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Key message The dog detection allows timely removal by sanitation logging of first beetle-attacked trees before offspring emergence, preventing local beetle increases. Detection dogs rapidly learned responding to synthetic bark beetle pheromone components, with known chemical titres, allowing search training during winter in laboratory and field. Dogs trained on synthetics detected naturally attacked trees in summer at a distance of > 100 m. Context An early detection of first beetle-attacked trees would allow timely sanitation felling before offspring emergence, curbing local beetle increase. Aims We tested if detection dogs, trained off-season on synthetic pheromone components from Ips typographus, could locate naturally bark beetle–infested spruce trees. Methods Indoor training allowed dogs to discriminate between the infestation odours (target) and natural odours (non-target) from the forest. Odour stimuli were shown by chemical analysis to be bioactive at extremely low-levels released (−4 ng/15 min) in the laboratory. Results Detection dogs, trained to recognise four different synthetic pheromone compounds in the wintertime, were able to detect naturally infested spruce trees unknown to humans the following summer. The dog-handler pairs were able to detect an infested spruce tree from the first hours of beetle attack until several weeks after first attack, long before discolouration of the crown. Trained sniffer dogs detected infested spruce trees out to ≥ 100 m, as measured by GPS-collar tracks. Conclusion Dog-handler pairs appear to be more efficient than humans alone in timely detecting bark beetle infestations due to the canine’s ability to cover a greater area and detect by olfaction infestations from a far longer distance than can humans.

In this proof of concept study, we report the off season training of two detection dogs on a series of synthetic semiochemicals associated with Ips typographus pest bark beetle infestations of spruce trees. Scent detection training allowed dogs to discriminate between physiologically-relevant infestation (target) odours, quantified by GC-MS using extracted ion chromatogram to be bio-active at levels of < 10−4 ng /15 min or lower, and natural non-target odours that might be encountered in the forest. Detection dogs trained to recognize four different synthetic pheromone compounds in the winter time, well before beetle flight, were able to detect natural infested spruce trees unknown to humans the following summer. The trained detection dogs were able to detect an infested spruce tree from the first hour of bark beetle attack until several weeks after the attack. Trained detection dogs appear to be more efficient than humans in detecting early bark beetle infestations because the canines ability to cover a greater area and by olfaction detect infestations from a far greater distance than can humans. Infested spruce trees could be detected by trained detection dogs out to more than 100 m.Key MessageDetection dogs were rapidly trained to locate release of synthetic bark beetle pheromone componentsSynthetics allowed dog training off-season both in laboratory and fieldDogs trained on synthetics detected naturally target pest insect attacked trees at a distance of more than 100 m.The method allows rapid removal of single, first attacked trees before offspring emergence, thus curbing local pest increase and lowering spread of attacks in the landscap

Eastern Palearctic conifers are subject to frequent bark beetle outbreaks. However, neither the species responsi- ble nor the semiochemicals guiding these attacks are well understood. Two high-mountain Ips species on Qinghai spruce, Picea crassifolia, I. shangrila and I. nitidus, are typi- cal in this regard. Six synthetic candidate pheromone compo- nents that we earlier identified from hindguts of unmated males of these two Ips species were tested for field activity in Qinghai province, P. R. China. For I. nitidus, racemic ipsdienol ((±)-Id) could replace the naturally-produced blend of enantiomers containing 74 % (-)-(S)-Id (74:26 S:R), in attractive ternary or binary blends. In contrast, sympatric I. shangrila were attracted mainly to blends including Id of opposite chirality, 97 %-(+)-(R)-Id. Of the verbenols, (-)- trans-verbenol was inactive for I. nitidus or inhibitory for I. shangrila ,b ut ( -)-cis-verbenol (cV) was a key component of the pheromone in both species. Two fully factorial experiments demonstrated that (±)-Id, cV, and 2-methyl-3- buten-2-ol (MB) are components of the aggregation phero- mone of I. nitidus, whereas only (+)-Id and cV are essential components of the aggregation pheromone of I. shangrila. While MB is not necessary for attraction of I. shangrila ,i t is an active antagonist and likely functions in species isolation. A review of the pheromone production and responses in Pa- learctic Ips and Pseudoips showed that cV is more common than methylbutenols, and both elicit qualitatively variable re- sponses. Ipsdienol is the most common component with var- iable chirality, and is a necessary, but often not sufficient, factor for determining pheromone specificity.

1. The sense of smell is crucial for fitness of most animals, enabling them to find mates, food and egg laying sites and to stay away from danger. Hence, odour molecules are detected by sensitive and specific olfactory sensory neurons (OSNs). In insects, the OSNs are stereotypically grouped into olfactory sensilla located mainly on the antennae. The functional significance of this co-localization principle is poorly understood, but it has been hypothesized that it allows for coincidence detection of odour filaments, improving discrimination of closely separated odour sources. 2. Using an insect in its natural environment, we conducted the first experimental test of the hypothesis. We manipulated the distance between odour sources of an attractive pheromone and either of two host-derived attraction antagonists (1,8-cineole and verbenone) and investigated the effect on trap catches of the bark beetle, Ips typographus (Coleoptera). 1,8-Cineole is detected by an OSN co-localized with an OSN for one of the pheromone components, while verbenone is detected by OSNs in other sensilla, not co-localized with pheromone OSNs. 3. Consistent with the hypothesis, trap catch increased with distance between odour sources more for 1,8-cineole than for verbenone. The strongest effect was found among the males, that is the sex that first locates and attacks the host tree. 4. Our data from the beetle provide, for the first time, direct experimental support for the hypothesis that co-localization of OSNs in sensilla improves the discrimination of closely separated odour sources. Thus, selection for improved odour source discrimination could well be one of the factors explaining the strict co-localization of OSNs that is seen across the Insecta class. (Less)

In order to locate mates, food, and oviposition sites, insects mainly rely on volatile cues released by their sexual partners, food sources, and host and non-host plants. Calling, mating, and oviposition behaviors, as well as fecundity and longevity, of newly emerged Spodoptera littoralis (Bois.) moths were recorded in the presence of volatiles from leaves of a host plant, Gossypium hirsutum (cotton) and two non-host plants, Adhatoda vasica (Av) or Picea abies (spruce), either alone or in host/non-host combinations. Females exposed to cotton volatiles started calling earlier than females exposed to non-host plant volatiles (NHV), or the blank control. Likewise, moth pairs exposed to cotton volatiles started mating earlier than the other treatments. The period of calling in females alone was longer than females kept with males, having the opportunity to mate. However, the callings, as well as mating durations in the moth pairs, in different treatments were not different. Longevity was decreased either in the absence of cotton or the presence of Av, and spruce leaves. Fecundity was reduced in moths exposed to a combination of spruce and cotton. The effect of NHV on attraction of 2-3-day-old male moths towards a pheromone (Ph) source was studied in a wind tunnel. In the no-choice assay, more males arrived at close approach and landed on the Ph source when the host plant, cotton, was offered in the background as compared to the non-hosts. In the dual-choice assay, more males landed on the Ph source in front of the host plant compared to the Ph source in front of non-hosts. Gas chromatography-electroantennographic detection on female S. littoralis revealed five antennally active compounds in headspace collections of spruce and three compounds in Av.

