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The large white butterfly Pieris brassicae L. (also called cabbage white) is very common in Europe, Asia and the northern region of Africa, and has also been found in South Africa during approximately the last 20 years. The species is considered a pest insect, with larvae attacking brassicaceous crops. The adult is a strong migratory flyer and new territory can be infested this way. As a first step to investigate methods for combating this pest species, the present study aims to determine the complement of adipokinetic peptides, here generically referred to as adipokinetic hormones (AKHs), which are required to regulate the mobilization of fuels for insect flight. Biological assays, as well as mass spectrometry, reveal information about the presence, structure and function of AKHs in P. brassicae: a methanolic extract of the corpora cardiaca has hypertrehalosaemic activity in cockroaches, does not cause hyperlipaemia in locusts, and has adipokinetic activity in P. brassicae itself. Liquid-chromatography electrospray ion trap mass spectrometry reveals three peptides that can be associated with the AKH family: the non-amidated undecapeptide Vanca-AKH (pELTFTSSWGGK-OH), the nonapeptide Manse-AKH (pELTFTSSWG amide) and the novel octapeptide Piebr-AKH (pELTFSSGW amide). Sequence confirmation of all three assigned structures is obtained from matching mass spectrometry spectra from synthetic and native peptides. Moreover, the synthetic peptides Manse-AKH and Piebr-AKH have significant hyperlipaemic (=adipokinetic) activity when injected into newly-emerged adult cabbage white butterflies. The non-amidated Vanca-AKH is, apparently, incompletely processed Manse-AKH without hormonal activity. Simulated dispersal flight is able to release AKHs, as indicated by the higher concentration of lipids in the haemolymph of adult P. brassicae after activity and rest periods.

To survive, an animal must respond to the environmental stimuli (Zeitgebers) precisely coinciding with a 24-h light/dark cycle. In human habitats, both natural and artificial Zeitgebers intermingle, entraining the circadian clocks of animals. Whether a circadian clock can respond to Zeitgebers effectively is closely related to the level of adaptive competitiveness. In the present review, the German cockroach (Blattella germanica L.) is compared with the double-striped cockroach (Blattella bisignata Brunner von Wattenwyl) and the reasons why B. germanica is a more competitive dweller than its sibling species in human habitats are discussed. For example, sensitive adjustment of circadian clocks allows a male B. germanica to coordinate itself to the changes of daily light/dark cycles more efficiently; by contrast, it takes male B. bisignata more than 3 days to adjust. In a given dark phase, male B. germanica appear more to be resistant than B. bisignata to interference from light pulses. Ovary-dependent masking factors enable female B. germanica to take advantage of the light phase to feed or mate, during which time a female B. bisignata typically remains inert. Furthermore, the gregariousness levels, fertilization and hatching rates, as well as mating behaviours of both cockroach species are compared. All of these comparisons suggest that B. germanica surpasses B. bisignata with respect to adapting to the artificial constraints within human habitats as a result of the adoption of an integrated strategy consisting of circadian and noncircadian approaches.

This work was supported by grants BFU2006-01090/BFI (Spanish Ministry of Science and Innovation (MICINN) and FEDER), BFU2010-15906 (MICINN) and 2014-SGR-619 (Secretaria d'Universitats i Recerca, Catalan Goverment) to JLM. MA and SS-C were recipients of a pre-doctoral fellowship (MICINN) and a post-doctoral contract (CSIC, JAE program co-funded by the European Social Fund), respectively. We thanks the Human Genome Sequencing Center of the Baylor College of Medicine for access to the German Cockroach Genome Project.

The composition of cuticular and internal lipids in females of the cockroach Blatta orientalis L. exposed to the entomopathogenic fungus Conidiobolus coronatus is investigated. The compositions of the fatty acids, n-alkanes, alcohol, sterols and methyl esters in the lipids are chemically characterized. Although contact with virulent colonies of the fungus does not induce insect mortality, significant changes in the lipid profiles, both cuticular and internal, are found. The cuticular extracts of a control group of B. orientalis females contain 24 compounds varying in carbon chain length from C6 to C22. The main cuticular fatty acids identified are: C16:1, C16:0, C18:1 and C18:0. The cuticular lipids of B. orientalis females after exposure to C. coronatus contain only 14 free fatty acids from C8 to C20. The highest concentrations identified are C16:0, C18:2 and C18:1. Analysis by gas chromatography-mass spectrometry identifies the presence of a homologous series of n-alkanes containing from 25 to 31 carbon atoms. In the case of the insects after fungal exposure, the content of the n-alkanes in the cuticular lipid is two-fold higher compared with the controls. Of the cuticular lipids, 11 alcohols are found, ranging from C12:0 to C20:0. There is no presence of alcohols in the internal lipids of the control B. orientalis females and in all of the extracts from the B. orientalis females after fungal exposure. In the samples analyzed, the most common sterol is cholesterol. This is present in the cuticular lipids and the internal lipids of all of the insects sampled. The cuticular and internal lipids of females contain five fatty acid methyl esters, ranging in size from C15 to C19.

