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Ultrastructure of developing Isospora suis in cultured cells.

The ultrastructure of Isospora suis sporozoites, type-1 meronts, and type-1 merozoites was examined, using transmission electron microscopy of infected cultured cells. The ultrastructure of sporozoites and type-1 merozoites was similar. Each possessed trimembranous pellicles, subpellicular microtubules, a conoid, anterior and posterior polar rings, rhoptries, micronemes, a single vesicular nucleus, tubular mitochondria, Golgi complexes, ribosomes endoplasmic reticula, inactive micropores, amylopectin bodies, lipid bodies, dense bodies, and crystalloid bodies. Merozoites were produced by endodyogeny. Ultrastructural events associated with merozoite production by type-1 meronts are described.

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Double alteration of the small intestine in conventional and gnotobiotic piglets experimentally infected with the coccidium Isospora suis (Apicomplexa, Eimeriidae).

The pathogenicity of the coccidium Isospora suis was studied in 42 conventional and 26 gnotobiotic piglets at 1-13 days post infection (DPI). The prepatent period of the coccidium I. suis was 4.5-5 days. Only in one experiment in conventional piglets the prepatent period of 3.5 days was recorded. At least 9 days pathogenesis of I. suis was recorded during which double alteration of the small intestine occurred. The first alteration was found at 3-4 DPI. The second phase of alterative changes was observed at 8-9 DPI. In comparison with gnotobiotic piglets in conventional piglets clinical signs and pathological changes used to appear one day sooner but reparation and regeneration of alterative changes was quicker. In the first phase of the infection (3-7 or 8 DPI) the heaviest pathological changes were in the posterior jejunum. In the second phase of the infection (8 or 9-12 DPI) the heaviest changes were in the middle jejunum. In short period repeated lesions of mucosal barrier of the small intestine multiply the pathogenicity of I. suis and rank I. suis in front place among other enteropathogens of nursing piglets.

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[Ultrastructure of chronic reticuloendothelial forms of Isospora of sparrows].

Mononucleated reticulo-endothelial stages of Isospora from Sparrows were studied by T. E. M. during the winter period when the extra intestinal infection is chronic. Results show a very slow evolution from merozoite to trophozoite stage, a transformation of the parasite nucleus and a characteristic reaction of the host cell. These observations are discussed in relation to what is known of hypnozoite and chronic schizogony of Plasmodium.

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Isospora manchacensis n. sp., an intranuclear coccidian from the Louisiana ground skink, Scincella lateralis (Say, 1823) (Lacertilia: Scincidae).

Isospora manchacensis n. sp. is described from ground skinks, Scincella lateralis (Say, 1823) from Louisiana. Overall prevalence at 6 sites near Lake Ponchartrain was 43.1% (59/137) and ranged from 8% (1/13) to as high as 60% (6/10). Endogenous stages develop inside the nuclei of epithelial cells in the small intestine. Infected hypertrophic nuclei migrate from the basal lamina of the host cell to the luminal striated border. Oocysts in freshly passed fecal pellets usually contain a single contracted sporont that divides to form 2 sporoblasts. These undergo a brief pyramid stage followed by sporulation within 45-50 hr. Sporulated oocysts have a single-layered wall and measure 25.0 X 22.6 (20.0-28.9 X 18.6-26.0) micron. The lemon-shaped sporocysts measure 12.8 X 10.2 (11.1-15.2 X 9.0-11.0) micron and contain a Steida body, a spherical to oval substeida body, and a dispersed, granular sporocyst residuum. Prepatent periods in skinks fed 700 and 1,400 oocysts ranged from 24 to 32 days. Experimentally infected skinks produced large numbers of oocysts continuously during the 3-4 wk they were monitored after the onset of patency, but exhibited no signs of disease. Experimental doses of 200 oocysts failed to produce infections in skinks monitored for as long as 7 wk.

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Effects of experimentally induced Isospora suis infection on morbidity, mortality, and weight gains in nursing pigs.

Forty-nine pigs from 6 litters were inoculated at 3 days of age with 300,000 sporulated oocysts of Isospora suis to determine the effects of neonatal coccidiosis on morbidity, mortality, and weight gain in nursing pigs. Fifty-one control pigs from 6 litters were not inoculated. Three to 5 days after inoculation, the inoculated pigs developed a nonhemorrhagic diarrhea, generally lasting 6 to 10 days, that lead to visible dehydration, loss of condition, and 20.4% mortality. Control pigs did not die or develop coccidiosis during the 3-week study and had significantly (P less than 0.05) greater body weights at 7, 14, and 21 days of age than did the inoculated pigs.

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Control of Isospora suis-induced coccidiosis on a swine farm.

