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Neospora caninum (Protozoa: apicomplexa) infections in mice.

Groups of mice were given 0 mg, 4 mg, or 2 mg of methylprednisolone acetate (MPA) 7 days prior to, the day of, and 7 days after subcutaneous inoculation with 0 or 2 x 10(5) tachyzoites of Neospora caninum. Clinical signs of disease were seen only in mice given both MPA and N. caninum tachyzoites. Mice given 4 mg MPA and N. caninum tachyzoites developed severe disseminated neosporosis and most died or were killed when comatose 11-13 days postinoculation (PI). Acute pneumonia, polymyositis, encephalitis, hepatitis, and pancreatitis were the main lesions in these mice. Mice given 2 mg MPA and N. caninum developed mild pneumonia and many mice began showing neurological signs 14 days PI. Neurological signs consisted mainly of pronounced head-tilting and associated impairment of movement. Grossly visible 1-2-mm single or multiple, white areas of discoloration were seen in the brains of many of these mice. Encephalitis, ganglioradiculoneuritis, pneumonia, and polymyositis were the main changes seen in these mice. Tissue cysts of N. caninum were only seen in mice given 2 mg MPA and were first seen 21 days PI. Tissue cysts were 16-34 by 13-29 microns and had a 1.5-3.0-microns-thick cyst wall. Tissue cysts were seen only in the brain. Mice given 4 mg MPA and tachyzoites and host cells that had been frozen for 1 wk did not develop clinical signs of infection, indicating that freezing kills tachyzoites and that viruses or other agents were not involved in the genesis of disease seen in mice given MPA and viable tachyzoites.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Eimeria cryptotis n. sp. (Apicomplexa: Eimeriidae) from the least shrew, Cryptotis parva (Insectivora: Soricidae), in north-central Texas.

From March through November 1987, 14 least shrews, Cryptotis parva (Say), were collected in portions of north-central Texas and examined for coccidian parasites; only 1 (7.1%) was found to be passing oocysts. Eimeria cryptotis n. sp. is described herein as new and represents the only coccidian reported thus far from C. parva. Sporulated oocysts are subspherical, 16.4 x 15.3 (14-18 x 13-17) microns; shape index 1.1 (1.0-1.2) microns. A micropyle and oocyst residuum are absent, but a polar granule is present. The sporocysts are ovoid, 10.6 x 7.0 (9-11 x 6-8) microns; shape index 1.5 (1.4-1.8) microns. Stieda and substieda bodies and a sporocyst residuum are present. The sporozoites are elongate and only 2 could be observed well enough to measure (11.2 x 2.4 and 8.8 x 2.4 microns) because they are normally obscured by the sporocyst residuum. Sporozoites lack refractile bodies and contain a centrally located nucleus. The new species can be distinguished from the majority of insectivore coccidia on the basis of oocyst size.

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Suppression of Nippostrongylus brasiliensis (Nematoda)-induced lysophospholipase activity and peripheral eosinophilia by Eimeria nieschulzi (Apicomplexa).

Interspecific interactions between Nippostrongylus brasiliensis and Eimeria nieschulzi were studied by measuring fecal lysophospholipase (LYPH) activity and relative numbers of peripheral eosinophils in rats singly or concurrently infected with one or both parasite species. Three groups of 10 rats each were inoculated with 2 X 10(3) N. brasiliensis L3 larvae and/or 5 X 10(5) E. nieschulzi sporulated oocysts. Groups 1 and 2 were infected with E. nieschulzi or N. brasiliensis, respectively. Group 3 rats were infected first with N. brasiliensis, followed on day 8 postinoculation (PI) with E. nieschulzi. Each rat served as its own control. Results revealed LYPH levels rose steadily in Group 2 rats, reaching significant peaks on days 10 and 12 PI before decreasing to control levels. Lysophospholipase activity in Groups 1 and 3, however, did not differ from control values. Group 2 rats also demonstrated peripheral eosinophilia, with peak values occurring on days 10, 12, 14, and 16 PI, while rats in Groups 1 and 3 exhibited no eosinophilia. These results demonstrate that E. nieschulzi suppressed intestinal LYPH activity and relative peripheral eosinophilia and demonstrate that a host's immune response to a single parasite may be significantly altered when a second parasite species is present.

Animals↗

B-lymphocyte responses in the large intestine and mesenteric lymph nodes of mice infected with Eimeria falciformis (Apicomplexa).

