A widespread coral-infecting apicomplexan with chlorophyll biosynthesis genes.

Apicomplexa is a group of obligate intracellular parasites that includes the causative agents of human diseases such as malaria and toxoplasmosis. Apicomplexans evolved from free-living phototrophic ancestors, but how this transition to parasitism occurred remains unknown. One potential clue lies in coral reefs, of which environmental DNA surveys have uncovered several lineages of uncharacterized basally branching apicomplexans1,2. Reef-building corals have a well-studied symbiotic relationship with photosynthetic Symbiodiniaceae dinoflagellates (for example, Symbiodinium3), but the identification of other key microbial symbionts of corals has proven to be challenging4,5. Here we use community surveys, genomics and microscopy analyses to identify an apicomplexan lineage-which we informally name 'corallicolids'-that was found at a high prevalence (over 80% of samples, 70% of genera) across all major groups of corals. Corallicolids were the second most abundant coral-associated microeukaryotes after the Symbiodiniaceae, and are therefore core members of the coral microbiome. In situ fluorescence and electron microscopy confirmed that corallicolids live intracellularly within the tissues of the coral gastric cavity, and that they possess apicomplexan ultrastructural features. We sequenced the genome of the corallicolid plastid, which lacked all genes for photosystem proteins; this indicates that corallicolids probably contain a non-photosynthetic plastid (an apicoplast6). However, the corallicolid plastid differs from all other known apicoplasts because it retains the four ancestral genes that are involved in chlorophyll biosynthesis. Corallicolids thus share characteristics with both their parasitic and their free-living relatives, which suggests that they are evolutionary intermediates and implies the existence of a unique biochemistry during the transition from phototrophy to parasitism.

The Organellar Genomes of Chromera and Vitrella, the Phototrophic Relatives of Apicomplexan Parasites.

Apicomplexa are known to contain greatly reduced organellar genomes. Their mitochondrial genome carries only three protein-coding genes, and their plastid genome is reduced to a 35-kb-long circle. The discovery of coral-endosymbiotic algae Chromera velia and Vitrella brassicaformis, which share a common ancestry with Apicomplexa, provided an opportunity to study possibly ancestral forms of organellar genomes, a unique glimpse into the evolutionary history of apicomplexan parasites. The structurally similar mitochondrial genomes of Chromera and Vitrella differ in gene content, which is reflected in the composition of their respiratory chains. Thus, Chromera lacks respiratory complexes I and III, whereas Vitrella and apicomplexan parasites are missing only complex I. Plastid genomes differ substantially between these algae, particularly in structure: The Chromera plastid genome is a linear, 120-kb molecule with large and divergent genes, whereas the plastid genome of Vitrella is a highly compact circle that is only 85 kb long but nonetheless contains more genes than that of Chromera. It appears that organellar genomes have already been reduced in free-living phototrophic ancestors of apicomplexan parasites, and such reduction is not associated with parasitism.

Morphology and Phylogenetic Position of Two New Gregarine Species (Apicomplexa: Eugregarinorida) Parasitizing the Lubber Grasshopper Taeniopoda centurio (Drury, 1770) (Insecta: Orthoptera: Romaleidae) in Mexico.

Eugregarines are understudied apicomplexan parasites of invertebrates inhabiting marine, freshwater, and terrestrial environments. Most currently known terrestrial eugregarines have been described parasitizing the gut from less than 1% of total insect diversity, with a high likelihood that the remaining insect species are infected. Eugregarine diversity in orthopterans (grasshoppers, locusts, katydids, and crickets) is still little known. We carried out a survey of the eugregarines parasitizing the Mexican lubber grasshopper, Taeniopoda centurio, an endemic species to the northwest of Mexico. We described two new eugregarine species from the gut of the host: Amoebogregarina taeniopoda n. sp. and Quadruspinospora mexicana n. sp. Both species are morphologically dissimilar in their life-cycle stages. Our SSU rDNA phylogenetic analysis showed that both species are phylogenetically distant to each other, even though they parasitize the same host. Amoebogregarina taeniopoda n. sp. clustered within the clade Gregarinoidea, being closely related to Amoebogregarina nigra from the grasshopper Melanoplus differentialis. Quadruspinospora mexicana n. sp. clustered within the clade Actinocephaloidea and grouped with Prismatospora evansi, a parasite from dragonfly naiads. Amoebogregarina taeniopoda n. sp. and Q. mexicana n. sp. represent the first record of eugregarines found to infect a species of the family Romaleidae.

