The First Records of Trissolcus japonicus (Ashmead) and Trissolcus mitsukurii (Ashmead) (Hymenoptera, Scelionidae), Alien Egg Parasitoids of Halyomorpha halys (Stål) (Hemiptera, Pentatomidae) in Serbia.

Serbia has recently begun facing a serious problem with the Brown Marmorated Stink Bug, Halyomorpha halys (Stål), which was first recorded in October 2015. This species belongs to the Pentomidae family and is notorious for causing extensive damage to plants. During the winter, it tends to gather in urban areas, such as houses and different man-made facilities, which has raised concerns among producers and citizens. The population of this species has rapidly increased, causing significant economic damage to cultivated plants. However, despite the alarming situation no natural enemies have yet been identified in Serbia. Therefore, research in 2022 was focused on collecting stink bug eggs to investigate the presence of egg parasitoids. The study identified two foreign Hymenoptera species for the European region, Trissolcus japonicus (Ashmead) and Tr. mitsukurii (Ashmead) (Scelionidae), recorded for the first time in Serbia. Additionally, the list of egg parasitoid species belonging to the Hymenoptera order includes seven local species: Anastatus bifasciatus (Geoffroy), from the Eupelmidae family; Ooencyrtus sp., from the Encyrtidae family; and Telenomus turesis (Walker), Tr. basalis (Wollaston), Tr. belenus (Walker), Tr. colemani (Crawford), and Tr. semistriatus (Nees von Esenbeck), from the Scelionidae family. In total, nine egg parasitoid species were, for the first time, reported as parasitizing H. halys and related species in Serbia.

First Detection of the Adventive Egg Parasitoid of Halyomorpha halys (Stål) (Hemiptera: Pentatomidae) Trissolcus mitsukurii (Ashmead) (Hymenoptera: Scelionidae) in France.

We report the first detection of Trissolcus mitsukurii in France. More than 1860 sentinel egg masses of Halyomorpha halys (BMSB) were exposed in the field during the 2018-2020 period, and 12 specimens of T. mitsukurii emerged from one egg mass. Their taxonomic identification was confirmed both by morphological and molecular analysis. Trissolcus mitsukurii, similar to T. japonicus, is an egg parasitoid of BMSB in its area of origin in Asia, and both species are considered to be candidates for a classical biological control strategy against BMSB. Trissolcus mitsukurii was previously recorded in Italy where it is well established and widespread, and this may be the source of the French population. Possible permanent establishment and dispersion of T. mitsukurii in France should be monitored with emphasis on its potential effect on BMSB populations.

Trissolcus kozlovi in North Italy: Host Specificity and Augmentative Releases against Halyomorpha halys in Hazelnut Orchards.

Trissolcus kozlovi (Hymenoptera: Scelionidae) emerged from field-laid eggs of Halyomorpha halys (Hemiptera: Pentatomidae) in North Italy, and it emerged in significantly higher numbers from fresh H. halys eggs compared to other native scelionids. Since few data on T. kozlovi are available, its host-specificity and some biological traits were investigated in laboratory tests, and its impact after augmentative releases was evaluated in two hazelnut orchards. Among the 12 tested bug species (Hemiptera: Pentatomidae, Scutelleridae), only Nezara viridula was an unsuitable host, while the highest offspring proportions were obtained from Arma custos, Pentatoma rufipes, and Peribalus strictus, followed by Acrosternum heegeri and Palomena prasina. Furthermore, when reared on P. strictus, T. kozlovi showed a high longevity as well as a high adaptation to H. halys eggs. In both hazelnut orchards, T. kozlovi emerged from H. halys eggs after field releases, but it was not found in the next two years. The physiological host range of T. kozlovi was quite similar to that of T. japonicus, and probably T. kozlovi has just begun to attack H. halys as a new host. This aspect needs to be further investigated, as well as its favorable environmental conditions, its distribution and also its possible interaction with T. japonicus, currently present in Italy.

Refining Pheromone Lures for the Invasive Halyomorpha halys (Hemiptera: Pentatomidae) Through Collaborative Trials in the United States and Europe.

