Fear effects on bank voles (Rodentia: Arvicolinae): testing for repellent candidates from predator volatiles

BACKGROUND Arvicolinae rodents are known pests causing damage to both agricultural and forest crops. Today, rodenticides for rodent control are widely discouraged owing to their negative effects on the environment. Rodents are the main prey for several predators, and their complex olfactory system allows them to identify risks of predation. Therefore, the potential use of predators' scents as repellents has gained interest as an ecologically based rodent control method. In a two-choice experiment, we investigated the potential repellent effects of five synthetic predator compounds: 2-phenylethylamine (2-PEA), 2-propylthietane (2-PT), indole, heptanal and 2,5-dihydro-2,4,5-trimethylthiazoline (TMT), at 1% and 5% doses, using the bank vole (Myodes glareolus) as a rodent model. RESULTS The compound 2-PEA reduced both the food contacts and the time spent by voles in the treatment arm compared to the control arm. Likewise, 2-PT-treated arms reduced the food contacts, and the voles spent less time there, although this latter difference was not significant. Indole also showed a tendency to reduce the time spent at the treatment arm; however, this result was not significant. Unexpectedly, TMT had the reverse effect in showing attractive properties, possibly due to odor cues from differently sized predators and intraguild predation in nature. We found no dose-related effects for any compounds tested. CONCLUSION Our results suggest that the 2-PEA and 2-PT are both effective odor stimuli for triggering reduced food contacts and area avoidance, and they may be good repellent candidates. We suggest further testing of 2-PEA and 2-PT in field experiments to further determine their dose-efficiency as repellents against rodents in more natural environments

Malaria mosquitoes acquire and allocate cattle urine to enhance life history traits

Background: Nutrient acquisition and allocation integrate foraging and life-history traits in insects. To compensate for the lack of a particular nutrient at different life stages, insects may acquire these through supplementary feeding, for example, on vertebrate secretions, in a process known as puddling. The mosquito Anopheles arabiensis emerges undernourished, and as such, requires nutrients for both metabolism and reproduction. The purpose of this study was to assess whether An. arabiensis engage in puddling on cattle urine to obtain nutrients to improve life history traits.Methods: To determine whether An. arabiensis are attracted to the odour of fresh, 24 h, 72 h and 168 h aged cattle urine, host-seeking and blood-fed (48 h post-blood meal) females were assayed in a Y-tube olfactometer, and gravid females assessed in an oviposition assay. Combined chemical and electrophysiological analyses were subsequently used to identify the bioactive compounds in all four age classes of cattle urine. Synthetic blends of bioactive compounds were evaluated in both Y-tube and field assays. To investigate the cattle urine, and its main nitrogenous compound, urea, as a potential supplementary diet for malaria vectors, feeding parameters and life history traits were measured. The proportion of female mosquitoes and the amount of cattle urine and urea imbibed, were assessed. Following feeding, females were evaluated for survival, tethered flight and reproduction.Results: Host-seeking and blood-fed An. arabiensis were attracted to the natural and synthetic odour of fresh and aged cattle urine in both laboratory and field studies. Gravid females were indifferent in their response to cattle urine presence at oviposition sites. Host-seeking and blood-fed females actively imbibed cattle urine and urea, and allocated these resources according to life history trade-offs to flight, survival or reproduction, as a function of physiological state.Conclusions: Anopheles arabiensis acquire and allocate cattle urine to improve life history traits. Supplementary feeding on cattle urine affects vectorial capacity directly by increasing daily survival and vector density, as well as indirectly by altering flight activity, and thus should be considered in future models

Identification of cattle-derived volatiles that modulate the behavioral response of the biting midge culicoides nubeculosus

Identification of host-derived volatiles is an important step towards the development of novel surveillance and control tools for Culicoides biting midges. In this study, we identified compounds from headspace collections of cattle hair and urine that modulate the behavioral response of Culicoides nubeculosus, a research model species with a similar host-range as the vectors of Bluetongue disease and Schmallenberg disease in Europe. Combined gas chromatography and electroantennographic detection (GC-EAD) analysis revealed 23 bioactive compounds, of which 17, together with octanal, were evaluated in a two-choice behavioral assay in the presence of CO2. Decanal, 2-phenylethanal, 1-octen-3-ol, 2-ethylhexanol, 3-methylindole, phenol, and 3-ethylphenol elicited attraction of host seeking C. nubeculosus, whereas heptanal, octanal, nonanal, 3-propylphenol, and 4-propylphenol inhibited the insects’ attraction to CO2, when compared to CO2 alone. 6-Methyl-5-hepten-2-one, 3-methylphenol, 4-methylphenol, and 4-ethylphenol elicited both attraction and inhibition. The behavioral responses were dependent on the concentration tested. Our results show that cattle-derived odors have the potential to be used for the manipulation of the behavior of Culicoides biting midges

