Forced egg retention and oviposition behavior of malaria, dengue and filariasis vectors to a topical repellent diethyl-phenylacetamide.

Egg retention and oviposition behavior of four species of mosquito vectors viz., Anopheles stephensi, Aedes aegypti, Aedes albopictus, and Culex quinquefasciatus to a topical insect repellent diethyl-phenylacetamide (DEPA) at 0.1-1000 mg/L was investigated under laboratory conditions. Based on oviposition activity indices, DEPA demonstrated concentration dependent oviposition deterrent effect to A. stephensi (-0.18 to -0.97), A. aegypti (-0.18 to -0.91) and A. albopictus (-0.50 to -0.98) females. In contrast, positive oviposition response by C. quinquefasciatus (+0.39 and +0.70) was observed respectively at 0.1 and 1 ppm, while 10 ppm of DEPA on water received 50% lesser egg rafts than control. Gravid Culex females laid no egg rafts at 100 and 1000 ppm DEPA treated bowls effecting 100% oviposition deterrence. Test mosquito females deposited most of their eggs (> 90%) in the absence of repellent odour, while DEPA odour on water surface forced them to retain huge numbers of eggs. Females of A. aegypti, A. albopictus and A. stephensi retained 49, 67 and 50% of total eggs, respectively throughout the experiment. Egg retention by Culex females due to DEPA on the water surface was ca. 65%, equivalent to 4 egg rafts. Therefore, DEPA at lower concentrations could effectively disturb the oviposition by these vectors. Application of repellents in small water bodies would help in reducing the population build up of mosquitoes near human households and could be useful in the integrated management of mosquito vectors.

Behavioral and electrophysiological responses of Aedes albopictus to certain acids and alcohols present in human skin emanations.

Human skin emanations attract hungry female mosquitoes toward their host for blood feeding. In this study, we report the flight orientation and electroantennogram response of Aedes albopictus females to certain unsaturated acids and alcohols found in human skin. In the Y-tube olfactometer, odors of lactic acid and 2-methyl-3-pentanol attracted 54-65% of Ae. albopictus females at all doses in a dose-dependent manner. However, at the highest dose (10(-2) g), the acids repelled 40-45% females. Attractancy (ca. 62-68%) at lower doses and repellency (ca. 30-45%) at higher doses were recorded for 3-methyl-3-pentanol and 1-octen-3-ol, while 5-hexen-1-ol, cis-2-hexen-1-ol, and trans 2-hexen-1-ol odor repelled ca. 55-65% of Ae. albopictus females at all doses. Antenna of female Ae. albopictus exhibited a dose-dependent EAG response up to 10(-3) g of L-lactic acid, trans-2-methyl-2-pentenoic acid, 2-octenoic acid, trans-2-hexen-1-ol and 1-octen-3-ol stimulations; however, the highest dose (10(-2) g) caused a little decline in the EAG response. EAG response of 9-10-fold was elicited by lactic acid, 2-octenoic acid, trans-2-hexenoic acid, and 3-methyl-3-pentanol, while cis-2-hexen-1-ol and trans-2-methyl pentenoic acid elicited 1-5-fold responses compared to solvent control. A blend of attractive compounds could be utilized in odor-baited trap for surveillance and repellent molecules with suitable formulation could be used to reduce the biting menace of mosquitoes.

Olfaction in Asian tiger mosquito Aedes albopictus: flight orientation response to certain saturated carboxylic acids in human skin emanations.

The flight orientation response of nonblood-fed and hungry Aedes albopictus females was studied in a Y-tube olfactometer at 10(-6) to 10(-2) g odor plumes of saturated carboxylic acids (C1-C20), in which C2-C18 were the main constituents of human skin emanations. Thirteen acids viz C1, C2, C3, C5, C6, C8 C9, C10, C12, C14, C16, C18, and C20 showed attractance at odor plumes ranging from 10(-5) to 10(-3) g doses, while five acids viz C4, C7, C11, C15, and C19 showed repellence at 10(-4) to 10(-2) g to test mosquitoes. Tridecanoic acid (C13) showed attractance only at 10(-4) g dose while higher doses caused repellence. Dose-dependent reversal of orientation behavior from attractance to repellence was observed at 10(-2) g plumes of C5, C9, C10, C13, C17, C19, and C20 acids. The outcome of the study will help in the identification of odoriferous acids as potential attractants, repellents, or attraction inhibitors, which may find their application in the repellent formulations and odor-baited traps for surveillance and control of mosquitoes.

