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Friday, August 20, 2010
Marine Environmental Research Institute GULF ECOTOX (GET) PROJECT: Protocols for Sample Collection
THANKS TO MICHELLE NIX AND GULF COAST OIL SPILL VOLUNTEERS!
Marine Environmental Research Institute
GULF ECOTOX (GET) PROJECT: Protocols for Sample Collection
The Marine Environmental Research Institute (MERI) and its partners are proposing a collaborative, regionwide
investigation of toxic impacts of oil and dispersants throughout the Gulf ecosystem - from water,
sediments, and phytoplankton to fish and marine mammals. This broad-based, multi-species, multi-habitat
approach can provide essential information that will complement existing efforts to measure ecosystem
damage, inform public health (seafood safety), and guide future restoration.
Sampling Approach
The following is a preliminary program designed to guide the collection of samples in diverse media for the
analysis of toxic contaminants related to oil and dispersants. As we continue assembling our team of partners
and developing the research plan, sampling protocols will be expanded and refined. This approach is intended
to complement other monitoring efforts in the Gulf. We will build on available data from existing federal and
state monitoring programs and avoid duplication of effort wherever possible. As appropriate, the project can
complement analyses by in situ regional laboratories and provide external confirmation of results.
Sample collection needs to begin as soon as possible to capture impacts during this high exposure
period. We are enlisting the cooperation of state and federal agencies, stranding networks, and trained
volunteers to obtain environmental and biotic samples across the Gulf. Sampling kits with detailed sampling
instructions will be distributed region-wide through the Gulf Restoration Network and key agencies and
universities. Volunteer training will be offered at selected locations, and training videos will be made available.
Water Sampling 1: Analysis of Water Containing Dispersant and Oil
An immediate objective is to obtain samples of water containing Corexit dispersant for comprehensive
analysis of the specific ingredients. Not all the chemical ingredients in Corexit have been identified and their
breakdown and interaction by-products in seawater are unknown.
Water samples will be collected from several spatial locations to represent hot spots, moderately
contaminated zones and reference areas for comparison.
1. Collect 10 grab samples of water (50 – 100 ml each) in polyethylene bottles from a location where
dispersant is concentrated (recently applied) in the water.
2. Collect five grab samples of water (50 – 100 ml) from three areas where dispersant appears to be
moderately concentrated (total of 15 samples).
3. Collect 5 grab samples of water (50 – 100 ml) from three relatively clean sites (total of 15 samples) in the
same general area.
Fill polyethylene bottles approximately ¾ full; do not fill to the top. Record date and time and lat/longs for
each water sample collected. Provide description of location, noting approximate distance from the discharge
source.
Freeze samples at – 20 degrees (regular freezer) as soon as possible and kept frozen until all samples can be
shipped overnight to the lab. While on the boat, keep the samples cool on ice.
Ship frozen samples to the analytical laboratory on dry ice by overnight carrier (Federal Express). Bottles can
be placed inside a foam shipping container wrapped in bubble wrap. DO NOT SHIP OVER A WEEKEND.
Use plenty of dry ice so the samples stay frozen until they reach the lab (see shipping instructions below).
Water Sampling 2: Water and Sediments*
*Preliminary protocol to be expanded in collaboration with federal agencies and university research groups
We plan to collect water and sediment from the same locations concurrently for studying partitioning of
chemicals between water and sediment and to evaluate if sediment acts as a reservoir for some of the
toxicants that would result in long term health effects on marine biota.
We propose to scan different types of contaminants that can be present in water. Because samples are collect
along a contaminant gradient, we will be able to identify those contaminants that are released from Corexit
and from oil and the levels of those components. PAHs including alkylated PAHs will be determined as
target analytes. We will also identify other contaminants that are released from oil and dispersant application.
Water samples will be collected from several spatial locations to represent hot spots, moderately
contaminated zones and reference areas for comparison.
1. Collect 10 grab samples of water in polyethylene bottles (100 ml to 1 L) from locations that represent hot
spots of oil contamination.
2. Collect 5 grab samples of water (100 ml to 1 L) from 3 moderately contaminated areas (total of 15 samples).
3. Collect 5 grab samples of water (100 ml to 1 Ll) from 3 reference areas (total of 15 samples).
Fill polyethylene bottles approx. ¾ full; do not fill to the top.
