Insecticide resistance levels and associated mechanisms in three Aedes aegypti populations from Venezuela.

BACKGROUND: The massive use of insecticides in public health has exerted selective pressure resulting in the development of resistance in Aedes aegypti to different insecticides in Venezuela. Between 2010 and 2020, the only insecticides available for vector control were the organophosphates (Ops) fenitrothion and temephos which were focally applied. OBJECTIVES: To determine the state of insecticide resistance and to identify the possible biochemical and molecular mechanisms involved in three populations of Ae. aegypti from Venezuela. METHODS: CDC bottle bioassays were conducted on Ae. aegypti collected between October 2019 and February 2020 in two hyperendemic localities for dengue in Aragua State and in a malaria endemic area in Bolívar State. Insecticide resistance mechanisms were studied using biochemical assays and polymerase chain reaction (PCR) to detect kdr mutations. FINDINGS: Bioassays showed contrasting results among populations; Las Brisas was resistant to malathion, permethrin and deltamethrin, Urbanización 19 de Abril was resistant to permethrin and Nacupay to malathion. All populations showed significantly higher activity of mixed function oxidases and glutathione-S-transferases (GSTs) in comparison with the susceptible strain. The kdr mutations V410L, F1534C, and V1016I were detected in all populations, with F1534C at higher frequencies. MAIN CONCLUSION: Insecticide resistance persists in three Ae. aegypti populations from Venezuela even in the relative absence of insecticide application.

Malaria entomological risk factors in relation to land cover in the Lower Caura River Basin, Venezuela.

To explore the effects of deforestation and resulting differences in vegetation and land cover on entomological parameters, such as anopheline species composition, abundance, biting rate, parity and entomological inoculation rate (EIR), three villages were selected in the Lower Caura River Basin, state of Bolívar, Venezuela. All-night mosquito collections were conducted between March 2008-January 2009 using CDC light traps and Mosquito Magnet® Liberty Plus. Human landing catches were performed between 06:00 pm-10:00 pm, when anophelines were most active. Four types of vegetation were identified. The Annual Parasite Index was not correlated with the type of vegetation. The least abundantly forested village had the highest anopheline abundance, biting rate and species diversity. Anopheles darlingi and Anopheles nuneztovari were the most abundant species and were collected in all three villages. Both species showed unique biting cycles. The more abundantly forested village of El Palmar reported the highest EIR. The results confirmed previous observations that the impacts of deforestation and resulting changes in vegetation cover on malaria transmission are complex and vary locally.

Can Mosquito Magnet® substitute for human-landing catches to sample anopheline populations?

The efficiency of the Mosquito Magnet Liberty Plus™ (MMLP) trap was evaluated in comparison to human-landing catches (HLCs) to sample anopheline populations in Jabillal, state of Bolivar, southern Venezuela. The village comprised 37 houses and a population of 101; malaria in this village is primarily due to Plasmodium vivax and the Annual Parasite Index is 316.8 per 1,000 population. A longitudinal study was conducted between June 2008-January 2009 for three nights per month every two months between 17:30 pm-21:30 pm, a time when biting mosquitoes are most active. Anopheles darlingi and Anopheles nuneztovari were the most common species collected by both methods, whereas Anopheles marajoara was more abundant according to the HLC method. The MMLP trap was more efficient for collecting An. nuneztovari [63%, confidence interval (CI): 2.53] than for collecting An. darlingi (31%, CI: 1.57). There were significant correlations (p < 0.01) between the two methods for An. darlingi [Pearson correlation (R²) = 0.65] and An. nuneztovari (R² = 0.48). These preliminary results are encouraging for further investigations of the use of the MMLP trap for monitoring anopheline populations in remote malaria-endemic areas in the Amazon Basin.

Mosquito vector biology and control in Latin America--a 22nd symposium.

The 22nd Annual Latin American Symposium presented by the American Mosquito Control Association (AMCA) was held as part of the 78th Annual Meeting in Austin, TX, in February 2012. The principal objective, as for the previous 21 symposia, was to promote participation in the AMCA by vector control specialists, public health workers, and academicians from Latin America. This publication includes summaries of 21 presentations that were given orally in Spanish or presented as posters by participants from Mexico, Colombia, Venezuela, and the USA. Topics addressed in the symposium included surveillance, chemical control, insecticide resistance, and genetics associated with Aedes aegypti; food sources and control of Culex; taxonomy, surveillance, and control of Anopheles vectors of malaria; and studies of dengue virus and Leishmania.

