The Life Cycle of Rotaliella elatiana N. Sp.: A Tiny Macroalgavorous Foraminifer from the Gulf of Elat1

ABSTRACT. A new foraminiferan species, Rotaliella elatiana n. sp., was isolated in the Gulf of Elat, where it lives in association with a macrophytic green alga, Enteromorpha. The agamont of this tiny new species has a transparent test composed of a bilocular embryonic chamber followed by six to seven trochospirally coiled inflated chambers. The spiral sutures are undulated. The umbilical side has numerous denticules and has radial grooves. The gamont has only one inflated chamber. Rotaliella elatiana has a classical, heterophasic life cycle, with a regular alternation of diploid agamontic phase and haploid gamontic phase. The gamontic phase of the life cycle is exceptionally reduced and the uninucleated gamonts pair immediately after they build their first chamber. A few cases of autogamic reproduction were observed. R. elatiana is a heterocaryotic species; agamonts have one somatic and two to three generative nuclei.     

Ultrastructural Features of the Apical Complex,Pellicle, and Membranes Investing the Gamonts of Haemogregarina magna (Apicomplexa: Adeleina)1

ABSTRACT. Mature gamonts of Haemogregarina magna lie within a type of parasitophorous vacuole (Pv) apparently unique to the haemogregarines. The cytoplasm of infected erythrocytes was separated from the parasite by two Pv membranes. An additional membrane, coated on both sides with electron-dense material, closely invested the gamonts. The apical complex of the gamonts includes a conoid, two preconoidal rings, and an elaborate polar ring complex. The latter consisted of the polar ring and approximately 78 posteriorly directed, radially arranged, “tine-like” structures which fuse as they merge anteriorly into the polar ring. Freeze fracture replicas demonstrated that the pellicle of gamonts of H. magna was structurally similar to that of other apicomplexans. The closely apposed inner membranes of the pellicle formed plates which were arranged into strips along the long axis of the gamont. Calculations indicated that 13 such strips are found around the circumference of the gamonts with about six subpellicular microtubules associated with the inner surface of each strip. Gamonts of H. magna share many structural similarities with the kinetes, ookinetes, and sporokinetes of other apicomplexans. We propose that the conoid and polar ring complex are fundamental features of all apicomplexan “kinetes.”     

Fine Structure of Hepatozoon mocassini (Apicomplexa,Eucoccidiorida) Gamonts and Modifications of the Infected Erythrocyte Plasmalemma1

Transmission electron microscopy and scanning electron microscopy were used to investigate the fine structure of Hepatozoon mocassini gamonts and modifications of the infected erythrocyte plasmalemma. Intraerythrocytic gamonts were contained within a parasitophorous vacuole. An electron-lucid space observed between the gamont pellicle and the membrane of the vacuole corresponded to the unstained space described in light microscopy studies. Gamonts possessed a conoid, polar ring, subpellicular microtubules, four pairs of rhoptries, micronemes, ovoid granules, mitochondria with tubular cristae, and a pellicle composed of three individual unit membranes. The conoid had an anterior diameter of 320 nm, a posterior diameter of 360 nm, and a length of 150 nm. In contrast to a report on Hepatozoon aegypti, no micropore or “canopy-like structure” was observed. The plasmalemma of infected erythrocytes exhibited two types of modifications: gross membrane deformations and knobs with an electron-dense central mass. These knobs are structurally distinct from previously described membrane excrescences.     

Ultrastructural features of the apical complex, pellicle, and membranes investing the gamonts of Haemogregarina magna (Apicomplexa: Adeleina)

Mature gamonts of Haemogregarina magna lie within a type of parasitophorous vacuole (Pv) apparently unique to the haemogregarines. The cytoplasm of infected erythrocytes was separated from the parasite by two Pv membranes. An additional membrane, coated on both sides with electron-dense material, closely invested the gamonts. The apical complex of the gamonts includes a conoid, two preconoidal rings, and an elaborate polar ring complex. The latter consisted of the polar ring and approximately 78 posteriorly directed, radially arranged, "tine-like" structures which fuse as they merge anteriorly into the polar ring. Freeze fracture replicas demonstrated that the pellicle of gamonts of H. magna was structurally similar to that of other apicomplexans. The closely apposed inner membranes of the pellicle formed plates which were arranged into strips along the long axis of the gamont. Calculations indicated that 13 such strips are found around the circumference of the gamonts with about six subpellicular microtubules associated with the inner surface of each strip. Gamonts of H. magna share many structural similarities with the kinetes, ookinetes, and sporokinetes of other apicomplexans. We propose that the conoid and polar ring complex are fundamental features of all apicomplexan "kinetes."     

