Genetic Tissue Bank Sampling Protocol

15th Jan 2008 The following guide is designed to instruct you on how to safely remove a skin sample from a dead stranded cetacean, package it, label it and return it to the Natural History Museum in Dublin for storage in the National Cetacean Genetic Tissue Bank.

Please remember that all dead animals represent a health risk.

Gloves (provided in kit) should be used at all times and you should wash your hands after obtaining the sample.

Kit Contents:



1 Plastic kit container with screw top lid (issued with first kit only)

1 Instruction sheet

10 Data record cards & 10 Stranding Record Forms

10 Sample tubes containing 99% Ethanol (clear liquid- DO NOT DRINK)

10 Pairs of Nitrile safety gloves

10 Resealable plastic bags (when returned, each should contain 1 sample tube and 1 record card per individual animal)

10 Disposable scalpels

10 Padded addressed envelopes (postage paid)

Do not store your kit near a heat source (direct sunlight, radiator, warm fan). Preferably store the kit in a cool dry location.

Instructions:

1. Before touching the animal, don a pair of Nitrile gloves!

NOTE: Please use a fresh pair of gloves and a new sterile scalpel (and subsequent sample tube, data card, bag) for each individual animal to prevent cross-contamination of genetic material from different animals.

2. Use a scalpel (knife) provided in the kit to slice skin (usually a greyish colour, please do not sample the blubber which is a yellow-whitish colour) from the body of the animal. The skin only (top few millimetres) is required - no blubber should be attached.

Two skin strips approx. 1cm X 2cm (small enough to fit into the sample tube, NOT filling all of the way) should be taken.


An example of blubber tissue (yellowish colour). This should not be included in samples for the tissue bank.

3. Carefully unscrew the top of the sample tube containing the 99% Ethanol and place the tissue sample in the tube. PLEASE NOTE: only samples from one animal are to be placed in each tube and only one record card is to be used for each animal sampled.

Do not overfill the sample tube thereby displacing all the ethanol. Take a skin sample preferably without the blubber layer - if skin is unavailable, please try to retrieve some muscle tissue (rich dark red colour) that is roughly the size of your thumbnail and cut into two pieces.

Skin sample© Ruth Carden

A skin sample (greyish colour) returned to the Museum, along with record card. There are three strips of tissue which occupy more than half of the 2ml sample tube. This tube is overfilled with tissue and half of this sample would be sufficient for the tissue bank.


4. Carefully replace the lid on the sample tube and screw tight. The lid is fitted with a rubber O-ring, which prevents evaporation of the ethanol.

5. Using the pencil, please fill out the enclosed data record card with the following data:

· Date of sampling & Date of stranding (if known)

· Species (if known)

· Sex of animal (if known)… see stranding form.

· Location of stranding (e.g. Killiney Strand, Killiney, Co. Dublin)

· Latitude & longitude of stranding location (if known)

The use of a pencil is preferred, in the case of sample tube perforation or breakage, all pen ink (exception of Indian ink) on any paper will run on contact with ethanol. Pencil will not.

6. The Data card carries a unique Sample Number (e.g. 016). Please be careful to place the sample tube in the correct bag with the correct filled in record card, if sampling from more than one individual animal. All sample numbers are cross-referenced to the IWDG Stranding Recording Scheme and the NMINH registry number.

NB: For each stranding, an IWDG Cetacean Stranding form should also be filled out (en