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PubMed2002

What's buzzing? Mosquito genomics and transgenic mosquitoes.

Genome projects and associated technologies are now being established for mosquito species that are vectors of human disease. The recent announcement of an award by the National Institute of Allergy and Infectious Diseases (NIAID) to Celera Genomics to sequence the Anopheles gambiae genome will further accelerate the completion of the sequencing of this genome. Completion of the An. gambiae sequence will mean that the genomes of all three organisms involved in the transmission of falciparum malaria--the mosquito, the parasite, and the human--will have been sequenced. This will greatly facilitate the identification of genes and pathways involved in the transmission of malaria. The recent genetic transformation of An. gambiae with the piggyBac transposable element and the transformation of another important malarial vector, Anopheles stephensi using the Minos element, now provide researchers with powerful tools with which to genetically manipulate these medically important vector species. Here we review the recent progress made in the extension of contemporary tools of modern genetics and genomics into these medically important insects.

Aedes↗
PubMed1984

Mosquito protein microassay. I. Protein determinations from small portions of single-mosquito homogenates.

A microassay is described for determining protein levels in individual mosquitoes. This method requires only a small portion of a homogenate of a single mosquito. Protein levels in Anopheles albimanus are positively correlated with body weight. Protein levels for different geographic strains of Anopheles albimanus and different generations of laboratory rearings are compared.

Animals↗
PubMed1998

Localization of the mosquito insulin receptor homolog (MIR) in reproducing yellow fever mosquitoes (Aedes aegypti).

The female mosquito takes a blood meal to produce a batch of eggs. Initiation of egg maturation and growth of oocytes is governed by several endocrine factors. Peptide factors from the brain are involved in this process and some are also responsible for the induction of ecdysone secretion. The latter appears to be required to maintain a high rate of vitellogenin synthesis. By analogy with the known functions of insulin-like molecules (e.g. bombyxins) which in insects activate the secretion of ecdysteroids, we have postulated that there is an insulin receptor homolog responsible for activation of endysone secretion in the ovary. We have recently cloned the mosquito homolog (MIR) and are now investigating its spatial and temporal distribution. Here, we have localized the insulin receptor (MIR) both at the mRNA and protein level using in situ-hybridization and immunocytochemistry. The receptor is expressed before a blood meal mainly in the nurse cells of ovaries. After a meal, follicle and nurse cells contain mRNA coding for the receptor. The intensity of expression rises in the follicle cells until they degenerate during choriogenesis. Immunocytochemical localization confirms the in situ data: the protein is present before and after a meal. Both methods confirm our previous findings by Northern blot analysis, in which the ovary was found to be the main source of the receptor mRNA. The dynamics of receptor mRNA are related to the dynamics of ecdysone secretion and its action on physiological processes.

Journal Article↗
PubMed1997

Microbial control of mosquitoes: management of the upper rhine mosquito population as a model programme.

For more than a decade, Bacillus thuringiensis israelensis (Bti) and B. sphaericus have been used successfully as biological control agents against mosquitoes in Germany. Over 1000 km(2) of breeding areas have been treated with Bti, resulting in a yearly reduction of the mosquito population by more than 90%, without evidence of any harmful impact on the environment. Here, Norbert Becker describes the control strategy and operational use of Bti and B. sphaericus in Germany.

Journal Article↗
PubMed2003

A plasmid encoding a combination of mosquito-larvicidal genes from Bacillus thuringiensis subsp. israelensis and Bacillus sphaericus confers toxicity against a broad range of mosquito larvae when expressed in Gram-negative bacteria.

A recombinant plasmid harboring cry4A, cry4B and cry11A from Bacillus thuringiensis subsp. israelensis and binary toxin genes from Bacillus sphaericus has been constructed. The three cry genes were placed under the control of the cry4B promoter whereas the binary toxin gene was controlled by its native promoter. The expression of toxins in Escherichia coli harboring the resulting plasmid, p4BDA-5142, was investigated. Cry4B expression was highest compared to other toxins. Although the level of toxin expression was low compared with E. coli expressing single toxins, the recombinant E. coli strain harboring p4BDA-5142 exhibited broad range mosquito-larvicidal activity against all Aedes, Culex and Anopheles larvae. This work has shown that the development of the recombinant plasmid can be used to broaden the host range spectrum of the appropriate bacterial host for mosquito control.

