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.     

Complex feeding tracks of the sessile herbivorous insect Ophiomyia maura as a function of the defense against insect parasitoids

Because insect herbivores generally suffer from high mortality due to their natural enemies, reducing the risk of being located by natural enemies is of critical importance for them, forcing them to develop a variety of defensive measures. Larvae of leaf-mining insects lead a sedentary life inside a leaf and make conspicuous feeding tracks called mines, exposing themselves to the potential risk of parasitism. We investigated the defense strategy of the linear leafminer Ophiomyia maura Meigen (Diptera: Agromyzidae), by focusing on its mining patterns. We examined whether the leafminer could reduce the risk of being parasitized (1) by making cross structures in the inner area of a leaf to deter parasitoids from tracking the mines due to complex pathways, and (2) by mining along the edge of a leaf to hinder visually searching parasitoids from finding mined leaves due to effective background matching of the mined leaves among intact leaves. We quantified fractal dimension as mine complexity and area of mine in the inner area of the leaf as interior mine density for each sample mine, and analyzed whether these mine traits affected the susceptibility of O. maura to parasitism. Our results have shown that an increase in mine complexity with the development of occupying larvae decreases the probability of being parasitized, while interior mine density has no influence on parasitism. These results suggest that the larval development increases the host defense ability through increasing mine complexity. Thus the feeding pattern of these sessile insects has a defensive function by reducing the risk of parasitism.     

Discrimination by Coptera haywardi (Hymenoptera: Diapriidae) of hosts previously attacked by conspecifics or by the larval parasitoid Diachasmimorpha longicaudata (Hymenoptera: Braconidae)

Coptera haywardi (Oglobin) is an endoparasitoid of fruit fly pupae that could find itself in competition with other parasitoids, both con- and heterospecific, already resident inside hosts. In choice bioassays, ovipositing C. haywardi females strongly discriminated against conspecifically parasitised Anastrepha ludens (Loew) pupal hosts. They also avoided pupae previously attacked by Diachasmimorpha longicaudata (Ashmead), a larval–prepupal koinobiont endoparasitoid, and the degree of larval-parasitoid superparasitism had no effect on this avoidance. There was no difference in the number of ovipositor insertions when hosts previously parasitised by a conspecific and D. longicaudata were exposed simultaneously. As females aged the degree of host discrimination declined. An ability to discriminate against pupae previously attacked as larvae suggests low levels of both conspecific and heterospecific competition in the field.     

Importance of contact chemical cues in host recognition and acceptance by the braconid larval endoparasitoids Cotesia sesamiae and Cotesia flavipes

The ability of the congeneric braconid parasitoids Cotesia sesamiae (Cameron) and Cotesia flavipes Cameron to discriminate between stemborer larval cues upon contact was studied using their natural hosts, namely the noctuid Busseola fusca (Fuller) and the crambid Chilo partellus (Swinhoe), respectively, and the pyralid non-host Eldana saccharina (Walker). When the natural host larvae were washed in distilled water, parasitoid behavior was similar to that displayed when in contact with E. saccharina, characterized by the absence of ovipositor insertion. When washed host or non-host larvae were bathed with water extracts of their natural host, the parasitoids showed a significant increase in ovipositor insertions. However, the water extracts of host-larvae deposited on cotton wool balls did not induce ovipositor insertion in either C. sesamiae or C. flavipes. Nevertheless, the extracts enabled the parasitoids to discriminate between natural and non-hosts as indicated by the intensive antennating of the former. For both parasitoids, frass was found to be important in short-range host recognition as indicated by differences in the time spent on antennating between frass sources. In addition, the regurgitants of B. fusca and C. partellus induced ovipositor insertion in C. flavipes only. These results indicated that C. sesamiae and C. flavipes used different chemical cues for acceptation and oviposition in a stemborer larva, and that B. fusca and C. partellus shared the same chemical cues to induce oviposition in C. flavipes.     