Bark beetles face challenges and trade-offs during host selection, imposed by lethal tree defences, lower nutrition and higher competition in less well-defended trees, scarcity and ephemeral distribution of susceptible hosts, limitation of suitable hosts to one beetle generation, and relatively short lifespan and vulnerability of adults during host searching. Beetles contend with these challenges by using multiple, integrated sensory modalities, and sequential decision making. They incorporate both negative and positive information at multiple scales to locate susceptible hosts across large and heterogeneous landscapes. Some of the olfactory sensory neurons that convey non-host signals are relatively broadly tuned, whereas those that underlie intraspecific communication and host quality assessment are more specific, an arrangement that maximizes the use of antennal space while retaining high fidelity of detection channels strongly linked to survival and reproduction. The pertinent co-localization of neurons within sensilla provides the means for odour mixture processing in the periphery and enhances odour source discrimination and evaluation of chemical ratios in host tissue. Bark beetles show high behavioural plasticity in their orientation sequences, which allows them to adjust to environmental variability. This plasticity is modulated by environmental, genetic, and gene × environment drivers. Behavioural plasticity allows individuals to best realize the benefits that can be derived from pheromone-mediated cooperative attacks when populations are high. Cross-scale linkages among neurons, sensilla, orientation sequences, populations, and habitat structure underlie the landscape-scale environmental and socioeconomic impacts bark beetles exert. They also underlie rapid outbreaks in response to human-induced environmental alterations, such as climate change, habitat manipulation, and global transport.

Orientation for insects in olfactory landscapes with high semiochemical diversity may be a challenging task. The partitioning of odor plumes into filaments that are interspersed with pockets of 'clean air' may help filament discrimination and upwind flight to attractive sources in the face of inhibitory signals. We studied the effect of distance between odor sources on trap catches of the beetle, Ips typographus, and the moth, Spodoptera littoralis. Insects were tested both to spatially separated pheromone components [cis-verbenol and 2-methyl-3-buten-2-ol for Ips; (Z,E)-9,11-tetradecadienyl acetate and (Z,E)-9,12-tetradecadienyl acetate for Spodoptera], and to separated pheromone and anti-attractant sources [non-host volatile (NHV) blend for Ips; (Z)-9-tetradecenyl acetate for Spodoptera]. Trap catch data were complemented with simulations of plume structure and plume overlap from two separated sources using a photo ionization detector and soap bubble generators. Trap catches of the beetle and the moth were both affected when odor sources in the respective traps were increasingly separated. However, this effect on trap catch occurred at smaller (roughly by an order of magnitude) odor source separation distances for the moth than for the beetle. This may reflect differences between the respective olfactory systems and central processing. For both species, the changes in trap catches in response to separation of pheromone components occurred at similar spacing distances as for separation of pheromone and anti-attractant sources. Overlap between two simulated plumes depended on distance between the two sources. In addition, the number of detected filaments and their concentration decreased with downwind distance. This implies that the response to separated odor sources in the two species might take place under different olfactory conditions. Deploying multiple sources of anti-attractant around a pheromone trap indicated long-distance (meter scale) effects of NHV on the beetle and a potential use for NHV in forest protection.

Stereoisomers of 4-methyl-3-heptanol (MH) are pheromone components of several Scolytus bark beetles. The elm bark beetle Scolytus laevis (Coleoptera: Scolytidae) has in previous studies been caught in traps baited with commercial MH containing all four stereoisomers, but the lure has been considered a weak attractant. In this study, we addressed the question whether stereospecific responses by S. laevis to stereoisomers of MH might contribute to its niche separation from other sympatric Scolytus species. Using GC–MS, we analyzed extracts of hindguts and abdomens from male and female S. laevis and the sympatric S. triarmatus. We also tested all four MH-stereoisomers individually and in combinations in the field to determine their role for S. laevis. All four stereoisomers were synthesized via a boronic ester method with 1,2-dicyclohexylethanediol as chiral director. In addition, the (3S,4R)-stereoisomer of MH was prepared through enantioselective, lipase-mediated transesterification of a mixture of the four stereoisomers of MH. Females of both species contained small amounts of syn-MH, and males contained trace amounts of anti-MH. The anti stereoisomer (3R,4S)-MH was attractive to male and female S. laevis, whereas the syn stereoisomer (3S,4S)-MH acted as an inhibitor or deterrent and reduced the catch when added to the attractive isomer. The syn isomer is the main aggregation pheromone component of the larger and sympatric S. scolytus and possibly also of S. triarmatus. The avoidance response of S. laevis to the (3S,4S)-stereoisomer may reduce interspecific competition for host trees.

BACKGROUND: Field trapping experiments were carried out to evaluate effective trap characteristics for maximising Ips duplicatus(Sahlberg) catches in pheromone-baited traps in China. RESULTS: Window-slot and cross-barrier traps had significantly higher catches than multiple-funnel traps. The colour of window-slot traps showed a significant effect on catches, with dark colours (black and red) being more effective than light colours, especially white and yellow. Window-slot traps at a 1.5‐2.0 m level caught more beetles than those at either ground level (0‐0.5 m) or at 3.5‐4.0 m. Ips duplicatus can be attracted to pheromone-baited traps over a distance of >100 m from the forest edge in an open grassy field. There was a strong diurnal pattern of flight activity, with catches on window-slot traps occurring during the daytime with one broad peak at mid- to late afternoon. The seasonal flight activity of I. duplicatus as monitoredbypheromone-baitedwindow-slottrapsduring2007‐2008indicatedthatthreemajorflightpeaksoccurredinearly June, late June‐early July and late July respectively, suggesting the existence of a potential second generation. CONCLUSION:Theoptimaltrapcharacteristicswillimprovetheperformanceofpheromone-baitedtrapsasacriticalmonitoring or mass-trapping tool to combat outbreaks of this pest species. c � 2009 Society of Chemical Industry

We screened 150 olfactory sensilla in single-sensillum recordings to unravel the mechanisms underlying host selection in the spruce bark beetle, Ips typographus (Coleoptera: Curculionidae: Scolytinae). Odor stimuli comprised of pheromone (various bark beetle spp.), host, and non-host compounds elicited strong and selective responses from 106 olfactory receptor neurons (ORNs), 45 of which were tuned to pheromone compounds, 37 to host compounds, and 24 to non-host volatiles (NHV). In addition, 26 ORNs responded only weakly to any odor stimulus. Strongly responding ORNs were classified into 17 classes. Seven classes responded primarily to the Ips pheromone compounds: cis-verbenol, ipsenol, ipsdienol (two classes), 2-methyl-3-buten-2-ol, amitinol, or verbenone, respectively. Six classes responded to the host compounds: alpha-pinene, myrcene, p-cymene, myrcene and p-cymene, 1,8-cineole, or Delta3-carene, respectively. Four classes responded to NHV: 3-octanol, 1-octen-3-ol, trans-conophthorin, or indiscriminately to the repellent green leaf volatiles (GLVs) 1-hexanol, Z3-hexen-1-ol and E2-hexen-1-ol, respectively. Indiscriminate responses from GLV neurons might explain a behavioral redundancy among these GLVs. This is the first description of individual bark beetle ORNs dedicated to NHV perception. These comprise almost 25% of the strongly responding neurons, demonstrating that a large proportion of the olfactory system is devoted to signals from plants that the insect avoids.

