Pollination by nectar-foraging pompilid wasps: a new specialized pollination strategy for the Australian flora.

The Pompilidae is a cosmopolitan and diverse group of wasps, which commonly feed on nectar. However, pollination systems specialized on pompilids have not been documented in detail outside of southern Africa. Here, we studied Caladenia drummondii (Orchidaceae) where, based on floral traits and preliminary field observations, we predicted pollination by sexual deception of male pompilid wasps. Detailed pollinator observations were undertaken using floral baiting experiments at sites spanning 375 km. Following evidence for nectar on some flowers of C. drummondii, the sugar content on the labellum was analysed by GC-MS. Floral spectral reflectance was measured and compared with Caladenia using other pollination strategies. Males of a single species of pompilid wasp (Calopompilus sp.) were the only visitors capable of pollinating C. drummondii. Attempts to feed from the surface of the labellum were frequent and were associated with removal and deposition of pollinia. GC-MS analysis revealed larger quantities of sugar on the labellum than reported in other Caladenia species. While no sexual or courtship behaviour was observed, the zig-zag and circling flight on approach to the flower is suggestive of odour-based attraction. Floral spectral reflectance was similar to sexually deceptive Caladenia. This study represents the first confirmation of a specialized pompilid pollination system outside of Africa. Although pollination occurs during nectar-foraging, long-distance sexual attraction cannot be ruled out as an explanation for the exclusive male visitation. The similarity in floral spectral reflectance to other Caladenia indicates colour may not impose a constraint on the evolution of pollination by pompilids.

A specialised pollination system using nectar-seeking thynnine wasps in Caladenia nobilis (Orchidaceae).

Caladenia is a diverse Australian genus that is exceptional among orchids in having both species pollinated by food-seeking and sexually deceived insects. Here, we investigated the pollination of Caladenia nobilis, a species predicted to be food-deceptive due to its large, cream-coloured and apparently nectarless flowers. Pollinator observations were made using experimental clumps of flowers. Measurements of floral colour were undertaken with a spectrometer, nectar was tested using GC-MS, and reproductive success was quantified for 2 years. While C. nobilis attracted nine species of insect, only males of the thynnine wasp Rhagigaster discrepans exhibited the correct size and behaviour to remove and deposit pollen. Male R. discrepans attempted to feed from the surface of the labellum, often crawling to multiple flowers, but showed no evidence of sexual attraction. Most flowers produced little or no nectar, although some may provide enough sucrose to act as a meagre reward to pollinators. Floral colouration was similar to a related Caladenia species pollinated by sexual deception, although the sexually deceptive species had a dull-red labellum. Reproductive success was generally low and highly variable between sites and years. In addition to most visitors being of inappropriate size for pollinia removal, the lack of response to the orchid by several co-occurring species of thynnine wasp suggests filtering of potential pollinators at the attraction phase. Our discovery of a pollination strategy that may be intermediate between food deception and food reward raises the question, how many putatively rewardless orchids actually produce meagre amounts of nectar?

Comparison of a 2D iPad application and 3D body scanner to air displacement plethysmography for measurement of body fat percentage.

BACKGROUND: Novel and innovative imaging methods that rapidly estimate body fat percentage (%BF) are publicly available, yet little is known about their accuracy. The present study evaluated the test-retest reliability of a two-dimensional iPad (Apple, Inc., Cupertino, CA, USA) application (2D APP) and a three-dimensional body scanner (3D SCAN) for estimating %BF and compared both imaging methods with air displacement plethysmography (Bod Pod; Cosmed USA, Inc., Concord, CA, USA). METHODS: Seventy-nine adults (37 female, 42 male) varying widely in age [mean (SD), range] [32.9 (12.4), 18-65 years] and body mass index [25.0 (4.9), 18.2-41.8 kg m-2 ] were measured with the Bod Pod and twice with the 3D SCAN and the 2D APP in a repeated-measures design. RESULTS: Test-retest reliability was excellent for both the 2D APP (intraclass correlation = 0.993) and the 3D SCAN (intraclass correlation = 0.993) with the SEM <1% BF for both methods. Although the three methods were highly correlated with each other (r = 0.857-0.923), the mean %BF estimations were significantly different (P = 0.001). The 2D APP [19.9 (8.2)%BF] underestimated the Bod Pod value [21.9 (9.4)%BF] and the 3D SCAN [24.0 (6.8)%BF] overestimated. Additionally, the SE of estimate and total error exceeded 4% BF for both 2D APP and 3D SCAN, and both methods tended to overestimate lean participants and underestimate fat participants. CONCLUSIONS: Although highly reliable, neither the 2D APP, nor the 3D SCAN provided valid estimates of %BFBod Pod .