Brown marmorated stink bug, Halyomorpha halys, is native to Asia and has invaded North America and Europe inflicting serious agricultural damage to specialty and row crops. Tools to monitor the spread of H. halys include traps baited with the two-component aggregation pheromone (PHER), (3S,6S,7R,10S)-10,11-epoxy-1-bisabolen-3-ol and (3R,6S,7R,10S)-10,11-epoxy-1-bisabolen-3-ol, and pheromone synergist, methyl (2E,4E,6Z)-decatrienoate (MDT). Here, an international team of researchers conducted trials aimed at evaluating prototype commercial lures for H. halys to establish relative attractiveness of: 1) low and high loading rates of PHER and MDT for monitoring tools and attract and kill tactics; 2) polyethylene lure delivery substrates; and 3) the inclusion of ethyl (2E,4E,6Z)-decatrieonate (EDT), a compound that enhances captures when combined with PHER in lures. In general, PHER loading rate had a greater impact on overall trap captures compared with loading of MDT, but reductions in PHER loading and accompanying lower trap captures could be offset by increasing loading of MDT. As MDT is less expensive to produce, these findings enable reduced production costs. Traps baited with lures containing PHER and EDT resulted in numerically increased captures when EDT was loaded at a high rate, but captures were not significantly greater than those traps baited with lures containing standard PHER and MDT. Experimental polyethylene vial dispensers did not outperform standard lure dispensers; trap captures were significantly lower in most cases. Ultimately, these results will enable refinement of commercially available lures for H. halys to balance attraction and sensitivity with production cost.

First Multi-Target Application of Exclusion Net in Nectarine Orchards: Effectiveness against Pests and Impact on Beneficial Arthropods, Postharvest Rots and Fruit Quality.

Over the past few years, there has been an increasing interest in the development of alternative pest control strategies to reduce environmental impact. In this contest, exclusion nets have been evaluated as a sustainable alternative to pesticides. In this study, the use of a photoselective exclusion net was investigated in semi-field conditions as a potential strategy to protect nectarine orchards from different pests (i.e., fruit moths, Halyomorpha halys and Drosophila suzukii) in NW Italy. The presence and abundance of pest populations inside and outside the net, as well as the damage they caused on fruits, were evaluated. Moreover, any possible effects of the net on beneficial arthropods, postharvest rots and fruit quality and nutraceutical parameters were considered. The exclusion net significantly reduced pest populations. At harvest, fruit damage caused by Grapholita molesta and H. halys in netted plots was reduced up to 90% and to 78%, respectively, compared with insecticide-treated plots. The exclusion net allowed the production of healthier fruits with a strong reduction of insecticide treatments (up to seven less) and of their related costs without any negative impact on postharvest rots, neither fruit quality nor nutraceutical properties.

Assessing the Distribution of Exotic Egg Parasitoids of Halyomorpha halys in Europe with a Large-Scale Monitoring Program.

The brown marmorated stink bug Halyomorpha halys is an invasive agricultural pest with a worldwide distribution. Classical biological control has been identified as the most promising method to reduce the populations of H. halys. Adventive populations of two candidates for releases, Trissolcus japonicus and Trissolcus mitsukurii, have recently been detected in Europe. To assess their distribution and abundance, a large-scale survey was performed. From May to September 2019, a wide area covering northern Italy and parts of Switzerland was surveyed, highlighting the expanding distribution of both Tr. japonicus and Tr. mitsukurii. Within four years after their first detection in Europe, both species have rapidly spread into all types of habitats where H. halys is present, showing a wide distribution and continuous expansion. Both exotic Trissolcus showed high levels of parasitism rate towards H. halys, while parasitization of non-target species was a rare event. The generalist Anastatus bifasciatus was the predominant native parasitoid of H. halys, while the emergence of native scelionids from H. halys eggs was rarely observed. The presence of the hyperparasitoid Acroclisoides sinicus was also recorded. This study provided fundamental data that supported the development of the first inoculative release program of Tr. japonicus in Europe.

First Insights on Early Host Plants and Dispersal Behavior of Halyomorpha halys (Hemiptera: Pentatomidae) from Overwintering to Crop Colonization.

