Network Canvas: Key decisions in the design of an interviewer assisted network data collection software suite.

Self-reported social network analysis studies are often complex and burdensome, both during the interview process itself, and when conducting data management following the interview. Through funding obtained from the National Institute on Drug Abuse (NIDA/NIH), our team developed the Network Canvas suite of software - a set of complementary tools that are designed to simplify the collection and storage of complex social network data, with an emphasis on usability and accessibility across platforms and devices, and guided by the practical needs of researchers. The suite consists of three applications: Architect: an application for researchers to design and export interview protocols; Interviewer: a touch-optimized application for loading and administering interview protocols to study participants; and Server: an application for researchers to manage the interview deployment process and export their data for analysis. Together, they enable researchers with minimal technological expertise to access a complete research workflow, by building their own network interview protocols, deploying these protocols widely within a variety of contexts, and immediately attaining the resulting data from a secure central location. In this paper, we outline the critical decisions taken in developing this suite of tools for the network research community. We also describe the work which guides our decision-making, including prior experiences and key discovery events. We focus on key design choices, taken for theoretical, philosophical, and pragmatic reasons, and outline their strengths and limitations.

Novel use of PDMS tubing for in-soil capture of plant natural products.

The extraction of small lipophilic molecules (SLMs) in the soil-root interface that play a role in belowground ecological interactions between plants and insect herbivores was investigated. Polydimethylsiloxane (PDMS) microtubing has been shown to absorb root SLMs selectively in low-disturbance setups, where analytes were extracted from the polymer with methanol. This technique was adapted to isolate SLMs that diffuse in the vapour phase in soil and sand and under various experimental parameters, extracting with a plug of diethyl ether pushed through the length of the silicon tubing. Moisture level had a substrate-dependent effect on the recovery rate of analytes that were applied as synthetic blends of known belowground SLM semiochemicals in the media. Higher amounts of two selected SLMs, (E)-caryophyllene and (-)-thujopsene, were extracted from sand, and increased polymer and solvent volume, as well as sampling duration, resulted in more of these two SLMs recovered by extraction. It was also shown that PDMS tubes lose no extraction capacity after repeated use. The signature compound (E)-caryophyllene was successfully isolated from the rhizosphere of maize plants infested with Diabrotica v. virgifera larvae by extracting the silicon tubing with diethyl ether. Because the tubes are preconditioned to reduce the presence of contaminants, such extracts can be directly analysed by GC and GC-MS and used in electrophysiological and behavioural assays. After further modifications, non-invasive, in situ PDMS probes can be developed that extract SLMs from plant rhizosphere for the study of belowground chemical ecology processes.

Electrophysiologically and behaviourally active semiochemicals identified from bed bug refuge substrate.

Bed bugs are pests of public health importance due to their relentless biting habits that can lead to allergies, secondary infections and mental health issues. When not feeding on human blood bed bugs aggregate in refuges close to human hosts. This aggregation behaviour could be exploited to lure bed bugs into traps for surveillance, treatment efficacy monitoring and mass trapping efforts, if the responsible cues are identified. The aim of this study was to identify and quantify the bed bug aggregation pheromone. Volatile chemicals were collected from bed bug-exposed papers, which are known to induce aggregation behaviour, by air entrainment. This extract was tested for behavioural and electrophysiological activity using a still-air olfactometer and electroantennography, respectively. Coupled gas chromatography-electroantennography (GC-EAG) was used to screen the extract and the GC-EAG-active chemicals, benzaldehyde, hexanal, (E)-2-octenal, octanal, nonanal, decanal, heptanal, (R,S)-1-octen-3-ol, 3-carene, ß-phellandrene, (3E,5E)-octadien-2-one, (E)-2-nonenal, 2-decanone, dodecane, nonanoic acid, 2-(2-butoxyethoxy)ethyl acetate, (E)-2-undecanal and (S)-germacrene D, were identified by GC-mass spectrometry and quantified by GC. Synthetic blends, comprising 6, 16, and 18 compounds, at natural ratios, were then tested in the still-air olfactometer to determine behavioural activity. These aggregation chemicals can be manufactured into a lure that could be used to improve bed bug management.

Species-specific alterations in Anopheles mosquito olfactory responses caused by Plasmodium infection.

