Identity and tissue localization of free and conjugated ecdysteroids in adults of Dirofilaria immitis and Ascaris suum.

Adult males and females of the dog heartworm, Dirofilaria immitis, and of the swine parasite, Ascaris suum, were extracted, the free and polar conjugated ecdysteroid fractions separated and the latter hydrolysed enzymically. The ecdysteroids released by hydrolysis of the conjugates and the free hormones were analysed by radioimmunoassay, high-performance liquid chromatography on reversed phase and adsorption columns monitoring fractions by radioimmunoassay, and by gas-liquid chromatography/mass spectrometry (selected ion monitoring). In both species, males and females contained free and polar conjugated ecdysteroids, with evidence for the presence primarily of ecdysone and 20-hydroxyecdysone together with smaller amounts of 20,26-dihydroxyecdysone. Males and females of both species were then dissected into body fluid, reproductive system, gut and remaining body wall compartments, the ecdysteroids extracted, fractionated and analysed by radioimmunoassay and high-performance liquid chromatography monitoring fractions by radioimmunoassay. The results for both sexes in the two species were similar and indicated that ecdysteroids were not detectable in body fluids and that free ecdysteroids occurred in the reproductive system and the body wall, whereas polar conjugated ecdysteroids were detected in the reproductive system and the gut; a minor portion of the free ecdysteroids in A. suum was also apparently present in the gut. Further localization of the ecdysteroids in the body wall of A. suum females suggested that negligible immunoreactivity was associated with the circumpharyngeal nerve ring. The possible significance of the results is discussed.

Metabolism and fate of ecdysteroids in the nematodes Ascaris suum and Parascaris equorum.

When injected with [3H]ecdysone and maintained in vitro, the parasitic nematodes, Ascaris suum and Parascaris equorum each produced a series of polar and relatively apolar metabolites. A. suum metabolised the compound into ecdysonoic acid ([3H]EOIC), ecdysone 25-glucoside ([3H]E25gluc), putative ecdysone 22-phosphate ([3H]E22P) and a series of at least six relatively apolar metabolites. All of these, except ecdysonoic acid, were hydrolysed by a crude enzyme preparation from Helix pomatia, releasing ecdysone. In a similar study, P. equorum produced ecdysone 25-glucoside, putative ecdysone 22-phosphate and a series of relatively apolar compounds all of which were hydrolysed by H. pomatia enzymes, releasing ecdysone. [3H]Ecdysone 25-glucoside was the most abundant single metabolite in both species, and in P. equorum, at least, was released into the culture medium in relatively large amounts. Apolar metabolites were present in worm samples and were the major, if not the only radiolabelled compounds detected in eggs of both species. Data indicated a metabolic relationship between some of the apolar conjugates found in both nematode species and ecdysone 25-glucoside.