Following its first detection in North Italy in 2012, H. halys has become a serious threat in many crops, including hazelnut. The present study aimed at investigating dispersal capacity and behavior in relation to host plants of overwintered adults of H. halys before the colonization of hazelnut crop. Research was carried out in four polyculture areas (from 14 to 50 ha) in north-western Italy in 2018, by using (i) pheromone-baited traps, (ii) visual inspection and beating sheet sampling, and (iii) immunomarking-capture technique. The relative abundance of H. halys was similar between and within the study areas, and the early attractiveness of lures to adults after overwintering was confirmed; the host plants near a pheromone trap (less than 5 m) hosted higher numbers of H. halys than the same plant species far away. Hybrid plane, European spindletree, walnut, oak, and European elder were the first plants on which adult bugs were observed to feed. By immunomarking-capture technique, H. halys showed both short- and long-range dispersal from overwintering sites and/or early host plants to wild and crop plants. Marked adults were found in all zones of each area, irrespective of the distance from the protein treatment. Therefore, H. halys movement patterns depend on the ecosystem features, and plant host distribution and availability. This knowledge together with the interaction between pheromone and early host plants might contribute to the management of post-overwintering adults.

An Insight into the Role of Trissolcus mitsukurii as Biological Control Agent of Halyomorpha halys in Northeastern Italy.

Sustainable strategies such as classical or augmentative biological control are currently being evaluated for the long-term management of the alien invasive pest Halyomorpha halys (Stål) (Hemiptera: Pentatomidae). A three-year study carried out in northeastern Italy was performed to investigate the distribution and field performance of the H. halys egg parasitoid Trissolcus mitsukurii (Ashmead) (Hymenoptera: Scelionidae), in comparison with other parasitoid species. In the study area, adventive populations of T. mitsukurii were present since 2016, representing the earliest detection of this species in Europe. Trissolcus mitsukurii was the most abundant parasitoid and showed a higher "parasitoid impact" (i.e., number of parasitized eggs over the total number of field-collected eggs) compared to the other species, i.e., Anastatus bifasciatus (Geoffroy) (Hymenoptera: Eupelmidae), Trissolcus basalis (Wollaston) and Trissolcus kozlovi Rjachovskij (Hymenoptera: Scelionidae). The hyperparasitoid Acroclisoides sinicus (Huang and Liao) (Hymenoptera: Pteromalidae) was also recorded. Phylogenetic analysis of T. mitsukurii population distinguished two clades, one covering samples from Italy, Japan and China, the other from South Korea. The present study provides promising results for the biological control of a pest that is having a dramatic impact on a wide range of crops worldwide.

The omnivorous predator Macrolophus pygmaeus, a good candidate for the control of both greenhouse whitefly and poinsettia thrips on gerbera plants.

The poinsettia thrips Echinothrips americanus Morgan is a relatively new pest that has spread rapidly worldwide and causes serious damage in both vegetable and ornamental plants. In this study, we investigated if and how effective this pest can be controlled in gerbera by the omnivorous predator Macrolophus pygmaeus (Rambur). Because herbivores on plants can interact through a shared predator, we also investigated how poinsettia thrips control is affected by the presence of the greenhouse whitefly Trialeurodes vaporariorum (Westwood), a pest that commonly coexists with E. americanus in gerbera. In laboratory studies, we found that the predator M. pygmaeus fed on both pests when offered together. Olfactometer tests showed a clear preference of the predators for plants infested by whiteflies but not by thrips. In a greenhouse experiment, densities of both pests on single gerbera plants were reduced to very low levels by the predator, either with both pests present together or alone. Hence, predator-mediated effects between whiteflies and thrips played only a minor role. The plant feeding of the shared predator probably reduced the dependence of predator survival and reproduction on the densities of the two pests, thereby weakening potential predator-mediated effects. Thus, M. pygmaeus is a good candidate for biological control of both pests in gerbera. However, further research is needed to investigate pest control at larger scales, when the pests can occur on different plants.

Photoselective exclusion netting in apple orchards: effectiveness against pests and impact on beneficial arthropods, fungal diseases and fruit quality.