Updating the bionomy and geographical distribution of Anopheles (Nyssorhynchus) albitarsis F: A vector of malaria parasites in northern South America.

Anopheles albitarsis F is a putative species belonging to the Albitarsis Complex, recognized by rDNA, mtDNA, partial white gene, and microsatellites sequences. It has been reported from the island of Trinidad, Venezuela and Colombia, and incriminated as a vector of malaria parasites in the latter. This study examined mitochondrially encoded cytochrome c oxidase I (MT-CO1) sequences of An. albitarsis F from malaria-endemic areas in Colombia and Venezuela to understand its relations with other members of the Complex, revised and update the geographical distribution and bionomics of An. albitarsis F and explore hypotheses to explain its phylogenetic relationships and geographical expansion. Forty-five MT-CO1 sequences obtained in this study were analyzed to estimate genetic diversity and possible evolutionary relationships. Sequences generated 37 haplotypes clustered in a group where the genetic divergence of Venezuelan populations did not exceed 1.6% with respect to Colombian samples. Anopheles albitarsis F (π = 0.013) represented the most recent cluster located closer to An. albitarsis I (π = 0.009). Barcode gap was detected according to Albitarsis Complex lineages previously reported (threshold 0.014-0.021). Anopheles albitarsis F has a wide distribution in northern South America and might play an important role in the transmission dynamics of malaria due to its high expansion capacity. Future studies are required to establish the southern distribution of An. albitarsis F in Venezuela, and its occurrence in Guyana and Ecuador.

Challenges in the control of neglected insect vector diseases of human importance in the Anglo-Caribbean.

INTRODUCTION: Neglected tropical diseases (NTDs) in developing countries like the Caribbean, negatively affect multiple income-generating sectors, including the tourism industry upon which island states are highly dependent. Insect-transmitted NTDs include, but are not limited to, malaria, dengue and lymphatic filariasis. Control measures for these disease, are often ignored because of the associated cost. Many of the developing country members are thus retained in a financially crippling cycle, balancing the cost of prophylactic measures with that of controlling an outbreak.The purpose of the paper is to bring awareness to NTDs transmitted by insects of importance to humans, and to assess factors affecting such control, in the English-speaking Caribbean. METHOD: Comprehensive literature review on reports pertaining to NTDs transmitted by insects in the Caribbean and Latin America was conducted. Data search was carried out on PubMed, and WHO and PAHO websites. RESULTS AND CONCLUSION: Potential risk factors for NTDs transmitted by arthropods in the English-speaking Caribbean are summarised. The mosquito appears to be the main insect-vector of human importance within the region of concern. Arthropod-vectors of diseases of veterinary importance are also relevant because they affect the livelihood of farmers, in highly agriculture based economies. Other NTDs may also be in circulation gauged by the presence of antibodies in Caribbean individuals. However, routine diagnostic tests for specific diseases are expensive and tests may not be conducted when diseases are not prevalent in the population. It appears that only a few English-speaking Caribbean countries have examined secondary reservoirs of pathogens or assessed the effectivity of their insect control methods. As such, disease risk assessment appears incomplete. Although continuous control is financially demanding, an integrated and multisectoral approach might help to deflect the cost. Such interventions are now being promoted by health agencies within the region and various countries are creating and exploring the use of novel tools to be incorporated in their insect-vector control programmes.

Mosquito vector biology and control in Latin America--a 20th symposium.

The 20th Annual Latin American Symposium presented by the American Mosquito Control Association (AMCA) was held as part of the 76th Annual Meeting in Lexington, KY, in March 2010. The principal objective, as for the previous 19 symposia, was to promote participation in the AMCA by vector control specialists, public health workers, and academicians from Latin America. This publication includes summaries of 40 presentations that were given orally in Spanish or presented as posters by participants from 5 countries in Latin America, the United Kingdom, and the USA. Topics addressed in the symposium included: surveillance, chemical and biological control, and insecticide resistance associated with Aedes aegypti; distribution, behavior, and control of Culex; bionomics, ecology, and chemical and biological control of Anopheles vectors of malaria; insecticide resistance; and studies of dengue, West Nile virus, and Triatoma.