Volatiles from Ips duplicatus male hindgut extracts and aeration samples of spruce logs colonized by the virgin males were analyzed by coupled gas chromatography-mass spectrometry (GC-MS), coupled gas chromatography-electroantennographic detection (GC-EAD), and field trapping bioassays in Inner Mongolia, China. GC-MS analyses showed that males from nuptial chambers (for reproduction) produced hindgut volatile profiles similar to those of maturation feeding males from the nonreproductive galleries. The known aggregation pheromone components, ipsdienol (Id) and E-myrcenol (EM) are the major constituents of this blend, followed by several minor bark beetle-related compounds: 2-methyl-3-buten-2-ol, ipsenol, cis-verbenol, verbenone, 2-phenylethanol, myrtenol, and trans-myrtanol. The enantiomeric composition of the Id from the male hindgut extracts was determined to be a racemic mixture (50/50) by enantioselective GC. GC-EAD experiments indicated that I. duplicatus antennae from both sexes responded strongly not only to the known aggregation pheromone components, Id and EM, but also to the other minor conspecific-produced compounds. The strongest antennal response was elicited by the major pheromone component, Id, which was ca. 2-3 times higher than that elicited by the second component, EM. Electroantennogram dose-responses indicated that antennal response threshold to Id was approximately 100 times lower than that of EM. No synergistic effects between these occurred at peripheral level. A strong and repeatable EAD response was found to a trace compound (flame ionization detection undetectable) from both hindgut and aeration samples. The compound was identified as amitinol (At) based on the retention time and a further EAD analysis of a synthetic mixture. In the field trapping experiments, At increased the attraction of I. duplicatus to the traps baited with a binary blend of EM/Id (1:1) in a synergistic manner when tested at a low release rate (1:1:0.1) but had no effect on catches at the high release rate (1:1:1). This result suggests that At might be a part of the Chinese I. duplicatus aggregation pheromone system. The production ratios of EM/Id from the Inner Mongolian population (EM/Id approximately 1:2.0-4) were twice as high as those reported from Europe (EM/Id approximately 1:9), corresponding well with the differences in the optimal response ratios.

Volatiles from hosts, non-hosts, interspecifics, and conspecifics of the Asian larch bark beetle, Ips subelongatus Motsch., were analyzed using both gas chromatographic-electroantennographic detection (GC-EAD) and gas chromatography/mass spectrometry (GC-MS) techniques, and field trapping bioassays in Inner Mongolia, China. GC-EAD experiments indicated that I. subelongatus antennae (both sexes) strongly responded not only to the major male-produced conspecific components, ipsenol, and ipsdienol, but also to other bark beetle compounds (cis-verbenol and verbenone), host monoterpenes (alpha-pinene, beta-pinene, and para-cymene) from Larix sp. logs, and non-host leaf (green leaf volatiles and geranyl acetone) and bark (C8-alcohols and trans-conophthorin) volatiles. Repeatable EAD responses were also found to two compounds from hindgut extracts that are undetectable by GC. One of these minor compounds was identified as amitinol. Field trapping experiments showed that the EAD-active, major male-hindgut component, racemic ipsenol, is the only individual compound that significantly attracted both sexes of I. subelongatus, whereas all other compounds, including previously reported pheromone components of European Ips cembrae, ipsdienol and 3-methyl-3-buten-1-ol, were unattractive. Ipsdienol, 3-methyl-3-buten-1-ol, or their binary blend had no synergistic or antagonistic effects on I. subelongatus attraction when combined with ipsenol, whereas cis-verbenol (a synomone) and verbenone (the antiaggregation semiochemical) inhibited its attraction to the ipsenol-containing attractive blend. A mixture of three EAD-active host monoterpenes, alpha-pinene, beta-pinene, and para-cymene, was unattractive, but interrupted the pheromone response of I. subelongatus. Geranyl acetone, one of the strong EAD-active non-host volatiles also significantly reduced the number of I. subelongatus captured in traps baited with ipsenol-containing attractive blend. Our results add support to the recent phylogenetic finding that European and Asian larch bark beetles should be regarded as two distinct species: I. cembrae infecting larch in Europe and I. subelongatus infesting larch in Asia.

When searching for suitable hosts in flight, especially in mixed forests, conifer- inhabiting bark beetles will encounter not only suitable host trees and their odours, but also unsuitable hosts and nonhost trees. Rejection of these trees could be based on an imbalance of certain host characteristics and/or a negative response to some nonhost stimuli, such as nonhost volatiles (NHV). 2 Recent electrophysiological and behavioural studies clearly indicate that conifer- inhabiting bark beetles are not only able to recognize, but also to avoid, nonhost habitats or trees by olfactory means. Green leaf volatiles (GLV), especially C6-alcohols, from the leaves (and partly from bark) of nonhost angiosperm trees, may represent nonhost odour signals at the habitat level. Specific bark volatiles such as trans-conophthorin, C8-alcohols, and some aromatic compounds, may indicate nonhosts at the tree species level. Flying bark beetles are also capable of determining whether a possible host is unsuitable by reacting to signals from conspecifics or sympatric heterospecifics that indicate old or colonized host tree individuals. 3 Combined NHV signals in blends showed both redundancy and synergism in their inhibitory effects. The coexistence of redundancy and synergism in nega- tive NHV signals may indicate different functional levels (nonhost habitats, species, and unsuitable hosts) in the host selection process. Combinations of NHV and verbenone significantly reduced the number of mass attacked host trees or logs on several economically important species (e.g. Dendroctonus ponderosae, Ips typographus, and I. sexdentatus). 4 We suggest a semiochemical-diversity hypothesis, based on the inhibition by NHV of bark beetle host-location, which might partly explain the lower outbreak rates of forest insects in mixed forests. This 'semiochemical-diversity hypothesis' would provide new support to the general 'stability-diversity hypothesis'. 5 Natural selection appears to have caused conifer-inhabiting bark beetles to evolve several olfactory mechanisms for finding their hosts and avoiding unsuit- able hosts and nonhost species. NHV and unsuitable host signals have potential for use in protecting trees from attack. The use of these signals may be facilitated by the fact that their combination has an active inhibition radius of several metres in trap test, and by the observation of area effects for several trees near inhibitor soruces in tree protection experiments. Furthermore, incorpora- tion of negative signals (such as NHV and verbenone) and pheromone-based

