Entomopathogens and Parasitoids Allied in Biocontrol: A Systematic Review.

Biological pest control is an environmentally friendly alternative to synthetic pesticides, using organisms such as viruses, bacteria, fungi, and parasitoids. However, efficacy is variable and combining different biocontrol agents could improve success rates. We conducted a systematic review of studies combining a parasitoid with an entomopathogenic microorganism, the first of its kind. We searched in Web of Science and extracted data from 49 publications matching the pre-defined inclusion criteria. Combinations of 36 hymenopteran parasitoids with 17 entomopathogenic microorganisms used to control 31 target pests were found. Trichogramma pretiosum and Encarsia formosa were the most frequently studied parasitoids, while Beauveria bassiana, Metarhizium anisopliae, Lecanicillium muscarium, Bacillus thuringiensis var. kurstaki, the Spodoptera exigua multiple nucleopolyhedrovirus, and the Spodoptera frugiperda multiple nucleopolyhedrovirus were the main microbial agents assessed. Out of 49 parasitoid-microorganism combinations assessed in the laboratory experiments, thirty-eight were reported as compatible and six as incompatible. Timing and dosage of biopesticides played a crucial role, with later application and appropriate dosage minimizing adverse effects on parasitoid development. More research is needed to assess compatibility and efficacy under real-world conditions. Our review provides valuable insights for researchers and practitioners to optimize the combined use of micro- and macroorganisms for effective pest control.

Assessing potential hybridization between a hypothetical gene drive-modified Drosophila suzukii and nontarget Drosophila species.

Genetically engineered gene drives (geGD) are potentially powerful tools for suppressing or even eradicating populations of pest insects. Before living geGD insects can be released into the environment, they must pass an environmental risk assessment to ensure that their release will not cause unacceptable harm to non-targeted entities of the environment. A key research question concerns the likelihood that nontarget species will acquire the functional GD elements; such acquisition could lead to reduced abundance or loss of those species and to a disruption of the ecosystem services they provide. The main route for gene flow is through hybridization between the geGD insect strain and closely related species that co-occur in the area of release and its expected dispersal. Using the invasive spotted-wing drosophila, Drosophila suzukii, as a case study, we provide a generally applicable strategy on how a combination of interspecific hybridization experiments, behavioral observations, and molecular genetic analyses can be used to assess the potential for hybridization.

Few indirect effects of baculovirus on parasitoids demonstrate high compatibility of biocontrol methods against Tuta absoluta.

BACKGROUND: Combining different biocontrol agents, particularly micro- and macroorganisms, can contribute to new and sustainable pest control approaches. Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is one of the most destructive pests of solanaceous crops. An emerging management strategy consists of biological control using microbial insecticides such as baculoviruses, but with limited efficacy. Thanks to their high target specificity, baculoviruses can be used simultaneously with natural enemies such as parasitoids for improved control of T. absoluta. However, potential indirect nontarget effects of baculoviruses on parasitoids can result from overlapping resource requirements. We assessed whether ovipositing parasitoid females discriminated against virus-treated hosts and examined the outcome of within-host competition between the hymenopteran parasitoids Necremnus tutae (Reuter) (Eulophidae) and Dolichogenidea gelechiidivoris Marsch (Braconidae), and the Phthorimaea operculella granulovirus (PhopGV, Baculoviridae) that infects T. absoluta larvae. RESULTS: Female D. gelechiidivoris discriminated against virus-treated hosts, whereas N. tutae did not. We found few indirect virus-related effects depending on the species, the sex, and the time of virus treatment. Effects were ambivalent for D. gelechiidivoris offspring and ranged from increased male longevity when infection occurred before parasitization to reduced emergence and male longevity when infection occurred after parasitization. N. tutae offspring showed a longer development time and shorter male longevity when they developed in virus-treated hosts. CONCLUSION: The virus had a low impact on parasitoid offspring. In rare cases, adverse effects were detected; however, the low magnitude of these effects is unlikely to reduce the fitness of parasitoid offspring, therefore both parasitoids seem compatible with the baculovirus for control of T. absoluta. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Overwintering of two pupal parasitoids of Drosophila under natural conditions.

