Detection of Babesia microti and Borrelia burgdorferi in host-seeking Ixodes scapularis (Acari: Ixodidae) in Monmouth County, New Jersey.

The etiological agents that cause human babesiosis (Babesia microti) and Lyme disease (Borrelia burgdorferi) share a common tick vector (Ixodes scapularis Say) and rodent reservoir (Peromyscus leucopus), but because the geographical distribution of babesiosis is more restricted than Lyme disease, it was not considered a nationally notifiable disease until 2011. Although recent studies have shown dramatic increases in the number of cases of babesiosis and expansion of its range, little is known about infection and coinfection prevalence of these pathogens in the primary tick vector. Of the 478 I. scapularis nymphs collected within six Monmouth County, NJ, municipalities between 2004 and 2006, 4.0 and 10.0% were infected with B. microti and B. burgdorferi, respectively, while 2.9% were coinfected. Analysis of the 610 I. scapularis adults collected during the same period yielded an infection prevalence of 8.2% for B. microti and 45.2% for B. burgdorferi, while 6.2% were coinfected. The potential public health importance of these findings is discussed.

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.

Experiences with the large-scale operation of the Biogents Sentinel trap.

We obtained 160 Biogents Sentinel traps (BGS-traps) to monitor adult mosquito populations for the Area-wide Pest Management Program for the Asian tiger mosquito (Aedes albopictus) in New Jersey. We deployed between 90 and 110 BGS-traps weekly from May through October of 2008, 2009, 2010. and 2011. Here we detail our experience: challenges with acquisition, defects in construction, as well as actions taken to correct problems we found and preempt them in the future. Further, we describe the impact of these problems on our research and provide a cost analysis of repairs.

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.

A model to predict evaporation rates in habitats used by container-dwelling mosquitoes.

Container-dwelling mosquitoes use a wide variety of container habitats. The bottle cap is often cited as the smallest container habitat used by container species. When containers are small, the habitat conditions can greatly affect evaporation rates that in turn can affect the species dynamics within the container. An evaporation rate model was adapted to predict evaporation rates in mosquito container habitats. In both the laboratory and field, our model was able to predict actual evaporation rates. Examples of how the model may be applied are provided by examining the likelihood of Aedes albopictus (Skuse), Aedes aegypti (L.), and Culex pipiens pipiens (L.) completing their development within small-volume containers under typical environmental conditions and a range of temperatures. Our model suggests that under minimal direct sunlight exposure, both Ae. aegypti and Ae. albopictus could develop within a bottle cap before complete evaporation. Our model shows that under the environmental conditions when a plastic field container was sampled, neither Ae. albopictus or Cx. p. pipiens could complete development in that particular container before the water evaporated. Although rainfall could replenish the habitat, the effects of evaporation would increase larval density, which could in turn further decrease developmental rates.

Geographical distribution and prevalence of selected Borrelia, Ehrlichia, and Rickettsia infections in Amblyomma americanum (Acari: Ixodidae) in New Jersey.

We used published accounts to describe the known statewide distribution of the lone star tick, Amblyomma americanum, in New Jersey and field surveys to characterize the geographical range of A. americanum and selected A. americanum-transmitted pathogens in Monmouth County, the hypothesized northern limit of the species distribution. Ticks were collected using standardized methods from 50 widely dispersed public access areas within 18 municipalities to produce estimates of relative abundance among sites. Collected A. americanum adults were stored at -80 degrees C until processed for DNA extraction. Individual ticks were subjected to polymerase chain reaction analyses to detect the presence of Ehrlichia chaffeensis, E. ewingii, Rickettsia amblyommii, and Borrelia lonestari. The range of A. americanum was generally limited to the southern half of New Jersey. Within Monmouth County, we collected A. americanum from 9 of 18 municipalities (50%) and 24 (48%) of the surveyed properties. We found at least 1 pathogen at 17 (70.8%) sites located within 6 of 9 municipalities, while all 4 target pathogens were detected in 5 of those 6 (83.3%) municipalities. The geographical distribution of A. americanum and its associated pathogens appeared to be restricted to the southern portion of the county. Possible factors governing the distribution are discussed.

