Morphological characterization of the antenna of Torymus sinensis (Hymenoptera: Torymidae) and a comparison within the superfamily Chalcidoidea.

The parasitoid Torymus sinensis (Hymenoptera: Torymidae) has been successfully used in Italy since 2005 for biological control of the invasive cynipid Dryocosmus kuriphilus (Hymenoptera: Cynipidae), highly destructive for the economically relevant Castanea sativa (Fagales: Fagaceae). In order to investigate the morphological aspects related to sensorial behavior, a fine morphology study of the antennae and their sensilla was conducted by scanning electron microscopy on both sexes of T. sinensis. The antennae, composed of a scape, a pedicel and a flagellum with ten flagellomeres, had chaetic sensilla of six subtypes, placoid sensilla of three subtypes, trichoid sensilla, sensilla with a roundish grooved tip, and coeloconic sensilla. The chaetic sensilla of the first three subtypes were found in the scape and in the pedicel, and those of the last three subtypes, together with trichoid, roundish grooved tip and coeloconic sensilla, were found only on flagellomeres. Sexual dimorphism was detected in the morphology of the proper pedicel and the flagellum, and in the presence and distribution of the sensilla and their subtypes. The morphological aspects of the antenna of T. sinensis and of its sensilla were compared with those found in the family Torymidae and in other families of the extremely diverse superfamily Chalcidoidea.

Description of Calling, Courtship and Mating Behaviour of Six Species of the Genus Zyginidia (Auchenorrhyncha: Cicadellidae).

The vibrational communication and mating behaviour of the graminicolous leafhoppers Zyginidia pullula, Z. ribauti, Z. scutellaris, Z. serpentina, Z. sohrab, and Z. biroi were investigated to explain why the first five species hybridize. Z. biroi was used as a control species. All species behaved in the same way and no significant statistical differences were detected with regard to male calls, while female calls and the male courtship song differed in Z. biroi, thus showing that a specific pre-mating isolation mechanism was used by the latter species and the first five ones lacked such a mechanism. In addition, Z. sohrab is missing in Italy, while the other species live allopatrically in Italy, with the only exceptions being Z. serpentina and Z. biroi, which live in Sicily and are often found in sympatry, and Z. scutellaris and Z. biroi, which live in Sardinia. All these species can be distinguished by means of male genital appendages; however, Z. biroi is longer and has a different body colour. The existence of natural hybrids of Z. pullula, Z. ribauti, and Z. scutellaris in the Italian peninsula and their hybridization in the laboratory with Z. serpentina and Z. sohrab require the investigation of possible post-mating reproductive barriers before re-considering their systematic validity.

First record of the non-native Osbornellusauronitens (Provancher, 1889) (Hemiptera, Cicadellidae, Deltocephalinae) in Italy.

Background: Globalisation and international trade, in particular, are the major drivers of introduction and the spread of non-native species. To date, more than 30 species of non-native Hemiptera Auchenorrhyncha have been accidentally introduced into Europe. Some species are invasive with important repercussions primarily for agricultural activities, while almost no information exists on their impacts within natural ecosystems. Therefore, early detection of non-native species and their subsequent monitoring are extremely important actions to undertake. New information: The North American Osbornellusauronitens (Provancher, 1889), firstly recorded for the Palearctic and Europe in Switzerland in 2016, is recorded in Italy for the first time on the basis of 77 specimens collected between August 2015 and October 2022.

Popillia japonica - Italian outbreak management.

Popillia japonica, a priority pest for the EU, was first detected in Northern Italy in 2014. Since its discovery, the outbreak extended over an area of more than 16,000 square kilometers in Northern Italy and Southern Switzerland. In this review, we summarize the state-of-the-art of research conducted in Italy on both the spreading capacity and control measures of P. japonica. Chemical, physical, and biological control measures deployed since its detection are presented, by highlighting their strengths and weaknesses. An in-depth study of the ecosystems invaded by P. japonica disclosed the presence and pathogenicity of natural strains of entomopathogenic fungi and nematodes, some of which have shown to be particularly aggressive towards the larvae of this pest under laboratory conditions. The Plant Health authorities of the Lombardy and Piedmont regions, with the support of several research institutions, played a crucial role in the initial eradication attempt and subsequently in containing the spread of P. japonica. Control measures were performed in the infested area to suppress adult populations of P. japonica by installing several traps (e.g., for mass trapping, for auto-dissemination of the fungus Metarhizium anisopliae, and "attract & kill"). For larval control, the infested fields were treated with commercial strains of the entomopathogenic fungus M. anisopliae and nematode Heterorhabditis bacteriophora. Future studies will aim at integrating phenological and spread models developed with the most effective control measures, within an ecologically sustainable approach.

