Hormone-dependent activation and repression of microRNAs by the ecdysone receptor in the dengue vector mosquito Aedes aegypti.

Female mosquitoes transmit numerous devastating human diseases because they require vertebrate blood meal for egg development. MicroRNAs (miRNAs) play critical roles across multiple reproductive processes in female Aedes aegypti mosquitoes. However, how miRNAs are controlled to coordinate their activity with the demands of mosquito reproduction remains largely unknown. We report that the ecdysone receptor (EcR)-mediated 20-hydroxyecdysone (20E) signaling regulates miRNA expression in female mosquitoes. EcR RNA-interference silencing linked to small RNA-sequencing analysis reveals that EcR not only activates but also represses miRNA expression in the female mosquito fat body, a functional analog of the vertebrate liver. EcR directly represses the expression of clustered miR-275 and miR-305 before blood feeding when the 20E titer is low, whereas it activates their expression in response to the increased 20E titer after a blood meal. Furthermore, we find that SMRTER, an insect analog of the vertebrate nuclear receptor corepressors SMRT and N-CoR, interacts with EcR in a 20E-sensitive manner and is required for EcR-mediated repression of miRNA expression in Ae. aegypti mosquitoes. In addition, we demonstrate that miR-275 and miR-305 directly target glutamate semialdehyde dehydrogenase and AAEL009899, respectively, to facilitate egg development. This study reveals a mechanism for how miRNAs are controlled by the 20E signaling pathway to coordinate their activity with the demands of mosquito reproduction.

A Challenge for a Unique Dengue Vector Control Programme: Assessment of the Spatial Variation of Insecticide Resistance Status amongst Aedes aegypti and Aedes albopictus Populations in Gampaha District, Sri Lanka.

BACKGROUND: To date, dengue is considered an important public health problem in Sri Lanka. Irrational use of insecticides without evidence-based applications has primed the development of resistance in mosquito vectors. METHOD: The present study investigated the resistance status of Aedes aegypti and Aedes albopictus to commonly used insecticides in three selected Medical Officer of Health (MOH) areas (i.e., Attanagalla, Dompe, and Negombo) in Gampaha District, Western Province of Sri Lanka. Entomological surveys were performed using ovitraps and larval collections. Larval bioassays were carried out to determine the LC50, LC90, and LC95 and susceptibility status for organophosphate temephos, whereas adult bioassays were performed to test the 0.03% deltamethrin and 0.8% malathion susceptibility. RESULTS: The study revealed that the temephos concentrations required to control Ae. aegypti (13.7-17.7 times) and Ae. albopictus (4.6-7.6 times) are higher than the diagnostic concentration (0.012 mg/L) proposed by the World Health Organization. The highest resistance levels were observed for both Ae. aegypti (14 ± 1.87) and Ae. albopictus (36 ± 1.87) collected from the Negombo MOH area. Therefore, the WHO recommended diagnostic concentration is no longer effective in controlling Ae. aegypti and Ae. albopictus larvae in these areas. Both the dengue vectors have evolved a high level of insecticide resistance to malathion and deltamethrin in the Gampaha District except Ae. albopictus mosquitoes in rural areas. Further, vectors in rural areas are indicated susceptible (>98%) to pyrethroids and emergence of resistance (<97%) for organophosphate insecticides. CONCLUSION: The results of this study warrant the vector management authorities on the proper application of insecticides and rational use in vector control. The susceptibility status of vector mosquitoes should be continuously monitored especially in dengue-endemic areas parallel to the routine surveillance programme. Further molecular studies are strongly recommended to determine the Knockdown Resistance (kdr) mutations among Aedes populations.

Case Report: Acute Vision Loss in a Young Returning Traveler with Dengue Fever.

Ocular complications are rare in patients with dengue fever, but may cause permanent loss of vision. We present the case of a 29-year-old German woman who developed severe acute vision loss because of dengue-associated maculopathy after traveling to Vietnam and Cambodia. Initially, the optical coherence tomography showed detachment of the retinal pigment epithelium, a central shift in the retinal pigmentation and intraretinal cysts. The patient was hospitalized and treated with a short course of intravenous prednisolone. Vision improved, and the patient showed full recovery at 9 months after the onset. This case highlights the importance of awareness and adequate management for ocular involvement in patients with dengue fever, including travelers.

