Observations on longevity and egg production in Aedes (Stegomyia) aegypti linn. mosquitoes infected with Uganda s virus.
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Expression of hepatitis B virus surface antigen gene in insect baculovirus vector systems.
The gene coding for the hepatitis B virus surface antigen (HBsAg) under the control of Autographa californicanuclear polyhedrosis virus polyhedrin promoter was successfully inserted into the genome of the Trichoplusia ni nuclear polyhdrosis virus. Infection of Spodoptera frugiperda cells with this recombinant virus produced a significant amount of HBsAg protein and secreted 22 nm particles containing the HBsAg. The expression of HBsAg gene was also obtained both in Trichoplusia ni larvae and in Philosamia cynthia ricini prepupae when infected with the recombinant virus. The HBsAg proteins expressed by baculovirus vector systems have morphological and antigenic properties identical to the 22 nm particles secreted by human cells.
Animals↗
Hematophagous insects as vectors for frog trypanosomes.
Experimental infections of three hematophagous arthropods (Rhodnius prolixus, Aedes aegypti, and Culex pipiens) with a trypanosome of the Trypanosoma rotatorium complex found in the frogs Hyla crepitans and Leptodactylus insularum revealed that A. aegypti is a good host for the flagellate; the course of development in the intestinal tract of the mosquito is described from 15 minutes to 168 hours. C. pipiens showed only low intestinal infections and R. prolixus did not permit development of the parasite. It is postulated that, in addition to the transmission of T. rotatorium by leeches, batrachophilic mosquitoes may transmit the parasite to frogs of more terrestrial habits by being ingested by these anurans.
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Approaches to vector control: new and trusted. 5. The epidemiological context of vector control.
This paper discusses 2 prominent, contemporary issues in the epidemiological context of insect vector control: (i) the magnitude of the control problem, and (ii) non-linear processes influencing vector control. It concludes that we still cannot reliably measure the scale of some important control problems; e.g., there is considerable uncertainty about the basic reproduction number of malaria. The emergence of new concepts such as strain-specific immunity, and a growing emphasis on disease control as distinct from infection control, mean that some quantitative problems are being redefined more quickly than they are being solved. Population biologists have urged exploration of density-dependent processes which may help or hinder new methods of vector control. This brief review of non-linear phenomena such as facilitation and limitation finds little evidence that they will significantly influence, e.g., the introduction of some novel refractory mechanism into a vector population.
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Culture-independent analysis of midgut microbiota in the arbovirus vector Culicoides sonorensis (Diptera: Ceratopogonidae).
Differences in midgut microbial communities inhabiting Culicoides spp., insect vectors of virus pathogens, may affect the variation observed in the ability of these biting midges to propagate arthropod-borne viruses. As a first step toward addressing this hypothesis, midgut bacterial communities were compared between Culicoides species expected to be efficient and inefficient vectors of virus pathogens. We used 16S rDNA sequence and restriction fragment information to provisionally identify 36 bacterial genera from guts of wild adult female biting midges, Culicoides sonorensis Wirth and Jones and Culicoides variipennis (Coquillet), from two geographical locations. Bacterial identification was made by sequence analysis of 16S rDNA fragments and by terminal restriction fragment length polymorphism analysis of polymerase chain reaction-amplified 16S rDNA fragments from adult guts. Of 36 bacterial genera identified, 12 had been previously identified in other insects: Comomonas, Enterobacter, Klebsiella, Acinetobacter, Pseudomonas, Stenotrophomonas, Staphylococcus, Chryseobacterium, Moraxella, Acholeplasma, Flavobacterium, and Rickettsia, Significant differences in bacterial community composition were found between all three groups of wild adult females analyzed: live-trapped C. sonorensis, laboratory-emerged C. sonorensis, and laboratory-emerged C. variipennis.
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Genetic variability in biochemical characters of Brazilian field populations of the Leishmania vector, Lutzomyia longipalpis (Diptera: Psychodidae).
