Development and ageing of the salivary glands of adult male Aedes aegypti (L.) and Aedes togoi (theobald) mosquitoes (Dipteria : Culcidae)

The salivary glands of adult male Aedes aegypti and Aedes togoi (Diptera : Culicidae), varying in age from less than 1 day after emergence to 42 days or 33 days respectively, were examined by light microscopy. Following emergence, the trilobed glands rapidly accumulate secretory product and attain their full size within about 48 hr. The amount of secretion in fully developed glands shows marked individual variation, but it is predominantly located in the posterior parts of the glands. Ageing changes begin to appear after about 8 days (A. aegypti) or 10–11 days (A. togoi), and although there is wide individual variation in their extent, in general, they become progressively more severe until at the age of 4–5 weeks all mosquitoes are substantially affected. Since other tissues, including ganglia, also regress with age, it is unlikely that salivary gland changes are the prime cause of death.     

Cell replication in the follicular epithelium of the adult mosquito

Females of the mosquitoes Culex pipiens molestus, Aedes togoi, Mansonia uniformis, and Aedes aegypti show active mitosis and an increase in cell number in the follicular epithelium of the ovary immediately after emergence from the pupa. Development up to stage III in the autogenous species, when mitosis in the follicular epithelium ceases, depends upon the continuity of cell proliferation over the first 48 hr after emergence (C. p. molestus, A. togoi) and on the number of cells originally in the follicle at emergence (A. togoi). The follicular epithelium in the anautogenous species ceases to divide in Mansonia, or the rate of division is considerably reduced in A. aegypti, 24–48 hr after emergence. Mitosis in the follicular epithelium of A. aegypti is renewed after the blood meal. A number of cells are released from phase G2 of the mitotic cycle into mitosis and other cells are initiated into DNA synthesis within 4 hr of the mosquito feeding on blood.     

Laboratory culture of Aedes (Finlaya) togoi (Theobald), 1907 and measurements of its susceptibility to insecticides

The literature on Aedes togoi is reviewed. This oriental mosquito occasionally acts as a vector for filarial or virus diseases and is sometimes a nuisance from its bites. Eggs from Taiwan (Formosa) were brought to London in 1958. A laboratory colony was reared from these eggs and maintained by methods similar to those commonly used for Aedes aegypti. Various aspects of the bionomics were studied, including the lengths of stages, mortality during development and the effects of copulation, feeding and salt content of water on oviposition. The levels of susceptibility to DDT, BHC and dieldrin were measured by the World Health Organisation (W.H.O.) method and compared with those of colonies of A. aegypti originating in Taiwan and the Pescadores.

