I will be making blood agar using fish blood. Can anyone give me the following information? There is a paper, but these questions are not answered and I have not been able to contact the authors to get the information.

1. When drawing the blood, should it be drawn into a vaccutainer already containing the anticoagulant (I will be using Alsever's solution mixed 1:1 with the blood, as that was used in the paper)?

2. When 5% blood by volume is mixed into the base agar does one mix packed cells, whole blood, or blood mixed 1:1 with Alsever’s soln. (that is, should the 5%be calculated on the whole blood volume as drawn or on the blood/Alsever’s mixture) ?

If anyone answers who has made fish blood agar, I have the following questions:

1. Have you tried any percentages of blood other than 5%?

2. Have you tried any other types of agar other than TSA as the base for the blood agar (that's what the authors of the paper used)?

3. I'm thinking of trying this with rainbow trout. Have you experience with any species of fish other than tilapia (again, that was the species of fish used in the paper)?

Thank you for your consideration.