Life Cycle and Host Specificity of Eimeria callospermophili Henry, 1932 From the Uinta Ground Squirrel Spermophilus armatus*

SYNOPSIS. Eimeria callospermophili was found in 6 species of ground squirrels and the white-tailed prairie dog. The hosts included Spermophilus armatus from Utah and Montana, S. richardsoni from Montana and Wyoming, S. beecheyi from California, S. lateralis and S. variegatus from Utah, and S. tridecemlineatus and Cynomys leucurus from Wyoming. Infections were generally transmissible from each species of ground squirrel to S. armatus and S. richardsoni. Oocysts from C. leucurus caused infections in S. armatus and S. richardsoni. No infections were found after inoculation of E. callospermophili oocysts into least chipmunks (Eutamius minimus), Mongolian gerbils (Meriones unguiculatus), or laboratory rats; however, excystation occurred in these animals. Resistance to infection did not develop in S. armatus, S. richardsoni, or S. variegatus, but did occur after 5 or more infections in S. lateralis. Eimeria callospermophili had little or no effect on the host in S. armatus, S. lateralis, or S. variegatus, but caused bloody diarrhea in severely infected individuals of S. richardsoni. The oocysts had an oocyst residuum consisting of several distinct bodies, which later coalesced to form a large homogeneous body. Each sporozoite had an unusually large refractile body. In experimentally infected specimens of S. armatus the prepatent period and patent period lasted for 5 and 9 days, respectively. Mature 1st-generation schizonts, first seen 2 days after inoculation, had 8–12 merozoites. Mature 2nd-generation schizonts, first seen 3 days after inoculation, had an average of 18 merozoites which were smaller than those of the 1st generation. Mature gametes were 1st seen 4 days after inoculation. Mature microgametocytes were only slightly larger than mature macrogametes.     

Observations on the Life Cycle of Eimeria bilamellata Henry, 1932 in the Uinta Ground Squirrel Spermophilus armatus*

SYNOPSIS. Oocysts of Eimeria bilamellata were found in Spermophilus armatus, Utah and Wyoming; S. beecheyi, California; and S variegatus, Utah. Oocysts were not found in S. lateralis, S. richardsoni, S. columbianus, S. tridecemlineatus, or the white-tailed prairie dog, Cynomys leucurus. Experimental infections were established in S. armatus, S. variegatus, and S. lateralis, but not in S. richardsoni, least chipmunks Eutamius minimus, laboratory rats Rattus norvegicus, or Mongolian gerbils Meriones unguiculatus. After one experimental infection S. armatus and S. variegatus were immune to further infections. Spermophilus lateralis could be infected 3 or 4 times before the animals were immune. However, individuals of S. armatus in a natural population had more than one infection with E. bilamellata; probably infections must be of a certain level before immunity develops. When S. armatus were inoculated with about 100,000 oocysts, the animals usually died on the 7th day after incoculation. Oocysts were 33-37 by 25-31 μ (mean 34.5 by 28.2 μ). The oocyst wall was brown and composed of 2 layers. A distinct micropyle was present. Sporocysts were 18-23 by 9-12 μ (mean 19.9 by 10.3 μ). In experimental infections, the prepatent period was 10 days and the patent period 5–21 (mean 9) days. Schizonts were 1st seen 7 days after inoculation. They were located above the host cell nuclei of epithelial cells at the tips of the villi of the jejunum and ileum. One or more earlier generations of schizonts were thought to occur, but these were not observed. Gametogony took place in epithelial cells of the jejunum and ileum. Shortly after the merozoites entered the cells, the cells became enlarged and were displaced into the lamina propria. The microgametocytes were considerably larger than the macrogametes and contained a central residual body. Macrogametes had a peripheral eosinophilic layer as well as cytoplasmic granules; both apparently participated in formation of the oocyst wall.     

