A model for predicting the phenology of Philaenus spumarius.

The design and implementation of Philaenus spumarius control strategies can take advantage of properly calibrated models describing and predicting the phenology of vector populations in agroecosystems. We developed a temperature-driven physiological-based model based on the system of Kolmogorov partial differential equations to predict the phenological dynamics of P. spumarius. The model considers the initial physiological age distribution of eggs, the diapause termination process, and the development rate functions of post-diapausing eggs and nymphal stages, estimated from data collected in laboratory experiments and field surveys in Italy. The temperature threshold and cumulative degree days for egg diapause termination were estimated as 6.5 °C and 120 DD, respectively. Preimaginal development rate functions exhibited lower thresholds ranging between 2.1 and 5.0 °C, optimal temperatures between 26.6 and 28.3 °C, and upper threshold between 33.0 and 35 °C. The model correctly simulates the emergence of the 3rd, 4th, and 5th nymphal instars, key stages to target monitoring actions and control measures against P. spumarius. Precision in simulating the phenology of the 1st and 2nd nymphal stages was less satisfactory. The model is a useful rational decision tool to support scheduling monitoring and control actions against the late and most important nymphal stages of P. spumarius.

Activity of natural occurring entomopathogenic fungi on nymphal and adult stages of Philaenus spumarius.

The spittlebug Philaenus spumarius (Hemiptera: Aphrophoridae) is the predominant vector of Xylella fastidiosa (Xanthomonadales: Xanthomonadaceae) in Apulia, Italy and the rest of Europe. Current control strategies of the insect vector rely on mechanical management of nymphal stages and insecticide application against adult populations. Entomopathogenic fungi (EPF) are biological control agents naturally attacking spittlebugs and may effectively reduce population levels of host species. Different experimental trials in controlled conditions have been performed to i) identify naturally occurring EPF on P, spumarius in Northwestern Italy, and ii) evaluate the potential for biocontrol of the isolated strains on both nymphal and adult stages of the spittlebug. Four EPF species were isolated from dead P. spumarius collected in semi-field conditions: Beauveria bassiana, Conidiobolus coronatus, Fusarium equiseti and Lecanicillium aphanocladii. All the fungal isolates showed entomopathogenic potential against nymphal stages of P. spumarius (≈ 45 % mortality), except for F. equiseti, in preliminary trials. No induced mortality was observed on adult stage. Lecanicillium aphanocladii was the most promising fungus and its pathogenicity against spittlebug nymphs was further tested in different formulations (conidia vs blastospores) and with natural adjuvants. Blastospore formulation was the most effective in killing nymphal instars and reducing the emergence rate of P, spumarius adults, reaching mortality levels (90%) similar to those of the commercial product Naturalis®, while no or adverse effect of natural adjuvants was recorded. The encouraging results of this study pave way for testing EPF isolates against P, spumarius in field conditions and find new environmentally friendly control strategies against insect vectors of X. fastidiosa.

Competition among Flavescence Dorée Phytoplasma Strains in the Experimental Insect Vector Euscelidius variegatus.

Phytoplasmas are plant pathogenic wall-less bacteria transmitted in a persistent propagative manner by hemipteran insects, mainly belonging to the suborder Auchenorrhyncha (Fulgoromorpha and Cicadomorpha). Flavescence dorée (FD) is a quarantine disease of grapevine, causing great damage to European viticulture and associated with phytoplasmas belonging to 16SrV-C (FD-C) and -D (FD-D) subgroups. FD-C and FD-D strains share similar pathogenicity, but mixed infections are rare in nature. To investigate the competition among FDp strains, specimens of the laboratory vector Euscelidius variegatus (Hemiptera: Cicadellidae) were forced to acquire both phytoplasma haplotypes upon feeding on FD-C- and FD-D-infected plants or after the injection of both strains. The pathogen colonization of insect bodies and heads was monitored with multiplex qPCR, and the efficiencies of phytoplasma transmission were estimated. Single infection, irrespective of strain type, was more frequent than expected, indicating that competition among FD strains occurs. Hypotheses of competition for resources and/or host active sites or the direct antibiosis of one strain against the other are discussed, based on the genetic complexity of FDp populations and on the high genome variability of the FD-D strain. As FD management still mainly relies on insecticides against vectors, the characterization of FDp haplotypes and the description of their epidemiology also have practical implications.

Bioecological Traits of Spittlebugs and Their Implications for the Epidemiology and Control of the Xylella fastidiosa Epidemic in Apulia (Southern Italy).

