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Development of Caryospora simplex (Apicomplexa: Eimeriidae) from sporozoites to oocysts in human embryonic lung cell culture.

Leighton tubes containing monolayers of human embryonic lung cells were inoculated with 70,000 or 30,000 sporozoites of the viperid coccidium Caryospora simplex and examined at 1, 2, 4, 6, 8, 10, 12, 14, 16, and 18 days post-inoculation (PI). By day 1 PI, sporozoites had penetrated cells and were within parasitophorous vacuoles. Most sporozoites became spherical and then underwent karyokinesis several times between days 2 and 6 PI. Mature Type I meronts were found on days 6-16 PI and contained 8 to 22 short, stout merozoites. Mature Type II meronts were present on days 10-18 PI and contained 8 to 22 long, slender merozoites. Developing gamonts (undifferentiated sexual stages) were observed on days 14 and 16 PI. Mature micro- and macrogametes and thin-walled unsporulated oocysts were present on days 16 and 18 PI. Attempts to sporulate oocysts in tissue culture medium or in a 2.5% (w/v) aqueous solution of K2Cr2O7 at 25 degrees C and 37 degrees C were unsuccessful; only a few oocysts developed to the contracted sporont stage. Four Swiss-Webster mice injected intraperitoneally with merozoites obtained from Leighton tubes on day 10 PI did not acquire infections. This is the second coccidium reported to complete its entire development, from sporozoite to oocyst, in cell culture.

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Taxonomy and review of the coccidian genus Cryptosporidium (protozoa, apicomplexa).

Reports of Cryptosporidium in various hosts and cross-transmission experiments are reviewed. Cryptosporidium has been found in mammals (Primates, Artiodactyla, Perissodactyla , Carnivora, Lagomorpha, and Rodentia), birds, reptiles, and fish. The only cross-transmission attempts that have been made have been from mammals to other mammals and to a few birds. Names have been given to 19 "species," but it is concluded that only four of these should be considered valid at present. These are: C. muris Tyzzer, 1907 in mammals, C. meleagridis Slavin , 1955 in birds, C. crotali Triffit , 1925 in reptiles, and C. nasorum Hoover , Hoerr , Carlton , Hinsman & Ferguson, 1981 in fish.

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Coccidian parasites (Apicomplexa: Eimeriidae) from insectivores: new species from shrew moles (Talpidae) in the United States.

All of 18 shrew moles, Neurotrichus gibbsii, collected in Oregon and Washington were infected with one or more species of coccidia. Three eimerians and one isosporan were identified and described as new species. Sporulated oocysts of Eimeria heterocapita n. sp. were subspheroid to ellipsoid, 25.5 X 21.4 (23-27 X 18-23) micron. A membranous, cap-like structure was present at one pole of the oocyst, but a micropyle, oocyst residuum, and polar body were absent. Ovoid sporocysts were 13.6 X 10.0 (12-15 X 9-11) micron; a compact sporocyst residuum was present, but Stieda, sub-, and parastieda bodies were absent. This species was found in 2 of 18 (11%) hosts. Sporulated oocysts of Eimeria neurotrichi n. sp. were ovoid, 17.6 X 13.6 (16-20 X 11-16) micron; micropyle and oocyst residuum were absent, but a polar body was present. Ovoid sporocysts were 10.7 X 5.5 (9-12 X 5-6) micron; Stieda body and sporocyst residuum were present, but sub- and parastieda bodies were absent. This species was found in 2 of 18 (11%) hosts. Sporulated oocysts of Eimeria parastiedica n. sp. were subspheroid, 27.4 X 25.5 (25-30 X 22-28) micron; micropyle, oocyst residuum, and polar body were absent. Ovoid sporocysts, pointed at both ends, were 18.3 X 10.4 (16-20 X 9-11) micron; Stieda, sub-, and parastieda bodies were present as was a sporocyst residuum. This species was found in 2 of 18 (11%) hosts.(ABSTRACT TRUNCATED AT 250 WORDS)

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Invasion and early development of Sarcocystis muris (Apicomplexa, Sarcocystidae) in tissue cultures.

