Effects of juvenile hormone I and precocene I & II on Microplitis rufiventris when administered via its host, Spodoptera littoralis

We investigated the effect of juvenile hormoneI (JHI) and precocene I & II (PI & PII) onthe endo-developmental stages, reproductiveperformance and longevity of Microplitisrufiventris Kok. after topical treatment ofhost larvae of Spodoptera littoralis(Boisd.). Host larvae were treated with each ofthe test compounds by a single dose 2–4 h afterparasitization or 1 day prior to parasitoidemergence, or by repeated daily applications 2–4 h after parasitization to parasitoid emergence.JHI-treatments increased parasitoiddevelopmental time, induced developmentalabnormalities and inhibition of parasitoidegression. Interestingly, one of the causal factorsthat inhibited the parasitoid egression was themoulting of some parasitoid larvae into asupernumerary instar. Applications of PI or PIIto parasitized S. littoralis larvae didnot affect parasitoid developmental time, butsome developmental abnormalities were noted.Melanin formation in host's blood was greaterin PI-treatments than in PII-treatments.PII-treatments arrested development of somefirst instar parasitoid larvae typical of JHIexcess. Adult female wasps that emerged fromJHI-treated hosts lived for significantlylonger periods than control females, but didnot produce more offspring. The longevity andreproductive capacity of females whichdeveloped in hosts treated by precocenes werereduced compared with the females emerged fromcontrol hosts. Even the small number ofadult wasps from PII-treated hosts which livedfor the same period as control wasps, showed asignificant reduction in their reproductiveperformance. Repeated daily applications of thetest compounds reduced wasp fecundity andlongevity to a significantly greater degreethan the single applications, except for a lowdose of JHI (1 µg). The study demonstratesthat growth disrupters, such as JHs andprecocenes have a delayed developmental effect onM. rufiventris and points out the need tobe aware of these effects in implementingcontrol strategies of S. littoralis innatural habitats.     

A possible therapy by juvenile hormone I to Spodoptera littoralis (Boisd.) larvae previously treated by precocene II

An effective constant dose (55 μg) of precocene II (PII) was topically tested against the last three instars of Spodoptera littoralis (Boisd.) larvae. Application of PII induced morphogenetic abnormalities typical of juvenile hormone (JH) excess. The resultant imperfect insects included larval‐pupal mosaic and partial or severe cases of untanned pupae. The sixth‐instar larvae were more sensitive to PII administration than the two preceding instars. However, sensitivity of the last larval instar to PII varied with the timing of dose application relative to the developmental status of the larvae. Whereas the newly ecdysed (0‐day old) larvae were more sensitive, the older larvae of the same sixth‐instar showed sharp decrease in their sensitivity to PII with a concomitant increase of their age. Application of a single dose (5 μg) of JH I to PII pre‐treated larvae significantly (P < 0.001) reduced the production of imperfect insects where many PH‐treated larvae developed successfully to apparent normal pupae. Although a single dose of PII was more effective on S. littoralis larvae than repeated daily doses, the effectiveness of JHI‐therapy to PH pre‐treated larvae by repeated doses was less effective in producing perfect insects than JH‐therapy to PII pre‐treated larvae by single doses. The reversal of any of these by applied JHI is quite interesting but the mechanisms remain to be unraveled.     

Untersuchungen zur Veränderung der Unkrautflora bei mehrmaligem Getreidenachbau auf einer Tonschwarzerde des Thüringer Beckens

The latent effects of precocenes I and II (PI and PH) and juvenile hormone I (JHI) when topically applied to the last three instars of Spodoptera littoralis (Boisd.) larvae have been studied. Application of both PI or PII resulted in morphogenetic abnormalities resemble some effects induced by administration of JHI, e.g., larval‐pupal intermediate, partial or severe cases of untanned pupae and imperfect moths. In PII‐treatments, the effect was instar‐dose‐dependent. The intermediate dose (55 μg) was more effective on S. littoralis larvae than other doses. The effectiveness of both doses of 40 or 70 μg in production of deformed larvae and pupae decreased when applied as repeated doses instead of single ones. In Pi‐treatments, the lower dose (25 μg) was more harmful to Spodoptera larvae than the higher dose (70 μg). Repeated application by either lower or higher doses did not enhance the production of imperfect insects. Application of JHI induced symptoms ranging from supernumerary instars, larval‐pupal intermediate, untanned pupae and deformed adults. The effect was dose‐dependent. In all tested compounds, there apparently was a latent or delayed effects. Although the insects were treated while they were larvae the complete effects were not apparent until after the insect had become a pupa or an adult. More efforts will be needed to understand how does precocene interferes with the process of tanning or sclerotization?     

