Aedes aegypti (L.) is the primary vector of the viruses that cause dengue, Zika, and chikungunya, for which effective vaccines and drugs are still lacking. Current strategies for suppressing arbovirus outbreaks based on insecticide use pose a challenge because of the rapid increase in resistance. The widespread and excessive use of pyrethroid-based insecticides has created a large selection pressure for a kdr-type resistance, caused by mutations in the para gene of the voltage-gated sodium channel (vgsc). Our objective was to evaluate the allelic frequency of natural populations of Ae. aegypti of Mexico at codon 410 of the para gene. Twenty-six Ae. aegypti populations from east and southern Mexico were genotyped for the codon 410 using allele-specific PCR. The frequencies of the L410 allele in Ae. aegypti ranged from 0.10 to 0.99; however, most of the frequencies were in the range of 0.36 to 0.64. The highest frequencies were found in three populations from the state of Veracruz, namely, Minatitlan with 0.99, Poza Rica with 0.82, and Jose Cardel with 0.97, along with populations from Cancun in Quintana Roo with 0.93, Frontera in Tabasco with 0.91, and Ciudad del Carmen in Campeche with 0.86. The frequency of the L410 allele was high in all populations of Ae. aegypti with higher values in populations of the southeast of the country. The knowledge of specific substitutions in vgsc and their interaction to confer resistance is essential to predict and develop future strategies for resistance management in Ae. aegypti in Mexico.
Background: Knockdown resistance (kdr) is the main mechanism that confers resistance to pyrethroids and DDT. This is a product of non-synonymous mutations in the voltage-gated sodium channel (vgsc) gene, and these mutations produce a change of a single amino acid which reduces the affinity of the target site for the insecticide molecule. In Mexico, V410L, V1016I and F1534C mutations are common in pyrethroid-resistant Aedes aegypti (L.) populations. Methods: A multiplex PCR was developed to detect the V410L, V1016I and F1534C mutations in Ae. aegypti. The validation of the technique was carried out by DNA sequencing using field populations previously characterized for the three mutations through allele-specific PCR (AS-PCR) and with different levels of genotypic frequencies. Results: The standardized protocol for multiplex end-point PCR was highly effective in detecting 15 genotypes considering the three mutations V410L, V1106I and F1534C, in 12 field populations of Ae. aegypti from Mexico. A complete concordance with AS-PCR and DNA sequencing was found for the simultaneous detection of the three kdr mutations. Conclusions: Our diagnostic method is highly effective for the simultaneous detection of V410L, V1016I and F1534C, when they co-occur. This technique represents a viable alternative to complement and strengthen current monitoring and resistance management strategies against Ae. aegypti.
Although having five different ways of transmission the vector-borne is the principal way of transmission of Chagas disease, which involves insects of the subfamily Triatominae (Hemiptera: Reduviidae). Nineteen of the 31 species that occur in Mexico are associated with humans, and all are capable of transmitting the disease. Pyrethroids are the insecticides recommended for the control of these vectors in Mexico. We determined the susceptibility to the pyrethroids deltamethrin and permethrin of peridomestic populations of Triatoma mazzottii Usinger and two populations of Triatoma longipennis Usinger in comparison with a reference strain for each species. Bioassays were performed for the determination of the LD for both field populations and reference strains. A maximum of 27 fold resistance to deltamethrin was observed in T. mazzottii, meanwhile, for permethrin, T. longipennis from Jalisco show the highest value of 3.19 fold. There was significantly increased activity of esterases in field populations in comparison with their corresponding reference strain. The results of the search of kdr mutations related to the resistance to deltamethrin and permethrin in the evaluated species show the presence of mutations in the field populations, as is the case with individuals of T. mazzottii, for which the mutation was found A943V, and for the two populations of T. longipennis included in this study, we report the presence of the kdr mutation K964R. Evaluation of the various mechanisms involved in resistance to pyrethroids in triatomines from Mexico could guide us to the real justification for insecticide resistance monitoring.
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