Different approaches to characterize artificial breeding sites of Aedes aegypti using generalized linear mixed models.

BACKGROUND: As no globally accepted dengue vaccines or specific antiviral therapies are currently available, controlling breeding sites of Aedes aegypti is a target to prevent dengue outbreaks. The present study aimed to characterize outdoor artificial breeding sites in urban households using an exhaustive classification system. METHODS: A cross-sectional entomological survey was carried out in Colón city, Entre Ríos, Argentina, using a two-stage stratified sampling design during March and April 2014. The city was stratified given the degree of urbanization of each block, and blocks and households were randomly selected. All outdoor containers with water were inspected, and the presence of immature mosquitoes was recorded. Containers were classified according to physical, functional, and location attributes. Generalized linear mixed models were applied to take into account the aggregated nature of the data (containers in houses and houses in blocks). RESULTS: Overall, 207 houses were inspected. Out of 522 containers with water, 25% had immatures of Ae. aegypti (7336). In adjusted models, the abundance of immatures was higher in containers with increasing opening surface and volume, without roof cover, exposed to shadow, out of use or with functions related to gardening activities, household chores, water storage, or construction. At block level, immatures abundance was positively associated with the degree of urbanization. CONCLUSIONS: We detected high immatures abundance in containers associated with water utilization. This suggests that containers involved in these activities, whether directly (e.g., water storage) or indirectly (e.g., incomplete water drainage in the last use), are susceptible to present a high immature abundance. Although our results indicate the importance of the type of use over the type of container, we encourage the use of both classification criteria for artificial breeding sites of mosquitoes, mainly because these are complementary. Additionally, generalized linear mixed models allowed us to analyse predictor variables at different scales (container/house/block) and consider the lack of independence between observations. An exhaustive analysis of artificial breeding sites that use this analytical methodology can lead to new information that could help designing more appropriate tools for dengue surveillance and control.

Modeling the suppression dynamics of Aedes mosquitoes with mating inhomogeneity.

A novel strategy for controlling mosquito-borne diseases, such as dengue, malaria and Zika, involves releases of Wolbachia-infected mosquitoes as Wolbachia cause early embryo death when an infected male mates with an uninfected female. In this work, we introduce a delay differential equation model with mating inhomogeneity to discuss mosquito population suppression based on Wolbachia. Our analyses show that the wild mosquitoes could be eliminated if either the adult mortality rate exceeds the threshold [Formula: see text] or the release amount exceeds the threshold [Formula: see text] uniformly. We also present the nonlinear dependence of [Formula: see text] and [Formula: see text] on the parameters, respectively, as well as the effect of pesticide spraying on wild mosquitoes. Our simulations suggest that the releasing should be started at least 5 weeks before the peak dengue season, taking into account both the release amount and the suppression speed.

Effect of Larval Nutritional Regimes on Morphometry and Vectorial Capacity of Aedes aegypti for Dengue Transmission.

BACKGROUND: Nutritional level in larval diet of mosquito vectors influence on life history traits and vectorial capacity (VC). Therefore, the present study was carried out to assess the effect of larval diet concentration on vector bionomic and VC of Aedes aegypti in Sri Lanka. METHOD: Three batches of 400 Ae. aegypti larvae (first instar) were reared under different concentrations of larval diet (6%, 8%, and 10%; Volume/Volume), which was prepared by mixing 12.5 g of tuna meal, 9.0 g of bovine liver powder, and 3.5 g of Brewer's yeast, in 100 ml of distilled water. The effect of larval diet concentration on different morphometric and functional parameters of larvae (length and width of head, abdomen, survival rate, and pupation success), pupae (length and width of cephalothorax, survival rate, and adult emersion), adult (length and width of thorax, abdomen, survival rate, longevity, biting frequency and fecundity of adults) were examined. In addition, VC of Ae. aegypti was evaluated. The General Linear Model (GLM) was used for the statistical analysis. RESULTS: Larval head length, head width, thoracic width, abdominal length, abdominal width, total length, and survival rate significantly increased with higher doses of larval diet (P < 0.05). In case of pupae, length, and width of cephalothorax, survival rate and adult emergence rate denoted an increasing trend with the elevated larval diets. However, the variations of survival rate and adult emergence rate were statistically significant (P < 0.05). In adults, all morphometric parameters (thoracic length, abdominal length, abdominal width, and wing length) significantly increased with elevating larval diets levels (except for thoracic width) along with the biting frequency, fecundity, and survival rate (P < 0.05) of adult females. The VC also denoted significant variations (F 4,14 = 24.048; P < 0.05) with the larval diet concentration, whereby the highest VC of 196.37 was observed at 10% treatment. CONCLUSION: Larval food availability has a significant influence on the adult fitness and thus may affect the incidence of dengue due to variations in the VC of Ae.

