We
welcome our noted banquet speaker...
Shellfish
and the Treaty Tribes of Washington State:
Fisheries History-in-the-Making
David
Fyfe, Shellfish Biologist, Northwest Indian
Fisheries Commission.
By the mid 1800's, the US government and local
tribes had concluded that treaties were preferable
to war. In an effort to protect as many of everyone's
interests as possible, 5 treaties were negotiated
in 1854 & 1855. Washington was not yet a
state, but the treaties applied to the area
that is now western Washington. In the years
that followed, much growth and development occurred
and often with little knowledge or appreciation
that treaty conditions existed. As tribal ways
of life conflicted with non-native culture,
a number of laws were passed to minimize these
differences. These actions eroded tribal culture
and practices and by the 1960's a much larger
non-native population was openly clashing with
tribal fishing efforts. The regular arrest of
Indians, for carrying out what they believed
had been guaranteed to them in their treaties,
came to a head when the tribes turned to the
Federal court system to interpret their treaty
rights. This talk will briefly discuss the circumstances
leading up to this point, and the resulting
landmark court rulings on salmon and subsequently
on shellfish. You will hear this story, and
in particular the shellfish case and its consequences,
as seen by a non-native shellfish biologist
who has worked for 20 of those treaty tribes
on a reservation for almost 2 decades.
Born in Montreal, Quebec, Canada, David completed
a 2 year program at Dawson College, and then
went to McGill University for a Bachelor of
Science degree in Marine Biology. Undergraduate
research there focused on respiration in freshwater
mussels. In 1979 he moved to British Columbia,
to do his Masters degree at Simon Fraser University.
His research there dealt with geoduck clam recruitment
and required extensive SCUBA work, 30-60 feet
underwater, on the west coast of Vancouver Island.
After graduating, David undertook a number of
contract positions, including one at SFU, in
the Department of Mathematics and Statistics,
another in the University's Department of Biology,
a position with the International Salmon Commission,
and another with the environmental consulting
firm, Envirocon. In 1989, David and his wife
accepted a position with the Northwest Indian
Fisheries Commission, to set up a shellfish
program for the organization's member treaty
tribes. During that period he has also served
as Chairman of the Pacific Rim Shellfish Sanitation
Association, continues to serve on the Executive
Board of the Interstate Shellfish Sanitation
Conference, and has also served as the Chairman
of that organization's Foreign Relations Committee
for the last decade.
...and
our special keynote speakers:
Evolution
and Revolution in Analytical Science - from
the Bible to Alchemy to Single Molecule Detection
Gary D. Christian, Department of Chemistry,
University of Washington, Seattle, WA.
The teaching and practice of analytical chemistry
reflects the evolution of measurement science
over time. Qualitative and quantitative measurements
can be traced to pre-biblical times, and have
been important throughout the history of humans,
and today are key to the functioning of a modern
society. The perceived value of gold and silver
was the first incentive to acquire analytical
knowledge. The chemical balance is recorded
in the earliest documents found. I will trace
the development of analytical science, presenting
some of the pioneers through the eons, up to
those who formed the basis for many of our modern
techniques, and also early textbook authors
and how books evolved. Gravimetry emerged in
the 17th century, and titrimetry, along with
stoichiometric concepts, in the 18th and 19th
centuries. Quantitative analysis textbooks,
and hence the teaching of analytical chemistry
as a discipline, appeared in the 19th century.
The past century saw the development of instrumental
techniques, and we now possess incredible capabilities
for measurements.
Gary D. Christian received his B.S. degree in
1959 from the University of Oregon and Ph.D.
degree from the University of Maryland in 1964.
After serving as a research analytical chemist
at the Walter Reed Army Institute of Research
from 1961 to 1967, he joined the University
of Kentucky in 1967, and in 1972 moved to the
University of Washington as Professor of Chemistry.