B-cell responses of 3 immunoglobulin isotypes (IgA, IgG, and IgM) were investigated in the large intestine and mesenteric lymph nodes (MLN) of naive or immune mice after inoculation of oocysts of Eimeria falciformis. Primary and anamnestic IgA and IgG lymphocyte responses to E. falciformis occurred in the large intestine of nonimmune and immune mice, respectively. IgA-containing lymphocytes (IgAc) were the largest population of responding B cells in the large intestine. In infected mice, IgAc accumulated in the apical portion of the lamina propria, whereas IgG-containing lymphocytes (IgGc) were more numerous at the base of the lamina propria. No significant increase in the number of IgM-containing lymphocytes (IgMc) was observed in the lamina propria of the large intestine. Primary but no anamnestic B-cell responses occurred in the MLN, and immune mice actually had reduced numbers of IgAc and IgGc in the MLN when compared with naive mice. IgGc were the largest population of responding B cells in the MLN. Thus, IgAc appear to accumulate preferentially at the site of parasite development, whereas IgGc are primarily localized deeper in the lamina propria of the large intestine and in the draining lymph nodes of mice infected with E. falciformis.

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Fatal Caryospora bigenetica (Apicomplexa: Eimeriidae) infections in cotton rats, Sigmodon hispidus.

Four groups of cotton rats, Sigmodon hispidus, were shown to be suitable secondary hosts for the viperid coccidium, Caryospora bigenetica, following oral inoculation of a mixture of oocysts and sporocysts. Swelling of the face, ears, and scrota and hemorrhagic ears were the predominant clinical signs and some cotton rats died in 3 of 4 experiments. Developmental stages of C. bigenetica were found in connective tissue components of the ear, nose, cheeks, anal skin, scrotum, and penile sheath of all cotton rats in which these tissues were examined. Additionally, developmental stages of C. bigenetica were found in connective tissue components of the following tissues examined from some cotton rats: tongue, lung, testicle, epididymis, rectum, base of the tail, footpad, and bone marrow. The present study shows that C. bigenetica can be pathogenic for cotton rats and demonstrates many new anatomic sites for developmental stages of this parasite in the secondary host.

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The taxonomy of Sarcocystis (Protozoa, Apicomplexa) species.

The taxonomy of the heteroxenous apicomplexan protozoan genus Sarcocystis was reviewed, and a list of 122 species with their synonyms and hosts given. Both definitive and intermediate hosts are known for only 56 species. The fine structure of the sarcocyst wall may change with age and is not considered necessarily satisfactory for separating species. Specificity for the intermediate host is not narrow for all species. Earlier work on transmission of the parasite from one intermediate host to another should be repeated in the light of present knowledge of the life cycle of species of Sarcocystis.

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[Sarcocystis alceslatrans (Apicomplexa) from a palaearctic elk (Ruminantia)].

Sarcocysts from a palaearctic moose have been studied for the first time by light and electron microscopy. Because of coincidences in morphology of the cysts, especially of the cyst wall structure and the cystozoides, with the North American species Sarcocystis alceslatrans DUBEY, 1980, they are assigned to this species.

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In vitro excystation of Sarcocystis capracanis, Sarcocystis cruzi and Sarcocystis tenella (Apicomplexa).

Improved rates of in vitro excystation of sporozoites from sporocysts of Sarcocystis capracanis, Sarcocystis cruzi, and Sarcocystis tenella were obtained by pretreating sporocysts with an aqueous sodium hypochlorite (NaOCl) solution followed by incubation in excysting fluid (EF). After pretreatment with NaOCl, sporocysts were washed 4 times in Hanks' balanced salt solution and then incubated in various EF (pH 7.4) at 38.5 C in 5% CO2-95% air. Maximum rates of excystation (free sporozoites/(sporozoites in sporocysts + free sporozoites) X 100) for all 3 species of Sarcocystis occurred at 4 hr after incubation in EF. These rates were 17% for S. capracanis after incubation in EF containing 2% trypsin + 10% caprine bile; 90% for S. cruzi in 2% trypsin + 10% bovine bile; and 20% for S. tenella in 2% trypsin + 10% caprine bile. Only a 40% excystation rate occurred in sporocysts of S. cruzi that had been stored previously for 14 days in aqueous potassium dichromate. Excysted sporozoites of S. capracanis, S. cruzi, and S. tenella penetrated and developed to mature meronts in bovine pulmonary artery endothelial cells or bovine monocytes.

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[Current conception of the Sarcosporidia (Sarcocystis, Eimeriidae, Sporozoa, Apicomplexa): their morphofunctional organization, life cycle and practical importance].

