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Metabolic syndrome and obesity in an insect.

Dragonflies infected with noninvasive gregarine gut parasites (Apicomplexa: Eugregarinorida) [corrected] have reduced flight-muscle performance, an inability to metabolize lipid in their muscles, twofold-elevated hemolymph carbohydrate concentrations, and they accumulate fat in their thorax in a manner analogous to mammalian obesity. Gregarine infection is associated with inappropriate responses of hemolymph carbohydrate concentration to insulin and with chronic activation in the flight muscles of p38 MAP kinase, a signaling molecule involved in immune and stress responses. Short-term exposure to gregarine excretory/secretory products caused elevated blood carbohydrate and p38 MAPK activation in healthy individuals. These characteristics comprise a set of symptoms and processes that are known in mammals as metabolic syndrome but which have not previously been described in other animal taxa. In addition to expanding the known taxonomic breadth of metabolic disease, these results indicate that insects may be useful experimental models for studying its underlying biology and mechanisms.

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Conservation of a gliding motility and cell invasion machinery in Apicomplexan parasites.

Most Apicomplexan parasites, including the human pathogens Plasmodium, Toxoplasma, and Cryptosporidium, actively invade host cells and display gliding motility, both actions powered by parasite microfilaments. In Plasmodium sporozoites, thrombospondin-related anonymous protein (TRAP), a member of a group of Apicomplexan transmembrane proteins that have common adhesion domains, is necessary for gliding motility and infection of the vertebrate host. Here, we provide genetic evidence that TRAP is directly involved in a capping process that drives both sporozoite gliding and cell invasion. We also demonstrate that TRAP-related proteins in other Apicomplexa fulfill the same function and that their cytoplasmic tails interact with homologous partners in the respective parasite. Therefore, a mechanism of surface redistribution of TRAP-related proteins driving gliding locomotion and cell invasion is conserved among Apicomplexan parasites.

12E7 Antigen↗

Mechanisms underlying the evolution and maintenance of functionally heterogeneous 18S rRNA genes in Apicomplexans.

In many species of the protist phylum Apicomplexa, ribosomal RNA (rRNA) gene copies are structurally and functionally heterogeneous, owing to distinct requirements for rRNA-expression patterns at different developmental stages. The genomic mechanisms underlying the maintenance of this system over long-term evolutionary history are unclear. Therefore, the aim of this study was to investigate what processes underlie the long-term evolution of apicomplexan 18S genes in representative species. The results show that these genes evolve according to a birth-and-death model under strong purifying selection, thereby explaining how divergent 18S genes are generated over time while continuing to maintain their ability to produce fully functional rRNAs. In addition, it was found that Cryptosporidium parvum undergoes a rapid form of birth-and-death evolution that may facilitate host-specific adaptation, including that of type I and II strains found in humans. This represents the first case in which an rRNA gene family has been found to evolve under the birth-and-death model.

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ApiEST-DB: analyzing clustered EST data of the apicomplexan parasites.

ApiEST-DB (http://www.cbil.upenn.edu/paradbs-servlet/) provides integrated access to publicly available EST data from protozoan parasites in the phylum Apicomplexa. The database currently incorporates a total of nearly 100,000 ESTs from several parasite species of clinical and/or veterinary interest, including Eimeria tenella, Neospora caninum, Plasmodium falciparum, Sarcocystis neurona and Toxoplasma gondii. To facilitate analysis of these data, EST sequences were clustered and assembled to form consensus sequences for each organism, and these assemblies were then subjected to automated annotation via similarity searches against protein and domain databases. The underlying relational database infrastructure, Genomics Unified Schema (GUS), enables complex biologically based queries, facilitating validation of gene models, identification of alternative splicing, detection of single nucleotide polymorphisms, identification of stage-specific genes and recognition of phylogenetically conserved and phylogenetically restricted sequences.

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ApiDB: integrated resources for the apicomplexan bioinformatics resource center.

