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PubMed1988

Coccidian parasites (Apicomplexa: Eimeriidae) from insectivores. VI. Six new species from the eastern mole, Scalopus aquaticus.

Thirteen eastern moles, Scalopus aquaticus, collected in West Texas were examined for coccidian oocysts; 11 (85%) were infected and eight (73%) of these had multiple infections representing two or more species. One cyclosporan, three eimerians, and two isosporans were studied and all are described as new species. Sporulated oocysts of Cyclospora megacephali n. sp. were subspheroidal, 18.5 X 15.7 (14-21 X 12-18) microns; they had sporocysts pointed at one end with Stieda bodies nearly as wide as the sporocysts themselves, and were 15.0 X 7.2 (11-17 X 6-9) microns; C. megacephali was found in four (31%) hosts. Sporulated oocysts of Eimeria scalopi n. sp. were spheroidal to subspheroidal, 13.6 X 12.6 (11-17 X 11-15) microns with sporocysts lemon-shaped, 8.7 X 5.5 (7-10 X 4-7) microns; it was found in six (46%) hosts. Sporulated oocysts of Eimeria aquatici n. sp. were asymmetrically ellipsoidal, 17.0 X 10.6 (14-20 X 9-14) microns with sporocysts elongately ovoidal, 9.0 X 5.2 (8-11 X 4-6) microns; it was found in two (15%) hosts. Sporulated oocysts of Eimeria motleiensis n. sp. were subspheroidal, 17.0 X 15.3 (15-20 X 13-18) microns with sporocysts ovoidal, 10.7 X 6.8 (10-13 X 6-8) microns; it was found in seven (54%) hosts. Sporulated oocysts of Isospora motleiensis n. sp. were spheroidal to subspheroidal, 13.6 X 12.0 (10-17 X 8-15) microns with sporocysts broadly ovoidal, 9.5 X 6.7 (7-11 X 4-8) microns; it was found in nine (69%) hosts. Sporulated oocysts of Isospora aquatici n. sp. were subspheroidal, 20.9 X 18.4 (15-24 X 13-21) microns with sporocysts ellipsoidal, 11.8 X 9.0 (9-14 X 7-11) microns; it was found in two (15%) hosts.

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PubMed1988

Goussia girellae n. sp. (Apicomplexa: Eimeriorina) in the opaleye, Girella nigricans.

Goussia girellae n. sp. is described from the opaleye fish, Girella nigricans. Merogonic stages were observed in the apices of intestinal epithelial cells, in the lamina propria, and in extra-intestinal sites including liver, gills, and spleen. Gamonts were observed in the intestinal epithelial cells. Only unsporulated oocysts were detected in the intestine, and sporulation occurred when feces containing oocysts were incubated for 48 h in seawater at 21 degrees C. Oocysts are elongated (24.8 x 14.7 micron) with a wall about 200 nm thick and have no residuum, micropyle, or polar granule. Sporocysts are ellipsoid (8.5 x 4.5 micron), have a thin two-layered wall approximately 30 nm thick, and consist of two valves joined by a suture. Although moribund opaleye were also infected with Gyrodactylus sp., Cryptobia sp., Cardicola sp., and epitheliocystis organisms (chlamydia), all fish were heavily infected with G. girellae and morbidity was thus attributed to the coccidium.

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PubMedDate not supplied

Ultrastructure of in vivo-produced caryocysts containing the coccidian Caryospora bigenetica (Apicomplexa: Eimeriidae).

A cotton rat was inoculated orally with oocysts of Caryospora bigenetica from the feces of a rattlesnake. Sixteen days later the rat was euthanized, and portions of the scrotum, foot pad and muzzle were processed for histological sections and transmission electron microscopy. Sporozoites within caryocysts had typical coccidian features such as an anterior and posterior refractile body, centrally located nucleus, micronemes, rhoptries, a conoid, a micropore near the anterior refractile body, a posterior pore, amylopectin granules, lipid bodies, a Golgi-like body, a mitochondrion and subpellicular microtubules. The infected host cell was spherical and surrounded by a fibrous wall-like covering, 0.35-1.00 microns thick. This outer covering, when viewed in stained histological sections, was periodic acid-Schiff (PAS)-positive.

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PubMedDate not supplied

Pathogenesis of Sarcocystis falcatula (Apicomplexa: Sarcocystidae) in the budgerigar (Melopsittacus undulatus). IV. Ultrastructure of developing, mature and degenerating sarcocysts.

