Suppression of Midgut Microbiota Impact Pyrethroid Susceptibility in Aedes aegypti.

Aedes aegypti is a mosquito that transmits viral diseases such as dengue, chikungunya, Zika, and yellow fever. The insect's microbiota is recognized for regulating several biological processes, including digestion, metabolism, egg production, development, and immune response. However, the role of the bacteria involved in insecticide susceptibility has not been established. Therefore, the objective of this study was to characterize the resident microbiota in a field population of A. aegypti to evaluate its role associated with susceptibility to the insecticides permethrin and deltamethrin. Mosquitoes were fed 10% sucrose mixed with antibiotics and then exposed to insecticides using a diagnostic dose. DNA was extracted, and sequencing of bacterial 16S rRNA was carried out on Illumina® MiSeq™. Proteobacteria (92.4%) and Bacteroidetes (7.6%) were the phyla, which are most abundant in mosquitoes fed with sucrose 10%. After exposure to permethrin, the most abundant bacterial species were Pantoea agglomerans (38.4%) and Pseudomonas azotoformans-fluorescens-synxantha (14.2%). Elizabethkingia meningoseptica (38.4%) and Ps. azotoformans-fluorescens-synxantha (26.1%) were the most abundant after exposure to deltamethrin. Our results showed a decrease in mosquitoes' survival when exposed to permethrin, while no difference in survival when exposed to deltamethrin when the microbiota was modified. We found that the change in microbiota modifies the response of mosquitoes to permethrin. These results are essential for a better understanding of mosquito physiology in response to insecticides.

Determination of Knockdown Resistance (kdr) Allele Frequencies (T929I mutation) in Head Louse Populations from Mexico, Canada, and Peru.

The head louse Pediculus humanus capitis (De Geer) is a hematophagous ectoparasite that inhabits the human scalp. The infestations are asymptomatic; however, skin irritation from scratching occasionally may cause secondary bacterial infections. The present study determined the presence and frequency of the knockdown resistance (kdr) mutation T929I in 245 head lice collected from Mexico, Peru, and Canada. Head lice were collected manually using a comb in the private head lice control clinic. Allele mutation at T9291 was present in 100% of the total sampled populations (245 lice) examined. In addition, 4.89% of the lice were homozygous susceptible, whereas 6.93% heterozygous and 88.16% homozygous were resistant, respectively. This represents the second report in Mexico and Quebec and fist in Lima.

Field Effectiveness of Drones to Identify Potential Aedes aegypti Breeding Sites in Household Environments from Tapachula, a Dengue-Endemic City in Southern Mexico.

Aedes aegypti control programs require more sensitive tools in order to survey domestic and peridomestic larval habitats for dengue and other arbovirus prevention areas. As a consequence of the COVID-19 pandemic, field technicians have faced a new occupational hazard during their work activities in dengue surveillance and control. Safer strategies to monitor larval populations, in addition to minimum householder contact, are undoubtedly urgently needed. Drones can be part of the solution in urban and rural areas that are dengue-endemic. Throughout this study, the proportion of larvae breeding sites found in the roofs and backyards of houses were assessed using drone images. Concurrently, the traditional ground field technician's surveillance was utilized to sample the same house groups. The results were analyzed in order to compare the effectiveness of both field surveillance approaches. Aerial images of 216 houses from El Vergel village in Tapachula, Chiapas, Mexico, at a height of 30 m, were obtained using a drone. Each household was sampled indoors and outdoors by vector control personnel targeting all the containers that potentially served as Aedes aegypti breeding sites. The main results were that the drone could find 1 container per 2.8 found by ground surveillance; however, containers that were inaccessible by technicians in roofs and backyards, such as plastic buckets and tubs, disposable plastic containers and flowerpots were more often detected by drones than traditional ground surveillance. This new technological approach would undoubtedly improve the surveillance of Aedes aegypti in household environments, and better vector control activities would therefore be achieved in dengue-endemic countries.

First Record of Aedes podographicus in Nuevo León State, Mexico.