Spatial-temporal dynamics of spittlebug populations, together with transmission biology, are of major importance to outline the disease epidemiology of Xylella fastidiosa subsp. pauca in Apulian olive groves. The spread rate of X. fastidiosa is mainly influenced by (i) the pathogen colonization of the host plant; (ii) the acquisition of the pathogen by the vector from an infected plant, and its inoculation to healthy plants; (iii) the vector population dynamics and abundance at different spatial scales; and (iv) the dispersal of the vector. In this contribution we summarize the recent advances in research on insect vectors' traits-points ii, iii, and iv-focusing on those most relevant to X. fastidiosa epidemic in Apulia. Among the vectors' bioecological traits influencing the X. fastidiosa epidemic in olive trees, we emphasize the following: natural infectivity and transmission efficiency, phenological timing of both nymphal and adult stage, the role of seminatural vegetation as a vector reservoir in the agroecosystem and landscape, and preferential and directional dispersal capabilities. Despite the research on X. fastidiosa vectors carried out in Europe in the last decade, key uncertainties on insect vectors remain, hampering a thorough understanding of pathogen epidemiology and the development of effective and targeted management strategies. Our goal is to provide a structured and contextualized review of knowledge on X. fastidiosa vectors' key traits in the Apulian epidemic, highlighting information gaps and stimulating novel research pathways on X. fastidiosa pathosystems in Europe. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Transmission of Xylella fastidiosa subsp. pauca ST53 by the Sharpshooter Cicadella viridis From Different Source Plants and Artificial Diets.

The sharpshooter Cicadella viridis L. (Hemiptera: Cicadellidae) is the most common sharpshooter in Europe and, given its xylem feeding behavior, is considered a potential vector of the plant pathogenic bacterium Xylella fastidiosa Wells et al. (Xanthomonadales: Xanthomonadaceae). We tested X. fastidiosa subsp. pauca ST53 (Xfp) transmission capabilities of C. viridis adults, namely 1) acquisition efficiency from four host plant species-periwinkle, milkwort, lavender, alfalfa-and from two artificial diets (PD3 and Xfm), 2) inoculation efficiency to periwinkle at different times post acquisition from different plant and artificial diet sources. The main European vector species-Philaenus spumarius L. (Hemiptera: Aphrophoridae)-was used as a control. C. viridis was able to acquire Xfp from periwinkle, milkwort, and lavender, although with low efficiency (3-16%) and from artificial diets (23-25%). Successful inoculation on periwinkle was extremely rare, being observed only three times, following feeding on milkwort plant and PD3 artificial diet sources. Our study shows that C. viridis is not a relevant vector of Xfp, given the very low transmission rate in controlled conditions, and the inability to feed on olive. The low efficiency reported here correlates with ecological constraints of the vector (mainly monocots host plants, humid environments) that make it difficult to forecast a relevant role in dispersing X. fastidiosa, at least within the present distribution of the exotic bacterium in Europe. However, a possible role of this species in spreading Xf in other agroecosystems, e.g., vineyard and stone fruits grown in humid areas, cannot be excluded.

Phenology, Seasonal Abundance, and Host-Plant Association of Spittlebugs (Hemiptera: Aphrophoridae) in Vineyards of Northwestern Italy.

Spittlebugs (Hemiptera: Aphrophoridae) are the vectors of the bacterium Xylella fastidiosa (Xf) in Europe. Xf may cause severe epidemics in cultivated plants, including grapevines. To assess the threat represented by the bacterium to grapevines, detailed information on the vectors' phenology, density, and ecology in vineyards is needed. The aim of the present work was to describe spittlebug diversity, phenology, and host-plant association in the vineyard agroecosystem. Two separate field surveys of nymphal and adult spittlebug populations, i.e., a two-year survey of a single site and a one-year survey of three sites, were performed in vineyards of northwestern Italy in three consecutive years. Philaenus spumarius was the most common species, reaching average nymph densities on herbaceous cover up to 60-130 nymphs/m2. Adults were sampled on grapevines from May to September, with a peak in June (up to 0.43 insects/sweep). Herbaceous cover was colonized after egg hatching and in late summer for oviposition, while wild woody hosts represented a refuge during summer. The results show that spittlebugs can reach high population levels in vineyards, at least in the areas where the ground is covered by herbaceous plants for the whole season and the use of insecticides is moderate. The extended presence of P. spumarius adults on grapevines represents a serious risk factor for the spread of Xf. The scenarios of Xf establishment in vineyards in northwestern Italy and Europe are discussed in relation to the abundance, phenology, and plant association of spittlebugs.

