Understanding the blood-feeding patterns of mosquitoes is essential for evaluating their potential as disease vectors, especially in urban areas where mosquitoes coexist with humans, domestic animals and wildlife. This study aimed to bridge a substantial gap in regional knowledge by identifying the blood meal sources of field-collected mosquitoes in domestic and open green environments from two urbanisations of temperate Argentina, the Área Metropolitana de Buenos Aires (AMBA) and Tandil, using molecular techniques. Female mosquitoes were collected from November 2019 to March 2020 and April–May 2021. A bipartite network analysis was performed for each environment and urbanisation. A total of 103 blood meals from Aedes (2 species) and Culex (7 species) were identified. Among these, five mammal and 18 bird species were recognised as hosts. Aedes mosquitoes exclusively fed on mammals, while Culex mosquitoes exhibited a broader host range including both birds and mammals. In AMBA, the open green environments were composed by more mosquito species than the domestic environments, while both presented similar numbers of vertebrate species. In contrast, in open green environments from Tandil only blood-fed Aedes albifasciatus were collected. For open green environments of AMBA and domestic environments of Tandil, results suggested some degree of host selection. For the three main vectors of diseases in the region, Aedes aegypti, Ae. albifasciatus and Culex pipiens molestus, we present the first molecular evidence of human blood meals in South America. Epidemiological significance of the present findings is discussed.

The vector role of Aedes aegypti for viral diseases including dengue and dengue hemorrhagic fever makes it imperative for its proper control. Despite various adopted control strategies, genetic control measures have been recently focused against this vector. CRISPR Cas9 system is a recent and most efficient gene editing tool to target the sex determination pathway genes in Ae. aegypti. In the present study, CRISPR Cas9 system was used to knockout Ae. aegypti doublesex (Aaedsx) and Ae. aegypti sexlethal (AaeSxl) genes in Ae. aegypti embryos. The injection mixes with Cas9 protein (333 ng ul−1) and gRNAs (each at 100 ng ul−1) were injected into eggs. Injected eggs were allowed to hatch at 26 ± 1°C, 60 ± 10% RH. The survival and mortality rate was recorded in knockout Aaedsx and AaeSxl. The results revealed that knockout produced low survival and high mortality. A significant percentage of eggs (38.33%) did not hatch as compared to control groups (P value 0.00). Highest larval mortality (11.66%) was found in the knockout of Aaedsx female isoform, whereas, the emergence of only male adults also showed that the knockout of Aaedsx (female isoform) does not produce male lethality. The survival (3.33%) of knockout for AaeSxl eggs to the normal adults suggested further study to investigate AaeSxl as an efficient upstream of Aaedsx to target for sex transformation in Ae. aegypti mosquitoes.

Dengue is a fast-spreading mosquito-borne viral disease in the world. The primary vector of the disease is Aedes aegypti of the family Culicidae. It is a container breeder. Since a vaccine or a drug has not been developed against dengue, vector control appears to be the best method so far to control dengue. The current study was conducted to determine the effect of delayed mating on fecundity, fertility, life-history parameters, and longevity of Ae. aegypti, because such information can help formulate integrated vector control strategies involving the release of sub-fertile males into the environment. During this study, mating was delayed by 0, 2, 5, and 8 days after emergence. Males and females were separated by hand at the pupal stage using the apparent size difference of the sexes. The separated pupae were kept in separate cages until emergence. When mating was delayed for 8 days, the number of eggs laid by the female declined by 38%, and the percentage number of eggs that hatched reduced by 24%. However, the percentage of larval mortality, duration of the larval and pupal periods, and adult longevity were not significantly affected. The current results indicate that delayed mating has a negative effect on the reproductive performance of vector mosquitoes.