Assessment of Chestnut Gall Toughness: Implications for a Biocontrol Agent.

(1) Torymus sinensis, the biocontrol agent of the Asian chestnut gall wasp Dryocosmus kuriphilus, is univoltine, but in NW Italy a small percentage of individuals exhibits a prolonged diapause, mainly as late instar larva. (2) In 2020, the diapause was investigated to evaluate its trend over the years. Due to the low survival rate of diapausing T. sinensis adults, the seasonal variation in the galls' toughness was evaluated, thus assuming that dry galls over time can negatively affect emergence. The window of vulnerability of the gall wasp galls was also evaluated in controlled conditions. (3) The results showed that the average number of second year T. sinensis emerging per 100 cells was 0.41 ± 0.05, and dead adults accounted for 4.1 ± 0.23 per 100 cells. Gall toughness resulted in lower values for galls collected in May and June, and then gradually increased over time. In general, no difference was detected in the wall toughness of galls formed during the previous year when compared to current-year dry galls. Oviposition was recorded on all the tested galls collected in May and June, and no difference in the number of oviposition events was detected. Conversely, no oviposition was observed in July. Comparing the number of oviposition events by T. sinensis and the gall toughness, a negative correlation was found (R2 = -0.99). (4) The present findings contribute descriptive information on this gall's structural traits, and the influence on gall wasp management is also discussed.

Pheromone-Mediated Mating Disruption as Management Option for Cydia spp. in Chestnut Orchard.

(1) Background: Pheromone-based devices are successfully used to control insect pests in agriculture. (2) Methods: Investigations were conducted to evaluate the effectiveness of mating disruption (MD) to control the chestnut tortrix moths, Cydia fagiglandana and Cydia splendana. Surveys were performed in northern Italy in 2019-2020. MD was carried out using the pheromone dispenser Ecodian® CT. The effectiveness of MD was assessed by recording male adult catches in pheromone-baited sticky traps and larvae in chestnut fruits, comparing MD and control plots. (3) Results: The total number of trapped males was significantly lower in MD plots than in control ones, for all sites and years. Trap catch suppression in MD plots averaged 89.5% and 93.8% for C. fagiglandana and 57.4% and 81% for C. splendana in 2019 and 2020, respectively. The larval infestation rate in fruits did not vary between plots except for one site where a reduction of about 71% in the MD plot was recorded in 2019. (4) Conclusions: Low catches in MD plots turned out to be a good measure of the effectiveness of communication disruption, but no satisfactory data have been obtained regarding fruit infestation, highlighting how the reduction of male catches cannot always be considered as a reliable indicator of successful control. Specific investigations about background population density, dispersal and mating/oviposition behavior are thus essential for a viable management strategy.

Compartmentalization of bacterial and fungal microbiomes in the gut of adult honeybees.

The core gut microbiome of adult honeybee comprises a set of recurring bacterial phylotypes, accompanied by lineage-specific, variable, and less abundant environmental bacterial phylotypes. Several mutual interactions and functional services to the host, including the support provided for growth, hormonal signaling, and behavior, are attributed to the core and lineage-specific taxa. By contrast, the diversity and distribution of the minor environmental phylotypes and fungal members in the gut remain overlooked. In the present study, we hypothesized that the microbial components of forager honeybees (i.e., core bacteria, minor environmental phylotypes, and fungal members) are compartmentalized along the gut portions. The diversity and distribution of such three microbial components were investigated in the context of the physico-chemical conditions of different gut compartments. We observed that changes in the distribution and abundance of microbial components in the gut are consistently compartment-specific for all the three microbial components, indicating that the ecological and physiological interactions among the host and microbiome vary with changing physico-chemical and metabolic conditions of the gut.

An integrated transcriptomic and proteomic approach to identify the main Torymus sinensis venom components.