The overwintering capacity of biocontrol agents is of fundamental relevance for biological control of pests in temperate regions. In this study we tested the cold tolerance of the indigenous Drosophila pupal parasitoids Pachycrepoideus vindemmiae and Trichopria drosophilae at constant low temperature in the laboratory and exposed different preimaginal parasitoid stages in the field during winter. We evaluated whether semi-natural habitats promote overwintering via more favorable microclimatic conditions as well as higher host availability compared to orchards. Further, we studied the parasitoids' phenology in a semi-field experiment during autumn. We found that P. vindemmiae larvae and pupae were most cold tolerant under laboratory and field conditions, while all preimaginal stages of T. drosophilae displayed similar cold tolerance. Semi-natural habitats buffered temperature extremes, yet overwintering survival was not enhanced compared to orchards. Suitable overwintering hosts were present in all habitats at times when parasitoids were still active parasitizing. These results demonstrate that P. vindemmiae overwinters most likely as larva or pupa and that T. drosophilae can overwinter in a preimaginal life stage. Further, we provide evidence that both parasitoids can overwinter in a wide range of habitats and that the availability of hosts for overwintering is unlikely a limiting factor for the parasitoids during fall.

Predation on Drosophila suzukii within Hedges in the Agricultural Landscape.

The invasive Drosophila suzukii feeds and reproduces on various cultivated and wild fruits and moves between agricultural and semi-natural habitats. Hedges in agricultural landscapes play a vital role in the population development of D. suzukii, but also harbor a diverse community of natural enemies. We investigated predation by repeatedly exposing cohorts of D. suzukii pupae between June and October in dry and humid hedges at five different locations in Switzerland. We sampled predator communities and analyzed their gut content for the presence of D. suzukii DNA based on the COI marker. On average, 44% of the exposed pupae were predated. Predation was higher in dry than humid hedges, but did not differ significantly between pupae exposed on the ground or on branches and among sampling periods. Earwigs, spiders, and ants were the dominant predators. Predator communities did not vary significantly between hedge types or sampling periods. DNA of D. suzukii was detected in 3.4% of the earwigs, 1.8% of the spiders, and in one predatory bug (1.6%). While the molecular gut content analysis detected only a small proportion of predators that had fed on D. suzukii, overall predation seemed sufficient to reduce D. suzukii populations, in particular in hedges that provide few host fruit resources.

Host searching and host preference of resident pupal parasitoids of Drosophila suzukii in the invaded regions.

BACKGROUND: In its invaded regions, Drosophila suzukii (Matsumura) is a novel host for the community of resident parasitoids of Drosophila. To attain a high parasitization rate on the novel host, the parasitoids have to locate it and accept it in the presence of other Drosophila hosts. We conducted a laboratory choice experiment and a semifield trial to investigate host searching and host preference of the three pupal parasitoid species Trichopria drosophilae (Perkins), Pachycrepoideus vindemmiae (Rondani) and Spalangia erythromera Förster. RESULTS: All three parasitoid species preferred D. suzukii over two common native hosts in the choice experiment. In field cages, most parasitoid offspring emerged from D. suzukii hosts. While P. vindemmiae mainly parasitized hosts in the foliage, most T. drosophilae offspring emerged from pupae presented on the ground. CONCLUSIONS: Both P. vindemmiae and T. drosophilae have the potential to find and parasitize D. suzukii in the field. If released early in the season, possible nontarget effects on native Drosophila should be minimal.

Cold tolerance of the drosophila pupal parasitoid Trichopria drosophilae.

Trichopria drosophilae (Perkins) (Hymenoptera: Diapriidae) is a pupal parasitoid of drosophila flies recorded from several parts of the world. It is currently considered for augmentative biological programs to control the severe agricultural pest Drosophila suzukii (Matsumura) (Diptera: Drosophilidae). Since D. suzukii has invaded regions that experience zero and sub-zero winter temperatures, cold tolerance of the parasitoid is an important aspect to consider. We characterized low temperature tolerance and overwintering capacity of a colony of T. drosophilae collected in Northern Switzerland. We used copper-constantan thermocouples to determine the supercooling point and pre-freeze mortality. Moreover, we subjected honey-fed and unfed adult T. drosophilae as well as developing stages within their drosophila host to short- and long-term acclimation conditions and assessed the duration of their survival at low temperatures. Finally, we exposed adult and sub-adult stages to winter conditions in a semi-field experiment and evaluated their survival. We found that T. drosophilae is chill susceptible like D. suzukii, but adults froze and survived at colder temperatures than those reported for D. suzukii. Adult parasitoids could tolerate several days of exposure to sub-zero temperatures and could reproduce afterwards, whereas sub-adult stages could survive longer periods under these conditions. The provision of honey and water enhanced the survival of adults and long-term acclimation led to longer survival in all stages. The semi-field experiment supported the results of the laboratory tests. Based on these results we suggest that in Central Europe, T. drosophilae survives winters mainly in developing stages but adults are likely able to tolerate short periods of low spring temperatures.

