Interspecific Competition between Aedes albopictus and A. sierrensis: potential for Competitive Displacement in the Western United States.

The Asian tiger mosquito, Aedes albopictus, was first detected in North America twenty five years ago. It utilizes water-holding container habitats as immature development sites, and has rapidly spread throughout the eastern United States. Aedes albopictus has occasionally been detected in the western United States, but until recently no established populations of A. albopictus were reported. The western tree-hole mosquito, Aedes sierrensis, is the most common tree-hole mosquito throughout the western United States, and is expected to more frequently encounter A. albopictus. In this study, competition between A. albopictus from the eastern United States and A. sierrensis from the western United States was tested in order to better understand the potential for either competitive displacement of A. sierrensis by A. albopictus or competitive resistance of A. sierrensis to A. albopictus. Varying densities of each species were reared with limited resources in a response surface design. Consistent with a prior study, we found that A. albopictus was clearly a superior larval competitor than A. sierrensis. Aedes sierrensis λ' (finite rate of increase) decreased with increasing A. albopictus density, but in contrast, A. albopictus λ' actually increased with increasing A. sierrensis density; a result that was not reflected by individual fitness parameters. These results indicate that A. sierrensis will not be an effective barrier to A. albopictus invasion into tree-holes in the western United States.

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.

Pesticide-induced release from competition among competing Aedes aegypti and Aedes albopictus (Diptera: Culicidae).

Competitive interactions between mosquitoes Aedes aegypti (L.) and Aedes albopictus (Skuse) may depend on environmental conditions. Pesticides may alleviate density-dependent competition for limited food, and a differential species response to sublethal concentrations may modify interspecific competition. We tested the hypothesis that exposure to malathion alters interspecific resource competition between these two species. In the absence of malathion, Ae. aegypti survivorship and per capita rate of population change were negatively affected by increasing densities of Ae. albopictus. However, the asymmetrical negative effect ofAe. albopictus on Ae. aegypti was eliminated in the presence of malathion. In addition, the presence of malathion resulted in shorter development time compared with the controls. The relative importance of pesticide-mediated coexistence in nature has not been evaluated, so its role in mediating coexistence is unclear; however, these findings underscore the potential of environmental concentrations of malathion, and perhaps other pesticides to facilitate coexistence between species.

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.

Evaluation of a rapid analyte measurement platform for West Nile virus detection based on United States mosquito control programs.

The rapid analyte measurement platform (RAMP) system is an immunoassay test for West Nile virus (WNV) detection. Although reverse transcriptase-polymerase chain reaction (RT-PCR) methodology has been regarded as the gold standard for confirming WNV presence, usage of RAMP testing kits has increased in the past years. We collected RAMP test result data that were subsequently confirmed with RT-PCR methodology from mosquito control agencies to evaluate the efficacy of the RAMP testing. Results indicate that there are a high number of false positives (RAMP positive, RT-PCR negative) with RAMP testing. Correlation between RAMP unit values and RT-PCR cycle threshold values were varied depending on the primer/probe being compared. Comparison of RT-PCR results (on the same samples) between laboratories also indicates variation among the procedures and their potential to influence the RAMP testing efficacies. We discuss the potential issues and solutions that could prevent the high rate of false positives.

New technique to count mosquito adults: using ImageJ software to estimate number of mosquito adults in a trap.

A new technique is described here to count mosquitoes using open-source software. We wanted to develop a protocol that would estimate the total number of mosquitoes from a picture using ImageJ. Adult mosquitoes from CO2-baited traps were spread on a tray and photographed. The total number of mosquitoes in a picture was estimated using various calibrations on ImageJ, and results were compared with manual counting to identify the ideal calibration. The average trap count was 1,541, and the average difference between the manual count and the best calibration was 174.11 +/- 21.59, with 93% correlation. Subsequently, contents of a trap were photographed 5 different times after they were shuffled between each picture to alter the picture pattern of adult mosquitoes. The standard error among variations stayed below 50, indicating limited variation for total count between pictures of the same trap when the pictures were processed through ImageJ. These results indicate the software could be utilized efficiently to estimate total number of mosquitoes from traps.

Efficacy of gravid traps in trapping Culex pipiens.