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Zusammenfassung Eine Laborzucht von Aedes togoi wurde erfolgreich eingerichtet und die Kulturtechnik beschrieben. Die Ergebnisse der vorläufigen Beobachtungen über ihre Biologie werden ebenfalls mitgeteilt. Die Kulturtechnik unterscheidet sich nicht sehr von den gewöhnlich für Aedes aegypti angewandten. Jedoch wurde die Dauer eines Generationszyklus viel länger als die von Aedes aegypti befunden. Bei einem Teil der Weibchenpopulation konnte Autogenie beobachtet werden. Kopulationen wurden nicht in allen benutzten Käfigtypen durch Augenbeobachtung festgestellt, durch Sektion der Weibchen jedoch in allen Käfigformen bis zu den kleinsten von 19×19×19 cm nachgewiesen. Unbegattete Weibchen legen nachweislich ebenso gleichmä£ig Eier ab wie begattete, aber die produzierten Eier waren alle steril. Die von befruchteten autogenen Weibchen abgelegten Eier waren lebensfähig und entwickelten sich zu normalen Imagines. Erwachsene Weibchen, denen 2-und 5%ige Salzlösungen, 16%ige Zuckerlösung und frisches Leitungswasser geboten wurden, vermieden Eiablagen in die Gläser mit Salzlösungen, obwohl die Larven der Stammzucht in brackigen Felsentümpeln lebend gefunden wurden. In Hongkong wurden die Larven dieser Art in Gewässern mit einer Salinität von 0,046 bis 1,85% Chlor gefunden, annähernd dem höchsten Prozentgehalt in Seewasser. In der vorliegenden Untersuchung ertrugen die Larven Salzlösungen bis zu 5% und die Eier entwickelten sich ganz normal zu Erwachsenen in Salzlösungen von 1, 2, 3, 4 und 5%. Die Entwicklung wurde in höheren Konzentrationen nur schwach verzögert befunden. Es bestanden keine signifikanten Unterschiede zwischen den Sterblichkeitsraten bei verschiedenen Konzentrationen, nur in der höchsten Konzentration (5% Salzlösung) zeigte sich eine höhere Mortalität.Die Empfindlichkeit von Aedes togoi und zwei Stämmen von Aedes aegypti wurde mit den W.H.O.-Prüfwannen gemessen. Die Imagines wurden mit DDT und die Larven mit DDT, Dieldrin und gamma-BHC geprüft. Der vorliegende Stamm von Aedes togoi entstammte einem Gebiet, das von überständigem Antimalaria-DDT-Gebäudesprückstand bedeckt war; so konnte die Möglichkeit einer Resistenzentwicklung bei diesen Mücken erwartet werden. Unglücklicherweise ist kein Stamm von Aedes togoi verfügbar von Orten, die von DDT-Begiftung frei sind; deshalb kann seine mögliche Resistenz genen DDT nicht abgeschätzt werden. Jedoch erwies er sich als ebenso empfindlich wie normale Stämme von Aedes aegypti aus dem südlichen Taiwan und Pescadores. Die männlichen Imagines von Aedes togoi sind viel stärker DDT-empfindlich als die Weibchen und beide Geschlechter normaler Stämme von Aedes aegypti. Im Larvalstadium ist Aedes togoi viel toleranter gegen DDT als normale Stämme von Aedes aegypti. Die Resulte sind in 12 Tabellen und in den Figuren 1 und 2 aufgeführt.

     

Genetics of glucosephosphate isomerase in Aedes togoi (Diptera: Culicidae)

The genetics of glucosephosphate isomerase (E.C. 5.3.1.9) in two strains (Malaysian and Taiwan) of Aedes togoi is reported. Three electrophoretic phenotypes were present in both sexes. The zymogram patterns were identical in both strains of A. togoi. The phenotypes were governed by a pair of codominant alleles. The allele frequency of the slow-moving band was 0.63 in the Malaysian strain and was 0.86 and 0.82 in F161 and F169 generations, respectively, of the Taiwan strain. The sample studied was in good accord with Hardy-Weinberg expectations.     

Physiological effects of carbohydrates on survival, metabolism, and flight potential of female Aedes taeniorphynchus

Starved Aedes taeniorhynchus and Aedes aegypti females 2 days after emergence were fed ad lib. a mixture of 10% test carbohydrate solution and 5% sorbose solution (as a phagostimulant) and maintained for survival. Also, starved A. taeniorhynchus females 5 days after emergence were fed, with a micro-pipette, 2 μl/female of a mixture of 20% test carbohydrate and 5% sorbose. These females were analysed for glycogen accumulation 18 hr after feeding, and they were flown tethered on a flight mill immediately as well as 24 hr after feeding. Twenty-eight carbohydrates including sugar alcohols were tested.     

Structural organization of the brain and subesophageal ganglion of male Aedes aegypti (L.) (Diptera : Culicidae)

The brain and subesophageal ganglion of male Aedes aegypti (L.) (Diptera : Culicidae) are described from cryofractures and silver-stained, semithin (0.5 μm) serial sections of whole heads observed in the scanning and light microscopes. The brain and subesophageal ganglion of male A. aegypti are fused. The major structures of the brain include the protocerebral lobes and bridge, the mushroom bodies, central complex of the protocerebrum, the mechanosensory regions and olfactory loves of the deutocerebrum, and the tritocerebrum. Major commissures of the brain are the anterior optic tract, central commissure, posterior dorsal commissure, and subesophageal commissure. The structural associations of brain components with each other and the subesophageal ganglion, as well as the paths of the major nerve tracts in male A. aegypti are described and compared with those in other Diptera.     

