Surveillance of mosquitoes harnessing their buzzing sound.

Mosquito surveillance for vector-borne disease management relies on traditional morphological and molecular techniques, which are tedious, time-consuming, and costly. The present study describes a simple and efficient recording device that analyzes mosquito sound to estimate species composition, male-female ratio, fed-unfed status, and harmonic convergence interaction using fundamental frequency (F0) bandwidth, harmonics, amplitude, and combinations of these parameters. The study examined a total of 19 mosquito species, including 3 species of Aedes, 7 species of Anopheles, 1 species of Armigeres, 5 species of Culex, 1 species of Hulecoetomyia, and 2 species of Mansonia. Among them, the F0 ranges between 269.09 ± 2.96 Hz (Anopheles culiciformis) to 567.51 ± 3.82 Hz (Aedes vittatus) and the harmonic band (hb) number ranges from 5 (An. culiciformis) to 12 (Ae. albopictus). In terms of species identification, the success rate was 95.32 % with F0, 84.79 % with F0-bandwidth, 84.79 % with harmonic band (hb) diversity, and 49.7 % with amplitude (dB). The species identification rate has gone up to 96.50 % and 97.66 % with the ratio and multiplication of F0 and hb, respectively. This is because of the matrices that combine multiple sound attributes. Comparatively, combinations of the amplitude of the F0 and the higher harmonic frequency band were non-significant for species identification (60.82 %). The fed females have shown a considerable increase in F0 in comparison to the unfed. The males of all the species possessed significantly higher frequencies with respect to the females. Interestingly, the presence of male-female of Ae. vittatus together showed harmonic convergence between the 2nd and 3rd harmonic bands. In conclusion, the sound-based technology is simple, precise, and cost-effective and provides better resolution for species, sex, and fed-unfed status detection in comparison to conventional methods. Real-time surveillance of mosquitoes could potentially utilize this technology.

Genetic differentiation among Aedes aegypti populations from different eco-geographical zones of India.

The present study explicitly evaluated the genetic structure of Aedes aegypti Linn, the vector of dengue, chikungunya, and Zika viruses, across different geo-climatic zones of India and also elucidated the impact of ecological and topographic factors. After data quality checks and removal of samples with excess null alleles, the final analysis was performed on 589 individual samples using 10 microsatellite markers. Overall findings of this study suggested that, Ae. aegypti populations are highly diverse with moderate genetic differentiation between them. Around half of the populations (13 out of 22) formed two genetic clusters roughly associated with geographical regions. The remaining nine populations shared genetic ancestries with either one or both of the clusters. A significant relationship between genetic and geographic distance was observed, indicating isolation by distance. However, spatial autocorrelation analysis predicted the signs of long-distance admixture. Post-hoc environmental association analysis showed that 52.7% of genetic variations were explained by a combination of climatic and topographic factors, with latitude and temperature being the best predictors. This study indicated that though overall genetic differentiation among Ae. aegypti populations across India is moderate (Fst = 0.099), the differences between the populations are developing due to the factors associated with geographic locations. This study improves the understanding of the Ae. aegypti population structure in India that may assist in predicting mosquito movements across the geo-climatic zones, enabling effective control strategies and assessing the risk of disease transmission.

Ecological variations in adult life table attributes of Aedes aegypti (L.) from the desert and coastal regions of India.

The ecological variation in biological and adult life-table attributes of two populations of Aedes aegypti (Diptera: Culicidae) from the desert (Jodhpur) and coastal (Kolkata) regions of India are assessed to understand the reproductive and survival strategies. The results showed that females lived longer than males in both strains. The desert strain was more r-strategist because of its higher intrinsic rate of increase (rm  = 0.23), finite rate of increase (λ = 1.25), lower life expectancy of males (7.9 days) and females (14.4 days), mean generation time (T = 19.2 days) and doubling time (DT = 3.0 days). However, there was no difference in net reproductive rate (R0 ) between the desert and coastal strains. The coastal strain showed a longer female life expectancy (22.0 days) than the desert strain (14.4 days). However, the fecundity (eggs/female/day) was lower in the coastal strain (11.4) than in the desert strain (15.1). Conclusively, the desert (Jodhpur) strain is adapted to a better r-strategy than the coastal (Kolkata) strain of Ae. aegypti, which might be helpful to flourish in harsh environmental conditions. This study may provide accurate predictions of Ae. aegypti population dynamics for vector management.