Developmental status of endo-developmental stages of Microplitis rufiventris kok. in Spodoptera littoralis (boisd.) larvae topically treated by juvenile hormones I and II

The juvenile hormones I and II (JHI and JHII) were topically applied at concentrations of 1 or 5 μg to Spodoptera littoralis larvae containing newly deposited eggs of Microplitis rufiventris. The study demonstrated that both hormones do influence several aspects of the development of the parasitoid after treatment of its host larvae. Of these effects: (1) significant lengthening of periods of eggs and larvae, (2) moulting failure of parasitic larvae, (3) moulting the parasitic larvae into supernumerary instar, (4) arrest of postembryonic development in the first larval instar, (5) reduction in the emergence rate of parasitoid larvae, and (6) influence on embryogenesis. The application of JHI was more effective in disrupting the endo-development of the parasitoid than JHII. In all tests, control hosts produced significantly (P < 0.01) more parasitoids than treated ones. Off-target effects on natural enemies may seriously limit the use of JHs, especially in integrated control programmes.     

Influence of Precocene II on the estimated changes within the haemocyte population of parasitized spodoptera littoralis larvae by microplitis rufiventris

Series of experiments were undertaken to investigate whether application of Precocene II (PII) to parasitized S. littoralis larvae can influence their THCs and DHCs. PII was applied in single dose treatments (70μg/5 μl/dose). Haemolymph samples were taken at daily intervals from day 3 to day 8 post‐treatments. Two distinct alterations were noted; first, PII‐treatments caused significant decrease in THCs. Once the parasitoid larvae emerge from either treated or untreated hosts, a sharp decline in THCs was observed. The second effect was changes in DHCs of S. littoralis larvae. In controls, haemocyte types could be arranged in descending order due to their relative occurrence into plasmatocytes (PLs), prohaemocytes (PRs), spherule cells (SPs), granular cells (GRs) and oenocytoids (OEs). This order changed in treated hosts to PLs, GRs, PRs, SPs and OEs, i.e., GRs that were less prevalent in the haemolymph of control larvae increased significantly in PII‐treated hosts. In the latters, sharp rise in GRs levels followed parasitoid emergence. OEs cells were scarce in both treated and untreated hosts. No significant difference was observed between the overall means of PLs in both types of host larvae. But GRs‐PRs was markedly altered in treated larvae.     

Effects of juvenile hormones and precocenes on the immune response of Spodoptera littoralis larvae to supernumerary larvae of the solitary parasitoid,Microplitis rufiventris Kok

The study represents one of the first direct tests of juvenile hormones (JHs) on the cellular immune response of a lepidopteran insect, Spodoptera littoralis (Boisd.), superparasitized by a solitary parasitoid, Microplitis rufiventris Kok. JHII-treated S. littoralis larvae exhibited a significant reduction in encapsulation response to the supernumerary M. rufiventris larvae at different developmental ages (0-4, 10-14 and 24-26 h) of supernumerary M. rufiventris embryos within 48 or 144 h after parasitization than either JHI-treated or control hosts. The reduction in encapsulation response was significantly greater in hosts which received higher JHII dose (5 &mgr;g) than those treated with lower one (1 &mgr;g).The progression of the encapsulation, culminating in melanization in the capsules occurred more frequently in precocene-treated hosts than either JH-treatment or control ones. Application of the anti-juvenile hormones (PI or PII) caused stronger encapsulation reaction to the injured supernumerary parasitoid larvae than JHI or JHII. In PI-treatments, the reaction was rapid and dramatic by lower (25 &mgr;g) dose treatments than higher ones (70 &mgr;g).The capsules formed around the supernumerary larvae of M. rufiventris showed signs of melanization within 48 h in PI-treated hosts. Melanization was more pronounced in either PI- or PII-treated hosts prior emergence of successful parasitoid larvae. Generally, the injured supernumerary larvae were seen within melanin-rich capsules in PI- and PII-treated hosts. Melanization was not observed in most capsules of JHI- and JHII-treated hosts. Additional physiological effects were observed: (1) in some cases, application of JHI to parasitized S. littoralis larvae inhibited subsequent emergence of its wasp, M. rufiventris, and (2) teratocytes (cells of wasp origin) from PI or PII-treated hosts were smaller in size than those observed in the JH-treated or control hosts.Finally, the dramatic increase in encapsulation responses of precocene-treated S. littoralis larvae of the supernumerary of M. rufiventris and definitely opposite reactions in typical hosts but JHII-treated hosts suggested that the cellular defense reaction may be under inhibitory hormonal control.The results of the study have implications for medically important insects such as mosquitoes and their interactions with parasites, as well as having significance for biocontrol and mass rearing programs using parasitoids.     