Results of a program designed to control neonatal porcine coccidiosis on a total confinement, farrow-to-finish swine farm are reported. The control program consisted of washing, phenol disinfection, and steam cleaning of farrowing houses and treatment of sows with amprolium HCl before and after farrowing. Before initiation of the control program, 88.9% of the sows examined in the farrowing house were negative for coccidian oocysts, 9.9% were positive for Eimeria spp, and 1.2% were positive for Isospora suis. Most pigs nursing on sows before initiation of the control program had diarrhea at 5 to 10 days of age, which led to dehydration and weight loss. Morbidity was high, and mortality was moderate. Composite fecal samples from these litters were all positive (100%) for I suis. After initiation of the control program, 99.6% of the sows examined in the farrowing house were negative for coccidian oocysts and 0.4% were positive for Eimeria spp. Clinical signs of coccidiosis were rarely present in nursing pigs examined after the control program was initiated; however, I suis was still present in 19.8% of the composite fecal samples from pigs examined. An association between oocyst production in sows and I suis infections in pigs was not found in the present study. Oocysts of Eimeria spp were not found in the feces from the pigs.

Amprolium↗

[Cryptosporidium sp. and Isospora suis in swine in Italy].

The presence of Cryptosporidium sp. and Isospora suis is reported for the first time in piglets in Italy, in the province of Mantova: the first one has been found in 1 and the second one in 39 out of 200 litters. Two piglets, respectively 20-day-old and 24-day-old, were experimentally infected with Cryptosporidium sp. oocysts from naturally infected calves, without clinical signs.

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Canine coccidiosis attributed to an Isospora ohioensis-like organism: a case report.

Coccidiosis due to an Isospora ohioensis-like organism was diagnosed in a 10-week-old pup. The pup had diarrhea, dehydration, and weight loss before it died 7 days after onset of clinical signs. Lesions were limited to the intestinal tract. Beginning in the distal end of the small intestine and extending through cecum and colon were mild histiocytic proliferation in the lamina propria and multifocal cryptitis. Numerous meronts and gamonts of an unidentified coccidium were in or near the lesions. Parasites were in the villous epithelium, lamina propria, and intestinal glands of the distal one-half of the small intestine, cecum, and colon. It was concluded that infection was due to an unidentified coccidium with oocysts structurally similar to those of I ohioensis.

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Coccidiosis in swine: dose and age response to Isospora suis.

Coccidiosis is a disease of the young piglet due to infection with Isospora suis and is characterized by diarrhea which is nonresponsive to antibacterial therapy. There is variable morbidity and mortality. Piglets develop a more severe clinical illness and enteritis when infected with I. suis at one to three days of age than when infected at two weeks of age. Microscopic lesions range from villous atrophy and mild erosion to severe fibrinonecrotic enteritis.

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Sporogony of Isospora suis Biester, 1934 of swine.

Sporogony of Isospora suis of swine is described. Unsporulated oocysts were collected immediately after defecation, mixed in 2.5% potassium dichromate solution, incubated at various temperatures, examined at different time intervals, and classified as to stage of development. The uninucleate sporont underwent nuclear division and became binucleated, and then divided to form two round, uninucleate sporoblasts. Binucleate sporoblasts elongated to form binucleate sporocysts, each of which then gave rise to four sporozoites. Sporulation was completed within 56 hr at 20 C, 40 hr at 25 C, 16 hr at 30 C, and 12 hr at 37 C. No sporulation occurred at 4, 40, 42, 45, or 50 C. The thermal death point was between 37 and 40 C.

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Demonstration of immunity against Isospora suis in swine.

Piglets naturally exposed or experimentally infected with Isospora suis oocysts were given challenge doses of oocysts to determine the extent of development of immune resistance. Piglets in both studies shed low numbers of, or no detectable oocysts, following challenge. Administration of methylprednisolone acetate failed to induce oocyst shedding in previously infected piglets. Piglets rechallenged with I. suis following steroid injections also failed to shed significant numbers of oocysts suggesting development of immunity to reinfection.

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Endogenous development of the swine coccidium, Isospora suis Biester 1934.

The endogenous development of Isospora suis Biester 1934 is described in piglets inoculated with 150,000 or 200,000 sporulated oocysts. Endogenous stages developed within villous epithelial cells throughout the small intestine. Two distinct types of meronts were seen in tissue sections. Type I meronts, which were seen at 3 days postinoculation, were binucleate, elongate, and 10.5 by 4.7 micron. They produced two to 14 Type I merozoites per parasitophorous vacuole. Type I merozoites were 10.0 by 3.6 micron. They produced two to 14 Type I merozoites per parasitophorous vacuole. Type I merozoites were 10.0 by 3.6 micron. Type II meronts, which were seen at 4 days postinoculation, were elongate and contained three to 12 nuclei. Type II meronts were 11.4 by 5.3 micron, and one to four were found per parasitophorous vacuole. Type II merozoites were 6.3 by 2.1 micron, and three to 16 were found per parasitophorous vacuole. The peak of asexual development occurred 4 days postinoculation. Fully developed microgamonts, macrogamonts, and oocysts were seen 5 days postinoculation. The prepatent period was 5 days, and the patent period was 5 to 8 days. No extraintestinal stages were seen.

