Parasitoid vibrations as potential releasing stimulus of evasive behaviour in a leafminer

Abstract. The aim of this study was to characterize the vibratory signals produced by the parasitoid Sympiesis sericeicornis Nees (Hymenoptera: Eulophidae) while foraging on apple leaves infested by one of its hosts, the spotted tentiform leafminer Phyllonorycter malella (Ger.) (Lepidoptera: Gracillariidae).This leafminer changes its behaviour as a function of the parasitoid's behaviour to escape parasitization.We propose that the leafminer uses vibrations triggered by the parasitoid to detect the presence of its enemy.We measured vibrations produced by a foraging parasitoid on a mine with a laser vibrometer.By recording concurrently the behaviour of the parasitoid on video, vibrations could be assigned to particular behaviours.Subsequently, vibrations were characterized by their dominant frequencies and intensities.The behaviours Landing and Take-off both produced strong impact-like vibrations characterized by an initial irregular phase during which frequencies up to 25 kHz occurred followed by a slow decaying regular phase.Vibrations elicited by Moving, Standing and Probing showed no clear temporal pattern.During Probing , dominant frequencies of up to 5.6 kHz were observed frequently at intensities well above the background noise (>10 dB).During Moving and Standing , vibrations were more scarce and of lower frequencies and intensities.Due to their impact-like nature, vibrations produced by Landing and Take-off are probably not specific to the parasitoid.Vibrations produced by Moving and Standing are difficult to detect and not reliable because of their non-specificity.Therefore, only Probing provides a reliable and detectable source of information for the host.The vibrations elicited during Probing could account for the evasive behaviour that is observed in this and other leafminers.     

Matching host reactions to parasitoid wasp vibrations.

The sensory ecology of predator detection by prey has been little studied for any arthropod prey predator system, in contrast to the sensory ecology of prey finding by predators. The aim of this study was to quantify the foraging signals produced by the parasitoid Sympiesis sericeicornis (Hymenoptera: Eulophidae) and the sensory ecology of enemy detection and the avoidance behaviour of the leaf-mining host, Phyllonorycter spp. (Lepidoptera: Gracillariidae). We used synthetic vibrations approximating the signals generated by ovipositor insertions to stimulate the host in its mine. Tothe authors' knowledge, this is the first manipulative study to describe a match in the frequency range between a parasitoid foraging stimulus and a host behavioural response. We discuss our findings in relation to other predator-prey systems for which a coevolution between prey sensitivity and predator signal has been described.     

Host range of a newly introduced parasitoid, Binodoxys communis among common aphid species in Hawaii

Binodoxys communis (Gahan) (Hymenoptera:Braconidae), a parasitoid of aphids originally from China, was introduced into Hawaii and evaluated in the laboratory for its ability to detect, accept, oviposit and develop in Aphis gossypii reared on two host plants, plus five other common aphid species. The parasitoid was able to detect all six aphid species and to successfully sting five species, with highest preference for those in the genus Aphis. Aphis species were highly suitable for parasitoid development. Other species were only marginally suitable. Parasitoids spent less time searching on plants of less acceptable aphids. Aphid defensive behaviors did not affect oviposition success, but did lengthen the parasitoid’s handling time of several aphid species. Host acceptance was positively correlated with host suitability, yet one unsuitable host was readily accepted for oviposition.     

Temperature effects during development on a polyembryonic parasitoid and its host

Ambient temperature can influence development through effects on metabolic rate and by inducing physiological stress. In this study, we assessed temperature effects on a host–parasitoid interaction and on the body size and brood size of emerging wasps. By examining the development at two different temperatures of the koinobiont parasitoid, Copidosoma bakeri, and its host, Agrotis ipsilon, we asked: (1) Does the growth response to temperature by A. ipsilon depend on whether the moth caterpillar is parasitized? (2) Does the allocation pattern of body size and brood size in C. bakeri change with temperature? To answer these questions, we exposed A. ipsilon larvae parasitized by C. bakeri to high or low non-lethal temperatures when A. ipsilon was in early or late larval stages and measured their development time and body mass for all four treatment combinations. We also examined the brood size and body mass of emerging wasps. Whether parasitized or not, A. ipsilon larvae decreased development time, but generally did not decrease final body mass, at the higher temperature. When parasitized A. ipsilon was exposed to the higher temperature only late in their development, enlargement of the host by the parasitoid was reduced. C. bakeri brood size significantly increased when the higher temperature was applied early in host development. We did not detect a shift with temperature in the allocation pattern of the size–number trade-off for wasp offspring, suggesting that this trade-off relationship may be under selection strong enough to yield insensitivity to temperature.     