Neuropeptides are important with respect to almost all physiological processes and behavioural patterns in an organism. In the present study, muscle bioassays, immunohistochemistry and matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) mass spectrometry are used to investigate the distribution and efficacy of proctolin in the antennal heart of 36 species of lower Neoptera. In total, 20 species of Dictyoptera (cockroaches, termites, praying mantids), eight species of Saltatoria (crickets and grasshoppers) and eight species of Phasmatodea (stick insects) are investigated. The antennal heart of all tested Blattoidea, including the termite Mastotermes darwiniensis Froggatt, exhibit a strong proctolin-like immunoreactivity, as well as a high sensitivity to proctolin, whereas members of the second clade of cockroaches (Blaberoidea) are largely insensitive to proctolin and show only a weak proctolin-like immunoreactivity. The antennal heart of praying mantids (Mantodea) also contains only few proctolin-like immunoreactive fibres but is highly sensitive to proctolin. Such a high sensitivity is found also in Phasmatodea, although the antennal heart of these insects does not have proctolin-like immunoreactive fibres. These findings are supported by MALDI-TOF mass spectrometry; no trace of proctolin is detected in antennal heart preparations of Phasmatodea. In Saltatoria, only weak (or no) effects of proctolin are observed and, in most subtaxa, no proctolin-like immunoreactivity is visible in the antennal heart preparations. Only in Grylloidea is strong proctolin-like immunostaining found in processes of the antennal heart. In these species, weak or moderate effects of proctolin are observed in the antennal heart bioassay. In general, the differences in distribution of proctolin and effects on the antennal heart within the basal Neoptera cannot be deduced from their phylogenetic position, although they show conformity within each (sub)taxon.

Crowding causes many organisms to express phenotypic plasticity in various traits. Phase polyphenism in desert locusts represents one extreme example in which a solitary form (solitarious phase) turns into a gregarious form (gregarious phase) in response to crowding. Conspicuous differences in body size and colour occur even in hatchlings. The phase-specific differences in hatchling characteristics are caused by the tactile stimuli perceived by the antennae of their mother. However, the nature of the tactile stimuli and the mechanism by which the perceived stimuli are processed as a gregarizing signal remain unknown. To explore this problem, the antennae of solitarious adult females of the desert locust Schistocerca gregaria are touched with the bodies of conspecific locusts at different physiological stages and those of other species. The results suggest that a cuticular chemical factor at a specific developmental stage of conspecific locusts causes the solitarious females to produce large eggs that give rise to black hatchlings characteristic of gregarious forms (progeny gregarization), and that this or a similar compound occurs in other acridids, crickets and cockroaches but not in beetles. The involvement of a chemical substance is also supported by hexane extracts of cuticular surfaces of locusts that induce the same effects. Interestingly, crowding induces such gregarizing effects only when the female receives the appropriate stimulus in the presence of light. Solitarious female S. gregaria with their head capsule coated with phosphorescent paint exhibit progeny gregarization in response to crowding and light pulses in darkness, whereas those treated in the same way without light pulses fail to do so.

Proctolin is a neuroactive pentapeptide first isolated from the cockroach Periplaneta americana in which it has an excitatory effect on contractions on visceral muscles of the hindgut. Subsequently, proctolin is reported in a wide variety of invertebrates, and considerable efforts have been made to determine its mode of action. Its primary role appears to be that of a neuromodulator rather than a classical neurotransmitter, and it may also serve as a neurohormone, depending on the muscles examined. The present study identifies the vagina muscles of the blood-sucking insect Rhodnius prolixus (Stal) as a proctolinergic system. Physiological doses of proctolin generate prolonged contractions that closely mimic the effects of motor nerve stimulation. This preparation is convenient and robust, warranting its use as an experimental system to further understand the role of proctolin in the regulation of muscle contractions in insects. Moreover, these muscles are innervated by an identifiable inhibitory component providing a means to investigate the interaction between proctolin excitation and neural inhibition.

Electromyographic recordings were made from muscles that extend the trochanter/femur of each of the six legs of American cockroaches, Periplaneta americana (L.), while the insects swam in water. The recordings showed two novel features. (1) During swimming, muscle activity in different legs was coordinated in the alternating tripod pattern commonly seen during free walking on land, not in the pattern of synchronous leg pairs common to other large terrestrial insects in water. (2) Fast axons were usually recruited along with slow axons, even when the insect swam at a moderate pace. Fast axon activity always started after the middle of the slow axon burst in intact insects, but vanished from most bursts in the stump of the leg after amputation of the femur. The alternating tripod pattern was maintained even after amputation. Possible causes of fast axon recruitment are discussed.

Orally supplied octopamine significantly enhances the wing-raising behaviour in male American cockroaches Periplaneta americana, in response to periplanone B the main component of the sex pheromone, but not attraction. In addition, octopamine decreases the response to the general odorant, hexan-1-ol. The presence of a calling female, as well as octopamine treatment, eliminates the avoidance of hexanol by males. Electroantennogram responses to hexan-1-ol decreases after topical octopamine application.

A review of the literature on the circadian activity rhythms in cockroaches led Vancassel (1968) to suggest that both the eyes and the ocelli play a role in the induction of rhythms in previously arrhythmic cockroaches. Experiments involving eye-opaquing or cauterizing techniques were performed to test this hypothesis. The results imply that both kinds of photoreceptors are involved in the setting up of a rhythm, and that light transmitted via the peri-antennal cuticle may be as well. The role of these forms of photoreception in the entrainment of free-running circadian rhythms was not examined.

