TITLE: Testing selective agents for the Icthyosporeans Abeoforma whisleri, Pirum gemata and Spaheroforma artica
AUTHORS: Elena Casacuberta, Cristina Aresté

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span>Minimal protocol for testing selective agents for </span><span style = "font-style:italic;">Abeoforma whisleri</span><span>, </span><span style = "font-style:italic;">Sphaeroforma arctica, Pirum gemmata</span></div></div>

[STEPS]
?. Seed cells on 24 well/plate. Density depends on the organism but should allow to screen for growth and death easilySeed the same density to test four or five different dilutions of each drugSeed enough wells to have 3X replicates for each conditionSeeded cells for Abeoforma whisleri, Pirum gemata and Spaheroforma artica 100 cells/well 500 microliters of medium.
?. Add to each well the corresponding amount of drug. Following the amounts tested in our case for Abeoforma whisleri, Pirum gemmata, Sphaeroforma arctica  AB1Nourseothricin&nbsp;10-50 μgr/ml&nbsp;&nbsp;&nbsp;2Phleomycin&nbsp;10-300 μgr/ml&nbsp;&nbsp;&nbsp;3Fluorotic Acid&nbsp;250-30ugr/ml&nbsp;4Benomyl&nbsp;&nbsp;Carboxine&nbsp;&nbsp;Puromycin&nbsp;&nbsp;&nbsp;1-20&nbsp;μgr/ml&nbsp;&nbsp;&nbsp;&nbsp;20-300μgr/ml&nbsp;&nbsp;&nbsp;&nbsp;100-500&nbsp;μgr/ml&nbsp;&nbsp;&nbsp;
AB1Nourseothricin&nbsp;10-50 μgr/ml&nbsp;&nbsp;&nbsp;2Phleomycin&nbsp;10-300 μgr/ml&nbsp;&nbsp;&nbsp;3Fluorotic Acid&nbsp;250-30ugr/ml&nbsp;4Benomyl&nbsp;&nbsp;Carboxine&nbsp;&nbsp;Puromycin&nbsp;&nbsp;&nbsp;1-20&nbsp;μgr/ml&nbsp;&nbsp;&nbsp;&nbsp;20-300μgr/ml&nbsp;&nbsp;&nbsp;&nbsp;100-500&nbsp;μgr/ml&nbsp;&nbsp;&nbsp;
?. Spin down cells every 24 hours and refresh media with new antibiotic.Observe for up to a week.If effects on growth are suspected repeat experiment with the apropriate concentration of drug and count cells every 24 hours.
?. Summary of our results  ABC1Abeoforma whisleriNourseothricin&nbsp;10-50 μgr/ml&nbsp;&nbsp;&nbsp;(not sensitive)2Phleomycin&nbsp;10-300 μgr/ml&nbsp;&nbsp;&nbsp;(not sensitive)3Fluorotic Acid&nbsp;250-30 μgr/ml&nbsp;&nbsp;&nbsp;(not sensitive)4&nbsp;Benomyl&nbsp;&nbsp;Carboxine&nbsp;&nbsp;Puromycin&nbsp;&nbsp;&nbsp;1-20 (500)&nbsp;μgr/ml&nbsp;&nbsp;&nbsp;&nbsp;20-300&nbsp;μgr/ml&nbsp;&nbsp;&nbsp;&nbsp;100-500&nbsp;μgr/ml&nbsp;&nbsp;&nbsp;(cytostatic)&nbsp;&nbsp;&nbsp;&nbsp;56Pirum gemmataNourseothricin&nbsp;10-50 μgr/ml&nbsp;&nbsp;&nbsp;(not sensitive)7Phleomycin&nbsp;10-300 μgr/ml&nbsp;&nbsp;&nbsp;8Fluorotic Acid&nbsp;250-30ugr/ml (not sensitive)9&nbsp;Benomyl&nbsp;&nbsp;Carboxine&nbsp;&nbsp;Puromycin&nbsp;&nbsp;&nbsp;1-20&nbsp;μgr/ml&nbsp;&nbsp;&nbsp;&nbsp;20-300μgr/ml&nbsp;&nbsp;&nbsp;&nbsp;100-500&nbsp;μgr/ml&nbsp;&nbsp;&nbsp;(cytostatic)10
ABC1Abeoforma whisleriNourseothricin&nbsp;10-50 μgr/ml&nbsp;&nbsp;&nbsp;(not sensitive)2Phleomycin&nbsp;10-300 μgr/ml&nbsp;&nbsp;&nbsp;(not sensitive)3Fluorotic Acid&nbsp;250-30 μgr/ml&nbsp;&nbsp;&nbsp;(not sensitive)4&nbsp;Benomyl&nbsp;&nbsp;Carboxine&nbsp;&nbsp;Puromycin&nbsp;&nbsp;&nbsp;1-20 (500)&nbsp;μgr/ml&nbsp;&nbsp;&nbsp;&nbsp;20-300&nbsp;μgr/ml&nbsp;&nbsp;&nbsp;&nbsp;100-500&nbsp;μgr/ml&nbsp;&nbsp;&nbsp;(cytostatic)&nbsp;&nbsp;&nbsp;&nbsp;56Pirum gemmataNourseothricin&nbsp;10-50 μgr/ml&nbsp;&nbsp;&nbsp;(not sensitive)7Phleomycin&nbsp;10-300 μgr/ml&nbsp;&nbsp;&nbsp;8Fluorotic Acid&nbsp;250-30ugr/ml (not sensitive)9&nbsp;Benomyl&nbsp;&nbsp;Carboxine&nbsp;&nbsp;Puromycin&nbsp;&nbsp;&nbsp;1-20&nbsp;μgr/ml&nbsp;&nbsp;&nbsp;&nbsp;20-300μgr/ml&nbsp;&nbsp;&nbsp;&nbsp;100-500&nbsp;μgr/ml&nbsp;&nbsp;&nbsp;(cytostatic)10