He was Divisional Dean of Sciences in the College
of Arts and Sciences, 1993-2001. Christian's
research interests include electroanalytical
chemistry, atomic spectroscopy, and flow methods
of analysis. He is the author of over 300 papers
and has authored books on: Analytical Chemistry
(6 editions); Instrumental Analysis (2 editions);
Problem Solving in Analytical Chemistry; Quantitative
Calculations in Pharmaceutical Practice and
Research; Atomic Absorption Spectroscopy; and
Trace Analysis. Gary has been the recipient
of awards and other honors from several universities
and analytical societies in Europe and Asia.
His US honors include, among others, the ACS
Division of Analytical Chemistry Award for Excellence
in Teaching and the ACS Fisher Award in Analytical
Chemistry. He was Chairman of the Division of
Analytical Chemistry, 1989-90. He serves or
has served on the editorial boards of eighteen
journals and has been Editor-in-Chief of Talanta
since 1989.
Flow-Field
Flow Fractionation Applications from Biotechnology
to Nanotechnology
Pierluigi Reschiglian, University of Bologna,
Italy
Field-flow fractionation (FFF) is a family of
techniques able to separate macromolecules and
particles over a molar-mass range of 1014, with
particle size information obtained from retention
parameters. FFF has long been considered to
be the “best-kept secret” in the
field of separation science. This is because,
over more than 30 years, FFF has slowly evolved
from a research-oriented technique to a well-assessed
methodology. Today’s FFF is applied to
solve real analytical problems, particularly
when it is hyphenated to uncorrelated methods
for size, shape and/or mass characterization.
Because of the gentle mechanism, FFF separates
nanoparticles and complex bioanalytes in their
native state, which makes FFF particularly interesting
if further characterization needs to be performed
on the samples.
This presentation will provide an overview of
FFF basic principles and technology, and show
most recent applications to the analysis of
large molar-mass macromolecules and particles
of interest in nanotechnology and biotechnology.
From nanoparticles like carbon nanotubes, derivatised
and molecular imprinted nanobeads, nanoparticles
in the environment, and liposomes, from proteome
samples, prion aggregates, antibodies, and blood
lipoproteins, to viruses and even whole stem
cells: these are just some examples that indicate
nanotechnology and biotechnology as most suitable
application fields to finally make FFF a booming
methodology.
Dr. Pierluigi Reschiglian is a Professor of
Analytical Chemistry at the Department of Chemistry
“G. Ciamician”, Faculty of Science,
University of Bologna, Italy. He obtained his
BS and MS at the University of Ferrara, as well
as his PhD in Analytical and Environmental Chemistry.
His PhD work included 8 months at the Field
Flow Fractionation Research Center in Salt Lake
City, UT where he worked with Prof. J.C. Giddings,
inventor of Field Flow Fractionation (many familiar
with Giddings as distinguished chromatographer
and theorist)
Although his research interests are varied,
including many aspects of separation science,
light scattering and mass detection among other
detection methods and computational aspects,
Dr Reschiglian is now best known for his work
with Field Flow Fractionation methods for proteins
and nanoparticles. The research is both fundamental
and applied in nature and ranges from studies
of proteins expressed by microbes to studies
of nanotubes and other nanoparticles. He has
functioned as Director in many international
collaborative projects, is the author of more
than 100 papers, and has also invented novel
methods for separating eukaryotic and prokaryotic
cells.
Tales
From The Trenches -- The Spinach Outbreak 2006
Linda Guthertz, California Department of
Public Health
During September 2006, a serious foodborne outbreak
caused by E. coli O157:H7 resulted in 204 cases
including 3 deaths. This outbreak was traced
back to spinach product that had been grown,
harvested, and processed in the San Juan Valley
of California. The traceback investigation further
narrowed the possible sites of contamination
to four fields on four ranches. The scope of
this investigation required the use of the California
Food Emergency Response Team (CalFERT) which
is a co-operative group made up of scientists
and investigators from the Food and Drug Administration
(FDA) and the California Department of Public
Health (CDPH). Scientists from the USDA-Agricultural
Research Service (ARS) also participated. Laboratories
from each of these institutions were called
upon to share the analyses of the 879 samples
collected by investigators from field examinations.