Odors Attracting the Long-Legged Predator Medetera signaticornis Loew to Ips typographus L. Infested Norway Spruce Trees

Predatory long-legged flies of the genus Medetera are important, but currently understudied, natural enemies of Scolytinae bark beetles such as Ips typographus. Medetera flies lay eggs on beetle-infested trees, where the developing larvae find their prey, but the chemical cues used by Medetera to locate infested trees are currently unknown. To identify odors attracting Medetera signaticornis, a species in Europe, headspace samples were collected at several time-points through different stages of I. typographus attacks on logs of Norway spruce (Picea abies). The headspace samples were analyzed using combined gas chromatography and mass spectrometry (GC-MS), and gas chromatography coupled with electroantennographic detection (GC-EAD) to determine compounds that stimulate M. signaticornis antennae. Antennae of M. signaticornis males and females were found to detect (-)-cis-verbenol, (+)-trans-verbenol and myrtenol, which are known to be produced by bark beetles. Antennal responses were also observed for verbenene, isoterpinolene, alpha-pinene oxide, camphor, pinocamphone, terpinene-4-ol, myrtenal, borneol, alpha-terpineol, geranyl acetone, and verbenone, which are primarily produced by microorganisms, and alpha-pinene, alpha-fenchene, beta-pinene, camphene, 3-carene, limonene, gamma-terpinene, and terpinolene, known spruce tree compounds. In field experiments testing two synthetic blends containing 18 antennal active and two additional compounds 2-methyl-3-buten-2-ol and ipsdienol we observed significant attraction of M. signaticornis within 24 h. These attractive blends can form the basis for development of Medetera monitoring lures for use in future forest and pest management

Mate recognition and reproductive isolation in the sibling species Spodoptera littoralis and Spodoptera litura

Mate recognition is crucial for reproductive isolation and for maintaining species integrity. Chemosensory-mediated sexual communication with pheromones is an essential component of mate recognition in moths. Confronted with sex pheromone stimuli released from conspecific and closely related heterospecific females, which partially overlap in chemical composition, male moths are under strong selection to recognize compatible mates. Here, we investigated the role of pheromone signals in premating communication in the sibling species Spodoptera littoralis and S. litura (Lepidoptera, Noctuidae). Further, we measured the reproductive consequence of conspecific vs. heterospecific matings. Both species use Z9,E11–14:Ac as the major pheromone compound, and the 11-component blend found in pheromone glands of S. littoralis comprises the compounds found in S. litura. Accordingly, S. littoralis and S. litura males readily responded to conspecific and heterospecific calling females in no-choice behavioral tests. In contrast, in a dual-choice test, S. littoralis males choose conspecific calling females, whereas S. litura males did not discriminate between conspecific and heterospecific females. In S. littoralis females, heterospecific matings had a negative fitness effect as compared to conspecific matings. Female longevity, egg-laying and hatching of larvae were significantly reduced by matings with heterospecific males. Reciprocal crossings, between S. litura females and S. littoralis males, were prevented by genital morphology, which is consistent with reduced heterospecific attraction of S. littoralis males in a dual-choice assay. On the other hand, matings between S. littoralis females and S. litura males, under a no-choice situation, show that interspecific matings occur in zones of geographical overlap and corroborate the idea that mate quality, in these closely related species, is a continuous and not a categorical trait

Flight attraction of Spodoptera littoralis (Lepidoptera, Noctuidae) to cotton headspace and synthetic volatile blends

The insect olfactory system discriminates odor signals of different biological relevance, which drive innate behavior. Identification of stimuli that trigger upwind flight attraction toward host plants is a current challenge, and is essential in developing new, sustainable plant protection methods, and for furthering our understanding of plant-insect interactions. Using behavioral, analytical and electrophysiological studies, we here show that both females and males of the Egyptian cotton leafworm,Spodoptera littoralis(Lepidoptera, Noctuidae), use blends of volatile compounds to locate their host plant, cotton,Gossypium hirsutum(Malvales, Malvaceae). FemaleS. littoraliswere engaged in upwind orientation flight in a wind tunnel when headspace collected from cotton plants was delivered through a piezoelectric sprayer. Although males took off toward cotton headspace significantly fewer males than females flew upwind toward the sprayed headspace. Subsequent assays with antennally active synthetic compounds revealed that a blend of nonanal, (Z)-3 hexenyl acetate, (E)-β-ocimene, and (R)-(+)-limonene was as attractive as cotton headspace to females and more attractive to males. Two compounds, 4,8-dimethyl-1,3(E),7-nonatriene (DMNT) and (R)-(−)-linalool, both known plant defense compounds may have reduced the flight attraction of both females and males; more moths were attracted to blends without these two compounds, however, other compounds such as benzaldehyde may also be behavioral antagonists. Our findings provide a platform for further investigations on host plant signals mediating innate behavior, and for the development of novel insect plant protection strategies againstS. littoralis

