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The egg parasitoid Trichogramma spp. (Hymenoptera: Trichogrammatidae) is a widely used biocontrol agent against lepidopteran pests. Historically, Trichogramma were deployed either by plane or by using cardboard cards on which parasitized eggs are glued and manually installed at sites. Plane deployment is costly and card installation is time consuming, but the use of Trichogramma has been shown to be efficient against several pests. In 2016 and 2017, a research project investigated the potential use of unmanned aerial system for distributing Trichogramma as biocontrol agents against two major pests: an agricultural pest of maize, the European corn borer, Ostrinia nubilalis (Hübner) (Lepidoptera: Crambidae), and a forest pest, the eastern spruce budworm, Choristoneura fumiferana (Clemens) (Lepidoptera:Tortricidae). Exposure duration of parasitized eggs to field conditions (temperature, predation, etc.) in maize fields influenced the Trichogramma's emergence rate, suggesting that timing of parasitoid releases with their emergence is essential. Although parasitism of naturally occurring eggs in maize fields could not be compared due to the low density of the European corn borer, parasitism of sentinel eggs by Trichogramma was more prominent in plots with unmanned aircraft systems (UAS)-releases compared to control plots. For spruce budworm, treatment with Trichogramma increased egg parasitism and there was no difference between the deployment by UAS and by Trichocards. We discuss these results in the context of pest biology and management. We also discuss the advantages and shortcomings of both methods and offer insights into where future work might go to further leverage the use of UAS in managing these important pests.

Animal mitochondrial genomes usually exhibit conserved gene arrangement across major lineages, while those in the Hymenoptera are known to possess frequent rearrangements, as are those of several other orders of insects. Here, we sequenced two complete mitochondrial genomes of Trichogramma japonicum and Trichogramma ostriniae (Hymenoptera: Chalcidoidea: Trichogrammatidae). In total, 37 mitochondrial genes were identified in both species. The same gene arrangement pattern was found in the two species, with extensive gene rearrangement compared with the ancestral insect mitochondrial genome. Most tRNA genes and all protein-coding genes were encoded on the minority strand. In total, 15 tRNA genes and seven protein-coding genes were rearranged. The rearrangements of cox1 and nad2 as well as most tRNA genes were novel. Phylogenetic analysis based on nucleotide sequences of protein-coding genes and on gene arrangement patterns produced identical topologies that support the relationship of (Agaonidae + Pteromalidae) + Trichogrammatidae in Chalcidoidea. CREx analysis revealed eight rearrangement operations occurred from presumed ancestral gene order of Chalcidoidea to form the derived gene order of Trichogramma . Our study shows that gene rearrangement information in Chalcidoidea can potentially contribute to the phylogeny of Chalcidoidea when more mitochondrial genome sequences are available.

The identification of Trichogramma (Hymenoptera: Trichogrammatidae) species is problematic due to their small size and lack of distinct morphological characters. In this study, we combined morphological characters of the male genitalia and molecular methods using the mitochondrial cytochrome oxidase I (COI) gene as a molecular marker to identify eight species from 16 geographic populations: T. evanescens Westwood, T. cacoeciae Marchal, T. ostriniae Pang et Chen, T. chilonis Ishii, T. japonicum Ashmead, T. brassicae Bezdenko, T. bilingensis He et Pang, and T. dendrolimi Matsumura. We developed a polymerase chain reaction–restriction fragment length polymorphism (PCR–RFLP) method targeting the mitochondrial COI to distinguish the eight Trichogramma species using three restriction enzymes. We further analyzed 109 COI fragments from 18 Trichogramma species and found that the PCR–RFLP method could distinguish both intra- and inter-specific genetic variation among most of the species using four additional restriction enzymes.

Chalcidoidea (Hymenoptera) are extremely diverse with more than 23,000 species described and over 500,000 species estimated to exist. This is the first comprehensive phylogenetic analysis of the superfamily based on a molecular analysis of 18S and 28S ribosomal gene regions for 19 families, 72 subfamilies, 343 genera and 649 species. The 56 outgroups are comprised of Ceraphronoidea and most proctotrupomorph families, including Mymarommatidae. Data alignment and the impact of ambiguous regions are explored using a secondary structure analysis and automated (MAFFT) alignments of the core and pairing regions and regions of ambiguous alignment. Both likelihood and parsimony approaches are used to analyze the data. Overall there is no impact of alignment method, and few but substantial differences between likelihood and parsimony approaches. Monophyly of Chalcidoidea and a sister group relationship between Mymaridae and the remaining Chalcidoidea is strongly supported in all analyses. Either Mymarommatoidea or Diaprioidea are the sister group of Chalcidoidea depending on the analysis. Likelihood analyses place Rotoitidae as the sister group of the remaining Chalcidoidea after Mymaridae, whereas parsimony nests them within Chalcidoidea. Some traditional family groups are supported as monophyletic (Agaonidae, Eucharitidae, Encyrtidae, Eulophidae, Leucospidae, Mymaridae, Ormyridae, Signiphoridae, Tanaostigmatidae and Trichogrammatidae). Several other families are paraphyletic (Perilampidae) or polyphyletic (Aphelinidae, Chalcididae, Eupelmidae, Eurytomidae, Pteromalidae, Tetracampidae and Torymidae). Evolutionary scenarios discussed for Chalcidoidea include the evolution of phytophagy, egg parasitism, sternorrhynchan parasitism, hypermetamorphic development and heteronomy.