Ultrastructural observations on the invasion and early development of merozoites (bradyzoites) of Sarcocystis muris in Madin-Darby canine kidney (MDCK) cells are presented. Invading merozoites cause the host cell plasmalemma to invaginate; they form a membrane junction (moving junction) and move into the host cell where they are enclosed in a primary parasitophorous vacuole (PV). Within 30-45 min after becoming intracellular, merozoites begin to vacate the newly established primary PV and move, forming a new membrane junction, into a secondary PV. Simultaneously with the movement of the parasite, the contents of dense granules in the apical part of the merozoites are shed by exocytosis into the lumen of the developing secondary PV. A lamella of the endoplasmic reticulum of the host cell becomes attached to the PV membrane, forming a PV limited by three host cell membranes.

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Eimeria ladronensis n. sp. and E. albigulae (Apicomplexa: Eimeriidae) from the woodrat, Neotoma albigula (Rodentia: Cricetidae).

Of 50 white-throated woodrats (Neotoma albigula) collected from Socorro Co., New Mexico, 21 (42%) had eimerian oocysts in their feces when examined. Of the 21 Neotoma found positive for Eimeria, 19 (90%) harbored a single eimerian species at time of examination. Eimeria albigulae Levine, Ivens & Kruidenier, 1957, was found in 18 (86%), and E. ladronensis n. sp. was found in five (24%) infected woodrats. Sporulated oocysts of E. ladronensis are ellipsoidal, 19-25 X 13-15 (21.4 +/- 1.3 X 14.1 +/- 1.1) micron, have a smooth wall and one or two polar granules, but lack a micropyle and an oocyst residuum. Sporocysts are tapered at one end, 7-10 X 6-7 (8.5 +/- 0.7 X 6.5 +/- 0.3) micron, and have a Stieda body and sporocyst residuum, but no substieda body. Prepatent periods for E. albigulae and E. ladronensis n. sp. are 5-6 and 8-9 days, respectively; patent periods are 7-18 and approximately 11 days, respectively.

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Coccidian parasites (Apicomplexa: Eimeriidae) from insectivores. II. Six new species from Japanese shrew moles (Talpidae).

Thirty-eight of 51 (74.5%) shrew moles collected in Japan were infected with from one to four species of Eimeria and/or Isospora including six of six Dymecodon pilirostris and 32 of 45 (71.1%) Urotrichus talpoides. Four eimerians and two isosporans were identified and all are described as new species. Sporulated oocysts of Eimeria amorphospora n. sp. were subspheroid/ellipsoid, 21.1 x 17.9 (18-25 x 16-21) micrometers. Sporocysts were amorphous, gelatinoid envelopes greater than or equal to 20.3 x 7.5 (17-24 x 7-9) micrometers. Sporozoites were enclosed together within a membrane in each sporocyst. This species was found in 9 of 45 (20%) U. talpoides. Sporulated oocysts of Eimeria gonocilia n. sp. were subspheroid/ellipsoid, 28.8 x 24.4 (25-30 x 21-28) micrometers; a highly ornate outer oocyst wall gave the appearance of a ciliated ball. Sporocysts ovoid, pointed at both ends, were 17.0 x 9.9 (15-19 x 7-11) micrometers; this species was found in 4 of 45 (8.9%) U. talpoides. Sporulated oocysts of Eimeria talpoidei n. sp. were asymmetrical ovoid, 20.6 x 13.3 (18-23 x 12-15) micrometers, with sporocysts lacrimiform, 12.0 x 5.8 (10-14 x 5-7) micrometers. This species was found in 7 of 45 (15.6%) U. talpoides. Sporulated oocysts of Eimeria honshuensis n. sp. were ellipsoid, 15.5 x 11.4 (13-18 x 10-13) micrometers, with sporocysts ovoid, 9.1 x 5.2 (8-10 x 4-6) micrometers. This species was found in 10 of 45 (22.2%) U. talpoides and in 5 of 6 (83.3%) D. pilirostris. sporulated oocysts of Isospora dymecodi n. sp. were subspheroid/ellipsoid, 15.8 x 12.6 (13-17 x 11-13) micrometers, with sporocysts ovoid, 9.1 x 5.2 (8-10 x 4-6) micrometers. This species was found in 10 of 45 (22.2%) U. talpoides and in 5 of 6 (83.3%) D. pilirostris. Sporulated oocysts of Isospora dymecodi n. sp. were subspheroid/ellipsoid, 15.8 x 12.6 (13-17 x 11-13) micrometers, with sporocysts ellipsoid, 10.9 x 6.9 (10-13 x 6-8). This species was found in six of six D. pilirostris. Sporulated oocysts of Isospora urotrichi n. sp. were spheroid/subspheroid, 13.4 x 12.4 (11-16 x 9-14) micrometers, with sporocysts ovoid, 9.2 x 6.3 (8-11 x 5-7) micrometers. This species was found in 27 of 45 (60%) U. Talpoides. Only 14 of 38 (36.8%) infected hosts (one D. pilirostris, 13 U. talpoides) were seen to be naturally infected with only one coccidian species when sampled.

