In Vitro Excystation of Caryospora simplex (Apicomplexa: Eimeriorina)1

In vitro excystation of sporozoites of the heteroxenous coccidian Caryospora simplex Léger, 1904 (Apicomplexa: Eimeriorina) is described. Sporocysts freed mechanically from oocysts released a maximum of 51% of their sporozoites within 45 min at 25°C and a maximum of 74% within 20 min at 37°C when incubated in a 0.25% (w/v) trypsin–0.75% (w/v) sodium taurocholate (bile salt) excystation solution. At emergence from sporocysts, sporozoites were weakly motile then became highly active after about 2 min in excystation solution. Sporozoites within sporocysts exposed to bile salt only became highly motile within 25 min at 25°C and within 15 min at 37°C but did not excyst. When exposed only to trypsin at the above temperatures, the Stieda body dissolved; the substieda body remained intact, and the sporozoites exhibited only limited motility within sporocysts; only a few excysted. Intact, sporulated oocysts incubated at 25° or 37°C in 0.02 M cysteine-HC1 and a 50% CO₂ atmosphere for 18 h had no morphologic changes in the oocyst wall. Further incubation of these intact oocysts in excystation solution for 30 min at 37°C caused neither motility of sporozoites within sporocysts nor excystation. Grinding oocysts for 30 sec in a motor-driven, teflon-coated tissue grinder caused motility of some sporozoites within sporocysts but did not result in excystation.     

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.     

A redescription of Cryptosporidium galli Pavlasek, 1999 (Apicomplexa: Cryptosporidiidae) from birds

Cryptosporidium galli Pavlasek, 1999, described from the feces of birds, is redescribed with additional molecular and biological data. Oocysts are ellipsoidal, are passed fully sporulated, lack sporocysts, and measure 8.25 x 6.3 microm (range 8.0-8.5 x 6.2-6.4 microm) with a length-width ratio of 1.30 (n = 50). Oocysts are structurally similar to those of Cryptosporidium baileyi described from chickens, but in addition to being considerably larger than oocysts of C. baileyi, these oocysts infect the proventriculus in a variety of birds and not the respiratory tract. Oocysts were successfully transmitted from chickens to chickens, and morphologically similar oocysts also were observed in a variety of exotic and wild birds (Order Passeriformes, Phasianidae, Fringillidae, and Icteridae). Molecular and phylogenetic analyses at the 18S rRNA, HSP70, and actin gene loci demonstrate that this species is genetically distinct from all known species and genotypes of Cryptosporidium and, thus, was named C. galli.     

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)     

Clinical large intestinal coccidiosis in camels (Camelus dromedarius) in the United Arab Emirates: description of lesions,endogenous stages,and redescription of Isospora orlovi,Tsygankov, 1950 oocysts

Between January and March 2001, eight 4- to 8-wk-old camels (Camelus dromedarius) from 2 farms from Dubai area of the United Arab Emirates were submitted for necropsy examination. The camels had diarrhea of 2-5 days duration. Grossly, a severe diphtheroid-to-hemorrhagic colitis was seen in all animals. Gamonts, unsporulated oocysts, sporulating oocysts, and fully sporulated oocysts were present in the intestinal epithelium and the lamina propria. Fully sporulated oocysts contained 2 sporocysts and 4 sporozoites in each sporocyst. Oocysts from fecal samples resembled oocysts of Isospora orlovi. This is the first report of an isosporan parasite associated with hemorrhagic enteritis in the large intestine of any animal.     

