Volatile organic compounds and host-plant specialization in European corn borer E and Z pheromone races

Plant volatile cues are considered the main source of information for ovipositing moths, which use chemical information to locate and recognize the host plant. In Europe, two sympatric populations of European corn borer (ECB; Ostrinia nubilalis, Hübner), the Z and E-pheromone races, feed mainly on maize and hop or mugwort, respectively. We studied the mechanisms of host-plant recognition and fidelity in ECB pheromone races by testing the attractiveness of host plants to gravid females in a flight tunnel and by analyzing the volatiles released from maize, mugwort, and hop during the scotophase, when the ovipositing flight of the ECB females occurs. In the wind tunnel bioassay, the Z-race and E-race females engaged in upwind flight and expressed a strong host fidelity to their respective main host plants; all three of these host plants possess distinctive volatile profiles specific as to blend and ratio. The host plants shared a certain number of ubiquitous volatiles present in various ratios that likely constitute a species-specific cue to host-seeking ECB moths. Our observations therefore suggest that ECB host fidelity is steered by plant volatiles that are present in species-specific ratios of ubiquitous volatile organic compounds.     

The use of plant volatiles for host location by an ash (Fraxinus) specialist,Caloptilia fraxinella

Volatile organic chemicals (VOCs) are used by female moths to find host plants for oviposition and specialist moths can be highly tuned to the volatile signature of their host plant. The ash leaf-coneroller, Caloptilia fraxinella (Ely) (Lepidoptera: Gracillariidae) specializes on ash (Fraxinus spp.) (Oleaceae). Its introduction to urban forests in the Canadian Prairie Provinces on both green, F. pennsylvanica, and black ash, F. nigra, offers the opportunity to test odor-mediated host location to two host-plant species. In laboratory and field experiments, C. fraxinella adults oriented to volatiles released from ash seedlings. The antennae of mated female C. fraxinella consistently detected five VOCs released from black and green ash, four of which were common to both species. Blends of natural and synthetic VOCs found to elicit an antennal response were tested in wind tunnel and field bioassays. Synthetic and natural VOCs elicited as much oriented flight from mated female C. fraxinella as ash seedlings, but did not elicit contact with the VOC lure in the wind tunnel. In the field, traps baited with blends of synthetic copies of black and green ash VOCs did not attract more female C. fraxinella than unbaited control traps. These experiments lay the foundation for further research to develop semiochemical lures to attract female C. fraxinella.     

Floral scent variation in two <Emphasis Type="Italic">Antirrhinum majus</Emphasis> subspecies influences the choice of naïve bumblebees

Two wild subspecies of snapdragon, Antirrhinum majus, subspecies pseudomajus and striatum, differ in floral color and can be visually discriminated by insect visitors. The extent to which olfactory cues derived from floral scents contribute to discrimination between snapdragon subspecies is however unknown. We tested whether these two subspecies differ in floral scent and whether these olfactory differences are used by bumblebees (Bombus terrestris) to discriminate between them. We grew individuals of both subspecies, collected from a total of seven wild populations, under controlled conditions. We quantified the volatile organic compounds (VOCs) emitted by the flowers using gas-chromatography/mass-spectrometry/flame-ionization-detection. We studied antennal detection of VOCs by bumblebees, by means of electroantennogram study (EAG). We also performed behavioral experiments in a Y-maze to determine the innate response of bumblebees to the main floral VOCs emitted by our snapdragon subspecies. The floral scent of Antirrhinum majus pseudomajus contained three volatile benzenoids absent in the floral scent of Antirrhinum majus striatum. One of them, acetophenone, contributed over 69% of the absolute emissions of A. majus pseudomajus. These benzenoids elicited a significantly higher EAG response compared with other VOCs. In the Y-maze, bumblebees were significantly less attracted by acetophenone, suggesting an aversive effect of this VOC. Our findings indicate that bumblebees are able to discriminate between the two Antirrhinum majus subspecies. Differences in flower scent between these subspecies and olfactory bumblebee preferences are discussed in the light of biochemical constraints on VOCs synthesis and of the role of flower scent in the evolutionary ecology of A. majus.     