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Coccidia (Apicomplexa: Eimeriidae) from sciurid rodents (Eutamias, Sciurus, Tamiasciurus spp.) from the western United States and northern Mexico with description of two new species.

Since May 1979, 190 rodents in the family Sciuridae, representing three genera and nine species, have been collected in the western United States and northern Mexico and examined for coccidia; 71 (37%) had coccidian oocysts in their feces. These included 2 of 12 (17%) Eutamias canipes; 7 of 12 (58%) E. dorsalis; 18 of 50 (36%) E. merriami; 33 of 96 (34%) E. obscurus; 3 of 4 (75%) E. townsendii; 3 of 9 (33%) Sciurus aberti; 1 of 1 S. griseus; 1 of 1 Tamiasciurus hudsonicus mogollonensis; and 3 of 5 (60%) T. mearnsi. The following coccidians were identified from infected rodents: Eimeria cochisensis n. sp. and Eimeria dorsalis n. sp. from E. canipes, E. cochisensis, E. dorsalis, and E. tamiasciuri from E. dorsalis, E. dorsalis and E. tamiasciuri from E. merriami; E. cochisensis, E. dorsalis, E. tamiasciuri, and E. wisconsinensis from E. obscurus; E. cochisensis and E. dorsalis from E. townsendii; E. ontarioensis and E. tamiasciuri from S. aberti; E. tamiasciuri from S. griseus; E. tamiasciuri and E. toddi from T. h. mogollonensis; and E. tamiasciuri from T. mearnsi. Sporulated oocysts of Eimeria dorsalis n. sp. were ovoid, 21.9 x 16.8 (17-24 x 14-20) micrometer with sporocysts ovoid, 11.5 x 6.9 (10-14 x 6-8) micrometer. Sporulated oocysts of Eimeria cochisensis n. sp. were spheroid to subspheroid, 16.7 x 15.3 (15-18 x 14-17) micrometer, with sporocysts ovoid, 8.4 x 5.6 (6-11 x 4-7) micrometer. Fifty-five of 71 (77%) infected hosts had oocysts of only one eimerian species in their feces at the time they were examined. One eimerian, E. tamiasciuri, was found in seven of nine host species in three genera. A list is provided of all eimerians (22, including the species described here) that have been described in the literature from Eutamias, Sciurus, and Tamiasciurus spp.

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The life cycle of Cryptosporidium baileyi n. sp. (Apicomplexa, Cryptosporidiidae) infecting chickens.

