Pyrethroids are commonly used as mosquito adulticides and evolution of resistance to these compounds is a major threat to public health. 'Knockdown resistance' to pyrethroids (kdr) is frequently caused by nonsynonymous mutations in the voltage-gated sodium channel transmembrane protein (para) that reduce pyrethroid binding. Early detection of kdr is critical to the development of resistance management strategies in mosquitoes including Aedes aegypti, the most prevalent vector of dengue and yellow fever viruses. Brengues et al. described seven novel mutations in hydrophobic segment 6 of domain II of para in Ae. aegypti. Assays on larvae from strains bearing these mutations indicated reduced nerve sensitivity to permethrin inhibition. Two of these occurred in codons Iso1011 and Val1016 in exons 20 and 21 respectively. A transition in the third position of Iso1011 encoded a Met1011 replacement and a transversion in the second position of Val1016 encoded a Gly1016 replacement. We have screened this same region in 1318 mosquitoes in 32 additional strains; 30 from throughout Latin America. While the Gly1016 allele was never detected in Latin America, we found two new mutations in these same codons. A transition in the first position of codon 1011 encodes a Val replacement while a transition in the first position of codon 1016 encodes an Iso replacement. We developed PCR assays for these four mutations that can be read either on an agarose gel or as a melting curve. Selection experiments, one with deltamethrin on a field strain from Santiago de Cuba and another with permethrin on a strain from Isla Mujeres, Mexico rapidly increased the frequency of the Iso1016 allele. Bioassays of F(3) offspring arising from permethrin susceptible Val1016 homozygous parents and permethrin resistant Iso1016 homozygous parents show that Iso1016 segregates as a recessive allele in conferring kdr. Analysis of segregation between alleles at the 1011 and 1016 codons in the F(3) showed a high rate of recombination even though the two codons are only separated by a ~250 bp intron. The tools and information presented provide a means for early detection and characterization of kdr that is critical to the development of strategies for resistance management.

Entomologic indices can identify areas at high risk for disease transmission.

ika virus (ZIKV) is an arthropod-borne flavivirus related to yellow fever, dengue and West Nile viruses 1. ZIKV was first reported in East Africa in 1947 and expanded from the ancestral enzootic cycle in Africa to Asia several decades ago (Fig. 1). At the beginning of the 21st century, the virus expanded into the South Pacific and the Americas, triggering a pandemic that led to 48 countries reporting active ZIKV transmission by 2017 2. Prior to this expansion, there was little scientific research on this virus 3. In this paper, we review the natural history of ZIKV and the current knowledge about ZIKV vector-borne transmission and the mosquito and vertebrate host species potentially involved worldwide. Furthermore, we discuss the possibility of ZIKV spillback into an enzootic cycle outside Africa and review hypotheses regarding ZIKV recent global emergence and evolution. Finally, we identify research priorities for filling remaining gaps and challenges in our understanding of ZIKV. Zika virus natural history The virus was first isolated from a sentinel rhesus macaque and from Aedes (Stegomyia) africanus mosquitoes in the same Ugandan location. Surveillance efforts identified immune people in at least 25 African countries from 1945 to 2014 (reviewed in ref. 4) and in 7 Asian countries or territories from 1952 to 1997 (Fig. 1). However, many of the serologic tests employed in this surveillance are cross-reactive among flaviviruses, thus these results must be interpreted with caution. Nevertheless, direct detection of ZIKV in countries including Senegal,

Eight Latin American strains of Aedes aegypti were evaluated for resistance to 6 organophosphates (temephos, malathion, fenthion, pirimiphos-methyl, fenitrothion, and chlorpirifos) and 4 pyrethroids (deltamethrin, lambdacyhalothrin, betacypermethrin, and cyfluthrin) under laboratory conditions. In larval bioassays, temephos resistance was high (resistance ratio [RR50], > or =10X) in the majority of the strains, except for the Nicaragua and Venezuela strains, which showed moderate resistance (RR50, between 5 and 10X). The majority of the strains were susceptible to malathion, fenthion, and fenitrothion. However, resistance to pirimiphos-methyl ranged from moderate to high in most of the strains. Larvae from Havana City were resistant to 3 of the pyrethroids tested and moderately resistant to cyfluthrin. The Santiago de Cuba strain showed high resistance to deltamethrin and moderate resistance to the other pyrethroids (lambdacyhalothrin, betacypermethrin, and cyfluthrin). The rest of the strains were susceptible to pyrethroids, except for the Jamaica and Costa Rica strains, which showed moderate resistance to cyfluthrin, and Peru and Venezuela, which showed resistance to deltamethrin. Adult bioassays showed that all the strains were resistant to dichlorodiphenyl-trichloroethane and to the majority of pyrethroids evaluated. The use of the synergists S,S,S,-tributyl phosphorotrithioate and piperonil butoxide showed that esterase and monooxygenases played an important role in the temephos, pirimiphos-methyl, and chlorpirifos resistance in some strains. Biochemical tests showed high frequencies of esterase and glutathione-S-transferase activity; however, the frequency of altered acetylcholinesterase mechanism was low. The polyacrylamide electrophoresis gel detected the presence of a strong band called Est-A4. Insecticide resistance in Ae. aegypti is a serious problem facing control operations, and integrated control strategies are recommended to help prevent or delay the temephos resistance in larvae and pyrethroids resistance in adults.

A sample of Aedes aegypti (L.) from Santiago de Cuba, Cuba, with a high level of temephos resistance (19.58x at the 50% lethal concentration [LC50]), was subjected to temephos selection to evaluate the utility of this organophosphate insecticide for mosquito control. High resistance developed after six generations of selection (200.00x). Little or no cross-resistance was observed to the organophosphates, malathion and fenitrothion, but high cross-resistance was observed for the pyrethroid deltamethrin (337.5x) and the organophosphate fenthion (12.74x). Synergism tests implicated detoxifying esterases in temephos and fenthion resistance and deltamethrin resistance was associated with a cytochrome P450 monooxygenasa. Biochemical tests, polyacrylamide gel electrophoresis (PAGE), and inhibition studies confirmed the presence of elevated esterase activity associated with temephos resistance. Pyrethroid resistance could be associated with a glutathione-S-transferase mechanism but not the esterases. The cross-resistance to deltamethrin from temephos selection could limit the use of both insecticides for Ae. aegypti control.