Re: Nosema sampling and preparation technique

From: Jonathan Dick
Date: 3/9/98
Time: 10:17:05 PM
Remote Name: 198.161.229.180

Comments

Nosema Test

Procedure: The bees were collected from under the lids of hives; two samples were taken from approximately the same place and the same time. They were put in sample jars filled with solution (approximatly 1/4 alcohol and 3/4 water).

I crushed either the whole bees or whole abdomens (see table) in a cleaned bowl using a cleaned spoon. I then added 50 ml of the solution (measuring with a kitchen measuring spoon set) and mixed. Next, I poured the mixture through the fine sieve and into a clean sample jar (pill bottle); I did not filter the mixture; I did not add more water.

The mixtures sat for a few days after our initial attempts with a child’s microscope, waiting on a better one (We had not seen spores on our first few samples, maybe because there weren’t many). I then proceeded.

Immediately after shaking the mixture I took a sample with a 40 mm wire loop (made of frame wire); I push the loop to the bottom of the mixture then back up. It is difficult to know at what mixture level the sample was taken. I assumed that the shaking would make the sample homogenous

We used equivalent of ~1 ml of water per bee.

The formula, from page 7 in the instructions we received:

No. spores counted / 80 * 4*10^6 = Spores per bee

Jonathan Dick