Direct Inoculation from Kidney

  • Using a scalpel blade, carefully make an incision through the body wall in the mid-ventral line at the base of the pelvic fins.
  • Using scissors or scalpel blade extend the incision anteriorly and posteriorly to expose the internal organs. Take care not to puncture any part of the intestinal tract.
  • Hold, pin back or remove the side of the body wall.
  • Carefully move the alimentary tract to one side to expose the kidney, which is covered by the swim bladder.
  • Carefully peel back the swim bladder.
  • Note any abnormalities.
  • Sterilise a bacteriological loop by heating and allowing to cool or use a disposable loop.
  • Sterilise a scalpel and use this to make a small incision in the kidney (optional)
  • Insert a sterile loop into the kidney and remove a small portion of kidney using the loop, taking care not to touch the loop on any other part of the fish.
  • Inoculate the sample on a segment of the surface of the culture medium. This is known as the well.
  • Sterilise the loop, allow to cool and touch the edge of an ininoculated area of medium to ensure coolness.
  • Spread part of the sample over about 1/4 of the plate by making 3-4 parallel streaks with the loop.
  • Repeat streaking procedure as shown below, sterilising and cooling the loop or using a new disposable loop between each sequence.
  • Label the underside (not the lid) of the plate, writing only round the edge.
  • Seal plate with nescofilm or similar sealing tape before transport.