Dengue Vector Research Articles

Retraction Note to: Synthesis and characterization of silver nanoparticles using Gmelina asiatica leaf extract against filariasis, dengue, and malaria vector mosquitoes.

Retraction Note to: Synthesis and characterization of silver nanoparticles using Gmelina asiatica leaf extract against filariasis, dengue, and malaria vector mosquitoes.

Development of a localized Web-based GIS Visualization Platform as a tool for Disseminating Habitat Abundance of Aedes Mosquito Vectors in the Philippines

Abstract. Disseminating up-to-date information on dengue mosquito habitats aided by modern GIS approach would yield benefits to people being informed on the localized developments of the vector and therefore reduce people’s risk of being affected by the disease. This paper discusses the development of this monitoring and sensing mechanism through a Web-based GIS mapping and visualization. It utilizes the environmental, WHO dengue indices and weather information for a case of a city in the Philippines to visualize hotspot areas for habitats of dengue vectors. The paper would aid the government in deploying key assets and supporting DOH’s 5S drive or Search, Self, Seek, Support and Sustain strategy to curtail the spread of the disease. Through this paper, the author intends to develop a localized web-based platform Aedes mosquito vector. Presented in this study provides the data workflow from field observations and the inputs from environmental and meteorological data which has the potential to help enhance the data dissemination as well as the IEC campaign of local health leaders in reducing the mortality and morbidity of people affected by Dengue mosquitoes. This visualization platform includes an easy to set-up system using the available Web-based GIS which can be updated depending on the availability of field data and meteorological observations.

Heat Shock Proteins expression in malaria and dengue vector.

The survival of mosquitoes under changing climatic conditions particularly temperature, is known to be supported by Heat Shock Proteins (HSPs). In view of climate change, it is imperative to know whether the mosquito vectors will be able to withstand the increased temperatures or perish. Therefore, the present study was undertaken on the expression of HSPs' gene in An. stephensi and Ae. aegypti by exposing them to temperatures ranging from 5 to 45°C for 15-180-minutes for once and continuously or with rest in between. We compared the temperature-tolerance of both the vectors in terms of expression of HSP83, HSP70, and HSP26 genes at varying degrees of temperature and duration. HSP70 and HSP26 were found distinctively expressed in both the vectors as compared to HSP83. With continuous exposure up to 180-minutes at 35°C and 40°C, HSP70 was found upregulated up to 35 and 47 folds in Ae. aegypti while in An. stephensi, the expression was only 1 fold. Between the genes, HSP70 was highly expressed at different temperatures followed by HSP26 and HSP83. The manifold up-regulation of HSP genes in Ae. aegypti than An. stephensi may be attributed to the robustness of Aedes vector in terms of temperature tolerance. This study has shown that Ae. aegypti and An. stephensi can withstand considerable temperature stress by expressing HSPs when exposed to variable temperature and duration. In view of changing climate, the study provides a clue that the vector of dengue and zika virus will be difficult to control.

Monitoring of pyrethroid resistance in Aedes aegypti: first report of double and triple kdr mutations in Buenos Aires Province

BackgroundDengue is an emerging disease in Argentina due to the colonization of Aedes aegypti, the mosquito vector. Buenos Aires Province is the biggest and most populated district in Argentina, suffering dengue outbreaks of growing magnitude. During epidemic periods, pyrethroid insecticides are used in this country to control adult mosquitoes. Pyrethroid resistance in dengue vectors has been reported worldwide, making it necessary to implement resistance management strategies. The voltage-gated sodium channel is the target site of pyrethroids. Mutations in the gene encoding this protein, called kdr mutations, are usually the molecular cause of pyrethroid resistance in insects. In Ae. aegypti from the Americas, three kdr substitutions were described: V410L, V1016I, and F1534C. The diagnostic of kdr mutations is recommended for the early detection of pyrethroid resistance as well as the consequent planning of evidence-based control policies.MethodsWe distributed ovitraps across 16 localities in Buenos Aires Province, collecting 22,123 eggs. A total of 522 mosquitoes were genotyped in positions 1016 and 1534 of voltage-gated channel using multiplex high-resolution melting and/or TaqMan probe methods. A subset of 449 samples was also genotyped by a singleplex high-resolution melting method developed ad hoc and/or Sanger sequencing.ResultsWe have documented, for the first time to our knowledge in the central region of Argentina, the presence of the 1016Ikdr + 1534Ckdr allele. Additionally, our study reports the first identification of the V410L mutation in central Argentina. These results underscore a growing trend of pyrethroid resistance in Ae. aegypti, fueled by the widespread use of these insecticides.ConclusionsWe detected 1016Ikdr + 1534Ckdr and 410Lkdr mutations in central Argentina for the first time and improved the processivity and accuracy of kdr genotyping methods. The results are both a tool for resistance monitoring and a sign of alarm to direct efforts towards finding sustainable methods for vector control to complement or replace pyrethroids. Joint efforts between academia and authorities to develop and implement public policies for vector control are a productive way to transfer scientific results for their application in public health.Graphical