Host location by a parasitoid using leafminer vibrations: characterizing the vibrational signals produced by the leafmining host

Abstract. The aim of this study was to characterize the vibrations produced by the apple tentiform leafminer Phyllonorycter malella (Ger.) (Lepidoptera, Gracillariidae). Host location using vibrations by one of its parasitoids Sympiesis sericeicornis Nees (Hymenoptera, Eulophidae) was postulated by Casas (1989) on the basis of detailed quantitative behavioural observations and has also been suggested by other authors on similar systems. Both host and parasitoid send and may receive vibratory signals; consequently we first attempted to characterize and classify the signals, one of the first steps required in the design of an adequate vibrational biotest. In this respect, our approach differs fundamentally from the familiar setting of host location via semiochemicals and is best framed within the context of vibratory communication. Vibrational signals produced by a moving larva and pupa were measured on several spots on the leaf using a laser vibrometer. The emitted signals were characterized by their temporal patterns of change in amplitude and frequency spectra. The vibrational patterns released by a moving larva were different from those released by a wriggling larva and a wriggling pupa in the time as well as in the frequency domains. Wriggling larvae and wriggling pupae triggered vibrations that were similar in frequency, but differed in their temporal pattern. Frequencies up to 15 kHz could be identified. The amplitudes and frequencies of the signals both decreased significantly from the tip to the base of the leaf. A wriggling pupa and a wriggling larva produced stronger signals than a foraging larva. All calculated parameters (displacement, velocity, acceleration, and duration of the signal components) of the vibrational signals were found to be in a range comparable with others used for well-known arthropod communication systems. The vibrations produced by the host displayed distinct characteristics: they could usually be distinguished easily from background noise; could be perceived anywhere on the leaf; and were specific for a certain host stage and activity. Our findings support the hypothesis that vibration signals represent a reliable source of information to foraging parasitoids and, therefore, explain certain behavioural patterns observed in a population of S. sericeicornis females foraging in the field.     

Superparasitism in mass reared Diachasmimorpha longicaudata (Ashmead) (Hymenoptera: Braconidae), a parasitoid of fruit flies (Diptera: Tephritidae)

Superparasitism frequency and its effects on the quality of mass-reared Diachasmimorpha longicaudata (Ashmead) parasitoids were investigated under laboratory conditions. Percentage of adult emergence, sex ratio, survival, fecundity and flight ability of adult parasitoids that emerged from Anastrepha ludens (Loew) pupae with different levels of superparasitism were determined. A high prevalence of superparasitism was observed. The number of scars per pupa, produced by insertion of the parasitoid ovipositor, ranged from 1 to 30, with an average (±SD) of 8.3 ± 6.2. Adult parasitoid emergence decreased as the level of superparasitism increased. However, the fraction of females rose with increasing superparasitism and the flight ability was lower in adults emerging from pupae with only one scar, compared with adults emerging from superparasitized hosts. Female longevity and fecundity were not affected by superparasitism. Our results support the hypothesis that superparasitism in D. longicaudata might be adaptive, since adults emerging from hosts with moderate levels of superparasitism showed the highest percentage of emergence and there were no significant differences in the other quality control parameters tested. Our findings are relevant to the mass rearing process, where the ratio of hosts to parasitoids can be optimized as well as the distribution of eggs deposited in host larvae. This contributes to efficient mass rearing methods for augmentative biological control programs.     

A compound produced by fruigivorous Tephritidae (Diptera) larvae promotes oviposition behavior by the biological control agent Diachasmimorpha longicaudata (Hymenoptera: Braconidae)

Tephritid fruit fly parasitoids use fruit-derived chemical cues and the vibrations that result from larval movements to locate hosts sequestered inside fruit. However, compounds produced by the larvae themselves have not been previously described nor their significance to parasitoid foraging determined. We collected the volatiles from four species of tropical and subtropical Tephritidae: Anastrepha suspensa (Loew), Bactrocera dorsalis Hendel, Bactrocera cucurbitae Coquillett, and Ceratitis capitata (Wiedemann), representing two subfamilies (Dacinae and Trypetinae). Para-ethylacetophenone, an analog of a known tephritid parasitoid attractant, was a major constituent of all four, and was not associated with larvae of another acalypterate fly, Drosophila melanogaster Meigen, or with the calypterate Musca domestica L. It also was present in volatiles from whole, A. suspensa infested fruits of Eugenia uniflora (L.). Para-ethylacetophenone was not necessarily produced as a direct consequence of fruit consumption because it also was detected from larvae that developed in two artificial diets and in spent diets subsequent to larval development. Sensillae on both the antennae and ovipositor of the opiine braconid fruit fly parasitoid, Diachasmimorpha longicaudata (Ashmead) responded to the para-ethylacetophenone in larval volatiles and as a synthetic. Although a potential cue to foraging parasitoids, para-ethylacetophenone showed no long range (>1m) attractiveness to the adult female parasitoid, but did stimulate ovipositor-insertion and oviposition into both a natural (fruit) and an artificial (parafilm) substrate. Thus it may prove useful in colonizing and mass-rearing opine fruit fly parasitoids.     

