Field effects of oxygenated monoterpenes and estragole combined with pheromone on attraction of Ips typographus and its natural enemies

Introduction: Central European Norway spruce monocultures face Ips typographus outbreaks due to decreasing resistance. These beetles use volatile compounds to communicate and select suitable host trees. Spruce trees, beetles, and their symbiotic ophiostomatoid fungi emit oxygenated monoterpenes, including 1,8-cineole, alpha-terpineol, camphor, carvone, terpinen-4-ol, isopinocamphone, and pinocamphone, and the phenylpropanoid estragole, particularly in the infestation phase. These compounds trigger strong responses in I. typographus antennae, motivating our field study.Objective: This study aimed to assess how adding these compounds to the aggregation pheromone of Ips typographus modulates the attraction of this bark beetle and its natural enemies.Methods: In combination with I. typographus pheromone, estragole, 1,8-cineole, (+/-)-camphor, (-)-carvone, alpha-terpineol, (-)-terpinen-4-ol, (+)-pinocamphone, and (+)-isopinocamphone at low, medium, and high doses were tested in pheromone traps at two sites in the Czech Republic.Results: All 1,8-cineole doses and the high estragole dose acted as anti-attractants for I. typographus, whereas all (+)-isopinocamphone doses enhanced their attraction to pheromone. Catches of natural enemies, the Staphylinidae and Pteromalidae, varied by location.Conclusion: 1,8-cineole, isopinocamphone, and estragole may play vital roles in tritrophic interactions among spruce trees, and I. typographus and its natural enemies, and these compounds may be developed into new or enhanced semiochemical-based pest control methods

Host-dependent larval migration and parasitism risk in a polyphagous moth

Female herbivorous insects are expected to oviposit on the host plant providing the best performance of the offspring. However, in some insects the larvae are mobile and are not totally dependent on the mother's choice. They can change host plant when conditions for development or exposure to natural enemies vary between individual plants within a patch. Here we study larval migration and preference between two host plants, cotton and alfalfa, in the Egyptian leafworm, Spodoptera littoralis (Boisduval) (Lepidoptera: Noctuidae). Previous studies have shown that although larval performance is better on cotton, females prefer to oviposit on alfalfa, and this preference corresponds to a higher survival of the eggs. In this study, S. littoralis larvae showed directed movement between host plants and were found to prefer alfalfa over cotton in field test in Egypt, as well as in laboratory selection of feeding site assays. To determine effects by natural enemies, the parasitism rates and various life-history traits were measured for one larval parasitoid, Microplitis rufiventris Kokujev (Hymenoptera: Braconidae), on several host plants including cotton and clover (equivalent host plant to alfalfa). Overall, parasitism was higher on cotton and parasitoid performance (cocoon mass, adult longevity, and female egg load) was better on cotton compared to clover. This fact suggests an enemy-free space on clover and alfalfa, as parasitism rate is higher on cotton, and the parasitoid performance is also better on cotton-fed larvae.Larval migration was investigated in the Egyptian leafworm, Spodoptera littoralis (Lepidoptera: Noctuidae). Larvae showed directed movement between host plants and were found to prefer alfalfa over cotton in field and laboratory tests, even though larval performance is better on cotton. However, parasitism rates were higher and parasitoid performance (cocoon mass, adult longevity, and female egg load) was better for the larval parasitoid Microplitis rufiventris (Hymenoptera: Braconidae) on cotton, suggesting enemy-free space on alfalfa.imag

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

Direct seeding for restoration of mixed oak forests: Influence of distance to forest edge, predator-derived repellent and acorn size on seed removal by granivorous rodents