Temperature has a profound effect on performance and behavior of egg parasitoids. Egg parasitoids are a well-known alternative for the control of lepidopterous pests. Selected life history parameters of Trichogramma euproctidis (Girault) (Hymenoptera: Trichogrammatidae), an established egg parasitoid species in Khuzestan-Southwest Iran, were appraised at eight constant temperatures (22.5, 25, 27.5, 30, 32.5, 35, 37.5, and 40°C) using Ephestia kuehniella Zeller (Lepidoptera: Pyralidae) eggs as the host. We found significant effects of temperature on the number of parasitized eggs, development time, sex ratio, progeny's longevity, and fecundity. T. euproctidis developed on E. kuehniella eggs at all temperatures tested, but performed best at 32.5°C. At this temperature, they parasitized the most eggs, produced the most female progeny, and had high rates of survival. Our findings revealed that temperature significantly affected the longevity of female progeny and fecundity of T. euproctidis. A life table analysis confirmed that temperature resulted in optimal effects on T. euproctidis life history. Net reproductive rate (R0) of T. euproctidis was different among the temperatures tested. The intrinsic rate of increase (r) was positively correlated with temperature from 22.5 to 32.5°C and then decreased from 35 to 40°C. Generation time (T) and doubling time (DT) decreased as temperature increased from 22.5 to 37.5°C and then increased at 40°C. These data suggest that this strain of T. euproctidis is adapted to high temperatures and harsh environmental conditions and has the potential to be used in integrated management programs in Southwest Iran.

Trichogramma spp. wasps are egg parasitoids with a long history of mass rearing for augmentation biocontrol programs in field crop and orchard landscapes. Supplementary nutrition can improve the longevity, fecundity, and biocontrol efficacy of parasitoids. To improve the production efficiency and parasitism performance of Trichogramma dendrolimi Matsumura (Hymenoptera: Trichogrammatidae), the present study screened and examined the potential supplementary nutrients for this biological control agent. Dietary supplementation with a 10% sucrose solution significantly increased wasp longevity and parasitism potential of T. dendrolimi on host eggs, but provision of pollen did not provide additional benefits. Laboratory and greenhouse cage tests demonstrated that wasp access to soybean aphid Aphis glycines Matsumura (Hemiptera: Aphididae) honeydew, comprised primarily of melezitose and trehalose, improved T. dendrolimi longevity and parasitism. In conclusion, provision of a 10% sucrose solution to adult wasps will enhance the mass-rearing efficiency of T. dendrolimi; furthermore, field release of T. dendrolimi by plant vectors bearing honeydew-producing aphids holds promise for improving the biocontrol efficacy of T. dendrolimi. Graphical Abstract

Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) is a polyphagous pest with a wide geographic distribution. This pest first arrived in Brazil in 2013, and since then studies on possible control methods for it have been necessary. A possible method for the control of H . armigera is using the egg parasitoid Trichogramma pretiosum Riley (Hymenoptera: Trichogrammatidae). Therefore, the objective of this study was to evaluate the performance of T . pretiosum on H . armigera eggs, which are known to represent suitable hosts for the development of this parasitoid species in the laboratory. Parasitism and emergence rates and the duration of the egg-to-adult period of T . pretiosum were investigated following 24- and 48-h exposures of this parasitoid to H . armigera and Corcyra cephalonica (Stainton) (Lepidoptera: Pyralidae) eggs. The longevity of offspring after the 24-h exposure was studied, as well as the frequency of parasitism and emergence, host preference, and the emergence of offspring from eggs of different ages or oviposited by lepidopterans on different days. Parasitism was 14.4 and 34.9% more frequent on C . cephalonica than on H . armigera after 24 and 48 h of exposure, respectively. In C . cephalonica , parasitism was 27.2% higher after 48 h. Parasitism was more frequent on C . cephalonica eggs collected on the second day of oviposition (76.2%), and on H . armigera on the third day of oviposition (71.1%). Parasitism frequency was lower on 2-day-old C . cephalonica eggs (63.3%) and on 3-day-old H . armigera eggs (41.3%). When tested with a chance of choice between hosts, T . pretiosum preferred H . armigera , while in the test with no chance of choice there was no difference in preference. Thus, T . pretiosum may be considered a tool for the Integrated Pest Management (IPM) of H . armigera .