Aedes aegypti (L.) (Diptera: Culicidae) is one of the main vectors of arboviruses, including dengue, Zika, and chikungunya. It almost exclusively inhabits urban areas. Both sexes feed on plant carbohydrates, although for males, this is their only food source. In the case of floral nectars, mosquitoes locate plant sugar sources assisted by volatile compounds. In this work, we found that the floral scent of Senecio madagascariensis elicited a behavioral response in males; therefore, we focused on identifying the volatiles emitted by these flowers. The terpenes (±)-α-pinene, β-pinene, sabinene, and phellandrene and 1-alkenes 1-undecene, and 1-nonene were identified. To determine which compounds are bioactive, pure synthetic lures were assessed using an olfactometer. Only the main compound 1-nonene was an attractant for males. Since our goal was the introduction of synthetic floral-based attractants in toxic sugar-baited traps, we formulated 1-nonene in solid paraffin and stearin matrices to obtain a controlled release system. The bioassay with a toxicological end point showed that the incorporation of a feeding attractant to the toxic sugar trap increased overall mortality. These results suggest that it is possible to use plant volatile compounds or flower cuttings as male Ae. aegypti attractants to improve the efficacy of baited traps.

A model for the Aedes aegypti lifecycle is developed that takes into account temperature-dependent maturation and death rates for several life stages, wet and dry egg oviposition with flooding, as well as three classes of larval habitat with different temperature profiles: outdoor (subject to external temperature fluctuations, human-inhabited), indoor (temperature moderated, human-inhabited, interior), and enclosed (temperature moderated, human free, exterior). An equilibrium analysis shows that the temperature range of outdoor viable equilibrium populations aligns closely with reported risk levels. Temperature patterns for El Paso, Texas; New York, New York; New Orleans, Louisiana; Orlando, Florida; and Miami, Florida, are considered. In four of these locations (all but New York), enclosed habitats can support mosquito populations even if all outdoor and indoor habitats are removed. In two locations (El Paso and New York) the model shows that in spite of the disappearance of adult mosquitoes during colder temperatures, populations reach seasonal steady state due to the survival of eggs. The results have implications for both vector and disease control.

Aedes-borne diseases are on the rampant rise despite continued application of chemical insecticide-based interventions. The appearance of high degree of insecticide resistance in Aedes species and noxious effects on environment and non-targets have raised further concerns. Among new chemical interventions, neonicotinoids are considered a safe and effective approach. The present study investigated the control potency of acetamiprid and development of resistance in Aedes aegypti larvae; and the involvement of CYP450 monooxygenases in inducing resistance. The early fourth instars of Ae. aegypti parent susceptible strain (PS) were selected with acetamiprid for 15 generations (ACSF strain) increasing the resistance to 19.74-fold in ACSF-10 and 36.71-fold in ACSF-15. The ACSF-10 larvae were assayed with acetamiprid combined with piperonyl butoxide (PBO) in three different ratios (1:1, 1:5 and 1:10) and selected for next five generations with 1:10 combination. Selection with synergized acetamiprid (APSF strains) reversed as well as reduced the rate of resistance development resulting in only 1.35-fold resistance in APSF-15. The APSF strains showed %monooxygenase dependency ranging from 86.71 to 96.72%. The estimation of the monooxygenases levels in parent and selected larvae showed increased monooxygenase level in the ACSF strains by 2.42–2.87-fold. The APSF-15 strains exhibited 57.95% lower enzyme production than ACSF-15 strain. The reduction and reversion of resistance by using PBO and the elevated levels of monooxygenases in ACSF and reduction in APSF strains recommend the involvement of CYP450-mediated mechanism in the development of acetamiprid resistance in Ae. aegypti. These studies could help in devising resistance management strategies in order to preserve the efficiency of pre-existing insecticides.