The Life Cycle of Eimeria vermiformis Ernst,Chobotar and Hammond, 1971 in the Mouse Mus musculus1

SYNOPSIS. The life cycle of Eimeria vermiformis from the mouse Mus musculus is described from experimental infections. The prepatent period was 7 days, and the patent period 7–22+ days. Endogenous stages were in the lower 2/3 of the small intestine. Two generations of schizonts were found. Mature 1st generation schizonts, seen 4 days after inoculation, were 16–25 × 9–16 μ and had long vermiform merozoites. Mature 2nd generation schizonts were first seen 5 days after inoculation. They were 8–18 × 7–14 μ (mean 11.2 × 13.1 μ). Mature microgametocytes, 17–32 × 12–25 μ, were present 6 days after inoculation. Macrogametes with plastic granules were found at the same time. The life cycle of E. vermiformis is compared with those of other species of Eimeria from Mus.     

Development of Eimeria callospermophili and E. bilamellata from the Uinta Ground Squirrel Spermophilus armatus in Cultured Cells*

SYNOPSIS. Cell lines or established cell lines of bovine, ovine or human origin and primary cells from whole embryos of groundsquirrels were used in a study of the in vitro development of Eimeria callospermophili and E. bilamellata from the Uinta ground squirrel, Spermophilus armatus. Monolayers in Leighton tube cultures were inoculated with sporozoites of either of these 2 species and examined with phase-contrast microscopy at various intervals. After such examination, coverslips were fixed in Schaudinn's or Zenker's fluid and variously stained. E. callospermophi sporozoites penetrated cells and underwent development to mature 1st generation schizonts in most cell types. At different times after inoculation, both species formed sporozoite-shaped schizonts, which later became spheroidal. Intracellular movements of sporo zoite-shaped schizonts of E. callospermophili were observed and such schizonts penetrated cells when freed by mechanical disintegration of the host cells. Merozoites were formed at the periphery of the schizont in both species. Mature 1st generation schizonts of E. callospermophili, with 6–14 merozoites, were first seen 15 hr after inoculation; the corresponding values for E. bilamellata were 12–27 merozoites and 4 days. Merozoites of both had anterior and posterior refractile bodies. Exposure to a trypsin-bile solution stimulated motility in merozoites of E. callospermophili. Second generation trophozoites and immature schizonts of E. callospermophili were seen in cultures of primary cells of whole ground-squirrel embryos 20–24 hr and 44–48 hr, respectively, after inoculation of sporozoites.     

Life Cycle of Isospora canis Nemeséri, 1959 in the Dog*

The life cycle of I. canis Nemeséri, 1959 was studied in experimentally infected dogs. Freshly sporulated oocysts were ovoid and 34–40 × 28–32 μm. The endogenous stages were found directly beneath the epithelium of the distal portion of the small intestinal villi. Most of the endogenous stages were in the lower 1/3 of the small intestine, but occasionally they were found in other portions of the small intestine. Three asexual generations were present. First-generation schizonts were 16–38 × 11–23 μm and contained 4–24 merozoites; mature 1st-generation merozoites were 8–11 × 3–5 μm. First-generation schizogony lasted up to 7 days after inoculation. Second-generation schizonts were 12–18 × 8–13 μm and contained up to 12 merozoites which were 11–13 × 3–5 μm. Second-generation schizogony was present on postinoculation days 6 and 7. Third-generation schizonts were formed by nuclear division of 2nd-generation merozoites. Most 2nd-generation merozoites underwent nuclear division without leaving the parasitophorous vacuole of the 2nd-generation schizont. Mature 3rd-generation schizonts were 13–38 × 8–24 μm and contained 6–72 merozoites. Third-generation merozoites were 8–13 × 1–3 μm. Third-generation schizogony was present on days 6–8 after inoculation. Mature macrogametes were 22–29 × 14–23 μm. Mature microgametocytes were 20–38 × 14–26 μm. Gametes were present on postinoculation days 7–10. Oocysts were present in tissue sections on postinoculation days 8–10 and 12. The prepatent period was 9–11 days.     

