Fifth-instar Larvae Research Articles

Decreased Response to Feeding Deterrents Following Prolonged Exposure in the Larvae of a Generalist Herbivore, Trichoplusia ni (Lepidoptera: Noctuidae)

We investigated the role of experience with several antifeedants on the feeding behavior of a generalist herbivore, Trichoplusia ni. Second-, third-, and fifth-instar larvae of T. ni were examined for their feeding responses to plant extracts (Melia volkensii, Origanum vulgare) and individual plant allelochemicals (cymarin, digitoxin, xanthotoxin, toosendanin, and thymol), after being exposed to them continually beginning as neonates. All tested instars of T. ni were capable of showing a decreased antifeedant response following prolonged exposure to most of the antifeedants tested compared with their naive conspecifics. Cardenolides (digitoxin and cymarin) were the exceptions. The response to oregano was affected as a result of previous exposure to different concentrations of oregano, unlike M. volkensii, leading us to conclude that T. ni sensitivity varies between stimuli and cannot be generalized. To demonstrate that decreased deterrence following prolonged exposure to M. volkensii was the result of learned habituation, three aversive stimuli were used. A high concentration of the particular antifeedant, xanthotoxin, acted as a noxious stimulus and dishabituated (reversed) the decrement in the antifeedant response to M. volkensii. Cold shock or CO2 was marginally effective in dishabituating the response. The fact that the decrease in antifeedant response can be dishabituated has implications for pest management.

Role of neurosecretory cells in the photoperiodic induction of pupal diapause of the tobacco hornworm Manduca sexta.

In the tobacco hornworm, Manduca sexta, pupal diapause can be induced by exposure of fifth-instar larvae to a short-day photoperiod. We studied the effect of surgical ablation of tissues containing the neurosecretory cells of the brain of fifth-instar larvae on the photoperiodic induction of pupal diapause. At the end of the experiments, we immunostained the neurosecretory cells to determine the success of the ablations. Under long-day conditions (LD 16:8 at 22 degrees C), all intact larvae, most of the sham-operated larvae, and control-operated larvae developed into nondiapausing pupae. Under short-day conditions (LD 10:14 at 22 degrees C), most intact, sham-operated, and control-operated larvae developed into diapausing pupae. Removal of type-II cells did not interfere with the photoperiodic response. Under long-day conditions, elimination of type-Ia(1) cells did not affect the incidence of nondiapausing pupae. When type-Ia(1) cells were removed under short-day conditions, however, the incidence of nondiapausing pupae was higher (51%, n = 41) than that of the intact (16%, n = 75), sham-operated (24%, n = 88), control-operated larvae (5%, n = 40), and larvae with type-II cells removed (11%, n = 27). Thus, removal of type-Ia(1) cells can impede induction of diapause. These results indicate that the type-Ia(1) neurosecretory cells have an important role in the induction of pupal diapause.

Feeding Behavior of Spruce Budworm (Lepidoptera: Tortricidae) Larvae Subjected to Multiple Exposures of <I>Bacillus thuringiensis</I> Variety <I>kurstaki</I>

Feeding behavior of the spruce budworm, Choristoneura fumiferana (Clemens), in multiple exposures to Foray 48B formulation of Bacillus thuringiensis Berliner subsp. kurstaki (Btk) was studied in the laboratory. A significant proportion of fourth- and fifth-instar larvae that were exposed to a Btk-contaminated food completely abstained from feeding on that food, even if these larvae had not previously been exposed to Btk-contaminated food. The occurrence of this behavior was higher in fourth-instar than in fifth-instar larvae and decreased with increasing fifth-instar weight. It is suggested that only larvae that have a reduced appetite can successfully abstain from feeding on the Btk-contaminated food over a 24-h period and that active ingredients of Btk or fermentation byproducts associated with the crystal/spore suspension are involved in the process. Spruce budworm did not exhibit aversion learning to Btk, which lead us to conclude that the field efficiency of multiple Btk applications should not be reduced by the behavioral response of spruce budworm larvae.