In Argentina, the mosquito Aedes aegypti (L.) (Diptera: Culicidae) is distributed from subtropical to temperate climates. Here, we hypothesized that the expansion of Ae. aegypti into colder regions is favoured by high-phenotypic plasticity and an adaptive inhibition of egg hatching at low temperatures. Thus, we investigated the hatching response of eggs of three populations: one from a subtropical region (Resistencia) and two from temperate regions (Buenos Aires City and San Bernardo) of Argentina. Eggs collected in the field were raised in three experimental colonies. F1 eggs were acclimated for 7 days prior to immersion at 7.6 or 22°C (control eggs). Five immersion temperatures were tested: 7.6, 10.3, 11.8, 14.1 and 16°C (range of mean winter temperatures of the three localities). A second immersion at 22°C was performed 2 weeks later to assess the inhibition to hatch under favourable conditions. After the first immersion, we compared the proportions of hatched eggs and dead larvae among treatment levels, whereas after the second immersion we compared the hatching response among the three populations. The factors that most influenced the egg hatching response were the geographical origin of the populations and the immersion temperature, but not the acclimation temperature. The proportions of hatching and larval mortality at low temperatures were higher for Resistencia than for Buenos Aires and San Bernardo, whereas the hatching response at ambient temperature was lower for San Bernardo than for Buenos Aires and Resistencia. The results support the hypothesis that populations from colder regions show an adaptive inhibition of egg hatching.

Aedes aegypti, historically known as yellow fever (YF) mosquito, transmits a great number of other viruses such as Dengue, West Nile, Chikungunya, Zika, Mayaro and perhaps Oropouche, among others. Well established in Africa and Asia, Aedes mosquitoes are now increasingly invading large parts of the American continent, and hence the risk of urban YF resurgence in the American cities should because of great concern to public health authorities. Although no new urban cycle of YF was reported in the Americas since the end of an Aedes eradication programme in the late 1950s, the high number of non-vaccinated individuals that visit endemic areas, that is, South American jungles where the sylvatic cycle of YF is transmitted by canopy mosquitoes, and return to Aedes-infested urban areas, increases the risk of resurgence of the urban cycle of YF. We present a method to estimate the risk of urban YF resurgence in dengue-endemic cities. This method consists in (1) to estimate the number of Aedes mosquitoes that explains a given dengue outbreak in a given region; (2) calculate the force of infection caused by the introduction of one infective individual per unit area in the endemic area under study; (3) using the above estimates, calculate the probability of at least one autochthonous YF case per unit area produced by one single viraemic traveller per unit area arriving from a YF endemic or epidemic sylvatic region at the city studied. We demonstrate that, provided the relative vector competence, here defined as the capacity to being infected and disseminate the virus, of Ae. aegypti is greater than 0.7 (with respect to dengue), one infected traveller can introduce urban YF in a dengue endemic area.

Since 2008, several outbreaks of yellow fever and dengue occurred in Abidjan, the economic capital of Côte d'Ivoire. A better knowledge of the biology of Aedes aegypti populations, the main vector of yellow fever and dengue viruses, is necessary to tailor vector control strategies implemented in the city. This study was designed to determine some biological parameters, occurring during the life cycle of two morphological phenotypes of Ae. aegypti larvae. Mosquitoes were sampled in a suburb of Abidjan (Treichville) using the WHO layer-traps technique. Biological parameters were studied in laboratory under standard conditions of temperature (27°C ± 2°C) and relative humidity (80% ± 10%). Our results indicated that the mean eggs laid by females from ‘brown larvae’ (BL) (85.95, 95% confidence interval (CI 95%) 78.87–93.02) was higher than those from ‘white larvae’ (WL) (64.40%, CI 95% 55.27–73.54). The gonotrophic cycle was 3 and 4 days in females from BL and WL, respectively. The overall yield of breeding mosquitoes from BL (63.88%, CI 95% 62.61–65.14) was higher compared with those of mosquitoes from WL (59.73%, CI 95% 58.35–61.12). The sex ratio (male/female) was 0.95 and 1.68 in Ae. aegypti populations from BL and WL, respectively. Females from BL lived slightly longer than those from WL (t = −2.332; P = 0.021). This study shows that Ae. Aegypti populations from BL and WL present different biological parameters during their life cycle. This could have an implication on their ability to transmit human disease viruses such as dengue and yellow fever. Further molecular studies are needed to determine genetic divergence between these Ae. aegypti populations.