Life table, survival and fecundity parameters of Aedes albopictus (Diptera: Culicidae) strains from desert and coastal regions of India.

The Asian tiger mosquito, Aedes albopictus (Skuse, 1895) is the anthropophilic and day-biter mosquito, residing in peridomestic habitats and is responsible for vector-borne diseases, i.e., chikungunya and dengue. The knowledge of mosquito life demographics is very important to provide a foundation for a successful vector control program. In the present study, a comparative adult life table, survival, and fecundity parameters were evaluated on desert (Jodhpur, Rajasthan) and coastal (Kolkata, West Bengal) strains of Aedes albopictus (Diptera: Culicidae) from India under the standard laboratory conditions. The results showed significant differences between these strains for the life expectancy of males, fecundity (total eggs/cohort and eggs/female life span), oviposition days, intrinsic rate of increase (rm), finite rate of increase (λ), and doubling time (DT). Female mosquitoes lived longer than the males in both strains. Significant differences were observed in the net reproductive rate (R0) between Kolkata and Jodhpur strains. The coastal (Kolkata) strain showed a longer male life expectancy (5.6 days) than the desert strain (4.0 days). However, the fecundity (eggs/female lifespan) was higher in Kolkata (360.2) than the Jodhpur strain (229.7). The Kolkata strain showed higher R0, rm, λ, and the lower T, DT than the Jodhpur strain. Conclusively, the results indicate that the coastal strain of Ae. albopictus is more r-strategist than the desert strain. This study may help in the precise prediction of Ae. albopictus population dynamics surviving in these geographical areas for vector management.

Prevalence of mouthpart sensilla and protease producing symbiotic gut bacteria in the forensic fly Chrysomya megacephala (Fabricius, 1794): Insight from foraging to digestion.

The blow fly, Chrysomya megacephala (Fabricius, 1794) is a globally prevalent forensically important species that helps to estimate accurate postmortem interval since the death. This fly occasionally causes cutaneous myiasis and transmits several pathogenic bacteria. To understand their ability of corpse detection and digestion of protein-rich meal, the present study describes the mouthpart sensilla and assessment of protease producing symbiotic gut bacteria. Scanning electron microscopy (SEM) showed the prevalence of trichoid sensilla (Tr), basiconic sensilla (Ba) and microtrichia (Mr) on labellar lobes, haustellum and maxillary palps of mouthparts. Bacterial particles of both rod (small and large) and spherical shaped were detected in the gut of C. megacephala using SEM. The bacterial density was higher on the foregut and midgut in comparison to the hindgut. From 72 bacterial isolates, 10 isolates from the foregut region showed considerable protease-producing efficacy ranging between 3.98 - 6.83 GHR and 9.73 - 34.68 U/ml protease. Among these, the most promising protease-producing bacterial isolate showed 16S rDNA sequence similarity (99.85%) with Chryseobacterium artocarpi DNA. This bacterium was the first report from flies. The findings of the study might help in better understanding of the role of sensilla in host perception and foregut symbiotic bacterial association in protein digestion in C. megacephala.

A membrane and blood-free approach to rear adult Aedes albopictus.