ApiDB (http://ApiDB.org) represents a unified entry point for the NIH-funded Apicomplexan Bioinformatics Resource Center (BRC) that integrates numerous database resources and multiple data types. The phylum Apicomplexa comprises numerous veterinary and medically important parasitic protozoa including human pathogenic species of the genera Cryptosporidium, Plasmodium and Toxoplasma. ApiDB serves not only as a database in its own right, but as a single web-based point of entry that unifies access to three major existing individual organism databases (PlasmoDB.org, ToxoDB.org and CryptoDB.org), and integrates these databases with data available from additional sources. Through the ApiDB site, users may pose queries and search all available apicomplexan data and tools, or they may visit individual component organism databases.

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Sex allocation and population structure in apicomplexan (protozoa) parasites.

Establishing the selfing, rate of parasites is important for studies in clinical and epidemiological medicine as well as evolutionary biology Sex allocation theory offers a relatively cheap and easy way to estimate selfing rates in natural parasite populations. Local mate competition (LMC) theory predicts that the optimal sex ratio (r*; defined as proportion males) is related to the selfing rate (s) by the equation r* = (1-s)/2. In this paper, we generalize the application of sex allocation theory across parasitic protozoa in the phylum Apicomplexa. This cosmopolitan phylum consists entirely of parasites, and includes a number of species of medical and veterinary importance. We suggest that LMC theory should apply to eimeriorin intestinal parasites. As predicted, data from 13 eimeriorin species showed a female-biased sex ratio, with the sex ratios suggesting high levels of selfing (0.8-1.0). Importantly, our estimate of the selfing rate in one of these species, Toxoplasma gondii, is in agreement with previous genetic analyses. In contrast, we predict that LMC theory will not apply to the groups in which syzygy occurs (adeleorins, gregarines and piroplasms). Syzygy occurs when a single male gametocyte and a single female gametocyte pair together physically or in close proximity, just prior to fertilization. As predicted, data from four adeleorin species showed sex ratios not significantly different from 0.5.

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Inbreeding and parasite sex ratios.

The breeding system of parasitic protozoa affects the evolution of drug resistance and virulence, and is relevant to disease diagnosis and the development of chemo- and immunotherapy. A major group of protozoan parasites, the phylum Apicomplexa, that includes the aetiological agents of malaria, toxoplasmosis and coccidiosis, all have dimorphic sexual stages. The sex ratio (proportion of males produced by parasites) is predicted to depend upon the inbreeding rate, and it has been suggested that sex-ratio data offer a relatively cheap and easy method for indirectly estimating inbreeding rates. Here, we exploit a new theoretical machinery to show that there are generally valid relationships between f, Wright's coefficient of inbreeding, and sex ratio, z(*), the generality being with respect to population structure. To focus the discussion, we concentrate on malaria and show that the previously derived result, f = 1 - 2z(*), does not depend on the artificial assumptions about population structure that were previously made. Not only does this justify the use of sex ratio as an indirect measure of f, but also we argue that it may actually be preferable to measure f by measuring sex ratios, rather than by measuring departures from Hardy-Weinberg genotypic proportions both in malaria and parasites more generally.

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The cell biology of secondary endosymbiosis--how parasites build, divide and segregate the apicoplast.

Protozoan parasites of the phylum Apicomplexa harbour a chloroplast-like organelle, the apicoplast. The biosynthetic pathways localized to this organelle are of cyanobacterial origin and therefore offer attractive targets for the development of new drugs for the treatment of malaria and toxoplasmosis. The apicoplast also provides a unique system to study the cell biology of endosymbiosis. This organelle is the product of secondary endosymbiosis, the marriage of an alga and an auxotrophic eukaryote. This origin has led to a fascinating set of novel cellular mechanisms that are clearly distinct from those employed by the plant chloroplast. Here we explore how the apicoplast interacts with its 'host' to secure building blocks for its biogenesis and how the organelle is divided and segregated during mitosis. Considerable advances in parasite genetics and genomics have transformed apicomplexans, long considered hard to study, into highly tractable model organisms. We discuss how these resources might be marshalled to develop a detailed mechanistic picture of apicoplast cell biology.