The control of dengue constitutes a great challenge for public health; however, the methods normally used have shown themselves to be insufficient to keep the indices of infestation of Aedes aegypti under control. Recently, beyond the large number of cases and deaths associated with dengue, new risks have arisen such as those represented by chikungunya fever and Zika. In the light of the great significance of these problems within the public health context, two areas in a municipality in the interior of the State of São Paulo, Brazil were selected in 2014. One of them, Bairro Cidade Nova, was submitted to the conventional method of nebulization with portable equipment, and the other, Bairro Jardim Europa, received the application of the insecticide by means of heavy-equipment coupled to the vehicle. During the project, 1355 mosquito eggs were collected, 1105 of them in Bairro Cidade Nova and 205 in Bairro Jardim Europa. After the applications with heavy-equipment in the months of March and April, the number of cases of the disease reported in the month of April for Bairro Jardim Europa was less than half that of Bairro Cidade Nova, which had received the conventional treatment. The nebulization with the heavy-equipment may constitute a viable and effective strategy for achieving better results in the control of Ae. aegypti.

Buenos Aires city is located near the southern limit of the distribution of Aedes aegypti (Diptera: Culicidae). This study aimed to assess long-term variations in the abundance of Ae. aegypti in Buenos Aires in relation to changes in climatic conditions. Ae. aegypti weekly oviposition activity was analyzed and compared through nine warm seasons from 1998 to 2014, with 200 ovitraps placed across the whole extension of the city. The temporal and spatial dynamics of abundances were compared among seasons, and their relation with climatic variables were analyzed. Results showed a trend to higher peak abundances, a higher number of infested sites, and longer duration of the oviposition season through subsequent years, consistent with a long-term colonization process. In contrast, thermal favorability and rainfall pattern did not show a consistent trend of changes. The long-term increase in abundance, and the recently documented expansion of Ae. aegypti to colder areas of Buenos Aires province suggest that local populations might be adapting to lower temperature conditions. The steadily increasing abundances may have implications on the risk of dengue transmission.

The essential oil (EO) of Piper aduncum Linnaeus, known as ‘sireh lada’ to locals Malaysian, has the potential to be used as an alternative to synthetic insect repellents such as N,N-diethyl-meta-toluamide. However, the EO's efficacy as a repellent decreases after application due to the high volatility of its active ingredients. A number of studies have showed that optimizing the formulation of plant-based EOs can improve their efficacy as repellents. The present study sought to evaluate the effectiveness of 10% P. aduncum EO in ethanol and in three different semisolid formulations: ointment, cream and gel. These formulations were tested on Aedes aegypti under laboratory conditions. Each formulation was applied to the subject's hands, which were then inserted into a cage containing 25 nulliparous A. aegypti. The number of mosquitoes landing on or biting each subject's hand was recorded, and the repellency percentage, landing/biting percentage and protection time for each of the formulations were compared. There were no statistically significant differences between the semisolid EO formulations with regards to the repellency percentage and the landing/biting percentage at 4 h post-application. All three semisolid EO formulations were able to repel >65% of the A. aegypti at 4 h post-application. The EO ointment formulation provided a protection time (182.5 ± 16.01 min) that was statistically significantly longer than that associated with the EO gel formulation (97.5 ± 14.93 min). Meanwhile, the EO cream formulation provided a protection time of 162.5 ± 6.29 min. As the EO cream and ointment formulations displayed better repellent properties than the EO gel formulation, they appear to be the most promising P. aduncum EO formulations to be developed and commercialized as alternatives to synthetic repellents.

The Asian tiger mosquito, Aedes albopictus (Skuse) and the yellow fever mosquito, Aedes aegypti (Linnaeus) are medically important species that vector several arboviruses. Globally, populations of both species experience (and are sensitive to) photoperiodic variations. The present study aims to test if photoperiod regimes affect the population performance of Ae. albopictus and Ae. aegypti. Since mosquitoes have sex-specific strategies to maximize fitness, we also tested the hypothesis that Ae. albopictus and Ae. aegypti would exhibit differences in the male and female response (sexually dimorphic response) to various photoperiod treatments. We reared cohorts of first instar larvae to adulthood in three photoperiod treatments: short day (10 h light), control (12 h light) and long day (14 h light). We measured and compared survival to adulthood, population growth, development time of males and females, and wing length across treatments. Although we detected no effects of photoperiod on the population performance of both species, we found evidence of a sexual dimorphic response to photoperiod in Ae. albopictus, but not in Ae. aegypti, with Ae. albopictus females being more sensitive to variations in photoperiod. The observed differences between sexes of Ae. albopictus are consistent with sex-specific developmental constraints. The absence of a sexually dimorphic response to photoperiod in Ae. aegypti can be attributed to different strategies evolved in this species to prepare for unfavourable conditions associated with shorter day length. We discuss the ecological and medical implications of our findings.

