Monitoring temporal fluctuations of Culex quinquefasciatus using oviposition traps containing attractant and larvicide in an urban environment in Recife, Brazil

The use of attractants and larvicides in oviposition traps is of practical interest for the surveillance and control of urban mosquitoes. In addition to increasing the safety of the traps, this combination is essential for an attract-and-kill control strategy based on trapping mosquito eggs. The combination of Bacillus thuringiensis var. israelensis (Bti) and grass infusion (GI) vs. GI alone were tested for their ability to attract in paired BR-OVT traps in the backyards of 10 houses in Recife, Brazil, for a period of 45 days. Results show that females prefer to oviposit in traps containing Bti (363 compared with 251 egg rafts over 45 days). Results from a one-year trial on the efficacy of BR-OVT traps loaded with GI and Bti as a sampling tool to monitor temporal fluctuations in the population densities of Culex quinquefasciatus in an urban environment are also reported. From December 2006-January 2007, one trap per home was installed and maintained for 348 consecutive days in 134-151 houses located in three urban blocks. Throughout the one-year field trial a total of 43,151 Culex egg rafts were collected in the traps. The data show that BR-OVT loaded with GI and Bti is sensitive enough to demonstrate continuous reproductive activity of Cux. quinquefasciatus in the study area throughout the year and to monitor temporal fluctuations in population density.     

Comparative responses of ovipositing Anopheles gambiae and Culex quinquefasciatus females to the presence of Culex egg rafts and larvae

Field observations have demonstrated that gravid Anopheles gambiae Giles s.s. (Diptera: Culicidae) are selective in their choice of oviposition sites. For example, immature stages of An. gambiae s.s. are rarely found in water that contains Culex quinquefasciatus Say immatures. The possibility that this may, in part at least, reflect a response by ovipositing An. gambiae s.s. females to volatile signals associated with Culex juveniles was evaluated by testing the response of An. gambiae s.s. females to varying densities of Cx. quinquefasciatus egg rafts and/or larvae in oviposition choice assays. For comparison, the oviposition choices of Cx. quinquefasciatus to conspecific egg rafts and/or larvae were similarly assayed. At a low density of Cx. quinquefasciatus egg rafts (1-15 egg rafts/100 mL water), An. gambiae s.s. females laid more eggs in the treatment water than in the control, with a maximum of twice as many in the treatment water at 5 egg rafts/100 mL water. At higher egg raft densities and in all treatments that included Cx. quinquefasciatus larvae, oviposition decreased significantly in the treatment dishes in a density-dependent manner. As previous studies have indicated, ovipositing Cx. quinquefasciatus females were attracted to and laid egg rafts in dishes containing conspecific egg rafts and, interestingly, also in dishes containing larvae.     

Odour-mediated upwind flight of Culex quinquefasciatus mosquitoes elicited by a synthetic attractant

Abstract A wind tunnel and video equipment are used to study the long-range and close-range responses of gravid females of Culex quinquefasciatus Say to a synthetic pheromone, eryrthro -6-acetoxy-5-hexadecanolide. In response to water and acetoxyhexadecanolide, together or independently, in the presence of wind, females follow meandering flight paths upwind. Females have a higher rate of turning and a lower flight-speed when landing at a site containing pheromone than at a comparable site without pheromone. Females stay longer at oviposition sites containing the attractant than at sites with no attractant.     

Non-larvicidal effects of Bacillus thuringiensis israelensis and Bacillus sphaericus on oviposition and adult mortality of Culex quinquefasciatus Say (Diptera: Culicidae).

Two microbial mosquito larvicides, Bacillus thuringiensis ssp. israelensis (Bti) and Bacillus sphaericus (Bsph), have been shown to be highly effective in controlling mosquito larvae and have been used in larvicidal programs for many years. In exploring other modes of action of these agents, we studied the ovipositional response of Bsph susceptible and resistant Culex quinquefasciatus to aqueous suspensions of Bti and Bsph water dispersible granules (WDG). We quantified the level of mortality of adult mosquitoes caused by exposure to Bti and Bsph suspensions during oviposition. Significantly lower numbers of egg rafts were laid and collected from the treatments than the control regimen. There was an inverse relationship between Bacillus product concentrations and oviposition. As the concentration of Bti or Bsph increased from 0.0 to 2.0 mg/L, treated waters received progressively fewer egg rafts. In addition to the negative effects of Bacillus on oviposition, both male and female adult mosquitoes suffered high mortality on landing and imbibing on Bti and Bsph suspensions, the extent of mortality directly proportional to concentration. These two microbial agents used solely as mosquito larvicides thus have the additional benefits of reducing mosquito oviposition and killing adult mosquitoes, especially gravid females that come in contact with the treated water either for oviposition or drinking. Reducing the number of gravid females may also result in reduced transmission rates of pathogens. The combined effects of reduced oviposition and adult mortality could result in higher control potential of these microbial agents.     