A droplet collection device and support system for ultra-low-volume adulticide trials.

A supporting stand to suspend rotating impactors and mosquito cages is a requirement for field tests during pesticide efficacy trials. We present schematics for a collection device and associated support system for sampling droplets of ultra-low-volume (ULV) sprays during mosquito adulticide applications. This system offers the advantages of cost efficiency, increased ease of deployment, off-season storage, visibility, stability, and ULV collection efficacy. Use of this system ensures that droplet collection and meteorological equipment is at appropriate and consistent heights between trials. The 2 arms of the support allow for placement of multiple cages, light-emitting or reflecting devices, and wind-indicating ribbons to be attached to the station. The support described makes possible deployment of stations over a wider variety of terrains, increasing the extent of field trials. Presentation of the simple design and fabrication of the rotating collection device (impactor), T-station, and its support is provided.

Field efficacy of BG-Sentinel and industry-standard traps for Aedes albopictus (Diptera: Culicidae) and West Nile virus surveillance.

Standard surveillance traps in North America for adult Aedes albopictus (Skuse) (Diptera: Culicidae), an invasive mosquito with public health implications, are currently ineffective. We compared the efficacy of the BG-Sentinel trap (BGS) with and without lures (BG-lure, octenol, and CO2), the Centers for Disease Control and Prevention light trap (CDC) with and without lures, and the gravid trap (GT) for Ae. albopictus collection in two urban sites in New Jersey. The BGS with or without lures collected more Ae. albopictus compared with other trap configurations and was more specific for Ae. albopictus. In Camden County, the BGS with lures collected three times more Ae. albopictus than the CDC (with CO2 only) and five times more than the GT. In Mercer County, BGS with lures collected the most mosquitoes, with 3 times more Ae. albopictus than the CDC with all lures and 50 times more than the GT. The BGS collected more male Ae. albopictus than other traps in both counties, providing further population monitoring. The GT and BGS provided a relative measure of the enzootic activity of West Nile virus in Culex spp. and the potential epidemic activity of WNV in Ae. albopictus. The BGS provides effective chemical and visual cues for host-seeking Ae. albopictus and should be used as a part of existing surveillance programs and new initiatives targeting this mosquito.

Sentinel chicken coop modification for canopy-level arbovirus disease surveillance.

A pulley-block system for elevating sentinel chicken coops was designed to conduct mosquito-borne virus surveillance in the forest canopy. This modified design allows traditional captive sentinel bird cages to be hoisted easily to the desired height and lowered at regular intervals for routine maintenance and sampling of the flocks. The design specifications of this canopy system are modifications that can easily be incorporated to existing rabbit hutches used in ground-level captive sentinel systems.

Ability of 4-poster passive topical treatment devices for deer to sustain low population levels of Ixodes scapularis (Acari: Ixodidae) after integrated tick management in a residential landscape.

In a recent study, the combined use of 4-Posters and Maxforce TMS bait boxes along with a barrier application of deltamethrin resulted in accelerated control of Ixodes scapularis Say by sequentially attacking each postembryonic life stage. We report the results of a follow-up study to test the ability of 4-Posters used alone to sustain the high levels of control achieved through the integrated tick management (ITM) approach after withdrawal of the bait boxes. In the first year after withdrawal, we observed declines in the level of control of larvae on small mammals, as well as of numbers of host-seeking larvae in the treatment area. There was no difference in the level of control of host-seeking adults in the treatment area after 2 yr. Within 2 yr, we observed a decline in control of subadult ticks infesting small mammals, but continued to see significant control of both host-seeking nymphs (85.9%) and larvae (89.0%) in the treatment area. The inconsistency that we observed between the apparent ability of 4-Posters to sustain high levels of control of host-seeking ticks, although having less effect on tick burdens on small mammal hosts, may be explained by the host-seeking ecology of immature I. scapularis.