Diamondback moth females oviposit more on plants infested by non‐parasitised than by parasitised conspecifics

Abstract 1. When offered a choice, female diamondback moths (Plutella xylostella) oviposited more eggs on plants with non‐parasitised conspecific larvae than on plants with parasitised larvae. 2. The leaf area consumed by parasitised larvae was significantly lower than that by non‐parasitised larvae. However, this quantitative difference in larval damage did not explain the female’s ability to discriminate between plants with parasitised and non‐parasitised larvae, as females showed an equal oviposition preference for plants infested by higher or lower densities of non‐parasitised larvae. 3. Pupal weight and duration of the larval stage of P. xylostella were independent of whether larvae were reared on plants that were previously infested by either non‐parasitised or parasitised larvae. 4. The larval parasitoid Cotesia vestalis did not distinguish between plants infested by non‐parasitised larvae and plants infested by larvae that had already been parasitised by conspecific wasps. 5. Based on these data, it can be concluded that the moth oviposition preference for plants infested by non‐parasitised conspecifics relative to plants infested by parasitised conspecifics was not explained by plant quality or by the attractiveness of plants towards wasps. It is hypothesised that one of the reasons for this preference is avoidance of plants where a relatively high risk of parasitism is expected due to the emergence of parasitoids from the parasitised host larvae.     

Feeding of fall armyworm,Spodoptera frugiperda,on Bt transgenic cotton and its isoline

Studies on insect food intake and utilization are important for determining the degree of insect/plant association and host species’ resistance, and also for helping design pest management programs by providing estimates of potential economic losses, techniques for mass breeding of insects, and identifying physiological differences between species. We studied the feeding and development of fall armyworm, Spodoptera frugiperda (JE Smith) (Lepidoptera: Noctuidae), on transgenic (Bt) and non‐transgenic (non‐Bt) cotton. The larvae of S. frugiperda fed on Bt cotton had a longer development period (23.0 days) than those fed on non‐Bt cotton (20.2 days). Survivorship of S. frugiperda larvae fed on Bt cotton (74.1%) was lower than that of larvae fed on non‐Bt cotton (96.7%). Pupal weight of larvae fed on Bt cotton (0.042 g) was lower than that of larvae fed on non‐Bt cotton (0.061 g). The cotton cultivar significantly affected food intake, feces production, metabolization, and food assimilation by S. frugiperda larvae. However, it did not affect their weight gain. Intake of Bt‐cotton leaf (0.53 g dry weight) per S. frugiperda larva was lower than the intake of non‐Bt‐cotton leaf (0.61 g dry weight). Larvae fed on Bt‐cotton leaves produced less feces (0.25 g dry weight) than those fed on non‐Bt‐cotton leaves (0.37 g dry weight). Weight gain per S. frugiperda larva fed on Bt‐cotton leaves (0.058 g dry weight) was similar to the weight gain for larvae fed on non‐Bt‐cotton leaves (0.056 g dry weight). The cotton cultivar significantly affected the relative growth, consumption, and metabolic rates, as well as other nutritional indices: the figures were lower for larvae fed on Bt‐cotton leaves than for larvae fed on non‐transgenic cotton leaves.     

