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[Individual protection against insect vectors].

Many diseases for which no vaccine is available are transmitted by insect and arthropod vectors, the main exceptions being yellow fever and Japanese encephalitis B. Treatment is less and less effective due to the development of chemoresistance to therapeutic and prophylactic drugs as is well-illustrated by malaria. One of the best methods of preventing these diseases is personal protection against insect bites. Personal protection measures can be divided into three categories which can be used separately or in combination : application of repellents to the skin, wearing clothes impregnated with insecticides, and use of bed nets and other barriers impregnated with insecticides. The choice of method depends on the type of insect vector involved. For insects that are active during the day or at dusk, application of repellents to the skin gives good short-term protection and wearing impregnated clothes is useful. Bed nets that have been properly impregnated with pyrethroids are highly effective for night-time protection. Since personal protection methods are not 100% effective, they must be used in association with chemoprophylaxis according to medical guidelines. Medical advice should be sought if fever should occur especially after returning from a trip in the tropics.

Animals↗

Targeted deletion of the gp72 gene decreases the infectivity of Trypanosoma cruzi for mice and insect vectors.

The infective behavior of a mutant Trypanosoma cruzi clone, carrying a targeted deletion of the gp72 gene, was studied in the insect vector Triatoma infestans and in mice. After feeding T. infestans with complement-resistant forms (CRF) of Ynull and wild-type clones, it was observed that the number of parasites released in the bug's feces was reduced to less than 1% in the mutant clone. Both gp72-null and wild-type clones had a low infectivity for mice in comparison with other T. cruzi isolates, probably as a consequence of prolonged in vitro culture. Therefore, the behavior of both clones was tested in highly susceptible BALB suckling mice and immunodeficient athymic mice. After infecting the animals with 10(5) CRF, wild-type parasites could be detected in fresh blood mounts of most mice, but mutants were never found by this method. However, in 4 of 22 hemocultures from 11 athymic mice, gp72-null epimastigotes carrying the mutant phenotype were reisolated by day 29 of infection. Serological and polymerase chain reaction determinations performed on the blood of animals inoculated with the mutants indicated the possibility of temporary infections, which were extinguished after 90 days. The intact GP72 gene seems essential for sustaining latent infections in immunocompetent animals.

Animals↗

Resistance to insecticides in insect vectors of disease: est alpha 3, a novel amplified esterase associated with amplified est beta 1 from insecticide resistant strains of the mosquito Culex quinquesfasciatus.

Vector control programmes in many countries face the dual problems of parasite drug resistance and insecticide resistance in the insect vectors of the disease. Here we report for the first time a new esterase-based insecticide resistance mechanism in the filariasis vector Culex quinquefasciatus. The field collected COL strain of C. quinquefasciatus from Columbia was heterogeneous for organophosphorus insecticide resistance. On native polyacrylamide gels it had an elevated beta-naphthyl acetate specific esterase with the same Rf as that for the Est beta 1s involved in insecticide resistance in other strains of this mosquito species. After five generations of temephos insecticide selection, both the esterase specific activity with p-nitrophenyl acetate and the temephos LC50 values were increased, suggesting that elevation of esterase activity was the underlying mechanism of resistance. Western blots with antisera raised to Est alpha 2(1) and Est beta 2(1) from C. quinquefasciatus indicated that the COL strain had an elevated Est alpha 3 enzyme which co-migrated on native gels with Est beta 1. Southern blots indicated that an est alpha 3 gene was amplified in the COL strain and a Cuban mosquito strain (MRes), although the restriction digest patterns of the est beta 1 genes in these two strains are different. In contrast, the Californian TEMR strain, with the amplified est beta 1(1) gene, had no associated elevated Est alpha. Restriction digest patterns for COL and TEMR DNA suggest that they contain an identical est beta 1(1) gene, but our data suggest that the est alpha 3 gene occurs on the same amplicon as an est beta 1 gene although the genes are probably > 10 kb apart. Hence, either the COL strain has two est beta 1 genes or the est beta 1(1) amplicon in TEMR has been disrupted at some stage during the long colonisation of this strain and the amplified est alpha has been lost.

Animals↗

A sialidase activity in the midgut of the insect Triatoma infestans is responsible for the low levels of sialic acid in Trypanosoma cruzi growing in the insect vector.