During oviposition, ectoparasitoid wasps not only inject their eggs but also a complex mixture of proteins and peptides (venom) in order to regulate the host physiology to benefit their progeny. Although several endoparasitoid venom proteins have been identified, little is known about the components of ectoparasitoid venom. To characterize the protein composition of Torymus sinensis Kamijo (Hymenoptera: Torymidae) venom, we used an integrated transcriptomic and proteomic approach and identified 143 venom proteins. Moreover, focusing on venom gland transcriptome, we selected additional 52 transcripts encoding putative venom proteins. As in other parasitoid venoms, hydrolases, including proteases, phosphatases, esterases, and nucleases, constitute the most abundant families in T. sinensis venom, followed by protease inhibitors. These proteins are potentially involved in the complex parasitic syndrome, with different effects on the immune system, physiological processes and development of the host, and contribute to provide nutrients to the parasitoid progeny. Although additional in vivo studies are needed, initial findings offer important information about venom factors and their putative host effects, which are essential to ensure the success of parasitism.

Models Applied to Grapevine Pests: A Review.

This paper reviews the existing predictive models concerning insects and mites harmful to grapevine. A brief conceptual description is given on the definition of a model and about different types of models: deterministic vs. stochastics, continuous vs. discrete, analytical vs. computer-based, and descriptive vs. data-driven. The main biological aspects of grapevine pests covered by different types of models are phenology, population growth and dynamics, species distribution, and invasion risk. A particular emphasis is put on forecasting epidemics of plant disease agents transmitted by insects with sucking-piercing mouthparts. The most investigated species or groups are the glassy-winged sharpshooter Homalodisca vitripennis (Germar) and other vectors of Xylella fastidiosa subsp. fastidiosa, a bacterium agent of Pierce's disease; the European grape berry moth, Lobesia botrana (Denis and Schiffermuller); and the leafhopper Scaphoideus titanus Ball, the main vector of phytoplasmas agents of Flavescence dorée. Finally, the present and future of decision-support systems (DSS) in viticulture is discussed.

Disruption of Host-Symbiont Associations for the Symbiotic Control and Management of Pentatomid Agricultural Pests-A Review.

The family Pentatomidae (Hemiptera: Heteroptera) includes several invasive stink bug species capable to attack a large number of wild and cultivated plants, causing several damages to different crops. Pentatomids rely on obligate symbiotic associations with bacteria of the family Enterobacteriaceae, mainly of the genus Pantoea. A distinctive trait of these associations is the transmission route: during oviposition, females smear egg masses with symbiont-containing secretions, which are ingested by newly hatched nymphs, allowing the symbiont to pass through their digestive tract and establish in the crypts of the posterior midgut. Preventing newborns from orally acquiring symbionts seriously affects their fitness and survival. This symbiont inheritance process can be manipulated to develop innovative pest control measures by sterilization of egg masses prior to nymph hatching. This review summarizes the recent knowledge advances concerning the gut primary symbionts of pentatomids, with a specific focus on the most troubling pest species for agriculture. Current understanding of host colonization dynamics in pentatomids is presented, as well as the phenotypic effects determined in different insect species by the alteration of vertical transmission. Details on the current knowledge on the whole bacterial communities accompanying primary symbionts are analyzed. The recent research exploiting the perturbation of symbiont acquisition by pentatomid nymphs is discussed, by considering published work on laboratory and field trials with several active substances. These translational strategies are presently regarded as promising for limiting the populations of many important pentatomid pests in a sustainable way.

Performance of Pheromone-Baited Traps to Monitor the Seasonal Abundance of Tortrix Moths in Chestnut Groves.

(1) Background: Pammene fasciana (L.), Cydia fagiglandana (Zeller), and C. splendana (Hübner) (Lepidoptera: Tortricidae) are considered key moth pests of chestnut in Europe. (2) Methods: Investigations were performed in 2018-2019 in northern Italy. Sticky traps and commercially available pheromones were used for monitoring; moreover, two experimental pheromone blends were tested. All specimens were identified according to male genitalia and molecular analyses. Newly formed chestnut husks and fruits were randomly collected to evaluate the presence of larvae and/or feeding damage, by comparing it to trap catches. (3) Results: P. fasciana was present in all the sites, whereas Cydia species were recorded in three sites of six, with differences in abundance related to pheromone blends studied. Several non-target species, such as Oegoconia novimundi (Busck) and Cydia ilipulana (Walsingham), were present. Data about the seasonal flight activity are provided. (4) Conclusions: This research contributes to ascertaining the presence and abundance of tortrix moths in Italian chestnut groves, and the presence of non-target species highlights the risk of overestimating catches. Fruit damage recorded did not always reflect catches made by pheromone traps, suggesting that monitoring may underestimate the real size of moths' populations. All of the data acquired are important for planning specific control measures.

