Differences in lipid granulation as the basis for a morphologic differentiation between third-stage larvae of...

Differences in the distribution of lipid granules between unstained third-stage larvae of Uncinaria stenocephala and Ancylostoma caninum cultured at 15 C was found to be an effective means for differentiating these 2 species of canine hookworms. In contrast, larvae cultured at 22 C were less easily differentiated based on the distribution of lipid granules. After culturing at 15 C, third-stage larvae of U. stenocephala were motile and exhibited 32 well-demarcated intestinal cells which contained intracellular lipid granules. Intestinal cells were easily visualized due to the absence of extraintestinal lipid granulation. Ancylostoma caninum third-stage larvae cultured under similar conditions were significantly less motile and contained extraintestinal accumulations of lipid granules which obscured intestinal cells. Both species exhibited an overall decrease in lipid granulation during a 14-day observation period following culture at 15 C. Morphologic differentiation of these 2 species after 14 days was based on the absence of intra- and extra-intestinal lipid in U. stenocephala and the presence of some lipid granules in both these locations in A. caninum. The first- and second-stage larvae of both species cultured at 15 C exhibited dense accumulations of extraintestinal lipid granules and were morphologically indistinguishable. This suggests that the observed difference in lipid granulation between the third-stage larvae of U. stenocephala and A. caninum cultured at 15 C is due to differences in lipid utilization during the third stage rather than differences in lipid synthesis by the first- and second-stage larvae and is related to the adaptation of these parasites to their respective climatic regions.