In vitro Development of First-Generation Schizonts of Eimeria canadensis (Bruce, 1921)*†

SYNOPSIS. In vitro development of Eimeria canadensis from cattle was studied in monolayer cultures of various bovine cell lines grown on coverslips in Leighton tubes. Excysted sporozoites were used for inoculation of the cell cultures. Sporozoites entered the host cells within a few minutes, but apart from a reduction in the number of refractile bodies, changed little in appearance during the first 9 days. Beginning at 91/2 days postinoculation, sporozoites developed into sporozoite-shaped schizonts or, less frequently, transformed into trophozoites. Sporozoite-shaped schizonts with as many as 8 nuclei were observed transforming into spheroid schizonts. At 111/2 days, intermediate schizonts had a characteristic single mass of refractile granules and 60–80 nuclei. Deep invaginations, which resulted in the formation of several blastophores, usually occurred when schizonts had about 100 nuclei. Merozoites were formed as a result of radial outgrowth from the surface of spheroid schizonts as well as of blastophores. Mature merozoites were seen 1st after 13 days.     

Development of Eimeria auburnensis in Cell Cultures

SYNOPSIS. Monolayer established cell line cultures of bovine kidney (Madin-Darby) and human intestine (Intestine 407), as well as embryonic bovine tracheal and embryonic spleen cell line cultures were inoculated with E. auburnensis sporozoites and observed for a maximum of 22 days. Mature 1st generation schizonts developed in the kidney, tracheal and spleen cells. In the intestine cells, trophozoites were seen in 3 of 4 experiments, but schizonts were not found. Sporozoites penetrated cells, beginning within a few minutes after inoculation. Penetration was usually accomplished within 10 seconds, and the body of the sporozoite underwent a slight constriction as it passed thru the host cell membrane. Some sporozoites left cells. Numerous intracellular sporozoites were observed in kidney, tracheal and spleen cultures. Crescent bodies were seen in the parasitophorous vacuole as early as 1 day after inoculation. At this time, the nuclei of most intracellular sporozoites had changed from vesicular to compact. Beginning 4 days after inoculation, enlarged sporozoites and parasites having a sporozoite shape, but with 2-5 nuclei, were frequently seen. These enlarged sporozoites and sporozoite-shaped schizonts evidently transformed into trophozoites and spheroidal schizonts by means of lateral outpocketings. Few trophozoites were seen. More immature schizonts developed in kidney cells than in the other cell types. The numbers of mature schizonts observed in kidney and tracheal cells were similar, but development occurred less consistently in the latter. Few immature and mature schizonts developed in spleen cells. Mature schizonts, first seen 9 days after inoculation, were considerably smaller than those reported from calves. Some motile merozoites were seen; evidently no development beyond these occurred. The nucleus and nucleolus of host cells were enlarged; this enlargement was not as pronounced as in infections in calves. Multiple host cell nuclei were frequently observed. Degenerative changes in the cultured cells and in the parasites usually occurred, beginning 9-17 days after inoculation; these were more pronounced in the spleen cells than in the others.     