Mosquitoes infected with malaria parasites have demonstrated altered behaviour that may increase the probability of parasite transmission. Here, we examine the responses of the olfactory system in Plasmodium falciparum infected Anopheles gambiae, Plasmodium berghei infected Anopheles stephensi, and P. berghei infected An. gambiae. Infected and uninfected mosquitoes showed differential responses to compounds in human odour using electroantennography coupled with gas chromatography (GC-EAG), with 16 peaks triggering responses only in malaria-infected mosquitoes (at oocyst, sporozoite or both stages). A selection of key compounds were examined with EAG, and responses showed differences in the detection thresholds of infected and uninfected mosquitoes to compounds including lactic acid, tetradecanoic acid and benzothiazole, suggesting that the changes in sensitivity may be the reason for differential attraction and biting at the oocyst and sporozoite stages. Importantly, the different cross-species comparisons showed varying sensitivities to compounds, with P. falciparum infected An. gambiae differing from P. berghei infected An. stephensi, and P. berghei infected An. gambiae more similar to the P. berghei infected An. stephensi. These differences in sensitivity may reflect long-standing evolutionary relationships between specific Plasmodium and Anopheles species combinations. This highlights the importance of examining different species interactions in depth to fully understand the impact of malaria infection on mosquito olfactory behaviour.

Disruption of host-seeking behaviour by the salmon louse, Lepeophtheirus salmonis, using botanically derived repellents.

The potential for developing botanically derived natural products as novel feed-through repellents for disrupting settlement of the salmon louse, Lepeophtheirus salmonis (Caligidae) upon farmed Atlantic salmon, Salmo salar, was investigated using an established laboratory vertical Y-tube behavioural bioassay for assessing copepodid behaviour. Responses to artificial sea water conditioned with the odour of salmon, or to the known salmon-derived kairomone component, α-isophorone, in admixture with selected botanical materials previously known to interfere with invertebrate arthropod host location were recorded. Materials included oils extracted from garlic, Allium sativum (Amaryllidaceae), rosemary, Rosmarinus officinalis (Lamiaceae), lavender, Lavandula angustifolia (Lamiaceae), and bog myrtle, Myrica gale (Myricaceae), and individual components (diallyl sulphide and diallyl disulphide from garlic; allyl, propyl, butyl, 4-pentenyl and 2-phenylethyl isothiocyanate from plants in the Brassica genus). Removal of attraction to salmon-conditioned water (SCW) or α-isophorone was observed when listed materials were presented at extremely low parts per trillion (ppt), that is picograms per litre or 10-12 level. Significant masking of attraction to SCW was observed at a level of 10 ppt for diallyl disulphide and diallyl sulphide, and allyl isothiocyanate and butyl isothiocyanate. The potential of very low concentrations of masking compounds to disrupt Le. salmonis copepodid settlement on a host fish has been demonstrated in vitro.

Influence of Two Acyclic Homoterpenes (Tetranorterpenes) on the Foraging Behavior of Anthonomus grandis Boh.

Previous studies have shown that the boll weevil, Anthonomus grandis, is attracted to constitutive and conspecific herbivore-induced cotton volatiles, preferring the blend emitted by cotton at the reproductive over the vegetative stage. Moreover, this preference was paralleled by the release of the acyclic homoterpenes (tetranorterpenes) (E)-4,8-dimethyl-1,3,7-nonatriene (DMNT) and (E,E)-4,8,12-trimethyltrideca-1,3,7,11-tetraene (TMTT) in Delta Opal cotton being higher at the vegetative than at the reproductive stage. Here, we evaluated whether this difference in release of acyclic homoterpenes also occurred in other cotton varieties, and if boll weevils could recognize these compounds as indicators of a specific cotton phenological stage. Results showed that cotton genotypes CNPA TB-90, BRS-293 and Delta Opal all produced higher levels of DMNT and TMTT at the vegetative stage than at the reproductive stage and that these homoterpenes allowed for principal component analysis separation of volatiles produced by the two phenological stages. Electroantennograms confirmed boll weevil antennal responses to DMNT and TMTT. Behavioral assays, using Y-tube olfactometers, showed that adding synthetic homoterpenes to reproductive cotton volatiles (mimicking cotton at the vegetative stage in terms of homoterpene levels) resulted in reduced attraction to boll weevils compared to that to unmodified reproductive cotton. Weevils showed no preference when given a choice between plants at the vegetative stage and the vegetative stage-mimicked plant. Altogether, the results show that DMNT and TMTT are used by boll weevils to distinguish between cotton phenological stages.

