Evaluación de la sensibilidad a organofosforados en poblaciones de Aedes aegypti (L. ) (Diptera: Culicidae) del departamento de La Guajira, Colombia / Evaluation of susceptibility to organophosphates in populations of Aedes aegypti (L. ) (Diptera: Culicidae) in La Guajira, Colombia

Introducción. El dengue es un problema de salud pública para el departamento de La Guajira. El control se ha enfocado en el vector con el uso de insecticidas, entre ellos los organofosforados. Objetivo. Evaluar el estado de la sensibilidad a insecticidas organofosforados de quince poblaciones de Aedes aegypti (L.) en el departamento de La Guajira, Colombia. Materiales y métodos. Se realizaron bioensayos para temefos, malatión y pirimifos- metil en larvas de tercer estadio y mosquitos adultos de Ae. aegypti en los municipios de Albania, Barrancas, Dibulla, Distracción, El Molino, Fonseca, Hatonuevo, La Jagua del Pilar, Maicao, Manaure, Riohacha, San Juan del Cesar, Uribia, Urumita y Villanueva, siguiendo la metodología de la Organización Mundial de la Salud (OMS) y la técnica de botellas usando la guía de los de los Centers for Disease Control and Prevention, respectivamente. Se determinó la sensibilidad por medio de la relación de resistencia a CL50 y CL95 (RRCL50, RRCL95) para temefos y a dosis y tiempo diagnóstico para temefos, malatión y pirimifos-metil en las poblaciones de campo evaluadas, usando como control la cepa sensible Rockefeller. Resultados. Las 15 poblaciones del departamento de La Guajira son sensibles a: temefos (relación de la resistencia a CL50<5,0; relación de resistencia a CL95<5,0; 98 a 100 % de mortalidad); pirimifos-metil (99 a 100 % de mortalidad) y malatión (100 % de mortalidad). Conclusión. Con base en los resultados obtenidos, es factible el uso de temefos, malatión y pirimifos-metil para el control de Ae. aegypti en las poblaciones evaluadas.

Introduction. Dengue is a public health problem in La Guajira region. Control has focused on the vector using insecticides, including organophosphates. Objective. To evaluate the state of susceptibility to organophosphates insecticides in fifteen Aedes aegypti (L.) populations in La Guajira, Colombia. Materials and methods. We collected samples of third-instar larvae and adult mosquitoes of Ae. aegypti in the municipalities of Albania, Barrancas, Dibulla, Distracción, El Molino, Fonseca, Hatonuevo, La Jagua del Pilar, Maicao, Manaure, Riohacha, San Juan del Cesar, Uribia, Urumita, Villanueva. Bioassays for temefos, malathion, and pirimiphos-methyl were carried out following the methodology of the World Health Organization, and the bottle technique using the guidance of the Centers for Disease Control and Prevention. Susceptibility to temefos was determined through the resistance ratio between lethal concentration 50 and lethal concentration 95; for the compounds temefos, malathion and pirimiphos-methyl, susceptibility was calculated using diagnostic dose and diagnostic time in the populations evaluated. Rockefeller susceptible strain was used as a control. Results. All evaluated populations of Ae. aegypti from La Guajira were found to be susceptible to temefos (ratio resistance to CL50<5.0; ratio resistance to CL95<5.0; 98 - 100 % mortality); pirimiphosmethyl (99 - 100 % mortality), and malathion (100 % mortality). Conclusion. Based on the results, the use of temefos, malathion, and pirimiphosmethyl is feasible for the control of Ae. aegypti in the evaluated populations.

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.

Molecular Characterization of Associated Pathogens in Febrile Patients during Inter-Epidemic Periods of Urban Arboviral Diseases in Tapachula Southern Mexico.

