Determinación in vivo del papel de las enzimas esterasas y glutation transferasa en la resistencia a piretroides en Aedes aegypti (Diptera: Culicidae) / Determination in vivo of the role of esterase and gluthation transferase enzymes in pyrethroid resistance of Aedes aegypti (Diptera: Cullicidae)

Se realizó un estudio in vivo a través del uso de 2 sinergistas, el trifenil fosfato (TFF) inhibidor específico de esterasas y el ácido etacrínico (AE), inhibidor específico de la enzima glutation transferasa (GST), para determinar si estas enzimas eran responsables de la resistencia a piretroides en Aedes aegypti. Para el trabajo se utilizaron 2 cepas de Aedes aegypti resistentes a insecticidas, una cepa que fue seleccionada con temefos por 6 generaciones de selección (SAN-F6) y otra con deltametrina por 12 generaciones de selección con este insecticida (SAN-F12), ambas resultaron ser resistentes a insecticidas piretroides. Se demostró a través del uso de los sinergistas TFF y AE que las enzimas esterasas y GST son responsables de la resistencia a los piretroides en estas cepas. Esos resultados demuestran la existencia de un fenómeno de resistencia cruzada y multirresistencia, lo cual debe tenerse en cuenta en las estrategias de uso de insecticidas para el control de este vector.

An in vivo study of two synergists, that is, Triphenil phosphate -specific esterase inhibitor- and ethacrynic acid specific gluthation transferase inhibitor- was performed to determine if these enzymes were responsible for pyrethroid resistance of Aedes aegypti. To this end, two insecticide resistant Aedes aegypti strains were used, one strain selected with temephos by six selection generations (SAN-F6) and the other strain with delmamethrin by 12 selection generations (SAN-F12), being both strains resistant to pyrethroid insecticices. Through the use of TPP and EA synergists, it was proved that esterase and gluthation-s-transferase (GST) enzymes were responsible for pryrethroid resistance of these strains. These results showed the existence of cross-resistance and multidrug resistance, which should be taken into account for insecticide use strategies aimed at vector control.

[Insecticide resistance mechanisms of Aedes aegypti (Diptera: Culicidae) from two Peruvian provinces]. / Resistencia a insecticidas y mecanismos de resistencia en Aedes aegypti (Diptera: Culicidae) de 2 provincias del Perú.

Insecticide resistance of Aedes aegypti larvae and adults from two Peruvian provinces, that is, Trujillo and Tumbes provinces, was conducted. High infestation indexes and extensive use of insecticides based on the Vector Surveillance and Control Strategy of the Ministry of Public Health prevailed in these places. Larval bioassays revealed susceptibility to organophosphorate insecticide called malathion in TRUJILLO strain, it being moderate to fention and fenitrotion and high to chlorpyriphos and temephos;however, TUMBES strain was susceptible to the evaluated organophosphorate compounds, except for fention, with moderate resistance. In the adult state, at the recommended dose, TRUJILLO strain showed resistence to DDT organochlorate insecticide and to pyrethoids called lambdacyalotrine and cyflutrine whereas TUMBES was resistant to DDT and to all assessed pyrethoids. None of them was resistant to chlorpiriphos in adult stage. By using synergists, the results showed that esterases and monooxigenases played an important role in the detected resistence to organophosphorate in Aedes larvae from TRUJILLO province. Biochemical assays yielded that increased activity of esterases was very frequent in TRUJILLO strain as was the case of glutathion transferase(GST) and modified acetylcholinesterase (AchR). On the other hand, the polyacrylamide gel electrophoresis allowed observing the prevalence of amplified activity of esterases A4 in TRUJILLO strain but not in TUMBES strain.

[Determination in vivo of the role of esterase and glutathione transferase enzymes in pyrethroid resistance of Aedes aegypti (Diptera: Culicidae)]. / Determinación in vivo del papel de las enzimas esterasas y glutation transferasa en la resistencia a piretroides en Aedes aegypti (Diptera: Culicidae).