Aedes↗
PubMed1998

Similarities to a LINE element shared by Anopheline and Culicine mosquitos map to the distal end of dihydrofolate reductase amplicons in Aedes albopictus mosquito cells.

To extend our understanding of amplicon structure in methotrexate-resistant Mtx-5011-256 Aedes albopictus mosquito cells, we examined a series of cosmids containing genomic DNA corresponding to the unique 3'-end of the Type 1 dihydrofolate reductase amplicon. Cosmid pWED118 contained five EcoRI fragments ranging from 2 to 5 kb (A, B, C, F, G) that hybridized to cDNA from methotrexate-resistant cells. Of these, fragments B and F hybridized weakly to first-strand cDNA from sensitive cells and shared considerable sequence identity. Fragment G occurred twice in the map of pWED118; one copy mapped within a 10 kb BssHII core fragment from the Type I amplicon and a second copy mapped downstream in the 48 kb BssHII core fragment. Hybridization signals among fragments contained in overlapping cosmids suggested that a branch point defining two or more subtypes of the Type I amplicon occurs within or near the 10 kb BssHII genomic DNA fragment. A 1.8 kb sequence common to fragments B and F included an approximately 0.4 kb region that shared sequence similarities with a LINE element from Aedes aegypti and with a repeated sequence from Anopheles gambiae. In addition, these elements shared amino acid similarity to a reverse transcriptase from the nematode, Caenorhabditis elegans. Shared sequence between Aedes and Anopheles elements supports the hypothesis that an ancestral LINE-like element was active in mosquito genomes prior to the divergence of the subfamilies Culicinae and Anophelinae. The presence of homologies to LINE-like elements near a branch point in the dihydrofolate reductase amplicon is consistent with a possible role of repeated sequences in amplicon shortening.

Animals↗
PubMed1995

Immunoglobulins specific to mosquito salivary gland proteins in the sera of persons with common or hypersensitive reactions to mosquito bites.

Using the immunoblot technique, we analyzed the quality and quantity of IgG, IgG4, and IgE specific to mosquito salivary gland (hereafter abbreviate as SG) components of Aedes albopictus in the sera of volunteers with common reactions and of 3 patients with severe reactions. In the volunteers with delayed reactions only or with both delayed and immediate reactions, IgG against SG components of A. albopictus formed several faint or moderately stained bands. Those with immediate reactions showed several intense bands and many other weak bands. In volunteers, who had been bitten by Aedes sp. frequently but had no skin reaction, and in severe cases, many intense IgG bands were observed. IgG4 bound to SG components were found in the sera of the common reaction group at the levels of 24 and 48 kD, but, in one severe case, no bands were observed, although the total IgG was very high. IgE levels specific to SG components were much higher in severe cases than in the volunteers. These results indicate that high titers of specific IgG and IgE and lack of IgG4 for particular components of SG may lead to severe allergic reactions in severe cases. Immunoblotting analysis of the antibodies also verified the possibility of developing in vitro tests to identify causative species of the mosquito for severe cases.

Adult↗
PubMed1997

Analysis of LaCrosse virus S mRNA 5' termini in infected mosquito cells and Aedes triseriatus mosquitoes.