Methods for Collection of Participant-aided Sociograms for the Study of Social, Sexual and Substance-using Networks Among Young Men Who Have Sex with Men.

In this study, we adapted and tested a participant-aided sociogram approach for the study of the social, sexual, and substance use networks of young men who have sex with men (YMSM); a population of increasing and disproportionate risk of HIV infection. We used a combination of two interviewer-administered procedures: completion of a pre-numbered list form to enumerate alters and to capture alter attributes; and a participant-aided sociogram to capture respondent report of interactions between alters on an erasable whiteboard. We followed the collection of alter interactions via the sociogram with a traditional matrix-based tie elicitation approach for a sub-sample of respondents for comparison purposes. Digital photographs of each network drawn on the whiteboard serve as the raw data for entry into a database in which group interactions are stored. Visual feedback of the network was created at the point of data entry, using NetDraw network visualization software for comparison to the network structure elicited via the sociogram. In a sample of 175 YMSM, we found this approach to be feasible and reliable, with high rates of participation among those eligible for the study and substantial agreement between the participant-aided sociogram in comparison to a traditional matrix-based approach. We believe that key strengths of this approach are the engagement and maintenance of participant attention and reduction of participant burden for alter tie elicitation. A key weakness is the challenge of entry of interview-based list form and sociogram data into the database. Our experience suggests that this approach to data collection is feasible and particularly appropriate for an adolescent and young adult population. This builds on and advances visualization-based approaches to social network data collection.

Identification of pheromone components and their binding affinity to the odorant binding protein CcapOBP83a-2 of the Mediterranean fruit fly, Ceratitis capitata.

The Mediterranean fruit fly (or medfly), Ceratitis capitata (Wiedemann; Diptera: Tephritidae), is a serious pest of agriculture worldwide, displaying a very wide larval host range with more than 250 different species of fruit and vegetables. Olfaction plays a key role in the invasive potential of this species. Unfortunately, the pheromone communication system of the medfly is complex and still not well established. In this study, we report the isolation of chemicals emitted by sexually mature individuals during the "calling" period and the electrophysiological responses that these compounds elicit on the antennae of male and female flies. Fifteen compounds with electrophysiological activity were isolated and identified in male emissions by gas chromatography coupled to electroantennography (GC-EAG). Within the group of 15 identified compounds, 11 elicited a response in antennae of both sexes, whilst 4 elicited a response only in female antennae. The binding affinity of these compounds, plus 4 additional compounds known to be behaviourally active from other studies, was measured using C. capitata OBP, CcapOBP83a-2. This OBP has a high homology to Drosophila melanogaster OBPs OS-E and OS-F, which are associated with trichoid sensilla and co-expressed with the well-studied Drosophila pheromone binding protein LUSH. The results provide evidence of involvement of CcapOBP83a-2 in the medfly's odorant perception and its wider specificity for (E,E)-α-farnesene, one of the five major compounds in medfly male pheromone emission. This represents the first step in the clarification of the C. capitata and pheromone reception pathway, and a starting point for further studies aimed towards the creation of new powerful attractants or repellents applicable in the actual control strategies.

Identification and Expression Profile of Two Putative Odorant-Binding Proteins from the Neotropical Brown Stink Bug, Euschistus heros (Fabricius) (Hemiptera: Pentatomidae).

This work presents the first identification of putative odorant-binding proteins (OBPs) from a member of the Pentatomidae, i.e. the brown stink bug Euschistus heros (Fabricius), an important pest of soybean in Brazil. Antennae from both sexes of E. heros adults (12 days old and unmated) were used to construct a cDNA library, from which two transcripts encoding putative E. heros OBPs (EherOBPs) were identified. The expression levels of EherOBP1 and EherOBP2 were found to be higher in male antennae than in female and there was difference in expression in legs, wings, and abdomens of the two sexes. The histolocalization of EherOBP1 and EherOBP2 transcripts in antennae also showed a sexual dimorphism in the chemoreception system, with different expression sites in the antennal segments between males and females, occurring predominantly at the base of the sensillum. The implications of these findings for stink bug chemoreception are discussed.