Host and habitat selection in flight by conifer bark beetles is governed by inhibition of attraction by non-host volatiles (NHV), in addition to the well-known attraction to kairomones from host plants and to aggregation pheromones from conspecifics. Antennally-active NHV from angiosperm birch and aspen trees were field tested on the European spruce bark beetle, Ips typographus (L.), by pheromone baited traps in Sweden using release rates of NHV comparable to those from a non-host tree. Trap catches were significantly reduced by 50-70% by individual NHV: trans-conophthorin, 3-octanol and 1-octen-3-ol from non-host bark, 1-hexanol and (Z)-3-hexen-1-ol from both leaves and bark and (E)-2-hexen-1-ol from leaves. Combined NHV signals in binary and ternary blends with the same overall release rates (ca 5 mg/day) showed both redundancy and synergism in their inhibitory effects. The redundancy occurred among individual green leaf alcohols (GLV-alcohols or C 6 -alcohols), C 8 -alcohols, and between these two groups, while trans-conophthorin and verbenone (Vn, from unsuitable host trees) showed significant synergistic effects between each other and with GLV-alcohols or C 8 -alcohols. The coexistence of redundancy and synergism in non-host chemical signals may indicate different functional levels (non-host habitats, species, and unsuitable host trees) of these negative volatile signals in the host selection process of conifer bark beetles. We showed an active inhibitory range (AIR) of a NHV-blend plus Vn on a standard pheromone bait to be at least 2 m, by using a central pheromone trap and surrounding concentric barrier trap rings with radii of 1, 2, and 4 m. The influx of bark beetles flying across concentric rings within the AIR was reduced by 55-99%, depending on the distance to the central pipe trap. Our findings support the hypothesis that as mixed forests have greater semiochemical diversity than pure host stands; they disturb olfactory guided host choice, and may reduce the possibility of outbreaks of conifer-infesting bark beetles. The synergistic effects as well as the active inhibitory range found in this study also suggest that optimal combinations of these NHV and verbenone may have potential in protecting forests against I. tyographus by reducing or stopping attacks on suitable host trees. Mixed stands with higher semio-chemical diversity disturb olfactory guided host choice and reduce the risk for outbreaks of specialist herbivores. This semiochemical-diversity hypothesis' provides new support to the general 'stability-diversity hypothesis'.

The experimental testing of the hypothesis of population reduction by mass trapping with the establishment of treatment/control areas is a logistic problem on landscape scale. We suggest a possible test based on time-series analysis of tree-mortality data. Our example is helped by: an island effect, a 20-year tree-mortality record, and a very competitive synthetic aggregation pheromone. The successful development of synthetic pheromone (ipsdienol and E-myrcenol) dispensers, and the dose-response of wick-dispensers is briefly described. In the 2000 ha of spruce (Picea mongolica) forest reserve at Baiyinaobao, surrounded by grassland, 80 traps were employed for 3 years, catching 0.5–1.7 million beetles/year. In spite of intensive conventional management since 1982 (17 years), the long-term tree mortality had been oscillating around 600. During the first three years of mass trapping, tree mortality was down to 118, 100, and 88, respectively. Neither natural population cycles nor weather conditions could explain the 3-year mortality drop. Cycles did not drive the drop, as auto-correlation function (ACF) and partial ACF plots do not show any clear periodicity, only a weak 5-year cycle is visible. Weather did not explain the drop, as the 3 years of mass trapping had average rainfall and higher than average temperatures. Climate data show negative correlations of tree mortality and temperatures on year basis and a 20-year trend of increased monthly averages, especially for July temperature. The increased temperature (global warming) will increase stress on trees, but mass trapping may have counterbalanced this new threat to the forest.

Non-host volatiles (NHV) were tested on antennae of Ips sexdentatus with coupled gas chromatographic-electroantennographic detection (GC-EAD). Significant antennal responses were found to aggregation pheromone components, ipsdienol and amitinol, an attraction inhibitor, verbenone, and non-host bark volatiles extracted from Betula pendula, including trans-conophthorin (tC) and alcohols. The tC, a blend of three green leaf alcohols (C6-alcohols), and a blend of two bark alcohols (C8-alcohols) were tested in combination for their ability to disrupt the attraction of the stenographer bark beetle to pheromone-baited traps, unbaited and pheromone-baited maritime pine logs. The strongest disruptive effect resulted from the addition of the tC combined with the blends of NHV alcohols at high release rate, achieving an 83% reduction in trap catches, 89% reduction of attacks on unbaited pine logs, and 60% on baited logs. It is proposed that the two types of NHV have an additive effect, tC playing a major role in pheromone disruption while the alcohols would interrupt host specific signals. The results suggest that I. sexdentatus uses both bark and green leaf volatiles from non-host tree species in the host selection process. These NHV are promising disruptants, which could be used to prevent stenographer bark beetle attacks on log piles or standing trees in forest. A mixture of conifers and broad-leaved species would also disturb olfactory guided host selection supporting the assumption that more diverse forests are less prone to pest insect outbreaks.

Pheromone communication systems have a reliable signal with a restricted window of amounts and ratios released and perceived. We propose a model based on a Gaussian response profile that allows a quantification of the response peak (location of optimum) and a measure of the peak width (response window). Interpreting the Gaussian curve, fitted by nonlinear regression (NLR), as a standard normal distribution, the peak location equals the mean (μ) and the window width equals 2× the standard deviation (2σ). The NLR procedure can provide an objective measure for both peak location and width for a wide range of data sets. Four empirical data sets as well as 10 literature data sets were analyzed. The double-spined spruce engraver, Ips duplicatus, was field tested in four populations to find the optimum proportion for attraction to the two male aggregation pheromone components, ipsdienol (Id) and (E)-myrcenol (EM), ranging from 0 to 100% of Id. Tests in Norway and the Czech Republic confirmed the preference of western populations for a blend between 50 and 90% Id. A population in Inner Mongolia showed a preference for traps with the 10 and 50% Id baits. The NLR fitted values for response peak and width (μ; 2σ) were: Norway 0.64, 0.73; Czech Republic 0.53, 0.73; NE China 0.77, 0.29; and Inner Mongolia 0.33, 0.50. The signal produced by Norwegian field-collected males had a narrower window width (2σ = 0.12). Males of the maize stem borer, Chilo partellus, were tested in a flight tunnel for their response to variation in the two major female sex pheromone gland components, (Z)-11-hexadecenal and the corresponding alcohol (OH). Variation of the alcohol in seven levels from 2 to 29% OH showed the highest male response for 17% OH. For all behavioral steps, the peak of male response was near μ = 0.14, while the window width fell from 2σ = 0.5 to 0.2 for eight sequential behavioral steps from take-off to copulation. Female production had a similar peak location (μ = 0.13) but a narrower width, 2σ = 0.14. Literature data from other moth species showed similar patterns, with a wider male response relative to the female production windows. Literature data on response to enantiomer ratios in a hymenopteran and to pheromone amounts in a dipteran were also described by our model. In a bark beetle population (Ips pini), with two hybridizing enantiomeric strains, the production peaks were narrower (0.1) than the response peaks (0.5). Thus, it in general, seems that in the pheromone systems analyzed, the width of the response window (2σ = 0.1 to 0.8) is larger than that of the production window (2σ = 0.03 to 0.14), irrespective of the sex of the sender.