Impact of adult mosquito control on dengue prevalence in a multi-patch setting: A case study in Kolkata (2014-2015).

Dengue is one of the deadliest mosquito-borne disease prevalent mainly in tropical and sub-tropical regions. Controlling the spread of this disease becomes a major concern to the public health authority. World Health Organization (WHO) adopted several mosquito control strategies to reduce the disease prevalence. In this work, a general multi-patch non-autonomous dengue model is formulated to capture the temporal and spatial transmission mechanism of the disease and the effectiveness of different adult mosquito control strategies in reducing dengue prevalence is evaluated. During the period (2014-2015) the dengue situation of Kolkata which is one of the most dengue affected city in India is considered in our study. Depending on geographical location, Kolkata is divided into five regions and our model is fitted to the monthly dengue cases of these five regions during the above-mentioned period. By considering control specific characteristics (e.g. efficacy, environment persistence) of the mosquito control strategies, we study the efficiency of three adult mosquito controls and their combined effect in reducing dengue prevalence. From our study, it is observed that control with higher environment persistence performs better in comparison to the controls having low environment persistence. It is also observed that, connectedness between the regions play a key role in the effectiveness of the control strategies.

Cartography of odor chemicals in the dengue vector mosquito (Aedes aegypti L., Diptera/Culicidae).

This study was aimed to identify the chemical compounds of Aedes aegypti that can be potentially used to develop pheromone-based vector control methods. In this study, we compared the chemical compounds collected from the organs of mosquitoes at different developmental stages in the life cycle. We also compared the composition and amount of extracts from the different tissues of male and female adult mosquito. Interestingly, we found large amount of C17-C20 ethyl and methyl esters in the wings of female and antennae of male mosquito. We also found that isopropyl esters, dodelactone, octadecenoic acid and medium-chain fatty acid increase drastically during the late larval stage (L4). Old adult mosquitoes showed remarkable increase in production of C16:1 and C18:1 methyl esters, as a first example of chemical signatures specifically associated with aging in the animals. This knowledge may open the ground to find new behaviorally-important molecules with the ability to control Aedes specifically.

Chagas Disease in Southern Coastal Ecuador: Coinfections with Arboviruses and a Comparison of Serological Assays for Chagas Disease Diagnosis.

Occurrence of Chagas disease and arbovirus coinfections is unknown, despite the vast co-endemic areas throughout the Americas. This study examined the proportion of individuals positive for Trypanosoma cruzi and coinfections with dengue, chikungunya, and Zika viruses in Machala, Ecuador (January 2014-December 2015). Chagas seropositivity was evaluated with five commercially available assays. Dengue infections were identified by nonstructural protein 1 rapid test and enzyme linked immunosorbent assay (ELISA), immunoglobulin M ELISA, and reverse transcription PCR (RT-PCR); chikungunya and Zika infections were identified by RT-PCR. Of 658 individuals, six were positive for T. cruzi (0.91%), including one T. cruzi/dengue coinfection and one T. cruzi/chikungunya/dengue coinfection. The clinical manifestations of coinfected individuals corresponded to severe dengue and dengue with warning signs, respectively. We observed discrepant results by using the Hemagen Chagas kit and the rapid test Chagas Detect Plus (false positives: 3.9% and 15.4%), highlighting the need to assess diagnostic assays in geographic regions with distinct taxonomic units of T. cruzi.

Characterization of Wolbachia enhancing domain in mosquitoes with imperfect maternal transmission.