Differences in the electroantennal responses of apple- and hawthorn-infesting races of Rhagoletis pomonella to host fruit volatile compounds

Summary. Domestic apple (Malus pumila)- and hawthorn (Crataegus sp.)-infesting races of Rhagoletis pomonella, Walsh (Diptera: Tephritidae) provide an excellent model to examine the role that host plant specificity plays during sympatric speciation (i.e., divergence in the absence of geographic isolation). Previous work has shown that these races differ in their propensities to accept apple and hawthorn fruits in behavioral choice assays, and that this discrimination translates into "host fidelity" in the field (i.e., apple flies tend to mate on and oviposit into apples and hawthorn flies on hawthorns). ?We present the results of a study examining possible physiological factors contributing to host choice differences in R. pomonella. We tested whether apple and hawthorn flies differ in their electroantennogram (EAG) responses to biologically relevant volatile compounds emitted from apples and hawthorns. Significant differences were found in the relative EAG responses of apple and hawthorn flies to host fruit compounds at five of six paired study sites across the eastern United States. The geographic pattern of EAG variation was complex, however, with local populations of apple and hawthorn flies tending to be more similar to one another than to flies of the same race at distant sites. This pattern was largely due to EAG responses for several compounds showing longitudinal or latitudinal clines, the latitudinal clines being similar to those observed for allozyme loci in the host races. We also found evidence for sex-related differences, as males tended to have higher mean EAG responses to compounds than females. Host-associated differences were therefore nested within geographic and sex-related differentiation in R. pomonella.?Further behavioral studies are needed to distinguish whether the EAG differences are responsible for, as opposed to being a consequence of, host-plant fidelity and adaptation. Crosses are also required to establish a genetic basis for the EAG responses, although we did find significant correlations between EAG scores for several compounds and the allozymes NADH-Diaphorase-2 and Hydroxyacid dehydrogenase at one of the study sites. Questions therefore remain concerning the evolutionary significance of the EAG response differences between apple and hawthorn fly races. Nevertheless, these differences raise the possibility that antennal responses to fruit-related volatile compounds contribute to host plant discrimination in R. pomonella. Regardless, the EAG responses represent another set of traits, in addition to diapause/eclosion time phenotypes and allozyme frequencies, differing between apple and hawthorn host races of R. pomonella. Received 17 March 1998; accepted 21 September 1989.     

Electrophysiological responses of Dendroctonus armandi (Coleoptera: Curculionidae: Scolytinae) to volatiles of Chinese white pine as well as to pure enantiomers and racemates of some monoterpenes

To better understand the attractiveness of host tree, Chinese white pines (Pinus armandi Fr.) to Chinese white pine beetle (Dendroctonus armandi Tsai and Li), the antennal responses of D. armandi to the host volatile, as well as the pure enantiomers and racemates of some monoterpenes, were examined using an electroantennogram (EAG). EAG responses of male and female D. armandi to blended volatiles extracted from the host and some synthetic terpenes (α-phellandrene, (−)-β-pinene, (+)-α-pinene, (−)-α-pinene, (−)-camphene, β-myrcene, (S)-(−)-limonene, (+)-camphene and (R)-(+)-limonene) showed significant variation due to different compound concentrations and sex of the beetles. EAG responses to extracted blended volatiles were significantly greater in females than in males, but the EAG response was not always proportional to the volatile concentration. At lower concentrations, females responded strongly to α-phellandrene and males to (−)-β-pinene, while at higher concentrations, females responded most strongly to α-phellandrene and males to (+)-α-pinene. Females were significantly responsive to (−)-α-pinene, α-phellandrene and (−)-camphene, while males were more responsive to (S)-(−)-limonene, (+)-α-pinene and (R)-(+)-limonene. The EAG responses of the female D. armandi to the volatile oil were significantly higher than that of the males, and the infested pine volatiles could evoke higher EAG response. Most of the test compounds elicited similar responses, which suggested that several of the compounds may be used in combination by D. armandi in habitat and/or host community location at the Qinling forest ecosystem.     

Weak defence in a tritrophic system: olfactory response to salicylaldehyde reflects prey specialization of potter wasps