The life cycle and morphology of a previously undescribed species of Cryptosporidium isolated from commercial broiler chickens is described. The prepatent period for Cryptosporidium baileyi n. sp. was three days post oral inoculation (PI) of oocysts, and the patent period was days 4-24 PI for chickens inoculated at two days of age and days 4-14 for chickens inoculated at one and six months of age. During the first three days PI, most developmental stages of C. baileyi were found in the microvillous region of enterocytes of the ileum and large intestine. By day 4 PI, most parasites occurred in enterocytes of the cloaca and bursa of Fabricius (BF). Mature Type I meronts with eight merozoites first appeared 12 h PI and measured 5.0 x 4.9 micrometers. Mature Type II meronts with four merozoites and a large granular residuum first appeared 48 h PI and measured 5.1 x 5.1 micrometers. Type III meronts with eight short merozoites and a large homogeneous residuum first appeared 72 h PI and measured 5.2 x 5.1 micrometers. Microgamonts (4.0 x 4.0 micrometers) produced approximately 16 microgametes that penetrated into macrogametes (4.7 x 4.7 micrometers). Macrogametes gave rise to two types of oocysts that sporulated within the host cells. Most were thick-walled oocysts (6.3 x 5.2 micrometers), the resistant forms that passed unaltered in the feces. Some were thin-walled oocysts whose wall (membrane) readily ruptured upon release from the host cell. Sporozoites from thin-walled oocysts were observed penetrating enterocytes in mucosal smears. The presence of thin-walled, autoinfective oocysts and the recycling of Type I meronts may explain why chickens develop heavy intestinal infections lasting up to 21 days. Oocysts of C. baileyi were inoculated orally into several animals to determine its host specificity. Cryptosporidium baileyi did not produce infections in suckling mice and goats or in two-day-old or two-week-old quail. One of six 10-day-old turkeys had small numbers of asexual stages only in the BF. Four of six one-day-old turkeys developed mild infections only in the BF, and sexual stages of the parasite were observed in only one of the four. All seven one-day-old ducks and seven two-day-old geese developed heavy infections only in the BF with all known developmental stages present.

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Isospora elmahalensis n. sp. (Apicomplexa, Eimeriidae), a parasite of the white-cheeked bulbul (Pycnonotus leucogenys) in Saudi Arabia.

Isospora elmahalensis n. sp. is described from the Saudi Arabian bird, Pycnonotus leucogenys, from the Elmahala valley. Sporulated oocysts of I. elmahalensis were spherical or nearly subspherical, 19.5-22.5 X 18.5-20 (21.34 +/- 0.4 X 19.06 +/- 0.5) microns. Oocysts lacked a micropyle, residuum, and polar granule. Sporocysts were ovoid, 14-17.5 X 7-12 (16.08 +/- 1.05 X 9.9 +/- 1.55) microns, and had a Stieda body and sporocyst residuum, but lacked a substiedal body. Sporozoites were elongated with a clear globule at one end. The host bird belongs to the order Passeriformes.

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Corrections in the names of rodent coccidia (Apicomplexa, Coccidiasina).

The following new taxonomic combinations are introduced for coccidia whose names were previously given erroneously: Dorisa bengalensis (Bandyopadhyay & Ray, 1982) n. comb. from the Indian palm squirrel Funambulus pennanti in India; Eimeria sicistae from the intestine of the birch mouse Sicista tianschanica in the USSR; E. hydrochaeri Carini, 1937 emend. from the capybara Hydrochaerus hydrochaerus in South America; Frenkelia sp. (Doby, Jeannes & Rault 1965) from the brain of the water vole Arvicola sapidus in Europe; Frenkelia sp. (Karstad, 1963) from the brain of the muskrat Ondatra zibethica in North America; Frenkelia sp. (Enemar, 1965) from the brain of the lemming Lemmus lemmus in Europe; Frenkelia sp. (Sebek, 1975) from the brain of the field mouse Apodemus flavicollis in Europe; and Sarcocystis sp. (Ryan, Wyand & Nielsen, 1982) from the skeletal muscles of the muskrat Ondatra zibethica in North America.

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Eimeria tahamensis N. sp. (Apicomplexa: Eimeriidae) from the Arabian quail (Coturnix delegorguei arabica).