Carbon dioxide anaesthesia differentially affects the knockdown and recovery of cockroaches, depending on the strain and the length of time that the colony has been subjected to a CO2 regime. Adult males from two laboratory and two field-collected strains of German cockroaches, Blattella germanica, are knocked down within 7–45 s after exposure to CO2. After 5 min of CO2 exposure, presumptive recovery (i.e. the time for the cockroach to right itself after knockdown) for laboratory strains occurs significantly sooner than for field-collected strains. Control cockroaches, exposed to compressed air rather than CO2, exit harbourage cups rapidly (≤3.20 min). However, although allowed a recovery period of 5 min, significant movement impairment occurs for all cockroach strains anaesthetized with CO2. Carbon dioxide exposure significantly reduces consumption of 2.15% hydramethylnon bait and delays mortality even when 24 h is allowed for recovery before bait placement. Cockroaches allowed to recover for 48 h after 5 min of CO2 exposure consume significantly more bait and die significantly faster than CO2 exposed groups allowed 24 h of recovery, and mortality is not significantly different from nonanaesthetized bait-fed controls.

The effects of photoperiod on nymphal growth and adult reproduction were investigated in a small cockroach, Margattea satsumana, living on the subtropical, Hachijo island (33°N), Japan. Nymphal development is slow under constant photoperiods at 25 °C. The shortest mean duration of nymphal development (176 days) is observed at LD 14 : 10 h, followed by LD 12 : 12 h (221 days) and LD 16 : 8 h (309 days). Nymphal development is further prolonged when the nymphs are transferred from LD 12 : 12 to LD 16 : 8 h at 90 days after hatching. However, rapid and synchronized development is observed when nymphs were transferred in the opposite direction. A decreased change in photoperiod from LD 14 : 10 to LD 12 : 12 h also reduces the duration of nymphal development, and this cannot be explained by the results obtained at constant photoperiods. Similarly, nymphs reared at LD 16 : 8 h during the first 60 days mature more rapidly when transferred to LD 12 : 12 h than when transferred to LD 14 : 10 h. The developmental suppression induced by long days may represent a form of summer diapause that is terminated rapidly by short days. Based on these observations and field-census data, it is suggested that this cockroach has a univoltine life cycle overwintering as nondiapause adults, and that this life cycle is stabilized by the response to changing photoperiod.

Diapause is common among insects and is regarded as an adaptive response to periodic occurrence of adverse conditions. It occurs at a particular developmental stage, typically only once in a lifetime. However, little is known about the details of the control mechanism of life cycles with multiple diapauses in insects. In this study, a complex 2-year life cycle with three types of diapause is reported in a subtropical cockroach, Symploce japonica: a winter diapause in mid-nymphal instars, a summer diapause in later nymphal instars, and a reproductive diapause is reported in the adult stage. Nymphal development was extremely slow either at short days (winter diapause) or long days (summer diapause). Nymphs in summer diapause matured rapidly when transferred from long days to short days, indicating that seasonal changes in day-length are the pivotal factor in the control of this life cycle. It is proposed that the main significance of winter diapause in this subtropical species is to enable the nymphs to survive the mild winter successfully with reduced energy demand, and that of summer diapause is to delay adult emergence until late in the autumn for successful induction of the following adult diapause. Adults do not emerge until shortly before winter, yet the presence of diapause in the adult stage does not simply appear to be a response to cope with the winter conditions but, instead, ensures that reproduction will occur early the next year, before summer, because reproduction is greatly hampered at high temperature.

Developmental, reproductive and size-related variables were compared between two ecologically different strains of Periplaneta americana (L.). One strain was a laboratory-reared culture, and the other a feral strain inhabiting an urban environment. The feral population was originally founded by escapees from the culture, but may also include immigrants from other urban populations. Both final-instar nymphs and adults of the feral insects were heavier than their equivalents from the cultured population, this weight difference being due to a higher capacity for the storage of water and heavier fat-free carcasses. Feral cockroaches also had heavier oothecae, which contained heavier offspring than those from cultured females. Feral animals had one or two more larval stadia and higher growth rates. Size-related differences persisted in first and second filial generations reared under laboratory conditions on a nutritionally balanced diet, but were not apparent in first filial insects reared on a vegetable diet. Greater resistance to starvation was found in feral animals, and this was attributed primarily to their larger water stores. Feral animals were found to harbour a higher density of endosymbiotic bacteria in the fat body, which are known to enhance the efficiency of protein utilization. The data suggest that the characteristics of feral animals have been selected in the nutritionally harsh feral environment compared with the more benign culture conditions, with water availability playing a role.

Summary A monoclonal antibody to allatostatin I of the cockroach Diploptera punctata was used to establish a competitive enzyme-linked immunosorbent assay for quantification of allatostatin-like peptides in the hindgut of the adult male earwig, Euborellia annulipes. Hindguts of 0-day males contained significantly more allatostatin-positive material than those of 8-day males fed on catfood. However, males starved for the first 8 days of adult life had significantly higher levels of allatostatin-positive material than those of either 0-day or of 8-day fed males. Hindguts from 0-day old males exhibited lower spontaneous motility in vitro than those from 8-day males. Hindguts from males at both ages responded to allostatin with reversible, dosage-dependent decreases in hindgut motility, and responded to proctolin with reversible, dosage-dependent increases in hindgut motility. When both allatostatin and proctolin were applied to hindgut preparations simultaneously and in equal concentrations, the response varied with the stage of the male. Starvation enhanced hindgut motility and abolished the response to allatostatin, but not to proctolin. These results indicate the presence of material similar to cockroach allatostatins in male earwigs, and that the levels change with age and physiological stage. Furthermore, such peptides may indeed be regulatory neuropeptides and could modulate hindgut contraction. There was an increase in sensitivity to exogenous allatostatin in the hindgut during development from day 0 to day 8 in feeding males, but a loss in sensitivity in response to starvation; sensitivity to exogenous proctolin also increased with age, but such responsiveness was not diminished by starvation.

