Behavioral Bioassays Research Articles

Gravid Periplaneta americana (Blattodea: Blattidae) Fails to Detect or Respond to the Presence of the Oothecal Parasitoid Aprostocetus hagenowii (Hymenoptera: Eulophidae).

Several families of parasitic Hymenoptera have evolved traits that allow them to exploit cockroach oothecae. Cockroaches may bury and conceal their oothecae to prevent parasitoid attack. However, these protective measures require additional investment by females. We hypothesized that gravid cockroaches would reduce parental care in the absence of oothecal parasitoids and increase care when parasitoids were detected. Behavior bioassays consisted of glass jars containing a gravid American cockroach, Periplaneta americana (L.) (Blattodea: Blattidae), expanded polystyrene (EPS), and a dog food pellet. A fruit fly (Drosophila melanogaster Meigen) (Diptera: Drosophilidae) or parasitoid Aprostocetus hagenowii (Ratzburg) (Hymenoptera: Eulophidae) was added for the fly and parasitoid treatments, respectively. There was no significant difference among treatments in the proportion of oothecae buried or in mean cover of oothecae with EPS particles. Cover had no effect on parasitism success or failure. Electroantennogram (EAG) assays using P. americana antennae were also conducted. The EAG responses to dead parasitoid stimuli (0.111-0.124 mV) were significantly (p < 0.05) greater than the negative control, but responses to living parasitoid stimuli (0.075-0.089 mV) were nonsignificant. These findings suggest that burial and concealment of oothecae is a general defensive behavior employed regardless of the presence or absence of a natural enemy. The results also indicate that gravid P. americana are unable to detect, and therefore, differentiate A. hagenowii from other insects and that A. hagenowii can successfully locate and parasitize oothecae completely concealed with EPS particles.

Powdery Mildew Fungus Oidium lycopersici Infected-Tomato Plants Attracts the Non-Vector Greenhouse Whitefly, Trialeurodes vaporariorum, but Seems Impair Their Development

The mechanism underlying the mediation of the behavior and fitness of non-vector herbivores by fungal pathogen-infected host plants is still unclear. In this study, we experimentally assessed the effects of tomato (Lycopersicon esculentum L.) plant infection status on non-vector herbivores using tomato powdery mildew disease fungus (Oidium lycopersici) and the greenhouse whitefly Trialeurodes vaporariorum as a model multitrophic plant-pathogen, non-vector herbivore interaction. In behavioral bioassays, powdery mildew-infected plants (PM) and plants damaged by combined attack of powdery mildew and whiteflies (PMWF) attracted significantly more whiteflies (32.6% and 58.1% for PM and PMWF, respectively) than control (Con) plants (16.5%) or plants damaged only by whiteflies (WF) (24.2%). Whiteflies feeding on PM or PMWF plants secreted almost twice as much honeydew as on the Con or WF plants. This indicated that whiteflies fed more on diseased plants than healthy plants. However, the preferences and increased feeding did not increase the fitness and immature growth of T. vaporariorum, but significantly prolonged their immature developmental durations, compared with those of whiteflies that developed on Con plants or WF plants. The results showed that powdery mildew infestation attracted more whiteflies but reduced their development and fitness.

Chemical Cues Used by the Weevil Curculio chinensis in Attacking the Host Oil Plant Camellia oleifera

The weevil Curculio chinensis Chevrolat (Coleoptera: Curculionidae) is a major cause of economic losses to growers of Camellia oleifera in China, as females lay their eggs in developing fruits and the hatching larvae feed on their seed, aborting fruit growth. Olfactory cues play a key role in the host location of this weevil. The present study focused on identifying volatiles from different parts of the host plant Ca. oleifera, namely, the leaves, fruit peel, and seeds, and testing the antennal and behavioral responses of adult Cu. chinensis to those same volatiles. Methods relied on gas chromatography, electroantennograms, and Y-tube bioassays. The results included a total of twenty-five volatiles emitted by the three plant parts, among which eight elicited antennal responses in Cu. chinensis adults of both sexes. The behavioral bioassays indicated that 3-hexenal, trans-2-hexen-1-ol, methyl salicylate, geraniol, and phenethyl alcohol were attractive to Cu. chinensis, while trans-2-hexenal and 2-ethyl-1-hexanol were repellent. Tests with different concentrations indicated that the behavioral response could be dose-dependent. Future studies should focus on field tests with blends of the attractant compounds in order to develop novel, improved control methods for field applications.

