Impact of the V410L kdr mutation and co-occurring genotypes at kdr sites 1016 and 1534 in the VGSC on the probability of survival of the mosquito Aedes aegypti (L.) to Permanone in Harris County, TX, USA.

Harris County, TX, is the third most populous county in the USA and upon detection of arboviruses Harris County Public Health applies insecticides (e.g., pyrethroid-based Permanone 31-66) against adults of Culex quinquefasciatus to prevent disease transmission. Populations of Aedes aegypti, while not yet a target of public health control, are likely affected by pyrethroid exposure. As this species is a vector of emerging arboviruses, its resistance status to Permanone and the kdr mutations in the voltage-gated sodium channel (VGSC) associated with pyrethroid resistance were investigated. We examined females of known genotype at the V1016I and F1534C sites (N = 716) for their genotype at the 410 amino acid position in the VGSC, and for the influence of their kdr genotype on survival to Permanone at three different distances from the insecticide source in field tests. Most females (81.8%) had at least one resistant L allele at the 410 position, being the first report of the V410L mutation in Ae. aegypti for Texas. When only genotypes at the 410 position were analyzed, the LL genotype exhibited higher survivorship than VL or VV. Out of 27 possible tri-locus kdr genotypes only 23 were found. Analyses of the probability of survival of tri-locus genotypes and for the V410L genotype using a multivariate logistic regression model including area, distance, and genotype found significant interactions between distance and genotype. When only the most common tri-locus genotypes were analyzed (LL/II/CC, 48.2%; VL/II/CC, 19.1%; and VV/II/CC, 10.1%) genotype had no effect on survival, but significant interactions of distance and genotype were found. This indicated that the V410L kdr allele increased survival probability at certain distances. Genotypes did not differ in survivorship at 7.62-m, but LL/II/CC had higher survivorship than VL/II/CC at 15.24- and 22.86-m. The model also identified differences in survivorship among the operational areas investigated.

Kdr genotyping (V1016I, F1534C) of the Nav channel of Aedes aegypti (L.) mosquito populations in Harris County (Houston), Texas, USA, after Permanone 31-66 field tests and its influence on probability of survival.

Aedes aegypti (L.) is an important mosquito vector of emerging arboviruses such as Zika, dengue, yellow fever, and chikungunya. To quell potential disease outbreaks, its populations are controlled by applying pyrethroid insecticides, which selection pressure may lead to the development of insecticide resistance. Target site insensitivity to pyrethroids caused by non-synonymous knockdown resistance (kdr) mutations in the voltage-gated sodium (NaV) channel is a predominant mechanism of resistance in mosquitoes. To evaluate the potential impact of pyrethroid resistance on vector control, Ae. aegypti eggs were collected from eight mosquito control operational areas in Harris County, Texas, and emerged females were treated in field tests at four different distances from the pyrethroid Permanone 31-66 source. The females were genotyped by melting curve analyses to detect two kdr mutations (V1016I and F1534C) in the NaV channel. Harris County females had higher survivorship rates at each distance than the pyrethroid-susceptible Orlando strain females. Survivorship increased with distance from the pyrethroid source, with 39% of field-collected mosquitoes surviving at 7.62 m and 82.3% at 22.86 m from the treatment source. Both the V1016I and F1534C pyrethroid resistant genotypes were widely distributed and at high frequency, with 77% of the females being double homozygous resistant (II/CC), this being the first report of kdr mutations in Ae. aegypti in Harris County. Analysis of the probability of survival for each mutation site independently indicated that the CC genotype had similar probability of survival as the FC heterozygous, while the II genotype had higher survival than both the VI and VV, that did not differ. The double homozygous resistant genotype (II/CC) had the highest probability of survival. A linear model estimated probability of survival for areas and genotypes. The high frequency and widespread distribution of double-homozygote pyrethroid-resistant Ae. aegypti may jeopardize disease vector control efforts in Harris County.