Restoration of mixed oak forest in northern temperate regions is important for biodiversity and for adaptation of forest management to climate change. Direct seeding has been considered as a cost-effective method for the assisted regeneration of oaks. However, removal of acorns by granivorous rodents hinders its application. Patterns of acorn removal can vary with habitat type, presence of predators, and acorn size. In this field study, we tested the effects of distance to forest edge, predator excrement as a repellent, and acorn size on the performance of direct-seeded oak in southern Sweden. The experiment was carried out at two different sites. Our hypotheses were: (i) in clear-cuts, acorn removal is higher closer to the forest edge compared to open areas, (ii) treatment with predator excrement reduces acorn removal without affecting germination, and (iii) larger acorns will be removed more readily than smaller acorns. Finally, we also investigated if seedlings from larger acorns survive better and grow taller compared to smaller acorns in relation to habitat structure and predator excrement as repellent. Acorn removal differed at the two sites. At Skrylle (older clear-cut), no forest edge effects were found. At Klåveröd (new clear-cut site), our results suggest that removal of acorns was higher in the open clear-cut due to the presence of large slash piles. Less vegetation coverage correlated with lower rates of acorn removal, except near slash piles. Our results therefore suggest that forest managers should reduce understory vegetation and avoid slash piles during direct seeding of oak. Mink excrement failed to protect acorns against removal, but did not reduce acorn germination. Bigger acorns produced taller seedlings but did not enhance overall woodland establishment due to their higher removal rate. Our results further suggest a potential food satiation effect due to an extra supply of additional food. Satiation manipulation might potentially be a method of mitigating the impact of seed predation by granivorous rodents during the direct seeding of oak, but such an approach requires further research before it could be recommended

Semiochemicals produced by fungal bark beetle symbiont Endoconidiophora rufipennis and the discovery of an anti-attractant for Ips typographus

Bark beetles vector symbiotic fungal species into their host trees during mass attacks. The symbiotic relationship with blue stain fungi of the Ascomycetes, including genera of Endoconidiophora (syn. = Ceratocystis), promotes successful establishment whereby the microbes help to overcome the host trees' defence and degrade toxic resins. This is the first study to evaluate both the volatile emissions from an insect-associated blue stain fungus over time and the insect response in a field trapping experiment. Volatile emissions from isolates of Endoconidiophora rufipennis (ER) were collected by solid-phase microextraction (SPME) and analysed by gas chromatography-mass spectroscopy (GC-MS) over a period of 30 days. This virulent North American fungus is closely related to E. polonica, a symbiotic fungus known from Eurasian spruce bark beetle Ips typographus.Nine volatiles were emitted by ER in substantial amounts: isoamyl acetate, sulcatone, 2-phenethyl acetate, geranyl acetone, geranyl acetate, citronellyl acetate, (R)- and (S)-sulcatol, and (R)-sulcatol acetate. A late peaking compound was geranyl acetone. In the field trapping experiment, three of the fungal volatiles (geranyl acetone, 2-phenethyl acetate and sulcatone) were tested in combination with a synthetic aggregation pheromone for I. typographus. Traps with geranyl acetone attracted lower numbers of I. typographus compared to traps with 2-phenethyl acetate, sulcatone or the pheromone alone as a control. The results showed that geranyl acetone acts as an anti-attractant and may act naturally on I. typographus as a cue from an associated fungus to signal an overexploited host

Interactions among Norway spruce, the bark beetle Ips typographus and its fungal symbionts in times of drought

Resilience and functionality of European Norway spruce forests are increasingly threatened by mass outbreaks of the bark beetle Ips typographus promoted by heat, wind throw and drought. Here, we review current knowledge on Norway spruce and I. typographus interactions from the perspective of drought-stressed trees, host selection, colonisation behaviour of beetles, with multi-level effects of symbiotic ophiostomatoid fungi. By including chemo-ecological, molecular and behavioural perspectives, we provide a comprehensive picture on this complex, multitrophic system in the light of climate change. Trees invest carbon into specialised metabolism to produce defence compounds against biotic invaders; processes that are strongly affected by physiological stress such as drought. Spruce bark contains numerous terpenoid and phenolic substances, which are important for bark beetle aggregation and attack success. Abiotic stressors such as increased temperatures and drought affect composition, amounts and emission rates of volatile compounds. Thus, drought events may influence olfactory responses of I. typographus, and further the pheromone communication enabling mass attack. In addition, I. typographus is associated with numerous ophiostomatoid fungal symbionts with multiple effects on beetle life history. Symbiotic fungi degrade spruce toxins, help to exhaust tree defences, produce beetle semiochemicals, and possibly provide nutrition. As the various fungal associates have different temperature optima, they can influence the performance of I. typographus differently under changing environmental conditions. Finally, we discuss why effects of drought on tree-killing by bark beetles are still poorly understood and provide an outlook on future research on this eruptive species using both, field and laboratory experiments

Non-host volatile blend optimization for forest protection against the European Spruce Bark Beetle, Ips typographus