Malaria entomological inoculation rates in gold mining areas of Southern Venezuela.

A longitudinal study of malaria vectors aiming to describe the intensity of transmission was carried out in five villages of Southern Venezuela between January 1999-April 2000. The man-biting, sporozoite and entomological inoculation rates (EIR) were calculated based on 121 all-night collections of anophelines landing on humans, CDC light traps and ultra violet up-draft traps. A total of 6,027 female mosquitoes representing seven species were collected. The most abundant species were Anopheles marajoara Galvão & Damasceno (56.7%) and Anopheles darlingi Root (33%), which together accounted for 89.7% of the total anophelines collected. The mean biting rate for An. marajoara was 1.27 (SD + 0.81); it was 0.74 (SD + 0.91) for An. darlingi and 0.11 (SD + 0.10) for Anopheles neomaculipalpus Curry and the overall biting rate was 2.29 (SD + 1.06). A total of 5,886 mosquitoes collected by all three methods were assayed by ELISA and 28 pools, equivalent to 28 mosquitoes, yielded positive results for Plasmodium spp. CS protein. An. neomaculipalpus had the highest sporozoite rate 0.84% (3/356), followed by An. darlingi 0.82% (16/1,948) and An. marajoara 0.27% (9/3,332). The overall sporozoite rate was 0.48% (28/5,886). The rates of infection by Plasmodium species in mosquitoes were 0.37% (22/5,886) for Plasmodium vivax(Grassi & Feletti) and 0.10% (6/5,886) for Plasmodium falciparum (Welch). The estimated overall EIR for An. darlingi was 2.21 infective bites/person/year, 1.25 for An. marajoara and 0.34 for An. neomaculipalpus. The overall EIR was four infective bites/person/year. The biting rate, the sporozoite rate and the EIR are too low to be indicators of the efficacy of control campaigns in this area.

How should nets be dried after insecticide treatment in the field?

Different procedures for drying polyamide (nylon) hammock nets (approximately 10 m2) posttreatment were evaluated. Forty-three nets were soaked for 10 min in a plastic bowl containing a solution of water and lambdacyhalothrin 0.05% (target dose of 10 mg/m2). Twenty-two freshly treated nets were put to dry in the sun: 13 of them hung vertically with the roof of the hammock net at the top and the hem at the bottom, and the other 9 were folded and laid flat on a plastic sheet. The rest of the nets (21) were dried in the shade: 11 of them hung vertically and 10 were folded flat on a plastic sheet. After 6-7 h, 2 15-cm2 samples of each net were cut, one on top of the lateral side and one 40 cm from the edge. Concentrations of lambdacyhalothrin were determined by gas chromatography-mass spectrometry on subsamples of netting (1 cm2). The mean insecticide concentrations of samples were compared between nets that were dried in the sun or in the shade, and placed to dry either hanging vertically or folded flat. In general, no differences were found between the mean insecticide concentrations of nets that were dried vertically in the sun or in the shade, or those dried flat in the shade. However, the mean dose was considerably lower in the remaining nets, which were dried flat in the sun. The reason for this is unclear but is hypothesized to be an effect of temperature. Results indicated that nets hung vertically either in the sun or shade dried faster than those laid flat. Based on the time to dry and mean insecticide extracted, it is recommended to hang treated nets vertically in the sun for drying.

Genetic relationships among Aedes aegypti collections in Venezuela as determined by mitochondrial DNA variation and nuclear single nucleotide polymorphisms.