Repellent activity against Anopheles gambiae of the leaves of nesting trees in the Sebitoli chimpanzee community of Kibale National Park, Uganda.

BackgroundEvery evening, chimpanzees (Pan troglodytes) build a sleeping platform so called “nest” by intertwining branches of tree. Most of chimpanzees’ communities studied have a preference for tree species in which they nest. As female mosquitoes are feeding on the blood of their host at nighttime, chimpanzees may prevent being disturbed and bitten by mosquitoes by selecting tree species having properties to repel them.MethodsTo test the hypothesis that chimpanzees choose tree species for their aromatic properties, data related to 1,081 nesting trees built between 2017 and 2019 in the Sebitoli community of Kibale National Park (Uganda) were analysed. The 10 most used trees were compared to the 10 most common trees in the habitat that were not preferred for nesting. Leaves from the 20 trees species were collected and hydro-distillated to obtain essential oils and one of the by-products for behavioural bioassays against females of the African mosquito, Anopheles gambiae.ResultsSebitoli chimpanzees showed tree preferences: 10 species correspond to more than 80% of the nesting trees. Out of the essential oil obtained from the 10 nesting trees, 7 extracts for at least one concentration tested showed spatial repellency, 7 were irritant by contact and none were toxic. In the other hand, for the abundant trees in their habitat not used by chimpanzees, only 3 were repellent and 5 irritants.Discussion and conclusionThis study contributes to evidence that chimpanzees, to avoid annoying mosquitoes, may select their nesting trees according to their repellent properties (linked to chemical parameters), a potential inspiration for human health.Supplementary InformationThe online version contains supplementary material available at 10.1186/s12936-022-04291-7.

Plant volatile compound methyl benzoate is highly effective against Spodoptera frugiperda and safe to non-target organisms as an eco-friendly botanical-insecticide

Recent studies have indicated that the plant volatile methyl benzoate (MB) exhibits significant insecticidal bioactivity against several common insects. However, the potential environmental hazards of MB and its safety to non-target organisms is poorly understood. In the present study, these characteristics were investigated through laboratory experiments and field investigations. The results revealed that MB was highly toxic to the agricultural pest, fall armyworm Spodoptera frugiperda. Compared with the commercial pesticide lambda-cyhalothrin, the toxicities of MB against S. frugiperda larvae and adults were comparable and 3.41 times higher, respectively. Behavioral bioassays showed that the percentage repellency of MB to S. frugiperda larvae was 56.72 %, and MB induced 69.40 % oviposition deterrence rate in S. frugiperda female adults. Furthermore, in terms of median lethal concentration (LC50) and median lethal doses (LD50), MB exhibited non-toxic effects on non-target animals with 3-d LC50 of > 1 % to natural predators (Coccinella septempunctata and Harmonia axyridis), 3-d LD50 of 467.86 µg/bee to the bumblebee Bombus terrestris, 14-d LC50 of 971.09 mg/kg to the earthworm Eisenia fetida, and 4-d LC50 of 47.30 mg/L to the zebrafish Brachydanio rerio. The accumulation of MB in the soil and earthworms was found to be extremely limited. Our comparative study clearly demonstrated that MB is effective as a selective botanical pesticide against S. frugiperda and it is safe to use in the tested environment, with no toxic effects on non-target animals and natural predators.

Same sensitivity with shorter exposure: behavior as an appropriate parameter to assess metal toxicity

The exposure of animals to toxicants may cause a depletion in the energy uptake, which compromises reproduction and growth. Although both parameters are ecologically relevant, they usually need long-term bioassays. This is a handicap for the availability of toxicological data for environmental risk assessment. Short-term bioassays conducted with environmental concentrations, and using relevant ecological parameters sensitive to short-term exposures, such as behavior, could be a good alternative. Therefore, to include this parameter in the risk assessment procedures, it is relevant the comparison of its sensitivity with that of growth and reproduction bioassays. The study aim was the assessment of differences between endpoints based on mortality, behaviour, reproduction, and growth for the toxicity of metals on aquatic animals. We used the ECOTOX database to gather data to construct chemical toxicity distribution (CTD) curves. The mean concentrations, the mean exposure time, and the ratio between the mean concentration and the exposure time were compared among endpoints. Our results showed that behavioral, growth, and reproduction bioassays presented similar sensitivity. The shortest exposure was found in behavioral and reproduction bioassays. In general, the amount of toxicant used per time was lower in growth and reproduction bioassays than in behavioral and mortality bioassays. We can conclude that, for metal toxicity, behavioral bioassays are less time-consuming than growth bioassays. As the sensitivity of behavior was similar to that of growth and reproduction, this endpoint could be a better alternative to longer bioassays.