Most artificial dietary systems for feeding mosquitoes require a membrane feeder, host cues, phosphate saline buffers and a phagostimulant. These multicomponent feeders are complex, expansive and cumbersome that requires fully trained personnel. The objective of the present is to develop a simple sugar assisted protein (SAP) diet for the egg production of Aedes albopictus. The present study assessed the potential use of SAP dietary system on the engorgement, fecundity, preference of diet components and production of multiple generations of Ae. albopictus. Our data show that the female mosquitoes have strongly preferred a diet with: (i) a combination of sugar and protein over the individual component, and (ii) water over PBS (phosphate buffered saline) buffer as a carrier, whereas adenosine triphosphate (ATP) was not required as a phagostimulant. Based on our optimization data, the SAP diets (10-20% bovine serum albumin in 5% sucrose aqueous solution) do not require chemo-attractive lure, phagostimulant ATP, temperature and membrane feeding components. Female mosquitoes readily engorge on SAP diets and show similar rates of survival and fecundity compared to those when blood-fed on live animals. In addition, the number of eggs produced by female mosquitoes fed on SAP diets kept consistent for 10 consecutive generations. Our results indicate that SAP diet is a potential alternative against blood feeding that is simple and cost-effective diets for Ae. albopictus colony maintenance and to support large scale mass- production for experimental and other purposes.

The development of autonomous unmanned aircraft systems for mosquito control.

We constructed an electric multi-rotor autonomous unmanned aerial system (UAS) to perform mosquito control activities. The UAS can be equipped with any of four modules for spraying larvicides, dropping larvicide tablets, spreading larvicide granules, and ultra-low volume spraying of adulticides. The larvicide module sprayed 124 µm drops at 591 mL/min over a 14 m swath for a total application rate of 1.6 L/ha. The tablet module was able to repeatedly deliver 40-gram larvicide tablets within 1.1 m of the target site. The granular spreader covered a 6 m swath and treated 0.76 ha in 13 min at an average rate of 1.8 kg/ha. The adulticide module produced 16 µm drops with an average deposition of 2.6 drops/mm2. UAS pesticide applications were made at rates prescribed for conventional aircraft, limited only by the payload capacity and flight time. Despite those limitations, this system can deliver pesticides with much greater precision than conventional aircraft, potentially reducing pesticide use. In smaller, congested environments or in programs with limited resources, UAS may be a preferable alternative to conventional aircraft.

Large-Scale Operational Pyriproxyfen Autodissemination Deployment to Suppress the Immature Asian Tiger Mosquito (Diptera: Culicidae) Populations.

Effective suppression of container-inhabiting Asian Tiger [Aedes albopictus (Skuse)] (Diptera: Culicidae) and yellow fever [Aedes aegypti (L.)] (Diptera: Culicidae) mosquitoes presents one of the most intractable problems for modern mosquito control. Traditional tools often fail to control populations of these mosquito species, and are prohibitively expensive or have negative environmental impacts. Novel approaches and tools are urgently needed for integrated container-inhabiting mosquito management programs. One of the most promising techniques is autodissemination. We present the results of a long-term large-scale study conducted in a temperate urbanized environment representing typical Ae. albopictus habitats. Three treatment sites with autodissemination stations and three nearby reference sites were monitored for eggs, immature, and adult mosquitoes over a period of 3 yr from 2014 to 2016. Elevated larval and pupal mortality of 12-19% on average was the most notable outcome in sentinel cups of the treatment sites. The number of eggs in the treatment sites was significantly reduced in 2014, but not in 2015 or 2016. Adult populations remained similar in treatment and reference sites throughout the study. The impact of autodissemination on mosquito populations was lower than reported by previous investigations. Technical and logistical problems associated with wider coverage and working in multiple urban neighborhoods contributed to reduced efficacy. Incorporating autodissemination with routine mosquito control operations and commercializing this methodology for general public use will require further research on combining this tool with other novel or conventional technologies.

Heterodissemination: precision targeting container Aedes mosquitoes with a cohabiting midge species carrying insect growth regulator.