Volatiles from Merino fleece evoke antennal and behavioural responses in the Australian sheep blow fly Lucilia cuprina.

To identify flystrike-related volatile compounds in wool from Merino sheep, the attractiveness of wool to Lucilia cuprina Wiedmann (Diptera: Calliphoridae) was examined. First, a selection of wool samples guided by previous knowledge of sheep lines, predicted to be more susceptible or more resistant to flystrike, was tested. The attractiveness of the 10 samples selected was not associated with field susceptibility: two samples from the more resistant line were identified as most attractive and two samples from the more susceptible line were identified as least attractive, based on the behavioural assays with gravid flies. Comparison of the headspace volatiles of these samples, using solid phase microextraction and gas chromatography-mass spectrometry-electroantennographic detection, revealed octanal and nonanal to be present in the attractive wool samples that elicited responses from the fly antenna. Furthermore, the two compounds were not present in wool that was least attractive to L. cuprina. In laboratory bioassays, octanal and nonanal evoked antennal and behavioural responses in gravid L. cuprina, thus confirming their potential role as semiochemicals responsible for attracting L. cuprina to Merino sheep.

Mismatch in the distribution of floral ecotypes and pollinators: insights into the evolution of sexually deceptive orchids.

Plants are predicted to show floral adaptation to geographic variation in the most effective pollinator, potentially leading to reproductive isolation and genetic divergence. Many sexually deceptive orchids attract just a single pollinator species, limiting opportunities to experimentally investigate pollinator switching. Here, we investigate Drakaea concolor, which attracts two pollinator species. Using pollinator choice tests, we detected two morphologically similar ecotypes within D. concolor. The common ecotype only attracted Zaspilothynnus gilesi, whereas the rare ecotype also attracted an undescribed species of Pogonothynnus. The rare ecotype occurred at populations nested within the distribution of the common ecotype, with no evidence of ecotypes occurring sympatrically. Surveying for pollinators at over 100 sites revealed that ecotype identity was not correlated with wasp availability, with most orchid populations only attracting the rare Z. gilesi. Using microsatellite markers, genetic differentiation among populations was very low (GST = 0.011) regardless of ecotype, suggestive of frequent gene flow. Taken together, these results may indicate that the ability to attract Pogonothynnus has evolved recently, but this ecotype is yet to spread. The nested distribution of ecotypes, rather than the more typical formation of ecotypes in allopatry, illustrates that in sexually deceptive orchids, pollinator switching could occur throughout a species' range, resulting from multiple potentially suitable but unexploited pollinators occurring in sympatry. This unusual case of sympatric pollinators highlights D. concolor as a promising study system for further understanding the process of pollinator switching from ecological, chemical and genetic perspectives.

Infrequent attention to dietary and physical activity behaviours in conversations in Swedish child health services.

AIM: To investigate conversations between nurses and parents in Swedish child health services and to assess to what extent attention is directed towards dietary and physical activity behaviours in children. METHODS: Twenty-three nurses audio-recorded one session each. Recordings were assessed and topics were classified according to predetermined categories. RESULTS: The three most frequent topics of conversation concerned physical examinations of the child (30% of session time), talking to the child to establish or maintain contact and interest (15%), and development of language skills (12%). Dietary habits came on fourth place (10%), and physical activity ranked 14 (4%). CONCLUSION: Attention to dietary and physical activity behaviours in children is infrequent in Swedish child health services. Concern is raised about the efficacy of prevention efforts against childhood obesity.

Identification and electrophysiological studies of (4S,5S)-5-hydroxy-4-methyl-3-heptanone and 4-methyl-3,5-heptanedione in male lucerne weevils.