Predatory arthropods are attracted to infochemicals emitted by their herbivore prey or by the prey’s host plants. We studied such a tritrophic system measuring the olfactory responses of three potter wasp species (Symmorphus murarius, Symmorphus gracilis, Discoelius zonalis, Hymenoptera: Eumeninae) to salicylaldehyde, sequestered as a defence compound by Chrysomela leaf beetle larvae when feeding on Salicaceae, and volatile organic compounds (VOCs) emitted by aspen (Populus tremula, Salicaceae). In electroantennographic recordings (EAG), the highly specialized S. murarius that almost exclusively feeds on larvae of Salicaceae-feeding Chrysomela species was more sensitive to salicylaldehyde than the less specialized S. gracilis, feeding on such Chrysomela species but also weevil larvae. In contrast the related D. zonalis, foraging for microlepidoptera caterpillars on various host plants, did not respond at all. Furthermore, the three wasp species responded differently to aspen VOCs in GC–MS/EAD measurements. These results indicate that the sense of smell of predatory potter wasps differs for prey and plant volatiles among related wasp species according to their degree of host specialization. The considerable differences in salicylaldehyde perception suggest that its originally defensive function has backfired as it is used by specialist potter wasps for prey location. This is an important clue on adaptive mechanisms of the highest trophic level of the well-studied evolutionary arms race among Chrysomela leaf beetles, their host plants and their enemies.     

Antennal electrophysiological responses of the giant swallowtail butterfly, Papilio cresphontes, to the essential oils of Zanthoxylum clava-herculis and related plants

We used the electroantennogram (EAG) technique to compare the antennal sensitivity of both sexes of the giant swallowtail butterfly, Papilio cresphontes to four doses (1, 10, 100, and 1,000 μg) of the leaf essential oils of Zanthoxylum clava-herculis and Ptelea trifoliata (key host plants) and Sassafras albidum (a marginal or non-host plant). The main hypothesis tested was that P. cresphontes will show greater olfactory sensitivity to volatiles of the key host plants than to volatiles of the marginal host plant, in particular at low doses. At the lower doses, extract of the key host plant, Z. clava-herculis elicited greater EAG responses in both sexes than extracts of the remaining two plants. At higher doses, however, extracts of P. trifoliata and S. albidum elicited greater EAG responses than extract of Z. clava-herculis. These results partly support our hypothesis and may suggest that Z. clava-herculis is a more preferred host plant of P. cresphontes than P. trifoliata. In general, female butterflies showed greater EAG responses than males to the three plant extracts at the higher doses. Preliminary coupled gas chromatography-electroantennogram (GC-EAD) tests revealed four components each from Z. clava-herculis and P. trifoliata (three peaks common to both extracts) and seven from S. albidum (one shared with Z. clava-herculis) which elicited GC-EAD activity in P. cresphontes females, but the peaks were un-indentified because most were detected in trace amounts. In addition, the chemical composition of the leaf essential oil of Z. clava-herculis was analyzed by GC–MS. The leaf essential oils consisted of 25 components, largely menthane monoterpenoids, dominated by limonene and 1,8-cineole, but neither of the two major components elicited significant GC-EAD response in P. cresphontes. These results are discussed in relation to host-plant selection in P. cresphontes.     

Behavioral and electroantennographic responsiveness of adult butterflies of six nymphalid species to food-derived volatiles

While adults of most species of butterflies visit flowers for food (nectar), those of certain species can feed on rotting food (e.g., exuded tree sap and rotting fruits). These food sources considerably differ in odor; flowers emit species-specific scents and rotting-food odors are characterized by fermentation products. To determine whether adult butterflies have different olfactory performances adaptive to their food selections, we examined the following responses of naïve adults of six nymphalid species, namely, Nymphalis xanthomelas, Kaniska canace, Vanessa indica, Cynthia cardui, Hypolimnas bolina, and Argyreus hyperbius: the proboscis extension reflex (PER) was evaluated as the behavioral criterion of preference for scent compounds, and the electroantennographic (EAG) response was assessed as the sensory criterion of antennal perception. All of these species are nectarivorous, but N. xanthomelas, K. canace, and V. indica also forage rotting food. The PERs to 52 tested compounds were in good agreement with the food habits of these butterflies. The six species showed high PERs to several aromatic compounds widely found in flower scents. Fermentation products such as ethanol and acetic acid elicited intermediate to high responses from the three species feeding on rotting foods. The other three species showed low PERs to these compounds, and acetic acid strongly inhibited the PER-eliciting activities of the other compounds. These results demonstrate that the olfactory preference for fermentation products is a characteristic of the rotting-food feeders. The spectrum of EAG responses to 37 tested compounds was relatively similar among the six species. Particular acyclic aliphatic compounds, including green leaf volatiles, elicited high EAG responses from all the species. Despite a high PER-eliciting activity, most aromatic compounds induced intermediate EAG responses; however, ethanol and acetic acid evoked very weak responses. The results suggest that in food selection, adult butterflies use a specific olfactory system to perceive these major food-derived volatiles.     

