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Intra-stadial tick-borne Thogoto virus (Orthomyxoviridae) transmission: accelerated arbovirus transmission triggered by host death.

Haematophagous insect vectors of arthropod-borne viruses (arboviruses) feed repeatedly. Consequently, they can transmit arboviruses to more than one host during the same developmental stage (intra-stadial transmission). By contrast, ixodid ticks generally feed only once at each parasitic stage (larva, nymph, and adult) and hence they have only one opportunity for tickborne virus transmission per stadium (inter-stadial transmission). Under natural conditions, tick-infested hosts may die (from disease or other causes) before the ticks have completed their long period of engorgement. A laboratory model was used to investigate the consequences of premature host death on tick-borne virus transmission. We report intra-stadial transmission of Thogoto virus by the nymphal, male, and female ticks of Rhipicephalus appendiculatus. Tick-borne Thogoto virus infection caused viraemia and death of hamsters before the nymphal and adult ticks feeding on them had completed engorgement. The resulting partially fed ticks were allowed to continue engorgement on new, uninfected hosts (interrupted feeding). During feeding on the subsequent hosts, they transmitted the virus intra-stadially to susceptible hosts (hamsters), and to uninfected co-feeding ticks on non-susceptible hosts (guinea-pigs). Intra-stadial transmission, mediated by interrupted feeding, may help explain outbreaks of rapid and fatal tick-borne viral diseases, and the epidemiology as well as evolution of virulence, in a susceptible host population. Additionally, intra-stadial transmission provides an increased risk of tick-borne pathogen transmission to humans and domestic animals during slaughter and game hunting.

Animals↗

Hepatitis C virus infection in Iceland: a recently introduced blood-borne disease.

This study demonstrates a very high prevalence of antibodies to hepatitis C virus among Icelandic intravenous (i.v.) drug users. Of 152 identified i.v. drug users 95 (63%) were shown to have antibodies to the hepatitis C virus. In contrast the seroprevalence in the general Icelandic population is low, (0.2%). Almost all cases of hepatitis C virus infection in Iceland are due to i.v. drug use or to use of infected blood or blood products. Sporadic cases with unexplained modes of transmission, a significant portion of hepatitis C infections elsewhere, are virtually non-existent in Iceland. The results of this study are consistent with the hypothesis that the sporadic community-acquired cases could be caused by blood transfer due to bites from insect vectors such as mosquitoes which are not found in Iceland.

Adolescent↗

Lytic activity and structural differences of amphipathic peptides derived from trialysin.

Trialysin is a pore-forming protein found in the saliva of Triatoma infestans (Hemiptera, Reduviidae), the insect vector of Chagas' disease. The protein is active against a broad range of cell types from bacteria to eukaryotic cells. Recognizing that the N-terminus of trialysin harbors the lytic motif [Amino, R., Martins, R. M., Procopio, J., Hirata, I. Y., Juliano, M. A., and Schenkman, S. (2002) J. Biol. Chem. 277, 6207-6213], we designed a set of peptides scanning this region to investigate the structural basis of its biological function. Peptides encompassing residues 1-32 (P6), 1-27 (P7), and 6-32 (P5) efficiently induced lysis of the protozoan parasite Trypanosoma cruzi and Escherichia coli in the 0.4-9.0 microM range, while much higher concentrations were required to cause hemolysis. Other more internal peptides, including peptide P2 (residues 21-47) and others up to residue 52, were less effective. P6 turned out to be the most active of all. P7 has a significantly higher activity than P5 against E. coli, while P5 has a hemolytic activity comparable to that of P6. CD spectroscopy showed that all tested peptides acquire a comparable helical content in solvent mixtures or in detergent micelles. The solution structure of P2 and P5-P7 was determined in a 30% trifluoroethanol/water mixture by nuclear magnetic resonance. All peptides exhibit a structure characterized by a central helical fold, and except for P2, which does not show a continuous hydrophobic surface, they are amphipathic. The structural models show that P5 and P7 extend their structural similarities with the most active peptide, P6, in either the C-terminus or the N-terminus. Amino acid substitutions in the N-terminus of P6 improved hemolysis but did not change the activity against T. cruzi. These results suggest that while amphipathicity is essential for the lytic activity, the selectivity of the active peptides for specific organisms appears to be associated with the structural features of their N- and C-termini.

Amino Acid Sequence↗

Cloning and expression in E. coli of the malarial sporozoite surface antigen gene from Plasmodium knowlesi.

The malarial sporozoite, the infective stage found in the salivary gland of the insect vector, bears highly immunogenic surface antigen(s). Repeated exposure to irradiated sporozoites induces protection against malaria in several host species, including man. Further, monoclonal antibodies that confer passive immunity react with the immunogenic surface determinants of different sporozoite species. One approach to prevent malaria, therefore, would be to produce a vaccine that induces high titres of circulating antibodies against the sporozoite surface determinant(s). However, production of such a vaccine has not been possible since sporozoites cannot be cultivated in vitro and, therefore, only limited amounts of surface antigen may be obtained. To overcome this problem, we have prepared mRNA from Plasmodium knowlesi-infected mosquitoes to construct a cDNA library. From this library we have isolated a clone that expresses the sporozoite surface antigen as a beta-lactamase fusion protein in the plasmid pBR322. This is the first potentially protective malarial antigen to be cloned by recombinant DNA technology.