Sarcocysts in cardiac and skeletal muscles of budgerigars (Melopsittacus undulatus) were examined transmission electron microscopically 5 to 168 days after experimental infection with Sarcocystis falcatula. The ultrastructure of the primary cyst wall, amorphous substance, metrocytes and bradyzoites in developing, degenerating and mature sarcocysts is described and compared with precystic merozoites studied previously. Sufficient morphologic differences between precystic merozoites, metrocytes and bradyzoites (cystozoites) were found which seem to justify their semantic differentiation. Significant differences in immature and mature primary cyst wall morphology were encountered. If primary cyst wall morphology is to be used in determination and differentiation of species of Sarcocystis, then caution must be used to employ only mature sarcocysts.

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PubMedDate not supplied

Coccidia of Brazilian mammals: Eimeria corticulata n. sp. (Apicomplexa: Eimeriidae) from the anteater Tamandua tetradactyla (Xenarthra: Myrmecophagidae) and Eimeria zygodontomyis n. sp. from the cane mouse Zygodontomys lasiurus (Rodentia: Cricetidae).

Feces from a specimen of Tamandua tetradactyla (Linn.) from Portel, Pará State, north Brazil, contained two different coccidial oocysts; one identified as Eimeria tamanduae Lainson 1968, and the other as a new species, described here as Eimeria corticulata n. sp. Oocysts of E. corticulata are ellipsoidal, 37.4 x 30.4 (31.2-43.7 x 23.7-35.0) microns, shape index (length/width) 1.2 (1.0-1.5). Oocyst wall 2.5-3.7 microns thick and composed of two layers; an outer thick, brown-yellow one with radial striations, and a thin inner smooth one: no visible micropyle. Oocyst residuum a large globule of about 10.7 x 10.3 microns, usually accompanied by a number of smaller attached globules. Sporocysts ellipsoidal, 21.0 x 11.0 (20.0-22.5 x 10.0-12.5) microns, with a conspicuous Stieda body; shape index 1.9 (1.6-2.2). Sporocyst residuum a small number of scattered granules: sporozoites 18.7 x 5.0 microns, with a large posterior refractile body. Eimeria zygodontomyis n. sp. is described in feces from Zygodontomys lasiurus (Lund) from the Serra dos Carajás, Pará. Oocysts ellipsoidal to cylindrical, 16.5 x 12.0 (13.7-18.7 x 11.2-12.3) microns, shape index 1.4 (1.2-1.5). Wall colorless, smooth, single-layered and about 0.6 micron thick: no micropyle. No oocyst residuum, but a polar granule of about 1.8 x 1.0 microns is sometimes present. Sporocysts ellipsoidal, 8.4 x 5.5 (7.5-8.7 x 5.0-6.2) microns, shape index 1.5 (1.4-1.7), with a thin colorless wall and a delicate Stieda body. Sporozoites enclose a compact residuum of about 2.5 x 3.7 microns.

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PubMedDate not supplied

Identification of sporozoite surface proteins and antigens of Eimeria nieschulzi (Apicomplexa).

Sodium dodecyl sulfate polyacrylamide gel electrophoresis, immunoblotting, lectin binding, and 125I surface labeling of sporozoites were used to probe sporozoites of the rat coccidian, Eimeria nieschulzi. Analysis of silver stained gels revealed greater than 50 bands. Surface iodination revealed about 14 well labeled, and about 10 weakly labeled but potential, surface proteins. The most heavily labeled surface proteins had molecular masses of 60, 53-54, 45, 28, 23-24, 17, 15, 14, 13, and 12 kD. Following electrophoresis and Western blotting, 2 of the 12 125I labeled lectin probes bound to two bands on the blots, which collectively indicated that two bands were glycosylated. Concanavalin A (ConA) specifically recognized a band at 53 kD, which may represent a surface glycoprotein, and a lectin derived from Osage orange (MPA) bound to a single band at 82-88 kD, that may also be a surface molecule. Immunoblotting using sera collected from rats inoculated orally with oocysts, as well as sera from mice hyperimmunized with sporozoites, revealed that many surface molecules appear to be immunogenic.

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PubMedDate not supplied

Coccidia of Brazilian mammals: Eimeria marajoensis N. Sp. (Apicomplexa: Eimeriidae) from the Anteater, Tamandua tetradactyla (Xenarthra: Myrmecophagidae).