Lethal and sublethal effects of single and double applications of Bacillus thuringiensis variety kurstaki on spruce budworm (Lepidoptera: Tortricidae) larvae.

We conducted laboratory experiments to examine the effects of single versus double exposures of spruce budworm, Choristoneura fumiferana (Lepidoptera: Tortricidae) female larvae to various concentrations of a Bacillus thuringiensis variety kurstaki (Btk) commercial formulation (Foray 48B). Our main objective was to document the vulnerability to Btk and the sublethal responses of fifth-instar larvae that survived from a first ingestion of Btk during their fourth stadium and to compare them with insects treated either during their fifth or fourth stadium only. As reported in the literature, fifth-instar larvae were more vulnerable than fourth-instar larvae, but only at low and medium concentrations. Fifth-instar larvae that had survived Btk ingestion during their fourth stadium were more vulnerable to a high concentration of Btk and had a shorter feeding inhibition period than those that had not been exposed during their fourth stadium. Compared with a single treatment at the fourth stadium, a double exposure to Btk further reduced the population by 20-30%, depending on the concentration applied. The second treatment also induced another feeding inhibition period and increased larval development time by 14%. The impact of the different treatments on pupal weight depended on whether treated insects exhibited supernumerary instars. In the absence of developmental polymorphism, a higher concentration, a late, or a double exposure to Btk significantly reduced pupal weight.

Thermal death kinetics of fifth-instar Plodia interpunctella (Lepidoptera: Pyralidae).

Heat treatments have been suggested as alternatives to chemical fumigants for control of postharvest insects in dried fruits and nuts. Conventional forced hot air treatments heat product too slowly to be practical, but radio frequency treatments are capable of more rapid product heating. While developing radio frequency heat treatments for dried fruits and nuts, the heat tolerance of nondiapausing and diapausing fifth-instar larvae of the Indianmeal moth, Plodia interpunctella (Hübner), was determined using a heating block system developed by Washington State University. Both a 0.5th order kinetic model and a classical empirical model were used to estimate lethal exposure times for temperatures of 44-52 degrees C for nondiapausing fifth-instar larvae. We obtained 95% mortality at exposures suitable for practical radio frequency treatments (< or = 5 min) with temperatures of 50 and 52 degrees C. Diapausing larvae were significantly more tolerant than nondiapausing larvae at the lowest treatment temperature and shortest exposure, but differences were not significant at more extreme temperature-time combinations. Previous studies showed that fifth-instar larvae of the navel orangeworm, Amyelois transitella (Walker), were more heat tolerant than either diapausing or nondiapausing Indianmeal moth larvae. Consequently, efficacious treatments for navel orangeworm would also control Indianmeal moth.

Thermal Death Kinetics of Fifth-instar Plodia interpunctella (Lepidoptera: Pyralidae)

Heat treatments have been suggested as alternatives to chemical fumigants for control of postharvest insects in dried fruits and nuts. Conventional forced hot air treatments heat product too slowly to be practical, but radio frequency treatments are capable of more rapid product heating. While developing radio frequency heat treatments for dried fruits and nuts, the heat tolerance of nondiapausing and diapausing fifth-instar larvae of the Indianmeal moth, Plodia interpunctella (Hübner), was determined using a heating block system developed by Washington State University. Both a 0.5th order kinetic model and a classical empirical model were used to estimate lethal exposure times for temperatures of 44–52°C for nondiapausing fifth-instar larvae. We obtained 95% mortality at exposures suitable for practical radio frequency treatments (≤5 min) with temperatures of 50 and 52°C. Diapausing larvae were significantly more tolerant than nondiapausing larvae at the lowest treatment temperature and shortest exposure, but differences were not significant at more extreme temperature-time combinations. Previous studies showed that fifth-instar larvae of the navel orangeworm, Amyelois transitella (Walker), were more heat tolerant than either diapausing or nondiapausing Indianmeal moth larvae. Consequently, efficacious treatments for navel orangeworm would also control Indianmeal moth.