Summary Allatostatins with a typical C-terminal sequence YXFGL-NH2 are insect neuropeptides with inhibitory properties upon Juvenile Hormone production in the corpora allata, vitellogenin release by the fat body, and gut and dorsal vessel motility. All these biological effects are rapidly reversible, suggesting the occurrence of effective mechanisms for inactivation of the peptides. We have studied the degradation of DRLYSFGL-NH2 (BLAST-2), one of the allatostatins of Blattella germanica, in the internal milieu of adult females of this cockroach. The experimental approach combined the use of the radioiodinated derivative [125I-Tyr4]BLAST-2, microdialysis techniques and HPLC analysis with a radioisotope detector. Under these experimental conditions, the half-life of BLAST-2 in the internal milieu of the adult female of B. germanica was between 3 and 6 min. Such a short half-life explains the high doses of allatostatins required to obtain the expected biological effects when tested in vivo, and suggests that circulating allatostatins are subject to rapid rates of synthesis and degradation in order to be operative physiologically.

Summary In adult female cockroaches, the ovary greatly affects the synthesis of Juvenile Hormone (JH) by the corpora allata, and in females of some cockroach species, removal of the ovaries results in a permanent depression of JH synthesis. We report that the corpora allata in ovariectomised, adult virgins of the German cockroach, Blattella germanica (L.), increase and then decrease in activity, as they do in intact females. Moreover, the distal tubules in the left colleterial glands of ovariectomised females accumulate abundant protein, the production of which is regulated by JH. In both ovariectomised and sham-operated females, the activity of the corpora allata more than tripled between days 1 and 4 of adulthood, during which the oocytes of sham-operated females grew considerably in length. The corpora allata of sham-operated females produced even more JH on day 7, but very little on day 10, by which time all females had oviposited. The glands of ovariectomised females, by constrast, produced a similar amount of JH on day 7 as on day 4, but much less on day 10. Beginning on day 13, the activity of the corpora allata increased again in ovariectomised females, an increase that did not occur until day 22 in sham-operated females. Mating of ovariectomised females on day 6 resulted in a significant increase in the activity of the corpora allata by day 10. We conclude that both the ovary and mating stimulate the synthesis of JH early in the reproductive cycle, but that neither is needed for the occurrence of a complete cycle of JH synthesis.

Summary Blattella bisignata (Brunner) and B. germanica (L.) are oviparous cockroaches with cyclic reproductive behaviour, but in B. germanica only males show circadian rhythmicity of locomotion at 28°C and DD (constant darkness). In B. bisignata, males and virgin females cockroaches entrained by light–dark cycles show free-running rhythmicity in DD, and most activities occur during the subjective night. Daily locomotor activities of virgin females show cyclic changes that coincided with ovarian development. Virgin females also exhibit calling behaviour during the subjective night, and this shows a free-running rhythm. Male mate-finding locomotion and female calling behaviour are under circadian control, so the timing for both behaviours is synchronized. However, most mated females do not show a locomotor free-running rhythm under DD conditions. Our results indicate that only mated females could not express a circadian locomotor rhythm. Pregnancy reduces a female’s locomotory intensity and masks the expression of a circadian locomotor rhythm. We attribute the differences in circadian locomotory rhythms between these two species to their living environments and mate-finding strategies.

Corpora cardiaca of the earwigs Labidura riparia and Forficula auricularia contain a substance that causes hyperlipaemia in migratory locusts and hypertrehalosaemia in the American cockroaches. A conspecific bioassay in L. riparia revealed that this factor is lipid-mobilizing. Isolation of the neuropeptide was achieved by single-step RP-HPLC. The primary structure of the earwig adipokinetic peptide was elucidated by automated Edman degradation in combination with matrix-assisted laser desorption/ionization mass spectrometry. It is a blocked octapeptide, pGlu-Val-Asn-Phe-Ser-Thr-Gly-Trp-NH2, previously denoted as Grb-AKH and first identified in Gryllus bimaculatus ( Gade & Rinehart 1987). The synthetic peptide co-chromatographed under various conditions with the native peptide and, in biological assays, resulted in lipid-mobilization in L. riparia when injected in low concentration.

Solitary male nymphs of the cockroach Diploptera punctata (Eschscholtz) (Blattaria: Blaberidae) took significantly longer to reach adulthood than males paired with either a male or female nymph or grouped with four other male nymphs since birth. When isolated throughout nymphal development, 15.8% of males passed through 3 stadia before adult eclosion, and the remainder went through 4 stadia. In contrast, 61.3% of paired males became adults in 3 stadia. Males need not, however, be isolated or paired for the entire nymphal period to express isolated or paired patterns of development. About 60% of males paired in just the first stadium or its initial 9 days became adults in 3 stadia, and only 20.4% of males isolated in the first stadium and the first 3 days of the second reached adulthood within 3 stadia. Although the first stadium was a critical period in which social condition determined the course of future development, analyses of covariance showed that isolated males gained less weight than paired ones, not only in the first stadium, but in the second as well. Moreover, the degree of growth of a male in the second stadium, measured as either weight gain or relative growth rate, did not depend on the male’s social condition in the first stadium, because isolated second-instar males grew less than paired ones, even when both sets of insects had been paired in the first stadium. Female nymphal development, unlike that of males, was not greatly affected by social factors.

