Bridging biotremology and chemical ecology: a new terminology.

Living organisms use both chemical and mechanical stimuli to survive in their environment. Substrate-borne vibrations play a significant role in mediating behaviors in animals and inducing physiological responses in plants, leading to the emergence of the discipline of biotremology. Biotremology is experiencing rapid growth both in fundamental research and in applications like pest control, drawing attention from diverse audiences. As parallels with concepts and approaches in chemical ecology emerge, there is a pressing need for a shared standardized vocabulary in the area of overlap for mutual understanding. In this article, we propose an updated set of terms in biotremology rooted in chemical ecology, using the suffix '-done' derived from the classic Greek word 'δονέω' (pronounced 'doneo'), meaning 'to shake'.

A multidisciplinary approach to tackling invasive species: barcoding, morphology, and metataxonomy of the leafhopper Arboridia adanae.

The leafhopper genus Arboridia includes several species that feed on Vitis vinifera and cause leaf chlorosis. We report the first alien Arboridia infestation in Italy in 2021 in an Apulian vineyard. To confirm the taxonomic status of the species responsible for crop damage, and reconstruct its demographic history, we barcoded individuals from Apulia together with Arboridia spp. from Crete (Greece), A. adanae from Central Turkey and other specimens of the presumed sister species, A. dalmatina from Dalmatia (Croatia). Molecular phylogenies and barcoding gap analysis identified clades not associated with sampling locations. This result is incongruent with classical specimen assignment and is further supported by morphological analyses, which did not reveal significant differences among the populations. Therefore, we propose A. dalmatina as a junior synonym of A. adanae, which would become the only grapevine-related Arboridia species in the eastern Mediterranean. To further characterise A. adanae evolution, we performed a molecular clock analysis that suggested a radiation during the Pleistocene glaciations. Finally, to assess whether the Apulian individuals carried microorganisms of agricultural relevance, we sequenced their bacterial microbiota using 16S rRNA amplicon sequencing identifying three phytopathogens not generally associated with Arboridia activities as well as Wolbachia in one Apulian haplogroup. We discuss the agricultural implications of this infestation.

The post-diapause vibrational behavior, motility, and survival of the brown marmorated stink bug Halyomorpha halys (Stål) adults at different temperatures.

Substrate-borne vibrational communication is common in pentatomids. Although several works exist on the vibrational communication of Halyomorpha halys, its vibrational behavior post diapause has not been investigated. In this study, we recorded H. halys overwintered adults using laser doppler vibrometers at three temperatures: 10 °C (inactivity), 18 °C (breaking of diapause), and 25 °C (peak of mating activity). The aim was to assess the effect of temperature on the signaling, motility, and survival of H. halys. The insects were sexed into different cages and recorded separately or joined with a cage of the opposite sex. We calculated the total time spent on signaling and walking per replica. The males predominantly emitted male signal 1 (MS1) throughout the four months of recordings. The females exclusively emitted female signal 2 (FS2) when joined with the opposite sex cage the first two months of recordings. Interestingly, they also started FS2 signaling when recorded separately, after two months. No signaling was recorded at 10 °C. At 25 °C, the signaling latency time before vibrational signaling was 24 h compared to 23 days at 18 °C. The short latency time at 25 °C correlated with a higher death rate in early stages of recording. Male walking activity was significantly higher in joined cages at 18 °C and 25 °C, suggesting the increased searching behavior near the opposite sex. Overwintered H. halys could adapt to different conditions whereas low temperatures maintain the diapause which is characterized by no signaling activity. Our results provide a foundation for bioclimatic modeling of climate change effects on H. halys and insights into the use of vibrational playbacks for mass trapping and monitoring as control techniques.

Plant protection and biotremology: fundamental and applied aspects.

There is overwhelming evidence that synthetic pesticides have a negative impact on the environment and human health, emphasizing the need for novel and sustainable methods for plant protection. A growing body of literature reports that plants interact through substrate-borne vibrations with arthropod pests and mutualistic arthropods that provide biological control and pollination services. Here, we propose a new theoretical framework that integrates insights from biological control, the ecology of fear, and plant-borne vibrations, to address plant-insect interactions and explore new, sustainable opportunities to improve plant health and productivity.

Effects of Irradiation on Biology and Mating Behaviour of Wild Males of Brown Marmorated Stink Bug Using a 6 MV Medical Linear Accelerator.