Emerging and re-emerging vector-borne infections are a global public health threat. In endemic regions, fever is the main reason for medical attention, and the etiological agent of such fever is not usually identified. In this study, non-specific febrile pathogens were molecularly characterized in serum samples from 253 patients suspected of arbovirus infection. The samples were collected in the southern border region of Mexico from April to June 2015, and February to March 2016. ZIKV, CHIKV, DENV, leptospirosis, and rickettsiosis were detected by qPCR and nested PCR to identify flavivirus and alphavirus genera. The results indicated that 71.93% of the samples were positive for CHIKV, 0.79% for ZIKV, and 0.39% for DENV, with the number positive for CHIKV increasing to 76.67% and those positive for ZIKV increasing to 15.41% under the nested PCR technique. Leptospira Kmetyi was identified for the first time in Mexico, with a prevalence of 3.16%. This is the first report of ZIKV in Mexico, as well the first detection of the virus in early 2015. In conclusion, the etiological agent of fever was determined in 94% of the analyzed samples.

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.

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.

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.

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.

[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 de resistencia a la permetrina en dos poblaciones de Aedes aegypti del occidente de Venezuela / Mechanisms of Permethrin resistance in two populations of Aedes aegypti in westernVenezuela

Se determinó la resistencia a la permetrina en Aedes aegypti de los estados Trujillo y Zulia mediante bioensayos de botella. Los insectos derribados durante la hora de exposición fueron registrados y usados para calcular la Concentración Knock-down cincuenta (CK50) y los muertos a las 24 horas para la Concentración Letal cincuenta(CL50). La resistencia al derribo y post-recuperación fueron determinadas calculando del Factor de Resistencia FRCK50 y FRCL50, comparando los valores de CK50 y CL50de las poblaciones de Ae. aegypti de campo con los de la cepa susceptible New Orleans obtenidos mediante análisis de regresión log-probit. Mecanismos metabólicos y no metabólicos asociados a la resistencia, fueron evaluados midiendo los niveles de las enzimas alfa-esterasas, beta-esterasas, oxidasas de función múltiple y glutatión-S-transferasas mediante la técnica de microplacas y determinando la frecuencia alélica I1016 por PCR alelo específico. Ambas poblaciones mostraron baja resistencia al derribo (FRCK50 < 5) y moderada resistencia post-recuperación (FRCL50 entre 5 y 10). Sobre-expresión de alfa-esterasas fue observada en la población Loma Linda la cual se correlacionó significativamente con la CL50. En la población Pampanito la frecuencia del alelo I1016 fue de 0,1 y en Loma Linda de 0,17, observándose homocigotos mutantes solo en esta última población. Se evidencia la presencia de mecanismos metabólicos y no metabólicos asociados a la resistencia al derribo y post-recuperación a la permetrina en las poblaciones bajo estudio, lo cual debe ser considerado antes de la aplicación de piretroides para el control de Ae. aegypti en la zona de estudio.

Permethrin resistance was determined in Aedes aegypti populations from Trujillo and Zulia states using the bottle bioassay method. Insect knock-down rates during 1h of exposure were recorded and used to calculate the 50% knock-down concentration (KC50) and the mortality after 24 h (LC50). Knock-down and post-recovery resistance were determined by calculating the resistance factors, FRKC50 and FRLC50 . This was done by comparing the KC50 and LC50 values (obtained by regression analysis log-probit) of the field populations with a susceptible New Orleans strain. Metabolic and non-metabolic mechanisms associated with resistance were assessed by measuring the levels of the following enzymes: alpha-esterases, beta-esterases, mixed function oxidases (MFOs) and glutathione-S-transferases (GSTs) using the microplate technique. We also determined the alellic frequency of I1016 by allele specific PCR. Both populations showed a low knock-down resistance (FRKC50 < 5) and moderate post-recovery resistance (FRLC50 between 5 and 10). Overexpression of alpha-esterases was observed in the Loma Linda population and was significantly correlated with the LC50 . The frequency of the I1016 allele was 0.1 for the Pampanito population and 0.17 for the Loma Linda population, and in the latter we also observed homozygous mutants. The existence of metabolic and non-metabolic mechanisms associated with knockdown resistance and post-recovery to permethrin in the populations studied was demonstrated. This should be taken into account before introducing these insecticides to control populations of Ae. aegypti in the region.

Local evolution of pyrethroid resistance offsets gene flow among Aedes aegypti collections in Yucatan State, Mexico.