Phenotypically plastic traits regulate caste formation and soldier function in polyembryonic wasps

Polyembryonic encyrtid wasps are parasitoids that have evolved a clonal form of embryogenesis and a caste system where some progeny become reproducing wasps whereas others develop into a sterile soldier caste. Theory based on the biology of Copidosoma floridanum predicts that the primary role of soldier larvae is to mediate conflict over sex ratio, which also favours female‐biased soldier production. Other data, however, suggest that female‐biased soldier production reflects a developmental constraint. Here, we assessed whether female‐biased soldier function by polyembryonic wasps reflects sex‐specific adaptation or constraint by conducting comparative studies with Copidosoma bakeri, a species that produces clutch sizes similar to C. floridanum yet rarely produces broods associated with sex ratio conflict. Our results indicate that the oviposition behaviour of adults, development of progeny and function of soldier larvae differ greatly between C. bakeri and C. floridanum. These findings indicate that caste formation and soldier function in polyembryonic encyrtid wasps are regulated by phenotypically plastic traits. Our results further suggest that the primary function of the soldier caste in some species is defence of host resources from competitors whereas in others it is the resolution of sex ratio conflict.     

The α-amylase inhibitor acarbose does not affect the parasitoid Venturia canescens when incorporated into the diet of its host Ephestia kuehniella

Amylase inhibitors (AIs) are suitable candidates for protecting plants and their products from attacks by herbivorous and granivorous insects. However, detailed studies of the suppressive effects of AIs on target and non‐target insects are necessary before their application in post‐harvest protection. To address this issue, laboratory bioassays were used to test the effect of the non‐proteinaceous inhibitor acarbose on a stored product pest, the flour moth Ephestia kuehniella (Zeller) (Lepidoptera: Pyralidae), and its parasitoid Venturia canescens (Gravenhorst) (Hymenoptera: Ichneumonidae). Two sublethal concentrations (0.001 and 0.0001%, wt/wt) of acarbose were incorporated into the diet of parasitized and unparasitized larvae of E. kuehniella. Development time and fresh body weight of the larvae, together with the size of the wasps, were compared for insects reared on acarbose‐treated and control diets. On the diet containing 0.001% acarbose, the developmental time was longer and relative weight gains of the E. kuehniella larvae were lower, but the weight of the larvae prior to pupation was similar to that of the control. The acarbose did not have a suppressive effect on the parasitoid V. canescens; in fact the wasps that emerged from the hosts reared on a diet containing 0.0001% acarbose were on average larger and heavier than the controls. These results demonstrate that it might be possible to enhance the control of stored product pests by using both biological control and AIs.     

Age-dependent responses of last instar larvae of Spodoptera mauritia to precocene II

ABSTRACT. Last instar larvae of Spodoptera mauritia Boisd. (Lepidoptera: Noctuidae) of various ages were treated with a single dose of 80 μg or 160 μg of precocene II (PII) and the effects of larval—pupal transformation were studied. Final instar (sixth) larvae treated on day 0, 1 or 2 with 80 μg P II showed a high mortality; P II treatment during subsequent days of this stadium showed a diminishing tendency for mortality. P II prolonged the larval—pupal period in all larvae but the effect was somewhat reduced in the mid-stadium.
Treatments of 160μg P II to 0-day-old, 1-day-old, 2-day-old and 3-day-old larvae were highly toxic. Treatments of 4-day-old and 5-day-old larvae with 160 μg P II delayed pupation by 1 day, but in addition the larvae often moulted into larval—pupal intermediates. Co-application of 50μg juvenile hormone analogue (ZR 512) with 160 μg P II restored both normal morphogenesis and a normally-timed moult.     

Bio‐safety evaluation of Cry1Ac,Cry2Ab,Cry1Ca,Cry1F and Vip3Aa on Harmonia axyridis larvae

Dietary exposure studies are initial steps in environmental risk assessments of genetically engineered plants on non‐target organisms. These studies are conducted in the laboratory where surrogate species are exposed to purified and biologically active insecticidal compounds at higher concentrations than those expected to occur in transgenic crops foliage. Thus, dietary exposure (early tier) tests provide robust data needed to make general conclusions about the susceptibility of the surrogate species to the test substance. For this, we developed suitable artificial diet and used it to establish a dietary exposure test for assessing the toxicity of midgut‐active insecticidal compounds to the larvae of the Asian ladybird beetle Harmonia axyridis (Pallas) (Coleoptera: Coccinellidae). Using boric acid as a model compound, we validated the bioassay established for H. axyridis larvae. An artificial diet containing boric acid which negatively affected survival, development and adult weights was offered to larvae and indicated that the bioassay was able to detect toxic effects of insecticidal substances incorporated in diets. Using this dietary exposure test, environmental risk assessment of Cry1Ac, Cry2Ab, Cry1Ca, Cry1F and the non‐Cry protein Vip3Aa was evaluated by analysing pupation rates, adult emergence rates, 7‐day larval weights, and freshly emerged male and female weights among the toxin treatments and a pure artificial diet. These life‐table parameters did not vary among artificial diets containing 200 μg/g Bt proteins or pure artificial diet. In contrast, boric acid adversely affected all life‐table parameters. Thus on these bases, we concluded H. axyridis larvae are not sensitive to these Bt proteins expressed in genetically engineered crops.     

