Anti-mosquito antibodies decrease the reproductive capacity of Aedes aegypti

Aedes aegypti (L.) fed on rabbits immunized with mosquito antigens showed a reduction in fecundity in the first oviposition and decreased viability of the progeny. Feeding behaviour of mosquitoes was not affected and no significant mortality was observed due to the presence of anti-mosquito antibodies in the bloodmeal. Antibodies were detected in the oocytes of mosquitoes 48 h after the bloodmeal. The role of specific antibodies in influencing fecundity is discussed.     

Anti-mosquito antibodies and their effects on feeding, fecundity and mortality of Aedes aegypti

Anti-mosquito antibodies, produced in mice inoculated with mosquito homogenates or exposed to mosquito bites, reacted with a variety of mosquito antigens including muscle, gut, fat body and nervous tissue; those in anti-mosquito bite sera reacted solely with salivary glands. Mosquitoes fed on restrained immunized mice showed a significant increase in mortality correlated to both the titre and specificity of the anti-mosquito antibodies ingested. No changes in their fecundity or feeding success were noted. Mosquitoes exposed to unrestrained immunized mice or mosquito-bitten mice, however, showed a significant reduction in feeding success, possibly reflecting enhanced host grooming.     

白纹伊蚊和埃及伊蚊对基孔肯雅病毒的易感性和传播性的研究

用４株基孔肯雅病毒经口感染白纹伊蚊和埃及伊蚊,进行了易感性和传播性的研究。结果表明,这两种蚊虫对基孔肯雅病毒易感。无论白纹伊蚊或埃及伊蚊,感染后第５－６天即可通过吸血将病毒传播给乳鼠,至第８－１３天,传播率可高达５５．５５％－１００％。感染蚊亦可经叮咬将病毒传播给小鸡。埃及伊蚊的易感性和传播率高于白纹伊蚊。实验还发现,不同来源毒株之间存在一定差异,如分离自云南白纹伊蚊的Ｍ８１株的感染率和传播率均高于其它毒株。这些结果表明,白纹伊蚊和埃及伊蚊在基孔肯雅病毒的保存和传播中起重要作用。     

Insecticide susceptibility status of Aedes aegypti (Diptera: Culicidae) from Townsville

Insecticide susceptibility of a 1995 Townsville Aedes aegypti (L.) population was investigated in the laboratory according to World Health Organization guidelines. Using baseline data from two Townsville populations (1955 and 1989), larval bioassays detected significant increases in susceptibility to synthetic pyrethroids and malathion, and significant reductions in susceptibility to most organophosphates and propoxur. Adult bioassays detected significant resistance to bendiocarb and DDT. Comparison of larval data with the international reference ROCK strain showed substantial resistance to have developed to malathion and fenthion. Further analysis revealed the presence of distinct substrains in the baseline 1989 population, which displayed varying levels of temephos susceptibility. We concluded that susceptibility investigations should assess mosquito populations collected from many sites within an area rather than taking a single population from one site, and that the 1989 Ae. aegypti colony would be unsuitable for use as a susceptible reference population.     

Medical entomology: changes in the spectrum of mosquito-borne disease in Australia and other vector threats and risks, 1972–2004

Abstract   Our paper presents an assessment of research and operational development in relation to medically important mosquito-borne disease, mainly the arboviruses Ross River, Barmah Forest, Murray Valley encephalitis, Japanese encephalitis, Kunjin and dengue, but also with respect to malaria. Since 1972, there have been considerable gains in research output, organisational structure, communication, surveillance including quarantine inspection and operational control. This has been due to the 1974 epidemic of Murray Valley encephalitis virus extending into temperate Australia, increasing occurrence of Ross River and the dengue viruses, the discovery of Barmah Forest virus as a disease entity in 1988, and the introduction of Japanese encephalitis in 1995. Because many of the outputs involve methodologies of global import, this has resulted in an unprecedented upsurge in publications of international standard.     

