Larval Competition between Aedes and Culex Mosquitoes Carries over to Higher Arboviral Infection during Their Adult Stage.

The common house mosquito (Culex pipiens) is a native vector for West Nile virus (WNV). Invasive species like the tiger mosquito (Aedes albopictus) and Asian bush mosquito (Aedes japonicus) are rapidly spreading through Europe, posing a major threat as vectors for dengue, chikungunya (CHIKV), and Japanese encephalitis virus (JEV). These mosquitoes share a similar ecological niche as larvae, but the carry-over effects of aquatic larval interactions to the terrestrial adult stage remain largely unknown and their medical relevance requires further investigation. This study examines the context dependency of larval interactions among Aedes albopictus, Aedes japonicus, and Culex pipiens. The survival, development time, growth, and energetic storage were measured in different European populations within density-response (intraspecific) experiments and replacement (interspecific) experiments at 20 °C and 26 °C. Overall, Ae. japonicus was the weakest competitor, while competition between Ae. albopictus and Cx. pipiens varied with temperature. Adults emerging from this larval competition were infected as follows: Culex pipiens with WNV, Ae. albopictus with CHIKV, and Ae. japonicus with JEV. While no JEV infection was observed, mosquitoes experiencing interspecific interactions during their larval stages exhibited higher infection rates and viral RNA titers for CHIKV and WNV. This increased susceptibility to viral infection after larval competition suggests a higher risk of arbovirus transmission in co-occurring populations.

Chitin synthase genes of Aedes albopictus and their effects on development of pupae.

The invasive species Aedes albopictus is a major vector of several arboviruses. The global spread of this species seriously threatens human health. Insecticide resistance is an increasing problem worldwide that limits the efficacy of mosquito control. As the major structural component of cuticles, chitin is indispensable to insects. Chitin synthase (CHS) is the enzyme that catalyzes the biosynthesis of chitin at the final step. In this study, two CHS genes of Aedes albopictus (AaCHS1 and AaCHS2) were identified and their basic characteristics were evaluated via bioinformatics analysis. The highest abundance of AaCHS1 transcripts was detected in pupae, whereas that of AaCHS2 transcripts was detected in females; the highest expression levels of AaCHS1 and AaCHS2 were found in the epidermis and the midgut of pupae, respectively. The survival and emergence rates of pupae were significantly reduced after the injection of double-stranded RNA of AaCHS1 or AaCHS2, indicating that both AaCHS1 and AaCHS2 play crucial roles in the pupal development. In addition, the chitin content of pupae was obviously decreased after the suppression of AaCHS1 expression by RNA interference (RNAi) treatment. This influence of the RNAi treatment was further supported by the reduced chitin thickness and weakened chitin fluorescence signal in the new cuticle. The midgut of pupae presented a reduced intensity of the chitin fluorescence signal along with RNAi treatment specific to AaCHS2 expression. The results of this study indicate that CHS genes may be suitable as molecular targets used for controlling mosquitoes.

Hybridization between Aedes aegypti and Aedes mascarensis mosquitoes leads to disruption of male sex determination.

Understanding the sex determination pathway and its disruptions in mosquitoes is critical for the effective control of disease vectors through genetic manipulations based on sex separation. When male hybrids of Aedes aegypti females and Ae. mascarensis males are backcrossed to Ae. aegypti females, a portion of the backcross progeny manifests as males with abnormal sexual differentiation. We discovered a significant correlation between pupal abnormalities and the feminization of subsequent adults exemplified by the relative abundance of ovarian and testicular tissues. All intersex individuals were genetic males as they expressed a male determining factor, Nix. Further, our analysis of the sex-specific splicing of doublesex and fruitless transcripts demonstrated the presence of both male and female splice variants indicating that sex determination is disrupted. A comparative transcriptomic analysis revealed similar expression levels of most female-associated genes in reproductive organs and carcasses between intersexual males and normal females. Moreover, intersexes had largely normal gene expression in testes but significant gene downregulation in male accessory glands when compared with normal males. We conclude that evolving hybrid incompatibilities between Ae. aegypti and Ae. mascarensis involve disruption of sex determination and are accompanied by changes in gene expression associated with sexual differentiation.

Morphological changes in eggs and embryos of Aedes aegypti (Diptera: Culicidae) exposed to predicted climatic scenarios for the year 2100 in the Central Amazon.

