Multiple factors influenced the aggregation behavior of adult Eucryptorrhynchus scrobicuatus (Coleoptera: Curculionidae).

Eucryptorrhynchus scrobiculatus (Motschulsky) (Coleoptera: Curculionidae) is a notorious pest of Ailanthus altissima (Mill.) Swingle (Sapindales: Simaroubaceae). E. scrobiculatus adults typically aggregate under leaves and in soil crevices at the base of A. altissima in the field. We hypothesize that the environmental factors and conspecific signals determine their aggregation behavior. To test this, we investigated adult numbers in light-exposed and shaded areas of the sample trees and conducted experiments in both field and lab settings. Results revealed that (i) greater adult distribution in shaded areas; (ii) significant influence of temperature and illumination on aggregation tendency in the field; (iii) no gender-based difference in aggregation degree and maximum aggregation between light and dark; (iv) the host plant triggering the aggregation tendency, negatively affected in the absence; (v) the aggregation tendency of E. scrobiculatus weakened with the temperature gradually changing to ordinary temperature; and (vi) mutual attraction and chemical attraction between males and females. Thus, the aggregation behavior was influenced by factors including temperature, light intensity, host plant, and conspecific signals, but light's role was not obvious in the lab.

Bumblebees attend to both the properties of the string and the target in string-pulling tasks, but prioritize the features of the string.

Previous studies have demonstrated that associative learning and experience play important roles in the string-pulling of bumblebees (Bombus terrestris). However, the features of the target (artificial flower with sugar reward) and the string that bees learn in such tasks remain unknown. This study aimed to explore the specific aspects of the string-flower arrangement that bumblebees learn and how they prioritize these features. We show that bumblebees trained with string-pulling are sensitive to the flower stimuli; they exhibit a preference for pulling strings connected to flowers over strings that are not attached to a target. Additionally, they chose to pull strings attached to flowers of the same color and shape as experienced during training. The string feature also plays a crucial role for bumblebees when the flower features are identical. Furthermore, bees prioritized the features of the strings rather than the flowers when both cues were in conflict. Our results show that bumblebees solve string-pulling tasks by acquiring knowledge about the characteristics of both targets and strings, and contribute to a deeper understanding of the cognitive processes employed by bees when tackling non-natural skills.

Object use in insects.

Insects are the most diverse group of organisms in the animal kingdom, and some species exhibit complex social behaviors. Although research on insect object use is still in its early stages, insects have already been shown to display rich object-use behaviors. This review focuses on patterns and behavioral flexibility in insect object-use behavior, and the role of cultural evolution in the development of object-use behaviors. Object use in insects is not widespread but has been documented in a diverse set of taxa. Some insects can use objects flexibly and display various object-use patterns. Like mammals and birds, insects use objects in diverse activities, including foraging, predator defense, courtship, and play. Intelligence, pre-existing manipulative behaviors, and anatomical structure affect innovations in object use. In addition, learning and imitation are the main mechanisms underlying the spread of object-use behaviors within populations. Given that insects are one of the major animal groups engaging in object use, studies of insect object use could provide general insights into object use in the animal kingdom.

Observations on neurophysiological pattern and behavioural traits as death-feigning mechanism in Eucryptorrhynchus scrobiculatus (Coleoptera: Curculionidae).

In adaptation to surrounding environmental stimuli, most insects exhibit defense behaviour (death feigning) to improve survival rates in the wild. However, the underlying mechanism of death feigning remains largely unknown. Here, we tested the neurophysiological pattern and behavioural traits of the death-feigning mechanism in the forestry pest Eucryptorrhynchus scrobiculatus. Using neuroanatomy, LC-MS/MS target metabolomics detection technology and qRT-PCR, we investigated the effects of neurochemicals and metabolic pathways in experimental weevils. Excision and drug tests were conducted to verify the key regulatory body parts involved in regulating the central nervous system in death feigning. Our results reconstructed the death-feigning mechanism of E. scrobiculatus: when the effective stimuli point of arousal weevils received mechanical stimulation, the thoracoabdominal ganglion transmitted signals into the brain through the ventral nerve cord, and then the brain regulated dopamine (DA) and serotonin (5-HT) metabolic pathways, reducing the expression of dopamine (dar2) and octopamine (oar1, oab2) receptor genes, finally inducing death feigning. Our study suggests that the variation of neurotransmitters in the brain is an important indicator of the physiological response of death feigning, and the results provide ecological and theoretical information for future investigations to reveal key behaviour and target genes for pest control.

