Anopheles arabiensis oviposition site selection in response to habitat persistence and associated physicochemical parameters, bacteria and volatile profiles.

A better understanding of the oviposition behaviour of malaria vectors might facilitate the development of new vector control tools. However, the factors that guide the aquatic habitat selection of gravid females are poorly understood. The present study explored the relative attractiveness of similar artificial ponds (0.8 m2 ) aged at varying lengths prior to opening in such a way that wild Anopheles arabiensis could choose between ponds that were freshly set up, or were aged 4 or 17 days old, to lay eggs. Physicochemical parameters, bacterial profile and volatile organic compounds emitted from ponds were investigated over three experimental rounds. Fresh ponds contained on average twice as many An. arabiensis instar larvae (mean 50, 95% confidence interval (CI) = 29-85) as the ponds that had aged 4 days (mean = 24, 95% CI = 14-42) and 17 days (mean = 20, 95% CI: 12-34). Fresh ponds were associated with a significantly higher turbidity combined with higher water temperature, higher nitrite levels and a lower pH and chlorophyll level than the older ponds. Round by round analyses suggested that bacteria communities differed between age groups and also that 4-heptanone, 2-ethylhexanal and an isomer of octenal were exclusively detected from the fresh ponds. These characteristics may be useful with respect to developing attract and kill strategies for malaria vector control.

Dynamics of putative sex pheromone components during heat periods in estrus-induced cows.

Determination of the optimal insemination time in dairy cows is vital for fertilization success and is a challenging task due to silent or weak signs of estrus shown by some cows. This can be overcome by combining several estrus detection methods, leading to higher detection rates. However, an efficient, noninvasive method for detecting estrus in cows is still needed. Chemical cues released by the cow during estrus have been proposed to have pheromonal properties and signal readiness to mate to the bull. Such cues could be used in an industrial setting to detect cows in estrus. However, no conclusive published data show temporal changes in putative sex pheromone levels during estrus. The goal of this study was to determine the temporal pattern of putative sex pheromone components during estrus and to assess the reproducibility of changes in pheromone concentration with respect to ovulation time. Two injections of the hormone PGF2α were administered over a 2-wk interval to induce and synchronize the estrous cycles of 6 Holstein cows. The precise time of ovulation was determined by means of an ultrasound technique, and estrus was determined by visual observation. Using solid-phase microextraction gas chromatography-mass spectrometry techniques, we showed that acetic and propionic acids, which have been proposed to be putative sex pheromone components in cows, were present in the headspaces of all estrous and diestrous fecal samples, whereas 1-iodoundecane was not detected by solid-phase microextraction or by solvent extraction with diethyl ether. Low levels of acids were observed until 1 d before ovulation, at which point their concentrations increased, peaking around 0.5 d before ovulation. The application of labeled synthetic standards revealed that during the peak of release, 36 ± 8 ng (average ± SD) of acetic acid and 10 ± 3 ng of propionic acid were present in 0.5-g samples of estrous-phase fecal matter compared with 19 ± 5 and 2.3 ± 1 ng of acetic and propionic acids, respectively, in the control diestrous samples. After the peak, the amounts of the compounds decreased sharply to match those of the control samples and afterward returned to the baseline readings. This decrease in the amounts of putative pheromone components was registered about 12 h before ovulation, indicating that acetic and propionic acids could be used as biomarkers for the electronic detection of ovulation.

Activity studies of sesquiterpene oxides and sulfides from the plant Hyptis suaveolens (Lamiaceae) and its repellency on Ixodes ricinus (Acari: Ixodidae).