According to the IPCC, by the year 2100, rises in global temperature could reach up to 5 °C above current averages. On a planet-wide scale, this is one of the effects of climate changes that could have repercussions on the biological cycle of Aedes aegypti, the main arbovirus vector in urban environments and a transmitter of the arboviruses that cause dengue, Zika, chikungunya and urban yellow fever. The objective of this study was to evaluate morphological changes in Ae. aegypti eggs and embryos maintained in a climate change simulator. For this, specimens obtained from an insectarium were kept in four chambers that simulated the range of environmental scenarios predicted by the IPCC for the year 2100. The eggs obtained from each room were collected and transported to the laboratory for morphometric and morphological analysis, using confocal and scanning microscopy. Aedes aegypti eggs (n=20) were used to obtain the following variables: total width, total length, length-width ratio and diameter of the micropylar disc. Additionally, 20 embryos were used to obtain the data on head capsule length, width and length-width ratio. The data were subjected to a normality test and the means of each variable were compared using ANOVA and Tukey's post-hoc test, considering (p ≤ 0.05). A significant reduction (p < 0.05) was observed mainly in the mean lengths under the current-extreme scenario (587.5 and 553.6 µm, respectively), as well as in the widths under the current-mild scenario (171 and 158.4 µm, respectively). The length of the cephalic capsule was also affected, showing significant differences in the means under the current-intermediate scenario (189.5 and 208.5 µm, respectively), as well as in the widths between the current-intermediate scenarios (173.7 and 194.9 µm, respectively). The results suggest significant changes in the morphometry of Ae. aegypti eggs and embryos as a result of the climatic influences to which the adults were subjected, which may have an impact on vector population density and, consequently, on arbovirus dynamics in urban environments.

The potential habitat and environmental fitness change of Aedes albopictus in Western Eurasia for 2081-2100.

BACKGROUND OBJECTIVES: The range of Aedes albopictus, the most important vector mosquito in Western Eurasia is growing due to climate change. However, it is not known how it will influence the habitats occupied by the species and its environmental fitness within its future range. METHODS: To study this question, the habitat characteristic of the mosquito was investigated for 2081-2100. RESULTS: The models suggest a notable future spread of the mosquito in the direction of Northern Europe and the parallel northward and westward shift of the southern and eastern potential occurrences of the mosquito. The models suggest a notable increase in generation numbers in the warmest quarter, which can reach 4-5 generations in the peri-Mediterranean region. However, both the joint survival rate of larvae and pupae and the number of survival days of adults in the warmest quarter exhibit decreasing values, as does the potential disappearance of the mosquito in the southern regions of Europe and Asia Minor, along with the growing atmospheric CO2 concentration-based scenarios. INTERPRETATION CONCLUSION: While in 1970-2000 Aedes albopictus mainly occupied the hot and warm summer temperate regions of Europe, the species will inhabit dominantly the cool summer temperate (oceanic) and the humid continental climate territories of North and North-Eastern Europe in 2081-2100.

Juvenile Hormone as a contributing factor in establishing midgut microbiota for fecundity and fitness enhancement in adult female Aedes aegypti.

Understanding the factors influencing mosquitoes' fecundity and longevity is important for designing better and more sustainable vector control strategies, as these parameters can impact their vectorial capacity. Here, we address how mating affects midgut growth in Aedes aegypti, what role Juvenile Hormone (JH) plays in this process, and how it impacts the mosquito's immune response and microbiota. Our findings reveal that mating and JH induce midgut growth. Additionally, the establishment of a native bacterial population in the midgut due to JH-dependent suppression of the immune response has important reproductive outcomes. Specific downregulation of AMPs with an increase in bacteria abundance in the gut results in increased egg counts and longer lifespans. Overall, these findings provide evidence of a cross-talk between JH response, gut epithelial tissue, cell cycle regulation, and the mechanisms governing the trade-offs between nutrition, immunity, and reproduction at the cellular level in the mosquito gut.

The effects of nano- and microplastic ingestion on the survivorship and reproduction of Aedes aegypti and Aedes albopictus (Diptera: Culicidae).