Laboratory testing of a lethal ovitrap for Aedes aegypti

Laboratory tests were conducted to determine the feasibility of making the mosquito ovitrap lethal to Aedes aegypti (L.) when they attempt to oviposit in the trap. Heavy-weight velour paper strips (2.54 x 11 cm) were used as an alternative to the wooden paddle normally provided as a substrate for mosquito oviposition. The paper strips were pretreated with insecticide solutions and allowed to dry before being used in oviposition cups of 473 ml capacity, filled with water initially to within 2.5 cm of the brim. Insecticides chosen for their quick knock-down efficacy were bendiocarb 76% WP (1.06 mg a.i./strip) and four pyrethroids: permethrin 25% WP (0.16 mg a.i./strip), deltamethin 4.75% SC (0.87 mg a.i./strip), cypermethrin 40% WP (2.81 mg a.i./strip), and cyfluthrin 20% WP (0.57 mg a.i./ strip). For experimental evaluation, two oviposition cups (one with an insecticide-treated strip and one with an untreated strip) were placed in cages (cubic 30 cm) with gravid female Ae. aegypti mosquitoes (aged 6-8 days) from a susceptible laboratory strain. Mortality-rates of female mosquitoes were 45% for bendiocarb, 47% for permethrin, 98% for deltamethrin, 100% for cypermethrin, and 100% for cyfluthrin. Young instar larvae added to the treated cups died within 2h. After water evaporation from the cups for 38 days, fresh mosquito females had access to previously submerged portions of the velour paper paddle, and mortality rates of 59% or more occurred. Cups that had water (360 ml) dripped into them, to simulate rain, produced female mosquito mortality rates of > 50% and all larvae died within 3 h of being added. These tests demonstrate that the ovitrap can be made lethal to both adults and larvae by insecticidal treatment of the ovistrip. Field efficacy trials are underway in Brazil to access the impact of this simple, low-cost, environmentally benign approach on populations of the dengue vector Ae. aegypti.     

Lessons of Aedes aegypti control in Thailand

Abstract. The incidence of dengue haemorrhagic fever (DHF) in Thailand has increased cyclically since the first recognized outbreak in 1958. Without an effective vaccine against dengue, and considering the clinical difficulty of treating DHF cases, vector control is needed to prevent dengue transmission. Since the establishment of the WHO Aedes Research Unit in 1964, continued since 1973 as the WHO Collaborating Centre at the Department of Medical Research in Bangkok, much operational research has been carried out in Thailand on the bionomics and control of dengue vectors: Aedes aegypti and Ae.albopictus. This review shows that, as in most other countries, dengue vector control programmes in Thailand make little use of the procedures arising from research, nor have they reduced the upward trend of dengue or prevented DHF outbreaks. Implications of the reluctance to use results of operational research on vector control are considered and remedial suggestions made.     

The vector competence of Culex annulirostris, Aedes sagax and Aedes alboannulatus for Murray Valley encephalitis virus at different temperatures

Culex annulirostris Skuse, colonized from Brisbane, Queensland, and Mildura, Victoria, Australia, were effective vectors of Murray Valley encephalitis virus at 20, 27 and 32-35 degrees C with full extrinsic incubation periods of 15, 10 and 4 days respectively. At 20 degrees C, 7-11 days post-infection, transmission by the Mildura colony (0-20%) was less efficient than the Brisbane colony (30-70%) but both were capable of 75-100% transmission after longer extrinsic incubation periods. Discriminant analysis of body and salivary gland titres showed that these were not satisfactory indicators of transmission. Wild-caught Aedes sagax (Skuse) and Cx annulirostris from the Murray Valley showed equal competence, but Aedes alboannulatus (Macquart) was a poor vector. The results provide data on rural amplification of Murray Valley encephalitis virus during spring and suggest that further work on the potential of Ae. sagax as a natural vector is warranted.     

