Phospholipid composition ofDipylidium caninum

Summary The phospholipid composition ofDipylidium caninum has been studied. Chloroform-methanol-soluble fraction amounted to 2.4% and phospholipids to 0.5% of the wet weight of the parasite. Phosphatidyl choline and phosphatidyl ethanolamine represented the bulk of the phospholipids, whereas phosphatidyl serine, phosphatidyl inositol, lysolecithin and lysophosphatidyl ethanolamine were present in minor amounts. Sulfatides were also identified in this parasite.     

Babesia bovis: the effect of acute inflammation and isoantibody production in the detection of babesial antigens

Summary Immunoblots ofBabesia bovis antigen contain dominant antigens which react not only with antisera toB. bovis but with sera from naive calves recovering from an acute inflammatory reaction. It seems likely these antigens are from the host rather than the parasite.     

Evolution and ecological correlates of uniparental reproduction in freshwater snails

We review the spatial and temporal correlates of uniparental reproduction in freshwater snails as they pertain to the ecological hypotheses for the maintenance of biparental sex. The biogeographic evidence from two species (Potamopyrgus antipodarum andBulinus truncatus) presently supports the Red Queen hypothesis that biparental reproduction is selected as a way to reduce the risk to progeny of parasite attack. Uniparental reproduction in these species is associated with low levels of infection by parasites (castrating digenetic trematodes), suggesting that parthenogenesis or self-fertilization can replace cross-fertilization when the risk of infection is low. In addition, inB. truncatus, the opportunity for cross-fertilization coincides with the season in which parasite attack is highest. In a third species (Campeloma decisum), parthenogenetic reproduction is correlated with latitude and the presence of a non-castrating trematode that may prevent cross-fertilization; these patterns suggest that parthenogenesis has been selected as a mechanism to assure reproduction. Finally, we discuss the spotty taxonomic distribution of parthenogenetic species.     

Presence of anti-Trypanosoma cruzi antibodies in the sera of mice with experimental autoimmune myocarditis

Summary The existence of antigens shared in common byT. cruzi and heart muscle cells is suggested by the presence of antibodies binding to the parasite surface in the serum of mice with autoimmune myocarditis induced by immunization with syngeneic heart antigens.     

Pathogenesis of Plasmodium falciparum malaria: the roles of parasite adhesion and antigenic variation

Malaria results in up to 2.5 million deaths annually, with young children and pregnant women at greatest risk. The great majority of severe disease is caused by Plasmodium falciparum. A characteristic feature of infection with P. falciparum is the accumulation or sequestration of parasite-infected red blood cells (RBCs) in various organs, such as the brain, lung and placenta, and together with other factors is important in the pathogenesis of severe forms of malaria. Sequestration results from adhesive interactions between parasite-derived proteins expressed on the surface of infected RBCs and a number of host molecules on the surface of endothelial cells, placental cells and uninfected RBCs. Some receptors for parasite adhesion have been implicated in particular malaria syndromes, such as intercellular adhesion molecule 1 in cerebral malaria and chondroitin sulfate A and hyaluronic acid in placental infection. The principal parasite ligand and antigen on the RBC surface, P. falciparum erythrocyte membrane protein 1 encoded by a multigene family termed var, is clonally variant, enabling evasion of specific immune responses. An understanding of these host-parasite interactions in the context of clinical disease and immunity may reveal potential targets to prevent or treat severe forms of malaria. Received 25 June 2001; received after revision 22 August 2001; accepted 24 August 2001     

Interaction of benznidazole reactive metabolites with nuclear and kinetoplastic DNA,proteins and lipids fromTrypanosoma cruzi

Summary Epimastigotes ofTrypanosoma cruzi (Tulahuen strain Tul 0 stock) biotransform benznidazole (N-benzyl-2-nitro-1-imidazone acetamide) to reactive metabolites that bind covalently to DNA, proteins and lipids of the parasite. These effects might be related to the trypanocidal action of benznidazole, a chemotherapeutic agent against Chagas' disease.     

1-(Substituted)benzyl-5-aminoimidazole-4-carboxamides are potent orally active inhibitors ofTrypanosoma cruzi in mice

Summary 1-(Substituted)benzyl-5-aminoimidazole-4-carboxamides are potent orally active inhibitors ofTrypanosoma cruzi infections in mice. The most active compounds are the 1-(4-chlorobenzyl)- and 1-(3,4-dichlorobenzyl)-analogs (L-153,094 [2] and L-153,153 [4], resp.) which are approximately 7-fold more potent upon oral administration than nifurtimox (Lampit) in suppressing parasite levels in the blood of mice with acuteTrypanosoma cruzi infections.     

