The L1014F Knockdown Resistance Mutation Is Not a Strong Correlate of Phenotypic Resistance to Pyrethroids in Florida Populations of Culex quinquefasciatus.

Culex quinquefasciatus is an important target for vector control because of its ability to transmit pathogens that cause disease. Most populations are resistant to pyrethroids and often to organophosphates, the two most common classes of active ingredients used by public health agencies. A knockdown resistance (kdr) mutation, resulting in an amino acid change from a leucine to phenylalanine in the voltage gated sodium channel, is one mechanism contributing to the pyrethroid resistant phenotype. Enzymatic resistance has also been shown to play a very important role. Recent studies have shown strong resistance in populations even when kdr is relatively low, which indicates that factors other than kdr may be larger contributors to resistance. In this study, we examined, on a statewide scale (over 70 populations), the strength of the correlation between resistance in the CDC bottle bioassay and the kdr genotypes and allele frequencies. Spearman correlation analysis showed only moderate (-0.51) or weak (-0.29) correlation between the kdr genotype and permethrin or deltamethrin resistance, respectively. The frequency of the kdr allele was an even weaker correlate than genotype. These results indicate that assessing kdr in populations of Culex quinquefasciatus is not a good surrogate for phenotypic resistance testing.

Trends in mosquito species distribution modeling: insights for vector surveillance and disease control.

Species distribution modeling (SDM) has become an increasingly common approach to explore questions about ecology, geography, outbreak risk, and global change as they relate to infectious disease vectors. Here, we conducted a systematic review of the scientific literature, screening 563 abstracts and identifying 204 studies that used SDMs to produce distribution estimates for mosquito species. While the number of studies employing SDM methods has increased markedly over the past decade, the overwhelming majority used a single method (maximum entropy modeling; MaxEnt) and focused on human infectious disease vectors or their close relatives. The majority of regional models were developed for areas in Africa and Asia, while more localized modeling efforts were most common for North America and Europe. Findings from this study highlight gaps in taxonomic, geographic, and methodological foci of current SDM literature for mosquitoes that can guide future efforts to study the geography of mosquito-borne disease risk.

Spatiotemporal Environmental Drivers of Eastern Equine Encephalitis Virus in Central Florida: Towards a Predictive Model for a Lethal Disease.

Eastern equine encephalitis virus (EEEV) is a mosquito-borne virus that affects humans and horses, with a high case fatality rate in both species. The virus can be transmitted by several mosquito species and maintained in multiple reservoir hosts, making EEEV dynamics difficult to anticipate. In this study, we identified spatial and temporal factors that precede EEEV detection using sentinel chicken surveillance data from Orange County, Florida, from 2003 to 2017. We first examined the land cover and mosquito species composition associated with sentinel chicken sites. We then fit distributed lag nonlinear models of EEEV detection at the county scale, using monthly temperature, precipitation, and Southern Oscillation Index values, and at the sentinel flock-scale, using remotely sensed temperature and wetness indicators. We found positive associations between the percent wooded wetlands and the count of EEEV detections. We found Culiseta melanura (Diptera: Culicidae) were more abundant at positive sites in winter and summer, but Coquillettidia perturbans (Walker) were more abundant at positive sites in spring. In the county-wide model, precipitation, temperature, and Southern Oscillation Index values at lags of two, nine, and twelve months were significant, respectively, while temperature and wetness were significant at lags of eight and six months in the flock-specific models.

Examining Wing Length-Abundance Relationships and Pyrethroid Resistance Mutations among Aedes albopictus in a Rapidly Growing Urban Area with Implications for Mosquito Surveillance and Control.

Aedes albopictus is a cosmopolitan mosquito species capable of transmitting arboviruses such as dengue, chikungunya, and Zika. To control this and similar species, public and private entities often rely on pyrethroid insecticides. In this study, we screened Ae. albopictus collected from June to August 2017 in Mecklenburg County, a rapidly growing urban area of North Carolina, for mutations conferring pyrethroid resistance and examined spatiotemporal patterns of specimen size as measured by wing length, hypothesizing that size variation could be closely linked to local abundance, making this easily measured trait a useful surveillance proxy. The genetic screening results indicated that pyrethroid resistance alleles are not present in this population, meaning that this population is likely to be susceptible to this commonly used insecticide class. We detected no significant associations between size and abundance-related factors, indicating that wing-size is not a useful proxy for abundance, and thus not useful to surveillance in this capacity. However, mosquitoes collected in June were significantly larger than July or August, which may result from meteorological conditions, suggesting that short-term weather cues may modulate morphological traits, which could then affect local fecundity and virus transmission dynamics, as previously reported.

Spatial variation in the frequency of knockdown resistance genotypes in Florida Aedes aegypti populations.

