Evaluation of several novel alkynols, alkenols, and selected host odor blends as attractants to female Aedes albopictus and Culex quinquefasciatus.

The compound 1-octen-3-ol is a strong attractant for some mosquito species. Based on chemical structure, this may be due to a terminal site of unsaturation or high electron density, a structural capability for hydrogen bonding, e.g., -OH, -NH2, NHR, NR2, etc., a saturated hydrocarbon chain of a certain minimum length, and a certain relative distance between the region of high electron density and the alcohol (or other hydrogen-bonding) functional group. Based on this hypothesis, 4 novel alkynol (triple-bonded) analogs were synthesized and evaluated alone or in combination with acetone and dimethyl disulfide, and with and without carbon dioxide in Mosquito Magnet-X suction traps. Attraction of laboratory-reared adult Aedes albopictus and Culex quinquefasciatus to these analogs and combinations was compared to 1-octen-3-ol as a standard in semi-field trials. For both species none of the alkynols, with and without carbon dioxide or acetone and dimethyl disulfide, were significantly different from 1-octen-3-ol. The compounds 2-octyn-4-ol and 2-nonyn-4-ol alone and with carbon dioxide suppressed Cx. quinquefasciatus collections. An additional 6 alkenol (double-bonded) analogs were tested in mixtures with 3-n-propylphenol and 4-methylphenol in a ratio of 4:1:8, respectively. Using the same trapping methods, Cx. quinquefasciatus catches containing 3-decen-1-ol were increased nearly 3-fold when combined with carbon dioxide. Aedes albopictus collections in traps with the 3-decen-1-ol/phenol mixture and carbon dioxide were significantly greater than similar traps with 1-octen-3-ol. Traps baited with the phenol blends that incorporated (Z)-3-nonen-1-ol, (Z)-8-nonen-3-ol, or 1-octen-3-ol were considerably suppressed in the presence of carbon dioxide.

Relationship between mosquito (Diptera: Culicidae) landing rates on a human subject and numbers captured using CO2-baited light traps.

Capture rates of insectary-reared female Aedes albopictus (Skuse), Anopheles quadrimaculatus Say, Culex nigripalpus Theobald, Culex quinquefasciatus Say and Aedes triseriatus (Say) in CDC-type light traps (LT) supplemented with CO2 and using the human landing (HL) collection method were observed in matched-pair experiments in outdoor screened enclosures. Mosquito responses were compared on a catch-per-unit-effort basis using regression analysis with LT and HL as the dependent and independent variables, respectively. The average number of mosquitoes captured in 1 min by LT over a 24-h period was significantly related to the average number captured in 1 min by HL only for Cx. nigripalpus and Cx. quinquefasciatus. Patterns of diel activity indicated by a comparison of the mean response to LT and HL at eight different times in a 24-h period were not superposable for any species. The capture rate efficiency of LT when compared with HL was ≤15% for all mosquitoes except Cx. quinquefasciatus (43%). Statistical models of the relationship between mosquito responses to each collection method indicate that, except for Ae. albopictus, LT and HL capture rates are significantly related only during certain times of the diel period. Estimates of mosquito activity based on observations made between sunset and sunrise were most precise in this regard for An. quadrimaculatus and Cx. nigripalpus, as were those between sunrise and sunset for Cx. quinquefasciatus and Ae. triseriatus.

Efficacy of commercial mosquito traps in capturing phlebotomine sand flies (Diptera: Psychodidae) in Egypt.