Members of the Culex pipiens complex are principal vectors for West Nile virus (WNV) in the USA. Previous studies have shown that gravid traps might be more effective than CO2-baited traps in sampling for Cx. pipiens. We compared the efficacy of gravid traps manufactured by 3 different companies: Bioquip, Clarke, and J. W. Hock. All gravid traps have a similar setup to hold the oviposition attractant but differ in the way they collect the mosquitoes. The gravid trap manufactured by J. W. Hock Company trapped significantly more Cx. pipiens than the other traps. Because CO2-baited American Biophysics Corporation (ABC) traps are most often used by mosquito abatement agencies, we compared the efficacy of the Hock gravid trap with a CO2-baited ABC trap. There was no significant difference in the number of Cx. pipiens trapped between the Hock gravid and CO2-baited ABC trap. Because gravid traps predominantly attract previously bloodfed females (thereby aiding in WNV surveillance) and are logistically easier and cheaper to set up, we argue that the Hock gravid trap might be ideal for sampling Cx. pipiens populations.

Behavioral differences of invasive container-dwelling mosquitoes to a native predator.

Aquatic prey show behavioral modifications in the presence of predation-risk cues that alleviate their risk from predation. Aedes albopictus (Skuse), Aedes japonicus (Theobald), and Culex pipiens L. are invasive mosquitoes in North America, and their larvae are prey for the native mosquito predator, Toxorhynchites rutilus (Coquillett). Ae. albopictus and Ae. japonicus are recent invaders, whereas Cx. pipiens has been in the United States for >100 yr. In the presence of predation-risk cues from Tx. rutilus larvae, Cx. pipiens larvae increased the time spent resting at the surface (least risky behavior) more than the other prey species. Ae. japonicus larvae increased resting at the surface of the containers more than Ae. albopictus larvae in the presence of predation-risk cues. Cx. pipiens larvae spent more time motionless at the surface even in the absence of predation-risk cues when compared with the other species, indicating that Cx. pipiens larvae are the least vulnerable prey. As compared with the other prey species, Ae. albopictus larvae exhibited more high-risk behaviors both in the presence and absence of predation-risk cues, indicating that they are the most vulnerable prey. Ae. albopictus is the superior competitor; however, predation by Tx. rutilus larvae may prevent competitive exclusion by Ae. albopictus and promote coexistence among the three prey species.

Can pesticides and larval competition alter susceptibility of Aedes mosquitoes (Diptera: Culicidae) to arbovirus infection?

Density-dependent processes such as larval competition may be important regulatory factors among some mosquito species. The application of pesticides used for control may alter these density-dependent interactions with consequences for the number of survivors and associated sublethal and chronic effects on these individuals. We examined how intraspecific competition among larvae and low concentrations of malathion alter Aedes aegypti L. and Aedes albopictus Skuse adult life history traits and competence for arboviruses using Sindbis virus as a model system. Larvae were reared at densities of 150 and 300 larvae per container and in the absence or presence of 0.04 parts per million of malathion, before surviving females were exposed to an infectious blood meal containing 10(5) plaque-forming units/ml Sindbis virus. For both species, competition and the presence of malathion reduced survival to adulthood. The presence of malathion eliminated the negative effects of competition that resulted in lengthened development time and smaller-sized adults. For Ae. aegypti, but not Ae. albopictus, high competition conditions and the presence of malathion independently and not interactively led to an increase in virus dissemination from the midgut. Our results suggest that larval competition and chemical contaminants may influence disease transmission directly by altering adult mosquito fitness and indirectly by altering vector interactions with arboviruses.

Effect of malathion on larval competition between Aedes albopictus and Aedes atropalpus (Diptera: Culicidae).