Spatial and temporal predictions of mosquito potential breeding sites and densities: integration of satellite imagery, in-situ data, and process-based modeling

Abstract. Dengue fever, primarily transmitted worldwide by the mosquito Aedes aegypti, poses significant public health challenges in tropical and subtropical regions. While effective vector control is crucial in the absence of reliable dengue vaccines, traditional control methods face obstacles like mosquito resistance to insecticides and a very high cost. By combining geospatial data, including satellite imagery, as descriptors, and entomological surveys as target variables in a Random Forest model, we predicted the number of potential mosquito breeding sites, derived the associated environmental carrying capacity for larvae, and used the Arbocarto process-based model to predict Ae.aegypti population densities in an urban region of French Guiana, South America. Our findings highlight that remote sensing data may help predict the number of potential breeding sites over urban areas. Our simulations indicate higher mosquito densities in urban residential areas and a strong spatial and temporal heterogeneity. These densities fluctuate according to intra-annual variations in temperature and precipitation, with higher densities associated with intermediate housing. A comparison with the conventional estimation of environmental carrying capacity for larvae in the current Arbocarto procedure highlights the advantages of our approach. Our study demonstrates the utility of integrating remote sensing with predictive modeling to enhance vector surveillance and control strategies, and provides a replicable approach for monitoring a dengue vector mosquito population in dynamic urban landscapes.

Impact of Nanoparticles Synthesized From Azadirachta indica (A. Juss) for Targeted Control of Malarial, Dengue, and Filariasis Vectors With Minimized Aquatic Toxicity

ABSTRACTIn the present study, biogenic silver nanoparticles (AgNPs) synthesized using Azadirachta indica leaf extract were evaluated for their toxic effects on both target and non‐target species. The botanical synthesized nanoparticles were tested against Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus and non‐targeted species Artemia salina and Eudrilus eugeniae at 24 h posttreatment. AgNPs were characterized using the following analytical techniques: UV‐Vis spectroscopy, FT‐IR, x‐ray diffraction (XRD), energy‐dispersive x‐ray spectroscopy (EDX), and scanning electron microscope (SEM) analysis. Results showed that AgNPs caused larvicidal activity with 90.66% mortality in An. stephensi, 97.33% in Ae. aegypti, and 93.33% in Cx. quinquefasciatus at 24 h posttreatment. A. indica‐derived AgNPs had lower LC50 (lethal concentration 50) and LC90 (lethal concentration 90) values of 44.803 and 252.886 ppm/mL in An. stephensi, 18.358 and 189.553 ppm/mL in Ae. Aegypti, and 36.492 and 219.800 ppm/mL in Cx. quinquefasciatus, respectively. Furthermore, AgNPs demonstrated lower toxicity effects with 50.66% mortality in A. salina and 38.66% mortality in E. eugeniae at 24 h posttreatment. Additionally, they exhibited lower LC50 and LC90 values of 240.996 and 533.618 ppm/mL in A. salina and 301.122 and 548.944 ppm/kg in E. eugeniae at 24 h posttreatment, respectively. The findings conclude that green‐synthesized AgNPs from plants offer a promising, cost‐effective, and target‐specific alternative for eco‐friendly mosquito larvicides. Future work should focus on developing these plant‐based agents with minimal non‐target toxicity to support sustainable pest control. Further research may explore large‐scale applications and assess the long‐term environmental impacts of AgNPs in integrated vector management programs.

Investigating the Determinants of Dengue Outbreak in Oman: A Study in Seeb.

