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Olive (Olea europaea L.) is a traditional crop of great socioeconomic importance in the Mediterranean Basin. In 2021, a survey was performed in an Italian nursery located in Central Italy where unusual symptoms and widespread dieback had been observed on young potted olive trees. Twigs and branches of all plants affected by dryness showed the presence of the alien bark beetle Xylosandrus compactus. Woody tissues colonized by X. compactus were transferred into the laboratory and several fungi such as Cladosporium perangustum and Neofusicoccum parvum were then identified. In addition to these species, Geosmithia pallida was also reported for the first time in Italy on this plant. Studies on this bark beetle and its associated fungi are currently in progress to further evaluate potential risks for olive cultivation. Key Words: Olea europea, fungi, bark beetles.

Bark and wood-boring beetles feeding on coniferous and deciduous trees in different forest ecosystems are often associated with various species of fungi. In 2019, widespread attacks of Liparthrum colchicum Semenov (Coleoptera Curculionidae Scolytinae) were observed on Laurus nobilis L. in Tuscany (Italy). Samples of colonized terminal twigs were collected to investigate the presence of phytopathogenic fungi associated with the scolytid. Two different colonies of the Geosmithia genus were identified as Geosmithia pallida and Geosmithia langdonii. To our knowledge this is the first report of G.pallida and G. langdonii associated with L. colchicum on Bay tree, in Italy.

Liparthrum colchicumSemenov (Coleoptera Curculionidae Scolytinae) is a relatively unknown monophagous insect species primarily associated with thin dried twigs (about 0.5-1cm in diameter) of Laurel (Laurus nobilis). After being caught in traps located at Italian ports, it was initially considered an intercepted species alien to the country. However, recent findings, suggest that this beetle may have a different origin. Several records from different sites of Tuscany and Latium showed that it could be relatively widespread in Italy. These observations strengthened the hypothesis that this scolytid, harmless to L. nobilis, might be already established or even native to the country. Therefore, we argue that it could have been easily overlooked thus far, as it thrives only in twigs already wilted due to different environmental factors. Moreover, field observations, carried out in the two Italian Regions, evidenced high frequencies of its infestations in Laurel twigs previously colonized by Xylosandrus compactus. Notes on the taxonomy and biology of this species are also reported in the present wo

Olive cultivation is of great economic, ecological, and cultural interest in Italy, as well as in the rest of the Mediterranean basin. Among the pests of olive trees, several groups of insects, mites, and nematodes have been reported. Phytoparasitic nematodes especially of the genera Meloidogyne, Pratylenchus, Helicotylenchus, Xiphinema, Tylenchulus, Rotylenchulus, and Heterodera have usually been extracted from roots and soil around trees. On the other hand, no information is available concerning nematodes directly associated with the wood. At the end of September 2018, in a high-density cultivated olive grove in Tuscany (central Italy), several olive trees with decline symptoms were observed. Three Bursaphelenchus species, B. fungivorus, B. minutus, and B. sexdentati were extracted from the wood of one dead tree. Even though these species had already been reported in Italy, these findings were the first ones recorded in olive wood. Moreover, another undescribed Bursaphelenchusspecies was found associated with the bark beetle Hylesinus fraxini collected from olive trunks and branches. Further research is needed to investigate the role of insects and Bursaphelenchus spp. in the decline processes of olive trees.

Molecular assays based on qPCR TaqMan Probes were developed to identify three species of the genus Xylosandrus, X. compactus, X. crassiusculus and X. germanus (Coleoptera Curculionidae Scolytinae). These ambrosia beetles are xylophagous species alien to Europe, causing damages to many ornamental and fruiting trees as well as shrubs. DNA extraction was carried out from adults, larvae and biological samples derived from insect damages on infested plants. For X. compactus, segments of galleries in thin infested twigs were cut and processed; in the case of X. crassiusculus, raw frass extruded from exit holes was used, while DNA of X. germanus was extracted from small wood chips removed around insect exit holes. The assays were inclusive for the target species and exclusive for all the non-target species tested. The LoD was 3.2 pg/µL for the frass of X. crassiusculus and 0.016 ng/µL for the woody matrices of the other two species. Both repeatability and reproducibility were estimated on adults and woody samples, showing very low values ranging between 0.00 and 4.11. Thus, the proposed diagnostic assays resulted to be very efficient also on the woody matrices used for DNA extraction, demonstrating the applicability of the protocol in the absence of dead specimens or living stages.