Four types of commercial mosquito control traps, the Mosquito Magnet Pro (MMP), the Sentinel 360 (S360), the BG-Sentinel (BGS), and the Mega-Catch Ultra (MCU), were compared with a standard Centers for Disease Control and Prevention (CDC) light trap for efficacy in collecting phlebotomine sand flies (Diptera: Psychodidae) in a small farming village in the Nile River Valley 10 km north of Aswan, Egypt. Each trap was baited with either carbon dioxide (CO2) from combustion of butane gas (MMP), dry ice (CDC and BGS traps), light (MCU and S360), or dry ice and light (CDC). Traps were rotated through five sites in a5 x 5 Latin square design, repeated four times during the height of the sand fly season (June, August, and September 2007) at a site where 94% of sand flies in past collections were Phlebotomus papatasi (Scopoli). A total of 6,440 sand flies was collected, of which 6,037 (93.7%) were P. papatasi. Of the CO2-baited traps, the BGS trap collected twice as many P. papatasi as the MMP and CDC light traps, and at least three times more P. papatasi than the light-only MCU and S360 traps (P < 0.05). Mean numbers (+/- SE) of P. papatasi captured per trap night were as follows: BGS 142.1 (+/- 45.8) > MMP 56.8 (+/- 9.0) > CDC 52.3 (+/- 6.1) > MCU 38.2 (+/- 6.4) > S360 12.6 (+/- 1.8). Results indicate that several types of commercial traps are suitable substitutes for the CDC light trap in sand fly surveillance programs.

Residual efficacy of field-applied permethrin, d-phenothrin, and resmethrin on plant foliage against adult mosquitoes.

Backpack sprayer applications of permethrin, d-phenothrin, and resmethrin to vegetation and plants at Anastasia Island, St. Augustine, FL, were evaluated for duration of residual efficacy against adult mosquitoes. All treatments produced 100% mortality (24 h) of mosquitoes in test cages placed within the vegetation. At 48 h and 1 wk posttreatment, insecticide treatments resulted in 70-100% reduction of adult mosquitoes caught by Centers for Disease Control and Prevention traps baited with 1-octen-3-ol. Insecticide residues in excised leaves from both treated and control areas of the study sites were evaluated against adult female Culex quinquefasciatus by laboratory bioassay. Permethrin produced 90% mortality up to 1 wk postapplication. Both d-phenothrin and resmethrin produced nearly this level of mortality for a much shorter duration of <48 h postapplication, with residual effects dropping significantly thereafter. Average insecticide concentrations in leaves were quantified by gas chromatography/mass spectroscopy, and some correlation was observed between chemical and biological results.

Evaluation of the enantiomers of 1-octen-3-ol and 1-octyn-3-ol as attractants for mosquitoes associated with a freshwater swamp in Florida, U.S.A.

Field studies were conducted at wooded wetlands in Gainesville, FL, U.S.A., to assess responses of natural populations of adult mosquitoes (Diptera: Culicidae) to American Biophysics MM-X and Coleman MD-2500 traps baited with enantiomers of 1-octen-3-ol, a naturally occurring compound, and 1-octyn-3-ol, a closely related synthetic compound. Overall, the same species of mosquitoes were attracted by all enantiomers, although the (R)-(+) isomer of octenol generally attracted more species, and it is the isomer produced in greatest proportion in nature. Traps baited with the R-enantiomer caught greater numbers of mosquitoes than those baited with the S-enantiomer of each compound, whereas traps baited with S-enantiomers were equally or slightly less attractive than those baited with carbon dioxide only.

Attraction of mosquitoes to volatiles associated with blood.

Responses of the mosquitoes Aedes aegypti, Culex quinquefasciatus, and Culex nigripalpus to volatiles and compounds associated with bovine and avian blood that were presented in collagen membranes were evaluated in olfactometer and landing assays. The presence of attractants produced by blood was supported by more attraction of all species to blood than water controls in the olfactometer. Females of Ae. aegypti and Cx. quinquefasciatus were more attracted to bovine blood than to avian blood, but there was no difference in Cx. nigripalpus responses. In landing assays, significantly more females of all species landed on casings with blood than on water controls. There was no difference in landing of Ae. aegypti on bovine or avian blood. However, significantly more females of Cx. quinquefasciatus and Cx. nigripalpus landed on avian blood compared to bovine blood. Blood presented in collagen casings was an effective method for evaluating in-flight attraction and landing in all three species. In the olfactometer, several individual compounds elicited attraction in all species, but none were as attractive as blood for all species. In landing assays, several organic acids and sulfides elicited landing, with Ae. aegypti responding to the greatest number of compounds. These assay methods are effective for evaluation of volatile compounds from blood, and although responses were obtained to several compounds, none were as effective as blood in the olfactometer and landing assays.

Laboratory evaluation of avian odors for mosquito (Diptera: Culicidae) attraction.

