Search

The stink bugs Acrosternum arabicum Wagner and Brachynema germari Kolenati (Hemiptera: Pentatomidae) damage developing pistachio nuts in Iran. Ooencyrtus egeria Huang and Noyes (Hymenoptera: Encyrtidae) parasitizes eggs of both species, alternating between them seasonally. We compared the biological and life history attributes of O. egeria reared for 3 generations on each host species with those that had been transferred to the alternate host for 2 generations. We also tested female responses to host egg density. Eggs of B. germari were parasitized at higher rates and yielded faster wasp development, heavier female progeny with greater fecundity, and a more female-biased sex ratio, than eggs of A. arabicum. Rearing on A. arabicum did not diminish wasp fitness on B. germari, and the F1 progeny of wasps switched to A. arabicum inherited some beneficial parental effects from rearing on B. germari. Regardless of host species, females parasitized fewer eggs in 8 h as density increased from 10 to 30 eggs, but progeny sex ratio (% female) increased. The anomalous response to host density appears to arise from a combination of behavioral and egg load constraints. Females mature only 10-12 eggs per day. Carefully inspect egg clusters before selecting the most suitable eggs for oviposition, a more time-consuming process for large clusters. Our results indicate that rearing O. egeria on eggs of A. arabicum, which are more convenient and cost-effective to produce than those of B. germari, will not diminish its performance on B. germari following augmentative release, beyond a slight reduction in female fecundity., (© The Author(s) 2024. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Background: Plant volatile organic compounds (VOCs) play a crucial role in mediating interactions between plants, herbivores and natural enemies. Among these VOCs, methyl salicylate and (E,E)-α-farnesene are emitted as herbivore-induced plant volatiles (HIPVs) by soybean plants in response to feeding by the brown stink bug Eushistus heros. These HIPVs function as synomones, influencing the foraging behaviour of the egg parasitoid, Telenomus podisi, the main natural enemy of E. heros, one of the major soybean pests in Brazil., Results: Laboratory experiments showed that two soybean cultivars, BRS 7580 and BRS 7880, produced similar qualitative blends of HIPVs, with methyl salicylate, (E,E)-α-farnesene and (Z)-3-hexenyl acetate being produced by both cultivars. Soybean cultivar BRS 7580 produced a significant lower amount of HIPVs compared to BRS 7880 but this difference did not affect the attractiveness of the egg parasitoid Telenomus podisi. Field experiments using these two cultivars and synthetic applications of methyl salicylate and (E,E)-α-farnesene showed a substantial increase in egg parasitism in all treated areas. Parasitism rates ranged from 50% to 80% in areas where these HIPVs were deployed, compared to only 10% in untreated control areas., Conclusions: The egg parasitoid Telenomus podisi demonstrated an adept ability in recognising between HIPVs in soybean blends, even in the presence of significant quantitative differences. The results from the field experiment showed the potential of HIPVs in attracting natural enemies to specific target areas within fields. (E,E)-α-Farnesene showed an improved action during the later stages of soybean growth, notably at R6. In addition, this volatile attracted other families of natural enemies. © 2024 Society of Chemical Industry., (© 2024 Society of Chemical Industry.)

Fall armyworm (FAW), Spodoptera frugiperda is a generalist pest known to feed on more than 300 plant species, including major staple crops such as rice, maize and sorghum. Biological control of FAW using a combination of a major indigenous egg parasitoid Telenomus remus and entomopathogenic fungi was explored in this study. Metarhizium anisopliae strains (ICIPE 7, ICIPE 41, and ICIPE 78) and Beauveria bassiana ICIPE 621 which demonstrated effectiveness to combat the pest, were evaluated through direct and indirect fungal infection to assess their pathogenicity and virulence against T. remus adults, S. frugiperda eggs and their effects on T. remus parasitism rates. Metarhizium anisopliae ICIPE 7 and ICIPE 78 exhibited the highest virulence against T. remus adults with LT 50 values >2 days. ICIPE 7 induced the highest T. remus mortality rate (81.40 ± 4.17%) following direct infection with dry conidia. Direct fungal infection also had a significant impact on parasitoid emergence, with the highest emergence rate recorded in the M. anisopliae ICIPE 7 treatment (42.50 ± 5.55%), compared to the control ± (83.25 ± 5.94%). In the indirect infection, the highest concentration of 1 x 10 9 conidia ml -1 of ICIPE 78 induced the highest mortality (100 ± 0.00%) of T. remus adults, and the highest mortality (51.25%) of FAW eggs, whereas the least FAW egg mortality (15.25%) was recorded in the lowest concentration 1 x 10 5 conidia ml -1 of ICIPE 41. The number of parasitoids that emerged and their sex ratios were not affected by the different fungal strain concentrations except in ICIPE 7 at high dose. This study showed that potential combination of both M. anisopliae and B. bassiana with T. remus parasitoid can effectively suppress FAW populations., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2024 Chepkemoi, Fening, Ambele, Munywoki and Akutse.)

The use of biocontrol agents, such as predators and entomopathogenic nematodes, is a promising approach for the effective control of the tomato leafminer Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidaean), an oligophagous insect feeding mainly on Solanaceae species and a major pest of field- and greenhouse-grown tomatoes globally. In this context, the effects of two entomopathogenic nematode species Steinernema carpocapsae (Weiser) (Rhabditida: Steinernematidae) and Heterorhabditis bacteriophora (Poinar) (Rhabditida: Heterorhabditidae), as well as their respective bacterial symbionts, Xenorhabdus nematophila and Photorhabdus luminescens (Enterobacterales: Morganelaceae), which were applied as bacterial cell suspensions and as crude cell-free liquid filtrates on T. absoluta larvae, were investigated. The results showed that of all treatments, the nematodes S. carpocapsae and H. bacteriophora were the most effective, causing up to 98 % mortality of T. absoluta larvae. Regarding bacteria and their filtrates, the bacterium X. nematophila was the most effective (69 % mortality in young larvae), while P. luminescens and both bacterial filtrates showed similar potency (ca. 48-55 % mortality in young larvae). To achieve a holistic approach of controlling this important pest, the impact of these factors on the beneficial predator Nesidiocoris tenuis (Reuter) (Hemiptera: Miridae) was also studied. The results demonstrated that although nematodes and especially S. carpocapsae, caused significant mortality on N. tenuis (87 %), the bacterial cell suspensions of X. nematophila and P. luminescens and crude cell-free liquid filtrates had minimum impact on this beneficial predator (∼11-30 % mortality)., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 The Author(s). Published by Elsevier Inc. All rights reserved.)

Parasites are known to have direct effects on host dispersal ability and motivation. Yet, parasites have a variety of impacts on host populations, including shaping predation and cannibalism rates, and therefore may also have indirect effects on host dispersal; these indirect pathways have not been studied. We tested the hypothesis that parasites influence host dispersal through effects on cannibalism using backswimmers ( Notonecta undulata ) and Hydrachnidia freshwater mites. Mite parasitism impedes swimming in backswimmers, which we found increased their vulnerability to cannibalism. We imposed a manipulation that varied cannibalism rates across experimental populations consisting of a mix of backswimmers with and without simulated parasites. Using simulated parasites allowed us to examine the effects of cannibalism without introducing infection risk. We found that the odds of dispersal for infected backswimmers increased by 2.25× with every 10% increase in the risk of being cannibalized, and the odds of dispersal for healthy backswimmers increased by 2.34× for every additional infected backswimmer they consumed. Our results suggest that cannibalism was used as an energy source for dispersal for healthy individuals, while the risk of being eaten motivated dispersal in infected individuals. These results elucidate the complex ways that parasites impact host populations and strengthen our understanding of host-parasite interactions, including host and parasite population stability and spread. This article is part of the theme issue 'Diversity-dependence of dispersal: interspecific interactions determine spatial dynamics'.

Wasps are important parasitoids of stinkbugs and frequently exposed to various types of microorganisms through environmental contact and fecal-oral transmission route. Many parasitize stinkbug eggs and are commercially used in the field to control insect population. The parasitoid T. podisi is known for its high parasitism capacity and ability to target multiple species of stinkbugs. In this study we asked whether T. podisi exposed to eggs infected by a multispecies asymptomatic stinkbug virus, the Halyomorpha halys virus (HhV) would get infected. HhV is a geographically distributed multispecies iflavirus previously found to infect four stinkbug hosts, including three Brazilian species, Chinavia ubica, Euschistus heros and Diceraeus melacanthus, and T. posidi can parasitize all of them. As results, RT-PCR screening revealed positive samples for the HhV genome in two out of four tested pools of T. podisi, whereas the antigenome, indicative of replicative activity, was not detected. The wasps were raised in E. heros eggs that presented both the genome and the antigenome forms of the HhV genome. Subsequent RNA-deep sequencing of HhV positive T. podisi RNA pools yielded a complete genome of HhV with high coverage. Phylogenetic analysis positioned the isolate HhV-Tp (isolate Telenomus podisi) alongside with the stinkbug HhV. Analysis of transcriptomes from several hymenopteran species revealed HhV-Tp reads in four species. However, the transmission mechanism and the ecological significance of HhV remain elusive, warranting further studies to illuminate both the transmission process and its capacity for environmental propagation using T. podisi as a potential vector., (© 2024. The Author(s) under exclusive licence to Sociedade Brasileira de Microbiologia.)