Comparison of Fine Structure of the Compound Eyes in Eucryptorrhynchus scrobiculatus and Eucryptorrhynchus brandti Adults.

Eucryptorrhynchus scrobiculatus and E. brandti are the main borers of Ailanthus altissima, causing serious economic and ecological losses. The external morphology and internal ultrastructure of the compound eyes of two related weevils were investigated with light microscopy, scanning electron microscopy, and transmission electron microscopy. E. scrobiculatus and E. brandti possess a pair of reniform apposition compound eyes and contain about 550 ommatidia per eye. The interommatidial angle of E. scrobiculatus and E. brandti are 7.08 ± 0.31° and 4.84 ± 0.49°, respectively. The corneal thickness, rhabdom length, and ommatidium length of E. scrobiculatus are significantly greater than those of E. brandti. Under light-adapted conditions, the pigment granules are mainly distributed at the junction of the cone and the rhabdom, and the diameter and the cross-sectional area of the middle end of the rhabdom is increased in the two weevil species. Under dark-adapted conditions, the pigment granules shift longitudinally and are evenly distributed on both sides of the cone and the rhabdom, and the diameter and cross-sectional area of the middle end of the rhabdom are decreased. The discrepancy in visual structure is beneficial for adaptation to niche differentiation of the two related species. The present results suggest that the two weevils possess different visual organ structures to perceive visual information in the external environment.

Effects of supplemental nutrients on ovarian development and oviposition in Conogethes punctiferalis (Lepidoptera: Crambidae).

Conogethes punctiferalis is a serious pest in China affecting a wide variety of field crops, fruits, and forest trees. Many insects require supplemental nutrients after emergence to compensate for insufficient nutrients at the larval stage. In this study, to better understand the determinants of C. punctiferalis survival and reproduction, the impact of supplemental nutrition was examined. In particular, the effects of 11 treatments (5% sucrose, 10% sucrose, 15% sucrose, 5% sophora honey, 10% sophora honey, 15% sophora honey, 5% wild honey, 10% wild honey, 15% wild honey, distilled water, and blank control) on adult longevity and oviposition were evaluated, with detailed morphological analyses of ovarian development in the 10% sucrose, distilled water, and blank control groups. Conogethes punctiferalis required supplementary nutrition after emergence. Supplementary nutrition improved fecundity (number of eggs laid) and longevity, and 10% sucrose water had the greatest effects. The preoviposition period and oviposition period of C. punctiferalis females were longer, and the numbers of eggs in female ovarian tubes and eggs laid by females were higher in the 10% sucrose water group than in other groups. Females supplemented with distilled water laid a small number of eggs. Without nutrient supplementation, females did not lay eggs. In conclusion, supplemental nutrition was beneficial for ovarian development in female moths, prolonging the oviposition period and lifespan, and was an important factor affecting population dynamics. These results lay a foundation for further analyses of the nutritional requirements for C. punctiferalis in the field and provide a reference for indoor population feeding.

The Influencing Factors of Aggregation Behavior of Tree-of-Heaven Trunk Weevil, Eucryptorrhynchus brandti (Harold) (Coleoptera: Curculionidae).

