Fine Structure of the Oocyst Wall of Eimeria nieschulzi

SYNOPSIS. Oocysts of Eimeria nieschulzi from the laboratory rat, Rattus, norvegicus , were studied by scanning and transmission electron microscopy. Oocysts had a rough outer wall with apparent random depressions. The oocyst wall is composed of 2 layers: an osmiophilic outer layer consisting of a rough external and smooth internal surface, and a relatively thick, electron-lucent inner layer. The outer layer is composed of a dense, coarsely granular matrix. The inner layer consists of homogeneous fine granular material interspersed with coarse osmiophilic granules and contains one closely applied membrane on the outermost surface. Several raised lenticular areas are seen on the coarse outer surface of the inner layer. These layers are 102 (75–128) and 176 (135–204) nm thick, respectively.
The sporocyst wall is thin, consisting of 3 to 4 unit membranes, and measures 27 (18–34) nm thick.     

Nematopsis gigas n. sp. (Apicomplexa), a parasite of Nerita ascencionis (Gastropoda, Neritidae) from Brazil

A new species of Nematopsis (Apicomplexa, Porosporidae) is described from the mantle tissues of the seawater gastropod, Nerita ascencionis (Neritidae), collected in the Atlantic North off the coast of "Fernando de Noronha" Island (3 degrees 47' 57' S, 32 degrees 25' 12' W) situated about 350 km from the northeast coast of Brazil. Numerous oocysts, each contained in a parasitophorous vacuole, were found in the cytoplasm of phagocytes in the mantle tissue of the host. The phagocytes were surrounded by a thin wall composed of lucent material. The phagocyte cytoplasm contained a nucleus surrounded by numerous vesicles and some dense masses. The oocysts were 21.9 +/- 0.5 microm long, and 11.5 +/- 0.6 microm wide. The oocyst wall was 0.18-0.25 microm thick, and the apical zone contained a micropyle, 1.0-1.2 microm in diameter, covered by a canopy-like operculum about 0.25 microm thick. Externally, the oocyst wall was surrounded by numerous anastomosing microfibrils attached to the wall and extending towards the periphery of the parasitophorous vacuole. Some microfibrils formed a dense complex network that surrounded the oocyst in the middle of the parasitophorous vacuole, which opened only at the apical zone near the external region of the opercular system. On the basis of the data obtained by light and transmission electron microscopy and host specificity, the gregarine Nematopsis gigas is distinguished from the nearest species as a new species. The taxonomic affinities and morphological comparisons with other similar species of the same genus are discussed.     

A new and unusual eimerian (protozoa: Eimeriidae) from the liver of the Gulf killifish, Fundulus grandis

Oocysts and sporocysts of Eimeria funduli sp. n. are described from the Gulf killifish, Fundulus grandis, on the basis of light microscopy, transmission and scanning electron miscroscopy, and location in the liver of infected hosts. The spherical sporulated oocysts of E. funduli isolated from liver tissue measure 20-31 (25) micrometer across with ovoid sporocysts 9-11 X 5-7 (10 X 6) micrometer. A micropyle, polar granule, and oocyst residuum are absent, but sporocysts have Stieda and substieda bodies, a few residual granules, and 10-25 (15) unique projecting structures with expanded distal portions that we term "sporopodia". Sporopodia 1-3 (2) micrometer high support a transparent membrane that completely surrounds the sporocyst. Sporozoites have one large posterior refractile body. Ultrastructurally, the oocyst wall consists of two thin layers of granular material: an electron-dense outer layer with a rough external surface and an electron-lucent inner one of approximately equal thickness. One or two unit membranes line the inner surface of the inner layer. Each layer is 40-60 (55) nm thick. The sporocyst wall, 78-130 (110) nm thick, consists of an electron-lucent material with the outer surface being more electron dense and giving rise to osmiophilic sporopodia; closely associated with these and the outer surface are one or two unit membranes. A thin osmiophilic layer of fine granular material lines the inner surface.     

Two new species of coccidia (apicomplexa: Eimeriidae) from the bearded false chameleon Chamaeleolis barbatus (Sauria: polychridae) from cinco pesos, Pinar Del Río, Cuba.