A population genetic analysis of gene flow was conducted among 619 Aedes aegypti from nine collections distributed among six geographic regions of Venezuela. Genetic markers included a 387-basepair region of the mitochondrial NADH dehydrogenase 4 (ND4) gene and single nucleotide polymorphisms (SNPs) at 11 nuclear loci. Genotypes at SNP loci were identified using melting curve analysis. Six different ND4 haplotypes were detected and patterns of variation suggested that collections were isolated by distance. The variance in SNP allele frequencies was much less than the variance in haplotype frequencies and a pattern of isolation by distance was not detected. Aedes aegypti from eight collections were orally challenged with dengue 2 virus. Disseminated infection rates ranged from 77% to 95%. The percentage of mosquitoes exhibiting a midgut infection barrier ranged from 2% to 15%, and those exhibiting a midgut escape barrier ranged from 2% to 18%. Venezuelan Ae. aegypti appear to be susceptible to dengue virus infection.

Mosquito vector control and biology in Latin America--an 18th symposium.

The 18th Annual Latin American symposium presented by the American Mosquito Control Association (AMCA) was held as part of the 74th Annual Meeting in Sparks, NV, in March 2008. The principal objective, as for the previous 17 symposia, was to promote participation in the AMCA by vector control specialists, public health workers, and academicians from Latin America. This publication includes summaries of 35 presentations that were given orally in Spanish or presented as posters by participants from 5 countries in Latin America and Puerto Rico. Topics addressed in the symposium included surveillance, chemical and biological control, insecticide resistance, and repellency of Aedes aegypti; distribution, behavior, transmission of West Nile virus, and control of Culex; bionomics, ecology, and chemical and biological control of Anopheles vectors of malaria; insecticide resistance; and studies of Triatoma and Rhodnius.

Insecticide resistance levels and associated mechanisms in three Aedes aegypti populations from Venezuela

BACKGROUND The massive use of insecticides in public health has exerted selective pressure resulting in the development of resistance in Aedes aegypti to different insecticides in Venezuela. Between 2010 and 2020, the only insecticides available for vector control were the organophosphates (Ops) fenitrothion and temephos which were focally applied. OBJECTIVES To determine the state of insecticide resistance and to identify the possible biochemical and molecular mechanisms involved in three populations of Ae. aegypti from Venezuela. METHODS CDC bottle bioassays were conducted on Ae. aegypti collected between October 2019 and February 2020 in two hyperendemic localities for dengue in Aragua State and in a malaria endemic area in Bolívar State. Insecticide resistance mechanisms were studied using biochemical assays and polymerase chain reaction (PCR) to detect kdr mutations. FINDINGS Bioassays showed contrasting results among populations; Las Brisas was resistant to malathion, permethrin and deltamethrin, Urbanización 19 de Abril was resistant to permethrin and Nacupay to malathion. All populations showed significantly higher activity of mixed function oxidases and glutathione-S-transferases (GSTs) in comparison with the susceptible strain. The kdr mutations V410L, F1534C, and V1016I were detected in all populations, with F1534C at higher frequencies. MAIN CONCLUSION Insecticide resistance persists in three Ae. aegypti populations from Venezuela even in the relative absence of insecticide application.

Respuesta a atrayentes químicos y actividad horaria de Anopheles Meigen spp. (Diptera: Culicidae) en un área malárica del estado Bolívar, Venezuela / Response to chemical attractants and host seeking behavior of Anopheles Meigen spp. (Diptera: Culicidae) in a malaria endemic area of Bolívar State, Venezuela

Se evaluó la eficiencia de la trampa Mosquito Magnet Liberty PlusTM (MMLP) cebada con los atrayentes químicos 1-octen-3-ol (octenol) y Lurex 3TM (L-ácido láctico) junto con dióxido de carbono (CO2) para recolectar anofelinos en la localidad Ye'kwana de Boca de Nichare, municipio Sucre, estado Bolívar, Venezuela. La trampa fue operada entre las 18:00 y 06:00 horas, durante 10 noches por mes, durante cuatro meses (Julio ­ Octubre, 2015), alternándose cada noche el atrayente. A fin de caracterizar la actividad nocturna de los anofelinos, cada cuatro horas se cambiaba la jaulita donde eran atrapados los mosquitos. Se capturaron cinco especies de Anopheles y cuatro de Culicinae, siendo las más abundantes, Anopheles (Nyssorhynchus) darlingi Root y Anopheles (Nyssorhynchus) oswaldoi (Peryassú) sensu lato. El ANOVA factorial no paramétrico de Transformación de Rangos Alineados para la variable atrayente y sus interacciones con especie e intervalo horario no fueron estadísticamente significativas (p > 0.05). Estas especies muestran comportamientos diferentes: An. darlingi tiene el pico de actividad nocturna entre las 22:00 y 02:00 horas, mientras que en An. oswaldoi s.l. se observa un incremento de la actividad nocturna progresivamente que se mantiene durante el intervalo 02:00-06:00 horas. Los resultados permiten concluir que la trampa MMLP cebada con octenol o Lurex es un método alternativo de recolección de mosquitos para la vigilancia entomológica en áreas maláricas remotas con población indígena. La actividad horaria de los vectores An. darlingi y An. oswaldoi s.l. sugiere que el uso de mosquiteros tratados con insecticidas puede resultar un método efectivo para el control de la transmisión de malaria en esta zona del país(AU)