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Revision and checklist of the species (other than lecudina) of the aseptate gregarine family Lecudinidae.

A checklist is given of the 89 named species of the gregarine family Lecudininae, exclusive of the 42 named species of the genus Lecudina (phylum Apicomplexa, class Sporozoea, subclass Gregarinia, order Eugregarinida, suborder Aseptatina). The list includes also the synonyms, host names, locations in hosts, known geographic distributions of the species, as well as key references. Another list is given of synonyms, lapsi calami, nomina nuda, etc., associated with the genera. A new genus, Paraophioidina g.n., with type species, Paraophioidina haeckeli (Mingazzini, 1891) and a new species, Lankesteria ormieresi sp. n., are described. There are also new combinations in the genera Bhatiella, Ancora, Monocystella, Ascocystis, and Paraophioidina.

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Identification of an apically-located antigen that is conserved in sporozoan parasites.

Sporozoan parasites of the phylum Apicomplexa all possess common apical structures. The current study used a monoclonal antibody (mAb-E12) to identify a conserved antigen in the apical region of merozoites of seven species of Plasmodium (including rodent, primate and human pathogens), tachyzoites of Toxoplasma gondii, bradyzoites of Sarcocystis bovis, and sporozoites and merozoites of Eimeria tenella and E. acervulina. The antigen was also present in sporozoites of haemosporinid parasites. Immunofluorescence studies showed that the antigen was restricted to the apical 3rd of these invasive stages. Using immunoelectron microscopy, labeling was demonstrated in the region of the polar ring, below the paired inner membranes of the parasite pellicle, and near the subpellicular microtubules radiating from the polar ring of merozoites and sporozoites of E. tenella. The majority of the antigen could be extracted with 1% Triton-X 100, but a portion remained associated with the cytoskeletal elements. The molecule has a relative rate of migration (Mr) of 47,000 in Plasmodium spp. and 43-46,000 in coccidian species. Since the epitope recognized by mAb-E12 is highly conserved, restricted to motile stages, and appears to be associated with microtubules, this antigen could be involved in cellular motility and cellular invasion.

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Internal transcribed spacer dimorphism and diversity in Dientamoeba fragilis.

The internal transcribed spacer (ITS) region of the ribosomal RNA operon is frequently used for detecting sequence variation among closely related species as it is usually homogeneous within strains but evolves more rapidly than ribosomal RNA coding regions. We have studied this region in both genotypes of the human intestinal parasite Dientamoeba fragilis. In contrast to most organisms, we have identified extensive variation between copies of the sequence within the same strain. The ITS occurs in 2 major forms in each genotype but additional heterogeneity is also present within each form. The significance of this finding is unclear, but the only precedent for such variation is in the Apicomplexa, which have multiple dispersed ribosomal RNA operons in contrast to the tandem arrays found in most other eukaryotes.

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Cellular identity of a novel small subunit rDNA sequence clade of apicomplexans: description of the marine parasite Rhytidocystis polygordiae n. sp. (host: Polygordius sp., Polychaeta).