Evaluation of a nonanal-trimethylamine lure for collection of Culex quinquefasciatus (Diptera: Culicidae) in gravid traps.

Gravid traps are useful tools for monitoring vector-borne pathogens in mosquitoes, particularly for those pathogens transmitted by Culex quinquefasciatus Say. One of the primary challenges in the use of gravid traps is the necessity of the inclusion of an oviposition attractant, usually an infusion of organic material, which changes in attractiveness over time. However, a standardized lure, using nonanal and trimethylamine (N + TMA), has been developed and is commercially available. The N + TMA lure was tested against grass infusion and tap water in Tanzania, where Cx. quinquefasciatus is a vector of lymphatic filariasis. Traps baited with grass infusion collected significantly more mosquitoes than N + TMA-baited traps, which collected significantly more than traps baited with tap water. The advantages and disadvantages of the standardized lure are discussed.

Tracking bed bugs (Cimex lectularius): a study of the effect of physiological and extrinsic factors on the response to bed bug-derived volatiles.

The common bed bug, Cimex lectularius, feeds on the blood of mammal and bird hosts, and is a pest of global importance. Semiochemicals are chemicals involved in animal communication that may affect behaviour and/or physiology. Attractive semiochemicals that play a role in mediating bed bug behaviour could be exploited for the development of a highly effective novel monitoring device. Tracking software was used to record the response of bed bugs to volatiles from paper previously exposed to conspecific bugs in a still-air olfactometer illuminated by infrared lights, through a variety of activity variables. The effect of time of day as an extrinsic factor, and sex, stage, mating status and nutritional status as physiological factors on the response of bed bugs to the volatiles was examined. Bed bugs of both sexes and all stages responded to the volatiles from bed bug-exposed papers, showing significant attraction and orientation towards the volatile source whether they were starved or engorged. Confirmation that the physiological factors examined do not affect the response of bed bugs to the volatiles from bed bug-exposed papers provides evidence that these bed bug-derived volatiles contain aggregation cues, as semiochemicals that promote aggregation should by definition be detected by both sexes and all life stages. A device baited with such semiochemicals could play a major role in limiting the impact of the current bed bug resurgence by enabling timely detection of infestations, along with quantitative evaluation of control and effective surveillance of the geographical distribution of the pest species.

Semiochemicals from herbivory induced cotton plants enhance the foraging behavior of the cotton boll weevil, Anthonomus grandis.

The boll weevil, Anthonomus grandis, has been monitored through deployment of traps baited with aggregation pheromone components. However, field studies have shown that the number of insects caught in these traps is significantly reduced during cotton squaring, suggesting that volatiles produced by plants at this phenological stage may be involved in attraction. Here, we evaluated the chemical profile of volatile organic compounds (VOCs) emitted by undamaged or damaged cotton plants at different phenological stages, under different infestation conditions, and determined the attractiveness of these VOCs to adults of A. grandis. In addition, we investigated whether or not VOCs released by cotton plants enhanced the attractiveness of the aggregation pheromone emitted by male boll weevils. Behavioral responses of A. grandis to VOCs from conspecific-damaged, heterospecific-damaged (Spodoptera frugiperda and Euschistus heros) and undamaged cotton plants, at different phenological stages, were assessed in Y-tube olfactometers. The results showed that volatiles emitted from reproductive cotton plants damaged by conspecifics were attractive to adults boll weevils, whereas volatiles induced by heterospecific herbivores were not as attractive. Additionally, addition of boll weevil-induced volatiles from reproductive cotton plants to aggregation pheromone gave increased attraction, relative to the pheromone alone. The VOC profiles of undamaged and mechanically damaged cotton plants, in both phenological stages, were not different. Chemical analysis showed that cotton plants produced qualitatively similar volatile profiles regardless of damage type, but the quantities produced differed according to the plant's phenological stage and the herbivore species. Notably, vegetative cotton plants released higher amounts of VOCs compared to reproductive plants. At both stages, the highest rate of VOC release was observed in A. grandis-damaged plants. Results show that A. grandis uses conspecific herbivore-induced volatiles in host location, and that homoterpene compounds, such as (E)-4,8-dimethylnona-1,3,7-triene and (E,E)-4,8,12-trimethyltrideca-1,3,7,11-tetraene and the monoterpene (E)-ocimene, may be involved in preference for host plants at the reproductive stage.