Molecular systematics of marine gregarine apicomplexans from Pacific tunicates, with descriptions of five novel species of Lankesteria.

The eugregarines are a group of apicomplexan parasites that mostly infect the intestines of invertebrates. The high level of morphological variation found within and among species of eugregarines makes it difficult to find consistent and reliable traits that unite even closely related lineages. Based mostly on traits observed with light microscopy, the majority of described eugregarines from marine invertebrates has been classified into a single group, the Lecudinidae. Our understanding of the overall diversity and phylogenetic relationships of lecudinids is very poor, mainly because only a modest amount of exploratory research has been done on the group and very few species of lecudinids have been characterized at the molecular phylogenetic level. In an attempt to understand the diversity of marine gregarines better, we surveyed lecudinids that infect the intestines of Pacific ascidians (i.e. sea squirts) using ultrastructural and molecular phylogenetic approaches; currently, these species fall within one genus, Lankesteria. We collected lecudinid gregarines from six ascidian host species, and our data demonstrated that each host was infected by a different species of Lankesteria: (i) Lankesteria hesperidiiformis sp. nov., isolated from Distaplia occidentalis, (ii) Lankesteria metandrocarpae sp. nov., isolated from Metandrocarpa taylori, (iii) Lankesteria halocynthiae sp. nov., isolated from Halocynthia aurantium, (iv) Lankesteria herdmaniae sp. nov., isolated from Herdmania momus, (v) Lankesteria cf. ritterellae, isolated from Ritterella rubra, and (vi) Lankesteria didemni sp. nov., isolated from Didemnum vexillum. Visualization of the trophozoites with scanning electron microscopy showed that four of these species were covered with epicytic folds, whereas two of the species were covered with a dense pattern of epicytic knobs. The molecular phylogenetic data suggested that species of Lankesteria with surface knobs form a clade that is nested within a paraphyletic assemblage species of Lankesteria with epicytic folds.

Haemogregarines of freshwater turtles from Southeast Asia with a description of Haemogregarina sacaliae sp. n. and a redescription of Haemogregarina pellegrini Laveran and Pettit, 1910.

The uniform morphology of the developmental stages of Haemogregarina species and the insufficient information supplied by the simplistic descriptions of previous authors complicates their differential diagnosis and proper species identification. In this study, we detected Haemogregarina spp. in 6 out of 22 (27.2%) examined turtles originating from Southeast Asia, Malayemys subtrijuga (n = 4), Sacalia quadriocellata (n = 1) and Platysternon megacephalum (n = 1), and compared them with the available literature data. Microscopic analysis of our isolates distinguished 2 morphological species, Haemogregarina pellegrini and one new species, being described in this paper as Haemogregarina sacaliae sp. n. Phylogenetic analyses based on 1210 bp long fragment of 18S rDNA sequences placed both haemogregarines firmly within the monophyletic Haemogregarina clade. Isolates of H. pellegrini from 2 distantly related turtle hosts, M. subtrijuga and P. megacephalum, were genetically identical. Despite the fact that numerous Haemogregarina species of turtles have been described, the incompleteness of the morphological data and relatively low host specificity provides the space for large synonymy within this taxon. Therefore, a complex approach combining microscopic analyses together with molecular-genetic methods should represent the basic standard for all taxonomic studies.

Pacific marine gregarines (Apicomplexa) shed light on biogeographic speciation patterns and novel diversity among early apicomplexans.