Dispersal of Philaenus spumarius (Hemiptera: Aphrophoridae), a Vector of Xylella fastidiosa, in Olive Grove and Meadow Agroecosystems.

The introduction of the Xylella fastidiosa Wells bacterium into Apulia (South Italy) has caused the massive dieback of olive trees, and is threatening olive production throughout the Mediterranean Region. The key vector of X. fastidiosa in Europe is the spittlebug Philaenus spumarius L. The dispersal capabilities of P. spumarius are poorly known, despite being a key parameter for the prediction of the spread of the bacterium. In this study, we have examined the dispersal of P. spumarius adults in two different agroecosystems in Italy: an olive grove in Apulia (Southern Italy) and a meadow in Piedmont (Northern Italy). Insects were marked with albumin and released during seven independent trials over 2 yr. The recapture data were pooled separately for each agroecosystem and used to estimate the dispersal kernels of P. spumarius in the olive grove and in the meadow. The diffusion coefficient estimate for P. spumarius was higher in the meadow than in the olive grove. The median distance from the release point for 1 d of dispersal was 26 m in the olive grove and 35 m in the meadow. On the basis of our model, we estimated that 50% of the spittlebug population remained within 200 m (98% within 400 m) during the 2 mo period of high abundance of the vector on olives in Apulia. The dispersal of P. spumarius is thus limited to some hundreds of meters throughout the whole year, although it can be influenced to a great extent by the structure of the agroecosystem.

Biology and Prevalence in Northern Italy of Verrallia aucta (Diptera, Pipunculidae), a Parasitoid of Philaenus spumarius (Hemiptera, Aphrophoridae), the Main Vector of Xylella fastidiosa in Europe.

The meadow spittlebug, Philaenus spumarius (L.) (Hemiptera Aphrophoridae), the main vector of Xylella fastidiosa Wells et al. in Europe, has few known natural enemies. The endoparasitoid Verrallia aucta (Fallén) (Diptera, Pipunculidae) was first noticed a long time ago but very little is known about its biology and prevalence. In this study, the presence and prevalence of V. aucta were investigated in different regions of northern Italy, both in plain-foothill and montane zones. Parasitic larvae were identified by the dissection of spittlebug adults, P. spumarius and Neophilaenus campestris (Fallén), and by a new species-specific molecular tool targeting the ITS2 and COI genomic regions, developed in this work. A small-scale rearing was set up to gain information on the life cycle of V. aucta on its main host P. spumarius. During the four-year investigation (2016-2019) the pipunculid parasitoid displayed low prevalence, reaching a maximum parasitization rate of 17.5% (calculated over the adult spittlebug season) in vineyards of the Piemonte region. Over the whole period, no significant difference in the prevalence was found between male and female spittlebugs. Collected data and rearing observations suggest that V. aucta is monovoltine and synchronous with P. spumarius, laying eggs in newly emerged adults, developing as an endoparasitoid through two larval stages during the whole summer, and overwintering as a pupa in the soil.

Prevalence of Flavescence Dorée Phytoplasma-Infected Scaphoideus titanus in Different Vineyard Agroecosystems of Northwestern Italy.

Quantitative estimates of vector populations and their infectivity in the wild and in cultivated compartments of agroecosystems have been carried out to elucidate the role of the wild compartment in the epidemiology of Flavescence dorée (FD). Seven sites were selected for the investigations in the Piedmont Region of Italy. They were characterized by a high variety of agricultural and ecological landscape features, and included a vineyard surrounded by wild vegetation. In order to describe abundance and prevalence of FD-infected vectors in the cultivated and wild compartments of the vineyard agroecosystem, adults of Scaphoideus titanus were collected by yellow sticky traps inside and outside the vineyard over the period July 10th-September 9th, 2015. They were counted and singly analyzed for the presence of FD phytoplasmas by PCR. Multifactorial correlations among vector population level, prevalence of infected insects inside and outside the vineyards, disease prevalence in cultivated and wild Vitis plants, and location of wild Vitis plants with respect to the vineyard were analyzed. Abundance of S. titanus adults significantly decreased from the end of July onwards, particularly inside the vineyard (average range 22.7 ± 2.5 insects/trap). Percentage of FD-positive S. titanus was significantly higher outside the vineyard (up to 48% on average) compared to inside the vineyard (up to 34% on average), and increased during the season in both compartments.

Spittlebugs of Mediterranean Olive Groves: Host-Plant Exploitation throughout the Year.