The Life Cycle of Isospora felis Wenyon, 1923, a Coccidium of the Cat*

SYNOPSIS. A pure strain of Isospora felis derived from a single oocyst was used to study the endogenous cycle. One and a half to two-month-old laboratory-reared, coccidia-free kittens were used thruout the study. The endogenous stages occurred in the epithelial cells of the distal parts of the villi in the ileum and occasionally duodenum and jejunum. All stages lay above the host cell nucleus. There were 3 asexual generations. The 1st generation schizonts were 11–30 by 10–23 μ when mature and contained 16–17 banana-shaped merozoites 11–15 by 3–5 μ. They became mature in 96 or sometimes in 120 hours. The 1st generation merozoites entered new host cells, rounded up and formed 2nd generation schizonts. These formed within themselves 2–10 or more spindle-shaped bodies resembling 1st generation merozoites in shape and size. These were 2nd generation merozoites. They were uninucleate 120 hours after inoculation, but by 144 hours they became larger, multinucleate and some lost their elongate shape and became ovoid. They were then 3rd generation schizonts. They were 12–16 by 4–5 μ. Each formed up to 6 or more banana-shaped merozoites 6–8 by 1–2 μ. The 3rd generation schizonts and merozoites developed within the same host cell and parasitophorous vacuole as the 2nd generation schizonts and merozoites. Mature schizonts containing only 3rd generation merozoites appeared 144 hours after inoculation, were most abundant 168 hours after inoculation, and might be present as late as 216 hours after inoculation. They were 14–36 by 13–22 μ and contained 36 to more than 70 merozoites. The 3rd generation merozoites entered the sexual cycle. The mature microgametocytes were 24–72 by 18–32 μ and contained a central residuum and a large number of microgametes 5–7 by 0.8 μ with 2 posteriorly-directed flagella. The mature macrogametes were 16–22 by 8–13 μ. Gametogony occurred 144–216 hours after inoculation. The prepatent period was 168–192 hours and the patent period 10–11 days. Peak oocyst production occurred on the 6th day of the patent period.     

Development of Eimeria alabamensis from Cattle in Mammalian Cell Cultures*

SYNOPSIS. Monolayer primary cultures of cells from bovine embryonic intestine (BEInt), kidney (BEK), spleen (BES), and thyroid (BETy) and cell line cultures of embryonic bovine trachea (EBTr) and synovium (BESy) as well as established cell line cultures of bovine kidney (Madin-Darby, MDBK), human intestine (Int 407) and Syrian hamster kidney (BHK) were inoculated with freshly excysted sporozoites of Eimeria alabamensis and observed for 4–5 days. Sporozoites penetrated all cell types; during the 1st 24 hr, intracellular sporozoites, trophozoites and binucleate schizonts were seen in all cell cultures. Mature schizonts were more numerous in BES and MDBK cells than in the others. Large schizonts, 14.2 (11–18.5) by 10.2 μ (8.5–11), with 6–14 short, stubby merozoites (each with 2 refractile bodies) occurred at 2 and 3 days in all cells except BESy, Int 407, and BHK. Small schizonts, 9.7 (5.5–13) by 6 μ (5–8.5), with 6–10 long, slender merozoites (each with 2 refractile bodies) were found 3 days after inoculation in all cell types. At 4 days, some intracytoplasmic merozoites and a few intranuclear 2nd generation trophozoites were found. After 4 days post-inoculation, intracellular parasites were rarely seen and these were apparently degenerate. Development within the host cell nucleus, the normal site of development in the host animal, was observed infrequently in cell cultures. Intranuclear sporozoites, found no earlier than 2 days after inoculation, developed similarly to those in the cytoplasm, and small intranuclear schizonts with 6–10 merozoites (each with 2 refractile bodies) occurred after 3 days in culture.     

The life cycle of Eimeria falciformis var. pragensis (Sporozoa: Coccidia) in the mouse, Mus musculus