Update and New Epidemiological Aspects about Grapevine Yellows in Chile.

To date, phytoplasmas belonging to six ribosomal subgroups have been detected to infect grapevines in Chile in 36 percent of the sampled plants. A new survey on the presence of grapevine yellows was carried out from 2016 to 2020, and 330 grapevine plants from the most important wine regions of the country were sampled and analyzed by nested PCR/RFLP analyses. Phytoplasmas enclosed in subgroups 16SrIII-J and 16SrVII-A were identified with infection rates of 17% and 2%, respectively. The vineyards in which the phytoplasma-infected plants were detected were further inspected to identify alternative host plants and insects of potential epidemiological relevance. Five previously unreported plant species resulted positive for 16SrIII-J phytoplasma (Rosa spp., Brassica rapa, Erodium spp., Malva spp. and Rubus ulmifolius) and five insect species were fully or partially identified (Amplicephalus ornatus, A. pallidus, A. curtulus, Bergallia sp., Exitianus obscurinervis) as potential vectors of 16SrIII-J phytoplasmas. The 16SrVII-A phytoplasmas were not detected in non-grape plant species nor in insects. This work establishes updated guidelines for the study, management, and prevention of grapevine yellows in Chile, and in other grapevine growing regions of South America.

Palaearctic Egg Parasitoids Interaction to Three Grapevine Exotic Pests in Northwestern Italy: A New Association Involving Metcalfa pruinosa.

The most important exotic leafhopper pests currently affecting the Italian vineyards are the leafhoppers Scaphoideus titanus, Orientus ishidae and the planthopper Metcalfa pruinosa. Their highest population density is detected in the uncultivated areas with wild grapevines. Should these habitats be considered only a problem or a potential resource for Palearctic entomophagy of these three exotic pests? The aim of this work was to study the biotopes and biocoenosis present in the Piedmontese vineyard agroecosystem, evaluating the parasitization rate and other crucial aspects for a possible application in biological control. Several specimens of egg-parasitoid wasps were obtained from filed-collected two-year-old grapevine canes. The most prevalent one belonged to the Oligosita collina group (Trichogrammatidae) emerged only from M. pruinosa eggs with a parasitization rate of over 40%. The new association is the first report of such a high level of parasitization on the flatid planthopper. The parasitization rate mainly relied on the host egg density and the abundance of plants suitable for the oviposition. A second parasitoid generation on the overwintering eggs is discussed, as well as other hypothesis. Furthermore, the parasitization rate was higher than the one showed by the dryinid Neodryinus typhlocybae, the control agent introduced in Italy under the biological control strategy, highlighting a possible implication in this biocoenosis. We assume that the egg parasitoid adaptation may contribute to M. pruinosa control.

Ability of Euscelidius variegatus to Transmit Flavescence Dorée Phytoplasma with a Short Latency Period.

Phytoplasma transmission takes place by insect vectors through an Acquisition Access Period (AAP), Latency Period (LP) and Inoculation Access Period (IAP). Generally, phytoplasmas are believed to be transmitted more efficiently by nymphs because they need a long LP to reach the salivary glands before becoming infective. The transmission can start from adults as well, but in this case a long LP may exceed the insect's lifespan. However, previous evidence has indicated that adults can undergo a shorter LP, even though little knowledge is available regarding the phytoplasma temporal dynamics during this period. Here, we investigate the minimum time required by the phytoplasma to colonize the vector midgut and salivary glands, and finally to be inoculated into a plant. We used the leafhopper Euscelidius variegatus to investigate the life cycle of flavescence dorée phytoplasma (FDP). Phytoplasma-free E. variegatus adults were left on broad beans (BBs) infected with FDP for an AAP of 7 days. Subsequently, they were individually transferred onto a healthy BB for seven different IAPs, each one lasting 24 h from day 8 to 14. Molecular analyses and fluorescence in situ hybridization were performed for FDP detection. FDP was found in the leafhopper midgut from IAP 1 with an infection rate reaching 50%, whereas in the salivary glands it was found from IAP 2 with an infection rate reaching 30%. FDP was also detected in BBs from IAP 4, with infection rates reaching 10%. Our results represent an important step to further deepen the knowledge of phytoplasma transmission and its epidemiology.