Oviposition and flight orientation response of Aedes aegypti to certain aromatic aryl hydrazono esters.

Aedes aegypti is a day-biting, highly anthropophilic mosquito and a potential vector of dengue and chikungunya in India. A. aegypti is a container breeder, generally oviposit in the stored and fresh water bodies, and discarded containers near residential areas that provide suitable habitats for oviposition by gravid females. The diurnal activity and endophilic nature of these mosquitoes have increased the frequency of contact with human being. Assured blood meal from human host in an infested area leads to increased disease occurrence. Gravid mosquitoes can potentially be lured to attractant-treated traps and could subsequently be killed with insecticides or growth regulators. In this direction, oviposition by A. aegypti females to aryl hydrazono esters (AHE)-treated bowls at 10 ppm concentration was tested in dual choice experiment, and their orientation response to these ester compounds was studied in Y-tube olfactometer. Among the esters tested, AHE-2, AHE-11 and AHE-12 elicited increased egg deposition with oviposition activity indices (OAI) of +0.39, +0.24 and +0.48, respectively, compared to control; in contrast, AHE-8, AHE-9 and AHE-10 showed negative oviposition response with OAI of -0.46, -0.35 and -0.29, respectively, at 10 mg/L. In the Y-tube olfactometer bioassay, AHE-2 attracted 60 % females compared to control, while to the odour of AHE-11 and AHE-12, about 70 % of the females were trapped in treated chambers. In contrast, only 27-30 % of gravid females entered the chamber releasing AHE-8, AHE-9 and AHE-10 odour plumes, while 70 % entered control chamber, evincing a possible non-preference of treatment odours as well as interference with olfactory receptors. These compounds have the potential for application as oviposition stimulants or deterrents for surveillance and control of mosquito population using ovitraps.

Gas-chromatography and electroantennogram analysis of saturated hydrocarbons of cruciferous host plants and host larval body extracts of Plutella xylostella for behavioural manipulation of Cotesia plutellae.

Saturated hydrocarbons (SHC) of five cruciferous host plants viz., cabbage, cauliflower, broccoli, knol khol and Brussels sprout and the larvae of diamondback moth (DBM), Plutella xylostella reared on these host plants were identified through gas-chromatography. The hydrocarbon profile of host plants and larval body extract of DBM reared on respective host plants revealed a wide variation in quantity as well as quality. Long chain hydrocarbons C26-C30 were detected in all the extracts. In electroantennogram (EAG) studies, SHCs at 10(-3) g dose elicited differential EAG response in the antennal receptors of gravid Cotesia plutellae females. Tricosane (C23) and hexacosane (C26) elicited 10-fold increased EAG response compared to control stimulus. Long chain hydrocarbons C27, C28 and C29 elicited, 6-7 fold increased responses. The sensitivity of antenna was 4-5 folds for C25, C14, C24, C15 and C30, while the short chain hydrocarbons elicited 2-3 fold increased EAG responses. Dual choice flight orientation experiments in a wind tunnel revealed that the gravid C. plutellae females preferred the odour of C16, C26, C29, C15, C21, C23, C30, C27, C24 and C22 as 60-70% females oriented and landed on SHC treated substrate compared to control odour, while the odour of eicosane (C20), pentacosane (C25) and octacosane (C28) were not preferred by the females.

Synthesis and bio-evaluation of aryl hydrazono esters for oviposition responses in Aedes albopictus.

A novel series of aryl hydrazono esters (AHE) (1-13) were synthesized (yield 76-98%) to study the oviposition responses in Aedes albopictus (Skuse) mosquitoes for the first time. At a concentration of 10µgml(-1) in dual choice experiment, among the screened compounds, AHE-12 showed remarkable oviposition attractant activity with an oviposition activity index (OAI) of +0.299 (greater than 95% confidence limit) comparable to p-cresol (OAI +0.320) which is well-reported oviposition attractant for Aedes aegypti. Conversely, AHE-10 exhibited highest oviposition deterrent activity with OAI -0.247. The possible utilization of these compounds will be in integrated vector management strategies.