A new species of Rhytidocystis (Apicomplexa) is characterized from North American waters of the Atlantic Ocean using electron microscopy and phylogenetic analyses of small subunit (SSU) rDNA sequences. Rhytidocystis polygordiae n. sp. is a parasite of the polychaete Polygordius sp. and becomes the fourth described species within this genus. The trophozoites of R. polygordiae were relatively small oblong cells (L=35-55 microm; W=20-25 microm) and distinctive in possessing subterminal indentations at both ends of the cell. The surface of the trophozoites had six to eight longitudinal series of small transverse folds and several micropores arranged in short linear rows. The trophozoites of R. polygordiae were positioned beneath the brush border of the intestinal epithelium but appeared to reside between the epithelial cells within the extracellular matrix rather than within the cells. The trophozoites possessed a uniform distribution of paraglycogen granules, putative apicoplasts, mitochondria with tubular cristae, and a centrally positioned nucleus. The trophozoites were non-motile and lacked a mucron and an apical complex. Intracellular sporozoites of R. polygordiae had a conoid, a few rhoptries, micronemes, dense granules, and a posteriorly positioned nucleus. Phylogenies inferred from SSU rDNA sequences demonstrated a close relationship between R. polygordiae and the poorly known parasite reported from the hemolymph of the giant clam Tridacna crocea. The rhytidocystid clade diverged early in the apicomplexan radiation and showed a weak affinity to a clade consisting of cryptosporidian parasites, monocystids, and neogregarines.

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Comparative genome analysis reveals a conserved family of actin-like proteins in apicomplexan parasites.

BACKGROUND: The phylum Apicomplexa is an early-branching eukaryotic lineage that contains a number of important human and animal pathogens. Their complex life cycles and unique cytoskeletal features distinguish them from other model eukaryotes. Apicomplexans rely on actin-based motility for cell invasion, yet the regulation of this system remains largely unknown. Consequently, we focused our efforts on identifying actin-related proteins in the recently completed genomes of Toxoplasma gondii, Plasmodium spp., Cryptosporidium spp., and Theileria spp. RESULTS: Comparative genomic and phylogenetic studies of apicomplexan genomes reveals that most contain only a single conventional actin and yet they each have 8-10 additional actin-related proteins. Among these are a highly conserved Arp1 protein (likely part of a conserved dynactin complex), and Arp4 and Arp6 homologues (subunits of the chromatin-remodeling machinery). In contrast, apicomplexans lack canonical Arp2 or Arp3 proteins, suggesting they lost the Arp2/3 actin polymerization complex on their evolutionary path towards intracellular parasitism. Seven of these actin-like proteins (ALPs) are novel to apicomplexans. They show no phylogenetic associations to the known Arp groups and likely serve functions specific to this important group of intracellular parasites. CONCLUSION: The large diversity of actin-like proteins in apicomplexans suggests that the actin protein family has diverged to fulfill various roles in the unique biology of intracellular parasites. Conserved Arps likely participate in vesicular transport and gene expression, while apicomplexan-specific ALPs may control unique biological traits such as actin-based gliding motility.

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Secretory organelles of pathogenic protozoa.

Secretory processes play an important role on the biology and life cycles of parasitic protozoa. This review focus on basic aspects, from a cell biology perspective, of the secretion of (a) micronemes, rhoptries and dense granules in members of the Apicomplexa group, where these organelles are involved in the process of protozoan penetration into the host cell, survival within the parasitophorous vacuole and subsequent egress from the host cell, (b) the Maurer's cleft in Plasmodium, a structure involved in the secretion of proteins synthesized by the intravacuolar parasite and transported through vesicles to the erythrocyte surface, (c) the secretion of macromolecules into the flagellar pocket of trypanosomatids, and (d) the secretion of proteins which make the cyst wall of Giardia and Entamoeba, with the formation of encystation vesicles.

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Cell fractionation of parasitic protozoa: a review.

Cell fractionation, a methodological strategy for obtaining purified organelle preparations, has been applied successfully to parasitic protozoa by a number of investigators. Here we present and discuss the work of several groups that have obtained highly purified subcellular fractions from trypanosomatids, Apicomplexa and trichomonads, and whose work have added substantially to our knowledge of the cell biology of these parasites.

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A survey of hemoparasite infections in free-ranging mammals and reptiles in French Guiana.

