Insecticide resistance in Aedes aegypti from Tapachula, Mexico: Spatial variation and response to historical insecticide use.

BACKGROUND: Insecticide use continues as the main strategy to control Aedes aegypti, the vector of dengue, Zika, chikungunya, and yellow fever. In the city of Tapachula, Mexico, mosquito control programs switched from pyrethroids to organophosphates for outdoor spatial spraying in 2013. Additionally, the spraying scheme switched from total coverage to focused control, prioritizing areas with higher entomological-virological risk. Five years after this strategy had been implemented, we evaluated the status and variability of insecticide resistance among Ae. aegypti collected at 26 sites in Tapachula. METHODOLOGY/PRINCIPAL FINDINGS: We determined the lethal concentrations at 50% of the tested populations (LC50) using a bottle bioassay, and then, we calculated the resistance ratio (RR) relative to the susceptible New Orleans strain. Permethrin and deltamethrin (pyrethroids), chlorpyrifos and malathion (organophosphates), and bendiocarb (carbamate) were tested. The frequencies of the substitutions V1016I and F1534C, which are in the voltage-gated sodium channel and confer knockdown-resistance (kdr) to pyrethroid insecticides, were calculated. Despite 5 years having passed since the removal of pyrethroids from the control programs, Ae. aegypti remained highly resistant to permethrin and deltamethrin (RR > 10-fold). In addition, following 5 years of chlorpyrifos use, mosquitoes at 15 of 26 sites showed moderate resistance to chlorpyrifos (5- to 10-fold), and the mosquitoes from one site were highly resistant. All sites had low resistance to malathion (< 5-fold). Resistance to bendiocarb was low at 19 sites, moderate at five, and high at two. Frequencies of the V1016I ranged from 0.16-0.71, while C1534 approached fixation at 23 sites (0.8-1). Resistance profiles and kdr allele frequencies varied across Tapachula. The variability was not associated with a spatial pattern at the scale of the sampling. CONCLUSION/SIGNIFICANCE: Mosquito populations respond to selection pressure at a focal scale in the field. Spatial variation across sites highlights the importance of testing multiple sites within geographical regions.

Composition and abundance of anopheline species according to habitat diversity in Mexico / Composición y abundancia de especies de anofelinos según la diversidad de hábitats en México

Abstract: Objective: To determine the abundance and geographic distribution of the main malaria vectors, which are influenced by habitat characteristics and ecological factors that directly impact adult density and the dynamics of malaria transmission in Mexico. Materials and methods: Samples of larvae were collected from 19 states in Mexico. Each larval habitat was characterized in situ determining the following parameters: water depth, turbidity, percentage of vegetation cover, amount of detritus, presence of algae, light intensity, type of vegetation, amount of predators, habitat stability, altitude, and hydrologic type. Results: A total of 21 687 larvae corresponding to 13 anopheline species were obtained from 149 aquatic habitats. The most abundant species were Anopheles pseudopunctipennis (52.91%), An. albimanus (39.14%) and An. franciscanus (5.29%). The multiple logistic regression analysis showed a negative association between An. pseudopunctipennis and water turbidity (ß=-1.342; Wald=6.122; p=0.013) and the amount of detritus (ß=-2.206; Wald=3.642; p=0.050). While in An. albimanus, there was a significant positive association with water turbidity (ß=1.344; Wald=4.256; p=0.039), a negative correlation was found with the altitude (ß=-3.445; Wald=5.407; p=0.020). The highest mosquito species diversity index was found in Chiapas (Fisher's α=1.20) and the lowest diversity in Chihuahua (Fisher's α=0.26). The greatest richness was found in streams (n=11). Conclusions: The two most abundant species were: An. albimanus and An. pseudopunctipennis. Detailed knowledge of the distribution and characteristics of their larval habitats will be useful for the effective implementation of control strategies in Mexico.