Peripheral cells in the salivary glands of female Aedes aegypti and A. togoi mosquitoes

A new type of cell, the peripheral cell, is described. These cells are located at the perimeter of the simple tubules which form the distal zones of the lateral lobes of the salivary glands of female Aedes aegypti and A. togoi. They may represent degenerate secretory cells which are segregated so that their altered secretory product cannot be discharged during blood feeding.     

Morphological Changes in the Midgut of Aedes aegypti L. (Diptera: Culicidae) Larvae Following Exposure to an Annona coriacea (Magnoliales: Annonaceae) Extract

Bioinsecticides are important in the control of disease vectors, but data regarding their physiological effects on target insects are incomplete. This study describes morphological changes that occur in the midgut of third instar Aedes aegypti L. (Diptera: Culicidae) following treatment with a methanolic extract of Annona coriacea (Magnoliales: Annonaceae). Dissected midguts were subdivided into anterior and posterior regions and analyzed by light and scanning electron microscopy. Insects exposed to the extract displayed intense, destructive cytoplasmic vacuolization in columnar and regenerative midgut cells. The apical surfaces of columnar cells exhibited cytoplasmic protrusions oriented toward the lumen, suggesting that these cells could be involved in apocrine secretory processes and/or apoptosis. We report that A. coriacea extracts induced morphological alterations in the midgut of A. aegypti midgut larvae, supporting the use of plant extracts for control of the dengue vector.     

Modelling distributions of Aedes aegypti and Aedes albopictus using climate,host density and interspecies competition

Florida faces the challenge of repeated introduction and autochthonous transmission of arboviruses transmitted by Aedes aegypti and Aedes albopictus. Empirically-based predictive models of the spatial distribution of these species would aid surveillance and vector control efforts. To predict the occurrence and abundance of these species, we fit a mixed-effects zero-inflated negative binomial regression to a mosquito surveillance dataset with records from more than 200,000 trap days, representative of 53% of the land area and ranging from 2004 to 2018 in Florida. We found an asymmetrical competitive interaction between adult populations of Aedes aegypti and Aedes albopictus for the sampled sites. Wind speed was negatively associated with the occurrence and abundance of both vectors. Our model predictions show high accuracy (72.9% to 94.5%) in validation tests leaving out a random 10% subset of sites and data since 2017, suggesting a potential for predicting the distribution of the two Aedes vectors.     

Histological changes in the Dengue vector,Aedes aegypti (Diptera: Culicidae) larvae treated with neem oil loaded niosomes

Mosquitoes are one of the greatest threats to human health around the globe. They act as vectors for common diseases like Malaria, Dengue, Chikungunya, etc. Niosomes encapsulated with neem oil showed a significant mortality rate against Aedes aegypti larvae when treated for 24 h. In this study, the histological changes that led to the mortality of the Aedes aegypti mosquito larvae were studied. Organs of the late III-instar stage larvae such as head, optic lobes, cuticle, adipose tissue, midgut region, haemolymph were investigated. Several histological alterations such as disorientation of the brain and antenna in the head part, damage in the optic lobe and microvilli were observed. Total disruption was seen in the inner and outer retractor muscles of the larval body. The midgut and hindgut regions were disintegrated due to the damage to the fat bodies in the region. A Series of such histological changes in the body of mosquito larvae compared to the control larvae hindered metabolic functions leading to death. The results suggested that the neem oil loaded niosomes could be used as a biocontrol agent against the Dengue vector, Aedes aegypti larvae.     

Identification of salivary proteins in Aedes togoi (Theobald) that bind to mouse immunoglobulin E

Mosquito saliva contains a number of immunogenic molecules that can induce allergic reactions in host animals. The oriental tiger mosquito, Aedes togoi (Theobald), feeds on both humans and animals and is also a vector of various pathogenic viruses. However, information on immunogenic antigens in the saliva of this mosquito species is scarce. In this study, we used immunohistochemical staining and confocal microscopy to determine the fine structure of the acini lobes of salivary glands of A. togoi. In addition, we identified proteins from salivary gland extracts that reacted to mouse immunoglobulin E in an immunoblot analysis. At least nine proteins, ranging in size from 10 to 145 kDa, bound to IgEs; a 33 and 57.5 kDa protein showed the strongest binding signal. Our results provide important information on the identities of potential allergens in mosquito saliva.     