Suppression of Ixodes scapularis (Acari: Ixodidae) following annual habitat-targeted acaricide applications against fall populations of adults.

Spring acaricide applications directed against nymphal Ixodes scapularis have been shown to be effective, but are perceived by the public as having significant adverse environmental impacts, particularly against nontarget organisms. Targeting the adult stage of I. scapularis in the fall would hypothetically result in indirect control of subsequent subadult stages while avoiding other arthropods that are typically inactive during this period. We demonstrate that single fall applications of deltamethrin for 3 consecutive years immediately reduced fall questing adults, while also rapidly reducing the abundance of all postembryonic stages. Deltamethrin applied to the shrub-layer vegetation resulted in levels of control between 97.1% and 100% at 7 days postapplication. Repeated applications against the reproductive stage of I. scapularis progressively reduced the abundance of larvae and nymphs in treated plots, reaching 91.4% and 100% by the conclusion of the study.

A historical snapshot of Ixodes scapularis-borne pathogens in New Jersey ticks reflects a changing disease landscape.

Historical specimens, when available, can provide new insight into the distribution and evolution of pathogens that may not be discernible from more recent samples. We used ticks collected from hunter-killed white-tailed deer in New Jersey in 2002 to examine the prevalence and distribution of four pathogens transmitted by Ixodes scapularis, the blacklegged tick. Infection with Borrelia burgdorferi sensu stricto, Babesia microti, and Anaplasma phagocytophilum (the agents of Lyme disease, human babesiosis, and human granulocytic anaplasmosis, respectively) was highest in the Coastal Plain and lowest in the northwestern Skylands region. These patterns correspond well with the historically observed northward expansion of I. scapularis within New Jersey and the comparatively recent increase in human cases of these pathogens in the northern part of the state. Additionally, we provide evidence that Borrelia miyamotoi, a relatively new emerging pathogen and agent of relapsing fever, was widespread (though not common) throughout the state in 2002. Our findings highlight the need for enhanced awareness of tick-borne diseases other than Lyme and implementation of large-scale tick surveillance in endemic regions.

Integrated use of 4-Poster passive topical treatment devices for deer, targeted acaricide applications, and Maxforce TMS bait boxes to rapidly suppress populations of Ixodes scapularis (Acari: Ixodidae) in a residential landscape.

In fall 2003, we began testing an integrated control strategy to rapidly achieve and sustain reduced numbers of Ixodes scapularis Say (Acari: Ixodidae) in a residential area. We combined two host-targeted technologies in conjunction with single, barrier acaricide applications to sequentially attack each postembryonic life stage of the tick. Granular deltamethrin applied to the lawn-forest interface of participant properties resulted in 100% control of host-seeking nymphs. Nymphal and larval tick burdens on targeted small mammal hosts at treated properties were reduced by 92.7 and 95.4%, respectively, after the first year (2004) of combined interventions. Over the same period, populations of host-seeking nymphs, larvae, and adults were reduced by 58.5, 24.8, and 77.8%, respectively. After interventions in 2005, tick burdens on small mammals were maintained at similar levels, whereas control of host-seeking nymphs, larvae, and adults increased to 94.3, 90.6, and 87.3%, respectively. Prospects for widespread use of these technologies to protect the public's health are discussed.

Effect of Rickettsia felis Strain Variation on Infection, Transmission, and Fitness in the Cat Flea (Siphonaptera: Pulicidae).

