Behavior manipulation of mosquitoes by a mermithid nematode.

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.

The role of copulatory plugs in the mosquito-parasitic nematode Strelkovimermis spiculatus.

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.

Superparasitism and Population Regulation of the Mosquito-Parasitic Mermithid Nematodes Romanomermis iyengari and Strelkovimermis spiculatus.

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.

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.

Long-term Effects of Carbohydrate Availability on Mating Success of Newly Eclosed Aedes albopictus (Diptera: Culicidae) Males.

Sugar availability varies greatly in nature, and determining how this affects male mosquito fitness is essential for understanding population dynamics. We allowed male Aedes albopictus (Skuse) carbohydrate access for increasing intervals of time immediately after eclosion and we evaluated their fitness by comparing mortality, mating success, and sperm transfer. We compared individual male Ae. albopictus, which were offered water or 20% sucrose solution for 24, 48, or 72 h. As predicted, there were significant increases in fitness for each additional day of sucrose access. Following sugar exposure, we allowed males daily access to three virgin females. We assessed mating success through observation of spermatozoa in the female spermathecae. When individuals of the same age were compared, males with sugar access exhibited significantly greater mating success than water-treated males in all treatments. The total number of spermathecae filled by males with sugar access in the 48- and 72-h treatments was also significantly greater on some days; these were 3-5 d posteclosion in the 48-h treatment and 5-6 d posteclosion in the 72-h treatment. We conclude that extended sugar access at eclosion is important for maximizing fitness in male Ae. albopictus and should be applicable to sterile male release efforts, especially when laboratory-reared males suffered from other disadvantages. We recommend retaining adult males for 3 d posteclosion prior to release to improve their mating success in male release initiatives.

Mating clusters in the mosquito parasitic nematode, Strelkovimermis spiculatus.

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.

Vertical Oviposition Preferences of the Asian Tiger Mosquito, Aedes albopictus, In Temperate North America.

As a hyperaggressive mosquito that is also a public health threat, Aedes albopictus (Skuse), the Asian tiger mosquito, is a major priority for control efforts. We examine one aspect of Ae. albopictus biology: oviposition height. Field-based research in an urban habitat was conducted to determine if a height preference exists for this species. Larval and egg counts showed a significant preference for oviposition at ground level (0 m) compared to heights of 1, 2, 3, or 4 m (P < 0.01). An experiment conducted under semi-field conditions supported our conclusion of oviposition preference at ground level (P < 0.001), and further defines the search image needed by mosquito control personnel when dealing with this invasive species.

Taming a tiger in the city: comparison of motorized backpack applications and source reduction against the Asian tiger mosquito, Aedes albopictus.

We evaluated 2 strategies to manage Aedes albopictus: 1) motorized backpack applications and 2) source reduction (coupled with hand-applied applications of larvicide). Backpack applications used a water-dispersible granular formulation (VectoBac WDG) of Bacillus thuringiensis var. israelensis (Bti), whereas source reduction used granular formulations of the insect growth regulator methoprene (Altosid) combined with a monomolecular film surfactant (Agnique). Six subplots (total 8.02 ha) were selected for backpack applications, source reduction, and control groups. The experiments were blind with applications conducted randomly and independently. Efficacy was determined through placement of bioassay cups with larvae within experimental plots 1 day before treatment. Backpack applications resulted in 76% (+/- 8.2% SE) and source reduction resulted in 92% (+/- 4.1% SE) larval mortality. Backpack applications required 50 times less labor than source reduction (0.25 versus 0.005 ha/h). The cost of backpack applications, including labor, was $159.88/ha, compared with $659.65/ha for source reduction. Although overall efficacy was slightly lower, motorized backpack applications of Bti were more efficient and cost-effective than source reduction methods to control Ae. albopictus in urban settings at the community level.

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.

Isolations of Cache Valley virus from Aedes albopictus (Diptera: Culicidae) in New Jersey and evaluation of its role as a regional arbovirus vector.

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.

Effects of insect growth regulators on the mosquito-parasitic nematode Romanomermis iyengari.

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.

Child outdoor physical activity is reduced by prevalence of the Asian tiger mosquito, Aedes albopictus.

We tested the hypothesis that day-biting mosquitoes contribute to child obesity by reducing opportunities for summer outdoor play. The influence of Asian tiger mosquito (Aedes albopictus) prevalence on child outdoor physical activity was compared in 2 matched urban communities, one treated for mosquito abatement and one untreated. More time was spent outdoors by children where abatement took place.

