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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.

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.

Conventional larvicide delivery strategies originally developed for permanent and floodwater mosquitoes have proved suboptimal in the small, scattered, and cryptic larval habitats preferred by container-inhabiting Aedes mosquitoes. New methods such as autodissemination, wherein adult mosquitoes spread insecticides to their own larval habitats, have been under study. Another novel delivery method termed heterodissemination, i.e. larvicide delivery by other species sharing the same habitats, has also been proposed. We conducted a proof-of-concept study with four independent experiments using American bullfrogs (Lithobates catesbeianus) and green frogs Lithobates clamitans as carriers of pyriproxyfen, an insect growth regulator, under semi-field conditions in three different locations, two in New Jersey, and one in Utah. Frogs with attached slow-release pyriproxyfen tablets were introduced into outdoor enclosures with water containers. Water samples from the containers were periodically tested using larval Aedes albopictus and Culex pipiens mosquitoes to assess mortality and percent eclosure inhibition. Overall pupal mortality [95% credible intervals] estimated by Bayesian analysis for the treatment group was 73.4% [71.3–75.2] compared to 4.1% [2.9–5.5] for the control group. Mortality within treatment groups in four different experiments ranged from 41 to 100%, whereas control mortalities ranged from 0.5% to 11%. We conclude that heterodissemination is a promising and effective approach deserving of further study.

Summary Background Dietary salts sodium (Na+), potassium (K+), magnesium (Mg2+), and calcium (Ca2+) are important in metabolic diseases. Yet, we do not have sufficient understanding on the salts global molecular network in these diseases. In this systematic review we have pooled information to identify the general effect of salts on obesity, insulin resistance and hypertension. Aims To assess the roles of salts in metabolic disorders by focusing on their individual effect and the network effect among these salts. Methods We searched articles in PubMed, EMBASE and Google Scholar. We selected original laboratory research, systematic reviews, clinical trials, observational studies and epidemiological data that focused on dietary salts and followed the preferred reporting items for systematic review in designing the present systematic review. Results From the initial search of 2898 studies we selected a total of 199 articles that met our inclusion criteria and data extraction. Alterations in metabolic pathways associated with the sensitivity of sodium, potassium, magnesium and calcium may lead to obesity, hypertension, and insulin resistance. We found that the results of most laboratory research, animal studies and clinical trials are coherent but some research outcome are either inconsistent or inconclusive. Conclusion Important of salts in metabolic disorder is evident. In order to assess the effects of dietary salts in metablic diseases, environmental factors, dietary habits, physical activity, and the microbiome, should be considered in any study. Although interest in this area of research continues to grow, the challenge is to integrate the action of these salts in metabolic syndrom.

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.

Integrated pest management (IPM) has recognized as a sustainable method for pest prevention, monitoring, and control. The purpose of this review article focused on biological control potential and challenges in Afghanistan. Biological control is beginning to increase in Afghanistan, in large part due to the establishment of graduate studies in entomology in 2008. Afghan farmers have restricted knowledge about agrochemicals and mostly use a few conventional chemical pesticides and fertilizers. The development and registration of biological agents as an alternative is a more recent movement. Only a single parasitoid agent and three microbial products were registered or pending registration for commercial use on various crops. Products based on Trichogramma brassicae (Bezdenko 1968) for lepidopteran pests, Trichoderma viride against soil-borne pathogens including Rhizoctonia solani, Macrophomina phaseolina, and Fusarium species are most widely spread. Also, T. viride, Cydia pomonella granulovirus, and Helicoverpa zea nucleopolyhedrovirus registered in 2015. Pesticide registration is the responsibility of the Afghan Ministry of Agriculture, Irrigation and Livestock, with the assistance of the Pesticides Division of Plant Protection and Quarantine Department in coordination with the Ministry of Public Health. The pesticide management board of the National Environmental Protection Agency has not yet signed the membership of the World Trade Organization (WTO) or the International Plant Protection Convention (IPPC). Currently, only one foreign company, the National Horticulture and Livestock Project (NHLP), produces microbial pesticides. However, it promised that two more other companies, the Center for Agricultural Bioscience International and the Afghanistan Agriculture Inputs Projects, will start production shortly. This is a review of the brief history of biological control of pests, with a view on current challenges of pest control practices as well the status of the market and the registration procedures for biocontrol agents, along with challenges and opportunities for the development of biocontrol in Afghanistan.

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.