The phlebotomine sand fly Lutzomyia longipalpis is the insect vector of visceral leishmaniasis, a protozoan disease of increasing incidence and distribution in Central and South America. Electrophoretic allele frequencies of 15 enzyme loci were compared among the L. longipalpis populations selected across its distribution range in Brazil. The mean heterozygosity of two colonized geographic strains (one each from Colombia and Brazil) were 6% and 13% respectively, with 1.6-1.9 alleles detected per locus. In contrast, among the seven widely separated field populations, the mean heterozygosity ranged from 11% to 16% with 2.1-2.9 alleles per locus. No locus was recovered that was diagnostic for any of the field populations. Allelic frequency differences among five field strains from the Amazon basin and eastern coastal Brazil were very low, with Nei's genetic distances of less than 0.01 separating them. The two inland and southerly samples from Minas Gerais (Lapinha) and Bahia (Jacobina) states were more distinctive with genetic distances of 0.024-0.038 and 0.038-0.059, respectively, when compared with the five other samples. These differences were the consequence of several high frequency alleles (glycerol-3-phosphate dehydrogenase [Gpd1.69] and phosphoglucomutase [Pgm1.69]) relatively uncommon in other strains. The low genetic distances, absence of diagnostic loci, and the distribution of genes in geographic space indicate L. longipalpis of Brazil to be a single, but genetically heterogeneous, polymorphic species.
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Gene vector and transposable element behavior in mosquitoes.
The development of efficient germ-line transformation technologies for mosquitoes has increased the ability of entomologists to find, isolate and analyze genes. The utility of the currently available systems will be determined by a number of factors including the behavior of the gene vectors during the initial integration event and their behavior after chromosomal integration. Post-integration behavior will determine whether the transposable elements being employed currently as primary gene vectors will be useful as gene-tagging and enhancer-trapping agents. The post-integration behavior of existing insect vectors has not been extensively examined. Mos1 is useful as a primary germ-line transformation vector in insects but is inefficiently remobilized in Drosophila melanogaster and Aedes aegypti. Hermes transforms D. melanogaster efficiently and can be remobilized in this species. This element is also useful for creating transgenic A. aegypti, but its mode of integration in mosquitoes results in the insertion of flanking plasmid DNA. Hermes can be remobilized in the soma of A. aegypti and transposes using a common cut-and-paste mechanism; however, the element does not remobilize in the germ line. piggyBac can be used to create transgenic mosquitoes and occasionally integrates using a mechanism other than a simple cut-and-paste mechanism. Preliminary data suggest that remobilization is infrequent. Minos also functions in mosquitoes and, like the other gene vectors, appears to remobilize inefficiently following integration. These results have implications for future gene vector development efforts and applications.
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Field prevalence of Wolbachia in the mosquito vector Aedes albopictus.
The endosymbiotic bacteria in the genus Wolbachia have been proposed as a potential candidate to deliver pathogen-blocking genes into natural populations of medically important insects. The successful application of Wolbachia in insect vector control depends on the ability of the agent to successfully invade and maintain itself at high frequency under field conditions. Here, we evaluated the prevalence of Wolbachia infections in a field population of the Wolbachia-superinfected mosquito Aedes albopictus. A field prevalence of 100% (n = 1,016) was found in a single population in eastern Thailand via polymerase chain reaction (PCR) testing of Wolbachia both from individual parent females and their corresponding F1 offspring. This is the first report of accurate Wolbachia prevalence in a field population of an insect disease vector. The prevalence of superinfection was estimated to be 99.41%. All single-infected individual mosquitoes (n = 6) were found to harbor group A Wolbachia. For this particular population, none was found to be single-infected with group B Wolbachia. Our results also show that PCR testing of field materials alone without checking F1 offspring overestimated the natural prevalence of single infection. Thus, the confirmation of infection status by means of F1 offspring was critical to the accurate estimates of Wolbachia prevalence under field conditions.
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[Standardization of conditions for PCR detection of Leishmania spp. DNA in sand flies (Diptera, Psychodidae)].
The correct identification of etiological agents in vector insects is crucial for epidemiological studies. Identification of flagellates in such vectors, usually by dissection of the digestive tract and microscopic observation of the contents as well as attempts at parasite isolation from insects in culture media, have proven operationally inadequate and with poor diagnostic specificity, since female sand flies are also hosts for other flagellates like Trypanosoma and Endotrypanum. Due to the efficiency and specificity of DNA target sequence amplification by polymerase chain reaction (PCR), the latter could be used to investigate the presence of Leishmania in sand flies, although the insects need to be properly stored and the Leishmania DNA extracted using appropriate methodology. This paper describes methodologies to standardize sand fly storage and Leishmania DNA extraction in such specimens as a more practical method in field studies.
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A general model for the African trypanosomiases.