Conifer feeding bark beetles (Coleoptera, Curculionidae, Scolytinae) pose a serious economic threat to forest production. Volatiles released by non-host angiosperm plants (so called non-host volatiles, NHV) have been shown to reduce the risk of attack by many bark beetle species, including the European spruce bark beetle, Ips typographus. However, the most active blend for I. typographus, containing three green leaf volatiles (GLVs) in addition to the key compounds trans-conophthorin (tC) and verbenone, has been considered too expensive for use in large-scale management. To lower the cost and improve the applicability of NHV, we aim to simplify the blend without compromising its anti-attractant potency. Since the key compound tC is expensive in pure form, we also tested a crude version: technical grade trans-conophthorin (T-tC). In another attempt to find a more cost effective substitute for tC, we evaluated a more readily synthesized analog: dehydroconophthorin (DHC). Our results showed that 1-hexanol alone could replace the three-component GLV blend containing 1- hexanol, (3Z)-hexen-1-ol, and (2E)-hexen-1-ol. Furthermore, the release rate of tC could be reduced from 5 mg/day to 0.5 mg/day in a blend with 1-hexanol and (–)-verbenone without compromising the anti-attractant activity. We further show that T-tC was comparable with tC, whereas DHC was a less effective anti-attractant. DHC also elicited weaker physiological responses in the tC-responding olfactory receptor neuron class, providing a likely mechanistic explanation for its weaker anti-attractive effect. Our results suggest a blend consisting of (–)-verbenone, 1-hexanol and technical transconophthorin as a cost-efficient anti-attractant for forest protection against I. typographus

Unravelling the gut bacteriome of Ips (Coleoptera: Curculionidae: Scolytinae): identifying core bacterial assemblage and their ecological relevance

Bark beetles often serve as forest damaging agents, causing landscape-level mortality. Understanding the biology and ecology of beetles are important for both, gathering knowledge about important forest insects and forest protection. Knowledge about the bark beetle gut-associated bacteria is one of the crucial yet surprisingly neglected areas of research with European tree-killing bark beetles. Hence, in this study, we survey the gut bacteriome from five Ips and one non-Ips bark beetles from Scolytinae. Results reveal 69 core bacterial genera among five Ips beetles that may perform conserved functions within the bark beetle holobiont. The most abundant bacterial genera from different bark beetle gut include Erwinia, Sodalis, Serratia, Tyzzerella, Raoultella, Rahnella, Wolbachia, Spiroplasma, Vibrio, and Pseudoxanthomonas. Notable differences in gut-associated bacterial community richness and diversity among the beetle species are observed. Furthermore, the impact of sampling location on the overall bark beetle gut bacterial community assemblage is also documented, which warrants further investigations. Nevertheless, our data expanded the current knowledge about core gut bacterial communities in Ips bark beetles and their putative function such as cellulose degradation, nitrogen fixation, detoxification of defensive plant compounds, and inhibition of pathogens, which could serve as a basis for further metatranscriptomics and metaproteomics investigations

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

Testing the Efficiency of the Push-and-Pull Strategy during Severe Ips typographus Outbreak and Extreme Drought in Norway Spruce Stands

Protection of Norway spruce stands using anti-attractants was tested during an outbreak of bark beetles (Ips typographus) in their spring flight. The aims of this study were as follows: (1) to test the proposed experimental design for tree protection; (2) to evaluate height-specific alternatives for dispenser installation on trees; and (3) to evaluate the efficiency of tree protection measures using anti-attractants under bark beetle infestation and drought stress. The experiment was conducted at the forest edges adjacent to recent clearcuts on 10 blocks in the eastern Czech Republic. Each block had three adjacent experimental areas, with 20 trees growing in two rows at the recently cut forest edge (10 trees per row). In front of a block in each of the three areas, four pheromone traps were installed. The treatment area was protected by anti-attractants. The second area served as a so-called switch area, where beetles from the treatment area, as the outflux redirected from the anti-attractant, would start new attacks if not caught in nearby pheromone traps. The third area was a control. We attached anti-attractant tube dispensers on each tree trunk of the treated area at two heights. The results suggest a redirecting effect of anti-attractants, pushing beetles into the switch area and causing subsequent attacks, which was greater than in areas containing treated trees. There was no difference between two dispensers placed at 1 and 8 m height and both at 1 m. A switching effect of beetle attacks occurring outside of the treated areas was observed. Mounting anti-attractant dispensers on tree trunks at one low position above the ground can be substantially less labour-intensive and as efficient as positioning them at two different heights. For areas affected by severe drought and extremely dense bark beetle populations, the use of anti-attractants did not prove effective