Aedes aegypti oviposition-sites choice under semi-field conditions.

Vector control is still the recommended approach to avoid arbovirus outbreaks. Herein, we investigate oviposition preferences of Aedes aegypti (Diptera: Culicidae) females under a semi-field structure Rio de Janeiro, Brazil. For that, in Experiment 1, we used two settings: 'Single items', which included as containers drain, beer bottle, bucket, car tyre, water tank, and a potted Peace Lily (Spathiphyllum wallisii) in a saucer with water, or 'Multiple containers', as an urban simulation, in which one drain, two additional beer bottles, and an extra plant pot saucer were added. Experiment 2 (sensory cues) used five variations of potted plant, each one varying in the range of sensory cues known to attract gravid females to oviposition containers. Our results indicate that gravid Ae. aegypti prefer to oviposit close to the ground and in open water containers with organic compounds from plant watering. Domestic large artificial containers containing tap water received significantly fewer eggs, except for the car tyre, which exhibited as many eggs as the potted plant. We also show that visual (potted plant shape) and olfactory clues (odour of the plant or from water containing organic matter) were equally attractive separately as were these stimuli together.

Components of the Female Sex Pheromone of the Newly-Described Canola Flower Midge, Contarinia brassicola.

The canola flower midge, Contarinia brassicola Sinclair (Diptera: Cecidomyiidae), is a newly-described species that induces galls on canola, Brassica napus Linnaeus and Brassica rapa Linnaeus (Brassicaceae). Identification of the sex pheromone of C. brassicola is essential to developing monitoring tools to elucidate the geographic range and hosts of this new pest, and the extent to which it threatens the $30 billion Canadian canola industry. The aim of this study was to identify and synthesize the female-produced sex pheromone of C. brassicola and demonstrate its effectiveness in attracting males to traps in the field. Two peaks were identified through GC-EAG analysis of female-produced volatiles which elicited electrophysiological responses in male antennae. These peaks were initially characterized through GC-MS and synthesis as 2,7-diacetoxynonane (major component) and 2-acetoxynonane (minor component), and the racemic compounds elicited EAG responses in male antennae. All four stereoisomers of 2,7-diacetoxynonane were synthesized and the naturally-produced compound was shown to be primarily the (2R,7S)-isomer by analysis on an enantioselective GC column, with a small amount of (2R,7R)-2,7-diacetoxynonane also present. The configuration of the minor component could not be determined because of the small amount present, but this was assumed to be (2R)-2-acetoxynonane by comparison with the configuration of the other two components. In field trials, none of the four stereoisomers of 2,7-diacetoxynonane, presented individually or as a racemic mixture, was attractive to male C. brassicola. However, dispensers loaded with a 10 µg:1 µg blend of (2R,7S)- and (2R,7R)-2,7-diacetoxynonane caught large numbers of male C. brassicola and significantly more than other blends tested. The addition of 0.5 µg of (2R)-2-acetoxynonane to this blend further increased the number of males caught. In future work, we will seek to identify the optimum trapping protocol for the application of the pheromone in monitoring and surveillance.

Hero Turned Villain: Identification of Components of the Sex Pheromone of the Tomato Bug, Nesidiocoris tenuis.

Nesidiocoris tenuis (Reuter) (Heteroptera: Miridae) is a tropical mirid bug used as a biocontrol agent in protected crops, including tomatoes. Although N. tenuis predates important insect pests, especially whitefly, it also causes damage by feeding on tomato plants when prey populations decline, resulting in significant economic losses for growers. The pest is now established in some all-year-round tomato crops in Europe and control measures involve the application of pesticides which are incompatible with current IPM programs. As part of future IPM strategies, the pheromone of N. tenuis was investigated. Volatile collections were made from groups and individuals of mated and unmated, females and males. In analyses of these collections by gas chromatography coupled with electroantennographic (EAG) recording from antennae of male bugs, two EAG-active components were detected and identified as 1-octanol and octyl hexanoate. Unlike other mirids, both male and female N. tenuis produced the two compounds, before and after mating, and both sexes gave EAG responses to both compounds. Furthermore, only octyl hexanoate was detected in whole body solvent washes from both sexes. These compounds are not related to the derivatives of 3-hydroxybutyrate esters found as pheromone components in other members of the Bryocrinae sub-family, and the latter could not be detected in volatiles from N. tenuis and did not elicit EAG responses. Nevertheless, experiments carried out in commercial glasshouses showed that traps baited with a blend of the synthetic pheromone components caught essentially only male N. tenuis, and significantly more than traps baited with octyl hexanoate alone. The latter caught significantly more N. tenuis than unbaited traps which generally caught very few bugs. Traps at plant height caught more N. tenuis males than traps 1 m above or at the base of the plants. The trap catches provided an indication of population levels of N. tenuis and were greatly reduced following an application of insecticide.