Ecophysiological features, including survival and recovery from freezing and determination of the freezable water content, are reported for a cold-adapted cockroach Celatoblatta quinquemaculata Johns 1966 (Dictyoptera, Blattidae) inhabiting alpine communities at altitudes greater than 1300 m a.s.l. in mountains of Central Otago, New Zealand. Nymphs ranged from 15 to 51 mg live weight of which 67% was water. Cockroaches had a mean supercooling point temperature of −5.4 ± 0.1°C; with recovery from freezing close to this temperature being rapid, but no recovery was observed when frozen at −9 to −10°C. The duration of exposure to freezing conditions and the time allowed for recovery (24–96 h) both influenced individual recovery and subsequent survival. Comparison of supercooling point data and survival shows that this species possesses a few degrees of freeze tolerance, and individuals have been found frozen in the field when subzero temperatures occur. Differential scanning calorimetry showed ≈ 74% of body water froze during cooling and between 24 and 27% of total body water was osmotically inactive (unfreezable under the experimental conditions). Carbohydrates, other than glucose at 7.5μg/mg fresh weight, were in low concentrations in the body fluids, suggesting little cryoprotection. No thermal hysteresis from antifreeze protein activity was detected in haemolymph samples using calorimetric techniques. It is suggested that slow environmental cooling rates, together with high individual supercooling points, confer a small amount of freezing tolerance on this species enabling it to survive low winter temperatures. This has allowed it to colonize and maintain populations in alpine habitats > 1300 m a.s.1. in New Zealand.

. Under laboratory conditions the hissing-cockroach Gromphadorhina portentosa (Schaum) forms clusters, which appears to be an adaptive behaviour to help reduce water loss. Adults grouped together retain water nearly twice as effectively as isolated individuals. This ‘group effect’ complements the cockroach's large body size (small surface area to volume ratio) to lower the rate of water loss still further. Despite a modest 28% tolerance limit for weight loss during dehydration, adult females survive absolute drying conditions of 0% relative humidity without food and free water for at least a month, showing their impressive capacity for water retention. Rates of water loss of immature adults correlate with size, and no transition temperature is detected in females. To replenish water stores, cockroaches drank liquid water; there is no evidence for water gain by water vapour absorption. The profound impact exerted by the ‘group’ for water conservation suggests that members of this species live huddled together in nature, particularly during the long tropical dry season in order to conserve water, and this adds to previous evidence for the existence of a probable social structure.

In females of Diploptera punctata the corpora allata undergo a gradual increase in volume during most of the second nymphal stadium. In the first half of the stadium, steady growth of the glands results from a progressive increase in the size of constituent cells. Late in the stadium, cell size declines but the volume of the glands continues to rise due to an increase in cell number. Changes in cell size during the stadium displayed a distinct pattern in relation to Juvenile Hormone (JH) synthesis. Both cell size and activity increased during the first two-thirds of the stadium, peaked early in the last third of the stadium, and decreased before the moult. The rise in cell numbers late in the stadium corresponded to a wave of cellular mitosis and occurred after a steep decline in the rate of JH biosynthesis. Exposure of late second instars to fenoxycarb. a JH analogue, depressed mitosis significantly, suggesting autocrine regulation of cell proliferation in the corpora allata. Possible mechanisms modulating sequential cycles of growth and atrophy of cells and cell proliferation in these glands are discussed in relation to temporal patterns of JH and ecdysteroid titres in nymphs.

The innervation of the gut of the cockroach Leucophaea madera (F.) has been studied by means of wholemount immunocytochemistry with antisera raised against Leu-callatostatin, a cockroach allatostatin homologue identified from neuropeptide isolation and gene studies in the blowfly Calliphora vomitoria. Leu-callatostatin-imunoreactive neurones in the brain, with axon trajectories in the stomatogastric nervous system, innervate the foregut and midgut. Neurones in the last abdominal ganglion supply the hindgut and the midgut via the proctodeal nerve. In addition to a rich callatostatin-immunoreactive nerve supply, the midgut, including the midgut caeca, contain numerous callatostatin-immunoreactive endocrine cells. Physiological studies show that the spontaneous contractile activities of the foregut, but not the hindgut, are inhibited by callatostatin neuropeptides. Leu-callatostatin 3 was the most potent of the range of Leu-and Met-callatostatins tested, with a dose-dependent response between 10-13 and 10-7 M. This is similar to the results obtained with the previously identified myoinhibitory peptide of L. maderae, leucomyosuppressin. However, this peptide, with a different type of structure to the allatostatins, inhibits both foregut and hindgut motility equally. Experiments with a series of analogues of the Met-callatostatins showed that the free acid (as opposed to the carboxyamidated peptide) and N-terminally truncated peptides were inactive. These morphological and physiological results are thought to be representative of the, as yet unidentified, naturally occurring allatostatin homologues of L. maderae. This family of peptides should be added to the increasing list of insect gut myoinhibitory substances.