The life cycle of Eimeria falciformis var. pragensis, established from a single oocyst, is described in experimentally infected mice (Mus musculus). The coccidium had a prepatent period of 7 days and a patent period of 10--16 days. Oocysts were spherical to ellipsoidal in shape and measured 21.2 x 18.3 micron. Sporulation time was 3 to 3.5 days. Sporocysts measured 12.2 x 7.2 micron and contained a circular to avoid granular sporocyst residuum measuring 5.5 X 5.0 micron. One, 2 or 3 circular to rectangular polar granules were observed within each sporulated oocyst. The endogenous stages developed primarily in the cecum and colon and only occasionally in the lower ileum. Four generations of schizonts were found. Mature 1st-generation schizonts, first observed 48 hr postinfection (PI), measured 17.8 x 12.3 micron and had 12 merozoites that measured 13.3 x 2.0 micron. Mature 2nd-generation schizonts appeared 78 hr PI. They measured 10.2 x 9.3 micron and had 8 merozoites measuring 5.0 x 1.6 micron. Mature 3rd-generation schizonts appeared first at 114 hr PI and measured 17.5 x 10.2 micron and had 10 merozoites that measured 12.4 x 1.8 micron. Mature 4th-generation schizonts appeared first at 144 hr PI. They measured 18.2 x 15.3 micron and had 18 merozoites. The merozoites of the 4th-generation schizont were 4.5 x 1.2 micron. Mature macrogamonts and microgamonts developed simultaneously appearing at 156 hr PI. Macrogamonts measured 16 x 14.5 micron and microgamonts were 18.2 x 15.3 micron. In experimentally infected rats (Rattus norvegicus), development of E. falciformis var. pragensis progressed only as far as mature 1st-generation schizonts.     

Development of Eimeria meleagrimitis Tyzzer from Sporozoites and Merozoites in Turkey Kidney Cell Cultures*

SYNOPSIS. Sporozoites and 1st-, 2nd-, and 3rd-generation merozoites of Eimeria meleagrimitis were inoculated into primary cultures of turkey kidney cells. In vitro-excysted sporozoites developed into mature macrogamonts in 8 days; in vivo-excysted sporozoites developed into 2nd- or 3rd-generation schizonts within 5 to 7 days. First-generation merozoites obtained from infected turkeys produced mature 2nd-generation schizonts within 24 h. Second-generation merozoites from turkeys produced mature macrogamonts and oocysts within 72 h, whereas 3rd-generation merozoites produced these stages within 48 h. The oocysts that developed from 3rd-generation merozoites sporulated at 25 C and were infective for turkeys. The timing of the early stages and the intervals between schizogonic generations in cultures were comparable with those in turkeys. Morphologic parameters, however, indicated that some differences existed between in vitro and in vivo development. Second- and 3rd-generation schizonts and gamonts that developed after inoculation of cultures with merozoites were similar to stages in turkeys. Oocysts, however, were significantly smaller (P < 0.05) in cultures. All stages that developed after inoculation of cultures with sporozoites were smaller (P < 0.05) than their in vivo counter parts.     

Ultrastructural Study of Schizogony in Eimeria callospermophili*

SYNOPSIS The development of 1st generation schizonts of Eimeria callospermophili was studied with cell cultures and with experimentally infected host animals, Spermophilus armatus. Sporozoite-shaped schizonts each had 5-10 nuclei and all of the organelles of the sporozoite; each nucleus had a nucleolus and an associated Golgi apparatus. In stages immediately preceding merozoite formation, an intranuclear spindle apparatus with conical polar areas were observed near the outer margin of each nucleus. Two centrioles, each having 9 single peripheral tubules and one central tubule, were observed near each pole in some specimens. Merozoite formation began internally, with anlagen of 2 merozoites developing near each nucleus. The inner membrane of the merozoites first appeared as 2 dense thickenings adjacent to the polar cones and centrioles; subpellicular microtubules appeared simultaneously. Two anterior annuli and the conoid formed between the 2 thickenings. Vesicles, possibly of Golgi origin, were located next to the forming inner membrane. As the forming merozoites underwent elongation, a rhoptries anlage, a Golgi apparatus, refractile bodies, and mitochondria were incorporated into each. Sporozoite-shaped schizonts with merozoite anlagen transformed into spheroid or ovoid schizonts; at this time the conoid, rhoptries, micronemes, and the inner membrane of the pellicle gradually disappeared; several small refractile bodies were formed from the larger one. When development was about 1/3 complete, the immature merozoites began to grow outward from the surface of the schizont. In this phase of development, the single surface membrane of the schizont became the outer membrane of the merozoite's pellicle, and additional organelles, including the nucleus, were incorporated. Finally, the merozoites became pinched off, leaving a residual body. Development in cell cultures and host tissues was similar. This type of schizogony, previously undescribed in Eimeria, is compared with corresponding stages of development in other species of Eimeria and Sporozoa.     