Gregarines are the most biodiverse group of apicomplexan parasites. This group specializes on invertebrate hosts (e.g., ascidians, crustaceans, and polychaetes). Marine gregarines are of particular interest because they are considered to be the earliest evolving apicomplexan lineage, having subsequently speciated (and radiated) through virtually all existing animal groups. Still, mechanisms governing the broad (global) distribution and speciation patterns of apicomplexans are not well understood. The present study examines Pacific lecudinids, one of the most species-rich and diverse groups of marine gregarines. Here, marine polychaetes were collected from intertidal zones. Single trophozoite cells were isolated for light and electron microscopy, as well as molecular phylogenetic analyses using the partial 18S rRNA gene. The cytochrome c oxidase subunit 1 gene was used to confirm morphology-based host identification. This study introduces Undularius glycerae n. gen., n. sp. and Lecudina kitase n. sp. (Hokkaido, Japan), as well as Difficilina fasoliformis n. sp. (California, USA). Occurrences of Lecudina cf. longissima and Lecudina cf. tuzetae (California, USA) are also reported. Phylogenetic analysis revealed a close relationship between L. pellucida, L. tuzetae, and L. kitase n. sp. Additionally, clustering among North Atlantic and Pacific L. tuzetae formed a species complex, likely influenced by biogeography.

Apoptosis of Ascogregarina taiwanensis (Apicomplexa: Lecudinidae), which failed to migrate within its natural host.

Sexual reproduction of Ascogregarina taiwanensis (Apicomplexa: Lecudinidae), a parasite specific to the mosquito Aedes albopictus, in Malpighian tubules is initiated by the entry of the trophotozoites developed in the midgut shortly after pupation (usually <5 h). However, only a low proportion of trophozoites are able to migrate; others end up dying. In this study, we demonstrated that those trophozoites that failed to migrate eventually died of apoptosis. Morphological changes such as shrinkage, chromatin aggregations and formation of blunt ridges on the surface were seen in moribund trophozoites. In addition, DNA fragmentation of trophozoites isolated from the midgut of pupae was demonstrated by the presence of DNA ladders, Annexin V staining and TUNEL assays. Detection of caspase-like activity suggests that apoptosis of those trophozoites may have occurred through a mechanism of an intrinsic or mitochondrial-mediated pathway. Although apoptosis has been observed in various protozoan species, it is not clear how apoptosis in single-celled organisms might result from evolution by natural selection. However, we speculate that apoptosis may regulate the parasite load of A. taiwanensis within its natural mosquito host, leading to an optimized state of the survival rate for both parasite and host.

Molecular phylogeny of marine gregarine parasites (Apicomplexa) from tube-forming polychaetes (Sabellariidae, Cirratulidae and Serpulidae), including descriptions of two new species of Selenidium.

Selenidium is a genus of gregarine parasites that infect the intestines of marine invertebrates and have morphological, ecological, and motility traits inferred to reflect the early evolutionary history of apicomplexans. Because the overall diversity and phylogenetic position(s) of these species remain poorly understood, we performed a species discovery survey of Selenidium from tube-forming polychaetes. This survey uncovered five different morphotypes of trophozoites (feeding stages) living within the intestines of three different polychaete hosts. We acquired small subunit (SSU) rDNA sequences from single-cell (trophozoite) isolates, representing all five morphotypes that were also imaged with light and scanning electron microscopy. The combination of molecular, ecological, and morphological data provided evidence for four novel species of Selenidium, two of which were established in this study: Selenidium neosabellariae n. sp. and Selenidium sensimae n. sp. The trophozoites of these species differed from one another in the overall shape of the cell, the specific shape of the posterior end, the number and form of longitudinal striations, the presence/absence of transverse striations, and the position and shape of the nucleus. A fifth morphotype of Selenidium, isolated from the tube worm Dodecaceria concharum, was inferred to have been previously described as Selenidium cf. echinatum, based on general trophozoite morphology and host association. Phylogenetic analyses of the SSU rDNA sequences resulted in a robust clade of Selenidium species collected from tube-forming polychaetes, consisting of the two new species, the two additional morphotypes, S. cf. echinatum, and four previously described species (Selenidium serpulae, Selenidium boccardiellae, Selenidium idanthyrsae, and Selenidium cf. mesnili). Genetic distances between the SSU rDNA sequences in this clade distinguished closely related and potential cryptic species of Selenidium that were otherwise very similar in trophozoite morphology.

