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PROTOCOL FOR COLLECTING TISSUE

This protocol can be downloaded as a PDF for printing if necessary.

If the owners wish, they can have their rabbit’s body back for burial after the following procedure. It will take 10-20 minutes. Most of the time is spent stitching the incision. Sufficient quantity of formol saline (formalin; 10% neutral buffered formalin)  is required. The tissues that need to be collected and submitted are:

  • Heart and lungs (whole organs)
  • Piece of liver (plus another to be frozen)
  • Whole kidney
  • Piece of, or whole, spleen

The histopathologist is willing to look at any additional tissues that are submitted so any other abnormal tissue of interest is found during post-mortem examination, it can be included and sent with the rest of the samples.

Protocol

  • Find a large necked container ( e.g a clean tub that had tablets in it or a plastic lunchbox). Fill it with 50 to 100mls formol saline to fix the tissue.
  • A camera close to hand is an advantage. Photos of any abnormalities are always useful.
  • Make a midline skin incision from throat to pubis and expose the musculature beneath
  • Free some of the skin from the musculature so that it is away from the incision. This prevents hair entering the body cavity and blood contaminating the fur.
  • To open the abdomen, tent the skin for the initial small nick so that air can enter and the linea alba can be cut without opening the caecum. Extend the incision from xiphisternum to pubis. It can be helpful to incise the muscles following the line of the ribs and pubic brim. This allows the muscles to be folded back to expose more of the viscera without letting them displace too much. However, it is more difficult to repair the incision at the end of the procedure
  • Look at the gross appearance of the viscera and presence of any fluid, blood or ingesta in the abdominal cavity.
  • Is the stomach distended and filled with fluid and gas? If so, look for a small intestinal obstruction especially at the pyloric end of the duodenum. The common causes of obstruction are pellets of impacted fur or neoplasia .
  • Examine the liver and assess its size and colour. Take a representative sample (about 2cm of tissue) and place it in the formol saline for histopathology and collect another 2x2cm sample to freeze (unfixed) and submit for PCR testing for Rabbit Haemorrhagic Disease.
  • Lift the stomach and examine the spleen attached to the stomach by lesser omentum. Remove at least half the spleen and put into the formol saline.
  • Move the guts to the right and find the left kidney (and adrenal gland) Note the size and surface of the kidney. Do the same on the other side. Remove at least one kidney and put into formol saline.
  • Examine the bladder and especially if a neutered female look for adhesions between bladder and uterine/vaginal stump.
  • Make a small nick in the diaphragm to allow the lungs to move away from the ventral thoracic wall and then incise alongside the sternum.
  • Retract the ribs and sternum to open the chest cavity. Note the external surface of heart and pericardium. Look at all the lungs in situ.
  • Incise up the neck to expose the trachea. Open the trachea to the pharynx to check for the presence of a foreign body. Note: The internal surface of the trachea in rabbits is naturally hyperaemic and red in colour.
  • Section the trachea and lift/dissect out the pluck. Examine the heart and lungs grossly and preferably gently palpate the lungs. Are the lungs discoloured, collapsed and reddish/pin in patches, obvious tumour or abscess?
  • Put the whole pluck into the formol saline but keep the cut edge of the trachea held in forceps and trickle formol saline into the trachea. Three to 5 mls is all that is needed to fix the lungs internally.Filling lungs with formol saline
  • Preferably inject a couple of mls of formol saline into the chambers of the heart, especially if the heart is large. Then immerse the whole pluck in formol saline.
  • Repair the incision. Paper towelling can be used to pack the chest cavity and absorb any blood

Store the piece of unfixed liver in a sealed container in a freezer until postage instructions are received. Once the other tissues have fixed  ( more than 48 hours), the formol saline can be drained off and the fixed tissues placed in a sealable plastic bag. This bag is then surrounded by paper towels, placed into another sealable bag and finally a padded envelope. This seems to sufficient protection for the tissues. Enclose the lab form and submit to the lab.