A novel method to reduce the burden of dengue is to seed wild mosquitoes with Wolbachia-infected mosquitoes in dengue-endemic areas. Concerns in current mathematical models are to locate the Wolbachia introduction threshold. Our recent findings manifest that the threshold is highly dependent on the initial population size once Wolbachia infection alters the logistic control death rate of infected females. However, counting mosquitoes is beyond the realms of possibility. A plausible method is to monitor the infection frequency. We propose the concept of Wolbachia enhancing domain in which the infection frequency keeps increasing. A detailed description of the domain is presented. Our results suggest that both the initial population size and the infection frequency should be taken into account for optimal release strategies. Both Wolbachia fixation and extinction permit the oscillation of the infection frequency.

Risk factors for the presence of dengue vector mosquitoes, and determinants of their prevalence and larval site selection in Dhaka, Bangladesh.

Dengue viruses are responsible for over 100 million infections a year worldwide and are a public health concern in Bangladesh. Although risk of transmission is high, data on vector population characteristics are scanty in Bangladesh; therefore, a comprehensive prediction of the patterns of local virus transmission is not possible. Recognizing these gaps, multi-year entomological surveys were carried out in Dhaka, where the disease is most frequently reported. The specific objectives of the present study are threefold: i) to determine the risk factors for the presence of Aedes mosquitoes; ii) to identify the types of most productive and key containers; and iii) to estimate the effects of climatic factors on Aedes abundance in the city of Dhaka, Bangladesh. Entomological surveys were conducted in 12 out of 90 wards in Dhaka. These wards were selected using a probability proportional sampling procedure during the monsoon seasons in 2011, 2012 and 2013 and in the dry season in 2012. All containers inside and around sampled households were inspected for mosquito larvae and pupae, and containers were classified according to their relative size, use pattern, and materials of construction. During the study period (2011-2013), 12,680 larvae and pupae were collected. About 82% of the identified immature mosquitoes were Aedes aegypti, while the remainder were Ae. albopictus and other mosquito species. The largest number of immature mosquitoes was collected from tires and refrigerator trays during 2011 and 2012 monsoon seasons. Conversely, plastic drums were the most productive during the 2012 dry and 2013 monsoon season. Vehicle parts and discarded construction materials were the most efficient producers of Aedes mosquitoes in all surveys. The presence of Aedes mosquitoes was significantly (p < 0.05) higher in low socio-economic zones of Dhaka. Container location, presence of vegetation, and availability of shade for containers were also significantly associated with finding immature Aedes mosquitoes, based on multivariable analysis after confounder adjustment. Rainfall, temperature, and relative humidity also significantly affected the mean abundance of mosquitoes. Proper use, disposal, and recycling of the containers that effectively produce large numbers of Aedes vector mosquitoes may decrease the risk of arboviral transmission.

An assessment of false positive rates for malaria rapid diagnostic tests caused by non-Plasmodium infectious agents and immunological factors.

BACKGROUND: Malaria rapid diagnostic tests (RDTs) can produce false positive (FP) results in patients with human African trypanosomiasis and rheumatoid factor (RF), but specificity against other infectious agents and immunological factors is largely unknown. Low diagnostic specificity caused by cross-reactivity may lead to over-estimates of the number of malaria cases and over-use of antimalarial drugs, at the cost of not diagnosing and treating the true underlying condition. METHODS: Data from the WHO Malaria RDT Product Testing Programme was analysed to assess FP rates of 221 RDTs against four infectious agents (Chagas, dengue, Leishmaniasis and Schistosomiasis) and four immunological factors (anti-nuclear antibody, human anti-mouse antibody (HAMA), RF and rapid plasma regain). Only RDTs with a FP rate against clean negative samples less than 10% were included. Paired t-tests were used to compare product-specific FP rates on clean negative samples and samples containing non-Plasmodium infectious agents and immunological factors. RESULTS: Forty (18%) RDTs showed no FP results against any tested infectious agent or immunological factor. In the remaining RDTs significant and clinically relevant increases in FP rates were observed for samples containing HAMA and RF (P<0.001). There were significant correlations between product-matched FP rates for RF and HAMA on all RDT test bands (P<0.001), and FP rates for each infectious agent and immunological factor were also correlated between test bands of combination RDTs (P≤0.002). CONCLUSIONS: False positive results against non-Plasmodium infectious agents and immunological factors does not appear to be a universal property of malaria RDTs. However, since many malaria RDTs have elevated FP rates against HAMA and RF positive samples practitioners may need to consider the possibility of false positive results for malaria in patients with conditions that stimulate HAMA or RF.

