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The daily titers of juvenile hormone and ecdysteroids were determined for workers of the Eastern subterranean termite Reticulitermes flavipes (Kollar) (Isoptera, Rhinotermitidae) following isolation of the workers from soldiers and reproductives. Experiments have demonstrated that isolation leads to biochemical and physiological changes that result in a presolider molt. Juvenile hormone (JH) and molting hormone (MH) titers were determined in hemolymph samples using radioimmunoassay. The identity of the ecdysteroids as predominantly 20-hydroxyecdysone (90%) and ecdysone (10%) was determined by HPLC using pooled extracts of hemolymph.

Adult Schistosoma mansoni from experimentally infected mice and Fasciola hepatica recovered from ovine livers post mortem were analyzed for free and conjugated ecdysteroids by radioimmunoassay, high-performance liquid chromatography monitoring fractions by radioimmunoassay, and by capillary gas chromatography/mass spectrometry (selected ion monitoring). Both species contained ecdysone and 20-hydroxyecdysone as free ecdysteroids and as polar conjugates. F. hepatica also contained a polar conjugate of 2-deoxyecdysone. Evidence of apolar ecdysteroid conjugates was only obtained for F. hepatica. The free ecdysteroid-containing fraction of S. mansoni contained small amounts of unidentified immunoreactive material.

The variations in circulating ecdysteroids and juvenile hormones (JH) in Galleria, from the end of the antepenultimate larval stage until emergence of adults, have been determined. The two hormonal families were extracted separately from the same hemolymph sample and quantified by two radioimmunoassays. Juvenile hormone RIA activity was about 35 nM in larvae of the antepenultimate and penultimate stages. It dropped before each molt and increased thereafter. Moreover, it gradually decreased during the last larval instar. In pupae, it was generally low, but it rose drastically during the late pupal development and in young adults. This rise was very much higher in females than in males. Three different RIA-active compounds were found; they were assumed to be JH-I, JH-II, and JH-III according to their retention times in HPLC. The three compounds were present in almost equal concentration in larvae of the penultimate stage: JH-I predominated, however, during the last larval instar. In late pupae, the main hormone was JH-III both in males and in females. There is no clear relationship between ecdysteroid and juvenile hormone changes, except for a female-specific ecdysteroid rise which coincides with the juvenile hormone release in late pupae. This double hormonal stimulation can be involved in the regulation of vitellogenin synthesis and deposition in oocytes.

Prepatent and patent adult Hymenolepis diminuta from the intestines of rats, H. diminuta eggs recovered from the faeces of rats harbouring patent infections, and infective cysticercoids from the beetle intermediate host were analysed for free and conjugated ecdysteroids. Adult worms and eggs contained both free ecdysteroids and hydrolysable polar conjugated ecdysteroids, with comparatively large amounts of immunoreactive material also being detected following hydrolysis of the possible apolar conjugated ecdysteroid fraction. Free ecdysteroids were not detected in the cysticercoid sample. The concentration of free ecdysteroids in H. diminuta eggs was higher than that detected in the tissues of the adult worms. Ecdysone and 20-hydroxyecdysone were the major identified compounds of the free ecdysteroid fraction, whereas in the hydrolysed polar conjugated ecdysteroid fraction these two compounds were accompanied by 20,26-dihydroxyecdysone. The free ecdysteroid fraction also contained comparatively large amounts of unidentified immunoreactive material.

Blood-fed, decapitated female Aedes aegypti mosquitoes matured eggs when injected with an extract of heads, but non-blood-fed females did not. The response to head extract was optimal when the head was allowed to remain for 2 hr after feeding. A dose response to injected egg development neurosecretory hormone (EDNH) was observed in vivo that was similar to in vitro dose responses previously reported. Blood-fed decapitated females responded equally well to boiled or unboiled head extract. When blood-fed decapitated females were injected with head extract, ecdysteroid levels increased. Partial purification of head extract using high-pressure liquid chromatography yielded a fraction at 34% acetonitrile that showed egg maturation activity in vivo when injected into blood-fed decapitated females, and ecdysiotropic activity when incubated in vitro with ovaries. In addition, a fraction at 30% acetonitrile was found that showed activity in vivo but not in vitro and may be a precursor. Occasionally, the fraction at 37% acetonitrile showed activity in the in vitro assay but had little activity in vivo and may be a metabolite. These results suggest that the same hormone was being assayed in vivo and in vitro and is EDNH.