Eimeria tahamensis n. sp. is described from the harlequin quail (Coturnix delegorguei arabica) from Tahama, Saudi Arabia. The sporulated oocysts of E. tahamensis n. sp. are ellipsoid, 36.5-42 X 25.5-29 (41.2 +/- 1.34 X 28.4 +/- 0.81) micron, with a thick two-layered wall and one polar granule but without a micropyle or an oocyst residuum. The sporocysts are ovoid, 14-16 X 9-11.5 (15.3 +/- 0.7 X 10.8 +/- 0.64) micron, with a knob-like Stieda body and sporocyst residuum, but without a substiedal body. The sporozoites are often located transversely at the two ends of the sporocysts. The host bird belongs to the order Galliformes.

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Caryocyst-like host cell formation by Caryospora duszynskii (Apicomplexa: Eimeriidae) in human fetal lung cell cultures.

Sporozoites of the coccidium, Caryospora duszynskii, penetrated human fetal lung cell cultures but did not undergo asexual or sexual multiplication during a 29-day observation period. Beginning three days postinoculation (PI), infected host cells lost their normal elongated fibroblast-like shape and became ellipsoidal in appearance and resembled caryocysts. These caryocyst-like infected cells were observed from 3 through 29 days PI. Sporozoites remained viable throughout the study as evidenced by motility of extracellular sporozoites in infected human fetal lung cell cultures. Results of this in vitro study suggest that some species of Caryospora may form caryocysts in secondary hosts without undergoing asexual or sexual multiplication in these hosts.

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Coccidian parasites (Apicomplexa: Eimeriidae) from insectivores. V. Ten forms from the moles of Japan (Euroscaptor, Mogera spp.).

Moles from Japan were examined for coccidian oocysts, and 67 of 77 (87%) hosts were infected including 8 of 11 (73%) Euroscaptor mizura, 31 of 36 (86%) Mogera kobeae, 17 of 17 M. tokudae, and 11 of 13 (85%) M. wogura. Of 67 infected hosts, 57 (85%) had multiple infections representing 2-5 coccidian species when examined. All oocysts in the infected fecal samples remained unsporulated and the absence of sporulation may be the result of storing feces from Japanese moles in 2% aqueous H2SO4. Five structurally distinct forms of unsporulated oocysts were found in E. mizura, and five distinct forms of unsporulated oocysts were also seen in Mogera spp. Two of the forms from E. mizura were similar to forms from Mogera spp., and the five forms from Mogera were shared freely between the three Mogera species. This is the first systematic survey of Japanese moles for coccidia.

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Coccidian parasites (Apicomplexa: Eimeriidae) from insectivores. IV. Four new species in Talpa europaea from England.

Moles from England were examined for coccidian oocysts and all 64 Talpa europaea were infected; of 64 infected hosts, 56 (88%) had multiple infections representing two to six coccidian species when examined. Oocysts in 31 of the 64 samples remainedunsporulated. Three eimerians and one isosporan were studied from the 33 fecal samples that had sporulated oocysts and these are described as new species; Cyclospora talpae Pellérdy & Tanyi, 1968, and Isospora sofiae (Golemansky, 1978) Levine & Ivens, 1979, are redescribed; and Cyclospora sp., similar to C. talpae, is discussed. Sporulated oocysts of C. talpae are ellipsoidal, 14.3 X 9.6 (12-19 X 6-13) microns with sporocysts ovoid, 9.4 X 5.7 (6-13 X 4-8) microns; it was found in 21 of the 33 (63.6%) sporulated samples. Sporulated oocysts of Cyclospora sp. are subspheroidal to ellipsoidal, 12.5 X 8.9 (10-14 X 6-12) microns with sporocysts ovoid, 8.6 X 5.3 (6-10 X 4-6) microns; it was found in 21 of the 33 (63.6%) sporulated samples. Sporulated oocysts of Eimeria avonensis n. sp. are elongate-ellipsoidal, 15.0 X 9.6 (13-20 X 7-12) microns with sporocysts ovoid, 6.6 X 3.6 (5-9 X 3-7) microns; it was found in 15 of the 33 (45.5%) sporulated samples. Sporulated oocysts of Eimeria berea n. sp. are subspheroidal, 12.1 X 10.5 (10-15 X 8-14) microns with sporocysts ovoid, 6.3 X 3.9 (5-10 X 2-5) microns; it was found in 8 of the 33 (24.2%) sporulated samples.(ABSTRACT TRUNCATED AT 250 WORDS)

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Prevalence of Eimeria spp. (Apicomplexa: Eimeriidae) in Oregon llamas.