Body size affects host defensive behavior and progeny fitness in a parasitoid wasp,Lysiphlebus fabarum

This study tested effects of maternal body size on foraging behavior and progeny development in a thelytokous population of Lysiphlebus fabarum (Marshall) (Hymenoptera: Braconidae). Small and large wasps were reared from first and second instar hosts [black bean aphid, Aphis fabae Scopoli (Hemiptera: Aphididae)], respectively, and each was provided with a patch (bean leaf disk) containing either 15 small (second instar) or 15 large (fourth instar) hosts for a 30‐min foraging period. Neither body size nor host size affected time allocation to various behaviors within a patch, but second instar aphids produced significantly more mummies than fourth instars. The preferred attack orientation was from the side of the aphid, suggesting wasps were sensitive to the risk of smearing with cornicle secretions. Few wasps developed in fourth instar hosts, suggesting later host instars were somewhat resistant to parasitism. Second instar hosts, the most suitable stage for L. fabarum development, relied more on defensive behavior, specifically kicking and secreting cornicle droplets. Large wasps were more likely to elicit a double cornicle secretion, indicating that aphids graded their response to the size of the attacker. Larger wasps were also more likely to be smeared with cornicle secretion, suggesting they were more vulnerable than small wasps. Although small wasps had smaller eggs than large wasps, there was no effect of maternal egg size on the size of progeny. However, daughters of small females emerged with larger egg loads than daughters of large mothers, and their eggs tended to be slightly smaller, although not significantly. Regression analysis revealed a positive correlation between maternal egg size and progeny developmental time for small and large wasps, and between maternal egg size and progeny egg load for small wasps. These results confirm maternal effects of body size in an aphid parasitoid, and reveal that vulnerability to host behavioral defenses is also body size dependent.     

Discrimination against previously searched, host-free patches by a parasitoid foraging for concealed hosts

1. The foraging behaviour of the parasitoid wasp Halticoptera laevigata Thomson (Hymenoptera: Pteromalidae) was studied. This wasp attacks the first‐instar larvae of the fruit fly Myoleja lucida Fallén (Diptera: Tephritidae) within honeysuckle Lonicera xylosteum L. (Caprifoliaceae) fruits. It was hypothesised that, to forage efficiently, wasps should be able to maintain a systematic or at least non‐random search pattern on the level of the microhabitat, i.e. branches with fruits, and/or be able to recognise previously searched fruits. 2. Comparisons of observed patch visitation patterns on branches bearing uninfested fruits with expected values for systematic and random visitation patterns indicated that H. laevigata wasps visit patches randomly and do not avoid visiting host‐free patches that they have visited previously. 3. While wasps did not alter their search effort on uninfested fruits searched previously by conspecifics, they searched for much shorter times and probed less frequently for hosts on uninfested fruits that they had searched previously. 4. The results indicate that H. laevigata wasps engage in partially systematic search and increase their search efficiency through the recognition of self‐searched fruits. The reasons why partial rather than fully systematic search should be adaptive in this parasitoid are discussed.     

Genetic and physiological data suggest demographic and adaptive responses in complex interactions between populations of figs (Ficus pumila) and their pollinating wasps (Wiebesia pumilae)