This study investigates the key factors contributing to the dengue outbreak in Oman. Data on climate (e.g., temperature, humidity, wind pace), population traits (e.g., populace density), and vector dynamics (e.g., mosquito density) within the Seeb district of Oman from 2022 to 2023 were gathered. The partial least squares structural equation modeling (PLS-SEM) was performed to study which variables affect dengue outbreaks. The results indicate that climatic factors significantly affect the dengue vector (β = -0.361, p < 0.001) but do not directly impact the dengue outbreak. Population characteristics, however, have a more substantial impact on dengue transmission, with a total effect (β = 0.231, p = 0.002) being relatively higher than that of the vector itself (total effect: β = 0.116, p < 0.001). Even with ongoing vector intervention efforts, the study underscores the need to include innovative public health interventions when considering environmental and demographic factors. More advantageous surveillance and focused interventions in excessive-threat regions are essential to mitigate the effect of dengue in Oman.

Retraction Note to: Adulticidal properties of synthesized silver nanoparticles using leaf extracts of Feronia elephantum (Rutaceae) against filariasis, malaria, and dengue vector mosquitoes.

Retraction Note to: Adulticidal properties of synthesized silver nanoparticles using leaf extracts of Feronia elephantum (Rutaceae) against filariasis, malaria, and dengue vector mosquitoes.

Chemotactic activity of products of Elizabethkingia anophelis derived from Aedes albopictus against RAW264 murine macrophage cell line.

Dengue viruses enter dermal macrophages which are derived from other tissues after mosquito bites. We examined chemotactic factors derived from the dengue vector mosquito, Aedes albopictus, toward a RAW264 murine macrophage cell line. We found that Elizabethkingia anophelis that was isolated from mosquitoes exhibited migration-inducing activity toward RAW264 cells. The active substances were extracted using ethyl acetate to induce chemotactic movements. Chemotactic activity was eluted in the several fractions using the reversed-phase chromatography, suggesting that multiple substances were responsible for the activity. We isolated three bacterial colonies from the wild A. albopictus mosquitoes collected in Toyama Park, Tokyo prefecture, Japan. The bacterial 16S rRNA gene sequences were the most similar to those of Lonsdalea quercina. These bacteria also exhibited migration-inducing activity toward RAW264 cells. The migration-inducing activity of mosquito bacteria might be a new aspect of mosquito-mediated viral infections.

Urgent Need for Real Time Web Mapping GIS Applications to Dengue Surveillance for Epidemic Control and Management in India

Dengue epidemics has been increased both horizontal and vertical structure across the country since 2006, and hence, urgent need for GIS based dengue surveillance for epidemic control and management in the nation. The dengue epidemic cases and the mortality caused by dengue has become essentially public health important in India, and the situation has become worse since 2006. Dengue cases were clinically confirmed with thousands of cases and hundreds of deaths annually. The report of dengue epidemics is becoming daily news nowadays and it was reported from 24 states and Union Territories in India. The spatial and seasonal occurrence of dengue epidemics were reported across the country during the period of April to November annually, however, the vulnerability of epidemics was reported between the periods of mid-July to mid-November of every year. The geo-environmental and climate variables are fuelling to create a conducive environment for profusion of dengue vector mosquitoes Aedes species (Ae. aegypti or Ae. Albopictus). The information relevant to the geographical site specification of dengue vectors breeding habitats, vector abundance, vector density, etc., could be collected using global positioning systems (GPS), and these information are mapped and overlay on the thematic layers of climate variables (Temperature, relative humidity, saturation deficiency and Rainfall) under the geographical information systems (GIS) software platform for spatial analysis (cluster analysis, nearest neighborhood analysis, fussy analysis, probability of maximum and minimum likelihood analysis etc.,) for spatial risk prediction. The Google based internet GIS mapping could be used to embedded web mapping real time situation and updating the information with online data base connectivity (ODBC) within the GIS spatial engine for providing the real time information relevant to the dengue epidemics, and hence, the appropriate precaution measures could be taken to control dengue epidemics in the nation early in advance.

Eco-friendly synthesis of silver nanoparticles using Phyllanthus niruri leaf extract: Assessment of antimicrobial activity, effectiveness on tropical neglected mosquito vector control, and biocompatibility using a fibroblast cell line model