Behavioural and electrophysiological responses of the olive fruit fly, Bactrocera oleae (Rossi) (Diptera: Tephritidae), to male- and female-borne sex attractants

In the olive fruit fly, Bactrocera oleae, females attract males by producing 1,7-dioxaspiro[5.5]undecane (olean), the main component of the sex pheromone secreted by rectal glands. It has been recently demonstrated that males are able to produce (Z)-9-tricosene (muscalure) in rectal glands, a compound that selectively attracts females. In this study, a male grooming reaction that may transfer the male-borne compounds from rectal to urotergal glands was observed, suggesting that urotergal glands could be involved in B. oleae sexual communication. GC/MS, EAG, GC/EAD analyses and behavioural assays were carried out to compare the role of male rectal and urotergal glands during courtship. In both male glands, olean and muscalure amounts were age dependent. Extracts of rectal glands contained higher amounts of olean and/or muscalure than urotergal ones. Extracts of rectal and urotergal glands of males and females elicited EAG responses in both sexes. GC/EAD showed that female EAG response to male rectal extracts was mainly due to olean and muscalure. Synthetic compounds evoked EAG dose-dependent responses in both sexes, and the EAG response to muscalure was higher as compared to olean. Rectal and urotergal glands from old males were able to attract females, while urotergal glands from young males attracted only males. Overall, our results add knowledge to the mating system of B. oleae, giving first evidences on the electrophysiological activity of muscalure towards both sexes, as well as on the involvement of male urotergal glands in the chemical sexual communication of this pest.     

Electrophysiological responses and field attraction of the grey corn weevil, Tanymecus (Episomecus) dilaticollis Gyllenhal (Coleoptera: Curculionidae) to synthetic plant volatiles

The grey corn weevil, Tanymecus (Episomecus) dilaticollis, is an economically important polyphagous pest in Eastern and Central Europe. The present research is the first published electrophysiological study of olfactory sensitivity of adult T. dilaticollis to plant volatiles. The electrophysiological responses of male and female antennae were recorded to 20 synthetic volatile organic compounds (VOCs), previously identified to be released from the preferred host plants, maize and sunflower. Antennae of both sexes responded to a wide range of volatiles from different chemical classes: fatty acid derivatives, aromatic compounds and terpenoids. Two green leaf volatiles, (E)-2-hexen-1-ol and (Z)-3-hexen-1-ol, and one terpenoid, (±)-linalool, elicited the strongest electrophysiological responses by male and female antennae. Relatively high electrophysiological activity by T. dilaticollis antennae was also recorded to benzaldehyde, eugenol and caryophyllene. In the field, (E)-2-hexen-1-ol and (Z)-3-hexen-1-ol, when presented individually, caught significantly more male and female T. dilaticollis than unbaited controls. These compounds could be developed into an attractive trap for the detection and monitoring of T. dilaticollis.     

Attraction of two larval parasitoids with varying degree of host specificity to single components and a binary mixture of host-related plant volatiles

As an indirect defense to herbivore attack, plants release many types of volatile organic compounds (VOCs), which guide parasitoids to their herbivore hosts. In the present study, VOCs were categorized as those released passively from undamaged plants and herbivore-induced plant volatiles (HIPVs). HIPVs were further categorized into: (1) volatiles released by fresh damage plants, and (2) volatiles released by old damage plants. We used as models, two parasitoids with different degree of host specificity, Microplitis croceipes (specialist) and Cotesia marginiventris (generalist), to address the evolutionary and mechanistic question of whether specialist and generalist parasitoids differ in their use of VOCs for host location. Both species are solitary larval endoparasitoids in the same family (Hymenoptera: Braconidae) and are important parasitoids of caterpillar pests of cotton. Based on the results of previous studies, α-pinene, (Z)-3-hexenol, and (Z)-3-hexenyl acetate were selected as representatives of the different types of VOCs in cotton. The attraction of both parasitoid species to synthetic components and a binary mixture of the above VOCs was tested in four-choice olfactometer bioassays. Female M. croceipes showed the greatest attraction to the HIPVs while female C. marginiventris could not discriminate among the three VOCs. Conspecific males showed similar responses with a few exceptions. When presented with the choices; α-pinene, (Z)-3-hexenol and a binary mixture (50:50v/v) of the two compounds, the specialist showed the greatest attraction to the mixture. However, the mixture did not elicit such an additive effect on the attraction of the generalist. Overall response latency (time taken to choose VOCs) indicated species and sexual (in the specialist) differences. Using a simple model, this study provides a fundamental insight into odor preferences and discriminatory ability of the test parasitoids. The ecological significance and practical implications of these results are discussed.     

