First record of Buninotus Maldonado Capriles and B.palikur Castro-Huertas, Forero & Melo from Brazil, with taxonomic notes (Hemiptera, Reduviidae, Emesinae) and an updated key to the genera of Saicini of the New World.

Buninotus Maldonado Capriles, 1981 and Buninotuspalikur Castro-Huertas, Forero & Melo, 2022 (Hemiptera, Reduviidae, Emesinae, Saicini) are recorded from Brazil for the first time. Taxonomic notes on Buninotus and its species are provided mainly based on the examination and photographs of the holotype and paratype of Buninotusniger Maldonado Capriles, 1981. Previous doubts on some characteristics of the genus are clarified. A hypothesis suggesting that the holotype and paratype of B.niger may belong to different species is presented. An updated key to the New World genera of Saicini is provided.

Past, present and future of the two-spotted stink bug (Perillus bioculatus) in Europe revealed by citizen science.

The introduction of the Nearctic predaceous stink bug species, (Perillus bioculatus) was attempted multiple times in various countries throughout Europe to mitigate the damage caused by the invasive and harmful pest species, the Colorado potato beetle (Leptinotarsa decemlineata). Though these attempts were thought to be unsuccessful for decades, more recent data elucidated that the species have established small self-sustaining populations in the Balkans Peninsula, Southern Russia, and Türkiye and recently began to expand. In the past years, the European range of the species reached Eastern Europe. After the first individuals were found in Hungary in October 2023 a citizen science campaign was launched to investigate the distribution of the species in the country. By June 2024 it became evident that the species is established throughout the country. Furthermore, observations regarding beetle larvae and moth caterpillars as alternative prey were reported supporting the previous assumptions that the naturalization and expansion of the species in Europe is facilitated by dietary drift. Here, we summarize the knowledge on the European presence of the two-spotted stink bug and formulate hypotheses regarding its future distribution and the impact of the species on the insect communities of the newly colonized areas.

The redescription and complete mitogenomes of two Oxycarenus species (Hemiptera, Oxycarenidae) and phylogenetic implications.

In this study, the two Oxycarenidae species, O.gossypii Horváth, 1926 and Oxycarenusbicolorheraldus Distant, 1904, are redescribed, and their complete mitogenomes are sequenced and analyzed. The phylogeny of Lygaeoidea is examined using 45 complete mitogenomes of lygaeoid species and four outgroup species. The gene orientation and arrangement of the two mitogenomes are found to be consistent with typical Lygaeoidea mitochondrial features, comprising 37 genes, including 13 PCGs, 22 tRNAs, 2 rRNAs, and a control region. Nucleotide composition of the species was biased towards A and T, with the gene order identical to the putative ancestral arrangement of insects. Start codons, stop codons, RNAs, relative synonymous codon usage (RSCU), and nucleotide diversity (Pi) of Oxycarenidae exhibited characteristics similar to other families in Lygaeoidea. Bayesian-inference (BI) and maximum-likelihood (ML) methods were employed to investigate phylogenetic relationships using PCG datasets from selected species. Phylogenetic analyses reveal slightly different topologies between BI and ML methods, with variation primarily concentrated in Colobathristidae and Rhyparochromidae. Our study confirms that the two sequenced Oxycarenidae species formed a single clade, and the position of Oxycarenidae remains stable in both ML and BI phylogenetic trees. These findings expand the mitochondrial genome databases of Lygaeoidea and provide valuable insights into the phylogenetic relationships within Lygaeoidea or Pentatomomorpha.

The First FISH-Confirmed Non-Canonical Telomeric Motif in Heteroptera: Cimex lectularius Linnaeus, 1758 and C. hemipterus (Fabricius, 1803) (Hemiptera, Cimicidae) Have a 10 bp Motif (TTAGGGATGG)n.

Fluorescence in situ hybridization (FISH) with two different probes, the canonical insect telomeric sequence (TTAGG)n and the sequence (TTAGGGATGG)n, was performed on meiotic chromosomes of two members of the true bug family Cimicidae (Cimicomorpha), the common bed bug Cimex lectularius Linnaeus, 1758 and the tropical bed bug C. hemipterus (Fabricius, 1803), whose telomeric motifs were not known. In both species, there were no hybridization signals with the first probe, but strong signals at chromosomal ends were observed with the second probe, indicating the presence of a telomeric motif (TTAGGGATGG)n. This study represents the first FISH confirmation of the presence of a non-canonical telomeric motif not only for the infraorder Cimicomorpha but also for the suborder Heteroptera (Hemiptera) as a whole. The present finding is of key significance for unraveling the evolutionary shifts in the telomeric sequences in this suborder.