The concentrations of fat body and haemolymph vitellogenin and ovarian vitellin during the first gonadotropic cycle of the cockroach Blattella germanica (L.) (Dictyoptera, Blattellidae) have been studied. For these purposes, a polyclonal antibody against B. germanica vitellogenin and vitellin has been obtained, and an ELISA to quantify these proteins has been developed. Ovarian vitellin levels follow a pattern which parallels those of basal oocyte growth and Juvenile Hormone production by the corpora allata. This suggests that Juvenile Hormone regulates vitellogenin uptake into oocytes. Fat body and haemolymph vitellogenin levels give cyclic and parallel patterns. However, the cycle of Juvenile Hormone appears delayed with respect to that of vitellogenin. We suggest that the production of Juvenile Hormone, although cyclic in profile, does not modulate alone the cycle of vitellogenin. At least a supplementary mechanism, apparently independent of Juvenile Hormone, may be involved in the decline of vitellogenin production at the end of the vitellogenic cycle.

Larval holometabolous insects show changes in behaviour (e.g. start of wandering and spinning) in specific periods of the moulting cycle in relation to definite ratios of juvenile hormone and moulting hormone (ecdysone). In hemimetabolous insects no such changes in behaviour are known. It should be investigated whether the cockroach Periplaneta americana shows changes in locomotor activity and in food and water consumption in relation to periods of ecdysone production during the last larval stage. Within a mean duration of the last larval stage of 30 days there were two periods of reduced locomotor activity: on day 9 and between days 13 and 17. From days 12–13 food consumption decreased by c. 40% up to the day 18. Water consumption decreased between days 9 and 18 by about 55%. Peaks of ecdysone production appeared after these changes of behaviour in each case. Therefore in larval Periplaneta ecdysone seems not to trigger these behavioural changes.

Previous research has shown that fundamental properties of the circadian pacemaker that drives the rhythm of locomotor activity in the cockroach Leucophaea maderae L. are permanently altered by exposure of animals to 22 or 26 h light cycles during post-embryonic development (Barrett & Page, 1989; Page & Barrett, 1989). The present results document differences between animals exposed to either constant darkness (DD) or constant light (LL) during postembryonic development in the free-running period, the phase shifting response to light pulses, and the response to an LL to DD transition of the adult pacemaker. In addition, the changes in pacemaker period and in the phase shifting response that result from raising animals in several different lighting conditions are shown to be strongly correlated. The data suggest there is a developmentally labile interdependence between the period of the pacemaker and its sensitivity to light.

Food consumption and reproduction were compared through two gonotrophic cycles in female brown-banded cockroaches, Supella longipalpa (F.), fed a standard rat food diet which had been serially diluted with methyl cellulose, alpha cellulose or dextrin. Females did not increase daily intake to compensate for dilution, and reproductive rate was highly dependent upon the degree of dilution of the diet. With increasing dextrin concentrations, digestibility increased, but reproductive rate, size and mass of oothecae, and efficiency of food utilization progressively decreased. On diets containing 75% rat food and 25% methyl cellulose, females consumed minimal amounts of food and 80% of the females did not mate or produce oothecae within 30 days. On identical diets containing 25% alpha cellulose, food digestibility decreased and females ate significantly more than females fed 25% methyl cellulose, but less than females fed rat food. Many (65%) females fed 25% alpha cellulose produced oothecae which were smaller and took longer to form than in control females fed on rat food. At concentrations higher than 50% of either methyl or alpha cellulose daily intake was further reduced and females died rapidly.

The effects of age, time ofday and pheromone concentration the responsiveness of the male brown-banded cockroach, Supella longipalpa (F.), to female volatile pheromone examined. Male response increased with age and peaked 10 days after the imaginal moult. A diel periodicity of responsiveness observed, with greater responses in the scotophase. The behavioural response dose- dependent and its diel periodicity most obvious when intermediate pheromone concentrations (e.g. 0.01 female equivalents) used. Higher concentrations obscured the periodicity by eliciting greater responses in the photophase, while the lower concentrations did by decreasing the response in the scotophase. Following entrainment to LD 12:12h cycle, the periodicity of response to 0.01 female equivalents of pheromone persisted for at least 54 h under continuous darkness, further demonstrating true circadian rhythm.

The amount of the major component of the cuticular sex pheromone, 3, 11-dimethyl-2-nonacosanone, on individual female German cockroaches, Blattella germanica (L.), as a function of age was determined by gas-liquid-chromatographic analysis. Accumulation of phermone increased with age in both virgin and mated females. During the first gono-trophic cycle, the pheromone accumulated most rapidly when oocyte growth rates were maximal (days 5–10), and least rapidly while the female carried an ootheca (days 11–32). Pheromone accumulation was similar in virgin and mated females when the same physiological stages (determined by oocyte maturation) were considered. Inhibition of Juvenile Hormone release, through allatectomy, chemicals (precocene or fluoromevalonate), or through mechanical egg case implants, suppressed or delayed pheromone production and oocyte growth. The Juvenile Hormone analogue ZR512 induced allatectomized or head-ligated females and females with chemically or mechanically inhibited corpora allata to produce pheromone and enlarge their basal oocytes. Finally, ZR512 applied to intact females stimulated pheromone production in a dose-dependent manner.