Sex pheromone communication in two sympatric neotropical stink bug species Chinavia ubica and Chinavia impicticornis.

Chinavia and Nezara spp. stink bugs (Heteroptera: Pentatomidae) include over100 species, with highest diversity in Afrotropical and Neotropical regions. Species thus far studied in these genera utilize trans-(Z)-(4 S)-bisabolene epoxide (BE) and cis-(Z)-(4 S)-BE as major sex pheromone components, with species specificity ensured by different ratios of the two compounds. Gas chromatography (GC) and coupled GC-mass spectrometry (GC-MS) analyses of a volatiles from C. ubica males revealed the presence of two BE isomers in approximately a 90:10 ratio, which were shown by microprobe (1) H NMR to be cis-(Z)-BE and trans-(Z)-BE isomers, respectively. Analyses of volatiles from C. impicticornis males suggested the presence of a single isomer, trans-(Z)-BE, in high purity (>90 %). The absolute configurations of the isomers produced by C. ubica and C. impicticornis were determined using chiral GC analysis (ß-DEX column). Oxidative microchemistry of synthetic standards of cis-(Z)-(4 S)-BE and trans-(Z)-(4R)-BE, and volatiles from male of C. ubica, revealed the absolute stereochemistry of the cis-(Z)-BE to be (1R,2 S,4 S) [cis-(Z)-(4 S) for short]. Similarly, analyses of trans-(Z)-(4 S)-BE and cis-(Z)-(4R)-BE standards, and volatiles from males of C. ubica and C. impicticornis, revealed the absolute stereochemistry of the trans-(Z)-BE to be (1 S,2R,4 S) [trans-(Z)-(4 S) for short]. Olfactometer bioassays with synthetic BEs confirmed attraction of female C. ubica and C. impicticornis to conspecific synthetic pheromone, but not to heterospecific synthetic pheromone. Chinavia impicticornis appeared not to discriminate behaviorally between the conspecific pheromone and its enantiomer. Coupled GC-electroantennography with antennae from females suggested that C. ubica and C. impicticornis possess olfactory receptors for both cis-(Z)-(4 S)-BE and trans-(Z)-(4 S)-BE. The results in this study confirm that C. ubica and C. impicticornis, as for other Chinavia and Nezara spp., utilize cis-(Z)-(4 S)-BE and trans-(Z)-(4 S)-BE as sex pheromone components, with different ratios guaranteeing species specificity. Furthermore, the results suggest that the absolute stereochemistry of BEs may be less important for conspecific recognition than the relative stereochemistry between the epoxide group and the alkyl substituent on the bisabolene ring.

Interference in foraging behaviour of European and American house dust mites Dermatophagoides pteronyssinus and Dermatophagoides farinae (Acari: Pyroglyphidae) by catmint, Nepeta cataria (Lamiaceae).

The European and American house dust mites, Dermatophagoides pteronyssinus and D. farinae, have a huge impact upon human health worldwide due to being the most important indoor trigger of atopic diseases such as asthma, rhinitis and atopic dermatitis. Preceding studies have shown that the behavioural response of house dust mites towards volatile chemicals from food sources can be assessed using a Y-tube olfactometer assay. In the current study, we used this assay to investigate, for the first time, the ability of the essential oil of the catmint plant, Nepeta cataria (Lamiaceae), known to repel other ectoparasites affecting human and animal health, to interfere with the attraction of D. pteronyssinus and D. farinae towards a standard food source (fish flakes). Two distinct chemotypes (A and B), enriched in the iridoid compounds (4aS,7S,7aR)-nepetalactone and (4aS,7S,7aS)-nepetalactone, and the sesquiterpene (E)-(1R,9S)-caryophyllene, were used. Initial assays with a hexane extract of fish flakes (FF extract) confirmed attraction of mites to this positive control (P < 0.001 and P < 0.05 for D. pteronyssinus and D. farinae respectively), but when presented in combination with either N. cataria chemotype, tested across a range of doses (10, 1, 0.1 and 0.01 µg), decreasing attraction of mites to their food source was observed as the dose augmented. Our study shows that N. cataria, enriched in iridoid nepetalactones and (E)-(1R,9S)-caryophyllene, exhibits potent repellent activity for house dust mites, and has the potential for deployment in control programmes based on interference with normal house dust mite behaviour.