BACKGROUND: Frequent pesticide treatments in fruit orchards increase hazards for workers, consumers and the environment. Moreover, the indiscriminate and excessive use of pesticides often induces resistance in pests. In the past few years, physical exclusion strategies have been proposed as an alternative for the control of insect pests. The goal of this study was to evaluate the effectiveness of anti-hail photoselective netting in protecting apples against key and emerging pests, as well as the impact on beneficial arthropods, fungal diseases and fruit quality. RESULTS: In netted plots, a significant reduction in pest populations, i.e. fruit moths, Halyomorpha halys (Stål) and Drosophila suzukii (Matsumura), was recorded in comparison with un-netted controls. Moreover, the damage on fruits caused by H. halys was reduced up to 62% compared with insecticidal treatments. The net did not negatively affect the abundance of predators and the incidence of post-harvest rot. In addition, the incidence of bitter pit on apple was reduced up to 52%. Furthermore, fruit quality was unaffected by the net coverage (both at harvest and after 4 months of storage). CONCLUSION: Anti-hail photoselective pearl netting proved a promising exclusion system that can prevent attack by more than one insect pest at a time, allowing for a strong reduction in insecticide treatments and relative costs. At the same time, the netting did not negatively influence the presence of predators, the incidence of fungal disease or fruit quality. © 2019 Society of Chemical Industry.

Attraction of Halyomorpha halys (Hemiptera: Pentatomidae) haplotypes in North America and Europe to baited traps.

Halyomorpha halys is a global invasive species, native to Southeast Asia, that is threatening agriculture in invaded regions. Our objectives were to: 1) establish the attractiveness of semiochemical stimuli paired with field-deployed traps in Europe (Greece, Hungary, Italy, and Switzerland), compared with Maryland, USA, and 2) identify H. halys haplotypes recovered from traps at each location. We found qualitatively identical patterns of capture between sites located across Europe and in Maryland, USA. In both regions, captures of H. halys adults indicated a synergistic response to traps baited with the two component H. halys aggregation pheromone, and pheromone synergist, methyl (2E, 4E, 6Z)-decatrienoate when compared with either individually. Haplotype diversity in Europe based on trapped specimens was much greater than the USA, with five new haplotypes described here, probably indicating ongoing invasion and re-introduction of H. halys. By contrast, a single, previously identified haplotype was trapped in Maryland, USA, representing a single introduction. All H. halys haplotypes responded to each semiochemical in apparent proportion to their frequency in the overall population based on independently derived information from prior work. Taken together, these data suggest that pheromone-based technology will be of global utility for the monitoring of this important invasive species.

Efficient detection of Frankliniella schultzei (Thysanoptera, Thripidae) by cytochrome oxidase I gene (mtCOI) direct sequencing and real-time PCR

ABSTRACT Identification of Thysanoptera is based mainly on external morphology examination that can be time-consuming and difficult for non taxonomic experts. In this work, we propose a rapid and efficient molecular method to identify Frankliniella schultzei, an important and widespread pest thrips vector of tospoviruses in South America countries. Species-specific primers designed in the mitochondrial cytochrome oxidase I gene (mtCOI) were optimized for detection by conventional PCR and real-time PCR. The primers were tested on immature and adult thrips collected from crops and weeds found in São Paulo State. All samples collected were identified as F. schultzei, indicating the high prevalence of this species as vector of tospoviruses in Brazilian fields.

Adaptation of indigenous larval parasitoids to Tuta absoluta (Lepidoptera: Gelechiidae) in Italy.

Tuta absoluta (Meyrick) (Lepidoptera: Gelechiidae) is a serious threat to tomato (Solanum lycopersicum L.) crops in South America. In Europe, after its first detection in Spain in 2006, it rapidly spread through the Mediterranean basin, reaching Italy 2 yr later. The aim of our work was to find indigenous effective biological control agents and to evaluate their potential role in the control of larval populations of T. absoluta in controlled conditions. Nine species of larval parasitoids emerged from field-collected tomato leaves infested by T. absoluta. The most abundant, Necremnus near artynes (Walker) and Necremnus near tidius (Walker) (Hymenoptera: Eulophidae), were tested in laboratory parasitism trials. Furthermore, because the species N. artynes and N. tidius are each reported in literature as an ectoparasitoid of Cosmopterix pulchrimella Chambers (Lepidoptera: Cosmopterigidae) on upright pellitory plants, olfactometer bioassays were performed to assess the response of our parasitoids to the odors of tomato and pellitory leaves infested by T absoluta and C. pulchrimella, respectively, compared with healthy ones. Both Necremnus species showed good adaptation to the invasive pest, and we observed a high larval mortality of T. absoluta because of host feeding and parasitism. Even olfactory responses highlighted a preference of both wasps for tomato plants infested by the exotic pest. These preliminary results demonstrated a high suitability of these indigenous natural enemies for controlling T. absoluta. Further investigations are needed to confirm their role as potential biological agents in commercial tomato plantations.