Virulence strategies in parasitoid Hymenoptera as an example of adaptive diversity

Parasitoids are mostly insects that develop at the expense of other arthropods, which will die as a result of the interaction. Their reproductive success thus totally depends on their ability to successfully infest their host whose reproductive success relies on its own ability to avoid or overcome parasitism. Such intense selective pressures have resulted in extremely diverse adaptations in parasitoid strategies that ensure parasitism success. For instance, wasp-specific viruses (polydnaviruses) are injected into the host by parasitoid females to modulate its physiology and immunity. This article synthesizes available physiological and molecular data on parasitoid virulence strategies and discusses the evolutionary processes at work. To cite this article: M. Poirié et al., C. R. Biologies 332 (2009).     

Habitats as Complex Odour Environments: How Does Plant Diversity Affect Herbivore and Parasitoid Orientation?

Plant diversity is known to affect success of host location by pest insects, but its effect on olfactory orientation of non-pest insect species has hardly been addressed. First, we tested in laboratory experiments the hypothesis that non-host plants, which increase odour complexity in habitats, affect the host location ability of herbivores and parasitoids. Furthermore, we recorded field data of plant diversity in addition to herbivore and parasitoid abundance at 77 grassland sites in three different regions in Germany in order to elucidate whether our laboratory results reflect the field situation. As a model system we used the herb Plantago lanceolata, the herbivorous weevil Mecinus pascuorum, and its larval parasitoid Mesopolobus incultus. The laboratory bioassays revealed that both the herbivorous weevil and its larval parasitoid can locate their host plant and host via olfactory cues even in the presence of non-host odour. In a newly established two-circle olfactometer, the weeviĺs capability to detect host plant odour was not affected by odours from non-host plants. However, addition of non-host plant odours to host plant odour enhanced the weeviĺs foraging activity. The parasitoid was attracted by a combination of host plant and host volatiles in both the absence and presence of non-host plant volatiles in a Y-tube olfactometer. In dual choice tests the parasitoid preferred the blend of host plant and host volatiles over its combination with non-host plant volatiles. In the field, no indication was found that high plant diversity disturbs host (plant) location by the weevil and its parasitoid. In contrast, plant diversity was positively correlated with weevil abundance, whereas parasitoid abundance was independent of plant diversity. Therefore, we conclude that weevils and parasitoids showed the sensory capacity to successfully cope with complex vegetation odours when searching for hosts.     

Resource dependence in a new ecosystem: A host plant and its colonizing community

The introduced black locust (Robinia pseudoacacia) has become an invasive plant species in Europe. The introduction of alien plants such as the black locust may modify ecosystem composition and functioning. In response to the presence of a potential host plant, herbivores can adapt and shift to the consumption of the new host plant. In Eastern-Central Europe, the seed predator Bruchophagus robiniae (Hymenoptera: Eurytomidae) is an important consumer of black locust seeds which presumably shifted from its formerly host species to black locust. We tested the influence of host plant abundance on a seed predator – parasitoid community. We found that the seed predator B. robiniae was present in higher numbers in woodlots than in small patches of black locust. The density of the specialist parasitoid Mesopolobus sp. was lower in woodlots than in small patches, while the generalist parasitoid Eupelmus urozonos was evenly distributed between woodlots and small patches of black locust. We found that parasitoid species are influenced by the patch size of host plants, thus characteristics of introduced host plants can also manifest in higher trophic levels.     