The brown marmorated stink bug, Halyomorpha halys, is a pentatomid bug of Eastern Asian origin that became an economically relevant pest in the Eurasian and American continents. Management of this species is limited to use of chemical insecticides: an inefficient method due to the strong adaptability of the target pest. The sterile insect technique (SIT) is potentially a valid tactic in the search for nontoxic alternatives. In this work, we investigated the suitability of mass-trapped overwintering males, collected during the aggregation phase before the winter diapause, for their release as competitive sterile males in an SIT programme. Differently from previous studies, irradiation was applied with a linear accelerator device that produced high-energy photons. Following a similar scientific protocol with newly emerged irradiated males, the effects of X-ray irradiation on physiological parameters (longevity, fecundity and fertility) were assessed. In addition, behavioural bioassays were carried out in no-choice conditions to evaluate if irradiation interferes with mating processes. The results are very encouraging; the effects of the irradiation at 32 Gy did not differ from the controls in the longevity or fecundity of the exposed overwintering adults. The hatching rate of the eggs laid by the fertile females that had mated with the irradiated males was less than 5%. The results of behavioural bioassays showed that the irradiation did not cause a significant impact on the quality of the sterile males. More research is warranted to evaluate the mating competitiveness of sterile males in semi-field and field conditions.

The Effect of the Sterile Insect Technique on Vibrational Communication: The Case of Bagrada hilaris (Hemiptera: Pentatomidae).

The painted bug, Bagrada hilaris, is an agricultural pest in its original areas (Africa, South Asia, and the Middle East), and it has recently been recorded as an invasive species in southwestern part of the US, Chile, Mexico, and two islands in the Mediterranean basin. Its polyphagous diet causes severe damage to economically important crops. The control of this pest is primarily achieved by means of synthetic pesticides, which are often expensive, ineffective, and harmful to the ecosystem. Recent physiological bioassays to assess its potential control through the sterile insect technique demonstrated that mating between untreated females and males irradiated at doses of 64 and 100 Gy, respectively, resulted in 90% and 100% sterility of the eggs produced by the females. In this study, the mating abilities of virgin males irradiated at 60 and 100 Gy with virgin females were measured through a study of short-range courtship mediated by vibrational communication. The results indicate that males irradiated at 100 Gy emit signals with lower peak frequencies, mate significantly less than unirradiated males do, and do not surpass the early stages of courtship. Conversely, males irradiated at 60 Gy present vibrational signal frequencies that are comparable to those of the control and successfully mated males. Our findings suggest that B. hilaris individuals irradiated at 60 Gy are good candidates for the control of this species, given that they retain sexual competitiveness regardless of their sterility, through an area-wide program that incorporates the sterile insect technique.

Orientus ishidae (Hemiptera: Cicadellidae): Biology, Direct Damage and Preliminary Studies on Apple Proliferation Infection in Apple Orchard.

The mosaic leafhopper, Orientus ishidae (Matsumura), is an Asian species widespread in Europe that can cause leaf damage in wild trees and transmit disease phytoplasmas to grapevines. Following an O. ishidae outbreak reported in 2019 in an apple orchard in northern Italy, the biology and damage caused by this species to apples were investigated during 2020 and 2021. Our studies included observations on the O. ishidae life cycle, leaf symptoms associated to its trophic activity, and its capability to acquire "Candidatus Phytoplasma mali," a causal agent of Apple Proliferation (AP). The results indicate that O. ishidae can complete the life cycle on apple trees. Nymphs emerged between May and June, and adults were present from early July to late October, with the peak of flight between July and early August. Semi-field observations allowed for an accurate description of leaf symptoms that appeared as a distinct yellowing after a one-day exposure. In field experiments, 23% of the leaves were found damaged. In addition, 16-18% of the collected leafhoppers were found carrying AP phytoplasma. We conclude that O. ishidae has the potential to be a new apple tree pest. However, further studies are required to better understand the economic impact of the infestations.

Factors influencing short-term parasitoid establishment and efficacy for the biological control of Halyomorpha halys with the samurai wasp Trissolcus japonicus.

BACKGROUND: Classical biological control has been identified as the most promising approach to limit the impact of the invasive pest species Halyomorpha halys (Heteroptera: Pentatomidae). This study investigated the parasitism rate at sites where the biocontrol agent Trissolcus japonicus (Hymenoptera: Scelionidae) was released and where its unintentional introduction took place, in the Trentino-South Tyrol region. The effect of land-use composition was studied to understand which factors favor the establishment of hosts and parasitoids, including native and exotic species. RESULTS: The released T. japonicus were detected a year after the start of the program, with a significant parasitoid impact and discovery, compared to control sites. Trissolcus japonicus was the most abundant H. halys parasitoid, and Trissolcus mitsukurii and Anastatus bifasciatus were recorded also. The efficacy of T. mitsukurii was lower in sites where T. japonicus was successfully established, suggesting a possible competitive interaction. Parasitism level by T. japonicus at the release sites was 12.5% in 2020 and 16.4% in 2021. The combined effect of predation and parasitization increased H. halys mortality up to 50% at the release sites. Landscape composition analysis showed that both H. halys and T. japonicus were more likely to be found at sites with lower altitude and with permanent crops, whereas other hosts and parasitoids preferred different conditions. CONCLUSION: Trissolcus japonicus showed a promising impact on H. halys, at release and adventive sites, with minor nontarget effects, mediated by landscape heterogeneity. The prevalence of T. japonicus in landscapes with permanent crops could support IPM in the future. © 2023 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Extending the vibroscape to agroecosystems: investigating the influence of abiotic factors and monitoring insect vibrational signaling.