Aedes albopictus (Skuse) and Aedes atropalpus (Coquillett) (Diptera: Culicidae) are container-dwelling mosquito species that are well established in the eastern United States. Interspecific larval competition studies have shown Ae. albopictus to be a superior competitor over many species. A laboratory experiment was conducted in artificial containers to evaluate the effects of malathion on larval interactions between Ae. albopictus and Ae. atropalpus. The survivorship of Ae. albopictus increased with increasing Ae. atropalpus densities in control but decreased with increasing Ae. atropalpus densities in the presence of malathion. Alternatively, Ae. atropalpus survivorship did not differ between control and malathion treatments. Developmental times were not affected by interspecific competition in both treatments for either species. These results show that malathion could facilitate coexistence between Ae. albopictus and Ae. atropalpus. This demonstrates how sublethal concentrations of malathion (and perhaps other pesticides with similar modes of action) can enable an inferior competitor to coexist in the same habitat with a superior competitor. This is the first report of synergistic survival of a weaker mosquito competitor in the presence of a pesticide due to condition-specific competition.

Malathion influences competition between Aedes albopictus and Aedes japonicus.

Competitive interactions may facilitate or repel invaders into new communities, and these interactions may depend on other environmental conditions such as the presence of pesticides. Malathion is widely used in controlling agricultural pests and mosquitoes worldwide. Small amounts of malathion, previously considered inconsequential, may in fact increase in lethality when combined with biotic stressors in aquatic systems. We tested whether low concentrations of malathion (0.11 ppm) that are often detected in aquatic systems, affect competition between two invasive mosquito species Aedes albopictus (Skuse) and Aedes japonicus Theobald. There were no survivors of Ae. japonicus larvae in malathion. There was a significant negative effect of Ae. japonicus density on Ae. albopictus survival, but this effect was absent in the presence of malathion. There was also a moderate negative effect of Ae. japonicus density on Ae. albopictus female size, but this effect was absent in the presence of malathion. These findings indicate that pesticide-mediated alterations in competition and species-specific differences in susceptibility to pesticides could play a role in enhancing invasive potential of Ae. albopictus.

Interaction of a pesticide and larval competition on life history traits of Culex pipiens.

Mosquito larval development occurs in aquatic habitats that are directly or indirectly exposed to chemical contaminants. Little is known about how interaction of these chemicals with other biotic and abiotic stressors impact mosquito populations. We used two levels of nutrient (low and high) and four larval densities (10, 20, 30, 40) to examine the effects of low concentrations of insecticide malathion, on Culex pipiens L. mosquitoes experiencing stress from larval competition. Addition of malathion at the high nutrient condition enhanced survival with increasing larval densities, but this effect was not observed at low nutrient condition. Males exposed to malathion were significantly larger than those from control treatments while the effect of malathion on size of females varied with larval density and the level of nutrients. Larval exposure to malathion and low nutrient resulted in significantly larger females with increases in larval densities compared with other treatments. The effect of malathion on male longevity varied with larval density and amount of nutrients. At higher densities, male longevity was consistently higher in low nutrient than in high nutrient conditions and addition of malathion in high nutrient treatment increased male longevity at the highest density. These effects are most likely attributable to release from competition among survivors after mortality from malathion and density-dependent effects. We conclude that biotic conditions of the larval habitat can alter the impact of low concentrations of chemical contaminants on mosquito populations in ways that may influence the pattern of disease transmission and the outcome of vector control efforts.

Suppressing Aedes albopictus, an emerging vector of dengue and chikungunya viruses, by a novel combination of a monomolecular film and an insect-growth regulator.

The Asian tiger mosquito Aedes albopictus (Skuse) is rapidly increasing its global range and importance in transmission of chikungunya and dengue viruses. We tested pellet formulations of a monomolecular film (Agnique) and (S)-methoprene (Altosid) under laboratory and field conditions. In the laboratory, Agnique provided 80% control for 20 days, whereas Altosid, in combination with Agnique, provided 80% control for > 60 days. During field trials, the 1:1 pellet ratio of combined products provided > 95% control for at least 32 days and 50% control for at least 50 days. Altosid remained effective after a 107-day laboratory-induced drought, suggesting that the product serves as a means of control during drought conditions and against spring broods in temperate regions. Agnique and Altosid, when used in tandem for cryptic, difficult-to-treat locations, can provide long-term control of Ae. albopictus larvae and pupae. The possible additive or synergistic effects of the combined products deserve further investigation.

Nature of Predation Risk Cues in Container Systems: Mosquito Responses to Solid Residues From Predation.