Effect of ß-1/3,1/6-glucan upon immune responses and bacteria in the gut of healthy common carp (Cyprinus carpio).

ß-glucans are frequently included in the diet of healthy common carp Cyprinus carpio as a pre-emptive measure for combatting disease. In order to study the effect this has on the relationship between the gut bacteria and host immune response, carp were maintained on either a ß-glucan free diet or feed containing 0.1% MacroGard®, a ß-1/3, 1/6-glucan, for up to 7 weeks and analysis of innate immune gene expression and molecular analysis of the gut bacteria was performed. The data reveals feeding of MacroGard® to healthy carp does not induce bactericidal innate immune gene expression in the gut but does appear to alter bacterial species richness that did not have a negative effect on overall health. Analysis of innate immune gene expression within the upper midgut revealed that there were significant changes over time in the expression of Interleukin (il)-1ß, inducible nitric oxide synthase (inos), mucin (muc2) and C-reactive protein (crp2). Diet did not affect the number of copies of the bacterial 16s rDNA gene in the gut, used as a as a measure of total bacteria population size. However, PCR-denaturing gradient gel electrophoresis (DGGE) analysis revealed a shift in bacterial species richness with MacroGard feeding. Bactericidal immune gene expression of crp2, muc2 and il-1ß was weakly correlated with gut bacteria population size indicating a potentially limited role of these genes in interacting with the gut bacteria in healthy carp in order to maintain gut homeostatic conditions. These findings highlight the importance of considering both host immunity and the microbiome together in order to fully elucidate the effeect of immunomodulants, such as ß-glucans, upon gut health.

Identification of Female Sex Pheromone for Monitoring the Barred Tooth Striped Moth, Trichopteryx polycommata, a Priority Conservation Species.

Pheromone-baited traps can be excellent tools for sensitive detection of insects of conservation concern. Here, identification of the sex pheromone of Trichopteryx polycommata (Denis & Schiffermüller, 1775), an under-recorded UK priority species, is reported. In analyses of extracts of the pheromone glands of female T. polycommata by gas chromatography coupled to electroantennographic recording from the antenna of a male moth, a single active component was detected. This was identified as (Z,Z)-6,9-nonadecadiene (Z,Z6,9-19:H) by comparison of its mass spectrum and retention times with those of the synthetic standard. In a pilot field trial in Kent, UK, T. polycommata males were caught in pheromone traps baited with lures loaded with 1 mg and 2 mg (Z,Z)-6,9-19:H. Optimum lure loading was identified in a further five trials in Kent, Sussex and Lancashire where lures of 0, 0.001, 0.01, 0.1, 1, 2, 5 and 10 mg loadings were tested. Traps baited with 1 to 10 mg of ZZ6,9-19:H caught significantly more T. polycommata than traps baited with 0 mg and 0.001 mg. In a pilot survey of T. polycommata using pheromone lures around Morecambe Bay, UK, T. polycommata males were captured at 122 new sites within the three counties where trials took place, demonstrating the potential of pheromone monitoring to increase knowledge of abundance, distribution and ecology of this elusive species.

Floral Odors and the Interaction between Pollinating Ceratopogonid Midges and Cacao.

