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Failure of embryos from bluetongue infected cattle to transmit virus to susceptible recipients or their offspring.

Sixty heifers were infected with bluetongue virus (BTV) by the bites of the vector and by inoculation with insect origin virus. During the acute and convalescent stages of the infection, embryos were collected nonsurgically from these animals and washed according to the recommendations of the International Embryo Transfer Society (1). No BTV was isolated from 77 of these embryos when they were inoculated onto cell culture and into embryonating chicken eggs. There was no evidence of lateral BTV transmission when 231 of these embryos were transferred into susceptible recipients, nor was there evidence of vertical BTV transmission to the 88 calves resulting from these transfers. Another six donors that were assumed to have recovered from a natural infection of BTV, were added to the study to increase the probability of obtaining embryos from a persistently infected BTV carrier. However, it was determined later that these animals had not been infected with BTV but with the closely-related epizootic hemorrhagic disease virus (EHDV). Embryos were collected from these donors and washed as above. Neither BTV nor EHDV was isolated from 26 of these embryos by the inoculation of cell culture and embryonating chicken eggs. There was no evidence of lateral BTV or EHDV transmission to recipients of 15 of these embryos or of vertical BTV or EHDV transmission to the resulting 7 calves. However, two recipients of embryos from one of these donors developed antibodies to BTV 6 to 9 months after transfer. Passive antibodies to BTV were also detected in their calves. There is good evidence that these two recipients acquired BTV from natural exposure to infected insect vectors and not from the transferred embryos.

Journal Article↗

Symbiosis and attenuation.

Parasitic protozoa and bacteria transmitted by vector insects and ticks resist destruction in the intermediate host, possibly through the possession of antigens similar to those of the host. There is no evidence that symbiotic microorganisms have evolved similar eclipsed antigens. Symbiotes are protected from destruction by the natural lytic agents in the host, because they remain for the most part inside special cells that have lost the ability to recognize the symbiotes as "nonself." Metallic ions may play an important role in attenuation through their effect on cell membranes and on the release of lytic agents. The symbiotes of insects are useful and interesting subjects for studying attenuation.

Animals↗

Age-grading and growth of Wuchereria bancrofti (Filariidea: Onchocercidae) larvae by growth measurements and its use for estimating blood-meal intervals of its Polynesian vector Aedes polynesiensis (Diptera: Culicidae).

Growth in length and width of Wuchereria bancrofti (Filariidea: Onchocercidae) larvae developing in its Polynesian vector Aedes polynesiensis (Diptera: Culicidae) was analysed using a mathematical approach to objectively extract patterns. L1 had a U-shaped growth in length, while widths followed an S-shaped function. L2 had an S-shaped growth in length and width. Growth in length of L3 was also S-shaped, while widths had an asymptotic size following a period of rapid shrinkage. The greatest difference between length and width was in stage 3 where the length was over 75 times greater than the width. The ratio of length to width was approximately 50 for microfilariae and only 10 for the L1 ('sausage') stage. Characteristic mean length (and width) were approximately 280(7) microm for microfilariae, approximately 181 microm for L1 at their smallest, and approximately 1584(22) microm for L3 infective larvae. There was a great increase in length during stage 2 from approximately 322(27) to approximately 982(31) microm. Stage duration decreased with increasing temperature while growth rate increased, giving steeper growth curves. There was no effect of temperature on size, except for L3, which were shorter when mosquitoes were reared at higher temperature. It appears that larval growth is a continuous process from microfilariae to the young L3 stage, and continuously modifies the larval parasite aspect, even within each stage. Thus, information on larval shape may be used as an age indicator and in some cases, may give an estimation on time elapsed since infection of the vector. An important demographic parameter used in most mathematical models describing transmission of parasites by insect vectors is the length of the gonotrophic cycle of the vector, i.e. the time interval between two successive blood-meals. Usual methods for computing such a parameter are based on mark-recapture techniques. However, reliable estimates need substantial capture rates, which are not always possible. This paper presents another approach in which marked mosquitoes are those naturally infected by W. bancrofti. For one mosquito, the time since infection is simply the age of the developing larval parasite. Our method first expresses the age of larval parasite as a fraction of total development time (from microfilariae entering the vector to L3 larvae) using a regression model based on measurements of the parasite's length and width. This fraction of development is then converted to a chronological age since infection, using a back-calculation procedure involving ambient temperatures and growth rates of W. bancrofti larvae in the vector. The method is applied to wild caught Ae. polynesiensis in French Polynesia to compute the length of the gonotrophic cycle. This mosquito species comes to bite approximately 3, 6-7 and 9 days after a first infectious blood-meal. Then the length of the gonotrophic cycle may be of 3-4 days.