Observations on the Endogenous Stages of Eimeria confusa Joseph, 1969 from the Grey Squirrel Sciurus carolinensis*

SYNOPSIS. Stages in the endogenous cycle of Eimeria confusa from the grey squirrel, Sciurus carolinensis, are described from mixed infections with another species, Eimeria lancasterensis. All corresponding stages were markedly different in the 2 species. In E. confusa infections, the parasites were located below the host cell nuclei of the epithelial cells of the villi of the jejunum and ileum. Mature schizonts were ellipsoidal, averaged 20.9 × 18.6 μm and had 18–30 merozoites. The mature microgamonts measured 34.3 × 24.7 μm and had hundreds of microgametes. Mature macrogametes were ovoid, averaged 31.3 × 25.6 μm, and contained 2 kinds of plastic granules.     

Life Cycle of Eimeria ferrisi Levine & Ivens, 1965 in the Mouse, Mus musculus

SYNOPSIS. The life cycle of Eimeria ferrisi is described from experimentally infected Mus musculus. The prepatent period was 3 days and the patent period was 3–4 days. The endogenous stages were found only in the cecum and colon. Three generations of schizonts were found. Mature 1st-generation schizonts first seen 24 hr postinoculation (PI) measured 10.9 (7–14) × 10.2 (6–13) μm and had 9.6 (7–14) merozoites. Some 2nd-generation schizonts had uninucleate merozoites and others had multinucleate merozoites. The former were first seen in small numbers 36 hr PI and were most abundant 48 hr PI. They measured 9.6 (5–13) × 7.9 (6–12) μm and had 18 (6–25) merozoites. Schizonts with multinucleate merozoites were seen 72 hr PI. Mature 3rd-generation schizonts were seen 72 hr PI. They measured 14.0 (12–18) × 11-0 (9–13) μm and had 12.5 (5–16) merozoites. Macrogamonts were first seen in 72 hr sections. Each young macrogamont had a large nucleus with a prominent nucleolus. Only one type of cytoplasmic granule appeared to be involved in the formation of the oocyst wall. Mature macrogamonts were 11.0 (5–14) × 10.0 (6–13) μm. Crescent-shaped bodies were observed in the parasitophorous vacuole of trophozoites and young macrogamonts. Early microgamonts were first recognized at 96 hr by the presence of darkly stained and irregularly shaped nuclei. Usually, mature microgametes were arranged in long, narrow whorls at the periphery of the microgamont or in whorls at the surface of 2–5 compartments.     

Development of Eimeria ninakohlyakimovae from Sheep In Cell Cultures*

SYNOPSIS. Monolayer cell line cultures of ovine trachea, thyroid, thymus, and kidney cells, as well as an established cell line (Madin-Darby) of bovine kidney cells, were inoculated with sporozoites of Eimeria ninakohlyakimovae and observed for a maximum of 24 days. Sporozoites were seen penetrating cells within 5 minutes after inoculation, as well as 2 and 3 days after inoculation, and leaving cells 3 days after inoculation. Transformation from sporozoites to trophozoites occurred by a widening or by a lateral outpocketing of the sporozoite body. Trophozoites and schizonts were first seen 3 days after inoculation in all ovine cell types. Large numbers of immature schizonts were observed, but only an estimated 0.4–4.3% of these became mature in the different kinds of cells. Usually, mature schizonts were first seen 10–11 days after inoculation in the ovine cells, but they sometimes occurred as early as 8 days. More mature schizonts were seen in the ovine kidney and trachea cells than in the others; the smallest number occurred in the bovine cells. The nucleoli of cells harboring large schizonts in each type of culture were enlarged and the chromatin clumps normally seen in the nuclei of non-infected cells were not visible. The cytoplasm of some infected cells was vacuolated. The formation of merozoites occurred by a budding process from blastophores, from the surface of schizonts, and/or from infoldings and invaginations of this surface. Merozoites were observed leaving host cells, but were not seen penetrating new cells. Intracellular first-generation merozoites were observed 13 and 15 days after inoculation in lamb trachea and kidney cells, respectively. No evidence of further development of such merozoites was found.     