Identification of Putative Neuropeptides That Alter the Behaviour of Schistosoma mansoni Cercariae.

Simple SummarySchistosoma mansoni is a common etiological agent of human schistosomiasis, one of the deadliest neglected tropical diseases in the developing world. Schistosoma neuropeptides have been considered a possible avenue of targeted biocontrol. We aimed to identify species-specific putative neuropeptides derived from the mammalian-infective stage, cercariae, which may function as biocontrol targets. A total of 11 neuropeptide precursors were identified for the first time in cercariae, all of which were highly specific to Schistosoma and many of which were highly expressed in the cercarial stage. We tested nine putative neuropeptides derived from these precursor proteins, three of which induced a significant increase in cercariae stopping, turning and passive behaviour over periods of both 1 min and 360 min post-exposure. These observations implicate the tested neuropeptides as involved in regulating cercarial behaviour. The characterisation of these putative neuropeptides may facilitate the innovation of biocontrols to prevent infection.Elucidating the infectivity of Schistosoma mansoni, one of the main etiological agents of human schistosomiasis, requires an improved understanding of the behavioural mechanisms of cercariae, the non-feeding mammalian infective stage. This study investigated the presence and effect of cercariae-derived putative neuropeptides on cercarial behaviour when applied externally. Cercariae were peptidomically analysed and 11 neuropeptide precursor proteins, all of which were specific to the Schistosoma genus and most of which highly expressed in the cercarial stage, were identified in cercariae for the first time. Protein–protein interaction analysis predicted the interaction of various neuropeptide precursors (e.g., Sm-npp-30, Sm-npp-33, Sm-npp-35) with cercarial structural proteins (e.g., myosin heavy chain and titin). In total, nine putative neuropeptides, selected based on their high hydrophobicity and small size (~1 kilodalton), were tested on cercariae (3 mg/mL) in acute exposure (1 min) and prolonged exposure (360 min) behavioural bioassays. The peptides AAYMDLPW-NH2, NRKIDQSFYSYY-NH2, FLLALPSP-OH, and NYLWDTRL-NH2 stimulated acute increases in cercarial spinning, stopping, and directional change during active states. However, only NRKIDQSFYSYY-NH2 caused the same behavioural changes at a lower concentration (0.1 mg/mL). After prolonged exposure, AAYMDLPW-NH2 and NYLWDTRL-NH2 caused increasing passive behaviour and NRKIDQSFYSYY-NH2 caused increasing body-first and head-pulling movements. These findings characterise behaviour-altering novel putative neuropeptides, which may inform future biocontrol innovations to prevent human schistosomiasis.

Links Between Feeding Preferences and Electroantennogram Response Profiles in Dung Beetles: The Importance of Dung Odor Bouquets

The detection of dung odors is a crucial step in the food-searching behavior of dung beetles (Coleoptera: Scarabaeoidea). Yet, whether certain compounds characteristic of a given dung type contribute to a ‘choosy generalism’ behavior proposed for this taxonomic group is unknown. To address this, we analyzed the chemical composition of three types of dung (cow, horse, and rabbit) and conducted behavioral and electroantennogram (EAG) bioassays on 15 species of dung beetles using 19 volatile organic compounds representing the three dung samples. Chemical analyses revealed substantial qualitative and quantitative differences among dung types. When offered these food options in an olfactometer, 14 species exhibited a feeding preference. Surprisingly, all 19 compounds used in the EAG assays elicited antennal responses, with species displaying different olfactory profiles. The relationship between behavioral preferences and electrophysiological profiles highlighted that species with different food preferences had differences in antennal responses. Moreover, a specific set of EAG-active compounds (nonanal, sabinene, acetophenone, ρ-cresol, 2-heptanone, 1H-indole, and 6-methyl-5-hepten-2-one) were the strongest drivers in the distinct sensory profiles of the trophic preference groups. Our results point to the importance of the whole bouquet of dung-emanating compounds in driving food-searching behavior, but specific volatiles could aid in determining highly marked trophic preferences in certain species.

