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[Basic trends in the research for the control of infected blood-sucking ticks and insects, the vectors of human diseases].

Attention was given to the necessity of control of the infected part of the population of vectors of transmissible infections. For this purpose it was suggested to investigate the nature of parasitism of the agent, to look for synergists of their pathogenic effect and to study the ecology of vectors in order to use control measures against the infected part of the population in the sites of aggregation.

Animals↗
PubMed1995

The future of microbial insecticides as vector control agents.

Insect vectors of human diseases are subject to diseases of their own caused by viruses, bacteria, fungi, protozoans, and nematodes. Over the past 30 years, many members of these groups have been evaluated as vector control agents, particularly for mosquito control. Most pathogens and nematodes occur primarily in larvae, and are only effective against this stage. The principal candidate control agents studied include iridescent and nuclear polyhedrosis viruses, the bacteria Bacillus thuringiensis and Bacillus sphaericus, the fungi Lagenidium giganteum, Culicinomyces clavosporus, and species of the genus Coelomomyces, the protozoan Nosema algerae, and the mermithid nematode Romanomermis culicivorax. Of these, the only one considered an operational success is the bacterium, Bacillus thuringiensis subsp. israelensis (B.t.i.), which has proven useful for control of both mosquito and blackfly larvae in programs where larviciding has been traditionally employed as a vector control tactic. The reasons for the success of B.t.i. are its cost-effectiveness and relative ease of use, which are due, respectively, to the ability of B.t.i. to be grown on artificial media and the development of formulations that can be applied using conventional insecticide application technology. Because few microbial insecticides are cost-effective, and those that are are only effective against larvae, these agents will likely play only a minor, but in some cases important, role in most future vector control programs.

Animals↗
PubMed1997

Long-term association of tomato yellow leaf curl virus with its whitefly vector Bemisia tabaci: effect on the insect transmission capacity, longevity and fecundity.

The association between tomato yellow leaf curl geminivirus (TYLCV, Israeli isolate) and its insect vector, the whitefly Bemisia tabaci, was investigated. Insects that emerged during a 24 h period were caged with TYLCV-infected plants for a 48 h acquisition access period, then with egg-plants--a TYLCV non-host--for the rest of their lives. While TYLCV DNA was associated with the whiteflies during their entire adult life, the amount of capsid protein rapidly decreased and was not detectable in the insect after approximately 12 days of age. The ability of the infected whiteflies to transmit TYLCV to tomato test plants steadily decreased with age but did not disappear completely. Transmission by viruliferous insects decreased from 100% to 10-20% during their adult lifetime, compared with a decrease from 100% to 50% for non-viruliferous insects. The association of TYLCV with adult B. tabaci led to a reduction of 17-23% in their life expectancy compared with insects that had not acquired the virus, and to a 40-50% decrease in the mean number of eggs laid. These results suggest that TYLCV has some features reminiscent of an insect pathogen.

Animals↗
PubMed1995

Immunological strategies for control of insect disease vectors: a critical assessment.

Insect-borne diseases are one of the major causes of disease and death in the tropical world. Conventional methods of disease control have proven insufficient and there is pressing need for devising new strategies. One approach that has been explored by several laboratories is to compromise vector fecundity and survivorship through the immunization of vertebrate hosts with the vector's internal organs (concealed antigens). Here, Marcelo Jacobs-Lorena and Francisco J.A. Lemos critically review the results obtained to date by use of this approach. It appears that the published work is less rigorous than would be desirable and the results are contradictory. In contrast to the successes obtained with a similar immunization strategy implemented for tick-borne diseases, it is not yet clear whether or not such an approach can be applied to insect-borne diseases.

Journal Article↗
PubMed2004

Tn5 as an insect gene vector.