Population structure and dispersal of the saltmarsh mosquito Aedes vigilax in Queensland, Australia

Population genetics of the mosquito Aedes vigilax (Skuse) (Diptera: Culicidae), a major vector of arboviruses (e.g. Barmah Forest, Ross River), were investigated to obtain an indirect estimate of mosquito dispersal characteristics in typical habitats of Aedes vigilax in south-east Queensland: on the off-shore islands of Moreton Bay and on the mainland where disjunct breeding populations of Ae. vigilax are distributed along intertidal marsh. Six allozyme loci were assessed for genetic differentiation between samples from 11 localities. Significant larval variation between some breeding sites was attributed to site-specific selection. Nonsignificant genetic differentiation was found among collections of adult mosquitoes caught in light traps throughout the study area (exceeding 60x27 km), indicating widespread dispersal. As distances of < or = 9 km over water did not appear to act as effective barriers to Ae. vigilax dispersal, localized control activities applied to Ae. vigilax breeding sites are unlikely to be effective against the vagile adult population. Therefore, the contiguous shires programme of broad acre control is endorsed to prevent the spread of arboviruses carried by Ae. vigilax     

Proteome of Aedes aegypti in response to infection and coinfection with microsporidian parasites

Hosts are frequently infected with more than one parasite or pathogen at any one time, but little is known as to how they respond to multiple immune challenges compared to those involving single infections. We investigated the proteome of Aedes aegypti larvae following infection with either Edhazardia aedis or Vavraia culicis, and coinfections involving both. They are both obligate intracellular parasites belonging to the phylum microsporidia and infect natural populations of Ae. aegypti. The results found some proteins only showing modified abundance in response to infections involving E. aedis, while others were only differentially abundant when infections involved V. culicis. Some proteins only responded with modified abundance to the coinfection condition, while others were differentially abundant in response to all three types of infection. As time since infection increased, the response to each of the single parasite infections diverged, while the response to the E. aedis and coinfection treatments converged. Some of the proteins differentially abundant in response to infection were identified. They included two vacuolar ATPases, proteins known to have a role in determining the infection success of intracellular parasites. This result suggests microsporidia could influence the infection success of other intracellular pathogens infecting vector species of mosquito, including viruses, Plasmodium and Wolbachia.     

Modulation of Host Learning in Aedes aegypti Mosquitoes

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Discrepancies between Aedes aegypti identification in the field and in the laboratory after collection with a sticky trap

Currently, sticky traps are regularly employed to assist in the surveillanceof Aedes aegypti infestation. We tested two alternativeprocedures for specimen identification performed by local health agents: directly inthe field, as recommended by certain manufacturers, or after transportation to thelaboratory. A total of 384 sticky traps (MosquiTRAP) were monitored monthly duringone year in four geographically representative Brazilian municipalities. When thesame samples were inspected in the field and in the laboratory, large differenceswere noted in the total number of mosquitoes recorded and in the number of specimensidentified as Ae. aegypti by both procedures. Although fieldidentification has the potential to speed vector surveillance, these results point touncertainties in the evaluated protocol.     

Physiological and biochemical response of Aedes aegypti tolerance to Bacillus thuringiensis

Bacillus thuringiensis subsp. israelensis (Bti) represents the only eco-friendly bio-degradable insecticide for mosquito-borne disease control. Our research aims to identify if mosquito detoxification enzymes play an important role in Bti tolerance mechanisms in the dengue vector, Aedes aegypti. Several enzymes, such as amylase, cytochromes P450, Na⁺/K⁺-ATPase, acetylcholinesterase, protease and glutathione S-transferase (GST) were analysed and level of activity determined in Ae. aegypti larvae after Bti treatment. Bti exposure significantly increased the level of amylase (183.2%) as well as the activity of cytochromes P450 (177.5%), and Na⁺/K⁺-ATPase (142.9%). On the other hand, there was a decrease of 8.6% and 11.4% in acetylcholinesterase and GST activity, and no significant effect in the total level of protease activity. We suggest that the variation in amylase, cytochromes P450, Na⁺/K⁺-ATPase, acetylcholinesterase, protease and GST activity may be associated with the Bti insecticidal mechanism. This study provides the basis of detoxifying enzymes in Bti tolerance.     