Efficiency of vibrational sounding in parasitoid host location depends on substrate density

Parasitoids of concealed hosts have to drill through a substrate with their ovipositor for successful parasitization. Hymenopteran species in this drill-and-sting guild locate immobile pupal hosts by vibrational sounding, i.e., echolocation on solid substrate. Although this host location strategy is assumed to be common among the Orussidae and Ichneumonidae there is no information yet whether it is adapted to characteristics of the host microhabitat. This study examined the effect of substrate density on responsiveness and host location efficiency in two pupal parasitoids, Pimpla turionellae and Xanthopimpla stemmator (Hymenoptera: Ichneumonidae), with different host-niche specialization and corresponding ovipositor morphology. Location and frequency of ovipositor insertions were scored on cylindrical plant stem models of various densities. Substrate density had a significant negative effect on responsiveness, number of ovipositor insertions, and host location precision in both species. The more niche-specific species X. stemmator showed a higher host location precision and insertion activity. We could show that vibrational sounding is obviously adapted to the host microhabitat of the parasitoid species using this host location strategy. We suggest the attenuation of pulses during vibrational sounding as the energetically costly limiting factor for this adaptation.     

Host selection by Ibalia leucospoides based on temporal variations of volatiles from the hosts’ fungal symbiont

Insect parasitoids locate hosts via reliable and predictable cues such as volatile emissions from hosts and/or host plants. For insects that depend on mutualistic organisms, such as many wood‐boring insects, symbiont‐derived semiochemicals may represent a source of such cues to be exploited by natural enemies. Ultimately, exploitation of these signals may increase fitness by optimizing foraging efficiency. Female parasitoids of Ibalia leucospoides use volatiles from the fungal symbiont Amylostereum areolatum of their host Sirex noctlio to find concealed host eggs and young larvae within the xylem. We hypothesize that the temporal pattern of fungal emissions may indicate not only the presence of host larvae but also be used as a cue that indicates host suitability and age. Such information would allow female parasitoids to discern more efficiently between hosts within ovipositor reach from those already buried too deep into the xylem and out of reach. In this context, we assessed the behaviour of I. leucospoides females to volatiles of A. areolatum in a ‘Y’‐tube olfactometer at regular intervals over 30 days. We concurrently examined the fungal volatiles by headspace sampling through solid‐phase microextraction (SPME) followed by gas chromatography mass spectrometry (GC‐MS). We observed that females were attracted to volatiles produced by two‐week‐old fungal cultures, a period that matches when older larvae are still within ovipositor reach. Four chemical compounds were detected: ethanol, acetone, acetaldehyde and the sesquiterpene 2,2,8‐trimethyltricyclo[6.2.2.01,6]dodec‐5‐ene, with each compounds’ relative abundance changing over time. Results are discussed in the context of parasitoids fitness. Future studies involving electrophysiology, different collection techniques and further behavioural assays will help in identifying the compounds that convey temporal information to female parasitoids and have the potential for being used in integrated pest management programmes.     

Host-handling behavior of the parasitoid,Diglyphus minoeus [Hym.: Eulophidae], parasitizing the honeysuckle leaf-miner,Phytomyza lonicerae [Dipt.: Agromyzidae]

The host-handling behavior of individual female waspsDiglyphys minoeus which attack the larvae of the honeysuckle leaf-miner,Phytomyza lonicerae, include 4 types of behavior: probing, ovipositor insertion, host-feeding, and resting. Ovipositor insertion may either be injection of venom, probing of the host by the sensilla on the ovipositor's tip or egg laying. Three types of attacks were distinguished: oviposition attack for unparasitized larvae, host-feeding attack for unparasitized host larvae and host rejection for previously parasitized host larvae. Oviposition attack was characterized by frequent alternation between probing and ovipositor insertion, long duration of ovipositor insertion and resting, and the long duration of host-handling. Resting behavior is thought to protect the progeny against superparasitism, host-feeding by other wasps or hosts' recovery from paralysis. Host-feeding attack was characterized by frequent alternation between host-feeding and ovipositor insertion and long duration of host-feeding. Host rejection was composed mainly of probing and ovipositor insertion and short handling time.      