BACKGROUND Given the lack of vaccines for most vector-borne diseases, vector control is often the primary option for disease control. Aedes albopictus are difficult to control because the immatures primarily develop in containers ubiquitous in residential properties. Conventional adulticide campaigns often result in brief, rebounding population declines, so incorporating new techniques into an integrated pest management program is imperative. We performed combined area-wide applications of the larvicides Bacillus thuringiensis var. israelensis and pyriproxyfen with the adulticide sumithrin and prallethrin to achieve extended suppression of Ae. albopictus populations in Trenton, NJ, USA. We deployed bioassay cups to assess the spatial penetration and efficacy of the applications. RESULTS Inhibition of adult emergence was significantly higher in the treatment bioassay cups than in laboratory controls (z = 4.65, P < 0.0001) and field control bioassay cups (z = 8.93, P < 0.0001). We observed a lower trend in adult numbers following season-long combined application of pyriproxyfen and adulticide, with numbers of adult Ae. albopictus at the treatment site up to five times lower than at the control site. CONCLUSION Pyriproxyfen is a powerful mosquito larvicide and pupacide with low mammalian toxicity that shows promise for area-wide vehicle-mounted (either ground or airborne) applications. © 2018 Society of Chemical Industry.

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

To understand the infection strategies of the mosquito parasitic mermithid nematodes Romanomermis iyengari (Welch, 1964) and Strelkovimermis spiculatus (Poinar and Camino, 1986), we have investigated the foraging behaviors of the pre-parasites of both species. Romanomermis iyengari with body length of 0.99 ± 0.071 mm (twice as S. spiculatus 0.55 ± 0.01 mm) swims faster (2.76 ± 0.13 cm/min) than S. spiculatus (1.61 ± 0.09 cm/min). The nematode pre-parasites of R. iyengari showed negative phototaxis in responding to natural light, which caused them to migrate to shade where mosquito larvae tend to aggregate. This behavior helped the nematode to gain significantly higher parasitism when shade was present (37.7 ± 2.74%) than no shade (11.3 ± 0.95%). In contrast, S. spiculatus pre-parasites had no response to natural light and therefore, shade contributed no parasitism gain to this species. The host searching pattern data indicated that the two species used different strategies for host searching. The pre-parasites of R. iyengari tended to do long range search and responded to host aggregation habitat but slowed down and do more localized search when host present. However, the species of S. spiculatus , tended to do localized movement when hosts were absent regardless shade. Once host larvae present in nearby location, the nematodes responded to the host cue and moved faster toward their hosts. The active foraging behavior of R. iyengari may suggest that the species is more suitable for mosquito control in clear and moving water such as streams. However, the passive stay and wait behavior of S. spiculatus might be more suitable against mosquitoes in small sized water bodies such as temporarily flooded or stagnant ponds.

The mosquito-parasitic nematode, Strelkovimermis spiculatus (Mermithidae: Nematoda) emerges from hosts and aggregates to form mating clusters characterized by intense male-male competition for females. Successful males deposit an adhesive copulatory plug over the female vulva after insemination. In choice experiments, males strongly preferred virgin females, whereas plugged females were ignored. Males made no attempt to displace copulatory plugs deposited by previous males. Plugged females repelled males without the need for physical contact. The observed chemical repellency was independent of females, since excised plugs alone showed the same negative male response. Removal of the plug shortly after mating reduced fecundity by 90%, presumably due to spermatids leaking from the vulva. The plug as a nutritional gift hypothesis was rejected because there was no post-mating reduction in plug size that would have indicated absorption. Despite being a male adaption to sperm competition, we conclude that the copulatory plug serves the fitness needs of both males and females in multiple roles that include paternity preservation, reduced male harassment, improved male search for mates, increased fecundity, and sperm retention.

We examined manipulation of mosquito behavior by the parasitic mermithid nematode, Strelkovimermis spiculatus. This nematode species typically infects early instar host larvae and emerges after parasitic development to kill last-instar larvae. Parasitized adults, however, have occasionally been reported from field collections. We obtained low rates (1.7-11.5%) of parasitized adults in laboratory exposures only when Culex pipiens pipiens fourth-instar larvae nearing pupation were exposed to infective nematodes. This did not allow an adequate interval for parasitic development in immature host stages. Parasitized adult females in a multiple-choice assay were three times more likely to seek water than a blood source (63.1 vs. 20.5%), whereas uninfected females were twice as likely to seek blood than water (64%3.9 vs. 32.6%). This altered host behavior benefits the parasite by providing the only mechanism for dispersal and colonization of new host habitats while concurrently avoiding risks from the defensive behaviors associated with blood-feeding. Behavioral alternation in Cx. p. pipiens larval hosts was also examined using larvae infected as second instars to allow for a normal duration of parasitic development. As larvae neared pupation and parasite emergence, parasitized larvae became more spatially aggregated than unparasitized larvae. This altered host behavior benefits the parasite by providing a corresponding increase in post-parasite aggregation, which facilitates formation of large mating clusters and concomitantly reproductive success. Parasites derive fitness gains by overriding host autonomy, whereas hosts have zero fitness once parasitism is established, suggesting a coevolutionary response is inoperative and that the behavioral modifications may be adaptive.