Attraction of Culex quinquefasciatus Say, Culex tarsalis Coquillett, Culex nigripalpus Theobald, and Aedes aegypti (L.) to avian and other host odors was investigated in a dual-port olfactometer. Although attraction to a human arm was high for Ae. aegypti (>80%) and low for all Culex spp. (<25%), all species responded similarly to a chicken (55.3-73.6%). Responses of Ae. aegypti, Cx. quinquefasciatus, and Cx. nigripalpus to feathers were low (<20%) but greater than to controls. There was no difference in attraction of Cx. tarsalis to feathers or controls. Responses to CO2 (5 ml/min) were low for all species (<15%) except Cx. tarsalis, which were moderate (24.5%). When feathers were combined with CO2, the resulting attraction was additive or lower than responses to feather and CO2 alone for all species except for Cx. tarsalis, which had responses that were three-fold greater than expected if responses were additive. The CO2-feather treatments were less attractive than a chicken for all species. When olfactometer assays were extended from 3 to 20 min, responses by Ae. aegypti significantly increased to a chicken and CO2 and attraction of Cx. quinquefasciatus significantly greater to chickens, CO2, and feathers. None of the volatile compounds previously identified from feathers or uropygial glands tested were attractive. Both feather-rubbed cotton balls and hexane extracts of feathers were attractive and as attractive as feathers; however, ether extracts were not attractive. Feathers clearly contribute to the attraction of host-seeking Culex spp., and future studies will focus on identification of the attractant compounds.

Sex pheromone of the tsetse species, Glossina austeni: isolation and identification of natural hydrocarbons, and bioassay of synthesized compounds.

Copulatory responses of male Glossina austeni (Newstead) (Diptera: Glossinidae), that were elicited after contact with frozen female tsetse, were not observed after solvent washing of cuticular lipids. Chromatographic analysis of extracts from laboratory-reared and field-collected G. austeni females yielded natural hydrocarbons that were highly stimulatory to males. Most of this activity was produced by compounds in the alkene fraction. Gas chromatograms (GC) contained five natural alkenes; these were separated by preparative GC for bioassays conducted in Tanzania. The two major alkenes were identified using gas chromatography-mass spectrometry (GC-MS) to be 13,17-dimethyltritriacont-1-ene and 13,17-dimethylpentatriacont-1-ene, after the samples had undergone derivatization using dimethyl disulphide and saturation with deuterium. These alkenes and natural alkanes were quantified from G. austeni of both sexes from laboratory and field samples to confirm that their presence was consistent in this species. Trials of synthetic samples resulted in the order of biological activity for the stereoisomers of 13,17-dimethyltritriacont-1-ene as follows: S,R-33:1 > R,S- 33:1 > S,S-33:1 > R,R-33:1. Dose-response data showed an ED(50) at 5 microg per treated, solvent-washed male decoy. Of the four stereoisomers of 13,17-dimethylpentatriacont-1-ene, R,R-35:1 showed the most activity. This is the first report of alkene-induced sexual activity in males of the genus Glossina.

Distinctive hydrocarbons of the black dump fly, Hydrotaea aenescens (Diptera: Muscidae).

Hydrotaea aenescens (Wiedemann), the black dump fly, is a potential biological control agent originally from the western hemisphere, now found in many parts of the Palearctic region except for the United Kingdom, where it cannot be imported for any reason. A complication of classical biological control is the problem of strain identification, as one must be able to somehow mark or follow a particular strain that has been introduced into the field or is contemplated for release. Gas chromatographic analysis of the surface hydrocarbons of pooled and individual dump fly adults resulted in reproducible hydrocarbon patterns that differentiated widely distributed strains of H. aenescens and showed similarities between strains that were related. Sexual dimorphism was observed in the surface hydrocarbons. Conspecific similarities included identities of the hydrocarbons found in colony material collected worldwide, with differences being found in the quantities of compounds present.

Analysis of human skin emanations by gas chromatography/mass spectrometry. 2. Identification of volatile compounds that are candidate attractants for the yellow fever mosquito (Aedes aegypti).