Abstract Mosquitoes are rapidly advancing as vectors of several severe diseases. The increasing resistance of mosquitoes and the environmental harm caused by insecticides pose significant challenges for eradicating mosquito vectors. In this study, 18 plant extracts were tested for larvicidal properties against Aedes aegypti, Culex quinquefasciatus, and Anopheles stephensi larvae. Phyllanthus niruri (Pn) showed enhanced larvicidal activity in both laboratory and field trials. The biosynthesis of silver nanoparticles (AgNPs) using Pn leaf methanol extract (Pn-LME) was confirmed by UV-visible spectroscopy, X-ray diffraction, transmission electron microscopy, scanning electron microscopy, and Fourier-transform infrared spectroscopy. Among various concentrations, 3 mM AgNPs exhibited significant LC90 values of 0.83, 1.46, and 9.11 ppm compared to 9.25, 93.48, and 14.60 ppm of Pn-LME against A. stephensi, C. quinquefasciatus, and A. aegypti, respectively. This indicates the high mortality of mosquito vectors at low AgNP concentrations. Additionally, Pn-AgNPs showed enhanced antibacterial activity and no cytotoxicity in normal fibroblast cells (L929). Field trials demonstrated a 98.70% decrease in mosquito larval density at A. stephensi breeding sites, a 96.55% reduction at C. quinquefasciatus sites, and a 97.85% reduction at Ae. aegypti sites. This study presents an eco-friendly and cost-effective AgNP bio-pesticide synthesized from Pn leaves for controlling and preventing the transmission of filarial, dengue, and malaria vectors.

Environmental pollution by plasticizers and the relationship to vector dengue mosquito Aedes aegypti: Bisphenol A (BPA) affect the development and viral immune pathway response

Bisphenol A (BPA) is a widely used industrial chemical found in various products, leading to human exposure through dietary and non-dietary sources. It acts as an endocrine disruptor, affecting reproductive processes in vertebrates by binding to estrogen receptors. While its effects on vertebrates have been extensively studied, much less is known about its impact on invertebrates. This study investigates the effects of BPA on the development and immune response of Aedes aegypti mosquitoes, which are important vectors for arboviral diseases, such as dengue, an emergent viral disease worldwide. Artificial aquatic niches (AAN) were sampled, and BPA concentrations were quantified. Ae. aegypti larvae were exposed to varying BPA concentrations, and their development, fecundity, fertility, and immune response were assessed. Results show delayed development and decreased emergence of mosquitoes exposed to BPA. Females exposed to BPA exhibited reduced oviposition while hatching rates remained unaffected. Furthermore, BPA exposure altered the expression of immune response genes in adult mosquitoes, with differential effects observed between sexes. This suggests that BPA exposure during early developmental stages can modulate the antiviral immune response in adult mosquitoes, possibly through the 20-hydroxyecdysone (20E) signaling pathway. Overall, this study highlights the potential impact of BPA on the development and immune function of mosquito vectors, with implications for vector competence and disease transmission.

Utilization of Gomophia egyptiaca (Red Starfish) in the Biological Control of Aedes aegypti Larvae

Background: Aedes aegypti mosquito spreads many vital diseases to humans. Traditionally, chemical insecticides were used for control. Recently, interest has grown in using aquatic predators, such as the Red Starfish (Gomophia egyptiaca), to target mosquito larvae in water, offering a potential biological control method. Methods: This study investigates the larvicidal potential of G. egyptiaca against Ae. aegypti larvae, a key vector for dengue and other mosquito-borne diseases after 24 hours. Result: The results demonstrate a significant increase in mortality rates with prolonged exposure to G. egyptiaca. After 2 hours, the mortality rate was 6.66%, increasing to 20% at 4 hours, 60% at 12 hours and peaking at 80% after 24 hours. Statistical analysis using one-way ANOVA revealed a highly significant p-value of 3.93, confirming the effectiveness of G. egyptiaca over time. LT50 was determined to be 14.91 hours, with a confidence interval of 12.40 to 18.65 hours. These findings indicate that G. egyptiaca could serve as a powerful alternative to chemical insecticides, offering a sustainable approach to mosquito control. However, field trials and evaluations of non-target impacts are necessary to fully assess its potential in diverse ecological settings. This study supports the integration of G. egyptiaca into mosquito management strategies, particularly where chemical resistance and environmental concerns are prevalent.

Addressing the rise of autochthonous vector-borne diseases in a warming Europe

The perspective explores the emergence of autochthonous tropical diseases in Europe, driven by climate change and the associated increase in vector-borne diseases. Rising temperatures, along with changes in humidity and rainfall patterns, have altered the activity, distribution, and diversity of vectors such as mosquitoes and ticks. Species like Aedes albopictus and Aedes aegypti, which are primary vectors of dengue, have established self-sustaining populations across Europe. This spread has led to a surge in cases of dengue fever, West Nile virus, and tick-borne encephalitis in temperate regions. The complexity of predicting these outbreaks is compounded by factors like vector diapause, serological cross-reactivity, and land-use changes. The perspective calls for the implementation of enhanced surveillance, weather-linked predictive models, and robust vector control strategies to mitigate the public health risks posed by the spread of these diseases. As climate change accelerates, Europe faces increasing health threats previously confined to tropical regions, emphasizing the need for proactive public health measures to protect populations from this growing threat.