Collection of Culicoides (Diptera: Ceratopogonidae) using CO2 and enantiomers of 1-octen-3-ol in the United Kingdom.

The host kairomones carbon dioxide (CO2) and 1-octen-3-ol elicit a host seeking response in a wide range of haematophagous Diptera. This study investigates the response of Culicoides biting midges (Diptera: Ceratopogonidae) to these cues using field-based experiments at two sites in the United Kingdom with very different species complements. Traps used for surveillance (miniature CDC model 512) and control (Mosquito Magnet Pro) were modified to release ratios of (R)- and (S)-1-octen-3-ol enantiomers in combination with CO2 and, in the case of the latter trap type, a thermal cue. Abundance and species diversity were then compared between these treatments and against collections made using a trap with a CO2 lure only, in a Latin square design. In both habitats, results demonstrated that semiochemical lures containing a high proportion of the (R)-enantiomer consistently attracted a greater abundance of host-seeking Culicoides females than any other treatment. Culicoides collected using an optimal stimulus of 500 ml/min CO2 combined with 4.1 mg/h (R)-1-octen-3-ol were then compared with those collected on sheep through the use of a drop trap. While preliminary in nature, this trial indicated Culicoides species complements are similar between collections made using the drop trap in comparison to the semiochemical-baited CDC trap, and that there are advantages in using (R)-1-octen-3-ol.

Enantiomeric selectivity in behavioural and electrophysiological responses of Aedes aegypti and Culex quinquefasciatus mosquitoes.

1-Octen-3-ol is a kairomone for many haematophagous insects including mosquitoes. Numerous studies have examined the effects of racemic 1-octen-3-ol; however, few studies have investigated the role of individual enantiomers in relation to mosquito attraction. In the present study, we investigated the behavioural and electrophysiological responses of two mosquito species, Aedes aegypti and Culex quinquefasciatus, to individual enantiomers and mixtures of 1-octen-3-ol, employing a laboratory Y-tube olfactometer and single sensillum recordings. The olfactory receptor neurons of both Ae. aegypti and Cx. quinquefasciatus had a significantly higher response to the (R)-1-octen-3-ol enantiomer compared to the (S)-1-octen-3-ol enantiomer at 10-9 g µl-1 to 10-6 g µl-1. Behaviourally, Ae. aegypti was more responsive to the (R)-1-octen-3-ol enantiomer, showing an increase in flight activity and relative attraction compared to Cx. quinquefasciatus. The (R)-1-octen-3-ol enantiomer caused an increase in activation for Cx. quinquefasciatus. However, the most notable effect was from an (R:S)-1-octen-3-ol mixture (84:16) that caused significantly more mosquitoes to sustain their flight and reach the capture chambers (demonstrated by a reduced non-sustained flight activity), suggesting that it may have a behaviourally excitatory effect. For Cx. quinquefasciatus, a reduced relative attraction response was also observed for all treatments containing the (R)-1-octen-3-ol enantiomer, either on its own or as part of a mixture, but not with the (S)-1-octen-3-ol enantiomer. This is the first time enantiomeric selectivity has been shown for Ae. aegypti using electrophysiology in vivo. The implications of these results for exploitation in mosquito traps are discussed.

A bioassay for studying behavioural responses of the common bed bug, Cimex lectularius (Hemiptera: Cimicidae) to bed bug-derived volatiles.

