Assessment of the Risk of Exotic Zika Virus Strain Transmission by Aedes aegypti and Culex quinquefasciatus from Senegal Compared to a Native Strain.

Zika virus (ZIKV) shows an enigmatic epidemiological profile in Africa. Despite its frequent detection in mosquitoes, few human cases have been reported. This could be due to the low infectious potential or low virulence of African ZIKV lineages. This study sought to assess the susceptibility of A. aegypti and C. quinquefasciatus to ZIKV strains from Senegal, Brazil, and New Caledonia. Vertical transmission was also investigated. Whole bodies, legs/wings and saliva samples were tested for ZIKV by real-time PCR to estimate infection, dissemination and transmission rates as well as the infection rate in the progeny of infected female A. aegypti. For A. aegypti, the Senegalese strain showed at 15 days post-exposure (dpe) a significantly higher infection rate (52.43%) than the Brazilian (10%) and New Caledonian (0%) strains. The Brazilian and Senegalese strains were disseminated but not detected in saliva. No A. aegypti offspring from females infected with Senegalese and Brazilian ZIKV strains tested positive. No infection was recorded for C. quinquefasciatus. We observed the incompetence of Senegalese A. aegypti to transmit ZIKV and the C. quinquefasciatus were completely refractory. The effect of freezing ZIKV had no significant impact on the vector competence of Aedes aegypti from Senegal, and vertical transmission was not reported in this study.

Trophic transfer and toxicity of heavy metals from dengue mosquito Aedes aegypti to predator dragonfly Tramea cophysa.

Heavy metal pollution in aquatic habitats can be detrimental to both prey and predators in a food web. To investigate the potential for bio-transfer and bioaccumulation of heavy metals between specific trophic levels, 3rd instar larvae of Aedes aegypti were exposed to mercury (Hg), lead (Pb), cadmium (Cd), copper (Cu), and zinc (Zn) for three consecutive generations and fed to dragonfly (Tramea cophysa) nymphs. Exposure to Hg caused the highest mortality in A. aegypti larvae and T. cophysa nymphs. Bioaccumulation and life-history parameters of A. aegypti, including egg hatching time, larval and pupal duration, male and female life span, and fecundity, were also evaluated after metals exposure. All life-history parameters except larval duration were significantly affected by heavy metal treatments. Bioaccumulation of metals in A. aegypti larvae and adults gradually and significantly increased from 1st to 3rd generation. To the best of our knowledge, this is the first study describing the acute toxicity of heavy metals to mosquitoes. Our study shows that heavy metals cause dietary toxicity to an aquatic predator, dragonfly, via trophic transfer, which could have considerable consequences on aquatic ecosystems.

Larvicidal and histopathological efficacy of inhabitant pathogenic bacterial strains to reduce the dengue vector competence.

BACKGROUND: Aedes aegypti is a primary vector of dengue virus, and the causative agent of dengue is emerging globally as one of the most important arboviral diseases currently threatening human populations. Therefore, vector control is presently the primary intervention method of population reduction, in which natural A. aegypti populations would be reduced with inhabitant bacterial strains that are unable to transmit dengue virus. RESULT: Based on the pathogenicity of strains, only four isolates effectively show larvicidal activity. The 16S rRNA gene sequences and the phylogeny depicted that the potential isolates were Bacillus firmus (MK791255), Bacillus paramycoides (MK788268), Bacillus siamensis (MK788212), and Bacillus licheniformis (MK791256). After 24 and 48 hours exposure, the B. licheniformis strain (cell mass of 2.2 × 107 CFU mL-1 ) showed potent larvicidal activity with LC50 of 16.22 µg mL-1 and 9.57 µg mL-1 and the B. paramycoides (cell mass of 3.1 × 107 CFU mL-1 ) strain inhibits the larval and pupal development with LC50 of 42.62 µg mL-1 and 26.97 µg mL-1 . Intermittent stages and causes of abscess in the gut and siphon regions were observed through histopathological studies. These two bacterial strains extend larval duration up to 15-16 days as well as reduce development. CONCLUSION: These studies demonstrate the challenge for dengue vector in reducing developmental and reproduction competence. © 2020 Society of Chemical Industry.

A single unidirectional piRNA cluster similar to the flamenco locus is the major source of EVE-derived transcription and small RNAs in Aedes aegypti mosquitoes.