Feces from a juvenile specimen of the anteater Tamandua tetradactyla from Ponta de Pedras, Marajó, Pará, northern Brazil, contained three different coccidial oocysts: Eimeria tamanduae Lainson, 1968; E. corticulata Lainson & Shaw, 1990; and a third species previously unrecorded and described here as Eimeria marajoensis n. sp. Oocysts of the latter parasite are spherical to subspherical, 13.9 +/- 1.5 x 13.4 +/- 1.4 (11.1-16.5 x 11.1-16.5) microns, shape index (length/width) 1.0 (1.0-1.2). The oocyst wall is a single, colorless layer about 0.6-1.0 microns thick with no striations or micropyle. There is no oocyst residuum, but a single, round, oval or irregularly shaped polar granule of about 0.75-2.5 microns is consistently present. The sporocysts are broadly ellipsoidal, 7.1 +/- 0.7 +/- 5.3 +/- 0.6 (6.0-8.8 x 4.0-5.7) microns, shape index 1.3 (1.2-1.5), with a delicate wall bearing minute stieda body. No sub-stieda body was visible. The sporocyst residuum consists of some 10-20 rounded granules, lying between the two slightly curved sporozoites which measure approximately 6.5 x 2.0 microns. Sporocyst refractile bodies were not discernable.

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PubMedDate not supplied

Gametogenesis and sporogony of Hepatozoon mocassini (Apicomplexa: Adeleina: Hepatozoidae) in an experimental mosquito host, Aedes aegypti.

The sexual life cycle of the hemogregarine Hepatozoon mocassini was studied in Aedes aegypti, an experimental mosquito host, using transmission electron microscopy. Gamonts were observed leaving the host snake erythrocyte as early as 30 min after mosquitoes ingested infected blood, and some gamonts had penetrated the gut epithelial cells by this time. Six hours post-feeding, gamonts were identified within cells of the abdominal fat body. Twenty-four hours post-feeding, gamonts were often entrapped within the peritrophic membrane, but were no longer observed within the gut wall. Parasites pairing up in syzygy and undergoing sexual differentiation were observed within fat cells at this time, and by 48 hours post-feeding, well-developed macro- and microgametocytes as well as microgametes were discernible. Developing zygotes observed 3 days post-feeding were enclosed within a parasitophorous vacuole. By day 6, multinucleate oocysts with crystalloid bodies in the cytoplasm were seen. Sporozoites developing within sporocysts appeared by day 12. Seventeen days post-feeding, mature oocysts with sporocysts containing approximately 16 sporozoites were observed upon dissection of mosquitoes. Large crystalloid bodies no longer bound by rough endoplasmic reticulum were located anterior and posterior to the sporozoite nucleus. Free sporozoites were not observed.

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PubMedDate not supplied

A comparative study of lipid compositions of Cryptosporidium parvum (Apicomplexa) and Madin-Darby bovine kidney cells.

Membrane lipid compositions of Cryptosporidium parvum and Madin-Darby bovine kidney cells, an epithelial-like cell line commonly used to study coccidia in vitro, were analyzed using both thin-layer chromatography and gas-liquid chromatography. Phosphatidylcholine was the predominant lipid in both C. parvum and Madin-Darby bovine kidney cells, comprising 65% and 41% of the total phospholipids, respectively. Phospholipids of C. parvum contained twice the level of 16:0 and twenty-fold more 18:2 than the Madin-Darby bovine kidney cell line. We suggest that the parasite may be capable of sequestering specific complex membrane lipids at concentrations greater than those in the host cells. This study constitutes the first report of the lipid composition of C. parvum.

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PubMedDate not supplied

Isospora thibetana N. sp. (Apicomplexa, Eimeriidae), a parasite of the Tibetan siskin (Serinus thibetanus = Carduelis thibetanus) (Passeriformes, Fringillidae).

Tibetan siskins are birds native to the Himalayan region often imported into Italy for commercial purposes. Fecal examination of 45 imported subjects with clinical signs of diarrhoea revealed the presence of a large number of coccidian oocysts. After sporulation, accomplished by mixing feces with 2.5% (w/v) aqueous K2Cr2O7 at room temperature (22 degrees C +/- 1 degree C), exogenous stages of an Isospora species were revealed. The oocysts of this Isospora are spherical, have a bilayered colorless wall, and average 23.24 microm x 23.05 microm; oocyst residuum and micropyle are absent, while an oval polar granule is rarely present. The elliptical sporocysts average 18.44 microm x 10.97 microm and the Stieda body protrudes slightly from the end of the sporocyst. A spherical sporocyst residuum, is present though it sometimes consists of scattered granules. The spindle-shaped sporozoites average 11.53 microm x 2.86 microm, and have two refractile bodies. The taxonomic position of the tibetan siskin is controversial. Some authors include this species in the genus Serinus, while others include it in the genus Carduelis. The coccidian species isolated from these tibetan siskins was, for this reason, compared with the Isospora species previously described both in the genus Carduelis and in the genus Serinus. As a result of this comparison a new species, Isospora thibetana, was named. In the intestine of dead subjects, oocysts were found only in the ileum where the mucosa was greatly thickened and presented a heavy leucocytic infiltration consisting mainly of lympho-monocytic cells. A similar infiltration was observed in liver and lungs as well.