BACKGROUND: The development of insecticide resistance in disease-vectoring mosquito species can lead to vector control failure and disease resurgence. However, insecticide applications remain an essential public health intervention. In Florida, insecticide resistance in Aedes aegypti, an anthropophilic mosquito species capable of transmitting dengue, chikungunya, and Zika virus, is a major concern. Understanding the location, scale, and driving factors of insecticide resistance can enhance the ability of vector control organizations to target populations effectively. METHODS: We used previously collected data on frequencies of mutations that confer resistance to commonly used pyrethroid insecticides in Ae. aegypti specimens from 62 sites distributed across 18 counties in Florida. To determine the scale of clustering for the most resistant variant, we used a Ripley's K function. We also used a spatial scanning statistic technique to identify locations of clusters where higher than expected frequencies of susceptible or resistant mosquitoes occurred. We then tested for associations between landscape, demographic, and insecticide-use factors using a beta regression modelling approach and evaluated the effect of spatial lag and spatial error terms on overall explanatory power of these models. RESULTS: The scale at which maximum clustering of the most resistant variant occurs is approximately 20 kilometers. We identified statistically significant clusters of genotypes associated with resistance in several coastal cities, although some of these clusters were near significant clusters of susceptible mosquitoes, indicating selection pressures vary at the local scale. Vegetation density, distance from roads, and pyrethroid-use by vector control districts were consistently significant predictors of knockdown resistance genotype frequency in the top-performing beta regression models, although pyrethroid use surprisingly had a negatively associated with resistance. The incorporation of spatial lags resulted in improvements to the fit and explanatory power of the models, indicating an underlying diffusion process likely explains some of the spatial patterns observed. CONCLUSIONS: The genetic mutations that confer resistance to pyrethroids in Ae. aegypti mosquitoes in Florida exhibit spatial autocorrelation and patterns that can be partially explained by landscape and insecticide-use factors. Further work at local scales should be able to identify the mechanisms by which these variables influence selection for alleles associated with resistance.

Loxodonta Localizer: A Software Tool for Inferring the Provenance of African Elephants and Their Ivory Using Mitochondrial DNA.

Illegal hunting is a major threat to the elephants of Africa, with more elephants killed by poachers than die from natural causes. DNA from tusks has been used to infer the source populations for confiscated ivory, relying on nuclear genetic markers. However, mitochondrial DNA (mtDNA) sequences can also provide information on the geographic origins of elephants due to female elephant philopatry. Here, we introduce the Loxodonta Localizer (LL; www.loxodontalocalizer.org), an interactive software tool that uses a database of mtDNA sequences compiled from previously published studies to provide information on the potential provenance of confiscated ivory. A 316 bp control region sequence, which can be readily generated from DNA extracted from ivory, is used as a query. The software generates a listing of haplotypes reported among 1917 African elephants in 24 range countries, sorted in order of similarity to the query sequence. The African locations from which haplotype sequences have been previously reported are shown on a map. We demonstrate examples of haplotypes reported from only a single locality or country, examine the utility of the program in identifying elephants from countries with varying degrees of sampling, and analyze batches of confiscated ivory. The LL allows for the source of confiscated ivory to be assessed within days, using widely available molecular methods that do not depend on a particular platform or laboratory. The program enables identification of potential regions or localities from which elephants are being poached, with capacity for rapid identification of populations newly or consistently targeted by poachers.

Seasonal and geographic variation in insecticide resistance in Aedes aegypti in southern Ecuador.

Insecticide resistance (IR) can undermine efforts to control vectors of public health importance. Aedes aegypti is the main vector of resurging diseases in the Americas such as yellow fever and dengue, and recently emerging chikungunya and Zika fever, which have caused unprecedented epidemics in the region. Vector control remains the primary intervention to prevent outbreaks of Aedes-transmitted diseases. In many high-risk regions, like southern Ecuador, we have limited information on IR. In this study, Ae. aegypti IR was measured across four cities in southern Ecuador using phenotypic assays and genetic screening for alleles associated with pyrethroid IR. Bottle bioassays showed significant inter-seasonal variation in resistance to deltamethrin, a pyrethroid commonly used by the Ministry of Health, and alpha-cypermethrin, as well as between-city differences in deltamethrin resistance. There was also a significant difference in phenotypic response to the organophosphate, Malathion, between two cities during the second sampling season. Frequencies of the resistant V1016I genotype ranged from 0.13 to 0.68. Frequencies of the resistant F1534C genotype ranged from 0.63 to 1.0, with sampled populations in Machala and Huaquillas at fixation for the resistant genotype in all sampled seasons. In Machala and Portovelo, there were statistically significant inter-seasonal variation in genotype frequencies for V1016I. Resistance levels were highest in Machala, a city with hyperendemic dengue transmission and historically intense insecticide use. Despite evidence that resistance alleles conferred phenotypic resistance to pyrethroids, there was not a precise correspondence between these indicators. For the F1534C gene, 17.6% of homozygous mutant mosquitoes and 70.8% of heterozygotes were susceptible, while for the V1016I gene, 45.6% homozygous mutants and 55.6% of heterozygotes were susceptible. This study shows spatiotemporal variability in IR in Ae. aegypti populations in southern coastal Ecuador, and provides an initial examination of IR in this region, helping to guide vector control efforts for Ae. aegypti.