Hyptis suaveolens (Lamiaceae), a plant traditionally used as a mosquito repellent, has been investigated for repellent properties against nymphs of the tick Ixodes ricinus. Essential oils and volatile compounds of fresh and dried leaves, from plants originating from Laos and Guinea-Bissau, were identified by GC-MS and tested in a tick repellency bioassay. All the essential oils were strongly repellent against the ticks, even though the main volatile constituents differed in their proportions of potentially tick repellent chemicals. (+)/(-)-sabinene were present in high amounts in all preparations, and dominated the emission from dry and fresh leaves together with 1,8-cineol and α-phellandrene. 1,8-Cineol and sabinene were major compounds in the essential oils from H. suaveolens from Laos. Main compounds in H. suaveolens from Guinea-Bissau were (-)-sabinene, limonene and terpinolene. Among the sesquiterpene hydrocarbons identified, α-humulene exhibited strong tick repellency (96.8 %). Structure activity studies of oxidation or sulfidation products of germacrene D, α-humulene and ß-caryophyllene, showed increased tick repellent activity: of mint sulfide (59.4 %), humulene-6,7-oxide (94.5 %) and caryophyllene-6,7-oxide (96.9 %). The substitution of oxygen with sulfur slightly lowered the repellency. The effects of the constituents in the oils can then be regarded as a trade off between the subsequently lower volatility of the sesquiterpene derivatives compared to the monoterpenes and may thus increase their potential usefulness as tick repellents.

Primary and secondary organics in the tropical Amazonian rainforest aerosols: chiral analysis of 2-methyltetraols.

This work presents the application of a new method to facilitate the distinction between biologically produced (primary) and atmospherically produced (secondary) organic compounds in ambient aerosols based on their chirality. The compounds chosen for this analysis were the stereomers of 2-methyltetraols, (2R,3S)- and (2S,3R)-methylerythritol, (l- and d-form, respectively), and (2S,3S)- and (2R,3R)-methylthreitol (l- and d-form), shown previously to display some enantiomeric excesses in atmospheric aerosols, thus to have at least a partial biological origin. In this work PM10 aerosol fractions were collected in a remote tropical rainforest environment near Manaus, Brazil, between June 2008 and June 2009 and analysed. Both 2-methylerythritol and 2-methylthreitol displayed a net excess of one enantiomer (either the l- or the d-form) in 60 to 72% of these samples. These net enantiomeric excesses corresponded to compounds entirely biological but accounted for only about 5% of the total 2-methyltetrol mass in all the samples. Further analysis showed that, in addition, a large mass of the racemic fractions (equal mixtures of d- and l-forms) was also biological. Estimating the contribution of secondary reactions from the isomeric ratios measured in the samples (=ratios 2-methylthreitol over 2-methylerythritol), the mass fraction of secondary methyltetrols in these samples was estimated to a maximum of 31% and their primary fraction to a minimum of 69%. Such large primary fractions could have been expected in PM10 aerosols, largely influenced by biological emissions, and would now need to be investigated in finer aerosols. This work demonstrates the effectiveness of chiral and isomeric analyses as the first direct tool to assess the primary and secondary fractions of organic aerosols.

Tick repellent properties of three Libyan plants.

Specimens of Salvadora persica Wallich (Salvadoraceae), Pistacia atlantica Desfontaines (Anacardiaceae), and Juniperus phoenicea L. (Cupressaceae) were collected at The Green Mountain in northeastern Libya. The tick-repellent effects of the essential oils from the plants were evaluated using host-seeking nymphs of Ixodes ricinus L. (Acari: Ixodidae) in the laboratory. Significant tick repellent effects were observed for the oils of all three species, but the duration of action was short. The oils were obtained by steam distillation. Volatile compounds of the essential oils were collected by solid phase microextraction and the constituents were identified by gas chromatographymass spectrometry.

Oviposition responses of Anopheles gambiae s.s. (Diptera: Culicidae) and identification of volatiles from bacteria-containing solutions.