A proposed framework for the development and qualitative evaluation of West Nile virus models and their application to local public health decision-making.

West Nile virus (WNV) is a globally distributed mosquito-borne virus of great public health concern. The number of WNV human cases and mosquito infection patterns vary in space and time. Many statistical models have been developed to understand and predict WNV geographic and temporal dynamics. However, these modeling efforts have been disjointed with little model comparison and inconsistent validation. In this paper, we describe a framework to unify and standardize WNV modeling efforts nationwide. WNV risk, detection, or warning models for this review were solicited from active research groups working in different regions of the United States. A total of 13 models were selected and described. The spatial and temporal scales of each model were compared to guide the timing and the locations for mosquito and virus surveillance, to support mosquito vector control decisions, and to assist in conducting public health outreach campaigns at multiple scales of decision-making. Our overarching goal is to bridge the existing gap between model development, which is usually conducted as an academic exercise, and practical model applications, which occur at state, tribal, local, or territorial public health and mosquito control agency levels. The proposed model assessment and comparison framework helps clarify the value of individual models for decision-making and identifies the appropriate temporal and spatial scope of each model. This qualitative evaluation clearly identifies gaps in linking models to applied decisions and sets the stage for a quantitative comparison of models. Specifically, whereas many coarse-grained models (county resolution or greater) have been developed, the greatest need is for fine-grained, short-term planning models (m-km, days-weeks) that remain scarce. We further recommend quantifying the value of information for each decision to identify decisions that would benefit most from model input.

Detection of the Nav channel kdr-like mutation and modeling of factors affecting survivorship of Culex quinquefasciatus mosquitoes from six areas of Harris County (Houston), Texas, after permethrin field-cage tests.

Culex quinquefasciatus is one of the most important mosquito vectors of arboviruses. Currently, the fastest approach to control disease transmission is the application of synthetic adulticide insecticides. However, in highly populated urban centers the development of insecticide resistance in mosquito populations could impair insecticide efficacy and therefore, disease control. To assess the effect of resistance on vector control, females of Cx. quinquefasciatus collected from six mosquito control operational areas in Harris County, Texas, were treated in field cage tests at three different distances with the pyrethroid Permanone® 31-66 applied at the operational rate. Females were analyzed by sequencing and/or diagnostic PCR using de novo designed primers for detecting the kdr-like mutation in the voltage-gated sodium channel (L982F; TTA to TTT) (house fly kdr canonical mutation L1014F). Females from the Cx. quinquefasciatus susceptible Sebring strain and those from the six operational areas placed at 30.4 m from the treatment source were killed in the tests, while 14% of field-collected mosquitoes survived at 60.8 m, and 35% at 91.2 m from the source. The diagnostic PCR had a with 97.5% accuracy to detect the kdr-like mutation. Pyrethroid resistant mosquitoes carrying the L982F mutation were broadly distributed in Harris County at high frequency. Among mosquitoes analyzed (n = 1,028), the kdr-kdr genotype was prevalent (81.2%), the kdr-s genotype was 18%, and s-s mosquitoes were less than 1% (n = 8). A logistic regression model estimated an equal probability of survival for the genotypes kdr-kdr and kdr-s in all areas analyzed. Altogether, our results point to a high-risk situation for the pyrethroid-based arboviral disease control in Harris County.

Harris County Public Health Mosquito and Vector Control Division Emergency Response to Hurricane Harvey: Vector-Borne Disease Surveillance and Control.