BACKGROUND: Management of Aedes albopictus and Ae. aegypti is challenging in large part due to the cryptic nature of their larval habitats. Autodissemination, using conspecific species to transfer pesticide, is unable to provide proactive control. Here we report results from a new hypothesis, heterodissemination, wherein females of the cohabiting non-biting midge, Chironomus decorus, reared in the laboratory, treated with pyriproxyfen, and released to transfer lethal concentrations to shared mosquito larval habitats. RESULTS: Pyriproxyfen-impregnated oil and powder formulations were developed. The average payload for each female midge treated with oil followed by powder formulations was 5.07 ± 0.92 µg of active ingredient or 1660 times the median lethal concentration (LC50 ) for Ae. albopictus or Ae. aegypti in 200 mL of water. Subsequent residue analysis showed pyriproxyfen transference from chironomids, treated with oil formulation only, into water-holding containers up to 2.06 ppb or 171.7 times the LC50 . Releasing 20 laboratory reared and contaminated Chironomus decorus into a small room resulted in 80.42 ± 0.67% and 75.67 ± 3.14% Ae. albopictus pupal mortality in open and cryptic sentinel ovicups, respectively. Container water volumes ranging up to 4 L did not affect efficacy. In a large field cage, 90.3 ± 2.5% Ae. albopictus mortality was resulted from releasing 100 treated female midges. Releasing 400 contaminated midges into a residential backyard resulted in 74.3% pupal mortality in sentinel ovicups. CONCLUSIONS: Room, large field cage and field release trials demonstrated that adult midges reared and treated in the laboratory transfer highly lethal concentrations of pyriproxyfen to Ae. albopictus container habitats. Heterodissemination provides a potential approach for precision, proactive mosquito control, which may draw attention for further studies. © 2020 Society of Chemical Industry.

Evaluation of enhanced oviposition attractant formulations against Aedes and Culex vector mosquitoes in urban and semi-urban areas.

Surveillance is not only an important tool to assess the population dynamics of vector mosquitoes, but it can also be used to control vector-borne diseases. Mosquito vectors that belong to several genera such as Anopheles, Aedes, and Culex play a crucial role in the transmission of malaria, dengue, chikungunya, Zika, and elephantiasis diseases worldwide. We tested the efficacy of two commercial-grade oviposition attractant formulations that were developed for the container-inhabiting Aedes aegypti, Aedes albopictus, and Culex quinquefasciatus mosquitoes present in urban or semi-urban environments. These attractants can lure gravid females. Field trials were conducted in residential yards during a post-rainy season in September and October. Our data showed considerable efficacy for both attractants. Aedes-attractant collected 1.6-fold more larvae (101.2 ± 10.5 larvae/trap) than the control, and Culex-attractant collected 1.27-fold more larvae (151.2 ± 12.5 larvae/trap) than the control, resulting in 0.8 and 0.7 oviposition attraction indices (OAIs), respectively. Regression analysis indicated that the Aedes-attractant was more stable than the Culex-attractant. Location and time did not alter the efficacy of these attractants. Our experiment suggests that these attractants can be used for the development of species-specific gravid traps to detect, estimate, and control the mosquito population in urban and semi-urban areas.

Seasonal field efficacy of pyriproxyfen autodissemination stations against container-inhabiting mosquito Aedes albopictus under different habitat conditions.

BACKGROUND: Control of the container-inhabiting mosquito, Aedes albopictus, is difficult using conventional methods due to its selection of cryptic peri-domestic habitats. We evaluated whether autodissemination stations can deliver sufficient pyriproxyfen to sentinel containers to produce significant pupal mortality in different habitats such as competing oviposition sites, peri-domestic habitats, junkyards and tire piles. We also tested how far the pesticide could be transferred over a 200-m range. RESULTS: Autodissemination stations performed effectively for 8-12 weeks under field conditions. Pupal mortality was reduced in sentinel cups with high-competing oviposition habitats (5 versus 20) in isolated plots; however, similar results were not seen in residential areas. Increasing the number of stations per plot (from 1 to 4) enhanced the efficacy. Peri-domestic habitat trials showed the highest pupal mortality (50.4%) and site contamination with pyriproxyfen (82.2%) among the trials. Autodissemination stations were able to contaminate habitats in a junkyard (50.0%) and tire piles (40.2%). Pyriproxyfen was detected in sentinel cups up to 200 m from stations. Detection of pyriproxyfen by residue analysis (0.005-0.741 µg L-1 ) in field samples confirmed the transfer of the insect growth regulator. CONCLUSION: Autodissemination stations have shown promising potential as a novel pest management tool against container mosquitoes in field trials in different habitats confronted by mosquito control personnel. © 2017 Society of Chemical Industry.

