Fine structure of the oocyst walls of Isospora serini and Isospora canaria and excystation of Isospora serini from the canary, Serinus canarius L.

Oocysts of Isospora serini and Isospora canaria, from the canary Serinus canarius, were broken, added to a cell suspension, fixed in Karnovsky's fluid, and studied in the electron microscope. The oocyst wall of each species had an electron-lucent inner layer, a more osmiophilic middle layer and an outer layer of electron-lucent (I. serini) or electron-dense material interspersed with some electron-lucent material (I. canaria). A few, relatively large lipid-like bodies were present in the outer or middle layer of the oocyst wall of I. canaria. As many as 9 membranes were present in the oocyst wall of I. canaria and 3 in that of I. serini. When exposed to a trypsin-sodium taurocholate fluid, sporozoites of I. serini excysted from 5-month-old sporocysts in vitro, but not from sporocysts stored for more than 6 months. No excystation occurred in 15-month-old I. canaria sporocysts. Similarities and differences in excystation between I. serini and other Isospora, Eimeria, and Sarcocystis species are discussed.     

Exogenous stages of Isospora serini (Arag?o)and Isospora canaria sp. n. in the canary (Serinus canarius Linnaeus).

Exogenous stages of Isospora serini (Aragao) and Isospora canaria sp. n. from the canary (Serinus canarius Linnaeus) are described. Oocytes of I. serini are spheroid and average 19.2 times 20.1 mum, while those of I. canaria are larger, more ellipsoid, and average 21.8 times 24.6 mum. No oocyst residuum is present and the oocyst walls of both species are colorless, transparent, and single layered. Sporocysts average 9.4 times 15.2 mum for I. serini and 11.5 times 18.1 mum for I. canaria. The I. canaria sporocyst has a substiedal body, but none was found in I. serini sporocysts. Both species have a spherical sporocyst residuum; this was obscured in the I. serini sporocyst by scattered granules. Living sporozoites of I. canaria average 3.6 times 16.9 mum and have 1 to 3 refractile globules; those of I. serini have 2 globules and average 2.8 times 12.6 mum. A disseminated infection of the mononuclear phagocytes results from administration of I. serini while I. canaria oocysts give rise to a typical coccidian infection restricted to the intestinal epithelium. Asexual stages of I. serini in macrophages are indistinguishable from parasites previously called avian Toxoplasma, Atoxoplasma, and Lankesterella.     

Exogenous Stages of Isospora serini (Aragão) and Isospora canaria sp. n. in the canary (serinus canarius Linnaeus)*

Exogenous stages of Isospora serini (Aragão) and Isospora canaria sp. n. from the canary (Serinus canarius Linnaeus) are described. Oocysts of I. serini are spheroid and average 19.2 × 20.1 μ m, while those of I. canaria are larger, more ellipsoid, and average 21.8 × 24.6 μ m. No oocyst residuum is present and the oocyst walls of both species are colorless, transparent, and single layered. Sporocysts average 9.4 × 15.2 μ m for I. serini and 11.5 × 18.1 μ m for I. canaria. The I. canaria sporocyst has a substiedal body, but none was found in I. serini sporocysts. Both species have a spherical sporocyst residuum; this was obscured in the I. serini sporocyst by scattered granules. Living sporozoites of I. canaria average 3.6 × 16.9 μ m and have 1 to 3 refractile globules; those of I. serini have 2 globules and average 2.8 × 12.6 μ m. A disseminated infection of the mononuclear phagocytes results from administration of I. serini while I. canaria oocysts give rise to a typical coccidian infection restricted to the intestinal epithelium. Asexual stages of I. serini in macrophages are indistinguishable from parasites previously called avian Toxoplasma, Atoxoplasma, and Lankesterella.     

Life cycles of two isopora species in the canary, Serinus canarius Linnaeus

The endogenous stages of Isospora serini Arog?o and Isospora canaria Box are described from experimentally infected canaries, Serinus canarius Linnaeus. Unlike other Coccidia, the first part of the I. serini life cycle takes place in mononuclear phagocytes. Five asexual generations are described from this cell type; 2 additional asexual generations and the sexual stages take place in the intestinal epithelium. Isospora canaria, on the other hand, has a conventional coccidian life cycle in that all of the endogenous stages are in the epithelium of the small intestine, with 3 asexual generations and the sexual generation described in the duodenal epithelium. The 2 species differ in their position relative to the nucleus of the intestinal epithelial cell. Isospora serini is usually on the lumenal side of the nucleus while I. canaria is below the nucleus, toward the basement membrane. The prepatent period is 4-5 days for I. canaria and 9-10 days for I. serini. Patency lasts for 11-13 days in I. canaria infections, but duration of oocyst output is more chronic in I. serini infections, persisting for as long as 231 days. Both species have a diurnal periodicity of oocyst discharge which occurs in late afternoon and evening.     