Sensory-Motor Perturbations in Larval Zebrafish (Danio rerio) Induced by Exposure to Low Levels of Neuroactive Micropollutants during Development.

Due to increasing numbers of anthropogenic chemicals with unknown neurotoxic properties, there is an increasing need for a paradigm shift toward rapid and higher throughput behavioral bioassays. In this work, we demonstrate application of a purpose-built high throughput multidimensional behavioral test battery on larval stages of Danio rerio (zebrafish) at 5 days post fertilization (dpf). The automated battery comprised of the established spontaneous swimming (SS), simulated predator response (SPR), larval photomotor response (LPR) assays as well as a new thermotaxis (TX) assay. We applied the novel system to characterize environmentally relevant concentrations of emerging pharmaceutical micropollutants including anticonvulsants (gabapentin: 400 ng/L; carbamazepine: 3000 ng/L), inflammatory drugs (ibuprofen: 9800 ng/L), and antidepressants (fluoxetine: 300 ng/L; venlafaxine: 2200 ng/L). The successful integration of the thermal preference assay into a multidimensional behavioral test battery provided means to reveal ibuprofen-induced perturbations of thermal preference behaviors upon exposure during embryogenesis. Moreover, we discovered that photomotor responses in larval stages of fish are also altered by the as yet understudied anticonvulsant gabapentin. Collectively our results demonstrate the utility of high-throughput multidimensional behavioral ecotoxicity test batteries in prioritizing emerging risks associated with neuroactive drugs that can perturb neurodevelopment. Moreover, we showcase the added value of thermotaxis bioassays for preliminary screening of emerging contaminants.

Trophic specialization of a newly described spider ant symbiont, Myrmecicultor chihuahuensis (Araneae: Myrmecicultoridae)

The spider Myrmecicultor chihuahuensis Ramírez, Grismado & Ubick (Myrmecicultoridae) was described in 2019 and hypothesized to be a myrmecophile, living inside the nests of Novomessor (Myrmicinae) and perhaps also Pogonomyrmex (Myrmicinae) ants. To test the hypothesis that M. chihuahuensis are chemical mimics of their host ants, we carried out behavioral bioassays to observe interactions between the spiders and the host ants. We compared the cuticular hydrocarbon (CHC) profiles of the spiders and the ants. We discovered that this new species of spider is a myrmecophage, displaying hunting strategies similar to other myrmecophagous spiders, particularly those in the genus Zodarion Walckenaer, 1826 (Zodariidae). The CHC profile of M. chihuahuensis did not match the profile of the ants with which it associates, Novomessor albisetosus (Mayr), but ants and spider shared several compounds, potentially involved in species recognition.

An odorant binding protein mediates Bactrocera dorsalis olfactory sensitivity to host plant volatiles and male attractant compounds

Odorant-binding proteins (OBPs) are believed to play critical roles in host-seeking behavior. However, little attention was paid to its different functions in male and female. The antenna-specific OBP gene from Bactrocera dorsalis, BdorOBP13, was cloned and its expression profile was examined. The results showed that BdorOBP13 was exclusively expressed in male and female adults, which exhibited a high transcript level in antennae. After injection of BdorOBP13 dsRNA, its transcript level in males and females decreased significantly. Electrophysiological responses of RNAi-injected flies to, methyl eugenol (male attractant) and γ-octalactone (female attractant) decreased significantly. However, no significant changes in the electrophysiological response were observed in RNAi-injected flies to benzothiazole, (+),dipentene, and ethyl tiglate. The behavioral bioassay showed that males treated with RNAi significantly reduced their preference to methyl eugenol, while RNAi-injected females showed a significantly lower preference to γ-octalactone, suggesting that BdorOBP13 may have different functions between males and females: it may be involved in the detection of methyl eugenol in males but is involved in the detection of γ-octalactone in females. These findings improve our understanding of insect OBPs and their roles in insect chemosensation, which may provide us with new molecular targets in the management of B. dorsalis.