Most plant species depend upon insect pollination services, including many cash and subsistence crops. Plants compete to attract those insects using visual cues and floral odor which pollinators associate with a reward. The cacao tree, Theobroma cacao, has a highly specialized floral morphology permitting pollination primarily by Ceratopogonid midges. However, these insects do not depend upon cacao flowers for their life cycle, and can use other sugar sources. To understand how floral cues mediate pollination in cacao we developed a method for rearing Ceratopogonidae through several complete lifecycles to provide material for bioassays. We carried out collection and analysis of cacao floral volatiles, and identified a bouquet made up exclusively of saturated and unsaturated, straight-chain hydrocarbons, which is unusual among floral odors. The most abundant components were tridecane, pentadecane, (Z)-7-pentadecene and (Z)-8-heptadecene with a heptadecadiene and heptadecatriene as minor components. We presented adult midges, Forcipomyia sp. (subgen. Forcipomyia), Culicoides paraensis and Dasyhelea borgmeieri, with natural and synthetic cacao flower odors in choice assays. Midges showed weak attraction to the complete natural floral odor in the assay, with no significant evidence of interspecific differences. This suggests that cacao floral volatiles play a role in pollinator behavior. Midges were not attracted to a synthetic blend of the above four major components of cacao flower odor, indicating that a more complete blend is required for attraction. Our findings indicate that cacao pollination is likely facilitated by the volatile blend released by flowers, and that the system involves a generalized odor response common to different species of Ceratopogonidae.

Evaluation of Host-Derived Volatiles for Trapping Culicoides Biting Midges (Diptera: Ceratopogonidae).

Culicoides biting midges (Diptera: Ceratopognidae) cause pain and distress through blood feeding, and transmit viruses that threaten both animal and human health worldwide. There are few effective tools for monitoring and control of biting midges, with semiochemical-based strategies offering the advantage of targeting host-seeking populations. In previous studies, we identified the host preference of multiple Culicoides species, including Culicoides impunctatus, as well as cattle-derived compounds that modulate the behavioral responses of C. nubeculosus under laboratory conditions. Here, we test the efficacy of these compounds, when released at different rates, in attracting C. impunctatus under field conditions in Southern Sweden. Traps releasing 1-octen-3-ol, decanal, phenol, 4-methylphenol or 3-propylphenol, when combined with carbon dioxide (CO2), captured significantly higher numbers of C. impunctatus compared to control traps baited with CO2 alone, with low release rates (0.1 mg h-1, 1 mg h-1) being generally more attractive. In contrast, traps releasing octanal or (E)-2-nonenal at 1 mg h-1 and 10 mg h-1 collected significantly lower numbers of C. impunctatus than control traps baited with CO2 only. Nonanal and 2-ethylhexanol did not affect the attraction of C. impunctatus when compared to CO2 alone at any of the release rates tested. The potential use of these semiochemicals as attractants and repellents for biting midge control is discussed.

Identification of Cattle-Derived Volatiles that Modulate the Behavioral Response of the Biting Midge Culicoides nubeculosus.

Identification of host-derived volatiles is an important step towards the development of novel surveillance and control tools for Culicoides biting midges. In this study, we identified compounds from headspace collections of cattle hair and urine that modulate the behavioral response of Culicoides nubeculosus, a research model species with a similar host-range as the vectors of Bluetongue disease and Schmallenberg disease in Europe. Combined gas chromatography and electroantennographic detection (GC-EAD) analysis revealed 23 bioactive compounds, of which 17, together with octanal, were evaluated in a two-choice behavioral assay in the presence of CO2. Decanal, 2-phenylethanal, 1-octen-3-ol, 2-ethylhexanol, 3-methylindole, phenol, and 3-ethylphenol elicited attraction of host seeking C. nubeculosus, whereas heptanal, octanal, nonanal, 3-propylphenol, and 4-propylphenol inhibited the insects' attraction to CO2, when compared to CO2 alone. 6-Methyl-5-hepten-2-one, 3-methylphenol, 4-methylphenol, and 4-ethylphenol elicited both attraction and inhibition. The behavioral responses were dependent on the concentration tested. Our results show that cattle-derived odors have the potential to be used for the manipulation of the behavior of Culicoides biting midges.

Methyl N,N-dimethylanthranilate and ethyl propionate: repellents effective against spotted wing drosophila, Drosophila suzukii.