Discovery of a diverse clade of gregarine apicomplexans (Apicomplexa: Eugregarinorida) from Pacific eunicid and onuphid polychaetes, including descriptions of Paralecudina n. gen., Trichotokara japonica n. sp., and T. eunicae n. sp.

Marine gregarines are poorly understood apicomplexan parasites with large trophozoites that inhabit the body cavities of marine invertebrates. Two novel species of gregarines were discovered in polychaete hosts collected in Canada and Japan. The trophozoites of Trichotokara japonica n. sp. were oval to rhomboidal shaped, and covered with longitudinal epicytic folds with a density of six to eight folds/micron. The nucleus was situated in the middle of the cell, and the mucron was elongated and covered with hair-like projections; antler-like projections also extended from the anterior tip of the mucron. The distinctively large trophozoites of Trichotokara eunicae n. sp. lacked an elongated mucron and had a tadpole-like cell shape consisting of a bulbous anterior region and a tapered tail-like posterior region. The cell surface was covered with longitudinal epicytic folds with a density of three to five folds/micron. Small subunit (SSU) rDNA sequences of both species were very divergent and formed a strongly supported clade with the recently described species Trichotokara nothriae and an environmental sequence (AB275074). This phylogenetic context combined with the morphological features of T. eunicae n. sp. required us to amend the description for Trichotokara. The sister clade to the Trichotokara clade consisted of environmental sequences and Lecudina polymorpha, which also possesses densely packed epicyctic folds (3-5 folds/micron) and a prominently elongated mucron. This improved morphological and molecular phylogenetic context justified the establishment of Paralecudina (ex. Lecudina) polymorpha n. gen. et comb.

Four additional hepatozoon species (Apicomplexa: Hepatozoidae) from north Florida ratsnakes, genus Pantherophis.

Records from a colubrid host are reported for Hepatozoon horridus, described originally from a viperid snake. Hepatozoon horridus in Pantherophis guttatus (Colubridae) has gamonts 14-18.0 by 4.0-5.5 microm, with length by width (LW) 60-99 microm2, and L/W ratio 2.5-3.9. Spherical to elongate, usually ovoid oocysts with L/W ratio 1.0-3.7 contain 16-160 spherical to usually ovoid sporocysts 15-31 by 14-26 microm, with L/W ratio 1.0-1.4, that contain 5-24 sporozoites. Two additional Hepatozoon species are described from ratsnakes in north Florida. Hepatozoon quadrivittata n. sp. from Pantherophis obsoletus quadrivittatus has gamonts 12-17 by 4-6 microm, LW 56-102 microm2, and L/W ratio 2.6-3.8. Nearly spherical oocysts with L/W 1.0-1.1 contain 5-227 spherical to slightly ovoid sporocysts 20-48 by 19-45 microm, with L/W ratio 1.0-1.4, that contain 13-48 sporozoites. Hepatozoon spiloides n. sp. from Pantherophis obsoletus spiloides forms gamonts 12-15 by 4-5 microm with LW 48-75 microm2 and L/W ratio 2.6-3.5. Occasionally rounded but usually elongate oocysts, with L/W ratio 1.0-2.7, contain 5-21 spherical to elongate sporocysts 28-43 by 18-35 microm, L/W ratio 2.5-3.9. In the distinctive Hepatozoon sp. present in Pantherophis obsoletus spiloides, gamonts are 13-17 by 5-10 microm, with LW 75-140 microm2 and L/W ratio 1.4-3.0. Infected erythrocytes are always distorted and enlarged on average 2.5 times the size of uninfected cells, with nuclei enlarged by one-third and broadly elongated. Gamonts often stained deep blue, and cytoplasm of erythrocytes infected with mature gamonts was always dehemoglobinized. Sporogony could not be obtained in three feedings by hundreds of Aedes aegypti, which usually died within the first 24-48 hr.

