease or influenza and lead to staphylococcal pneumonia
• Other severe and life-threatening infections with staphylococci include infections of the heart and victims of severe burns
Copyright © 2007-2013 EMLab P&K. All rights reserved. 100
Hospital Acquired MRSA
• Most infections with MRSA occur in hospitals and
healthcare facilities (HA-MRSA) including:
– Surgical wound infections
– Urinary tract infections
– Blood stream infections and pneumonia
Copyright © 2007-2013 EMLab P&K. All rights reserved. 101
HA-MRSA – Risk Factors and Prevention
Risk factors for HA-MRSA:
• Current or recent hospitalization
• Residing in a long-term care facility
• Invasive devices
• Recent antibiotic use
Prevention:
• Ask hospital staff to wash their hands or use gloves
• Wash your own hands frequently
• Make sure invasive devices are kept sterile
Copyright © 2007-2013 EMLab P&K. All rights reserved. 102
Community Acquired MRSA
Community acquired MRSA recently gained much attention in the news media.
Risk factors:
• Young age (children)
• Participating in contact sports
• Sharing towels and athletic equipment
• Weakened immune system
• Living in crowded and/or unsanitary conditions
• Association with health care workers
Copyright © 2007-2013 EMLab P&K. All rights reserved. 103
Community Acquired MRSA
Prevention
• Washing hands frequently
• Keep personal items personal
• Keep wounds covered
• Shower after athletic games or practices
• Do not participate in athletic games or practices if you have infected wounds
• Sanitize items
• Get tested if you have a skin infection
• Use antibiotics appropriately
Copyright © 2007-2013 EMLab P&K. All rights reserved. 104
Diagnosis and Environmental Testing
Clinical testing
• Sample from an infected site or a nasal swab
• Clinical laboratory performs the testing
Environmental testing
• Collect swab samples from items that are frequently touched such as door knobs, keyboards, athletic equipment etc.
• Environmental laboratory can perform the testing for MRSA
Copyright © 2007-2013 EMLab P&K. All rights reserved. 105
Allergy
• IgE allergy
– Most patients with nasal polyposis and/or chronic
sinusitis possess bacteria-specific IgE, while subjects
with only allergic rhinitis do not;
– Multiple bacterial species isolated from chronically
infected sinuses are capable of inducing IgE-
mediated sensitization. (Calenoff et al 1993)
• IgG/cell mediated allergy
– Machining coolant aerosols?
– Thermophilic actinomycetes
Copyright © 2007-2013 EMLab P&K. All rights reserved. 106
Bacterial Toxicoses
General definitions
• Endotoxins: Toxic compounds
found in bacteria and other
pathogens.
• Exotoxins: Secreted in soluble
form.
Copyright © 2007-2013 EMLab P&K. All rights reserved. 107
Toxicoses: Endotoxin
Endotoxins
• Component of cell walls
of gram-negative
bacteria (E. coli,
Salmonella etc. )
• Lipopolysaccharide
(LPS) - Potent
stimulator of the
immune system
Detection with Limulus
Amebocyte Lysate
Copyright © 2007-2013 EMLab P&K. All rights reserved. 108
• Clinical effects described at 50 EU/m3 include:
– General symptoms of fever, shivering, joint pain and fatigue
(most common).
– Respiratory effects include dyspnea, chest tightness,
coughing, non-allergic asthma and reduction in lung function
• Higher sensitivity in people with pre-existing respiratory
disease.
• Relationship between endotoxin and Asthma is unclear:
Endotoxins may protect against allergic asthma but is a
risk factor for non-allergic asthma.
• Smoking, presence of furred pets and cleaning regime
are correlated with endotoxin levels.
Endotoxins (continued)
Copyright © 2007-2013 EMLab P&K. All rights reserved. 109
Endotoxins – Sampling
Sampling:
• Preferred sample type is air (endotoxin-free filter
cassettes)
• 250 – 1000 Liter sample volume for air
• Dust can also be used as sample type
• Metal working fluids, few milliliter sample
Copyright © 2007-2013 EMLab P&K. All rights reserved. 110
• Recommendation of ~100 EU/m3 as maximum exposure
limit. Background levels of >10 EU/m3 are of concern
and >30 EU/m3 should be avoided.
• Normal concentrations indoors: <1 EU/m3
• Higher concentrations may indicate water damage.
• Concentrations increase up to 100-fold in rooms of
smokers.