Rickettsia felis is a human pathogen transmitted by the cat flea, Ctenocephalides felis (Bouché) (str. LSU), as well as an obligate symbiont of the parthenogenic booklouse Liposcelis bostrychophila (Badonnel) (str. LSU-Lb). The influence of genetic variability in these two strains of R. felis on host specialization and fitness and possible resulting differences on infection and transmission kinetics in C. felis is unknown. Utilizing an artificial host system, cat fleas were exposed to a R. felis str. LSU-Lb-infected bloodmeal and monitored for infection at 7-d intervals for 28 d. Quantitative real-time PCR was used to determine rickettsial load and infection density in newly exposed cat fleas, and transmission frequency between cat fleas. The effect of persistent R. felis infection on cat flea F1 progeny was also assessed. At 7 d postexposure 76.7% of the cat fleas successfully acquired R. felis str. LSU-Lb. In R. felis str. LSU-Lb-exposed cat fleas, the mean infection load (6.15 × 106), infection density (0.76), and infection prevalence (91/114) were significantly greater than R. felis str. LSU infection load (3.09 × 106), infection density (0.68), and infection prevalence (76/113). A persistent R. felis str. LSU-Lb infection was detected for 28 d in adult cat fleas but neither female:male ratio distortion nor vertical transmission was observed in F1 progeny. While infection kinetics differed, with higher intensity associated with R. felis str. LSU-Lb, no distinct phenotype was observed in the F1 progeny.

Relative abundance and prevalence of selected Borrelia infections in Ixodes scapularis and Amblyomma americanum (Acari: Ixodidae) from publicly owned lands in Monmouth County, New Jersey.

To evaluate their potential importance in the transmission of ixodid tick-borne borrelioses in Monmouth County, NJ, we collected host-seeking Ixodes scapularis Say and Amblyomma americanum (L.) (Acari: Ixodidae) adults and nymphs to determine relative encounter frequencies and the infection prevalence of selected Borrelia spp. in their respective tick vectors. We also reviewed records of all ticks submitted for identification by the public in Monmouth County during 2001-2005. Relative abundance of the two species varied markedly among sites. Adult encounter frequencies for the two species were similar; however, A. americanum nymphs were encountered 3 times more frequently than I. scapularis nymphs. Of 435 ticks submitted by the public, 50.1 and 38.9% were I. scapularis and A. americanum, respectively. However, during May through August, the peak Lyme disease transmission season in New Jersey, significantly more submitted ticks were A. americanum (55.9%), compared with I. scapularis (34.1%). Polymerase chain reaction analysis of 94 1. scapularis and 103 A. americanum adults yielded infection prevalences of 31.9% for B. burgdorferi and 5.8% for B. lonestari, respectively. Although the infection prevalence of B. burgdorferi in I. scapularis was considerably higher than the infection prevalence of B. lonestari in A. americanum, the higher encounter frequencies for A. americanum compared with I. scapularis observed in this and other studies may result in increased risk of acquiring exposure to A. americanum-transmitted pathogens. The potential public health implications of these results are discussed.

Comparative host feeding patterns of the Asian tiger mosquito, Aedes albopictus, in urban and suburban Northeastern USA and implications for disease transmission.

BACKGROUND: Aedes albopictus is an invasive species which continues expanding its geographic range and involvement in mosquito-borne diseases such as chikungunya and dengue. Host selection patterns by invasive mosquitoes are critically important because they increase endemic disease transmission and drive outbreaks of exotic pathogens. Traditionally, Ae. albopictus has been characterized as an opportunistic feeder, primarily feeding on mammalian hosts but occasionally acquiring blood from avian sources as well. However, limited information is available on their feeding patterns in temperate regions of their expanded range. Because of the increasing expansion and abundance of Ae. albopictus and the escalating diagnoses of exotic pathogens in travelers returning from endemic areas, we investigated the host feeding patterns of this species in newly invaded areas to further shed light on its role in disease ecology and assess the public health threat of an exotic arbovirus outbreak. METHODOLOGY/PRINCIPAL FINDINGS: We identified the vertebrate source of 165 blood meals in Ae. albopictus collected between 2008 and 2011 from urban and suburban areas in northeastern USA. We used a network of Biogents Sentinel traps, which enhance Ae. albopictus capture counts, to conduct our collections of blooded mosquitoes. We also analyzed blooded Culex mosquitoes collected alongside Ae. albopictus in order to examine the composition of the community of blood sources. We found no evidence of bias since as expected Culex blood meals were predominantly from birds (n = 149, 93.7%) with only a small proportion feeding on mammals (n = 10, 6.3%). In contrast, Aedes albopictus fed exclusively on mammalian hosts with over 90% of their blood meals derived from humans (n = 96, 58.2%) and domesticated pets (n = 38, 23.0% cats; and n = 24, 14.6% dogs). Aedes albopictus fed from humans significantly more often in suburban than in urban areas (χ(2), p = 0.004) and cat-derived blood meals were greater in urban habitats (χ(2), p = 0.022). Avian-derived blood meals were not detected in any of the Ae. albopictus tested. CONCLUSIONS/SIGNIFICANCE: The high mammalian affinity of Ae. albopictus suggests that this species will be an efficient vector of mammal- and human-driven zoonoses such as La Crosse, dengue, and chikungunya viruses. The lack of blood meals obtained from birds by Ae. albopictus suggest that this species may have limited exposure to endemic avian zoonoses such as St. Louis encephalitis and West Nile virus, which already circulate in the USA. However, growing populations of Ae. albopictus in major metropolitan urban and suburban centers, make a large autochthonous outbreak of an arbovirus such as chikungunya or dengue viruses a clear and present danger. Given the difficulties of Ae. albopictus suppression, we recommend that public health practitioners and policy makers install proactive measures for the imminent mitigation of an exotic pathogen outbreak.