An in vivo study of two synergists, that is, Triphenil phosphate -specific esterase inhibitor- and ethacrynic acid -specific gluthation transferase inhibitor- was performed to determine if these enzymes were responsible for pyrethroid resistance of Aedes aegypti. To this end, two insecticide resistant Aedes aegypti strains were used, one strain selected with temephos by six selection generations (SAN-F6) and the other strain with delmamethrin by 12 selection generations (SAN-F12), being both strains resistant to pyrethroid insecticices. Through the use of TPP and EA synergists, it was proved that esterase and gluthation-s-transferase (GST) enzymes were responsible for pryrethroid resistance of these strains. These results showed the existence of cross-resistance and multidrug resistance, which should be taken into account for insecticide use strategies aimed at vector control.

Levels of insecticide resistance and resistance mechanisms in Aedes aegypti from some Latin American countries.

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.

Selection of insensitive acetylcholinesterase as a resistance mechanism in Aedes aegypti (Diptera: Culicidae) from Santiago de Cuba.

A sample of Aedes aegypti (L.) (Diptera: Culicidae) from Santiago de Cuba, Cuba, with a high level of propoxur resistance compared with the reference susceptible Rockefeller strain (12.60 x at the 50% lethal concentration [LC50] and 18.08 at the 90% lethal concentration [LC90]), with a 4.3% frequency of insensitive acetylcholinesterase (AChE) frequency, was subjected to propoxur selection for 13 successive generations to increase the frequency of this resistance mechanism in Ae. aegypti. High resistance to propoxur was developed during this selection (41.73-fold), and the frequency of insensitive AChE mechanism was increased 13.25-fold. Other mechanisms (overproduced esterases, glutathione transferases, or monooxygenases) were not detected in the propoxur-selected strain. The selection of an insensitive AChE resistance mechanism in Ae. aegypti has important implications and will be a valuable resource for genetic studies and molecular characterization of the ace gene mutation(s) associated with insecticide resistance in Ae. aegypti.

Resistencia a insecticidas en larvas y adultos de Aedes aegypti: prevalencia de la esterasa A4 asociada con la resistencia a temefos / Resistance to insecticides in larvae and adults of Aedes aegypti: prevalence of A4 esterasa associated with resistance to temephos

Fueron estudiadas 2 cepas de Aedes aegypti provenientes de 2 Consejos populares con altos índices de infestación de este vector, producto de la necesidad de hacer un control efectivo de larvas y adultos en el municipio Guanabacoa. Se determinaron los niveles de susceptibilidad y/o resistencia a insecticidas organofosforados, piretroides y un carbamato. Los resultados de los bioensayos en larvas mostraron completa susceptibilidad a los insecticidas organofosforados malation, clorpirifos, pirimifos metil y al carbamato propoxur en ambos consejos populares, sin embargo, se observó alta resistencia a temefos y a fention, no se observó resistencia a fenitrotion en uno de los consejos populares, pero sí en el otro. En el estado adulto, a la dosis recomendada por la Organización Mundial de la Salud o por los fabricantes de los productos, se observó resistencia a malation, fenitrotion y propoxur, sin embargo, se obtuvieron mejores resultados con los piretroides, con porcentajes de mortalidad por encima de 90 por ciento. Según los resultados obtenidos, utilizando el sinergista S S S tributil fosforotritiado (DEF) y piperonil butóxido, se demostró que las enzimas esterasas y oxidasas de función múltiple desempeñaron un papel importante en la resistencia a temefos y a fention. Por medio de ensayos bioquímicos se demostró que la amplificada actividad de estas enzimas se encontró a elevada frecuencia en ambos consejos populares, al igual que la enzima glutation transferasa (GST) en uno de los 2 Consejos Populares. Mediante electroforesis en gel de poliacrilamida se observó que en ambos consejos populares prevaleció la amplificada actividad de esterasas A4