Endotoxins – Exposure Limits
Copyright © 2007-2013 EMLab P&K. All rights reserved. 111
Endotoxins – Control and Prevention
• Most serious illness occurs in industrial environments
where ventilation controls are the primary approach
• Controlling water reduces possibility of Gram-negative
bacteria (and endotoxin)
• Removal of contaminated materials and HEPA
vacuuming can reduce endotoxin levels
• Do not smoke
Copyright © 2007-2013 EMLab P&K. All rights reserved. 112
Toxicoses: Exotoxins
Toxin Toxic Dose
(mg) Host
Compared to
Strychnine
Botulinum
toxin 0.8x10-8 Mouse 3x106
Tetanus toxin 4x10-8 Mouse 1x106
Shiga toxin 2.3x10-6 Rabbit 1x106
Diphtheria
toxin 6x10-5 Guinea pig 2x103
Copyright © 2007-2013 EMLab P&K. All rights reserved. 113
"If we have not gotten our message across, then we ought to
assume that the fault is not with our receivers."
Baruch Fischhoff, Department of Engineering and Public Policy
Carnegie-Mellon University (1985)
Risk Communication
Risk Communication:
An interactive process of exchange of information and
opinion among individuals, groups, and institutions; often
involves multiple messages about the nature of risk or
expressing concerns, opinions, or reactions to risk
messages or to legal and institutional arrangements for
risk management.
Copyright © 2007-2013 EMLab P&K. All rights reserved. 114
Risk Communication (continued)
• Risk communication is sometimes an
important element of an investigation
• Important elements to successful
communication are establishing,
maintaining, and increasing trust and
credibility
• From Dr. Covello at Columbia Univ.
Center for Risk Commun.: Important
elements of trust and credibility are:
– Caring & empathy
– Openness & honesty
– Commitment & dedication
– Expertise & competence
Copyright © 2007-2013 EMLab P&K. All rights reserved. 115
Risks perceived:
• To be voluntary are more accepted than risks perceived to be imposed
• To be fairly distributed are more accepted than risks perceived to be
unfairly distributed.
• To be natural are more accepted than risks perceived to be manmade.
• To be statistical are more accepted than risks perceived to be
catastrophic.
• To be generated by a trusted source are more accepted than risks
perceived to be generated by an untrusted source.
• To be familiar are more accepted than risks perceived to be exotic.
• To affect adults are more accepted than risks perceived to affect
children.
Some Factors Influencing Risk
Perception (Fischhoff et al. 1981)
Copyright © 2007-2013 EMLab P&K. All rights reserved. 116
• The cardinal rule of risk communication is: first do no
harm.
• Gain a sense of the general attitude toward the situation
and tailor your presentation accordingly.
– Are they worried and in need of reassurance?
– Are they sanguine and in need of a warning?
– Are they angry and in need of calming?
Rules and Tips
Copyright © 2007-2013 EMLab P&K. All rights reserved. 117
• Review your remarks to gauge the probable impact that
your words will have on the situation and adjust them as
necessary; e.g.
• Are you using words, like “crisis,” “life-threatening,” or
“extremely”
• Can other, less dramatic words be substituted?
Rules and Tips (continued)
Copyright © 2007-2013 EMLab P&K. All rights reserved. 118
• Don’t speak unless you are comfortable doing so.
• Most communications mistakes are made by those who
are not prepared to speak but feel compelled to do so
anyway.
• If confronted with a demand for a quick answer, have the
confidence to say, “I would like to answer that question
later.”
Rules and Tips (continued)
Copyright © 2007-2013 EMLab P&K. All rights reserved. 119
• Informing clients (or the public) of the problem and
specific dangers, providing guidance on appropriate
responses, and easing concerns are achievable goals.
• If the risk is low, say: “The risk to the public is low.”
• Don’t elaborate with scientific jargon including statistics.
Rules and Tips (continued)
Copyright © 2007-2013 EMLab P&K. All rights reserved. 120
• Remember the rules for risk communication; don’t write
inflammatory statements.
• Write a straightforward factual account of your
interpretation of the data you have collected.
• Don’t make health inferences unless you are a qualified
health professional.
• If you don’t do remediation work, don’t recommend
remediation methods.