In Mexico, Aedes podographicus is one of the most common species within the subgenus Protomacleaya of Aedes. This species has been collected in 12 states close to the coastal regions; however, few records confirm the presence of Ae. podographicus inland. During a mosquito-survey using ovitraps in the state of Nuevo León, Mexico, Ae. podographicus was collected in association with Ae. albopictus and Culex quinquefasciatus. This is the first record of the presence of Ae. podographicus in Nuevo León. With the addition of Ae. podographicus to the mosquito fauna of Nuevo León, there are currently 66 species in the state, 19 within the genus Aedes and 6 within the subgenus Protomacleaya, the genus Aedes being the group with the major number of species in Nuevo León. Specimens collected during this study were deposited in the collection of insects and mites of medical importance of the Laboratory of Medical Entomology of the Autonomous University of Nuevo León, Mexico.

Identifying urban hotspots of dengue, chikungunya, and Zika transmission in Mexico to support risk stratification efforts: a spatial analysis.

BACKGROUND: Effective Aedes aegypti control is limited, in part, by the difficulty in achieving sufficient intervention coverage. To maximise the effect of vector control, areas with persistently high numbers of Aedes-borne disease cases could be identified and prioritised for preventive interventions. We aimed to identify persistent Aedes-borne disease hotspots in cities across southern Mexico. METHODS: In this spatial analysis, geocoded cases of dengue, chikungunya, and Zika from nine endemic Mexican cities were aggregated at the census-tract level. We included cities that were located in southern Mexico (the arbovirus endemic region of Mexico), with a high burden of dengue cases (ie, more than 5000 cases reported during a 10-year period), and listed as high priority for the Mexican dengue control and prevention programme. The Getis-Ord Gi*(d) statistic was applied to yearly slices of the dataset to identify spatial hotspots of each disease in each city. We used Kendall's W coefficient to quantify the agreement in the distribution of each virus. FINDINGS: 128 507 dengue, 4752 chikungunya and 25 755 Zika clinical cases were reported between Jan 1, 2008, and Dec 31, 2016. All cities showed evidence of transmission heterogeneity, with a mean of 17·6% (SD 4·7) of their total area identified as persistent disease hotspots. Hotspots accounted for 25·6% (SD 9·7; range 12·8-43·0) of the population and 32·1% (10·5; 19·6-50·5) of all Aedes-borne disease cases reported. We found an overlap between hotspots of 61·7% for dengue and Zika and 53·3% for dengue and chikungunya. Dengue hotspots in 2008-16 were significantly associated with dengue hotspots detected during 2017-20 in five of the nine cities. Heads of vector control confirmed hotspot areas as problem zones for arbovirus transmission. INTERPRETATION: This study provides evidence of the overlap of Aedes-borne diseases within geographical hotspots and a methodological framework for the stratification of arbovirus transmission risk within urban areas, which can guide the implementation of surveillance and vector control. FUNDING: USAID, the US Centers for Disease Control and Prevention, the Canadian Institutes of Health Research, International Development Research Centre, Fondo Mixto CONACyT (Mexico)-Gobierno del Estado de Yucatan, and the US National Institutes of Health. TRANSLATION: For the Spanish translation of the abstract see Supplementary Materials section.

miRNAs of Aedes aegypti (Linnaeus 1762) conserved in six orders of the class Insecta.

Aedes aegypti L. is the most important vector of arboviruses such as dengue, Zika, chikungunya, Mayaro, and yellow fever, which impact millions of people's health per year. MicroRNA profile has been described in some mosquito species as being important for biological processes such as digestion of blood, oviposition, sexual differentiation, insecticide resistance, and pathogens dissemination. We identified the miRNAs of Ae. aegypti females, males and eggs of a reference insecticide susceptible strain New Orleans and compared them with those other insects to determine miRNA fingerprint by new-generation sequencing. The sequences were analyzed using data mining tools and categorization, followed by differential expression analysis and conservation with other insects. A total of 55 conserved miRNAs were identified, of which 34 were of holometabolous insects and 21 shared with hemimetabolous insects. Of these miRNAs, 32 had differential expression within the stages analyzed. Three predominant functions of miRNA were related to embryonic development regulation, metamorphosis, and basal functions. The findings of this research describe new information on Ae. aegypti physiology which could be useful for the development of new control strategies, particularly in mosquito development and metamorphosis processes.

Efficacy of Long-lasting Insecticidal Nets With Declining Physical and Chemical Integrity on Aedes aegypti (Diptera: Culicidae).