In this study, a dual-choice oviposition bioassay was used to screen responses of gravid An. gambiae toward 17 bacterial species, previously isolated from Anopheles gambiae s.l. (Diptera: Culicidae) midguts or oviposition sites. The 10 isolates from oviposition sites have been identified by phylogenetic analyses of their 16S rRNA genes. Eight of the 10 isolates were gram-positive, out of which six belonged to the Bacilli class. Solid phase microextraction and gas chromatography coupled to mass spectrometry (GC-MS) were used to identify the volatiles emitted from the bacterial isolates. Aromatic and aliphatic alcohols, aliphatic ketones, alkylpyrazines, dimethyl oligosulfides, and indole were among the chemical compounds identified from the headspace above bacteria-containing saline. The mosquitoes laid significantly more eggs in six of the bacteria-containing solutions compared with the sterile solution. These six bacteria did not emit any compounds in common that could explain the positive oviposition response. Instead, the bacteria were grouped according to principal component analysis (PCA) based on the relative amounts of volatiles emitted. The PCA-plots facilitated the identification of 13 putative oviposition attractants for An. gambiae mosquitoes.

Transstadial and horizontal transfer of bacteria within a colony of Anopheles gambiae (Diptera: Culicidae) and oviposition response to bacteria-containing water.

In a paratransgenic approach, genetically modified bacteria are utilized to kill the parasite in the vector gut. A critical component for paratransgenics against malaria is how transgenic bacteria can be introduced and then kept in a mosquito population. Here, we investigated transstadial and horizontal transfer of bacteria within an Anopheles gambiae mosquito colony with the focus on spiked breeding sites as a possible means of introducing bacteria to mosquitoes. A Pantoea stewartii strain, previously isolated from An. gambiae, marked with a green fluorescent protein (GFP), was introduced to mosquitoes in different life stages. The following life stages or older mosquitoes in the case of adults were screened for bacteria in their guts. In addition to P. stewartii other bacteria were isolated from the guts: these were identified by 16S rRNA sequence analysis and temporal temperature gradient gel electrophoresis (TTGE). Bacteria were transferred from larvae to pupae but not from pupae to adults. The mosquitoes were able to take up bacteria from the water they emerged from and transfer the same bacteria to the water they laid eggs in. Elizabethkingia meningoseptica was more often isolated from adult mosquitoes than P. stewartii. A bioassay was used to examine An. gambiae oviposition responses towards bacteria-containing solutions. The volatiles emitted from the solutions were sampled by headspace-solid phase microextraction (SPME) and identified by gas chromatography and mass spectrometry (GC-MS) analysis. P. stewartii but not E. meningoseptica mediated a positive oviposition response. The volatiles emitted by P. stewartii include indole and 3-methyl-1-butanol, which previously have been shown to affect An. gambiae mosquito behaviour. E. meningoseptica emitted indole but not 3-methyl-1-butanol, when suspended in saline. Taken together, this indicates that it may be possible to create attractive breeding sites for distribution of genetically modified bacteria in the field in a paratransgenic approach against malaria. Further research is needed to determine if the bacteria are also transferred in the same way in nature.

Plant volatiles activating specific olfactory receptor neurons of the cabbage moth Mamestra brassicae L. (Lepidoptera, Noctuidae).

Herbivore insects are suitable model organisms for studying how plant odor information is encoded in olfactory receptor neurons (RNs). By the use of gas chromatography linked to electrophysiological recordings from single RNs, screening for sensitivity to naturally produced plant odorants is possible in order to determine the molecular receptive ranges of the neurons. Using this method, we have in this study of the cabbage moth, Mamestra brassicae, classified 21 types of olfactory RNs according to their responses to odorants present in the host plants of Brassicae, in the related species of Arabidopsis, as well as in essential oils of nonhost plants like ylang-ylang. Most of the RNs were tuned to one or a few structurally similar compounds, showing minimal overlap of their molecular receptive ranges. Whereas some RNs displayed a novel tuning, others were tuned to the same compounds as neurons in other insect species. We also found colocation in the same sensillum of 3 RN types with the same response characteristics and tuning as 3 colocated types described in heliothine moths living on different host plants. The presence of similar RN types across different insect species implies conservation or reappearance of the RN types, independent of the evolution of host plant ranges.