Species boundaries in gregarine apicomplexan parasites: a case study-comparison of morphometric and molecular variability in Lecudina cf. tuzetae (Eugregarinorida, Lecudinidae).

Trophozoites of gregarine apicomplexans are large feeding cells with diverse morphologies that have played a prominent role in gregarine systematics. The range of variability in trophozoite shapes and sizes can be very high even within a single species depending on developmental stages and host environmental conditions; this makes the delimitation of different species of gregarines based on morphological criteria alone very difficult. Accordingly, comparisons of morphological variability and molecular variability in gregarines are necessary to provide a pragmatic framework for establishing species boundaries within this diverse and poorly understood group of parasites. We investigated the morphological and molecular variability present in the gregarine Lecudina cf. tuzetae from the intestines of Nereis vexillosa (Polychaeta) collected in two different locations in Canada. Three distinct morphotypes of trophozoites were identified and the small subunit (SSU) rDNA was sequenced either from multicell isolates of the same morphotype or from single cells. The aim of this investigation was to determine whether the different morphotypes and localities reflected phylogenetic relatedness as inferred from the SSU rDNA sequence data. Phylogenetic analyses of the SSU rDNA demonstrated that the new sequences did not cluster according to morphotype or locality and instead were intermingled within a strongly supported clade. A comparison of 1,657 bp from 45 new sequences demonstrated divergences between 0% and 3.9%. These data suggest that it is necessary to acquire both morphological and molecular data in order to effectively delimit the "clouds" of variation associated with each gregarine species and to unambiguously reidentify these species in the future.

Apicomplexa in mammalian cells: trafficking to the parasitophorous vacuole.

Most Apicomplexa reside and multiply in the cytoplasm of their host cell, within a parasitophorous vacuole (PV) originating from both parasite and host cell components. Trafficking of parasite-encoded proteins destined to membrane compartments beyond the confine of the parasite plasma membrane is a process that offers a rich territory to explore novel mechanisms of protein-membrane interactions. Here, we focus on the PVs formed by the asexual stages of two pathogens of medical importance, Plasmodium and Toxoplasma. We compare the PVs of both parasites, with a particular emphasis on their evolutionary divergent compartmentalization within the host cell. We also discuss the existence of peculiar export mechanisms and/or sorting determinants that are potentially involved in the post-secretory targeting of parasite proteins to the PV subcompartments.

Description of Trichotokara nothriae n. gen. et sp. (Apicomplexa, Lecudinidae)--an intestinal gregarine of Nothria conchylega (Polychaeta, Onuphidae).

The trophozoites of a novel gregarine apicomplexan, Trichotokara nothriae n. gen. et sp., were isolated and characterized from the intestines of the onuphid tubeworm Nothria conchylega (Polychaeta), collected at 20 m depth from the North-eastern Pacific Coast. The trophozoites were 50-155 microm long with a mid-cell indentation that formed two prominent bulges (anterior bulge, 14-48 microm wide; posterior bulge, 15-55 microm wide). Scanning electron microscopy (SEM) demonstrated that approximately 400 densely packed, longitudinal epicytic folds (5 folds/microm) inscribe the surface of the trophozoites, and a prominently elongated mucron (14-60 microm long and 6-12 microm wide) was covered with hair-like projections (mean length, 1.97 microm; mean width, 0.2 microm at the base). Although a septum occurred at the junction between the cell proper and the mucron in most trophozoites, light microscopy (LM) demonstrated that the cell proper extended into the core of the mucron as a thin prolongation. A spherical nucleus (8-20 microm) was situated in the middle of the trophozoites, and gamonts underwent caudal syzygy. The small subunit (SSU) rDNA sequence and molecular phylogenetic position of T. nothriae was also characterized. The sequence from this species was the most divergent of all SSU rDNA sequences currently known from gregarines and formed a weakly supported clade with Lecudina polymorpha, which also possesses densely packed epicyctic folds (3-5 folds/mum) and a prominently elongated mucron.