New traps for the capture of Aedes aegypti (Linnaeus) and Aedes albopictus (Skuse) (Diptera: Culicidae) eggs and adults

The control of arboviruses carried by Aedes aegypti (Linnaeus) and Aedes albopictus (Skuse) can be performed with tools that monitor and reduce the circulation of these vectors. Therefore, the efficiency of four types of traps in capturing A. aegypti and A. albopictus eggs and adults, with the biological product Vectobac WG, was evaluated in the field. For this, 20 traps were installed in two locations, which were in the South (Londrina, Paraná) and North (Manaus, Amazonas) Regions of Brazil, from March to April 2017 and January to February 2018, respectively. The UELtrap-E (standard trap) and UELtrap-EA traps captured A. aegypti and A. albopictus eggs: 1703/1866 eggs in Londrina, and 10268/2149 eggs in Manaus, respectively, and presented high ovitraps positivity index (OPI) values (averages: 100%/100% in Londrina, and 100%/96% in Manaus, respectively); and high egg density index (EDI) values (averages: 68/75 in Londrina, and 411/89 in Manaus, respectively), so they had statistically superior efficiency to that of the CRtrap-E and CRtrap-EA traps in both regions, that captured less eggs and adults: 96/69 eggs in Londrina, and 1091/510 eggs in Manaus, respectively. Also presented lower OPI values (averages: 28%/4% in Londrina, and 88%/60% in Manaus, respectively); and lower EDI values (averages: 10.5/9 in Londrina, and 47/30 in Manaus, respectively). The capture ratios of Aedes adults in the UELtrap-EA and CRtrap-EA traps in Londrina and Manaus were 53.3%/29.5% and 0%/9.8%, respectively. UELtrap-EA can be adopted as efficient tool for Aedes monitoring due to their high sensitivity, low cost and ease of use.     

Germination of Bacillus thuringiensis var. israelensis spores in the gut of Aedes larvae (Diptera: Culicidae)

In laboratory experiments, germination and growth of Bacillus thuringiensis israelensis in the gut of Aedes aegypti and A. vexans larvae (Culicidae: Diptera) was observed. The number of spores and vegetative cells in the gut of living larvae and in cadavers was estimated by plaing homogenized larvae on selective agar plates. The number of spores per gut increased in the first 40–140 min of exposure to a maximum, and decreased in the subsequent time, demonstrating spore germination in living larvae, moribunds, and in cadavers. Twenty-four hours after the death of the larvae, a minimal amount of spores, but an increased number of vegetative cells, was found in cadavers. In A. aegypti larvae, germination and growth of B. thuringiensis israelensis in the larval gut was photographically documented.     

Chikungunya Virus and Aedes Mosquitoes: Saliva Is Infectious as soon as Two Days after Oral Infection

Background

Aedes aegypti and Aedes albopictus are potential vectors of chikungunya virus (CHIKV). The recent CHIKV outbreaks were caused by a new variant characterized by a mutation in the E1 glycoprotein gene (E1-226V) which has favored a better transmissibility by Ae. albopictus. As Ae. albopictus tends to replace Ae. aegypti in many regions, one question remained: is Ae. albopictus as efficient as Ae. aegypti to transmit the variant E1-226V of CHIKV?

Methodology and Findings

We infected orally both species with the variant E1-226V and estimated the infection, the viral dissemination, and the transmission rate by real time RT-PCR. Additionally, we used an in vitro assay to determine the amount of virus delivered by mosquitoes in their saliva. We found that Ae. aegypti as well as Ae. albopictus ensured a high replication of the virus which underwent an efficient dissemination as detectable in the salivary glands at day 2 post-infection (pi). Infectious CHIKV particles were delivered by salivary glands from day 2 with a maximum at day 6 pi for Ae. albopictus (10^3.3 PFU) and day 7 pi for Ae. aegypti (10^2.5 PFU).