Structure-activity relationships of phenylpropanoids as antifeedants for the pine weevil Hylobius abietis.

Ethyl cinnamate has been isolated from the bark of Pinus contorta in the search for antifeedants for the pine weevil, Hylobius abietis. Based on this lead compound, a number of structurally related compounds were synthesized and tested. The usability of the Topliss scheme, a flow diagram previously used in numerous structure-activity relationship (SAR) studies, was evaluated in an attempt to find the most potent antifeedants. The scheme was initially followed stepwise; subsequently, all compounds found in the scheme were compared. In total, 51 phenylpropanoids were tested and analyzed for SARs by using arguments from the field of medicinal chemistry (rational drug design). Individual Hansch parameters based on hydrophobicity, steric, and electronic properties were examined. The effects of position and numbers of substituents on the aromatic ring, the effects of conjugation in the molecules, and the effects of the properties of the parent alcohol part of the esters were also evaluated. It proved difficult to find strong SARs derived from single physicochemical descriptors, but our study led to numerous new, potent, phenylpropanoid antifeedants for the pine weevil. Among the most potent were methyl 3-phenylpropanoates monosubstituted with chloro, fluoro, or methyl groups and the 3,4-dichlorinated methyl 3-phenylpropanoate.

Methyl salicylate, identified as primary odorant of a specific receptor neuron type, inhibits oviposition by the moth Mamestra brassicae L. (Lepidoptera, noctuidae).

The cabbage moth, Mamestra brassicae L. (Lepidoptera, Noctuidae), is a polyphagous species that is often choosing plants of Brassica as hosts for oviposition. In the search for biologically relevant odorants used by these moths, gas chromatography linked to electrophysiological recordings from single receptor neurons (RNs) has been employed, resulting in classification of distinct types of neurons. This study presents specific olfactory RNs responding to methyl salicylate (MeS) as primary odorant and showing a weak response to methyl benzoate, the 2 aromatic compounds occurring together in several plant species. In 2 cases, the neuron was colocated with another RN type responding to 6 green leaf volatiles: 1-hexanol, (3Z)-hexen-1-ol, (2E)-hexen-1-ol, (3Z)-hexenyl acetate, (2Z)-hexen-1-ol, and an unidentified compound. Whereas the specific RNs detected the minor amounts of MeS in some plants, the compound was not found by gas chromatography linked to mass spectrometry in intact plants, but it was found after herbivore attack. The behavioral effect of MeS was studied in outdoor test arenas with Brassica napus and artificial plants. These experiments indicated that mated M. brassicae females avoid plants with dispensers emitting MeS. As it is induced by caterpillar feeding, this compound may mediate a message to mated M. brassicae females that the plant is already occupied.

Discrimination between enantiomers of linalool by olfactory receptor neurons in the cabbage moth Mamestra brassicae (L.).

Plants emit complex blends of volatiles, including chiral compounds that might be detected by vertebrates and invertebrates. Insects are ideal model organisms for studying the underlying receptor neuron mechanisms involved in olfactory discrimination of enantiomers. In the present study, we have employed two-column gas chromatography linked to recordings from single olfactory receptor neurons of Mamestra brassicae, in which separation of volatiles in a polar and a chiral column was performed. We here present the response properties of olfactory receptor neurons tuned to linalool. The narrow tuning of these receptor neurons was demonstrated by their strong responses to (R)-(-)-linalool, the weaker responses to the (+)-enantiomer as well as a few structurally related compounds, and no responses to the other numerous plant released volatiles. The enantioselectivity was verified by parallel dose-response curves, that of (R)-(-)-linalool shifted 1 log unit to the left of the (S)-(+)-linalool curve. A complete overlap of the temporal response pattern was found when comparing the responses of the same strength. Analysis of the spike amplitude and waveform indicated that the responses to the two enantiomers originated from the same neuron.