The burrower bug Macroscytus japonensis (Hemiptera: Cydnidae) acquires obligate symbiotic bacteria from the environment.

Many plant-feeding stinkbugs belonging to the infraorder Pentatomomorpha possess a specialized symbiotic organ at the posterior end of the midgut, in which mutualistic bacterial symbionts are harbored extracellularly. In species of the superfamily Pentatomoidea, these symbionts typically are verticallytransmitted from host mothers to offspring, whereas in species of the superfamilies Coreoidea and Lygaeoidea they are acquired from the environment. In the pentatomoid family Cydnidae, vertical symbiont transmission has been reported in several species. Here, we report the first case of environmental symbiont acquisition in Cydnidae, observed in the burrower bug Macroscytus japonensis. A comprehensive survey of 72 insect samples from 23 sites across the Japanese archipelago revealed that (1) symbionts exhibit remarkably high diversity, forming six distinct phylogenetic groups within the Enterobacteriaceae of the γ-Proteobacteria, (2) most symbionts are cultivable and closely related to free-living Pantoea-allied bacteria, and (3) symbiont phylogenetic groups do not reflect the host phylogeny. Microbial inspection of eggs revealed the absence of bacteria on the egg surface. These results strongly suggest that symbionts are acquired from the environment, not vertical transmission. Rearing experiments confirmed environmental symbiont acquisition. When environmental symbiont sources were experimentally withheld, nymphs became aposymbiotic and died before molting to the second instar, indicating that nymphs environmentally acquire symbionts during the first-instar stage and that symbionts are essential for nymphal growth and survival. This study highlights Cydnidae as the only pentatomoid family that includes species that environmentally acquire symbionts and those that vertically transmit symbionts, providing an ideal platform for comparative studies of the ecological and environmental factors that influence the evolution of symbiont transmission modes.

A new species of Neotropiconyttus Kirkaldy (Hemiptera, Reduviidae) in a cacao plantation from the Colombian Napo Province, including a key to species, and taxonomic notes of the genus.

A new species of the genus Neotropiconyttus Kirkaldy, 1909 (Hemiptera: Heteroptera: Reduviidae: Harpactorinae: Harpactorini) is described and illustrated. Neotropiconyttusarmandoi Gamboa & Gil-Santana, sp. nov. represents the first record of the genus for the Province of Napo in Colombia, and the first description of a male individual in the genus. The male specimen representing the new species was collected on a leaf of cacao (Theobromacacao L.-Malvaceae). Its remarkable similarity in external coloration and structure with that of the true bug Monaloniondissimulatum Distant, 1883 (Hemiptera: Miridae) inhabiting cacao agroforestry systems suggests that the new species could be part of a mimetic complex that incorporates phytophagous and predator bugs. Comments and figures of type specimens of Neotropiconyttusalboannulatus (Stål, 1855) and Neotropiconyttusdama (Burmeister, 1838), and a key to the species of the genus are also provided.

Structure and bioactivity of an insecticidal trans-defensin from assassin bug venom.

Disulfide-rich peptides such as defensins play diverse roles in immunity and ion channel modulation, as well as constituting the bioactive components of many animal venoms. We investigated the structure and bioactivity of U-RDTX-Pp19, a peptide previously discovered in venom of the assassin bug Pristhesancus plagipennis. Recombinant Pp19 (rPp19) was found to possess insecticidal activity when injected into Drosophila melanogaster. A bioinformatic search revealed that domains homologous to Pp19 are produced by assassin bugs and diverse other arthropods. rPp19 co-eluted with native Pp19 isolated from P. plagipennis, which we found is more abundant in hemolymph than venom. We solved the three-dimensional structure of rPp19 using 2D 1H NMR spectroscopy, finding that it adopts a disulfide-stabilized structure highly similar to known trans-defensins, with the same cystine connectivity as human α-defensin (I-VI, II-IV, and III-V). The structure of Pp19 is unique among reported structures of arthropod peptides.

Halyomorpha halys (Hemiptera: Pentatomidae) as the major contributor to early olive drop in northern Italy.