The tree-of-heaven trunk weevil, Eucryptorrhynchus brandti (Harold) (Coleoptera: Curculionidae), is one of the most harmful pests that damage the tree of heaven, Ailanthus altissima Swingle (Sapindales: Simaroubaceae). Aggregation behavior tests of E. brandti adults were conducted in laboratory conditions. The effects of temperature and light on the aggregation behavior of adults were tested, and the effect of sex and host was conducted with binomial choice experiments. The results showed that (1) the adults aggregate in both light and dark environments but preferred the dark environment, (2) temperature can drive the aggregation of E. brandti adults, (3) host plants could trigger E. brandti adults' aggregation behavior, which is probably related to phytochemicals and insect feeding and localization, (4) there was mutual attraction of males and females and chemical attraction of crude intestinal extracts of males and females, and (5) aggregation behavior of E. brandti adults may also be related to the mediating of physical signals in insects. In this study, aggregation behavior can help us understand conspecific interactions and discover some strategies for effective control.

Morphologic Characters of the Rostrum in Two Weevils, Eucryptorrhynchus scrobiculatus Motschulsky and E. brandti Harold (Coleoptera: Curculionidae: Cryptorrhychinae).

(1) Eucryptorrhynchus scrobiculatus and E. brandti (Coleoptera: Curculionidae: Cryptorrhychinae) are both pests of Ailanthus altissima, found in China. During ovipositing, gravid females of the two weevils need to excavate a cavity in the oviposition substrate with their rostrum, while their oviposition sites are different. (2) In this study, to explore the boring mechanism of E. scrobiculatus and E. brandti during ovipositing, the morphologic characters of the rostra of two weevils were studied in detail by scanning electron microscopy and micro-CT. (3) Their rostra appear similar, but the rostrum surface of E. scrobiculatus is rougher than that of E. brandti; their fine structures of rostrum and sensilla distribution are similar, but the sensilla twig basiconica 3 is distributed at the apex of labial palpus in E. brandti females, while not at the apex of labial palpus in E. scrobiculatus females; their rostra are hollow and their cuticle thickness is constantly changing, but the proportion of the whole rostrum tube cuticle in E. scrobiculatus is significantly larger than that of E. brandti. The above structural differences make E. scrobiculatus more conducive to oviposition in the soil and E. brandti more conducive to oviposition in the trunk of A. altissima. (4) Overall, this study not only plays an important role in exploring the excavating mechanism during the oviposition of the two weevils, but also provides new insights into the coexistence of two weevil species on the same host A. altissima.

Effects of feeding on different parts of Ailanthus altissima on the intestinal microbiota of Eucryptorrhynchus scrobiculatus and Eucryptorrhynchus brandti (Coleoptera: Curculionidae).

Eucryptorrhynchus brandti and Eucryptorrhynchus scrobiculatus (Coleoptera: Curculionidae) are two monophagous weevil pests that feed on Ailanthus altissima (Mill.) Swingle but differ in their diet niche. In the field, adults of E. brandti prefer to feed on the trunk of A. altissima, whereas adults of E. scrobiculatus prefer to feed on the tender parts. We conducted Illumina sequencing of 16S rRNA to examine changes in bacterial diversity in the adults of these two weevil species after they fed on different parts of A. altissima (trunk, 2-3-year-old branches, annual branches, and petioles). Proteobacteria, Tenericutes, and Firmicutes were the dominant phyla in E. brandti (relative abundance was 50.64, 41.56, and 5.63%, respectively) and E. scrobiculatus (relative abundance was 78.63, 11.91, and 7.41%, respectively). At the genus level, Spiroplasma, endosymbionts2, Unclassified Enterobacteriaceae, and Lactococcus were dominant in E. brandti, and Unclassified Enterobacteriaceae, Wolbachia and Spiroplasma, and endosymbionts2 were dominant in E. scrobiculatus. Linear discriminant analysis effect size analysis revealed microbial biomarkers in the different treatment group of adults of both weevil species. Adults of E. brandti may require the trunk, and adults of E. scrobiculatus may require the petioles and annual branches to maintain the high diversity of their gut microbes. The results of this study indicate that feeding on different parts of A. altissima affects the composition and function of the microbes of E. brandti and the microbial composition of E. scrobiculatus. Variation in the abundance of Wolbachia and Spiroplasma in E. brandti and E. scrobiculatus is associated with dietary niche changes, and this might explain the evolution of reproductive isolation between these two sibling weevil species.