Parasitological examination of bearded false chameleons Chamaeleolis barbatus freshly imported from Cuba revealed the presence of 2 species of coccidia that are described as new. Oocysts of Isospora chamaeleolidis n. sp. are spherical to slightly subspherical, 16.1 (13-21) x 15.6 (13-19) microm, with a brownish and bilayered wall approximately 1.0-1.5 microm thick; outer layer markedly pitted. 0.75-1.0 microm thick. One, rarely 2, globular polar granules, 1.5 in diameter are present in the sporulated oocysts. Sporocysts are ellipsoidal, 10.8 (10-13) x 7.8 (7-9) microm, with a smooth, colorless, and unilayered sporocyst wall. Stieda body and substieda bodies are present. A sporocyst residuum is present, consisting of small granules of irregular size scattered among the sporozoites. Oocysts of Eimeria chamaeleolidisbarbati n. sp. are broadly oval, 19.0 (17-21) x 15.7 (15-17) microm, with a bilayered, colorless oocyst wall approximately 0.75 thick; outer layer of oocyst wall is smooth, 0.5 microm thick. One or 2, rarely 4, globular, irregular polar granules, approximately 1.5 microm in diameter, are present in sporulated oocysts. Sporocysts are broadly oval, 7.4 (7-8.5) x 6.1 (5.5-7) microm, with a smooth, colorless, and unilayered sporocyst wall, composed of 2 valves joined by suture; Stieda body and substieda bodies are absent.     

Goussia girellae n. sp. (Apicomplexa: Eimeriorina) in the opaleye, Girella nigricans

Goussia girellae n. sp. is described from the opaleye fish, Girella nigricans. Merogonic stages were observed in the apices of intestinal epithelial cells, in the lamina propria, and in extra-intestinal sites including liver, gills, and spleen. Gamonts were observed in the intestinal epithelial cells. Only unsporulated oocysts were detected in the intestine, and sporulation occurred when feces containing oocysts were incubated for 48 h in seawater at 21 degrees C. Oocysts are elongated (24.8 x 14.7 micron) with a wall about 200 nm thick and have no residuum, micropyle, or polar granule. Sporocysts are ellipsoid (8.5 x 4.5 micron), have a thin two-layered wall approximately 30 nm thick, and consist of two valves joined by a suture. Although moribund opaleye were also infected with Gyrodactylus sp., Cryptobia sp., Cardicola sp., and epitheliocystis organisms (chlamydia), all fish were heavily infected with G. girellae and morbidity was thus attributed to the coccidium.     

Eimeria crotalviridis sp. n. from Prairie Rattlesnakes, Crotalus viridis viridis, in New Mexico with Data on Excystation of Sporozoites and Ultrastructure of the Oocyst Wall

SYNOPSIS Oocysts of Eimeria crotalviridis sp. n. are described from prairie rattlesnakes, Crotalus viridis viridis in New Mexico on the basis of light and electron microscopy and in vitro excystation of sporozoites. Sporulated oocysts of E. crotalviridis are elliptical, 26.4 × 22.3 (23–29 × 20–24) μm with ovoid sporocysts 11.7 × 8.1 (11–13 × 7–9) μm. A micropyle, micropyle cap and polar bodies are absent, but oocyst and sporocyst residua and Stieda and substieda bodies are present. Excysted sporozoites are 12.4 × 2.8 (11–13 × 2–3) μm and have 1 large posterior refractile body and a nucleus with a prominent nucleolus. Ultrastructurally, the oocyst wall has 2 layers, a thick, electron-dense, highly sculptured outer layer composed of a fine granular matrix and a thin, granular, osmiophilic inner layer, separated from the outer layer by at least one unit membrane. These layers are 441 (353–510) and 21.6 (19–29) nm thick, respectively. Within 15 min after exposure to a trypsin-sodium taurocholate fluid, sporozoites of E. crotalviridis excysted from 5-month-old sporocysts.     