Evaluation of Lambda-Cyhalothrin and Pyriproxyfen Barrier Treatments for Aedes albopictus (Diptera: Culicidae) Management in Urbanized Areas of New Jersey.

Mosquito control programs in the United States are still searching for best management practices to control the Asian tiger mosquito, Aedes albopictus (Skuse; Diptera: Culicidae). Most intervention methods for this species are either labor intensive (e.g., source reduction) or short-term (e.g., ultra-low-volume adulticiding). We investigated the effectiveness of barrier spray pesticide applications within urban and suburban residential yards in New Jersey as a control strategy using a before-after-control-impact (BACI) approach. Applications of Demand CSR pyrethroid (9.7% AI lambda-cyhalothrin) only or combined Demand CSR and Archer IGR insect growth regulator (1.3% AI pyriproxyfen) applications resulted in significant and similar decreases in adult mosquito abundance post-treatment ranging from 78 to 74% respectively, compared with the untreated control. Both insecticides exceeded the 70% reduction threshold considered as effective for Ae. albopictus control for 2 to 4 wk. However, applications of Archer IGR alone did not reduce adult mosquito abundance. The field study results were supported by laboratory no-choice bioassays using treated leaf foliage. Our study is the first data driven evidence of the residual efficacy of barrier pesticide applications in New Jersey with lambda-cyhalothrin that provided significant reductions in adult Ae. albopictus populations for an extended duration.

Effectiveness of autodissemination stations containing pyriproxyfen in reducing immature Aedes albopictus populations.

BACKGROUND: Aedes albopictus, the Asian tiger mosquito, is an aggressive, highly anthropophilic, day-biting mosquito with an expanding geographic range. Suppression of Ae. albopictus is difficult because of the abundance and prevalence of larval habitats within peridomestic environments, particularly cryptic habitats such as corrugated extension spouts, fence post openings, discarded food containers, etc. Because of the challenges of eliminating or treating larval habitats of this species, we tested an autodissemination concept to contaminate these habitats with the insect growth regulator pyriproxyfen. METHODS: Our study was conducted in the City of Trenton (Mercer County), New Jersey, USA (40°12'N, 74°44'W). We selected six hot spots, where five or more Ae. albopictus males or females were collected based on weekly trap surveillance. A trapping unit was a city block, approximately 0.8 ha (hot spot), where we deployed 26 to 28 autodissemination stations per treatment plot. To gauge efficacy, we deployed BGS traps, oviposition cups, and sentinel cups in treatment and control locations. RESULTS: We found a significant reduction in eggs (P < 0.0001) and larval populations (P < 0.0001) as a result of treatment. Pupal mortality, as determined through bioassays, was also significantly higher in the treatment sites (P < 0.0001). CONCLUSION: Our results clearly show the potential and unique use of the autodissemination stations to control immature Ae. albopictus in urban areas. Penetration of larvicides with existing methods are difficult to reach cryptic habitats, but the autodissemination approach, which exploits the oviposition behavior of the target pest, can be integrated into intervention programs. New tools are urgently needed to curb the expansion and public health implications of Ae. albopictus and other container-inhabiting species.

Targeting a Hidden Enemy: Pyriproxyfen Autodissemination Strategy for the Control of the Container Mosquito Aedes albopictus in Cryptic Habitats.