Entomological Surveillance of Dengue Vector Aedes Mosquito Larvae in City Sadar Paharganj (SP) Zone of MCD, Delhi

Entomological Surveillance of Dengue Vector Aedes Mosquito Larvae in City Sadar Paharganj (SP) Zone of MCD, Delhi

Impact of compound warm and wet events on dengue fever infection in South and Southeast Asian countries

BackgroundMultiple studies have reported the profound influence of various climate factors on dengue fever infection, while the effects of joint exposure to warm and wet environment, a condition favouring dengue vectors, on disease transmission were less evaluated. This study aims to investigate the impact of various compound temperature, rainfall, and relative humidity exposures on dengue fever infection in the South and Southeast Asia regions. MethodsWeekly dengue fever surveillance data from 2012 to 2020 were collected from 48 locations in four countries named Singapore (1 location), Sri Lanka (15 locations), Malaysia (9 locations), and Thailand (23 locations, with 11 locations having different study periods). The distributed lag non-linear models were built to assess the impacts of compound temperature, rainfall, and relative humidity exposures on dengue fever infection risks. ResultsA total of 1,359,993 dengue fever cases were reported with 9.33%, 24.02%, 48.73%, and 17.91% cases contributed by Singapore, Sri Lanka, Malaysia, and Thailand, respectively. Compared to non-warm-non-wet, compound warm-wet was associated with an increased dengue risk (RR:1.32, 95% CI:1.21–1.44). Compared to moderate temperature-humidity, warm-wet environment was also associated with an increase in dengue risk (RR:1.37, 95% CI:1.22–1.55). In comparison to weeks with moderate temperature-rainfall, warm-wet weeks was linked to an elevated dengue risk (RR:1.39, 95% CI:1.27–1.52), whereas cold-dry weather would significantly reduce the infection risk (RR:0.70, 95% CI:0.62–0.80). Modification effects showed that the hot effect on dengue infection was more pronounced under higher humidity, while the impact of rainfall increased with warmer temperature. ConclusionWarm-wet events were associated with an increased dengue fever risk, while the infection risk would decline in cold-dry environment, and modification effects exist among exposures. Findings from this study highlight the importance of considering joint temperature, humidity, and rainfall dependency of dengue fever infection in disease prevention and control.

Bacillus thuringiensis israelensis VCRC B650 culture filtrate useful for mosquito oviposition attractant and larvicidal action

Oviposition attractants hold the potential attention for monitoring and controlling mosquitoes by enticing them to deposit eggs at specific locations. In this study, the bacterial culture filtrate of Bacillus thuringiensis var. israelensis VCRC B650, previously isolated from clay soil at an agricultural site in U.T of Puducherry, was investigated for the first time. This larvicidal bacterium was assessed for oviposition attractancy against dengue and filarial vectors, namely, gravid females of Aedes aegypti and Culex quinquefasciatus, respectively. The investigation revealed that, at 1:1 dilution of bacterial culture filtrate and water, Cx. quinquefasciatus exhibited significant attraction, with an oviposition activity index (OAI) value of +0.92, surpassing the standard threshold value of +0.30. The OAI of Bti culture filtrate at a 1:1 dilution was also compared with a standard oviposition attractant, p-cresol, at a concentration of 10ppm. It was observed that the Bti culture filtrate demonstrated a significant oviposition attractant effect, with 80 per cent more egg rafts laid, compared to p-cresol at 20 per cent. GC-MS analysis showed that there were 25 compounds present in the Bti culture supernatant, and out of those, only one compound, namely, benzaldehyde, was effective in showing oviposition attraction.