Animals↗

Expression of a polypeptide containing a dipeptide repeat is confined to the insect stage of Trypanosoma brucei.

The protozoan parasite Trypanosoma brucei is transmitted between mammalian hosts by the tsetse fly (Glossina spp.). Trypanosomes ingested by the fly undergo a number of changes in the insect midgut during differentiation to procyclic forms. These include the loss of the variant specific glycoprotein (VSG) coat and the appearance of a common set of procyclic surface antigens. In order to investigate genes other than VSG genes which are expressed only at certain stages of the life cycle, the first cDNA specific to procyclic culture form trypanosomes (equivalent to the stage found in the insect midgut) has been characterized. The encoded polypeptide shows several characteristics of membrane proteins, but its most striking feature is the presence of a repetitive amino-acid sequence in which there are 22 tandem repeats of the dipeptide-Glu-Pro-. Related genes are also found in other trypanosome species and in leishmania. This gene shows many similarities to a number of surface antigen genes described in malaria and, more recently, Trypanosoma cruzi. This is the first example of a repetitive sequence in a parasite protein which is present only in the insect vector, and which therefore cannot be implicated in the mammalian host immune response.

Amino Acid Sequence↗

The crystal structure of bluetongue virus VP7.

Bluetongue virus (BTV), a representative of the orbivirus genus of the Reoviridae, is considerably larger (at 80 nm across), and structurally more complex, than any virus for which we have comprehensive structural information. Orbiviruses infect mammalian hosts through insect vectors and cause economically important diseases of domesticated animals. They possess a segmented double-stranded RNA genome within a capsid composed of four major types of polypeptide chains. An outer layer of VP2 and VP5 is removed as the virus enters the target cell, to leave an intact core within the cell. This core is 70 nm across and composed of 780 copies of VP7 (M(r) 38K) that, as trimers, form 260 'bristly' capsomeres clothing an inner scaffold constructed from VP3 (M(r) 103K). We report here the crystal structure of VP7 from BTV serotype 10, which reveals a molecular architecture not seen previously in viral structural proteins. Each subunit consists of two domains, one a beta-sandwich, the other a bundle of alpha-helices, and a short carboxy-terminal arm which might tie trimers together during capsid formation. A concentration of methionine residues at the core of the molecule could provide plasticity, relieving structural mismatches during assembly.

Amino Acid Sequence↗

Effect of insect-mediated dispersal on the genetic structure of postglacial water mite populations

Assaying population structure in species that differ in dispersal ability can help to determine whether population differentiation is dependent on the movement of individuals between populations. Here, allozyme variation is analysed in over 1100 individuals from nine species and two species complexes of Arrenurus water mites collected throughout north-eastern North America. As larvae, eight taxa are obligate parasites of winged adult insects that provide the primary opportunity for dispersal. Three additional species have lost the ability to parasitize insects and do not disperse in this manner. Consistent with the glaciated history of the region, very low allozyme heterozygosity was found in these taxa (Ho = 0.00-0.12), near panmixia in five out of seven species for which population differentiation was calculated and no patterns of isolation by distance over spatial scales up to several hundred kilometres. Nonetheless, in two out of three comparisons between sister species with and without parasitic larvae, parasitism was significantly associated with higher heterozygosity. Population differentiation could also be contrasted for two of these sister species pairs; in each case, lower estimates of FST were found in the mites able to disperse on insects. The statistical significance of these contrasts was dependent on the method used to estimate variance. At the scale of the genus, behavioural differences among insect vectors allows for broader hypotheses that relate water mite genetic diversity to dispersal ability. For the genus, rank correlations of dispersal ability with direct count heterozygosity (n = 11) and population differentiation (n = 7) were not significantly different from zero. These results are consistent with the hypothesis that allozyme population structure is primarily the result of historical patterns in these regions. However, comparisons between sister species suggest a limited role for dispersal in homogenizing populations genetically, even when drift-gene flow equilibrium has not been achieved.

Journal Article↗

Immunopathology of flavivirus infections.

With the recent emergence of the flavivirus, West Nile virus (WNV), in particular, the New York strain of Lineage I WNV in North America in 1999, there has been a significant increase in activity in neurotropic flavivirus research. These viruses cause encephalitis that can result in permanent neurological sequelae or death. Attempts to develop vaccines have made progress, but have been variably successful, despite considerable commercial underwriting. Thus, the discovery of ways and means to combat disease is no less urgent. As such, most recent work has been directed towards dissecting and understanding the pathogenesis of disease, as a way of informing possible approaches to abrogation or amelioration of illness. Whether inherent to flaviviruses or because humans are incidental, dead-end hosts, it is clear that these viruses interact with their human hosts in extremely complex ways. This occurs from the cellular level, at which infection must be established to produce disease, to its interaction with the adaptive immune response, which may result in its eradication, with or without immunopathological and consequent neurological sequelae. As human proximity to and contact with flavivirus insect vectors and amplifying hosts cannot practically be eliminated, our understanding of the pathogenesis of flavivirus-induced diseases, especially with regard to possible targets for treatment, is imperative.