Multiscale Determinants Drive Parasitization of Drosophilidae by Hymenopteran Parasitoids in Agricultural Landscapes.

(1) The management of agricultural landscapes for pest suppression requires a thorough understanding of multiple determinants controlling their presence. We investigated the ecological preferences of indigenous parasitoids and their drosophilid hosts to understand the role of native parasitoids as biological control agents of the invasive frugivorous Drosophila suzukii. (2) Using data from an extensive field survey across different habitat types we analyzed the influence of abiotic and biotic factors on parasitoid and drosophilid communities at multiscale levels. (3) Eight parasitoid and 27 drosophilid species were identified. Thirty-four percent variation in drosophilid communities was explained by factors at the landscape scale, and 52% of significant variation of parasitoids by local distribution of three drosophilid species, mainly collected in woodland. Parasitoid communities were significantly influenced by microhabitat type (ground versus canopy) rather than habitat type. All parasitoids except Pachycrepoideus vindemmiae preferred the ground microhabitat. All parasitoids, with the exception of Trichopria drosophilae and Spalangia erythromera, displayed significant preferences among the drosophilid species used in the baited traps. (4) Since they can tolerate a broad range of habitat factors, altogether pupal parasitoids investigated in this study could play a role in biological control programs to suppress D. suzukii, but non-target effects have to be regarded.

Influence of the Rearing Host on Biological Parameters of Trichopria drosophilae, a Potential Biological Control Agent of Drosophila suzukii.

Trichopria drosophilae is a pupal parasitoid that can develop in a large number of drosophilid host species including the invasive pest Drosophila suzukii, and is considered a biological control agent. We investigated the influence of the rearing host on the preference and performance of the parasitoid, using two different strains of T. drosophilae, reared on D. melanogaster or D. suzukii for approximately 30 generations. Host switching was employed to assess the impact of host adaptation on T. drosophilae performance. In a no-choice experimental setup, T. drosophilae produced more and larger offspring on the D. suzukii host. When given a choice, T. drosophilae showed a preference towards D. suzukii, and an increased female ratio on this host compared to D. melanogaster and D. immigrans. The preference was independent from the rearing host and was confirmed in behavioral assays. However, the preference towards D. suzukii increased further after a host switch from D. melanogaster to D. suzukii in just one generation. Our data indicate that rearing T. drosophilae for several years on D. melanogaster does not compromise its performance on D. suzukii in the laboratory. However, producing a final generation on D. suzukii prior to release could increase its efficacy towards the pest.

Seasonal and regional presence of hymenopteran parasitoids of Drosophila in Switzerland and their ability to parasitize the invasive Drosophila suzukii.

Since its introduction into Europe the invasive Drosophila suzukii has established and spread widely, thereby entering habitats populated by native Drosophila species and their natural enemies. The highly prolific D. suzukii will likely interact with these species as a competitor, host or prey. To investigate potential interactions of D. suzukii with parasitoids, a field survey was conducted across several fruit-growing regions in Switzerland in two consecutive years. Eight species of hymenopteran parasitoids were collected using D. melanogaster as sentinel hosts in field-traps. Parasitoid capture was much higher in 2015 than in 2014 and varied among regions, time of the growing season, and habitat type. Laboratory no-choice assays with the field-collected species demonstrated that the larval parasitoids Asobara tabida, Leptopilina boulardi, and L. heterotoma could not use D. suzukii for reproduction, although the latter two reduced the number of emerging D. suzukii. In contrast, the pupal parasitoids Pachycrepoideus vindemmiae, Trichopria drosophilae, Vrestovia fidenas and Spalangia erythromera all developed with D. suzukii as hosts. Regional differences between strains were generally not evident, with the exception of two T. drosophilae strains that differed in parasitization rate. Thus, native parasitoids may interact with D. suzukii and should be regarded when implementing pest control measures.