Assessment of a direct application of two Bacillus thuringiensis israelensis formulations for immediate and residual control of Aedes albopictus.

The bacterial agent Bacillus thuringiensis israelensis (Bti) is a highly effective larvicide against various medically important mosquito and black fly vector species. Recent formulations of this powerful larvicidal tool have been evaluated for their field efficacy in integrated mosquito management programs. Laboratory and controlled-condition trials have indicated long periods of residual efficacy, whereas field persistence is often much lower in duration. We investigated the residual persistence of high doses of 2 formulations of Bti, a water-dispersible granule (VectoBac WDG; 16 mg/liter) and an extruded pellet (VBC-60066; 80 mg/liter), for the management of natural larval populations of Aedes albopictus. Laboratory tests demonstrated 100% (WDG) and > or = 99.7% (VBC) average mortality across all treatments over 180 days. Field tests exhibited 100% efficacy (WDG and VBC) for 3 wk against Ae. albopictus and other coinhabiting mosquito species, with some residual efficacy lasting for > 4 wk. These results are discussed in relation to current field control of domestic Aedes vectors of public health significance.

Host Penetration and Emergence Patterns of the Mosquito-Parasitic Mermithids Romanomermis iyengari and Strelkovimermis spiculatus (Nematoda: Mermithidae).

Romanomermis iyengari and Strelkovimermis spiculatus are mermithid nematodes that parasitize mosquito larvae. We describe host penetration and emergence patterns of Romanomermis iyengari and Strelkovimermis spiculatus in laboratory exposures against Culex pipiens pipiens larvae. The mermithid species differed in host penetration behavior, with R. iyengari juveniles attaching to the host integument before assuming a rigid penetration posture at the lateral thorax (66.7%) or abdominal segments V to VIII (33.3%). Strelkovimermis spiculatus attached first to a host hair in a coiled posture that provided a stable base for penetration, usually through the lateral thorax (83.3%). Superparasitism was reduced by discriminating against previously infected hosts, but R. iyengari's ability to avoid superparasitism declined at a higher inoculum rate. Host emergence was signaled by robust nematode movements that induced aberrant host swimming. Postparasites of R. iyengari usually emerged from the lateral prothorax (93.2%), whereas S. spiculatus emergence was peri-anal. In superparasitized hosts, emergence was initiated by males in R. iyengari and females in S. spiculatus; emergence was otherwise nearly synchronous. Protandry was observed in R. iyengari. The ability of S. spiculatus to sustain an optimal sex ratio suggested superior self-regulation. Mermithid penetration and emergence behaviors and sites may be supplementary clues for identification. Species differences could be useful in developing production and release strategies.

Partners in diabetes epidemic: A global perspective.

There is a recent increase in the worldwide prevalence of both obesity and diabetes. In this review we assessed insulin signaling, genetics, environment, lipid metabolism dysfunction and mitochondria as the major determinants in diabetes and to identify the potential mechanism of gut microbiota in diabetes diseases. We searched relevant articles, which have key information from laboratory experiments, epidemiological evidence, clinical trials, experimental models, meta-analysis and review articles, in PubMed, MEDLINE, EMBASE, Google scholars and Cochrane Controlled Trial Database. We selected 144 full-length articles that met our inclusion and exclusion criteria for complete assessment. We have briefly discussed these associations, challenges, and the need for further research to manage and treat diabetes more efficiently. Diabetes involves the complex network of physiological dysfunction that can be attributed to insulin signaling, genetics, environment, obesity, mitochondria and stress. In recent years, there are intriguing findings regarding gut microbiome as the important regulator of diabetes. Valid approaches are necessary for speeding medical advances but we should find a solution sooner given the burden of the metabolic disorder - What we need is a collaborative venture that may involve laboratories both in academia and industries for the scientific progress and its application for the diabetes control.

KLF regulation of insulin pathway genes.