家蝇卵巢中的信息化学物质引诱雌蝇产卵的研究

产卵期家蝇MuscadomesticaL .卵巢中的信息化学物质诱引家蝇产卵。行为试验表明 ,雌蝇产卵时优先选择被成熟卵巢污染过的发酵麦麸 ,诱集的产卵百分率依次为 :发酵麦麸 + 1对卵巢 >湿麦麸+ 1对卵巢 >发酵麦麸 >湿麦麸 ;卵巢提取物同样对产卵雌蝇有诱集活力。产卵雌蝇卵巢正己烷、二氯甲烷以及乙酸乙酯的提取物均能诱集雌蝇产卵 ,诱集的产卵百分率依次为 :雌蝇卵巢正己烷提取物 >乙酸乙酯提取物 >二氯甲烷提取物。推测家蝇卵巢中的信息化学物质可能含有家蝇产卵聚集信息素的组分。     

Identification of the best ovitrap installation sites for gravid Aedes (Stegomyia) aegypti in residences in Mirassol, state of São Paulo, Brazil

This study aimed at identifying the best ovitrap installation sites for gravid Aedes aegypti in Mirassol, state of S?o Paulo, Brazil. Ovitraps were installed in ten houses per block over ten blocks. Four ovitraps were placed per residence, one in the bedroom, one in the living room, and two outdoors with one in a sheltered area and one in an outside site. Each week for eleven weeks, visits were made to examine the ovitraps and to change the paddles used for egg-laying. Eggs were analyzed according to the trap location. The results showed that the outdoor sites received significantly more oviposition than indoor sites. Additionally, in respect to the outdoor sites, the outside site received significantly more oviposition than the sheltered site. A strong correlation was observed between positive traps and egg numbers. The results are discussed with respect to the best installation site of the traps and their implications in surveillance and control of dengue vectors.     

Oviposition bioassay responses of Culex tarsalis and Culex quinquefasciatus to neem products containing azadirachtin

Neem (Azadirachta indica A. Juss) products have been shown to exert pesticidal properties against a variety of insect species. In mosquito control programs, such products may have the potential to be used successfully as larvicides. In exploring other advantages of neem products, we studied the oviposition responses of Culex tarsalis Coquillett and Cx. quinquefasciatus Say to two experimental azadirachtin (AZ) formulations, wettable powder Azad ^– WP10 (WP) and emulsifiable concentrate Azad ^– EC4.5 (EC). Gravid Cx. tarsalis exhibited a distinct preference for the neem suspension of the WP, where significantly more egg rafts were collected from the treatment than from the control. The minimum effective AZ concentration for this activity was 0.5 ppm. The aged suspensions from 1–7 days at 0.5 and 1 ppm AZ were more active in eliciting oviposition responses in Cx. tarsalis than the fresh preparations. This activity of the aged suspensions lasted up to 14 and 21 days at 0.5 and 1 ppm AZ, respectively. Negative ovipositional responses were indicated in the tests of the EC vs. Cx. tarsalis, as well as both neem formulations vs. Cx. quinquefasciatus. In the tests of the EC formulation, significantly less gravid females were trapped by oviposition cups in the treatment than in the control, and in the tests of the WP significantly less egg rafts were collected from the treatment than from the control. The minimum effective concentrations for oviposition avoidance activity were 5 ppm AZ for Cx. tarsalis and 10 ppm AZ for Cx. quinquefasciatus, which lasted up to 1 and 4 days for these two species respectively. Neem products potentially used as mosquito larvicides may have many additional benefits in mosquito control programs, the oviposition modification noted in the current studies is one such example.     