A Bayesian Belief Network (BBN) for assessing the potential risk of dengue virus emergence and distribution in Western Australia (WA) is presented and used to identify possible hotspots of dengue outbreaks in summer and winter. The model assesses the probabilities of two kinds of events which must take place before an outbreak can occur: (1) introduction of the virus and mosquito vectors to places where human population densities are high; and (2) vector population growth rates as influenced by climatic factors. The results showed that if either Aedes aegypti or Ae. albopictus were to become established in WA, three centres in the northern part of the State (Kununurra, Fitzroy Crossing, Broome) would be at particular risk of experiencing an outbreak. The model can also be readily extended to predict the risk of introduction of other viruses carried by Aedes mosquitoes, such as yellow fever, chikungunya and Zika viruses.

Diversity amongst the different populations is one of the problems in the control of medically important mosquitoes. Wing morphometry was used to identify within-species variation amongst some populations of Aedes aegypti in three areas of Lagos State, Nigeria. Ten landmarks on the right wing of adult female Ae. aegypti mosquitoes were digitized using TPSdig software. Relative distances of the veins and interpoints were calculated as well as the Euclidean distances based on these relative distances. In three (50%) of the interpoints, the relative distances were significant (P<0.05) to both the Brown–Forsythe F ratio and Welch ANOVA. The highest Euclidean distance of 7.738 was between Ebute Metta 2 and Badagry; the lowest of 4.426 was between Ikorodu and Ebute Metta 2. The distance between Ebute Metta 1 and Ebute Metta 2 was 5.867, which is more than the distance between Ebute Metta 2 and Ikorodu. Three groups of Ae. aegypti were identified, and the differences in lengths and interpoints observed on the wing veins suggest a possible morphological divergence amongst populations.

Aquatic larval habitat conditions influence the development, fitness and vectorial capacity of mosquitoes. In turn, human activities can influence these conditions and shape mosquito vector distribution, thus affecting pathogen circulation and transmission. We measured environmental factors (chemical and microbial) in mosquito larval habitats and evaluated their potential as predictors for the occurrence of different mosquito species in an arbovirus and malaria endemic region of western Kenya. We found significantly greater proportions of Aedes aegypti mosquito larvae in ammonium and phosphate rich habitats, suggesting that fertilizer usage could potentially increase aquatic habitat suitability and Aedes vectored disease transmission. Anopheles gambiae larval habitats correlated significantly with higher temperatures. However, none of the Culex species’ habitats correlated with investigated variables, indicating greater larval plasticity compared to Aedes or Anopheles. Profiling of bacterial communities by 16S rRNA pyrosequencing revealed Proteobacteria, Bacterioidetes and Firmicutes as the major bacteria phyla present in mosquito aquatic microhabitats. Although there were no correlations between microbiota composition or diversity and larval species abundance, the dominant genera of microbes detected in larval habitats were reported in larval and adult mid-guts, suggesting that bacteria acquired from the larval habitats are transmitted to adult stages. This study identified the chemical and bacterial composition of aquatic microhabitats that are conducive to the development of different mosquito vectors in western Kenya. This information can inform potential vector control strategies by assessing management of breeding sites based on likely exposure to fertilizer, light and bacterial fauna.