Septate Gregarines From Reticulitermes Flavipes and Reticulitermes Virginicus (Isoptera: Rhinotermitidae)

ABSTRACT. The gregarine parasites of Reticulitermes virginicus and Reticulitermes flavipes begin their development as trophozoites attached to the midgut epithelium by a small button-shaped epimerite. the epimerite is lost when the parasite becomes free-living in the gut lumen as a solitary gamont. Syzygy is late and was not observed. When full-grown, gamonts enter the hemocoel and fuse in pairs to form large gametocysts that are attached to the midgut of the termite by a duct. Thousands of sporocysts are formed within the original gametocyst. the mature sporocysts are released into the lumen of the midgut through the connecting duct. They are then passed out with the feces. These gregarines are believed to be identical to Gregarina termitis Leidy which was described from a single gamont and later erroneously placed in the genus Hirmocystis by Henry.     

Antigenic analysis of gamonts of Hepatozoon canis purified from leukocytes

Hepatozoon canis is a tick-borne apicomplexan parasite of dogs that infects neutrophils and parenchymal tissues. To study the antigenic characteristics of this parasite, a technique was devised for the purification of gamonts from peripheral blood neutrophils. White blood cells were separated on Ficoll-Hypaque density gradients and the gamonts were released from the host neutrophils by nitrogen cavitation. The blood used for purification originated from dogs with natural or experimental infections of H. canis with a parasitemia of 1.4-33%. The number of parasites collected ranged from 1.5 X 10(6) to 4.2 X 10(7). Portions of purified gamonts were separated and examined under phase and scanning electron microscopy, and the remaining purified parasites were then used as a source of antigens to characterize the humoral immune response by western blot analysis. Serum antibodies from infected dogs recognized more than 15 gamont antigens, and the antigenic patterns observed with sera from naturally and experimentally infected dogs were nearly similar. Four immunodominant protein bands of relative molecular weights of 107, 88, 63, and 28 kDa were recognized by all of the sera examined. The technique applied here for the isolation of host cell-free gamonts will facilitate studies on antigenic composition and immune responses against H. canis and on antigenic relationships between Hepatozoon from different host species and geographic regions.     

Observation by electron microscopy of a gregarine parasite of Antarctic krill: its histological aspects and ecological explanations

Antarctic krill are parasitized by gregarines (Phylum Apicomplexa, Class Sporozoea, Order Eugregarinida), which were observed in this study by light, scanning, and transmission electron microscopy. Eighty seven percentage of the krill examined (n = 93) were infected with the parasites in the cephalin stage, which were regularly found in the hind-gut epithelium of their host. Cephalins were found attached to the epithelium, and then were liberated into the intestinal lumen. The gamont stage, which follows the cephalin stage, was found in the intestinal lumen, as well as the diverticulum of mid-gut gland. However, gamonts found in the mid-gut gland were considerably larger and elongated in comparison with those in the intestinal lumen. Gamonts in the diverticulum appear to damage microvilli, which are involved in the uptake of digested nutrients and secretion of various enzymes. Therefore, elevated infestation of this parasite in the mid-gut gland may have a significant impact on the nutritional state of the Antarctic krill host.     

Host-parasite relationships of Ascogregarina chagasi (Eugregarinorida, Aseptatorina, Lecudinidae) in Lutzomyia longipalpis (Diptera: Psychodidae)

The life cycle of Ascogregarina chagasi in larvae and in adult female Lutzomyia longipalpis was studied by light and electron microscopy. Sporozoites and young gamonts were attached to the epithelial lining of the larval midgut via an osmiophilic contact zone. The mucron of young gamonts was bordered by an invaginated pellicular fold, and an electron-opaque vesicular structure was observed adjacent to it. Sporozoites possessed an apical complex and were bound by a double membrane with underlying subpellicular microtubules. The gamont pellicle was uniformly corrugated and consisted of two cortical membranes and a plasma membrane. Mature gamonts and gametocysts were found in the posterior ectoperitrophic space of third and fourth instar larval midguts and in the haemocoel of adult flies. Gametocysts in adult females adhered to the genital accessory glands, where they were encased in electron-dense capsules secreted by the fly through haemocyte-mediated humoral immune reactions. Oocytes were spindle-shaped and bound by a double-layered wall with a discernible polar plug at each end. Sporulation was endogenous, occurring within gametocysts in the midguts of larvae or the accessory glands of adult females. FITC-phalloidine staining of all stages of A. chagasi except mature gametocysts produced bright fluorescence, indicating the presence of a diffuse, actin-like protein in the cytoplasm.     