Schizogony and Gametogony of Leucocytozoon simondi and Associated Reactions in the Avian Host*

SYNOPSIS. Stages of development of Leucocytozoon simondi in White Pekin ducklings and their reactions to the parasite were studied on successive days after infecting them artificially with sporozoites from Simulium rugglesi. The minimum prepatent period was 5 days. The first asexual cycle occurred exclusively in the parenchymal cells of the liver. Progeny of these hepatic schizonts followed one of 3 courses: (a) invaded parenchymal liver cells to give rise to another hepatic cycle, (b) penetrated blood cells to form round gametocytes, and (c) were phagocytized by macrophages and grew into megaloschizonts thruout the body. The appearance of elongating gametocytes coincided with the period of maturation and release of merozoites from the megaloschizonts. Experimental evidence supports the hypothesis that the round gametocytes arise from the hepatic schizonts and the elongate forms from the megaloschizonts. Mature megaloschizonts released millions of merozoites, but a high 2nd peak in parasitemia did not develop because of retention of developing gametocytes in the deep circulation, particularly the liver and spleen, and a pronounced host reaction.     

Development of First-Generation Schizonts of Eimeria bovis in Cultured Bovine Cells

SYNOPSIS. Monolayer primary and secondary cultures of embryonic bovine kidney, spleen, intestinal and testicle cells, and secondary cultures of embryonic bovine thymus, maintained in lactalbumin hydrolysate, Earle's balanced salt solution and ovine serum were observed for a maximum of 21 days after inoculation of E. bovis sporozoites. The sporozoites entered the cells in all of these cultures but underwent development only in primary cultures of kidney and intestinal cells and in secondary cultures of kidney, spleen, thymus, intestinal, and testicle cells. In acellular media, the sporozoites retained motility no longer than 21 hr. In the cell cultures, free motile sporozoites were seen for as long as 18 days after inoculation. Sporozoites entered cells anterior end first; the process of penetration required a few seconds to about a minute. Sporozoites were also observed leaving host cells. Intracellular sporozoites were first seen 3 min after inoculation; they were observed at various intervals up to 18 days after inoculation. In transformation of sporozoites into trophozoites a marked change in size and appearance of the nucleus took place before the change in shape of the body occurred. Trophozoites were first found 7 days after inoculation, multinucleate schizonts after 8 days, and schizonts with merozoites after 14 days. Schizonts containing merozoites were seen only in kidney, spleen, and thymus cells. The mature schizonts were smaller and represented a much lower proportion of the total number than in comparable stages of infections in calves. Schizonts with many nuclei occurred in intestinal cells; the most advanced stage seen in testicle cells was the binucleate schizont. Nuclear and cytoplasmic changes were observed in the infected cells.     