The life cycle and host specificity of Psychodiella sergenti n. sp. and Ps. tobbi n. sp. (Protozoa: Apicomplexa) in sand flies Phlebotomus sergenti and Ph. tobbi (Diptera: Psychodidae).

Two new gregarines in the recently erected genus Psychodiella (formerly Ascogregarina), Psychodiella sergenti n. sp. and Psychodiella tobbi n. sp., are described based on morphology and life cycle observations conducted on larvae and adults of their natural hosts, the sand flies Phlebotomus sergenti and Phlebotomus tobbi, respectively. The phylogenetic analyses inferred from small subunit ribosomal DNA (SSU rDNA) sequences indicate the monophyly of newly described species with Psychodiella chagasi. Ps. sergenti n. sp. and Ps. tobbi n. sp. significantly differ from each other in the life cycle and in the size of life stages. The sexual development of Ps. sergenti n. sp. (syzygy, formation of gametocysts and oocysts) takes place exclusively in blood-fed Ph. sergenti females, while the sexual development of Ps. tobbi n. sp. takes place also in males and unfed females of Ph. tobbi. The susceptibility of Phlebotomus perniciosus, Phlebotomus papatasi, Ph. sergenti, Ph. tobbi, and Phlebotomus arabicus to both gregarines was examined by exposing 1st instar larvae to parasite oocysts. High host specificity was observed, as both gregarines were able to fully develop and complete regularly the life cycle only in their natural hosts. Both gregarines are considered as serious pathogens in laboratory-reared colonies of Old World sand flies.

The Riveting Cellular Structures of Apicomplexan Parasites.

Parasitic protozoa of the phylum Apicomplexa cause a range of human and animal diseases. Their complex life cycles - often heteroxenous with sexual and asexual phases in different hosts - rely on elaborate cytoskeletal structures to enable morphogenesis and motility, organize cell division, and withstand diverse environmental forces. This review primarily focuses on studies using Toxoplasma gondii and Plasmodium spp. as the best studied apicomplexans; however, many cytoskeletal adaptations are broadly conserved and predate the emergence of the parasitic phylum. After decades cataloguing the constituents of such structures, a dynamic picture is emerging of the assembly and maintenance of apicomplexan cytoskeletons, illuminating how they template and orient critical processes during infection. These observations impact our view of eukaryotic diversity and offer future challenges for cell biology.

Comparison between Apicystis cryptica sp. n. and Apicystis bombi (Arthrogregarida, Apicomplexa): Gregarine parasites that cause fat body hypertrophism in bees.

The molecular divergence, morphology and pathology of a cryptic gregarine that is related to the bee parasite Apicystis bombi Lipa and Triggiani, 1996 is described. The 18S ribosomal DNA gene sequence of the new gregarine was equally dissimilar to that of A. bombi and the closest related genus Mattesia Naville, 1930, although phylogenetic analysis supported a closer relation to A. bombi. Pronounced divergence with A. bombi was found in the ITS1 sequence (69.6% similarity) and seven protein-coding genes (nucleotide 78.05% and protein 90.2% similarity). The new gregarine was isolated from a Bombus pascuorum Scopoli, 1763 female and caused heavy hypertrophism of the fat body tissue in its host. In addition, infected cells of the hypopharyngeal gland tissue, an important excretory organ of the host, were observed. Mature oocysts were navicular in shape and contained four sporozoites, similar to A. bombi oocysts. Given these characteristics, we proposed the name Apicystis cryptica sp. n. Detections so far indicated that distribution and host species occupation of Apicystis spp. overlap at least in Europe, and that historical detections could not discriminate between them. Specific molecular assays were developed that can be implemented in future pathogen screens that aim to discriminate Apicystis spp. in bees.

Molecular characterisation and morphological description of two new species of Hepatozoon Miller, 1908 (Apicomplexa: Adeleorina: Hepatozoidae) infecting leukocytes of African leopards Panthera pardus pardus (L.).