A general mathematical model of a vector-borne disease involving two vertebrate host species and one insect vector species is described. The model is easily extended to other situations involving more than two hosts and one vector species. The model, which was developed from the single-host model for malaria described by Aron & May (1982), is applied to the African trypanosomiases and allows for incubation and immune periods in the two host species and for variable efficiency of transmission of different trypanosome species from the vertebrates to the vectors and vice versa. Equations are derived for equilibrium disease prevalence in each of the species involved. Model predictions are examined by 3-dimensional phase-plane analysis, which is presented as a simple extension of the 2-dimensional phase-plane analysis of the malaria model. Parameter values appropriate for the African trypanosomiases are derived from the literature, and a typical West African village situation is considered, with 300 humans, 50 domestic animals and an average population of 5000 tsetse flies. The model predicts equilibrium prevalences of Trypanosoma vivax, T. congolense and T. brucei of 47.0, 45.8 and 28.7% respectively in the animal hosts, 24.2, 3.4 and 0.15% in the tsetse vectors, and a 7.0% infection of humans with human-infective T. brucei. The contribution to the basic rate of reproduction of the human-infective T. brucei is only 0.11 from the human hosts and 2.54 from the animal hosts, indicating that in the situation modelled human sleeping sickness cannot be maintained in the human hosts alone. The animal reservoir is therefore crucial in determining not only the continued occurrence of the disease in humans, but its prevalence in these hosts as well. The effect of changing average fly density on equilibrium disease prevalences is examined, together with the effect of seasonal changes in fly numbers on disease incidence. In a seasonal situation changes in fly mortality rates affect both future population size and infection rate. Peak disease incidence lags behind peak fly numbers, and that in the less favoured host lags behind that in the more favoured host. Near the threshold fly density for disease transmission disease incidence is more changeable than at higher fly densities and may even exceed equilibrium prevalence at the same average fly density (because most hosts are susceptible at the time that fly numbers begin their annual increase).(ABSTRACT TRUNCATED AT 400 WORDS)
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Phytoplasma-plant interactions: effector-mediated host reprogramming, hormonal crosstalk, metabolic alterations and plant-mediated vector manipulation.
Phytoplasmas are wall-less, phloem-restricted bacterial pathogens that infect over 1,000 plant species, causing substantial losses in agriculture, horticulture, and forestry worldwide. Despite their reduced genomes and limited metabolic autonomy, these obligate parasites colonize diverse hosts through secreted effector proteins that extensively reprogram plant development, metabolism, immune signalling, and vector interactions. Advances in genomics, transcriptomics, proteomics, metabolomics, and functional studies have substantially clarified the molecular basis of phytoplasma pathogenicity and symptom development. This review synthesizes current understanding of phytoplasma-plant interactions, covering phytoplasma biology, genome evolution, and the infection cycle across plant and insect vector hosts. We examine the molecular functions of key effectors, SAP11, SAP54/PHYL1, SAP05, TENGU, SWP1, and recently identified virulence factors, focusing on how they target host transcription factors, phytohormone networks, protein degradation pathways, and immune responses to promote colonization and disease progression. We further discuss how phytoplasma infection disrupts phytohormone signalling, primary and secondary metabolism, and developmental programs to produce characteristic disease symptoms, with particular attention to pathogen-induced changes in host volatiles and nutritional quality that alter vector behaviour and enhance transmission. Finally, we summarize insights from multi-omics studies and emerging management strategies, including CRISPR-based genome editing, RNAi, rapid molecular diagnostics, resistant cultivars, microbiome-based approaches, and sustainable vector control, and highlight key knowledge gaps and priorities for developing effective, environmentally sustainable phytoplasma disease management.
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[Current approach to the study of cytotaxonomy of vectors].
In order to avoid the slowness of the classical analysis of polytene chromosomes in insect vectors of diseases, a programme of image analysis on computer has been perfected. In its actual form, it allows to obtain a real straight image of the chromosome, the curve of DNA optic density for all the bands and the possibility to bring all the obtained images to a same standard length. The whole of the operations takes between 3 and 5 minutes per chromosome. Then, the author describes a way of approach to complete this programme in order to obtain a comparison of the analysed chromosomes allowing the automatic location of inversions.
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Trapping of Tomato yellow leaf curl virus (TYLCV) and other plant viruses with a GroEL homologue from the whitefly Bemisia tabaci.