In this work, we investigated parasites of the firebug Pyrrhocoris apterus in Austria and demonstrated that in addition to the extensively studied Leptomonas pyrrhocoris, it can also be infected by Blastocrithidia sp. and by a mermithid, which for the first time has been characterized using molecular methods. This diversity can be explained by the gregarious lifestyle, as well as the coprophagous and cannibalistic behavior of the insect hosts that makes them susceptible to various parasites. In addition, we showed no tight association of the L. pyrrhocoris haplotypes and geographical locations (at least, considering the relatively small scale of locations in Austria) implying that the natural populations of L. pyrrhocoris are mixed due to the mobility of their firebug hosts., (© 2023. The Author(s).)

Non-cultivated areas are resting, overwintering, feeding, and/or reproducing habitats for insects, and also places from where crop areas are colonized; thus, they are essential for understanding the biological control programs in agroecosystems. We developed a simulation model for a non-cultivated area of Buenos Aires province (Argentina), and we analyzed the control of Nezara viridula achieved by the action of two parasitoids: the oophagous Trissolcus basalis and the tachinid Trichopoda giacomellii , which attack older nymphs and adults. The model is a discrete time, deterministic, phenomenological, spatially homogeneous with a 1-week time interval simulation model, based on the age-structure and/or stage-structure of N. viridula and its two parasitoids. The host-parasitoid interactions were combined with a degree-day model affecting development times of T. giacomellii pupae and T. basalis pre-imaginal stages. The simultaneous attack of both parasitoid species enables the persistence of the system at low host densities, mediated by the functional response of the parasitoids, identified as population regulation factors. However, if only one parasitoid exists (i.e., only T. basalis or only T. giacomellii ) the interaction N. viridula -parasitoid persisted but at higher density of N. viridula . These results explain the successful biological control of N. viridula after the introduction of T. basalis in the 1980s, when T. giacomellii was the only parasitoid present, unable to control N. viridula . Our model shows an indirect competition when both parasitoids are present: the attack of one of them diminished the potential number of hosts available to the other parasitoid species. In the field this interaction is obscured by the hibernation period which acted as a reset mechanism affecting the density and age/stage structure of all three populations. Our model was supported by field observations, and never exhibited the extinction of any of the parasitoids from the interaction.

Background: Interspecific competition in insect parasitoids is an important ecological phenomenon that has relevant implications for biological pest control. To date, interspecific intrinsic (=larval) competition has been intensively studied, while investigations on extrinsic (=adult) competition have often lagged behind. In this study we examined the role played by parasitoid reproductive traits and host clutch size on the outcome of extrinsic competition between Trissolcus basalis (Wollaston) and Ooencyrtus telenomicida (Vassiliev), two egg parasitoids of the pest Nezara viridula (L). Laboratory experiments were conducted by allowing both parasitoid species to exploit an egg mass made of 10, 20, 30, or 40 hosts through single or simultaneous releases. Furthermore, under field conditions, egg masses consisting of 10 or 40 hosts were exposed in a tomato crop in order to validate laboratory investigation., Results: The results show that the egg mass size is an important predictor of extrinsic competition in our study system as a higher proportion of T. basalis emerged from large egg masses, while O. telenomicida dominated in small egg masses. Analysis of reproductive traits of parasitoid species indicates that T. basalis has superior abilities in host exploitation compared with O. telenomicida., Conclusions: We found that contrasting reproductive traits of two competing egg parasitoid species facilitate coexistence on a shared stink bug host. This work also highlights the importance to consider extrinsic competitive interactions between parasitoid species in a biological control perspective. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry., (© 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.)

Phytomonas serpens is a protozoan parasite that alternates its life cycle between two hosts: an invertebrate vector and the tomato fruit. This phytoflagellate is able to synthesize proteins displaying similarity to the cysteine peptidase named cruzipain, an important virulence factor from Trypanosoma cruzi , the etiologic agent of Chagas disease. Herein, the growth of P. serpens in complex medium (BHI) supplemented with natural tomato extract (NTE) resulted in the increased expression of cysteine peptidases, as verified by the hydrolysis of the fluorogenic substrate Z-Phe-Arg-AMC and by gelatin-SDS-PAGE. Phytoflagellates showed no changes in morphology, morphometry and viability, but the proliferation was slightly reduced when cultivated in the presence of NTE. The enhanced proteolytic activity was accompanied by a significant increase in the expression of cruzipain-like molecules, as verified by flow cytometry using anti-cruzipain antibodies. In parallel, parasites incubated under chemically defined conditions (PBS supplemented with glucose) and added of different concentration of NTE revealed an augmentation in the production of cruzipain-like molecules in a typically dose-dependent way. Similarly, P. serpens recovered from the infection of mature tomatoes showed an increase in the expression of molecules homologous to cruzipain; however, cells showed a smaller size compared to parasites grown in BHI medium. Furthermore, phytoflagellates incubated with dissected salivary glands from Oncopeltus fasciatus or recovered from the hemolymph of infected insects also showed a strong enhance in the expression of cruzipain-like molecules that is more relevant in the hemolymph. Collectively, our results showed that cysteine peptidases displaying similarities to cruzipain are more expressed during the life cycle of the phytoflagellate P. serpens both in the invertebrate and plant hosts., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2022 Oliveira, Elias, Dias, Lopes, d’Avila-Levy, Santos and Branquinha.)

The invasive Halyomorpha halys (Stål) is a significant agricultural and urban nuisance pest in many parts of the world. In North America, biological control of H. halys by parasitoid wasps in the families Scelionidae and Eupelmidae has shown promise. An effective technique for detection and monitoring native and exotic parasitoids is the deployment of yellow sticky cards; however, yellow cards also attract nontarget arthropods, reducing efficiency and accuracy of parasitoid screening. This study sought to identify an alternative yet effective trapping technique by comparing the number of target parasitoid wasps [Anastatus spp. Motschulsky (Hymenoptera: Eupelmidae), Telenomus spp. Haliday (Hymenoptera: Scelionidae), and Trissolcus spp. Ashmead (Hymenoptera: Scelionidae)] and arthropod bycatch on yellow and blue sticky cards deployed in urban, orchard, and vegetable landscapes in northern Utah from late May to early October in 2019 and 2020. Yellow sticky cards captured 54-72% more target parasitoids than blue cards from June through August in all three landscape types in both years; however, a positive correlation in parasitoid capture indicated blue cards detect target parasitoids, just in fewer numbers. Both card colors detected adventive Trissolcus japonicus (Ashmead) (Hymenoptera: Scelionidae) in initial findings of 2019, and in expanded locations of 2020. Furthermore, blue cards captured 31-48% less Diptera and nontarget Hymenoptera than yellow cards in both years across all three landscapes, translating to reduced card processing time and impacts to beneficial insect populations. Our results suggest that blue vs yellow sticky cards offer an alternative monitoring technique to survey for H. halys parasitoids., (© The Author(s) 2021. Published by Oxford University Press on behalf of Entomological Society of America.)

The expanding phylogenetic tree of trypanosomatid flagellates (Kinetoplastea: Trypanosomatidae) contains a long-known and phylogenetically well-supported species-rich lineage that was provisionally named as the 'jaculum' clade. Its members were found in representatives of several unrelated families of heteropteran bugs captured in South and Central America, Europe, Africa, and Asia. However, this group resisted introduction into the culture, a needed prerequisite for its proper characterization. Here we describe four new cultivable species, which parasitize various parts of their hosts' intestine, including the thoracic and abdominal part of the midgut, hindgut, and Malpighian tubules. Morphologically, the cultured flagellates vary from relatively short stumpy promastigotes to long slender leptomonad cells. Some species form straphangers (cyst-like amastigotes) both in vivo and in vitro, initially attached to the basal part of the flagellum of the mother cell, from which they subsequently detach. To formally classify this enigmatic monophyletic cosmopolitan clade, we erected Obscuromonas gen. nov., including five species: O. modryi sp. nov. (isolated from the true bug host species Riptortus linearis captured in the Philippines), O. volfi sp. nov. (from Catorhintha selector, Curaçao), O. eliasi sp. nov. (from Graptostethus servus, Papua New Guinea), O. oborniki sp. nov. (from Aspilocoryphus unimaculatus, Madagascar), and O. jaculum comb. nov. (from Nepa cinerea, France). Obscuromonas along with the genus Blastocrithidia belongs to the newly established Blastocrithidiinae subfam. nov., (Copyright © 2021 Elsevier GmbH. All rights reserved.)

