Dipteran endoparasitoid Exorista bombycis utilizes antihemocyte components against host defense of silkworm Bombyx mori.

Dipteran endoparasitoids avoid host immune response; however, antidefense components from the Dipterans are unknown. Infestation of commercial silkworm Bombyx mori Linnaeus (Lepidoptera: Bombycidae) by endoparasitoid Exorista bombycis Louis (Diptera: Tachinidae) induced immune reactions, cytotoxicity, granulation, degranulation, and augmented release of cytotoxic marker enzyme lactate dehydrogenase (LDH), and degranulation-mediator enzyme ß-hexosaminidase in hemocytes. In this study, by reverse phase high-performance liquid chromatography, fractions of E. bombycis larval tissue protein with antihemocytic activity are separated. From the fraction, peptides of hemocyte aggregation inhibitor protein (HAIP) and pyridoxamine phosphate oxidase (PNPO) are identified by mass spectrometry. Interacting partners of HAIP and PNPO are retrieved that further enhance the virulence of the parasitoid. PNPO and HAIP genes showed a four- to seven fold increase in expression in the integument of the parasitoid larva. Together, the dipteran endoparasitoid E. bombycis exploit antihemocyte activity to inhibit host defense reactions in addition to defense evasion contemplated.

Tachinid parasitoid Exorista japonica affects the utilization of diet by changing gut microbial composition in the silkworm, Bombyx mori.

Changes in both intake and digestion of feed have been demonstrated in the host following parasitization. However, its regulatory mechanism has not been clarified. In this study, silkworms and Exorista japonica were used as research objects to analyze the effect of parasitism on the midgut immune system of the silkworm. After being parasitized, the expressions of antimicrobial peptide (AMP) genes of silkworms showed a fluctuating trend of first upregulation and then downregulation, while phenoloxidase and lysozyme activities were inhibited. To study the possible impact of the downregulation of AMP genes on intestinal microorganisms, the characteristics of the intestinal microbial population of silkworms on the third day of parasitism were analyzed. The relative abundance of Firmicutes, Proteobacteria, and Bacteroidota decreased, while that of Actinobacteriota increased. The increased abundance of conditionally pathogenic bacteria Serratia and Staphylococcus might lead to a decrease in the amount of silkworm ingestion. Meanwhile, the abundance of Acinetobacter, Bacillus, Pseudomonas, and Enterobacter promotes an increase in the digestion of nutrients. This study indicated that the imbalance of intestinal microbial homeostasis caused by parasitism may affect the absorption and digestion of nutrients by the host. Collectively, our findings provided a new clue for further exploring the mechanism of nutrient transport among the host, parasitoid, and intestinal microorganisms.

Exorista sorbillans (Diptera: Tachinidae) parasitism shortens host larvae growth duration by regulating ecdysone and juvenile hormone titers in Bombyx mori (Lepidoptera: Bombycidae).

The tachinid fly, Exorista sorbillans, is a notorious ovolarviparous endoparasitoid of the silkworm, Bombyx mori, causing severe damage to silkworm cocoon industry. Silkworm larvae show typically precocious wandering behavior after being parasitized by E. sorbillans; however, the underlying molecular mechanism remains unexplored. Herein, we investigated the changes in the levels of 20-hydroxyecdysone (20E) and juvenile hormone (JH) titer, and they both increased in the hemolymph of parasitized silkworms. Furthermore, we verified the expression patterns of related genes, which showed an upregulation of 20E signaling and biosynthesis genes but a significant downregulation of ecdysone oxidase (EO), a 20E inactivation enzyme, in parasitized silkworms. In addition, related genes of the JH signaling were activated in parasitized silkworms, while related genes of the JH degradation pathway were suppressed, resulting in an increase in JH titer. Notably, the precocious wandering behavior of parasitized silkworms was partly recoverable by silencing the transcriptions of BmCYP302A1 or BmCYP307A1 genes. Our findings suggest that the developmental duration of silkworm post parasitism could be shortened by regulation of 20E and JH titers, which may help silkworm to resist the E. sorbillans infestation. These findings provide a basis for deeper insight into the interplay between silkworms and E. sorbillans and may serve as a reference for the development of a novel approach to control silkworm myiasis.