BACKGROUND: Spotted wing drosophila, Drosophila suzukii (Matsumura) (Diptera: Drosophilidae), is an economically important pest of soft and stone fruit crops. The aim of this study was to identify repellents, formulated in dispensers, which could protect crops from D. suzukii. Fourteen potential repellents were screened against summer- and winter-morph D. suzukii through electroantennography and behavioural bioassays. Repellents effective in the laboratory were tested in polytunnels to determine their efficacy in reducing catches in fruit-baited traps. Further trials of three potential repellents were conducted to determine the distances over which repellent dispensers could reduce D. suzukii emergence in a strawberry crop. RESULTS: All 14 chemicals screened were detected by the antennae of both D. suzukii morphs. Hexyl acetate and geosmin both elicited a significantly greater corrected EAG response in summer morphs than winter morphs. Summer-morph D. suzukii were repelled by butyl acetate, ethyl propionate, methyl N,N-dimethyl anthranilate, geosmin, methyl salicylate, DEET and benzaldehyde at one or more doses test in laboratory bioassays. Winter morphs were repelled by ethyl propionate, methyl anthranilate, methyl N,N-dimethyl anthranilate, DEET, benzaldehyde and butyl anthranilate at one or more of the doses tested in the laboratory. Ethyl propionate, methyl N,N-dimethylanthranilate and benzaldehyde repelled both morphs from fruit-baited traps in polytunnel trapping trials. Ethyl propionate and methyl N,N-dimethylanthranilate reduced emergence of D. suzukii in a strawberry crop over 3-5 m. CONCLUSIONS: Ethyl propionate and methyl N,N-dimethylanthranilate may protect strawberry crops against D. suzukii. Future work should test these repellents in combination with attractants in a 'push-pull' strategy. © 2024 The Authors. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

Antagonistic and additive effect when combining biopesticides against the fall armyworm, Spodoptera frugiperda.

Fall armyworm, Spodoptera frugiperda (FAW) is a cosmopolitan crop pest species that has recently become established in sub-Saharan Africa and Southeast Asia. Current FAW control is almost entirely dependent on synthetic pesticides. Biopesticides offer a more sustainable alternative but have limitations. For example, pyrethrum is an effective botanical insecticide with low mammalian toxicity but is highly UV labile, resulting in a rapid loss of efficacy in the field. Beauveria bassiana is an entomopathogenic fungus that is more persistent, but there is a time lag of several days before it causes insect mortality and leads to effective control. The combination of these biopesticides could mitigate their drawbacks for FAW control. Here we evaluated the efficacy of pyrethrum and B. bassiana as individual treatments and in combination against 3rd instar FAW. Four different combinations of these two biopesticides were tested, resulting in an antagonistic relationship at the lowest concentrations of B. bassiana and pyrethrum (1 × 104 conidia mL-1 with 25 ppm) and an additive effect for the other 3 combined treatments (1 × 104 conidia mL-1 with 100 ppm and 1 × 105 conidia mL-1 with 25 ppm and 100 ppm pyrethrum). Additionally, a delay in efficacy from B. bassiana was observed when combined with pyrethrum as well as a general inhibition of growth on agar plates. These results appear to show that this particular combination of biopesticides is not universally beneficial or detrimental to pest control strategies and is dependent on the doses of each biopesticide applied. However, the additive effect shown here at specific concentrations does indicate that combining biopesticides could help overcome the challenges of persistence seen in botanical pesticides and the slow establishment of EPF, with the potential to improve effectiveness of biopesticides for IPM.

A morphologically distinct Phlebotomus argentipes population from active cutaneous leishmaniasis foci in central Sri Lanka.