Endogenous viral elements (EVEs) are found in many eukaryotic genomes. Despite considerable knowledge about genomic elements such as transposons (TEs) and retroviruses, we still lack information about nonretroviral EVEs. Aedes aegypti mosquitoes have a highly repetitive genome that is covered with EVEs. Here, we identified 129 nonretroviral EVEs in the AaegL5 version of the A. aegypti genome. These EVEs were significantly associated with TEs and preferentially located in repeat-rich clusters within intergenic regions. Genome-wide transcriptome analysis showed that most EVEs generated transcripts although only around 1.4% were sense RNAs. The majority of EVE transcription was antisense and correlated with the generation of EVE-derived small RNAs. A single genomic cluster of EVEs located in a 143 kb repetitive region in chromosome 2 contributed with 42% of antisense transcription and 45% of small RNAs derived from viral elements. This region was enriched for TE-EVE hybrids organized in the same coding strand. These generated a single long antisense transcript that correlated with the generation of phased primary PIWI-interacting RNAs (piRNAs). The putative promoter of this region had a conserved binding site for the transcription factor Cubitus interruptus, a key regulator of the flamenco locus in Drosophila melanogaster Here, we have identified a single unidirectional piRNA cluster in the A. aegypti genome that is the major source of EVE transcription fueling the generation of antisense small RNAs in mosquitoes. We propose that this region is a flamenco-like locus in A. aegypti due to its relatedness to the major unidirectional piRNA cluster in Drosophila melanogaster.

Geometric morphometrics of Aedes aegypti populations and study of transmission of arboviral diseases in Barreiras, Brazil

Aedes (Stegomyia) aegypti (A. aegypti) transmits arboviral diseases of high public health importance, including those caused by Zika virus (ZIKV), Dengue virus (DENV), Chikungunya virus (CHIKV) and Yellow fever virus (YFV). Barreiras is a city with 157,638 inhabitants in the West of the State of Bahia, Northeast of Brazil. The climate is dry, with well-determined and concentrated seasons of rains. The city is crossed by a Federal Highway and by the Rio Grande river. In this study, we aimed to understand the dynamics of mosquito vectors and arboviral diseases in Barreiras. We used correlation statistics to investigate a possible relationship among rains, mosquito abundance and transmission of diseases. In addition, as a preliminary population genetics estimate, we used geometric morphometrics to compare mosquitoes from areas limited by a highway and a river. We found that i) infestation occurs in rain-dependent cycles and that ii) both, the river and the highway segregate populations of A. aegypti in different areas of the studied city. Our results indicate that it is necessary to treat anthropic containers with mosquito breading capacity during both, the dry and rain seasons in urban areas similar to Barreiras.

Geometric morphometrics of Aedes aegypti populations and study of transmission of arboviral diseases in Barreiras, Brazil

Abstract Aedes (Stegomyia) aegypti (A. aegypti) transmits arboviral diseases of high public health importance, including those caused by Zika virus (ZIKV), Dengue virus (DENV), Chikungunya virus (CHIKV) and Yellow fever virus (YFV). Barreiras is a city with 157,638 inhabitants in the West of the State of Bahia, Northeast of Brazil. The climate is dry, with well-determined and concentrated seasons of rains. The city is crossed by a Federal Highway and by the Rio Grande river. In this study, we aimed to understand the dynamics of mosquito vectors and arboviral diseases in Barreiras. We used correlation statistics to investigate a possible relationship among rains, mosquito abundance and transmission of diseases. In addition, as a preliminary population genetics estimate, we used geometric morphometrics to compare mosquitoes from areas limited by a highway and a river. We found that i) infestation occurs in rain-dependent cycles and that ii) both, the river and the highway segregate populations of A. aegypti in different areas of the studied city. Our results indicate that it is necessary to treat anthropic containers with mosquito breading capacity during both, the dry and rain seasons in urban areas similar to Barreiras.

Similar Neural Pathways Control Foraging in Mosquitoes and Worms.