Hurricane Harvey made a landfall on the Texas Gulf Coast on August 25, 2017, stalling over Harris County as a tropical storm for 4 days (August 26-29), dumping approximately 127 cm of rain. This tremendous amount of rainfall overwhelmed the county's natural and man-made drainage systems, resulting in unprecedented widespread flooding. Immediately following, Harris County Public Health Mosquito and Vector Control Division conducted a countywide emergency vector control response by integrating surveillance, control, and education strategies. This included landing rate counts, mosquito and avian surveillance, arbovirus testing, ground-based ultra-low volume (ULV) and aerial pesticide spraying, and community outreach. The immediate response lasted for 4 wk through September, resulting in 774 landing rates, 49,342 ha treated by ground-based ULV, 242,811 ha treated by aerial ULV, 83,241 mosquitoes collected, 1,807 mosquito pools tested, and 20 education/outreach sessions. Recovery activities of 3 additional education/outreach events continued through October while surveillance and control activities returned to routine status.

Microbiome Interaction Networks and Community Structure From Laboratory-Reared and Field-Collected Aedes aegypti, Aedes albopictus, and Culex quinquefasciatus Mosquito Vectors.

Microbial interactions are an underappreciated force in shaping insect microbiome communities. Although pairwise patterns of symbiont interactions have been identified, we have a poor understanding regarding the scale and the nature of co-occurrence and co-exclusion interactions within the microbiome. To characterize these patterns in mosquitoes, we sequenced the bacterial microbiome of Aedes aegypti, Ae. albopictus, and Culex quinquefasciatus caught in the field or reared in the laboratory and used these data to generate interaction networks. For collections, we used traps that attracted host-seeking or ovipositing female mosquitoes to determine how physiological state affects the microbiome under field conditions. Interestingly, we saw few differences in species richness or microbiome community structure in mosquitoes caught in either trap. Co-occurrence and co-exclusion analysis identified 116 pairwise interactions substantially increasing the list of bacterial interactions observed in mosquitoes. Networks generated from the microbiome of Ae. aegypti often included highly interconnected hub bacteria. There were several instances where co-occurring bacteria co-excluded a third taxa, suggesting the existence of tripartite relationships. Several associations were observed in multiple species or in field and laboratory-reared mosquitoes indicating these associations are robust and not influenced by environmental or host factors. To demonstrate that microbial interactions can influence colonization of the host, we administered symbionts to Ae. aegypti larvae that either possessed or lacked their resident microbiota. We found that the presence of resident microbiota can inhibit colonization of particular bacterial taxa. Our results highlight that microbial interactions in mosquitoes are complex and influence microbiome composition.

Field cage insecticide resistance tests against Culex quinquefasciatus Say (Diptera: Culicidae) in Harris County, Texas, U.S.A.

The ground adulticiding program in densely populated Harris County has been employed on the basis of virus-positive mosquitoes almost exclusively using the 'rotation of chemicals best practices' stratagem. To evaluate its effectiveness, 15 comparative field cage testing events were conducted from 2011-2015 using seven wild population samples from repeatedly collected locations of Culex quinquefasciatus Say and a laboratory susceptible Sebring strain colony reared to adults. A 3 × 3 plot design was employed for exposure to ultra-low volume applications of malathion and synergized permethrin. No significant differences were found in mortality rate among testing dates, year, row placement, or relative humidity. Mortality was significantly different between adulticides (p<0.001) with mean mortality rates for malathion 96.42% (±7.95%) and permethrin 92.38% (±14.04). There was a significant temperature difference for permethrin (p<0.001) but none for malathion (p=0.644). Mosquito population mortality was statistically different by study operational area (p<0.011) and chemical (p<0.001). Susceptible colony adults used as positive controls downwind strongly aided determination of efficacy and resistance to each adulticide, providing evidence of individual application coverage, though comparative analysis was done with overall mortalities by normal methodology.

Efficacy of Deltagard®, Fyfanon®, and Evoluer™ 31-66 Against Culex quinquefasciatus In Harris County, Texas.