Fitting long-lasting insecticidal nets (LLIN) as screens on doors/windows has a significant impact on indoor-adult Aedes aegypti (L.), with entomological reductions measured in a previous study being significant for up to 2 yr post-installation, even in the presence of pyrethroid-resistant Aedes populations. To better understand the mode of LLIN protection, bioassays were performed to evaluate the effects of field deployment (0, 6, and 12 mo) and damage type (none, central, lateral, and multiple) on LLIN efficacy. Contact bioassays confirmed that LLIN residual activity (median knockdown time, in minutes, or MKDT) decreased significantly over time: 6.95 (95% confidence interval [CI]: 5.32-8.58) to 9.24 (95% CI: 8.69-9.79) MKDT at 0- and 12-mo age, respectively, using a pyrethroid-susceptible Aedes strain. Tunnel tests (exposing human forearm for 40 min as attractant) showed that deployment time affected negatively Aedes passage inhibition from 54.9% (95% CI: 43.5-66.2) at 0 mo to 35.7% (95% CI: 16.3-55.1) at 12 mo and blood-feeding inhibition from 65.2% (95% CI: 54.2-76.2) to 48.9% (95% CI: 26.4-71.3), respectively; both the passage/blood-feeding inhibition increased by a factor of 1.8-2.9 on LLINs with multiple and central damages compared with nets with lateral damage. Mosquito mortality was 74.6% (95% CI: 65.3-83.9) at 0 mo, 72.3% (95% CI: 64.1-80.5) at 6 mo, and 59% (95% CI: 46.7-71.3) at 12 mo. Despite the LLIN physical integrity could be compromised over time, we demonstrate that the remaining chemical effect after field conditions would still contribute to killing/repelling mosquitoes.

New Record of Aedes albopictus In A Suburban Area Of Merida, Yucatan, Mexico.

A survey was carried out in 51 households within a suburban area of Merida, Yucatan, Mexico, for 5 consecutive days. Adult collections were performed using Prokopack aspirators (indoors) and human-landing mosquito catches (HLC) outdoors, and larval sites (artificial containers) were revised for larvae collection. A total of 259 Aedes albopictus were collected, 246 from artificial larval sites, 8 by indoor aspiration, and 5 by HLC. This is the first record of Ae. albopictus in Merida, Yucatan, Mexico.

Variation in competence for ZIKV transmission by Aedes aegypti and Aedes albopictus in Mexico.

BACKGROUND: ZIKV is a new addition to the arboviruses circulating in the New World, with more than 1 million cases since its introduction in 2015. A growing number of studies have reported vector competence (VC) of Aedes mosquitoes from several areas of the world for ZIKV transmission. Some studies have used New World mosquitoes from disparate regions and concluded that these have a variable but relatively low competence for the Asian lineage of ZIKV. METHODOLOGY/PRINCIPAL FINDINGS: Ten Aedes aegypti (L) and three Ae. albopictus (Skuse) collections made in 2016 from throughout Mexico were analyzed for ZIKV (PRVABC59-Asian lineage) VC. Mexican Ae. aegypti had high rates of midgut infection (MIR), dissemination (DIR) and salivary gland infection (SGIR) but low to moderate transmission rates (TR). It is unclear whether this low TR was due to heritable salivary gland escape barriers or to underestimating the amount of virus in saliva due to the loss of virus during filtering and random losses on surfaces when working with small volumes. VC varied among collections, geographic regions and whether the collection was made north or south of the Neovolcanic axis (NVA). The four rates were consistently lower in northeastern Mexico, highest in collections along the Pacific coast and intermediate in the Yucatan. All rates were lowest north of the NVA. It was difficult to assess VC in Ae. albopictus because rates varied depending upon the number of generations in the laboratory. CONCLUSIONS/SIGNIFICANCE: Mexican Ae. aegypti and Ae. albopictus are competent vectors of ZIKV. There is however large variance in vector competence among geographic sites and regions. At 14 days post infection, TR varied from 8-51% in Ae. aegypti and from 2-26% in Ae. albopictus.

Report of Chikungunya Virus in Wild Populations of Aedes aegypti in Guerrero State, Mexico.