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PubMedDate not supplied

Ultrastructural features of cystic and merogonic stages of Hepatozoon sipedon (Apicomplexa: Adeleorina) in northern leopard frogs (Rana pipiens) and northern water snakes (Nerodia sipedon) from Ontario, Canada.

The cystic and merogonic stages of the haemogregarine Hepatozoon sipedon, infecting Northern water snakes (Nerodia sipedon sipedon) and Northern leopard frogs (Rana pipiens), respectively, in Ontario, Canada, were investigated by transmission electron microscopy. Cysts, which were observed in the liver of Northern leopard frogs (Rana pipiens) after these anurans ingested mosquitoes (Culex pipiens) containing oocysts of the parasite, harboured two cystozoites, each of which contained a large crystalloid inclusion anterior to the nucleus. Two types of meronts were observed in snakes that were fed the liver of infected frogs. Macromeronts, which matured in endothelial cells of the liver approximately 16 d after snakes ingested infected frogs, contained about 50 large macromerozoites. Macromerozoites emerged from macromeronts, entered the bloodstream of the snake, and reinfected endothelial cells. Micromeronts, which matured about 34 d post-inoculation, contained about 150 micromerozoites that infected erythrocytes and transformed into gamonts. The ultrastructural features of micromeronts and macromeronts differed only slightly: immature macromeronts and macromerozoites contained numerous amylopectin and lipid inclusions, whereas immature micromeronts and micromerozoites did not contain amylopectin inclusions and featured fewer, smaller lipid inclusions. A comparison of cystic stages among Hepatozoon species in different groups of vertebrates is presented with respect to their structure and evolutionary significance.

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PubMedDate not supplied

Cryptosporidium andersoni n. sp. (Apicomplexa: Cryptosporiidae) from cattle, Bos taurus.

A new species of Cryptosporidium is described from the feces of domestic cattle, Bos taurus. Oocysts are structurally similar to those of Cryptosporidium muris described from mice but are larger than those of Cryptosporidium parvum. Oocysts of the new species are ellipsoidal, lack sporocysts, and measure 7.4 x 5.5 microm (range, 6.0-8.1 by 5.0-6.5 microm). The length to width ratio is 1.35 (range, 1.07-1.50). The colorless oocyst wall is < 1 microm thick, lacks a micropyle, and possesses a longitudinal suture at one pole. A polar granule is absent, whereas an oocyst residuum is present. Oocysts were passed fully sporulated and are not infectious to outbred, inbred immunocompetent or immunodeficient mice, chickens or goats. Recent molecular analyses of the rDNA 18S and ITS1 regions and heat-shock protein 70 (HSP-70) genes demonstrate this species to be distinct from C. muris infecting rodents. Based on transmission studies and molecular data, we consider the large form of Cryptosporidium infecting the abomasum of cattle to be a new species and have proposed the name Cryptosporidium andersoni n. sp. for this parasite.

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PubMedDate not supplied

Cryptosporidium hominis n. sp. (Apicomplexa: Cryptosporidiidae) from Homo sapiens.

The structure and infectivity of the oocysts of a new species of Cryptosporidium from the feces of humans are described. Oocysts are structurally indistinguishable from those of Cryptosporidium parvum. Oocysts of the new species are passed fully sporulated, lack sporocysts. and measure 4.4-5.4 microm (mean = 4.86) x 4.4-5.9 microm (mean = 5.2 microm) with a length to width ratio 1.0-1.09 (mean 1.07) (n = 100). Oocysts were not infectious for ARC Swiss mice, nude mice. Wistar rat pups, puppies, kittens or calves, but were infectious to neonatal gnotobiotic pigs. Pathogenicity studies in the gnotobiotic pig model revealed significant differences in parasite-associated lesion distribution (P = 0.005 to P = 0.02) and intensity of infection (P = 0.04) between C. parvum and this newly described species from humans. In vitro cultivation studies have also revealed growth differences between the two species. Multi-locus analysis of numerous unlinked loci, including a preliminary sequence scan of the entire genome demonstrated this species to be distinct from C. parvum and also demonstrated a lack of recombination, providing further support for its species status. Based on biological and molecular data, this Cryptosporidium infecting the intestine of humans is proposed to be a new species Cryptosporidium hominis n. sp.