The olfactory response of the reindeer nose bot fly,Cephenemyia trompe (Oestridae), to components from interdigital pheromone gland and urine from the host reindeer,Rangifer tarandus

Summary The linked gas chromatographical/electroan-tennogram (GC/EAG) technique revealed that the parasitic reindeer nose bot fly is able to specifically sense components produced by the interdigital pheromone gland of reindeer. The head-space extraction technique, with Porapak Q as the collecting polymer, was used to trap pheromone gland and urine components used to assess fly responses. One component from reindeer urine also was a potent stimulus for the sensory neurons of the fly. These components can be important chemical signals to the flies for long distance orientation towards host animals. This is the first report on EAG in Oestridae.     

Sensory cues associated with host detection in a marine parasitic isopod

Parasite population dynamics and the evolution of life history characteristics are strongly associated with the process of host infection. Parasites with free-living life stages have a narrow window to infect a host and have evolved a number of mechanisms to detect and locate a host. Cymothoa excisa is a parasitic isopod that is commonly found on the Atlantic croaker, Micropogonias undulatus, in the Gulf of Mexico. Here, we determined the infection window that constrains host-searching behavior in C. excisa and tested the behavioral response of the free-swimming larvae (mancae) to visual and chemical cues. Mancae were found to have an infection window of 7 days. Mancae were responsive to both visual and chemical cues. Our findings are the first to show that cymothoid isopods use visual and chemical cues to locate a host and that individuals display a host-locating strategy that maximizes host encounter rate, while reducing energy expenditure.     

Pathogen-induced hatching and population-specific life-history response to waterborne cues in brown trout (Salmo trutta)

Hatching is an important niche shift, and embryos in a wide range of taxa can either accelerate or delay this life-history switch in order to avoid stage-specific risks. Such behavior can occur in response to stress itself and to chemical cues that allow anticipation of stress. We studied the genetic organization of this phenotypic plasticity and tested whether there are differences among populations and across environments in order to learn more about the evolutionary potential of stress-induced hatching. As a study species, we chose the brown trout (Salmo trutta; Salmonidae). Gametes were collected from five natural populations (within one river network) and used for full-factorial in vitro fertilizations. The resulting embryos were either directly infected with Pseudomonas fluorescens or were exposed to waterborne cues from P. fluorescens-infected conspecifics. We found that direct inoculation with P. fluorescens increased embryonic mortality and induced hatching in all host populations. Exposure to waterborne cues revealed population-specific responses. We found significant additive genetic variation for hatching time, and genetic variation in trait plasticity. In conclusion, hatching is induced in response to infection and can be affected by waterborne cues of infection, but populations and families differ in their reaction to the latter.     

Ubiquitous eight-carbon volatiles of fungi are infochemicals for a specialist fungivore

An enormous variety of volatile substances are released in distinctive blends by fungal substrates that should be recognisable for fungivores. Certain compounds dominate in most of the fungal species. Fungal oxylipins as the eight-carbon volatiles are the most prominent. This raises the question whether such are specific enough to qualify as appropriate host cues for a fungivore. We could demonstrate differentiated responses of the fungivorous beetle Bolitophagus reticulatus to eight-carbon volatiles: Nine eight-carbon volatiles were identified with GC–MS from its host fungus Fomes fomentarius. 1-Octen-3-ol, 3-octanone and 3-octanol induced contrasting behaviour of beetles in olfactometer bioassays. Electroantennographic experiments investigating the beetle olfactory sense revealed distinguishable antennal responses. Moreover, their individual release from F. fomentarius fruiting bodies changes not only considerably, but also independently over successive stages of beetle colonisation. Concentrations of attractive and repellent eight-carbon volatiles correlate to frequency of beetles in the field and further substantiate their relevance as host cues. Our results show that a specialist fungivore is able to differentiate the most common eight-carbon volatiles of fungi to assess host quality. Key roles and marked similarities of fungal to plant oxylipins suggest a comparable importance of eight-carbon volatiles to fungivores as green leaf volatiles have to herbivores.     