Microplastics (MPs) and nanoplastics (NPs) are pervasive environmental pollutants that are commonly ingested by organisms at different trophic levels. While the effects of MPs on aquatic organisms have been extensively studied, the impacts of MP ingestion on the host fitness of terrestrial organisms, mainly insects, have been relatively unexplored. This study investigates the effects of MP and NP ingestion on the survivorship and reproduction of 2 medically important mosquito species, Aedes aegypti Linnaeus (Diptera: Culicidae) and Aedes albopictus Skuse (Diptera: Culicidae). Larval and pupal survivorship of Ae. albopictus were not significantly affected by particle size or concentration, but there was a reduction of Ae. aegypti pupal survivorship associated with the ingestion of 0.03 µm NPs. In addition, there was little observed impact of 0.03 µm NP and 1.0 µm MP ingestion on adult survivorship, fecundity, and longevity. To further investigate the effects of MP ingestion on mosquito fitness, we also examined the effects of MPs of varying shape, size, and plastic polymer type on Ae. aegypti immature and adult survivorship. The data suggest that the polymer type and shape did not impact Ae. aegypti immature or adult survivorship. These findings highlight that understanding the effects of microplastic ingestion by mosquitoes may be complicated by the size, composition, and amount ingested.

Habituation leads to short but not long term memory formation in mosquito larvae.

In animals, memory allows to remember important locations and conserve energy by not responding to irrelevant stimuli. However, memory formation and maintenance are metabolically costly, making it worthwhile to understand the mechanisms underlying different types of memory and their adaptive value. In this study, we investigated the memory persistence of Aedes aegypti mosquito larvae, after habituation to a visual stimulus. We used an automated tracking system for quantifying the response of mosquito larvae to the passage of a shadow, simulating an approaching predator. First, we compared different retention times, from 4 min to 24 h, and found that mosquito larvae only exhibited memory capabilities less than 3 h after training. Secondly, we investigated the role of inter-trial intervals in memory formation. In contrast to other aquatic invertebrates, mosquito larvae showed no long-term memory even at long inter-trial intervals (i.e., 5 min and 10 min). Our results are discussed in relation to the ecological constraints.

Exposure to low-concentration fipronil impairs survival, behavior, midgut morphology and physiology of Aedes aegypti larvae.

The Aedes aegypti mosquito is a vector for various arboviruses, including dengue and yellow fever. Insecticides, such as pyrethroids and organophosphates, are widely used to manage and control these insects. However, mosquitoes have developed resistance to these chemicals. Therefore, this study aimed to investigate the effects of the commercial formulation of fipronil (Tuit® Florestal; 80% purity) on the survival, behavior, morphology, and proteins related to signaling pathways of the midgut in A. aegypti larvae under controlled laboratory conditions. Significant reductions in immature survival were observed in all concentrations of fipronil tested. Low insecticide concentration (0.5 ppb) led to decreased locomotor activity in the larvae and caused disorganization of the epithelial tissue in the midgut. Moreover, exposure to the insecticide decreased the activity of detoxifying enzymes such as catalase, superoxide dismutase, and glutathione-S-transferase. On the other hand, the insecticide increased protein oxidation and nitric oxide levels. The detection of LC3, caspase-3, and JNK proteins, related to autophagy and apoptosis, increased after exposure. However, there was a decrease in the positive cells for ERK 1/2. Furthermore, the treatment with fipronil decreased the number of positive cells for the proteins FMRF, Prospero, PH3, Wg, Armadillo, Notch, and Delta, which are related to cell proliferation and differentiation. These findings demonstrate that even at low concentrations, fipronil exerts larvicidal effects on A. aegypti by affecting behavior and enzymatic detoxification, inducing protein oxidation, free radical generation, midgut damage and cell death, and inhibiting cell proliferation and differentiation. Thus, this insecticide may represent a viable alternative for controlling the spread of this vector.

Design, Synthesis, and Bioactivity of Trifluoroethylthio-Substituted Phenylpyrazole Derivatives.

As the first marketed phenylpyrazole insecticide, fipronil exhibited remarkable broad-spectrum insecticidal activity. However, it poses a significant threat to aquatic organisms and bees due to its high toxicity. Herein, 35 phenylpyrazole derivatives containing a trifluoroethylthio group on the 4 position of the pyrazole ring were designed and synthesized. The predicted physicochemical properties of all of the compounds were within a reasonable range. The biological assay results revealed that compound 7 showed 69.7% lethality against Aedes albopictus (A. albopictus) at the concentration of 0.125 mg/L. Compounds 7, 7g, 8d, and 10j showed superior insecticidal activity for the control of Plutella xylostella (P. xylostella). Notably, compound 7 showed similar insecticidal activity against Aphis craccivora (A. craccivora) compared with fipronil. Potential surface calculation and molecular docking suggested that different lipophilicity and binding models to the Musca domestica (M. domestica) gamma-aminobutyric acid receptors may be responsible for the decreased activity of the tested derivatives. Toxicity tests indicated that compound 8d (LC50 = 14.28 mg/L) induced obviously 14-fold lower toxicity than fipronil (LC50 = 1.05 mg/L) on embryonic-juvenile zebrafish development.