To study interactions between host figs and their pollinating wasps and the influence of climatic change on their genetic structures, we sequenced cytoplasmic and nuclear genes and genotyped nuclear microsatellite loci from two varieties of Ficus pumila, the widespread creeping fig and endemic jelly fig, and from their pollinating wasps, Wiebesia pumilae, found in Taiwan and on nearby offshore islands. Great divergence in the mitochondrial cytochrome c oxidase subunit I (mtCOI) with no genetic admixture in nuclear markers indicated that creeping‐ and jelly‐fig wasps are genetically distinct. Compared with creeping‐fig wasps, jelly‐fig wasps also showed better resistance under cold (20 °C) than warm (25 and 30 °C) conditions in a survival test, indicating their adaptation to a cold environment, which may have facilitated population expansion during the ice age as shown by a nuclear intron and 10 microsatellite loci. An excess of amino acid divergence and a pattern of too many rare mtCOI variants of jelly‐fig wasps as revealed by computer simulations and neutrality tests implied the effect of positive selection, which we hypothesize was associated with the cold‐adaptation process. Chloroplast DNA of the two fig plants was completely segregated, with signs of genetic admixture in nuclear markers. As creeping‐ and jelly‐fig wasps can pollinate creeping figs, occasional gene flow between the two figs is thus possible. Therefore, it is suggested that pollinating wasps may be playing an active role in driving introgression between different types of host fig.     

Bitrophic toxicity of Cry1Ac to Cycloneda sanguinea,a predator in Brazilian cotton

Insect predators are exposed to the Cry1Ac toxin in Bt cotton fields through several pathways. In this study, we investigated the effects of activated Cry1Ac added to a diet on Cycloneda sanguinea (L.) (Coleoptera: Coccinellidae), which is one of the main predators of non‐target pests in Brazilian cotton. Direct bitrophic exposure of C. sanguinea to Cry1Ac was done by feeding beetles with Aphis gossypii (Glover) (Hemiptera: Aphidae) sprayed with 500 μg per ml Cry1Ac solution. Larval and pupal survival, development time, aphid consumption, and adult longevity were recorded daily. Couples within the same experimental treatment were paired and numbers of eggs laid and hatched per female were recorded daily. Net replacement rate was calculated for each female. During development, a C. sanguinea larva consumed on average 1.8 μg of activated Cry1Ac. No significant differences due to Cry1Ac were observed for any of the response variables, except aphid consumption. Larvae receiving Cry1Ac consumed more aphids than larvae receiving distilled water alone. Additional statistical analyses were conducted to evaluate independence of responses, and for the independent responses, a simple meta‐analysis was conducted to test the null hypothesis that all responses were zero. Nearly all of the response variables were statistically independent. Two pairs of responses were not independent, but the associated multivariate tests were not significant. The meta‐analysis suggested that all effects were not different from random variation around zero and no cumulative effects could be detected. Our results indicated that bitrophic exposure to activated Cry1Ac is likely to have little or no adverse ecological effect on C. sanguinea.     

In vivo and in vitro developmental profiling of Asobara japonica,a larval endoparasitoid of drosophilid flies

The relationship between host and parasitoid has been examined in terms of either the effects of infestation on host growth and development, or the growth and development of parasitoid larvae in response to the physiological and nutritional status of their hosts. Although a wide range of host–parasitoid interactions has been studied, detailed developmental profiles of parasitoid larvae and environmental effects on them have remained unclear in many cases because the parasitoid larvae are relatively inaccessible inside their hosts. Here, we used Drosophila melanogaster Meigen (Diptera: Drosophilidae) and Asobara japonica Belokobylskij (Hymenoptera: Braconidae) as a model system to describe developmental profiles of hosts and endoparasitoid wasps, and investigated environmental factors that affect the developmental profiles in in vivo and in vitro culture experiments. As a result, we successfully identified six morphologically distinct developmental stages (I–VI) of pre‐adult A. japonica. The current approach based on qualitative and quantitative assessment of wasp morphology may be an effective approach for estimating the instar number of endoparasitoids lacking sclerotized structures in general. The finding that the development of A. japonica from stage III to stage IV is constrained by host developmental stage regardless of the environmental conditions in this study suggests that developmental mismatch may be an important factor in the evolution of host selection in endoparasitoid wasps.     

Overwintering strategy of endoparasitoids in Chilo suppressalis: a perspective from the cold hardiness of a host