Ultrastructural aspects of hepatic coccidiosis caused by Goussia lusca n. sp. (Apicomplexa: Coccidia) infecting Trisopterus luscus (Gadidae) from the NE Atlantic Ocean

Goussia lusca n. sp. is described from the liver of pouting Trisopterus luscus from the NE Atlantic Ocean in Ibero-Atlantic Portuguese and Spanish waters. Mature oocysts were 31.7 (28.8 to 35.4) microm in diameter. Each oocyst contained 4 ellipsoidal sporocysts arranged in an aleatory position, and measuring approximately 13.7 x 9.2 microm. Each sporocyst contained 2 sporozoites. Ultrastructurally, the sporocyst wall consisted of a dense inner layer 115 nm thick, transversely striated, regularly intercalated by thin grooves with electron-lucent spaces, and separated from the outer layer by a thin, light (electron-lucent) space. The outer layer was multilamellated and consisted of parallel dense bands alternating with light spaces. These lamellae formed filamentous extensions of the wall. The dehiscence suture, a characteristic feature of the genus, was present in the sporocysts. No external clinical signs were observed in the host fish. Parasites observed in the liver tissue were often enveloped in a yellowish-brown matrix, generally known as 'yellow bodies'. Sometimes sporocysts were observed in direct contact with the liver cells. Parasites in degeneration and aggregations of amylopectin granules were frequently observed surrounded by host inflammatory cells. In severe infections, we observed large agglomerations of oocysts encapsulated by layers of concentrically arranged connective tissue forming large granulomas, which caused significant replacement of the host liver parenchyma by the parasite.     

Fine Structure of Zygotes and Oocysts of Eimeria nieschulzi*

SYNOPSIS. Mature macrogamonts were present in the small intestine of rats 5.5 to 7.5 days postinoculation with Eimeria nieschulzi oocysts; oocysts were present at 6 to 7.5 days. Types I and II wall-forming bodies in macrogamonts began to undergo ultrastructural changes within zygotes to form the outer and inner layers of the oocyst wall. Before and during oocyst wall formation a total of 5 membranes (M1–5) were formed at or near the surface of the zygote. The outer and inner oocyst wall layers formed between M2 and M3, and M4 and M5, respectively. The mature oocyst was loosely surrounded by M1 and M2, had an electron-dense outer layer, 100–275 nm thick, and an electron-lucent inner layer, 160–180 nm thick. It also contained an electron-lucent line consisting of M3 and M4 interposed between the outer and inner layers of the oocyst wall. The micropyle, measuring 935 × 47 nm, was located in the outer layer of the oocyst wall and consisted of 10–14 alternating layers of electron-dense and lucent material. The sporont of mature oocysts was covered by M5, immediately beneath which were M6 and M7. The sporont contained a nucleus and nucleolus, lipid and amylopectin bodies, mitochondria, ribosomes, as well as smooth and rough endoplasmic reticulum. Canaliculi, Golgi complexes, and types I and II wall-forming bodies were absent.     

The fine structure of the developing oocyst of Pterospora floridiensis (Apicomplexa,Urosporidae)

Developing oocysts of the gregarine Pterospora floridiensis Landers 2001 were examined by transmission electron microscopy. Each oocyst had an outer capsule and an inner capsule that contained 8 sporozoites. In early stages of development the inner capsular wall was separated from the developing sporozoites and residual mass, and was not appressed to the sporozoites. Early stage sporozoites were connected to a residual mass and were filled with endoplasmic reticulum, golgi and numerous developing secretory vesicles. In late stages of oocyst and sporozoite development, the inner capsular wall was closely appressed to the sporozoite surface. The inner capsular wall was ～60-100 nm thick and the outer capsular wall was ～160-320 nm thick. There were no extensions on the outer wall for which the genus was named. Late stage sporozoites had no residual mass connection, were more electron dense, and contained three distinct types of dense secretory structures: 1) small oval/spherical dense vesicles, 2) large (350-400 nm) vesicles near the anterior end, and 3) elongated dense tubular bodies that converged at the apex. Few ultrastructural reports exist of developing gregarine oocysts and sporozoites, and as more studies are completed these morphological characteristics may be important in interpreting molecular phylogenetic analyses.     