Gregarina coronata N. Sp. (Apicomplexa: Eugregarinida) Described from Adults of the Southern Corn Rootworm, Diabrotica undecimpunctata howardi (Coleoptera: Chrysomelidae)

ABSTRACT. Gregarina coronata n. sp. (Apicomplexa: Eugregarinida) is described from the adults of the Southern Corn Root Worm, Diabrotica undecimpunctata howardi (Coleoptera: Chrysomelidae). Measurements given are means, in micrometers, taken from mature gamonts in association. Primite: protomerite hemi-ellipsoidal with basal tumidus, length 47.6, width 44.0, with cytoplasmic granule, apical crown apparent; deutomerite elongate ellipsoidal, length 227.9, width 81.3; epimerite absorbed into anterior in gamont, globular in trophozoite. Satellite: protomerite hemi-ellipsoidal, truncated at association interface to appear trapezoidal, length 39.2, width 49.6, with cytoplasmic granule; deutomerite elongate ellipsoidal, length 240.6, width 80.2; epimerite absorbed into anterior in gamont. Association caudofrontal and often precocious, occurring during growth of trophozoites. Gametocysts spherical, 115.3 in diameter, 132.9 with hyaline coat; producing multiple oocyst chains under moist storage in 24–36 h. Oocysts very uniform in shape and size, dorsad: doliform, length 6.4, equatorial width 3.4, polar width 2.9; pleuron: dorso-ventrally flattened, corpus concave with bicondylic termina; corpus height 0.98, width 4.44; terminus height 1.96, width 0.98.     

Etude de l'Ultrastructure de Gregarina polymorpha (Hamm.) en Syzygie

SYNOPSIS. The electron-microscope study of syzygy in Gregarina polymorpha , shows that this coupling, a prelude to cyst formation and to sexual phenomena, must not be considered as a simple pairing of individuals, but is rather the response of a gamont to a stimulus coming from another individual of a different sex. This response, in the species studied, can be recognized from the modifications that will appear progressively at the contact surfaces of the two gregarines. They affect only the protomerite apex of the posterior individual. There occur: rapid growth of the protomeritic epicyte; separation of this epicyte from the fibrillar zone underneath; formation of a protomeritic cup containing various inclusions; the disappearance of cuticular digitations and the formation of invaginations which become deeper and deeper.
All these modifications occur as soon as syzygy starts. They seem to indicate a reaction of the satellite epicyte to a possible stimulus coming from the anterior female gamont. This epicyte could play a different part from the initial one.
In the case of association between a primite and several satellites, the posterior gamonts react in a comparable manner but not together.     

Uniform Terminology for the Protozoan Subphylum Apicomplexa*

SYNOPSIS. Uniform names for the stages, processes and structures of apicomplexan protozoa are proposed and defined, and names that should be superseded are listed. The same names are used for the same stages of all members of the group. Gregarines are designated as either septate or aseptate rather than as polycystid or monocystid. The gregarine stage often called a sporadin is recognized for what it is, a gamont. The cyst formed around 2 gregarine gamonts in which zygotes are formed is a gametocyst; it contains oocysts which in turn contain sporozoites. The term “spore” is inappropriate for these oocysts. The apical complex includes the polar ring, conoid, rhoptries, micronemes and subpellicular tubules. The gregarine “pseudocyst” is actually a gametocyst residuum. The term micropore is preferred to cytostome for the apicomplexan structure, since it is visible only with the electron microscope.     

Susceptibility of Aedes aegypti and Aedes albopictus larvae to Ascogregarina culicis and Ascogregarina taiwanensis (Apicomplexa: Lecudinidae) from Florida

The susceptibility of Aedes aegypti to Ascogregarina culicis and Aedes albopictus to Ascogregarina taiwanensis was examined with mosquito and parasite strains from Tampa, FL. When each host was bioassayed with its natural gregarine, the infection intensity indicated that Ae. aegypti was 59% more susceptible to A. culicis (87 gamonts/larva) than Ae. albopictus to A. taiwanensis (47 gamonts/larva). Infections in single and mixed host populations exposed to 100 oocysts/larva of one and both parasites demonstrated that Ae. aegypti harbors higher A. culicis gamont loads than Ae. albopictus of A. taiwanensis. In dual gregarine exposures of single host populations, the A. culicis infection intensity in Ae. aegypti was reduced by approximately 50%. A. taiwanensis exhibited the same capability of infecting Ae. albopictus in single and dual exposures. In mixed host populations there were no cross infections, but A. taiwanensis in Ae. albopictus produced an infection intensity of approximately 70% lower than that of A. culicis in Ae. aegypti.     