The mosquito Aedes aegypti is the major vector of the four serotypes of dengue virus (DENV1-4). Previous studies have shown that Ae. aegypti in Mexico have a high effective migration rate and that gene flow occurs among populations that are up to 150 km apart. Since 2000, pyrethroids have been widely used for suppression of Ae. aegypti in cities in Mexico. In Yucatan State in particular, pyrethroids have been applied in and around dengue case households creating an opportunity for local selection and evolution of resistance. Herein, we test for evidence of local adaptation by comparing patterns of variation among 27 Ae. aegypti collections at 13 single nucleotide polymorphisms (SNPs): two in the voltage-gated sodium channel gene para known to confer knockdown resistance, three in detoxification genes previously associated with pyrethroid resistance, and eight in putatively neutral loci. The SNPs in para varied greatly in frequency among collections, whereas SNPs at the remaining 11 loci showed little variation supporting previous evidence for extensive local gene flow. Among Ae. aegypti in Yucatan State, Mexico, local adaptation to pyrethroids appears to offset the homogenizing effects of gene flow.

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.

Larvicidal and cytotoxic activities of extracts from 11 native plants from northeastern Mexico.

Of all mosquito-borne viral diseases, dengue is spreading most rapidly worldwide. Conventional chemical insecticides (e.g., organophosphates and carbamates) effectively kill mosquitoes at their larval stage, but are toxic to humans. Natural product-based insecticides may be highly specific. Herein, we report the insecticidal activities of 11 native Mexican plants against Aedes aegypti (L). Ether extracts of Ambrosia confertiflora De Candolle, Thymus vulgaris (L.), and Zanthoxylum fagara (L.), and both ether and methanol extracts of Ruta chalepensis L. were significantly larvicidal toward the dengue mosquito after 24 h of exposure. Of them, only the ether extract of A. confertiflora was toxic to Vero cells. In conclusion, the ether extracts of Thymus vulgaris, Z. fagara, and both ether and methanol extracts of Ruta chalepensis L., could be considered as potential bioinsecticides.

Gonotrophic cycle and survivorship of Culex quinquefasciatus (Diptera: Culicidae) using sticky ovitraps in Monterrey, northeastern Mexico.

Mark-release-recapture experiments were conducted to determine the length of the gonotrophic cycle and rate of survivorship of Culex quinquefasciatus Say in Monterrey, northeastern Mexico. A total of 2,352 field-caught Cx. quinquefasciatus females were marked and released at 8-12 h postemergence in 2 field trials. Sticky ovitraps were used to recapture marked gravid females. One hundred and ten (4.6%) marked females were recaptured during a 12-day sampling period. Recapture rates for the 2 individual trials were 6.4% and 3.5%. The length of the gonotrophic cycle, calculated as the average time between the initial blood meal and the time of recapture of gravid females, was 2-3 days. The first blood-fed mosquitoes were recaptured on the 2nd day postrelease. Gravid egg-laying females were most commonly recaptured at 2-3 days postfeeding. Daily survival estimates for the 2 release dates were of 0.871 and 0.883, respectively.

Host-feeding preference of Culex quinquefasciatus in Monterrey, northeastern Mexico.

Studies were conducted to determine the host selection patterns of Culex quinquefasciatus in the municipalities of Guadalupe and Escobedo near Monterrey, northeastern Mexico. Mosquitoes were captured inside and outside houses. Chickens and humans were the most common blood sources for all Cx. quinquefasciatus females, accounting for nearby 70% of blood meals. Human blood was detected by enzyme-linked immunosorbent assay in 36.4% and 28.4% of engorged females resting inside houses in Guadalupe and Escobedo, respectively. The proportions of indoor resting females fed on chicken blood were 38.7% and 56.7%, respectively. The weighted and unweighted human blood index (HBI) values were calculated, by using indoor and outdoor data, from the proportions of humanfed mosquitoes. Weighted means (HBI) estimates for Guadalupe and Escobedo were 23.0% and 15.4%, respectively. The forage ratios (FRs) for humans were <1.0 (with or without chicken populations); consequently, it seems that these mosquitoes feed on humans with less frequency in comparison with chickens, horses, and pigs. The FRs for chickens were the highest of all available hosts (1.7 and 3.2), and they were the most abundant hosts in Escobedo, and the second most abundant in Guadalupe, indicating a selective bias of Cx. quinquefasciatus for chickens (i.e. ornithophagic).