We detected vertical transmission of chikungunya virus (CHIKV) in wild populations of Aedes aegypti from San Marcos, Guerrero, Mexico, with real-time reverse transcriptase-polymerase chain reaction. A total of 20 pools (1-11 specimens/pool) of larvae, male, and female mosquitoes were tested. We report the detection of CHIKV in 2 of 11 larval pools, 4 of 5 male pools, and 1 of 4 female pools, from field-collected mosquitoes.

Profiles of Amino Acids and Acylcarnitines Related with Insecticide Exposure in Culex quinquefasciatus (Say).

Culex quinquefasciatus Say is a vector of many pathogens of humans, and both domestic and wild animals. Personal protection, reduction of larval habitats, and chemical control are the best ways to reduce mosquito bites and, therefore, the transmission of mosquito-borne pathogens. Currently, to reduce the risk of transmission, the pyrethroids, and other insecticide groups have been extensively used to control both larvae and adult mosquitoes. In this context, amino acids and acylcarnitines have never been associated with insecticide exposure and or insecticide resistance. It has been suggested that changes in acylcarnitines and amino acids profiles could be a powerful diagnostic tool for metabolic alterations. Monitoring these changes could help to better understand the mechanisms involved in insecticide resistance, complementing the strategies for managing this phenomenon in the integrated resistance management. The purpose of the study was to determine the amino acids and acylcarnitines profiles in larvae of Cx. quinquefasciatus after the exposure to different insecticides. Bioassays were performed on Cx. quinquefasciatus larvae exposed to the diagnostic doses (DD) of the insecticides chlorpyrifos (0.001 µg/mL), temephos (0.002 µg/mL) and permethrin (0.01 µg/mL). In each sample, we analyzed the profile of 12 amino acids and 31 acylcarnitines by LC-MS/MS. A t-test was used to determine statistically significant differences between groups and corrections of q-values. Results indicates three changes, the amino acids arginine (ARG), free carnitine (C0) and acetyl-carnitine (C2) that could be involved in energy production and insecticide detoxification. We confirmed that concentrations of amino acids and acylcarnitines in Cx. quinquefasciatus vary with respect to different insecticides. The information generated contributes to understand the possible mechanisms and metabolic changes occurring during insecticide exposure.

[Effectiveness of commercial repellents against Aedes aegypti (L.) in Yucatan, México]. / Efectividad de repelentes comerciales disponibles contra el mosquito Aedes aegypti (L.) en Yucatán, México.

OBJECTIVE: We assessed the efficacy of commercial repellents available in Yucatan against Aedes aegypti, vector of dengue, Chikungunya and Zika. MATERIALS AND METHODS: Protection time was determined based on WHO/CTD/ WHOPES/IC y la NOM-032-SSA2-20I4. RESULTS: Products with DEET (N, N-diethyl-3-methylbenzamide) at 25% met the recommended protection. Efficacy was directly proportional to the concentration of DEET; botanicals repellents resulted no protective. CONCLUSIONS: Repellents with DEET provided more protection against Ae. aegypti and botanical repellents, including impregnated wristbands, provided no protection.

Efectividad de repelentes comerciales disponibles contra el mosquito Aedes aegypti (L ) en Yucatán, México / Effectiveness of commercial repellents against Aedes aegypti (L ) in Yucatan, México

Resumen: Objetivo: Determinar la eficacia de repelentes comerciales disponibles en Yucatán contra el mosquito Aedes aegypti, vector del dengue, Chikungunya y Zika. Material y métodos: Se determinó el tiempo de protección con base en el protocolo WHO/CTD/WHOPES/IC y la NOM-032-SSA2-2014. Resultados: Sólo el repelente con DEET (N, N-dietil-3-metilbenzamida) al 25% cumplió con la protección recomendada. La eficacia fue directamente proporcional a la concentración del DEET; aquéllos con componentes botánicos fueron poco o nada protectores. Conclusiones: Los resultados muestran que los repelentes con DEET proveen protección contra Ae. Aegypti; los repelentes botánicos, incluyendo las pulseras impregnadas, ofrecen nula protección.

Abstract: Objective: We assessed the efficacy of commercial repellents available in Yucatan against Aedes aegypti, vector of dengue, Chikungunya and Zika. Materials and methods: Protection time was determined based on WHO/CTD/ WHOPES/IC y la NOM-032-SSA2-20I4. Results: Products with DEET (N, N-diethyl-3-methylbenzamide) at 25% met the recommended protection. Efficacy was directly proportional to the concentration of DEET; botanicals repellents resulted no protective. Conclusions: Repellents with DEET provided more protection against Ae. aegypti and botanical repellents, including impregnated wristbands, provided no protection.