Blood smears of 1,353 free-ranging mammals (35 species) and 112 reptiles (31 species) from French Guiana were examined for hemoparasites. Parasites from 3 major groups were recorded: Apicomplexa (including hemogregarines, piroplasms, and Plasmodium spp.), Trypanosomatidae, and Filaroidea. Fifty percent of the individuals (86% of the species) were infected by parasites from at least 1 group. Hemogregarines, identified as Hepatozoon sp., infected numerous snakes with high prevalences (30-100%); infection is reported for the first time in 5 host genera of snakes: Clelia, Oxybelis, Pseustes, Rhinobotryum, and Bothriopsis. Infections were also observed in 4 marsupial species and 1 rodent. Hepatozoon spp. recorded in Didelphis albiventris (Marsupialia) and Coendou prehensilis (Rodentia) may be new species. Plasmodium sp. were observed in 2 snake species, Dipsas indica (Colubridae) and Bothrops atrox (Viperidae). Plasmodium brasilianum was recorded in all 5 primate species examined. Piroplasms were observed in all mammal orders except primates. Large terrestrial rodents were the main hosts of members of the Babesidae; 42% of Myoprocta acouchy, 36% of Dasyprocta agouti, and 44% of Agouti paca were infected. Trypanosomes were common in mammals and were recorded in 70% of the examined genera. Trypanosoma cruzi-like infections were reported in 21 mammal species, including sloths, rodents, carnivores, and primates. Microfilariae were also widespread, with higher prevalences in sloths, anteaters, and porcupines (>40% of the individuals infected) and in tamarins (95% infected). This survey highlights some potential anthropozoonotic risks due to the recent further evidence of Plasmodium brasilianum and P. malariae as a single species and to the increased diversity of hosts for Trypanosoma cruzi.

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A molecular phylogeny of malarial parasites recovered from cytochrome b gene sequences.

A phylogeny of haemosporidian parasites (phylum Apicomplexa, family Plasmodiidae) was recovered using mitochondrial cytochrome b gene sequences from 52 species in 4 genera (Plasmodium, Hepatocystis, Haemoproteus, and Leucocytozoon), including parasite species infecting mammals, birds, and reptiles from over a wide geographic range. Leucocytozoon species emerged as an appropriate out-group for the other malarial parasites. Both parsimony and maximum-likelihood analyses produced similar phylogenetic trees. Life-history traits and parasite morphology, traditionally used as taxonomic characters, are largely phylogenetically uninformative. The Plasmodium and Hepatocystis species of mammalian hosts form 1 well-supported clade, and the Plasmodium and Haemoproteus species of birds and lizards form a second. Within this second clade, the relationships between taxa are more complex. Although jackknife support is weak, the Plasmodium of birds may form 1 clade and the Haemoproteus of birds another clade, but the parasites of lizards fall into several clusters, suggesting a more ancient and complex evolutionary history. The parasites currently placed within the genus Haemoproteus may not be monophyletic. Plasmodium falciparum of humans was not derived from an avian malarial ancestor and, except for its close sister species, P. reichenowi, is only distantly related to haemospordian parasites of all other mammals. Plasmodium is paraphyletic with respect to 2 other genera of malarial parasites, Haemoproteus and Hepatocystis. Explicit hypothesis testing supported these conclusions.

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Two new species of Xiphocephalus in Eleodes tricostata and Eleodes fusiformis (Coleoptera: Tenebrionidae: Eleodini) from the sandhills of western Nebraska.

Xiphocephalus triplogemmatus n. sp. and Xiphocephalus quadratogemmatus n. sp. (Apicomplexa: Eugregarinida) are described from Eleodes tricostata and Eleodes fusiformis (Coleoptera: Tenebrionidae), respectively, collected from Keith County in the sandhills of western Nebraska. Gamonts can be diagnosed with some confidence based on relative size and shape of the deutomerite, but these taxa are distinguished by differences in oocyst size, shape, and residuua number. Together with Xiphocephalus ellisi from Eleodes opacus in the same region, X. triplogemmatus and X. quadratogemmatus form a distinct Nearctic xiphocephalid group that is morphologically distinct from groups that occur in the Palearctic, Ethiopian, and Oriental regions.

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