Development of High-Density Genetic Linkage Maps and Identification of Loci for Chestnut Gall Wasp Resistance in Castanea spp.

Castanea sativa is an important multipurpose species in Europe for nut and timber production as well as for its role in the landscape and in the forest ecosystem. This species has low tolerance to chestnut gall wasp (Dryocosmus kuriphilus Yasumatsu), which is a pest that was accidentally introduced into Europe in early 2000 and devastated forest and orchard trees. Resistance to the gall wasp was found in the hybrid cultivar 'Bouche de Bétizac' (C. sativa × C. crenata) and studied by developing genetic linkage maps using a population derived from a cross between 'Bouche de Bétizac' and the susceptible cultivar 'Madonna' (C. sativa). The high-density genetic maps were constructed using double-digest restriction site-associated DNA-seq and simple sequence repeat markers. The map of 'Bouche de Bétizac' consisted of 1459 loci and spanned 809.6 cM; the map of 'Madonna' consisted of 1089 loci and spanned 753.3 cM. In both maps, 12 linkage groups were identified. A single major QTL was recognized on the 'Bouche de Bétizac' map, explaining up to 67-69% of the phenotypic variance of the resistance trait (Rdk1). The Rdk1 quantitative trait loci (QTL) region included 11 scaffolds and two candidate genes putatively involved in the resistance response were identified. This study will contribute to C. sativa breeding programs and to the study of Rdk1 genes.

Developmental stages and gut microenvironments influence gut microbiota dynamics in the invasive beetle Popillia japonica Newman (Coleoptera: Scarabaeidae).

Popillia japonica Newman (Coleoptera: Scarabaeidae) is a highly polyphagous invasive beetle originating from Japan. This insect is highly resilient and able to rapidly adapt to new vegetation. Insect-associated microorganisms can play important roles in insect physiology, helping their hosts to adapt to changing conditions and potentially contributing to an insect's invasive potential. Such symbiotic bacteria can be part of a core microbiota that is stably transmitted throughout the host's life cycle or selectively recruited from the environment at each developmental stage. The aim of this study was to investigate the origin, stability and turnover of the bacterial communities associated with an invasive population of P. japonica from Italy. Our results demonstrate that soil microbes represent an important source of gut bacteria for P. japonica larvae, but as the insect develops, its gut microbiota richness and diversity decreased substantially, paralleled by changes in community composition. Notably, only 16.75% of the soil bacteria present in larvae are maintained until the adult stage. We further identified the micro-environments of different gut sections as an important factor shaping microbiota composition in this species, likely due to differences in pH, oxygen availability and redox potential. In addition, P. japonica also harboured a stable bacterial community across all developmental stages, consisting of taxa well known for the degradation of plant material, namely the families Ruminococcacae, Christensenellaceae and Lachnospiraceae. Interestingly, the family Christensenallaceae had so far been observed exclusively in humans. However, the Christensenellaceae operational taxonomic units found in P. japonica belong to different taxonomic clades within this family.

Activation of Immune Genes in Leafhoppers by Phytoplasmas and Symbiotic Bacteria.

Insect immunity is a crucial process in interactions between host and microorganisms and the presence of pathogenic, commensal, or beneficial bacteria may result in different immune responses. In Hemiptera vectors of phytoplasmas, infected insects are amenable to carrying high loads of phytopathogens, besides hosting other bacterial affiliates, which have evolved different strategies to be retained; adaptation to host response and immunomodulation are key aspects of insect-symbiont interactions. Most of the analyses published to date has investigated insect immune response to pathogens, whereas few studies have focused on the role of host immunity in microbiota homeostasis and vectorial capacity. Here the expression of immune genes in the leafhopper vector of phytoplasmas Euscelidius variegatus was investigated following exposure to Asaia symbiotic bacteria, previously demonstrated to affect phytoplasma acquisition by leafhoppers. The expression of four genes related to major components of immunity was measured, i.e., defensin, phenoloxidase, kazal type 1 serine protease inhibitor and Raf, a component of the Ras/Raf pathway. The response was separately tested in whole insects, midguts and cultured hemocytes. Healthy individuals were assessed along with specimens undergoing early- and late-stage phytoplasma infection. In addition, the adhesion grade of Asaia strains was examined to assess whether symbionts could establish a physical barrier against phytoplasma colonization. Our results revealed a specific activation of Raf in midguts after double infection by Asaia and flavescence dorée phytoplasma. Increased expression was observed already in early stages of phytoplasma colonization. Gut-specific localization and timing of Raf activation are consistent with the role played by Asaia in limiting phytoplasma acquisition by E. variegatus, supporting the involvement of this gene in the anti-pathogen activity. However, limited attachment capability was found for Asaia under in vitro experimental conditions, suggesting a minor contribution of physical phytoplasma exclusion from the vector gut wall. By providing evidence of immune modulation played by Asaia, these results contribute to elucidating the molecular mechanisms regulating interference with phytoplasma infection in E. variegatus. The involvement of Raf suggests that in the presence of reduced immunity (reported in Hemipterans), immune genes can be differently regulated and recruited to play additional functions, generally played by genes lost by hemipterans.