Monitoring of Bt-Maize pollen exposure in the vicinity of the nature reserve Ruhlsdorfer Bruch in northeast Germany 2007 to 2008

Background, aim, and scope Basically, technological innovations are associated with benefits and risks. This is also true for the introduction of genetically modified organisms (GMO) into agriculture. In Germany, precautionary regulations currently demand isolation distances (i.?e. buffer zones) for the cultivation of genetically modified maize (Bt-maize) in the vicinity of conventional (150?m) and organic maize fields (300?m). The Bt-toxin may harm non-target organisms (NTO) such as Lepidoptera. Despite this, corresponding regulations for the protection of nature reserves are lacking to date. Conventional and Bt-maize have been grown in the vicinity of the Flora-Fauna-Habitat (FFH) Ruhlsdorfer Bruch in Brandenburg, Germany. The aim of this study was to investigate whether exposure of maize pollen from surrounding Bt-maize fields to NTOs in the nature protection area could be excluded or not. Two types of exposure were investigated: Firstly, whether maize pollen was dispersed by wind into the nature reserve area exposing resident NTOs. Secondly, foraging NTOs from the nature reserve are exposed by roaming the surrounding fields and collecting maize pollen. In order to fulfil the precautionary principle defined by law, the study should help to determine appropriate isolation distances for the cultivation of Bt-maize with regard to sustainable protection of NTOs in the FFH Ruhlsdorfer Bruch. In 2007, the local authorities issued an isolation distance between Bt-maize fields and nature reserves of 100?m and in 2008, this became 250?m in the northern and 500?m in the westerly direction, respectively. Materials and methods Standardised methods for biological and technical pollen sampling issued by the Association of German Engineers (VDI 4330 Part 3, 2007 and VDI 4330 Part 4, 2006) were applied providing a quality controlled and methodologically harmonised database which does not only serve the needs to be fulfilled by the present case-specific monitoring study but can also be used as a reference database for further investigations. Maize pollen exposure was measured within the FFH Ruhlsdorfer Bruch and its immediate vicinity in July and August 2007 and 2008. In 2007, the sampling was performed at three sites using 12 technical samplers (Sigma-2/PMF) placed at five measuring points at distances from 5?m to 120?m from the maize field edges. Additionally, for biological pollen sampling six bee colonies were situated at these three sites (two colonies at each site). The technical sampler Sigma-2/PMF enables point sampling which is primarily influenced by wind and topography providing information on effective maize pollen input (flow and deposition) at the measuring sites. Honey-bees roaming in the surrounding area with typical foraging distances of several kilometres may act as planar collectors. They may serve as indicators for the exposure to pollen-collecting NTOs. Furthermore, biological sampling is more selective due to the organism’s preferences, whereas the technical sampling is neutral. Hence, both the technical and the biological sampling complement each other in their scope of application. The pollen samples of both matrices were analysed microscopically and the maize pollen loads were quantified. Pollen-DNA was analysed by means of the quantitative PCR-method (qPCR) identifying conventional and Bt-maize pollen by two independent laboratories. In 2008, the monitoring was repeated with additional sites. Eighteen technical samplers were exposed at five sites with eight measuring points at distances from 5?m to 250?m from the maize field edges. Two honey-bee colonies for biological sampling were placed at one site for control purposes. PCR-analyses were performed to measure the amounts of Bt-maize pollen in the samples. Results The results of pollen monitoring at the Ruhlsdorfer Bruch revealed maize pollen exposure for all monitoring sites in both surveys. In 2007, up to 1.75 million maize pollen/m² were deposited at sites closest to the maize field. At 120?m from field edge in the middle of the FFH-reserve, 99,000 maize pollen/m² were detected. In 2008, similar results were found, at distances up to 250?m from the field edges deposition of 164,000 maize pollen/m² was detected. Data on maize pollen deposition show a clear distance relationship and are in accordance with results of further comprehensive surveys based on the same methodology (Hofmann 2007). The results of the microscopic analysis of the pollen pellets demonstrated that bees collected maize pollen at all sites, in 2007 and 2008. Although maize pollen is not the main food source (2007:0.1–0.3?