Ultrastructural organization of Alicyclobacillus tolerans strain K1T cells

Electron microscopy examinations of thin sections and freeze-fracture replicas revealed the specific ultrastructural features of Alicyclobacillus tolerans strain K1(T). In particular, the cell wall displayed an ultrastructure typical of gram-positive bacteria and consisted of a thin murein layer (50-60 A in thickness); cells exhibited a surface S-layer constituted by large hexagonally packed (p6-symmetry) rod-shaped subunits of 150-160 A in diameter and 200 A in height. In the cytoplasmic membrane, there were intramembrane vesicular structures that sometimes appeared as large leaflets in the central part. The cytoplasm contained numerous vesicular inclusions covered with a monolayered wall, dissimilar to bilamellar lipid membranes. Endospore coats displayed an intricate structure and consisted of three thick layers; the outer layer had an unusual fine structure; the exosporium was also found.     

The structure and ultrastructure of the sinus venosus in the mature dogfish (Scyliorhinus canicula): the endocardium, the epicardium and the subepicardial space

The sinus venosus of fish is the most caudal chamber of the heart. It is often reduced in teleosts but well developed in elasmobranchs. The sinus venosus of the dogfish (Scyliorhinus canicula) is vital, since it harbours key elements such as a little known neuroendocrine system and the nodal tissue. However, the study of its structure is still incomplete. We examined the endocardium, epicardium and subepicardium of the sinus venosus in mature dogfishes. The wall is 100-250 microm thick and comprises three main layers. Large bundles of myocardial cells occupy the middle layer. The endothelial ensheathing is composed of thin endocardial cells with prominent nuclei towards the lumen, whose cytoplasm contains numerous dense bodies and moderately dense bodies, 150-800 nm in diameter and large vacuoles. The possible functions of these organelles are discussed. The outermost layer is made of a robust sheet of cuboidal epicardial cells separated from the subepicardium by a conspicuous basal lamina. Numerous microvilli towards the pericardial cavity and elliptical vesicles are located in the apex of epicardial cells. A thick layer richly endowed with dense bundles of collagen fibres forms the subepicardial space. This structure should be contrasted with the venous return mechanism of elasmobranchs.     

STUDIES ON THE OXIDATIVE METABOLISM OF SACCHAROMYCES CEREVISIAE : I. Observations on the Fine Structure of the Yeast Cell

Vegetative cells of Saccharomyces cerevisiae were fixed with potassium permanganate followed by uranyl nitrate, embedded in methacrylate, and studied in electron micrographs of thin sections. Details of the structure of the cell wall, cytoplasmic membrane, nucleus, vacuole, and mitochondria are described. Cell membranes, about 70 to 80 A thick, have been resolved into two dense layers, 20 to 25 A thick, separated by a light layer of the same dimensions, which correspond in thickness and appearance to the components of the "unit membrane" as described by Robertson (15). The cell wall is made up of zones of different electron opacity. Underlying the cell wall is the cytoplasmic membrane, a sinuous structure with numerous invaginations. The nucleoplasm, often of uneven electron opacity, is enclosed in a pair of unit membranes in which nuclear pores are apparent. The vacuole, limited by a single unit membrane, is usually irregular in outline and contains some dense material. Rod-shaped mitochondria, 0.4 to 0.6 µ in length and 0.2 to 0.3 µ in diameter, are smaller in size, but similar in structure to some of those described in plant and animal cells. Attempts to use osmium tetroxide as fixative were unsuccessful, a result similar to that obtained by other workers. It is suggested that yeast cells are impermeable to osmium tetroxide, except when grown under specific conditions.     

Goussia girellae N. Sp. (Apicomplexa: Eimeriorina) in the Opaleye,Girella nigricans1

ABSTRACT Goussia girellae n. sp. is described from the opaleye fish, Girella nigricans. Merogonic stages were observed in the apices of intestinal epithelial cells, in the lamina propria, and in extra-intestinal sites including liver, gills, and spleen. Gamonts were observed in the intestinal epithelial cells. Only unsporulated oocysts were detected in the intestine, and sporulation occurred when feces containing oocysts were incubated for 48 h in seawater at 21°C. Oocysts are elongated (24.8 × 14.7 μm) with a wall about 200 nm thick and have no residuum, micropyle, or polar granule. Sporocysts are ellipsoid (8.5 × 4.5 μm), have a thin two-layered wall approximately 30 nm thick, and consist of two valves joined by a suture. Although moribund opaleye were also infected with Gyrodactylus sp., Cryptobia sp., Cardicola sp., and epitheliocystis organisms (chlamydia), all fish were heavily infected with G. girellae and morbidity was thus attributed to the coccidium.     

