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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↗

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↗

The effect of isometamidium chloride on Trypanosoma vivax occurring within the insect vector (Glossina).

The effect of chemoprophylaxis on developing and mature Trypanosoma vivax in Glossina tachinoides and G. palpalis palpalis was evaluated. Newly emerged G. tachinoides and G.p. palpalis flies were infected with T. vivax by allowing them to feed on parasitaemic animals. Three experiments were conducted and in each the flies were divided into two groups. One group of infected flies was fed on animals treated 1 day previously with isometamidium (1 mg/kg body wt. in a 4% soln.) after which the surviving flies were dissected and examined for trypanosomes. The other group was fed on untreated animals. Out of a total number of 123 flies which fed on treated animals, none were found to be infected, while 51 of 127 flies which fed on untreated animals were infected. It was concluded that prophylactic treatment of animals with isometamidium would eliminate T. vivax infections from the insect vector. The potential significance of this finding in the control of trypanosomiasis in the field is discussed.

Animals↗

Heterogeneity of rice ragged stunt oryzavirus genome segment 9 and its segregation by insect vector transmission.

Genomic heterogeneity of genome segment 9 (S9) of rice ragged stunt virus (RRSV) was investigated and a point mutation was found to be responsible for an electrophoretic mobility shift of S9 on polyacrylamide gel electrophoresis (PAGE). A new form of S9 (S9L) which migrated slightly faster than natural S9 (S9U) had the same length with A-->C transversion at nt 843. Synthetic S9 with a C:G pair at nt 843 migrated slightly faster than that with an A:U pair. Therefore, we conclude that the single point mutation shifts the electrophoretic mobility. Using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP), we could detect S9U and S9L alone or mixture in insect vectors after acquisition as well as in infected rice plants.

Animals↗

Wound tumor virus polypeptide synthesis in productive noncytopathic infection of cultured insect vector cells.

Inoculation of the leafhopper cell line AC-20 with wound tumor virus resulted in a productive noncytopathic infection with no detectable alteration of cellular protein synthesis. Virus-specific polypeptide synthesis, detectable by 8 h postinoculation, increased in a linear fashion, reaching a peak (approximately 10 to 15% of total protein synthesis) by 48 h postinoculation. The rate of viral protein synthesis continued at this level for several days but declined, relative to cellular protein synthesis, as infected cells were passaged. By passage 10, the synthesis of viral polypeptides was reduced to a level approximately 5% of that observed at 48 h postinoculation. Viral protein synthesis was not stimulated by superinfection. Viral antigens and infectious virus persisted in the majority (greater than 90%) of cells in an infected culture even after more than 100 passages. The synthesis of wound tumor virus polypeptides in infected insect vector cells appears to be regulated in a coordinated and selective manner.

Animals↗

DNA sequence and RFLP data reflect geographical spread and relationships of Amylostereum areolatum and its insect vectors.

The white rot fungus, Amylostereum areolatum (Basidiomycetes), is best known for its symbiotic relationship with various siricid wood wasp species. In this study, the relationship between isolates of A. areolatum associated with two wood wasp species, Sirex noctilio and S. juvencus, are considered to identify possible intraspecific groups. Isolates from the northern (native) and southern (exotic) hemispheres are included to determine patterns of geographical spread and origin of introductions into the southern hemisphere. The phylogenetic relationships of these isolates to authentic isolates of A. chailletii, A. laevigatum and A. ferreum were also investigated. Sequence and restriction fragment length polymorphism (RFLP) analyses of the variable nuc-IGS-rDNA region provided markers to distinguish intraspecific groups within A. areolatum. Isolates of A. areolatum associated with S. noctilio and S. juvencus contained four heterogenic sequences in the DNA region analysed. These sequences occurred in one of five combinations in each isolate. Some of these sequences were unique to isolates of A. areolatum from either wasp species, while others were present in both groups. This shows the ancient and specialized evolutionary relationship that exists between these insects and fungi. Isolates from the southern hemisphere all share the same sequence group. This supports previous hypotheses that S. noctilio has spread between countries and continents of this region. At the interspecific level, the IGS-rDNA sequence analysis showed that A. ferreum and A. laevigatum are closely related to each other, and they in turn are related to A. chailletii. Amylostereum areolatum was the most distinctly defined species in the genus. This can be attributed to the obligate relationship between A. areolatum and its insect vectors. Polymerase chain reaction-RFLP analysis was also shown to be an effective tool to distinguish between the different species of Amylostereum.

Animals↗

Abortive infection of Lutzomyia longipalpis insect vectors by aflagellated LdARL-3A-Q70L overexpressing Leishmania amazonensis parasites.

Leishmania donovani ADP-ribosylation factor-like protein 3A (LdARL-3A) is a small G protein isolated from the protozoan parasite L. donovani with no defined physiological function. Previously [Cuvillier, A., Redon, F., Antoine, J.-C., Chardin, P., DeVos, T., and Merlin, G. (2000) J Cell Sci 113: 2065-2074] we have shown that overexpression in L. amazonensis promastigotes of the mutated protein LdARL-3A-Q70L, which remains constitutively associated with GTP, leads to the disappearance of the flagellum but does not impair cell viability or growth. Here we report that parasites overexpressing LdARL-3A-Q70L can invade in vitro cultivated macrophages to the same extent as control cells, demonstrating that the flagellum is not necessary for attachment to or engulfment into macrophages. These infections are productive because amastigotes differentiate and multiply. However, aflagellated LdARL-3A-Q70L-overexpressing Leishmania promastigotes could not survive in experimentally infected Lutzomyia longipalpis insect vectors, in contrast to untransfected or native LdARL-3A-overexpressing cells. Overexpression of the native and mutated proteins did not modify in vitro procyclic to metacyclic lipophosphoglycan maturation or differentiation from procyclic to metacyclic promastigotes, nevertheless there is a block in transmission of Leishmania. Better understanding of LdARL-3A pathways, notably those regarding flagellum biogenesis, may lead to the future development of Leishmania-specific drugs, which may stop parasite transmission in nature without affecting other species.

Animals↗