Life Cycles of Two Isospora Species in the Canary,Serinus canarius Linnaeus*

SYNOPSIS. The endogenous stages of Isospora serini Aragão and Isospora canaria Box are described from experimentally infected canaries, Serinus canarius Linnaeus. Unlike other Coccidia, the first part of the I. serini life cycle takes place in mono-nuclear phagocytes. Five asexual generations are described from this cell type; 2 additional asexual generations and the sexual stages take place in the intestinal epithelium. Isospora canaria, on the other hand, has a conventional coccidian life cycle in that all of the endogenous stages are in the epithelium of the small intestine, with 3 asexual generations and the sexual generation described in the duodenal epithelium. The 2 species differ in their position relative to the nucleus of the intestinal epithelial cell. Isospora serini is usually on the lumenal side of the nucleus while I. canaria is below the nucleus, toward the basement membrane. The prepatent period is 4–5 days for I. canaria and 9–10 days for I. serini. Patency lasts for 11–13 days in I. canaria infections, but duration of oocyst output is more chronic in I. serini infections, persisting for as long as 231 days. Both species have a diurnal periodicity of oocyst discharge which occurs in late afternoon and evening.     

The Structure of the Oocyst and the Excystation Process of Isospora marquardti sp. n. from the Colorado pika, Ochotona princeps

SYNOPSIS. Oocysts of Isospora marquardti sp. n. from the Colorado pika, Ochotona princeps , have spheroid oocysts, 30.5 (23–36) μm in diameter, and ovoid sporocysts, measuring 19.3 × 12.0 (17–22 × 10–14) μm. A polar body, 2 × 4 μm, a spheroid sporocyst residuum (8.3 μm in diameter), a Stieda body, and a distinct substiedal body (3 × 3 μm) are present. A micropyle and an oocyst residuum are absent. Excysted sporozoites, averaging 3.0 × 18.5 (2–4 × 15–20) μm, contain 2 refractile globules, 1 on each side of the nucleus with a prominent nucleolus.
The sporozoite excystation process using a trypsin-sodium taurocholate fluid is described.     

Isospora daphnensis n. sp. (Apicomplexa: Eimeriidae) from the medium ground finch (Geospiza fortis) from the Galapagos Islands

In March and April 1987 fecal samples from 237 nestling birds, including 199 from 85 nest sites of Geospiza fortis, 23 from 12 nest sites of Geospiza scandens, 6 from 2 nest sites of Geospiza magnirostris, and 9 from 2 nest sites of hybrids involving Geospiza fuliginosa and G. fortis, were collected from Daphne Major in the Galapagos archipelago and examined for coccidia. Only 3 of 4 nestlings from 1 nest site of G. fortis (1.5%) had oocysts in their feces. Two of the 3 infected nestlings had concurrent infections of Isospora temeraria, and all 3 nestlings were infected with a new species. Isospora daphnensis n. sp. Sporulated oocysts of I. daphnensis n. sp. are ellipsoidal, 27.3 x 23.6 (22-30 x 20-27) microns; a polar body is present, but no oocyst residuum or micropyle occurs. The oocyst wall, approximately 1.5 microns thick, is composed of a mammillated outer layer and thinner inner layer. Sporocysts are ovoid, 15.2 x 10.2 (15-16 x 9-11) microns and have a nipplelike Stieda body and a small substieda body. The sporocysts contain an irregularly shaped, smoothly contoured residuum with uniform granules and 4 sporozoites with a large refractile body at one end and lying randomly in the sporocysts.     

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.     

Isospora viridanae N. Sp., an Intranuclear Coccidian Parasite from the Canarian Skink, Chalcides viridanus (Lacertilia: Scincidae)

A caryotropic species of coccidium, Isospora viridanae n. sp. (Apicomplexa: Eimeriidae). has been found as a parasite in the Canarian skink, Chalcides viridanus Gravenhorst, 1851, from Tenerife, Spain, and is described here as a new species. Fully sporulaled oocysts of Isospora viridanae are spherical and measure 21.6 (1 7.6–23.4) nm in diameter. Mtcropyle, oocyst residuum and polar granule are ali absent. Oocysts of this coccidian, with a smooth bilayered wall, contain 2 ovoid sporocysts 13.2 (11.7–14.0) by 9.5 8.2–10 5) μm. A sporocyst residuum is present as well as a Stieda body and a substieda body. Most oocysts are found to be at the beginning of sporulanon when excreted and show 2 spherical sporoblasts. Sporulation is completed within 24 to 48 h at 23 × 2°C Sporozoites are 13–14 μm long and are about 2.5–3 μ m wide. Endogenous stages of schizogony and gamogony develop in the nuclei of epithelial cells from the small intestine of the skink. Comparisons with other species of the genus found in lacertilian hosts indicate that it is a new species.     