Enzyme Variation of Eimeria arizonensis from Peromyscus truei and P. boylii

ABSTRACT. Cricetid rodents, Peromyscus truei and P. boylii , were inoculated with sporulated oocysts of Eimeria arizonensis collected from wild P. truei maintained in the lab. In P. truei the prepatent period was 4–5 days, the patent period was 9–11 days, and sporulated oocysts were 21.5 × 25.0 (20–23 × 24–26) μm with sporocysts 7.7 × 12.0 (6–8 × 10–13) pm. In P. boylii the prepatent period was 6–7 days, the patent period was 8–9 days, and sporulated oocysts were 20.1 × 23.2 (18–22 × 21–24) pm with sporocysts 6.8 × 10.0 (5–8 × 9–12) pm. Sporulated oocysts from both host species were used in direct side-by-side comparison of isozyme banding patterns using protein electrophoresis. The parasite has polytypic loci for leucine aminopeptidase (LAP), lactate dehydrogenase (LDH), and 6-phosphogluconate dehydrogenase (6-PGD). In oocysts from P. truei , LAP showed one band with fast migration and LDH and 6-PGD each showed two bands, one with fast and one with slow migration. In oocysts from P. boylii , LAP and LDH each had one band with slow migration and 6-PGD had one band with moderate migration. Oocysts of E. arizonensis collected from P. boylii were used to inoculate P. truei. The prepatent and patent periods, structural measurements, and isozyrne banding patterns of the resultant oocysts were the same as those from P. truei when inoculated with oocysts from P. truei.     

Coccidian Parasites (Apicomplexa: Eimeriidae) from Insectivores. IV. Four New Species in Talpa europaea from England1

ABSTRACT. 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 remained unsporulated. 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 × 9.6 (12–19 × 6–13) μm with sporocysts ovoid, 9.4 × 5.7 (6–13 × 4–8) μm; it was found in 21 of the 33 (63.6%) sporulated samples. Sporulated oocysts of Cyclospora sp. are subspheroidal to ellipsoidal, 12.5 × 8.9 (10–14 × 6–12) μm with sporocysts ovoid, 8.6 × 5.3 (6–10 × 4–6) μm; it was found in 21 of the 33 (63.6%) sporulated samples. Sporulated oocysts of Eimeria avonensis n. sp. are elongate-ellipsoidal, 15.0 × 9.6 (13–20 × 7–12) μm with sporocysts ovoid, 6.6 × 3.6 (5–9 × 3–7) μm; it was found in 15 of the 33 (45.5%) sporulated samples. Sporulated oocysts of Eimeria berea n. sp. are subspheroidal, 12.1 × 10.5 (10–15 × 8–14) μm with sporocysts ovoid, 6.3 × 3.9 (5–10 × 2–5) μm; it was found in 8 of the 33 (24.2%) sporulated samples. Sporulated oocysts of Eimeria globula n. sp. are spheroidal, 20.9 × 19.9 (19–24 × 17–21) μm with sporocysts elongate-ovoid, 11.5 × 6.9 (9–16 × 6–9) μm; it was found in 3 of the 33 (9.1%) sporulated samples. Sporulated oocysts of Isospora sporopointaea n. sp. are subellipsoidal to ellipsoidal, 17.1 × 11.4 (13–21 × 8–14) μm with sporocysts ellipsoidal with both ends pointed, 11.9 × 5.9 (9–16 × 4–8) μm; it was found in 27 of the 33 (81.8%) sporulated samples. Sporulated oocysts of I. sofiae are spheroidal to subspheroidal, 12.2 × 11.0 (9–16 × 8–15) μm with sporocysts ovoid, 9.1 × 5.2 (6–13 × 3–8) μm; it was found in 25 of the 33 (75.8%) sporulated samples. To date, the coccidian parasites of talpids include two cyclosporans, 12 eimerians, and six isosporans, exclusive of the four new species described here.     