The purpose of this study was to explore alternatives to insect-derived transposable elements as insect gene vectors with the intention of improving existing insect transgenesis methods. The mobility properties of the bacterial transposon, Tn5, were tested in mosquitoes using a transient transposable element mobility assay and by attempting to create transgenic insects. Tn5 synaptic complexes were assembled in vitro in the absence of Mg(2+) and co-injected with a target plasmid into developing yellow fever mosquito, Aedes aegypti, embryos. Target plasmids recovered from embryos a day later were screened for the presence of Tn5. Recombinants (transposition events) were found at a frequency of 1.2 x 10(-3). Some transposition events did not appear to be associated with canonical 9 bp direct duplications at the site of insertion and also were associated with either deletions or rearrangements. A Tn5 element containing the brain-specific transgene, 3 x P3DsRed, was assembled into synaptic complexes in vitro and injected into pre-blastoderm embryos of Ae. aegypti. Of the approximately 900 embryos surviving injection and developing into adults, two produced transgenic progeny. Both transgenic events involved the co-integrations of approximately five elements resulting in nested and tandem arrayed Tn5::3 x P3DsRed elements. This study extends the known host range of Tn5 to insects and makes available to insect biologists and others another eukaryotic genome-manipulation tool. The hyperactivity of synaptic complexes may be responsible for the unusual clustering of elements and managing this aspect of the element's behavior will be important in future applications of this technology to insects.

Aedes↗
PubMed1987

Expression of envelope glycoproteins of human immunodeficiency virus by an insect virus vector.

The envelope gene of human immunodeficiency virus was inserted into the genome of an insect virus vector (Autographa californica nuclear polyhedrosis virus). Upon infection of tissue culture cells, this recombinant virus produced immunoreactive polypeptides related to the envelope glycoproteins of human immunodeficiency virus. Serological survey indicates such polypeptides would be of value as antigens in diagnostics for acquired immunodeficiency syndrome.

Animals↗
PubMed1999

Recombinant baculoviruses as expression vectors for insect and mammalian cells.

Baculovirus expression vectors are widely used for expressing heterologous proteins in cultured insect cells. Recent advances include further development of the system for production of multi-subunit protein complexes, co-expression of protein-modifying enzymes to improve heterologous protein production, and additional applications of baculovirus display technology. The application of modified baculovirus vectors for gene expression in mammalian cells continues to expand.

Animals↗
PubMed1987

Use of the silkworm, Bombyx mori, and an insect baculovirus vector for high-level expression and secretion of biologically active mouse interleukin-3.

Using the virus vector derived from a baculovirus of Bombyx mori (Bm), we constructed an infectious recombinant virus carrying the mouse interleukin-3 (IL-3) cDNA placed downstream from the polyhedrin promoter. Silkworms infected in vivo with recombinant virus or the silkworm-derived BmN cell line infected in vitro secreted large amounts of IL-3 into hemolymph or culture medium, respectively. On a per volume basis, about 20-fold more activity was found in the culture supernatants of the infected BmN cells and 10000-fold more activity was detected in the hemolymph as compared to supernatants obtained from COS7 monkey cells transfected with plasmid pcD-IL3 using the SV40 early promoter [Yokota et al., Proc. Natl. Acad. Sci. USA 81 (1984) 1070-1074]. Three distinct species of Il-3 of molecular masses, 18, 20 and 22 kDa were produced and all were converted to a 15-kDa protein by N-glycanase digestion, indicating that silkworm cells glycosylated IL-3. The N-terminal amino acid sequences of the IL-3 purified from tissue culture medium and hemolymph were identical to that of mammalian-derived IL-3, showing that silkworm cells recognized the mammalian signal sequence and cleaved it at the correct position. The purified silkworm-produced IL-3 had biological activities indistinguishable from IL-3 produced by mammalian cells as assessed by mast-cell proliferation assays, colony-formation assays using mouse bone marrow cells, and by receptor-binding assays using [125I]IL-3.

Animals↗
PubMed2002

Detection of rice grassy stunt tenuivirus nonstructural proteins p2, p5 and p6 from infected rice plants and from viruliferous brown planthoppers.