Control of Morphogenesis in Geodermatophilus: Ultrastructural Studies

Geodermatophilus grows in two major forms, a nonmotile irregularly shaped aggregate of coccoid cells (C-form) and a motile budding rod (R-form). Morphogenesis can be controlled by an unidentified factor in Tryptose which is required for maintenance of the organism in the C-form and for differentiation of the R-form to the C-form. Morphogenetic events occur synchronously in the described system. Ultrastructural studies show that the major difference between C- and R-forms is in cells envelope structure. R-form cell walls consist of two layers, an inner transparent membranous layer (10 to 12.5 nm thick) and an outer dense diffuse layer (7.5 to 10 nm). In addition to these layers, the C-form has a thick fibrous layer (30 nm) over the dense layer. This layer appears to be a cementing substance which holds the coccoid cells together.     

Eimeria saudiensis n. sp. (Apicomplexa: Eimeriidae) from the Arabian oryx (Oryx leucoryx) in Saudi Arabia

Oocysts of Eimeria saudiensis n. sp. (Apicomplexa: Eimeriidae) are described from the feces of the Arabian oryx, Oryx leucoryx, from the Riyadh Zoo, Saudi Arabia. The oocysts were ellipsoidal or slightly ovoid, 31.2 x 24.5 (24.3-36.5 x 20.0-27.6) micron with a bilayered wall about 1.7 micron thick. The micropyle was covered by a dome-shaped cap. The oocyst residuum was absent, but tiny polar granules were present. The sporocysts were elongate ovoid, 14.3 x 7.2 (11.5-18.5 x 6.0-9.0) micron, had a Stieda body, but lacked a substiedal body. The sporocyst residuum was present, composed of numerous small granules. The sporozoites were elongate club-shaped, and contained two prominent refractile bodies.     

Vavraia lutzomyiae n. sp. (Phylum Microspora) infecting the sandfly Lutzomyia longipalpis (Psychodidae, Phlebotominae), a vector of human visceral leishmaniasis

Vavraia lutzomyiae (Microsporida; Pleistophoridae) is a new species parasitic in the tropical phlebotomine sandfly, Lutzomyia longipalpis (Diptera, Psychodidae, Phlebotominae), a major vector of Leishmania chagasi in Latin America where human visceral leishmaniasis is endemic. Infected larvae and pupae were parasitized in the abdomen, and some adults were parasitized in Malpighian tubules and midgut. The sporogonial plasmodium divided by multiple divisions into up to 64 uninucleate sporoblasts. These stages were surrounded outside the plasmalemma by a thick, amorphous dense coat and transformed into a merontogenetic sporophorous vesicle within which the sporonts developed into sporoblasts. The mature microsporidian spores were broadly ellipsoidal and measured 6.1+/-0.43 x 3.1+/-0.15 microm. The spore wall consisted of a transparent endospore (approximately 100 nm) and a thin electron dense exospore (approximately 30 nm) with the outer limit slightly undulated. Spores contained a polar filament arranged peripherally in a single layer of eight to nine wide anterior coils (approximately 125 nm diameter), and three to four narrow posterior coils (approximately 70 nm diameter). Transverse sections revealed a concentric layer organization with the internal layer surrounded by numerous (up to 25) longitudinal microfibrils. The angle of tilt of the polar filament was about 65-68 degrees.     

Caryospora uptoni n. sp. (Apicomplexa: Eimeriidae) from red-tailed hawks (Buteo jamaicensis borealis)

Oocysts of Caryospora uptoni n. sp. were described from the feces of red-tailed hawks, Buteo jamaicensis borealis. Sporulated oocysts were spherical or subspherical and measured 28.1 by 26.4 micron. The oocyst wall was composed of a yellowish outer layer and brownish inner layer and was about 1.5 micron thick. Neither micropyle, polar granules, nor oocyst residuum were present. A single, spherical sporocyst 18.2 by 17.9 micron was present; a Stieda body was absent. A spherical eccentrically located sporocyst residuum was present in many sporocysts, but it degenerated to form a dispersed granular residuum in other sporocysts. Eight randomly arranged sporozoites, 12.6 by 4.2 micron, were present in each sporocyst; they contained a centrally or slightly posteriorly located nucleus.     

