The further application of MTT-formazan colorimetry to studies on filarial worm viability.
Experiments have confirmed that MTT-formazan colorimetry in its simplest form (incubation of intact worms with MTT and direct visualisation of any formazan formed) can be readily applied to several species of filariae including Onchocerca volvulus. Data is presented which will assist the development of quantitative MTT reduction viability tests for a selection of the smaller filarial species. Assays of pieces of Onchocerca gutturosa and O. volvulus females have led us to tentatively conclude that the tips of filariae, particularly the anterior ends, may well be metabolically the most active part of the worm. Selective sampling of these regions for Onchocerca might therefore be a useful indicator for the viability of the parasite. An example of how MTT-formazan colorimetry has been applied to yield additional data to support motility observations on the in vitro survival of male O. gutturosa is also given. The in vitro timecourse of worm death caused by 10 microM CGP 20376 on Acanthocheilonema viteae females has been examined by MTT reduction and compared with 6 other non-subjective parameters. The results suggests that the parameters examined could be divided into two groups according to the time taken for CGP 20376 to cause 50% inhibition (t50) of the parameter. Fast response parameters had t50's between 1 and 6 h (motility indices, 14CO2 evolution, adenine uptake and leucine uptake), they are more sensitive measures of viability and indicate possible worm damage which may or may not be reversible. Slow response parameters had t50's between 34 and 48.5 h (lactate output, MTT reduction and adenine leakage), and are probably linked with severe degenerative changes and are indicative of worm death.(ABSTRACT TRUNCATED AT 250 WORDS)
Comley
J C
JC
Department of Molecular Sciences, Wellcome Research Laboratories, Beckenham, Kent, U.K.
Townson
S
S
Rees
M J
MJ
Dobinson
A
A
D000818:Animals; D002009:Brugia / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism; D003124:Colorimetry; D004396:Coloring Agents; D004153:Dipetalonema / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism; D005260:Female; D005369:Filaricides / Q000494:pharmacology; D005370:Filarioidea / Q000187:drug effects / Q000254:growth & development* / Q000378:metabolism; D005562:Formazans / Q000378:metabolism; D007700:Kinetics; D008297:Male; D009854:Onchocerca / Q000187:drug effects / Q000254:growth & development / Q000378:metabolism; D010084:Oxidation-Reduction; D013778:Tetrazolium Salts / Q000378:metabolism*; D013844:Thiazoles / Q000378:metabolism* / Q000494:pharmacology
D004396:Coloring Agents; D005369:Filaricides; D005562:Formazans; D013778:Tetrazolium Salts; D013844:Thiazoles; C051745:CGP 20376; C022616:thiazolyl blue