Although the reported aetiological agent of cutaneous leishmaniasis (CL) in Sri Lanka is Leishmania donovani, the sandfly vector remains unknown. Ninety-five sandflies, 60 females and 35 males, collected in six localities in the district of Matale, central Sri Lanka, close to current active transmission foci of CL were examined for taxonomically relevant characteristics. Eleven diagnostic morphological characters for female sandflies were compared with measurements described for Indian and Sri Lankan sandflies, including the now recognised Phlebotomus argentipes sensu lato species complex. The mean morphometric measurements of collected female sandflies differed significantly from published values for P. argentipes morphospecies B, now re-identified as Phlebotomus annandalei from Delft Island and northern Sri Lanka, from recently re-identified P. argentipes s.s. sibling species and from Phlebotomus glaucus. Furthermore, analysis of underlying variation in the morphometric data through principal component analysis also illustrated differences between the population described herein and previously recognised members of the P. argentipes species complex. Collectively, these results suggest that a morphologically distinct population, perhaps most closely related to P. glaucus of the P. argentipess. I. species complex, exists in areas of active CL transmission. Thus, research is required to determine the ability of this population of flies to transmit cutaneous leishmaniasis.

Live Drosophila melanogaster Larvae Deter Oviposition by Drosophila suzukii.

The worldwide invasive insect pest, Drosophila suzukii Matsumura (spotted-wing Drosophila), lays eggs in soft and stone fruit before harvest. Hatched larvae cause fruit collapse and significant economic losses. Current control methods rely primarily on foliar insecticide applications, which are not sustainable long-term solutions due to regulatory restrictions and the risk of insecticide resistance developing. We showed before that D. suzukii were deterred from laying eggs on artificial media previously visited by its sister species-Drosophila melanogaster. In the current study, laboratory choice test experiments were conducted to identify which D. melanogaster life stage (eggs, larvae, or adult) deterred D. suzukii oviposition. We demonstrated that the presence of live D. melanogaster larvae on the egg-laying media consistently deterred D. suzukii oviposition. Drosophila melanogaster cuticular hydrocarbons (CHCs) were examined as candidate for the oviposition deterrent. CHCs of larval and adult D. melanogaster and D. suzukii were analyzed. In both species, the composition of the CHCs of larvae was similar to that of adults, although quantities present were much lower. Furthermore, the CHC profiles of the two species were markedly different. However, when assayed as deterrents in the laboratory choice test experiment, CHC extracts from D. melanogaster did not deter oviposition by D. suzukii.

Insecticide-impregnated netting: A surface treatment for killing Lutzomyia longipalpis (Diptera: Psychodidae), the vector of Leishmania infantum.

The sand fly Lutzomyia longipalpis is the main vector of Leishmania infantum in Brazil. Synthetic male-produced sex/aggregation pheromone co-located with micro-encapsulated λ-cyhalothrin in chicken sheds can significantly reduce canine infection and sand fly densities in a lure-and-kill strategy. In this study, we determined if insecticide-impregnated netting (IN) could replace insecticide residual spraying (IRS). We compared numbers of Lu. longipalpis attracted and killed in experimental and real chicken sheds baited with pheromone and treated with a 1 m2 area of either insecticide spray or netting. First, we compared both treatments in experimental sheds to control mortality established from light trap captures. We then compared the long-term killing effect of insecticide spray and netting, without renewal, in experimental sheds over a period of 16 weeks. Finally, a longitudinal intervention study in real chicken sheds compared the numbers and proportions of Lu. longipalpis collected and killed before and after application of both treatments. In Experiment 1, a higher proportion of males and females captured in IRS- and IN-treated sheds were dead at 24 h compared to controls (P < 0.05). No difference was found in the proportion of females killed in sheds treated with IN or IRS (P = 0.15). A slightly higher proportion of males were killed by IRS (100%) compared to IN (98.6%; P < 0.05). In Experiment 2, IN- and IRS-treated traps were equally effective at killing females (P = 0.21) and males (P = 0.08). However, IRS killed a significantly higher proportion of females and males after 8 (P < 0.05) and 16 (P < 0.05) weeks. In Experiment 3, there was no significant difference between treatments in the proportion of females killed before (P = 0.88) or after (P = 0.29) or males killed before (P = 0.76) or after (P = 0.73) intervention. Overall, initially the IN was as effective as IRS at killing female and male Lu. longipalpis in both experimental and real chicken sheds. However, the relative lethal effect of the IN deteriorated over time when stored under prevailing environmental conditions.

Influence of light and kairomone baiting systems on trap collections of biting midges in southern Sweden.