Using a newly validated Recirculating Immuno-magnetic
Separation method (RIMS), the CDPH-Food and
Drug Laboratory Branch (FDLB) analyzed 23.9%
of the total number of samples collected. Of
these specimens, 36.7% were water, 17.6% were
environmental swabs, 10% were soil/sediment,
8.6% were field products, and 27.1% were animal
feces. Using RIMS methodology, E. coli O157:H7
was recovered from 1.3% of water samples, 7.9%
of swabs, and 35.1% of animal droppings. Among
stx2 E. coli O157:H7 isolates analyzed by pulsed
field gel electrophoresis (PFGE) and/or multiple
locus variable number tandem repeats analysis
(MLVA) by the Microbial Diseases Laboratory
(MDL), 26.2% of the isolates (from 7 positive
samples) matched the genetic pattern of the
outbreak strain. The RIMS procedure can be found
on the FDLB website at: http://www.dhs.ca.gov/fdlb/microbiology.
Paralytic
Shellfish Poisoning Toxin Detection in Human
Urine and Serum: 2007 Outbreak
Stacey Etheridge and Mark Poli
Toxin detection methods are typically validated
for extracts from the causative algae and/or
the affected seafood. There are presently no
officially validated methods for detecting marine
toxins in clinical matrices. In this work, the
AOAC Lawrence HPLC method for determining paralytic
shellfish poisoning (PSP) toxins in shellfish
was evaluated for the human clinical matrices
urine and serum. Initial analysis revealed an
interfering, naturally-occurring fluorescent
compound in urine. Further analysis by high
resolution mass spectrometry identified the
compound to be hippuric acid, a major constituent
in human urine originating from dietary sources.
The hippuric acid was removed from samples by
adjusting the pH to 4 prior to sample clean-up
and by doubling the SPE cartridge bed volume.
Interference by naturally-occurring fluorescent
compounds was found to be minimal in the serum
matrix. Quantitation of a range of PSP congeners
spiked in these matrices was determined and
will be presented. This method was later applied
during an actual PSP outbreak in 2007 to analyze
human urine and serum from four patients after
they became ill following consumption of toxic
mussels from a floating barrel off the coast
of Maine. The Lawrence HPLC method, with minimal
modifications, was used to identify and quantify
toxin composition and concentrations. Urine
was found to be the matrix of choice for detecting
PSP congeners and elimination rates of the main
toxins were determined. Results indicate that
this method could be employed during future
PSP outbreaks to detect toxins in clinical samples,
which would further lead to an improved knowledge
base about human health effects from intoxication.
Stacey Etheridge earned a Bachelor of Science
in Biology from the University of Alabama in
1994. She also received a Master of Science
and a Doctorate in Oceanography from the University
of Connecticut in 1997 and 2002, respectively.
Stacey spent a year as a post-doctoral staff
fellow at the FDA’s Center for Food Safety
and Applied Nutrition (CFSAN) where she conducted
research on marine biotoxin detection methods.
She is currently a Biologist in the Spectroscopy
and Mass Spectrometry Branch of the Office of
Regulatory Science at CFSAN. Stacey performs
research related to food safety and food defense,
including developing detection methods for foodborne
toxins, investigating traditional and emerging
sources and vectors of foodborne toxins, and
studying the dynamics of toxic algae and the
transfer of toxins to seafood. Stacey has authored/co-authored
numerous publications on toxin detection, harmful/toxic
algae, and bio-optical detection of algal blooms.
She has also co-edited a book on harmful algae
management and mitigation and has co-authored
reports to Congress on the scientific assessment
of harmful algal blooms and oceans and human
health. Among her many research projects, she
currently oversees FDA’s contribution
in a large, multi-laboratory interdisciplinary
investigation on toxic algal blooms and shellfish
toxicity (associated with paralytic shellfish
poisoning) off the coast of New England, the
goal of which is to improve management of this
seafood safety hazard. Stacey collaborates with
other federal agencies such as USAMRIID and
NOAA, academic and non-profit organizations,
and industry. She is a member of the AOAC’s
Presidential Task Force on Marine and Freshwater
Toxins, an FDA representative on the Interagency
Working Group on Harmful Algal Blooms, Hypoxia
and Human Health (IWG-4H), and an FDA Team Member
on the ISSC (Interstate Shellfish Sanitation
Conference) Biotoxin Committee.