Juvenile and sublethal effects of selected pesticides on the leafminer parasitoids Hemiptarsenus varicornis and Diglyphus isaea (Hymenoptera: Eulophidae) from Australia

The pest leafminers Liriomyza huidobrensis (Blanchard), Liriomyza sativae (Blanchard), and Liriomyza trifolii (Burgess) (Diptera: Agromyzidae) have spread into South East Asia and Oceania, and they are likely to reach Australia in the near future. Two translaminar pesticides, cyromazine and abamectin, currently provide effective chemical control of these pests, but because parasitoids can play an important role in controlling and preventing leafminer outbreaks, understanding the impact of pesticides on leafminer parasitoids is vital. Here, we tested larval and pupal mortality and sublethal effects of abamectin, cyromazine, and the widely used fungicide mancozeb on two common Australian leafminer parasitoids, Hemiptarsenus varicornis (Girault) and Diglyphus isaea (Walker). Abamectin caused significant mortality to larvae and pupae of both parasitoid species but cyromazine and mancozeb did not. Progeny production and longevity of H. varicornis were not affected by adult exposure to cyromazine and mancozeb, nor did direct pupal exposure decrease number of progeny produced by either parasitoid. Mortality of H. varicornis females emerging from leaves treated with abamectin was high for up to 72 h after eclosion but those surviving beyond 72 h did not differ from control females in the number of progeny produced. Mancozeb did not influence leaf residence time or parasitism by H. varicornis females. Cyromazine and the fungicide mancozeb were concluded to be compatible with the parasitoids tested and suitable for integrated pest management of leafminers should outbreaks of pest species occur in Australia. Abamectin should be used with caution because it caused significant mortality in both parasitoids tested here.     

Bionomics and morphological and molecular characterization of Elasmus schmitti and Baryscapus elasmi (Hymenoptera: Chalcidoidea, Eulophidae), parasitoids associated with a paper wasp, Polistes dominulus (Vespoidea, Vespidae)

Elasmus schmitti and Baryscapus elasmi have been recorded in southern Ukraine as gregarious parasitoids in the nests of the paper wasps Polistes dominulus and Polistes nimphus. Polistes dominulus nests infested with E. schmitti were less productive than uninfested nests in only one year (2004) of the three years of the present study, when an increase in the host population size occurred. Females of E. schmitti are synovigenic, and they lay their eggs on the skins of P. dominulus last instar larvae, without paralyzing the host. Rather, the parasitoid larvae feed on young host pupae. The pupae of E. schmitti are isolated from the host remnants by a thin fecal partition as in Elasmus polistis and Elasmus japonicus, other paper wasp parasitoids. Baryscapus elasmi is a pupal endoparasitoid of E. schmitti. The females of B. elasmi emerge without mature eggs in their ovaries and mate with males. They penetrate the paper wasp’s cells with their ovipositor and feed on the extracted hemolymph exudate. Pupation of B. elasmi occurs inside or outside the pupa of the host, E. schmitti. If inside, then the cranial end of the pupa and the adult emergence hole of B. elasmi are situated in the caudal ends of the pupae of their hosts. Comparative notes and illustrations on the morphology of adults are provided, and DNA sequences of three genes (nuclear 28S D2 rDNA, mitochondrial cytochrome oxidase subunit I, and mitochondrial cytochrome b) were obtained for both parasitoid species. The similarity of the 28S D2 sequences of E. schmitti and E. polistis relative to other available Elasmus sequences suggests a single origin of parasitism on paper wasps in this genus.     