Eimeria alpacae, E. punoensis, E. lamae, and E. macusaniensis were identified in fecal samples from 189 llama (Lama glama (L.] adults and 50 llama crias (animals less than one year of age of any species in the genus Lama) from central and western Oregon. In both adults and crias, E. alpacae was the most common species found. The least common was E. macusaniensis, which was found in only two adults. Overall prevalence and numbers of animals with mixed infections was approximately twice as high in crias as in adults.

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Eimeria sinaitae n. sp. (Apicomplexa: Eimeriidae) from the rock agama (Agama sinaita) in Saudi Arabia.

Eimeria sinaitae n. sp. is described from the gall bladder of Agama sinaita from Wasie, Saudi Arabia. Sporulated oocysts are elongate-ellipsoid 34.4 x 22.0 (29.0-40.0 x 17.4-24.5) micron. Oocyst wall is smooth, greenish yellow, 1.2 (1.0-1.4) micron thick, and two-layered. Micropyle, polar granule, and oocyst residuum are absent. Sporocysts are ellipsoid 11.4 x 7.6 (9.8-15.0 x 6.7-9.0) micron. Sporocyst residuum is present. The sporocysts lack a Stieda body. Sporozoites are crescent-shaped, blunt at one end and tapered at the other. Eimeria species from Agamidae are compared.

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Cross-transmission of Eimeria spp. (Protozoa, Apicomplexa) of rodents--a review.

A total of 169 cross-transmission attempts has been made with 44 (11.8%) of the 372 named species of Eimeria of rodents. Of these, 161 were rodent-to-rodent, 6 rodent-to-lagomorph, and 1 each rodent-to-carnivore and rodent-to-bird. None of the last three categories was successful. In the rodent-to-rodent combinations, 39 (80%) of the 49 attempts to transmit a coccidian species from one rodent species to another of the same genus were successful, and only 14 (12.5%) of the 112 attempts to transmit a coccidium to a rodent of a different genus were successful. Eight of the successful attempts were with E. chinchillae, which was the only truly euryxenous species of Eimeria in the group. Two successful attempts were between between the closely related rodent genera Spermophilus and Cynomys, and two were both of E. separata from Rattus norvegicus to some genetic strains but not to others of Mus musculus. One attempt with E. vermiformis from Mus musculus to Rattus norvegicus required treatment of the rat with the immunosuppressant dexamethasone to succeed. More cross-transmission studies are needed to determine the host spectra of the species of Eimeria and other coccidian genera, and to determine the roles of genetics and immunosuppression in their transmission.

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Complete development of Caryospora bigenetica (Apicomplexa: Eimeriidae) in vitro.

The development of Caryospora bigenetica in vitro is described by light microscopy. Sporozoites from snake-derived oocysts were purified and inoculated onto cultures of primary testicle cells of the cotton rat, cotton rat kidney cells, and human fetal lung cells. Intracellular sporozoites were observed one and two days postinoculation (DPI). Motile, extracellular first-generation merozoites were present 3 DPI, and second-generation merozoites were present 5 DPI. Mature gamonts were observed 9 DPI and developed into unsporulated oocysts by 10 DPI. Oocystes sporulated in vitro, and excystation was observed. Cells that were penetrated by in vitro-produced sporozoites formed caryocysts by 16 DPI. To test infectivity of in vitro-derived stages, merozoites were removed from cultured cells 5 DPI and inoculated intraperitoneally into a mouse; infection resulted. Sporulated oocysts removed from cell cultures 12 DPI produced facial swelling in an orally inoculated cotton rat.

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