Selecting the candidate agents or species is a fundamental step in developing effective biological control programs. However, no attempts have been made to evaluate the efficacy of Trichogramma Westwood (Hymenoptera: Trichogrammatidae) strains against the Asian corn borer, Ostrinia furnacalis Guenée (Lepidoptera: Crambidae) in Myanmar. In this study, Trichogramma strains were investigated through a series of glass tube bioassays and under a semifield condition. Ostrinia furnacalis or Corcyra cephalonica egg preferences were compared among six strains of Trichogramma ostriniae, three of Trichogramma chilonis, and four of Trichogramma dendrolimi using a choice-test assay design. Significant differences were observed at the inter- and intraspecific levels. Four strains of T. ostriniae and two of T. dendrolimi showed a strong preference for O. furnacalis eggs, while two strains of T. chilonis and one T. dendrolimi strain preferred Corcyra cephalonica Stainton (Lepidoptera: Pyralidae) eggs. The remaining strains showed no preference. Eleven strains were examined on O. furnacalis eggs only, wherein parasitism, sex ratio, and total progeny per female were highest for two strains of T. ostriniae (respectively: 67.6 ± 3.0%, 82.7 ± 2.3%, 49.6 ± 2.8, and 67.6 ± 3.6%, 90.0 ± 2.4%, 42.7 ± 2.6), and one strain of T. dendrolimi (65.2 ± 3.2%, 84.7 ± 2.6%, 46.3 ± 2.8). Parasitism and searching capacity were evaluated for the three, best performing strains in cages containing maize plants. One T. ostriniae from Southern Shan State, Myanmar parasitized ∼81% of egg masses, and should be considered a candidate biological control agent against O. furnacalis in Myanmar. Graphical Abstract

The structure of compound eyes in arthropods has been the subject of many studies, revealing important biological principles. Until recently, these studies were constrained by the two-dimensional nature of available ultrastructural data. By taking advantage of the novel three-dimensional ultrastructural dataset obtained using volume electron microscopy, we present the first cellular-level reconstruction of the whole compound eye of an insect, the miniaturized parasitoid wasp Megaphragma viggianii . The compound eye of the female M. viggianii consists of 29 ommatidia and contains 478 cells. Despite the almost anucleate brain, all cells of the compound eye contain nuclei. As in larger insects, the dorsal rim area of the eye in M. viggianii contains ommatidia that are believed to be specialized in polarized light detection as reflected in their corneal and retinal morphology. We report the presence of three ‘ectopic’ photoreceptors. Our results offer new insights into the miniaturization of compound eyes and scaling of sensory organs in general.

Background The cotton leafhopper, Amrasca biguttula Ishida (Hemiptera: Cicadellidae), is a significant pest that feeds on various host plants, particularly cotton, resulting in substantial annual economic losses. Trichogramma chilonis Ishii (Hymenoptera: Trichogrammatidae) is an effective parasitoid that targets the eggs of various pests preventing them from hatching into harmful larvae. Its ability to be mass-reared makes it a popular choice as biological control agent in integrated pest management programs worldwide. The presence of the bacterium Wolbachia , an intracellular symbiont found in arthropods, has a profound impact on the biology of their hosts. This research examined Wolbachia strain diversity in both species using seven genes: wsp , 16S rRNA , and five housekeeping MLST genes. Additionally, the molecular identification of A. biguttula and its egg parasitoid, T . chilonis was also accomplished through the use of the cytochrome c oxidase subunit I ( COI ) gene. Results Phylogenetic exploration based on 16S rRNA , wsp , and multilocus sequence typing (MLST) revealed that A. biguttula anchorages two new Wolbachia strains along with another Wolbachia strain in its egg parasitoid, T. chilonis from supergroup (B) We enumerated Wolbachia strains in A. biguttula from both study locations in Pakistan, identifying distinct strains w Abig1 and w Abig2 for A. biguttula and w TchiB for T. chilonis . Wolbachia amplification in A. biguttula showed infection rates of 29% to 66% across seven genes, with the highest rates in Bahawalpur, followed by Faisalabad. Ten adult samples of the egg parasitoid T. chilonis from each locality were also tested, revealing infection rates of 50% to 90%, again highest in Bahawalpur. Conclusion This is the first report of Wolbachia in A. biguttula populations, detecting high Wolbachia prevalence and suggesting vertical transmission. Although no additional strains were identified beyond those mentioned in the present research, further research should investigate the potential for additional strains within these populations. Wolbachia bacteria are important for pest management as they suppress pest populations, reduce disease transmission in vectors and provide pathogen resistance to hosts, while potentially increasing pest susceptibility to insecticides. As a sustainable and species-specific alternative to chemical pesticides, Wolbachia offers environmentally friendly control strategies for pests like planthoppers.