Behavioral mechanisms of parasitic wasps for searching concealed insect hosts

Wood borers are important forest insect pests and difficult to be controlled owing to their concealed behavior. However, parasitic wasps can effectively ascertain and parasitize wood borers as well as other concealed pests by using special searching, finding and attacking mechanisms, which have been developed during the course of long-term coevolution with their hosts. The present paper summarizes the behavioral mechanisms of parasitic wasps involved in searching and locating their concealed hosts. Parasitic wasps can accurately find the location of their hidden hosts and then parasitize them, usually by using olfactory semiochemicals from hosts (lavare and adults), host frass and symbiotic microorganisms in host galleries; visual signals from color contrast of plant surface; contact stimuli from characters of host physical defense; substrate vibrations produced by host feeding and movement; infrared radiation from host activities and metabolizability. Some parasitic wasps may integrate the information of several stimuli from different sources to enhance the reliability and accuracy of host locations. In addition, the potentials for utilizing the host location signals of parasitoids in biological control are discussed.     

Influence of experience on the sequential and temporal organization of host-acceptance behavior inBrachymeria intermedia (Chalcididae), an endoparasitoid of gypsy moth

Experienced Brachymeria intermediafemales are almost twice as likely to accept a Lymantria disparpupa as inexperienced parasitoids. The sequence of parasitoid behaviors that leads to host acceptance is highly canalized. Experienced parasitoids, however, have a higher probability of initiating host investigation and making a transition from drumming to grasping, which in turn almost invariably leads to ovipositor insertion. Experienced females found the host more quickly than inexperienced females. Host-handling time did not change with experience but was longer in females that accepted rather than rejected the host. Females exposed to gypsy moth odor but not contacting pupae behaved similarly to females that never experienced host-related stimuli.     

寄生蜂寻找隐蔽性寄主害虫的行为机制

林木蛀干类害虫具有高度的隐蔽性,是林业上的一类重要害虫,也是目前世界上最难防治的害虫类群之一.寄生蜂在与寄主长期的协同进化过程中,形成了搜索、发现和攻击寄主害虫的独特机制,能够有效地找到并寄生它们.总结了寄生性天敌寻找、发现并成功定位隐蔽性寄主害虫的行为学机制.寄生蜂可以利用来自嗅觉的化学信息物质(如寄主、寄主粪便、虫道共生菌的挥发性气味)、寄主成虫的化学通讯物质、来自视觉的植物表面色差信息、来自触觉的寄主保护物性状特征、来自寄主取食和运动所产生的介质振动信号以及来自寄主活动和代谢的红外辐射等多种途径有效地发现隐蔽性害虫的位置,从而完成寄生行为.有些寄生蜂还能综合利用来源不同的多种信息,从而提高寄主定位的可靠性和准确性.本文还对寄生蜂寻找寄主的这些线索在生物防治上可能的利用途径和前景进行了讨论.这对促进我国在该领域的研究,充分利用天敌昆虫,提高生物防治效率具有参考价值.     

Ambient temperature affects mechanosensory host location in a parasitic wasp

Certain parasitic wasps (Ichneumonidae, Pimplinae) use self-produced vibrations transmitted on plant substrate to locate their immobile concealed hosts (i.e. lepidopteran pupae). This mechanosensory mechanism, called the vibrational sounding, depends both on physical cues of the environment and physical activity of the parasitoid and is postulated to depend on ambient temperature. We analysed the influences of temperature on vibrational sounding by choice experiments using plant-stem models with hidden host mimics in the temperate species Pimpla turionellae. The results show a significant effect of temperature on host-location activity and on the success of this process. Outside an optimum range, the performance of the wasps decreased both at low and high temperatures. Below 10°C and beyond 24°C, the wasps displayed (1) substantial reduction in responsiveness, i.e. proportion of females showing ovipositor insertions, (2) reduction of quantitative activity with ovipositor insertions in the individuals, and (3) reduced precision of mechanosensory host location. Nevertheless, female wasps were able to locate their host over a surprisingly broad range of ambient temperatures which indicates that the wasps are able to compensate for temperature effects on vibrational sounding.     