Evaluation of extracts and oils of tick-repellent plants from Sweden.

Abstract. Leaves of Myrica gale Linnaeus (Myricaceae), Rhododendron tomentosum (Stokes) H. Harmaja (formerly Ledum palustre Linnaeus: Ericaceae) and Artemisia absinthium Linnaeus (Asteraceae) were extracted with organic solvents of different polarities and the essential oils of leaves were obtained by steam distillation. The extracts or oils were tested in the laboratory for repellency against host-seeking nymphs of Ixodes ricinus Linnaeus (Acari: Ixodidae). Rhododendron tomentosum oil, 10%, diluted in acetone, exhibited 95% repellency; R. tomentosum and A. absinthium extracts in ethyl acetate, > 70% repellency; A. absinthium extract in hexane, approximately 62% repellency; and M. gale oil, 10%, approximately 50% repellency on I. ricinus nymphs. Compounds in the leaf extracts or in the oils were collected by solid phase microextraction (SPME) and identified by gas chromatography-mass spectrometry (GC-MS) and/or MS. Characteristic volatiles detected from oil or extract of M. gale were the monoterpenes 1,8-cineole, alpha-terpineol, 4-terpineol and thujenol; and of R. tomentosum myrcene and palustrol. Characteristic volatiles from leaf extracts of A. absinthium were sabinene, oxygenated monoterpenes, e.g. thujenol and linalool, and geranyl acetate. Each plant species synthesized numerous volatiles known to exhibit acaricidal, insecticidal, 'pesticidal' and/or arthropod repellent properties. These plants may be useful sources of chemicals for the control of arthropods of medical, veterinary or agricultural importance.

Olfactory receptor neurons in two Heliothine moth species responding selectively to aliphatic green leaf volatiles, aromatic compounds, monoterpenes and sesquiterpenes of plant origin.

Moths of the subfamily Heliothinae are suitable models for comparative studies of plant odour information encoded by the olfactory system. Here we identify and functionally classify types of olfactory receptor neurons by means of electrophysiological recordings from single receptor neurons linked to gas chromatography and to mass spectrometry. The molecular receptive ranges of 14 types in the two polyphagous species Heliothis virescens and Helicoverpa armigera are presented. The receptor neurons are characterized by a narrow tuning, showing the best response to one primary odorant and weak responses to a few chemically related compounds. The most frequently occurring of the 14 types constituted the receptor neurons tuned to (+)-linalool, the enantioselectivity of which was shown by testing two samples with opposite enantiomeric ratios. These neurons, also responding to dihydrolinalool, were found to be functionally similar in the two related species. The primary odorants for 10 other receptor neuron types were identified as (3Z)-hexenyl acetate, (+)-3-carene, trans-pinocarveol, trans-verbenol, vinylbenzaldehyde, 2-phenylethanol, methyl benzoate, alpha-caryophyllene and caryophyllene oxide, respectively. Most odorants were present in several host and non-host plant species, often in trace amounts. The specificity as well as the co-localization of particular neuron types so far recorded in both species showed similarities of the olfactory systems receiving plant odour information in these two species of heliothine moths.

New type of Sesiidae sex pheromone identified from the hornet moth Sesia apiformis.

