Shearwater Necropsy Lab

Necropsies – Safety

• 70% Ethyl Alcohol: to preserve samples

- Use eye protection

- Eye rinse station

• Sharps: devoted container

- Remove scalpel blade with tweezers

- Cuts: first aid kit

Necropsies – Safety

• P.P.E.: closed – toed shoes, gloves, glasses (alcohol)

• M.S.D.S.:

• Biosecurity: lab coat, gloves, wipes

➢ Manage Risk: follow simple rules and be prepared

➢ Be Respectful: to your class-mates and the birds

Necropsy Manuals • Van Franeker, J.A., 2004. Save

the North Sea Fulmar-Litter-EcoQO Manual Part 1: Collection and dissection procedures. Wageningen, Alterra, Alterra-rapport 672.

http://www.pelagicos.net/pdfs/necropsy/ AlterraRapport_0672_withcover.pdf

• AVIAN NECROPSY MANUAL FOR BIOLOGISTS IN REMOTE REFUGES by THIERRY M. WORK, DVMUSGS, National Wildlife Center, Honolulu

http://www.pelagicos.net/pdfs/necropsy/ USGS_seabirdmanual.pdf

➢ Bring your bird to “measurement” station:

➢ Measure the following:

• Culmen length / depth• Wing length (relaxed)• Tarsus length• Head length• Body mass

➢ Record Data on Necropsy Sheet:

Necropsies – Morphometric Measurements

NOTE: always use metric system (grams / millimeters)

Necropsies – Morphometrics

➢ How to Make Wing Chord Measurements

NOTE: Record if the right wing is fully grown:

Record the following:“1”: P9 is longer than P10“2”: P9 and P10 equal length“3”: P9 is longer than P10

NOTE: we are making the relaxedwing chord measurement (A)

Necropsies – Morphometrics

➢ How to Make Tarsus Length Measurements

NOTE: we are making the “short” tarsus measurement (A)

Necropsies – Morphometrics

➢ How to Make Bill / Culmen Measurements

NOTE: we are also making the “bill depth at base” measurement

Making Wing Measurements

➢ Weight (to the closest gram):

➢ Lay out bird on its back and stretch the wings out.

Making Wing Measurements

Inside of the wing outline, write the following:

- Bird ID (WTSH 2017-100)

- Bird Mass (to the closest gram)

Once your data sheet is complete and checked out:

• Place cotton ball in bird’s mouth

• Bag your bird

• Staple ID in outside of the bag

• Place in “dead bird” tub

Necropsies – External Observations

Next week we will get into the birds…

Bring closed – toed shoes. Bring own lab coat, if you have one.

For each bird, we have two cryovials and two bags:

➢ 2 Cryovials for isotopes: Muscle / Liver• Label with pen on blue painter’s tape• Write down Bird ID and tissue: Liver OR Muscle

➢ 2 Whirl Packs: Feathers / Isotopes• Label whirl pack with black sharpie• Add piece of write-in-the-rain paper• Write down Bird ID and: ”Isotopes” OR “Feathers”

➢ Take 4 – 5 breast feathers, place in “feather bag”➢ Place “feather bag” into “isotope bag”➢ Place cryovials into “isotope bag”

Necropsies – Preparing Vials

Necropsy Steps: Remove Skin

Start by making a lengthwise ventral section just through the skin over the breastbone and belly to near the cloaca.

Attempt to keep the tissue-lining around the intestinal cavity intact.

The thin-walled stomach lies immediately below and can be easily damaged if you cut too deep. Things may not only get messy, but part of stomach contents may get lost… and then there is the GREEN BOMB!!!

Necropsy Steps: Remove Skin

Open the bird by ‘peeling off’ the skin to both sides using your hands and the scalpel.

If you use the scalpel, keep the fat layer attached to the skin.

Check the bird’s condition:

• breast-muscle

• fat stores

• internal molt of the breast coverts

Necropsy Steps: Score Condition

• Record and add three metrics.

• Describe overall condition index it is using these score categories:

0-1 = mortally emaciated; 2-3 = critically emaciated; 4-6 = moderate body condition; 7-9 = good body condition.