Molecular phylogeny and evolution of world Tachinidae (Diptera).

We reconstructed phylogenetic relationships within the diverse parasitoid fly family Tachinidae using four nuclear loci (7800 bp) and including an exceptionally large sample of more than 500 taxa from around the world. The position of the earthworm-parasitizing Polleniinae (Calliphoridae s.l.) as sister to Tachinidae is strongly supported. Our analyses recovered each of the four tachinid subfamilies and most recognized tribes, with some important exceptions in the Dexiinae and Tachininae. Most notably, the tachinine tribes Macquartiini and Myiophasiini form a clade sister to all other Tachinidae, and a clade of Palpostomatini is reconstructed as sister to Dexiinae + Phasiinae. Although most nodes are well-supported, relationships within several lineages that appear to have undergone rapid episodes of diversification (basal Dexiinae and Tachininae, Blondeliini) were poorly resolved. Reconstructions of host use evolution are equivocal, but generally support the hypothesis that the ancestral host of tachinids was a beetle and that subsequent host shifts to caterpillars may coincide with accelerated diversification. Evolutionary reconstructions of reproductive strategy using alternative methods were incongruent, however it is most likely that ancestral tachinids possessed unincubated, thick shelled eggs from which incubated eggs evolved repeatedly, potentially expanding available host niches. These results provide a broad foundation for understanding the phylogeny and evolution of this important family of parasitoid insects. We hope it will serve as a framework to be used in concert with morphology and other sources of evidence to revise the higher taxonomic classification of Tachinidae and further explore their evolutionary history and diversification.

Novel host immune evasion strategy of the endoparasitoid Drino inconspicuoides.

The tachinid fly Drino inconspicuoides (Diptera: Tachinidae) is an ovolarviparous endoparasitoid whose larvae develop in the host haemocoel and avoids the host immune system. In this study, we investigated the immune evasion mechanisms of this species during infestation in the host Mythimna separata (Lepidoptera: Noctuidae). We discovered a unique 'cloak' that surrounded D. inconspicuoides larvae that penetrated into the host and determined through genomic polymerase chain reaction analysis that this structure originated from the host rather than the tachinid. The 'cloak' contained both haemocytes and fat body cells from the host, with the haemocytes assembling around the larvae first and the fat body cells then covering the haemocyte layer, following which the two mixed. Living D. inconspicuoides larvae that were wrapped in the 'cloak' were not melanized whereas encapsulated dead larvae were melanized, suggesting that this structure contributes to the avoidance of host immune reactions.

Natural Enemies of the fall armyworm (Spodoptera frugiperda Smith) and comparing Neem aqueous extracts with its larvae, Gurage zone, central Ethiopia.

The fall armyworm (Spodoptera frugiperda Smith) is an invasive and polyphagous insect pest. It poses a significant threat to maize crops, uncontrolled infestation can result 100 % loss. However, natural enemies play a vital role in regulating the population of this pest. Additionally, botanical sources extracts have the potential to be effective insecticides. The objectives of the study were to investigate the natural enemies of S. frugiperda in the Gurage zone and to compare efficacy of Neem seed and leaf aqueous extracts with S. frugiperda larvae, central Ethiopia. S. frugiperda larvae and egg masses, cocoons and larvae cadavers collected from infested maze farms. From each round collection 25 healthy and inactive larvae were sampled to rear until emerging adults. Observed predator species recorded. Neem seed and leaf aqueous extracts was tested against S. frugiperda in laboratory condition. The study found a diverse range of natural enemies associated with S. frugiperda, including parasitoids, predators, and entomopathogenic fungi. Three species of parasitoids (Exorista xanthaspis, Tachina spp., and Charops annulipes) were documented in Ethiopia for the first time. Predatory insects belonging to four distinct orders: Hemiptera, Dermaptera, Coleoptera, and Mantodea also identified. In particular, various Hemipterans were observed in the maize farms infested with S. frugiperda. In terms of Neem seed and leaf aqueous extracts, they demonstrated similar mortality rates for S. frugiperda larvae after 72 h, although differences were observed at 24 and 48 h. For effective management of S. frugiperda, more research is needed to fully exploit the potential of natural enemies and botanical source insecticides.