Volatile compounds emanated from human skin were studied by gas chromatography/mass spectrometry (GC/MS). The purpose of this study was to identify compounds that may be human-produced kairomones which are used for host location by the mosquito, Aedes aegypti (L.). The procedure used to collect volatiles was chosen because of prior knowledge that attractive substances can be transferred from skin to glass by handling. Laboratory bioassays have shown that the residuum on the glass remains attractive to mosquitoes until the compounds of importance evaporate. The sampling and analytical procedures modeled the above-cited process as closely as possible except that the evaporation of compounds from the glass surface was accomplished by thermal desorption from glass beads in a heated GC injection port. This made possible the solventless injection of volatiles onto the column. The compounds were cryofocused on the head of the column with liquid nitrogen prior to GC separation. A single stage of mass spectrometry on a triple quadrupole instrument was used for mass analysis. A combination of electron ionization and pulsed positive ion/negative ion chemical ionization modes on two different GC columns (one polar, one relatively nonpolar) was used to identify most of the 346 compound peaks detected by this technique.

Analysis of human skin emanations by gas chromatography/mass spectrometry. 1. Thermal desorption of attractants for the yellow fever mosquito (Aedes aegypti) from handled glass beads.

Handled glass has the ability to collect and concentrate nonaqueous human skin emanations while minimizing the collection of aqueous perspiration. Compounds originating from the skin and collected on glass have previously been found to attract the Aedes aegypti species of mosquito. Therefore, glass beads were used as the medium to collect skin emanations from humans for subsequent chemical analysis. This process consisted of a 5-15-min collection of sample on glass beads, followed by loading the beads into a gas chromatograph (GC) injector insert for subsequent desorption of the collected compounds onto the GC column. After cryofocusing by liquid nitrogen at the head of the column, the thermally desorbed compounds were analyzed by GC/MS. Microscale purge and trap introduction was also used to provide complementary information. In this case, the beads are held in a round-bottom flask, purged with nitrogen, and heated as the concentrator collects the headspace above the beads. The chromatograms produced by both of these sample introduction methods demonstrate good resolution of a complex sample. Cryofocusing volatiles from handled glass allowed identification of lactic acid, aliphatic fatty acids, and other polar to nonpolar compounds of moderate volatility while purge and trap allowed detection of nonpolar to moderately polar compounds of high volatility.

Gas chromatography/mass spectrometry analysis of the cuticular hydrocarbons from parasitic wasps of the genus Muscidifurax.

Parasitic Hymenoptera can be difficult to identify by conventional taxonomic techniques. Examination of the cuticular hydrocarbons (CHCs) provides a basis for chemotaxonomic differentiation, which may lead to the discovery of pheromones, and can be a means of examining colonies for species cross-contamination. The parasitic wasps examined were Muscidifurax raptor, M. zaraptor, M. uniraptor, and the gregarious form of M. raptorellus. Species within the genus Muscidifurax, as well as the sex, can clearly be differentiated by examining the gas chromatograms of the CHCs. Identification of the alkanes by mass spectrometry shows uncommon dimethylalkanes and trimethylalkanes for members of the genus. The methyl branched cuticular hydrocarbons of these insects are rare compared to those found on insects reported in the literature, but are present in significant amounts on these insects. Additionally, sexual dimorphism is observed in long chain alkanes (C21-C39) present on male and female cuticular surfaces for these species. Females tend to have cuticular hydrocarbons with methyl branches located externally on the carbon backbone chain for dimethyl-, trimethyl-, and tetramethylalkanes, whereas males tend to have dimethyl- and trimethylalkanes located internally on the hydrocarbon backbone chains. Mass spectra of novel and rare methyl branched compounds identified on these parasitoids are presented.

Analysis of the cuticular hydrocarbons among species of the Anopheles quadrimaculatus complex (Diptera: Culicidae).

Cuticular hydrocarbons were extracted from females of 5 species of the Anopheles quadrimaculatus complex and studied by gas chromatography and mass spectrometry. The data were analyzed by multivariate techniques to determine the degree of divergence in hydrocarbon patterns and to develop models that allow the discrimination of these species. Anopheles quadrimaculatus Say, Anopheles smaragdinus Reinert, and Anopheles maverlius Reinert could be separated at 100% from each other and from Anopheles diluvialis Reinert and Anopheles inundatus Reinert; however, separation of An. diluvialis from An. inundatus was 80% using a 2-way model.