To avoid destruction in the haemolymph of their vector, many plant circulative viruses interact with GroEL homologues produced by insect endosymbiotic bacteria. We have exploited this phenomenon to devise tools allowing trapping of plant viruses by either GroEL purified from the whitefly Bemisia tabaci or by whitefly GroEL over-expressed in E. coli. PCR tubes or 96-well plates coated with a GroEL preparation were incubated with cleared sap of virus infected plant leaves or insect vectors. GroEL-bound viruses were then identified by PCR or RT-PCR using virus-specific primers or by ELISA with virus specific antibodies. In this way Tomato yellow leaf curl virus (TYLCV) - a whitefly-transmitted geminivirus - was detected in plant sap, in extracts of leaf squashes and in homogenates of individual viruliferous whiteflies. Anti-GroEL antibody prevented TYLCV binding to GroEL. GroEL-bound virus was also detected by ELISA. GroEL was much more potent in binding TYLCV than commercial anti-TYLCV antibodies. In addition to several other geminiviruses, these procedures allowed detecting a variety of RNA viruses such as Cucumber mosaic virus (CMV), Prune dwarf virus (PDV) and Tomato spotted wilt (TSWV), but not Potato virus X and Potato virus Y (PVX and PVY), Grapevine leafroll-associated viruses (GLRV) and Tobacco mosaic virus (TMV). Predictions pertaining to viruses that do, or do not bind to GroEL, and applications in plant virus diagnosis, are presented.
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Dominant lethality and insect population control.
Recent advances in insect genetic engineering have opened up new possibilities in the genetic control of insect vectors of human diseases. We outline the current state of the sterile insect technique and show how the use of engineered dominant lethals can greatly increase the effectiveness of this approach. We consider alternative strategies based on the use of conditional dominant lethals that are not always active in the environment.
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Anthrax in the Gambia: an epidemiological study.
Epidemiological data on 448 cases of human cutaneous anthrax from the Gambia showed that this particular strain of anthrax bacillus causes widespread morbidity and some mortality with, at the same time, subclinical infection. Analysis also showed that anthrax is not an occupationally related disease in the Gambia.The possibility of human-to-human spread, affecting all age groups and both sexes, by means of a communal toilet article was also shown. The fact that the strain is a good toxin producer but contains a weak antigen may have accounted for the repeated clinical infection and the fact that antibody titres were generally transient. Subclinical infection in animals was also found, particularly in sheep and goats, and also, with an unusually low mortality, in cows. Insect vectors were not excluded, but were unlikely. Vultures may spread the disease from village to village. Some possible public health and immunization procedures are discussed, with a view to containing this difficult problem in this part of west Africa.
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Vector-borne diseases and their control.
Control of the vector is usually a crucial factor in control programs for tropical diseases spread by insect vectors. Successful control programs aim at vulnerable points in the interactions between the vector, the reservoir host, the pathogen, the human host, and the environment. The objective is to prevent potential transmission, or interrupt actual transmission, by reducing the abundance, longevity, or host contact of the vector--whichever is most appropriate to the particular pathogen or disease and the local situation. The importance of individual assessment in the light of local conditions and a knowledge of the biology of the local vector is stressed. The vector-borne diseases discussed here are malaria, filariasis, arbovirus diseases, trypanosomiasis, leishmaniasis, plague and rickettsiosis.
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Identification of filarial larvae in vectors by DNA hybridization.
Infectivity rates of insect vectors are the best criteria by which to assess the transmission of filarial parasites and the efficacy of filariasis control programs. Currently available DNA probes can be used to estimate the proportion of vectors containing larvae of a given filarial species but provide no information on three other important variables in the transmission dynamics of filarial nematodes: the developmental stage, the location and the actual number of larvae that are present in the vector, all of which can be reliably determined by microscopy. However, species identification is often difficult and sometimes impossible by conventional microscopy, which requires morphologically intact specimens. DNA is tough and DNA probing can identify worms that are dead and that have lost morphologic integrity; it also permits multiple analyses of the same specimen with different probes and has the potential for simultaneous processing of very large numbers of samples. Here, Senaroth Dissonoyake and Willy Piessens outline the route to the development of species- and life cycle stage-specific DNA/RNA probing reagents, and simple, reliable and quantitative technologies that can supplement and ultimately replace microscopic dissection.
Journal Article↗
Malaria parasite and vector genomes: partners in crime.
The publication of the genome sequences of the malaria parasite Plasmodium falciparum and the insect vector Anopheles gambiae paves the way for scientists to study these organisms by using technologies developed to observe global changes in transcription and translation, as well as computational tools. Researchers are now able to investigate complex changes involved in development, growth and reaction to external factors. Given the medical importance of these organisms, much of this work is targeted on drug or insecticide discovery (including mechanisms of resistance to existing treatments), but the genome information also provides the opportunity to develop novel therapies.
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