• Don’t fill the report with boilerplate
Communicating In Written Reports
Copyright © 2007-2013 EMLab P&K. All rights reserved. 121
A Few Risk Communication Resources
Web sites
Agency for Toxic Substances & Disease
Registry:
Primer on Health Risk Communication: http://www.atsdr.cdc.gov/HEC/primer.html
National Institute of Corrections:
Risk Communication Guidelines for
Public Officials:
http://nicic.org/Library/019092
Classes
The Centers for Disease Control offers
several classes
Copyright © 2007-2013 EMLab P&K. All rights reserved. 122
To receive a certificate of attendance, you must complete the survey after the webinar:
• Click on the survey link in the “Thank you” email (sent 1 hour after this webinar).
• Complete survey by this Friday, March 15, 2013.
• You will receive an email in 2-3 weeks with instructions when your certificate is ready.
Continuing Education Units (CEUs)
Copyright © 2007-2013 EMLab P&K. All rights reserved. 123
Thank you for your time!
Questions about Allergens or Bacteria: DGallup@emlabpk.com
All other questions: webinars@emlabpk.com
Copyright © 2007-2013 EMLab P&K. All rights reserved. 125
Locations Nationwide to Serve You (Addresses on Following Slides)
WASHINGTON
OREGON
IDAHO
NEVADA
MONTANA
WYOMING
UTAH
ARIZONA
COLORADO
NEW
MEXICO
TEXAS
NORTH
DAKOTA
SOUTH
DAKOTA
NEBRASKA
KANSAS
OKLAHOMA
MINNESOTA
IOWA
MISSOURI
ARKANSAS
LOUISIANA
WISCONSIN
ILLINOIS INDIANA OHIO
KENTUCKY
TENNESSEE
GEORGIA
SOUTH
CAROLINA
NORTH
CAROLINA
VIRGINIA
NEW YORK
PENNSYLVANIA
MICHIGAN
MAINE
WEST
VIRGINIA
VERMONT
NEW
HAMPSHIRE
MARYLAND
DELAWARE
NEW JERSEY
CONNECTICUT
RHODE ISLAND
MASSACHUSETTS
HAWAII
Copyright © 2007-2013 EMLab P&K. All rights reserved. 126
EMLab P&K Locations Near You
MicroLabs in bold are
AIHA Accredited as
documented by the
Scope of Accreditation
Certificate.
Arizona - Phoenix
1501 West Knudsen Drive
Phoenix, AZ 85027
phone: 800.651.4802
fax: 623.780.7695
AIHA EMLAP# 102297
California - San Bruno
1150 Bayhill Drive, Suite 100
San Bruno CA 94066
phone: 866.888.6653
fax: 650.829.5852
AIHA EMLAP# 102856
New Jersey - Cherry Hill
1936 Olney Avenue
Cherry Hill, NJ 08003
phone: 866.871.1984
fax: 856.489.4085
AIHA EMLAP #103005
M I C R O L A B S
L A B O R A T O R I E S
California - San Diego
9089 Clairemont Mesa Blvd.
Suite 106
San Diego, CA 92123
Phone: 866.465.6653
Colorado - Denver
4955 Yarrow Street
Arvada, CO 80002
phone: 800.651.4802
Florida - Ft. Lauderdale
6301 NW 5th Way
Suite 2850
Ft. Lauderdale, FL 33309
phone: 877.711.8400
Illinois - Chicago
655 W. Grand Ave.
Suite 205
Elmhurst, IL 60126
phone: 866.871.1984
Nevada - Las Vegas
6000 S. Eastern Ave.
Suite 5E
Las Vegas, NV 89119
phone: 866.888.6653
Texas - Houston
6310 Rothway St.
Houston, Texas 77040
phone: 800.651.4802
California - Glendale
1010 N. Central Ave.
Suite 420
Glendale, CA 91202
phone: 866.465.6653
California - Irvine
17461 Derian Ave
Suite 100
Irvine, CA 92614
Phone: 866.465.6653
California - Sacramento
880 Riverside Parkway
West Sacramento, CA 95605
phone: 866.888.6653
Virginia - Fairfax
3929 Old Lee Highway
Unit 91C
Fairfax, Virginia 22030
phone: 866.871.1984
Washington - Seattle
19515 North Creek
Parkway N, Suite 100
Bothell, WA 98011
phone: 866.888.6653
For the most current list of locations, please visit us at www.emlabpk.com
Contact individual laboratories for service capabilities and scopes of accreditation.