Environmental conditions are crucial factors that influence communication systems and affect animal behavior. Research in the field of biotremology has improved our understanding of insect behavior, ecology, and evolution. However, the interactions between vibrational signaling and environmental factors are less studied, mainly because of technical issues faced in field trials. We therefore developed and tested an approach to investigate the effect of abiotic factors on insect vibrational signaling and explored its implementation as a monitoring tool for insect vibrational signals, using a vineyard as an agroecosystem model. Our results showed a significant decrease in insect signaling activity during unsuitable conditions of high temperature and wind velocity. We determined for the first time, the daily signaling pattern of the two insect pests, Scaphoideus titanus and Halyomorpha halys, in natural conditions. Biotremology techniques could be profitably used to monitor not only the presence of target pest species but also the biodiversity associated with vibrational signaling insects. In particular, the method implemented in this study could be used as a tool to compare the quality of cultivated areas under different management systems.

Coupling computational vibrational models and experimental biotremology to develop a green pest control strategy against the greenhouse whitefly Trialeurodes vaporariorum.

In applied biotremology, vibrational signals or cues are exploited to manipulate the target species behaviour. To develop an efficient pest control strategy, other than a detailed investigation into the pest biology and behaviour, the role of the substrate used to transmit the signal is an important feature to be considered, since it may affect vibrations spreading and effective signal transmission and perception. Therefore, we used a multi-disciplinary approach to develop a control technique against the greenhouse whitefly, Trialeurodes vaporariorum. First, an ad hoc vibrational disruptive noise has been developed, based on the acquired knowledge about the mating behaviour and vibrational communication of the mated species. Subsequently, we employed finite-element models to investigate a growing tomato plant response to the aforesaid noise. Modelling how vibrations spread along the plant allowed us to set up a greenhouse experiment to assess the efficacy in terms of insect population of the vibrational treatment, which was administrated through vibrational plates. The green methodology applied in this study represents an innovative, environmentally sound alternative to the usage of synthetic pesticides.

Effects of Gamma Irradiation on the Fecundity, Fertility, and Longevity of the Invasive Stink Bug Pest Bagrada hilaris (Burmeister) (Hemiptera: Pentatomidae).

The bagrada bug, Bagrada hilaris, is an invasive insect pest in the family Brassicaceae that causes economically important damage to crops. It was originally present in Asia, the Middle East, and Africa, and was reported as invasive in the southwestern part of the US, in Chile, and on a few islands in the Mediterranean Basin. In its native range, B. hilaris is controlled by several egg parasitoid species that are under consideration as potential biological control agents. This research evaluated the impact of gamma irradiation on life history parameters, e.g., the fecundity, fertility, and longevity of B. hilaris, as a critical step towards assessing the feasibility of using the sterile insect technique against this recent invasive pest. Newly emerged adults of a laboratory colony originally collected from the island of Pantelleria (Italy) were gamma-irradiated. Life history parameters were evaluated at nine different doses, ranging from 16 Gy to 140 Gy. The minimal dose to approach full sterility was 100 Gy. Irradiation up to a maximum of 140 Gy apparently did not negatively impact the longevity of the adults. Even if both genders are sensitive to irradiation, the decline in fecundity for irradiated females could be exploited to release irradiated males safely to apply the SIT in combination with classical biological control. The data presented here allow us to consider, for the first time, the irradiation of bagrada adults as a suitable and feasible technique that could contribute to guaranteeing a safe approach to control this important pest species in agro-ecosystems. More research is warranted on the competitive fitness of irradiated males to better understand mating behavior as well as elucidate the possible mechanisms of sperm selection by polyandric B. hilaris females.

Application of vibrational signals to study and manipulate an insect vector: the case of Philaenus spumarius (Hemiptera: Aphrophoridae).

