Sarcocystis hemionilatrantis (Sp. N.) life cycle in mule deer and coyotes.

Fifteen coyotes (Canis latrans) shed sporulated sporocysts in their feces after eating freshly ground skeletal muscles from a mule deer (Odocoileus hemionus hemionus) infected with microscopic-sized cysts of Sarcocystis. Sporocysts were shed intermittently from 12 to 36 days after ingestion of the infected meat. Sporocyst size averaged 14.4 X 9.3 mum. Eleven mule deer fawns orally inoculated with these sporocysts became infected and 9 of 11 died between post-inoculation days (PID) 27 and 63. Clinical signs of anorexia, weight loss, pyrexia and weakness were evident prior to death. A calf (Bos taurus) and two lambs (Ovis aries) orally inoculated with these sporocysts did not become infected and remained healthy throughout the experiments. Similarly, uninoculated control animals consisting of three mule deer fawns, two lambs and one calf remained healthy during the experiment. Preliminary histologic examinations conducted on selected tissues from all animals revealed microscopic-sized schizogonous stages in macrophages, between muscle fibers and near blood vessels in the esophagus, heart, biceps femoris, semi-membranosus, diaphragm and tongue from seven of eight fawns which died between PID 27 and 39. Developing or mature muscle cysts were not found in fawn tissue until PID 60. Sarcocysts were found in the three infected fawns examined after this time. Muscle cysts or earlier schizont stages were not found in tissues from the inoculated or uninoculated calves and lambs. A single muscle cyst was found in one control fawn; the other two control fawns were negative for both muscle cysts and other schizogonous stages. These results established that the life cycle of this species of Sarcocystis can be completed with coyotes as the definitive host and mule deer as the intermediate host. Based on the demonstrated host specificity and earlier findings, the name Sarcocystis hemionilatrantis is proposed for this parasite of mule deer and coyotes.     

Development of Sarcocystis fayeri in the equine

Eight ponies and a horse were inoculated orally with sporocysts of Sarcocystis fayeri from dogs. They were examined for clinical signs of infection and killed 10, 15, 20, 25, 30, 50 (horse), 77, 101, and 156 days after inoculation (DAI). Elevated temperature was observed in three ponies 20 and 26 DAI and anemia was observed in three ponies and the horse 15 to 69 DAI. Schizonts were found in or near cells lining capillaries or arteries of the heart, brain, and kidney 10, 20, and 25 DAI. Immature cysts containing only metrocytes were first found in muscles 50 DAI. Mature intramuscular cysts containing metrocytes and zoites or zoites alone were found 77, 101, and 156 DAI and produced patent infections in dogs fed infected meat.     

Experimental transmission of a sarcosporidian from Alpine ibex to domestic sheep and goats

Sporocysts of Sarcocystis sp. from dogs fed with ibex meat were orally inoculated into kids and lambs. Three kids, given 4 x 10(6) and 4 x 10(4) sporocysts, respectively, died from acute sarcocystosis. Schizonts, though found in all the tissues of these kids, were particularly numerous in the kidneys, brain and spinal cord. Another three kids inoculated with 5 x 10(3) sporocysts and two lambs, inoculated with 1 x 10(6) and 5 x 10(3) sporocysts, respectively, showed no clinical signs and were sacrificed between 111 and 130 days after infection. Mature sarcocysts were found both in the heart and striated muscles of these animals. No parasitic stage was found in two kids and two lambs used as uninoculated controls. Biological differences between Sarcocystis sp. from ibex and the other sarcosporidians with a canine-caprine or canine-ovine cycle are stressed.     

Characterization of an unidentified Sarcocystis falcatula-like parasite from the South American opossum, Didelphis albiventris from Brazil

An unidentified isolate of a Sarcocystis falcatula-like parasite was obtained from the lungs of budgerigars (Melopsittacus undulatus) fed sporocysts from a naturally-infected South American opossum, Didelphis albiventris from Brazil. Four captive budgerigars fed sporocysts from the opossum intestine died of acute sarcocystosis 8, 10, and 12 days after oral inoculation (DAI); one budgerigar was killed 12 DAI when it was lethargic. Schizonts and merozoites found in the lungs of the budgerigars reacted mildly with polyclonal S. falcatula antibody. The parasite was isolated in equine kidney cell cultures inoculated with lung tissue from a budgerigar that was killed 12 DAI. Two budgerigars inoculated subcutaneously with 100,000 culture-derived S. falcatula merozoites developed acute sarcocystosis and S. falcatula-like schizonts were found in their lungs 15 and 16 DAI. Four budgerigars kept as unfed controls in the same environment remained free of Sarcocystis infection. The parasite underwent schizogony in African green monkey kidney cells and bovine turbinate cells. Merozoites divided by endopolygeny, often leaving a residual body. Polymerase chain reaction studies using primers JNB33/JNB54 and Hinf I and Dra I digestion indicated that the isolate was not S. falcatula. Results of this study indicated that the South American opossum, D. albiventris, is a definitive host for yet another S. falcatula-like parasite.     