Trypanosoma cruzi expresses a unique trans-sialidase that is responsible for the transfer of sialic acid from host glycoproteins and glycolipids to mucin-like glycoprotein acceptors on the parasite surface. The enzyme and the sialic acid acceptors are present in the mammalian forms of the parasite and in the parasite forms that grow in axenic cultures, which correspond to the developmental stages found in the insect vectors. Here we show that parasite forms growing in the vector Triatoma infestans express trans-sialidase in the hindgut portions of the insect. However, the sialic acid acceptors are poorly sialylated due to the low concentration of sialic acid donors in the gut lumen of T.infestans, which feeds exclusively on blood that is rich in sialic acid donors. These low levels of sialic acid donors are due to a novel sialidase activity present mainly in the anterior midgut with high specificity for alpha-2,3-sialyllactose, but not for alpha-2,6-sialyllactose. The activity is present in starved insects or insects fed with culture medium, indicating that it did not originate from the blood meal. Enzyme activity does not decrease in insects fed with antibiotics, is present in the salivary glands, and the few bacteria isolated from the gut and faeces of T.infestans did not display sialidase activity, indicating that the enzyme is not derived from a commensal organism. This novel activity could have a nutritional role in the gut of haematophagous insects and indicates that acquisition of sialic acid is not required for parasite development in the gut of T.infestans.

Animals↗

Rhythmic expression of the cycle gene in a hematophagous insect vector.

BACKGROUND: A large number of organisms have internal circadian clocks that enable them to adapt to the cyclic changes of the external environment. In the model organism Drosophila melanogaster, feedback loops of transcription and translation are believed to be crucial for the maintenance of the central pacemaker. In this mechanism the cycle (or bmal1) gene, which is constitutively expressed, plays a critical role activating the expression of genes that will later inhibit their own activity, thereby closing the loop. Unlike Drosophila, the molecular clock of insect vectors is poorly understood, despite the importance of circadian behavior in the dynamic of disease transmission. RESULTS: Here we describe the sequence, genomic organization and circadian expression of cycle in the crepuscular/nocturnal hematophagous sandfly Lutzomyia longipalpis, the main vector of visceral leishmaniasis in the Americas. Deduced amino acid sequence revealed that sandfly cycle has a C-terminal transactivation domain highly conserved among eukaryotes but absent in D. melanogaster. Moreover, an alternative form of the transcript was also identified. Interestingly, while cycle expression in Drosophila and other Diptera is constitutive, in sandflies it is rhythmic in males and female heads but constitutive in the female body. Blood-feeding, which causes down-regulation of period and timeless in this species, does not affect cycle expression. CONCLUSION: Sequence and expression analysis of cycle in L. longipalpis show interesting differences compared to Drosophila suggesting that hematophagous vector species might present interesting new models to study the molecular control of insect circadian clocks.

Amino Acid Sequence↗

Antiserum against perimicrovillar membranes and midgut tissue reduces the development of Trypanosoma cruzi in the insect vector, Rhodnius prolixus.

Antiserum raised against Rhodnius prolixus perimicrovillar membranes (PMM) and midgut tissue interfered with the midgut structural organization and reduced the development of Trypanosoma cruzi in the R. prolixus insect vector. SDS-PAGE and Western blot analyses confirmed the specific recognition of midgut proteins by the antibody. Feeding, mortality, molt, and oviposition of the insects were unaffected by feeding with the antiserum. However, the eclosion of the eggs were reduced from R. prolixus females treated with antiserum. Additionally, in vivo evaluation showed that after oral treatment with the antiserum, the intensity of infection with the Dm-28c clone of T. cruzi decreased in the digestive tract of fifth-instar nymphs and in the excretions of R. prolixus adults. These results suggest that the changes observed in the PMM organization in the posterior midgut of R. prolixus may not be important for triatomine survival but the antiserum acts as a transmission-reduction vaccine able to induce significant decreases in T. cruzi infection in the vector.

Animals↗

Management of plant viral diseases through chemical control of insect vectors.

In this article we consider the role of epidemiological factors and transmission processes of insect-vectored viruses on the effectiveness of insecticides in a disease management program. We also discuss the use of insecticides within the broader framework of the chemical environment surrounding vectors, and how chemical-induced alterations in the behavior of vectors can influence transmission. Our analysis confirms the belief of Heathcote, who stated in 1973, that "no one method of control is likely to keep crops entirely free from virus infection and as many preventative measures should be taken as are economically justified."

Journal Article↗

Induction of Trypanosoma cruzi metacyclogenesis in the gut of the hematophagous insect vector, Rhodnius prolixus, by hemoglobin and peptides carrying alpha D-globin sequences.

Trypanosoma cruzi, a protozoan responsible for the American trypanosomiasis (Chagas disease), multiplies and differentiates in the gut of triatomine insect vectors. The effects of hemoglobin and synthetic peptides carrying alpha D-globin fragments on both the growth and the transformation of T. cruzi epimastigotes (noninfective) into metacyclic trypmastigotes (infective forms) were studied. This differentiation in the insect's gut is expressed when hemoglobin and synthetic peptides corresponding to residues 30-49 and 35-73 of the alpha D-globin were added to the plasma diet. However, synthetic peptide 41-73 does not induce differentiation of epimastigotes even in the presence of the two former synthetic peptides. Thus, these data delineate an unusual molecular mechanism which modulates the dynamics of transformation of epimastigotes into metacyclic trypomastigotes in the triatomine vector's gut.

