Bioactivity of Carlina acaulis Essential Oil and Its Main Component towards the Olive Fruit Fly, Bactrocera oleae: Ingestion Toxicity, Electrophysiological and Behavioral Insights.

Among botanical insecticides based on essential oils (EOs) or their main components, Carlina acaulis EO and the aromatic polyacetylene carlina oxide, constituting more than 90% of its EO, were recently proven to be effective against the larvae and adults of some insect vectors and pests. In this study, the toxicity of C. acaulis EO and carlina oxide were tested on Bactrocera oleae adults using a protein bait formulation. The LC50 values of the C. acaulis EO and carlina oxide were 706 ppm and 1052 ppm, respectively. Electroantennographic (EAG) tests on B. oleae adults showed that both carlina EO and oxide elicited EAG dose-dependent responses in male and female antennae. The responses to the EO were significantly higher than those to carlina oxide, indicating that other compounds, despite their lower concentrations, can play a relevant role. Moreover, Y-tube assays carried out to assess the potential attractiveness or repellency of carlina oxide LC90 to B. oleae adults showed that it was unattractive to both males and females of B. oleae, and the time spent by both sexes in either the control or the treatment arm did not differ significantly. Overall, this study points out the potential use of C. acaulis EO and carlina oxide for the development of green and effective "lure-and-kill" tools.

Bioactivity of Wild Hop Extracts against the Granary Weevil, Sitophilus granarius (L.).

The use of bioinsecticides, rather than synthetic compounds, appears a goal to be pursued in pest control, especially for species such as Sitophilus granarius (L.) which attack stored products. Since Humulus lupulus (L.) is a remarkable source of bioactive compounds, this study investigated the bioactivity of hop flower extracts against S. granarius adults by evaluating toxic (contact, inhalation, and ingestion), repellent, antifeedant, and nutritional effects as well as their anticholinesterase activity and olfactory sensitivity. Hop extracts were obtained by soaking dried and ground hop cones in solvents of decreasing polarity: methanol, acetone, and n-hexane. Dried crude extracts were resuspended in each solvent, and used in topical application, ingestion, and fumigation toxicity assays, as well as in contact and short-range repellency tests, in vitro anticholinesterase activity evaluation, and electroantennographic tests. No inhalation toxicity for the extracts was found. On the contrary, all extracts showed adult contact toxicity 24 h after treatment (LD50/LD90 16.17/33.20, 25.77/42.64, and 31.07/49.48 µg/adult for acetone, n-hexane, and methanol extracts, respectively); negligible variations for these values at 48 h were found. The anticholinesterase activity shown by all extracts suggested that the inhibition of this enzyme was one of the mechanisms of action. Interestingly, flour disk bioassays revealed a significant ingestion toxicity for the acetone extract and a lower toxicity for the other two extracts. Moreover, all extracts affected insect nutritional parameters, at the highest dose checked. Filter paper and two-choice pitfall bioassays showed repellent activity and a strong reduction of insect orientation to a highly attractive food odor source, with minor differences among extracts, respectively. Finally, the presence of volatile compounds in the different extracts that are perceived by insect antennae was confirmed by electroantennography. All these findings strongly suggest a possible use of hop cone extracts against S. granarius, thus further confirming this plant as an interesting species for pest control.

Biological Activity of Humulus Lupulus (L.) Essential Oil and Its Main Components Against Sitophilus granarius (L.).

Besides its use in the brewing industry, hop cones appear as a powerful source of biologically active compounds, already checked for their putative anticancer, antimicrobial, and other bioactivities. Conversely, hop use in pest control remains to date under-investigated. Therefore, the biological activity of hop essential oil (EO) and its main constituents was investigated here against Sitophilus granarius. Adult contact toxicity was found 24 h after treatment with hop EO (LD50/LD90 13.30/40.23 µg/adult), and its three most abundant components, α-humulene, ß-myrcene, and ß-caryophyllene (LD50/LD90 41.87/73.51, 75.91/126.05, and 138.51/241.27 µg/adult, respectively); negligible variations at 48 h, except for α-humulene (LD50/LD90 26.83/49.49 µg/adult), were found. The fumigant toxicity of the EO and terpenes was also checked: in the absence of wheat grains, ß-myrcene showed the highest inhalation toxicity (LC50/LC90 72.78/116.92 mg/L air), whereas α-humulene, ß-caryophyllene, and the EO induced similar values (LC50/LC90 about 130/200 mg/L air); with the exception for EO, the wheat presence increased (30-50%) LC50/LC90 values. Moreover, EO and terpenes were perceived by insect antennae and elicited repellent activity. Only ß-caryophyllene showed an anticholinesterase effect, this suggesting that different mechanisms of action should be responsible for hop EO toxicity. Therefore, hop EO appears suitable for developing control means against this pest.

Electrophysiological and behavioural response of Philaenus spumarius to essential oils and aromatic plants.