Excystation of Isospora arctopitheci Rodhain, 1933 with notes on a similiar process in Isospora begemina (Stiles, 1891) Lühe, 1906.

The in vitro excystation process of sporozoites of Isospora arctophitheci Rodhain, 1933 from the titi marmoset Saguinus geoffroyi and of Isospora bigemina (Stiles, 1891) Lühe, 1906 from the bobcat, Lynx rufus is presented. Sporocysts of both species lack a Stieda body and when exposed to a trypsin-sodium taurocholate (pH 7.4) excysting fluid the walls of both collapse in a similar fashion, along apparently predetermined lines. Similarities and differences on excystation between. I. arctopitheci, I. bigemina, and other Isospora, Eimeria, and Sarcocytis species are summarized. Such studies show that 2 distinct patterns of sporozoite excystation have been described to date, and both appear to be related to the structure of the sporocyst.     

Eimeria procyonis sp. n., an Isospora sp., and a Redescription of E. nuttalli Yakimoff and Matikaschwili, 1932 (Protozoa: Eimeriidae) from the American Raccoon (Procyon lotor)

SYNOPSIS. Thirty-two of 48 raccoons examined were infected with a previously undescribed species of Eimeria which is herein named E. procyonis. Of the 32 infected animals, 10 also harbored E. nuttalli and 1 had Isospora sp. oocysts.
The ellipsoid to ovoid oocysts of E. procyonis measured 23.4 × 18.0 (16–29 × 13–24) μm; its sporocysts measured 12.1 × 9.3 (11.5–15 × 7–10) μm, each containing a slightly flattened substiedal body. The sporocyst residuum consisted of numerous scattered granules each ∼1 μm in diameter. The oocyst wall was double-layered. The outer layer appeared rough and pitted, measuring 1.5 μm, except at the micropyle where it was 1 μm thick.
The oocysts of the Isospora sp. measured 16.8 × 13.7 (16–18.5 × 12.5–15.5) μm. The wall consisted of a single layer ∼0.5 μm thick. The sporocysts measured 11.2 × 9.1 (9.5–11.5 × 8–10) μm, and each contained 4 elongate sporozoites. The oocysts of E. nuttalli measured 17.5 × 13.6 (12-21 × 11-15) μm, with a smooth single-layered wall approximately 0.7 μm thick. The sporocysts measured 12.2 × 7.1 (9-13 × 5.5–11) μm. Each sporocyst had a thin, dark, Stieda body and the sporocyst residuum consisted of many fine granules.     

Eimeria cicaki sp. n. and Isospora thavari sp. n. from the house lizard Gehyra mutilata Boulenger in Malaysia.

Oocysts and endogenous stages of new species of Eimeria and Isospora from the house lizard, Gehyra mutilata, are described. The ellipsoid to subspherical 2-layered oocysts of E. cicaki averaged 24.0 X 21.0 mum. Polar granules are present. Micropyle and oocyst residuum are absent. Ellipsoid sporocysts average 12.2 X 9.0 mum. A sporocyst residuum is present, but the Stieda body is absent. Endogenous stages are in epithelial cells of the small intestine. The subspherical single-layered oocysts of I. thavari average 23.8 X 22.8 mum. The polar granule is present; micropyle and oocyst residuum are absent. Ellipsoid sporocysts average 12.8 X 9.4 mum. Stieda body and sporocyst residuum are present. There are endogenous stages in epithelial cells of the small intestine.     

Isospora Normanlevinei N. Sp. and Isospora Coluzzii N. Sp. (Apicomplexa: Eimeriidae) In Emberiza Cirlus (Cirl Bunting) (Passeriformes-Emberizidae)

ABSTRACT. The morphological characteristics of two new species of Isospora observed in Emberiza cirlus (Cirl Bunting) from Italy are reported. the oocysts of Isospora normanlevinei n. sp. are spherical or sub-spherical, with a smooth double-layered wall, and measure 24.2 times 23.7 (21.0-26.5 times 21.5-25.5) μm; each oocyst contains 2 to 10 polar granules. No micropyle or residuum was observed. the piriform sporocysts measure 19.4 times 11.2 (17.0-21.0 times 10.0-12.5) μm and contain a dispersed residuum. the Stieda body is flat; the substiedal body, with scattered clear and dark granules, may be either symmetrical or asymmetrical. the oocysts of I. coluzzii n. sp. are asymmetrical and rounded shape and measure 28.6 times 24.2 (25.0-31.5 times 21.5-26.0) μm. the oocyst has a double-layered wall and 2 to 3 polar granules. Neither micropyle nor residuum is present. the sub-ellipsoidal sporocyst, measuring 18.2 times 10.0(16.5-20.0 times 9.0-11.0) μm, has a dispersed sporocyst residuum. the Stieda complex is symmetrical.     