The common bed bug, Cimex lectularius (Hemiptera: Cimicidae), has recently re-emerged in increasing numbers, distribution and intensity of infestation in many countries. Current control relies on the application of residual pesticides; but, due to the development of insecticide resistance, there is a need for new tools and techniques. Semiochemicals (behaviour and physiology modifying chemicals) could be exploited for management of bed bugs. However, in order to identify semiochemicals that can be utilised in monitoring or control, a suitable olfactometer is needed that enables the study of the responses of bed bugs to volatile chemicals. Previous studies have used olfactometers that do not separate olfactory responses from responses to physical contact. In this study, a still-air olfactometer was used to measure behavioural responses to different bed bug-derived volatiles presented in an odour pot. Bed bugs were significantly more likely to visit the area above the odour pot first, and more frequently, in the presence of volatiles from bed bug-exposed paper but not in the presence of volatiles from conspecific bed bugs. Bed bug activity was found to be dependent on the presence of the volatiles from bed bug-exposed paper, the time during the scotophase and the sex of the insect being tested. The still-air olfactometer could be used to test putative semiochemicals, which would allow an understanding of their behavioural role in bed bug ecology. Ultimately, this could lead to the identification of new semiochemical tools for bed bug monitoring and control.

Identification of neryl formate as the airborne aggregation pheromone for the American house dust mite and the European house dust mite (Acari: Epidermoptidae).

The American house dust mite, Dermatophagoidesfarinae Hughes, and European house dust mite, Dermatophagoides pteronyssinus Trouessart, are major pests of medical importance throughout the developed world, causing atopic diseases such as asthma, rhinitis, and atopic dermatitis. Previous studies in our laboratory have shown that the behavioral responses of house dust mites toward volatiles from food sources could be assessed using a Y-tube olfactometer assay. The current study used this Y-tube assay to investigate house dust mite pheromones. A hexane extract of D.farinae, along with fractions of the extract prepared by microscale liquid chromatography over Florisil, were tested for behavioral activity. One of the chromatographic fractions was shown to be significantly attractive (P < 0.05) for D. farinae, compared with a solvent control. Coupled gas chromatography-mass spectrometry analysis of this behaviorally active fraction indicated that neryl or geranyl formate was the major component. Peak enhancement by gas chromatography, using authentic samples of the neryl and geranyl isomers prepared in high purity by chemical synthesis, confirmed the identity of the major peak as neryl formate. In Y-tube assays, male and female D. farinae and D. pteronyssinus both were significantly attracted to synthetic neryl formate at doses of 100 and 10 ng, respectively (P < 0.05). No significant differences were found for D. farinae and D. pteronyssinus when synthetic neryl formate and house dust mite extracts containing natural neryl formate were tested at the same level. Dynamic headspace collection of D. farinae and D. pteronyssinus colonies showed that neryl formate was released as a volatile organic compound by both species. Our study shows that neryl formate is an aggregation pheromone for D. farinae and D. pteronyssinus, and has the potential to be used as part of a novel lure-and-kill system for house dust mite control.

Novel use of stir bar sorptive extraction (SBSE) as a tool for isolation of oviposition site attractants for gravid Culex quinquefasciatus.

Mosquitoes such as Culex quinquefasciatus Say (Diptera: Culicidae) are important vectors of organisms that cause disease in humans. Research into the development of effective standardized odour baits for blood-fed females (oviposition attractants), to enable entomological monitoring of vector populations, is hampered by complex protocols for extraction of physiologically active volatile chemicals from natural breeding site water samples, which have produced inconsistent results. Air entrainment and solvent extraction are technically demanding methods and are impractical for use in resource poor environments where mosquito-borne disease is most prevalent. This study reports the first use of a simple, robust extraction technique, stir bar sorptive extraction (SBSE), to extract behaviourally active small lipophilic molecules (SLMs) present in water samples collected from Cx. quinquefasciatus breeding sites in Tanzania. Extracts from a pit latrine and from a cess pool breeding site attracted more gravid Cx. quinquefasciatus in pair choice bioassays than control extracts, and coupled gas chromatography-electroantennography (GC-EAG) allowed tentative identification of 15 electrophysiologically active chemicals, including the known oviposition attractant, skatole (3-methylindole). Here, we have demonstrated, using simple pair choice bioassays in controlled laboratory conditions, that SBSE is effective for the extraction of behaviourally and electrophysiologically active semiochemicals from mosquito breeding site waters. Further research is required to confirm that SBSE is an appropriate technique for use in field surveys in the search for oviposition cues for Cx. quinquefasciatus.