A behavioural and electrophysiological study of ovi position cues for Culex quinquefasciatus

Abstract. Both the synthetic oviposition pheromone, erythro-6-acetoxy-5-hexa-decanolide (0.01–80 μg) and a polluted water sample (0.01–80%) significantly increased oviposition by gravid female Culex quinquefasciatus Say (Diptera: Culicidae) in a laboratory assay against clean water. An additive effect resulted when 0.05 μg oviposition pheromone was combined with the polluted water dilution series. Undiluted, however, both the oviposition pheromone and the polluted water sample, alone and in combination, reduced the oviposition response to control levels. A previously reported oviposition site odour component, 3-methylindole, also significantly increased oviposition at 1 × 10^_8 to 1 × 10^-7g but at 1 × 10^_7g significantly less oviposition occurred than in control treatments. The biological activities of the compounds were additionally evaluated electrophysiologically by an EAG assay. Female mosquitoes were electrophysiologically sensitive to the oviposition cues, both pheromone and habitat-related; EAG responses of males were considerably lower than of females with 3-methylindole.     

Odour-mediated upwind flight of Culex quinquefasciatus mosquitoes elicited by a synthetic attractant: a reappraisal

Abstract. Pile et al. (1991) report that the putative oviposition attractant ace-toxyhexadecanolide has four effects on gravid females of Culex quinquefasciatus Say: (1) it induces upwind anemotaxis, (2) near to an oviposition site it evokes an orthokinetic slowing of flight speed plus (3) a klinokinetic increase in turning rate, and (4) it causes females to stay longer at oviposition sites. This reappraisal shows that the first two of these conclusions are supported by the evidence provided but the second two are not.     

Protein hydrolysates and associated bacterial contaminants as oviposition attractants for the mosquito Culex quinquefasciatus

Abstract. Six protein or protein hydrolysate solutions were tested for activity as attractants for ovipositing Culex quinquefasciatus mosquitoes in the laboratory. Four of these solutions (egg albumin, lactalbumin hydrolysate, casein hydrolysate and yeast hydrolysate) were attractive to ovipositing females at varying concentrations, when compared to distilled water controls. Soy hydrolysate was repellent at 1%, but not significantly attractive or repellent at lower concentrations. 'Nulure', a tephritid fly bait containing protein hydrolysate, also had no significant effect on oviposition behaviour. Gravid females mostly oviposited within the first 4 h of the scotophase, regardless of the presence or absence of an oviposition attractant.
Lactalbumin hydrolysate 1% solution, with or without 0.1% neomycin antibiotic, was attractive to Cx quinquefasciatus . This effect was reduced by the presence of neomycin which, alone, had no effect on oviposition. Hence both lactalbumin hydrolysate and bacterial contaminants were shown to be attractive to gravid Cx quinquefasciatus .     

Recognition and avoidance of previously laid egg masses by the oblique-banded leafroller (Lepidoptera: Tortricidae)

Investigations were conducted on oviposition by laboratory-reared oblique-banded leafrollers, Choristoneura rosaceana Harris. In paperbag oviposition chambers, females deposited their egg masses in a significantly nonrandom, uniform manner in 16 of 20 replicates. Inpetri dish oviposition chambers, gravid females were exposed to egg masses recently deposited by other females. As determined by the magnitude of the mean direction vector of the second egg mass in relation to the first, females detected and avoided egg masses laid by other females. In addition, they avoided waxed paper from which a recently oviposited egg mass had been removed. This phenomenon is most likely due to a pheromone laid down with each egg mass.     

Oviposition response of the mosquito,Culex quinquefasciatus to the secondary metabolite(s) of the fungus,Trichoderma viride

Secondary metabolites produced by Trichoderma viride, a deuteromycetes fungus, under submerged culture condition were formulated and evaluated for oviposition attractancy against gravid females of Culex quinquefasciatus mosquito. At a concentration of 10 g ml-1 the formulation showed remarkable attractancy with an oviposition active index (OAI) of +0.52. When the oviposition attractancy of the formulation was compared with a known oviposition attractant, p-cresol, both at 10 g ml-1, the former was found to be more attractive to result in 70% egg laying than the later with 30% egg laying. Thin layer chromatography fractions of the secondary metabolites showed that a fraction with Rf value of 0.88 was highly active as oviposition attractant with an OAI of +0.65. Further work on identification of the active principle(s) of the microbial formulation might lead to an oviposition attractant useful in mosquito vector management.     