Projection of dengue fever transmissibility under climate change in South and Southeast Asian countries.

Vector-borne infectious disease such as dengue fever (DF) has spread rapidly due to more suitable living environments. Considering the limited studies investigating the disease spread under climate change in South and Southeast Asia, this study aimed to project the DF transmission potential in 30 locations across four South and Southeast Asian countries. In this study, weekly DF incidence data, daily mean temperature, and rainfall data in 30 locations in Singapore, Sri Lanka, Malaysia, and Thailand from 2012 to 2020 were collected. The effects of temperature and rainfall on the time-varying reproduction number (Rt) of DF transmission were examined using generalized additive models. Projections of location-specific Rt from 2030s to 2090s were determined using projected temperature and rainfall under three Shared Socioeconomic Pathways (SSP126, SSP245, and SSP585), and the peak DF transmissibility and epidemic duration in the future were estimated. According to the results, the projected changes in the peak Rt and epidemic duration varied across locations, and the most significant change was observed under middle-to-high greenhouse gas emission scenarios. Under SSP585, the country-specific peak Rt was projected to decrease from 1.63 (95% confidence interval: 1.39-1.91), 2.60 (1.89-3.57), and 1.41 (1.22-1.64) in 2030s to 1.22 (0.98-1.51), 2.09 (1.26-3.47), and 1.37 (0.83-2.27) in 2090s in Singapore, Thailand, and Malaysia, respectively. Yet, the peak Rt in Sri Lanka changed slightly from 2030s to 2090s under SSP585. The epidemic duration in Singapore and Malaysia was projected to decline under SSP585. In conclusion, the change of peak DF transmission potential and disease outbreak duration would vary across locations, particularly under middle-to-high greenhouse gas emission scenarios. Interventions should be considered to slow down global warming as well as the potential increase in DF transmissibility in some locations of South and Southeast Asia.

Influence of insular conditions on wing phenotypic variation in two dominant mosquito vectors, Aedes albopictus and Armigeres subalbatus (Diptera: Culicidae), in the border archipelagos of Thailand.

Insects geographically separated into island and mainland populations often exhibit phenotypic variations, a phenomenon known as insular conditions. These conditions can lead to rapid evolutionary changes that affect the morphological characteristics of mosquito vectors. Nevertheless, studies that specifically examine phenotype differences between island and mainland mosquito populations have been limited. In this study, wing variation in size and shape was investigated using the geometric morphometric (GM) technique in two dominant mosquito vectors, Aedes albopictus and Armigeres subalbatus, in the Ranong and Trat archipelagos of Thailand. Significant differences in average wing centroid size (CS) were found in 6 out of 15 population pairs for Ae. albopictus (p < 0.05) and in 5 population pairs for Ar. subalbatus (p < 0.05). After removing the allometric effect, canonical variate analyses (CVA) based on wing shape analysis revealed overlap across all populations for both Ae. albopictus and Ar. subalbatus. However, the statistical analysis indicated that Ar. subalbatus exhibited wing shape differences across all populations (p < 0.05), and most Ae. albopictus populations also displayed distinct wing shapes (p < 0.05), except for the populations from Chang Island and the mainland of Ranong, which showed no significant differences (p > 0.05). These findings enhance our understanding of mosquito adaptability in island regions and provide valuable data for the surveillance and monitoring of vector evolution.

Ecotoxicological strategies employing biochemical markers and organisms to monitor the efficacy of malathion photolysis treatment.