Copyright © 2007-2013 EMLab P&K. All rights reserved. 127
TestAmerica Locations
For 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 - Pleasanton
1220 Quarry Ln.
Pleasanton, CA 94566
phone: (925) 484-1919
CALIFORNIA - San Bernardino
202 E. Airport Road
Suite 140
San Bernardino, CA 92408
Phone: (909) 370-4707
CALIFORNIA – W. Sacramento
880 Riverside Pkwy
West Sacramento, CA 95605
phone: (916) 373-5600
CONNECTICUT
128 Long Hill Cross Rd.
Shelton, CT 06484
phone: (203) 929-8140
FLORIDA - Jacksonville
8933 Western Way, Suite 1
Jacksonville, FL 32256
phone: (904) 519-9551
FLORIDA - Orlando
8010 Sunport Drive, Suite 116
Orlando, FL 32809
phone: (407) 851-2560
FLORIDA - Pensacola
3355 McLemore Dr.
Pensacola, FL 32514
phone: (850) 474-1001
FLORIDA - Tallahassee
2846 Industrial Plaza Dr.
Tallahassee, FL 32301
phone: (850) 878-3994
FLORIDA - Tampa
6712 Benjamin Rd., Suite 100
Tampa, FL 33634
phone: (813) 885-7427
GEORGIA - Atlanta
6500 McDonough Drive, Suite C-10
Norcross, GA 30093
phone: (678) 966-9991
GEORGIA - Savannah
5102 LaRoche Avenue
Savannah, GA 31404
phone: (912) 354-7858
HAWAII - Honolulu
99-193 Aiea Heights Dr.
Suite 121
Aiea, HI 96701
phone: (808) 486-5227
ILLINOIS - Chicago
2417 Bond Street
University Park, IL 60484
phone: (708) 534-5200
ILLINOIS - Elmhurst
655 W. Grand Ave., Suite 205
Elmhurst, IL 60126
phone: (630) 758-0262
INDIANA - Indianapolis
Stutz Business Center
212 W. 10th Street, Ste A-205
Indianapolis, IN 46202
Phone: (317) 264-9686
INDIANA - Valparaiso
2400 Cumberland Drive
Valparaiso, IN 46383
phone: (219) 464-2389
IOWA - Cedar Falls
704 Enterprise Drive
Cedar Falls, IA 50613
phone: (319) 277-2401
IOWA - Davenport
736 Federal St., Suite 2202
Davenport, IA 52803
phone: (563) 323-7944
LOUISIANA - Baton Rouge
6113 Benefit Dr.
Baton Rouge, LA 70809
phone: (225) 755-8200
MARYLAND - Baltimore
5710 Executive Drive, Suite 106
Baltimore, MD 21228
phone: (410) 869-0085
MASSACHUSETTS - Boston
240 Bear Hill Rd., Suite 104
Waltham, MA 02451
phone: (781) 466-6900
MASSACHUSETTS - Westfield
53 Southampton Road
Westfield, MA 01085
phone: (413) 572-4000
MICHIGAN - Brighton
10448 Citation Drive, Suite 200
Brighton, MI 48116
Phone: (810) 229-2763
MINNESOTA - Minneapolis
7204 West 27th Street, Suite 114
St. Louis Park, MN 55426
phone: (800) 593-8519
ALABAMA - Mobile
900 Lakeside Drive
Mobile, AL 36693
phone: (251) 666-6633
ALASKA - Anchorage
2000 W. International Airport
Rd., Suite A10
Anchorage, AK 99502
phone: (907) 563-9200
ARIZONA - Phoenix
4625 E. Cotton Center Blvd.,
Suite 189
Phoenix, AZ 85040
phone: (602) 437-3340
ARIZONA - Tucson
1870 W. Prince Road, Suite 59
Tucson, AZ 85705
phone: (520) 807-3801
CALIFORNIA – Costa Mesa
3585 Cadillac Ave, Suite A
Costa Mesa, CA 92626
phone: (714) 258-8610
Copyright © 2007-2013 EMLab P&K. All rights reserved. 128
TestAmerica Locations (cont’d)
NEW YORK - Albany
25 Kraft Ave.
Albany, NY 12205
phone: (518) 438-8140
NEW YORK - Buffalo
10 Hazelwood Drive, Ste. 106
Amherst, NY 14228
phone: (716) 691-2600
NEW YORK - New York City
47-32 32nd Place, Suite 1141
Long Island City, NY 11101
Phone: (347) 507-0579
NEW YORK - Syracuse
118 Boss Rd.
Syracuse, NY 13211
phone: (315) 431-0171
NORTH CAROLINA - Charlotte
I-85 South Bldg.