Aedes↗

Cross-reactivity of vector-borne metacyclic forms of Trypanosoma cruzi with mammalian and culture stages.

Metacyclic forms of Trypanosoma cruzi isolated from the hindgut of infected insect vectors (Rhodnius prolixus) were found to be immunologically cross-reactive with cultured epimastigote, amastigote, and metacyclic stages of the parasite as well as with bloodstream trypomastigote forms by direct agglutination and indirect immunofluorescence techniques. Sera specific for each of these forms of the parasite systematically yielded maximal antibody titers when measured against the homologous antigen, indicating that antigenic determinants are shared by all of the developmental forms used in this work. Supporting this conclusion were the significant reductions in anti-insect-derived metacyclic antibody titer caused by absorption with any of the other life stages of T. cruzi. These results are relevant to the potential use of laboratory-grown forms of T. cruzi in vaccination against a natural infection with this parasite.

Agglutination Tests↗

[Mediterranean visceral leishmaniasis: physiopathology].

INFANTILE LEISHMANIASIS: The protozoan parasites of the genus Leishmania are the causal agent of various diseases ranging from cutaneous lesions to fatal systemic diseases. In southern France, Leishmania infantum is an endemic species recognized as the causal agent of infantile leishmaniasis (Mediterranean visceral leishmaniasis). Little is known about the pathophysiology of the disease in humans, but models in mice may provide a new approach. INSECT VECTOR: Leishmania infantum are carried by sand flies. Antigenic modifications of the promastigote forms occur in these insects modifying lipophyosphoglycan (LPG) or glyocprotein gp63. Salivary gland lysates from the sand flies also enhance Leishmania infectivity. HUMAN INFESTATION: In humans, LPG and gp63 play a role in complement fixation, cell adhesion and resistance to complement-mediated lysis. Macrophage expression of class I and II major histocompatibility complex antigens is suppressed. T cells and interferon gamma are very important keys in the control of the parasite infection.

Adolescent↗

The Anopheles gambiae genome: next steps for malaria vector control.

Malaria remains a major public health problem that is made worse by poor implementation of control measures, and by the spread of drug- and insecticide-resistant parasites and vectors, respectively. Availability of the Anopheles gambiae genome sequence will accelerate identification and exploitation of new target genes in this insect vector. This provides unique opportunities to improve on existing vector control tools and to generate new tools within a global partnership. However, significant capacity needs to be built for investigators in disease-endemic countries to exploit the genome data. When integrated with existing strategies, the new tools will form an effective package for selective vector control in an effort to prevent mortality and morbidity due to malaria.

Animals↗

Transposable elements as population drive mechanisms: specification of critical parameter values.