%; 2008:2–3?%) the collection efficiency of the bee colonies resulted in high amounts of sampled maize pollen with 4 to 11 million per site in 2007 and up to 467 million in 2008. Molecular-biological analysis of maize pollen DNA by qPCR demonstrated that transgenic Bt-MON810 DNA was present in all technical and biological samples, corroborated by two independent laboratories. In 2007, the GMO-content in the samples ranged up to 44?% in the bioaerosols and 49?% in the pollen pellets. In 2008, GMO-proportions of up to 18?% were detected. Discussion The results of this study provide evidence that NTOs in the Ruhlsdorfer Bruch were exposed to Bt-maize pollen under the cultivation conditions in 2007 with a buffer zone of 100?m. The GMO-content reached up to 48?%. The results of the monitoring in 2008 confirmed these findings. Even though the exposure could be reduced by increasing the isolation distances to 250?m and 500?m respectively, the results still show percentages of up to 18?% Bt-MON810 in the pollen samples. The results on maize pollen deposition at the Ruhlsdorfer Bruch in 2007 and 2008 correspond to the results of an investigation which was conducted over several years applying the same standardised method, but covering a wider range of distances. The correlation between maize pollen deposition (n/m²) and distance to the source field (m) fitted best to a power function of the type y = 1.2086 · 10⁶ · x ^–0.548. Despite the same trend, the pollen deposition in the Ruhlsdorfer Bruch revealed above-average findings. Also the analysis of the pollen pellets collected by the bees showed an exposure in 2007 with values for the GMO-content of up to 49?%. For both methods, the exposure decreased in 2008 due to the greater buffer zones up to 500?m. Whereas the GMO-content for the biological sampling were reduced to values below 10?%, the values for the technical sampling were still higher indicating that greater buffer zones would be necessary for safety reasons under the precautionary principle. Conclusions The results of this investigation proved that maize pollen were dispersed by wind to distances farther than 250?m from field edge leading to maize pollen exposure in the centre of the nature reserve. The results also demonstrated that foraging NTOs living in the nature reserve were exposed to maize pollen from surrounding fields. Considering the cultivation of Bt-maize MON810, the assumption of the Environmental Risk Assessment (ERA) that there will be no relevant exposure beyond the Bt-maize fields, cannot be confirmed. Considering the results of this and related studies and with respect to the precautionary principle, one can state that buffer zones between Bt-maize fields and protected areas are an effective measure to minimise the exposure of Bt-maize pollen to NTOs and, thus, to prevent from adverse effects. Recommendations and perspectives Because of still insufficient ecotoxicological data for the risk assessment of Bt-maize MON810 considering butterflies and other protected NTOs, protection standards assuring the precautionary principle have to be implemented to avoid Bt-maize pollen exposure to NTOs. This applies for the case Ruhlsdorfer Bruch and for nature reserve areas in general. In order to exclude risks to protected NTOs occurring in nature reserves, sufficient buffer zones for Bt-maize cultivation should be considered. The statistical analysis revealed that distances of more than 500?m are necessary to decisively reduce exposure to foraging insects. In fact, distances of more than 1,000?m are necessary to prevent maize pollen deposition from values above 100,000 pollen/m² with a certainty of 90?%. An adequate risk assessment can only be attained if based on field measurements accounting for the high variation of pollen deposition due to local environmental site conditions and field management. The monitoring should be based on standardised methods. It should include locations with the highest expected deposition rates, the boundaries of the protected areas and sites of interest within those boundaries, e.?g., specific habitats of sensitive species.     

