SUBMISSION FORM for Tissue Collection
Environmental Conservation Division, Northwest Fisheries Science Center, NMFS/NOAA, 2725 Montlake Blvd East, Seattle, WA 98112
Background Information:
Organization submitting samples:_____________________________, Individual collecting samples:________________
Animal identification/tag number and/or name:____________________________________________________________
Species: Scientific name:
Common name: ________________
_
Dates of Stranding:________________, of Death:_________________, of Tissue collection:_______________________
Location of stranding:________________________________________________________________________________
Name or location of the pod/colony the animal was from (if known or applicable):________________________________
Age (in years, if known, or use neonate, pup/calf, juvenile, adult):____________________________________________
Size: Length (snout to tail tip/fluke notch)__________, Weight (estimate)___________ or (actual)__________________
Axillary girth (half girth if large cetacean)_________
Sex (if adult female please note reproductive status): Male____, Female____, Pregnant?____, Lactating?____, Neither___
Body condition (please check one): Emaciated_____, Fair_____, Good_____, Robust_______
Blubber thickness (measure over the xyphoid cartilage, excluding the skin)_______, Oil leaching from blubber?___
Feeding status: Entire digestive tract empty______, Stomach empty but intestines contain some material_______,
Recent meal (please list type and amounts of stomach contents if ingested in the wild)________________________
Cause of death (if known):
Tissue Preservation: Wrap each tissue sample in a rinsed Teflon sheet and then place each bag in a separate
labeled Whirl-pak® bag. Seal and place on ice. Freeze as soon as possible.
Tissues Requested: Please check which samples were taken and indicate the location taken from.
_____1. Blubber - Full thickness (from skin to muscle) taken from over the xyphoid cartilage, if possible, 100-200 grams.
Location, if different than indicated:
2. Liver - 100 grams. Location sample taken from:
_____3. Kidney - 100 grams. Location sample taken from:
Additional History (including behavioral observations, treatment records, and necropsy results, if
available, any abnormalities found, and any other information available about the animal). Please use the
back of this page and/or attach additional sheets and records. Thank You.
February 12, 2016
Marine Mammal Tissue Sampling Protocol
Tissue Sampling for Chemical Contaminant Analyses
Supplies for sampling will include:
— 12" x 13" acetone rinsed Teflon sheets
— 18 oz. Whirl-pak® bags (4.5" x 8.5")
— "Rite-in-the-Rain" tags for inside Whirl-pak® bags
— rubber bands
— ballpoint and marking pens
— submission forms on "Rite-in-the-Rain" paper
Sampling Protocol
• Priority for collection of samples is: blubber, liver, kidney (see attached Marine Mammal Sampling Sheet/Data
Record for description of how and from where to collect tissue samples).
• See attached Marine Mammal Tissue Sampling Protocol Addendum for additional information on sampling
procedure.
• For tissue collection, use a stainless steel knife and rinse the knife between necropsies of each animal.
• Wrap individual tissue samples in Teflon sheet, secure with rubber band, and then put into
labeled Whirl-pak®
bags. Label the bags with:
Animal ID Number___________________
Species____________________________
Tissue Type________________________
Date Collected______________________
• Insert labeled (as above) "Rite-in-the-Rain" tags into each Whirl-pak® bag (this is in case the writing comes off the
Whirl-pak® bag). Remove as much of the air as possible from the bag before it is sealed. Place samples on ice. As
soon as possible, freeze at lowest
temperature available.
• Please provide copy of full necropsy report (see attached Marine Mammal Sampling Sheet/Data
Record).
Shipment of Samples
• Ship frozen samples, early in the week, with ~5 lbs dry ice, FedEx overnight to: Karen Tilbury, NWFSC, ECD, 2725
Montlake Blvd. E., Seattle, WA 98112-2097. Call Karen (206-860-3338) the day the samples are shipped with
the invoice number for tracking, if necessary.
February 12, 2016
Marine Mammal Tissue Contaminant Analyses
Environmental Conservation Division, Northwest Fisheries Science Center,
National Marine Fisheries Service, National Oceanic and Atmospheric Administration,
2725 Montlake Blvd. East, Seattle, WA 98112-2097
Phone: 206-860-3330 FAX: 206-860-3335
MARINE MAMMAL TISSUE SAMPLING PROTOCOL
Tissue collection procedures - It is important to use standardized sampling procedures so that, even when there
are low levels of contaminants present, the differences may be attributed to biological processes and
contaminant exposure and not to variation in the collection process. The following procedures are essential to
prevent cross-contamination of tissues within an animal and ensure uniformity of samples among animals.
1. Collect tissues only from animals that have been dead less than 24 hours. Even within 24 hours
after death, the autolytic changes may become severe enough to confound efforts to interpret the
contaminant data. This is why accurate information about the time between death and tissue
collection is important.
2. Collect liver, blubber, and kidney samples from the locations specified on the submission form,
if possible. This reduces variation caused by possible composition differences within tissues of
the same animal. It also provides us with uniform samples and information from all participating
organizations which can be directly compared based on the demographics of the animals.
Sample size: blubber; 100 - 200 g (10 g minimum which is the full thickness of blubber and
about the area of a credit card), liver; 100 g (size of a small human fist), kidney; 100 g (size of a
small human fist).
3. Rinse all instruments with isopropyl alcohol before each tissue is sampled. This will minimize
cross-contamination of tissues. Particular care needs to be taken after cutting through the skin
(because of possible environmental contamination) and blubber (because of the high lipid
content). When collecting blubber, remove the skin to prevent contamination, but include the
full thickness of blubber from skin to muscle. Ideally, separate instruments would be used and
scalpel blades changed after skin and blubber are cut.
4. Treat sample containers provided as if they are sterile. The teflon sheets have been rinsed with
acetone, and then dried in a dust free environment before shipment to your facility. This is to
reduce the possibility of external contamination before and during the sampling procedure by any
of the compounds we are studying.
5. Keep samples as cold as possible after collection. Some of the organic contaminants are volatile or
are degraded by compounds released during cell death, particularly in the liver and kidney. To
decrease changes in contaminant levels due to these processes, keep the samples on ice following
the necropsy and place in a freezer as soon as possible.
February 12, 2016