BACKGROUND: The Asian tiger mosquito, Aedes albopictus, is a vector of dengue, Chikungunya, and Zika viruses. This mosquito inhabits a wide range of artificial water-holding containers in urban and suburban areas making it difficult to control. We tested the hypothesis that female-driven autodissemination of an insect growth regulator could penetrate cryptic oviposition habitats difficult to treat with conventional insecticidal sprays. METHODOLOGY: Oviposition preferences of Ae. albopictus females for open and cryptic cups were tested in semi-field experiments. Two conventional larvicidal sprayers were tested to determine droplet penetration and larvicidal efficacy in open and cryptic habitats using Bacillus thuringiensis var. israelensis (Bti) in the field. Finally, the efficacy of pyriproxyfen autodissemination stations was assessed in cryptic and open cups in residential areas during 2013 and 2014. PRINCIPAL FINDINGS: Gravid females strongly preferred cryptic (53.1±12.9 eggs/cup) over open (10.3±4.3 eggs/cup) cups for oviposition. Cryptic cups showed limited droplet penetration and produced 0.1-0.3% larval mortality with a conventional backpack and low-volume sprays of Bti. The autodissemination stations effectively contaminated these cryptic cups (59.3-84.6%) and produced 29.7-40.8% pupal mortality during 2013-2014. Significant pupal mortality was also observed in open cups. CONCLUSIONS: The autodissemination station effectively exploits the oviposition behavior of wild gravid females to deliver pyriproxyfen to targeted oviposition habitats. Although the pupal mortality in cryptic cups was relatively lower than expected for the effective vector control. Autodissemination approach may be a suitable supporting tool to manage Ae. albopictus immatures in the cryptic habitats those are less accessible to conventional larvicidal sprays.

The Insect Growth Regulator Pyriproxyfen Terminates Egg Diapause in the Asian Tiger Mosquito, Aedes albopictus.

The Asian tiger mosquito, Aedes albopictus, is a highly invasive mosquito species that transmits chikungunya and dengue. This species overwinters as diapausing eggs in temperate climates. Early diapause termination may be a beneficial strategy for winter mosquito control; however, a mechanism to terminate the diapause process using chemicals is not known. We tested the hypothesis that a hormonal imbalance caused by the administration of juvenile hormone analog would terminate egg diapause in A. albopictus. We tested the insect growth regulator pyriproxyfen on all developmental stages to identify a susceptible stage for diapause termination. We found that pyriproxyfen treatment of mosquito eggs terminated embryonic diapause. The highest rates of diapause termination were recorded in newly deposited (78.9%) and fully embryonated (74.7%) eggs at 0.1 and 1 ppm, respectively. Hatching was completed earlier in newly deposited eggs (25-30 days) compared to fully embryonated eggs (71-80 days). The combined mortality from premature diapause termination and ovicidal activity was 98.2% in newly deposited and >98.9% in fully embryonated eggs at 1 ppm. The control diapause eggs did not hatch under diapausing conditions. Pyriproxyfen exposure to larvae, pupae and adults did not prevent the females from ovipositing diapausing eggs. There was no effect of pyriproxyfen on diapausing egg embryonic developmental time. We also observed mortality in diapausing eggs laid by females exposed to pyriproxyfen immediately after blood feeding. There was no mortality in eggs laid by females that survived larval and pupal exposures. In conclusion, diapausing eggs were the more susceptible to pyriproxyfen diapause termination compared to other life stages. This is the first report of diapause termination in A. albopictus with a juvenile hormone analog. We believe our findings will be useful in developing a new control strategy against overwintering mosquito populations.

Human cancer: is it linked to dysfunctional lipid metabolism?