Insights to host discrimination and host acceptance behaviour in a parasitoid (Diptera: Asilidae): Implications for fitness

The robber fly Mallophora ruficauda is one of the principal pests of apiculture in the Pampas region of Argentina. As adults they prey on honey bees and other insects, while as larvae they are solitary ectoparasitoids of third instar scarab beetle larvae. Females of M. ruficauda lay eggs away from the host in tall grasses. After being dispersed by the wind, larvae drop to the ground, where they dig in search of their hosts. It is known that second instar larvae of M. ruficauda exhibit active host searching behaviour towards its preferred host, third instar larva of Cyclocephala signaticollis. Although the means by which host location occurs has been studied and since superparasitism is a frequent scenario in the field, no information about host discrimination and host acceptance is available. We carried out studies in the field and behavioural experiments in the laboratory to determine if M. ruficauda is capable of quality host discrimination. We also studied if this parasitoid is capable of conspecific detection in order to avoid superparasitism. Finally, we analyzed the conditions under which superparasitism occurs in the field. We report here that the second instar larva of M. ruficauda is able to discriminate the parasitism status of the host by means of chemical cues, but is not capable of detecting conspecifics prior to attacking a host. We also found that the host cannot detect the presence of the parasitoid by means of chemical cues, so that no counter-defense against parasitism occurs. Furthermore, we determined that superparasitism occurs on the heavier hosts, i.e. those with more abundant resources which could harbor several parasitoid individuals. Finally, we discuss the possible implications of larval host location and host discrimination decisions on the fitness of this parasitoid.     

Sex ratios in field populations of two parasitoids (Hymenoptera: Chalcidoidea) of Coccus hesperidum L. (Homoptera: Coccidae)

We tested several assumptions and predictions of host-quality-dependent sex allocation theory (Charnov et al. 1981) with data obtained for the parasitoid Metaphycus stanleyi Compere on its host, brown soft scale (Coccus hesperidum L.), in a California citrus grove and in the laboratory. Scales ceased growing after parasitization by M.?stanleyi. Thus, M. stanleyi may gauge host quality (=size) at oviposition. Host size positively influenced adult parasitoid size, and parasitoid size in turn influenced adult longevity of M. stanleyi. However, parasitoid fitness gains with host size and adult size were similar in males versus females. Sex allocation to individual hosts by M. stanleyi depended on host size; females consistently emerged from larger hosts than males. Host size was important in a relative sense; the mean host sizes of females versus males, and of solitary versus gregarious parasitoids varied with the available host size distribution. The offspring sex ratio of M. stanleyi reflected the available host size distribution; the sex ratio of emerging parasitoids varied with the available host size distribution. We did not detect a “critical host size” below which males emerged, and above which females emerged; rather, only females emerged from hosts in the upper size range, and a variable ratio of males and females emerged from hosts in the lower size range. We conclude that the sex ratio of field populations of M.?stanleyi is driven largely by the available size distribution of C. hesperidum. In addition, we tested predictions resulting from theoretical analyses of sex allocation in autoparasitoids with data obtained on Coccophagus semicircularis (Förster) parasitizing brown soft scale in the field. The sex ratio of C. semicircularis was consistently and strongly female biased (ca. 90% females). Based on available theoretical analyses, we suggest that this sex ratio pattern may have resulted from a very low encounter rate of secondary hosts coupled with a strong time limitation in C. semicircularis females. This explanation was the most plausible given constraints stemming from the detection of secondary hosts, their variable location within primary hosts, and their handling times. Finally, the size of hosts which yielded single versus multiple parasitoids, and the sizes of these parasitoids, were compared. These comparisons suggested that: (1) M. stanleyi females gauge host sizes precisely, and in terms of female offspring; thus a fitness penalty is not incurred by females which share a host, while males benefit from sharing a host, and; (2) instances where multiple C. semicircularis emerged from a single host were probably the result of parasitism by different females, or during different encounters by a single female.     

Larvae of the tachinid fly,Drino inconspicuoides (Diptera: Tachinidae), suppress melanization in host lepidopteran insects

The ovoviviparous parasitoid, Drino inconspicuoides (Diptera: Tachinidae) parasitizes a wide range of lepidopteran insects; larval period progresses in the host hemocoel. Here, we examined how D. inconspicuoides responds to melanization, which involves the activation of prophenoloxidases and is the first immune reaction induced by the host against invading organisms. We found that the larvae of D. inconspicuoides suppressed the activation of prophenoloxidases in its natural hosts, Mythimna separata (Lepidoptera: Noctuidae) and Bombyx mori (Lepidoptera: Bombycidae). The suppression of melanization starts immediately after invasion and is maintained for at least 24 h. We did not detect a drastic degradation of prophenoloxidases, suggesting that the presence of other molecules targeted by D. inconspicuoides suppresses melanization. D. inconspicuoides does not inhibit a cellular immune reaction, encapsulation, and thus, it is likely that the tachinids survive secondary infections of the host by partially retaining the host immune function.     