The genome of Rice grassy stunt virus (RGSV) consists of 6 ambisense RNA segments, among which RNAs 1, 2, 5 and 6 are equivalent to RNAs 1, 2, 3 and 4, respectively, of Rice stripe virus, the type species of the genus Tenuivirus. The RGSV 36-kDa nucleocapsid protein (N) is encoded on the complementary strand of RNA 5. Here, we studied accumulation of three nonstructural proteins, a 23-kDa p2 protein encoded on vRNA 2 (virus genomic strand), a 22-kDa p5 protein encoded on vRNA 5, and a 21-kDa p6 protein encoded on vRNA 6, from RGSV-infected rice leaf tissues and from viruliferous vector insects (brown planthopper, Nilaparvata lugens) by Western blot analyses. p2, p5 and p6 were detected from RGSV-infected rice leaf homogenates; p2 was detected mostly in the cytoplasmic soluble fraction but also a small amount was detected in the cell-wall, organelle-enriched and crude membrane fractions; p5 and p6 were detected from the cytoplasmic soluble fraction in large amounts. Among individual nymphs of N. lugens raised on RGSV-infected rice leaves, only 20% of insects were positive with the N protein. A large amount of p5 was detected from all the N-positive insects. Small amounts of p2 and p6 were detected only from a subset of the N- and p5-positive insects. p5 may have an essential role in virus infection in both plant and insect hosts, whereas p2 may function in plants such as a cell-to-cell movement protein.

Animals↗
PubMed1991

Characterization of Trypanosoma cruzi from Argentina by electrophoretic zymograms.

Polyacrylamide gel electrophoretic patterns for six enzymes in 73 isolates and 38 clones of Trypanosoma cruzi from different areas of Argentina were classified into 12 zymodemes. The isolates were obtained from human patients with acute, chronic or congenital Chagas' disease, vector insects, domestic and sylvatic animals. Two out of 8 isolates cloned were shown to be heterogeneous. Zymodemes 1 and 12 exhibit widespread geographic distribution; isolates belonging to both zymodemes account for 55% of the total analyzed. The other zymodemes are not widely geographically dispersed. Although there is a clear predominance of zymodeme 1 among asymptomatic patients, the data do not show a clear relationship between particular zymodemes and the clinical picture. The results suggest that the sylvatic and domestic transmission cycles overlap. This remarkable heterogeneity of T. cruzi in Argentina supports the possible multiclonal origin of these parasite populations.

Animals↗
PubMed2002

Development and applications of transgenesis in the yellow fever mosquito, Aedes aegypti.

Transgenesis technology has been developed for the yellow fever mosquito, Aedes aegypti. Successful integration of exogenous DNA into the germline of this mosquito has been achieved with the class II transposable elements, Hermes, mariner and piggyBac. A number of marker genes, including the cinnabar(+) gene of Drosophila melanogaster, and fluorescent protein genes, can be used to monitor the insertion of these elements. The availability of multiple elements and marker genes provides a powerful set of tools to investigate basic biological properties of this vector insect, as well as the materials for developing novel, genetics-based, control strategies for the transmission of disease.

Aedes↗
PubMed2000

Tsetse--A haven for microorganisms.

Arthropods are involved in the transmission of parasitic and viral agents that cause devastating diseases in animals and plants. Effective control strategies for many of these diseases still rely on the elimination or reduction of vector insect populations. In addition to these pathogenic organisms, arthropods are rich in microbes that are symbiotic in their associations and are often necessary for the fecundity and viability of their hosts. Because the viability of the host often depends on these obligate symbionts, and because these organisms often live in close proximity to disease-causing pathogens, they have been of interest to applied biologists as a potential means to genetically manipulate populations of pest species. As knowledge on these symbiotic associations accumulates from distantly related insect taxa, conserved mechanisms for their transmission and evolutionary histories are beginning to emerge. Here, Serap Aksoy summarizes current knowledge on the functional and evolutionary biology of the multiple symbionts harbored in the medically and agriculturally important insect group, tsetse, and their potential role in the control of trypanosomiasis.