Does pheromone-based aggregation of codling moth larvae help procure future mates?

In field and laboratory bioassay experiments, we show that larvae of the codling moth, Cydia pomonella, cocoon in aggregations. This aggregation behavior of fifth-instar larvae prior to pupation and arrestment of eclosed adult males by mature female pupae seems to allow mating as soon as an adult female ecloses. This synchronous timing is realized because foraging fifth-instar are attracted by cocoon-spinning larvae and prepupae, but not by pupae, and because male pupae develop faster than female pupae. Eclosed males are arrested by sex pheromone that disseminates from female pupae even before adult females eclose. Communication in C. pomonella within and among developmental stages (larva-larva and pupa-adult, respectively) may be a strategy to procure mates. If so, our data add to current knowledge that attraction of mates in insects relies on communication among adults, or pupae and adults.

COMBINED EFFECT OF FOLIAR AND MYCORRHIZAL ENDOPHYTES ON AN INSECT HERBIVORE

Many foliar endophytic fungi deter herbivores from feeding on their host plants, but a previous study has suggested that their deterrent effect may be reduced in the presence of arbuscular mycorrhizae. In our study, factorial experiments determined the effects of infection of perennial ryegrass, Lolium perenne, by the foliar endophyte Neotyphodium lolii [present (+E) or absent (−E)] and the mycorrhizal fungus Glomus mosseae [present (+M) or absent (−M)] on larvae of the noctuid moth Phlogophora meticulosa. When host plants received adequate phosphorus (P), mycorrhizae and the foliar endophyte decreased the survivorship of larvae between their second and fifth instar in an additive manner. Sixth-instar larvae feeding on −M−E plants attained greater mass than those feeding on plants with one or both symbionts. There was an M × E interaction with the total amount of grass consumed per larva. The foliar endophyte increased relative consumption rate and reduced efficiency of conversion of ingested food of larvae, while the mycorrhizal fungus had no effect on insect nutritional indices. Larvae preferred −E over +E leaf blades regardless of mycorrhizal infection status. They showed no preference with respect to plant mycorrhizal status. When foraging among mixed stands of intact plants, larvae inflicted more damage on −E than on +E plants. Mycorrhizae reduced feeding damage, but only in −E plants and to a lesser extent than the foliar endophyte. The relative difference between +E and −E plants, in terms of grazing damage, was decreased but not eliminated by mycorrhizae. When P was limiting to plant growth, mycorrhizae still decreased the survivorship of larvae between their second and fifth instar, independently of the foliar endophyte. They accelerated larval development slightly. The foliar endophyte increased the survivorship of fifth-instar larvae, independently of mycorrhizae. We concluded that both foliar endophytes and mycorrhizae affected larval growth and survivorship. Some effects were additive, while others were nonadditive and depended to some extent on host P nutrition. Our findings suggest that the beneficial effect of the foliar endophyte on the host plant, in terms of insect resistance, was reduced but not eliminated by mycorrhizae.

BmNPV alters NADP-dependent malate dehydrogenase activity and associated macromolecules and retards growth and development of the mulberry silkworm,Bombyx moriL., during the final instar

Changes in tissue-specific NADP-dependent malate dehydrogenase (MDH) activity and protein and cholesterol contents and retardation of growth and development in the mulberry silkworm, Bombyx mori L., induced by infection with a baculovirus, the B. mori nucleopolyhedrovirus (BmNPV), were investigated. The study revealed that the relative growth rate and development of the fifth-instar larva was significantly inhibited during the progression of nuclear polyhedrosis disease at various times post inoculation (p.i.). Percent pupation and reproductive output were also adversely affected. NADP-dependent MDH activity in the hemolymph peaked sharply at 6 h p.i. and then gradually decreased, reaching a minimum at 264 h p.i., with exceptions at 30 and 72 h p.i. These levels of MDH activity were different from those in non-infected controls. In fat-body tissues, NADP-dependent MDH activity was significantly higher in infected insects than in non-infected controls of the same age. These results indicate that baculovirus infection causes significant changes in intermediary metabolic pathways, causing a significant fall and rise in protein and cholesterol contents in tissues during development of fifth-instar larvae.