Enzyme variation of Eimeria arizonensis from Peromyscus truei and P. boylii

Cricetid rodents, Peromyscus truei and P. boylii, were inoculated with sporulated oocysts of Eimeria arizonensis collected from wild P. truei maintained in the lab. In P. truei the prepatent period was 4-5 days, the patent period was 9-11 days, and sporulated oocysts were 21.5 x 25.0 (20-23 x 24-26) microns with sporocysts 7.7 x 12.0 (6-8 x 10-13) microns. In P. boylii the prepatent period was 6-7 days, the patent period was 8-9 days, and sporulated oocysts were 20.1 x 23.2 (18-22 x 21-24) microns with sporocysts 6.8 x 10.0 (5-8 x 9-12) microns. Sporulated oocysts from both host species were used in direct side-by-side comparison of isozyme banding patterns using protein electrophoresis. The parasite has polytypic loci for leucine aminopeptidase (LAP), lactate dehydrogenase (LDH), and 6-phosphogluconate dehydrogenase (6-PGD). In oocysts from P. truei, LAP showed one band with fast migration and LDH and 6-PGD each showed two bands, one with fast and one with slow migration. In oocysts from P. boylii, LAP and LDH each had one band with slow migration and 6-PGD had one band with moderate migration. Oocysts of E. arizonensis collected from P. boylii were used to inoculate P. truei. The prepatent and patent periods, structural measurements, and isozyme banding patterns of the resultant oocysts were the same as those from P. truei when inoculated with oocysts from P. truei.     

Cryptosporidium fayeri n. sp. (Apicomplexa: Cryptosporidiidae) from the Red Kangaroo (Macropus rufus)

The morphology and infectivity of the oocysts of a new species of Cryptosporidium from the faeces of the red kangaroo (Macropus rufus) are described. Oocysts are structurally indistinguishable from those of Cryptosporidium parvum. Oocysts of the new species are passed fully sporulated, lack sporocysts, and measure 4.5-5.1 microm (mean=4.9) x 3.8-5.0 microm (mean=4.3 microm) with a length to width ratio 1.02:1.18 (mean 1.14) (n=50). Oocysts were not infectious for neonate ARC Swiss mice. Multi-locus analysis of numerous unlinked loci demonstrated this species to be distinct (90.64%-97.88% similarity) from C. parvum. Based on biological and molecular data, this Cryptosporidium infecting marsupials is proposed to be a new species Cryptosporidium fayeri n. sp.     

Excystation of the Poultry Coccidium, Eimeria acervulina

SYNOPSIS. Using intervals up to 5 hours, attempts to excyst sporozoites of Eimeria acervulina from intact oocysts in vitro were unsuccessful.
Examination of crop, gizzard, and intestinal contents of chicks fed large numbers of sporulated oocysts indicated that (1) no obvious change in the oocysts occurred in the crop, (2) a high percentage of the sporocysts were quickly released from the oocysts in the gizzard, (3) the sporozoites escaped from the liberated sporocysts in the duodenum and jejunum, and (4) the action of the digestive juice was apparently on the sporocysts rather than on the oocysts.
In vitro attempts to excyst sporozoites from free sporocysts with various pancreatic preparations in the absence of bile produced low or insignificant percentages of excystation. In the presence of bile, bile salts, and other surface-active agents, the action of the pancreatic preparations was greatly increased. The heaviest suspension of motile, nonaggregating sporozoites was obtained with 0.25% trypsin 1–300 in 5% chicken bile at pH 7.6.     

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.     

Caryospora cobrae n. sp. from the Indian Cobra Naja naja Linnaeus

Oocysts of Caryospora cobrae n. sp. were isolated from an Indian cobra, Naja naja Linnaeus from West Bengal, India. The sporulated oocysts were spherical to subspherical, 16.5-19.5 times 16.5-18.0 μm, and had a micropyle but lacked a polar granule and oocysl residuum. The sporocysts were piriform, measuring 12.0-16.5 times 9.0?12.8 μm; a Stieda body and sporocyst residuum were present.     