Protective Immunity to Sarcocystis capracanis-Induced Abortion in Dairy Goats1

ABSTRACT. Eleven female goats (Nos. 1 to 11) were each inoculated orally with 10⁴ sporocysts of Sarcocystis capracanis, and four female goats (Nos. 12 to 15) were not inoculated. Between 31 and 69 days after inoculation (DAI) goats were mated with a single buck; one goat (No. 5) did not breed. Eight inoculated goats were challenged with 10⁵or 10⁶ sporocysts, 135 DAI. Two of four goats challenged with 10⁶ sporocysts and one of three goats challenged with 10⁵ sporocysts aborted one month before the expected time of parturition. The three inoculated goats that were not challenged delivered healthy kids. All inoculated goats including the nonpregnant one (No. 5) were only mildly ill from the primary or challenge inoculations. Two of the four control goats challenged with 5 × 10⁴ or 10⁵ sporocysts aborted 21 days later, and both died of sarcocystosis 25 and 88 DAI. The two remaining control goats delivered normal kids. The results indicate that immunization prior to pregnancy protects some but not all goats from .Sarc0c.es/is-induced abortion.     

Experimental infections of Sarcocystis cruzi, Sarcocystis tenella, Sarcocystis capracanis and Toxoplasma gondii in red foxes (Vulpes vulpes)

Four littermate 6-wk-old red foxes (Nos. 1-4) were fed Toxoplasma gondii, Sarcocystis cruzi, S. tenella and S. capracanis. One littermate fox (No. 5) served as the control. Two foxes (Nos. 1, 2) were fed tissue cysts of T. gondii and two foxes (Nos. 3, 4) were fed oocysts of T. gondii. Twenty-one to 42 days later, the same five foxes were used to test the infectivity of meat of goat, sheep, and ox experimentally inoculated with Sarcocystis. Fox 2 was fed goat meat and shed S. capracanis-like sporocysts 10 days later. Foxes 3 and 4 were fed beef, and they shed S. cruzi-like sporocysts 9 days later. Fox 5 was fed sheep meat and shed S. tenella-like sporocysts 8 days later. Foxes were killed between 36 and 55 days of the experiment and their tissues were inoculated into mice to recover T. gondii. All foxes remained clinically normal and T. gondii was recovered from all inoculated foxes and not from the control. Sarcocystis sporocysts from foxes induced lethal infections in goats, sheep, and ox. The sporocysts, meronts, merozoites, and sarcocysts of fox-derived parasites were similar to those derived from coyotes or dogs. It was concluded that the red fox can act as a final host for the three pathogenic species of Sarcocystis in cattle, sheep, and goats.     

The gamma interferon knockout mouse model for sarcocystis neurona: comparison of infectivity of sporocysts and merozoites and routes of inoculation.

The dose-related infectivity of Sarcocystis neurona sporocysts and merozoites of 2 recent isolates of S. neurona was compared in gamma interferon knockout (KO) mice. Tenfold dilutions of sporocysts or merozoites were bioassayed in mice, cell culture, or both. All 8 mice, fed 1,000 sporocysts, developed neurological signs with demonstrable S. neurona in their tissues. Of 24 mice fed low numbers of sporocysts (100, 10, 1), 18 became ill by 4 wk postinoculation, and S. neurona was demonstrated in their brains; antibodies (S. neurona agglutination test) to S. neurona and S. neurona parasites were not found in tissues of the 6 mice that were fed sporocysts and survived for >39 days. One thousand culture-derived merozoites of these 2 isolates were pathogenic to all 8 mice inoculated subcutaneously (s.c.). Of the 24 mice inoculated s.c. with merozoites numbering 100, 10, or 1, only 3 mice had demonstrable S. neurona infection; antibodies to S. neurona were not found in the 21 mice that had no demonstrable organisms. As few as 10 merozoites were infective for cell cultures. These results demonstrate that at least 1,000 merozoites are needed to cause disease in KO mice. Sarcocystis neurona sporocysts were infective to mice by the s.c. route.     

Sarcocystis transmitted by blood transfusion.