Dynamic development of Trypanosoma cruzi in Rhodnius prolixus: role of decapitation and ecdysone therapy.

Decapitation and ecdysone therapy on the population dynamics of the Trypanosoma cruzi Dm28c clone in the stomach, small intestine and rectum of fifth-instar larvae of Rhodnius prolixus were investigated. Parasites were not found in the small intestine and rectum of decapitated insects after 10 days post-infection (p.i.). Decapitated ecdysone-supplemented insects sustained the flagellate infection in both gut compartments. In the rectum, the population density of parasites increased 5-fold in ecdysone-treated decapitated larvae and 7-fold in control insects. Epimastigote forms dominated with 40-65%, intermediate stages and round forms varied over 10-35% in the stomach, small intestine and rectum in both insect groups. Low numbers of metacyclic trypomastigotes were observed in the stomach and small intestine of the control group and decapitated insects supplemented with ecdysone but, at 15 days p.i., this form of flagellate reached about 20% in the rectum of the control insects. In the entire gut, at 30 days p.i., 23% of parasites in the control group and 8% in the decapitated insects treated with ecdysone were found. These results indicate that a head factor, possibly the prothoracicotropic hormone from the brain which stimulates ecdysone production by the prothoracic glands, may act directly or indirectly to stimulate the development of epimastigotes and round forms of the parasite and that a single ecdysone treatment is not able to fully reverse metacyclogenesis in decapitated R. prolixus.

Absorption and storage of phosphorus by larval Manduca sexta

The role of phosphorus (P) in numerous important biological structures, coupled with the observation that P-content of many insect foods is disproportionately low, suggests that P may be a critical nutrient for growing insects — however, the few studies examining the effects of dietary P on insect performance have generally found only weak relationships. This mismatch may be reconciled by understanding the physiological mechanisms by which insects handle P. Here we describe P processing by larvae of Manduca sexta. When given un-manipulated leaves of a common host plant, Datura wrightii, fifth-instar larvae retained about 85% of P consumed; when given P-enriched leaves larvae retained only 25% of P consumed. Analysis of gut concentrations of P at four sites along the digestive tract, and in leaves and feces, indicates that the rectum is the primary site of P transport between the gut and body and that differences in P retention may be accounted for by differential rates of rectal P transport. Larvae given P-enriched leaves also showed an eightfold increase in the concentration of P in the hemolymph, primarily as α-glycerophosphate — but only a 12% increase in the concentration of P in body tissues, suggesting that hemolymph plays a central role in storage and buffering of P.

Characterization of a novel silkworm (Bombyx mori) phenol UDP-glucosyltransferase.

Sugar conjugation is a major pathway for the inactivation and excretion of both endogenous and exogenous compounds. We report here the molecular cloning and functional characterization of a phenol UDP-glucosyltransferase (UGT) from the silkworm, Bombyx mori, which was named BmUGT1. The complete cDNA clone is 1.6 kb, and the gene is expressed in several tissues of fifth-instar larvae, including fat body, midgut, integument, testis, silk gland and haemocytes. The predicted protein comprises 520 amino acids and has approximately 30% overall amino-acid identity with other members of the UGT family. The most conserved region of the protein is the C-terminal half, which has been implicated in binding the UDP-sugar. BmUGT1 was expressed in insect cells using the baculovirus expression system, and a range of compounds belonging to diverse chemical groups were assessed as potential substrates for the enzyme. The expressed enzyme had a wide substrate specificity, showing activity with flavonoids, coumarins, terpenoids and simple phenols. These results support a role for the enzyme in detoxication processes, such as minimizing the harmful effects of ingested plant allelochemicals. This work represents the first instance where an insect ugt gene has been associated with a specific enzyme activity.