Effect of the anticoccidial arprinocid on production, sporulation, and infectivity of Eimeria oocysts

Medication of broilers with arprinocid [MK-302, 9-(2-chloro-6-fluorbenzyl adenine)] had 3 distinct effects on oocysts; (1) the number of oocysts produced was decreased, (2) fewer of the oocysts sporulated, and (3) those oocysts which did sporulate were less infective than those from unmedicated birds. The drug level necessary to prevent passage of oocysts depended on the species and strain of coccidia. To essentially eliminate oocyst production (less than 5% of controls) required medication with the following levels of arprinocid: 70 ppm with Eimeria maxima; 60 ppm with E. mivati, E. E. necatrix, and E. brunetti; and 50 ppm with E. tenella. With E. acervulina, oocysts were completely eliminated by 60 ppm of arprinocid with one field strain but were still numerous at 70 ppm with a second field strain. Oocysts recovered from birds on medication often failed to sporulate. No sporulation was seen at drug levels of 30 ppm or above with E. maxima and E. mivati. The level of arpinocid required to prevent sporulation with other species depended on the strain being studied, but varied from 30 ppm to 70 ppm. The oocysts of E. acervulina, E. mivati, E. tenella, and E. brunetti recovered from medicated birds that subsequently sporulated, were less infective when inoculated into susceptible birds, than oocysts from unmedicated birds. Oocysts from low medication level with E. necatrix (30 ppm) and E. maxima (10 ppm), once sporulated, were as infective as oocysts from unmedicated control birds, even though the numbers produced were less. No differences were detected in the time oocysts were produced between medicated and unmedicated birds infected with E. acervulina, E. maxima, E. brunetti, and E. tenella.     

Caryospora uptoni and Frenkelia sp.-like coccidial infections in red-tailed hawks (Buteo borealis)

The feces from 16 red-tailed hawks (Buteo borealis) were examined by fecal flotation for the presence of coccidial oocysts or sporocysts. Oocysts of Caryospora uptoni were found in five (31%), sporocysts of a Frenkelia sp.-like coccidium were found in eight (50%), and mixed infections with both species of coccidia were found in three (19%) red-tailed hawks. Neither oocysts nor sporocysts were found in six (38%) red-tailed hawks. Sexual stages of C. uptoni were found in the duodenum and jejunum of a naturally infected red-tailed hawk. Sexual stages were located in enterocytes on the distal three-fourths of the villi. This study shows that over 60% of red-tailed hawks may be passing coccidial oocysts/sporocysts in their feces and provides morphological information for diagnosing C. uptoni infections in histological sections.     

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.     

Descriptions of two new species of coccidia (Protozoa: Eimeriidae) and redescriptions of Eimeria ivensae and Eimeria odocoilei from captive white-tailed deer, Odocoileus virginianus

Two new species of Eimeria were observed in the feces of captive white-tailed deer fawns, Odocoileus virginianus, from Alabama. The first new species was easily recognized because of its small size. Sporulated oocysts are spherical, average 10.2 by 10.0 microm, and lack a micropyle and oocyst residuum. Oocysts contain a polar granule and elongate-ellipsoidal sporocysts that measure 6.7 by 3.1 microm. A Stieda body is present on the sporocysts. Oocysts were observed in the feces, and gamonts and oocysts were observed in the jejunum of a month-old fawn from Minnesota that died from enteritis due to this species. Oocysts of this small species were present in 5 of the 6 white-tailed deer fawns examined. Oocysts of a second new species are ellipsoidal and average 29.5 by 24.6 microm. The oocyst encloses an oocyst residuum, polar granule, and elongate-ellipsoidal sporocysts that average 16.0 by 9.0 microm. A Stieda body and substieda body are present on the sporocysts. Oocysts of the second new species were present in 4 of the 6 white-tailed deer fawns examined. Oocysts of E. ivensae are ovoid or flask-like and average 32.0 by 20.8 microm. The oocyst wall is rough, contains a micropyle, and encloses elongate-ellipsoidal sporocysts that average 16.5 by 7.8 microm. A Stieda body is present on the sporocysts. Oocysts of E. ivensae were present in 4 of the 6 white-tailed deer fawns. Oocysts of E. odocoilei are spherical or slightly subspherical and measure 24.7 by 21.5 microm. They enclose ovoid sporocysts that average 12.7 by 8.8 microm. A Stieda and substieda body are present on the sporocyst. Oocysts of E. odocoilei were present in 4 of the 6 white-tailed deer fawns.     