Electrophysiological and behavioural responses of Stegobium paniceum to volatile compounds from Chinese medicinal plant materials.

BACKGROUND Stegobium paniceum (Coleoptera, Anobiidae) is an important pest of stored products causing severe damage to dried Chinese medicinal plant materials (CMPMs). Plant volatiles play an important role in host‐searching of insects. The olfactory responses of S. paniceum to the most abundant volatile components of some drugstore attractant CMPMs such as Panax notoginseng, Angelica sinensis, Gastrodia elata and Peucedanum praeruptorum, namely falcarinol, 3‐n‐butylphthalide, p‐cresol and β‐pinene, respectively, were studied by electroantennography (EAG) and behavioural bioassays in six‐ and four‐arm olfactometers.RESULTSEAG recordings showed that male and female antennae are able to perceive the test compounds in a wide range of concentrations and in a dose‐dependent manner. Moreover, for each dose of different compounds tested, no significant differences were found between the mean male and female EAG responses. In six‐arm olfactometer bioassays, S. paniceum exhibited positive responses to falcarinol, 3‐n‐butylphthalide, p‐cresol and β‐pinene at doses of 1, 10, 100, 500 and 1000 μg. The most attractive dose was 500 μg for falcarinol, 100 μg for 3‐n‐butylphthalide, 500 μg for p‐cresol and 1000 μg for β‐pinene. Olfactory preferences of S. paniceum, based on comparison of these four compounds at their optimally attractive concentrations in a four‐arm olfactometer, were 3‐n‐butylphthalide > p‐cresol > falcarinol > β‐pinene.CONCLUSIONThe results indicated that the four volatiles of CMPMs are perceived by the peripheral olfactory system of S. paniceum adults and are able to individually elicit a positive chemotaxis in S. paniceum adults confirming the role of chemical cues in host‐plant detection and selection of this pest. Further field studies are needed to evaluate the potential of the attractive compounds identified in this study, particularly 3‐n‐butylphthalide, to be applied as a novel monitoring and control tool against this storage‐beetle pest. © 2022 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Identification and functional analysis of a chemosensory protein from Bactrocera minax (Diptera: Tephritidae).

Olfaction has an indispensable role in insect behavior, enabling location of suitable host plants and oviposition sites, finding mates and evasion of natural enemies. Chemosensory proteins (CSPs) function to screen external odorants and transport them to olfactory receptor neurons, thereby increasing the sensitivity of the olfactory system. At present, CSP genes have been identified in many insect species, but there are relatively few studies on the function of CSP, especially in Tephritidae. In this study, we sequenced and analyzed 12 transcriptomes of Bactrocera minax and identified five CSP genes. The results of polymerase chain reactions with reverse transcription showed that BminCSP3 was highly expressed only in antennae. Results from competitive binding experiments showed that BminCSP3 has good binding ability to citral compared with 23 other volatile organic compounds. The docking model with citral showed hydrogen bond formation with residues (ARG97); however, no hydrogen bonds were formed in the docking of five other ligands (furfuryl alcohol, linalool, cis-3-hexenyl acetate, (R)-(+)-limonene and (+)-carvone). Electroantennogram (EAG) analyses revealed that citral was active in B. minax at the antennal level, and the EAG response value of female adults was significantly higher than that of male adults. Furthermore, the results of behavioral bioassays showed that females were significantly attracted to citral. Our results suggest that BminCSP3 plays an important role in the recognition of citral by B. minax adults. © 2022 Society of Chemical Industry.

In a behavioural bioassay, effluents from cigarette butts, cured tobacco and tree leaves differentially affect resting site selection in the land snail Cornu aspersum