Oviposition pheromones in haematophagous insects.

Pheromones influencing oviposition behavior in females of haematophagous insects have been the interest of recent past by many group of scientists working on oviposition pheromones. Finding and choosing a good site for oviposition is a challenging task for females of haematophagous insects, especially in those insects which does not have the parental care. Their decisions have far-reaching and profound consequences for the life history of the offspring. In such blood feeding insects, the choice of oviposition site is affected by pheromones, which may function either as deterrents or stimulants in short range, while they may also act as repellents or attractants in long range perception. During the location of a suitable oviposition site for egg laying or a potential host for blood feeding, haematophagous insects mainly use olfactory and visual cues. These pheromones are produced by the ovipositing female or by conspecific larvae co-occurring with gravid females. Adult females detect oviposition pheromones by odor receptors on the antennae, as well as by contact chemoreceptors on tarsi, mouthparts and antennae. Different cues exploited by gravid females from a diversified arena include egg, larva, habitat, microbes, infusions and plant produced volatiles influence the oviposition behavior. Traps baited with pheromones, infusions, and insecticides shall be promising tools for monitoring and control of target insect using integrated vector management strategies.

Electrophysiological, flight orientation and oviposition responses of three species of mosquito vectors to hexadecyl pentanoate: residual oviposition repellent activity.

Understanding on the chemical ecology of mosquito behavior is of paramount importance in developing control programs employing attractants and repellents. Several workers focused on topical repellents and oviposition attractants of mosquitoes, however, only limited work has been accomplished on mosquito oviposition repellents. The present systematic investigation provides evidence on the effectiveness of a C21 fatty acid ester- hexadecyl pentanoate, to stimulate antennal olfactory receptors of Aedes aegypti (L.), Ae. albopictus (Skuse), and Anopheles stephensi (Liston) that mediate their long-range olfaction guided flight orientation behavior by repelling the gravid females of these mosquito vectors in the olfactometer. The compound loaded onto an effervescent tablet retained its repellent property in the treated substrates for up to 1 wk at 10 mg/L. In places, where the mosquito breeding habitats are near to human habitations, could be treated with hexadecyl pentanoate to repel the ovipositing gravid females as a component of the integrated approach for mosquito management by disrupting the mosquito life cycle and population growth.

Surface morphology and morphometric analysis of sensilla of Asian tiger mosquito, Aedes albopictus (Skuse): an SEM investigation.

BACKGROUND & OBJECTIVES: The sensilla and sensory mechanism play a significant role in hostseeking and oviposition behaviour of mosquitoes, which enable them to transmit various diseases to humans. Aedes albopictus (Skuse) has emerged as a major vector of Chikungunya virus in the recent epidemics in most parts of southern India. Studies on the sensory structures of dengue vector, Aedes aegypti (Linn) are comprehensive; whereas information on the sensillary systems of Asian tiger mosquito, Ae. albopictus is inadequate. Therefore, the present study has been carried out to observe various types of sensilla located on the antenna, maxillary palp, labial palp, tarsi and ovipositor of Ae. albopictus using scanning electron microscopy. METHODS: The antennae, maxillary palpi, labellum, tarsi and ovipositor of 10 different female mosquito of Ae. albopictus were fixed individually in 2.5% glutaraldehyde solution, washed twice and dehydrated with ascending grades of ethanol. Samples were cleared with xylene, air-dried, mounted on stubs, gold coated in an ion-sputtering unit and the sensilla were viewed between 5 and 10 KV using FEI-Quanta 400-EDAX scanning electron microscope. ANOVA revealed significant differences in the morphometric features of various sensilla. RESULTS: In the antenna Sensilla trichoidea are numerously distributed in all flagellar segments revealed four distinct subtypes. Two types of grooved peg sensilla were observed. Sensilla coeloconica was observed in the terminal flagellum of antenna and tarsomeres with large variation in diameter. Sensilla chaetica are distributed throughout the body and revealed greater variation in morphology and morphometric parameters. INTERPRETATION & CONCLUSION: The significant difference among various types of sensilla would possibly reveal their functions. The porous sensilla are olfactory and contact chemoreceptors while the aporous sensilla would play the role of mechanoreception. Sensilla coeloconica on the antenna, tarsus showed major differences with Ae. aegypti. The ovipositor sensilla revealed three types of chaetica arranged in rows but has not been reported earlier with other mosquito species.