Mast cells enter a teleost's brain by Xth cranial nerve in response toDiplostomum phoxini (Trematoda)

Summary Experimental infection byDiplostomum phoxini of the brain of laboratory hatched and rearedPhoxinus phoxinus induces the migration along the Xth cranial nerve of periodic acid Schiff positive granular leucocytes (PAS-GLs). These differentiate and grow into cells that lie between the parasite and the neurones of the host. The transformed cells are associated with elevated levels of heparin and serotonin in the brain; little histamine was detected. The reactive cell is identified as a mast cell.This work was carried out with M.R.L. Johnston's guidance. Teresa Townshend is thanked for assistance with techniques to hatch and raiseP. phoxinus.     

Arginase expression in peritoneal macrophages and increase in circulating polyamine levels in mice infected with Schistosoma mansoni

We investigated the nitric oxide (NO) synthase and arginase pathways in resident peritoneal macrophages of mice infected with the tropical parasite Schistosoma mansoni. The two enzymes may have opposite effects, insofar as NO may be involved in the killing of the parasite whereas arginase may stimulate parasite growth via polyamine synthesis. We determined the effects of the infection on the expression and activity of the two enzymes in macrophages, before and after cytokine activation. Cells from infected mice expressed the hepatic type I arginase, whereas in control cells, the enzyme was expressed only after cytokine activation, as were NO synthase II and type II arginase in both groups of cells. Moreover, we found that in infected mice, arginase expression in macrophages was associated with a ten fold increase in the concentration of circulating ornithine-derived polyamines. This may be of pathological importance, since parasitic helminths are though to be dependent on their hosts for the uptake and interconversion of polyamines. Received 13 March 2001; received after revision 4 May 2001; accepted 7 June 2001     

Binding of antimalarial drugs to hemozoin fromPlasmodium berghei

Summary Chloroquine, quinacrine and mefloquine bind toPlasmodium berghei hemozoin, hemin, heme, protoporphyrin IX and protease digested methemoglobin. This binding may be the basis for drug accumulation and action in the parasite.Acknowledgments. We thank the World Health Organization, the National Research Council of Thailand and Mahidol University for financial support. Dr Prapin Wilairat's suggestions are appreciated.     

Enzymatic characterization of nine endoparasite species of small ermine moths (Yponomeutidae)

Summary Eight hymenopterous and 1 dipterous species, all endoparasitic in eggs, larvae, or pupae of small ermine moths (Yponomeuta) were investigated for their allozyme variation at 3–29 loci. The mean heterozygosity level of the hymenopterous species is one-third of that of the dipterous species. Zymogram patterns of the parasite larvae do not interfere with those of the host.     

Host-plant pyrrolizidine alkaloids inNyctemera annulata Boisduval: Their persistence through the life-cycle and transfer to a parasite

Summary Larvae ofNyctemera annulata Boisduval ingest pyrrolizidine alkaloids fromSenecio spathulatus A. Rich. which subsequently appear in the adult months and their eggs; the acquisition of the alkaloids by a parasite of theN. annulata larvae provides a further illustration of their ability to persist in insects.Acknowledgments. We thank Dr R. Yamdagni of the Chemistry Department, University of Calgary, Canada, and Dr O. Madsen of the Organic Chemistry Department, The Technical University of Denmark, Lyngby, Denmark, for much help with the MS data collection. We are also very grateful to Dr C.C.J. Culvenor, CSIRO Animal Health Lab., Parkville, Victoria, Australia, who most generously provided us with authentic reference specimens of pyrrolizidine alkaloids.     

<Emphasis Type="Italic">Trichomonas vaginalis</Emphasis> degrades nitric oxide and expresses a flavorubredoxin-like protein: a new pathogenic mechanism?

Besides possessing many physiological roles, nitric oxide (NO) produced by the immune system in infectious diseases has antimicrobial effects. Trichomoniasis, the most widespread non-viral sexually transmitted disease caused by the microaerophilic protist Trichomonas vaginalis, often evolves into a chronic infection, with the parasite able to survive in the microaerobic, NO-enriched vaginal environment. We relate this property to the finding that T. vaginalis degrades NO under anaerobic conditions, as assessed amperometrically. This activity, which is maximal (133 ± 41 nmol NO/10⁸ cells per minute at 20°C) at low NO concentrations ( 1.2 M), was found to be: (i) NADH dependent, (ii) cyanide insensitive and (iii) inhibited by O₂. These features are consistent with those of the Escherichia coli A-type flavoprotein (ATF), recently discovered to be endowed with NO reductase activity. Using antibodies against the ATF from E. coli, a protein band was immunodetected in the parasite grown in a standard medium. If confirmed, the expression of an ATF in eukaryotes suggests that the genes coding for ATFs were transferred during evolution from anaerobic Prokarya to pathogenic protists, to increase their fitness for the microaerobic, parasitic life style. Thus the demonstration of an ATF in T. vaginalis would appear relevant to both pathology and evolutionary biology. Interestingly, genomic analysis has recently demonstrated that Giardia intestinalis and other pathogenic protists have genes coding for ATFs.Received 1 November 2003; received after revision 5 January 2004; accepted 13 January 2004     