The genome sequence of a tachinid fly, Linnaemya tessellans (Robineau-Desvoidy, 1830).

We present a genome assembly from one male Linnaemya tessellans (tachinid fly; Arthropoda; Insecta; Diptera; Tachinidae). The genome sequence is 709.9 megabases in span. Most of the assembly is scaffolded into 7 chromosomal pseudomolecules, including the X and Y sex chromosomes. The mitochondrial genome has also been assembled and is 17.24 kilobases in length.

Three new species and two newly recorded species of Tachininae from Tibet, China (Arthropoda, Insecta, Diptera, Tachinidae).

During our studying of the fauna of Tibet, China, many specimens of the subfamily Tachininae (Diptera, Tachinidae) were collected and examined. Three species are described here as new to science, Leskialatisurstylasp. nov., Trichoformosomyiacuonaensissp. nov., and Tachinajilongensissp. nov., and two species, Nemoraeajavana (Brauer & Bergenstamm, 1894) and N.echinata Mesnil, 1953, are newly recorded from Tibet. In addition to their descriptions, illustrations, and diagnoses, three identification keys are provided. The specimens in this study are kept in the Insect Collection of Shenyang Normal University, China (SYNU).

Cloak Scavenges the Reactive Oxygen Species around the Larvae of Drino inconspicuoides (Diptera: Tachinidae).

Drino inconspicuoides (Diptera: Tachinidae) is an endoparasitoid that develops inside the lepidopteran host. When the larva of D. inconspicuoides penetrates into the host, Mythimna separata (Lepidoptera: Noctuidae), the larva creates a cap-like structure, called the funnel, by using host hemocytes, forming a respiratory attachment to permit efficient respiration. A newly described cloudy and cottony structure, called the "cloak", is formed outside the funnel within 24 h of parasitism. The cloak contains the host fat body and hemocytes. In this study, we aimed to clarify the function of the cloak, which has to date remained unknown. We hypothesized that the funnel generates reactive oxygen species (ROS) through melanization, and that the cloak detoxifies them. We confirmed that the black pigments of the funnel were caused by melanization, which inevitably generates ROS that are potentially harmful to the D. inconspicuoides larva inside the funnel. The cloak showed high activities of antioxidant enzymes, including superoxide dismutase, glutathione peroxidase, and catalase. These results suggest that the cloak scavenged the ROS from the melanized funnel through the diversion of antioxidant enzymes in the fat body, thereby protecting the D. inconspicuoides larva from oxidative damage.

The genome sequence of a tachinid fly, Tachina lurida (Fabricius, 1781).

We present a genome assembly from an individual female Tachina lurida (a tachinid fly; Arthropoda; Insecta; Diptera; Tachinidae). The genome sequence is 899.2 megabases in span. Most of the assembly is scaffolded into 6 chromosomal pseudomolecules, including the X sex chromosome. The mitochondrial genome has also been assembled and is 17.3 kilobases in length. Gene annotation of this assembly on Ensembl identified 12,127 protein coding genes.

The genome sequence of a tachinid fly, Thelaira solivaga (Harris, 1780).

We present a genome assembly from an individual male Thelaira solivaga (a tachinid fly; Arthropoda; Insecta; Diptera; Tachinidae). The genome sequence is 429.3 megabases in span. Most of the assembly is scaffolded into 7 chromosomal pseudomolecules, including the X and Y sex chromosomes. The mitochondrial genome has also been assembled and is 21.09 kilobases in length.