Mediation of oviposition responses in the malaria mosquito Anopheles stephensi Liston by certain fatty acid esters.

The chemical factors involved in oviposition site selection by mosquitoes have become the focus of interest in recent years, and considerable attention is paid to the chemical cues influencing mosquito oviposition. Studies on synthetic oviposition attractants/repellents of long-chain fatty acid esters against Anopheles stephensi are limited. Screening and identification of chemicals which potentially attract/repel the gravid females to/or from oviposition site could be exploited for eco-friendly mosquito management strategies. The ester compounds demonstrated their ability to repel and attract the gravid A. stephensi females in the treated substrates. Significant level of concentration-dependent negative oviposition response of mosquitoes to octadecyl propanoate, heptadecyl butanoate, hexadecyl pentanoate, and tetradecyl heptanoate were observed. In contrast, decyl undecanoate, nonyl dodecanoate, pentyl hexadecanoate, and propyl octadecanoate elicited concentration-dependent positive oviposition responses from the gravid mosquitoes. Forcing a female to retain her eggs due to unavailability of a suitable oviposition site and attracting them to lay the eggs in a baited ovitraps shall ensure effective control of mosquito breeding and population buildup because the oviposition bioassay target the most susceptible stage of an insect life cycle. Treating relatively smaller natural breeding sites with an effective repellent and placing ovitraps containing an attractant in combination with insect-growth regulator (IGR)/insecticide would be a promising method of mosquito management.

Electroantennogram, flight orientation, and oviposition responses of Aedes aegypti to the oviposition pheromone n-heneicosane.

Oviposition pheromones specifically influence the females of many insects to lay eggs in the sites resulting in more egg deposition. A previous report describes the principal role of n-heneicosane (C(21)) identified and characterized from the larval cuticle of Aedes aegypti (L.) in attracting the gravid mosquitoes to oviposit in treated substrates among other chemical components. However, the means by which this compound is perceived by the females for oviposition has not been reported. In this study, we have recorded the peripheral olfactory responses from the antenna of Ae. aegypti from 10(-7) g to 10(-3) g doses of n-heneicosane. The EAG response of female mosquitoes increased in a dose-dependent manner with increasing stimulus strength. In the orientation assay using Y-maze olfactometer, female mosquitoes were attracted to the odor plume of 10(-6) g and 10(-5) g dose, while the higher dose of 10(-3) g plume enforced repellency to gravid mosquitoes. The response to oviposition substrates by gravid Ae. aegypti females differed across the range of concentrations of n-heneicosane under multiple choice conditions, larger number of eggs were deposited in 10 ppm (10 mg/l) solutions compared to lower and higher concentrations indicating 10 ppm was most attractive. Application of n-heneicosane at 10 ppm in breeding habitats will be a useful method to attract the gravid mosquitoes using ovitraps for surveillance and monitoring. The possible use of this compound in monitoring of mosquito population in endemic areas in relevance to integrated vector management strategies is discussed in detail.

Oviposition responses of Aedes aegypti and Aedes albopictus to certain fatty acid esters.

Laboratory studies were carried out to observe the oviposition responses of Aedes aegypti (L.) and Aedes albopictus (Skuse) to several C21 fatty acid esters. The oviposition activity of these dengue and chikungunya vectors to the long-chain fatty acid esters of C21 length have not been reported earlier. From the multiple choice experiments on oviposition activity in standard mosquito cages, it was observed that compounds hexadecyl pentanoate, tetradecyl heptanoate and tridecyl octanoate presented significant oviposition repellent activity against the two mosquito species, while one compound propyl octadecanoate was found to attract A. aegypti to the treated oviposition substrate at 1- and 10-ppm concentrations. The possible utilization of these esters in integrated vector management is discussed.