Population differentiation and intraspecific genetic admixture in two Eucryptorrhynchus weevils (Coleoptera: Curculionidae) across northern China.

Increasing damage of pests in agriculture and forestry can arise both as a consequence of changes in local species and through the introduction of alien species. In this study, we used population genetics approaches to examine population processes of two pests of the tree-of-heaven trunk weevil (TTW), Eucryptorrhynchus brandti (Harold) and the tree-of-heaven root weevil (TRW), E. scrobiculatus (Motschulsky) on the tree-of-heaven across their native range of China. We analyzed the population genetics of the two weevils based on ten highly polymorphic microsatellite markers. Population genetic diversity analysis showed strong population differentiation among populations of each species, with F ST ranges from 0.0197 to 0.6650 and from -0.0724 to 0.6845, respectively. Populations from the same geographic areas can be divided into different genetic clusters, and the same genetic cluster contained populations from different geographic populations, pointing to dispersal of the weevils possibly being human-mediated. Redundancy analysis showed that the independent effects of environment and geography could account for 93.94% and 29.70% of the explained genetic variance in TTW, and 41.90% and 55.73% of the explained genetic variance in TRW, respectively, indicating possible impacts of local climates on population genetic differentiation. Our study helps to uncover population genetic processes of these local pest species with relevance to control methods.

Climate Change Impacts on the Potential Distribution of Apocheima cinerarius (Erschoff) (Lepidoptera: Geometridae).

Among the impacts of ongoing and projected climate change are shifts in the distribution and severity of insect pests. Projecting those impacts is necessary to ensure effective pest management in the future. Apocheima cinerarius (Erschoff) (Lepidoptera: Geometridae) is an important polyphagous forest pest in China where causes huge economic and ecological losses in 20 provinces. Under historical climatic conditions, the suitable areas for A. cinerarius in China are mainly in the northern temperate zone (30-50° N) and the southern temperate zone (20-60° S). Using the CLIMEX model, the potential distribution of the pest in China and globally, both historically and under climate change, were estimated. Suitable habitats for A. cinerarius occur in parts of all continents. With climate change, its potential distribution extends northward in China and generally elsewhere in the northern hemisphere, although effects vary depending on latitude. In other areas of the world, some habitats become less suitable for the species. Based on the simulated growth index in CLIMEX, the onset of A. cinerarius would be earlier under climate change in some of its potential range, including Spain and Korea. Measures should anticipate the need for prevention and control of A. cinerarius in its potential extended range in China and globally.

A new management tactic for Eucryptorrhynchus scrobiculatus (Coleoptera: Curculionidae) based on factors influencing the weevil population density.

BACKGROUND: The tree of heaven root weevil, Eucryptorrhynchus scrobiculatus (TRW), is one of the most damaging pests of Ailanthus altissima in China. A limited number of studies have examined the effects of different variables on the population density of TRW adults. We investigated the relative impacts of the diameter at breast height of host trees (DBH), number of tree species in plots (NTS), weed coverage rate of the host tree base (WCR), plot area (PA), distance from plots to the nearest farmland (DNF), nearest human community (DNHC), nearest irrigation canal (DNIC), and nearest road (DNR) on TRW population density. We then evaluated the effect of removing weeds at the base of the tree trunk (RW) on TRW population density. RESULTS: We found that DBH, WCR, PA, and DNHC significantly affected the population density of TRW adults, whereas NTS, DNF, DNIC, and DNR did not. Adult TRW density was positively correlated with DBH, WCR, and PA but was negatively correlated with DNHC. Further field trials showed that RW treatments significantly reduced the density of adult TRW (an average annual reduction of 51.7% compared to untreated trees). CONCLUSION: This information on the factors affecting the population density of TRW promoted the development of new TRW management strategies. Our results demonstrate that RW is an effective method for reducing the population density of TRW adults. Therefore, this new physical method using RW could be recommended as an alternative for TRW management in the framework of integrated pest management. © 2021 Society of Chemical Industry.