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.     

Isospora dawadimiensis n. sp. (Protozoa: Eimeriidae) from the Jerboa (Jaculus jaculus) in Saudi Arabia

Isospora dawadimiensis n. sp. is described from the jerboa, Jaculus jaculus, from Dawadimi, Saudi Arabia. Sporulated oocysts of I. dawadimiensis n. sp. were ovoidal or nearly subspherical 22–26.5 times 20.5–22 μm (24.4 times 21.4 μm). Oocyst wall had one layer. Micropyle, oocyst residuum, and polar granule were absent. Sporocysts were ellipsoid 12–16.5 times 9–10.5 μm (14.6 times 9.9 μm). Sporocyst residuum was present. The sporocysts lack a Stieda body. Sporozoites 8–11 times 2–3 μm (10 times 2.6 μm) were sausage-shaped, slightly curved, and tapered at one end.     

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.     

Isospora gallicolumbae sp. n. from Beccari's Ground Dove (Gallicolumba beccarii Salvadori) in Papua New Guinea

SYNOPSIS. Isopora gallicolumbae sp. n. is described from Beccari's ground dove, Gallicolumba beccarii Salvadori, in Papua New Guinea. The ellipsoidal oocysts average 16.0 × 20.0 μm. The oocyst wall is light green, single-layered (∼ 0.8 μm), and becomes distorted within 10 min in sugar solution. Micropyle and oocyst residuum are absent; one polar granule is present. Sporocysts are ovoid, 8.0 × 12.0 μm. with prominent conical Stieda body. Sporocyst residuum is a spherical mass (∼ 5.0 μm) of lighter and darker granules. This is the first species of Isospora reported from a host in the avian order Columbiformes.     

Isospora petrochelidon sp. n. (Protozoa: Eimeriidae) from the Cliff Swallow, Petrochelidon pyrrhonota

SYNOPSIS. Isospora petrochelidon sp. n. (Protozoa: Eimeriidae) is described from the cliff swallow, Petrochelidon pyrrhonota , from Douglas County, Colorado. Oocysts were passed primarily between 4 and 9 p.m. Sporogony was completed by 24 hr in 3% aqueous potassium dichromate at 21–28 C. The ellipsoid to ovoid oocysts averaged 25.2 × 22.2 μm. The oocyst wall was smooth, colorless, and single-layered. The 2 polar granules were disc-shaped, each adjoining a sporocyst. The lemon-shaped sporocysts averaged 18.4 × 10.8 μm. There was no membrane enclosing the sporocyst residuum and sporozoites. Two fledgling Isospora -free swallows fed sporulated oocysts began passing them on day 4 and died with massive infections on day 5. Data from these birds indicate that schizogony occurred during nighttime hours, gametogony during the day. Endogeny was found throughout the intestine distal to the gizzard, with schizogony confined to the convoluted glands of the subvillar mucosa and gametogony to the villar epithelium. Sporocysts released from mechanically ruptured oocysts excysted in 2% pancreatin within 5 min. Sporulated oocysts fed to a pigeon squab, duckling and cockerel produced no infections.     

Ultrastructure of Excystment of Toxoplasma gondii Oocysts*

SYNOPSIS. The excystation of sporozoites from intact Toxoplasma gondii oocysts or mechanically released sporocysts was studied by light and electron microscopy. Both intact oocysts and free sporocysts excysted in 5% bovine bile in 0.9% NaCl solution after 30–60 min incubation at 37 C. Sporozoites were first activated in either intact sporocysts or oocysts within 2–12 min of incubation in bile. Sporozoites escaped from sporocysts through 4 plate-like sutures in the sporocyst wall, and from the oocyst as the oocyst wall ruptured at one or more points.     

Experimental Isospora canis and Isospora felis Infection in Mice,Cats, and Dogs*

SYNOPSIS. Two 6-month-old gnotobiotic dogs and 4 five-day-old dogs became infected but did not shed oocysts within 15 days after ingesting the feline coccidian, Isospora felis. Infection of the dogs was evidenced by the shedding of I. felis oocysts by cats consuming extra-intestinal organs of dogs fed I. felis. Likewise, cats became infected with the canine coccidian, Isospora canis, without producing oocysts. Dogs also became infected after ingesting mice previously fed I. canis oocysts. The prcpatent period for I. canis was slightly shorter in dogs fed infected mice (8–9 days) than in those fed oocysts (9–11 days).