Spittlebugs are the vectors of the bacterium Xylella fastidiosa Wells in Europe, the causal agent of olive dieback epidemic in Apulia, Italy. Selection and distribution of different spittlebug species on host-plants were investigated during field surveys in 2016-2018 in four olive orchards of Apulia and Liguria Regions of Italy. The nymphal population in the herbaceous cover was estimated using quadrat samplings. Adults were collected by sweeping net on three different vegetational components: herbaceous cover, olive canopy, and wild woody plants. Three species of spittlebugs were collected: Philaenus spumarius L., Neophilaenus campestris (Fallén), and Aphrophora alni (L.) (Hemiptera: Aphrophoridae). Philaenus spumarius was the predominant species both in Apulia and Liguria olive groves. Nymphal stages are highly polyphagous, selecting preferentially Asteraceae Fabaceae plant families, in particular some genera, e.g., Picris, Crepis, Sonchus, Bellis, Cichorium, and Medicago. Host-plant preference of nymphs varies according to the Region and through time and nymphal instar. In the monitored sites, adults peak on olive trees earlier in Apulia (i.e., during inflorescence emergence) than in Liguria (i.e., during flowering and beginning of fruit development). Principal alternative woody hosts are Quercus spp. and Pistacia spp. Knowledge concerning plant selection and ecological traits of spittlebugs in different Mediterranean olive production areas is needed to design effective and precise control strategies against X. fastidiosa vectors in olive groves, such as ground cover modifications to reduce populations of spittlebug vectors.

Phenology, seasonal abundance and stage-structure of spittlebug (Hemiptera: Aphrophoridae) populations in olive groves in Italy.

Spittlebugs (Hemiptera: Aphrophoridae) are the dominant xylem-sap feeders in the Mediterranean area and the only proven vectors of Xylella fastidiosa ST53, the causal agent of the olive dieback epidemic in Apulia, Italy. We have investigated the structured population phenology, abundance and seasonal movement between crops and wild plant species of both the nymphal and adult stages of different spittlebug species in olive groves. Field surveys were conducted during the 2016-2018 period in four olive orchards located in coastal and inland areas in the Apulia and Liguria regions in Italy. The nymphal population in the herbaceous cover was estimated using quadrat samplings. Adults were collected through sweep nets on three different vegetational components: herbaceous cover, olive canopy and wild woody plants. Philaenus spumarius was the most abundant species; its nymphs were collected from early March and reached a peak around mid-April, when the 4th instar was prevalent. Spittlebug adults were collected from late April until late autumn. P. spumarius adults were abundant on the herbaceous cover and olive trees in late spring, and they then dispersed to wild woody hosts during the summer and returned to the olive groves in autumn when searching for oviposition sites in the herbaceous cover. A relatively high abundance of P. spumarius was observed on olive trees during summer in the Liguria Region. The present work provides a large amount of data on the life cycle of spittlebugs within an olive agroecosystem that can be used to design effective control programmes against these vectors in infected areas and to assess the risk of the establishment and spread of X. fastidiosa to Xylella-free areas.

Plant Selection and Population Trend of Spittlebug Immatures (Hemiptera: Aphrophoridae) in Olive Groves of the Apulia Region of Italy.

The xylem-limited bacterium Xylella fastidiosa Wells is the causal agent of severe diseases of several cultivated and wild plants. It is transmitted by xylem-sap feeder insects, such as spittlebugs (Hemiptera: Cercopoidea) and sharpshooters (Hemiptera: Cicadellinae). A dramatic epidemic of X. fastidiosa subspecies pauca sequence type 53 is currently affecting a large area of the Apulia Region of Italy, where it is spread by Philaenus spumarius L. adults within olives. In 2015 and 2016, field surveys were carried out in Apulian olive groves to investigate host plant selection of spittlebug nymphs, to identify the main plant species that can act as reservoirs of the vectors. Two different sampling methods were used: randomized plant sampling and quadrats sampling. Host plant selection by P. spumarius and Neophilaenus campestris (Fallén) nymphs was estimated using Manly's selection index. The botanic families presenting the highest number of plants infested by P. spumarius nymphs were Asteraceae, Fabaceae, and Apiaceae. Nymphs of P. spumarius were sampled on 72 plant genera, and among the most common 25 genera, Sonchus, Knautia, Glebionis, Urospermum (Asteraceae), Medicago, Vicia, Melilotus (Fabaceae), and Daucus (Apiaceae) were the ones selected preferentially, according to Manly's index results. Populations of P. spumarius nymphs peak in early April, with densities ranging between 10 and 40 nymph/m2, were about 10-fold those of N. campestris. Plant infestation rate by spittlebug nymphs in 2016 was significantly higher in olive groves located in Lecce province (infected area) than those situated in Bari province (noninfected area).