With a view to the possible use of transposable elements (TEs) as a mechanism to drive genes into insect vector populations, we used a three-parameter density dependent growth equation to examine the critical parameter values that determine whether or not a mobile element will spread and become fixed in a finite diploid vector population. Populations were simulated with parameter values affecting size, reproductive rate, density-dependence, and transposition efficiency of the mobile element. Simulations indicated that an equilibrium was reached quickly, typically in < 50 generations. Even when initially present at < or = 1% of a large population, the mobile element spread quickly and became fixed if transposition efficiency was equal to unity and infertility caused by the element decreased reproductive capacity by as much as 45%. These results were insensitive to the values of basic wild type reproductive rates and density dependence, but population size, transposition efficiency of the element, reproductive rate individuals bearing TEs and initial ratio of TE-bearing to wild individuals modified the outcome. As population size and transposition efficiency decreased in value, TEs became fixed less easily. However, even in populations as small as n = 100, an element with a transposition efficiency > 0.75 that reduces fertility < 25% will become fixed when introduced at a frequency as low as 1% of the total population. These results are consistent with previously reported population genetics models. They suggest that engineered transposons with a wide range of properties may be used to drive genes, such as those for parasite resistance, into wild vector populations.

Animals↗

Using Drosophila as a model insect.

The fruit fly Drosophila melanogaster has become such a popular model organism for studying human disease that it is often described as a little person with wings. This view has been strengthened with the sequencing of the Drosophila genome and the discovery that 60% of human disease genes have homologues in the fruit fly. In this review, I discuss the approach of using Drosophila not only as a model for metazoans in general but as a model insect in particular. Specifically, I discuss recent work on the use of Drosophila to study the transmission of disease by insect vectors and to investigate insecticide function and development.

Animals↗

Impact of vector control on a dengue fever outbreak in Trinidad, West Indies, in 1998.

In 1998, Trinidad experienced its first major outbreak of dengue haemorrhagic fever. Data from the Trinidad Public Health Laboratory, the National Surveillance Unit and Insect Vector Control Division, Ministry of Health, Trinidad and Tobago were analysed to determine the impact of vector control measures on the dengue outbreak. Geographical Information Systems (GIS)/Global Positioning Systems (GPS) were used to map cases and to distinguish epidemiological clusters. The Aedes aegypti population densities were higher than the 5% transmission threshold in all counties. The spatial distribution of dengue fever cases was significantly correlated with the heavily populated east-west corridor in the north and several distinctly separate clusters in the western part of the island. The temporal distribution patterns showed significantly more dengue fever cases occurring during the rainy season than during the dry season. This study documents the importance of vector control in the prevention of dengue transmission since no vaccine is currently available, and emphasizes the urgent need to understand better the environmental factors which contribute to the proliferation of this disease vector Ae. aegypti.

Adult↗

Identification of species-specific DNA sequences in North American blackflies.

Blackflies are vectors for Onchocerca volvulus, a filarial parasite that infects tens of millions of people in Africa and Latin America. Different species of the insect vectors, varying greatly in their ability to transmit the infection, may populate overlapping geographical regions and be morphologically similar. A rapid and reliable method for distinguishing among blackfly species is presently not available. We have isolated cloned DNAs coding for repetitive sequences that can distinguish between two species of North American blackflies, Simulium pictipes and Simulium vittatum. These DNAs do not cross hybridize to three African blackfly species. The assay is very sensitive, less than 1/1000th of the amount of DNA in a single fly being sufficient for detection with a radioactive probe. Diagnosis with nonradioactive probes by a procedure that is amenable for use in the field was also demonstrated.

Animals↗

Study on phlebotomine sand fly (Diptera: Psychodidae) fauna in Belo Horizonte, state of Minas Gerais, Brazil.

A study on the phlebotomine sand fly fauna in Belo Horizonte city, state of Minas Gerais, Brazil, was carried out. From April 2001 to March 2003, monthly systematic collections were performed in three houses from each of the nine regions of the city, using CDC light traps for four consecutive days. The traps were set into the houses and in peridomestic areas totaling 54 traps. A number of 3871 sand fly specimens of the genera Lutzomyia and Brumptomyia were collected. Sixty eight percent of the specimens were L. longipalpis and 16% L. whitmani, insect vectors of visceral and American cutaneous leishmaniasis, respectively. Environmental factors such as temperature, humidity, and frequency of precipitation suggest that the number of insects increases after rainy periods. During the same period mentioned above, seasonal captures were carried out in parks and green areas of Belo Horizonte, using Shannon trap. A total of 579 phlebotomine sand flies were collected from which 398 (68.7%) were females with the predominance of L. whitmani and L. monticola. Those specimens were used for natural infection examination, by polymerase chain reaction. No Leishmania DNA was present in any of the specimens tested.