Record date and time and lat/longs for each water sample collected. Provide description of location, noting
distance from the discharge source. Freeze water samples and store at – 20 degrees (regular freezer) as soon
as possible until shipment overnight to the analytical laboratory (see shipping instructions below).
Sediment Sampling*
*Preliminary protocol to be expanded in collaboration with federal agencies and university research groups
Sediment samples will be collected from the same location where water samples were collected (above). In
addition, we propose to collect sediment samples from several marshlands/wetlands which are habitats for
various wildlife species. We expect to delineate spatial distribution of contaminants including PAHs and
alkylated PAHs in sediments and the magnitude of contamination in coastal wetlands.
Collect surficial sediment samples from several spatial locations to represent hot spots, moderately
contaminated zones and reference areas for comparison. Subsurface sediments (0-1”) including oil will be
collected using a ponar grab sampler. Subtidal sediments will be sampled at the same locations where water
samples were collected (see above).
Record date and time and lat/longs for each sediment sample collected. Provide description of location,
noting distance from the discharge source. Freeze sediment samples and store at -20! until shipment to the
analytical laboratory (see shipping instructions below).
Plankton*
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*Sampling protocol to be expanded in collaboration with federal agencies and university researchers
Phytoplankton and zooplankton samples will be collected from several spatial locations will be collected to
represent hot spots, moderately contaminated zones and reference areas for comparison.
Surface samples of seawater will be collected and drained through a 20 micrometer PVC sieve. Collect a total
of 10-20 samples per location for the pilot study. Send 10 ml of each sample to the lab for FlowCAM imaging
and quantitation of phytoplankton and zooplankton particles.
Freeze plankton samples (10-20 ml) for contaminant analysis and store at -20! until shipment to the analytical
laboratory for analysis (see shipping instructions below).
Shellfish
Bivalves and crustaceans (oysters, crabs, shrimps, spiny lobster) will be sampled in collaboration with
appropriate state and federal fisheries agencies. Samples from several spatial locations will be collected to
represent hot spots, moderately contaminated zones and reference areas for comparison. These samples will
be pooled, homogenized, and analyzed for the presence of PAHs and OH-PAHs.
For bivalves, 20 individuals should be hand-picked at each site (size range to be determined). Samples from a
given location will be pooled to obtain a representative sample for that location. Record shell length, and keep
in a cooler box on ice until samples can be frozen.
For small crustaceans, collect 5-10 individuals at each site. For spiny lobster, collect 1-2 individuals per site.
Wrap in aluminum foil, shiny side out. Record shell length, and keep samples in a cooler box on ice until they
can be frozen.
Record date and time and lat/longs for each batch of samples collected. Batch labels should show date and
time collected, location in lat/long, and species. Provide description of location, noting distance from the
discharge source. Freeze samples and store at -20! until shipment to the analytical laboratory (see shipping
instructions below).
Forage Fish – Estuarine and Marine
Fish (black drum, red drum, sea trout, sheepshead, red herring, menhaden) will be collected in collaboration
with appropriate state and federal fisheries agencies to determine the short-term and long-term impacts of oil
and dispersants. Fish from several spatial locations will be collected to represent hot spots, moderately
contaminated zones and reference areas for comparison. These samples will be pooled, homogenized, and
analyzed for the presence of PAHs and OH-PAHs.
Collect 10 small fish (whole fish) per species per site sampled. Wrap whole fish in foil (shiny side out), place
batches of 10 fish (same species) in labeled Ziplock bags, keep in a cooler box on ice and freeze as soon as
possible.
Labels should show date and time collected, location in lat/long and species, number of fish in batch, and
approx. length and weight. Provide description of location, noting distance from the discharge source..
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Store frozen fish in freezer at -20 degrees until several batches are ready for shipment overnight to the
analytical laboratory (see shipping instructions below).
Marine Fish
From large marine fishes (cobia, amberjack, red snapper, mangrove snapper, barracuda, yellowfin tuna), fillet
(muscle) and liver tissues will be collected for the analysis of PAHs and OH-PAHs. Fish from several spatial
locations will be collected to represent hot spots, moderately contaminated zones and reference areas for
comparison.
Collect 1-5 large individual fish per species per site. At necropsy using a sterile knife, cut open the fish body
and collect the liver (whole liver or 100 g), and 100 g of muscle with skin intact. Wrap liver and muscle tissue
from each individual fish separately in aluminum foil (shiny side out), place tissues in a labeled Ziplock bag,
keep samples on ice in the field and freeze as soon as possible.