Barrier applications, treating vegetation and other potential mosquito resting areas with residual pesticides, have become standard practice for commercial pest management professionals offering mosquito control services. These treatments are generally effective in reducing numbers of the Asian tiger mosquito, Aedes albopictus (Skuse). In the temperate climates, adult Ae. albopictus populations are naturally eliminated each winter and must be grown from diapausing eggs the following spring. Therefore, early-season control interventions timed before populations rebound may further reduce population growth compared with interventions that take place after the population has peaked. We compared Early (1 June) barrier applications of lambda-cyhalothrin (Demand CS) mixed with the insect growth regulator pyriproxyfen (Archer IGR) to Late applications (31 July) in urban and suburban residential parcels in New Jersey. These applications resulted in 43 and 51% reductions in adult Ae. albopictus populations in the Early and Late intervention study parcels, respectively, when compared with the untreated Controls. Indirect applications of pyriproxyfen (i.e., rainwater runoff transferring pesticide) to potential larval habitat did not cause mortality in laboratory larval bioassays but direct mist applications of the Archer and Demand combination to larval habitats led to elevated larval mortality over a 6-wk period. The Early-season intervention offered no advantage in efficacy on adult populations compared with the Late-season intervention, which targeted peak mosquito populations. Our study provides support for the efficacy of barrier applications and illustrates the importance of timing barrier treatments to coincide with established Ae. albopictus action thresholds.

Given the lack of vaccines for most vector-borne diseases, vector control is often the primary option for disease control. Aedes albopictus are difficult to control because the immatures primarily develop in containers ubiquitous in residential properties. Conventional adulticide campaigns often result in brief, rebounding population declines, so incorporating new techniques into an integrated pest management program is imperative. We performed combined area-wide applications of the larvicides Bacillus thuringiensis var. israelensis and pyriproxyfen with the adulticide sumithrin and prallethrin to achieve extended suppression of Ae. albopictus populations in Trenton, NJ, USA. We deployed bioassay cups to assess the spatial penetration and efficacy of the applications.Inhibition of adult emergence was significantly higher in the treatment bioassay cups than in laboratory controls (z = 4.65, P0.0001) and field control bioassay cups (z = 8.93, P0.0001). We observed a lower trend in adult numbers following season-long combined application of pyriproxyfen and adulticide, with numbers of adult Ae. albopictus at the treatment site up to five times lower than at the control site.Pyriproxyfen is a powerful mosquito larvicide and pupacide with low mammalian toxicity that shows promise for area-wide vehicle-mounted (either ground or airborne) applications. © 2018 Society of Chemical Industry.

Arboviruses diseases, especially dengue, chikungunya and Zika are responsible for millions of human sickness and a significant number of deaths worldwide. There is no effective treatment and cure available for these arboviral diseases. These diseases are transmitted mainly by Aedes mosquitoes such as Aedes aegypti and Aedes albopictus. They are highly anthropophilic, container-inhabiting and diurnal mosquitoes surviving in peridomestic habitats. The management of dengue, chikungunya and Zika diseases mainly depends on vector management, but the habit and habitats of these mosquitoes make their management difficult. The present chapter mainly has three major sections: (1) mosquito, viruses, interrelationships and disease treatment, (2) vector management and (3) novel approaches for vector management and disease transmission inhibition. The first aspect includes the relationship of mosquitoes with arboviruses, vector competency and the treatments of arbovirus diseases involving vaccine developments with future aspects. The next section deals with mosquito biology, surveillance and control strategies for eggs, larvae, pupae and adults stages using the conventional and advanced methods to develop an integrated approach for effective vector management. This section also elaborates the environmental management, biological and insecticidal controls of different stages of mosquitoes. The last segment of the chapter discusses the efficacy and applicability of novel technologies for multiple stages of Aedes mosquito control which includes autodissemination technology, Wolbachia-based cytoplasmic incompatibility, sterile insect techniques and the release of insects carrying a dominant lethal gene (RIDL) that can be a potential tool in the future to restrict disease transmission. The human behaviour alteration to avoid mosquito biting and methods applicable for personal protection has been discussed in the chapter. Based on the discussion, the prevention of the transmission of dengue, chikungunya and Zika is the only cure in the present scenario.