Effects of cetyltriethylammonium bromide on the replication of Bombyx mori nucleopolyhedrovirus.

An experimental study was undertaken to quantify the effects of cetyltriethylammonium bromide (CTAB) on the replication of Bombyx mori nucleopolyhedrovirus (BmNPV) and the transcriptionalactivity of BmNPV ie-1 promoter. The results demonstrated that the budded virus (BV) titer rose about 3.7-fold by adding CTAB to the culture media up to 0.1 mu g ml(-1) in infected Bm-N cells with a wild-type BmNPV. The transient expression level of luciferase driven by BmNPV ie-1 promoter was enhanced by more than 3-fold in the presence of 0.1 mu g ml(-1) of CTAB in uninfected insect cells via a transient expression system. Contrary to the rise in BV titer, the polyhedra inside the nucleus of infected cells dropped linearly from 4.0 x 10(6) ml(-1) down to 2.1 x 10(6) ml(-1) with in a range of CTAB concentrations from 0 to 0.25 mu g ml(-1). The same trend in expression level of beta -galactosidase or phytase was given when the Bm-N cells or fifth-instar silkworm larvae infected with a recombinant BmNPV containing the beta -galactosidase or phytase reporter gene driven by the polyhedrin promoter. We deduced that CTAB appeared to affect the virus bi-phasic life cycle stages and production pathways, resulting in an enhancement in BV production and a suppression of occluded virus (OV) production and expression of foreign genes controlled by the polyhedrin promoter.

Primary and continuous midgut cell cultures from Pseudaletia unipuncta (Lepidoptera: Noctuidae).

Midgut epithelial cells were isolated from fifth-instar Pseudaletia unipuncta larvae by collagenase treatment of midgut tissue, and cultured in TNM-FH medium. Long-term continuous culture and maintenance of midgut cells were achieved with P. unipuncta armyworm intestinal cells. Several cells lines were obtained from these P. unipuncta primary cultures, and they have been subcultured and maintained for over 24 mo. The three major midgut cell types were present in the cultures, including stem (regenerative), columnar, and goblet cells. In vitro morphogenesis and differentiation of columnar and goblet cells from stem cells were observed. There appeared to be a cycle of cell death of goblet and columnar cells followed by their replacement from stem cells every 7-8 wk. After approximately six passages, the cell density in T-flasks appeared to be somewhat constant, reaching 10(3)-10(4) cells per milliliter of medium. The columnar cells are round to rectangular in shape and possess a brush border, while the goblet cells have a classic flask-like shape with a central cavity. Peritrophic membrane-like secretions were observed in all the culture flasks. Infection of these cells with multiply embedded nucleopolyhedrovirus was confirmed, and we conclude that these midgut cells can be used as an in vitro model system to study early events in baculovirus infection.

Burchellin: effects on Triatoma infestans and on Trypanosoma cruzi within this vector.

Supplementation of blood with the neolignan burchellin (50 microg/ml), a compound from the arboreous Lauraceae Aniba burchelli, affected the ingestion of blood and the course of excretion of fourth- and fifth-instar larvae of Triatoma infestans, the latter especially within the first 4 h after feeding. The total resultant weight loss of treated fourth instars within 24 and 48 h after feeding was only 24% and 28% vs 41% and 48%, respectively, in untreated bugs. In fifth instars, the total weight losses of untreated bugs within 24 and 48 h after feeding were 38% and 41% whereas the weight of treated bugs decreased by 28% and 34%, respectively. In a treatment of Trypanosoma cruzi-infected fourth instars, burchellin significantly reduced the population density of the established infection in the rectum at 5 and 10 days after feeding. This was especially due to a significant increase in the number of the main dividing stage, the epimastigote.