Dissemination barriers to Ross River virus in Aedes vigilax and the effects of larval nutrition on their expression

Effects of larval nutrition on vector competence of the mosquito Aedes vigilax (Skuse) (Diptera: Culicidae) from Townsville, north Queensland, for Ross River virus (RR) were examined. Larvae were reared on three different diets to create three significantly different size classes of adult mosquito. These were fed on serial dilutions of RR and then sampled on alternate days so the progression of the virus through the mosquito could be examined. No differences of vector competence could be attributed to larval nutrition. Barriers to infection and the correlation between infection rate, viral titre and transmission of RR by Ae. vigilax were also examined. The mesenteronal barrier was the only infection barrier expressed. No correlation between viral titre and transmission was detected, but a strong correlation was found between salivary gland infection and transmission rate. From this it was possible to estimate that 98.7+/-1.3% of Ae. vigilax with infected salivary glands transmit RR.     

Cloning and characterization of a chitin synthase cDNA from the mosquito Aedes aegypti

Characterization of the enzymes involved in the chitin biosynthetic pathway in mosquitoes is critical due to the importance of chitin in the formation of the peritrophic matrix [PM] and its potential impact on vector competence. Chitin is the homopolymer of the amino sugar N-acetyl-D glucosamine [GlcNAc]. The final step of incorporation of GlcNAc into the chitin polymer is catalyzed by the enzyme chitin synthase [CS]. CS is a membrane bound enzyme, but the mechanism of its action in the biosynthesis of the PM is not understood. We have isolated and sequenced a CS-encoding cDNA clone from the mosquito Aedes aegypti, compared its sequence with CS from other organisms and studied its RNA expression. The cDNA is 3.5 kb in length with an open reading frame of 2.6 kb that encodes a protein of 865 amino acids with a predicted molecular mass of 99.5 kDa. The putative translation product shares 90% similarity to two CS proteins from Caenorhabditis elegans and 50% similarity to Saccharomyces cerevisiae in the catalytic domain of CS enzymes. Data suggest that CS is a single copy gene. RT-PCR analysis shows CS message in whole non-blood-fed females, whole blood-fed females, non-blood-fed midguts and in midguts dissected at different time points post-blood-feeding. In situ hybridization studies of midgut samples revealed that CS mRNA increases following a bloodmeal and is localized to the periphery of the epithelial cells facing the midgut lumen.     

Oral susceptibility of Aedes albopictus to dengue type 2 virus: a study of infection kinetics, using the polymerase chain reaction for viral detection

Female Aedes albopictus mosquitoes, aged 1 week, were infected with DEN-2 dengue virus. The kinetics of infection in mosquito brain and mesenteron were monitored using DNA probes with polymerase chain reaction (PCR) amplification of target DNA sequences coding for DEN-2 virus envelope protein, compared with the standard immunofluorescence assay technique (IFA). Rates of virus detection in the mesenteron of orally infected mosquitoes rose to 38% by day 4 post-inoculation, then declined until day 8, followed by irregular peaks around days 11-14 and subsequently. In mosquito head squashes, virus was detected from day 4 onwards, reaching 38% positive by day 18. Salivary glands of all the same females were found to be positive for virus by day 8 onwards. Parenterally infected Ae.albopictus females were all positive for DEN-2 in the brain and salivary glands 8 days post-inoculation. In every case, results obtained with the PCR matched those from the IFA. Our DNA probe with PCR procedure can therefore be utilized as a sensitive and reliable method for studies of DEN-2 vectors.     

Polymorphic microsatellite markers for studies of Aedes aegypti (Diptera: Culicidae), the vector of dengue and yellow fever

A significant challenge to population genetic studies of the dengue vector, Aedes aegypti, has been the lack of polymorphic microsatellite loci. In an effort to develop useful markers, we evaluated the genetic variation at 17 microsatellite loci identified in the A. aegypti genome. Nine loci with at least five alleles were identified in field‐collected specimens from Thailand. An additional two loci carried five alleles if samples from an A. aegypti laboratory colony were included. Our results greatly increase the number of highly variable markers available for the study of the genetics and the population structure of this medically important species.