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 Kruppel-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.

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.

Superparasitism is a common phenomenon in mosquito-parasitic mermithid nematodes. Multiple nematodes are needed in a single host to produce males. Host selection behavior and intraspecific competition among Romanomermis iyengari and Strelkovimermis spiculatus were investigated against their host, Culex pipiens pipiens in laboratory experiments. In a choice assay between previously infected and uninfected host larvae, infectious preparasites of both nematode species could distinguish not only between infected and uninfected hosts, but even between different parasite loads in showing a strong preference for uninfected hosts or hosts with a low parasite load. Host heart rate declined briefly immediately after parasitism. Superparasitism resulted in increased parasite mortality. Scramble competition within mosquito larvae for limited host nutrients, coupled with a skewed sex ratio favoring males, is assumed to lead to parasite population decline and subsequently toward host-parasite population equilibrium. The ability of mermithid preparasites to accurately assess parasite load likely plays an important role in host population dynamics and regulation.

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.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 LAutodissemination 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.

a b s t r a c t Suppression of Aedes albopictus populations is a substantial challenge for mosquito control programs globally because juveniles of this species are found in numerous kinds of domestic artificial containers that are difficult to detect, access, and eliminate. We conducted a multi-year assessment of the effect of different interventions to control Ae. albopictus near the northernmost geographic boundary of the species in temperate North America and deployed an array of BG-Sentinel traps for adult surveillance. Here we present the results of a comparative examination of adult sex ratios in urban and suburban areas, shifts in sex ratios after control interventions, and a discussion of the critical drivers of population dynamics of Ae. albopictus in our area. We collected significantly more male mosquitoes in urban as compared to suburban areas in June through September, but not in May (p < 0.001). The higher number of male mosquitoes in urban areas could be attributed to a higher number of larval habitats within a closer proximity of the surveillance traps and the lower flight dispersal of males. Following application of adulticides in urban areas, Ae. albopictus male populations were reduced by 88% on average, which was higher than the 69% reduction in female populations. The higher reduction of male mosquitoes could be attributed to the smaller body mass of the males and their higher susceptibility to adulticides. The results of this study are directly relevant to the development of suitable control strategies that depend on manipulation of males, such as the sterile insect technique. The results could also be used to refine mosquito abatement by providing more accurate methods to determine the need and timing of vector

a b s t r a c t Autodissemination of insecticides is a novel strategy for mosquito management. We tested if contami- nated Aedes albopictus (Skuse) mosquitoes from a small area treated with commercial formulations (79 gm 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 effi- cacy 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 autodissem- ination in four compass directions out to 200-400 m 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 105 ha) using backpack sprayer in 2010 and in strips (24.8% of 94 ha) 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.

Mating aggregations in the mosquito parasitic nematode, Strelkovimermis spiculatus, were investigated in the laboratory. Female postparasites, through their attraction of males and, remarkably, other females, drive the formation of mating clusters. Clusters may grow in size by merging with other individual or clusters. Female molting to the adult stage and reproductive success are enhanced in larger clusters. Male mating behavior is initiated when the female begins to molt to the adult stage by shedding dual juvenile cuticles posteriorly. Males coil their tail around the adult cuticle, migrating progressively along the female in intimate synchrony with the molting cuticle until the vulva is exposed and mating can occur. The first arriving male is assured of access to a virgin female, as his intermediate location between the vulva and subsequently arriving males blocks these competitors. Males deposit an adhesive gelatinous copulatory plug into and over the vulva before departing the female. Fecundity was greater in larger mating clusters, but this was a function of a greater rate of molting which is a prerequisite for mating. Males compete for virgin females by emerging and molting to the adult stage earlier than females. Mating aggregations have previously only been examined in snakes, but these studies have tended to be observational as snakes offer a challenging system for study. The relatively easy to culture and manipulate mermithid system may offer a model for experimental studies of male-male competition, protandry, copulatory plugs and female choice in mating clusters.