Although parasitoids ultimately kill their host, koinobiont parasitoids must protect not only themselves but also their hosts against extreme environments. In this study, the parasitism rate of Chilo suppressalis Walker (Lepidoptera: Pyralidae) was investigated, and the average body weights, supercooling points, and concentrations of glycerol (acting as a cryoprotectant) in the hemolymph were compared between parasitized and non‐parasitized larvae. Five species of koinobiont endoparasitoids parasitized the overwintering C. suppressalis larvae and the total parasitism rate was 47.6% (n = 1 537). Average body weight of parasitized larvae was significantly lower than that of non‐parasitized larvae, and the parasitism rate of the lighter group (20–30 mg) was highest. The supercooling point of parasitized C. suppressalis larvae (?15.7 ± 0.3 °C) was significantly lower than that of the non‐parasitized larvae (?14.3 ± 0.2 °C). In addition, supercooling points were not correlated with body weights between parasitized and non‐parasitized larvae, indicating that cold hardiness of parasitized larvae was enhanced by endoparasitoids. Furthermore, the concentration of glycerol in the hemolymph was significantly higher in parasitized larvae (205.0 ± 7.1 μmol ml^?1) than in non‐parasitized larvae (169.8 ± 14.4 μmol ml^?1), which suggests that the mechanism that decreases the supercooling point of parasitized larvae was associated with glycerol. All these results indicated that the cold hardiness of parasitized C. suppressalis larvae was enhanced by their endoparasitoids, which benefitted overwintering endoparasitoids.     

Effects of starvation on the vitellogenesis,ovarian development and fecundity in the ectoparasitoid,Nasonia vitripennis (Hymenoptera: Pteromalidae)

A female‐specific protein, vitellogenin (Vg), and its corresponding egg vitellin (Vt) are identified in the ectoparasitic wasp Nasonia vitripennis. Both native Vt and Vg have a molecular mass of about 350 kDa, which is composed of two subunits of approximately 190 kDa and 165 kDa under reducing and denaturing conditions (sodium dodecyl sulfate—polyacrylamide gel electrophoresis). An indirect sandwich enzyme‐linked immunosorbent assay developed with both monoclonal and polyclonal antibodies against N. vitripennis Vt. Vg was first detected in the hemolymph on the 10th day after parasitism, and was first observed in oocytes on the 12th day. In adults deprived of food, the highest hemolymph Vg level occurred at the time of adult eclosion and the highest level of Vt in ovaries was found at 30 h after eclosion. In contrast, feeding adults with 20% sucrose resulted in the reduction of Vt uptake by ovaries and the extension of life span, but had little effect on Vg production. Deprived of hosts, starvation of female wasps had no significant effects on ovariole growth and oocyte maturation before the wasps died. However, starvation of female wasps supplied with hosts accelerated the wasps laying progeny into hosts, but resulted in a decrease of total progeny production by comparison with wasps fed with 20% sucrose.     

Effects of quantitative and qualitative differences in volatiles from host‐ and non‐host‐infested maize on the attraction of the larval parasitoid Cotesia kariyai

Cotesia kariyai Watanabe (Hymenoptera: Braconidae) is a specialist larval parasitoid of Mythimna separata Walker (Lepidoptera: Noctuidae). Cotesia kariyai wasps use herbivore‐induced plant volatiles (HIPVs) to locate hosts. However, complex natural habitats are full of volatiles released by both herbivorous host‐ and non‐host‐infested plants at various levels of intensity. Therefore, the presence of non‐hosts may affect parasitoid decisions while foraging. Here, the host‐finding efficiency of naive C. kariyai from HIPVs influenced by host‐ and non‐host‐infested maize [Zea mays L. (Poaceae)] plants was investigated with a four‐arm olfactometer. Ostrinia furnacalis Guenée (Lepidoptera: Crambidae) was selected as a non‐host species. One unit (1 U) of host‐ or non‐host‐infested plant was prepared by infesting a potted plant with five host or seven non‐host larvae. In two‐choice bioassays, host‐infested plants fed upon by different numbers of larvae, and various units of host‐ and non‐host‐infested plants (infestation units; 1 U, 2 U, and 3 U) were arranged to examine the effects of differences in volatile quantity and quality on the olfactory responses of C. kariyai with the assumption that volatile quantity and quality changes with differences in numbers of insects and plants. Cotesia kariyai was found to perceive quantitative differences in volatiles from host‐infested plants, preferring larger quantities of volatiles from larger numbers of larvae or plants. Also, the parasitoids discriminated between healthy plants, host‐infested plants, and non‐host‐infested plants by recognising volatiles released from those plants. Cotesia kariyai showed a reduced preference for host‐induced volatiles, when larger numbers of non‐host‐infested plants were present. Therefore, quantitative and qualitative differences in volatiles from host‐ and non‐host‐infested plants appear to affect the decision of C. kariyai during host‐habitat searching in multiple tritrophic systems.