Adriana Flores Suarez

Quantitative Trait Loci Mapping of Genome Regions Controlling Permethrin Resistance in the Mosquito Aedes aegypti

The mosquito Aedes aegypti is the principal vector of dengue and yellow fever flaviviruses. Permethrin is an insecticide used to suppress Ae. aegypti adult populations but metabolic and target site resistance to pyrethroids has evolved in many locations worldwide. Quantitative trait loci (QTL) controlling permethrin survival in Ae. aegypti were mapped in an F3 advanced intercross line. Parents came from a collection of mosquitoes from Isla Mujeres, México, that had been selected for permethrin resistance for two generations and a reference permethrin-susceptible strain originally from New Orleans. Following a 1-hr permethrin exposure, 439 F3 adult mosquitoes were phenotyped as knockdown resistant, knocked down/recovered, or dead. For QTL mapping, single nucleotide polymorphisms (SNPs) were identified at 22 loci with potential antixenobiotic activity including genes encoding cytochrome P450s (CYP), esterases (EST), or glutathione transferases (GST) and at 12 previously mapped loci. Seven antixenobiotic genes mapped to chromosome I, six to chromosome II, and nine to chromosome III. Two QTL of major effect were detected on chromosome III. One corresponds with a SNP previously associated with permethrin resistance in the para sodium channel gene and the second with the CCEunk7o esterase marker. Additional QTL but of relatively minor effect were also found. These included two sex-linked QTL on chromosome I affecting knockdown and recovery and a QTL affecting survival and recovery. On chromosome II, one QTL affecting survival and a second affecting recovery were detected. The patterns confirm that mutations in the para gene cause target-site insensitivity and are the major source of permethrin resistance but that other genes dispersed throughout the genome contribute to recovery and survival of mosquitoes following permethrin exposure.

Transcription of detoxification genes after permethrin selection in the mosquito Aedes aegypti.

Changes in gene expression before, during and after five generations of permethrin laboratory selection were monitored in six strains of Aedes aegypti: five F2–F3 collections from the Yucatán Peninsula of Mexico and one F2 from Iquitos, Peru. Three biological replicate lines were generated for each strain. The response to selection was measured as changes in the lethal and knockdown permethrin concentrations (LC50, KC50) and in the frequency of the Ile1,016 substitution in the voltage‐gated sodium channel (para) gene. Changes in expression of 290 metabolic detoxification genes were measured using the ‘Aedes Detox’ microarray. Selection simultaneously increased the LC50, KC50 and Ile1,016 frequency. There was an inverse relationship between Ile1,016 frequency and the numbers of differentially transcribed genes. The Iquitos strain lacked the Ile1,016 allele and 51 genes were differentially transcribed after selection as compared with 10–18 genes in the Mexican strains. Very few of the same genes were differentially transcribed among field strains but 10 cytochrome P450 genes were upregulated in more than one strain. Laboratory adaptation to permethrin in Ae. aegypti is genetically complex and largely conditioned by geographic origin and pre‐existing target site insensitivity in the para gene. The lack of uniformity in the genes that responded to artificial selection as well as differences in the direction of their responses challenges the assumption that one or a few genes control permethrin metabolic resistance. Attempts to identify one or a few metabolic genes that are predictably associated with permethrin adaptation may be futile.

QTL mapping of genome regions controlling temephos resistance in larvae of the mosquito Aedes aegypti.

Introduction The mosquito Aedes aegypti is the principal vector of dengue and yellow fever flaviviruses. Temephos is an organophosphate insecticide used globally to suppress Ae. aegypti larval populations but resistance has evolved in many locations. Methodology/Principal Findings Quantitative Trait Loci (QTL) controlling temephos survival in Ae. aegypti larvae were mapped in a pair of F3 advanced intercross lines arising from temephos resistant parents from Solidaridad, México and temephos susceptible parents from Iquitos, Peru. Two sets of 200 F3 larvae were exposed to a discriminating dose of temephos and then dead larvae were collected and preserved for DNA isolation every two hours up to 16 hours. Larvae surviving longer than 16 hours were considered resistant. For QTL mapping, single nucleotide polymorphisms (SNPs) were identified at 23 single copy genes and 26 microsatellite loci of known physical positions in the Ae. aegypti genome. In both reciprocal crosses, Multiple Interval Mapping identified eleven QTL associated with time until death. In the Solidaridad×Iquitos (SLD×Iq) cross twelve were associated with survival but in the reciprocal IqxSLD cross, only six QTL were survival associated. Polymorphisms at acetylcholine esterase (AchE) loci 1 and 2 were not associated with either resistance phenotype suggesting that target site insensitivity is not an organophosphate resistance mechanism in this region of México. Conclusions/Significance Temephos resistance is under the control of many metabolic genes of small effect and dispersed throughout the Ae. aegypti genome.

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

ABSTRACT 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.

Detección de Cocaína en Larvas de Dípteros Necrófagos en Monterrey, Nuevo León, México

Resumen. En el Servicio Médico Forense de la Procuraduría de Justicia del Estado de Nuevo León, se revisaron siete cadáveres y/o fragmentos de humanos, de los cuales se sospechaba la presencia de sustancias tóxicas. Se realizaron estudios toxicológicos para rastrear psicotrópicos, estupefacientes y alcohol etílico. Se recolectaron larvas de Cochliomya macellaria (Fabricius), Chrysomya rufifacies Macquart), Sarcophaga haemorrhoidalis (Fallen), y Musca domestica (Linnaeus). Se detectó cocaína en las larvas obtenidas de cuatro cadáveres, con excepción del sarcofágido, por medio de un Cromatógrafo de Líquidos de Alta Resolución.