Merozoites were found in blood smears from calves, lambs, and pigs exhibiting signs of acute sarcocystosis after oral infection with sporocysts of Sarcocystis bovicanis, Sarcocystis ovicanis, and Sarcocystis suihominis. In each of 3 experiments, whole blood containing merozoites was transfused from an infected host to 2 uninfected recipients of the same species; 2 additional animals of the same species served as uninfected nontransfused controls. The bovine, ovine, and porcine donors all died of acute sarcocystosis. Clinical signs of sarcocystosis were not seen in any transfusion recipients or controls. All recipients and controls were killed and histologic specimens of esophagus, diaphragm, heart, skeletal muscle, and tongue were examined microscopically. Large numbers of intramuscular cysts were present in transfusion recipients, whereas few or no cysts were present in controls, indicating that S. bovicanis, S. ovicanis, and S. suihominis had been transmitted between intermediate hosts of the same species by blood transfusion.     

Experimental transmission of Sarcocystis speeri Dubey and Lindsay, 1999 from the South American opossum (Didelphis albiventris) to the North American opossum (Didelphis virginiana)

Sarcocystis speeri Dubey and Lindsay, 1999 from the South American opossum Didelphis albiventris was successfully transmitted to the North American opossum Didelphis virginiana. Sporocysts from a naturally infected D. albiventris from Argentina were fed to 2 gamma-interferon knockout (KO) mice. The mice were killed 64 and 71 days after sporocyst feeding (DAF). Muscles containing sarcocysts from the KO mouse killed 71 DAF were fed to a captive D. virginiana; this opossum shed sporocysts 11 days after ingesting sarcocysts. Sporocysts from D. virginiana were fed to 9 KO mice and 4 budgerigars (Melopsittacus undulatus). Schizonts, sarcocysts, or both of S. speeri were found in tissues of all 7 KO mice killed 29-85 DAF; 2 mice died 39 and 48 DAF were not necropsied. Sarcocystis stages were not found in tissues of the 4 budgerigars fed S. speeri sporocysts and killed 35 DAE These results indicate that S. speeri is distinct from Sarcocystis falcatula and Sarcocystis neurona, and that S. speeri is present in both D. albiventris and D. virginiana.     

The camel (Camelus dromedarius) as an intermediate host for Hammondia heydorni

Dogs fed raw camel meat containing two types of cysts shed unsporulated Hammondia heydorni oocysts and later sporulated Sarcocystis sporocysts in their feces, but were resistant to reinfection with the Hammondia cysts. Sporulated H. heydorni occysts did not induce an enteroepithelial cycle in dogs, but resulted in the formation of muscle cysts.     

Experimental Sarcocystis ovicanis infection in lambs: salinomycin chemoprophylaxis and protective immunity

Salinomycin, administered prophylactically, was effective in controlling clinical sarcocystosis resulting from infection with Sarcocystis ovicanis in two experiments involving a total of 50 lambs. In each experiment, five lambs were used in each of five test groups--uninoculated and unmedicated, inoculated and unmedicated, uninoculated and medicated (2 mg/kg), inoculated and medicated (2 mg/kg), and inoculated and medicated (1 mg/kg). Salinomycin was included in the feed of each medicated group for 30 days beginning on the day of oral inoculation with 10(6) S. ovicanis sporocysts. Lambs were challenged with 10(6) sporocysts 9 wk PI and the experiment was terminated 7 wk later. Data from deaths, weight gains, body temperatures, packed cell volumes, hemoglobin values, serum protein levels, and postmortem and histological examinations indicated that salinomycin at both doses tested reduced the number of deaths and severity of signs of sarcocystosis in infected lambs as compared with unmedicated controls. Infected, medicated lambs developed a protective immunity as determined by challenge inoculation. An additional group of five lambs, each inoculated orally with 10(6) sporocysts, were treated therapeutically with 2 mg/kg per day beginning 21 days after inoculation. All five died from acute sarcocystosis.     

The ferret (Putorius putorius furo), an additional final host of Sarcocystis muris (author's transl)]

Two ferrets were fed mice experimentally infected with Sarcocystis muris. After 7 days they excreted with their faeces for 9 days sporocysts which were morphologically indistinguishable from S. muris sporocysts. Five mice which each received 70 of these sporocysts orally developed macroscopically visible cysts of S. muris in their musculature after 4 months.     

