Effects of a Red Marker Dye on Aedes and Culex Larvae: Are There Implications for Operational Mosquito Control?

Marker dyes are often mixed with liquid insecticide formulations prior to field applications to accurately determine the characteristics and penetration of droplets into targeted habitats. We have been using FD&C Red 40 Granular DM food dye at the rate of 20 g/liter in liquid solutions of Bacillus thuringiensis israelensis (Bti) for area-wide larvicide applications against the Asian tiger mosquito, Aedes albopictus. The Bti and dye mix ratio has been recommended by pesticide manufacturers for testing under operational conditions, but no data exist on the effects of the dye itself on mosquito larvae. We tested the effects of the FD&C Red 40 food dye in laboratory bioassays against different strains of Ae. albopictus (New Jersey and Maryland) and Culex pipiens pipiens (Utah) at rates of 0.039 to 80.0 g/liter. We also conducted field application trials to measure dye concentrations up to 100 m downwind when mixed and applied according to manufacturer instructions. In laboratory bioassays, we found that mean survival in cups with dye were significantly different from the controls beginning at 10.0 g/liter for New Jersey Ae. albopictus and at 20.0 g/liter for Maryland Ae. albopictus and Utah Cx. p. pipiens. In field application trials, we recorded a maximum volume density of 1,152.8 nl/cm(2) and calculated the maximum concentration of dye at 9.09 × 10(-3) g/liter. Our results showed that although we detected greater effects of dye on Ae. albopictus in New Jersey experiments than Ae. albopictus in Maryland and Cx. p. pipiens from Utah, concentrations of the dye during operational applications were at least 1,100 times below concentrations that exhibited toxic effects for either species in the laboratory, suggesting that the dye will not interfere with accuracy of field bioassays. Our results conclusively demonstrate that the addition of the FD&C Red 40 marker dye does not alter the efficacy of the pesticide formulation by skewing results, but rather provides a valuable addition to accurately determine pesticide penetration and spectrum by discriminating between intended pesticide and other potential pollutants.

Evaluation of ultra-low volume treatments on Culex quinquefasciatus (Diptera: Culicidae) abundance and relative age by parity dissection.

St. Tammany Parish Mosquito Abatement District relies on ultra-low volume (ULV) insecticides to manage adult Culex quinquefasciatus (Say 1823), the primary vector of West Nile virus in Louisiana, but abundance rarely declines post-treatment. Three field ULV applications each of adulticides containing deltamethrin, naled, or resmethrin were conducted from May through October 2022 to measure efficacy against free-flying Cx. quinquefasciatus. Female mosquitoes trapped before and after treatment were identified, counted, and age-graded by parity dissection. No insecticide applications reduced Cx. quinquefasciatus abundance, but trials with deltamethrin and naled significantly lowered average mosquito age in the area post-treatment. No trials with resmethrin reduced mosquito age, which corroborates laboratory susceptibility data indicating local resistance toward the pyrethroid. These trials establish parity dissections and relative age gradation as a means of measuring treatment success against Cx. quinquefasciatus.

Battle of the Bites: The Effect of Sewage Effluent Exposure on Mosquitofish Biocontrol of Mosquitoes in Residential Louisiana.

Mosquitofish, Gambusia affinis, are eponymous larval mosquito predators. Their ability to colonize and survive in habitats that are uninhabitable by other potential predators allows them to naturally manage larval mosquito populations in most ground pools they are present in. However, effluent from residential onsite wastewater treatment systems (OWTSs) appears to limit the presence of fish predators. This is especially problematic in Louisiana, where regulations allow the discharge of OWTS effluent into open drainage conveyances. To determine the effect of effluent on the capacity of mosquitofish for biocontrol in contaminated areas, we assessed the body condition metrics of populations from two effluent-exposed sites and two sites not exposed to effluent, determined the lethal effect of effluent-contaminated drainage water on fish, and measured the prey consumption rates in the presence of effluent. Female fish collected from effluent-impacted sites had a reduced somatic body condition and most females examined displayed masculinized anal fins resembling the male gonopodium structure. This trait was not seen in fish collected from the control sites and has not yet been documented in association with OWTSs or in the state of Louisiana. Fish from the control sites survived at effluent-contaminated water levels < 70%, and the prey clearance rates increased with dilution. Onsite wastewater treatment system effluent has significant effects on both the short- and long-term persistence of mosquitofish, their body composition, reproductive health, and larval mosquito consumption. These effects likely release mosquito larvae from suppression and may increase the threat of mosquito-transmitted pathogens in effluent-contaminated locations.

An Efficient Alternative to the CDC Gravid Trap for Southern House Mosquito (Diptera: Culicidae) Surveillance.

West Nile virus (WNV) is the most prevalent arbovirus found throughout the United States. Surveillance of surface breeding Culex vectors involved in WNV transmission is primarily conducted using CDC Gravid traps. However, anecdotal claims from mosquito abatement districts in Louisiana assert that other trap types may be more suited to WNV surveillance. To test the validity of these assertions, we conducted a series of trapping trials and WNV surveillance over 3 yr to compare the efficacy of multiple trap types. First, we compared the CDC Gravid trap, CO2-baited New Standard Miniature Blacklight traps, and CO2-baited CDC light traps with either an incandescent light, a red light, or no light. We found that the CDC Gravid trap and CO2-baited no-light CDC Light trap collected the most mosquitoes. Second, we conducted additional, long-term trapping and WNV surveillance to compare these two trap types. We found that CO2-baited no-light CDC traps collected more of the local WNV vector, Culex quinquefasciatus (Say, Diptera, Culicidae), and detected WNV with greater sensitivity. Finally, we conducted trapping to compare the physiological states of Cx. quinquefasciatus and diversity of collected mosquitoes. CO2-baited no-light CDC light traps collected more unfed Cx. quinquefasciatus while Gravid traps collected more blooded Cx. quinquefasciatus; both traps collected the same number of gravid Cx. quinquefasciatus. Additionally, we found that CO2-baited no-light CDC light traps collected a larger diversity of mosquito species than Gravid traps.

Effects of truck-mounted, ultra low volume mosquito adulticides on honey bees (Apis mellifera) in a suburban field setting.

Few studies have examined the impact of mosquito adulticides on honey bees under conditions that reflect actual field exposure. Whereas several studies have evaluated the toxicity of mosquito control products on honey bees, most have been laboratory based and have focused solely on acute mortality as a measure of impact. The goal of this study was to determine effects of routine applications of truck-based ultra-low volume (ULV) mosquito adulticides (i.e., Scourge, Duet, and Deltagard) on honey bees in a suburban setting. The mosquito adulticides used in this study were pyrethroids with active ingredients resmethrin (Scourge), prallethrin and sumithrin (Duet), and deltamethrin (Deltagard), in which resmethrin, prallethrin, and sumithrin were synergized with piperonyl butoxide. We measured and compared mortality and detoxification enzyme activities (esterase and glutathione S-transferase) from sentinel beehives within and outside of mosquito control areas. Concurrently, colony health (i.e., number of adult bees, brood quantity and brood quality) was compared throughout the study period. No significant differences were observed in honey bee mortality, colony health or detoxification enzyme activities between treated (five sprayed areas each received one to three insecticide treatment) and control sites (four unsprayed areas that did not receive insecticide treatment) over the seven week study period. However, our laboratory study showed that exposure to resmethrin, the active ingredient in Scourge, caused significant inhibition of esterase activity compared with the control group. Our findings suggest that proper application of truck based insecticides for mosquito control results in little or no exposure and therefore minimal effects on domestic honey bees.

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.