Volatiles from bark of three nonhost angiosperm trees of Ips typographus (Coleoptera: Scolytidae) (Betula pendula, B. pubescens, and Populus tremula) were collected by headspace sampling and direct solvent extraction in June 1998, and identified and quantified by coupled gas chromatographic-mass spectrometry (GC-MS). Only small amounts of bark volatiles were detected in the aerations in situ from undamaged stems of the nonhost trees. In headspace volatiles from bark chips the two birch species had a similar odour profile, with two sesquiterpene hydrocarbons, α-zingiberene and α-santalene, as the most dominant components. Bark of P. tremula emitted significantly less sesquiterpenes and more green leaf volatiles (GLVs) than the two birch species did. Solvent extraction of fresh bark gave similar volatile composition for the two birches compared to the headspace of bark chips, but large differences were found in P. tremula. Coupled gas chromatographic-electroantennographic detection analysis (GC-EAD) of the headspace volatiles from fresh bark chips of the three nonhost species revealed five compounds that consistently elicited antennal responses by I. typographus. The strongest antennal responses were elicited by trans-conophthorin, (5S,7S)-(–)-7-methyl-1,6-dioxaspiro[4.5]decane with optical purity of ca. 90% (S,S). The other four antennally active bark volatiles were 1-hexanol, (Z)-3-hexen-1-ol, 3-octanol and 1-octen-3-ol, which had similar electroantennogram (EAG) dose-response curves and response thresholds. (±)-trans-Conophthorin showed a different, linear EAG dose-response curve, with a 10 times lower response threshold than the other GC-EAD active compounds, similar to the pheromone component, (−)-(4S)-cis-verbenol. These results suggest that not only the green leaf alcohols from leaves of the angiosperm trees but also the nonhost bark volatiles could be used by I. typographus as long-range olfactory cues to discriminate between its conifer host and the nonhosts, birch and aspen.

Synthetic pheromone trap captures of Hyphantria cunea (Drury) (Lep., Arctiidae) males were compared with those of virgin female-baited traps and with visual observations. Delta-traps with females and Uni-traps with synthetic pheromone (1Z,3Z,6Z-9,10-epoxyheneicosatriene, 3Z,6Z-9,10-epoxyheneicosadiene, and 9Z,12Z,15Z-octadecatrienal; total, 2–6 mg/dispenser) and visual observation at a fixed point were used. The diurnal rhythm of activity in the little studied first (spring) generation showed a broad peak (3 h) after sunset. The second (summer) generation showed a short (1.5 h) but intense peak of attraction before sunrise, both to natural and to synthetic pheromones. A second small peak, just after sunset, was detected only in traps with the synthetic bait. Males started to be attracted to females about 1 h later than to synthetics in both generations. The main flight of males (mid-50% of cumulative male catches) was longer (8–11 days) in the first than in the second generation (5–7 days). The catches of males were increased by high daily maximum temperatures in the spring generation, while rain suppressed activity (phenology was delayed) in both generations. The total period of activity detectable by visual observations was about 6 days shorter than by pheromone traps. Population trends were recorded over four generations 1992–94 in two sites by trapping of moths and counting of larval webs. The second (summer) generation was two to six times larger than the spring generation. There was a correlation between the log-transformed catch of males and larval webs (r = 0.92 for proportions of both infested trees and web density) of the same period. This allows prediction of the subsequent larval abundance (offspring) based on adult (parent) pheromone trapping in preceding weeks.

Single-cell recordings from olfactory sensory neurons (OSNs), housed in sensilla located at the base and at the tip of the antenna, showed selective responses to plant odors and female sex pheromone in this polyphagous moth. A spatial variation existed in sensitivity: OSNs present on the more proximal segment (P) were more sensitive than those on the more distal segment (D). OSNs of the 2 locations also differed in temporal characteristics: OSNs on P had shorter latency and displayed more phasic responses, whereas those on D had more tonic responses, especially at low stimulus concentrations. The 196 OSNs responding to our 35 monomolecular stimuli in the screening were housed in 32 functional sensillum types: 27 in basiconic, 3 in long-trichoid, 2 in coeloconic, and 3 in auricillic sensilla. The OSNs in basiconic, coeloconic, and auricillic sensilla responded to plant-associated odorants, whereas OSNs in long-trichoid sensilla responded to female-produced sex pheromone components. Short-trichoid sensilla showed spontaneous activity, but no responses to any odorant tested. OSN specificity to plant stimuli ranged from highly specific to broadly tuned, but it did not differ clearly from females in more specialized moths. OSN response diversity is discussed in terms of olfactory coding, behavior, and ecological specialization.

Volatiles from bark of aspen,Populus tremulaL. and two species of birch: silver birch (Betula pendulaRoth.) and common birch (B. pubescensEhrh.), were collected by direct solvent extraction and aeration of both newly cut bark chips and undamaged stems in June 1998 and subjected to GC-MS analysis. The results showed the presence of 2-methyl-3-buten-2-ol (MB), one of the two principal aggregation pheromone components of the spruce bark beetle,Ips typographus, in bark extraction samples of all the three deciduous tree species tested. In addition, one more oxygenated hemiterpene, 3-methyl-3-buten-2-one, and (E)-3-penten-2-ol were also found in the bark extracts. Only trace amounts of MB were detected in some aeration samples of the fresh bark chips, and no MB was found from the aeration samples of undamaged stems at detectable levels. The occurrence of this compound was also confirmed in the bark of four exotic birch species:B. albosinensisSchneid.,B. ermaniiCham.,B. jacquemontiiSpach, andB. maximowiczianaRegel, but not yet in the European pines/spruces and the common yeasts. Our results raise major questions regarding the evolution, the tropospheric chemistry, and the ecological role of this hemiterpene alcohol. They also suggest that comparative studies on the biosynthetic pathways for MB in different sources would be of considerably evolutionary interest.

The biosynthesis of ipsdienol and E -myrcenol was studied in the bark beetle Ips duplicatus by using the enzyme inhibitor compactin. The inhibitor specifically blocks the action of the enzyme hydroxy-methylglutaryl-coenzyme A reductase, that operates early in terpene biosynthesis prior to the formation of mevalonate. Compactin was injected in the beetle's abdomen using a thin tapered glass needle; controls were injected with saline blank. The release of ipsdienol and E -myrcenol was reduced by 70 and 40% respectively in I. duplicatus beetles injected with compactin over a period of 17–25 h after injection. Thus, ipsdienol and E -myrcenol can be synthesized de novo and not only by the hydroxylation of exogenous myrcene, from the resin of the attacked host trees. To test the possibility that the inhibitor causes a non-specific metabolic depression of the beetles, I. typographus was subjected similarly and monitored for the hydroxylation of (−)-α-pinene, forming cis -verbenol. The production of cis -verbenol is not affected by compactin.