Girault (1917) very briefly described and diagnosed the encyrtid wasp species Ooencyrtus californicus Girault (Hymenoptera: Encyrtidae) with the following limited data (p. 22): "Sacramento, California, from bug eggs on Pinus sabiniana, September". The first author examined its two syntypes, poorly mounted on a slide, of which only parts of the four antennae and a slightly damaged fore wing remain (Triapitsyn et al. 2020). They concluded that this species was better considered a nomen dubium until fresh specimens could be collected from the same host plant in or near the type locality. In July 2019, an effort was made to re-collect O. californicus from foothill (or gray) pine, Pinus sabiniana (Pinaceae), in the Oakhurst area of Madera County, California, USA, where this pine is abundant, but no specimens were captured. At the same time, a very similar insect was collected from sentinel eggs of the invasive stink bug Bagrada hilaris (Burmeister) in Riverside, California. This insect was cultured in the laboratory as part of a B. hilaris biological control program. Due to the poor preservation of the type specimens and in the absence of genetic evidence, it was impossible to positively and properly attribute specimens of this newly collected parasitoid to O. californicus. Therefore, to provide a much needed scientific name for this native egg parasitoid, it was described as Ooencyrtus lucidus Triapitsyn Ganjisaffar in Triapitsyn et al. (2020). The two nominal species were separated based on minor differences in the proportions of the scape, pedicel, and first funicular segment of the female antenna (Triapitsyn et al. 2020). John S. Noyes, world expert on Encyrtidae and one of the reviewers of Triapitsyn et al. (2020), disagreed with the erection of O. lucidus as a new taxon based on the morphological similarities of the female antenna with that of O. californicus, so the description was published with the understanding that the two might be conspecific. But to settle this one way or another new specimens of O. californicus had to be collected to allow morphological and molecular analyses.

Telenomus podisi Ashmead (Hymenoptera: Scelionidae) is the most important egg parasitoid of Euschistus heros (Fabricius) (Hemiptera: Pentatomidae), and its successful parasitism is related to their searching ability to find suitable hosts under a complex chemical environment using host-reliable cues. Thus, the objective of this study was to elucidate chemical substances on the external layer of E. heros eggs and report its potential kairomonal activity on T. podisi. We tested female wasps in olfactometer system to synthetic compounds obtained from a chemical identification of E. heros egg masses. The synthetic blend was also evaluated in parasitism tests under laboratory and semi-field conditions. We identified 31 substances from egg surface extracts, including monoterpenes, aldehydes and alkanes. Among those compounds, a synthetic solution including camphene, β-pinene, limonene and benzaldehyde-induced chemotaxic behaviour on the wasps and increased the parasitism on E. heros eggs, either in laboratory or semi-field test, suggesting its potential use to T. podisi manipulation and parasitism improvement.

Dispersal determines the spatial dynamics of host-parasite assemblages, particularly during invasions and disease epidemics. The risk of parasitism may create an incentive for dispersal, but infection is expected to reduce dispersal ability, which may alter the host's dispersal response to biotic stressors, including population density. We measured the dispersal of a semiaquatic insect ( Notonecta undulata ) in aquatic mesocosms in which we manipulated the presence of ectoparasitic Hydrachnidia mites and infected conspecifics. We found that parasitism risk increases host dispersal propensity. Using a flight assay, we determined that parasite infection reduces host dispersal ability. Finally, we used a mark-release-recapture study to investigate the joint effects of both parasitism risk and parasite infection on host dispersal in a natural, spatially structured population. We found that parasitism risk reduced dispersal probability, eliminated positive density-dependent dispersal, and increased dispersal distance. Infection had no effect on dispersal in the natural population. Our results show that parasites can both increase and decrease the movement rates of their hosts, depending on the ecological context, and can alter the host's dispersal response to other biotic stressors. Future studies should consider the consequences of this double-headed impact of parasites for landscape connectivity, population persistence, and host-parasite coevolution.

Stink bugs (Hemiptera: Pentatomidae) are agricultural pests of increasing significance in the North Central Region of the United States, posing a threat to major crops such as soybean. Biological control can reduce the need for insecticides to manage these pests, but the parasitism of stink bugs by Tachinidae (Diptera) is poorly characterized in this region. The objective of this study was to evaluate the rate of parasitism of stink bugs by tachinids over 2 yr from nine states across the North Central Region. Parasitism was assessed by quantifying tachinid eggs on the integument of stink bug adults. Parasitism rates (i.e., percent of adult stink bugs with tachinid eggs) were compared across stink bug species, states, stink bug sex, and years. The mean percent parasitism of stink bugs by tachinids was about 6% across the region and did not differ among stink bug species. Mean percent parasitism was significantly higher in Missouri than in northern and western states. In addition, male stink bugs had significantly higher mean percent parasitism than females. Stink bug species commonly found in soybean in the region showed some parasitism and are therefore potentially vulnerable to oviposition by these parasitoids. This is the first study to characterize the level of parasitism of stink bugs by tachinids across the North Central Region., (© The Author(s) 2020. Published by Oxford University Press on behalf of Entomological Society of America.)

Here we describe the new trypanosomatid, Phytomonas borealis sp. n., from the midgut of the spiked shieldbugs, Picromerus bidens (Linnaeus), collected in two locations, Novgorod and Pskov Oblasts of Russia. The phylogenetic analyses, based on the 18S rRNA gene, demonstrated that this flagellate is a sister species to the secondary monoxenous Phytomonas nordicus Frolov et Malysheva, 1993, which was concurrently documented in the same host species in Pskov Oblast. Unlike P. nordicus, which can complete its development (including exit to haemolymph and penetration into salivary glands) in Picromerus bidens, the new species did not form any extraintestinal stages in the host. It also did not produce endomastigotes, indispensable for transmission in other Phytomonas spp. These observations, along with the fact that P. bidens overwinters at the egg stage, led us to the conclusion that the examined infections with P. borealis were non-specific. Strikingly, the flagellates from the Novgorod population contained prokaryotic endosymbionts, whereas the parasites from the second locality were endosymbiont-free. This is a first case documenting presence of intracellular symbiotic bacteria in Phytomonas spp. We suggest that this novel endosymbiotic association arose very recently and did not become obligate yet. Further investigation of P. borealis and its intracellular bacteria may shed light on the origin and early evolution of endosymbiosis in trypanosomatids.

Field and laboratory studies were conducted with the Neotropical red-shouldered stink bug Thyanta perditor (F.) (Heteroptera: Pentatomidae) aiming to evaluate parasitism incidence on adults by tachinid flies (Diptera: Tachinidae), which were raised in the laboratory for identification. Egg deposition by flies on adult body surface was mapped. In addition, nymph and adult incidence on the wild host plant black jack, Bidens pilosa L. (Asteraceae), during the vegetative and the reproductive periods of plant development was studied. Seven species of tachinid flies were obtained: Euthera barbiellini Bezzi (73% of the total) and Trichopoda cf. pictipennis Bigot (16.7%) were the most abundant; the remaining five species, Gymnoclytia sp.; Phasia sp.; Strongygaster sp.; Cylindromyia cf. dorsalis (Wiedemann); and Ectophasiopsis ypiranga Dios & Nihei added 10.3% of the total. Tachinid flies parasitism on T. perditor adults was significantly greater on the dorsal compared to the ventral body surface. On the dorsal surface, the pronotum was significantly preferred and the wings the least preferred site. No differences were observed on the number of tachinid fly eggs deposited on wings, considering the "under" and "above" sites. Results indicated a significantly greater number of nymphs on mature compared to immature seeds. Adults significantly preferred immature compared to mature seeds; both were less abundant on leaves/stems and inflorescences.

