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.