<Emphasis Type="Italic">Plasmodium falciparum</Emphasis> possesses a unique dual-specificity serine/threonine and tyrosine kinase,Pfnek3

Despite the absence of classical tyrosine kinases encrypted in the kinome of Plasmodium falciparum, biochemical analyses have detected significant tyrosine phosphorylation in its cell lysates. Supporting such phosphorylation is critical for parasite development. These observations have thus raised queries regarding the plasmodial enzymes accountable for tyrosine kinase activities in vivo. In the current investigation, immunoblot analysis intriguingly demonstrated that Pfnek3, a plasmodial mitogen-activated protein kinase kinase (MAPKK), displayed both serine/threonine and tyrosine kinase activities in autophosphorylation reactions as well as in phosphorylation of the exogenous myelin basic protein substrate. The results obtained strongly support Pfnek3 as a novel dual-specificity kinase of the malarial parasite, even though it displays a HGDLKSTN motif in the catalytic loop that resembles the consensus HRDLKxxN signature found in the serine/threonine kinases. Notably, its serine/threonine and tyrosine kinase activities were found to be distinctly influenced by Mg²⁺ and Mn²⁺ cofactors. Further probing into the regulatory mechanism of Pfnek3 also revealed tyrosine phosphorylation to be a crucial factor that stimulates its kinase activity. Through biocomputational analyses and functional assays, tyrosine residues Y117, Y122, Y172, and Y238 were proposed as phosphorylation sites essential for mediating the catalytic activities of Pfnek3. The discovery of Pfnek3’s dual role in phosphorylation marks its importance in closing the loop for cellular regulation in P. falciparum, which remains elusive to date.     

Subcellular location of glycolytic enzymes inTrypanosoma brucei culture form

Summary The presence of a glycolytic complex or particle has been demonstrated in the insect midgut form ofTrypanosoma brucei brucei. It differs from the ‘glycosome’ of the bloodstream form of the parasite in that 3-phosphoglycerate kinase is absent. The latter enzyme appears to be cytosolic. Acknowledgment. We are grateful to Dr R. Brun for the kind supply of the strain. The work was funded by the Swiss National Science Foundation No. 3.331.0.78.     

On the phospholipids of adultAncylostoma caninum

Summary The phospholipid composition ofAncylostoma caninum has been examined. Phospholipids amounted to 0.9% of the wet weight of the parasite. Ethanolamine and choline containing lipids comprised about 76% of the total phospholipids. Lysolecithin, sphingomyelin, phosphatidyl inositol and phosphatidyl serine were present in minor amounts. Cerebrosides and sulfatides were also identified.Acknowledgements. We wish to acknowledge the contributions of Mr Ashok Chopra in the initial part of this work. Thanks are due to Mr Adarsh Kumar for skilled assistance.     

Hemagglutination of neuraminidase-treated human erythrocytes byLeishmania enriettii infected guinea-pig sera

Summary Sera from guinea-pigs infected with the protozoan parasiteLeishmania enriettii showed higher hemagglutination (HA) titres for neuraminidase treated human erythrocytes than those of normal guinea-pig sera. This HA activity was associated mostly with the 19S fraction of the immune serum and could be absorbed out with an antigenic fraction of the parasite membrane. Antigenic determinants involved in this HA reaction consisted of, at least, -d-galactosyl or lactosyl residues.     

Niche width of parasites in species-rich and species-poor communities

Summary Niche width of ectoparasites of marine fishes, as measured by host range and microhabitat width, is not affected by the number of species in a community. There is no reduction in species numbers of Monogenea due to greater numbers of other parasite species, and frequencies of infection with Monogenea are greater in tropical, species-rich communities.Supported by grants from the Australian Research Grants Committee and the University of New England. M. Heap provided valuable technical assistance, and F. Roubal made his data onAcanthopagrus australis available to me.     

Malaria vaccine

Summary Among infectious diseases caused by protozoa, malaria is still the greatest killer of children. Mortality in adults living in endemic areas is significantly lower because they frequently acquire partial or complete immunity to the major pathogen,Plasmodium falciparum. This natural protection indicates that vaccination may be possible, and the first candidate antigens were cloned with the use of human immune sera as probes. Genetic and biochemical analysis of the parasite proteins revealed that they are polymorphic, and frequently gene sequences were discovered which were specific for a particular parasite isolate, which eliminated most antigens for purposes of vaccine development. The most promising candidate antigens today are the major surface proteins of sporozoites and blood stage parasites. However, the immune response against those is not sufficient for complete protection, and additional, intensive research is necessary to identify new molecules to be included in a vaccine cocktail against malaria. The current spread of the disease due to increasing drug resistance of parasites and mosquito vectors emphasizes the urgent need for a vaccine.