The genome sequence of a tachinid fly, Nowickia ferox (Panzer, 1809).

We present a genome assembly from an individual female Nowickia ferox (a tachinid fly; Arthropoda; Insecta; Diptera; Tachinidae). The genome sequence is 670.7 megabases in span. Most of the assembly is scaffolded into 6 chromosomal pseudomolecules, including the X sex chromosome. The mitochondrial genome has also been assembled and is 17.19 kilobases in length. Gene annotation of this assembly on Ensembl identified 27,893 protein coding genes.

The genome sequence of a tachinid fly, Cistogaster globosa (Fabricius, 1775).

We present a genome assembly from an individual male Cistogaster globosa (a tachinid fly; Arthropoda; Insecta; Diptera; Tachinidae). The genome sequence is 837.8 megabases in span. Most of the assembly is scaffolded into 7 chromosomal pseudomolecules, including the X and Y sex chromosomes. The mitochondrial genome has also been assembled and is 16.97 kilobases in length. Gene annotation of this assembly on Ensembl identified 29,591 protein coding genes.

Parasitism by the Tachinid Parasitoid Exorista japonica Leads to Suppression of Basal Metabolism and Activation of Immune Response in the Host Bombyx mori.

The dipteran tachinid parasitoids are important biocontrol agents, and they must survive the harsh environment and rely on the resources of the host insect to complete their larval stage. We have previously demonstrated that the parasitism by the tachinid parasitoid Exoristajaponica, a pest of the silkworm, causes pupation defects in Bombyx mori. However, the underlying mechanism is not fully understood. Here, we performed transcriptome analysis of the fat body of B. mori parasitized by E. japonica. We identified 1361 differentially expressed genes, with 394 genes up-regulated and 967 genes down-regulated. The up-regulated genes were mainly associated with immune response, endocrine system and signal transduction, whereas the genes related to basal metabolism, including energy metabolism, transport and catabolism, lipid metabolism, amino acid metabolism and carbohydrate metabolism were down-regulated, indicating that the host appeared to be in poor nutritional status but active in immune response. Moreover, by time-course gene expression analysis we found that genes related to amino acid synthesis, protein degradation and lipid metabolism in B. mori at later parasitization stages were inhibited. Antimicrobial peptides including Cecropin A, Gloverin and Moricin, and an immulectin, CTL11, were induced. These results indicate that the tachinid parasitoid perturbs the basal metabolism and induces the energetically costly immunity of the host, and thus leading to incomplete larval-pupal ecdysis of the host. This study provided insights into how tachinid parasitoids modify host basal metabolism and immune response for the benefit of developing parasitoid larvae.

Effect of Tachinid Parasitoid Exorista japonica on the Larval Development and Pupation of the Host Silkworm Bombyx mori.

The Tachinidae are natural enemies of many lepidopteran and coleopteran pests of crops, forests, and fruits. However, host-tachinid parasitoid interactions have been largely unexplored. In this study, we investigated the effects of tachinids on host biological traits, using Exorista japonica, a generalist parasitoid, and the silkworm Bombyx mori, its lepidopteran host, as models. We observed that E. japonica parasitoidism did not affect silkworm larval body weight gain and cocooning rate, whereas they caused shortened duration of molting from the final instar to the pupal stage, abnormal molting from larval to pupal stages, and a subsequent decrease in host emergence rate. Moreover, a decrease in juvenile hormone (JH) titer and an increase in 20-hydroxyecdysone (20E) titer in the hemolymph of parasitized silkworms occurred. The transcription of JH and 20E responsive genes was downregulated in mature parasitized hosts, but upregulated in parasitized prepupae while Fushi tarazu factor 1 (Ftz-f1), a nuclear receptor essential in larval ecdysis, showed dramatically reduced expression in parasitized hosts at both the mature and prepupal stages. Moreover, the transcriptional levels of BmFtz-f1 and its downstream target genes encoding cuticle proteins were downregulated in epidermis of parasitized hosts. Meanwhile, the content of trehalose was decreased in the hemolymph, while chitin content in the epidermis was increased in parasitized silkworm prepupae. These data reveal that the host may fine-tune JH and 20E synthesis to shorten developmental duration to combat established E. japonica infestation, while E. japonica silences BmFtz-f1 transcription to inhibit host pupation. This discovery highlights the novel target mechanism of tachinid parasitoids and provides new clues to host/tachinid parasitoid relationships.