In recent years, a new phenomenon of early olive drop is causing production losses in olive groves throughout northern Italy. To analyze the possible causes, field and laboratory trials were performed to assess the involvement of fungal pathogens and insect pests in this disease. External and internal symptoms of fungal infections or insect-feeding activities were researched. Fungi present in healthy and dislodged olives were investigated. The relationship between olives that fell and Halyomorpha halys (Stål) (Hemiptera: Pentatomidae) infestation was assessed in a controlled infestation trial, and the effectiveness of an insecticidal strategy in reducing early olive drop was tested in open field conditions. A comparable number of fungi, mostly endophytes, were isolated and identified from both healthy and dislodged olives. The damage observed on dislodged olives was primarily ascribed to pentatomids feeding activity. Six stink bugs species were found in olive canopies, that is, the invasive H. halys, which was by far the most abundant, and Acrosternum heegeri Fieber, Nezara viridula (Linnaeus), Palomena prasina (Linnaeus), Piezodorus lituratus (Fabricious), and Rhaphigaster nebulosa (Poda). Halyomorpha halys caused intense fruit drop in the controlled infestation trial, and its infestation level significantly correlated with the number of olives that fell. Native stink bugs, present in much lower population compared to H. halys, could also partially contribute to early drop of olives. Insect proof net significantly reduced the early olive drop disease, while insecticide applications only partially reduced the stink bugs population density and, proportionally, early olive drop.

Multiple and frequent trypanosomatid co-infections of insects: the Cuban case study.

Trypanosomatids are obligate parasites of animals, predominantly insects and vertebrates, and flowering plants. Monoxenous species, representing the vast majority of trypanosomatid diversity, develop in a single host, whereas dixenous species cycle between two hosts, of which primarily insect serves as a vector. To explore in-depth the diversity of insect trypanosomatids including their co-infections, sequence profiling of their 18S rRNA gene was used for true bugs (Hemiptera; 18% infection rate) and flies (Diptera; 10%) in Cuba. Out of 48 species (molecular operational taxonomic units) belonging to the genera Vickermania (16 spp.), Blastocrithidia (7), Obscuromonas (4), Phytomonas (5), Leptomonas/Crithidia (5), Herpetomonas (5), Wallacemonas (2), Kentomonas (1), Angomonas (1) and two unnamed genera (1 + 1), 38 species have been encountered for the first time. The detected Wallacemonas and Angomonas species constitute the most basal lineages of their respective genera, while Vickermania emerged as the most diverse group. The finding of Leptomonas seymouri, which is known to rarely infect humans, confirms that Dysdercus bugs are its natural hosts. A clear association of Phytomonas with the heteropteran family Pentatomidae hints at its narrow host association with the insect rather than plant hosts. With a focus on multiple infections of a single fly host, using deep Nanopore sequencing of 18S rRNA, we have identified co-infections with up to 8 trypanosomatid species. The fly midgut was usually occupied by several Vickermania species, while Herpetomonas and/or Kentomonas species prevailed in the hindgut. Metabarcoding was instrumental for analysing extensive co-infections and also allowed the identification of trypanosomatid lineages and genera.

New species of Hexacladia Ashmead (Hymenoptera: Encyrtidae), a parasitoid of Tibraca limbativentris Stål (Hemiptera: Pentatomidae)

ABSTRACT The rice stink bug, Tibraca limbativentris Stål (Hemiptera: Pentatomidae), is a significant pest of rice cultivation in Brazil and attacks plants during both vegetative and reproductive phases. Natural enemy surveys conducted in plantations located in Arari, Miranda do Norte, Matões do Norte, Santa Rita, and Viana (state of Maranhão, Brazil) led to the discovery of a new species of parasitoid, which is described in this study. Hexacladia lemosae Costa, Noyes & Machado sp. nov. (Hymenoptera: Encyrtidae) is the first species of its genus associated with T. limbativentris and develops as a gregarious endoparasitoid in adult stink bugs, occasionally in nymphs. The parasitoid emerges from the host while host is still alive.

Revisiting the Taxonomy of Cylapocoris Carvalho, 1954 (Hemiptera: Miridae: Cylapinae) with Descriptions of Five New Species and Morphology-Based Phylogenetic Analysis of the Genus.