The efficiency of the Mosquito Magnet Liberty PlusTM (MMLP) trap was evaluated baited with the chemical attractants 1-octen-3-ol (octenol) and Lurex3M (L-lactic acid) together with carbon dioxide (CO2) to collect anophelines in the Amerindian village Boca de Nichare, Sucre municipality, Bolívar State, Venezuela. The trap was operated between 18:00 and 06: hours, 10 nights per month for four months (July-October 2015), the attractants were alternated nightly. To determine the host seeking behavior of anophelines, the trap cage was changed every 4 hours. Five species of Anopheles and four of Culicinae were collected; Anopheles (Nyssorhynchus) darlingi Root and Anopheles oswaldoi (Peryassú) sensu lato were the most abundant species collected. The Align Rank Transform (ART) test for nonparametric ANOVA for the variable attractant and its interactions with species and hour interval were not significant (p > 0.05). An. darlingi and An. oswaldoi s.l. showed contrasting host seeking activity: An. darlingi showed a peak between 22:00 and 02:00 hours, while in An. oswaldoi s.l. there was a steady increase in activity up to the interval 02:00-06:00 hours. The MMLP trap baited either with octenol or Lurex is an alternative method for entomological surveillance in remote Amerindian malaria endemic areas. The biting activity of the vectors An. darlingi and An. oswaldoi s.l. suggests that the use of long-lasting insecticide treated nets could be an effective method to control the transmission of malaria in this area(AU)

Detection of dengue viruses in field-caught Aedes aegypti (Diptera: Culicidae) in Maracay, Aragua state, Venezuela by type-specific polymerase chain reaction.

Virological surveillance of dengue viruses in Aedes aegypti populations constitutes a powerful tool for early prediction of dengue outbreaks. We have standardized a protocol for viral RNA extraction from individual and pools of mosquitoes that permits a sensitive detection of dengue virus without RNA degradation or PCR inhibition when we apply a semi-nested RT-PCR. The limit of detection for each dengue serotype was 0.1 PFU. In a prospective field study conducted from November 2000 to December 2001, adult female A. aegypti mosquitoes from several municipalities with high dengue transmission in Maracay, Aragua State, Venezuela were collected and screened for dengue viruses using RT-PCR. We analyzed a total of 296 A. aegypti pools (1,632 mosquitoes); of these, 154 pools (469 mosquitoes) were collected from houses with persons with clinical diagnosis of dengue (dengue houses), and 142 pools (1,163 mosquitoes) from adjacent residences (neighbour houses). From the dengue houses, eight mosquito pools (5.2%) were positive for DENV-1 (0.7%), DENV-3 (3.2%) and DENV-4 (1.3%) viruses. From the neighbour houses, 18 mosquito pools (12.7%) were positive for DENV-3 (12%) and DENV-4 (0.7%) viruses. From these 26 RT-PCR positive mosquito pools (containing 1-25 mosquitoes each), 22 pools (84.6%) were positive for DENV-3. The most prevalent serotype in the 2001 dengue outbreak was also DENV-3. The minimum infection rate in both A. aegypti collections, from dengue houses and neighbour houses was 17 and 15 per 1,000, respectively. The relevance of these results for dengue surveillance is discussed.