A tritrophic interaction at the Brazilian triple frontier: new record of parasitism on Conocephalus saltator (Sausurre, 1859) (Orthoptera, Tettigoniidae)

ABSTRACT We report for the first time a tritrophic relationship (host-parasitoid-hyperparasitoid) among Conocephalus saltator (Sausurre, 1859) (Orthoptera, Tettigoniidae), Ormia cfr. crespoi Tavares, 1965 (Diptera, Tachinidae), and Perilampus sp. (Hymenoptera, Perilampidae). Specimens of C. saltator were collected at the Parque Nacional do Iguaçu (Foz do Iguaçu, Paraná, Brazil) and reared in the laboratory, in order to detect eventual parasitoids. We collected and reared 904 katydids, with 113 of them parasitized, producing 123 fly puparia, and 18 puparia that developed to adults of Ormia cfr. crespoi. We also recorded the emergence of four hyperparasitoid wasps, Perilampus sp., from the fly puparia.

Crickets of the genus Gryllus in the United States (Orthoptera: Gryllidae: Gryllinae).

Gryllus field and wood crickets of the United States, mostly west of the Mississippi River, are reviewed and revised. We validate the following 18 Gryllus cricket names: G. armatus, G. assimilis, G. brevicaudus, G. cayensis, G. cohni, G. firmus, G. fultoni, G. integer, G. lineaticeps, G. multipulsator, G. ovisopis, G. pennsylvanicus, G. personatus, G. rubens, G. texensis, G. veletis, G. vernalis, and G. vocalis. We synonymize G. alogus under G. vocalis. We designate a lectotype for G. armatus. We describe the following 17 new Gryllus species: G. chisosensis, G. leei, G. lightfooti, G. longicercus, G. makhosica, G. montis, G. navajo, G. planeta, G. regularis, G. saxatilis, G. sotol, G. staccato, G. thinos, G. transpecos, G. veintinueve, G. veletisoides, and G. vulcanus. We present biology, distribution, and genetic analysis of all taxa and discuss their nearest relatives.

Effects of Storage at Suboptimal Temperatures on the In Vitro-Reared Parasitoid Exorista larvarum (Diptera: Tachinidae).

Exorista larvarum (L.) is a tachinid parasitoid native to the Palearctic region, known as an antagonist of lepidopterous defoliators. This species is suitable to be cultured in vitro, and yields of fecund adults, approaching those usually attained in host larvae, have been previously achieved on artificial media. Direct oviposition by E. larvarum on media has not yet been obtained, and the eggs for the in vitro rearing are routinely removed from parasitized host larvae. However, many eggs are usually laid throughout the cage by captive females and can be retrieved by placing them on artificial media. Storage at low temperatures provides a method for prolonging the development of insects and stockpile them when not needed immediately. We studied the effects of storage at 20 °C (for 5 d or until pupation) or 15 °C (for 5 d or until egg hatching) on the in vitro development of E. larvarum. Lower temperatures were excluded, because previous studies showed a strong negative impact on hatching when the eggs were stored at 5 °C or 10 °C. For the experiments, eggs were removed from hosts and placed on an artificial medium. The results suggested that it is possible to delay the development of the in vitro-reared E. larvarum, which eventually reached the adult stage, although some negative effects on fly quality (i.e., longevity and fecundity) were also observed. Because quality is not an absolute concept, all the situations tested in this study could be appropriate according to the current requirements.