Development, Spatial Distribution, and Presence on Grapevine of Nymphs of Orientus ishidae (Hemiptera: Cicadellidae), a New Vector of Flavescence Dorée Phytoplasmas.

Orientus ishidae (Matsumura) (subfamily Deltocephalinae) is an Asian species now widespread in Europe, and a vector of 16SrV phytoplasmas agents of grapevine Flavescence dorée (FDP). Embryonic and post-embryonic development, spatial distribution, and relationships with grapevine of nymphs were studied under field and laboratory conditions. Egg-hatching dynamics and post-embryonic development of nymphs were studied by collecting grapevine wood from managed and unmanaged vineyards (including bot European Vitis vinifera L., and wild American rootstocks) and storing it inside rearing cages at T = 21-23°C. Field sampling of nymphs were made on both grapevine and two elective host plants of O. ishidae: hazelnut and hornbeam. Taylor's Power Law was applied to assess the aggregation coefficient of early- (first and second) and late- (third to fifth) life instars on leaves and shoots of host plants. More nymphs were obtained from wood collected in unmanaged rather than managed vineyards. Under lab conditions, the embryonic development lasted 34-48 d, whereas the whole post-embryonic development averaged 27 d. Under field conditions, early instars peaked at the end of May, and late instars peaked 2-4 wk later. The aggregation patterns decreased from early to late instars, and from leaves to shoots. Very few nymphs were observed on unmanaged grapevine, either European or American, and none on managed European grapevine. Some behavioral and FDP epidemiological consequences of the results obtained are discussed.

Bacterial diversity shift determined by different diets in the gut of the spotted wing fly Drosophila suzukii is primarily reflected on acetic acid bacteria.

The pivotal role of diet in shaping gut microbiota has been evaluated in different animal models, including insects. Drosophila flies harbour an inconstant microbiota among which acetic acid bacteria (AAB) are important components. Here, we investigated the bacterial and AAB components of the invasive pest Drosophila suzukii microbiota, by studying the same insect population separately grown on fruit-based or non-fruit artificial diet. AAB were highly prevalent in the gut under both diets (90 and 92% infection rates with fruits and artificial diet respectively). Fluorescent in situ hybridization and recolonization experiments with green fluorescent protein (Gfp)-labelled strains showed AAB capability to massively colonize insect gut. High-throughput sequencing on 16S rRNA gene indicated that the bacterial microbiota of guts fed with the two diets clustered separately. By excluding AAB-related OTUs from the analysis, insect bacterial communities did not cluster separately according to the diet, suggesting that diet-based diversification of the community is primarily reflected on the AAB component of the community. Diet influenced also AAB alpha-diversity, with separate OTU distributions based on diets. High prevalence, localization and massive recolonization, together with AAB clustering behaviour in relation to diet, suggest an AAB role in the D. suzukii gut response to diet modification.

The genus Cixius Latreille, 1804 (Hemiptera, Fulgoromorpha, Cixiidae) in Lebanon with the description of two new species.

A first list of Cixius fauna of Lebanon is here provided with the description of two new species: one within the subgenus Acanthocixius, Cixius bifidispinus sp. nov. and one within the subgenus Ceratocixius Cixius superremotus sp. nov. The standard morphological description is supplemented by sequence data of the mitochondrial cytochrome c oxidase subunit I (COI) gene fragment. Some notes on their distribution and economic importance are also reported.