Resumen Objetivo: Determinar la abundancia y la distribución geográfica de los principales vectores de la malaria, las cuales están influenciadas por las características del hábitat y los factores ecológicos que afectan directamente la densidad de los adultos y la dinámica de la transmisión de la malaria en México. Material y métodos: Se obtuvieron muestras de larvas de 19 estados de México. Cada hábitat larvario se caracterizó in situ determinando los siguientes parámetros: profundidad del agua, turbidez, porcentaje de cobertura vegetal, cantidad de detritus, presencia de algas, intensidad de luz, tipo de vegetación, cantidad de depredadores, estabilidad del hábitat, altitud y tipo hidrológico. Resultados: Se identificaron un total de 21 687 larvas pertenecientes a 13 especies de anofelinos, de 149 hábitats acuáticos. Las tres especies más abundantes fueron Anopheles pseudopunctipennis (52.91%), An. albimanus (39.14%) y An. franciscanus (5.29%). El análisis de regresión logística múltiple mostró una asociación negativa para An. pseudopunctipennis y la turbidez del agua (ß=-1.342; Wald= 6.122; p=0.013) y la cantidad de detritus (ß=-2.206; Wald= 3.642; p=0.050). Para An. albimanus se encontró una asociación positiva significativa con la turbidez del agua (ß=1.344; Wald= 4.256; p=0.039) y una correlación negativa con la altitud (ß=-3.445; Wald=5.407; p=0.020). El índice de diversidad más alto se encontró en Chiapas (α de Fisher=1.20) y la diversidad más baja en Chihuahua (α de Fisher=0.26). La mayor riqueza se encontró en los arroyos (n=11). Conclusiones: Las dos especies más abundantes fueron An. albimanus y An. pseudopunctipennis. El conocimiento detallado de la distribución y características de sus hábitats larvales será útil para la implementación efectiva de las estrategias de control en México.

Determination of insecticides' lethal concentrations and metabolic enzyme levels in Triatoma dimidiata / Determinación de concentraciones letales y niveles de enzimas metabólicas de insecticidas en Triatoma dimidiata

Abstract: Objective: The feasibility of the use of WHO impregnated paper and biochemical assays to determine lethal concentrations (LC50 and LC99) and insecticide metabolic enzyme levels of Triatoma dimidiata. Materials and methods: LC50 and LC99 were calculated with WHO papers impregnated at different concentrations of malathion, propoxur and deltamethrin; the percentage of insensitive acetylcholinesterase (iAChE); and the levels of esterases, glutathione S-transferases, and monooxygenases in laboratory nymphs of the first stage (5 to 7 days), were undertaken using the WHO biochemical assays. Results: Respectively the LC50 and LC99 µg/cm2 obtained for malathion were 43.83 and 114.38, propoxur 4.71 and 19.29, and deltamethrin 5.80 and 40.46. A 30% of the population had an iAChE, and only a few individuals had high P450 and β-eterase levels. Conclusion: Impregnated papers and biochemical tests developed by WHO for other insects, proved to be feasible methods in monitoring insecticide resistance and metabolic enzymes involved in T. dimidiata.

Resumen: Objetivo: La factibilidad de usar los papeles impregnados y ensayos bioquímicos según la OMS para determinar concentraciones letales (CL50 y CL99) y niveles enzimáticos en la resistencia a insecticidas en Triatoma dimidiata. Material y métodos: Se calcularon la CL50 y CL99 con papeles impregnados según la OMS a diferentes concentraciones de malatión, propoxur y deltametrina; el porcentaje de acetilcolinesterasa insensible (iAChE); y los niveles de esterasas, glutatión S-transferasas, y monooxigenasas en ninfas de laboratorio del estadio I (5-7 días) se determinaron usando los ensayos bioquímicos según la OMS. Resultados: Se obtuvieron las CL50 y CL99 µg / cm2 respectivamente para malatión 43.83 y 114.38, propoxur 4.71 y 19.29, y deltametrina 5.80 y 40.46. Un 30% de las chinches tuvo iAChE, y sólo pocos individuos tuvieron niveles superiores de P450 y β-eterasas. Conclusión: Los papeles impregnados y ensayos bioquímicos que describe la OMS para otros insectos demostraron ser métodos factibles para monitorear la resistencia a insecticidas y las enzimas metabólicas involucradas en T. dimidiata.