This paper provides descriptions of five new species of the Neotropical genus Cylapocoris Carvalho, 1954 (C. bimaculatus n. sp., C. brooksi n. sp., C. carvalhoi n. sp., C. scutellatus n. sp., and C. simplexoides n. sp.). Cylapocoris and Cylapocoroides Carvalho, 1989 are redescribed and rediagnosed. Illustrations of male genitalia, scanning electron micrographs of selected structures of certain taxa, and an identification key to species are provided. Female genitalia are described and illustrated for the first time for Cylapocoris in nine out of 19 known species. A cladistic analysis of the genus, based on 62 morphological characters, is presented as a contribution to the understanding of relationships within Cylapocoris and its relationships with other groups of Cylapinae. The analysis comprises 16 ingroup species and 15 outgroup taxa. Both equal and implied weighting parsimony analyses were used in the phylogenetic reconstruction. We confirm the monophyly of Cylapocoris and its sister-group relationship with Cylapocoroides. Additionally, we identify subgroupings within Cylapocoris. Intertribal relationships within Cylapinae are briefly discussed.

Food habits of 3 myrmecophilous bug species on myrmecophytic Macaranga (Malpighiales: Euphorbiaceae) vary from herbivory to predation.

Myrmecophytes have mutualistic relationships with symbiotic ants. Although myrmecophytic Macaranga (Malpighiales: Euphorbiaceae) species are well protected by aggressive Crematogaster (Hymenoptera: Formicidae) ants, some bug species occur on the myrmecophytes. To clarify the associations of these bugs with the plants and the ants, we studied the food habits of 3 bug species, Pilophorus lambirensis Nakatani et Komatsu, 2013 (Hemiptera: Miridae: Phylinae), Phylinae sp. 1, and Arbela sp. 1 (Hemiptera: Nabidae). We conducted field observations in a Bornean rainforest. First, we located these bugs and studied their behavioral responses to the ants on Macaranga species; we then conducted stable isotope analyses. All bugs avoided direct contact with ants, but they occurred only on trees with active ants. Pilophorus lambirensis and Phylinae sp. 1 were most commonly observed on the apical parts of host trees, whereas Arbela sp. 1 was mainly in areas distant from the apical parts where ants were sparse. The stable isotope ratios indicated that Phylinae sp. 1 fed on food bodies, which are nutrient-rich spherical bodies produced by Macaranga trees on the apical parts for ants. Although the main diet of the other 2 species remains unclear, nitrogen isotopic signatures demonstrated that P. lambirensis is herbivorous, whereas Arbela sp. 1 is carnivorous. However, the distant location from ants and its isotopic signatures indicated that Arbela sp. 1 rarely fed on the ants. At least 2 mirid bug species might obtain enemy-free space in addition to the food provided by the myrmecophytes.

The spread of satellite DNAs in euchromatin and insights into the multiple sex chromosome evolution in Hemiptera revealed by repeatome analysis of the bug Oxycarenus hyalinipennis.

Satellite DNAs (satDNAs) are highly repeated tandem sequences primarily located in heterochromatin, although their occurrence in euchromatin has been reported. Here, our aim was to advance the understanding of satDNA and multiple sex chromosome evolution in heteropterans. We combined cytogenetic and genomic approaches to study, for the first time, the satDNA composition of the genome in an Oxycarenidae bug, Oxycarenus hyalinipennis. The species exhibits a male karyotype of 2n = 19 (14A + 2 m + X1 X2 Y), with a highly differentiated Y chromosome, as demonstrated by C-banding and comparative genomic hybridization, revealing an enrichment of repeats from the male genome. Additionally, comparative analysis between males and females revealed that the 26 identified satDNA families are significantly biased towards male genome, accumulating in discrete regions in the Y chromosome. Exceptionally, the OhyaSat04-125 family was found to be distributed virtually throughout the entire extension of the Y chromosome. This suggests an important role of satDNA in Y chromosome differentiation, in comparison of other repeats, which collectively shows similar abundance between sexes, about 50%. Furthermore, chromosomal mapping of all satDNA families revealed an unexpected high spread in euchromatic regions, covering the entire extension, irrespective of their abundance. Only discrete regions of heterochromatin on the Y chromosome and of the m-chromosomes (peculiar chromosomes commonly observed in heteropterans) were enriched with satDNAs. The putative causes of the intense enrichment of satDNAs in euchromatin are discussed, including the possible existence of burst cycles similar to transposable elements and as a result of holocentricity. These data challenge the classical notion that euchromatin is not enriched with satDNAs.