Animals↗
PubMed1993

Genetic structure of Trypanosoma cruzi populations from Argentina estimated from enzyme polymorphism.

Isolates of Trypanosoma cruzi from human patients, domestic and sylvatic animals and vector insects were obtained in different areas of Argentina. Electrophoretic patterns of enzymes from extracts of 95 isolates were analysed. On the basis of zymograms providing information on 10 loci, 12 zymodemes are described according to their genotypes. Data presented show fixed heterozygosity, absence of segregation of genotypes, significant departures from Hardy-Weinberg equilibrium, and over-represented genotypes. This evidence supports the hypothesis that sexual reproduction is very restricted or absent in this parasite. The proportion of polymorphic loci is 80%. The expected mean heterozygosity per locus (He) is 0.43, while the observed value (Ho) is 0.24. Differences between these values may be explained by accepting a basically clonal structure for T. cruzi. The data matrix of 12 zymodemes using 28 characters was analysed using a Wagner parsimony algorithm. Two equally most parsimonious unrooted trees were generated; both have 39 steps. The results show clusters clearly separated according to the geographical origin of the stocks. There are some indications of some correlations between genetic composition of the parasite and the clinical picture of the infection in human patients.

Animals↗
PubMed1998

Comparative kinetics of bloodmeal intake by Triatoma infestans and Rhodnius prolixus, the two principal vectors of Chagas disease.

Chagas disease vector insects Triatoma infestans and Rhodnius prolixus (fifteen stage III nymphs per 4 litre cage) were allowed to feed on anaesthetized mice for 1 h (control group), or on active non-anaesthetized mice (NAM) for 2, 4 or 8 h exposure. The bloodmeal size (weight increase) for both species was proportional to the duration of contact with NAM, due to ingestion of multiple small bloodmeals, up to 142% of control weight for T. infestans with 8 h exposure to NAM. The mean weight increase of T. infestans nymphs after 4 h contact with NAM was similar to that of the control group, whereas for R. prolixus, 8 h contact with NAM gave only 64% of the control value. For both species of insect, within 4 h of feeding, > 20% of the bloodmeal weight was lost by defaecation and diuresis. The proportions of unfed nymphs and mortality during 2 h contact with NAM were significantly higher for R. prolixus, demonstrating better exploitation of the host blood source by T. infestans, apparently because during blood-feeding the latter insect species caused less irritation to the host.

Animals↗
PubMed2004

Stages of infection during the tripartite interaction between Xenorhabdus nematophila, its nematode vector, and insect hosts.

Bacteria of the genus Xenorhabdus are mutually associated with entomopathogenic nematodes of the genus Steinernema and are pathogenic to a broad spectrum of insects. The nematodes act as vectors, transmitting the bacteria to insect larvae, which die within a few days of infection. We characterized the early stages of bacterial infection in the insects by constructing a constitutive green fluorescent protein (GFP)-labeled Xenorhabdus nematophila strain. We injected the GFP-labeled bacteria into insects and monitored infection. We found that the bacteria had an extracellular life cycle in the hemolymph and rapidly colonized the anterior midgut region in Spodoptera littoralis larvae. Electron microscopy showed that the bacteria occupied the extracellular matrix of connective tissues within the muscle layers of the Spodoptera midgut. We confirmed the existence of such a specific infection site in the natural route of infection by infesting Spodoptera littoralis larvae with nematodes harboring GFP-labeled Xenorhabdus. When the infective juvenile (IJ) nematodes reached the insect gut, the bacterial cells were rapidly released from the intestinal vesicle into the nematode intestine. Xenorhabdus began to escape from the anus of the nematodes when IJs were wedged in the insect intestinal wall toward the insect hemolymph. Following their release into the insect hemocoel, GFP-labeled bacteria were found only in the anterior midgut region and hemolymph of Spodoptera larvae. Comparative infection assays conducted with another insect, Locusta migratoria, also showed early bacterial colonization of connective tissues. This work shows that the extracellular matrix acts as a particular colonization site for X. nematophila within insects.

Animals↗