Reproductive Dormancy in Overwintering Adult Eucryptorrhynchus brandti (Coleoptera: Curculionidae).

Dormancy is important for overwintering insects to resist and adapt to adverse conditions. Dormancy generally contains quiescence and diapause. Eucryptorrhynchus brandti Harold (Coleoptera: Curculionidae), tree-of-heaven trunk weevil (TTW), is a destructive pest and highly host-specific to Ailanthus altissima in China. TTW has one generation per year and overwinters as both larvae and adults. In this study, to examined dormancy type of adults and find a method to store overwintering adults, we collected adults from 20 October 2018 to 13 March 2019. We studied the behavior and reproductive development of adults under field cold conditions for 0 and 10 d and laboratory warm conditions for 5 and 10 d. We recorded developing eggs in females, and the clarity of the testis edge, the yellow point in the testis lobe, the ratio of the inner content in the accessory gland, and the accessory gland color in males. Adults transferred from the field to the laboratory had resumed reproductive development directly. Results indicated that the dormancy type of TTW adults was quiescence. Adults stored in the field were still in a dormant state and the field-storage method was effective. Current study provided basic data for controlling overwintering TTW adults and solve the storage of insect sources during the winter.

Functional relationship between woody plants and insect communities in response to Bursaphelenchus xylophilus infestation in the Three Gorges Reservoir region.

To study the effect of the invasion of Bursaphelenchus xylophilus on the functional relationship between woody plants and insect communities, the populations of tree species and insect communities were investigative in the Masson pine forests with different infestation durations of B. xylophilus. In this study, the number of Pinus massoniana began to decrease sharply, whereas the total number of other tree species in the arboreal layer increased gradually with the infestation duration of B. xylophilus. The principal component analysis ordination biplot shows that there was a significant change in the spatial distribution of woody plant species in different Masson pine forest stands. Additionally, a total of 7,188 insect specimens were obtained. The insect population showed an upward trend in stand types with the increase of pine wilt disease infection periods, which demonstrated that the insect community had been significantly affected by the invasion of B. xylophilus. However, the insect diversity indexes were not significantly different among Masson pine forest stands. The structure of insect functional groups changed from herbivorous (He) > omnivorous (Om) > predatory (Pr) > parasitic (Pa) > detritivorous (De) in the control stand to He > Pa > Om, De > Pr after B. xylophilus infestation in the forests. The results showed that the populations of He, Pa, and De increased after the invasion of B. xylophilus, but the populations of Pr decreased. Moreover, the redundancy analysis ordination biplots reflected the complicated functional relationship between woody plant communities and insects after the invasion of B. xylophilus. The present study provides insights into the changes in the community structure of woody plants and insects, as well as the functional relationship between woody plant communities and insect communities after invasion of B. xylophilus.

Comparison and Functional Analysis of Chemosensory Protein Genes From Eucryptorrhynchus scrobiculatus Motschulsky and Eucryptorrhynchus brandti Harold.