Here we characterized the development of the trypanosomatid Blastocrithidia raabei in the dock bug Coreus marginatus using light and electron microscopy. This parasite has been previously reported to occur in the host hemolymph, which is rather typical for dixenous trypanosomatids transmitted to a plant or vertebrate with insect's saliva. In addition, C. marginatus has an unusual organization of the intestine, which makes it refractory to microbial infections: two impassable segments isolate the anterior midgut portion responsible for digestion and absorption from the posterior one containing symbiotic bacteria. Our results refuted the possibility of hemolymph infection, but revealed that the refractory nature of the host provokes very aggressive behavior of the parasite and makes its life cycle more complex, reminiscent of that in some dixenous trypanosomatids. In the pre-barrier midgut portion, the epimastigotes of B. raabei attach to the epithelium and multiply similarly to regular insect trypanosomatids. However, when facing the impassable constricted region, the parasites rampage and either fiercely break through the isolating segments or attack the intestinal epithelium in front of the barrier. The cells of the latter group pass to the basal lamina and accumulate there, causing degradation of the epitheliocytes and thus helping the epimastigotes of the former group to advance posteriorly. In the symbiont-containing post-barrier midgut segment, the parasites either attach to bacterial cells and produce cyst-like amastigotes (CLAs) or infect enterocytes. In the rectum, all epimastigotes attach either to the cuticular lining or to each other and form CLAs. We argue that in addition to the specialized life cycle B. raabei possesses functional cell enhancements important either for the successful passage through the intestinal barriers (enlarged rostrum and well-developed Golgi complex) or as food reserves (vacuoles in the posterior end)., Competing Interests: The authors have declared that no competing interests exist.

Nosema maddoxi Becnel, Solter, Hajek, Huang, Sanscrainte, & Estep, a microsporidian species native to the United States, has been found infecting the invasive brown marmorated stink bug, Halyomorpha halys (Stål). Microsporidian infections in insects often shorten lifespans, decrease fecundity, prolong development, and stunt growth. This study was conducted to determine the impact of N. maddoxi on H. halys fitness. Adult females (2 doses) and nymphs (1 dose) drank suspensions of N. maddoxi spores to promote infection. Adult females receiving a high dose died faster than the controls. Nosema maddoxi infections impacted female egg production and egg viability at both doses compared with the controls. Infections were transmitted to 34.9% of adult males caged with infected females. As the number of days after inoculation increased, infection intensity (# spores found within an infected individual) for both adult treatments transitioned from low-intensity to high-intensity. Infected nymphs died significantly sooner than the controls. Of the treated nymphs, 55.9% died before molting into the fourth instar and only 26.5% eclosed to adults. Nymphal development rate and size were not impacted by N. maddoxi infection. These results indicate that N. maddoxi infection can negatively impact the lifespan of adult females, female fecundity, egg viability, and nymphal survival, which we hypothesize would negatively impact H. halys population densities., (Copyright © 2019 Elsevier Inc. All rights reserved.)

Successful biological control requires detailed knowledge about the mass rearing conditions of the control agents in order to ensure higher quality of field-released insects. Thus, we investigated whether rearing fluctuating thermal condition would affect the fitness and costs of the parasitoid wasp Telenomus podisi Ashmead (a biocontrol agent used for controlling the Neotropical brown stink bug Euschistus heros (Fabricius)) when compared with parasitoid reared at constant temperature condition, which is commonly used in insect facilities. Parasitoids were reared under either constant (continuous exposure at 25 ± 2°C) or fluctuating temperature conditions (i.e., 30 ± 2°C during day and 20 ± 2°C at night) during four consecutive generations. Our results indicated that tested fluctuating temperature is more suitable for rearing of T. podisi as such temperature condition not only resulted in fitness benefits (e.g., shorter developmental time, longer female longevity, higher fecundity/fertility) but also reduced (approximately 23.5%) the estimated costs for producing the parasitoids. Furthermore, rearing T. podisi under fluctuating temperatures improved tolerance to low constant temperatures (i.e., 20°C) without changing the tolerance to constant high temperatures (30°C) in the fourth generation. Surprisingly, even parasitoids that developed under fluctuating thermal conditions performed better than those reared at constant temperature of 25°C. Collectively, our findings suggest that T. podisi reared under fluctuating thermal condition can tolerate better fluctuating temperatures that normally occur both during long periods of transport and in agricultural ecosystems, which will increase the quality and productivity of mass-reared T. podisi for inundative releases.

This study investigates the influence of parasitoid age and egg age of the hosts Euschistus heros (Fabricius) and Dichelops melacanthus (Dallas) on parasitism of Telenomus podisi Ashmead. Six separate bioassays were conducted: parasitism on eggs of E. heros (bioassay 1) and D. melacanthus (bioassay 2) by T. podisi females of different age (1, 5, and 10 days old); parasitism by T. podisi on eggs of different age (1, 2, 3, 4, and 5 days of embryonic development) of the hosts E. heros (bioassay 3) and D. melacanthus (bioassay 4); preference of T. podisi females for eggs at different embryonic developmental stages (eggs of 1, 2, 3, 4, and 5 days) of the hosts E. heros (bioassay 5) and D. melacanthus (bioassay 6). The age of T. podisi females and their hosts affected parasitism on both E. heros and D. melacanthus eggs. Overall, the parasitism rate was higher in older than younger parasitoids, independent of the tested host species, and host eggs between 1 and 3 days old were similarly parasitized. Thus, in T. podisi mass rearing facilities, it is recommended to use older adults (5 to 10 days old) as mother wasps to increase parasitism on the offered eggs. In addition, when hosts are completely absent in the field, or climatic conditions are unfavorable for release, mass-reared adults can be kept in the laboratory (25°C) for up to 10 days for later release in the field without any impairment of their subsequent parasitism performance.

Trypanosomatids are protozoan parasites that infect thousands of globally dispersed hosts, potentially affecting their physiology. Several species of trypanosomatids are commonly found in phytophagous insects. Leptomonas wallacei is a gut-restricted insect trypanosomatid only retrieved from Oncopeltus fasciatus. The insects get infected by coprophagy and transovum transmission of L. wallacei cysts. The main goal of the present study was to investigate the effects of a natural infection by L. wallacei on the hemipteran insect O. fasciatus, by comparing infected and uninfected individuals in a controlled environment. The L. wallacei-infected individuals showed reduced lifespan and morphological alterations. Also, we demonstrated a higher infection burden in females than in males. The infection caused by L. wallacei reduced host reproductive fitness by negatively impacting egg load, oviposition, and eclosion, and promoting an increase in egg reabsorption. Moreover, we associated the egg reabsorption observed in infected females, with a decrease in the intersex gene expression. Finally, we suggest alterations in population dynamics induced by L. wallacei infection using a mathematical model. Collectively, our findings demonstrated that L. wallacei infection negatively affected the physiology of O. fasciatus, which suggests that L. wallacei potentially has a vast ecological impact on host population growth.

Trissolcus japonicus (Ashmead), an Asian parasitoid of Halyomorpha halys (Stål) (Hemiptera: Pentatomidae), was first detected in North America in 2014. Although testing in quarantine facilities as a candidate for classical biological control is ongoing, adventive populations have appeared in multiple sites in the United States, Canada, and Europe. Extensive laboratory testing of T. japonicus against other North American pentatomids and H. halys has revealed a higher rate of parasitism of H. halys, but not complete host specificity. However, laboratory tests are necessarily artificial, in which many host finding and acceptance cues may be circumvented. We offered sentinel egg masses of three native pentatomid (Hemiptera: Pentatomidae) pest species (Chinavia hilaris (Say), Euschistus conspersus Uhler, and Chlorochroa ligata (Say)) in a field paired-host assay in an area with a well-established adventive population of T. japonicus near Vancouver, WA. Overall, 67% of the H. halys egg masses were parasitized by T. japonicus during the 2-yr study. Despite the 'worst case' scenario for a field test (close proximity of the paired egg masses), the rate of parasitism (% eggs producing adult wasps) on all three native species was significantly less (0.4-8%) than that on H. halys eggs (77%). The levels of successful parasitism of T. japonicus of the three species are C. hilaris > E. conspersus > C. ligata. The potential impact of T. japonicus on these pentatomids is probably minimal., (© The Author(s) 2019. Published by Oxford University Press on behalf of Entomological Society of America.)

We describe the monoxenous trypanosomatids parasitizing true bugs and flies on the island of Curaçao. Out of 248 examined true bugs belonging to 17 species, 93 individuals were found to be infected (overall 38% prevalence) by at least one trypanosomatid species (referred to as typing units; TUs). Out of 80 flies, six were infected. All detected trypanosomatids were compared based on their 18S rRNA sequences with TUs parasitizing bugs and flies described from mainland South America, allowing us to assess their diversity and distribution. Besides Leptomonas pyrrhocoris and Leptomonas seymouri, two known species of the subfamily Leishmaniinae, our analysis revealed six new TUs falling into the groups 'jaculum', Blastocrithidia and Herpetomonas. Moreover, two new members of the genus Phytomonas and three new TUs belonging to the monophyletic group designated as 'new clade II' sensu Mol. Phylogenet. Evol, 69, 255 (2013) were isolated. The detected trypanosomatids were characterized by moderate diversity (13 TUs) species richness. Out of nine and four TUs from the heteropteran and dipteran hosts, respectively, 11 TUs have not been encountered before. Although a sampling bias may partially affect the comparison between trypanosomatid communities on Curaçao and the mainland, the high proportion of unique TUs from the former location suggests that the prominent role of islands in increasing the global diversity of macroscopic organisms may also extend to their protistan parasites., (© 2018 International Society of Protistologists.)