Wolbachia Natural Infection of Mosquitoes in French Guiana: Prevalence, Distribution, and Genotyping

Wolbachia are the most spread bacterial endosymbionts in the world. These bacteria can manipulate host reproduction or block virus transmission in mosquitoes. For this reason, Wolbachia-based strategies for vector control are seriously considered or have already been applied in several countries around the world. In South America, Wolbachia have been studied in human pathogen vectors such as sand flies and mosquitoes. In French Guiana, the diversity and distribution of Wolbachia are not well known in mosquitoes. In this study, we screened for Wolbachia natural infection in mosquitoes in French Guiana by using 16S rRNA, Wolbachia surface protein (WSP), and multi-locus sequence typing (MLST) molecular assays. A total of 29 out of 44 (65.9%) mosquito species were positive for natural Wolbachia infection according to the PCR results, and two Wolbachia strains co-infected three specimens of Mansonia titillans. Then, we analyzed the phylogenetic relationships among the Wolbachia detected. All of the tested specimens of Aedes aegypti, the major dengue vector of French Guiana, were negative. These results regarding Wolbachia strain, distribution, and prevalence in mosquitoes from French Guiana highlight Wolbachia–mosquito associations and pave the way for a future Wolbachia-based strategy for vector control in this Amazonian territory.

Context-specific variation in life history traits and behavior of Aedes aegypti mosquitoes.

Aedes aegypti, the vector for dengue, chikungunya, yellow fever, and Zika, poses a growing global epidemiological risk. Despite extensive research on Ae. aegypti's life history traits and behavior, critical knowledge gaps persist, particularly in integrating these findings across varied experimental contexts. The plasticity of Ae. aegypti's traits throughout its life cycle allows dynamic responses to environmental changes, yet understanding these variations within heterogeneous study designs remains challenging. A critical aspect often overlooked is the impact of using lab-adapted lines of Ae. aegypti, which may have evolved under laboratory conditions, potentially altering their life history traits and behavioral responses compared to wild populations. Therefore, incorporating field-derived populations in experimental designs is essential to capture the natural variability and adaptability of Ae. aegypti. The relationship between larval growing conditions and adult traits and behavior is significantly influenced by the specific context in which mosquitoes are studied. Laboratory conditions may not replicate the ecological complexities faced by wild populations, leading to discrepancies in observed traits and behavior. These discrepancies highlight the need for ecologically relevant experimental conditions, allowing mosquito traits and behavior to reflect field distributions. One effective approach is semi-field studies involving field-collected mosquitoes housed for fewer generations in the lab under ecologically relevant conditions. This growing trend provides researchers with the desired control over experimental conditions while maintaining the genetic diversity of field populations. By focusing on variations in life history traits and behavioral plasticity within these varied contexts, this review highlights the intricate relationship between larval growing conditions and adult traits and behavior. It underscores the significance of transstadial effects and the necessity of adopting study designs and reporting practices that acknowledge plasticity in adult traits and behavior, considering variations due to larval rearing conditions. Embracing such approaches paves the way for a comprehensive understanding of contextual variations in mosquito life history traits and behavior. This integrated perspective enables the synthesis of research findings across laboratory, semi-field, and field-based investigations, which is crucial for devising targeted intervention strategies tailored to specific ecological contexts to combat the health threat posed by this formidable disease vector effectively.

Synergistic Larvicidal and Pupicidal Toxicity and the Morphological Impact of the Dengue Vector (Aedes aegypti) Induced by Geranial and trans-Cinnamaldehyde.

Monoterpenes are effective and eco-friendly alternatives to conventional chemical larvicides. We tested single and binary mixtures of monoterpenes-geranial and trans-cinnamaldehyde-for their larvicidal and pupicidal activities against Aedes aegypti L. and for non-target toxicity on guppies (Poecilia reticulata Peters), using 1% (w/w) temephos as a reference. Geranial and trans-cinnamaldehyde at 250 ppm showed stronger larvicidal and pupicidal activities with a 100% mortality rate and an LT50 ranging from 0.3 to 0.6 h. All combinations were strongly synergistic against larvae and pupae compared to single formulations, with an increased mortality value (IMV) of 6% to 93%. The combination of geranial + trans-cinnamaldehyde (1:1) at 200 ppm showed the highest impact, with an IMV of 93%. The strongest larvicidal and pupicidal activities, a 100% mortality rate, and an LT50 of 0.2 h were achieved by geranial + trans-cinnamaldehyde (1:1) 500 ppm. They were thirty times more effective than a 1% temephos solution (LT50 ranging from 6.7 to 96 h) and caused obviously shriveled cuticles and a swollen respiratory system. All single and binary mixtures were not toxic to the guppies. Thus, the combination of geranial + trans-cinnamaldehyde has great potential as a safe insecticide for controlling mosquito larvae and pupae.