A review of insect parasitoids associated with Lobesia botrana (Denis & Schiffermüller, 1775) in Italy. 1. DipteraTachinidae and HymenopteraBraconidae (Lepidoptera, Tortricidae).

This paper is aimed to summarize the information available on the parasitoid complex of the European Grapevine Moth (EGVM), Lobesia botrana (Denis & Schiffermüller, 1775) (Lepidoptera Tortricidae) in Italy. The list is the result of the consultation of a vast bibliography published in Italy for almost two hundred years, from 1828 to date. This allowed the clarification and correction of misunderstandings and mistakes on the taxonomic position of each species listed. In Italy the complex of parasitoids detected on EGVM includes approximately 90 species belonging to ten families of Hymenoptera (Braconidae, Ichneumonidae, Chalcididae, Eulophidae, Eupelmidae, Eurytomidae, Pteromalidae, Torymidae, Trichogrammatidae, and Bethylidae) and one family of Diptera (Tachinidae). This paper deals with EGVM parasitoids of the families Tachinidae (Diptera) and Braconidae (Hymenoptera). Only two species of Tachinidae are associated to EGVM larvae in Italy, Actia pilipennis (Fallen) and Phytomyptera nigrina (Meigen), whereas the record of Eurysthaea scutellaris (Robineau-Desvoidy) is doubtful. Moreover, 21 species of Braconidae are reported to live on EGVM, but, unfortunately, eight of them were identified only at generic level. Bracon mellitor Say has been incorrectly listed among the parasitoids of Lobesia botrana. Records concerning Ascogaster rufidens Wesmael, Meteorus sp., Microgaster rufipes Nees, and Microplitis tuberculifer (Wesmael) are uncertain.

Catálogo do material-tipo de Tachinidae (Diptera) depositado no Museu de Zoologia da Universidade de São Paulo / Catalogue of the type-material of Tachinidae deposited at the Museu de Zoologia da Universidade de São Paulo

Catálogo do material-tipo de Tachinidae (Diptera) depositado no Museu de Zoologia da Universidade de São Paulo. O Museu de Zoologia da Universidade de São Paulo abriga a maior e mais representativa coleção brasileira de Diptera. A coleção de Tachinidae destaca-se como a segunda mais numerosa, com 32554 espécimes adultos montados. No presente estudo é apresentado um catálogo de tipos primários e secundários de Tachinidae abrigados no Museu de Zoologia, fornecendo informações sobre dados de coleta, seu estado de conservação, bem como o status taxonômico dos nomes específicos e seu atual posicionamento genérico, quando diferente do original. O catálogo lista um total de 847 espécimes-tipo (99 holótipos, 737 parátipos, 8 síntipos, 1 lectótipo, 1 paralectótipo e 1 neótipo) representando um total de 263 espécies nominais de Tachinidae, predominantemente da Região Neotropical. Também são listados espécimes cujo status (se tipo ou não) foi considerado duvidoso. Uma breve biografia de Charles Henry Tyler Townsend e José Henrique Guimarães é apresentada.

Catalogue of the type-material of Tachinidae deposited at the Museu de Zoologia da Universidade de São Paulo. The Museu de Zoologia da Universidade de São Paulo (Brazil) houses the largest and most representative Brazilian collection of Diptera. The Tachinidae collection appears as the second most numerous, with a total of 32554 mounted adult specimens. In the present study, it is presented a catalogue of primary and secondary type-specimens deposited at the Museu de Zoologia, providing information on collecting data, preserving conditions, as well as the taxonomic status of specific names and its current generic placement, whenever different from the original combination. The catalogue lists a total of 847 type-specimens (99 holotypes, 737 paratypes, 8 syntypes, 1 lectotype, 1 paralectotype and 1 neotype) representing 263 nominal species of Tachinidae, mostly from the Neotropical region. Also, the specimens with dubious status (if type or not) are listed. A short biography of Charles Henry Tyler Townsend and José Henrique Guimarães is presented.