This study examined the expression and role of vitellogenin (Vg) in the body of the firebug Pyrrhocoris apterus (Heteroptera, Insecta) during infection elicited by two entomopathogenic organisms, the nematode Steinernema carpocapsae and the fungus Isaria fumosorosea Infection by S. carpocapsae significantly upregulated Vg mRNA expression in the male body. The corresponding increase in Vg protein expression was also confirmed by electrophoretic and immunoblotting analyses. Remarkably, in females, the opposite tendency was noted. Nematodal infection significantly reduced both Vg mRNA and Vg protein expression levels in fat body and hemolymph, respectively. We speculate that infection of reproductive females reduces Vg expression to a level that is still sufficient for defense, but is insufficient for reproduction. This circumstance reduces energy expenditure and helps the individual to cope with the infection. Importantly, purified Vg significantly inhibited growth of Xenorhabdus spp., an entomotoxic bacteria isolated from S. carpocapsae. However, the effect of Vg against I. fumosorosea was not so obvious. The fungus significantly stimulated Vg gene expression in males; however, a similar increase was not recapitulated at the protein level. Nevertheless, in females, both mRNA and protein Vg levels were significantly reduced after the fungal infection. The obtained data demonstrate that Vg is probably an important defense protein, possibly with a specific activity. This considerably expands the known spectrum of Vg functions, as its primary role was thought to be limited to regulating egg development in the female body., Competing Interests: Competing interestsThe authors declare no competing or financial interests., (© 2019. Published by The Company of Biologists Ltd.)

Here we described a new trypanosomatid species, Phytomonas lipae, parasitizing the dock bug Coreus marginatus based on axenic culture and in vivo material. Using light and electron microscopy we characterized the development of this flagellate in the intestine, hemolymph and salivary glands of its insect host. The intestinal promastigotes of Phytomonas lipae do not divide and occur only in the anterior part of the midgut. From there they pass into hemolymph, increasing in size, and then to salivary glands, where they actively proliferate without attachment to the host's epithelium and form infective endomastigotes. We conducted molecular phylogenetic analyses based on 18s rRNA, gGAPDH and HSP83 gene sequences, of which the third marker performed the best in terms of resolving phylogenetic relationships within the genus Phytomonas. Our inference demonstrated rather early origin of the lineage comprising the new species, right after that of P. oxycareni, which represents the earliest known branch within the Phytomonas clade. This allowed us to compare the development of P. lipae and three other Phytomonas spp. in their insect hosts and reconstruct the vectorial part of the life cycle of their common ancestor., Competing Interests: The authors have declared that no competing interests exist.

The brown marmorated stink bug, Halyomorpha halys (Stål) (Hemiptera: Pentatomidae), is an invasive agricultural and nuisance pest that has established across much of the United States and caused significant crop losses in the Mid-Atlantic region. While it has been monitored extensively using ground-deployed pheromone traps, the vertical distribution of its life stages in the canopy of wild tree hosts has not been examined. In Virginia, small pyramid traps baited with 'low-dose' H. halys pheromone lures were deployed via a pulley system at the lower, mid-, and upper canopy of female tree of heaven (Ailanthus altissima (Mill.) Swingle) in 2016 and 2017 and male A. altissima and hackberry (Celtis occidentalis L.) in 2017. Weekly captures of adults and nymphs were recorded throughout each season. Each year, additional female A. altissima trees were felled during the two main periods of H. halys oviposition. The number and relative locations of all pentatomid egg masses found on foliage were recorded and any parasitoids that emerged from them were identified. Halyomorpha halys adults and nymphs were captured in greatest numbers in upper canopy traps and in lowest numbers in traps near the tree base. More H. halys egg masses were collected from mid-canopy than from the lower or upper canopy. The adventive egg parasitoid, Trissolcus japonicus (Ashmead) (Hymenoptera: Scelionidae), emerged most frequently from egg masses found at mid-canopy and was not recovered from those in the lower canopy. Results are discussed in relation to the foraging ecology of H. halys and its natural enemies, including TT. japonicus.

Halyomorpha halys (Stål) (Heteroptera: Pentatomidae), the brown marmorated stink bug (BMSB), is a polyphagous fruit and vegetable pest from eastern Asia and now invasive in the United States and Europe. Earlier research revealed that the main volatile from hexane egg extracts of a western North American pentatomid, Euschistus conspersus Uhler, to which two native scelionid egg parasitoids, Telenomus podisi Ashmead and Trissolcus erugatus Johnson are highly attracted, was methyl (2E,4Z)-2,4-decadienoate, the major component of the conspecific male-produced aggregation pheromone. Conversely, extracts of BMSB eggs lacked the male-produced sesquiterpenoid H. halys pheromone components but contained C16,18,20 aldehydes (hexadecanal, octadecanal, and eicosanal); both egg-surface extracts of BMSB eggs and the C16,18,20 synthetic aldehyde blend repelled the female parasitoids. The goal of the present research was to manipulate egg-surface volatiles to induce the scelionid egg parasitoids to adopt BMSB eggs as a host. Here it was demonstrated that individual females of both parasitoids could be conditioned to H. halys egg-surface aldehydes in Y-tube olfactometer experiments, and this positive memory lasted 3-4 d. Furthermore, conditioned wasps successfully parasitized fresh H. halys eggs in no choice tests, and their offspring continued to successfully parasitize fresh H. halys eggs for 2-3 more generations. Eventually, both the individual and generational memories faded. Unconditioned female parasitoids were not attracted to H. halys egg extract or the synthetic blend of egg-surface aldehydes, nor did they parasitize BMSB eggs. Theoretical and practical implications of the research are discussed in the contexts of Hopkins' host selection principle, biological control, and instinct evolution.

Over the last 50 yr, the geographical distribution of soybean crop production in Brazil has expanded from the southern region to Maranhão state in the north. We evaluated if this latitudinal expansion affected the community of stink bugs (Hemiptera: Pentatomidae) and their parasitoids. The fauna of stink bugs and their adult parasitoids were studied in nine soybean production regions in Brazil. Stink bugs were sampled using a shake cloth and and held in laboratory cages with natural diet to await emergence of parasitoids. Stink bug and parasitoid species composition did not shift along the latitudinal gradient. Euschistus heros (Fabricius, 1798) (Hemiptera: Pentatomidae) was the most abundant stink bug and occurred in all sampling regions. Hexacladia smithii Ashmead, 1891 (Hymenoptera: Encyrtidae), Cylindromyia brasiliana (Townsend, 1927), Ectophasiopsis sp., Eutrichopoda sp., Gymnoclytia sp., Phasia sp., and Trichopoda sp. (Diptera: Tachinidae) were the parasitoid adult stink bugs that we registered. Parasitism indexes were low, ranging from 0.77 to 6.05% through the regions. On the other hand, parasitism rates were higher in areas with reduced insecticide application in comparison to areas with intensive insecticide use.

Lethal and sublethal effects of insecticides used in organic agriculture were tested against Anastatus reduvii and Telenomus podisi, native North American hymenopteran egg parasitoids of the native Euschistus servus Say (Hemiptera: Pentatomidae) and the invasive Halyomorpha halys Stål. Entrust (spinosad), PyGanic (pyrethrin), Neemix (azadirachtin), and Azera (pyrethrin + azadirachtin) were tested at equivalent field rates of 1×, 0.5×, and 0.1×. Bioassays included insecticide exposure to parasitoids through residue on substrate, parasitized host eggs, and their food source. When exposed to dried residues, Entrust caused 100% mortality at the 0.5× rate to both species; PyGanic, Neemix, and Azera exhibited low toxicity. Exposure of parasitized host eggs to Entrust 1× during the egg stage of parasitoid development reduced parasitoid emergence compared to all other treatments in both species. Anastatus reduvii emergence was also reduced by PyGanic at 0.5× and 1×. Parasitoid emergence from host eggs exposed during the pupal stage was more variable than egg stage exposure; emergence of both species was reduced in 0.5× and 1× rates of PyGanic, and A. reduvii was reduced in the 0.5× rate of Entrust compared to controls. Longevity of emerged parasitoids surviving exposure within host eggs showed that Entrust was more deleterious than Neemix or PyGanic. When A. reduvii was fed insecticide-laced honey, all treatments except Neemix at 0.1× reduced adult longevity compared to the control. These studies demonstrated that insecticides commonly used in organic agriculture can negatively affect two common parasitoids of stink bugs; specifically, negative effects were most pronounced with Entrust, and variable with Neemix and Pyganic.

