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.

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.

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.

Host cues induce egg hatching and pre-parasitic foraging behaviour in the mosquito parasitic nematode, Strelkovimermis spiculatus.

The responses of eggs in diapause and the infective stage of the nematode, Strelkovimermis spiculatus, to larvae of its host, Culex pipiens pipiens, were investigated in the laboratory. The results indicated that the presence of the host induced the egg hatching. The hatching rate increased when larger numbers of host larvae were present. Second instar mosquito larvae induced significantly higher hatching rates than any other stages. These findings explain how S. spiculatus synchronizes its life cycle with its host life cycle and population dynamics to increase its fitness when the natural habitat is constantly covered by water. Direct exposure of the nematode eggs to host larvae resulted in consumption of as many as 20 eggs per host. The eggs consumed caused 0-70% host mortality depending on the number consumed, which indicated an infection path other than cuticle penetration although it may represent a rare situation in nature. The result of host cue assays showed that the combination of chemical cues and physical vibration induced the highest egg hatching, which may increase the chance of host availability after hatching. However, once hatched, the nematodes ignored vibrations and used only chemical cues for host location. These findings suggest that eggs hatch synchronously with the most susceptible mosquito stage and with peak mosquito larval density.

Efficacy of Duet dual-action adulticide against caged Aedes albopictus with the use of an ultra-low volume cold aerosol sprayer.

Duet was field evaluated against caged Aedes albopictus, with the use of a truck-mounted ultra-low volume (ULV) cold aerosol sprayer at the rate of 90.6 ml/ha. Cages were placed in 3 rows stationed 30.5, 61.0, and 91.5 m downwind of the spray vehicle. Initial knockdown was >99% with mortality ranging between 95.8% and 98.0% across the distances. Volume median diameter (VMD) and droplet density were slightly reduced (16.4% and 18.6%, respectively) by distance. Reduction in VMD and droplet density, however, did not reduce mortality. The ULV application of Duet is an effective adulticide against Ae. albopictus and may be a useful tool for use in mosquito abatement programs.

Morphological sexual dimorphism in three species of anopheline mosquito larvae.

Sexual separation at the larval stage in anopheline mosquitoes with the naked eye is difficult. We have identified distinguishing spots visible to the naked eye on the 6th abdominal segment of 3rd and 4th instars of Anopheles stephensi, An. culicifacies, and An. subpictus. Based on this feature, male and female larvae can be differentiated morphologically at 3rd and 4th instars of these species. This is the first report on these characteristic spots that may have a wide application for larval sexing in mosquito taxonomy, physiology, toxicology, genetics, and control.