Lifetime Reproductive Success in the Solitary Endoparasitoid, Venturia canescens

Parasitoid wasps have long been considered excellent organisms in studies examining the evolution of reproductive and life-history strategies. In examining the lifetime reproductive success of parasitoids in the laboratory, most investigations have provided the insects with excess hosts and food, where they exist in a relatively constraint-free environment. Importantly, these conditions may not accurately reflect the true heterogeneity of natural systems, where suitable hosts and food sources are likely to be limiting. This study examines the influence of differences in host and food availability on reproductive and life-history parameters in an asexual strain of the solitary endoparasitoid, Venturia canescens (Hymenoptera: Ichneumonidae). Lifetime reproductive success in V. canescens was measured in response to temporal variations in host and food (honey solution) access. Cohorts of parasitoids were provided with 200 fifth-instar larvae of the Indian meal moth, Plodia interpunctella (Lepidoptera: Pyralidae), and food for variable periods daily after eclosion. V. canescens is synovigenic, and host-deprived wasps continued to mature eggs over the first few days after eclosion until the egg storage capacity was reached in the oviducts. When these wasps were subsequently provided with hosts, oogenesis resumed and continued until later in adult life. Constantly fed wasps lived longer and produced more progeny than wasps from cohorts which were alternately fed and starved or were starved from eclosion. Moreover, wasps with constant host and food access produced most progeny early in life and usually experienced prolonged periods of postreproductive survival. In contrast, the reproductive period of wasps with limited host access was more evenly distributed throughout the adult life. Consequently, the cumulative progeny production by V. canescens with constant food access was fairly uniform irrespective of host availability. Longevity and fecundity in V. canescens were positively correlated with adult size. However, variable host access had little effect on the longevity of wasps which were constantly supplied with honey. Over the first 2 days of adult life, variation in food access also had no effect on progeny production by V. canescens. We argue that manipulating temporal host and food access to parasitoids in the laboratory more closely approximates natural conditions, where these resources are likely to be spatially separated. Moreover, our findings suggest that many highly synovigenic parasitoids like V. canescens, which produce microtype (=hydropic) eggs, have a considerably higher reproductive potential than ovary dissections have revealed. Our findings are discussed in relation to life-history evolution in the parasitic Hymenoptera.

Developmental changes in the patterns of feeding in fourth- and fifth-instar Helicoverpa armigera caterpillars

Summary Data are presented for developmental changes in feeding behaviour within and across the fourth and fifth stadium of Helicoverpa armigera (Lepidoptera, Noctuidae) caterpillars fed nutritionally homogeneous semi-synthetic foods. We recorded the microstructure of feeding over continuous 12-h periods on consecutive days throughout the two stadia, and in one experiment recorded continuously for 21 h. Larvae in the two stadia showed the same general pattern of macro-events in feeding, including a similar duration of post-ecdysis fast, which was usually broken by consumption of the exuviae, and then a sustained period in which discrete meals on the experimental food were taken regularly. There were, however, some distinct differences in the patterns of meal-taking both between stadia and across different one-third time segments within stadia. Considering between-stadium differences, the proportion of time spent feeding differed significantly only in the last segment of the feeding period of the two stadia, with the value for the fourth-instar larvae being substantially greater than for fifth-instar larvae. As regards within stadium changes, the proportion of time feeding increased from the first to the second segment of both stadia. However, whereas the proportion of time feeding increased from the second to the final segment of the fourth stadium, it decreased across the same period in the fifth stadium. These patterns of changes in the proportion of time feeding within and between stadia, and their behavioural mechanisms (combination of meal durations and meal frequencies), can be explained only partially with reference to increasing food requirements with development. Three areas are identified where further study might help elucidate the reasons for the observed developmental changes in the microstructure of feeding: allometric constraint, the dynamic links between ingestion and post-ingestive processing, and ecological factors such as predation.