Advances in sarcosporidian research within the latest decade are critically reviewed and analysed in respect to some recent cytological findings of the author and her colleagues on the subject. Three rather than two morpho-functional cell types (metrocytes and merozoites) are distinguished within the sarcocyst, the third one being the intermediate cell. Division by endodyogeny in the cyst of Sarcocystis is very likely confined to the latter cell type. The pattern of nuclear chromatin and the constancy in DNA value per nucleus in the cystic merozoites, revealed by flow cytometry, is rather indicative of their incapability of dividing within the cyst, i.e. in the intermediate host. This enabled us to consider these merozoites as homologs of coccidian gamonts. The obvious differences between cysts and cystic stages in Sarcocystis and Toxoplasma may account in part for the rare, if any, reported cases of congenital sarcocystosis in the intermediate host.

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Coccidia (Apicomplexa: Eimeriidae) from the subterranean rodent Ctenomys opimus Wagner (Ctenomyidae) from Bolivia, South America.

Of 35 tuco-tucos (Ctenomys opimus) collected in Bolivia, South America, 31 (88%) had eimerian oocysts in their feces at the time they were examined. Eighteen (58%) of the 31 infected animals were concurrently infected with 2 or 3 eimerian species. Four species of Eimeria were recovered and are described as new species based on the characteristics of sporulated oocysts. Oocysts of Eimeria granifera n. sp. were ellipsoidal, 21.1 x 17.2 (15-26 x 11-20) micron with sporocysts ovoidal, 11.3 x 7.1 (8-14 x 5-9) micron. Oocysts of Eimeria montuosi n. sp. were spheroidal, 24.2 x 22.0 (21-28 x 18-25) micron with sporocysts ovoidal, 10.5 x 7.3 (8-14 x 6-9) micron. Oocysts of Eimeria opimi n. sp. were spheroidal to subspheroidal, 24.3 x 21.8 (18-29 x 15-26) micron with sporocysts ovoidal, 11.6 x 7.6 (10-13 x 6-9) micron. Oocysts of Eimeria oruroensis n. sp. were spheroidal to subspheroidal, 27.3 x 23.6 (23-32 x 20-28) micron with sporocysts ovoidal, 13.2 x 8.6 (10-16 x 8-11) micron.

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Eimeria maxima (Apicomplexa): a comparison of sporozoite transport in naive and immune chickens.

This study compared the early stages of infection in naive and immune chickens infected with Eimeria maxima. An immunoperoxidase stain was developed and used to detect sporozoites and early schizonts in tissue sections of intestinal epithelium. A significantly higher proportion of sporozoites was present in the crypts of naive chickens, 48 hr postinoculation of oocysts, compared to immune chickens. Sporozoites in immune birds tended to remain in the lamina propria rather than migrate to the crypts. Sporozoites were found within intraepithelial lymphocytes (IEL's) in the epithelium, the lamina propria, and the crypts of both naive and immune chickens. Parasites in IEL's of immune birds at the ultrastructural level and there were no apparent morphological abnormalities. Livers and spleens, of both immune and naive chickens that had been inoculated with Eimeria maxima, produced patent infections when fed to susceptible chickens. Infections could be transferred up to 72 hr post-inoculation of the donor birds. Peak oocyst production in the recipient birds occurred 7-8 days after the transfers. This time period approximates the prepatent period in a natural infection and thus implies that the extraintestinal stage was a sporozoite.

Analysis of Variance↗

Coccidian parasites (Apicomplexa: Eimeriidae) from insectivores. III. Seven new species in shrews (Soricidae: Soricinae) from Canada, Japan, and the United States.

Since May 1979, 458 shrews (Blarina sp. and Sorex spp.) representing 20 species collected in Canada, Japan, and the United States were examined for coccidia; 110 (24%) had oocysts in their feces, including 8 of 21 (38%) B. brevicauda from Massachusetts, Ohio, Pennsylvania, and Vermont; 2 of 7 (29%) S. caecutiens from Hokkaido and Honshu; 14 of 63 (22%) S. cinereus from Colorado, New Mexico, Pennsylvania, Vermont, Manitoba, and Ontario; 3 of 7 (43%) S. fontinalis from Pennsylvania; 11 of 16 (69%) S. fumeus from Massachusetts, Minnesota, Pennsylvania, Vermont, and Ontario; 1 of 4 (25%) S. haydeni from Minnesota; 6 of 8 (75%) S. longirostris from Florida and Virginia; 1 of 2 (50%) S. ornatus from California; 5 of 12 (42%) S. pacificus from California and Oregon; 13 of 41 (32%) S. palustris from California, Colorado, and New Mexico; 1 of 2 (50%) S. tenellus from California; 11 of 105 (10%) S. trowbridgii from California, Oregon, and Washington; 10 of 48 (21%) S. unguiculatus from Hokkaido; and 24 of 112 (21%) S. vagrans from Arizona, California, Colorado, New Mexico, Oregon, and Washington. The following coccidians were identified from infected shrews: Eimeria brevicauda n. sp. from B. brevicauda; Eimeria fumeus n. sp. from S. fumeus, S. pacificus, S. unguiculatus, and S. vagrans; Eimeria inyoni n. sp. from S. tenellus; Eimeria palustris n. sp. from S. cinereus, S. fontinalis, S. fumeus, S. haydeni, S. longirostris, S. ornatus, S. pacificus, S. palustris, S. tenellus, S. trowbridgii, and S. vagrans; Eimeria vagrantis n. sp. from S. fumeus, S. trowbridgii, and S. vagrans; Isospora brevicauda n. sp. from B. brevicauda; and Isospora palustris n. sp. from S. pacificus, S. palustris, S. trowbridgii, S. unguiculatus, and S. vagrans. The world literature on coccidian parasites of shrews (16 eimerians and 3 isosporans exclusive of the 7 new species described here) is reviewed.