Nucleotide sequences were determined for the 5' termini of La Crosse virus (LAC) S segment mRNA from persistently infected mosquito cell cultures (C6/36 from Aedes albopictus) and embryos (Aedes triseriatus). LAC primes transcription of its mRNA with "scavenged" 5' caps and adjacent oligonucleotides from host mRNAs, and these non-virus-encoded 5'-terminal extensions are heterogeneous in infected mammalian cells. The nature of mosquito host-derived primers has not been previously investigated. During early C6/36 cell infection, LAC mRNA 5'-terminal sequences were heterogeneous, but variability decreased as infection persisted. One predominant sequence, 5' CCACTCGCCACT (sequence 1), was observed throughout C6/36 cell infection but was more prevalent after 15 days postinfection. This LAC mRNA 5'-terminal sequence comprised 81% of the scavenged host oligonucleotides from vertically infected A. triseriatus eggs during embryogenesis. As these embryos progressed in the dormant overwintering stage (diapause), the predominant scavenged sequence became 5' AGGAAAAGATGGT (sequence 2), and sequence 1 became less prevalent. As the eggs emerged from diapause, the LAC mRNA 5' termini were more variable; 33% had sequence 1, and the remainder were heterogeneous. In post-diapausing eggs, 100% of viral mRNAs had sequence 1 at their 5' termini. Molecular analyses thus revealed continuous but selective LAC cap scavenging during persistent C6/36 cell infection and during embryogenesis and diapause in A. triseriatus eggs. The variety of host-derived sequences was limited in both biosynthetically active (embryonating) and dormant (diapausing) eggs.

Aedes↗
PubMed2006

Mosquitoes of the mangrove forests of India: Part 3--Andaman and Nicobar Islands, including an update on the mosquito fauna of the islands.

ABSTRACT. Fifty-three mosquito species belonging to 20 subgenera and 18 genera--Aedes, Anopheles, Armigeres, Culex, Downsiomyia, Finlaya, Heizmannia, Kenknightia, Lorrainea, Lutzia, Ochlerotatus, Orthopodomyia, Rhinoskusea, Stegomyia, Toxorhynchites, Tripteroides, Uranotaenia, and Verrallina--were recorded in the mangroves of Andaman and Nicobar Islands, India, in collections made during May and June 2001. Larvae of 43 species were collected from different mangrove habitats. Together with collections made in nonmangrove areas, 83 species in 22 subgenera and 20 genera in total were recorded of which 33 species are new records for the Andaman and Nicobar Islands. Rhinoskusea wardi, Fl. flavipennis, and Ve. consonensis are 3 new country records for India. Collections included topotype specimens of St. seampi. Together with species known from earlier records, the mosquito fauna of Andaman and Nicobar Islands is updated to 107 species in 23 subgenera and 24 genera.

Aedes↗
PubMed2006

Vector competence of Peruvian mosquitoes (Diptera: Culicidae) for a subtype IIIC virus in the Venezuelan equine encephalomyelitis complex isolated from mosquitoes captured in Peru.

We evaluated mosquitoes collected in the Amazon Basin, near Iquitos, Peru, for their susceptibility to a subtype IIIC strain of the Venezuelan equine encephalomyelitis complex. This virus had been previously isolated from a pool of mixed Culex vomerifer and Cx. gnomatos captured near Iquitos, Peru, in 1997. After feeding on hamsters with viremias of about 10(8) plaque-forming units of virus per ml, Cx. gnomatos was the most efficient vector. Other species, such as Ochlerotatus fulvus and Psorophora cingulata, although highly susceptible to infection, were not efficient laboratory vectors of this virus due to a significant salivary gland barrier. The Cx. (Culex) species, consisting mostly of Cx. (Cux.) coronator, were nearly refractory to subtype IIIC virus and exhibited both midgut infection as well as salivary gland barriers. Additional studies on biting behavior, mosquito population densities, and vertebrate reservoir hosts of subtype IIIC virus are needed to determine the role that these species play in the maintenance and spread of this virus in the Amazon Basin region.

Animals↗
PubMedDate not supplied

[A comparative analysis of the potential epidemiological importance of mosquitoes in the genus Anopheles under the conditions of the anthropogenic landscapes in the Chu Valley, Kyrgyz Republic. 1. The activity of the attack of malarial mosquitoes on man].