Animals↗

Analysis of a cross between green and red fluorescent trypanosomes.

Trypanosoma brucei undergoes genetic exchange in its insect vector, but the mechanism is unknown and no one has yet seen the process. By crossing genetically engineered red and green fluorescent trypanosomes, we have been able to pinpoint the location of genetic exchange in the fly and search for intermediate stages. In experimental crosses of red and green parental trypanosomes, yellow hybrid trypanosomes first appeared in the fly salivary glands as early as 13 days after infection and were observed only in flies with a mixture of red and green trypanosomes in one or both salivary glands. Despite high numbers of flies with mixed infections, yellow trypanosomes were not detected in the fly midgut or proventriculus. The hybrid nature of yellow trypanosomes was confirmed by analysis of molecular karyotypes and microsatellite alleles. As well as yellow hybrids, hybrid trypanosomes with red, green or no fluorescence were also recovered from fly salivary glands. Analysis of microsatellite alleles in parental and progeny clones showed Mendelian inheritance. Our findings are consistent with the hypothesis that mating takes place between trypanosomes in the salivary glands of the fly before they attach to the salivary gland epithelium.

Animals↗

Efficacy of permethrin-impregnated uniforms in the prevention of cutaneous leishmaniasis in Iranian soldiers.

BACKGROUND: One approach for prevention of cutaneous leishmaniasis (CL) is the creation of a barrier between the insect vector (phlebotoms) and the host. Many but not all researchers claim that permethrin-impregnated uniforms are effective for prevention of CL. We determined the efficacy of permethrin-impregnated uniforms for prevention of CL in Iranian soldiers. METHODS: A total of 324 soldiers were randomized to two equal groups, of which 272 soldiers completed the study. In group A, 134 soldiers were issued permethrin-impregnated uniforms. In group B, 138 soldiers were issued uniforms washed in water. The soldiers wore uniforms day and night for 3 months, and were observed for an additional period of 6 months. All the soldiers remained in the leishmania-endemic area of Isfahan during the 3 months. RESULTS: Nine (6.5%) of 138 soldiers wearing control uniforms and six (4.4%) of 134 soldiers wearing permathrin-impregnated uniforms acquired CL. The difference between two groups was not statistically significant (P < 0.05). CONCLUSION: Permethrin-impregnated uniforms are not effective for the prevention of CL.

Adult↗

Evasion of innate and adaptive immunity by flaviviruses.

After a virus infects an animal, antiviral responses are generated that attempt to prevent dissemination. Interferons, antibody, complement, T and natural killer cells all contribute to the control and eradication of viral infections. Most flaviviruses, with the exception of some of the encephalitic viruses, cause acute disease and do not establish persistent infection. The outcome of flavivirus infection in an animal is determined by a balance between the speed of viral replication and spread, and the immune system response. Although many of the mechanistic details require further elucidation, flaviviruses have evolved specific tactics to evade the innate and adaptive immune response. A more thorough understanding of these principles could lead to improved models for viral pathogenesis and to strategies for the development of novel antiviral agents.

Animals↗

Interspecific evolution: microbial symbiosis, endosymbiosis and gene transfer.

Microbial symbioses are interesting in their own right and also serve as exemplary models to help biologists to understand two important symbioses in the evolutionary past of eukaryotic cells: the origins of chloroplasts and mitochondria. Most, if not all, microbial symbioses have a chemical basis: compounds produced by one partner are useful for the other. But symbioses can also entail the transfer of genes from one partner to the other, which in some cases cements two cells into a bipartite, co-evolving unit. Here, we discuss some microbial symbioses in which progress is being made in uncovering the nature of symbiotic interactions: anaerobic methane-oxidizing consortia, marine worms that possess endosymbionts instead of a digestive tract, amino acid-producing endosymbionts of aphids, prokaryotic endosymbionts living within a prokaryotic host within mealybugs, endosymbionts of an insect vector of human disease and a photosynthetic sea slug that steals chloroplasts from algae. In the case of chloroplasts and mitochondria, examples of recent and ancient gene transfer to the chromosomes of their host cell illustrate the process of genetic merger in the wake of organelle origins.

Animals↗

Lectins (hemagglutinins) in the gut of the important disease vectors.

The review is devoted to the gut lectins/hemagglutinins of the following representatives of important disease vectors: ticks, kissing-bugs, mosquitoes, sandflies and tsetse flies. The paper surveys the recent knowledge on these carbohydrate binding factors with respect to their structural and functional properties, and their significance for pathogen/parasite transmission by the blood-sucking arthropods. Recent results suggest that in most vectors the gut lectin activities are blood-meal enhanced, might participate in blood-meal processing and digestion and could serve as antibacterial and antiparasitic agents.

Animals↗