Effective surveillance is essential for protecting livestock from Culicoides biting midges and the viruses they transmit. The objective of this study was to determine how the baiting system used in traps (UV, incandescent light, incandescent light with CO2 , and incandescent light with CO2 and 1-octen-3-ol) influences estimates of midge population abundance, parity, and diel activity. This was achieved through a standardized trapping protocol conducted in three habitats in Sweden. UV light traps caught the most Culicoides species and more C. obsoletus complex females than incandescent light traps. Traps baited with CO2 plus 1-octen-3-ol caught more female C. impunctatus than incandescent light traps. No consistent effect of bait type was found on C. obsoletus parity rate, as estimated from the proportion of midges with presence or absence of pigmentation. Midge activity, as reflected by trap catches, peaked between -3 h and +3 h relative to sunset, with UV traps catching significantly more female C. obsoletus complex and C. impunctatus at and after sunset than before sunset. We conclude that baiting system can influence biting midge collections, even using identical traps. Effective surveillance may require more than one bait type and kairomones to attract species that do not feed exclusively on cattle.

Additive Effect of Botanical Insecticide and Entomopathogenic Fungi on Pest Mortality and the Behavioral Response of Its Natural Enemy.

Sustainable agricultural intensification employs alternatives to synthetic insecticides for pest management, but these are not always a direct replacement. Botanical insecticides, for example, have rapid knockdown but are highly labile and while biological pesticides are more persistent, they are slow acting. To mitigate these shortcomings, we combined the entomopathogenic fungus (EPF) Metarhizium anisopliae with pyrethrum and evaluated their efficacy against the bean aphid, Aphis fabae. To ascertain higher trophic effects, we presented these treatments to the parasitoid, Aphidius colemani, on an aphid infested plant in a Y-tube olfactometer and measured their preferences. Aphid mortality was significantly higher than controls when exposed to EPF or pyrethrum but was greater still when exposed to a combination of both treatments, indicating an additive effect. This highlights the potential for applications of pyrethrum at lower doses, or the use of less refined products with lower production costs to achieve control. While parasitoids were deterred by aphid infested plants treated with EPF, no preference was observed with the combination pesticide, which provides insight into the importance that both application technique and timing may play in the success of this new technology. These results indicate the potential for biorational pesticides that combine botanicals with EPF.

Sand fly synthetic sex-aggregation pheromone co-located with insecticide reduces the incidence of infection in the canine reservoir of visceral leishmaniasis: A stratified cluster randomised trial.

OBJECTIVE: To evaluate the efficacy of a synthetic sex-aggregation pheromone of the sand fly vector Lu. longipalpis, co-located with residual insecticide, to reduce the infection incidence of Leishmania infantum in the canine reservoir. METHODS: A stratified cluster randomised trial was designed to detect a 50% reduction in canine incident infection after 24 months in 42 recruited clusters, randomly assigned to one of three intervention arms (14 cluster each): synthetic pheromone + insecticide, insecticide-impregnated dog collars, or placebo control. Infection incidence was measured by seroconversion to anti-Leishmania serum antibody, Leishmania parasite detection and canine tissue parasite loads. Changes in relative Lu. longipalpis abundance within households were measured by setting three CDC light traps per household. RESULTS: A total 1,454 seronegative dogs were followed-up for a median 15.2 (95% C.I.s: 14.6, 16.2) months per cluster. The pheromone + insecticide intervention provided 13% (95% C.I. 0%, 44.0%) protection against anti-Leishmania antibody seroconversion, 52% (95% C.I. 6.2%, 74·9%) against parasite infection, reduced tissue parasite loads by 53% (95% C.I. 5.4%, 76.7%), and reduced household female sand fly abundance by 49% (95% C.I. 8.2%, 71.3%). Variation in the efficacy against seroconversion varied between trial strata. Equivalent protection attributed to the impregnated-collars were 36% (95% C.I. 14.4%, 51.8%), 23% (95% C.I. 0%, 57·5%), 48% (95% C.I. 0%, 73.4%) and 43% (95% C.I. 0%, 67.9%), respectively. Comparison of the two interventions showed no statistically consistent differences in their efficacies; however, the errors were broad for all outcomes. Reductions in sand fly numbers were predominant where insecticide was located (chicken and dog sleeping sites), with no evidence of insecticide-induced repellence onto humans or dogs. CONCLUSION: The synthetic pheromone co-located with insecticide provides protection particularly against canine L. infantum parasite transmission and sand fly vector abundance. The effect estimates are not dissimilar to those of the insecticide-impregnated collars, which are documented to reduce canine infection incidence, human infection and clinical VL disease incidence, in different global regions. The trialled novel lure-and-kill approach is a low-cost potential vector control tool against ZVL in the Americas.