Adult Shearwater with Predator Damage

Condition Scores – Pectoral Muscle

Notice the color of the breast muscle: pink or red

Notice internal molt:

0 = none, 1 = one area, 2 = multiple areas

Condition Scores – Subcutaneous Fat

Notice fat around the intestines

Condition Scores – Subcutaneous Fat

Condition Index

Score 2 Fats & Muscle:

3 is BEST

0 is WORST

Add Scores to CalculateOverall Condition Index

(from 0 to 9)

Adult Shearwater in Good Condition

Necropsy Steps: Sample Isotopes

• Wipe blade / tweezers with alcohol after taking samples

• Ensure paper inside of vials

• Store samples in “Isotopes baggie”:

• Feathers• Muscle• Liver

Isotopes: Sample Pectoralis Muscle

• Two sets of muscles: The supracoracoideus lift the wing while the pectoralis provide the powerful downstroke

Largest muscles are the pectorals, or the breast muscles, which make up about 15 -25% of a bird’s body weigh

Body Wall

➢ Notice color and size

• Healthy liver: large, red, bloody, shinny• Unhealthy liver: brown, dry, crumbly

Isotopes: Sample Liver

Necropsy Steps: Check out Organs

• Carefully cut the connective tissue linking the gut and the stomach and the organs under ribs (liver, heart)

Necropsy Steps: Check out Organs

• Air sacs: Provide flotation and cushioning (impacts)

• Lungs: Located on either side and dorsal

Necropsy Steps: Remove Rib Cage

Carefully cut air sacks and the connective tissue lining the stomach, liver and intestines

Goal is to remove the rib cage, so we can look underneath

Necropsy Steps: Remove Rib Cage

Use bone shears to cut ribs on both sides and the scapula

Remove rib cage entirely to expose the organs: liverheart lungsstomach

Necropsy Steps: Score OrgansNotice asymmetry in the organs: liver and stomach

Now carefully cut the lining around stomach/intestines to start inspecting intestinal fat and various organs for e.g. sex determination.

NOTE: Do NOT remove the stomach until you have completed ALL the items from the dissection form.

Organ Scores

Score 5 Organs: 3 is BEST 0 is WORST

- Liver, Gut, Kidney, Lung- Stomach (after removing and opening stomach)

Add Scores to Calculate Overall Organ Health (from 0 to 15)

NOTE: Decay of specimens will complicate judgement of organ health: attempt to give scores as if bird was ‘fresh’.

Health scores for different organs assist in defining the cause of death or the duration of the dying process.

Organ Scores

Score 5 Organs:

- Liver- Gut- Kidney- Lung- Stomach

3 is BEST0 is WORST

Use %s:

0%: not affected

1 – 50%: somewhat affected

51 – 99%: affected

100%: heavily affected

Determine Sex & Age

Determine Sex & Age Sexual organs close to kidneys; can only be found by pushing overlying organs (stomach and intestines) gently to the side.

Initially look for the sexual organs at the LEFT side, because female birds develop sexual organs (ovarium and oviduct) only at the left side of body.

Males have testes on both sides; yet, their size and color not symmetrical

Determine Sex & Age

Checking out the Bursa

At the end of the intestine, check out the ceca and the bursa de fabricius

Ceca serves as the site of fermentation (e.g. vegetation)

Bursa is used to determine the age class of the birds:

- Pre-breeder- Breeding age

Taking out the Stomach

Start by cutting loose the intestine where it leaves the gizzard.

Using blunt tools or your fingers, loosen the proventriculus from the rear of the breast-cavity and move higher up to the throat.

Taking out the StomachOne you have taken out the stomach, carefully place it in a petri dish.

Remove all organ fragments from stomach lining exterior.

Watch out for explosive bile !

Place stomach into sample jar.

Make sure paper tag inside.

Write bird number on blue tape on outside of sample jar.

Necropsy Steps: Finish Dissection

• Record samples taken in form.

• Label and seal sample vials.

• Ensure information recorded legibly.

• Check out with instructors.

• Bag your bird.

• Staple ID in outside of the bag.

• Place in “dead bird” tub.

Necropsy Steps: Clean Up• Remove scalpel blades with tweezers

• Discard in sharps container

• Wipe counters with antibacterial wipes

• Sweep feathers

• Clean utentils and trays with soap in sink

• Dry utensils and trays with paper towels

• Remove feathers from sink drain

• Wash hands with antibacterial soap

Checking out the GizzardCarefully open gizzard (in half, like an orange) over petri dish.

Insert contents into vial, use alcohol to rinse and carefully scrape items with tweezers.

Label sample (Bird ID):- rain paper (inside)- paper tape (outside)

Record stomach / gizzard contents in the form

(e.g., Plastic? Squid beaks?)