An investigation to identify a sex or aggregation pheromone of Sitona discoideus Gyllenhål (Coleoptera: Curculionidae) is presented. Antenna flicking and attraction behaviors evoked by conspecifics of both sexes were recorded in arena bioassays, where attraction of females to males was observed. Air entrainment of both males and females was conducted in separate chambers. Gas chromatographic-mass spectrometric analysis of headspace volatiles revealed that two male-specific compounds, 4-methyl-3,5-heptanedione (major) and (4S,5S)-5-hydroxy-4-methyl-3-heptanone (minor), were emitted during the autumnal post-aestivatory flight period. The stereoisomers of the minor component were separated by enantioselective gas chromatography and their absolute configurations assigned by NMR (diastereomers) and the known preference of enantioselective transesterification reactions catalyzed by Candida antarctica lipase B. Electroantennogram and single sensillum recording studies indicate that 4-methyl-3,5-heptanedione as well as all individual stereoisomers of 5-hydroxy-4-methyl-3-heptanone are detected by the antennae of male and female S. discoideus. Further, single sensillum recordings suggest that both sexes of S. discoideus have specialized olfactory receptor neurons (ORNs) for detecting 4-methyl-3,5-heptanedione and different populations of stereoselective ORNs for detecting the stereoisomers of 5-hydroxy-4-methyl-3-heptanone. Some of these stereoselective ORNs appear to be sex-specific in S. discoideus.

Structure elucidation and synthesis of dioxolanes emitted by two Triatoma species (Hemiptera: Reduviidae).

Volatiles from the metasternal glands of two species of true bugs of the Triatominae subfamily, Triatoma brasiliensis and Triatoma infestans, were analyzed by SPME-GC/MS. Two sets of new natural products were found: (4S,5S)- and (4R,5R)-2,2,4-triethyl-5-methyl-1,3-dioxolane (1) (major component) and (4S*,5S*)-2,4-diethyl-2,5-dimethyl-1,3-dioxolane (2) (trace component), (2R/S,4S,5S)- as well as (2R/S,4R,5R)-4-ethyl-5-methyl-2-(1-methylethyl)-1,3-dioxolane (3) (minor component), (2R/S,4S*,5S*)-4-ethyl-5-methyl-2-(1-methylpropyl)-1,3-dioxolane (4) (trace component), and (2R/S,4S*,5S*)-4-ethyl-5-methyl-2-(2-methylpropyl)-1,3-dioxolane (5) (trace component). Syntheses of optically active 1 and 3 were carried out by reacting pure enantiomers of 2,3-pentanediol with 3-pentanone or 2-methylpropanal. The preparation of pure stereoisomers of 2,3-pentanediol involved a novel key step for the synthesis of secondary alcohols: the reduction of a carboxylic ester by means of DIBAH and in situ alkylation of the intermediate by Grignard reaction at low temperature. Starting from the pure enantiomers of methyl lactate, all four stereoisomers of 2,3-pentanediol were synthesized and transformed to the corresponding isomers of 1 and 2. Relative configurations of the natural products and enantiomeric compositions of naturally occurring 1 and 2 were determined by comparison of their mass spectra and gas chromatographic retention times (co-injection) with those of authentic reference samples.

(4S,5S)-2,2,4-Triethyl-5-methyl-1,3-dioxolane: a new volatile released by a triatomine bug.

Adults of the triatomine bug Triatoma brasiliensis release 2,2,4-triethyl-5-methyl-1,3-dioxolane (1) as a mixture of the (4S,5S)- and (4R,5R)-enantiomers in a ratio of 4:1. Among the volatile acetals identified from insects so far, this is the first example resulting from an intermolecular condensation of a carbonyl moiety and a diol substructure.

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.

Central cholinergic effects of tricyclic antidepressants in mouse.

The apparent rate of acetylcholine (ACh) turnover and levels of ACh and choline (Ch) in whole mouse brain were examined 30 min following i.p. injection of 25 mg/kg of three tricyclic antidepressants (TAD's): amitriptyline, chlorimipramine, and imipramine. The effects of these agents on high affinity choline uptake (HACU), a rate-limiting, regulatory step in ACh synthesis, were also examined. All three TAD's inhibited ACh turnover (by 27-40%), increased Ch levels (by 33-37%), and inhibited HACU (IC50's from 1.7 - 6.8 X 10(-5) M). Two of the three drugs produced significant decreases in ACh levels. It is concluded that these agents possess anticholinergic activity which is independent of their previously demonstrated muscarinic receptor blocking capabilities. As substantial evidence has accumulated linking increased cholinergic function with depression, it is speculated that this additional anticholinergic activity may contribute to the clinical efficacy of the TAD's.