Eimeria and sarcocystis in raccoons in Illinois

Eimeria nuttalli oocysts were found in 58% (21/36) and E. procyonis oocysts in 25% (9/36) of raccoons Procyon lotor in Illinois, and sporocysts of Sarcocystis sp. in 17% (2/12) of other raccoons in Illinois. The oocysts of E. nuttalli were ellipsoidal to ovoid, 15-21 X 12-17 micrometer, with a one-layered, smooth, colorless wall. The oocysts of E. procyonis were 22-28 X 18-22 micrometer, with a rough, striated, brownish, two-layered wall. The sporulated sporocysts of Sarcocystis sp. were 11-13 X 8-10 micrometer. Attempts to infect baby pigs by feeding them sporocysts of Sarcocystis sp. from the reaction failed.     

A new species of Cryptosporidium (Apicomplexa: Cryptosporidiidae) from eastern grey kangaroos (Macropus giganteus)

Cryptosporidium macropodum n. sp is described. Oocysts of C. macropodum from the feces of kangaroos (Macropus spp.) are morphologically indistinguishable from other mammalian Cryptosporidium species, including C. parvum, C. hominis, C. suis, and C. canis. The oocysts are fully sporulated on excretion, lack sporocysts, and have an average width of 4.9 microm (4.5-6.0), a length of 5.4 microm (5.0-6.0), and a length:width ratio of 1.1. Phylogenetic analyses of the 18S ribosomal RNA, actin, and heat shock protein 70 (HSP70) loci demonstrate that C. macropodum is genetically distinct from all described Cryptosporidium species, including others found in marsupials. The parasite seems to be highly host-specific, because it has been found only in marsupials to date. Therefore, based on biological and molecular data, we consider C. macropodum a new species.     

Eimeria malabaricas sp. n. and Eimeria bandipurensis from the South Indian Tree Squirrel,Funambulus tristriatus*

Synopsis. Oocysts of Eimeria malabaricas sp. n. and Eimeria bandipurensis from the South Indian tree squirrel Funambulus tristriatus collected in Kerala, India, are described. Sporulated oocysts of E. malabaricas were ellipsoid to subspherical measuring 39.8 (35–45) × 32.1 (29–37) m?m, with a thick (2.5–3.0 m?m), 2-layered wall. The outer layer was yellow-brown, striated, and rough. There was no micropyle. but a polar granule was present in 34% of oocysts. The sporocysts were ovoid, 16.0 (14.0–18.0) × 11.2 (11.0–12.0) m?m, with a Stieda body and a granular residuum. Excysted sporozoites were 21.8 (19.0–23.0) × 3.4 (3.0–4.0) m?m, with a large refractile body. The sporulated oocysts of E. bandipurensis are redescribed.     

Additional Notes on the Coccidian Parasites of the Soft-shell Turtle,Trionyx spiniferus Le Sueur,in Iowa,with a Description of Eimeria vesicostieda sp. n.*

SYNOPSIS Oocysts of Eimeria dericksoni Roudabush, Eimeria mascoutini Wacha & Christiansen, and Eimeria vesicostieda sp. n. were isolated from the Spiny Soft-shell Turtle, Trionyx spiniferus LeSueur, in Iowa. The sporulated oocysts of E. dericksoni are redescribed to include, for the first time, the dimensions of the sporocysts (6.0–8.5 × 3.0–5.0 μm) and the structural appearance of the Stieda body (thinly convex); the oocysts of E. vesicostieda are newly described as being thickwalled, narrowly ovoid to narrowly ellipsoid, measuring 22.0–25.5 × 16.5–20.5 μm. and having a polar granule, a sporocyst residuum, and a vesicle-like Stieda body; the oocysts of E. mascoutini are documented photographically.