Tospoviruses in the Mediterranean area.

Tospoviruses are among the most serious threats to vegetable crops in the Mediterranean basin. Tospovirus introduction, spread, and the diseases these viruses cause have been traced by epidemiological case studies. Recent research has centered on the close relationship between tospoviruses and their arthropod vectors (species of the Thripidae family). Here, we review several specific features of tospovirus-thrips associations in the Mediterranean. Since the introduction of Frankliniella occidentalis in Europe, Tomato spotted wilt virus (TSWV) has become one of the limiting factors for vegetable crops such as tomato, pepper, and lettuce. An increasing problem is the emergence of TSWV resistance-breaking strains that overcome the resistance genes in pepper and tomato. F. occidentalis is also a vector of Impatiens necrotic spot virus, which was first observed in the Mediterranean basin in the 1980s. Its importance as a cause of vegetable crop diseases is limited to occasional incidence in pepper and tomato fields. A recent introduction is Iris yellow spot virus, transmitted by the onion thrips Thrips tabaci, in onion and leek crops. Control measures in vegetable crops specific to Mediterranean conditions were examined in the context of their epidemiological features and tospovirus species which could pose a future potential risk for vegetable crops in the Mediterranean were discussed.

Integrated management of Thrips tabaci (Thysanoptera: Thripidae) on onion in north-western Italy: basic approaches for supervised control.

BACKGROUND: Thrips tabaci Lindeman (Thysanoptera: Thripidae) is a major pest on onion, Allium cepa L., worldwide. In 2010, research was conducted in a commercial onion field in north-western Italy in order (i) to evaluate the efficacy of different insecticides and of the SAR activator acibenzolar-S-methyl, (ii) to correlate thrips infestation levels with bulb size and weight at harvest and (iii) to implement a reliable thrips sampling method. Efficacy of the three active ingredients spinosad, lambda-cyhalothrin and acibenzolar-S-methyl on local thrips populations were also evaluated in laboratory bioassays. RESULTS: During field surveys, the highest and the lowest thrips infestations were observed in plots treated with lambda-cyhalothrin and with spinosad and acibenzolar-S-methyl respectively. The effectiveness of spinosad was also confirmed in laboratory bioassays. At harvest, bulb size and weight did not significantly differ between treatments. A high correlation with visual inspection made plant beating a suitable sampling method for routine practice, enabling a good estimate of thrips infestation. CONCLUSION: Damage caused by thrips is often not severe enough to warrant the frequent pesticide applications the crops receive in north-western Italy. The use of spinosad and acibenzolar-S-methyl is suggested as an alternative to conventional insecticides for the preservation of natural enemies.

Impact of Eurygaster maura (Heteroptera: Scutelleridae) feeding on quality of bread wheat in relation to attack period.

Sunn pest (or cereal bug) (Heteroptera: Pentatomidae and Scutelleridae) infestations of wheat, Triticum aestivum L., in the grain filling stage have the potential to adversely affect the quality of harvested grain for bread making. In the absence of resistant wheat cultivars, producers must rely on chemical control to protect their crop from sunn pest infestations. To implement an efficient environment friendly control strategy, there is a need to pinpoint the relationships between the timing of the bug attack and gluten degradation. Recent outbreaks of Eurygaster maura (L.) in northwestern Italy have increased the local concern toward this problem. A 3-yr study was carried out by caging plants of two bread wheat cultivars, characterized by different seed texture and bread-making quality, and introducing adults of E. maura in four periods corresponding to different grain filling stages: heading, early milk-ripe, milk-ripe, and late milk-ripe. The degree of bread-making quality depletion was assessed by analytical and biochemical methods and related to the attack period. Using analysis of variance, significant differences were found in the quality traits of kernels attacked by E. maura in different grain filling stages, the maximum damage occurring with bug feeding at the late milk-ripe stage. Biochemical investigations on gluten confirmed analytical results; in grain samples infested at the late milk-ripe stage, SDS gel electrophoresis revealed the degradation of some components of the high-molecular-weight glutenins, and high-performance liquid chromatography analyses showed a breakdown of the first peak of the insoluble fraction, mainly containing polymeric proteins highly related to dough strength.