A Faboideae-Specific Floral Scent Betrays Seeds to an Important Granivore Pest.

Seed predation by insect herbivores reduces crop production worldwide. Foraging on seeds at pre-dispersal generally means that females need to find the suitable host plant within a relatively short timeframe in order to synchronize larval development with seed production. The mechanistic understanding of host finding by seed pests can be harnessed for more sustainable pest management strategies. We here studied the chemical communication between the bean bug Riptortus pedestris, a major pest of legumes, and several crop species and cultivars in the Fabaceae. Via a comparative chemical analysis, we found that 1-octen-3-ol is the principal constituent of the floral scents of most species tested in the subfamily Faboideae, including soybean and faba bean. With field trapping and laboratory bioassays, including electroantennography, we further revealed that this compound can be perceived, and stimulate attraction responses, by R. pedestris nymphs and adults. The addition of 1-octen-3-ol to pheromone traps might therefore improve trapping efficacy for controlling populations of this important granivore pest on legumes.

A Revision of the Genus Argolis (Hemiptera: Reduviidae: Stenopodainae) from Asia.

The assassin bug genus Argolis Stål, 1861 (Hemiptera: Reduviidae: Stenopodainae) has a disjunct distribution in Sub-Saharan Africa and Asia. In the present study, the Asian species of Argolis are revised. Two species are recognized, redescribed, and illustrated, with the following new subjective synonyms and new combination proposed: Argolis Stål, 1861 = Bardesanes Distant, 1909, syn. nov. = Neoklugia Distant, 1919, syn. nov.; A. farinator (Reuter, 1882) = N. typica Distant, 1919, syn. nov. = B. sericenotatus Livingstone & Ravichandran, 1989, syn. nov.; A. signata (Distant, 1909), comb. nov. (transferred from Bardesanes) = Caunus noctulus Hsiao, 1977, syn. nov. Lectotypes for C. farinator, B. signatus, and N. typica are designated. A key to separate the two Asian species of Argolis is provided. The sexual dimorphism, systematic relationships, and distribution of Argolis are discussed. Argolis is newly recorded from Laos, Pakistan, and Vietnam.

Ontogenetic change in effectiveness of chemical defence against different predators in Oxycarenus true bugs.

Many prey species change their antipredator defence during ontogeny, which may be connected to different potential predators over the life cycle of the prey. To test this hypothesis, we compared reactions of two predator taxa - spiders and birds - to larvae and adults of two invasive true bug species, Oxycarenus hyalinipennis and Oxycarenus lavaterae (Heteroptera: Oxycarenidae) with life-stage-specific chemical defence mechanisms. The reactions to larvae and adults of both true bug species strikingly differed between the two predator taxa. The spiders were deterred by the defences of adult bugs, but the larval defences were ineffective against them. By contrast, birds attacked the larvae considerably less often than the adult bugs. The results indicate a predator-specific ontogenetic change in defence effectiveness of both Oxycarenus species. The change in defence is likely linked to the life-stage-specific composition of secretions in both species: whereas secretions of larvae are dominated by unsaturated aldehydes, secretions of adults are rich in terpenoids, which probably serve dual function of defensive chemicals and pheromones. Our results highlight the variation in defence between different life stages and the importance of testing responses of different types of predators.

Characterization of two new Pylorgus mitogenomes (Hemiptera, Lygaeidae, Ischnorhynchinae) and a mitochondrial phylogeny of Lygaeoidea.

Lygaeidae is a large family of Hemiptera (Heteroptera) currently separated into three subfamilies, Ischnorhynchinae, Lygaeinae, and Orsillinae. In this research, the complete mitogenomes of the iscnorhynchines Pylorgusporrectus Zheng, Zou & Hsiao, 1979 and Pylorgussordidus Zheng, Zou & Hsiao, 1979 were sequenced, and the phylogeny of Pylorgus and the Lygaeidae with known complete mitogenomes were examined. The mitogenomes are 15,174 bp and 15,399 bp in size, respectively, and comprised of 13 protein-coding genes (PCGs), 22 transfer RNA genes (tRNAs), two ribosomal RNA genes (rRNAs), and a control region (D-loop). Nucleotide composition is biased toward A and T, and the gene order is identical to that of the putative ancestral arrangement of insects. Eleven PCGs begin with a typical ATN, and the remaining two PCGs begin with TTG (cox1 and nad4l). All tRNAs had a typical cloverleaf secondary structure, but some of them had individual base mismatches. The phylogenetic analyses based on the concatenated nucleotide sequences of the 13 PCGs, using Bayesian inference and maximum likelihood, support the monophyly of Lygaeidae. The results show that P.porrectus and P.sordidus clustered with nine other Lygaeidae. This study includes the first complete sequencing of the mitochondrial genomes of two Pylorgus species, which will provide important data for studying the phylogenetic position of Lygaeidae in Lygaeoidea and reconstructing the phylogenetic relationships within Pentatomomorpha.