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A description of Isospora amphiboluri (Apicomplexa: Eimeriidae) from the inland bearded dragon, Pogona vitticeps (Sauria: Agamidae).

Fecal samples from 50 captive inland bearded dragons, Pogona vitticeps (Ahl, 1926), bred in California, were examined for coccidian parasites. Sixteen (32%) of the lizards were found to be passing oocysts of Isospora amphiboluri Cannon, 1967, previously described from bearded dragons Pogona barbata (Cuvier, 1829) from Australia. Sporulated oocytes were spherical to subspherical, 25.3 x 25.1 (23-26 x 23-26) microns, with a shape index (length/width) of 1.0 (1.0-1.1). A micropyle, oocyst residuum, and polar granule were absent. Sporocyts were ovoidal, 17.0 x 11.4 (16-18 x 11-12) microns, with a shape index of 1.5 (1.4-1.7). A sporocyst residuum, Stieda, and substieda bodies were present, but parastieda bodies were absent. Sporozoites were elongated, 13.9 x 3.5 (12-15 x 3-4) microns in situ, containing spherical anterior and posterior refractile bodies. The occurrence of I. amphiboluri in P. vitticeps is a new host and geographic record for the parasite. Photomicrographs of the oocysts and endogenous life cycle stages of I. amphiboluri are presented for the first time.

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Shedding of oocysts in piglets experimentally infected with Isospora suis.

Forty-seven piglets were inoculated with doses of 100 to 50,000 sporulated oocysts of Isospora suis. After 5-7 days oocysts were found in faeces. The patent period extended from 8 to 16 days. The shedding of oocysts showed a cyclic pattern with 2-3 peaks separated by intervals of approximately 5 days. Subpatent periods were often seen between the peaks. The level of oocyst shedding during the initial days of the patent period reflected, to some extent, the inoculation dose. However, a maximum of OPG at the 100,000 level was observed among one or more piglets from all groups, regardless of the inoculation dose. Among the majority of piglets inoculated with more than 100 oocysts, the highest OPG-figures were observed in the first peak of the cyclic pattern. Unlike this, the maximum of OPG was observed in the second peak of the cycle among 6 of the 7 piglets inoculated with 100 oocytes only. The triphasic pattern was most pronounced in the low dose group. The marked upscaling of oocyst production, as particularly registered in the low dosed groups, seams to explain at least part of the problems met under practical conditions, when trying to eliminate the transmission of oocytes between successive litters in the farrowing boxes. The cyclic excretion pattern and an apparent absence of autoinfection may indicate that the development of I. suis in the host includes several oocyte producing generations descending from the same initial infection. The presence of subpatent periods can probably explain the marked variation in OPG, as they are often recorded when examining faecal samples from piglets, even when the samples are originating from the same litter.

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Isospora ohioensis sp. n. proposed for I. rivolta of the dog.

A new name, Isospora ohioensis, is proposed for I. rivolta-like oocysts from dog feces. The name I. rivolta is retained for the cat species. The structure of the oocysts of I. ohioensis is described from a cloned culture of oocysts. Oocysts are 24 by 21 mum, sporocysts 17 by 12 mum, and sporozoites 10 by 3 mum. In cross-infection experiments none of 6 coccidia-free dogs shed oocysts after consuming feline I. rivolota, and none of 4 coccidia-free cat shed oocysts after eating canine I. rivolta.

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[Light and electron microscope studies on cysts of sarcocystis fusiformis in the muscles of calves infected experimentally with oocysts and sporocysts of isospora hominis Railliet et Lucet, 1891. 1. The development of cyst and cyst wall (author's transl)].

In several experiments young calves were infected with isospora hominis sporocysts excreted by human males who had ingested raw beef containing Sarcocystis fusiformis cysts. On the 29th, 48th, 62nd and 98 day p.i. the calves were killed and the development of S. fusiformis cysts in muscle cells was studied by light and electron microscopy. Beginning with the 48th day p.i. numerous cysts containing small numbers of metrocytes only were observed. The cysts developed from a parasitophorous vacuole within the host cells. At first this parasitophorous vacuole was limited by a single unit membrane, which soon became thickened at numberous places inside of the vacuole by osmiophilic material. This complex is called primary wall (equal to Primärhülle), reaching a thickness of up to 32 nm in old cysts. This primary wall became regularly folded. forming palisade-like protrusions. On the 62nd day after inoculation these protrusions measured about 2-3 mum in length and 0.5-0.7 mum in diameter. In light microscopy the combined protrusions had the appearance of a radially striated "thick wall", because of their close proximity to each other.