Six species of the genus Anopheles (An. claviger were prevalent, An. messeae and An. hyrcanus were intermediate, and An. martinius, An. algeriensis and An. superpictus were rare) were recorded in the Chu [correction of Chuisk] Valley in the north of the Republic of Kirghizia. Due to the fact that the malaria situation was worsening, the potential epidemiological significance of Anopheles that predominate under the man-made districts of the region under study was comparatively analyzed. Mosquitoes were collected in the 1991-1992 seasons. This paper gives data on the seasonal attacking activity of the predominant Anopheles species in the foothills and lowlands of the Chu River valley. The maximum seasonal attacking activity of mosquitoes has been found to coincide with the time of their greatest numbers and to be in the hot summer period (July-August) that makes it favourable for the causative agents of malaria to develop.

Animals↗
PubMedDate not supplied

[A comparative analysis of the potential epidemiological importance of mosquitoes in the genus Anopheles under the conditions of the anthropogenic landscapes in the Chu Valley, Kyrgyz Republic. II. The spread of malarial mosquitoes by the types of day resting places].

The distribution of malaria mosquitoes was analyzed by the types of day's rest within the farmsteads to determine what feed different Anopheles species preferred in the Chu River valley, which is the most densely populated in the Republic of Kirghizia. Field trials were made in the April to October of 1991 and 1992. Above 12,000 mosquito imagoes were collected. The predominant An. claviger was found mainly in the living spaces of the farmsteads which were located in vicinity of breeding sites. This exophilic species readily attacks both cattle and man who have found themselves in the places of its day's rest. An. messeae was chiefly detected indoors. The species readily attacks human beings. In the valley of the Chu River, A. hyrcanus actively attacks man indoors. In the Chu River valley, An. martinius is a completely exophilic type which prefers the accumulation of large prey.

Animals↗
PubMed2005

Decrease of mosquito salivary gland proteins after a blood meal: an implication for pathogenesis of mosquito bite allergy.

Salivary gland protein profiles ofAedes aegypti (L.) and Culex quinquefasciatus (Say) pre- and post-blood feeding were analyzed. SDS-PAGE studies before blood feeding of Ae. aegypti demonstrated 8 major polypeptide bands of 20, 35, 37, 42, 45, 47, 70 kDa and a high molecular weight band >118 kDa, whereas those of Cx. quinquefasciatus demonstrated 9 major polypeptide bands of 20, 26, 36, 38, 45, 47, 49 kDa and 2 high molecular weight bands >118 kDa. After a blood feeding, salivary gland polypeptides of Ae. aegypti at 35, 37, 45, 47, 70 kDa and high molecular weight band >118 kDa were depleted, while the polypeptide bands of 20, 26, 36, 38 kDa were depleted in Cx. quinquefasciatus. The presented study suggests that these major polypeptides were introduced into vertebrate hosts when a mosquito took a blood meal. Further investigation in molecular, biochemical and immunological aspects of these salivary gland polypeptides may provide information for better understanding in the role of these proteins in mosquito bite allergy.

Aedes↗
PubMed1994

[The effect of Culex family mosquito larva on the sensitivity of Anopheles mosquitos with various karyotypes to the entomopathogenic bacteria Bacillus thuringiensis subsp. Israelensis].

Culex (C. modestus) and Anopheles (A. beklemishevi and A. messeae) larvae mosquito were treated with Bacillus thuringiensis subsp. israelensis (Bti) bacterium together and separately. It was determined that the presence of C. modestus reduced the mortality of Anopheles larvae and altered the results of the selection of genotypes resistant to Bti of the polymorphic species of A. messeae, C. modestus larvae consumed the pathogen more rapidly and protected the sensitive individuals of A. messeae with "northern" chromosomal inversions from destruction. The correlation of the selection of cohabiting Anopheles and Culex mosquitoes as well as the relation between the specific diversity and the stability of communities are discussed.

Animals↗
PubMed2006

Evaluation of synthetic repellents on mosquito nets in experimental huts against insecticide-resistant Anopheles gambiae and Culex quinquefasciatus mosquitoes.