Conclusions

Ae. albopictus is slightly more efficient than Ae. aegypti to transmit the variant E1-226V of CHIKV. These results will help to design an efficient vector control to limit transmission as soon as the first human cases are diagnosed.     

Population structure and genetic diversity of Aedes aegypti and Aedes albopictus in Penang as revealed by mitochondrial DNA cytochrome oxidase I

The population genetics study is crucial as it helps in understanding the epidemiological aspects of dengue and help improving a vector control measures. This research aims to investigate the population genetics structure of two common species of Aedes mosquitoes in Penang; Aedes aegypti and Aedes albopictus using Cytochrome Oxidase I (COI) mitochondrial DNA (mtDNA) marker. Molecular investigations were derived from 440 bp and 418 bp mtDNA COI on 125 and 334 larvae of Aedes aegypti and Aedes albopictus respectively, from 32 locations in Penang. All samples were employed in the BLASTn for species identification. The haplotype diversity, nucleotide diversity, neutrality test and mismatch distribution analysis were conducted in DnaSP version 5.10.1. AMOVA analysis was conducted in ARLEQUIN version 3.5 and the phylogenetic reconstructions based on maximum likelihood (ML) and neighbor-joining (NJ) methods were implemented in MEGA X. The relationships among haplotypes were further tested by creating a minimum spanning tree using Network version 4.6.1. All samples were genetically identified and clustered into six distinct species. Among the species, Ae. albopictus was the most abundant (67.2%), followed by Ae. aegypti (25.2%) and the rest were counted for Culex sp. and Toxorhynchites sp. Both Ae. aegypti and Ae. albopictus show low nucleotide diversity (π) and high haplotype diversity (h), while the neutrality test shows a negative value in most of the population for both species. There are a total of 39 and 64 haplotypes recorded for Ae. aegypti and Ae. albopictus respectively. AMOVA analysis revealed that most of the variation occurred within population for both species. Mismatch distribution analysis showed bimodal characteristic of population differentiation for Ae. aegypti but Ae. albopictus showed unimodal characteristics of population differentiation. Genetic distance based on Tamura-Nei parameter showed low genetic divergent within population and high genetic divergent among population for both species. The maximum likelihood tree showed no obvious pattern of population genetic structure for both Ae. aegypti and Ae. albopictus from Penang and a moderate to high bootstrap values has supported this conclusion. The minimum spanning network for Ae. aegypti and Ae. albopictus showed five and three dominant haplotypes respectively, which indicates a mixture of haplotypes from the regions analysed. This study revealed that there is no population genetic structure exhibited by both Ae. aegypti and Ae. albopictus in Penang. Mutation has occurred rapidly in both species and this will be challenging in controlling the populations. However, further analysis needed to confirm this statement.     

Chromosomal differentiation in two species of Aedes and their hybrids revealed by giemsa C-banding

Giemsa C-banding patterns in two species of mosquitoes, Aedes aegypti and Aedes mascarensis, their hybrids and backcross progeny revealed differences in the sex chromosome pair. In A. aegypti, the female determining or the m chromosome in both males and females shows a conspicuous band in the centromere region and another band in one arm. The male determining or the M chromosome is devoid of any bands. Progeny of crosses involving A. aegypti females and A. mascarensis males showed interesting albeit unexpected results. The intercalary band was suppressed in both sons and daughters. When such F₁ sons were backcrossed to A. aegypti females, a proportion of males developed into intersexes. These intersex progeny differed from the normal males in terms of their banding pattern. In the reciprocal cross (A. mascarensis female × A. aegypti male), the F₁ and the backcross progeny yielded the expected C-banding patterns. The implications of the reversible expression of the intercalary band on the A. aegypti m chromosome and its relevance to genetic regulation are discussed.     