BACKGROUND: Vibrational stimuli can support pest management as they provide environmentally friendly methods to manipulate insect pest behaviors. Different vibrational stimuli were used to study and influence the behavior of the meadow spittlebug, Philaenus spumarius, the European vector of Xylella fastidiosa. In playback experiments, we tested the reactions of the spittlebug toward the male calling signals (test 1) and the male-male signal (test 2). In test 3, we evaluated the use of conspecific signals and noises to repel insects/disrupt mating. RESULTS: Test 1 provided new insights regarding the role of the male calling signal in intraspecific communication, in particular that this signal likely does not underlie aggregation or aggression toward conspecifics. Test 2 demonstrated that the male-male signal is used by males to express distress when physically interacting, whilst, when played back into a host plant, it has not any repellent effect on the spittlebug. Test 3A suggested that males exploit short silence gaps to localize the signaling partner, while test 3B showed that a continuous noise with a specific frequency range successfully disrupt mating, as only one male out of 20 localized the female on the plant. CONCLUSION: Playbacks obtained from prerecorded P. spumarius' signals were successfully used to accomplish ethological studies; even so, this approach did not show a real potential to be used as a control strategy. However, noises designed to mask the spittlebug signals significantly disrupted species mating and could integrate other techniques aimed at reducing the spread of X. fastidiosa after appropriate implementation. © 2022 Society of Chemical Industry.

Effect of vibrational mating disruption on flight activity and oviposition to control the grapevine pest, Scaphoideustitanus.

The increasing demand for safe and sustainably produced food is leading to the development of strategies of pest control alternative to chemicals. One innovative method is Vibrational Mating Disruption (VMD) to disrupt insect communication in plants. VMD was proven effective in preventing mating of the grapevine pest Scaphoideus titanus, vector of flavescence dorée. However, the stress induced by VMD on the target species has the potential to influence other crucial aspects of the insect biology and ethology. Therefore, the goal of this study was to understand side effects of VMD on the flight activity and oviposition of S. titanus. The results of our experiments conducted in the greenhouse showed that in the presence of a receptive female, males fly more if exposed to vibrations than in the silent control but not differently from singles males in silence. Surprisingly, we found that also females subjected to VMD fly more than in the silence. Regarding oviposition, we found that mated females exposed to vibrations and single females (unmated) laid significantly fewer eggs than mated females in silence. In conclusion, this study shows the potential of VMD to interfere, besides with mating, with other important biological aspects of the pest species.

Attraction of Egg Parasitoids Trissolcus mitsukurii and Trissolcus japonicus to the chemical cues of Halyomorpha halys and Nezara viridula.

Trissolcus mitsukurii and Trissolcus japonicus are two Asian egg parasitoids associated with different pentatomids such as Halyomorpha halys. Adventive populations of T. mitsukurii were found in Northern Italy, suggesting its employment as a biological control agent (BCA) against H. halys. Nevertheless, to reduce the latter's population, T. japonicus was released in Italy. Releasing an exotic parasitoid requires investigating the interaction between the BCA and the environment to avoid negative impacts on the entomofauna of the new habitat. Trissolcus mitsukurii is mainly associated with Nezara viridula in its native area. Therefore, we investigated and compared the ability of female T. mitsukurii and T. japonicus to distinguish between naturally released cues of H. halys and N. viridula. A single parasitoid was exposed to contact kairomones of both pests to evaluate its modifications in orthokinetic and locomotory behaviour. The behaviour of female T. mitsukurii was also tested on synthetic compounds simulating the cues of the two pentatomids. When naturally released cues were used, T. japonicus preferred the traces of H. halys, while T. mitsukurii preferred N. viridula's cues. Moreover, the attraction of T. mitsukurii to N. viridula's cues was confirmed with synthetic cues. Additional studies are needed to judge if this parasitoid can be used as a BCA.

Design of ideal vibrational signals for stinkbug male attraction through vibrotaxis experiments.

BACKGROUND: Many groups of insects utilize substrate-borne vibrations for intraspecific communication. This characteristic makes them a suitable model for exploring the use of vibrations as a tool for pest control as an alternative to the use of chemicals. Detailed knowledge of species communication is a prerequisite to select the best signals to use. This study explored the use of substrate-borne vibrations for pest control of the brown marmorated stink bug (BMSB), Halyomorpha halys Stål (Heteroptera: Pentatomidae). For this purpose, we first identified the spectral and temporal characteristics that best elicit male responsiveness. Bioassays were conducted with artificial signals that mimicked the natural female calling signal. Second, we used the acquired knowledge to synthesize new signals endowed with different degrees of attractiveness in single- and two-choice bioassays using a wooden custom-made T stand. RESULTS: The results from this study showed that males were attracted to female signals along a high range of amplitudes, especially starting from a threshold of 100 µm s-1 , a high pulse repetition time (1 s) and frequency peak corresponding to the first harmonic (76 Hz). This resulted in an "optimal" signal for use to attract males, while the choice test in the T arena showed that this signal elicits searching behavior and attracts BMSB males towards a stimulation point. CONCLUSION: We confirm the use of vibrational signals as a strong tool for behavioral manipulation of male BMSB and suggest its possible use in the development of field traps and further management of this pest. © 2021 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Intrasexual Vibrational Behavior of Philaenus spumarius in Semi-Field Conditions.