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Suppression of peripheral eosinophilia by the coccidium Eimeria nieschulzi (Apicomplexa: Eimeriidae) in experimentally infected rats.

Four groups of 60 rats each were used to examine interspecific interactions between Eimeria nieschulzi and Nippostrongylus brasiliensis. Rats in group 1 served as uninoculated controls. Group 2 rats were each injected subcutaneously with 2.0 X 10(3) L3 larvae of N. brasiliensis. Group 3 rats were each inoculated per os with 2.5 X 10(5) sporulated oocysts of E. nieschulzi. Rats in group 4 were first infected with 2.0 X 10(3) larvae of N. brasiliensis and, at 8 days postinoculation, with 2.5 X 10(5) oocysts of E. nieschulzi. Ten animals from groups 1-3 were sacrificed at 4-day intervals postinoculation and group 4 rats were sacrificed at 4-day intervals beginning after the secondary infection. Blood smears were prepared from each animal to determine differential blood cell counts, bone marrow was examined at the times of peak infection for absolute and relative numbers of eosinophils, portions of the duodenum and jejunum were examined histologically for mast cells, and feces obtained from the cecum and large intestine were examined for ova/gram of feces. Results revealed that relative numbers of peripheral neutrophils and monocytes became elevated during the course of infection for all infected animals, and rats infected with the helminth only also had elevated eosinophil levels. However, rats infected singly with E. nieschulzi, or concurrently with the coccidium and helminth, had peripheral eosinophil levels that were not significantly different from controls.(ABSTRACT TRUNCATED AT 250 WORDS)

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Immunity to Eimeria separata (Apicomplexa: Eimeriina): expose-and-challenge studies in rats.

Oocyst production of Eimeria separata of the rat was determined in initial and challenge infections. The total number of oocysts produced was no higher at 100 oocysts than at 1,000, 10,000, or 100,000 in initial infections. In 2 experiments, the reproductive index with 100 oocysts was 17,000 and 34,000, respectively. On challenge with 10,000 oocysts, there was 91% protection at 100 oocysts. There was slightly lower protection at 10,000 and 100,000 oocysts, 72% and 81%, respectively, and t-values suggest lower protection at these higher inocula.

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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.

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Proteins and antigens of merozoites and sporozoites of Eimeria bovis (Apicomplexa).

Proteins and antigens of first-generation merozoites and sporozoites of Eimeria bovis were examined using standard sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), Western blotting and lactoperoxidase iodination procedures. SDS-PAGE gels revealed both common and unique protein bands in merozoite and sporozoite extracts, ranging in molecular weight (Mr) from 15,000 to 215,000. Nitrocellulose immunoblots of separated proteins, when probed with sera obtained from immunized calves, revealed numerous IgG-binding antigens of Mr 18,000 to 180,000 in merozoites and Mr 28,000 to approximately 118,000 in sporozoites. Although merozoite and sporozoite preparations each contained antigens of different molecular weights, 4 antigens had the same migratory distance in both preparations (Mr 58,000, 70,000, 83,000, 98,000). Of 3 types of immune sera used to probe immunoblots, serum taken from a calf that had been inoculated with oocysts of E. bovis and boosted 10 wk later by subcutaneous injection with 2 X 10(7) live merozoites emulsified in Freund's complete adjuvant consistently identified and reacted more intensely with more antigens of merozoites and sporozoites than the other immune sera tested. Autoradiographic analysis of radioiodinated parasites revealed major surface proteins on merozoites of between 15,000 and 18,000 Mr and 3 surface proteins on sporozoites of Mr 28,000, 77,000, and 183,000. All but the 183,000 protein elicited an IgG antibody response in the host.

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