The tree-of-heaven root weevil (Eucryptorrhynchus scrobiculatus) and the tree-of-heaven trunk weevil (Eucryptorrhynchus brandti) are closely related species that monophagously feed on the same host plant, the Ailanthus altissima (Mill) Swingle, at different locations. However, the mechanisms of how they select different parts of the host tree are unclear. As chemosensory systems play important roles in host location and oviposition, we screened candidate chemosensory protein genes from the transcriptomes of the two weevils at different developmental stages. In this study, we identified 12 candidate chemosensory proteins (CSPs) of E. scrobiculatus and E. brandti, three EscrCSPs, and one EbraCSPs, respectively, were newly identified. The qRT-PCR results showed that EscrCSP7/8a/9 and EbraCSP7/8/9 were significantly expressed in adult antennae, while EscrCSP8a and EbraCSP8 shared low sequence identity, suggesting that they may respond to different odorant molecule binding. Additionally, EbraCSP6 and EscrCSP6 were mainly expressed in antennae and proboscises and likely participate in the process of chemoreception. The binding simulation of nine volatile compounds of the host plant to EscrCSP8a and EbraCSP8 indicated that (1R)-(+)-alpha-pinene, (-)-beta-caryophyllene, and beta-elemen have higher binding affinities with EscrCSP8a and lower affinities with EbraCSP8. In addition, there were seven, two, and one EbraCSPs mainly expressed in pupae, larvae, and eggs, respectively, indicating possible developmental-related roles in E. brandti. We screened out several olfactory-related possible CSP genes in E. brandti and E. scrobiculatus and simulated the binding model of CSPs with different compounds, providing a basis for explaining the niche differentiation of the two weevils.

Structural comparison of the rostra of two species of weevils coexisting on Ailanthus altissima: the response to ecological demands of egg deposition.

BACKGROUND: Elongated rostra play an important role in the egg-laying of weevils, and its emergence plays a key role in the adaptive radiation of weevils. Eucryptorrhynchus scrobiculatus Motschulsky and E. brandti Harold co-occur on the same only host Ailanthus altissima, while their oviposition sites are different. In order to understand the adaptation between the rostra of the two weevils and their oviposition sites, the structural differentiation of the rostra in E. scrobiculatus and E. brandti was compared. RESULTS: The present study reveals that: (1) The rostra length of E. scrobiculatus and E. brandti was found to be correlated with body size, larger weevils have a correspondingly longer rostrum. The increase of rostra length may be a byproduct of larger weevils. (2) There were significant differences in the external shape of the two rostra, especially the shape of the mandibles of the mouthparts at the apex of the rostra used to excavate an oviposition cavity. (3) There was no difference in the size of the abductor muscles that control the extension of the mandibles, but there were significant differences in the size of the adductor muscles that control the contraction of the mandibles. CONCLUSIONS: These structural differences reflect the functional potential ovipositional tactics of rostra, which is considered to be a response to the ecological demands of egg deposition, and also provide new insights into the coexistence of two weevil species in the same host A. altissima.

Opportunities to improve China's biodiversity protection laws.

Since 1989, China has established a system of powerful laws and regulations aimed to preserve its rich natural flora and fauna. However, this legislative framework still has shortcomings, in terms of sentencing standards across related crimes and the extent of scientific basis for sentences. Here, we review Chinese biodiversity protection laws and some example cases with the goal of suggesting ways to increase law compliance and thus better protect biodiversity. In particular, our suggestions involve regular updates of threat assessments based on scientific evidence including herbaceous plants, fungi and algae; considering ecological differences among the species groups and ensuing ecological damage and financial profit gained; and a differentiation of punishment between organized and individual crimes, with a preference for custodial sentences for the former and monetary fines for the latter, to comply better with international standards and to minimize the incentive to engage in such conduct.

Developing traps for the overwintering tree-of-heaven weevils Eucryptorrhynchus scrobiculatus and E. brandti (Coleoptera: Curculionidae).