Determination of insecticides' lethal concentrations and metabolic enzyme levels in Triatoma dimidiata.

OBJECTIVE: The feasibility of the use of WHO impregnated paper and biochemical assays to determine lethal concentrations (LC50 and LC99) and insecticide metabolic enzyme levels of Triatoma dimidiata. MATERIALS AND METHODS: LC50 and LC99 were calculated with WHO papers impregnated at different concentrations of malathion, propoxur and deltamethrin; the percentage of insensitive acetylcholinesterase (iAChE); and the levels of esterases, glutathione S-transferases, and monooxygenases in laboratory nymphs of the first stage (5 to 7 days), were undertaken using the WHO biochemical assays. RESULTS: Respectively the LC50 and LC99 µg/cm2 obtained for malathion were 43.83 and 114.38, propoxur 4.71 and 19.29, and deltamethrin 5.80 and 40.46. A 30% of the population had an iAChE, and only a few individuals had high P450 and ß-eterase levels. CONCLUSIONS: Impregnated papers and biochemical tests developed by WHO for other insects, proved to be feasible methods in monitoring insecticide resistance and metabolic enzymes involved in T. dimidiata.

OBJETIVO: La factibilidad de usar los papeles impregnados y ensayos bioquímicos según la OMS para determinar concentraciones letales (CL50 y CL99) y niveles enzimáticos en la resistencia a insecticidas en Triatoma dimidiata. MATERIAL Y MÉTODOS: Se calcularon la CL50 y CL99 con papeles impregnados según la OMS a diferentes concentraciones de malatión, propoxur y deltametrina; el porcentaje de acetilcolinesterasa insensible (iAChE); y los niveles de esterasas, glutatión S-transferasas, y monooxigenasas en ninfas de laboratorio del estadio I (5-7 días) se determinaron usando los ensayos bioquímicos según la OMS. RESULTADOS: Se obtuvieron las CL50 y CL99 µg / cm2 respectivamente para malatión 43.83 y 114.38, propoxur 4.71 y 19.29, y deltametrina 5.80 y 40.46. Un 30% de las chinches tuvo iAChE, y sólo pocos individuos tuvieron niveles superiores de P450 y ß-eterasas. CONCLUSIONES: Los papeles impregnados y ensayos bioquímicos que describe la OMS para otros insectos demostraron ser métodos factibles para monitorear la resistencia a insecticidas y las enzimas metabólicas involucradas en T. dimidiata.

Composition and abundance of anopheline species according to habitat diversity in Mexico.

OBJECTIVE: To determine the abundance and geographic distribution of the main malaria vectors, which are influenced by habitat characteristics and ecological factors that directly impact adult density and the dynamics of malaria transmission in Mexico. MATERIALS AND METHODS: Samples of larvae were collected from 19 states in Mexico. Each larval habitat was characterized in situ determining the following parameters: water depth, turbidity, percentage of vegetation cover, amount of detritus, presence of algae, light intensity, type of vegetation, amount of predators, habitat stability, altitude, and hydrologic type. RESULTS: A total of 21 687 larvae corresponding to 13 anopheline species were obtained from 149 aquatic habitats. The most abundant species were Anopheles pseudopunctipennis (52.91%), An. albimanus (39.14%) and An. franciscanus (5.29%). The multiple logistic regression analysis showed a negative association between An. pseudopunctipennis and water turbidity (ß=-1.342; Wald=6.122; p=0.013) and the amount of detritus (ß=-2.206; Wald=3.642; p=0.050). While in An. albimanus, there was a significant positive association with water turbidity (ß=1.344; Wald=4.256; p=0.039), a negative correlation was found with the altitude (ß=-3.445; Wald=5.407; p =0.020). The highest mosquito species diversity index was found in Chiapas (Fisher's α=1.20) and the lowest diversity in Chihuahua (Fisher's α=0.26). The greatest richness was found in streams (n=11). CONCLUSIONS: The two most abundant species were: An. albimanus and An. pseudopunctipennis. Detailed knowledge of the distribution and characteristics of their larval habitats will be useful for the effective implementation of control strategies in Mexico.