Insecticide resistance status of United States populations of Aedes albopictus and mechanisms involved.

Aedes albopictus (Skuse) is an invasive mosquito that has become an important vector of chikungunya and dengue viruses. Immature Ae. albopictus thrive in backyard household containers that require treatment with larvicides and when adult populations reach pest levels or disease transmission is ongoing, adulticiding is often required. To assess the feasibility of control of USA populations, we tested the susceptibility of Ae. albopictus to chemicals representing the main insecticide classes with different modes of action: organochlorines, organophosphates, carbamates, pyrethroids, insect growth regulators (IGR), naturalytes, and biolarvicides. We characterized a susceptible reference strain of Ae. albopictus, ATM95, and tested the susceptibility of eight USA populations to five adulticides and six larvicides. We found that USA populations are broadly susceptible to currently available larvicides and adulticides. Unexpectedly, however, we found significant resistance to dichlorodiphenyltrichloroethane (DDT) in two Florida populations and in a New Jersey population. We also found resistance to malathion, an organophosphate, in Florida and New Jersey and reduced susceptibility to the IGRs pyriproxyfen and methoprene. All populations tested were fully susceptible to pyrethroids. Biochemical assays revealed a significant up-regulation of GSTs in DDT-resistant populations in both larval and adult stages. Also, ß-esterases were up-regulated in the populations with suspected resistance to malathion. Of note, we identified a previously unknown amino acid polymorphism (Phe → Leu) in domain III of the VGSC, in a location known to be associated with pyrethroid resistance in another container-inhabiting mosquito, Aedes aegypti L. The observed DDT resistance in populations from Florida may indicate multiple introductions of this species into the USA, possibly from tropical populations. In addition, the mechanisms underlying DDT resistance often result in pyrethroid resistance, which would undermine a remaining tool for the control of Ae. albopictus. Continued monitoring of the insecticide resistance status of this species is imperative.

Area-wide ground applications of Bacillus thuringiensis var. israelensis for the control of Aedes albopictus in residential neighborhoods: from optimization to operation.

The increasing range of Aedes albopictus, the Asian tiger mosquito, in the USA and the threat of chikungunya and dengue outbreaks vectored by this species have necessitated novel approaches to control this peridomestic mosquito. Conventional methods such as adulticiding provide temporary relief, but fail to manage this pest on a sustained basis. We explored the use of cold aerosol foggers and misting machines for area-wide applications of Bacillus thuringiensis var. israelensis (VectoBac WDG) as a larvicide targeting Aedes albopictus. During 2010-2013 we performed initially open field trials and then 19 operational area-wide applications in urban and suburban residential areas in northeastern USA to test three truck-mounted sprayers at two application rates. Area-wide applications of WDG in open field conditions at 400 and 800 g/ha killed on average 87% of tested larvae. Once techniques were optimized in residential areas, applications with a Buffalo Turbine Mist Sprayer at a rate of 800 g/ha, the best combination, consistently provided over 90% mortality. Importantly, there was no significant decrease in efficacy with distance from the spray line even in blocks of row homes with trees and bushes in the backyards. Under laboratory conditions Bti deposition in bioassay cups during the operational trials resulted in over 6 weeks of residual control. Our results demonstrate that area-wide truck mounted applications of WDG can effectively suppress Ae. albopictus larvae and should be used in integrated mosquito management approaches to control this nuisance pest and disease vector.