Reproductive biology of overwintering leaffooted bug Leptoglossus zonatus (Hemiptera: Coreidae) in California.

Leptoglossus zonatus (Dallas) (Hemiptera: Coreidae) is a polyphagous insect pest attacking a wide variety of crops. In California's Central Valley, it is now the dominant leaffooted bug on almonds, pistachios, and pomegranates. Leptoglossus zonatus pest status depends largely on overwintering adult survival and reproductive potential, which determines its population size in spring and early summer when nut crops are particularly susceptible to bug damage. Here, we investigated the overwintering reproductive biology of L. zonatus in laboratory and field experiments to gain information about its ovary development, time of mating, and the impact of low temperatures on egg hatch. With dissections of laboratory-reared L. zonatus, we established a baseline for ovarian development and determined that the size of the spermathecal reservoir is larger in mated than in unmated females. Dissections and behavioral experiments of field-collected material provided evidence of mating events before dispersal from overwintering sites. Laboratory trials showed that temperature significantly impacted L. zonatus egg hatch. Leptoglossus zonatus reproductive biology presented provides valuable information on its population dynamics and dispersal from overwintering sites, and will contribute to the development of monitoring and management tools.

First record of the predatory stink bug species Picromerusgriseus (Dallas) (Hemiptera, Heteroptera, Pentatomidae, Asopinae) in Japan, with an illustrated key to the Japanese species of the genus Picromerus Amyot & Serville.

Background: The predatory stink bug genus Picromerus Amyot & Serville, 1843 (Hemiptera, Heteroptera, Pentatomidae, Asopinae) comprises 11 species found in the Northern Hemisphere. In Japan, two species have been recorded to date. However, an easy-to-understand identification method, such as an illustrated key, is lacking. Currently, Picromerusgriseus (Dallas, 1851) has been recorded in Bangladesh, Bhutan, China, Indonesia, Myanmar, Pakistan and Taiwan, but not in Japan. New information: Picromerusgriseus was recorded in Japan for the first time, based on a single individual collected from grasslands around the fields of Ishigaki Island of the Ryukyu Islands, which belong to the Oriental Region. This discovery represents the easternmost record of the species. An illustrated key to the species of Picromerus occurring in Japan is also provided.

Juvenile hormone identification in the cabbage bug Eurydema rugosa.

Juvenile hormone (JH) plays a pivotal role in almost every aspect of insect development and reproduction. The chemical structure of the JH in heteropteran species has long remained elusive until methyl (2R,3S,10R)-2,3;10,11-bisepoxyfarnesoate, commonly named as juvenile hormone III skipped bisepoxide (JHSB3), was isolated from Plautia stali (Hemiptera: Heteroptera: Pentatomidae). Recently, several groups reported the presence of JHSB3 in other heteropteran species. However, most of the studies paid no attention to the determination of the relative and absolute structure of the JH. In this study, we investigated the JH of the cabbage bug Eurydema rugosa (Hemiptera: Heteroptera: Pentatomidae), known as a pest for wild and cultivated crucifers. JHSB3 was detected in the hexane extract from the corpus allatum (CA) product using a chiral ultraperformance liquid chromatography-tandem mass spectrometer (UPLC-MS/MS) which can inform the absolute stereochemistry of the JH. Its stereoisomers were not detected. Topical application of the synthetic JHSB3 to the last instar nymphs inhibited their metamorphosis and induced nymphal-type colouration of the dorsal abdomen in a dose-dependent manner. Additionally, the topical application of JHSB3 effectively terminated summer and winter diapauses in females. These results indicate that the JH of E. rugosa is JHSB3. Although individuals in summer and winter diapauses are physiologically distinct in E. rugosa, the results suggest that the physiological differences between these diapauses are based, not on the responsiveness to JH, but on the processes governing activation of the CA or on its upstream cascades.