Field Evaluation of Tomato Hybrids Engineered with Tomato spotted wilt virus Sequences for Virus Resistance, Agronomic Performance, and Pollen-Mediated Transgene Flow.

ABSTRACT Tomato hybrids obtained from homozygous progeny of line 30-4, engineered for Tomato spotted wilt virus (TSWV) resistance, were tested under field conditions in two locations with their corresponding nontransgenic hybrids. No transgenic hybrid became infected, but 33 to 50% of plants of each nontransgenic hybrid became infected with a severe reduction of marketable fruit production. The transgenic hybrids conformed to the standard agronomic characteristics of the corresponding nontransgenic ones. Fruit were collected from the nontransgenic plots included in the experimental field and from border rows, and seed were used to estimate the flow of the transgene via pollen. No transgene flow was detected in the protected crops; however, in the open field experiment, 0.32% of tomato seedlings were found to contain the genetic modification. Immunity to TSWV infection in 30-4 hybrids was confirmed in laboratory conditions using mechanical inoculation and grafting. Thrips inoculation in leaf discs of line 30-4 demonstrated that TSWV replication was inhibited at the primary infection site but not in leaf discs of a commercial hybrid containing the naturally occurring resistance gene Sw-5. Due to the high economic value of tomato crops worldwide and the importance of TSWV, the engineered resistance described here is of practical value for breeding into cultivars of commercial interest, because it could be combined with naturally occurring resistance, thus greatly reducing the ability of the virus to develop resistance-breaking strains.

Detection of Tomato spotted wilt virus in its vector Frankliniella occidentalis by reverse transcription-polymerase chain reaction.

A method for rapid and reliable detection of Tomato spotted wilt virus (TSWV) (Tospovirus, Bunyaviridae) in its vector Frankliniella occidentalis (Thysanoptera Thripidae) would be a useful tool for studying the epidemiology of this virus. A RT-PCR method developed for this purpose is reported. The method was tested on thrips involved in laboratory transmission trials and on thrips collected in the field, whose capability to transmit TSWV was checked previously by leaf disk assays. The RT-PCR results were consistent with the results obtained by the leaf disk assays. Among thrips involved in laboratory experiments, 97% of the adults that transmitted TSWV were positive by RT-PCR; as did some non-transmitter adults reacted, whereas among field-collected thrips only the individuals able to transmit were positive by RT-PCR. In addition, healthy thrips were allowed to feed as adults on virus-infected leaves for 48 h, and then examined by RT-PCR immediately or after starving or feeding on virus-free plants for various times, to determine if virus ingested (but not transmissible) was also detectable. The virus was detectable immediately after the feed or within 12 and 24 h for individuals starved or fed on virus-free plants, respectively, but not after those periods. Thus, the method could detect rapidly and reliably the virus in vectors from the field, providing 24 h of starving to avoid positive RT-PCR results from thrips simply carrying the virus.

Field Isolates of Tomato spotted wilt virus Overcoming Resistance in Pepper and Their Spread to Other Hosts in Italy.

Isolates of Tomato spotted wilt virus (TSWV) severely and systemically infecting commercial pepper cultivars with resistance introgressed from Capsicum chinense PI152225 were found in Albenga (northwestern Italy) in July 2000. Experimentally, these resistance-breaking (RB) isolates overcame the resistance in C. chinense PI152225, but they produced infection in other hosts similarly to non-RB isolates from the same area. The RB isolates were indistinguishable from TSWV by serology and electron microscopy, and they were efficiently transmitted by Frankliniella occidentalis. Such isolates were recovered on the same farm in tomato, pepper, and artichoke 2 and 12 months later, suggesting natural spread from the resistant plants and survival. The RB isolates survived in experimental mixed infections with a non-RB isolate in susceptible pepper and C. chinense, but cross-protection in pepper acted against them. Commercial TSWV-resistant pepper but not resistant tomato cultivars from different companies were susceptible to these RB isolates after mechanical inoculation. Similar isolates were not detected among TSWV samples collected from 1993 to 2000 in the area. The management of TSWV and thrips using resistant pepper cultivars is discussed.