The comparative efficacy of ultra-low volume applications of deltamethrin, permethrin, and malathion against specific adult populations of Culex quinquefasciatus , a primary regional vector of Saint Louis encephalitis and West Nile virus, using field cage tests is described. At labeled application rates, the estimated mass median diameters for Fyfanon®, Evoluer™, and DeltaGard® were 18.32, 19.24, and 17.44 µm, respectively. No significant differences (P ≥ 0.01) in mean percent control were observed across all 3 wild populations and a susceptible Sebring strain tested with Fyfanon, DeltaGard, and Evoluer, providing 98.25%, 98.20%, and 97.24% mean percent control. Mean percent control for Sebring and 3 wild populations was 99.38%, 97.79%, 97.26%, and 97.15%, respectively, which were not significantly different from one another (P ≥ 0.01). Across all tests, highly significant differences (P ≤ 0.01) in mean percent control were obtained at 30.48, 60.96, and 91.44 m distances downwind, with mean percent control at 91.44 m being significantly less than that observed at 30.48 m, but not at 60.96 m. Individually, tests for DeltaGard, Fyfanon, and Evoluer resulted in no significant differences (P ≥ 0.01) in mean percent control between all mosquito populations tested, ranging from 96.47 to 100. However, Evoluer obtained significantly less (P ≤ 0.01) mean percent control at 91.44 m compared to DeltaGard and Fyfanon. Data suggest that the wild Cx. quinquefasciatus specimens collected were as susceptible as the laboratory Sebring strain. The use of Sebring as positive controls downwind strongly supported determination of the efficacy of all adulticides and susceptibility of wild populations tested.

Efficacy of Two Larvasonic™ Units Against Culex Larvae and Effects on Common Aquatic Nontarget Organisms in Harris County, Texas.

The Larvasonic™ Field Arm Mobile Wetlands Unit and SD-Mini were tested for efficacy against Culex larvae, and effects on aquatic nontarget organisms (NTO). The Field Arm provided 84.61% to 100% control of caged Culex larvae out to 0.91-m distance in shallow ditches and 60.45% control of Culex larvae at 0.61-m without any effects to caged NTO. Slow ditch treatment achieved 77.35% control compared to fast treatment (20.42%), whereas 77.65% control was obtained along edges of a neglected swimming pool, compared to near the middle (23.97%). In bucket tests, the SD-Mini provided >97% control of Culex and 85.35% reduction of immature giant water bugs, which decreased slightly (83.45%) over the monitoring period, which was not significantly different from cannibalistic damselflies (62.80%), with reduction of both being significantly higher than other NTO tested. There was a small (0.37%) reduction of dragonflies (naiads), due to cannibalism. Both Larvasonic units could effectively augment conventional larvicide operations in smaller areas without causing resistance within mosquito populations or harming NTO when used properly.

Thermal fog efficacy tests against mosquitoes in storm drains in Harris County, Texas 2008-09.

In 2008 and 2009, our current thermal fogging methods and materials were evaluated in underground systems against feral and laboratory strains of adult Culex quinquefasciatus. Culex quinquefasciatus adults collected from storm drain systems the night before treatment were tested concurrently with adults from a susceptible laboratory strain (Sebring) in 10 separate tests. During 2008, there were no significant differences in the low mean percent control obtained between any of the feral populations (29.39%) and susceptible (Sebring) populations (56.04%) tested, whereas in 2009, application of mineral oil alone yielded extremely low, but significantly different mean percent control at 0.99% for ferals and 0.01% for Sebrings. During 2009 mineral oil evaluations, 45,677 droplets were collected in storm drains at distances of 99.1, 50.6, 57.9, 67.7, and 109.7 m from the application site, with 99% of the droplets below 3 microm in diam; additionally, we found no significant differences between mean percent control of Sebring and feral mosquito populations using the higher (3.2x) 1:10 application rate of Pyrocide. However, mean percent control between the feral and susceptible strain (Sebring) during 2009 was lower than in 2008 at 16.55% for ferals and 24.43% in Sebrings. Results indicated that control methodologies and/or chemicals used were ineffective at controlling Cx. quinquefasciatus in storm drains using the chosen experimental design. Based upon this information, thermal fog operations were discontinued due to lack of effectiveness.