Two components of the female-produced sex pheromone of the hornet moth, Sesia apiformis, were identified as (3Z,13Z)-octadeca-3,13-dien-1-ol (3Z,13Z-18:OH) and (2E,13Z)-octadeca-2,13-dienal (2E,13Z-18:Al), a pheromone structure new in Sesiidae. Pooled gland extracts showed the two major compounds in a proportion of ca. 2:3, while SPME-investigations on single calling females revealed a ratio of ca. 1:7. Although the single compounds were not attractive, a 2:3 mixture proved to be highly active towards males in field tests. Small amounts of (2E,13Z)-octadecadienol (2E,13Z-18:OH) were found in the sex pheromone gland of females, however, the biological significance of the compound remains unclear. Methyl sulfide was found to readily react with 2-alkenals, providing an effective new method for the characterization of this type of compound upon GC/MS. The derivatives, 1,1,3-tris(methylthio)alkanes, are the products of the addition of methyl sulfide to the double bond and the transformation of the carbonyl group into the corresponding bis(methylthio)acetal. The mass spectra of these compounds are characterized by diagnostic signals at m/z 107 and/or m/z 121. These fragments represent the first carbon unit or the first two carbon units of the derivative, respectively. The parent signal in the spectra of thiomethyl derivatives of 2-alkenals showing no other double bonds is represented by m/z M+ - 121, formed upon loss of the first two carbon units. By employing a solution of methyl sulfide in dimethyl sulfide, the double bond positions in 2E,13Z-18:Al could be fully characterized by GC/MS.

(-)-Germacrene D receptor neurones in three species of heliothine moths: structure-activity relationships.

Specificity of olfactory receptor neurones plays an important role in food and host preferences of a species, and may have become conserved or changed in the evolution of polyphagy and oligophagy. We have identified a major type of plant odour receptor neurones responding to the sesquiterpene germacrene D in three species of heliothine moths, the polyphagous Heliothis virescens and Helicoverpa armigera and the oligophagous Helicoverpa assulta. The neurones respond with high sensitivity and selectivity to (-)-germacrene D, as demonstrated by screening via gas chromatography with numerous mixtures of plant volatiles. Germacrene D was present in both host and non-host plants, but only in half of the tested species. The specificity of the neurones was similar in the three species, as shown by the "secondary" responses to a few other sesquiterpenes. The effect of (-)-germacrene D was about ten times stronger than that of the (+)-enantiomer, which again was about ten times stronger than that of (-)-alpha-ylangene. Weaker effects were obtained for (+)-beta-ylangene, (+)-alpha-copaene, beta-copaene and two unidentified sesquiterpenes. The structure-activity relationship shows that the important properties of (-)-germacrene D in activating the neurones are the ten-membered ring system and the three double bonds acting as electron-rich centres, in addition to the direction of the isopropyl-group responsible for the different effects of the germacrene D enantiomers.

Identification of plant odours activating receptor neurones in the weevil Pissodes notatus F. (Coleoptera, Curculionidae).

Plants release complex mixtures of volatiles important in the interaction with insects and other organisms. In the search for compounds that contribute to the perception of odour quality in the weevil Pissodes notatus, single olfactory receptor neurones on the antennae were screened for sensitivity to naturally produced plant volatiles by the use of gas chromatography linked to single cell recordings. We here present 60 olfactory neurones responding to 25 of the numerous compounds released by host and non-host plants. All the neurones show high selectivity and are classified into 12 distinct types. The two most abundant types respond to alpha-pinene, beta-pinene, and 3-carene ( n=17), and to isopinocamphone and pinocamphone ( n=17), respectively. Other neurone types respond to limonene ( n=9), beta-phellandrene ( n=3), and fenchone ( n=4). Responses to beta-caryophyllene ( n=1) and to ethanol ( n=4) are also shown. Except for two pairs, the neurone types do not show overlap of the molecular receptive range. The active compounds are present in the host, Pinus pinaster, as well as in non-hosts, supporting the idea that plant odour quality is mediated by the ratio of the compounds rather than specific odorants.

(-)-Germacrene D increases attraction and oviposition by the tobacco budworm moth Heliothis virescens.

The sesquiterpene germacrene D (GD) activates a major type of olfactory receptor neuron on the antennae of the heliothine moths. In Heliothis virescens females, 80% of the recordings have shown activity of one neuron type responding with high sensitivity and selectivity to GD. With the aim of determining the behavioural significance of this sesquiterpene, we have used a two-choice wind-tunnel to study the preference of mated H. virescens females for host plants with and without (-)-GD added. Tobacco plants containing dispensers with low release rate of (-)-GD had a greater attractiveness than tobacco plants without this substance. In addition, a significant increase of oviposition was found on the plants with (-)-GD.