OBJETIVO: Determinar la abundancia y la distribución geográfica de los principales vectores de la malaria, las cuales están influenciadas por las características del hábitat y los factores ecológicos que afectan directamente la densidad de los adultos y la dinámica de la transmisión de la malaria en México. MATERIAL Y MÉTODOS: Se obtuvieron muestras de larvas de 19 estados de México. Cada hábitat larvario se caracterizó in situ determinando los siguientes parámetros: profundidad del agua, turbidez, porcentaje de cobertura vegetal, cantidad de detritus, presencia de algas, intensidad de luz, tipo de vegetación, cantidad de depredadores, estabilidad del hábitat, altitud y tipo hidrológico. RESULTADOS: Se identificaron un total de 21 687 larvas pertenecientes a 13 especies de anofelinos, de 149 hábitats acuáticos. Las tres especies más abundantes fueron Anopheles pseudopunctipennis (52.91%), An. albimanus (39.14%) y An. franciscanus (5.29%). El análisis de regresión logística múltiple mostró una asociación negativa para An. pseudopunctipennis y la turbidez del agua (ß=-1.342; Wald= 6.122; p=0.013) y la cantidad de detritus (ß=-2.206; Wald= 3.642; p=0.050). Para An. albimanus se encontró una asociación positiva significativa con la turbidez del agua (ß=1.344; Wald= 4.256; p=0.039) y una correlación negativa con la altitud (ß=-3.445; Wald=5.407; p=0.020). El índice de diversidad más alto se encontró en Chiapas (α de Fisher=1.20) y la diversidad más baja en Chihuahua (α de Fisher=0.26). La mayor riqueza se encontró en los arroyos (n=11). CONCLUSIONES: Las dos especies más abundantes fueron An. albimanus y An. pseudopunctipennis. El conocimiento detallado de la distribución y características de sus hábitats larvales será útil para la implementación efectiva de las estrategias de control en México.

Larval habitat characterization of Anopheles darlingi from its northernmost geographical distribution in Chiapas, Mexico.

BACKGROUND: Anopheles darlingi is considered the most efficient malaria vector in the Neotropical region. In Mexico, its role as an incriminated vector of Plasmodium has not been confirmed in the Lacandon forest. Similarly, knowledge about bionomic and larval ecology is scarce. The study aim was to identify and describe the larval habitats of An. darlingi in Chiapas, México. METHODS: Standard larval collections were performed in the Lacandon forest region and in the Soconusco region of southern Chiapas from January 2010 to April 2014, including dry and rainy seasons. Mean larval density of An. darlingi was estimated according to hydrological types, and associations between the presence of An. darlingi and environmental factors including ecological parameters and geographic positions were statistically analysed. RESULTS: One hundred and twelve aquatic habitats were analysed, 80 from the Lacandon forest region and 32 from the Soconusco region; 94.64% of these sites presented anopheline larvae. In total, 10,977 larvae belonging to 11 Anopheles species were collected. The 19 (out of 112) larval habitats positive to An. darlingi were: rain puddles (26.32%), ground pools (21.05%), ponds (15.79%), ditches (15.79%), river margins (10.53%) and streams (10.53%). Overall, the average (±SD) larval density was 6.60 ± 2.41 larvae per dip. Multiple logistic regression analysis showed that temporary habitats, green algae presence and stagnant water were associated with An. darlingi larval presence. The positive habitats were found in the Lacandon forest region during the rainy season (May-September). No specimens were found in the Soconusco region of the coastal plain of Chiapas. CONCLUSION: The mosquito An. darlingi larval habitats were found in different hydrological types. The habitat stability, presence of algae and water current were the main factors for An. darlingi larval occurrence. The information on the characteristics of the larval habitats of An. darlingi will be useful in sustainable programmes for malaria control in the Lacandon forest region, Chiapas.