Lucifensin II, a defensin of medicinal maggots of the blowfly Lucilia cuprina (Diptera: Calliphoridae).

A novel homolog of insect defensin, designated lucifensin II (Lucilia cuprina Wiedemann [Diptera: Calliphoridae] defensin), was purified from hemolymph extract from larvae of the blowfly L. cuprina. The full-length primary sequence of this peptide of 40 amino acid residues and three intramolecular disulfide bridges was determined by electrospray ionization-orbitrap mass spectrometry and Edman degradation and is almost identical to the previously identified sequence of lucifensin (Lucilia sericata Meigen defensin). Lucifensin II sequence differs from that of lucifensin by only one amino acid residue, that is, by isoleucine instead of valine at position 11. The presence of lucifensin II also was detected in the extracts of other larval tissues, such as gut, salivary glands, fat body, and whole body extract.

Parturition hormone in the tsetse Glossina morsitans: activity in reproductive tissues from other species and response of tsetse to identified neuropeptides and other neuroactive compounds.

Parturition hormone (PH) activity is present not only in the uterus of the tsetse Glossina morsitans but also in the oviducts of Bombyx mori and Schistocerca gregaria, as well as the ejaculatory duct of S. gregaria males. Activity thus appears to be present in the reproductive ducts of diverse insect taxa. To determine whether any of the common insect neuropeptides are capable of mimicking the effect of PH, 35 identified neuropeptides and analogs were evaluated for PH activity. Modest PH activity was observed for only high doses of proctolin and a pyrokinin analog, thus suggesting that PH is unlikely to be closely related to any of the identified neuropeptides tested. While proctolin was highly effective in stimulating contractions of the S. gregaria oviduct, the extract from the tsetse uterus elicited only a weak response in this bioassay. PH activity was, however, effectively mimicked with an injection of 8 bromo-cyclic GMP, thus suggesting a potential role for this cyclic nucleotide in mediating the PH response. Pregnant females were responsive to PH, other neuropeptides and cyclic nucleotides only when females were neck-ligated. In intact females, the brain can presumably override the stimulation provided by the active compounds.

A hormone from the uterus of the tsetse fly, Glossina morsitans, stimulates parturition and abortion.

Unlike most insects, the tsetse female gives birth to a single, fully grown larva at the culmination of each pregnancy cycle. The expulsion of the larva is regulated by a hormone present in rich abundance within the female's uterus. The hormone elicits parturition when injected into neck-ligated females at late stages of pregnancy and abortion when injected at earlier stages. We refer to this highly active material (0.043 uterus equivalents stimulates parturition in 50% of the females) as parturition hormone. Injection of the active extract, which appears to be a peptide or small protein, initiates the series of blood pressure pulsations and uterine contractions normally associated with parturition. The discovery that a uterus extract from the flesh fly also elicits parturition in tsetse suggests that this hormone may be widely distributed in insects.

Neural regulation of pupariation in tsetse larvae.

A neural mechanism coordinates pupariation behavior and tanning in the tsetse larva. At parturition, the mature larva has already received sufficient ecdysteroid to commit the epidermal cells to metamorphosis but, before sclerotization and tanning of the cuticle can begin, the larva must first select a pupariation site and then proceed through a stereotypic sequence of pupariation behavior that culminates in the formation of a smooth, ovoid puparium. Both pupariation behavior and tanning are inhibited by the central nervous system (CNS) during the wandering phase. This central inhibition is maintained by sensory input originating in the extreme posterior region of the body. At the transition from wandering to pupariation, the posterior signal that induces inhibition of pupariation behavior is removed and the larva begins the contractions associated with pupariation, but the CNS inhibition of tanning persists. At this point, separation of the body into two halves by ligation or nerve transection prevents tanning of the anterior half (containing the CNS), whereas the denervated integument of the posterior half tans completely. Transection of nerves to the midline of the body produces larvae with a tanning pattern that ends abruptly along a sagittal plane, implying that the central control of this process is uncoupled between the left and right regions of the CNS. A few minutes later, when the final shape of the puparium is completed, the CNS inhibition is lifted and the tanning process begins. At this time, separation of the body into two halves by ligation or nerve transection has no inhibitory effects on either part. Exogenous ecdysteroids fail to release the CNS inhibition, and hemolymph containing the pupariation factors from Sarcophaga bullata have no accelerating effects on tsetse pupariation. These results imply that regulation of metamorphosis in the insect integument is not the exclusive domain of blood-borne hormones.