Evaluation of insecticides for the control of Helicoverpa armigera (Hübner) and Helicoverpa punctigera (Wallengren) (Lepidoptera: Noctuidae) on soybean, and the implications for field adoption

AbstractSeven insecticides were evaluated against first‐, third‐ and fifth‐instar Helicoverpa armigera (Hübner) and Helicoverpa punctigera (Wallengren) larvae using a leaf‐disc bioassay of field‐treated soybean. Significant differences were detected in age‐specific mortality, time to effect (LT50) and field persistence of the insecticides tested. With the exception of trichlorfon, the mortality of first‐instar larvae was similar in the different treatments to the commercial standard, methomyl. The mortality of third‐ and fifth‐instar larvae in the spinosad and chlorpyrifos treatments was also similar to that in the methomyl treatment, while the age‐specific mortality of the insecticides based on Bacillus thuringiensis and nucleopolyhedrovirus declined more rapidly, resulting in low mortality to fifth‐instar larvae. The LT50 of the biological‐based products were significantly greater than those of the conventional products. Only chlorpyrifos showed significant activity 1 week after treatment. The results suggest the need for a user education program to highlight the likely differences in the field performance of the newer biological‐based products compared with the conventional chemicals with which growers are familiar.

Octopamine mimics the effects of parasitism on the foregut of the tobacco hornworm Manduca sexta.

The parasitic braconid wasp Cotesia congregata lays its eggs inside the body of the larval stage of its host, the moth Manduca sexta. The Cotesia congregata larvae develop within the hemocoel of their host until their third instar, when they emerge and spin cocoons and pupate on the outer surface of the caterpillar. From this time until their death approximately 2 weeks later, the Manduca sexta larvae show striking behavioral changes that include dramatic declines in spontaneous activity and in the time spent feeding. Coincident with these behavioral changes, it is known that octopamine titers in the hemolymph of the host become elevated by approximately 6.5-fold. Octopamine is an important modulator of neural function and behavior in insects, so we examined hosts for neural correlates to the behavioral changes that occur at parasite emergence. We found that, in addition to the changes reported earlier, after parasite emergence (post-emergence), Manduca sexta larvae also showed marked deficits in their ability to ingest food because of a disruption in the function of the frontal ganglion that results in a significant slowing or the absence of peristaltic activity in the foregut. This effect could be produced in unparasitized fifth-instar larvae by application of blood from post-emergence parasitized larvae or of 10(-6)mol l(-1)d,l-octopamine (approximately the level in the hemolymph of post-emergence larvae). In contrast, blood from parasitized larvae before their parasites emerge or from unparasitized fifth-instar larvae typically had no effect on foregut activity. The effects of either post-emergence parasitized blood or 10(-6)mol l(-1) octopamine could be blocked by the octopamine antagonists phentolamine (at 10(-5)mol l(-1)) or mianserin (at 10(-7)mol l(-1)).

Neuroendocrine regulation of seasonal morph development in a bivoltine race (Daizo) of the silkmoth, Bombyx mori L.

We investigated the neuroendocrine regulation of the development of seasonal morphs in a bivoltine race (Daizo) of the silkmoth, Bombyx mori, by decerebration, the transplantation of brain–suboesophageal ganglion (Br-SG) complexes and the injection of active neuropeptides. When brains were removed from fresh pupae destined to develop into summer morphs (SD pupae) by embryonic and larval exposures to short days at low temperature, the pupae developed into autumn or intermediate morphs. However, in pupae destined to develop into autumn morphs (LD pupae), the operation did not show an effect on seasonal morph development. Br-SG complexes were excised from fifth-instar LD and fifth-instar SD larvae 2 days after larval ecdysis and were transplanted into the abdomen of SD larvae of the same age. The Br-SG complexes of LD larvae, but not the Br-SG complexes of SD larvae, shifted the host's seasonal morph development toward the autumn morph. Furthermore, when treated with crude pupal SGs extract and diapause hormone (DH), fresh SD pupae developed into autumn or intermediate morphs, respectively. Possibly the development of seasonal morphs in the silkmoth, B. mori, is regulated by a novel function of DH. Alternatively, DH may act on the imaginal wing disks at an earlier stage than on the ovaries.