Female Aedes aegypti mosquitoes bite human hosts to obtain a blood meal and, in the process, act as vectors for many disease-causing viruses, including the dengue, chikungunya, yellow fever, and Zika viruses. After a complete meal, the female mosquitoes lose attraction to their hosts for several days. New research shows that pharmacological activation of neuropeptide Y-like receptor (NPYLR) signaling elicits host aversion in female mosquitoes. This behavior of mosquitoes shows remarkable similarities to a bacterial-aversion behavior of the nematode Caenorhabditis elegans Feeding on pathogenic bacteria causes bloating of the gut in C. elegans that leads to activation of NPYLR signaling and bacterial aversion. Several studies suggest that this newly discovered mechanism underlying foraging may be conserved across a large number of species. A better understanding of the regulation of NPYLR signaling pathways could provide molecular targets for the control of eating behaviors in different animals, including human-disease vectors.

Dengue virus infection induces chromatin remodeling at locus AAEL006536 in the midgut of Aedes aegypti.

OBJECTIVE: To identify and characterize Aedes aegypti's AAEL006536 gene proximal upstream cis-regulatory sequences activated by dengue virus infection. MATERIALS AND METHODS: A. aegypti Rockefeller strain mosquitoes were blood fed or infected with dengue virus 2. Open chromatinprofiling was then carried out in pools of midguts from each group of mosquitoes. RESULTS: The proximal upstream region does not contain open chromatin sites in the midguts of blood-fed mosquitoes as detected by FAIRE-qPCR. In contrast, two cis-regulatory sites were identified in the same upstream region of dengue virus-infected mosquito midguts. The distal sequence contains STAT-, REL- and C/EBP-type transcription factor binding sites. CONCLUSIONS: The activation of two proximal cis-regulatory sequences, induced by dengue virus infection, is mediated by chromatin remodeling mechanisms. Binding sites suggest a dengue virus infectioninduced participation of immunity transcription factors in the up-regulation of this gene. This suggests the participation of the AAEL006536 gene in the mosquito's antiviral innate immune response.

Dengue virus infection induces chromatin remodeling at locus AAEL006536 in the midgut of Aedes aegypti / La infección por virus dengue induce remodelación de la cromatina en el locus AAEL006536 en el intestino medio de Aedes aegypti

Abstract: Objective: To identify and characterize Aedes aegypti's AAEL006536 gene proximal upstream cis-regulatory sequences activated by dengue virus infection. Materials and methods: A. aegypti Rockefeller strain mosquitoes were blood fed or infected with dengue virus 2. Open chromatin profiling was then carried out in pools of midguts from each group of mosquitoes. Results: The proximal upstream region does not contain open chromatin sites in the midguts of blood-fed mosquitoes as detected by FAIRE-qPCR. In contrast, two cis-regulatory sites were identified in the same upstream region of dengue virus-infected mosquito midguts. The distal sequence contains STAT-, REL- and C/EBP-type transcription factor binding sites. Conclusion: The activation of two proximal cis-regulatory sequences, induced by dengue virus infection, is mediated by chromatin remodeling mechanisms. Binding sites suggest a dengue virus infection-induced participation of immunity transcription factors in the up-regulation of this gene. This suggests the participation of the AAEL006536 gene in the mosquito's antiviral innate immune response.

Resumen: Objetivo: Identificar y caracterizar las secuencias reguladoras activadas por la infección por virus dengue en la región proximal del gen AAEL006536 de Aedes aegypti. Material y métodos: Mosquitos de la cepa Rockefeller de A. aegypti se infectaron con virus dengue o se alimentaron con sangre. Se obtuvieron los perfiles de cromatina abierta del locus en los intestinos de cada uno de los grupos. Resultados: Se identificaron dos sitios reguladores solo en los intestinos de mosquitos infectados por virus dengue. El sitio distal contiene sitios de unión a factores de transcripción tipo REL, STAT y C/EBP. Conclusiones: La activación de dos sitios reguladores proximales está mediada por la remodelación de la cromatina. Los sitios de unión a factores de transcripción en el sitio regulador distal sugieren la participación de las vías de inmunidad en la regulación del gen. Esto sugiere la participación de este gen en la respuesta inmune del mosquito frente a la infección viral.

Larvicidal and ovicidal activity of Terminalia chebula Retz. (Family: Combretaceae) medicinal plant extracts against Anopheles stephensi, Aedes aegypti and Culex quinquefasciatus.