ABSTRACT Cigarette butts (CBs) contain a plethora of chemicals, including many that are toxic. Although numerous studies have demonstrated the toxicity of CBs to aquatic organisms, there is less evidence that terrestrial organisms are severely impacted. Because CBs are commonly discarded on the ground, ground-dwelling organisms such as land snails may be especially affected. Many land snails are generalist detritivores/herbivores and ingest a variety of plant secondary compounds as they feed. This evolutionary exposure may render CBs (made principally of cured tobacco leaves) less toxic to these land snails than CBs are to less exposed aquatic animals. We investigated this possibility of reduced effects using a new behavioural assay to test the choice of ‘flavoured’ vertical resting sites in the land snail Cornu aspersum, which commonly rests on upright surfaces. In four experiments, regions of container walls were coated with different concentrations of CB and cured tobacco effluents, and effluents from three tree species and cured tobacco. Snails avoided high CB and tobacco effluent concentrations, which is consistent with toxicity. However, snails preferentially rested on dilute concentrations of both CBs and tobacco. Preference among tree leaf effluents was less evident, with a trend towards snails preferring the more readily eaten maple than the less readily eaten oak. Selection of the preferred tobacco concentration did not differ from that of tree leaves, indicating that C. aspersum was not repelled by dilute tobacco effluent. These results indicate that compounds leaching from discarded CBs may have little effect on snails and perhaps other soil organisms under environmentally realistic conditions.

Functional Differentiation of Floral Color and Scent in Gall Midge Pollination: A Study of a Schisandraceae Plant.

Gall midges are among the most host-specific insects. Their interactions with plants likely date back to the Cretaceous period. Plants from at least seven families are involved in gall midge pollination; however, little is known about the pollination signals of gall midges. In this study, we used a Resseliella–Schisandra model to investigate the roles of floral scent and color in attracting gall midges. Field observations, behavioral bioassays via Y-tubes, and “flight box” experiments were performed. The results demonstrated that gall midges may be attracted by both floral scent and color and that two flower signals are more effective in promoting insect flower-landing than either alone. In the field, gall midges visited male flowers effectively at night but almost always visited female flowers during the day. Thus, during the Resseliella–Schisandra interactions, female flowers predominantly employed visual cues over scent to attract midges during the day; in contrast, olfactory cues were more functional for male flowers to export pollen in the dark. In this study, we first identified the roles of floral color and the functional differentiation of visual and olfactory cues during gall midge pollination.

Larval salmon lice Lepeophtheirus salmonis exhibit behavioral responses to conspecific pre-adult and adult cues.

In the larval stage of the parasitic copepod Lepeophtheirus salmonis, the free-living copepodid must locate and settle on a salmonid host. Chemosensory mechanisms play a role in determining whether a potential host is suitable for attachment, yet the full suite of chemical cues and resulting behavioral mechanisms used are unknown. After maturing, pre-adult female and adult male salmon lice aggregate upon salmonid hosts for reproduction. Copepodid salmon lice have been observed preferentially infesting hosts that harbor conspecific adults. The aim of this study was to investigate the possibility that salmon lice copepodids perceive and respond to cues from pre-adult female, adult-male, and/or gravid female salmon lice. Behavioral bioassays were conducted in vitro with copepodids exposed to water conditioned with 3 stages of conspecific lice (pre-adult female, adult male, or gravid female), and seawater conditioned with Atlantic salmon Salmo salar Linnaeus, 1758. Experiments demonstrated that copepodids exposed to water conditioned with the salmon host, pre-adult female, or adult male salmon lice significantly altered their behavior, whereas salmon lice exposed to water conditioned with gravid females did not. These results are potentially valuable in the development of novel methods for mitigation of L. salmonis in the salmon aquaculture industry.

Control of diamondback moth with Lippia gracilis essential oil

Abstract The objective of this work was to evaluate the potential of the essential oils of the LGRA-106 and LGR A-108 Lip pia g racili s genot y pes for the control of diamondback moth (Plutella xylostella). The lethal concentrations (LCs) were estimated by two routes of action (residual and spraying), using oil concentrations ranging from 0.5 to 3.0% v v−1, diluted in Tween 80 (1.5%). To determine the effect of the LC to 50% (LC50) on the development of P. xylostella, two compounds of the L. gracilis genotypes, thymol, and carvacrol, were sprayed on the insects. The repellency of the LC50 was evaluated by residual action, in a free-choice behavioral bioassay. The LGRA-106 genotype showed a greater toxicity via residual action (LC50 = 8.82 mg mL−1), as well as a higher repellency index. LGRA-108 was more toxic via spraying (LC50= 9.64 mg mL−1). Larval development and viability were reduced in approximately 50% with LGRA-106 or thymol and up to 70% with LGRA-108 or carvacrol, which caused mortality from 1.70 to 1.97 days after spraying. The oils of the LGRA-106 and LGRA-108 genotypes of L. gracilis have insecticidal activity in the control of P. xylostella.