Malaria Control in Amerindian Communities of Venezuela : Strengthening Ecohealth Practice Throughout Conservation Science and Capability Approach.

Adaptive management and ecohealth frameworks were developed for malaria elimination in Amerindian riparian communities of Venezuela. These frameworks were developed as a strategy to capture, organize, and communicate connections among key factors related to local malaria complex systems. Important causal relationships between social, economic, and environmental stressors which are determinant of malaria were identified at different levels and assumptions that guide interventions are offered, based on available scientific knowledge and input from stakeholders. Drawing on our experience of action research committed to the health of Amerindian populations and conservation of areas with biodiversity value, the authors provide lessons to strengthen the practice of an ecohealth approach. First, conservation targets were considered as a way to achieve sustainable human well-being rather than as a consequence of well-being. Second, the effectiveness and sustainability of technical solutions generally proposed for malaria control depend largely on individual knowledge, attitudes, and practices. Hence, it is necessary to look at the real opportunities of choices that Amerindian people have for attaining a life without malaria, and therefore pay attention to local capabilities, needs, and freedom to choose. The ecohealth approach can benefit from the capability approach, and we explain why.

Morphological characters of adult Anopheles (Nyssorhynchus) marajoara in Venezuela.

A morphometric study was carried out to find diagnostic characters with which to update taxonomic keys for field identification of Anopheles (Nyssorhynchus) marajoara and the 3 other sympatric Anopheles (Nyssorhynchus) species (An. darlingi, An. argyritarsis, and An. braziliensis) that occur in Venezuela. Diagnostic random amplified polymorphic DNA-polymerase chain reaction markers from wild-caught specimens showed that An. marajoara was the only species in the Anopheles albitarsis complex collected in Venezuela.

A global map of dominant malaria vectors.

BACKGROUND: Global maps, in particular those based on vector distributions, have long been used to help visualise the global extent of malaria. Few, however, have been created with the support of a comprehensive and extensive evidence-based approach. METHODS: Here we describe the generation of a global map of the dominant vector species (DVS) of malaria that makes use of predicted distribution maps for individual species or species complexes. RESULTS: Our global map highlights the spatial variability in the complexity of the vector situation. In Africa, An. gambiae, An. arabiensis and An. funestus are co-dominant across much of the continent, whereas in the Asian-Pacific region there is a highly complex situation with multi-species coexistence and variable species dominance. CONCLUSIONS: The competence of the mapping methodology to accurately portray DVS distributions is discussed. The comprehensive and contemporary database of species-specific spatial occurrence (currently available on request) will be made directly available via the Malaria Atlas Project (MAP) website from early 2012.

Genetic structure of Latin American Aedes aegypti / Estructura genética de Aedes aegypti latinoamericano

The mosquito Aedes aegypti is the main Dengue vector in Latin America. This study investigated the genetic structure of this vector using samples collected in Venezuela, Colombia, Peru, Mexico, Argentina, Puerto Rico, and French Guiana. We examined the distribution of a 246-basepair region of the NADH dehydrogenase subunit 4 mitochondrial gene among a total of 369 Ae. aegypti from all the populations. This gene was amplified by the polymerase chain reaction and tested for variation using single strand conformation polymorphism analysis. Twelve haplotypes were detected among all the countries sampled and grouped into two clades. Significant differentiation was detected among the populations studied and these were not genetically isolated by distance.

El mosquito Aedes aegypti es el principal vector del Dengue en los países latinoamericanos. En este estudio se investigó la estructura genética de este vector en muestras colectadas en Venezuela, Colombia, Perú, Mexico, Argentina, Puerto Rico y Guyana Francesa. Nosotros examinamos la distribución de una región de 246 pares de bases del gen mitochondrial de la subunidad 4 de la NADH deshidrogenasa entre un total de 369 Ae. aegypti de todas las poblaciones. Este gen fue amplificado por la reacción en cadena de la polimerasa y la variación se determinó usando el análisis de polimorfismos de conformación de cadena simple. Doce haplotipos se detectaron entre todos los países y se repartieron en dos clados. Una diferenciación significativa se detectó entre las poblaciones y estas no se encontraron genéticamente aisladas por distancia.