We identified several compounds, by gas chromatographic–electroantennographic detection (GC–EAD), that were antennally active in the bark beetle Ips typographus and also abundant in beetle-attacked spruce trees. One of them, 1,8-cineole (Ci), strongly inhibited the attraction to pheromone in the field. Single-sensillum recordings (SSRs) previously showed olfactory receptor neurons (ORNs) on I. typographus antennae selectively responding to Ci. All Ci neurons were found within sensilla co-inhabited by a pheromone neuron responding to cis-verbenol (cV); however, in other sensilla, the cV neuron was paired with a neuron not responding to any test odorant. We hypothesized that the colocalization of ORNs had a functional and ecological relevance. We show by SSR that Ci inhibited spontaneous activity of the cV neuron only in sensilla in which the Ci neuron was also present. Using mixtures of cV and Ci, we further show that responses to low doses (1–10 ng) of cV were significantly reduced when the colocalized Ci neuron simultaneously responded to high doses (1–10 μg) of Ci. This indicated that the response of the Ci neuron, rather than ligand–receptor interactions in the cV neuron, caused the inhibition. Moreover, cV neurons paired with Ci neurons were more sensitive to cV alone than the ones paired with the non-responding ORN. Our observations question the traditional view that ORNs within a sensillum function as independent units. The colocalization of ORNs might sharpen adaptive responses to blends of semiochemicals with different ecological significance in the olfactory landscape. * Ci : (±)-1,8-cineole cV : (4 S )- cis -verbenol GC–EAD : gas chromatographic–electroantennographic detection GC–MS : gas chromatography–mass spectrometry NHV : non-host volatile OBP : odorant-binding protein ORN : olfactory receptor neuron SSR : single-sensillum recordings

Ips duplicatus withI. typographus co-inhabiting Norway spruce (Picea abies (L.) Karst.) would benefit from a pheromone blend distinct from that of the larger competitorI. typographus. GC-MS analysis showed thatI. duplicatus males feeding in the host produced ipsdienol (Id),cis-verbenol (cV),trans-verbenol (tV), myrtenol (Mt), andE-myrcenol (EM) and traces of 2-methyl-3-buten-2-ol (MB).I. duplicatus produced Id in approximately racemic form (48.9-54.5% (+)-(S)-isomer). The amounts of Id and EM released over a 9 day period had a maximum of 250 and 5 ng/h/male, respectively, on day 2. Exposure ofI. duplicatus males to myrcene and α-pinene resulted in the production of small amounts of Id, cV, tV, Mt, andtrans-pinocarveol, but not of EM. In laboratory bioassays with walking beetles, the pheromone component Id alone was weakly attractive while EM was inactive, but in binary combination with Id strongly synergized attraction. A combination of EM and Id at a release rate equivalent to 100–200 males was more attractive in the field than 70 unmated males in a spruce log. The addition of myrcene ( a suggested pheromone precursor of Id) to Id did not enhance trap caches, while addition of EM increased catches > 10-fold. Subtracting EM from a blend of Id, EM, cV and MB drastically reduced trap catches while subtraction of cV or MB or both had no significant effect. Addition of EM over a wide concentration range to the synthetic pheromone ofI. typographus did not reduce the attraction of females of this species in the laboratory. A two-species pheromone interaction field test releasingI. typographus pheromone components (MB + cV) at 10–1000 male equivalents (ME) andI. duplicatus pheromone (Id + EM) at 0, 10–1000 ME in all possible combinations showed both positive intraspecific dose-response effects and an interspecific inhibition. Higher release rates of EM appeared to inhibitI. typographus, especially males. In a tree colonization model, the response of the two competing species to their respective pheromones show a good separation during the mass-attack with a small initial cross-attraction. It remains to be shown whether either of the two pheromone systems have in fact evolved in the present sympatry, or if they are an incidental effect of ancestry of these phylogenetically distantIps.

There is a need for information on communication distances in natural systems and the ranges of action of synthetic pheromone traps. The differences between sampling range (R S , time dependent ecological measure) and attraction range (R A , behavioural measure) are stressed and compared to the simple but artificial effective attraction radius concept (R EA , mainly trap and bait dependent). To estimate such ranges defined theoretical concepts and proper analysis of data are needed (.)

Background Insect predators and parasitoids exploit attractive chemical signals from lower trophic levels as kairomones to locate their herbivore prey and hosts. We hypothesized that specific chemical cues from prey non-hosts and non-habitats, which are not part of the trophic chain, are also recognized by predators and would inhibit attraction to the host/prey kairomone signals. To test our hypothesis, we studied the olfactory physiology and behavior of a predaceous beetle, Thanasimus formicarius (L.) (Coleoptera: Cleridae), in relation to specific angiosperm plant volatiles, which are non-host volatiles (NHV) for its conifer-feeding bark beetle prey. Methodology/Principal Findings Olfactory detection in the clerid was confirmed by gas chromatography coupled to electroantennographic detection (GC-EAD) for a subset of NHV components. Among NHV, we identified two strongly antennally active molecules, 3-octanol and 1-octen-3-ol. We tested the potential inhibition of the combination of these two NHV on the walking and flight responses of the clerid to known kairomonal attractants such as synthetic mixtures of bark beetle (Ips spp.) aggregation pheromone components (cis-verbenol, ipsdienol, and E-myrcenol) combined with conifer (Picea and Pinus spp.) monoterpenes (α-pinene, terpinolene, and Δ3-carene). There was a strong inhibitory effect, both in the laboratory (effect size d = −3.2, walking bioassay) and in the field (d = −1.0, flight trapping). This is the first report of combining antennal detection (GC-EAD) and behavioral responses to identify semiochemical molecules that bypass the trophic system, signaling habitat information rather than food related information. Conclusions/Significance Our results, along with recent reports on hymenopteran parasitoids and coleopteran predators, suggest that some NHV chemicals for herbivores are part of specific behavioral signals for the higher trophic level and not part of a background noise. Such bypass-trophic signals could be of general importance for third trophic level players in avoiding unsuitable habitats with non-host plants of their prey.

Adults of the sorghum chafer, Pachnoda interrupta Olivier (Coleoptera: Scarabaeidae: Cetoniinae), form aggregations during the mating period in July, but also in October. The beetles aggregate on food sources, e.g., Acacia spp. trees or sorghum with ripe seeds, to feed and mate. During the mating season, field trapping experiments with live beetles as bait demonstrated attraction of males to unmated females, but not to mated females or males, indicating the presence of a female-emitted sex pheromone. Unmated females combined with banana (food source) attracted significantly more males and females than did unmated females alone. Other combinations of beetles with banana were not more attractive than banana alone. Thus, aggregation behavior appears to be guided by a combination of pheromone and host volatiles. Females and males were extracted with hexane during the mating period, and the extracts were compared by using GC-MS. In a field trapping experiment, 19 compounds found only in females were tested, both singly and in a mixture. Traps baited with one of the female-associated compounds, phenylacetaldehyde, caught significantly more beetles than any other treatment. However, the sex ratio of beetles caught in these traps did not differ from that of control traps, and it is possible that other components may be involved in the sex pheromone signal. Furthermore, traps baited with a mixture of all 19 compounds attracted significantly fewer beetles than did phenylacetaldehyde alone.

Males of the Eurasian bark beetleIps duplicatus, when feeding in host Norway spruce (Picea abies (L.) Karst.), produced and released ipsdienol andE-myrcenol, which we show to be aggregation pheromone components. Bioassays using walking beetles indicated thatE-myrcenol in synergistic combination with ipsdienol is essential for attraction. Synergism ofE-myrcenol and ipsdienol released at natural rates in the forest was also demonstrated with a new technique using mechanical slow-rotation of sticky traps.