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PubMedDate not supplied

Redescription of the sarcocysts of Sarcocystis rileyi (Apicomplexa: Sarcocystidae).

The intermediate hosts for Sarcocystis rileyi (Stiles 1893) Minchin 1913 are ducks (Anas spp.), and the striped skunk (Mephitis mephitis) is its definitive host. The structure of sarcocysts from an experimentally infected shoveler duck (Anas cylpeata) fed sporocysts from an experimentally-infected M. mephitis was studied and compared with type specimens from a naturally infected duck. The experimentally infected duck was killed 154 d after feeding sporocysts. By light microscopy the sarcocyst wall was 3-5 microm thick with indistinct villar protrusions. Ultrastructurally, the sarcocyst wall was a type-23 cyst wall with anastomosing villar protrusions that were up to 7.5 microm long. The villar projections contained filamentous structures. The bradyzoites were 12-14 microm long. Structurally, the sarcocyst from the naturally infected and experimentally infected ducks appeared similar.

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PubMed1994

A simple and reliable method of producing in vitro infections of Cryptosporidium parvum (Apicomplexa).

A variety of techniques have been used to infect cell monolayers in culture with the protozoan, Cryptosporidium parvum. However, most of these methods rely on the use of trypsin and/or bile salts to excyst sporozoites in vitro, followed by washing sporozoites free of excystation solution prior to their addition to subconfluent monolayers. This method not only increases the amount of time required to establish infections in vitro, but also results in prolonged exposure of free sporozoites to environmental conditions. Here we report a simple, fast, and efficient method of obtaining consistent infections of C. parvum in cell monolayers. This technique relies on the ability of the parasite to excyst at 37 degrees C but not at room temperature following pretreatment with sodium hypochlorite. By adding surface-sterilized oocysts directly to monolayers, sporozoites have access to host cells immediately upon excystation.

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PubMed1994

Comparative development of Cryptosporidium parvum (Apicomplexa) in 11 continuous host cell lines.

Using standardized media, incubation, and parasite inoculating procedures, we compared development of Cryptosporidium parvum between Madin-Darby bovine kidney (MDBK) cells and 10 additional host cell lines available through the American Type Culture Collection. Parasite development was assessed by counting parasite numbers atop monolayers in 25 random oil fields 68 h post-infection using Nomarski interference-contrast optics. Results revealed that the human ileocecal adenocarcinoma (HCT-8) cell line supported nearly twice the number of parasite developmental stages than MDBK cells or any of the other host cell types.

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PubMed1989

Major histocompatibility complex control of immunity elicited by genetically engineered Eimeria tenella (Apicomplexa) antigen in chickens.

The immunogenicity of a recombinant Eimeria tenella coccidial antigen was studied in 6(1).B congenic chickens derived from B2B2 and B5B5 parents segregating for haplotypes B2 and B5. Five-week-old chickens were immunized with 2.4 micrograms of recombinant protein (designated 5401) in Freund complete adjuvant and challenged with 75,000 oocysts at 28 days postimmunization (DPI) to determine the degree of elicited protective immunity. Serum samples were collected weekly for 5 weeks postimmunization for analysis by enzyme-linked immunosorbent assay, immunofluorescence assay, and Western blotting. Lesion scores following oocyst challenge were significantly reduced in B5B5 chickens compared with those in B2B2 chickens. Immunization induced a sporozoite-specific immunoglobulin G (IgG) titer in serum detected by the enzyme-linked immunosorbent assay that peaked at 28 DPI, the day of challenge, in B5B5 chickens and at 42 DPI in B2B2 chickens. After challenge, this titer declined for each genotype. Anti-sporozoite IgG detected by the immunofluorescence assay attained a peak titer at 21 DPI in B2B2 chickens and 28 DPI in B5B5 chickens. Serum from immunized B5B5 chickens reacted strongly in Western blots with several high-molecular-weight (greater than 100,000), soluble proteins prepared from sporozoites. Serum from B2B2 chickens reacted with similar proteins as well as with a 51- to 53-kilodalton protein that was not labeled by serum from B5B5 chickens. These results demonstrate further the role of host genetics on anticoccidial immunity and suggest that a peak anti-sporozoite IgG titer in B5B5 chickens on the day of challenge may signal a state of immunocompetence to that challenge.

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