BACKGROUND: Lipid metabolism dysfunction leading to excess fat deposits (obesity) may cause tumor (cancer) development. Both obesity and cancer are the epicenter of important medical issues. Lipid metabolism and cell death/proliferation are controlled by biochemical and molecular pathways involving many proteins, and organelles; alteration in these pathways leads to fat accumulation or tumor growth. Mammalian Krüppel-like factors, KLFs play key roles in both lipid metabolism and tumor development. SCOPE OF REVIEW: Substantial epidemiological and clinical studies have established strong association of obesity with a number of human cancers. However, we need more experimental verification to determine the exact role of this metabolic alteration in the context of tumor development. A clear understanding of molecules, pathways and the mechanisms involved in lipid metabolism and cell death/proliferation will have important implications in pathogenesis, and prevention of these diseases. MAJOR CONCLUSION: The regulatory role of KLFs, in both cell death/proliferation and lipid metabolism suggests a common regulation of both processes. This provides an excellent model for delivering a precise understanding of the mechanisms linking altered expression of KLFs to obesity and tumor development. GENERAL SIGNIFICANCE: Currently, mouse and rats are the models of choice for investigating disease mechanisms and pharmacological therapies but a genetic model is needed for a thorough examination of KLF function in vivo during the development of an organism. The worm Caenorhabditis elegans is an ideal model to study the connectivity between lipid metabolism and cell death/proliferation.

Point-source and area-wide field studies of pyriproxyfen autodissemination against urban container-inhabiting mosquitoes.

Autodissemination of insecticides is a novel strategy for mosquito management. We tested if contaminated Aedes albopictus (Skuse) mosquitoes from a small area treated with commercial formulations (79gm a.i. pyriproxyfen/ha) using conventional techniques, would disseminate pyriproxyfen over a wider area. Pyriproxyfen showed LC50=0.012 ppb for Ae. albopictus. Direct treatment and autodissemination efficacy was measured as a pupal mortality by conducting Ae. albopictus larval bioassay. A tire pile (n=100 tires) treated by backpack sprayer as a point-source treatment showed higher pupal mortality in 2010 (60.8% for week 0-6) than in 2011 (38.3% for week 0-6). The sentinel containers placed for autodissemination in four compass directions out to 200-400m from the tire pile showed 15.8% pupal mortality (week 1-6) in the first year, and 1.4% pupal mortality in the second year. No significant difference was detected among the distances and direction for pupal mortality. In area-wide treatments, vegetation was sprayed in checkerboard pattern (3.7% of 105ha) using backpack sprayer in 2010 and in strips (24.8% of 94ha) using truck-mounted ultra-low volume (ULV) sprayer in 2011. In both years, the area-wide direct treatment efficacy was lower (30.3% during 2010 and 5.3% in 2011) than point-source treatments. Autodissemination in area-wide plots was higher in 2010 (10.3%) than 2011 (2.9%). However, area-wide treatments were ineffective on field populations of Ae. albopictus as monitored by using BGS traps. We found accumulation of pyriproxyfen in the week 6 autodissemination containers in both experiments. The differences in autodissemination in 2010 and 2011 can be attributed to higher rainfall in the second year that may have eroded the pyriproxyfen from treatment surfaces and sentinel containers. Our study shows that ULV surface treatments of conventional formulation do not work for autodissemination. The effectiveness of pyriproxyfen in autodissemination may be improved by developing specific formulations to treat vegetation and tires that can load high doses on mosquitoes.

Dual-treatment autodissemination station with enhanced transfer of an insect growth regulator to mosquito oviposition sites.

BACKGROUND: The Asian tiger mosquito, Aedes albopictus (Skuse), transmits important arboviral diseases and displaces native species. This peridomestic mosquito deposits eggs in natural and artificial containers. Container larval habitats tend to be cryptic and, therefore, difficult to reach by conventional insecticide treatments. We have developed an autodissemination station that transfers the insect growth regulator, pyriproxyfen, from the station to oviposition sites. Mosquitoes visiting the station become contaminated with an oil sticker followed by pyriproxyfen powder before exiting. RESULTS: In a room (31 m(3) ) bioassay a single station consistently achieved 100% Ae. albopictus pupal mortality against 10 oviposition containers. In a greenhouse (200 m(3) ) assay with two stations and 12 oviposition cups, 91.7% of the cups had been contaminated as shown, and 57.1% pupal mortality was recorded. Pyriproxyfen transfer was also detected by visualizing mosquito 'tarsal prints' using a fluorescent dye. CONCLUSIONS: The oil and pyriproxyfen powder dual-treatment station enhanced autodissemination efficacy by increasing toxicant attachment and retention on contaminated females. The autodissemination station offers a targeted, less environmentally damaging approach to manage cryptic container species.