Dr. Poli earned a Master of Science Degree in
Oceanography from the University of Miami Rosenstiel
School of Marine and Atmospheric Sciences in
1982, followed by a Doctorate in Biochemistry
from the University Of Miami School Of Medicine
in 1986. He spent three years as a National
Research Council postdoctoral fellow at the
US Army Medical Research Institute of Infectious
Diseases at Fort Detrick, in Frederick, Maryland
where, in 1988, he accepted a full-time position
as Principal Investigator in what is now the
Integrated Toxicology Division. Dr. Poli received
his Board certification in General Toxicology
from the American Board of Toxicology in 1990
and is currently adjunct faculty at Hood College
in Frederick where he teaches Environmental
Toxicology in the Environmental Science graduate
program. For the past 20 years his research
has centered on mechanism of action and detection
of a variety of natural toxins. He has authored
numerous research articles in scientific journals
as well as review chapters in several books,
including the Encyclopedia of Bioterrorism Defense
and the Textbook of Military Medicine. He lectures
routinely to military and civilian audiences
on the subject of toxins as weapons of mass
destruction, and serves as the USAMRIID subject
matter expert on toxins to law enforcement groups
and other government agencies. He also collaborates
extensively with the US Food and Drug Administration
and the National Ocean Service in the development
of detection methodologies for marine toxins,
is a member of the AOAC’s Presidential
Task Force on Marine and Freshwater Toxins,
and in 2006 was appointed the AOAC’s General
Referee for Biological Toxins.
Critical
Aspects for Successful ELISA Development
Lyn R. Briggs, Toxinology, AgResearch, Ruakura
Research Centre, New Zealand
The use of ELISAs for research and monitoring
tools is widespread in the control of natural
and industrial contaminants and it is now possible
to purchase test kits for a wide range of these
compounds. In the past, emphasis was on the
application of fully quantitative immunoassays
for research, and the methodology proved to
be invaluable particularly in projects requiring
the analysis of large numbers of samples and
also in situations where analyte levels were
lower than the detection limits of existing
methodologies. Development of immunoassays is
primarily dependent on the ability to produce
antibodies that give the sensitivity and specificity
required for an assay. Both of these requirements
can only be met by major inputs in toxicology
and chemistry – particularly for the isolation
of milligram quantities of the analyte of interest
for preparation of immunoreagents and later
highly purified material for use as calibration
standards in the assays. Recently selected research
ELISAs were reformatted to develop test kits
or to transfer into commercial analytical laboratories.
An outline of the practicalities involved in
immunisation programs, antibody screening, assay
optimisation and in-house application will be
given to illustrate some of the aspects of successful
ELISA development
Dr. Lyn Briggs is Senior Scientist in Toxinology
at the Applied Biotechnologies Group of AgResearch
of New Zealand. She holds BSc degrees in both
Zoology and Chemistry from Auckland University,
and at Waikato University, the MPhil, and PhD
degrees (Biological Sciences, Immunochemistry).
At AgResearch she conducts algal and endophyte
fungal toxin research applying immunotechnologies.
Her work includes monoclonal and polyclonal
antibody production and, ultimately, development
and validation of extraction procedures and
immunoassays. The assays are developed to quantify
toxins in algae, shellfish, water, culture supernatants,
plant material, animal fluids and tissues. Lyn
has served as project manager in monitoring
programs for cyanobacteria and on other, international
collaborative projects with the US and Norway,
with some of the resulting immunochemical methods
becoming commercialized ELISAs. Other methods
have been used to establish quality control
programs for novel endophyte pastures. Lyn also
serves as Topic Advisor on Immunochemical Methods
with the AOAC Marine and Freshwater Toxins Task
Force, and has authored numerous publications.