Efficacy of targeted indoor residual spraying with the pyrrole insecticide chlorfenapyr against pyrethroid-resistant Aedes aegypti

BackgroundThere is an increased need to mitigate the emergence of insecticide resistance and incorporate new formulations and modes of application to control the urban vector Aedes aegypti. Most research and development of insecticide formulations for the control of Ae. aegypti has focused on their peridomestic use as truck-mounted ULV-sprays or thermal fogs despite the widespread knowledge that most resting Ae. aegypti are found indoors. A recent modification of indoor residual spraying (IRS), termed targeted IRS (TIRS) works by restricting applications to 1.5 m down to the floor and on key Ae. aegypti resting sites (under furniture). TIRS also opens the possibility of evaluating novel residual insecticide formulations currently being developed for malaria IRS.MethodsWe evaluated the residual efficacy of chlorfenapyr, formulated as Sylando 240SC, for 12 months on free-flying field-derived pyrethroid-resistant Ae. aegypti using a novel experimental house design in Merida, Mexico. On a monthly basis, 600 female Ae. aegypti were released into the houses and left indoors with access to sugar solution for 24 hours. After the exposure period, dead and alive mosquitoes were counted in houses treated with chlorfenapyr as well as untreated control houses to calculate 24-h mortality. An evaluation for these exposed cohorts of surviving mosquitoes was extended up to seven days under laboratory conditions to quantify “delayed mortality”.ResultsMean acute (24-h) mortality of pyrethroid-resistant Ae. aegypti ranged 80–97% over 5 months, dropping below 30% after 7 months post-TIRS. If delayed mortality was considered (quantifying mosquito mortality up to 7 days after exposure), residual efficacy was above 90% for up to 7 months post-TIRS application. Generalized Additive Mixed Models quantified a residual efficacy of chlorfenapyr of 225 days (ca. 7.5 months).ConclusionsChlorfenapyr represents a new option for TIRS control of Ae. aegypti in urban areas, providing a highly-effective time of protection against indoor Ae. aegypti females of up to 7 months.     

Bioassay of mosquito iridescent virus of Aedes taeniorhynchus in cell cultures of Aedes aegypti.

A bioassay of mosquito iridescent virus (MIV) of Aedes taeniorhynchus was developed using cell cultures of Aedes aegypti. The dilution end point technique was based on the occurrence of cytopathic effects which were optimum at 31°C. Peleg's A. aegypti cell line was more sensitive and reliable than Singh's A. aegypti cell line for infectivity titration of the “R” and “T” strains of MIV. The highest tissue culture infectivity dose 50s (TCID₅₀) were elicited by virion:cell ratios of approximately 10. TCID₅₀ titers were significantly reduced by virus neutralization with either homologous or heterologous antiserum to either RMIV or TMIV. The virus propagated in either cell line was not infectious to A. taeniorhynchus larvae, or to the respective cells from which the virus was produced. All plaque assay attempts were unsuccessful.     

Temperature and photoperiod effects on dormancy status and life cycle parameters in Aedes albopictus and Aedes aegypti from subtropical Argentina

Aedes albopictus (Diptera: Culicidae) distribution is bounded to a subtropical area in Argentina, while Aedes aegypti (Diptera: Culicidae) covers both temperate and subtropical regions. We assessed thermal and photoperiod conditions on dormancy status, development time and mortality for these species from subtropical Argentina. Short days (8 light : 16 dark) significantly increased larval development time for both species, an effect previously linked to diapause incidence. Aedes albopictus showed higher mortality than Ae. aegypti at 16 °C under long day treatments (16 light : 8 dark), which could indicate a lower tolerance to a sudden temperature decrease during the summer season. Aedes albopictus showed a slightly higher percentage of dormant eggs from females exposed to a short day, relative to previous research in Brazilian populations. Since we employed more hours of darkness, this could suggest a relationship between day‐length and dormancy intensity. Interestingly, local Ae. aegypti presented dormancy similar to Ae. albopictus, in accordance with temperate populations. The minimum dormancy in Ae. albopictus would not be sufficient to extend its bounded distribution. We believe that these findings represent a novel contribution to current knowledge about the ecophysiology of Ae. albopictus and Ae. aegypti, two species with great epidemiological relevance in this subtropical region.