Successful parasitoid rearing is crucial for augmentative biological control. A low temperature preservation protocol allowing the availability of host and parasitoid year-round was evaluated in this study in four bioassays: (1) host eggs [Euschistus heros (Fabricius)] stored at - 196, - 80, and - 20°C for up to 70 days prior to exposure to Telenomus podisi Ashmead parasitism; (2) Euschistus heros eggs removed from storage at - 196°C after 70 days and kept at 5°C for up to 9 days prior to exposure to T. podisi parasitism; (3) Telenomus podisi adult emergence of insects stored as pupae at 5°C; and (4) fitness of adults of T. podisi stored at 5°C. Higher parasitism was observed in parasitoids reared on E. heros eggs stored at - 196 and - 80°C. Host eggs removed from - 196°C and stored at 5°C for up to 6 days did not impact T. podisi parasitism and development. Storage of T. podisi pupae for more than 7 days negatively affected parasitoid biology. Storing T. podisi adults at 5°C for up to 6 days does not alter the biological parameters of the parasitoid. Thus, parasitoids can be stored as pupae or adults as well as its host E. heros eggs. Our findings can be applied to improve the feasibility of year-round insect production.

Salinity is increasing in aquatic ecosystems in the Mediterranean region due to global change, and this is likely to have an important impact on host-parasite interactions. Here we studied the relationships between infection by ectoparasitic water mites and salinity variation, on survival and fecundity of water boatmen Corixidae in the laboratory. Larvae of Sigara lateralis parasitised by larval mites (Hydrachna skorikowi) had lower survivorship, and failed to moult to the adult stage. In adult corixids (S. lateralis and Corixa affinis) fitness was reduced at high salinities and in individuals infected by H. skorikowi, both in terms of survival and fecundity. We also found evidence for parasitism-salinity interactions. Our results suggest that ongoing increases in salinity in Mediterranean ponds due to climate change and water abstraction for agriculture or urban use have a strong impact on water bugs, and that their interactions with ectoparasites may modify salinity effects., Competing Interests: Co-author Andy J. Green serves as an Academic Editor for this journal. This does not alter our adherence to PLOS ONE policies on sharing data and materials.

Stink bugs are important insect pests of soybean, Glycine max (L.), in Louisiana. Scouting, economic thresholds, and insecticide applications are the main strategies used in the integrated pest management of soybean insect pests. However, biological control of stink bug eggs by parasitoids has the potential to reduce populations of these pests. A survey of stink bug egg parasitoids was conducted at different sites in Louisiana from 2008 to 2010. Similarly, a study on the incidence of stink bug egg parasitoids within soybean vertical strata and plant structures was conducted in 2009 to 2011. Species of stink bug eggs collected during the studies included the redbanded stink bug, Piezodorus guildinii (Westwood), the brown stink bug species complex, Euschistus spp., the southern green stink bug, Nezara viridula L., the spined soldier bug, Podisus maculiventris (Say), and the green stink bug, Chinavia hilaris (Say). The percentage of stink bug eggs parasitized in the central location of Louisiana was higher compared with the northwest location during the 2008 to 2010 growing seasons. There were no differences in percentage of stink bug egg parasitism within soybean plant strata between 2009 and 2011. Telenomus podisi Ashmead (Hymenoptera: Platygastridae) was the most abundant parasitoid, and it showed preference for P. guildinii. Other platygastrid species that emerged included Trissolcus euschisti Ashmead (Hymenoptera: Platygastridae), Gryon obesum Masner (Hymenoptera: Platygastridae), and Telenomus longicornis Johnson (Hymenoptera: Platygastridae). This study is the first to report the parasitization of P. guildinii eggs on soybeans in the United States.

Parasitoid host selection is mainly mediated by chemical cues, which can be adjusted by experience, changing their innate behavior. Therefore, this study evaluated if immature experience (pre-imaginal conditioning) on eggs and volatiles from different host eggs has influence on parasitism and chemotaxic behavior of Telenomus podisi Ashmead and/or Trissolcus basalis Wollaston (Hymenoptera: Scelionidae). Both wasp species were submitted to a multiple-choice parasitism test among Euschistus heros (Fabricius), Piezodorus guildinii (Westwood), and Nezara viridula L. (Hemiptera: Pentatomidae) (Hemiptera: Pentatomidae) egg masses. Eggs from these three stink bugs were equally offered to female parasitoids. After that, adults which emerged from each host were also exposed to parasitism in a multiple-choice test for up to an additional generation. Moreover, in olfactometer "Y," the behavior of innate and experienced T. podisi females to volatiles from hosts' egg extracts was tested, to study their learning and memory ability. The original host had influence on T. podisi parasitism; however, T. basalis always parasitized more N. viridula eggs independently of its last rearing host. Innate T. podisi females responded positively to E. heros and P. guildinii egg volatiles, but this behavior was not observed in N. viridula. When T. podisi females were experienced on egg volatiles from a new host, they showed significant learning and memory ability for the specific host volatile for, at least, 24 h. Experienced wasps responded positively to N. viridula and through this result we have evidences about the possibility to manipulate wasp's preferences to a specific target host.

The brown marmorated stink bug, Halyomorpha halys (Stål) (Hemiptera: Pentatomidae), has spread across North America and is causing serious economic damages. Current management of this pest is based primarily on use of insecticides, which can disrupt integrated pest management programs. Alternatively, biological control is a more benign management tactic. This study provides the first examination of potential impact of parasitoids and predators of pentatomid eggs in Minnesota. Over 2 yr, 10,074 fresh and 9,870 frozen H. halys eggs were deployed in two forest and two soybean habitats in St. Paul, Minnesota from June to August. Our results demonstrate that rates of parasitism and predation were low, accounting for only 0.4 and 3.7%, respectively, across years, habitats, and egg states. In general, the parasitoid Telenomus podisi Ashmead (Hymenoptera: Scelionidae), had higher impacts on H. halys eggs in soybean, and generalist predators were more prevalent in forest habitats. Overall, predation was higher on fresh versus frozen eggs, and parasitism was not consistent across egg states. Although the rates of H. halys mortality due to natural enemies were low, results of our study may be conservative estimates of their true impact. Also, sentinel egg mass surveys should account for undeveloped parasitoids to better quantify H. halys egg mortality by native parasitoids. Alternative management tactics, such as the introduction of Trissolcus japonicus (Ashmead) (Hymenoptera: Scelionidae), could be considered to improve biological control of H. halys. Our findings serve as the foundation for future work on biological control of this pest and other pentatomids.

Successful biological control requires detailed knowledge about host preferences of the released parasitoid, because the presence of alternative hosts may affect the control of the target pest. The objective of this work was therefore to evaluate host preferences of Telenomus podisi Ashmead among the eggs of three stink bug species: Dichelops melacanthus Dallas, Euschistus heros Fabricius, and Podisus nigrispinus Dallas (Heteroptera: Pentatomidae). Three independent experiments were carried out to study host preferences among the following: (1) E. heros, D. melacanthus dallas, and P. nigrispinus (bioassay 1); (2) E. heros and D. melacanthus (bioassay 2); and (3) D. melacanthus and P. nigrispinus (bioassay 3). A single bioassay (bioassay 4) was carried out to evaluate the egg size of E. heros, D. melacanthus, and P. nigrispinus. Two more bioassays were carried out: bioassay 5 to study the biological characteristics of T. podisi reared on E. heros, D. melacanthus, and P. nigrispinus eggs, and bioassay 6 to study the morphological characters of T. podisi reared on those different host eggs. Overall, T. podisi consistently preferred eggs of D. melacanthus to those of the other studied hosts, due to probably their better nutritional value; hypothesis that is supported by the fast T. podisi development and bigger parasitoids when reared on D. melacanthus and P. nigrispinus eggs. This allows suggesting that neither pre-imaginal conditioning nor associative learning nor α-conditioning are relevant to T. podisi parasitism. Thus, E. heros eggs could be successfully used for mass rearing of this parasitoid for successive generations since it would not affect its parasitism on other species of the Pentatomidae family in the field.