The meadow spittlebug, Philaenus spumarius, is a highly polyphagous widespread species, playing a major role in the transmission of the bacterium Xylella fastidiosa subspecies pauca, the agent of the "Olive Quick Decline Syndrome". Essential oils (EOs) are an important source of bio-active volatile compounds that could interfere with basic metabolic, biochemical, physiological, and behavioural functions of insects. Here, we report the electrophysiological and behavioural responses of adult P. spumarius towards some EOs and related plants. Electroantennographic tests demonstrated that the peripheral olfactory system of P. spumarius females and males perceives volatile organic compounds present in the EOs of Pelargonium graveolens, Cymbopogon nardus and Lavandula officinalis in a dose-dependent manner. In behavioral bioassays, evaluating the adult responses towards EOs and related plants, both at close (Y-tube) and long range (wind tunnel), males and females responded differently to the same odorant. Using EOs, a clear attraction was noted only for males towards lavender EO. Conversely, plants elicited responses that varied upon the plant species, testing device and adult sex. Both lavender and geranium repelled females at any distance range. On the contrary, males were attracted by geranium and repelled by citronella. Finally, at close distance, lavender and citronella were repellent for females and males, respectively. Our results contribute to the development of innovative tools and approaches, alternative to the use of synthetic pesticides, for the sustainable control of P. spumarius aiming to contrasting the expansion of X. fastidiosa.

Electroantennographic Responses of Aromia bungii (Faldermann, 1835) (Coleoptera, Cerambycidae) to a Range of Volatile Compounds.

BACKGROUND: The red-necked longhorn beetle, Aromia bungii, is one of the most damaging pests of stone fruit trees. Native to the south-eastern Palearctic and Oriental regions, it invaded and is established to some extent in the Campania Region (Southern Italy). In several cerambycid species, volatile organic compounds (VOCs) have been shown to play a role in mate and host plant location. METHODS: The electroantennographic (EAG) technique was employed to explore the antennal chemoreceptivity of male and female A. bungii antennae to 90 VOCs. RESULTS: Increasing EAG amplitudes from the basal to the distal antennal segments were recorded in response to six selected plant volatiles. From the distal flagellomeres, the largest EAG responses (>0.8 mV) were elicited by 2-hexanol, octanal, sulcatone, guaiacol, sulcatol, 2,4-dimethyl-3-hexanol, 2,4-dimethyl-2-hexanone, heptanal, nonanal, (Z)-3-hexenol, and 1-heptanol in both sexes, and by linalool, (E)-2-heptenal, 1-octen-3-ol, (E)-2-octenal, 3-octanol, (E)-2-octen-1-ol, α-phellandrene, and α-terpinene in males. The olfactory system of both sexes proved to be sensitive to changes in stimulus concentration and compound structure. CONCLUSIONS: this study demonstrates the capability of A. bungii males and females to detect and discriminate among a wide range of VOCs and provides a basis for further olfactometer and field trapping experiments aimed at identifying behaviorally-active compounds useful for the implementation of semiochemical-based control strategies for this pest.

Scanning Electron Microscopy of the Antennal Sensilla and Their Secretion Analysis in Adults of Aromia bungii (Faldermann, 1835) (Coleoptera, Cerambycidae).

BACKGROUND: It has been proved that chemical signals play an important role in mating location and reproductive behavior in cerambycids; moreover, they rely on contact chemoreception for mate recognition. METHODS: Adult antennae of Aromia bungii were observed using scanning electron microscopy and adult antennal secretions were collected and analyzed with gas chromatography-mass spectrometry. RESULTS: Twelve different types of sensilla were morphologically described on the antennae of A. bungii. At least six mechanoreceptors-one gustative, one putative chemo- or thermoreceptor, and three multiporous olfactory receptors-are present on the antennae of both sexes while a receptor-type of unclear function is limited to males. Secretions associated with sensilla basiconica were observed for the first time in a cerambycid species. CONCLUSIONS: Sensilla basiconica should play a role in odor perception detecting host tree volatiles and/or pheromones. Sensilla basiconica type 1 and 2 produce a viscous material accumulating on the antennal surface. Chemical analysis of adult antennal secretions highlighted marked differences between sexes. Some of the identified compounds have been previously reported as contact pheromone components of other cerambycid species. Our observations strongly suggest sensilla basiconica as the production sites of compounds involved in mate recognition.

Antennal olfactory responses of adult meadow spittlebug, Philaenus spumarius, to volatile organic compounds (VOCs).

The meadow spittlebug, Philaenus spumarius L. (Hemiptera, Aphrophoridae) is a commonly found vector of Xylella fastidiosa Wells et al. (1987) strain subspecies pauca associated with the "Olive Quick Decline Syndrome" in Italy. To contribute to the knowledge of the adult P. spumarius chemoreceptivity, electroantennographic (EAG) responses of both sexes to 50 volatile organic compounds (VOCs) including aliphatic aldehydes, alcohols, esters, and ketones, terpenoids, and aromatics were recorded. Measurable EAG responses were elicited by all compounds tested. In both sexes, octanal, 2-octanol, 2-decanone, (E)-2-hexenyl acetate, and vanillin elicited the strongest antennal amplitude within the chemical groups of aliphatic saturated aldehydes, aliphatic alcohols, aliphatic acetates and aromatics, respectively. Male and female EAG responses to sulcatol, (±)linalool, and sulcatone were higher than those to other terpenoinds. In both sexes, the weakest antennal stimulants were phenethyl alcohol and 2-pentanone. Sexual differences in the EAG amplitude were found only for four of test compounds suggesting a general similarity between males and females in antennal sensitivity. The olfactory system of both sexes proved to be sensitive to changes in stimulus concentration, carbon chain length, and compound structure. Compounds with short carbon chain length (C5-C6) elicited lower EAG amplitudes than compounds with higher carbon chain length (C9-C10) in all classes of aliphatic hydrocarbons with different functional groups. The elucidation of the sensitivity profile of P. spumarius to a variety of VOCs provides a basis for future identification of behaviorally-active compounds useful for developing semiochemical-based control strategies of this pest.