Owing to the development of pyrethroid resistance in Anopheles gambiae, there is a need to develop chemical alternatives for use on mosquito nets. Synthetic insect repellents are widely used for personal protection as skin or clothing applications. The efficacy of repellent-treated nets (RTN) was evaluated in experimental huts in Côte d'Ivoire against pyrethroid-resistant populations of An. gambiae and Culex quinquefasciatus. The repellents tested were DEET (N,N-diethyl-3-methylbenzamide) at 7.9 g/m2 and two formulations of ethyl butylacetylaminopropionate (IR3535) at 7.6 g/m2 and 7.3 g/m2. Over 45 nights there was a 74-82% reduction in the number of An. gambiae entering the huts containing RTNs but no significant reduction in entry of C. quinquefasciatus. There was a 63-64% reduction in the proportion of An. gambiae blood feeding but no reduction in the proportion of C. quinquefasciatus blood feeding in huts with RTNs. An unexpected result was the 69-76% mortality of An. gambiae and 51-61% mortality of C. quinquefasciatus in huts containing RTNs. Treated filter paper bioassays in WHO test kits confirmed that confined contact with DEET induces mortality. The DEET-based product provided better and longer protection; tunnel test bioassays confirmed that residual activity lasted for up to 6 weeks. Application of repellents to nets warrants further investigation and development.

Animals↗
PubMed2005

Mosquito larvicidal activity of active constituent derived from Chamaecyparis obtusa leaves against 3 mosquito species.

Mosqutio larvicidal activity of Chamaecyparis obtusa leaf-derived materials against the 4th-stage larvae of Aedes aegypti (L.), Ochlerotatus togoi (Theobald), and Culex pipiens pallens (Coquillett) was examined in the laboratory. A crude methanol extract of C. obtusa leaves was found to be active (percent mortality rough) against the 3 species larvae; the hexane fraction of the methanol extract showed a strong larvicidal activity (100% mortality) at 100 ppm. The bioactive component in the C. obtusa leaf extract was characterized as beta-thujaplicin by spectroscopic analyses. The LC50 value of beta-thujaplicin was 2.91, 2.60, and 1.33 ppm against Ae. aegypti, Oc. togoi, and Cx. pipiens pallens larvae. This naturally occurring C. obtusa leaves-derived compound merits further study as a potential mosquito larval control agent or lead compound.

Aedes↗
PubMed1987

An economic assessment of the benefits of mosquito abatement in an organized mosquito control district.

A case study application of contingent valuation to the measurement of mosquito program benefits is presented. Annual program benefits in Jefferson County, TX are estimated to exceed costs by 1.8 times. Mean household benefits are $22.44 for owners and $18.96 for renters. Using ordinary least squares procedures these household benefits were found to be related to household socioeconomic characteristics, effectiveness of control efforts and environmental concerns.

Cost-Benefit Analysis↗
PubMed1970

Evaluation of compounds for insecticidal activity on adult mosquitos. 2. Toxicity to adult mosquitos and the physical properties of some new carbamates.

The N,N-dimethylcarbamates are much less toxic and much more volatile than the corresponding N-methylcarbamates. N-acylation of N-methylcarbamates is also generally accompanied by a loss in intrinsic toxicity to adult mosquitos by topical application, but the reduction in toxicity may be relatively small and in some N-acetyl derivatives may not occur at all. N-acylation is also accompanied by changes in physical properties such as lipid solubility and partition coefficient, which influence contact toxicity, and volatility, which affects residual activity. In the series of compounds examined, contact toxicity is maximal in the N-acetyl derivatives but their volatility is often so high that they lack the persistence required of residual insecticides. This does not apply, however, to the N-acetyl derivatives of OMS-597 and OMS-708 because the volatility of the parent compound is so low that some degree of persistence can be sacrificed to obtain some improvement in contact activity.OMS-1064 is the only N-chloroacetyl derivative tested so far that shows promise as a residual insecticide.

Animals↗