Insects that communicate by vibrational signals live in a complex interactive network of communication. Most studies on insect intrasexual behavior, based on plant-borne vibrational signals, have targeted few individuals. Despite their importance, behaviors that occur within groups were often overlooked. The study of multiple individuals, when insects occur in high density could simulate the environment in which they live and provide more reliable information on their behavior. In semi-field conditions, we investigated the intrasexual behavior of the meadow spittlebug, Philaenus spumarius. Vibrational signals exchanged among individuals of the same sex were recorded throughout their adult stage, from late spring to early autumn, and during the day, from the morning to the evening using a laser vibrometer. Males were less active than females throughout the season and their interactions were less frequent compared to females. Intrasexual interactions were characterized by signal overlapping in both unisex groups, in addition to signal alternating only in the case of males. In conclusion, the study of signaling behavior in intrasexual groups contributed to a better understanding of P. spumarius social behavior. We discuss the hypothesis of a possible competitive behavior between males and cooperative behavior between females.

Beneficial Insects Deliver Plant Growth-Promoting Bacterial Endophytes between Tomato Plants.

Beneficial insects and mites, including generalist predators of the family Miridae, are widely used in biocontrol programs against many crop pests, such as whiteflies, aphids, lepidopterans and mites. Mirid predators frequently complement their carnivore diet by feeding plant sap with their piercing-sucking mouthparts. This implies that mirids may act as vectors of phytopathogenic and beneficial microorganisms, such as plant growth-promoting bacterial endophytes. This work aimed at understanding the role of two beneficial mirids (Macrolophus pygmaeus and Nesidiocoris tenuis) in the acquisition and transmission of two plant growth-promoting bacteria, Paraburkholderia phytofirmans strain PsJN (PsJN) and Enterobacter sp. strain 32A (32A). Both bacterial strains were detected on the epicuticle and internal body of both mirids at the end of the mirid-mediated transmission. Moreover, both mirids were able to transmit PsJN and 32A between tomato plants and these bacterial strains could be re-isolated from tomato shoots after mirid-mediated transmission. In particular, PsJN and 32A endophytically colonised tomato plants and moved from the shoots to roots after mirid-mediated transmission. In conclusion, this study provided novel evidence for the acquisition and transmission of plant growth-promoting bacterial endophytes by beneficial mirids.

Behavioral Manipulation for Pest Control.

Pest control is moving towards a dramatic reduction in pesticide-based approaches in favor of more eco-friendly strategies characterized by the promotion of ecological intensification of agriculture and reduction of human inputs (especially pesticides) [...].

Vibrational communication and mating behavior of the greenhouse whitefly Trialeurodes vaporariorum (Westwood) (Hemiptera: Aleyrodidae).

The greenhouse whitefly (GW), Trialeurodes vaporariorum is considered one of the most harmful insect pests in greenhouses worldwide. The GW mating behavior has been partially investigated and its vibrational communication is only in part known. A deeper knowledge of its intraspecific communication is required to evaluate the applicability of control methods based on techniques of behavioral manipulation. In this study, for the first time, we provided a detailed ethogram of the GW mating behavior and we characterized the vibrational signals emitted during the process of pair formation. We characterized two types of male vibrational emissions ("chirp" and "pulses"), differently arranged according to the behavioral stage to form stage-specific signals, and a previously undescribed Male Rivalry Signal. We recorded and characterized two new female signals: The Female Responding Signal and the Female Rejective Signal. The mating behavior of GW can be divided into six different stages that we named "call", "alternated duet", "courtship", "overlapped duet", "mating", "failed mating attempt". The analysis performed with the Markovian behavioral transition matrix showed that the "courtship" is the key stage in which male exhibits its quality and can lead to the "overlapped duet" stage. The latter is strictly associated to the female acceptance and therefore it plays a crucial role to achieve mating success. Based on our findings, we consider the use of vibrational playbacks interfering with GW mating communication a promising option for pest control in greenhouses. We discuss the possibility to start a research program of behavioral manipulation to control the populations of GW.