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[Light and electron microscope studies on cysts of Sarcocystis fusiformis in the muscles of calves infected experimentally with oocysts and sporocysts of Isospora hominis Railliet et Lucet, 1891. 2. Fine structure of metrocytes and merozoites (author's transl)].

In several experiments calves were infected with sporocysts of Isospora hominis thus producing "thick-walled" cysts of Sarcocystis fusiformis in muscles. Following the growth of the cyst the development of the cyst stages (metrocytes, merozoites) was studied by electron microscopy. Cyst formation began about 40 days p.i. from a parasitophorous vacuole containing exclusively ovoid metrocytes. On the 62nd day p.i. mainly metrocytes and a few banana-shaped merozoites were present within the cysts, whereas on the 98th day p.i. only merozoites, i.e. infectious stages, were observed. The metrocytes were surrounded directly by the amorphous ground substance of the cyst's interior, but the merozoites were arranged relatively loose within chamber-like hollows of the ground substance. The metrocytes are globular cells, about 6-7 mum by 4. 5 mum. The typical three-layered pellicle had a few invaginations and several micropores, which seem to ingest numerous small vesicles from the interior of the cyst. These cells posses a conoid, polar ring with 22 anchored supellicular microtubules, several rhoptries and micronemes, a glogi complex anterior to the large nucleus. The nucleus has a spherical nucleolus consisting of granular and fibrillar zones. Chromosomal structres were seen in two different stages: large dense plaques (condensed stage), and as small dense granules of 300-400 a diameter, arranged sperically within the karyoplasm ( extended stage). The nuclear pores show the typical eight-fold symmetry known from other protozoa and numerous metazoa. The rought endoplasmic reticulum is very prominent within these cells as well as the tubular mitochondria...

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[Light and electron microscope studies on cysts of Sarcocystis fusiformis in the muscles of calves infected experimentally with oocysts and sporocysts of the large form of Isospora bigemina from dogs. 1. The development of cysts and "cyst wall" (author's transl)].

In several experiments young calves were infected with Isospora bigemina (large form) sporocysts excreted by dogs which had been fed with raw beef containing Sarcocystis fusiformis cysts. On the 27th, 34th, 62nd, 76th and 150th day p.i. the calves were killed and the development of S. fusiformis cysts in muscles cells was studied by light and electron microscopy. On the 27th day p.i. in light microscope preparations numerous schizonts, merozoites and endodyogeny-stages were seen in various organs, such as the liver, lung, kidney, heart, small intestine, esophagus, skeletal muscles, diaphragm, cerebrum, and cerebellum. The merozoites measured 7-8 mug by 2-3 mum. Beginning with the 34th day p.i. numerous cysts containing small numbers of metrocytes only were observed in electron microscopy, too. The cysts developed from a parasitophorous vacuole within the host cells. At first this vacuole was limited by a single unit membrane, which soon became thickened byosmiophilic material at numerous places inside of the vacuole. This complex, called primary wall (= Primärhülle), reached a thickness of up to 200-250 A in all cysts. During growth of the cyst this primary wall became regularly folded forming alternating long and short club-shaped protrusions. The longer protrusions were about 0.6 mum long and 0.2-0.3 mum in diameter, whereas the short protrusions were of about 0.13 mum in length. In light microscopy the combined protrusions had the appearance of a very thin cyst wall because of their small size and their close proximity to each other. Later, all protrusions became longer with a maximum of about 3 mum in length without any change in the diameter. Yet, from the 76th day p.i. these protrusions appeared no longer straight, but they became folded over, following a course along the surface of the cyst. Evidently the protrusions did not increase in number after their initial formation, for the distance between them became greater in older cysts. No fibrillar elements were seen within these protrusions which probably accounts for the folding over. The zone of the superficial folded protrusions was not thicker than 1 mum so that in light microscope even the old cysts appeared as relatively thin walled. The interior of the original electron-pale parasitophorous vacuole bacame progressively condensed during the growth of the cyst. There was development of an amorphous ground substance, containing fine fibrils and granules. The ground substance became divided into thin speta (not visible with the light microscope) forming numerous changer-like hollows. The parasites were very closely packed within these hollows. At the beginning of the cyst formation only metrocytes were found within the young cysts, whereas on the 76th day p.i. and later only the infectious merozoites were present. It is therefore concluded that about 3 months after inoculation of calves with sporocysts of the large form I. bigemina from dogs the cysts are fully differentiated, thus being ready for a new transmission...

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