Insect-borne diseases remain to this day a major source of illness and death worldwide. The resistance to chemical insecticides among mosquito species has been considered as a setback in vector control. Mosquito control programs, botanical origin may have the potential to eliminate eggs and larvae. So, the larvicidal and ovicidal activities of crude benzene, hexane, ethyl acetate, chloroform and methanol extracts of Terminalia chebula were assayed for their toxicity against three important vector mosquitoes, viz., Anopheles stephensi, Aedes aegypti, and Culex quinquefasciatus. The larval mortality was observed after 24 h of exposure. All extracts showed moderate larvicidal effects; however, the highest larval mortality was found in the methanol extract of T. chebula against the larvae of A. stephensi, A. aegypti, and C. quinquefasciatus with the LC50 values were 87.13, 93.24 and 111.98 ppm, respectively. Mean percent hatchability of the ovicidal activity was observed 48 h post treatment. The percent hatchability was inversely proportional to the concentration of extract and directly proportional to the eggs. All the five solvent extracts showed moderate ovicidal activity; however, the maximum egg mortality (zero hatchability) was observed in the methanol extract of T. chebula at 200 and 250 ppm against A. stephensi, A. aegypti and C. quinquefasciatus showed 100% mortality at 300 ppm. No mortality was recorded in the control. The finding of the present investigation revealed that the leaf extract of Terminalia chebula possesses remarkable larvicidal and ovicidal activity against medically important vector mosquitoes and make this plant product promising as an alternative to synthetic insecticide in mosquito control programs.

Modelling the association of dengue fever cases with temperature and relative humidity in Jeddah, Saudi Arabia-A generalised linear model with break-point analysis.

The aim of this study was to examine the role of environmental factors in the temporal distribution of dengue fever in Jeddah, Saudi Arabia. The relationship between dengue fever cases and climatic factors such as relative humidity and temperature was investigated during 2006-2009 to determine whether there is any relationship between dengue fever cases and climatic parameters in Jeddah City, Saudi Arabia. A generalised linear model (GLM) with a break-point was used to determine how different levels of temperature and relative humidity affected the distribution of the number of cases of dengue fever. Break-point analysis was performed to modelled the effect before and after a break-point (change point) in the explanatory parameters under various scenarios. Akaike information criterion (AIC) and cross validation (CV) were used to assess the performance of the models. The results showed that maximum temperature and mean relative humidity are most probably the better predictors of the number of dengue fever cases in Jeddah. In this study three scenarios were modelled: no time lag, 1-week lag and 2-weeks lag. Among these scenarios, the 1-week lag model using mean relative humidity as an explanatory variable showed better performance. This study showed a clear relationship between the meteorological variables and the number of dengue fever cases in Jeddah. The results also demonstrated that meteorological variables can be successfully used to estimate the number of dengue fever cases for a given period of time. Break-point analysis provides further insight into the association between meteorological parameters and dengue fever cases by dividing the meteorological parameters into certain break-points.

Potential for Extrinsic Incubation Temperature to Alter Interplay Between Transmission Potential and Mortality of Dengue-Infected Aedes aegypti.

The extrinsic incubation period is a critical component in the assessment of arboviral transmission potential. It defines the time it takes for a mosquito to become infectious following exposure to an arbovirus. Since this is a temporal process, the lifespan of a mosquito is intimately tied to the extrinsic incubation period and thus transmission potential of these viruses. Temperature is a known effector of both vector competence (the ability of a vector to transmit a pathogen) and mosquito mortality, but the interaction among temperature, vector competence, and mosquito mortality is not well characterized. Herein, we investigate this interaction for dengue virus, serotype 2, and its primary vector Aedes aegypti where we found that at 30 °C, infection and/or dissemination shortened the average lifespan of the mosquito and that when considering only mosquitoes with a disseminated infection, those incubated at 26 °C lived significantly longer.

Evidence for natural vertical transmission of chikungunya viruses in field populations of Aedes aegypti in Delhi and Haryana states in India-a preliminary report.