Psix saccharicola (Mani) (Hymenoptera: Platygastridae) is a solitary egg parasitoid of the pistachio green stink bug, Acrosternum arabicum (Wagner) (Hemiptera: Pentatomidae), which is one of the most important pests of pistachio in Iran. Augmentation of P. saccharicola field populations using mass-reared individuals may provide an alternative to conventional pesticide use for pistachio green stink bug control. Cold storage is an important component of mass-rearing protocols for optimum timing of host egg parasitization and potentially extended storage of P. saccharicola pupae prior to adult emergence. The impact of cold storage on A. arabicum eggs for various time intervals at 4.0°C was investigated. Results indicated that host eggs stored at 4.0°C for up to 60 d could be exploited by P. sacchricola, whereas no offspring were produced when eggs were stored for 120 d. The emergence rates of the F1 and F2 generations declined with increased host egg storage time. Both sex ratio and survival rate of the F2 generation decreased as the refrigeration time of host eggs increased. The impact of cold storage on P. saccharicola pupae was evaluated. Reared pupae of P. saccharicola were held for 1 wk at three temperatures and compared with a control (27 ± 1°C). Psix saccharicola pupae were tolerant to cold storage at 8 and 12°C. Cold storage adversely affected mean adult emergence at 4°C, which decreased following low temperature exposure. Furthermore, mean percentage survivorship was unaffected by storage at low temperatures in the F1 generation, but was reduced at 4°C. The sex ratio of the F1 generation became more male-biased when held at lower storage temperatures. The highest female proportion was observed at 12°C.

A strain (ARP14) of the entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin was isolated from field-collected Riptortus pedestris (Fabricius) (Hemiptera: Alydidae). The lethal median concentration of the ARP14 strain was compared with that of a commercialized strain (GHA) of the same fungus against R. pedestris and its two egg parasitoids, Ooencyrtus nezarae Ishii (Hymenoptera: Encyrtidae) and Gryon japonicum (Ashmead) (Hymenoptera: Platygastridae). Mortality and mycosis rates were evaluated after exposure to five concentrations of the fungus, i.e., 1×109, 1×108, 1×107, 1×106, and 1×105 conidia/mL, using a glass scintillation vial as an exposure arena in 25.0 ± 0.5°C and 93.7 ± 2.9% RH. The lethal median concentrations (LC50) for 2nd and 4th instar nymphs, and adults of R. pedestris were not significantly different between the two strains of B. bassiana. However, the mycosis rate of ARP14 was 1.3 and 1.8 times higher than that of the GHA strain in 4th instar nymphs and adult females of R. pedestris, respectively, at the 1×108 conidia/mL concentration. More interestingly, the mycosis rates at 1×108 conidia/mL concentration in the parasitoids G. japonicum and O. nezarae were much lower in the ARP14 strain (15.0 and 0%) than in the GHA strain (73.3 and 66.0%), respectively, suggesting that the B. bassiana strain ARP14 is less virulent to these parasitoids than the commercially available strain. Our results suggest that B. bassiana ARP14 may be a potential new biopesticide against R. pedestris with fewer negative effects on beneficial parasitoids than currently available options.

Trissolcus japonicus (Ashmead) (Hymenoptera: Scelionidae) is an egg parasitoid wasp from East Asia. It is considered to be an important natural enemy of the brown marmorated stink bug, Halyomorpha halys (Stål) (Hemiptera: Pentatomidae). Ongoing efforts to evaluate the physiological host range of this potential classical biological control agent have demonstrated that the wasp can parasitize at least 11 genera of nontarget Pentatomoidea in North America in no-choice tests. However, factors such as host-acceptance behavior that might affect nontarget parasitism in the field are poorly understood. We evaluated the preference of T. japonicus females provided with egg clusters of H. halys, Thyanta custator accerra McAtee (Hemiptera: Pentatomidae), Podisus maculiventris (Say) (Hemiptera: Pentatomidae), and Euschistus variolarius (Palisot de Beauvois) (Hemiptera: Pentatomidae) when wasps were reared on the target pest, H. halys, and the nontarget species, T.c. accerra and P. maculiventris. When reared on H. halys, T. japonicus showed a strong preference for H. halys, and frequently rejected nontarget hosts after inspection. When T. japonicus was reared on nontarget hosts, females showed reduced host-specificity, but were significantly smaller and produced a lower number of offspring. These results suggest that innate host-acceptance behavior is likely to reduce parasitism on some nontarget Pentatomoidea which T. japonicus will encounter in the field. Future efforts should be made to examine the ability of the wasp to reproduce long term on nontarget species when H. halys eggs are limited.

Since 2014, populations of the kudzu bug, Megacopta cribraria (F.) (Hemiptera: Plataspidae), have declined in the southeastern United States and seldom require treatment. This decline follows the discovery of Paratelenomus saccharalis (Dodd; Hymenoptera: Platygastridae), a non-native egg parasitoid. The objective of this project was to observe the temporal and spatial dynamics of P. saccharalis parasitism of kudzu bug egg masses in commercial soybean fields. Four fields were sampled weekly for kudzu bugs and egg masses at a density of one sample per 0.6 ha. Sampling commenced when soybean reached the R2 maturity stage and continued until no more egg masses were present. Responses including kudzu bugs, egg masses, and parasitism rates were analyzed using ANOVA, Spatial Analysis by Distance Indices (SADIE), and SaTScan spatial analysis software. Egg masses were collected from the field, held in the lab and monitored for emergence of kudzu bug nymphs or P. saccharalis. Kudzu bug populations were generally lower than previously reported in the literature and spatial aggregation was not consistently observed. Egg parasitism was first detected in early July and increased to nearly 40% in mid-August. Significant spatial patterns in parasitism were observed with spatio-temporal clusters being loosely associated with clusters of egg masses. There were no significant differences in parasitism rates between field margins and interiors, suggesting that P. saccharalis is an effective parasitoid of kudzu bug egg masses on a whole-field scale., (© The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Squash bug, Anasa tristis DeGeer (Hemiptera: Coreidae), is a major pest of squash and pumpkins in the United States. In order to better understand the importance of natural egg parasitism of this species in Virginia, we conducted a 2-yr statewide survey. In total, 1,127 squash bug egg masses (~20,000 total eggs) were sampled from squash and pumpkins from 43 counties in Virginia from 2014 to 2015. Egg masses were brought back to the lab to record levels of squash bug nymphal emergence or adult parasitoid eclosion and identification. Over 50% of the total squash bug eggs collected statewide were parasitized. Gryon pennsylvanicum Ashmead (Hymenoptera: Scelionidae) was the predominant egg parasitoid accounting for over 98% of all parasitoid adults recovered. The only other species emerging from squash bug eggs was Anastatus reduvii Howard (Hymenoptera: Eupelmidae), which is a generalist parasitoid. G. pennsylvanicum was found in 75% of the counties surveyed with the highest levels of parasitism occurring in the Northern, Southwestern Mountain, and Western Piedmont regions of the state and the lowest levels of parasitism occurring in the Tidewater region in the southeastern portion of the state. Based on this 2-yr survey, G. pennsylvanicum was determined to be a major natural enemy of squash bug, significantly reducing the number of nymphs that emerge from deposited eggs. Conservation of this natural enemy should therefore be a priority for integrated pest management programs in cucurbits., (© The Author(s) 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

Stylostomes (feeding tubes) of Hirsutiella zachvatkini (Schluger) (Trombiculidae), feeding on bank voles [Myodes glareolus (Schreber)], and of Trombidium holosericeum (L.) (Trombidiidae), feeding on larvae of Stenodemini sp. (Heteroptera, Miridae), were studied by TEM methods and on semi-thin sections. The stylostome of H. zachvatkini is a homogeneous structure of low electron density and without strict margins. It extends within the concave host epidermis, undergoing hyperplasia and hyperkeratosis. TEM does not reveal any obvious stratification in the stylostome walls. The cheliceral movable digits are moved apart by 5-6 µm and tightly applied/adhered to the stylostome substance. A local area beneath the open end of the stylostome canal is not empty but contains a nearly homogeneous substrate, which can pass into the central stylostome canal. The latter is mostly free of contents. In contrast to H. zachvatkini, larvae of T. holosericeum form a root-like stylostome chaotically branching within the clear space underneath the host cuticle free of tissue elements. Tubules of the distal stylostome branches become progressively thinner and disappear blindly. As in H. zachvatkini, the stylostome walls of T. holosericeum are devoid of stratification but show moderate to high electron density. The cheliceral movable digits are moved apart by the same distance, as in H. zachvatkini, and tightly applied to the stylostome substance. The lumen of the central canal is either electron lucent, in the distal portions, or filled with a fine granular or homogeneous substrate of low electron density in the proximal portions forming a type of ampoule. This study shows that Trombiculidae and Trombidiidae share similar initial stages of stylostome formation but the resultant stylostome of each family is distinctly different.