Dengue has become the fastest-growing mosquito-borne disease in Sri Lanka and the control of the vectors, Aedes aegypti Linnaeus and Ae. albopictus Skuse, is the most effective way of controlling the disease. A detailed study on vector prevalence has not been recorded from Sri Lanka. Therefore, the present study was undertaken to study the prevalence of both vectors in four semi-urban study sites in two of the most affected districts, namely Kandy (wet zone) and Kurunegala (intermediate zone), by conducting egg surveys (using ovitraps) and larval surveys from June 2007 to May 2008. A total of 82,524 eggs and 2658 larvae of Ae. aegypti and Ae. albopictus were collected. A total of 3699 potential breeding habitats were examined. Ovitrap and larval indices (house, container and Breteau) showed that all four areas are at epidemic risk, especially due to a high abundance of Ae. albopictus. The highest numbers for both the species were from the Kandy sites where dense vegetation, high rainfall and low temperature prevailed. The results showed a high mortality rate during the egg-to-larva transition, suggesting that conducting an egg survey alone would overestimate the vector abundance and the disease risk. For Ae. albopictus, the monthly mean number of eggs showed positive relationships with relative humidity in both districts and with rainfall in the Kandy District. The number of dengue cases in the area had no positive relationships with the abundance of eggs or larval density indices. Discarded receptacles were the most preferred breeding habitat for these dengue vectors. Since the attractiveness (inferred by the calculated risk factors) of most of the breeding habitats was very high, elimination of these breeding sites is essential for the success of dengue control programmes.

Japan experienced dengue outbreaks vectored by Aedes albopictus during the Second World War. The probable vector density that caused the largest dengue outbreak in Nagasaki in 1942 was estimated using a mathematical simulation model. The estimated vector density was 15·0–558·0 per person when various assumptions of uncertain parameters were applied, such as proportion of symptomatic cases, vector mortality, and human biting rate of A. albopictus. When the most favourable disease spread conditions, such as a combination of the exclusive human biting rate and the longest vector survival were assumed, the vector density was 15–25 mosquitoes per person. Unusually high vector density due to wartime practices, and the traditional Japanese lifestyle were presumably responsible for the earlier dengue outbreak. If an outbreak occurs in present-day Japan, it is unlikely to spread as much as the previous one, as environmental conditions and human behaviour have changed in a protective manner.

Infecting Aedes aegypti mosquitoes with the bacteria Wolbachia has been proposed as an innovative new strategy to reduce the transmission of dengue fever. Field trials are currently being undertaken in Queensland, Australia. However, few mathematical models have been developed to consider the persistence of Wolbachia-infected mosquitoes in the wild. This paper develops a mathematical model to determine the persistence of Wolbachia-infected mosquitoes by considering the competition between Wolbachia-infected and non-Wolbachia mosquitoes. The model has four steady states that are biologically feasible: all mosquitoes dying out, only non-Wolbachia mosquitoes surviving, and two steady states where non-Wolbachia and Wolbachia-infected mosquitoes coexist. The stability of the steady states is determined with respect to the key parameters in the mosquito life cycle. A global sensitivity analysis of the model is also conducted. The results show that the persistence of Wolbachia-infected mosquitoes is dominated by the reproductive rate, death rate, maturation rate and maternal transmission. For the parameter values where Wolbachia persists, it dominates the population, and hence the introduction of Wolbachia has great potential to reduce dengue transmission.

The circulatory system in insects is formed by a dorsal vessel (heart) that is a tubular structure that pumps haemolymph in the insect body cavity. Up until now, much is known about the structural organization of the heart in certain Brachycera (Diptera) (e.g. Drosophila). In contrast, little information is available regarding members of the Nematocera suborder (e.g. mosquitoes), in spite of their relevance to human health. Considering their importance, details of the heart morphology of six mosquito species (Aedes albopictus, Ae. aegypti, Ae. fluviatilis, Culex quinquefasciatus, Anopheles darlingi and An. aquasalis) were investigated by scanning electron microscopy (SEM). The heart of the adult mosquitoes, both males and females, consists of a dorsal tubular structure in association with pericardial cells located alongside the heart and alary muscles displaying ramifications that extend from the body wall to the heart surface. The SEM analysis also revealed in detail the organization of the heart, including paired valves on the heart wall (ostia). This study provides a detailed analysis of the mosquito heart microanatomy and, in our view, represents a significant contribution towards the understanding of structural–functional relationships associated with the heart of mosquito.