Effects of the bipyridylium herbicides diquat dibromide and paraquat dichloride on growth and development of Neobellieria bullata (Diptera: Sarcophagidae) larvae.

Diet containing diquat dibromide (1,000 or 2,000 ppm) caused an extension of the first-instar stadium of Neobellieria bullata (Parker); the first molt was primarily disturbed. Pupariation was delayed when early-wandering larvae had been injected with diquat dibromide (18 micrograms/larva; approximately to 150 ppm). This effect of diquat dibromide was eliminated by simultaneous injection of 20-OH ecdysone (0.02 micrograms/larva). After larvae in the red spiracle stage were injected with diquat dibromide (5 micrograms/larva; approximately 42 ppm), evagination of the pupal head was inhibited. Paraquat dichloride was less active than diquat dibromide and appears to be a safer herbicide for use around decomposer fly species.

Male confusion of the nun moth with disparlure at high and low population densities.

The feasibility of using the synthetic sex pheromone, disparlure, as a mating confusant of the nun moth (Lymantria monacha L.) was investigated on wild populations occurring in outbreak proportions in Poland and in low density in Czechoslovakia. The size of experimental plots ranged from 1 to 12 hectares. The amounts of pheromone applied were 10 or 20 g/hectare. Three types of slow-release formulations of a racemic mixture of disparlure were tested: (1) a spray formulation with latex as a carrier, (2) a tubing of natural rubber, and (3) sawdust of a porous material used for making tips of fiber-tip pens. All three pheromone formulations prevented significant numbers (98.6-100% in the low density situation, 90-96.2% during the outbreak) of the males from locating a discrete source of pheromone (a trap containing synthetic pheromone or a female) during the whole flight period, indicating long-lasting efficiency of the formulations.

Overt and covert effects of endogenous and exogenous ecdysone in puparium formation of flies.

Mature larvae of Sarcophaga argyrostoma fail to form puparia when kept in contact with water, but pupariate after having been subsequently exposed to dryness for 30 hr. They become increasingly more sensitive to injected ecdysone the longer the exposure to dry conditions. A second wet treatment halts the production of hormone that had occurred during the dry period, and reduces a sensitization to injected ecdysone during the intervening dry treatment. Apparently, injected ecdysone had cumulative effects not with the endogenous or exogenous hormone, but with its covert effects which had arisen during the dry period and persisted throughout the second wet period. The hind parts of larvae ligated at the end of a wet period pupariate when injected with either one massive dose of ecdysone, or a far smaller total of several subsequent doses. In the case of two unequal doses, the effect was greater when the smaller dose was injected first. In order to get pupariation with minimal effective doses, a continuous supply of the hormone during a certain period is required. It is suggested that the inhibition of pupariation by moisture has arisen as an adaptation to unfavorable conditions.

Correlated effects of ecdysone and neurosecretion in puparium formation (pupariation) of flies.

A neurohormone accelerates the formation and tanning of the fly puparium in the presence of ecdysone. When larvae of Phormia regina are divided by ligation into two parts before the critical period, the hind part remains untanned, unless injected with blood from a pupariating larva. This confirms earlier observations and is in contrast to recent data with another genus. When Sarcophaga bullata larvae are ligated after the critical period, the anterior part starts to tan about two hours before the posterior part. Injecting neurosecretory material from the brain, the pars intercerebralis, or the corpora cardiaca of S. bullata, or from the brain or c. cardiaca of Periplaneta or Pyrrhocoris into the posterior part induces it to tan before the anterior part. Injection of the same material into normal larvae after the critical period similarly accelerates puparium formation and onset of tanning. Injection of ecdysone or ecdysterone does not have these effects. Thus, the neurosecretory material does not act by causing the release of ecdysone from the ring gland and its function is different from that of the classical brain ("activating") hormone. When injected before the critical period together with ecdysone it potentiates the effect of ecdysone. It is suggested that its biological significance is to coordinate puparial contraction and tanning.