The Fine Structure of Some Developmental Stages of Mattesia grandis McLaughlin (Sporozoa, Neogregarinida), a Parasite of the Boll Weevil Anthonomus grandis Boheman

SYNOPSIS Sporozoites, macronuclear schizonts, merozoites and gamonts of Mattesia grandis were examined by electron microscopy. A conoidal complex, consisting of conoid, polar rings and subpellicular microtubules was present in all of these stages. The conoidal complex was similar in structure to the same organelle of other Sporozoa. The conoidal complex in mono- to quadrinucleate macronuclear schizonts is transformed into an organelle similar to the mucron of some eugregarines.
This mucron consists of a specialized area of the cell membrane from which fine fibers extend into a large vacuole situated directly beneath the cell membrane. The top part of the vacuole is encircled by 2 ring-like structures formed by the dilatation of the original apical rings. The vacuole of the mucron contains many anastomosing protrusions of the cytoplasm, suggesting a nutritional role. The mucron disappears when the schizont reaches the multinucleate state. Later the merozoites bud from the surface of the schizont as in the coccidia. Each merozoite again has a conoidal complex, which persists thru the gamont stage and usually serves as the point of contact between 2 gamonts during their pairing.
The presence of a conoidal complex thru a major portion of the life cycle, its transformation into a mucron and the mode of formation of merozoites indicate that the Neogregarinida combine the fine structure characters of both the Eugregarinida and the Eucoccida, thereby suggesting a phylogenetic relationship between these sporozoans, with the neogregarines as a link between eugregarines and coccidia.     

A new species of <Emphasis Type="Italic">Hepatozoon</Emphasis> Miller, 1908 (Apicomplexa: Adelerina) from the snake <Emphasis Type="Italic">Philodryas nattereri</Emphasis> Steindachner (Squamata: Dipsadidae) in northeastern Brazil

Based on both unique morphological characteristics of the gamont, distinct changes caused to the host erythrocyte and analysis of partial 18S rRNA gene sequences, a new parasite of the genus Hepatozoon Miller, 1908 is described from the snake Philodryas nattereri Steindachner (Squamata: Dipsadidae) in northeastern Brazil. The new species, Hepatozoon musa n. sp., is characterized by large and curved mature gamonts (18.9 ± 0.9 μm in length and 3.8 ± 0.3 μm in width) that considerably engorge infected host erythrocytes and displace the nucleus laterally, which become longer and thinner. Phylogenetic estimates indicate the new species is more closely related to the recently described Hepatozoon cuestensis O’Dwyer, Moço, Paduan, Spenassatto, Silva & Ribolla, 2013, from Brazilian rattlesnakes. These recent findings highlight the need for further studies of Hepatozoon to better determine the biodiversity of this common but poorly-studied parasite group.     

Hepatozoon polytopis n. sp. parasitic in two genera and species of colubrid snakes in southern Florida

Hepatozoon polytopis, described from Coluber constrictor priapus from Palm Beach County, Florida, has short, usually broad gamonts 12.8 x 4.6 microm (10.0-15.0 x 3.5-6.0), with LW 58.5 microm2 (42-84) and L/W 2.84 (1.8-3.7). Nuclei commonly extend into first quarter of gamont (45%), are always present in second quarter, and seldom in third quarter (11%), with dimensions 4.5 x 3.4 (3.0-6.0 x 2.5-4.5) and LW 15.1 (10.0-24.0). Spherical to ovoid oocysts, 122.1 x 104.9 (62-240 x 57-190), with L/W 1.17 (1.0-1.9), contain 31.3 (3-103) sporocysts. Spherical to ovoid sporocysts, 38.0 x 33.9 (28-73 x 25-58), with LW 1,325.1 (756-4,168) and L/W 1.12 (1.0-1.4), contain 42.9 (22-64) sporozoites. Thamnophis sauritus sackenii from Palm Beach County is infected also by H. polytopis, as indicated from similar gamont dimensions and verified by isolation of an identical haplotype of the 18S ribosomal RNA gene from both host species.     