Scanning and transmission electron microscopy of the oocyst wall of Isospora lacazei

The oocyst wall of Isospora lacazei from sparrows was studied with scanning (SEM) and transmission (TEM) electron microscopy. In TEM, the oocyst wall consisted of four distinct layers (L1-4). The innermost layer, L1, was moderately electron-lucent and 240--285 nm thick; L2 was electron-dense and 210--240 nm thick; L3 was moderately electron-lucent and 15--150 nm thick; L4, the outer most layer, was discontinuous and consisted of electron-dense discoid bodies which measured 180--220 nm x 320--840 nm. The discoid bodies of L4 as seen by TEM appeared spheroid in shape when observed by SEM. One or two membranes were situated on or between various layers of the oocyst wall. One such membrane occurred on the inner margin of L1, two closely applied membranes were interposed between L1 and L2, one membrane occurred between L2 and L3, and one membrane on the outer margin of L3.     

Coccidia of Brazilian mammals: Eimeria corticulata n. sp. (Apicomplexa: Eimeriidae) from the anteater Tamandua tetradactyla (Xenarthra: Myrmecophagidae) and Eimeria zygodontomyis n. sp. from the cane mouse Zygodontomys lasiurus (Rodentia: Cricetidae)

Feces from a specimen of Tamandua tetradactyla (Linn.) from Portel, Pará State, north Brazil, contained two different coccidial oocysts; one identified as Eimeria tamanduae Lainson 1968, and the other as a new species, described here as Eimeria corticulata n. sp. Oocysts of E. corticulata are ellipsoidal, 37.4 x 30.4 (31.2-43.7 x 23.7-35.0) microns, shape index (length/width) 1.2 (1.0-1.5). Oocyst wall 2.5-3.7 microns thick and composed of two layers; an outer thick, brown-yellow one with radial striations, and a thin inner smooth one: no visible micropyle. Oocyst residuum a large globule of about 10.7 x 10.3 microns, usually accompanied by a number of smaller attached globules. Sporocysts ellipsoidal, 21.0 x 11.0 (20.0-22.5 x 10.0-12.5) microns, with a conspicuous Stieda body; shape index 1.9 (1.6-2.2). Sporocyst residuum a small number of scattered granules: sporozoites 18.7 x 5.0 microns, with a large posterior refractile body. Eimeria zygodontomyis n. sp. is described in feces from Zygodontomys lasiurus (Lund) from the Serra dos Carajás, Pará. Oocysts ellipsoidal to cylindrical, 16.5 x 12.0 (13.7-18.7 x 11.2-12.3) microns, shape index 1.4 (1.2-1.5). Wall colorless, smooth, single-layered and about 0.6 micron thick: no micropyle. No oocyst residuum, but a polar granule of about 1.8 x 1.0 microns is sometimes present. Sporocysts ellipsoidal, 8.4 x 5.5 (7.5-8.7 x 5.0-6.2) microns, shape index 1.5 (1.4-1.7), with a thin colorless wall and a delicate Stieda body. Sporozoites enclose a compact residuum of about 2.5 x 3.7 microns.     

Eimeria saudiensis N. Sp. (Apicomplexa: Eimeriidae) from the Arabian Oryx (Oryx leucoryx) in Saudi Arabia

Oocysts of Eimeria saudiensis n. sp. (Apicomplexa: Eimeriidae) are described from the feces of the Arabian oryx, Oryx leucoryx , from the Riyadh Zoo, Saudi Arabia. The oocysts were ellipsoidal or slightly ovoid, 31.2 times 24.5 (24.3–36.5 times 20.0–27.6) μm with a bilayered wall about 1.7 μm thick. The micropyle was covered by a dome-shaped cap. The oocyst residuum was absent, but tiny polar granules were present. The sporocysts were elongate ovoid, 14.3 times 7.2 (11.5–18.5 times 6.0–9.0) μm, had a Stieda body, but lacked a substiedal body. The sporocyst residuum was present, composed of numerous small granules. The sporozoites were elongate club-shaped, and contained two prominent refractile bodies.