Alteration in lipid metabolism can result in fat accumulation in adipose tissues, which may lead to two most important human diseases, obesity and diabetes. A shift in lipid metabolism deregulates signaling pathways which regulates obesity and/or diabetes. In this study, we examined the components of insulin/ TGF-ß pathways and their genetic interaction with Krüppel-like transcription factors (KLFs). Their role in energy homeostasis were discussed. We separately created klf/daf genes double mutants by carrying out klfs RNAi on daf-2 (e1391), daf-4 (e1364), daf-7 (e1372); dpy-1 (e1), daf-14 (m77), daf-16 (mgDf50) mutants. And then conducted Oil O Red staining to assay the klf/daf RNAi worms for fat deposits and examine genetic interaction between klfs and daf genes. The results showed that worms bearing klf-1, 2, or 3 and daf-2, or daf-4 mutations deposit large, but similar fat levels as individual mutants. The results suggested that they target the same molecular pathway of fat storage. klf-1, 2 or 3 RNAi /daf-7 worms showed higher fat deposits in klf-1, 2, or 3 RNAi/daf-7 worms than klf-1, 2, or 3 RNAi or daf-7 mutants alone, which showed a functional interaction between klfs and daf-7 in perhaps TGF-ß-like pathway. Altogether our study suggests a direct role of klfs in insulin signaling pathway.

Strain specific differences in intraspecific competition in Aedes albopictus (Diptera: Culicidae).

Malathion is an organophosphate insecticide that is used for the control of adult mosquitoes and agricultural pests. Recent studies have shown that malathion affects competition among mosquitoes in the larval stage. Individuals from laboratory colonies are often used in experiments but it is not known whether there is a difference between laboratory and field strains in their response to competition and malathion. Intraspecific larval competition in the presence of malathion (0.11 mg/liter) was compared between laboratory and field strains of Aedes albopictus (Skuse), a native of Asia that has established in the United States. There was no difference in the responses of the two strains to the presence of malathion. The fitness (finite growth rate) of the field strain decreased at the highest larval density tested but there was no difference in fitness across densities for the laboratory strain. This finding suggests that laboratory rearing could reduce sensitivity to crowding.

Larval mosquito habitat utilization and community dynamics of Aedes albopictus and Aedes japonicus (Diptera: Culicidae).

Aedes albopictus (Skuse) and Ae. japonicus (Theobald) are important container-inhabiting mosquitoes that transmit disease agents, outcompete native species, and continue to expand their range in the United States. Both species deposit eggs in natural and artificial containers and thrive in peridomestic environments. The goal of our study was to examine the types and characteristics of containers that are most productive for these species in the northeastern United States. In total, 306 containers were sampled in urban, suburban, and rural areas of New Jersey. Multiple biotic and abiotic factors were recorded in an attempt to identify variables associated with the productivity of each species. Based on pupal abundance and density of container types, results showed that tires, trash cans, and planter dishes were the most important containers for Ae. albopictus, while planter dishes were the most important containers for Ae. japonicus. Container color (black and gray), material (rubber), and type (tires) were correlated with species presence for Ae. albopictus and Ae. japonicus. These factors may play a role in the selection of oviposition sites by female mosquitoes or in the survival of their progeny. Differences in species composition and abundance were detected between areas classified as urban, suburban, and rural. In urban and suburban areas, Ae. albopictus was more abundant in container habitats than Ae. japonicus; however, Ae. japonicus was more abundant in rural areas, and when water temperatures were below 14 degrees C. Our results suggest many variables can influence the presence of Ae. albopictus and Ae. japonicus in container habitats in northeastern United States.

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.

Willingness-to-pay for an area-wide integrated pest management program to control the Asian tiger mosquito in New Jersey.

Using contingent valuation we estimated the perceived value of an area-wide integrated pest management program for the Asian tiger mosquito, Aedes albopictus, implemented in Monmouth and Mercer counties, NJ. We estimated residents' maximum willingness-to-pay and perceived monetary benefits (willingness-to-pay excluding residents who protested all types of payments) and payment modality through a telephone survey of 51 randomly selected households. The mean (+/- SE) perceived monetary benefits for an enhanced mosquito abatement program was $9.54 +/- 2.90 per capita per year. Most respondents would have been willing to pay through taxes (35%) or charitable donations (6%) starting then, or through one of these approaches in the future (43%), whereas 16% were completely unwilling to pay any additional costs whatsoever. We projected that the perceived monetary benefits to the counties' 1.01 million residents for an enhanced mosquito control program would be $9.61 million annually. Thus, collectively residents perceived monetary benefits of 3.67 times the combined 2008 annual operating costs of the counties' existing mosquito control programs of $2.61 million.