The influence of the rearing host on the response of the parasitoid Venturia canescens (Gravenhorst) (Hymenoptera: Ichneumonidae) to odours from Ephestia kuehniella and Plodia interpunctella in a Y-tube olfactometer

Venturia canescens (Gravenhorst) is an ichneumonid generalist parasitoid that successfully attacks the larvae of different lepidopteran pests that infest stored products. These pest species include Plodia interpunctella and Ephestia kuehniella. In this study, we aimed to evaluate the influence of the rearing host on the parasitoid’s ability to detect and respond to a new host different from the rearing species. For this reason, the trials tested the preference of parasitoids reared on P. interpunctella or E. kuehniella for products that were or were not infested with larvae of these hosts. The trials were conducted in a Y-tube olfactometer. Regardless of the rearing host species, the parasitoids showed no preference for uninfested products. The parasitoids were attracted to products infested with larvae of their rearing host in preference to uninfested products. They also showed preferential attraction to products infested with the new host over uninfested products. E. kuehniella was the preferred host, irrespectively of the parasitoid host rearing species. The results are discussed to develop a better understanding of the ecology of V. canescens for its application in biological control.     

Manipulation of parasitoid state influences host exploitation by Diadromus pulchellus Wesmael (Hymenoptera: Ichneumonidae)

In biological control programs, it should be possible to manipulate agents during host specificity trials so that they exhibit a lower host acceptance threshold. This would help to ensure that any non-target species that could be attacked in the field will also be attacked in the laboratory and flagged as potential hosts. The question is how to encourage agents to express the widest possible host range during trials. This study examined the effects of variation in parasitoid state (mating status, nutritional status, age, host experience) and the test environment (competition, arena size, presence of host-damaged leek leaves) on the degree of host exploitation by Diadromus pulchellus Wesmael (Hymenoptera: Ichneumonidae). As a first step, this study looked for changes in oviposition by the parasitoid on its normal host, the leek moth, Acrolepiopsis assectella (Zeller) (Lepidoptera: Acrolepiidae). Each of the seven factors was tested independently to assess its effect on the number of parasitoid offspring produced and host mortality. Previous host experience led to a marginal increase, while competition led to a per-capita decrease, in the degree of host exploitation. In contrast, mating status, arena size and the presence of host-damaged leek leaves had no measurable effects. Finally, parasitoid age and nutritional status had strong effects in the opposite direction to predictions, with younger and sugar-fed individuals exhibiting higher host exploitation than older and sugar-starved parasitoids, respectively. This study suggests that a sound understanding of species-specific reproductive biology and parasitoid–host interactions is likely required before oviposition-enhancing parasitoid states can confidently be identified.     

Interactions between ant species increase arrival rates of an ant parasitoid

Behaviours or traits associated with aggression and communication may increase an animal's conspicuousness to predators or parasitoids. Most examples of this come from instances of aggression or communication within a species. We tested whether interspecific encounters between ants enhance the host location success of a parasitoid (Diptera: Phoridae) that attacks ants in the genus Linepithema. At food resources recruited to by Linepithema, parasitoid discovery rates were lower when Linepithema was alone than when other ant species were present. In experimentally controlled encounters, parasitoid discovery rates were elevated when Linepithema confronted an ant species that elicited use of chemicals, but not when it confronted an ant species that primarily elicited physical aggression. These results indicate that phorid parasitoids of Linepithema use the ant's chemicals as host location cues. Because Linepithema is known to abandon food resources in the presence of its phorid parasitoids, its use of chemicals during interspecific encounters may diminish its competitive success when phorids are nearby. Copyright 2003 Published by Elsevier Science Ltd on behalf of The Association for the Study of Animal Behaviour.      

Does variation in host plant association and symbiont infection of pea aphid populations induce genetic and behaviour differentiation of its main parasitoid, Aphidius ervi?