Labels should show date and time collected, location in lat/long, species, and specific tissue. Note whether
collection site was oiled or relatively clean.
Store frozen fish in freezer at -20 degrees until several batches are ready for shipment overnight to the
analytical laboratory (see shipping instructions below).
Sea Turtles
Sea turtles in the Gulf are endangered or threatened and are managed under NOAA/NMFS. We will
collaborate with this agency and local stranding personnel to obtain samples for the analysis of PAHs and
OH-PAHs in tissues of dead turtles, as well as dispersant ingredients which may impact the survival of these
vulnerable species.
Do NOT samples dead sea turtles in the field. Transport dead sea turtles to the nearest rehabilitation center
(Audubon Aquarium) for necropsy and sampling. From fresh dead sea turtles, we request liver samples (100
ml) for analysis of PAHs and OH-PAHs. Store liver tissue in labeled I-Chem jars and freeze as soon as
possible.
Labels should show date collected, location in lat/long, species, and tissue. Note condition of the animal and
whether collection site was oiled or relatively clean.
Freeze tissues and store at -20 degrees until several batches are ready for shipment overnight to the analytical
laboratory (see shipping instructions below).
Sea Birds
Marine birds and shorebirds (pelicans, terns, gulls) are managed by US Fish & Wildlife Service. We will enlist
cooperation from agency personnel to collect these samples.
From stranded dead birds, collect liver samples (100 ml) and a feather for toxicological studies. Store liver
tissue in labeled I-Chem jars and freeze as soon as possible. Wrap feather in aluminum foil (shiny side out),
place in labeled Ziplock bag, and freeze.
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Labels should show date and time collected, location in lat/long, species, and tissue. Note condition of the
animal and whether collection site was oiled or moderately, or relatively clean.
Freeze tissues and store at -20 degrees until several batches are ready for shipment overnight to the analytical
laboratory (see shipping instructions below).
Marine Mammals
Marine mammals such as bottlenose dolphins and sperm whales are protected species managed by NOAA
NFMS. West Indian manatees are a highly endangered species. We will enlist cooperation of regional NMFS
stranding networks to obtain tissue samples from marine mammals that strand or die in the Gulf.
Animals must be fresh dead (Code 2 or 3).From stranded fresh dead marine mammals, collect blubber, liver,
kidney, and muscle samples (50-100 ml each) for analysis of PAHs and OH-PAHs, other relevant compounds.
Store tissues in labeled I-Chem jars and freeze as soon as possible.
NMFS Level A data is requested on each individual animal. Labels should show date and time collected,
location in lat/long, species, and tissue. Note condition of the animal and whether collection site was oiled or
relatively clean.
Freeze tissues and store at -20 degrees until several batches are ready for shipment overnight to the analytical
laboratory (see shipping instructions below).
Shipping Instructions
Ship all frozen samples to the laboratory on dry ice via overnight carrier (Federal Express). Samples can be
placed in a foam shipping container or wrapped in bubble wrap. Use plenty of dry ice so the samples stay
frozen.
Do NOT ship on Friday or over a weekend. If samples are collected on Thursday or Friday, keep
frozen until Monday and ship overnight. *Make sure to call Dr. Kannan or his project manager, Dr. Qian Wu
(wuqiansjtu@gmail.com) and let him know samples are on the way.
Ship samples overnight to:
Wadsworth Center, NY State Dept of Health
ATT: Dr. Kurunthachalam Kannan or Dr. Qian Wu
Chief, Laboratory of Organic Analytical Chemistry
Empire State Plaza, Dock J – P1 Level, Albany, NY 12237
Tel: 518-474-0015
For information, contact:
Susan D. Shaw, DrPH
Director, Marine Environmental Research Institute (MERI)
Center for Marine Studies, PO Box 1653, 55 Main Street
Blue Hill, ME 04614 USA
Tel: (207) 374-2135 Cell (212) 203-9539 Fax: (207) 374-2931
Email sshaw@meriresearch.org
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http://www.meriresearch.org
Kurunthachalam Kannan, Ph.D.
Chief, Laboratory of Organic Analytical Chemistry
Wadsworth Center, NY State Dept of Health
Empire State Plaza, PO Box 509
Albany, NY 12201-0509, USA
Tel: 518-474-0015; Fax: 518-473-2895
Email kkannan@wadsworth.org
http://www.wadsworth.org/resnres/bios/kannan.htm
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