Insecticide treated curtains and residual insecticide treatment to control Aedes aegypti: An acceptability study in Santiago de Cuba.

BACKGROUND: Within the context of a field trial conducted by the Cuban vector control program (AaCP), we assessed acceptability of insecticide-treated curtains (ITCs) and residual insecticide treatment (RIT) with deltamethrin by the community. We also assessed the potential influence of interviewees' risk perceptions for getting dengue and disease severity. METHODOLOGY/PRINCIPAL FINDINGS: We embedded a qualitative study using in-depth interviews in a cluster randomized trial (CRT) testing the effectiveness of ITCs and RIT in Santiago de Cuba. In-depth interviews (N = 38) were conducted four and twelve months after deployment of the tools with people who accepted the tools, who stopped using them and who did not accept the tools. Data analysis was deductive. Main reasons for accepting ITCs at the start of the trial were perceived efficacy and not being harmful to health. Constraints linked to manufacturer instructions were the main reason for not using ITCs. People stopped using the ITCs due to perceived allergy, toxicity and low efficacy. Few heads of households refused RIT despite the noting reasons for rejection, such as allergy, health hazard and toxicity. Positive opinions of the vector control program influenced acceptability of both tools. However, frequent insecticide fogging as part of routine AaCP vector control actions diminished perceived efficacy of both tools and, therefore, acceptability. Fifty percent of interviewees did feel at risk for getting dengue and considered dengue a severe disease. However, this did not appear to influence acceptability of ITCs or RIT. CONCLUSION/SIGNIFICANCE: Acceptability of ITCs and RIT was linked to acceptability of AaCP routine vector control activities. However, uptake and use were not always an indication of acceptability. Factors leading to acceptability may be best identified using qualitative methods, but more research is needed on the concept of acceptability and its measurement.

In vitro and in vivo thermal stress induces proliferation of Blastocystis sp.

Blastocystis sp. is known to be the most commonly found intestinal protozoan parasite in human fecal surveys and has been incriminated to cause diarrhea and abdominal bloating. Binary fission has been widely accepted as the plausible mode of reproduction for this parasite. The present study demonstrates that subjecting the parasites in vitro to higher temperature shows the proliferation of parasite numbers in cultures. Transmission electron microscopy was used to compare the morphology of Blastocystis sp. subtype 3 isolated from a dengue patient having high fever (in vivo thermal stress) and Blastocystis sp. 3 maintained at 41 °C (in vitro thermal stress) and 37 °C (control). Fluorescence stains like acridine orange (AO) and 4',6'-diamino-2-phenylindole (DAPI) were used to demonstrate the viability and nuclear content of the parasite for both the in vitro and in vivo thermal stress groups of parasites. Blastocystis sp. at 37 °C was found to be mostly vacuolar whereas the in vitro thermal stressed isolates at 41 °C were granular with electron dense material seen to protect the granules within the central body. Parasites of the in vivo thermal stressed group showed similar ultrastructure as the in vitro ones. AO and DAPI staining provided evidence that these granules are viable which develop into progenies of Blastocystis sp. These granular forms were then observed to rupture and release progenies from the mother cells whilst the peripheral cytoplasmic walls were seen to degrade. Upon exposure to high temperature both in vitro and in vivo, Blastocystis sp. in cultures show higher number of granular forms seen to be protected by the electron dense material within the central body possibly acting as a protective mechanism. This is possibly to ensure the ability to survive for the granules to be developed as viable progenies for release into the host system.

A community-based prospective cohort study of dengue viral infection in Malaysia: the study protocol.