Economic evaluation of an area-wide integrated pest management program to control the Asian tiger mosquito in New Jersey.

Aedes albopictus is the most invasive mosquito in the world, an important disease vector, and a biting nuisance that limits outdoor activities. Area-wide integrated pest management (AW-IPM) is the recommended control strategy. We conducted an economic evaluation of the AW-IPM project in Mercer and Monmouth Counties, New Jersey with a controlled design (AW-IPM vs. control) from 2009 through 2011. The study analyzed financial documents and staff time for AW-IPM and surveyed an average of 415 randomly chosen households in AW-IPM and control areas each fall from 2008 through 2011. Hours lost from yard and porch activities were calculated as differences between actual and potential hours of these activities in an average summer week if there had been no mosquito concerns. Net estimated benefits of AW-IPM were based on cross-over and difference-in-difference analyses. Reductions in hours lost were valued based on respondents' willingness to pay for a hypothetical extra hour free of mosquitoes spent on yard or porch activities and literature on valuation of a quality adjusted life year (QALY). The incremental cost of AW-IPM per adult was $41.18 per year. Number of hours lost due to mosquitoes in AW-IPM areas between the base year (2008) and the intervention years (2009-2011) declined by 3.30 hours per summer week in AW-IPM areas compared to control areas. Survey respondents valued this improvement at $27.37 per adult per summer week. Over the 13-week summer, an average adult resident gained 42.96 hours of yard and porch time, worth $355.82. The net benefit over the summer was $314.63. With an average of 0.0027 QALYs gained per adult per year, AW-IPM was cost effective at $15,300 per QALY gained. The benefit-cost ratio from hours gained was 8.64, indicating that each $1 spent on AW-IPM gave adults additional porch and yard time worth over $8.

Rapid blood meal scoring in anthropophilic Aedes albopictus and application of PCR blocking to avoid pseudogenes.

Blood meal analysis (BMA) is a useful tool for epidemiologists and vector ecologists to assess which vector species are critical to disease transmission. In most current BMA assays vertebrate primers amplify DNA from a blood meal, commonly an abundant mitochondrial (mtDNA) locus, which is then sequenced and compared to known sequences in GenBank to identify its source. This technique, however, is time consuming and costly as each individual sample must be sequenced for species identification and mixed blood meals cloned prior to sequencing. Further, we found that several standard BMA vertebrate primers match sequences of the mtDNA of the Asian tiger mosquito, Aedes albopictus, making their use for blood meal identification in this species impossible. Because of the importance of Ae. albopictus as a vector of dengue and chikungunya viruses to humans, we designed a rapid assay that allows easy identification of human blood meals as well as mixed meals between human and nonhuman mammals. The assay consists of a nested PCR targeting the cytochrome b (cytb) mtDNA locus with a blocking primer in the internal PCR. The blocking primer has a 3' inverted dT modification that when used with the Stoffel Taq fragment prevents amplification of nuclear cytochrome b pseudogenes in humans and allows for the continued use of cytb in BMA studies, as it is one of the most species-rich loci in GenBank. We used our assay to examine 164 blooded specimens of Ae. albopictus from suburban coastal New Jersey and found 62% had obtained blood from humans with 7.6% mixes between human and another mammal species. We also confirmed the efficiency of our assay by comparing it with standard BMA primers on a subset of 62 blooded Ae. albopictus. While this assay was designed for use in Ae. albopictus, it will have broader application in other anthropophilic mosquitoes.