Newly emerged males ofHyphantria cunea (Drury) (Lepidoptera: Arctiidae), marked with different fluorescent colors, were recaptured by pheromone trapping. Three Uni-traps, baited with (3Z,6Z)9,10-epoxyheneicosadiene, (9Z,12Z,15Z)octadecatrienal, and (3Z,6Z)9,10-epoxyheneicosa-triene in a 1:10:1 ratio, total 6 mg/dispenser, were placed in a line 15 m apart, perpendicular to the dominant wind direction, and checked after 12, 36, and 60 hr. At dusk, releases of 10-25 males were made over distances from 30 to 250 m, during 4 periods over a soybean field (nonhost plant). Recapture rates were high; of a total of 176 males released, 115 were recaptured with 88% of these within 12 hours. Between 40 and 100% of males were recovered over shorter distances (30-150 m), and between 10-24% at longer distances (200 and 250 m). No other species or unmarked males were captured. The trapping period showed little effect on recaptures. The central trap of the three traps had somewhat higher catches, but the slope of the regression of recapture over distance did not differ among traps. There was a significant decline of recapture over distance (r=-0.56) for catches of individual traps but a large scatter. Summed recaptures per distance gave less scatter and hadr=-0.86, allowing calculation of the maximum sampling range (R s ) after 60 hr as 340 m with a 95% confidence interval of 190-710 m (regression of arcsin √p and √R). The area of sampling, found by integrating the probability function of recapture over distance, from the source to maximumR, was 7 ha. The high recapture rates and the longR,. compared to those in the literature for other taxa, indicate that pheromone traps are highly efficient sampling devices in this species.

A summary of the present knowledge of variation in individual quality within a bark beetle population is given, with emphasis on the spruce bark beetle Ips typogrophus, A major causal factor is density, mediated by competition during larval development. Density negatively influences individual quality measured as weight, fat content and pheromone production. Together with decreasing mean values at higher densities, the skewness of the frequency distributions goes from negative to positive, while variance changes little. High densities, which often occur in the field, thus result in a large fraction of “low quality beetles”. They have lower reproductive capacity and presumably lesser dispersal ability, lower survival, and earlier response to pheromone. This might concentrate the population in the next generation with increased competition as a result. It is suggested that a decrease in beetle “quality” due to increasingly intense intraspecific competition can contribute to the decline of an epidemic population.

A model of attack dynamics on a single tree under mass attack by the spruce bark beetle Ips typographus is developed from earlier studies of pheromone production and response of the flying population to the pheromone signal. The model is explored for different numbers and sex-ratios of pioneer beetles and responding beetles. Both numbers of pioneers and responders were important for the dynamics, while the sex-ratio of pioneers was less important. A decreased proportion of males among the responding beetles was important for the result of the model. In the early stage of the attack the model predictions of both numbers and sex-ratio of beetles landing were similar to examples from naturally mass-attacked trees, although the attack proceeded more slowly than in the field. Several aspects of the attack dynamics, such as density-regulation of the number of attacks on the tree, as observed in the field, were not predicted by the model. Gaps in the knowledge of the chemical ecology of the tree-insect relation, such as qualitative change in odours from the tree, spacing mechanisms on the bark surface, and mortality due to host resistence, are pointed out with help of the model.

A Y-tube olfactometer is described which was used to assay the response of walking Ips typographus (L.) towards the pheromone from beetles excavating galleries in logs of Norway spruce [Picea abies (L.) Karst]. Beetles in the logs were categorized into seven sequential attack phases. Males in the early attack phases were attractive as early as 0.5 to 2.5 h after their introduction. These logs remained attractive for more than 2 weeks. Walking females strongly preferred logs with unmated males over uninfested logs and over logs containing males and females. They also preferred logs with males and females over uninfested logs. Responding males showed a weaker attraction than females to logs with unmated males. The results are interpreted as adaptive responses of the sexes to variation of pheromone release from beetles in the host during subsequent phases of attack and colonization. Resume Marche d'Ips typographus L. en reponse aux pheromones naturelles produites pendant les differentes phases de contamination de l'epicea Un olfactometre a tube en Y, dont la description est donnee, a ete utilise pour examiner les reactions d'I. typographus L. par la marche a la presence de pheromones de congeneres creusant des galeries dans des buches d'Epicea (Picea abies L.). Les insectes dans les buches ont ete repartis en 7 classes sequentielles, depuis l'elaboration de la chambre nuptiale par le mâle (phases 1–3), l'accouplement (4), et la ponte jusqu'a ce que les galeries de ponte aient 5 cm (5–7). Les mâles dans les phases precoces (1–2) etaient attractifs pour les 2 sexes des 0,5 a 2,5 h apres leur introduction, et ils le restaient pendant plus de 2 semaines. Les femelles qui se deplacent par la marche ont prefere nettement les buches avec des mâles vierges a des buches non habitees. De meme elles ont prefere des buches avec des mâles unis aux femelles a des buches sans Ips. Ayant a choisir entre des buches infestees par des mâles vierges ou experimentes, les femelles ont prefere nettement les mâles vierges. Les mâles ont presente une plus faible attraction aussi bien pour les buches avec mâles vierges qu'avec mâles experimentes, ils n'ont pas manifeste de preference entre les 2. Les resultats sont interpretes comme des reponses adaptatives des 2 sexes aux variations dans la liberation de pheromones par les insectes pendant les phases successives d'attaque et de colonisation de l'hote.

The total amounts of, and proportions among, components of the aggregation pheromone produced byIps typographus were found to vary considerably among individuals excised from attacks on standing spruce trees. Chemical analyses of 392 individual male beetles were made by GC-MS. Both unmated and mated males had log-normal frequency distributions in their content of the pheromone components 2-methyl-3-buten-2-ol (MB) andcis-verbenol (cV), since a large fraction of males had a low content. The amount of MB in male hindguts varied independently of cV and the other oxygenated monoterpenes, while the amount of cV covaried with the other pinene alcohols and showed a variation between beetles from different spruce trees. Mated males had, on average, lower amounts of MB than unmated, while the average content of cV in mated males varied with the resin content of their host trees. Ipsdienol and ipsenol were only found in mated males, but in less than 40% and 10%, respectively, of these mated males. Even-aged males exposed to α-pinene in the laboratory showed slightly less variation in the amounts of verbenols, and the variations in ratio between cV and tV were similar to those among males attacking the same spruce tree.

The presently published data of variation in pheromone content in bark beetles (Coleoptera: Scolytidae) and moths (Lepidoptera) are reviewed. In both taxa high coefficients of variation, often up to around 100% for amount of pheromone, seem to be the rule. The contribution to the variation from errors in the chemical methods was small in most studies compared with the intrinsic biological variation. Examples arc given of variation in both absolute amounts, proportions of geometric isomers and in enantiomeric composition. Ratios of isomers often had lower coefficients of variation (25% or less). A lower average variation among sex pheromone in moths could relate to a mating system involving singly calling females, compared with the aggregation pheromone of bark beetles, which often call in large groups. The origin of a resource-based aggregation pheromone system by evolution working via individual selection is outlined.