BACKGROUND: Globally, dengue infections constitute a significant public health burden. In recent decades, Malaysia has become a dengue hyper-endemic country with the co-circulation of the four dengue virus serotypes. The cyclical dominance of sub-types contributes to a pattern of major outbreaks. The consequences can be observed in the rising incidence of reported dengue cases and dengue related deaths. Understanding the complex interaction of the dengue virus, its human hosts and the mosquito vectors at the community level may help develop strategies for addressing the problem. METHODS: A prospective cohort study will be conducted in Segamat district of Johor State in Peninsular Malaysia. Researchers received approval from the Malaysian Medical Research Ethics Committee and Monash University Human Research Ethics Committee. The study will be conducted at a Malaysian based health and demographic surveillance site over a 1 year period in three different settings (urban, semi-urban and rural). The study will recruit healthy adults (male and female) aged 18 years and over, from three ethnic groups (Malay, Chinese and Indian). The sample size calculated using the Fleiss method with continuity correction is 333. Sero-surveillance of participants will be undertaken to identify asymptomatic, otherwise healthy cases; cases with dengue fever who are managed as out-patients; and cases with dengue fever admitted to a hospital. A genetic analysis of the participants will be undertaken to determine whether there is a relationship between genetic predisposition and disease severity. A detailed medical history, past history of dengue infection, vaccination history against other flaviviruses such as Japanese encephalitis and Yellow fever, and the family history of dengue infection will also be collected. In addition, a mosquito surveillance will be carried out simultaneously in recruitment areas to determine the molecular taxonomy of circulating vectors. DISCUSSION: The research findings will estimate the burden of asymptomatic and symptomatic dengue at the community level. It will also examine the relationship between virus serotypes and host genotypes, and the association of the clinical manifestation of the early phase with the entire course of illness.

Species composition, co-occurrence, association and affinity indices of mosquito larvae (Diptera: Culicidae) in Mazandaran Province, northern Iran.

Although considerable progress has been made in the past years in management of mosquito borne diseases such as malaria, dengue, yellow fever and West Nile fever through research in biology and ecology of the vectors, these diseases are still major threats to human health. Therefore, more research is required for better management of the diseases. This investigation provides information on the composition, co-occurrence, association and affinity indices of mosquito larvae in Mazandaran Province, northern Iran. In a large scale field study, mosquito larvae were collected from 120 sentinel sites in 16 counties in Mazandaran Province, using standard 350 ml dipper. Sampling took place monthly from May to December 2014. Collected larvae were mounted on glass slides using de Faure's medium and were diagnosed using morphological characters. Totally, 19,840 larvae were collected including three genera and 16 species from 120 larval habitats, as follows: Anopheles claviger, Anopheles hyrcanus, Anopheles maculipennis s.l., Anopheles marteri, Anopheles plumbeus, Anopheles pseudopictus, Culex pipiens, Culex tritaeniorhynchus, Culex torrentium, Culex perexiguus, Culex territans, Culex mimeticus, Culex hortensis, Culiseta annulata, Culiseta longiareolata, and Culiseta morsitans. Predominant species were Cx. pipiens and An. maculipennis s.l. which show the highest co-occurrence. The pair of species An. hyrcanus/An. pseudopictus showed significant affinity and association. High co-occurrence of the predominant species Cx. pipiens and An. maculipennis s.l. in the study area is of considerable importance in terms of vector ecology. It was also revealed that An. pseudopictus/An. hyrcanus often occur sympatrically indicating their common habitat requirements. The information may be equally important when vector control measures are considered.

Forced egg retention and oviposition behavior of malaria, dengue and filariasis vectors to a topical repellent diethyl-phenylacetamide.

Egg retention and oviposition behavior of four species of mosquito vectors viz., Anopheles stephensi, Aedes aegypti, Aedes albopictus, and Culex quinquefasciatus to a topical insect repellent diethyl-phenylacetamide (DEPA) at 0.1-1000 mg/L was investigated under laboratory conditions. Based on oviposition activity indices, DEPA demonstrated concentration dependent oviposition deterrent effect to A. stephensi (-0.18 to -0.97), A. aegypti (-0.18 to -0.91) and A. albopictus (-0.50 to -0.98) females. In contrast, positive oviposition response by C. quinquefasciatus (+0.39 and +0.70) was observed respectively at 0.1 and 1 ppm, while 10 ppm of DEPA on water received 50% lesser egg rafts than control. Gravid Culex females laid no egg rafts at 100 and 1000 ppm DEPA treated bowls effecting 100% oviposition deterrence. Test mosquito females deposited most of their eggs (> 90%) in the absence of repellent odour, while DEPA odour on water surface forced them to retain huge numbers of eggs. Females of A. aegypti, A. albopictus and A. stephensi retained 49, 67 and 50% of total eggs, respectively throughout the experiment. Egg retention by Culex females due to DEPA on the water surface was ca. 65%, equivalent to 4 egg rafts. Therefore, DEPA at lower concentrations could effectively disturb the oviposition by these vectors. Application of repellents in small water bodies would help in reducing the population build up of mosquitoes near human households and could be useful in the integrated management of mosquito vectors.

