Estimation of lipid reserves in unstained living and dead nematodes by image analysis

During storage, non-feeding stages of entomopathogenic nematodes become visibly more transparent due to depletion of energy reserves. Optical density per unit area (OD per area) of infective juveniles of Steinernerna carpocapsae (All) and two Heterorhabditis isolates (UK211 and HF85) was measured with an image analysis system and compared with neutral lipid levels obtained by Oil Red O staining. Optical density (OD) measurements were compared with triglyceride levels of UK211 and HF85. Good correlations between OD per area and neutral lipids (0.90) and between OD and triglycerides (0.87) were found. Thus, OD reflects lipid levels and can be used as an indicator of lipid reserves in these nematodes. Heat-killing of nematodes had no significant effect on OD measurements, but length increased significantly. Storage in a triethanolamine in formaldehyde solution decreased the OD and OD/area by about 5% to 8%. An additional advantage of the image analysis method described is that repeated measurements can be performed on live nematodes. Key words: entomopathogenic nematode, Heterorhabditis, image analysis, neutral lipid, Oil Red O, optical density, Steinernema, triglyceride.