Identification of a Grain Beetle Macrolide Pheromone and Its Synthesis by Ring-Closing Metathesis Using a Terminal Alkyne.

A major C18-macrolide was found during analysis of the frass of the storage beetle Oryzaephilus surinamensis to be (9Z,12Z,15R)-octadeca-9,12-dien-15-olide (10, cucujolide XI). The synthesis used ring-closing alkyne metathesis as a key step. The highly active 2,4,6-trimethylbenzylidyne molybdenum complex [MesCMo{OC(CF3)2Me}3] (12) allowed the use of a terminal alkyne and afforded the product in excellent yield. Bioassays proved the activity of the R-enantiomer 10 in the aggregation of the beetle. Cucujolide XI is the first macrolide pheromone oxidized at the ω-4 position.

Identification and synthesis of macrolide pheromones of the grain beetle Oryzaephilus surinamensis and the frog Spinomantis aglavei.

Macrolide lactones, the so called cucujolides derived from unsaturated fatty acids, are aggregation pheromones of cucujid grain beetles. Thirty years ago, Oehlschlarger et al. showed that (3Z,6Z)-dodeca-3,6-dien-11-olide (4) and the respective 12-olide (7) attract the sawtoothed grain beetle Oryzaephilus surinamensis, whereas (5Z,8Z,13R)-tetradeca-5,8-dien-13-olide (5) increases the response synergistically. The frass of this beetle is attractive for its parasitoid Cephalonomia tarsalis, which potentially can be used for pest control. A GC/MS analysis of attractive frass showed the presence of 5, together with an unknown isomer. Cucujolide V was tentatively identified also in the femoral glands, pheromone-releasing structures, of the Madagascan mantelline frog Spinomantis aglavei. Therefore, a new route to synthesize doubly unsaturated macrolides allowing the flexible attachment of the side chain was developed. A straightforward method to obtain Z configured macrolides involves ring-closing alkyne metathesis (RCAM) followed by Lindlar-catalyzed hydrogenation. This methodology was extended to homoconjugated diene macrolides by using RCAM after introduction of one Z configured double bond in the precursor by Wittig reaction. A tungsten benzylidyne complex was used as the catalyst in the RCAM reaction, which afforded the products in high yield at room temperature. With the synthetic material at hand, the unknown isomer was identified as the new natural product (5Z,8Z,12R)-tetradeca-5,8-dien-12-olide, cucujolide X (8). Furthermore, the route also allowed the synthesis of cucujolide V in good yield. The natural products were identified by the synthesis of enantiomerically pure or enriched material and gas chromatography on chiral phases. The new macrolide (R)-8 proved to be biologically active, attracting female O. surinamensis, but no males. The synthetic material allowed the identification of (R)-5 in both the beetle and the frog.

A host-plant-derived volatile blend to attract the apple blossom weevil Anthonomus pomorum - the essential volatiles include a repellent constituent.

BACKGROUND: Plant volatiles are promising cues for trapping pest insects. This study started with a recently identified complex blend released by prebloom apple trees and aimed to reduce the number of compounds in the blend while maintaining the attraction of the target pest, the apple blossom weevil Anthonomus pomorum. An evaluation was made to determine whether attraction to plant volatiles is a general feature in this species. RESULTS: Laboratory-based bioassays with field-collected weevils demonstrated repellency by volatiles from the non-host walnut, indicating that preference for plant odours is not a general feature in this species. By a subtractive bioassay approach, the original number of compounds in the apple-plant-released blend was stepwise reduced from 12 to 6 while maintaining weevil attraction. This resulting blend was as attractive as the full blend and as a blossom-bud-carrying apple twig. It was found to be composed of two synergistically interacting constituents, of which the first containing benzenoids was behaviourally inactive, and the second comprising the remaining compounds was even repellent. CONCLUSIONS: This study enhances knowledge of the interaction of behaviourally effective constituents in complex odour blends and contributes to the development of an efficient monitoring system involving plant volatiles for the apple blossom weevil.

Mate finding in the parasitic wasp Cephalonomia tarsalis (Ashmead): more than one way to a female's heart.