In aquatic systems, prey animals associate predation risk with cues that originate either from the predator or from injured conspecifics. Sources and benefits of these cues have received considerable attention in river, lake, and pond ecosystems but are less well understood in small container ecosystems that can hold less than a liter of water. Mosquitoes Aedes triseriatus (Say) and Aedes albopictus (Skuse) encounter predatory Corethrella appendiculata (Grabham) and Toxorhynchites rutilus (Coquillett) in small containers and show antipredatory behavioral responses. We investigated the sources of the predation cues to which these prey larvae respond. We tested whether Ae. albopictus larvae show behavioral responses to cues emanating from the predator or from damage to prey caused by the act of predation. We also tested whether Ae. triseriatus respond to cues present in fluid or solid residues from predator activity. Ae. albopictus showed behavioral modifications only in response to waterborne cues from a feeding predator and not to cues from a starving predator, indicating that Ae. albopictus respond to cues created by the act of predation, which could include substances derived from damaged prey or substances in predator feces. Ae. triseriatus showed behavioral responses to solid residues from predation but not to fluid without those solids, indicating that the cues to which they respond originate in predator feces or uneaten prey body parts. Our results suggest that cues in this system may be primarily chemicals that are detected upon contact with solid residues that are products of the feeding processes of these predators.

An unusual larval collection and survival of Orthopodomyia signifera in the presence of the predator Toxorhynchites rutilus septentrionalis.

From a discarded heavy-equipment tire (ca. 57 liter) at an industrial construction site, we collected 655 (86.0%) Orthopodomyia signifera, 23 (3.0%) Toxorhynchites rutilus septentrionalis, 17 (2.2%) Aedes japonicus japonicus, and 67 (8.8%) Culex pipiens pipiens. Although larvae of Aedes albopictus and Aedes triseriatus were not collected from this container, both species were prevalent as host-seeking adults and readily collected as larvae from other containers at this site. Laboratory trials to test the survival of prey (Ae. albopictus, Cx. p. pipiens, or Or. signifera) in the presence of Tx. rut. septentrionalis showed that survival of prey larvae differed among species. Multiple comparisons revealed that Ae. albopictus had the lowest and Or. signifera the highest survival in the presence of Tx. rut. septentrionalis. Survival of Or. signifera and Cx. p. pipiens was not significantly different from one another, but both were different from Ae. albopictus. Further testing is warranted to test other factors responsible for differences in the interspecific relationship between Or. signifera and other species in tree hole communities.

No evolutionary response to four generations of laboratory selection on antipredator behavior of Aedes albopictus: potential implications for biotic resistance to invasion.

Aedes albopictus (Skuse) is an invasive container-dwelling mosquito and an important disease vector that co-occurs with the native mosquito, Aedes triseriatus (Say), and the predatory midge, Corethrella appendiculata (Grabham). Larval Ae. triseriatus show significantly greater antipredatory responses when compared to larval Ae. albopictus in the presence of predation cues from C. appendiculata. The potential for evolution of antipredatory behavioral responses to C. appendiculata in Ae. albopictus is unknown. We used a controlled laboratory selection experiment to test whether Ae. albopictus could evolve antipredatory behavioral responses to C. appendiculata predation. We subjected replicate Ae. albopictus populations to four generations of predation by C. appendiculata or a predator-free control treatment and compared the behavior and life history of Ae. albopictus in the two treatments in each generation. There were no differences in Ae. albopictus behavioral responses between predation and control lines in any of the four generations. There was also no evidence of differences in life histories between predation and control lines. Ae. albopictus is superior as a competitor compared with Ae. triseriatus, which it has replaced in areas where C. appendiculata are rare. Our results suggest limited potential for Ae. albopictus to evolve stronger antipredatory behavioral responses to C. appendiculata predation and imply that C. appendiculata will continue to act as an impediment to invasion by Ae. albopictus and replacement of Ae. triseriatus and to promote coexistence of these competitors.

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.

Growth and survival of invasive Aedes albopictus larvae on Diospyros virginiana leaves.