Ovicidal activity of three insect growth regulators against Aedes and Culex mosquitoes.

Interspecific variations in the susceptibility of freshly and embryonated eggs of Aedes albopictus, Ae. aegypti, Ae. atropalpus and Culex pipiens were tested against three classes of insect growth regulators (IGRs) including ecdysone agonist (azadirachtin), chitin synthesis inhibitor (diflubenzuron) and juvenile hormone analog (pyriproxyfen) at 0.001, 0.01, 0.1 and 1.0ppm concentrations. Egg hatching inhibition was dose dependent, the highest being at 1.0ppm concentration for freshly laid eggs of Ae. albopictus (pyriproxyfen: 80.6%, azadirachtin: 42.9% and diflubenzuron: 35.8%). Aedes aegypti showed lower egg hatching inhibition when exposed to pyriproxyfen (47.3%), azadirachtin (15.7%) and diflubenzuron (25.5%). Freshly laid eggs of Cx. pipiens were most susceptible to diflubenzuron. Aedes atropalpus eggs were tolerant to all three classes of IGRs. Embryonated eggs of Ae. albopictus, Ae. aegypti, Ae. atropalpus and Cx. pipiens were resistant to pyriproxyfen, azadirachtin and diflubenzuron than freshly laid eggs. The median desiccation time (DT50) of Ae. atropalpus eggs was maximum (5.1h) as compared to Ae. aegypti (4.9h), Ae. albopictus (3.9h) or Cx. pipiens (1.7h) eggs. Insignificant relationship between the rates of desiccation and egg hatching inhibition suggests other factors than physical providing eggs the ability to tolerate exposures to various IGRs. Egg hatching inhibition was due to the alteration in embryonic development caused by IGRs. Changes in the egg shell morphology and abnormal egg hatching from the side of the egg wall instead of operculum, was observed at higher concentrations of diflubenzuron. Morphological and physiological variations in eggs may be the key factor to influence the ovicidal efficacy of IGRs. The present data provide a base line for the improvement of the ovicidal efficacy of the insecticide and its formulation.

Effects of larval habitat substrate on pyriproxyfen efficacy against Aedes albopictus (Diptera: Culicidae).

Pyriproxyfen, a juvenile hormone analog insect growth regulator (IGR), is a recommended insecticide for the control of container-inhabiting mosquitoes. The effects of eight container substrates (three plastics, wood, tire rubber, clay pot, concrete, and glass) were studied on reduction of bio-efficacy of pyriproxyfen for Aedes albopictus (Skuse) in the laboratory. Insect growth regulator-bioassay of third instars showed significant increase of LC50 with some substrates compared with control probably because of pyriproxyfen adsorption to the substrate. The LC50 for the control substrate (glass mug) was 0.029 microg/liter. In comparison to the control, LC50 increased 150.1 times with tire rubber (4.354 microg/liter) and 8-10 times with wood and high-density polyethylene plastic containers. Slight increases of LC50 were detected for clay pots, polypropylene, and polystyrene plastics (1.9-2.7 times). Pyriproxyfen efficacy with concrete substrate was higher than with glass containers, which might be caused by high pH. The pH of water with wood (5.6) and concrete (9.8) substrates differed from the other substrates tested (6.3-7.1). There was no pupal mortality at pH 5-11 without pyriproxyfen; however, interaction of pH (5-11) with pyriproxyfen (0.05 microg/liter) showed additive effects at both low and high pH values. Different substrates have variable impacts on pyriproxyfen efficacy, which might be associated with the failure of larval control strategies for the container mosquitoes under certain field conditions.