The host-associated differentiation (HAD) hypothesis states that higher trophic levels in parasitic associations should exhibit similar divergence in case of host sympatric speciation. We tested HAD on populations of Aphidius ervi the main parasitoid of the pea aphid Acyrthosiphon pisum, emerging from host populations specialized on either alfalfa or red clover. Host and parasitoid populations were assessed for genetic variation and structure, while considering geography, host plant and host aphid protective symbionts Regiella insecticola and Hamiltonella defensa as potential covariables. Cluster and hierarchical analyses were used to assess the contribution of these variables to population structure, based on genotyping pea aphids and associated A. ervi with microsatellites, and host aphid facultative symbionts with 16S rDNA markers. Pea aphid genotypes were clearly distributed in two groups closely corresponding with their plant origins, confirming strong plant associated differentiation of this aphid in North America. Overall parasitism by A. ervi averaged 21.5 % across samples, and many parasitized aphids producing a wasp hosted defensive bacteria, indicating partial or ineffective protective efficacy of these symbionts in the field. The A. ervi population genetic data failed to support differentiation according to the host plant association of their pea aphid host. Potential for parasitoid specialization was also explored in experiments where wasps from alfalfa and clover aphids were reciprocally transplanted on alternate hosts, the hypothesis being that wasp behaviour and parasitic stages should be most adapted to their host of origin. Results revealed higher probability of oviposition on the alfalfa aphids, but higher adult emergence success on red clover aphids, with no interaction as expected under HAD. We conclude that our study provides no support for the HAD in this system. We discuss factors that might impair A. ervi specialization on its divergent aphid hosts on alfalfa and clover.     

Electroantennogram (EAG) responses of Microplitis croceipes and Cotesia marginiventris and their lepidopteran hosts to a wide array of odor stimuli: Correlation between EAG response and degree of host specificity?

In order to test whether the electroantennogram (EAG) response spectrum of an insect correlates to its degree of host specificity, we recorded EAG responses of two parasitoid species with different degrees of host specificity, Microplitis croceipes (specialist) and Cotesia marginiventris (generalist), to a wide array of odor stimuli including compounds representing green leaf volatiles (GLVs), herbivore-induced plant volatiles (HIPV), ecologically irrelevant (not used by the parasitoid species and their hosts for host location) plant volatiles, and host-specific odor stimuli (host sex pheromones, and extracts of host caterpillar body and frass). We also tested the EAG responses of female moths of the caterpillar hosts of the parasitoids, Heliothis virescens and Spodoptera exigua, to some of the odor stimuli. We hypothesized that the specialist parasitoid will have a narrower EAG response spectrum than the generalist, and that the two lepidopteran species, which are similar in their host plant use, will show similar EAG response spectra to plant volatiles. As predicted, the specialist parasitoid showed greater EAG responses than the generalist to host-specific odor and one HIPV (cis-3-hexenyl butyrate), whereas the generalist showed relatively greater EAG responses to the GLVs and unrelated plant volatiles. We detected no differences in the EAG responses of H. virescens and S. exigua to any of the tested odor.     

Substrate vibrations elicit defensive behaviour in leafminer pupae

Late instar larvae and pupae of the spotted tentiform leafminer Phyllonorycter malella (Ger.) (Lepidoptera: Gracillariidae) react with defensive behaviour when attacked by one of their parasitoids, the eulophid wasp Sympiesis sericeicornis Nees (Hymenoptera: Eulophidae). Vibrations produced during the insertion of the ovipositor into the mine are known to be important cues by which larvae detect the presence of their enemies. The aim of this study was to investigate which frequency components elicit defensive reactions in leafminer pupae using synthetic vibrations. Sine vibrations and bandlimited noise stimuli were offered to both free pupae and pupae concealed in their leafmines. Using Laser vibrometry we measured the vibrations experienced by pupae inside their mines and assessed the influence of the mine. Pupae were shown to react to substrate vibrations, and do so over a broad range of frequencies. Behavioural reactions to noise stimuli were stronger than to pure sine stimuli. Mine tissue attenuated vibration amplitudes of the input signal from 5.1 to 22.6dB. However, as response thresholds of concealed pupae were only twice as high as thresholds of free pupae (which is adequate to 3dB) pupae inside their mine were more sensitive than expected. This discrepancy is discussed both in terms of the conditions of pupae and in terms of mine structure. The results indicate that broadbandedness of vibrations produced by hunting parasitoids during ovipositor insertion into the mine may be a major criterion used by leafminers to perceive parasitoid presence and to escape ovipositor stings.     