Mecanismos asociados a la resistencia al derribo "kdr" a la deltametrina en Aedes aegypti del occidente de Venezuela / Mechanisms associated with knockdown resistance to deltamethrin in Aedes aegypti from western Venezuela

Se determinaron los mecanismos bioquímicos y moleculares involucrados con la resistencia al derribo "kdr" a la deltametrina en poblaciones de Aedes aegypti de los estados Trujillo, Lara y Táchira. Las poblaciones fueron expuestas a CK50 previamente determinadas mediante bioensayos con botellas impregnadas siguiendo la metodología de Brogdon (1989) por 1h. Posteriormente los insectos fueron colocados en envases post-recuperación libres de insecticidas y separados en 4 fenotipos: los no derribados luego de 1h, los recuperados a las 4h, los supervivientes y los muertos a las 24 horas post-exposición. Todos los ejemplares fueron seccionados; con cabeza y tórax se determinaron los niveles de esterasas α y β, oxidasas de función múltiple, glutation S transferasas y acetilcolinesterasa insensible y con el abdomen se extrajo ADN y se realizaron PCR para amplificar los alelos específicos Val1016 e Ile1016. Las enzimas desintoxicantes se incrementaron en la mayoría de las poblaciones entre las 4 y 24h posteriores a la exposición a la deltametrina sin encontrarse diferencia significativa con los niveles expresados en la cepa susceptible New Orleans (NO), excepto en la población de Ureña donde se encontró aumento significativo en las β-esterasas siendo superiores en el fenotipo superviviente con respecto al fenotipo muertos a las 24h. El genotipo silvestre V1016/V1016 prevaleció sobre el heterocigoto y homocigoto mutante en los cuatro fenotipos, en la mayoría de las poblaciones estudiadas, con excepción de la población Ureña donde el homocigoto mutante I1016/I1016 fue el genotipo predominante en los no derribados, lo cual se vio reflejado en la frecuencia alélica. Se asocia la mutación V1016I con la resistencia al derribo mostrada en las poblaciones evaluadas, destacando la importancia de la temprana detección de esta y otras mutaciones en el canal del sodio asociadas con resistencia a piretroides, lo cual debe ser considerado antes de incorporar el uso de deltametrina en el programa de control de Ae. aegypti en estas poblaciones.

The biochemical and molecular mechanisms associated with resistance to deltamethrin were determined in female Aedes aegypti taken from different mosquito populations captured in Trujillo, Lara and Tachira states. Individuals from each population were subjected to 1 h of exposure to deltamethrin using the CK50 previously determined by the bottle bioassay. The mosquitoes were then placed in containers free from insecticide and separated into 4 phenotypes: mosquitoes that were not knocked down after 1 h of exposure, those that recovered 4 h after exposure, those that were still alive 24 h after exposure and those that were dead at 24 h. Each of the mosquitoes in these groups was then dissected to separate the head-thorax, and abdomen. Biochemical tests were performed on the head-thorax to determine the presence of resistance-related enzymes including: α-and β-esterases, glutathione S-transferase and insensitive acetylcholinesterase. The abdomen was used for molecular tests to amplify the specific allele Val 1016 and Ile 1016. The quantities of detoxifying enzymes increased between 4 and 24 h after exposure to deltamethrin in mosquitoes from most of the populations tested although no significant differences between these and the susceptible New Orleans strain (NO) were found, except for mosquitoes from the Ureña population which showed a significant increase in β-esterase with higher values in the "survivors" phenotype compared to the "dead" phenotype at 24 h. The wild genotype V1016/V1016 prevailed over the heterozygous and homozygous mutants in the four phenotypes in the majority of the populations studied, with exception of the Ureña population where the resistant homozygote I1016/I1016 was the predominant genotype. The V1016I mutation was associated with the knockdown resistance observed in the evaluated populations emphasizing the importance of the early detection of this and other mutations in the sodium channel which have been linked with resistance to pyrethroids. These aspects should be considered before applying deltamethrin to control these Ae. Aegypti populations.