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.     

Life Cycle and Host Specificity of Eimeria larimerensis Vetterling, 1964, from the Uinta Ground Squirrel Spermophilus armatus1

SYNOPSIS. Eimeria larimerensis was found in 5 species of ground squirrels and the white-tailed prairie dog. The hosts included Spermophilus armatus from Utah and Montana, S. variegatus from Utah, S. tridecemlineatus from Wyoming, S. lateralis from Utah, S. beecheyi from California and Cynomys leucurus from Wyoming. Oocysts were not present in fecal samples of S. richardconi from Montana, S. lateralis from California or S. columbianus from Washington. This coccidium could not be experimentally transmitted to S. richardsoni; however, patent infections were established in S. armatus, S. lateralis, and S. variegatus. No infections were found after inoculation of least chipmunks (Eutamius minimus), Mongolian gerbils (Meriones unguiculatus), or laboratory rats even tho excystation occurred in these animals. Resistance to infection did not develop during repeated experimental infections of S. armatus, S. lateralis, or S. variegatus. No outward signs of coccidiosis were seen in any of the experimentally infected animals. In experimentally infected S. armatus, the prepatent period was 5 days, and the patent period lasted 3–7 (mean 6.5) days. The endogenous stages were located in the epithelial cells of the jejunum and ileum. Mature 1st-generation schizonts, 1st seen 2.5 days after inoculation, contained 16–32 merozoites. Mature 2nd generation schizonts were present 3.5 days after inoculation and contained 22–46 merozoites of a larger size than those of the 1st generation. Gametocytes were 1st seen 3.5 days after inoculation and developing oocysts were present 4 days after inoculation. Macrogametes contained eosinophilic granules which coalesced to form the oocyst wall. Formation of the macrogametes took place around cytoplasmic masses within the microgametocytes.     

Development of Hepatozoon griseisciuri in Cultured Squirrel Cells*

Sporocysts of Hepatozoon griseisciuri obtained from laboratory-reared spiny rat mites (Echinolaelaps echidninus) and laboratory-reared squirrel mites (Haemogamasus reidi) were made bacteria-free and incubated in trypsin-bile for 30 min at 37 C to release sporozoites. Hepatozoon griseisciuri sporozoites were inoculated into monolayer cultures of primary adult squirrel kidney (PSK) cells and cell line cultures of neonatal squirrel kidney (SK), heart (SH), and spleen (SS) cells. Extracellular sporozoites underwent flexing, gliding, and pivoting movements similar to other coccidian sporozoites. Sporozoites entered cells in all the cultures used and were found intracellularly as early as 1 hr and as late as 10 days after inoculation. In SK, SH, and SS cells, development proceeded only to the trophozoite stage. In PSK cells, immature schizonts and mature schizonts containing 12–40 merozoites were present from 5 through 10 days after inoculation. The finding of pairs of intracellular organisms within a single parasitophorous vacuole in PSK cells suggested that endodyogeny or limited schizogony had occurred.     

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.     

Eimeria yukonensis n. sp. (Protozoa: Eimeriidae) from the Arctic Ground Squirrel Spermophilus undulatus

SYNOPSIS. Eimeria yukonensis sp. n. is described from the Arctic ground squirrel Spermophilus undulatus. The sporulated oocysts are elongate-ellipsoidal, averaging 24.5 by 13.3 μ. A micropyle is present but an oocyst residuum and polar body are absent. The ovoid sporocysts average 10.9 by 5.9 μ. The sporocyst residuum is ellipsoidal to spheroidal.     

A New Species of Eimeria from the Golden-Mantled Ground Squirrel, Spermophilus lateralis, in Northern Colorado

Oocysts of Eimeria morainensis n. sp. are described from the golden-mantled ground squirrel, Spermophilus lateralis. in Northern Colorado. The oocysts of E. morainensis are double-walled and subspherical, 20.3 × 19.8 (18.7–26.2 × 17.5–21.2) μm; and the sporocysts are ellipsoid, 12.1 × 6.9 (8.7–13.7 × 6.2–8.7) μm. Oocyst residuum and micropyle are absent, but a polar granule is present. Sporocyst residuum and Stieda body are present. Differences in oocyst characteristics provide the basis for recognition of this new species of Eimeria.     

Eimeria beldingii n. sp. and Other Coccidia (Apicomplexa) of the Ground Squirrel Spermophilus beldingi1

Fecal samples of 36 ground squirrels, Spermophilus beldingi, from Tioga Pass (elev. ca. 3315 m) in the Sierra Nevada, California, yielded oocysts of Eimeria beckeri in nine squirrels, E. citelli in four squirrels, E. beldingii n. sp. in two squirrels, and degenerated, unidentifiable oocysts in ten squirrels. Eimeria beldingii n. sp. oocysts are ellipsoidal, 30–34 × 24–30 (mean 32 × 26) μm with a two-layered, rough, striated wall, without a micropyle or residuum, with polar granules; they contain ellipsoidal or ovoid sporocysts 11–15 × 9–12 (mean 13 × 10) μm with a Stieda body and residuum.     