Who is the puppet master? Replication of a parasitic wasp-associated virus correlates with host behaviour manipulation

Many parasites modify their host behaviour to improve their own transmission and survival, but the proximate mechanisms remain poorly understood. An original model consists of the parasitoid Dinocampus coccinellae and its coccinellid host, Coleomegilla maculata; during the behaviour manipulation, the parasitoid is not in contact with its host anymore. We report herein the discovery and characterization of a new RNA virus of the parasitoid (D. coccinellae paralysis virus, DcPV). Using a combination of RT-qPCR and transmission electron microscopy, we demonstrate that DcPV is stored in the oviduct of parasitoid females, replicates in parasitoid larvae and is transmitted to the host during larval development. Next, DcPV replication in the host''s nervous tissue induces a severe neuropathy and antiviral immune response that correlate with the paralytic symptoms characterizing the behaviour manipulation. Remarkably, virus clearance correlates with recovery of normal coccinellid behaviour. These results provide evidence that changes in ladybeetle behaviour most likely result from DcPV replication in the cerebral ganglia rather than by manipulation by the parasitoid. This offers stimulating prospects for research on parasitic manipulation by suggesting for the first time that behaviour manipulation could be symbiont-mediated.     

Influence of “protective” symbionts throughout the different steps of an aphid–parasitoid interaction

Microbial associates are widespread in insects, some conferring a protection to their hosts against natural enemies like parasitoids. These protective symbionts may affect the infection success of the parasitoid by modifying behavioral defenses of their hosts, the development success of the parasitoid by conferring a resistance against it or by altering life-history traits of the emerging parasitoids. Here, we assessed the effects of different protective bacterial symbionts on the entire sequence of the host-parasitoid interaction (i.e., from parasitoid attack to offspring emergence) between the pea aphid, Acyrthosiphon pisum, and its main parasitoid, Aphidius ervi and their impacts on the life-history traits of the emerging parasitoids. To test whether symbiont-mediated phenotypes were general or specific to particular aphid–symbiont associations, we considered several aphid lineages, each harboring a different strain of either Hamiltonella defensa or Regiella insecticola, two protective symbionts commonly found in aphids. We found that symbiont species and strains had a weak effect on the ability of aphids to defend themselves against the parasitic wasps during the attack and a strong effect on aphid resistance against parasitoid development. While parasitism resistance was mainly determined by symbionts, their effects on host defensive behaviors varied largely from one aphid–symbiont association to another. Also, the symbiotic status of the aphid individuals had no impact on the attack rate of the parasitic wasps, the parasitoid emergence rate from parasitized aphids nor the life-history traits of the emerging parasitoids. Overall, no correlations between symbiont effects on the different stages of the host–parasitoid interaction was observed, suggesting no trade-offs or positive associations between symbiont-mediated phenotypes. Our study highlights the need to consider various sequences of the host-parasitoid interaction to better assess the outcomes of protective symbioses and understand the ecological and evolutionary dynamics of insect–symbiont associations.     

Effects of rearing conditions on the parasitism of Tetrastichus brontispae on its pupal host Octodonta nipae

Octodonta nipae (Maulik) (Coleoptera: Chrysomelidae) is currently a serious invasive pest of palm plants in southern China. Although previous studies reported that Tetrastichus brontispae Ferriere (Hymenoptera: Eulophidae) could successfully parasitize the pupae of O. nipae, little is known of the potential for T. brontispae to control this pest. To understand the interaction between T. brontispae and O. nipae, the effects of parasitoid feeding regimes, host age and density, parasitoid age and density, and cold storage on the parasitism of T. brontispae were investigated. Our results indicate that a supply of either glucose or sucrose increased the longevity and fecundity of T. brontispae. The parasitoid preferred one-day-old host pupae and its fecundity increased with increasing host or parasitoid female density. Extended cold storage negatively affected the parasitism of T. brontispae. These results demonstrate how to efficiently raise T. brontispae and will contribute to the effective management of O. nipae.