Estado de la resistencia a insecticidas y mecanismos de resistencia en larvas del municipio Playa, colectadas durante la etapa intensiva contra el Aedes aegypti en Ciudad de La Habana, 2001-2002 / State of the resistance to insecticides and resistance mechanisms in larvae from Playa municipality collected during the intensive campaign against Aedes aegypti in Havana City, 2001-2002

Se determinaron los niveles de susceptibilidad y/o resistencia a insecticidas en larvas de Aedes aegypti, colectadas del municipio Playa, durante la etapa intensiva de la campaña contra el mosquito Aedes aegypti en Ciudad de La Habana, en el presente año. En larvas, se encontró alta resistencia a los insecticidas organofosforados fention y temefos y moderada a fenitrotion, también se detectó alta resistencia al carbamato propoxur. Las larvas resultaron susceptibles a clorpirifos, malation y pirimifos metil. Los resultados a través del uso de sinergistas indicaron que las esterasas desempeñan un papel importante en la resistencia detectada a los insecticidas organofosforados, pero no en la resistencia a propoxur, sin embargo, las monoxigenasas intervienen solamente en la resistencia a fenitrotion. Estos mecanismos se encontraron a elevada frecuencia en las larvas de este municipio. Se confirmó el papel de las esterasas en la resistencia a temefos mediante ensayos de inhibición en gel de poliacrilamida

[Resistance to insecticides in larvae and adults of Aedes aegypti, Havana City: prevalence of A4 esterasa associated with resistance to temephos]. / Resistencia a insecticidas en larvas y adultos de Aedes aegypti: prevalencia de la esterasa A4 asociada con la resistencia a temefos.

2 strains of Aedes aegypti from 2 people's councils with high indexes of infestation of this vector were studied due to the need of carrying out an effective control of larvae and adults in the municipality of Guanabacoa. The levels of susceptibility and/or resistance to organophosphate insecticides, pyrethroids and a carbamate were determined. The results of the bioassays in larvae showed a complete susceptibility to organophosphate insecticides, malathion, clorpirifos, methyl-pyrimifos and propoxur carbamate in both people's councils. However, it was observed a high resistance to temephos and fenthion. No resistance to fenitrothion was found in one of the people's councils. At the adult stage and at the dose recommended by the World Health Organizaton or by the product manufacturers, it was observed resistance to malathion, fenitrothion and propoxur; nevertheless, better results were obtained with the pyrethroids, with mortality percentages over 90%. According to the results obtained, by using the SSS tributyl phosphotritiade sinergist (DEF) and piperonyl butoxide, it was demonstrated that the multiple function esterases and oxidases played an important role in the resistance to temephos and fenthion. It was proved that he amplified activity of these enzymes was at an elevated frequency in both people's councils, the same as the glutathione transferase (GST) in one of the 2 people's councils. It was observed that in both people's councils prevailed the amplified activity of A4 esterases by polyacrylamide gel electrophoresis.

[Status of resistance to insecticides and resistance mechanisms in larvae from Playa municipality collected during the intensive campaign against Aedes aegypti in Havana City, 2001-2002]. / Estado de la resistencia a insecticidas y mecanismos de resistencia en larvas del municipio Playa, colectadas durante la etapa intensiva contra el Aedes aegypti en ciudad de la habana, 2001-2002.

The levels of susceptibility and/or resistance to inseciticides in Aedes aegypti larvae collected at the municipality of Playa during the intensive campaign against the mosquito Aedes aegypti, in Havana City, in the present year, were determined. In larvae, it was found a high resistance to the organophosphate insecticides phention and temephos and moderate to fenitrothion. A high resistance to propoxur carbamate was also detected. The larvae proved to be susceptible to clorpirifos, malathion and methylpyrimifos. The results obtained through the use of sinergists indicated that esterases play an important role in the detected resistance to organophosphate insecticides, but not in the resistance to propoxur. However, the monooxigenases take part only in the resistance to fenitothrion. These mechanisms were found at an elevated frequency in the larvae from this municipality. It was confirmed the role played by esterases in the resistance to temephos by polyacrilamide gel inhibition tests.