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.     

Eimeria lancasterensis Joseph, 1969 and E. confusa Joseph, 1969 from the Grey Squirrel Sciurus carolinensis*

SYNOPSIS. Forty grey squirrels Sciurus carolinensis were examined for coccidia during a 2-year period. Eimeria lancasterensis was found in all of them. The ellipsoidal oocysts of this species averaged 24.6 by 14.6 μ. They had no micropyle or oocyst residuum. A polar body was present. The sporocysts averaged 14.1 by 8.4 μ. The endogenous phases of the parasite were found in the epithelial cells of the villi thru the entire length of the small intestine. E. confusa was found in one of 40 squirrels. The oocysts of this species were subspherical, occasionally ellipsoidal or rarely spherical; they averaged 33.2 by 26.7 μ. Oocyst residuum and micropyle were absent. Polar granules ranged in number from 0–5. The sporocysts averaged 19.6 by 12.1 μ. The prepatent period for this species was 7–8 days and the patent period 6–13 days. E. ontarioensis was found in 3 of the 40 squirrels.     

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.     

Fire and ice: genetic structure of the Uinta ground squirrel (Spermophilus armatus) across the Yellowstone hotspot

The range of the Uinta ground squirrel, Spermophilus armatus , is centred over one of the most tectonically active regions today, the Yellowstone hotspot. We document the role of Quaternary tectonic and climatic history on the genetic structure of this species by screening museum and extant individuals throughout its range. Phylogeographic, divergence time, and demographic analyses of partial mitochondrial cytochrome b and control region DNA sequences yield insight into the cadence of evolution across three spatiotemporal scales: (i) a relatively deep intraspecific divergence of S. armatus into three lineages coincident with the last major volcanic eruption in the region and maintained by the Snake River Plain; (ii) demographic expansion in two lineages corresponding to the time of last deglaciation of the region; and (iii) a recent (< 50 years) local extinction of the third lineage coincident with climatic change and conversion of habitat for agricultural purposes in eastern Idaho. Beyond these inferences, our study highlights the unique value of museum material to phylogeography, and shows that small mammal recolonization of previously glaciated montane 'islands' differs from northward postglacial expansion observed in areas previously covered by continental ice sheets. Montane 'islands' may harbour high genetic diversity because of admixture and recurrent expansion/extinction.     

Eimeria ontarioensis n. sp., E. confusa Joseph, 1969 and Eimeria sp. (Protozoa: Eimeriidae) from the Ontario Gray Squirrel Sciurus carolinensis*

SYNOPSIS. Eimerians from 27 gray squirrels Sciurus carolinensis sampled in Waterloo County were studied. Two double-walled species, E. ontarioensis n. sp. which was piriform (36.8 × 23.5) and E. confusa Joseph which was subspherical (35.7 × 28.4), were described and compared with similar species from the genus Sciurus. Eimeria sp., ellipsoidal (24.9 × 14.4), from Ontario gray squirrels resembled E. ascotensis and E. neosciuri and was a new eimerian record from Ontario. The zoogeographic significance of E. ontarioensis n. sp. and E. sp. in North American gray squirrels is discussed.     

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.     

Natural Infection of the Ground Squirrel (Spermophilus spp.) with Echinococcus granulosus in China

Background

Echinococcus granulosus is usually transmitted between canid definitive hosts and ungulate intermediate hosts.

Methodology/Principal Findings

Lesions found in the livers of ground squirrels, Spermophilus dauricus/alashanicus, trapped in Ningxia Hui Autonomous Region, an area in China co-endemic for both E. granulosus and E. multilocularis, were subjected to molecular genotyping for Echinococcus spp. DNA. One of the lesions was shown to be caused by E. granulosus and subsequently by histology to contain viable protoscoleces.

Conclusions/Significance

This is the first report of a natural infection of the ground squirrel with E. granulosus. This does not provide definitive proof of a cycle involving ground squirrels and dogs or foxes, but it is clear that there is active E. granulosus transmission occurring in this area, despite a recent past decline in the dog population in southern Ningxia.     

The Effect of Hibernation on Lipid Metabolism in the Neocortex of the Long-Tailed Ground Squirrel Spermophilus undulatus

Biophysics - The effect of hibernation on the content of individual phospholipids and neutral lipids, as well as the effect of the states of torpor (dormant long-tailed ground squirrels...     

Fatty Acids of the Liver and the Blood Plasma During the Hibernation of the Yakutian Ground Squirrel Spermophilus undulatus

Biophysics - Abstract—We investigated the effect of hibernation on the levels of fatty acids and monoglycerides in the liver tissue and in the blood plasma of the Yakut ground squirrel...