BACKGROUND:TheAsiantigermosquito,Aedesalbopictus(Skuse),transmitsimportantarboviraldiseasesanddisplacesnative 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 (31m 3 ) bioassay a single station consistently achieved 100% Ae.albopictus pupal mortality against 10 oviposition containers. In a greenhouse (200m 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. c � 2013 Society of Chemical Industry

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.

The Asian tiger mosquito, Aedes albopictus (Skuse), is an invasive species and a major pest problem in urban and suburban locales in New Jersey. To assess its potential role as an arbovirus vector, we sampled Ae. albopictus from two New Jersey counties over a 3-yr period and estimated the prevalence of virus infection by Vero cell culture and reverse transcription-polymerase chain reaction assays. Three virus isolates were obtained from 34,567 field-collected Ae. albopictus, and all were identified as Cache Valley virus by molecular methods. Ae. albopictus (N = 3,138), collected in Mercer County from late July through early September 2011, also were retested for West Nile virus (WNV) by reverse transcription-polymerase chain reaction, and all were negative. These results corroborate previous findings showing that Ae. albopictus may occasionally acquire Cache Valley virus, a deer-associated arbovirus, in nature. In contrast, we did not detect WNV infection in Ae. albopictus despite concurrent WNV amplification in this region.

a b s t r a c t 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 1.0 ppm concentrations. Egg hatching inhibition was dose dependent, the highest being at 1.0 ppm 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 (5.1 h) as compared to Ae. aegypti (4.9 h), Ae. albopictus (3.9 h) or Cx. pipiens (1.7 h) 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.

Lipid metabolism is coordinately regulated through signaling networks that integrate biochemical pathways of fat assimilation, mobilization and utilization. Excessive diversion of fat for storage is a key risk factor for many fat-related human diseases. Dietary lipids are absorbed from the intestines and transported to various organs and tissues to provide energy and maintain lipid homeostasis. In humans, disparity between triglycerides (TG) synthesis and removal, via mitochondrial β-oxidation and VLDL (very low density lipoprotein) secretion, causes excessive TG accumulation in the liver. The mutation in Caenorhabditis elegans KLF-3 leads to high TG accumulation in the worm's intestine. Our previous data suggested that klf-3 regulates lipid metabolism by promoting fatty acid β-oxidation. Depletion of cholesterol in the diet has no effect on fat deposition in klf-3 (ok1975) mutants. Addition of vitamin D in the diet, however, increases fat levels in klf-3 worms. This suggests that excess vitamin D may be lowering the rate of fatty acid β-oxidation, with the eventual increase in fat accumulation. We also demonstrate that mutation in klf-3 reduces expression of C. elegans dsc-4 and/or vit genes, the orthologs of mammalian microsomal triglyceride transfer protein and apolipoprotein B, respectively. Both microsomal triglyceride transfer protein and apolipoprotein B are essential for mammalian lipoprotein assembly and transport, and mutation in both dsc-4 (qm182) and vit-5 (ok3239) results in high fat accumulation in worm intestine. Genetic interactions between klf-3 and dsc-4, as well as vit-5 genes, suggest that klf-3 may have an important role in regulating lipid assembly and secretion.

Background Aedes (Stegomyia) albopictus (Skuse) is an important disease vector and biting nuisance. During the 2009 active season, six ∼1000-parcel sites were studied, three in urban and three in suburban areas of New Jersey, United States, to examine the efficacy of standard integrated urban mosquito control strategies applied area wide. Active source reduction, larviciding, adulticiding and public education (source reduction through education) were implemented in one site in each county, an education-only approach was developed in a second site and a third site was used as an untreated experimental control. Populations were surveyed weekly with BG-Sentinel traps and ovitraps. Results A substantial reduction in Ae. albopictus populations was achieved in urban sites, but only modest reductions in suburban sites. Education alone achieved significant reductions in urban adult Ae. albopictus. Egg catches echoed adult catches only in suburban sites. Conclusions There are significant socioeconomic and climatic differences between urban and suburban sites that impact upon Ae. albopictus populations and the efficacy of the control methods tested. An integrated pest management approach can affect abundances, but labor-intensive, costly source reduction was not enough to maintain Ae. albopictus counts below a nuisance threshold. Nighttime adult population suppression using truck-mounted adulticides can be effective. Area-wide cost-effective strategies are necessary. Published 2013. This article is a U.S. Government work and is in the public domain in the USA