The functions of the two synergistic pheromone compon- ents, (4s)-cis-verbenol (cV) and 2-methyl-3-buten-2-01 (MB), and the role of ipsdienol in the attraction of Ips typographus (L.) (Scolytidae) to pheromone sources were studied in the field. Absolute and relative beetle catches were compared between several traps placed at and nearby a central pheromone source: a pipe trap containing the source, a surround- ing sticky trap, a nearby window trap, and four distant window traps. A higher catch in the outer down-wind distant traps indicated an up-wind anemotaxis to the source. Increased MB release, with cV constant, increased the proportion caught in the central pipe trap, indicating MB as a landing stimulus. Release of MB alone gave a very small catch. Ipsdienol could not substitute for cV in the synergism with MB. An increase of cV, with MB constant, increased the number of beetles caught, but not the proportion caught in the pipe trap. The sex ratio was equal in the window traps, but fell to 30% males in both sticky and pipe traps, showing that a large proportion of the males attracted to the source did not land. The proportion of males in the pipe trap was reduced at the highest cV dose. The results support the idea of each pheromone component having a different relative importance in releasing different steps in the behavioural chain.

Six compounds previously identified from hindguts of unmated maleIps typographus (L.) during host colonization: 2-methyl-3-buten-2-ol (MB),cis-verbenol (cV),trans-verbenol (tV), myrtenol (Mt),trans-myrtanol (tM), and 2-phenylethanol (PE), were tested for their attractivity in the field with a subtractive method. The amounts of MB and cV released from a pipe trap were similar to those given off from the commercial bait Ipslure as well as that from a Norway spruce tree,Picea abies (L.) Karst., under mass attack. The blend of the compounds became nonattractive when either MB or cV was subtracted, while subtraction of any of the other four compounds had no effect. Addition of ipsdienol (Id) to the blend did not significantly increase the attraction. In a second comparative test, the addition of three compounds as a group (tV + Mt + PE) to MB + cV again had no effect on the attraction, but the addition of Id increased the catch somewhat. Addition of host logs to a bait releasing MB + cV at a rate lower than in previous experiments did not influence the attraction to pipe traps. Sticky traps containing natural pheromone sources (50 males in a log), which released 1-5 mg/day of MB as determined by aerations with deuterated MB as internal standard, were less attractive than a synthetic source releasing similar amounts of MB.

Ips typographus beetles were collected in the field, separated into eight attack phases (from beetles walking on the trunk of a tree under attack to those excavating gallery systems with a mother gallery longer than 4 cm), and immediately frozen in liquid nitrogen. 2-Methyl-3-buten-2-ol,cis- andtrans-verbenol, verbenone, myrtenol, trans-myrtanol, ipsenol, ipsdienol, and 2-phenylethanol were quantified from excised hindguts against an internal standard, heptyl acetate, in the extraction solvent. Methylbutenol, the pinene alcohols, and 2-phenylethanol showed the same pattern of variation between attack phases in males, with the largest amounts present before accepting females and then a fast decline. Ipsenol and ipsdienol were not detected in males before the females were accepted, and the amounts increased when the females start their egg laying. Verbenone occurred only in trace amounts. The beetles were sampled from five Norway spruce trees (Picea abies) of differing resin flow. The correlations between the nine pheromone components and five major host monoterpenes in the gut showed that the variation in the amount of methyl-butenol, ipsenol, and ipsdienol could not be explained by the variation in the amounts of host monoterpenes. In contrast over 80% of the quantitative variation ofcis-verbenol,trans-verbenol, and myrtenol was explained by the amount of α-pinene. The nine pheromone components from 36 individual males were also quantified. Both methylbutenol andcis-verbenol showed a large variation in both amounts and proportions. Females containedtrans-verbenol and traces of most other components found in males. When accepted by the male, they also contained a female-specific compound, β-isophorone. Behavioral and biosynthetic implications of the results are discussed.

The semiochemicals verbenone (Vn), ipsenol (Ie), and ipsdienol (Id), present in late phases of host colonization, have been implicated as qualitative “shut-off” signals regulating attack density. Combinations of the three chemicals were released in pipe traps together with the aggregation pheromone components 2-methyl-3-buten-2-ol (MB) andcis-verbenol (cV) at different levels and in different ratios to MB + cV, and with two spacings of traps to test for possible effects on reducing catch at traps baited with aggregation pheromone. When they were released with the attractants Vn and Ie (alone or together) decreased the mean catch significantly at the higher release rates used (1 mg/day). Id alone or together with Vn at low release rates (0.1 mg/day), with the attractants, increased catch somewhat. A dose-response test of Vn, with the attractants held constant, showed a decline in catches, down to about < 10% of the control, at ratios of Vn to cV between 1∶1 and 150∶ 1. A larger spacing (25 m) of traps gave a stronger response to change in doses of Vn and MB + cV than a smaller (6 m) spacing. The sex ratio was more skewed towards females when two or three inhibitors were present and at higher doses of Vn. It is suggested that Vn could be the most important density-regulating signal in the natural system, as release of Vn from galleries is larger and starts earlier than that of Id and Ie.

The density of bark-beetle colonization of a tree could be regulated by a quantitative effect of the pheromone signal from beetles in the tree (cessation of release of attractive pheromone) or by a qualitative effect (production of pheromone components inhibiting attraction). The quantitative hypothesis was tested onIps typographus by varying the release rate of the two known attractive compounds, 2-methyl-3-buten-2-ol (MB) and (4S)-cis-verbenol (cV). The highest number of beetles were captured at traps with the highest release rates. The catch was nearly proportional to the release of MB and cV at a distance between traps of 12 m or more. At 6-, 3-, and 1.5-m distances between traps deployed in a triangular arrangement there was still a good discrimination between release rates, but relatively more beetles, especially males, were caught on the blank. The lower release rates caught an equal sex ratio while the highest release rate caught only about 30% males. The qualitative hypothesis was tested by releasing the suspected inhibitors ipsdienol (Id) and ipsenol (Ie), from traps in the same amounts as cV. Only small effects were noted forI. typographus. However, the competitorI. duplicatus was attracted to Id and inhibited by Ie, while the predatorThanasimus formicarius was attracted to both compounds. On the other hand, when the ratio of Id or Ie to cV was 10∶1 or 0.1∶1 rather than 1∶1, they affected the numbers ofI. typographus attracted. A small amount of Id combined with the attractants increased trap catch, while large amounts of Id or Ie decreased attraction, especially when combined. Attack density regulation is modeled as an effect of both quantitative and qualitative mechanisms acting in sequence.

In the laboratory we investigated the effect of breeding density on the reemergence of parent spruce bark beetles, Ips typographus (L.), and on the number and "quality" of their offspring. The parents reemerged sooner at higher densities but the total proportion that reemerged was independent of density. More than 20 offspring per female were produced at the lowest density (0.5 99 /100 cm2)but only 0.6 per female at the highest density (31 99/100 cm2). The offspring from the lowest density were about 50% heavier than those from the highest density and also the fat content (%) increased with decreasing density. Females weighed less and contained less fat than males. Male offspring from lower densities produced larger amounts of the pheromone components cis-verbenol and 2-methyl-3-buten-2-ol than males from the higher densities. Offspring from the highest density produced about half as many progeny as those from the lowest densities, showing an effect of density acting over more than one generation. These findings suggest that density-induced variation of beetle "quality" might be of importance in the population dynamics of bark beetles.