Identification of Components of the Oviposition Aggregation Pheromone of the Gregarious Desert Locust,Schistocerca gregaria (Forskal)

Acetophenone and veratrole have been identified as two major behaviourally active components of the oviposition aggregation pheromone of the desert locust, Schistocerca gregaria. These compounds were identified from the volatiles of egg pod froth using gas chromatography-electroantennographic (GC-EAD) and gas chromatography-mass spectrometric (GC-MS) analysis. Both compounds were shown to elicit aggregation of gravid females in oviposition bioassays; however, they do not act synergistically. Both acetophenone and veratrole individually at optimum doses induced about 70% egg laying, a value similar to that evoked with froth volatiles.     

Isolation of oviposition pheromone from the eggs of the sandfly Lutzomyia longipalpis

Abstract. Semiochemical components of eggs of the sandfly Lutzomyia longipalpis (Diptera: Psychodidae) were separated by high performance liquid chromatography. HPLC fractions were examined quantitatively and qualitatively by gas chromatography (GC). A bioassay was used to determine the oviposition attraction of gravid L. longipalpis to each of the fractions separately and a peak responsible for the semiochemical activity was identified. Gravid flies were placed in individual oviposition tubes to determine if the peak of interest was an oviposition stimulant. The active semiochemical fraction attracted gravid flies for oviposition. Furthermore, egg laying was enhanced: gravid flies exposed to the pheromone oviposited earlier and laid more eggs than control flies. GC analysis indicated that 1200 eggs (2 days old) gave a yield of 12.75 ug of active pheromone. This fraction had similar HPLC and GC retention times to caryophyllene oxide, suggesting comparable polarity and molecular weight.     

Experience‐ and age‐mediated oviposition behaviour in the yellow fever mosquito Stegomyia aegypti (=Aedes aegypti)

In repeated behaviours such as those of feeding and reproduction, past experiences can inform future behaviour. By altering their behaviour in response to environmental stimuli, insects in highly variable landscapes can tailor their behaviour to their particular environment. In particular, female mosquitoes may benefit from plasticity in their choice of egg‐laying site as these sites are often temporally variable and clustered. The opportunity to adapt egg‐laying behaviour to past experience also exists for mosquito populations as females typically lay eggs multiple times throughout their lives. Whether experience and age affect egg‐laying (or oviposition) behaviour in the mosquito Stegomyia aegypti (=Aedes aegypti) (Diptera: Culicidae) was assessed using a wind tunnel. Initially, gravid mosquitoes were provided with a cup containing either repellent or well water. After ovipositing in these cups, the mosquitoes were blood‐fed and introduced into a wind tunnel. In this wind tunnel, an oviposition cup containing repellent was placed in the immediate vicinity of the gravid mosquitoes. A cup containing well water was placed at the opposite end of the tunnel so that if the females flew across the chamber, they encountered the well water cup, in which they readily laid eggs. Mosquitoes previously exposed to repellent cups became significantly more likely to later lay eggs in repellent cups, suggesting that previous experience with suboptimal oviposition sites informs mosquitoes of the characteristics of nearby oviposition sites. These results provide further evidence that mosquitoes modify behaviour in response to environmental information and are demonstrated in a vector species in which behavioural plasticity may be ecologically and epidemiologically meaningful.     

Cover Caption

Females of the common green blow fly, Lucilia sericata, ovipositing on rat carrion. When females oviposit in aggregations, even‐aged larval offspring develop faster and fewer are preyed upon. These benefits imply that females engage in coordinated, pheromone‐mediated oviposition behavior. Yet, new data show that oviposition site‐seeking females do not respond to pheromones but to semiochemicals associated with gravid or non‐gravid feeding flies, taking chances that resources are suitable for oviposition and that ovipositing flies are present (see pages 651–660). Image: Sean McCann.     

Effects of forced egg-retention in Aedes albopictus on adult survival and reproduction following application of DEET as an oviposition deterrent.