Ovariectomy has been used to study the role of the ovary in endocrine homeostasis. Our studies on young virgin adults of the cockroach Blattella germanica (L.) (Dictyoptera, Blattellidae) show that the cytological development of the corpora allata (CA) in ovariectomized females proceeds as in intact specimens, whereas the rates of Juvenile Hormone (JH) synthesis are lower. Stimulation of the CA from ovariectomized females in vitro by mevalonolactone suggests that enzymatic mechanisms which follow mevalonate formation in the biosynthetic pathway are functional. The synthetic capabilities of these CA are also illustrated by the kinetics of JH production in vitro, because hormonal release increases with time to reach 'normal' levels after 8h of incubation. Our data suggest that the absence of ovaries leads to effective inhibition of JH biosynthesis rather than to an impairment of the developmental process in the CA cells.

Nymphs of Blaberus craniifer were exposed to a point source of light while walking on a servosphere apparatus. The initial response was a brief stop followed by running at a rate exceeding that in darkness. Their course was away from the light source and their direction became closer to 180° away from the light, as the light source approached the horizontal. The straightness of the course was controlled by both internal and external orientation information. If the source was overhead the course was more circuitous than with the directional light. Following a temporal switch from darkness to light, abrupt sharp turns of more than 90° led the animals back into a ‘zone of darkness’. Stopping occurs immediately after the onset or offset of the light stimulus, but the duration of stops does not seem to be correlated with the time since a previous stop, nor with the length of the run prior to a stop. Responses to a sudden onset of a light stimulus seem to represent adaptations for re-locating a ‘zone of darkness’ and for predator avoidance when a cockroach is disturbed during its photophase.

Extracts of corpora cardiaca from two cockroaches, Nauphoeta cinerea Olivier and Leucophaea maderae F., from a cricket, Gryllus bimaculatus De Geer, from the Colorado potato beetle, Leptinotarsa decemlineata Say, and from the sphinx moth, Sphinx ligustri L. were assayed for adipokinetic and hypertrehalosaemic activity, in acceptor locusts (Locusta migratoria L.) and cockroaches (Periplaneta americana L.) respectively. Both bioassays give positive results with all corpus cardiacum material tested except that from the sphinx moth; in this insect haemolymph lipid concentrations (but not those of the total carbohydrate) are, however, increased after injection of an extract of corpora cardiaca from the same species. A similar result is obtained when specimens of G. bimaculatus are injected with an extract of corpora cardiaca from G. bimaculatus. Biological activities of corpus cardiacum extracts from all species investigated can be resolved on reversed-phase high-performance liquid chromatography. Gland extracts from the two cockroach species each show a single absorbance peak which has hypertrehalosaemic activity, but with a (common) retention time distinct from all previously described arthropod neuropeptides. The corpora cardiaca of G. bimaculatus contain also a novel adipokinetic factor with a retention time distinct from previously characterized arthropod hormones, as well as from the new cockroach factor described in this study. The two hypertrehalosaemic factors from the corpora cardiaca of the potato beetle coelute with the hypertrehalosaemic hormones I and II of the American cockroach. The active (adipokinetic) compound from glands of S. ligustri appears to coelute with locust adipokinetic hormone I.

Two peptides, HGHI and HGHII, both inducing hyperglycaemia and activation of fat body glycogen phosphorylase can be isolated from the corpora cardiaca of the American cockroach, Periplaneta americana, using high-performance liquid chromatography. Both peptides are N-terminally blocked by a pyroglutamate residue and are thus not available for sequencing methods using the Edman degradation as this technique requires a free N-terminus. The blocked peptides were treated with pyroglutamate aminopeptidase to cleave the pyroglutamate residue, and the C-terminus of each peptide is also blocked and neither molecule can be cleaved by carboxypeptidase A. The following sequences for hyperglycaemic hormones HGHI and HGHII have been revealed using gas-phase sequencing. HGHI pGlu-Val-Asn-Phe-Ser-Pro-Asn-Trp-NH2 HGHII pGlu-Leu-Thr-Phe-Thr-Pro-Asn-Trp-NH2 Both peptides show remarkable similarities to locust adipokinetic hormones I and II and prawn red-pigment concentrating hormone, and are identical to two myotropic peptides MI and MII (O'Shea et al., 1984; Witten et al., 1984) and two cardioactive and hyperglycaemic peptides CC-1 and CC-2 (Scarborough et al., 1984), respectively, also isolated from the corpora cardiaca of P. americana.

Effects of early post-embryonic development and ageing on the circadian rhythm of locomotor activity in the cockroach, Leucophaea maderae, were investigated. Cockroach nymphs (first to fourth instars) were found to exhibit a circadian rhythm of activity generally similar to that of adults, but there appeared to be a significant change in pacemaker period (τ) early in nymphal development, and nymphs exhibited regular fluctuations in activity level which may be related to the moulting cycle. Mean τ of adults was remarkably stable with age – no significant change was found in either males or females throughout the life span of the adult. There was, however, a small but significant difference between the average period (τ) of adult males (τ= 23.72 ± 0.12h) and adult females (τ= 23.84 ± 0.13 h). Lighting conditions during post-embryonic development were found to have major effects on τ of adults. Males raised in constant darkness had a significantly shorter period (τ= 23.52 ± 0.11 h) than males raised in LD 12:12, and adult males and females which had been raised in non-24-h light cycles (T = 22 h or 26 h) exhibited major differences in τ from animals raised in LD 12:12 which persisted for several months. Animals exposed as adults to non-24-h light cycles also showed ‘after-effects’ on τ, but the magnitude of the effect was much less than that exhibited by animals exposed as nymphs.