BACKGROUND: The tree-of-heaven root and trunk weevils, Eucryptorrhynchus scrobiculatus (TRW) and E. brandti (TTW) are important boring pests of Ailanthus altissima in China. Overwintering TRW and TTW adults can continue to damage A. altissima trees and reproduce after they emerge from the soil each year. There is a lack of studies regarding the development of control measures during the overwintering period. A polyethylene terephthalate bottle trap (PETBT) and corrugated paper box trap (CPBT) were evaluated for their ability to trap TRW and TTW adults descending from trees to overwinter in the soil in Wutongshu and Shabatou locations in Ningxia, China. RESULTS: Significantly more overwintering TRW adults were caught by the PETBT than the CPBT. There were no significant differences in the TTW captures in PETBT when compared to the CPBT. Both PETBT and CPBT have little effect on trapping overwintering TTW adults. Further trapping studies showed that PETBT treatments significantly reduced overwintering TRW numbers caught in population monitoring trap nets placed on the A. altissima tree trunks (an average annual reduction of 42% than the untreated trees). CONCLUSION: These results demonstrate that PETBT is effective for trapping overwintering TRW adults. PETBT could be recommended as an alternative for managing TRW in the framework of integrated pest management. © 2021 Society of Chemical Industry.

Contrasting Behavioral and Electrophysiological Responses of Eucryptorrhynchus scrobiculatus and E. brandti (Coleoptera: Curculionidae) to Volatiles Emitted from the Tree of Heaven, Ailanthus altissima.

Eucryptorrhynchus scrobiculatus and E. brandti (Coleoptera: Curculionidae) are host-specific pests of Ailanthus altissima (Mill.) Swingle (Sapindales: Simaroubaceae), causing extensive damage to the host. There are no effective attractants available for pest management. The main aim of this study was to explore the role of host plant-derived volatiles in the behavioral response of both weevil species. In a field experiment, both weevil species showed positive response to phloem, and there was no preference for phloem associated with healthy or injured trees. Significantly more E. brandti adults responded to the olfactory treatments compared to E. scrobiculatus. In a large-arena experiment, both males and females of E. scrobiculatus significantly preferred phloem from the tree trunk while adults of E. brandti responded in significantly greater numbers to tree limbs than to any other parts of host. Females and males of E. scrobiculatus responded positively to all parts of host tested in the Y-tube bioassay, while E. brandti adults were only attracted by the phloem from healthy and injured trees. There were dissimilar electroantennographic responses to compounds such as 1-hexanol and (1S)-(-)-ß-pinene between the two weevil species. This study represents the first report documenting behavioral and electrophysiological responses of E. scrobiculatus and E. brandti to volatiles from various parts of A. altissima and findings may aid efforts to develop attractants.

Electrophysiological and behavioral responses of red imported fire ants (Hymenoptera: Formicidae) to an essential balm and its components.

BACKGROUND: An over-the-counter medicine product of China known as essential balm effectively repelled red imported fire ants, Solenopsis invicta Buren. However, it was not clear which chemical component(s) accounted for the repellency, and whether they would effectively repel S. invicta in the field. RESULTS: Five components, eucalyptol, camphor, menthol, methyl salicylate, and eugenol, were identified in essential balm using gas chromatography-mass spectrometry (GC-MS). Each component elicited concentration-dependent electroantennography (EAG) response. Under field conditions, all components showed repellency against foraging ants. Interestingly, foraging ants managed to access the food items placed on a surface smeared with eucalyptol, camphor, menthol, or methyl salicylate by depositing soil particles on the surface and then walking on soil particles. However, they failed to do so when the surface was smeared with eugenol. Repellency of eugenol lasted for > 24 h, which was much longer than that of the other four components of essential balm and is comparable to that of N,N-diethyl-m-toluamide (DEET), the standard for insect repellants. CONCLUSION: Olfactory response of S. invicta to all five components of the essential balm was confirmed. Each component showed repellency against S. invicta workers in the field. However, only eugenol significantly suppressed both foraging and particle-covering behavior within 24 h. The repellent effect of eugenol lasted much longer than the other four components. Particle-covering behavior has been largely ignored in studying fire ant repellants. Our study demonstrated that it is necessary to consider such behaviors in ant repellent bioassays in the future. © 2020 Society of Chemical Industry.