Bioassay-guided fractionation of antifouling compounds using computer-assisted motion analysis of brown algal spore swimming.
Antifouling extracts from the sea stars Astropecten articulatus and Luidia clathrata and from the brittle star Astrocyclus caecilia were fractionated by solid phase extraction and high performance liquid chromatography. Bioactive fractions were identified with the use of computer-assisted motion analysis-based bioassays utilising previously described Hincksia irregularis spore swimming behaviour parameters. Quantified parameters of spore movement were rate of change of direction (RCD) and speed (SPEE). The methods used initially required only 10 microg equivalent amounts of total crude extract and each resultant resolving step (normalised to 1 mg ml(-1) of crude, unfractionated extract) required far less material. Statistical analyses of RCD and ratios of RCD:SPEE values in experiments comparing swimming in the presence of extract fractions to controls revealed that both parameters were useful individually and in combination for efficiently following compound bioactivity throughout the fractionation procedure. This technique was also able to detect synergistic or additive interactions between compounds.
Greer
Stephen P
SP
Department of Biology, The University of Alabama at Birmingham, Birmingham, Alabama 35294-1170, USA.
Iken
Katrin
K
McClintock
James B
JB
Amsler
Charles D
CD
D001681:Biological Assay; D002465:Cell Movement; D002851:Chromatography, High Pressure Liquid; D003201:Computers; D021621:Imaging, Three-Dimensional; D009068:Movement / Q000187:drug effects; D000459:Phaeophyceae / Q000166:cytology* / Q000187:drug effects* / Q000502:physiology; D013170:Spores / Q000502:physiology*