Three molecular protocols using qPCR TaqMan probe, SYBR Green, and loop-mediated isothermal amplification (LAMP) methods were set up for the identification of larvae and adults of an African invasive moth, Thaumatotibia leucotreta (Meyrick, 1913) (Lepidoptera: Tortricidae). The DNA extracts from larval and adult samples of T. leucotreta were perfectly amplified with an average Ct value of 19.47 ± 2.63. All assays were demonstrated to be inclusive for T. leucotreta and exclusive for the nontarget species tested; the absence of false positives for nontarget species showed a 100% of diagnostic specificity and diagnostic sensitivity for all assays. With the SYBR Green protocol, the Cq values were only considered for values less than 22 (cutoff value) to prevent false-positive results caused by the late amplification of nonspecific amplicons. The limit of detection (LoD) for the qPCR probe protocol was equal to 0.02 pg/µl while a value equal to 0.128 pg/µl for the qPCR SYBR Green assay and LAMP method were established, respectively. The intrarun variabilities of reproducibility and repeatability in all the assays evaluated as CV%, ranged between 0.21 and 6.14, and between 0.33 and 9.52, respectively; the LAMP values were slightly higher than other assays, indicating a very low interrun variability. In order for an operator to choose the most desirable method, several parameters were considered and discussed. For future development of these assays, it is possible to hypothesize the setup of a diagnostic kit including all the three methods combined, to empower the test reliability and robustness.

A molecular diagnostic method using TaqMan probe qPCR is presented for the identification of Anoplophora chinensis (Förster) (Coleoptera: Cerambycidae) from whole body insects (adults and larvae) and frass samples stored under different conditions. The results showed a perfect amplification of DNA from all samples; the repeatability and reproducibility of the protocol were very good, with standard deviations of inter-run and intra-run variability less than or equal to 0.5. The assay allowed to discern all A. chinensis samples from those of the other non-target wood-borer species, with 100% correspondence to the homologous sequences. No amplification or cross reactions were observed with A. glabripennis (Motschulsky) (Coleoptera: Cerambycidae), which is the most related species among those tested. The protocol was validated by an internal blind panel test which showed a good correspondence between the results obtained by different operators in the same lab. The analytical sensitivity for the lab frass with the Probe qPCR, namely the lowest amount of A. chinensis DNA that can be detected (LoD), was 0.64 pg/µl with a Cq of 34.87. The use of indirect evidence for the identification of a pest is an important feature of the method, which could be crucial to detect the presence of wood-boring insects. This diagnostic tool can help prevent the introduction of A. chinensis into new environments or delimit existing outbreak areas thanks to indirect frass diagnosis.

Anoplophora glabripennis (Motschulsky, 1853), native to eastern Asia, is a destructive woodborer of many ornamental species, leading to the decline and the death of the attacked trees. In outbreak areas as Europe or North America, this pest is usually identified using morphological or molecular analyses of adult or larval specimens. However, the procedures for collecting A. glabripennis specimens from infested plants are too expensive and time consuming for routine screening. A noninvasive diagnostic tool based on frass discrimination is therefore crucial for the rapid identification of A. glabripennis at different development stages in the host. This article describes a rapid diagnostic protocol based on loop-mediated isothermal amplification (LAMP). DNA extracted from A. glabripennis frass was amplified with both visual and real-time LAMP and compared with those of nontarget species. The results show that the method is reliable and accurate and therefore could be a promising diagnostic tool in phytosanitary surveys.

In sweet chestnut cultivation, important ecosystem in Europe, the moderate sheep grazing practice may have some beneficial effects in organic farming. Its impact, on soil fertility, soil nematode community and level of fruit infested by tortricids and curculionids was evaluated in a case of study in Tuscany (Central Italy). An organic farm with sheep grazing system (SG) based on long-term field application was compared with an adjacent organic farm with poultry manure system (PM). In both managements, no treatment to control pests and diseases was applied and 0.19 kg /plant of nitrogen were distributed. SG increased twice the amount of organic C, total N and available P than PM. Moreover, the salinity effect of uric acid significantly increased the pH values in SG (from 5.0 to 6.2). The greatest accumulation of organic matter under SG provided stability in nematode community structure and the plant parasitic nematodes decreased. The analysis on the nematodes emerged from Galleria mellonella larvae, revealed that the abundance of native entomopathogenic nematodes was low and not effective to control insect pests. The sheep demonstrated to be the effective predators of tortricid and weevil larvae and in November, the total infestation was significantly lower in SG (34.3%) than PM (54.3%).