Host preference and sex allocation of three hymenopteran parasitoid species (Ichneumonidae and Braconidae) of a longicorn pest, Oemona hirta (Fabr.) (Col., Cerambycidae)

The host preference and sex allocation of two ichneumond parasitoids, Xanthocryptus novozealandicus (Dalla Torre) and Campoplex sp., and one braconid parasitoid, Apsicolpus hudsoni Turner of the longicorn borer, Oemona hirta , were studied in New Zealand. The size of the borer larvae attacked by the parasitoids can be determined by measuring the gallery width of the borers. These parasitoids are idiobionts and their secondary sex ratio is female-biased. Females of these species are significantly larger than males, and the females allocate the sex of offspring depending on the size of host larvae, by laying female-producing eggs on significantly larger host larvae. Xanthocryptus novozealandicus is larger in body size and shorter in ovipositor length than Campoplex sp. and A. hudsoni , and consumes late instar larvae or prepupae in shallow galleries whereas the latter two utilize early to middle instar larvae in deep galleries. Campoplex sp. and A. hudsoni preferred host larvae in significantly larger tree twigs than X. novozealandicus .     

Vibratory stimuli in host location by parasitic wasps

Parasitic wasps use a broad spectrum of different stimuli for host location and host acceptance. Here we review the published evidence for the use of mechanical stimuli, i.e. substrate born vibrations which are invariably regarded as vibrotaxis. We propose a set of criteria to class behavioural reactions as vibrotaxis or vibrokinesis and characterize 14 studies reporting the use of host-associated vibrations by parasitoids. The studies are compared concerning (i) experimental design; (ii) characterisation of vibrational signals; and (iii) progress of the parasitoid towards the host.The recent experimental development based on new measurement techniques shows the growing body of evidence that host-associated vibrations are exploited by parasitic wasps. Nevertheless a definite proof for vibrotaxis is still lacking. To assess the exact mechanisms by which parasitoids use vibrations bioassays comparing reactions to natural and artificially generated signals are needed. Vibrotaxis as well as vibrokinesis are both helpful host location strategies for parasitoids foraging in a multimodal environment. At the community level they may lead to niche differentiation.     

Parasitism on medfly by Diachasmimorpha longicaudata (Ashmead) (Hymenoptera: Braconidae) in different guava cultivars

The parasitism efficiency of the Braconidae wasp, Diachasmimorpha longicaudata (Ashmead), was checked on four guava cultivars (Paluma, Sassaoca, Pedro Sato and Kumagai) infested with larvae of medfly, Ceratitis capitata (Wiedemann) (Diptera: Tephritidae). Five blocks of eight fruits, each with two fruits of each cultivar, were put inside C. capitata adult cages, during 2h for oviposition, and a week later, when the larvae inside guavas were developed, the fruits were exposed to parasitoids for 24h. The mean fruit weight, larvae mortality, number of pupae, percentage of medfly and parasitoid emergence were evaluated. There was not statistical difference among cultivars to weight, larvae mortality, number of pupae e emergence of medfly. The percentage of parasitism was higher in Pedro Sato cultivar (19.8%) compared with Kumagai cultivar (2.9%), but it was statistically similar to the other cultivars.     

Native parasitoids and their potential to control the invasive leafminer, Cameraria ohridella DESCH. & DIM. (Lep.: Gracillariidae)

In spite of the fact that since the end of the eighties, the horse chestnut leafminer, Cameraria ohridella, has established itself throughout Europe, native predators such as ants and birds are not attuned to this neozoic species. In contrast, several parasitic wasp species already started to exploit the invasive horse chestnut leafminer, but until now parasitation rates are quite low, mainly because of asynchrony in the lifecycles of parasitoids and host. Only the removal of leaf litter, in which pupae hibernate, is at the moment a strategy to reduce the infestation level in the next year. Unfortunately, not only hibernating horse chestnut leafminers but also parasitoids are removed, and important resources for biocontrol are unused. In the current study, we investigated the potential efficiency of the horse chestnut leafminer parasitoid complex extracted from leaf litter in defined environments. Parasitoids were released at different densities to investigate density dependence in parasitation rates.Although seven different species were released in our experiments, only Pnigalio agraules turned out to be responsible for biocontrol of C. ohridella. We recorded parasitation rates of up to 35%. Overall, parasitation rates were independent of the leafminer density but increased fourfold if ten times more parasitoid individuals were released. Unfortunately, none of the parasitoid species could be established in the experimental units in the long run. Results are compared to other parasitoid-leafminer systems, and promotion of horse chestnut leafminer parasitoids to support natural selection and biological control of the horse chestnut leafminer is discussed.