Aedes aegypti and Aedes albopictus are principal vectors for the transmission of chikungunya virus (CHIKV). India is a hub for both dengue and chikungunya infections and there are several reports of co-infection of dengue and chikungunya virus in the clinical scenario. The present pilot entomological survey was conducted to evaluate vertical transmission of CHIKV in Aedes field populations. Aedes immature (larvae and pupae) collection was done in 2012, over a period of six months from selected sites in Delhi and Haryana, India. The immatures collected were reared for adult emergence and species identification was done. A. aegypti male and female mosquitoes were separated and pooled collection spot-wise, RNA extracted and RT PCR performed to test for the presence of CHIKV in the pools. Container index (CI) and minimum infection rate (MIR) were estimated. From study areas that tested positive for CHIKV, adult collections were made and females upon feeding on uninfected blood in laboratory were allowed to lay eggs. The progeny that emerged from these field-collected mothers were tested for CHIKV presence. Our pilot survey showed the existence of A. aegypti population even during peak summer season in a few foci which eventually helped the mosquitoes to tide over adverse environmental conditions and with the start of rainfall, the population exploded within a short period of time. Immatures collected from field and progeny of adults collected from the field were CHIKV positive demonstrating the presence of vertical transmission of chikungunya virus in field population of A. aegypti. The present study further demonstrates the importance of identifying permanent breeding sites for proper Aedes species control.

Empirical evidence of the effect of school gathering on the dynamics of dengue epidemics.

INTRODUCTION: Dengue fever is an important vector-transmitted disease that affects more than 100 countries worldwide. Locations where individuals tend to gather may play an important role in disease transmission in the presence of the vector. By controlling mosquitoes' breeding places, this study aims to analyze the effect of reducing transmission in elementary schools (grades 1-9) on the dynamics of the epidemic at a regional level. MATERIALS AND METHODS: In 2007, we implemented a massive campaign in a region of México (Colima state, 5,191 km(2), population 568,000) focused on training janitors to locate and avoid mosquitoes' breeding places, the objective being to maintain elementary schools free of mosquitoes. RESULTS: We observed 45% reduction in dengue incidence compared to the previous year. In contrast, the rest of Mexico observed an 81% increase in incidence on average. DISCUSSION: Costs associated with campaigns focusing on cleaning schools are very low and results seem to be promising. Nevertheless, more controlled studies are needed.

Is transfusion-transmitted dengue fever a potential public health threat?

Dengue is an arboviruses due to single-stranded enveloped ribonucleic acid viruses, named dengue viruses (DENV), that include four serotypes and are mainly transmitted via the bite of mosquitoes of the genus Aedes (A. aegypti and A. albopictus). The distribution of the disease was historically limited to intertropical areas; however, during the last thirty years, the perimeter of the disease extended considerably and temperate areas are now at risk of outbreaks. The present global burden of dengue is considerable: 2.5 billion people over more than 100 countries are concerned; 50 to 100 million infections occur every year, with a number of fatal cases of approximately 20000. Although frequently asymptomatic or limited to a mild fever, dengue is responsible for severe cases mainly consecutive to the occurrence of hemorrhagic complications that can lead to shock and death, notably in children from poor-resource settings. The place of DENV as a transfusion-transmitted pathogen has been recognized only in 2008. At the present time, only five cases of transfusion-transmitted dengue, including one case of dengue hemorrhagic fever, have been formerly documented. This review provides a general overview of dengue, its viruses and their vectors. It replaces the disease in the context of other viral diseases transmitted by arthropods. It discusses the threat of dengue on the supply of blood products in endemic and non endemic areas. Finally, it describes the specific and non specific measures available for improving the security of blood products with regards to this emerging risk. Interestingly, in 2009, the American Association of Blood Banks placed DENV in the highest category of emerging infectious agents for their potential impact on transfusion recipient safety for the next years in North America.

Reduced competitiveness of Wolbachia infected Aedes aegypti larvae in intra- and inter-specific immature interactions.

Wolbachia are maternally inherited intracellular bacteria that frequently infect a diverse range of arthropod species. Empirical and theoretical studies examining Wolbachia invasiveness have emphasized Wolbachia effects on adult hosts, but recent studies show that Wolbachia impacts on immature hosts can be important also. Here, we have examined for effects of Wolbachia infection in Aedes aegypti. Specifically, differential survivorship is observed when young larvae (1st instar) are exposed to older Aedes albopictus larvae (4th instar) or con-specific larvae. In an additional experiment, we have examined for differential behavior and observed that Wolbachia-infected larvae differ from uninfected larvae in their reaction to light stimulation. Our results support a hypothesized effect of Wolbachia on A. aegypti larval behavior. The results are discussed in relation to the ability of Wolbachia to invade natural populations and recently applied public health strategies that target the replacement or suppression of this important disease vector.