The insect repellent DEET (0.1% concentration), used as a mosquito oviposition deterrent in the laboratory, influenced the retention and maintenance of mature eggs by caged gravid female Aedes albopictus Skuse. This egg-retention mechanism could benefit survival because the gravid females were ultimately able to lay maintained eggs upon availability of water, but the length of forced egg-retention time reduced the number of eggs laid per female. Gravid females with retained eggs also laid a higher percentage of eggs that failed to tan, and this percentage increased with time duration of egg-retention. Percent egg hatch was not significantly affected by DEET when used as an oviposition deterrent; however, percent hatch was affected by time duration of egg-retention in both treated (exposed to DEET) and untreated (control) gravid females. The rate of egg hatch was considerably reduced after three weeks of retention; this reduction declined to zero for treated and control females at six and four weeks post-treatment, respectively. The fecundity and fertility of gravid female Ae. albopictus were affected by the time duration of forced egg-retention.     

Oviposition Deterrence Is Likely an Effect,Not a Mechanism,in the Leaf Beetle <Emphasis Type="Italic">Chrysophtharta bimaculata</Emphasis> (Olivier) (Coleoptera: Chrysomelidae)

Oviposition deterrence is common in many insects as an evolutionary mechanism to reduce subsequent larval competition. We investigated a suspected case of oviposition deterrence by the paropsine chrysomelid, Chrysophtharta bimaculata. In paired choice tests, gravid females were found to prefer ovipositing on host leaves without conspecific eggs, confirming the presence of an apparent oviposition deterrence mechanism. Washing egg batches in water, hexane, or ethanol did not change this preference, suggesting that a soluble marking pheromone was not involved. Furthermore, it is unlikely that a plant-derived oviposition deterring substance is produced as beetles showed no significant oviposition preference between leaves which had been oviposited upon, but then had the eggs removed, and those that had never been oviposited upon. In trials using artificial leaves and mimic egg batches, “leaves” with “egg batches” placed near the tip of the leaf (the preferred site of oviposition in this species) were significantly less likely to be laid upon than artificial leaves where mimic eggs were placed away from the tip. In combination, the results strongly infer that oviposition deterrence in C. bimaculata is due to the mechanical blocking of the oviposition site by the first laid egg-batch, rather than a specific oviposition deterring cue. The apparent oviposition deterrence in this insect may well be an outcome or evolutionary effect of oviposition-site selection, rather than a clear adaptive mechanism to decrease larval competition.     

Improving efficacy of Box gravid traps for collecting Culex quinquefasciatus.

In outdoor screen cages, one and two-choice assays were conducted with gravid Cx. quinquefasciatus to assess and improve the efficacy of the Box gravid trap. Subsequently, in a field trial, a modified Box gravid trap was compared with three other gravid traps: the CDC gravid trap, the CFG gravid trap, and a novel design of a sticky gravid trap. A major disadvantage of the Box gravid trap, a relatively low trapping efficacy, was overcome with a simple modification to the trap configuration. By replacing the solid lid of the collecting tray with mosquito netting, the air velocity through the trap inlet was doubled and the quality of the resting sites for caught females was improved. As a result, the modified Box gravid trap caught and retained twice as many Cx. quinquefasciatus as its predecessor in two-choice bioassays in the screened cages and proved as effective as the commercially available CDC gravid trap in the field, but without damaging the collected specimens. Captures with the Box and CDC gravid traps were significantly greater than those of either the Counter Flow Geometry gravid trap or a sticky gravid trap.     

Aedes albopictus (Diptera: Culicidae) oviposition response to organic infusions from common flora of suburban Florida

We evaluated the oviposition response of gravid Aedes albopictus (Skuse) to six organic infusions. Laboratory and field‐placed oviposition cups baited with water oak (Quercus nigra L.), longleaf pine (Pinus palustris P. Mill), or St. Augustine grass (Stenotaphrum secundatum (Walt.) Kuntze), as well as binary infusion mixtures of each, were used. In addition, a triple‐cage, dual port olfactometer was used to measure upwind response of gravid individuals to these infusions. We found that Ae. albopictus deposited more eggs in infusion‐baited cups compared with water alone. Moreover, significantly more eggs were laid in the water oak and a water oak‐pine mixture as compared with the St. Augustine grass infusion in laboratory bioassays. However, a negative upwind response was observed with longleaf pine infusion in the olfactometer. In field cages, significantly more eggs were deposited in infusion‐baited cups as compared with water alone and a greater percentage of eggs were deposited in cups containing a water oak and the water oak‐longleaf pine mixture as compared with cups containing single infusions or their mixtures.