We conducted a field study to determine seasonal egg parasitism rates of the kudzu bug Megacopta cribraria (F.) on the kudzu plant, Pueraria montana (Lour.) Merr. var. lobata (Willd.) Maesen et Almeida ex Sanjappa and Pradeep, in Tokyo, Japan, during the period from May 2014 to September 2014. The eggs of M. cribraria per 1 m2 of kudzu at four locations in Tokyo were collected weekly and parasitism rates were assessed. Eggs of M. cribraria were laid on the kudzu plant from May to September. Megacopta cribraria eggs were parasitized by two parasitoid species, Paratelenomus saccharalis (Dodd) and Ooencyrtus nezarae Ishii. Paratelenomus saccharalis first appeared in May, and its parasitism rates peaked in July and September. Ooencyrtus nezarae first appeared in June and its parasitism rates peaked in July. Except for one location which could not be statistically analyzed because of the small sample size, occurrence of parasitism by P. saccharalis and O. nezarae in M. cribraria egg masses was independent at one location and positively associated at two locations, suggesting that the use of host egg masses by P. saccharalis and O. nezarae is not mutually exclusive. Parasitism rates by P. saccharalis and O. nezarae were significantly lower for egg masses parasitized by both species than for those parasitized by a single species. The proportion of males among O. nezarae progeny was significantly higher for egg masses parasitized by O. nezarae together with P. saccharalis than for those parasitized by O. nezarae alone. These results suggest that parasitism of host egg masses by the two species is influenced by their interspecific interactions., (© The Authors 2017. Published by Oxford University Press on behalf of Entomological Society of America. All rights reserved. For Permissions, please email: journals.permissions@oup.com.)

The role of adipokinetic hormone (AKH) in the firebug Pyrrhocoris apterus adults infected by the entomopathogenic nematode (EPN) Steinernema carpocapsae was examined in this study. It was found that co-application of EPN and AKH enhanced firebug mortality about 2.5 times within 24h (from 20 to 51% in EPN vs. EPN+AKH treatments), and resulted in metabolism intensification, as carbon dioxide production in firebugs increased about 2.1 and 1.6times compared to control- and EPN-treated insects, respectively. Accordingly, firebugs with reduced expression of AKH receptors showed a significantly lower mortality (by 1.6 to 2.9-folds), and lower general metabolism after EPN+AKH treatments. In addition, EPN application increased Akh gene expression in the corpora cardiaca (1.6times), AKH level in the corpora cardiaca (1.3times) and haemolymph (1.7times), and lipid and carbohydrate amounts in the haemolymph. Thus, the outcomes of the present study demonstrate involvement of AKH into the anti-stress reaction elicited by the nematobacterial infection. The exact mechanism by which AKH acts is unknown, but results suggested that the increase of metabolism and nutrient amounts in haemolymph might play a role., (Copyright © 2017 Elsevier Ltd. All rights reserved.)

Phytomonas spp. (phytomonads) are a diverse and globally distributed group of unicellular eukaryotes that parasitize a wide range of plants and are transmitted by insect hosts. Here we report the discovery and characterisation of a new species of Phytomonas, named Phytomonas oxycareni n. sp., which was obtained from the salivary glands of the invasive species of true bug Oxycarenus lavaterae (Heteroptera). The new Phytomonas species exhibits a long slender promastigote morphology and can be found both within the lumen of the insect host's salivary glands as well as within the cells of the salivary gland itself. Sampling multiple individuals from the same population post-winter hibernation on two consecutive years revealed that infection was persistent over time. Finally, phylogenetic analyses of small subunit ribosomal RNA genes revealed that this species is sister to other species within the genus Phytomonas, providing new insight into the evolutionary history of the clade., (Copyright © 2016 Elsevier GmbH. All rights reserved.)

Blastocrithidia papi sp. n. is a cyst-forming trypanosomatid parasitizing firebugs (Pyrrhocoris apterus). It is a member of the Blastocrithidia clade and a very close relative of B. largi, to which it is almost identical through its SSU rRNA gene sequence. However, considering the SL RNA gene these two species represent quite distinct, not even related typing units. Morphological analysis of the new species revealed peculiar or even unique features, which may be useful for future taxonomic revision of the genus Blastocrithidia. These include a breach in the microtubular corset of rostrum at the site of contact with the flagellum, absence of desmosomes between flagellum and rostrum, large transparent vacuole near the flagellar pocket, and multiple vacuoles with fibrous content in the posterior portion of the cell. The study of the flagellates' behavior in the host intestine revealed that they may attach both to microvilli of enterocytes using swollen flagellar tip and to extracellular membranes layers using hemidesmosomes of flagellum. Laboratory experiments on B. papi transmission in P. apterus demonstrated that the parasite may be transmitted vertically (via contaminated surface of eggs) and horizontally (via contaminated substrate and/or necrophagy). We argue that the parasite exploits transmission mechanisms intended for obligate bacterial symbionts of P. apterus., (Copyright © 2016 Elsevier GmbH. All rights reserved.)

A pleiotropic response to the calpain inhibitor MDL28170 was detected in the tomato parasite Phytomonas serpens. Ultrastructural studies revealed that MDL28170 caused mitochondrial swelling, shortening of flagellum and disruption of trans Golgi network. This effect was correlated to the inhibition in processing of cruzipain-like molecules, which presented an increase in expression paralleled by decreased proteolytic activity. Concomitantly, a calcium-dependent cysteine peptidase was detected in the parasite extract, the activity of which was repressed by pre-incubation of parasites with MDL28170. Flow cytometry and Western blotting analyses revealed the differential expression of calpain-like proteins (CALPs) in response to the pre-incubation of parasites with the MDL28170, and confocal fluorescence microscopy confirmed their surface location. The interaction of promastigotes with explanted salivary glands of the insect Oncopeltus fasciatus was reduced when parasites were pre-treated with MDL28170, which was correlated to reduced levels of surface cruzipain-like and gp63-like molecules. Treatment of parasites with anti-Drosophila melanogaster (Dm) calpain antibody also decreased the adhesion process. Additionally, parasites recovered from the interaction process presented higher levels of surface cruzipain-like and gp63-like molecules, with similar levels of CALPs cross-reactive to anti-Dm-calpain antibody. The results confirm the importance of exploring the use of calpain inhibitors in studying parasites' physiology.

We conducted a survey to examine parasitism, predation, and the species composition of native parasitoids attacking sentinel egg masses of the invasive pest, the brown marmorated stink bug, Halyomorpha halys (Stål), in bell pepper, squash, and tomato. A 2-yr survey was conducted for each crop. The species composition of parasitoids was determined for parasitoids that successfully emerged from eggs, parasitoids that were identified from dissections, and parasitoids that were found attending egg masses at the time of collection. The overall rate of brown marmorated stink bug nymphal emergence was 82.7% from laboratory-reared control egg masses compared with only 23.4% from sentinel egg masses due to a combination of predation, parasitism, and unknown mortality causes. Overall, 12.4% of sentinel eggs were parasitized. Parasitoids emerged successfully from only 2.3% of eggs, but they were dissected from an additional 10.1% of eggs. Telenomus podisi Ashmead was the predominant species identified from emerged parasitoids (57.4%), dissected parasitoids (90.2%), and parasitoids attending egg masses (77%). This study provides further evidence that the impact of native parasitoids on brown marmorated stink bug egg mortality may be greatly underestimated based solely on parasitoid emergence rates alone., (Published by Oxford University Press on behalf of Entomological Society of America 2016. This work is written by US Government employees and is in the public domain in the United States.)

A viral genome was assembled de novo from next-generation sequencing (NGS) data from bean bugs, Riptortus pedestris, infected with an entomopathogenic fungus, Beauveria bassiana (Bb), and was further confirmed via the RACE method. This is a novel insect positive-sense single-stranded RNA virus, which we named Riptortus pedestris virus-1 (RiPV-1) (GenBank accession no. KU958718). The genome of RiPV-1 consists of 10,554 nucleotides (nt), excluding the poly(A) tail, which contains a single large open reading frame (ORF) of 10,371 nt encoding a polyprotein (3456 aa) and flanked by 71 and 112 nt at the 5' and 3' untranslated regions (UTR), respectively. RiPV-1 genome organization from the 5' end contains a consensus organization of picorna-like RNA virus helicase, cysteine protease, and RNA-dependent RNA polymerase (RdRp), in addition to two putative structural proteins located at the 3' region and a poly(A) tail at the 3' end. The viral particles were approximately 30nm in diameter with some dispersal distinctive surface projections. Based on the phylogenetic analysis of the RdRp sequences, RiPV-1 was clustered in the unassigned insect RNA viruses with two other viruses, APV and KFV. These three viruses were suggested to constitute a new group of insect RNA viruses. RiPV-1 could be found in all stages of lab-reared bean bugs and was detected abundantly in the thorax, abdomen, midgut and fat body, but not in the reproductive organs and muscle. Interestingly, RiPV-1 replication was increased dramatically in bean bugs 2-6days after fungal infection. In conclusion, a novel insect RNA virus was found by NGS data assembly. This virus can provide further insight into the interaction between virus, fungus and the host., (Copyright © 2016 Elsevier Inc. All rights reserved.)