Predator odor can reduce acorn removal by granivorous rodents in mixed oak forest stands

Developing better practices for rodent pest control is of high importance to reduce damage during forest restoration and in crop production. For example, during direct seeding with large and highly attractive seeds such as acorns, most seeds will disappear due to consumption or dispersal if not protected. An unexplored concept in reducing rodent damage is the use of repellents derived from predators. We tested the efficiency of three volatile compounds (2-propylthietane, 2-phenylethylamine and indole) associated with predators as rodent repellent candidates and scored the reduction of acorn (Quercus petraea) removal at two field sites in southern Sweden. We further investigated at what distance (5 cm, 10 cm, and 15 cm) from the odor source the odors were efficient in lowering the removal of acorns. Removal was lowest with 2-propylthietane (25-45 % of acorn removal), followed by 2-phenylethylamine (75-95% acorn removal) at 5 cm. Indole failed to decrease acorn removal and did not differ significantly from the control treatments. In the control treatments, almost all acorns (95-100%) were removed from the plots during the 48-hour sessions at both sites, and the acorns were removed faster than in the other treatments. Removal increased with distance from the 2-propylthietane odor dispenser at both field sites. Here, the lowest acorn removal occurred at 5 cm from the odor disperser. Our short time experiment shows for the first time the potential of using 2-propylthietane for short-term protection of acorns from foraging by granivorous rodents in oak woodlands. To help increase the reliability of direct seeding as a method for regenerating and restoring forests, future studies should investigate whether the range and longevity in the field of predator odor formulations can be improved

Antennal and maxillary palp morphology, and sensillar equipment, of the spruce bark beetle predators, Medetera signaticornis and Medetera infumata (Diptera: Dolichopodidae)

Many long-legged Medetera flies are natural enemies of bark beetle pests, which they detect using ol-factory cues, likely through olfactory sensilla on the antennae and maxillary palps. Morphological characterisation of olfactory sensilla among insects can provide a basis for future taxonomic, phyloge-netic or electrophysiological studies. Scanning electron microscopy was used to describe the morphology of olfactory organs and sensillar equipment of Medetera signaticornis and M. infumata. Three different olfactory sensillum types were found in both fly species, sensilla trichodea, s. basiconica and grooved pegs. Based on size and wall structure, s. trichodea and s. basiconica were categorised into different subtypes. Sharp-tipped curved s. trichodea, and small, large and thin s. basiconica were found on the antennal postpedicel of M. signaticornis adults, while grooved s. basiconica were found in M. infumata. The density of sharp-tipped long s. trichodea was significantly higher in males compared to females, and in M. signaticornis compared to M. infumata. Long-grooved s. basiconica were found grouped in a small pit on the maxillary palps of both species. Comparison of our results with the limited available ecological data suggests that differences in numbers of specific sensillum types may reflect adaptations related to olfactory-driven behaviours such as host seeking.(c) 2022 The Author(s). Published by Elsevier Ltd. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/)

Ecological and Phylogenetic Relationships Shape the Peripheral Olfactory Systems of Highly Specialized Gall Midges (Cecidomiiydae)

Insects use sensitive olfactory systems to detect relevant host volatiles and avoid un-suitable hosts in a complex environmental odor landscape. Insects with short lifespans, such as gall midges (Diptera: Cecidomyiidae), are under strong selection pressure to detect and locate suitable hosts for their offspring in a short period of time. Ephemeral gall midges constitute excellent models for investigating the role of olfaction in host choice, host shift and speciation. Midges mate near their site of emergence and females migrate in order to locate hosts for oviposition, thus females are expected to be more responsive to olfactory cues emitted by the host compared to males. In this study, we explored the correlation between host choice and the func-tion of the peripheral olfactory system in twelve species of gall midges, including species with close phylogenetic relationships that use widely different host plants and more distantly related gall midge species that use similar hosts. We tested the anten-nal responses of males and females of the twelve species to a blend of 45 known insect attractants using coupled gas chromatographic–electroantennographic detec-tion. When the species-specific response profiles of the gall midges were compared to a newly generated molecular-based phylogeny, we found they responded to the com-pounds in a sex- and species-specific manner. We found the physiological response profiles of species that use annual host plants, and thus have to locate their host every season, are similar for species with similar hosts despite large phylogenetic distances. In addition, we found closely related spe-cies with perennial hosts demonstrated odor response profiles that were consistent with their phylogenetic history. The ecology of the gall midges affects the tuning of the peripheral olfactory system, which in turn demonstrates a correlation between olfaction and speciation in the context of host use