Chemical Composition of Essential Oils from Leaves and Fruits of Juniperus foetidissima and Their Attractancy and Toxicity to Two Economically Important Tephritid Fruit Fly Species, Ceratitis capitata and Anastrepha suspensa.

The present study analyzed the chemical composition of Juniperus foetidissima Willd. essential oils (EOs) and evaluated their attractancy and toxicity to two agriculturally important tephritid fruit flies. The composition of hydrodistilled EOs obtained from leaves (JFLEO) and fruits (JFFEO) of J. foetidissima was analyzed by GC–FID and GC–MS. The main compounds were α-pinene (45%) and cedrol (18%) in the JFLEO and α-pinene (42%), α-thujone (12%), and β-thujone (25%) in the JFFEO. In behavioral bioassays of the male Mediterranean fruit fly, Ceratitis capitata (Wiedemann), both JFLEO and JFFEO showed strong attraction comparable to that observed with two positive controls, Melaleuca alternifolia and Tetradenia riparia EOs. In topical bioassays of the female Caribbean fruit fly, Anastrepha suspensa (Loew), the toxicity of JFFEO was two-fold higher than that of JFLEO, with the LD50 values being 10.46 and 22.07 µg/µL, respectively. This could be due to differences in chemical components between JFLEO and JFFEO. The JFFEO was dominated by 48% monoterpene hydrocarbons (MH) and 46% oxygenated monoterpenes (OM), while JFLEO consisted of 57% MH, 18% OM, and 20% oxygenated sesquiterpenes (OS). This is the first study to evaluate the attractancy and toxicity of J. foetidissima EOs to tephritid fruit flies. Our results indicate that JFFEO has the potential for application to the management of pest tephritid species, and further investigation is warranted.

Herbivore-induced volatiles influence moth preference by increasing the β-Ocimene emission of neighbouring tea plants.

Herbivore-induced plant volatiles prime neighbouring plants to respond more strongly to subsequent attacks. However, the key volatiles that trigger this state and their priming mechanisms remain largely unknown. The tea geometrid Ectropis obliqua is one of the most devastating leaf-feeding pests of tea plants. Here, plant-plant communication experiments demonstrated that volatiles emitted from tea plants infested by E. obliqua larvae triggered neighbouring plants to release volatiles that repel E. obliqua adult, especially mated females. Volatile analyses revealed that the quantity of eight volatiles increased dramatically when plants were exposed to volatiles emitted by infested tea plants, including (Z)-3-hexenol, linalool, α-farnesene, β-Ocimene and (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT). The results of behavioural bioassays demonstrated that β-Ocimene strongly repelled mated E. obliqua females. Individual volatile compound exposure experiments revealed that (Z)-3-hexenol, linalool, α-farnesene and DMNT triggered the emission of β-Ocimene from tea plants. Chemical inhibition experiments demonstrated that the emission of β-Ocimene induced by (Z)-3-hexenol, linalool, α-farnesene and DMNT were dependent on Ca2+ and JA signalling. These findings help us to understand how E. obliqua moths respond to volatiles emitted from tea plants and provide new insight into volatile-mediated plant-plant interactions. They have potential significance for the development of novel insect and pest control strategies in crops.

Behavior Matters-Future Need for Insect Studies on Odor-Mediated Host Plant Recognition with the Aim of Making Use of Allelochemicals for Plant Protection.

Allelochemicals, chemical cues that, among other things, mediate insect-plant interactions, such as host plant recognition, have attracted notable interest as tools for ecological control of pest insects. Advances have recently been made in methods for sampling and analyzing volatile compounds and technology for tracking insects in their natural habitat. However, progress in odor-mediated behavioral bioassays of insects has been relatively slow. This perspective highlights this odor-mediated insect behavior, particularly in a natural setting and considering the whole behavioral sequence involved in the host location, which is the key to understanding the mechanisms underlying host plant recognition. There is thus a need to focus on elaborate behavioral bioassays in future studies, particularly if the goal is to use allelochemicals in pest control. Future directions for research are discussed.