The
NIH/ODS Analytical Methods and Reference Materials
Program for Dietary Supplements: Accomplishments
and Future Directions.
Joseph M. Betz, Office of Dietary
Supplements, National Institutes of Health,
Bethesda, MD, 20892 USA
Quality of botanical products is one of the
greatest uncertainties that consumers, clinicians,
regulators, and researchers face. Definitions
of quality abound, and include specifications
for sanitation, adventitious agents (pesticides,
metals, weeds), and content of natural chemicals.
Validated analytical methods and reference materials
to ensure the identity, purity, quality, and
strength of constituents in natural health products
and dietary supplements are essential. Researchers
need these materials and methods to characterize
test articles used in research and ensure that
the materials are of sufficient quality that
studies can be reproduced. Regulators and industry
need them in dealing with regulatory, safety,
labeling, quality control, and manufacturing
issues. Because these products and their ingredients
are often complex mixtures, they pose analytical
challenges and methods validation may be difficult.
In response to widespread concerns about product
quality and the need for validated and publicly
available methods for DS analysis, in 2002 the
U.S. Congress directed the Office of Dietary
Supplements (ODS) at the National Institutes
of Health (NIH) to accelerate an ongoing methods
validation process, and the Dietary Supplements
Methods and Reference Materials Program was
created. The program was constructed from stakeholder
input and incorporates several U.S. federal
procurement and granting mechanisms in a coordinated
and interlocking framework. The framework facilitates
validation of analytical methods, analytical
standards, and reference materials. The major
accomplishments of the first five years of the
Dietary Supplements Methods and Reference Materials
program are collaborative efforts with FDA,
AOAC, and NIST. The ODS/FDA/AOAC project has
resulted in 18 collaborative studies of methods
for dietary supplement constituents. Seven of
the studied methods have been approved as First
Action Official Methods of Analysis (OMA) and
3 additional methods have been approved as Final
Action OMA. An additional 6 collaborative study
reports were being reviewed by the AOAC Official
Methods Board as of 3/31/08. The ODS/FDA/NIST
project has resulted in the production of 5
suites of dietary supplement Standard Reference
Materials, with an additional 12 suites in various
stages of completion. NIST has also created
a pilot Dietary Supplement Laboratory Quality
Assurance Program that will assist participating
laboratories to become proficient at dietary
supplement analysis. A more detailed account
of these accomplishments and an outline of the
future scope and direction of the program will
be presented.
Joseph M. Betz, Ph.D. joined the ODS as Director
of the Dietary Supplements Methods and Reference
Materials Program in 2001. In this role, he
oversees efforts to promote development of validated
analytical methods and reference materials for
dietary supplements. Prior to joining NIH, he
was Vice President for Scientific and Technical
Affairs at the American Herbal Products Association.
Before joining the AHPA, he spent 12 years as
a research chemist at FDA’s Center for
Food Safety and Applied Nutrition. Dr. Betz
is a native of Philadelphia, Pennsylvania. He
earned his Ph.D. in Pharmacognosy (1988) at
the Philadelphia College of Pharmacy and Science.
His research interests lie in the areas of analytical
methods for determination of botanical quality.
He has served as AOACI General Referee for Botanical
Supplements and on two USP Expert Committees-
Botanicals and Nomenclature and Labeling. Dr.
Betz has over 50 publications in the natural
products area and continues to work toward ensuring
the quality and safety of botanical products.
Dr. Betz is the recipient of the American Botanical
Council’s first Norman R. Farnsworth award
for excellence in Botanical Research and the
American Herbal Product Association’s
Herbal Insight Award for contributions to the
Botanical Sciences.
Micro-Total
Analytical Systems and other Field-capable Platforms:
Challenges and Considerations
Victoria VanderNoot, Sandia National
Laboratories, Livermore,CA
Sandia National Laboratories in California has
been developing automated biological point detection
systems for more than a decade, building on
its strong engineering base. The ?ChemLab™,
incorporated two chip-based electrophoresis
laser induced fluorescence (CE/LIF) units into
a single hand-held, battery powered device.