The objective of this study was to assess the photolysis-mediated degradation of malathion in standard and commercial formulations, and to determine the toxicity of these degraded formulations. Degradation tests were carried out with 500 µg L-1 of malathion and repeated three times. The initial and residual toxicity was assessed by using Lactuca sativa seeds for phytotoxicity, Stegomyia aegypti larvae for acute toxicity, and Stegomyia aegypti mosquitoes (cultivated from the larval stage until emergence as mosquitoes) to evaluate the biochemical markers of sublethal concentrations. For the standard formulations the photolytic process efficiently reduced the initial concentration of malathion to levels below the regulatory limits however, the formation of byproducts was revealed by chromatography, which allowed for a more complete proposal of photolytic-mediated malathion degradation route. The degraded formulations inhibited the growth of L. sativa seeds, while only the untreated formulations showed larvicidal activity and mortality. Both formulations slightly inhibited acetylcholinesterase activity in S. aegypti mosquitoes, while the standard formulation decreased and the commercial formulation increased glutathione S-transferase activity. However, there were no significant differences for superoxide dismutase, esterase-α, esterase-ß and lipid peroxidation. These findings indicate that in the absence of the target compound, the presence of byproducts can alter the enzymatic activity. In general, photolysis effectively degrade malathion lower than the legislation values; however, longer treatment times must be evaluated for the commercial formulation.

Transmission dynamics of a reaction-advection-diffusion dengue fever model with seasonal developmental durations and intrinsic incubation periods.

In this paper, we propose a reaction-advection-diffusion dengue fever model with seasonal developmental durations and intrinsic incubation periods. Firstly, we establish the well-posedness of the model. Secondly, we define the basic reproduction number ℜ 0 for this model and show that ℜ 0 is a threshold parameter: if ℜ 0 < 1 , then the disease-free periodic solution is globally attractive; if ℜ 0 > 1 , the system is uniformly persistent. Thirdly, we study the global attractivity of the positive steady state when the spatial environment is homogeneous and the advection of mosquitoes is ignored. As an example, we use the model to investigate the dengue fever transmission case in Guangdong Province, China, and explore the impact of model parameters on ℜ 0 . Our findings indicate that ignoring seasonality may underestimate ℜ 0 . Additionally, the spatial heterogeneity of transmission may increase the risk of disease transmission, while the increase of seasonal developmental durations, intrinsic incubation periods and advection rates can all reduce the risk of disease transmission.

Oviposition site selection and subsequent offspring performance of Aedes aegypti in short- and long-term detritus accumulation conditions.

The "oviposition preference-offspring performance" hypothesis proposes that females lay their eggs in habitats that maximize the fitness of their offspring. The aim of this study was to assess the oviposition site selection by Aedes aegypti females and the success of their larvae in habitats with different detritus accumulation times, under conditions representative of the natural spatial variability of detritus quality and quantity in the Metropolitan Area of Buenos Aires, Argentina. Two experiments were performed, one assessing oviposition site selection and the other analyzing developmental success. In both experiments, two levels of detritus accumulation time were compared, one with short-time detritus accumulation (2 weeks), and the other with long-time detritus accumulation (8 weeks). Naturally fallen detritus was used in both experiments, collected in ten sites across the Metropolitan Area of Buenos Aires. In the oviposition experiment, two contiguous ovitraps corresponding to each accumulation time were placed at each of the ten sites and the number of eggs received for each accumulation time was compared. In the development experiment, always 19 larvae were raised in containers of both accumulation times and overall performance was compared using an integrated index that considers survival, development time and female size. A large variability in the amount of detritus collected at the different sites was observed. The number of eggs was significantly higher in the long-time than in the short-time detritus accumulation containers, and approximately proportional to the amount of detritus in each ovitrap. The performance was not affected by the detritus accumulation time, but a better performance was detected in containers that received a higher amount of organic detritus, regardless of the accumulation time. Leaves were on average the most abundant type of detritus, with an average of 53 % of the total detritus collected. The amount of leaves added 2 weeks before hatching showed a positive effect on larval performance. Our results do not support the "oviposition preference-offspring performance" hypothesis, since Ae. aegypti females laid eggs in containers where larvae did not show a better performance. Moreover, at larval densities related to the number of eggs actually laid in each of the accumulation times, it is expected that the performance would be even worse in the most selected containers, due to the density-dependent effects of crowding. Since the results obtained reflect the natural heterogeneity of the environmental conditions in the region studied, they might be a fairly good indicator of what occurs in natural larval habitats.

Temporal dynamics of the egg bank of Aedes aegypti (Diptera: Culicidae) in the winter-spring transition in a temperate region.