Multi-modal analysis of courtship behaviour in the old world leishmaniasis vector Phlebotomus argentipes.

BACKGROUND: The sand fly Phlebotomus argentipes is arguably the most important vector of leishmaniasis worldwide. As there is no vaccine against the parasites that cause leishmaniasis, disease prevention focuses on control of the insect vector. Understanding reproductive behaviour will be essential to controlling populations of P. argentipes, and developing new strategies for reducing leishmaniasis transmission. Through statistical analysis of male-female interactions, this study provides a detailed description of P. argentipes courtship, and behaviours critical to mating success are highlighted. The potential for a role of cuticular hydrocarbons in P. argentipes courtship is also investigated, by comparing chemicals extracted from the surface of male and female flies. PRINCIPAL FINDINGS: P. argentipes courtship shared many similarities with that of both Phlebotomus papatasi and the New World leishmaniasis vector Lutzomyia longipalpis. Male wing-flapping while approaching the female during courtship predicted mating success, and touching between males and females was a common and frequent occurrence. Both sexes were able to reject a potential partner. Significant differences were found in the profile of chemicals extracted from the surface of males and females. Results of GC analysis indicate that female extracts contained a number of peaks with relatively short retention times not present in males. Extracts from males had higher peaks for chemicals with relatively long retention times. CONCLUSIONS: The importance of male approach flapping suggests that production of audio signals through wing beating, or dispersal of sex pheromones, are important to mating in this species. Frequent touching as a means of communication, and the differences in the chemical profiles extracted from males and females, may also indicate a role for cuticular hydrocarbons in P. argentipes courtship. Comparing characteristics of successful and unsuccessful mates could aid in identifying the modality of signals involved in P. argentipes courtship, and their potential for use in developing new strategies for vector control.

Synthetic sex pheromone in a long-lasting lure attracts the visceral leishmaniasis vector, Lutzomyia longipalpis, for up to 12 weeks in Brazil.

Current control methodologies have not prevented the spread of visceral leishmaniasis (VL) across Brazil. Here, we describe the development of a new tool for controlling the sand fly vector of the disease: a long-lasting lure, which releases a synthetic male sex pheromone, attractive to both sexes of Lutzomyia longipalpis. This device could be used to improve the effectiveness of residual insecticide spraying as a means of sand fly control, attracting L. longipalpis to insecticide-treated animal houses, where they could be killed in potentially large numbers over a number of weeks. Different lure designs releasing the synthetic pheromone (±)-9-methylgermacrene-B (CAS 183158-38-5) were field-tested in Araçatuba, São Paulo (SP). Experiments compared numbers of sand flies caught overnight in experimental chicken sheds with pheromone lures, to numbers caught in control sheds without pheromone. Prototype lures, designed to last one night, were first used to confirm the attractiveness of the pheromone in SP, and shown to attract significantly more flies to test sheds than controls. Longer-lasting lures were tested when new, and at fortnightly intervals. Lures loaded with 1 mg of pheromone did not attract sand flies for more than two weeks. However, lures loaded with 10 mg of pheromone, with a releasing surface of 15 cm2 or 7.5 cm2, attracted female L. longipalpis for up to ten weeks, and males for up to twelve weeks. Approximately five times more sand flies were caught with 7.5 cm2 10 mg lures when first used than occurred naturally in non-experimental chicken resting sites. These results demonstrate that these lures are suitably long-lasting and attractive for use in sand fly control programmes in SP. To our knowledge, this is the first sex pheromone-based technology targeting an insect vector of a neglected human disease. Further studies should explore the general applicability of this approach for combating other insect-borne diseases.

Insecticide-impregnated netting as a potential tool for long-lasting control of the leishmaniasis vector Lutzomyia longipalpis in animal shelters.