Aclees sp. cf. foveatus (Coleoptera: Curculionidae) is spreading in Central Italy, causing severe infestation on fig trees. There are very few information for this pest and no natural enemy is reported. Here, we report the first recovery of a natural enemy associated with this invasive weevil, a strain of the entomopathogenic fungus Beauveria bassiana . The potential use of entomopathogenic fungi and nematodes as biocontrol agents was tested against adults in laboratory trials. In agree with the detection in nature, only treatments with B. bassiana were able to control the insects. This result opens new frontiers for the environmental friendly control strategies against this weevil.

The Velvet Longhorned Beetle Trichoferus campestris (Faldermann, 1835) was intercepted for the first time in Italy, in the Naples harbour, during monitoring activities at entry points carried out in the context of the national project ASPROPI. T. campestris is widely polyphagous and able to colonize several woody species in both agricultural and forest environments. Moreover timber and solid wood items can be equally attractive to this pest. This additional report, in an area where other alien insect species had been identified in the past, emphasises how the national monitoring network represents a key element of the alert system designed to quickly counter the accidental introduction and spreading of exotic insect pests.

A new phoretic association between Bursaphelenchus minutus and the bark beetle Orthotomicus erosus collected on Pinus pinea is described for the first time. Nematode identification was based on morphological observation and biometric measurements of males and females. Moreover, B. minutus was characterized by ITS sequences and RFLP profile.

The attractiveness of different combinations of volatile compounds to several scolytid and cerambycid species, potential vectors of phytoparasitic nematodes of the genus Bursaphelenchus Fuchs, was investigated in pine woods of central Italy. The blend composed of α-pinene, ethanol, ipsenol, ipsdienol and 2-methyl-3-buten-2-ol was very efficient in trapping the cerambycids Monochamus galloprovincialis, Acanthocinus griseus and Arhopalus syriacus and the scolytids Ips sexdentatus, Orthotomicus erosus and Hylurgus ligniperda. This blend is proposed for operational surveys of Bursaphelenchus spp. associated with these beetles. Comparison of the capture levels with cross-vane and funnel traps baited with α-pinene, ethanol, ipsenol, ipsdienol, 2-methyl-3-buten-2-ol and cis-verbenol revealed the higher efficacy of the cross-vane traps for the above-mentioned cerambycids and the scolytid I. sexdentatus but greater effectiveness of the funnel traps in capturing H. ligniperda and O. erosus. Cross-vane traps were more efficient in capturing large numbers of these insects. Data on non-target xylophagous beetles and predator species are also reported.

Ricania speculum (Walker, 1851) (Hemiptera: Fulgoromorpha: Ricaniidae) is reported for the first time in Europe. Both nymphs and adults were observed from 2009 in several municipalities of Liguria (Italy). Since the species is extremely polyphagous and is a real pest for several crops in tropical and subtropical areas, the presence of this alien insect is noteworthy, representing a new possible threat for native species and human activities.

The male and the oviparous female of Cinara palaestinensis Hille Ris Lambers, the Aleppo pine aphid, are recorded for the first time, in populations on Pinus halepensis in Italy. Description and illustrations are provided together with additional notes on taxonomy, ecology and distribution of the species.

Ricania speculum (Walker, 1851) (Hemiptera: Fulgoromorpha: Ricaniidae) is reported for the first time in Europe. Both nymphs and adults were observed from 2009 in several municipalities of Liguria (Italy). Since the species is extremely polyphagous and is a real pest for several crops in tropical and subtropical areas, the presence of this alien insect is noteworthy, representing a new possible threat for native species and human activities.

The stereoselective synthesis of both enantiomers of trifluoro frontalin (-)-(1S,5R)- and (+)-(1R,5S)-8, as well as of diastereomeric monofluoro frontalines (-)-(1R,2R,5R)-18 and (-)-(1R,2S,5R)-20, analogues of the bioactive component of the aggregation pheromone of the Scolytidae insect family, has been accomplished starting from (-)-(1R)- and (+)-(1S)-menthyl (S)-toluene-4-sulfinate as a source of chirality and methyl trifluoroacetate or fluoroacetate, respectively, as sources of fluorine. The C-1 stereocenters were installed via stereoselective epoxidation of beta-sulfinyl ketones 2 and 13 with diazomethane. The bicyclic core was obtained by totally stereocontrolled and chemoselective tandem Wacker oxidation/intramolecular ketalization of the intermediate unsatured sulfinyl diols 5, 15, and 19. Axially fluorinated (-)-20 elicited a strong electroantennographic response in laboratory tests on females of Dendroctonus micans, whereas equatorially fluorinated (-)-18 and the trifluoroanalogue (-)-8 showed modest responses. Field trials using (-)-20 were not indicative owing to the locally scarce population of D. micans, but it showed some attractiveness for other Coleoptera families.