Amino Acid Sequence↗

Use of a real-time polymerase chain reaction-based fluorogenic 5' nuclease assay to evaluate insect vectors of Corynebacterium pseudotuberculosis infections in horses.

OBJECTIVE: To develop and use a sensitive molecular assay for detecting the phospholipase D (PLD) exotoxin gene of Corynebacterium pseudotuberculosis in an attempt to identify insect vectors that may be important in transmission of clinical disease in horses. SAMPLE POPULATION: 2,621 flies of various species. PROCEDURE: A real-time polymerase chain reaction (PCR)-based fluorogenic 5' nuclease (TaqMan) system (ie, TaqMan PCR assay) was developed for the detection of the PLD gene in insects. Flies were collected monthly (May to November 2002) from 5 farms in northern California where C. pseudotuberculosis infection in horses is endemic. Three of the 5 farms (which housed a total of 358 horses) had diseased horses during the study period. A total of 2,621 flies of various species were tested for the PLD gene of C. pseudotuberculosis. RESULTS: Evidence of bacterial DNA for the PLD gene was detected in skin biopsy specimens from clinically affected horses and from 3 fly species collected from farms where affected horses were housed. Farms with a high incidence of diseased horses had a high proportion of insects carrying the organism. High percentages of flies with positive results for the PLD gene were observed in October, when most clinically affected horses were observed. CONCLUSIONS AND CLINICAL RELEVANCE: Our results are consistent with the hypothesis that C. pseudotuberculosis may be vectored to horses by flies. Three potential vectors were identified, including Haematobia irritans, Stomoxys calcitrans, and Musca domestica. The organism can be identified in up to 20% of house flies (Musca domestica) in the vicinity of diseased horses.

Animals↗

Development and interactions of Trypanosoma cruzi within the insect vector.

Transmission of Chagas disease or American trypanosomiasis depends on Trypanosoma cruzi development and differentiation within its triatomine insect vector. In this review, Eloi Garcia and Patricia de Azambuja aim to outline the current areas of research that may explain aspects of the parasite-vector interaction.

Journal Article↗

Action of isometamidium chloride on the insect vector form of Trypanosoma vivax.

The effect of isometamidium chloride upon developing and mature Trypanosoma vivax occurring in Glossina palpalis palpalis flies was evaluated. Newly emerged G p palpalis flies were infected with T vivax by allowing them to feed on parasitaemic animals. Two experiments were conducted and in each the flies were divided into two groups. One group of infected flies was fed in vitro through a membrane on defibrinated cow blood containing isometamidium chloride at 0.1 mg ml-1, after which they were dissected and examined for trypanosomes, and the other group was fed in the same way on unmedicated blood. The results showed that out of a total number of 129 flies which fed on medicated blood, none was infected, while 55 out of 127 flies which fed on unmedicated blood were infected. The results indicate that isometamidium chloride eliminated the insect vector form of T vivax. These findings are of potential significance in the control of trypanosomiasis in the field, particularly in the operation of the sterile insect technique.

Animals↗

Electron microscopy of the formation of wound tumor virus in abdominally inoculated insect vectors.

Fifth-instar nymphs of Agallia constricta leafhoppers were injected abdominally with extracts from root tumors confining wound tumor virus (WTV). The insects were sacrificed at predetermined intervals, their internal organs dissected, fixed, embedded, sectioned, stained, and examined in a Siemens Elmiskop I. Sequential stages in virus development were reconstructed from consecutive samples of fatbody tissues. Changes resulting from the infection were: (i) a viroplasm, i.e., an accumulation of electron-dense aggregates; (ii) the appearance at the periphery of the viroplasm of a few fully formed virus particles recognized as virions; (iii) the formation of increasing numbers of individual virions, not only at the periphery but also in the viroplasm; (iv) the engulfing of virions within multimembranous structures; and (v) the formation of virus microcrystals either at the sites of former viroplasms, or at some distance. These morphological findings indicate that, following abdominal inoculation of WTV, the plant-pathogenic virus develops within the cytoplasmic matrix proper of insect vector cells. In addition to the fatbody tissues, WTV was detected in the epidermis, muscles, and trachea of abdominally inoculated insects, demonstrating the systemic invasion of the mechanically infected arthropod host. No virus was found in the gut tissues.

Journal Article↗