BACKGROUND: The African leopard Panthera pardus pardus (L.) is currently listed as a vulnerable species on the IUCN (International Union for the Conservation of Nature) red list of threatened species due to ongoing population declines. This implies that leopard-specific parasites are also vulnerable to extinction. Intracellular apicomplexan haemoparasites from the genus Hepatozoon Miller, 1908 have been widely reported from wild carnivores in Africa, including non-specific reports from leopards. This paper describes two new haemogregarines in captive and wild leopards from South Africa and provides a tabular summary of these species in relation to species of Hepatozoon reported from mammalian carnivores. METHODS: Blood was collected from nine captive and eight wild leopards at various localities throughout South Africa. Thin blood smears were Giemsa-stained and screened for intraleukocytic haemoparasites. Gamont stages were micrographed and morphometrically compared with existing literature pertaining to infections in felid hosts. Haemogregarine specific primer set 4558F and 2733R was used to target the 18S rRNA gene for molecular analysis. Resulting sequences were compared to each other and with other available representative mammalian carnivore Hepatozoon sequences from GenBank. RESULTS: Two species of Hepatozoon were found in captive and wild leopards. Of the 17 leopards screened, eight were infected with one or both morphologically and genetically distinct haemogregarines. When compared with other species of Hepatozoon reported from felids, the two species from this study were morphometrically and molecularly distinct. Species of Hepatozoon from this study were observed to exclusively parasitize a particular type of leukocyte, with Hepatozoon luiperdjie n. sp. infecting neutrophils and Hepatozoon ingwe n. sp. infecting lymphocytes. Phylogenetic analysis showed that these haemogregarines are genetically distinct, with Hepatozoon luiperdjie n. sp. and Hepatozoon ingwe n. sp. falling in well supported separate clades. CONCLUSIONS: To our knowledge, this is the first morphometric and molecular description of Hepatozoon in captive and wild African leopards in South Africa. This study highlights the value of using both morphometric and molecular characteristics when describing species of Hepatozoon from felid hosts.

Formin' an invasion machine: actin polymerization in invading apicomplexans.

Apicomplexan parasites are motile and invade host cells. The force required for this is generated by an actomyosin motor. In a recent paper, Baum and colleagues suggest that the protein formin regulates the polymerization of actin at the moving junction between parasite and host cell. This finding provides novel insight into the mechanism of host cell invasion.

Building the perfect parasite: cell division in apicomplexa.

Apicomplexans are pathogens responsible for malaria, toxoplasmosis, and crytposporidiosis in humans, and a wide range of livestock diseases. These unicellular eukaryotes are stealthy invaders, sheltering from the immune response in the cells of their hosts, while at the same time tapping into these cells as source of nutrients. The complexity and beauty of the structures formed during their intracellular development have made apicomplexans the darling of electron microscopists. Dramatic technological progress over the last decade has transformed apicomplexans into respectable genetic model organisms. Extensive genomic resources are now available for many apicomplexan species. At the same time, parasite transfection has enabled researchers to test the function of specific genes through reverse and forward genetic approaches with increasing sophistication. Transfection also introduced the use of fluorescent reporters, opening the field to dynamic real time microscopic observation. Parasite cell biologists have used these tools to take a fresh look at a classic problem: how do apicomplexans build the perfect invasion machine, the zoite, and how is this process fine-tuned to fit the specific niche of each pathogen in this ancient and very diverse group? This work has unearthed a treasure trove of novel structures and mechanisms that are the focus of this review.

The glideosome: a molecular machine powering motility and host-cell invasion by Apicomplexa.

The apicomplexans are obligate intracellular protozoan parasites that rely on gliding motility for their migration across biological barriers and for host-cell invasion and egress. This unusual form of substrate-dependent motility is powered by the "glideosome", a macromolecular complex consisting of adhesive proteins that are released apically and translocated to the posterior pole of the parasite by the action of an actomyosin system anchored in the inner membrane complex of the parasite. Recent studies have revealed new insights into the composition and biogenesis of Toxoplasma gondii myosin-A motor complex and have identified an exciting set of small molecules that can interfere with different aspects of glideosome function.