Antipredator responses to invasive lionfish, Pterois volitans: interspecific differences in cue utilization by two coral reef gobies

Understanding prey response to predators and their utilization of sensory cues to assess local predation risk is crucial in determining how predator avoidance strategies affect population demographics. This study examined the antipredator behaviors of two ecologically similar species of Caribbean coral reef fish, Coryphopterus glaucofraenum and Gnatholepis thompsoni, and characterized their responses to different reef predators. In laboratory assays, the two species of gobies were exposed to predator visual cues (native Nassau grouper predator vs. invasive lionfish predator), damage-released chemical cues from gobies, and combinations of these, along with appropriate controls. Behavioral responses indicate that the two prey species differ in their utilization of visual and chemical cues. Visual cues from predators were decisive for both species’ responses, demonstrating their relative importance in the sensory hierarchy, whereas damage-released cues were a source of information only for C. glaucofraenum. Both prey species could distinguish between native and invasive predators and subsequently altered their antipredator responses.     

Sex pheromone components of pyralid moths Terastia subjectalis and Agathodes ostentalis feeding on the coral tree, Erithrina variegata: Two sympatric species share common components in different ratios

Summary. Two common components, identified as (E)-11-hexadecenal (E11-16:Ald) and (E,E)-10,12-hexadecadienal (E10E12-16:Ald), were found in the extract of virgin females of two sympatric pyralid moths, Terastia subjectalis Lederer, and Agathodes ostentalis (Geyer). The amount of E11-16:Ald and E10E12-16:Ald was 0.12 and 2.8 ng/female for T. subjectalis and 2.0 and 1.8 ng/female for A. ostentalis, respectively. Hexadecanal (16:Ald) was also found at 0.7 ng/female in the extract of A. ostentalis, but EAG activity was unclear. In the field, T. subjectalis and A. ostentalis males were captured with species specific blends in the proportions of 5:95 and 50:50 blends. Blends attractive to one species did not attract the opposite one. Rubber septa loaded with 1 mg each of 5:95 and 50:50 blends of E11-16:Ald and E10E12-16:Ald were as attractive as two virgin females of T. subjectalis and A. ostentalis , respectively. This demonstrated that different ratios of shared pheromone components could provide species specific cues for critical mate location in the two sympatric pyralid moths. It is suggested that directional selection probably occurs in the pheromone blends in opposite directions in both species.     

Parasitism at the Landscape Scale: Cowbirds Prefer Forests

Landscape-scale examination of parasitism patterns of Brown-headed Cowbirds ( Molothrus ater ) revealed heterogeneous parasitism rates across the mosaic of a forest and associated old-field communities. In a two year study in Dutchess County, New York, we found a significantly higher parasitism rate in the forest-interior community ( n = 301 nests; 17 species) than on the species in the adjacent and nearby old field and edge ( n = 328 nests; 15 species; 32.3% versus 6.5%; p < 0.0001). Cowbirds invaded a mature 1300-ha forest stand even when their traditional host species were available in adjacent old-field and edge habitats. The forest and old-field study areas were located in a 38,000-ha township with 55% forest cover and contained numerous agriculture, dairy, and horse farms that provided favorable habitat for cowbirds. Within-forest examination of parasitism patterns revealed four aspects of cowbird parasitism that contrasted with patterns described in other regions: (1) parasitism was concentrated significantly more often on ground- and low-nesting (nests ≤ 1 m) forest species than on medium- and high-nesting species (nests> 1 m; 35.01% versus 29.93%; p = 0.0393); (2) parasitism was not significantly greater on Neotropical migrant species than on short-distance migrants and residents; (3) the parasitism rate was not higher in nests close to edges; and (4) the parasitism level was low on certain forest species (such as Wood Thrush) that have experienced high parasitism levels in the Midwest. From a management perspective these data suggest that cowbirds exhibit regional differences in host and habitat use; the target host community of a particular cowbird population is unpredictable at the landscape scale; and a landscape scale should be used in designing cowbird studies to accurately assess local population dynamics.     

Effects of phosphorus sources on volatile organic compound emissions from <Emphasis Type="Italic">Microcystis flos</Emphasis>-<Emphasis Type="Italic">aquae</Emphasis> and their toxic effects on <Emphasis Type="Italic">Chlamydomonas reinhardtii</Emphasis>

There is diverse phosphorus (P) in eutrophicated waters, but it is considered as a crucial nutrient for cyanobacteria growth due to its easy precipitation as insoluble salts. To uncover the effects of complex P nutrients on the emission of volatile organic compounds (VOCs) from cyanobacteria and their toxic effects on other algae, the VOCs from Microcystis flos-aquae supplied with different types and amount of P nutrients were analyzed, and the effects of VOCs and their two main compounds on Chlamydomonas reinhardtii growth were investigated. When M. flos-aquae cells were supplied with K₂HPO₄, sodium pyrophosphate and sodium hexametaphosphate as the sole P source, 27, 23 and 29 compounds were found, respectively, including furans, sulfocompounds, terpenoids, benzenes, aldehydes, hydrocarbons and esters. With K₂HPO₄ as the sole P source, the VOC emission increased with reducing P amount, and the maximum emission was found under Non-P condition. In the treatments of M. flos-aquae VOCs under Non-P condition and two main terpenoids (eucalyptol and limonene) in the VOCs, remarkable decreases were found in C. reinhardtii cell growth, photosynthetic pigment content and photosynthetic abilities. Therefore, we deduce that multiple P nutrients in eutrophicated waters induce different VOC emissions from cyanobacteria, and P amount reduction caused by natural precipitation and algal massive growth results in more VOC emissions. These VOCs play toxic roles in cyanobacteria becoming dominant species, and eucalyptol and limonene are two toxic agents.