Receptor neuron discrimination of the germacrene D enantiomers in the moth Helicoverpa armigera.

Plants release complex mixtures of volatiles, including chiral constituents. In the search for the biologically relevant plant odorants, gas chromatography linked to electrophysiological recordings from single receptor neurons has been employed. In heliothine moths, including the females of the Eurasian cotton bollworm moth Helicoverpa armigera, a major type of receptor neurons is identified, showing high sensitivity and selectivity for the sesquiterpene germacrene D. In the present study, gas chromatography with a chiral column linked to single cell recordings were performed. It was found that all germacrene D neurons belonged to one type; all responded to both enantiomers, but (-)-germacrene D had approximately 10 times stronger effect than (+)-germacrene D. Parallel dose-response curves for the two enantiomers were obtained by direct stimulations. The enantiomeric composition of germacrene D, which differed in six plant species and in different individuals of one species, was determined on the basis of the neuron responses. The results, showing the presence of one neuron type for receiving the information about germacrene D in the various plants, suggests that the two enantiomers mediate the same kind of information to the moth, but with different intensity.

Determination of hydrocarbons in old creosote contaminated soil using headspace solid phase microextraction and GC-MS.

Headspace solid phase microextraction (HS-SPME) has been used together with GC-MS to analyze organic substances directly in a soil, heavily contaminated with PAHs/creosote (approximately 300 mg/kg soil), from an old gaswork site in Stockholm, Sweden. The HS-SPME results, both qualitative and quantitative, were compared with traditional liquid extraction using ethyl acetate/hexane (20:80). It was shown that the concentrations determined with HS-SPME at 60 degrees C correlated well, for compounds containing up to two and three aromatic rings (naphthalenes, acenaphthene, acenaphthylene and fluorenes, while a lower concentration was obtained for phenanthrene, anthracene, fluoranthene and pyrene. The total concentrations for each compound determined with HS-SPME ranged from 2 to 25 microg/g soil. Quantification was done using standard addition of compounds directly to the soil samples. The bioavailable fraction of the compounds in the contaminated soil at 20 degrees C was analyzed using external calibration by spiking sterile uncontaminated sand (same texture and particle size as the contaminated soil but without a heavily sorbed organic fraction) with hydrocarbon standards in different concentrations. Storage of exposed fibers at 20 degrees C showed that analysis should be done within two days to make qualitative measurements and earlier (as soon as possible) for quantitative determinations.

Plant odor analysis of apple: antennal response of codling moth females to apple volatiles during phenological development.

Volatile compounds were collected from apple branches (Malus domestica) at different developmental stages, and the antennal response of codling moth females (Cydia pomonella) to these compounds was recorded by electroantennography coupled to gas chromatography. Presence of a range of terpenoid compounds, many of which had antennal activity, was characteristic for volatile collections from branches with leaves, and from small green apples. Nine compounds from branches with leaves and green fruit consistently elicited an antennal response: methyl salicylate, (E)-beta-farnesene, beta-caryophyllene, 4,8-dimethyl-1,3(E),7-nonatriene, (Z)3-hexenol, (Z,E)-alpha-farnesene, linalool, germacrene D, and (E,E)-alpha-farnesene. The bouquet emitted from flowering branches contained in addition several benzenoid compounds which were not found after bloom. Small green apples, which are the main target of codling moth oviposition during the first seasonal flight period, released very few esters. In comparison, fully grown apples released a large number of esters, but fewer terpenoids. The study of apple volatiles eliciting an antennal response, together with a survey of the seasonal change in the release of these compounds, is the first step toward the identification of volatiles mediating host-finding and oviposition in codling moth females.