BACKGROUND: Leishmaniasis remains a serious neglected disease, with more than 350 million people potentially at risk worldwide. Control strategies often rely on spraying residual insecticides to target populations of the sand fly vectors that transmit Leishmania parasites when blood-feeding. These programmes are often difficult to sustain effectively, as sand fly resting sites must be resprayed on a regular basis. Here, we investigate whether application of insecticide-impregnated netting to a surface could act as an alternative to residual spraying for controlling the American visceral leishmaniasis vector Lutzomyia longipalpis. METHODS: Female L. longipalpis from our laboratory colony were exposed for 1 h to three treatments applied to plywood surfaces: 2% permethrin-impregnated netting (Olyset®), 20 mg a.i.m-2 micro-encapsulated lambda-cyhalothrin (Demand CS®) and a no-treatment control. We compared the speed at which these treatments acted, by measuring the percentage of sand flies killed both immediately after exposure to the treatment for 1 hour, as well as the number that had died 24 h after the 1 hour exposure. We repeated the experiment at 6 and 12 months following application to test the effectiveness of each treatment over time. RESULTS: When first applied, the lambda-cyhalothrin killed more sand flies in the first hour than the permethrin-impregnated netting. However, the effectiveness of the lambda-cyhalothrin diminished over time, so that there was no difference between the two treatments at 12 months. Both killed more sand flies than the control. When measured 24 h following exposure, both test treatments had killed close to 100% of sand flies when first applied, but while the lethal effect of the netting was maintained at close to 100% over 12 months, the effectiveness of the residual insecticide diminished to approximately 80% after 6 months. CONCLUSIONS: The results of these initial laboratory experiments indicate that covering surfaces with insecticide impregnated netting material may provide a longer-lasting solution for killing sand flies than residual spraying. Field trials are needed to identify the feasibility of treating surfaces with netting or similar impregnated materials as part of a control program. In targeting L. longipalpis, the greatest benefits may be seen in treating animal sheds with netting, where these sand flies aggregate in large numbers, and which can be difficult to treat repeatedly by conventional spraying.

Attraction of the cutaneous leishmaniasis vector Nyssomyia neivai (Diptera: Psychodidae) to host odour components in a wind tunnel.

BACKGROUND: Laboratory studies of host-seeking olfactory behaviour in sandflies have largely been restricted to the American visceral leishmaniasis vector Lutzomyia longipalpis. In comparison, almost nothing is known about the chemical ecology of related species, which transmit American cutaneous leishmaniasis (ACL), due in part to difficulties in raising these insects in the laboratory. Understanding how ACL vectors locate their hosts will be essential to developing new vector control strategies to combat this debilitating disease. METHODS: This study examined host-odour seeking behaviour of the ACL vector Nyssomyia neivai (Pinto) (=Lutzomyia neivai) using a wind tunnel olfactometer. The primary aim was to determine whether field-collected female N. neivai would respond to host odours in the laboratory, thereby eliminating the need to maintain colonies of these insects for behavioural experiments. Responses to two key host odour components, 1-octen-3-ol and lactic acid, and a commercially-available mosquito lure (BG-Lure™) were assessed and compared relative to an air control. We also tested whether trials could be conducted outside of the normal evening activity period of N. neivai without impacting on fly behaviour, and whether the same flies could be used to assess baseline responses to air without affecting responses to octenol, thereby reducing the number of flies required for experiments. RESULTS: Octenol was found to both activate host-seeking behaviour and attract female N. neivai in the wind tunnel, while lactic acid elicited weaker responses of activation and attractiveness under identical conditions. The BG-Lure did not activate or attract N. neivai under test conditions. Further experiments showed that sandfly behaviour in the wind tunnel was not affected by time of day, such that experiments need not be restricted to nocturnal hours. Moreover, using the same flies to measure both baseline responses to air and attraction to test compounds did not affect odour-seeking behaviour. CONCLUSIONS: The results of this study demonstrate that N. neivai taken from the field are suitable for use in laboratory olfactometer experiments. It is hoped this work will facilitate further research into chemical ecology of this species, and other ACL vectors.