In temperate regions, the populations of Aedes aegypti (L.) (Diptera: Culicidae) remain in the egg stage during the winter. In Buenos Aires, Argentina, a temperate region in the southern limit of Ae. aegypti distribution, the start of the next reproductive season and the rate of increase of the adult population depend on the egg bank that remains after the winter. This study aimed to analyze the mortality, field-hatching, and survival of eggs exposed to field conditions representative of those that occur in nature. In addition, the post-exposure hatching response of the eggs was assessed. Four egg batches were exposed to natural conditions starting in mid-winter and were recovered progressively after 3, 6, 9, and 12 weeks. One egg batch (initial control) was not exposed in the field and remained under laboratory conditions. After the exposure period, the recovered intact, collapsed, and hatched eggs were counted. Intact eggs were immersed three times in the laboratory to study their hatching response. Progressive increases in the proportion of lost (presumably by predation), dead, and hatched eggs in successive egg batches were recorded. Field-hatching was recorded from late winter onwards. The first hatchings occurred in conditions probably not favorable to complete development into reproductive adults. A progressive decrease in live eggs was observed, with 51% of the recovered eggs alive after 12 weeks of exposure. In the laboratory, the hatching response in the first immersion was low for the initial control and for the eggs exposed for 3 weeks, and increased for successive cohorts. The results confirm that the survival of Ae. aegypti eggs in the winter-spring transition ensures persistence throughout the next favorable season in Buenos Aires City. The observed inhibition to hatch of the first batches might relate to a photoperiod-induced diapause, as observed in previous studies.

Using UAV images and deep learning in investigating potential breeding sites of Aedes albopictus.

Aedes albopictus (Diptera: Culicidae) plays a crucial role as a vector for mosquito-borne diseases like dengue and zika. Given the limited availability of effective vaccines, the prevention of Aedes-borne diseases mainly relies on extensive efforts in vector surveillance and control. In multiple mosquito control methods, the identification and elimination of potential breeding sites (PBS) for Aedes are recognized as effective methods for population control. Previous studies utilizing unmanned aerial vehicles (UAVs) and deep learning to identify PBS have primarily focused on large, regularly-shaped containers. However, there has been a small amount of empirical research into their practical application in the field. We have thus constructed a PBS dataset specifically tailored for Ae. albopictus, including items such as buckets, bowls, bins, aquatic plants, jars, lids, pots, boxes, and sinks that were common in the Yangtze River Basin in China. Then, a YOLO v7 model for identifying these PBS was developed. Finally, we recognized and labeled the area with the highest PBS density, as well as the subarea with the most urgent need for source reduction in the empirical region, by calculating the kernel density value. Based on the above research, we proposed a UAV-AI-based methodological framework to locate the spatial distribution of PBS, and conducted empirical research on Jinhulu New Village, a typical model community. The results revealed that the YOLO v7 model achieved an excellent result on the F1 score and mAP(both above 0.99), with 97% of PBS correctly located. The predicted distribution of different PBS categories in each subarea was completely consistent with true distribution; the five houses with the most PBS were correctly located. The results of the kernel density map indicate the subarea 4 with the highest density of PBS, where PBS needs to be removed or destroyed with immediate effect. These results demonstrate the reliability of the prediction results and the feasibility of the UAV-AI-based methodological framework. It can minimize repetitive labor, enhance efficiency, and provide guidance for the removal and destruction of PBS. The research can shed light on the investigation of mosquito PBS investigation both methodologically and practically.

Assessment of larval and pupal indices of dengue mosquito vectors in a North-Eastern state of Tripura, India.

BACKGROUND OBJECTIVES: Dengue is a major vector-borne disease having public health importance. It is caused by Dengue Virus (DENV) and is transmitted by mosquitoes of Aedes species. With the unavailability of a vaccine, vector control remains the only preventive measure for dengue. Studies have already been conducted to establish the presence of dengue vectors in the north-eastern states of India. However, limited studies have been conducted in Tripura state. In the present study we aimed to identify the preferred breeding habitats of dengue vectors in the state. METHODS: Clinical case data of dengue since the last five years was studied and the areas with the highest case numbers were identified. Entomological investigation was carried out in areas reporting the highest number of cases. Larvae were collected from the breeding habitats using standard protocol followed by morphological and molecular identification. Further, House index (HI), Container index (CI) and Pupal index (PI) were determined. The positive pools were then processed for incrimination for the presence of dengue virus. Calculation of entomological indices was done. RESULTS: Of the total 815 containers searched, 36.80% containers were positive for mosquito larvae. Among the immature mosquito collection, 836 adults emerged and were identified as Aedes albopictus using standard taxonomic keys followed by molecular methods. HI, CI and PI, varied from 15.38% to 100%, 21% to 31.04 %, and 2.93% to 110.53% respectively. However, none of the pools was positive for dengue virus. INTERPRETATION CONCLUSION: The present study identified Ae. albopictus as a potential vector of dengue in Tripura. The study gave important insights on the preferred larval habitats and provides information on the indication of displacement of Ae. albopictus from rural to urban and semi-urban areas. However, longitudinal studies for longer time frame are necessary for any conclusive remarks.