Modelling releases of sterile mosquitoes with different strategies.

To prevent the transmissions of malaria, dengue fever, or other mosquito-borne diseases, one effective weapon is the sterile insect technique in which sterile mosquitoes are released to reduce or eradicate the wild mosquito population. To study the impact of the sterile insect technique on disease transmission, we formulate discrete-time mathematical models, based on difference equations, for the interactive dynamics of the wild and sterile mosquitoes, incorporating different strategies in releasing sterile mosquitoes. We investigate the model dynamics and compare the impact of the different release strategies. Numerical examples are given to demonstrate rich dynamical features of the models.

The larvicidal activity of Agave sisalana against L4 larvae of Aedes aegypti is mediated by internal necrosis and inhibition of nitric oxide production.

Dengue is a viral disease that affects about 50 million people per year around the world. The aim of this study was to investigate the larvicidal activity of Agave sisalana crude extract in order to develop a new insecticide against Aedes aegypti. In larvicidal activity assays, fourth-stage Ae. aegypti larvae were exposed to different concentrations of A. sisalana crude extract for 3, 6, 12, and 24 h for determining the LC50. Next, we explored its cytotoxic activity by flow cytometry. Furthermore, histological alterations were confirmed by histopathological analysis, and the nitric oxide (NO) production by hemocytes was checked after different periods of exposure to A. sisalana crude extract. The LC50 was 4.5 ± 0.07 mg/mL. In addition, flow cytometry revealed an increase of cellular necrosis (21 and 16.5 % after 12 and 24 h, respectively) in larvae that were exposed to A. sisalana crude extract. The histological analysis revealed cell lysis and destruction of the peritrophic membrane. Furthermore, there was a reduction in the concentration of NO in the hemolymph from larvae exposed to A. sisalana crude extract after 3, 6, and 24 h (5.3 ± 4.3 vs. 22.7 ± 5.2 µM, 4.3 ± 5.5 vs. 25.4 ± 6.6 µM, and 6 ± 1.7 vs. 37.1 ± 7.8 µM, respectively). Our findings show that A. sisalana crude extract constitutes an effective larvicidal agent against Ae. aegypti larvae due to its necrotizing activity in hemocytes and inhibition of the NO production.

Multi-scale analysis of the associations among egg, larval and pupal surveys and the presence and abundance of adult female Aedes aegypti (Stegomyia aegypti) in the city of Merida, Mexico.

Despite decades of research, there is still no agreement on which indices of Aedes aegypti (Stegomyia aegypti) (Diptera: Culicidae) presence and abundance better quantify entomological risk for dengue. This study reports the results of a multi-scale, cross-sectional entomological survey carried out in 1160 households in the city of Merida, Mexico to establish: (a) the correlation between levels of Ae. aegypti presence and abundance detected with aspirators and ovitraps; (b) which immature and egg indices correlate with the presence and abundance of Ae. aegypti females, and (c) the correlations amongst traditional Aedes indices and their modifications for pupae at the household level and within medium-sized geographic areas used for vector surveillance. Our analyses show that ovitrap positivity was significantly associated with indoor adult Ae. aegypti presence [odds ratio (OR) = 1.50; P = 0.03], that the presence of pupae is associated with adult presence at the household level (OR = 2.27; P = 0.001), that classic Aedes indices are informative only when they account for pupae, and that window screens provide a significant level of protection against peridomestic Ae. aegypti (OR = 0.59; P = 0.02). Results reinforce the potential of using both positive collections in outdoor ovitraps and the presence of pupae as sensitive indicators of indoor adult female presence.