2858 Queen City Dr., Suite B
Charlotte, NC 28208
phone: (704) 392-1164
NORTH CAROLINA - Raleigh
101-F Woodwinds Industrial Court
Cary, NC 27511
phone: (919) 380-9919
For 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.
OHIO - Cincinnati
11416 Reading Road
Cincinnati, OH 45241
phone: (513) 733-5700
OHIO - Dayton
4738 Gateway Circle
Dayton, OH 45440
Phone: (937) 294-6856
OHIO - North Canton
4101 Shuffel Street NW
North Canton, OH 44720
phone: (330) 497-9396
OREGON - Portland
9405 SW Nimbus Avenue
Beaverton, OR 97008
phone: (503) 906-9200
PENNSYLVANIA - King of Prussia
1008 W. Ninth Ave.
King of Prussia, PA 19406
phone: (610) 337-9992
PENNSYLVANIA - Pittsburgh
301 Alpha Drive
Pittsburgh, PA 15238
phone: (412) 963-7058
SOUTH CAROLINA - Charleston
1436-A North Point Lane
Mt. Pleasant, SC 29464
phone: (843) 849-6550
TENNESSEE - Knoxville
5815 Middlebrook Pike
Knoxville, TN 37921
phone: (865) 291-3000
TENNESSEE - Nashville
2960 Foster Creighton Dr.
Nashville, TN 37204
phone: (615) 726-0177
TEXAS - Austin
14050 Summit Dr., Ste. A100
Austin, TX 78728
phone: (512) 244-0855
TEXAS - Beaumont
4400 Lawndale Ave.
Groves, TX 77619
phone: (409) 540-5302
TEXAS - Corpus Christi
1733 N. Padre Island Drive
Corpus Christi, TX 78408
phone: (361) 289-2673
MISSOURI - Eureka
1699 West Fifth Street, #200
Eureka, MO 63025
Phone: (314) 302-8354
MISSOURI - Kansas City
601 NW 39th Street
Blue Springs, MO 64015
phone: (800) 276-1286
MISSOURI - St. Louis
13715 Rider Trail North
Earth City, MO 63045
phone: (314) 298-8566
NEW JERSEY - Edison
777 New Durham Road
Edison, NJ 08817
phone: (732) 549-3900
NEW JERSEY - South Jersey
520 Fellowship Rd., Suite A-106
Mt. Laurel, NJ 08054
phone: (856) 222-1990
TEXAS - San Antonio
404 E. Ramsey, Suite 208
San Antonio, TX 78216
phone: (210) 344-9751
VERMONT - Burlington
30 Community Drive, Suite 11
South Burlington, VT 05403
phone: (802) 660-1990
VIRGINIA - Virginia Beach
5135 Cleveland Street
Virginia Beach, VA 23462
phone: (757) 671-1291
WASHINGTON - Richland
2800 George Washington Way
Richland, WA 99354
phone: (509) 375-3131
WASHINGTON - Spokane
11922 E. 1st Ave.
Spokane, WA 99206
phone: (509) 924-9200
WASHINGTON - Tacoma
5755 8th Street East
Tacoma, WA 98424
phone: (253) 922-2310
Copyright © 2007-2013 EMLab P&K. All rights reserved. 129
EMLab P&K Products
Authorized Distributor for:
Buy equipment and supplies for sampling
allergens, asbestos, bacteria, mold, fungi, and more
Shop online at www.emlabpk.com/store
Copyright © 2007-2013 EMLab P&K. All rights reserved. 130
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
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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 to
evaluate 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 of
the 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)
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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.
EMLab P&K General Glossary (continued)
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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 EMLab
P&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.
EMLab P&K General Glossary (continued)
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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.
EMLab P&K General Glossary (continued)
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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
EMLab P&K General Glossary (continued)
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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.
EMLab P&K General Glossary (continued)
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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 measurement
system 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.
EMLab P&K General Glossary (continued)
Copyright © 2007-2013 EMLab P&K. All rights reserved. 138
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.
EMLab P&K General Glossary (continued)
Copyright © 2007-2013 EMLab P&K. All rights reserved. 139
When quality and accuracy are critical. Analytical Services: Allergens, Asbestos, Bacteria, Fungi, PCR, Radon, & USP <797>
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