Development of Eimeria crandallis Honess from Sheep in Cultured Cells*

SYNOPSIS. Cell lines of embryonic lamb trachea (LETr), lamb thyroid (LETh), and bovine liver (BEL) as well as an established cell line of Madin-Darby bovine kidney (MDBK) were used in a study of the in vitro development of Eimeria crandallis from sheep. Excysted sporozoites were inoculated into Leighton tubes containing coverslips with monolayers of the different cell types. Coverslips were examined with phase-contrast and interference-contrast at various intervals up to 20 days after inoculation; thereafter the monolayers were fixed and stained in various ways. Freshly excysted sporozoites, with 2–10 spheroidal refractile bodies, entered all of the cell types in relatively small numbers; intracellular sporozoites were first seen 2 min after inoculation. After 24 hr, most intracellular sporozoites had only 1 or 2 refractile bodies. Before and during transformation of sporozoites, the nucleus and peripheral nucleolus increased markedly in size. Transformation resulted in usually spheroid but sometimes ellipsoid trophozoites. Trophozoites were seen first 3–4 days, and binucleate schizonts at 4–5 days after inoculation. Immature schizonts increased considerably in size and eventually had large numbers of nuclei. Some of the parasites became lobulated and the lobules often separated to form individual schizonts. In BEL, LETr and LETh cells, mature schizonts, up to 150 μm in diameter, were seen first 11–14 days after inoculation. The BEL cells were the most favorable for development. Merozoites were formed by a budding process from the surface of the schizonts as well as from blastophores. Some merozoites were seen leaving mature schizonts, but no further development was observed. Merozoites frequently were motile and had a sharply bent posterior end. Marked nuclear and cytoplasmic changes were observed in parasitized cells.     

Stages of Merogony in Multinucleate Merozoites of Eimeria magna Pérard, 1925*

SYNOPSIS. Stages of merogony of Eimeria magna were observed with the electron microscope in schizonts in ultrathin intestinal sections from white rabbits killed 4 days after inoculation. Some of the mature schizonts observed had uninucleate merozoites, whereas others had multinucleate ones. In each of the latter were rough endoplasmic reticulum, mitochondria, micronemes, refractile bodies, and occasionally, lipid droplets; some nuclei had a nucleolus. In the interior of some multinucleate merozoites, anlagen of daughter merozoites were observed. Each anlage was associated closely with a nucleus. In some of these nuclei, a cone-shaped pole of a spindle was directed toward the anlage. Each early anlage consisted of an inner membrane complex with a rhoptry anlage. A Golgi complex frequently was seen at the base of the anlage. One multinucleate merozoite, still attached to the residual body, had a merozoite anlage. In later stages of merogony, the anlagen were longer and each had a conoid. In one such merozoite, 2 merozoite anlagen were observed in close association with an eccentric intranuclear spindle, and 1 anlage had a Golgi adjunct. Another merozoite had an eccentric spindle and associated centrioles, but no visible anlagen. The finding of stages of merogony in multinucleate merozoites of E. magna indicates that these might represent a further schizogonic generation occurring in the original parasitophorous vacuole.     

Pre-Erythrocytic Development and Associated Host Responses to Haemoproteus meleagridis (Haemosporina: Haemoproteidae) in Experimentally Infected Domestic Turkeys12

ABSTRACT. Two generations of pre-erythrocytic schizogony occurred in skeletal and cardiac muscle of domestic turkeys infected with sporozoites of Haemoproteus meleagridis. First generation schizonts reached maturity approximately five days post-inoculation (DPI) and developed in capillary endothelial cells and myofibroblasts. The schizonts ranged from 12 to 20 μm in diameter and produced long (5–6 μm), slender merozoites. Early second generation schizonts were first detected in capillary endothelial cells between 5 and 8 DPI. They were cylindrical and ranged in size from 5 to 8 μm in diameter and up to 28 μm in length. Second generation schizonts which reached maturity by 17 DPI were surrounded by a thick, hyaline wall and were packed with numerous spherical merozoites less than 1 μm in diameter. Mature megaloschizonts were fusiform, ranged from 30 to 113 μm in diameter, and extended as much as 465 μm along the long axis of muscle fibers. Merozoites developed as buds from cytomeres that formed between 8 and 14 DPI. Infected turkeys developed a moderate to severe myositis within 5 DPI and were lame in one or both legs. The myositis was associated with the necrosis of scattered groups of muscle fibers. Muscle fibers surrounding mature megaloschizonts were swollen and hyaline. Megaloschizonts were surrounded occasionally by fibroblasts and infiltrates of mononuclear cells. The morphology and site of development of mature megaloschizonts of Haemoproteus meleagridis are contrasted with those of other avian haemosporidians.