Evaluation of Insect Growth Regulators (IGRs) as biological pesticides for control of Aedes aegypti mosquitoes.

BACKGROUND OBJECTIVES: Insect growth regulators (IGRs) are biological hormone analogue or mimics used as pesticides to inhibit the growth of larva during their molting and skin shedding. This study aimed to test the effect of IGRs on the eggs hatching and post-hatching inhibition of Aedes mosquitoes and understanding its effect in the mosquito breeding habitats for reduction in adult emergence. METHODS: Experiments on the evaluation of three insect growth regulators (IGRs) for the control of different stages of Aedes aegypti was carried out during 2020-21. Each experiment consisted of four treatments viz., Pyriproxyfen, Novaluron, and Larvicol at 1.0 ppm and distilled water as a control. All experiments were carried out in completely randomized design (CRD) except eggs which were carried out in factorial design each with three replications. RESULTS: All tested IGRs performed better in affecting eggs, larval and pupal stages of Ae. aegypti. Highest eggs hatching inhibition (80%) of fresh eggs occurred in Pyriproxyfen followed by Novaluron (66%) and lowest in Larvicol (62%). Eggs hatch inhibition of embryonated eggs was lower than fresh eggs. Pyriproxyfen caused 69%, Novaluron 59% and Larvicol 39% eggs hatch inhibition of embryonated eggs. Both Pyriproxyfen and Novaluron performed better in causing 98-100% larval mortality followed by Larvicol (39%). Larval development to pupal stage was completely prevented by both Pyriproxyfen and Novaluron. Although Larvicol resulted in lowest eggs hatch and larval inhibition but prevented pupae to emerge as adults. Results further showed 70-89% mortality of 3rd instar larvae of Ae. aegypti when exposed to Pyriproxyfen and Novaluron solutions after 30 days storage at lab. temperature (27±2°C), RH 70±5. INTERPRETATION CONCLUSION: None of the IGRs was more effective at the pupal stage but showed carry-on activity of growth inhibition and mortality of the successive stages of development when used against eggs stages. Therefore, we recommend early application of IGRs at mosquito habitats during the beginning and onset of the season when very early stages of mosquitoes are available in the field.

Preparation of a long-lasting tablet of spinosad microspheres and its residual insecticidal efficacy against the yellow fever mosquito, Aedes aegypti (Diptera: Culicidae) larvae.

BACKGROUNDS: In order to provide a long-lasting formulation for spinosad (SP) targeting larval stages of Aedes aegypti (Linnaeus) and others alike, a SP tablet was developed based on microspheres, using polylactic acid as inside coating material. The microspheres were encapsulated using polyethylene glycol and 1-hexadecanol to form a sustained-release SP tablet. Micromorphology, active ingredient loading, structure identification, photolysis resistance and biological activity were evaluated in this report. RESULTS: (i) The SP microspheres had an average particle size of 6.16 ± 2.28 µm, low adhesion and good dispersion as evaluated by scanning electron microscopy and morphology. (ii) The average active ingredient loading and encapsulation of SP microspheres were 32.80 ± 0.74% and 78.41 ± 2.22%, respectively. (iii) The chemical structure of encapsulated SP was confirmed by Fourier transform infrared and 1H-nuclear magnetic resonance. (iv) The photostability of the microspheres and the tablets were evaluated. The results showed that DT50 (time required to dissipate 50% of the mass originally present) of SP was 0.95 days in microspheres and 6.94 days in tablets. (v) The long-term insecticidal activity of SP tablets was investigated, and the tablet had a long-lasting activity against the mosquito larvae, showing 100% larval mortality for 63 days. CONCLUSIONS: The study provided a new long-lasting formulation of SP, which displayed good efficacy in the control of Ae. aegypti larvae. © 2024 Society of Chemical Industry.