Dynamic organization of microtubules and microtubule-organizing centers during the sexual phase of a parasitic protozoan, Lecudina tuzetae (Gregarine, Apicomplexa)

Lecudina tuzetae is a parasitic protozoan (Gregarine, Apicomplexa) living in the intestine of a marine polychaete annelid, Nereis diversicolor. Using electron and fluorescence microscopy, we have characterized the dynamic changes in microtubule organization during the sexual phase of the life cycle. The gametocyst excreted from the host worm into seawater consists of two (one male and one female) gamonts in which cortical microtubule arrays are discernible. Each gamont undergoes multiple nuclear divisions without cytokinesis, resulting in the formation of large multinucleate haploid cells. After cellularization, approximately 1000 individual gametes are produced from each gamont within 24 h. Female gametes are spherical and contain interphase cytoplasmic microtubule arrays emanating from a gamma-tubulin-containing site. In male gametes, both interphase microtubules and a flagellum with "6 + 0" axonemal microtubules extend from the same microtubule-organizing site. At the beginning of spore formation, each zygote secretes a wall to form a sporocyst. Following meiotic and mitotic divisions, each sporocyst gives rise to eight haploid cells that ultimately differentiate into sporozoites. The ovoid shaped sporocyst is asymmetric and forms at least two distinctive microtubule arrays: spindle microtubules and microtubule bundles originating from the protruding apical end corresponding to the dehiscence pole of the sporocyst. Because antibodies raised against mammalian centrosome components, such as gamma-tubulin, pericentrin, Cep135, and mitosis-specific phosphoproteins, react strongly with the microtubule-nucleating sites of Lecudina, this protozoan is likely to share common centrosomal antigens with higher eukaryotes.     

Light optical and electron microscopic study of the cystic stages in Sarcocystis fusiformis from the buffalo

Three different cell types are distinguished in the sarcocysts of Sarcocystis fusiformis from the water buffalo: metrocytes, intermediate cells, and merozoites. The former lines the cyst near the border, the two latter lie more inside forming groups of cells separated from each other by septae. The pattern of metrocyte division giving rise to another metrocyte population has not been observed, still remaining obscure. Merozoites do not divide asexually due to their gamont nature to be realized in the final host only. The intermediate cells divide asexually by endodyogeny giving rise, on the one hand, to another population of intermediate cells, and on the other--to merozoites which divide no longer. Cytophotometrical measurements (G. D. Gaibova, 1987) revealed the amounts of DNA per cell nucleus within 1 and 2 c. This quantity corresponds to the stable DNA content in the most numerous population of merozoites (gamonts), whereas the amount between those higher than 1 c and 2 c may be attributed to the nuclei of metrocytes and intermediate cells, respectively.     

Ultrastructural Observations on the Gametocytic Stages of the Coccidium Tyzzeria chalcides Probert, Roberts & Wilson, 1988 from the Ocellated Skink Chalcides ocellatus

Gametogenesis of Tyzzeria chalcides Probert, Roberts & Wilson, 1988, from the ocellated skink, Chalcides ocellatus , occurs within the epithelium of the gali bladder. Transmission electron microscopy reveals that macrogamonts contain 2 types of wall-forming bodies. Type I bodies are large densely stained structures associated with rough endoplasmic reticulum and the Golgi apparatus. They appear to be formed within the Golgi itself. Type II bodies are less densely stained, smaller and appear to form directly from the rough endoplasmic reticulum. Canaliculi are associated with Type I wall-forming bodies and probably function to transport the wall-forming bodies to the pellicle. Micropores occur in the pellicie and large amylopectin granules, lipid globules and dense bodies are found within the cytoplasm of the macrogamont. Mature microgamonts contain in excess of 20 microgametes, each of which has 2 flagella and an associated mitochondrion. Both types of gamont are found within a parasitophorous vacuole, in the host cell, which is filled with vesicular material on which the gamonts probably feed.     

Sexual Reproduction of the Foraminiferan Planoglabratella opercularis (d'Orbigny, 1839) in Nature

The taxonomic composition of the entire foraminiferan assemblage in the southern part of Peter the Great Bay (Sea of Japan) was studied during the summer 1999 expedition. Along with other foraminiferans, several offspring-filled tests of Planoglabratella opercularis (d'Orbigny, 1839) were found in surface sediment samples. Based on the extensive literature on the reproductive cycle of foraminiferans and indirect evidence, it was found that each of the parent individuals of this species was a partner in a plastogamous pair, thus being a gamont, while their offspring were agamonts. The gamogony stage, contrary to the generally accepted opinion, took place under natural conditions in summer (not in fall or winter) at a bottom water temperature of 15–18°C. The results of this research may be important in paleogeographical and paleoecological reconstructions.