The parasitic wasp, Cephalonomia tarsalis (Hymenoptera: Bethylidae), parasitizes larvae of the sawtoothed grain beetle, Oryzaephilus surinamensis (L.) (Coleoptera: Silvanidae), and is used for biological control of this worldwide pest of stored grain. To study the hypothesis that C. tarsalis not only mates at its natal patch but also uses olfactory cues to find mating partners elsewhere, we investigated semiochemical use by male C. tarsalis. Olfactometer experiments revealed that male C. tarsalis are arrested by odors emanating from the cocoons of conspecifics, from young unmated females, and from feces of the host. Dodecanal, which was identified from extracts of filter paper contaminated by young females, had an arresting effect on males but not on females and was, therefore, considered as a sex pheromone. These findings indicate that C. tarsalis is a species with partial local mate competition. Males mate with females: 1) at the emergence site following location of females by sex pheromones from their cocoons, and 2) after dispersal from the natal patch following location of females directly by dodecanal and indirectly by unidentified sexual kairomones from host feces.

Olfactory host finding, intermediate memory and its potential ecological adaptation in Nasonia vitripennis.

Associative learning of host-associated chemical cues was studied in Nasonia vitripennis, a parasitoid of fly pupae in nests of hole-nesting birds. When females encountered a fly pupa and performed one sequence of host recognition behaviour including drilling the ovipositor into the host in the presence of the artificial odour furfurylheptanoate (FFH), they were afterwards arrested by FFH in olfactometer experiments. The response vanished after 4 days and could be blocked after 3 days by feeding wasps with ethacrynic acid prior and after the training. This indicates the formation of an intermediate form of memory by one host experience in N. vitripennis. Interestingly, the trained wasps avoided odours that were not present during the host encounter, although naive wasps did not react to these odours. This unique behaviour probably causes wasps to focus during host searching on those chemical cues they have experienced in the host environment. Studies in nests of hole-nesting birds revealed that about 30% of all nests contained only one fly pupa, and laboratory studies showed that N. vitripennis females are able to parasitise around 100 fly pupae in their life. It is discussed that under these conditions, the formation of a non-permanent intermediate memory for host-associated odours after one host encounter is adaptive to avoid costs involved with formation and maintenance of memory for misleading cues. The demonstration of associative olfactory learning in N. vitripennis, the first parasitoid species with sequenced genome, opens the gate to study molecular mechanisms of memory formation and its ecological adaptation in parasitoids.

Protein synthesis-dependent long-term memory induced by one single associative training trial in the parasitic wasp Lariophagus distinguendus.

Protein synthesis-dependent long-term memory in Apis mellifera and Drosophila melanogaster is formed after multiple trainings that are spaced in time. The parasitic wasp Lariophagus distinguendus remarkably differs from these species. It significantly responds to the artificial odor furfurylheptanoate (FFH) in olfactometer experiments, when this odor was presented during one single training trial, consisting of one sequence of host recognition behavior on a wheat grain infested by its hosts. Feeding wasps with actinomycin D erases the learned response 24 h after the training, demonstrating that protein synthesis-dependent long-term memory has been formed in L. distinguendus already after one single training.

Identification and biosynthesis of an aggregation pheromone of the storage mite Chortoglyphus arcuatus.

In an effort to identify new pheromones from mites, the headspace of undisturbed colonies of the storage mite Chortoglyphus arcuatus was analyzed by GC-MS by use of a closed-loop stripping apparatus (CLSA) or solid-phase microextraction (SPME). The major compound emitted from the mites is (4R,6R,8R)-4,6,8-trimethyldecan-2-one (4R,6R,8R-8). The structure was elucidated by analysis of the mass spectrum, synthesis of authentic samples, and gas chromatography on a chiral phase. Bioassays show that this compound, for which we propose the trivial name chortolure, is an aggregation pheromone for both sexes of this species. Several related compounds are released in smaller amounts by the mites. The alarm pheromones of these mites, neral and geranial, can only be found in total extracts of the mites, in which 8 occurs only in minute amounts. The method of sampling is therefore crucial for pheromone identification. Feeding experiments with deuterated propionate showed that chortolure is a polyketide, formed by successive addition of four propionate units to an acetate starter.