It was originally developed with first responders
in mind and analyzed fluorescently-labeled proteins,
or other molecules, in five minutes. Since the
initial effort, we have built upon the original
?ChemLab™ technology to adapt it to a
variety of sensing applications and have developed
of a series of fieldable detection platforms.
Among these are the Automated Microfluidic Protein
Profiling System (AMPPS) developed to detect
and identify aerosolized organisms via their
protein signatures; the Unattended Water Sensor
(UWS), developed in collaboration with Tenix
Corporation for monitoring domestic water supplies;
and BioBriefcase (BBC), an automated immunoassay
and PCR analysis system developed in collaboration
with researchers at Lawrence Livermore Laboratories.
All of these systems incorporated automated
biological sample preparation and analysis for
fieldable, stand alone detection platforms.
We will discuss our efforts towards bringing
analytical instrumentation to the field, highlighting
a few of our platforms in the context of homeland
security considerations, systems analysis, systems
engineering and concepts of operation (CONOPS).
Victoria VanderNoot received both a B. Sc. (Chemistry)
and a Ph. D. (Analytical Chemistry) from Carleton
University in Canada where her thesis work focused
on optical biosensor development employing photothermal
spectrometry-based detection. After several
years postdoctoral research experience in the
areas of bioassay development and electrophoresis
at University of Colorado Health Sciences Center
and then University of Iowa's Department of
Chemical and Biochemical Engineering, she joined
Sandia National Labs where she is currently
a Principal Member of Technical Staff in the
Biosystems Research Department. Her research
interests focus on microfluidics, and sensing
technologies for toxins, proteins and other
biomolecules. For the last 10 years she has
been part of a highly interdisciplinary group
of scientists and engineers working to develop
integrated and fieldable devices for environmental,
biomedical, and biodefense applications.
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...and
a word from our AOAC sponsors:
(TRAINING)
Single Laboratory Validation of Microbiological
Methods
Sharon L. Brunelle, Ph.D., Brunelle Biotech
Consulting, Technical Consultant to AOAC Research
Institute
------------------------
Sharon
L. Brunelle, Ph.D., is the owner of Brunelle
Biotech Consulting in Woodinville, WA. She consults
mainly in the area of food safety diagnostics.
Dr. Brunelle is an ongoing technical consultant
for the AOAC Research Institute and AOAC INTERNATIONAL.
Through AOAC, she has provided technical expertise
for AOAC contracts with the US Department of
Homeland Security for the validation of methods
for Bacillus anthracis and validation of an
ASTM-AOAC Sampling Standard for Visible White
Powders; is AOAC’s lead microbiologist
for their contract with the US FDA for Best
Practices in Microbiological Methods Validation;
and is project coordinator for the Performance
Tested MethodsSM and Official Methods of AnalysisSM
programs.
Prior to consulting, Dr. Brunelle was Chief
Scientific Officer at Molecular Circuitry in
King of Prussia, PA. Molecular Circuitry owns
patented biosensor technology, which was applied
to the detection of pathogens in food and environmental
samples. Before joining Molecular Circuitry,
Sharon worked in the clinical diagnostic field,
first as a scientific manager at Bard Diagnostic
Sciences, at the time a division of C.R. Bard
located in Redmond, WA, then as an independent
consultant for various clinical diagnostic biotech
companies. Her specialty was clinical oncology
and while at Bard, she developed and received
FDA 510(k) approval for a new rapid immunoassay
for bladder cancer. Prior to Bard, Dr. Brunelle
was a lead scientist at BioControl Systems,
now in Bellevue, WA, where she developed new
immunoassays for food pathogens.
Dr. Brunelle received her B.S. in chemistry
and biology at the University of Delaware and
her Ph.D. in biochemistry at Brandeis University.
Her thesis work was in the area of enzyme mechanisms
and rational drug design. Dr. Brunelle’s
postdoctoral training was carried out jointly
at Rockefeller University and the Picower Institute
of Medical Research in medical biochemistry,
investigating the biochemical causes of diabetic
complications.
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