Animals↗

Simultaneous stable expression of neomycin phosphotransferase and green fluorescence protein genes in Trypanosoma cruzi.

The ribosomal RNA (rRNA) gene promoter was used to construct plasmid vectors that simultaneously express multiple exogenous genes in Trypanosoma cruzi. Vector pBSPANEO expresses neomycin phosphotransferase, and pPAGFPAN expresses both green fluorescent protein and neomycin phosphotransferase from a single promoter. Both vectors require the presence of the rRNA promoter for stable transfection; epimastigotes transfected with pPAGFPAN strongly fluoresced due to green fluorescent protein expression. Intact plasmids were rescued from the T. cruzi-transfected population after >8 mo of culture, indicating stable replication of these vectors. Vectors were integrated into the rRNA locus by homologous recombination and into other loci, presumably by illegitimate recombination. Parasites bearing tandem concatamers of plasmids were also found among the transfectants. Transfectants expressing green fluorescent protein showed a bright green fluorescence distributed throughout the cell. Fluorescence was also detected in amastigotes after infection of mammalian cells with transfected parasites, indicating that the rRNA promoter can drive efficient expression of these reporter genes in multiple life-cycle stages of the parasite. Expression of the heterologous genes was detected after passage in mice or in the insect vector. These vectors will be useful for the genetic dissection of T. cruzi biology and pathogenesis.

Animals↗

Density-dependent timing of defaecation by Rhodnius prolixus, and its implications for the transmission of Trypanosoma cruzi.

Transmission of Trypanosoma cruzi is crucially dependent on the timing of defaecation by their insect vectors. Experimental studies on Rhodnius prolixus nymphs revealed a negative correlation between blood meal weight and defaecation time. Bugs which fed to repletion defaecated on average 7 min after feeding, whereas bugs with interrupted feeds defaecated about 1 h later. As blood meal weight of triatomine bugs is density-dependent, these results suggest that the greatest risk of successful T. cruzi transmission would occur in recently colonized houses where the bug population is still increasing, or in houses recolonized after a vector control attempt.

Animals↗

Immune mechanisms in insects.

Understanding immune evasion by parasites in their insect vectors requires some understanding of the insect immune system. Until fairly recently, technical difficulties in handling cells and plasma hampered laboratory investigations into insect immunology, but modern techniques combined with a sound knowledge of insect physiology are now permitting rapid advances. Rather than discussing the many controversies, this review aims to point out current areas of research into cellular and 'humoral' mechanisms that might be followed up by parasitologists.

Journal Article↗

Arthropod vectors in the evolution of bunyaviruses.

Viruses from each genus of Bunyaviridae have preferential relationships to the arthropods of only one or two families, i.e. Bunyavirus to mosquitoes (Culicidae), Phlebovirus to sand flies (Psychodidae) and mosquitoes, Uukuvirus and Nairovirus to ticks (Ixodidae and Argasidae). An exception is genus Hantavirus not proven to be transmitted by vectors. Within the Bunyavirus genus 16 serogroups have been recognized on the basis of their antigenic relationship. Based on isolations from the nature each serogroup is preferentially linked with arthropod species (mostly mosquitoes) of one, or two genera. For 8 out of 16 serogroups Culex mosquitoes are the main insect vectors. Two serogroups are linked with Aedes mosquitoes, three with Anopheles mosquitoes. Aedeomyia mosquitoes, Culicoides bitting miges and Hyalomma ticks are vectors of one serogroup each. Evolutionary trends within the genus Bunyavirus and within the family Bunyaviridae can be recognized based on relationships of bunyaviruses to their arthropod vectors.

Animals↗