Background Dietary salts are important factors in metabolic disorders. They are vital components of enzymes, vitamins, hormones, and signal transduction that act synergistically to regulate lipid metabolism. Our previous studies have identified that Krüppel-like factor −3 (KLF-3) is an essential regulator of lipid metabolism. However, it is not known if KLF-2 also regulates lipid metabolism and whether KLF-2 and −3 mediate the effects of dietary salts on lipid metabolism. Methods In this study, we used klf mutants [homozygous klf-2 (ok1043) V and klf-3 (ok1975) II mutants] to investigate the role of dietary salts in lipid metabolism. All gene expression was quantified by qRT-PCR. Localization of KLF-2 was analyzed by the expression of klf-2::gfp (in pPD95.75 vector) using a fluorescent microscope. Fat storage was measured by Oil Red O staining. Results Klf-2 was identified to express in the intestine during all stages of Caenorhabditis elegans development with peak expression at L3 stage. Mutation of klf-2 increased fat accumulation. Under regular growth media free of Ca2+, the expression of both klf-2 and −3 was inhibited slightly; further their expression reduced significantly in WT worms fed on 10X Ca2+ diet. When klf-3 was mutated, the expression of klf-2 increased under 10X Ca2+ diet; but when klf-2 was mutated, the expression of klf-3 was not altered under 10X Ca2+ diet. Overall, Mg2+ and K+ were less effective on the gene expression of klfs. KLF target gene Ce-C/EBP-2 showed elevated expression in WT and klf-3 (ok1975) worms with changed Ca2+ concentrations but not in klf-2 (ok1043) worms. However, high Ca+2 diet exhibited inhibitory effect on Ce-SREBP expression in WT worms. Conclusion Dietary Ca2+ is most effective on fat storage and klf-2 expression, wherein high Ca2+ diet decreased klf-2 expression and reduced fat buildup. Mechanistic study identified Ce-C/EBP (C48E7.3; lpd-2) and Ce-SREBP (Y47D3B.7; lpd-1) as the target genes of klf-2 and/or klf-3 to mediate lipid metabolism. This study identifies a new function of klf-2 in inhibiting fat buildup and reveals the interplay between dietary salts and klf-2 and klf-3 in lipid metabolism.

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

Adult control of Aedes albopictus via ultra-low volume is difficult because this species occurs primarily in peridomestic habitats where obstacles such as buildings and vegetation can disrupt spray plumes and droplet dispersion. We determined droplet penetration and characterization of a pyrethroid adulticide applied from the ground at mid (46.77 ml/ha) and maximum (93.53 ml/ha) label rates within cryptic habitats of urban and suburban environments. Droplets were collected from all habitats, with no significant differences detected between locations within the same application rate or collection method. No differences were detected in droplet densities (drops per mm2) between rates within urban environments, but more droplets were collected in urban (149.93 ± 11.07 SE) than suburban sites (114.37 ± 11.32) at the maximum label rate (P = 0.003). The excellent penetration of aerosols into cryptic habitats of an urban site was likely due to the shorter spray paths afforded by our network of roads and alleys. Mid label rates displayed similar droplet density values as maximum label rates in urban areas, indicating that lower rates may be used effectively to reduce costs, lessen non-target effects, and increase environmental stewardship. Advances in formulations and technology are driving changes in adulticide applications, leading to use of the minimum effective dose for maximum efficacy, precision, and accountability.

Pyriproxyfen, a juvenile hormone analogue, diflubenzuron, a chitin synthesis inhibitor, and azadirachtin, an ecdysone agonist, are three insect growth regulators (IGRs) considered as selective and effective insecticides for mosquitoes. Romanomermis iyengari (Welch) is a mosquito-parasitic mermithid that can provide biological control against many medically important mosquito species. The compatibility of these two control tactics was tested by evaluating the sublethal effects of exposure to IGR on nematode developmental stages (preparasitic, parasitic, and preparasitic + parasitic) using Culex pipiens larvae as the host. Sublethal concentrations of IGRs were 90 % emergence inhibition of host mosquito. Preparasitic exposure to pyriproxyfen, azadirachtin, and diflurbenzuron had no effect on infectivity, parasite load, sex ratio, or male size but reduced nematode female length and increased male sex ratio at one parasite/larva. When IGRs treatments were made against the parasitic and preparasitic + parasitic stages, pyriproxyfen and azadirachtin reduced R. iyengari infectivity, parasite load, and male nematode length, whereas pyriproxyfen exposure increased male sex ratio and reduced the female R. iyengari length. Thus, IGRs have significant negative impacts on different stages of mosquito mermithid that can destabilize the balance of host-parasite population interaction. Therefore, IGRs should be used with caution in mosquito habitats where these parasites have established.