Studies on the interactions of exotic species with their invaded environment are imperative in understanding their invasion biology. Larvae of container mosquitoes such as the invasive Aedes albopictus (Skuse) feed on microorganisms that subsist on allochthonous inputs like leaves. Ae. albopictus are vectors for many diseases including West Nile virus and are rapidly expanding their distribution in the United States. We tested the larval performance ofAe. albopictus at different larval densities in maple, oak, American elm, and persimmon. Survival was significantly lower and days to pupation were significantly higher with persimmon leaves compared with all others. In a follow-up experiment, we compared the performance of Ae. albopictus in different amounts of oak and persimmon and different ratios of persimmon + oak. The linear model for the growth rate (defined by larval head width) showed a positive slope as the amount of oak leaves increased in oak treatment, but there was no significant slope for persimmon. In the persimmon + oak combination, as the ratio of persimmon to oak increased, the growth rates of the larvae decreased. Lack of a significant slope for survival rate in combination with the results from the growth rate indicated that persimmon was a poor nutritional resource for Ae. albopictus.

Stage-dependent predation on competitors: consequences for the outcome of a mosquito invasion.

1. Predator-mediated coexistence occurs when predation allows competitors to coexist, due to preferential consumption of a superior competitor relative to an inferior competitor. Differences between the native treehole mosquito (Aedes triseriatus) and the co-occurring Asian tiger mosquito (Aedes albopictus) in anti-predatory larval behaviours account, in part, for the greater vulnerability of this invasive species to native predatory midge (Corethrella appendiculata). We test the hypothesis that stage-dependent differences in the sizes of A. albopictus and A. triseriatus larvae, relative to the size-limited C. appendiculata, contribute to differential consumption and the likelihood of predator-mediated coexistence of these competitors. 2. In all instars, larvae of A. triseriatus were larger than A. albopictus of the same stage. Third and fourth instar C. appendiculata selectively consumed late-stage A. albopictus in preference to same-stage A. triseriatus. Small, early-stage prey larvae did not differ in vulnerability to predation, but large, late-stage larvae differed significantly in vulnerability to predation, probably owing to size-limited predation by fourth instar C. appendiculata. This effect was less pronounced for third instar C. appendiculata. 3. Prey size, in conjunction with anti-predatory behavioural responses, alters the probability of predator-mediated coexistence. A stage-structured predation model showed that equally vulnerable early stages reduce the range of environmental conditions (productivities) in which predator-mediated coexistence is possible, increasing the likelihood of both competitive exclusion of the resident species or failure of the invasive to establish. These results underscore the importance of stage-dependent interspecific differences in predator-prey interactions for determining how predators may affect community composition.

Do natural container habitats impede invader dominance? Predator-mediated coexistence of invasive and native container-dwelling mosquitoes.

Predator-mediated coexistence of competitors occurs when a species that is superior in competition is also more vulnerable to a shared predator compared to a poorer competitor. The invasive mosquito Aedes albopictus is usually competitively superior to Ochlerotatus triseriatus. Among second instar larvae, A. albopictus show a lesser degree of behavioral modification in response to water-borne cues from predation by the larval midge Corethrella appendiculata than do O. triseriatus, rendering A. albopictus more vulnerable to predation by C. appendiculata than O. triseriatus. The hypothesis that C. appendiculata predation favors coexistence of these competitors predicts that C. appendiculata abundances will be negatively and positively correlated with A. albopictus and O. triseriatus abundances, respectively, and that coexistence will occur where C. appendiculata are common. Actual abundances of O. triseriatus, A. albopictus, and C. appendiculata in three habitats fit this prediction. In natural container habitats like tree holes, C. appendiculata were abundant and competitors co-existed at similar densities. In cemeteries and tires, which occur primarily in non-forested, human-dominated habitats, A. albopictus dominated, with abundances twice those found in tree holes, but C. appendiculata and O. triseriatus were rare or absent. We also tested for whether antipredatory behavioral responses of A. albopictus differed among habitats or populations, or were correlated with local C. appendiculata abundances. We could detect no differences in A. albopictus antipredatory behavioral responses to water-borne cues from predation. Tree hole habitats appear to promote co-existence of O. triseriatus and A. albopictus through interactions with predatory C. appendiculata, and this predator effect appears to limit invasion success of A. albopictus in tree holes. There are many studies on predator-mediated coexistence in natural habitats but to our knowledge this is the first study to suggest differential predator-mediated coexistence between natural and man-made habitats.