Recognition of Periplaneta americana (L.) oothecae by the egg parasitoid Tetrastichus hagenowii (Ratzeburg) (Hymenoptera: Eulophidae) is mediated by mucopolysaccharides from the salivary glands, and calcium oxalate from the colleterial glands of the roach. Calcium oxalate, the most active compound isolated, only elicits stereotyped host recognition behaviour from the parasitoid when perceived in the context of a curved surface. Although empty oothecae contain calcium oxalate, female T. hagenowii appear to determine the unsuitability of these oothecae soon after drumming begins and abandon them. The potential usefulness of host recognition kairomones in biological control is discussed.

The corpora cardiaca of the stick insect, Sipyloidea sipylus Westwood, contain peptidic material which elevates blood lipids in migratory locusts, blood carbohydrates in American cockroaches, and activates glycogen phosphorylase in the fat body of the cockroach in a time- and dose-dependent manner. The active principle is found in appreciable amounts only in the corpora cardiaca; slight hyperlipaemia is caused by extracts made from corpora allata and abdominal ganglia, whereas brain, suboesophageal and thoracic ganglia are not active. The adipokinetic activity is already present in corpora cardiaca from second instar (first day) nymphs. The factor retains its adipokinetic activity after boiling for up to 1 h. Conspecific injections of extracts from corpora cardiaca of S.sipylus cause hypertrehalosaemia in ligated stick insects and activate glycogen phosphorylase in non-ligated S.sipylus. After incubation of corpora cardiaca in vitro in saline with high concentrations of potassium and calcium, one fraction with adipokinetic (in locusts) and hypertrehalosaemic (in stick insects) activity can be isolated from the medium by RP-HPLC. Fractionation of a methanolic extract of corpora cardiaca from S.sipylus by RP-HPLC shows that active compounds are confined to apparently three absorbance peaks. The material of the highest absorbance peak was purified to homogeneity by RP-HPLC, and its amino acid composition determined after acid hydrolysis with HCl and with methanesulfonic acid revealed the residues Asx, Thr(3), Glx, Pro, Gly, Leu, Phe and Trp. The primary structure of this hypertrehalosaemic factor is assigned as a blocked decapeptide, pGlu-Leu-Thr-Phe-Thr-Pro-Asn-Trp-Gly-Thr-NH2, from its FAB spectrum and metastable scans of its FAB spectrum. The structure is confirmed by synthesis; the synthetic and natural peptide co-chromatograph, and the synthetic peptide elevates blood carbohydrates in ligated stick insects and activates fat body phosphorylase in non-ligated S.sipylus.

The neurosecretory corpora cardiaca/corpora allata complex of Carausius morosus, a wingless insect of the superorder Orthopteroidea, contains adipokinetic and hyperglycaemic factor(s) capable of elevating lipids in locusts, and carbohydrates in cockroaches. Neither activity can be demonstrated in the stick insect itself, however. In addition, in locusts the Carausius gland extract is able to elevate levels of cyclic AMP in the fat body.

Some sublethal effects of tetrodotoxin (TTX) on the cockroach were investigated using chronically implanted extracellular electrodes. Prostration and subsequent paralysis were correlated with blockage of peripheral nerves, the CNS being apparently unaffected at the dose employed. Over several days following treatment many cockroaches showed a reversal of the symptoms and finally recovered fully. This coincided with recovery of the blocked nerves. It is suggested that TTX can be used to distinguish pharmacologically the peripheral and central actions of drugs and insecticides. Furthermore, TTX treatment could also be useful in the analysis of the subcellular nature of the sodium channels in nerve membrane.

The purpose of this study was to explore the olfactory recognition mechanisms used by individual subordinate male cockroaches, Nauphoeta cinerea (Olivier), in a dominance hierarchy. Results of two independent sets of experiments, one using an olfactometer and the other using an arena, suggest that olfactory cues are important to males. Subordinate males did not avoid dominant male odour in an olfactometer but could discriminate between the odours of familiar and unfamiliar individual dominant males, preferring the odour of the dominant male with which they had previous social contact. Since test subordinates did not avoid dominant male odours in the olfactometer, it is possible that the distance an individual remains from a dominant is important. To test this, individual dominant and subordinate males were tethered in the centre of an arena. Subordinate males were introduced into the arena and allowed to approach the tethered male. They approached tethered dominants and subordinates differently, and tended to keep away from the dominants. Tethered males unfamiliar to the test animals were avoided as well.

The ability of male cockroaches, Nauphoeta cinerea and Periplaneta americana, to respond directionally to tactile agonistic acts was tested using stimulation by artificially manipulated appendages. Responses by Nauphoeta included turning towards the stimulus at preferred angles of c. 40°, 90° and 180°, apparently relying on internally-stored directional sensory information. This turning responsiveness depended in part on the social status of the receiving individual, since subordinate individuals often retreated or failed to respond. Periplaneta males reacted to tactile stimuli by quick movements away from the stimulus or by kicking towards it. The leg used in kicking was that nearest to the part of the body which was stimulated.

Pure adipokinetic hormone (AKH) extracted from the glandular lobes of locusts induces hyperglycaemia in cockroaches. Larger doses (20–200 pmol) of AKH are required to induce hyperglycaemia in the cockroach than are required to produce hyperlipaemia in locusts (1–20 pmol).