The asexual pre-cyst development of Sarcocystis tenella in experimentally infected specific pathogen-free lambs

Twenty-two lambs raised under sporozoa-free conditions were infected at weaning with 0.25–2.0 million Sarcocystis tenella sporocysts obtained from dogs and then necropsied at intervals between 1.3 h and 41 days post-inoculation (dpi). Fixed and frozen sections from 47 different tissues from each lamb were examined respectively by a variety of histochemical stains and by indirect fluorescent-antibody staining. The remnants of excysted sporocysts were found within the gastro-intestinal tract between 1.3 h and 3 dpi. No enteric proliferative stage of the parasite was detected. First-generation meronts (schizonts) were detected from 6 to 19 dpi within the endothelial cells of arterioles in most organs and tissues. The meronts were undifferentiated from 6 to 9 dpi but were mature in appearance from 12 to 19 dpi (measuring 22.3 × 13.4 μm and containing 18–28 merozoites). Second-generation meronts were detected from 21 to 34 dpi within the endothelial cells of capillaries throughout the entire body. They were undifferentiated from 21 to 23 dpi but appeared mature from 25 to 34 dpi (measuring 15.2 × 10.6 μm and containing 18–38 merozoites). Several smaller meronts were then detected at 36 dpi within cells of the mononuclear phagocytic system. They were all mature in appearance (measuring 7.4 × 5.1 μm and containing 6–9 merozoites) and are thought to have formed facultatively following the incorporation of some second-generation merozoites into phagocytic cells. Only developing cysts were then detected at 41 dpi throughout the cardiac and skeletal musculature. Elements in both first- and second-generation merozoites stained markedly with haematoxylin and eosin, periodic acid-Schiff's, alkaline Giemsa-colophonium and basic fuchsin stains. However, only second-generation meronts and developing cysts exhibited strong specific reactions with the fluorescent-antibody stain.     

Sarcocystis fayeri sp. n. from the horse.

Hearts, diaphragms, esophagi, and spinal cords from 266 horses were obtained at slaughter in Creston, Ohio. Tissues were examined microscopically for Sarcocystis in sections, digested in trypsin to obtain bradyzoites, and fed to 10 dogs and 10 cats. Intramuscular cysts were found in selections of two hearts from 57 horses and four esophagi from 107 horses. The cysts were up to 900 micron long and up to 70 micron wide. The cyst wall was 1 to 2 micron thick and cross-striated. The enclosed bradyzoites were banana-shaped, 15 to 20 by 20 to 3 micron, and contained several PAS-positive granules. Bradyzoites were found in trypsin digests of seven of 57 (13%) equine tissues (heart, diaphragm, esophagus but not spinal cord) in one experiment and 10 of 47 (21%) esophagi, eight of 47 (17%) diaphragms but none of 47 hearts and spinal cords in another experiment. All of 10 dogs shed sporulated sporocysts or oocysts in feces 12 to 15 days (12 in one, 13 in eight, and 15 days in one) after digesting tissues from 169 horses. The sporocysts were 11 to 13 (12.0 +/- 0.5) by 7 to 8.5 (7.9 +/- 0.5) micron. In histologic sections of canine small intestine the sporocysts were located in the lamina propria near the tips of the villi. The 10 cats fed tissues from 266 horses did not shed Sarcocystis. A new name, S. fayeri, is proposed for this organism. Sarcocystis fayeri sporocysts (12 by 8 micron) are shorter than those of S. betrami (15 by 10 micron), the other species of Sarcocystis from the horse. The prepatent period is 12 to 15 days for S. fayeri and 8 days for S. bertrami (synonym S. equicanis Rommel and Geisel 1975).     

Failure of Development of the Sexual Phase of Eimeria intricata in Heavily Inoculated Sheep*

SYNOPSIS Each of 4 bottle-fed lambs received totals of 3500-8100 sporulated oocysts of Eimeria intricata in series of 3-6 inoculations during 4- to 14-day periods; all produced oocysts of the parasite. The periods of patency in some infections indicated that only the earliest inoculations of a series may have produced oocysts. Fifteen other bottle-fed lambs were inoculated, each with a single dose of 0.01-2 million sporulated oocysts, and necropsied at intervals of 1-2 days over 29 days. Parasites apparently resulting from the inoculations were found in only the lambs killed up to and including 17 days postinoculation; the most advanced stages were 2nd generation schizonts. It was concluded that a reaction occurred against the large numbers of asexual stages of E. intricata, which were evidently present, and the infections were terminated before formation of sexual stages.     

Parasitemia and early tissue localization of Sarcocystis neurona in interferon gamma gene knockout mice fed sporocysts.

Early localization and parasitemia of Sarcocystis neurona were studied in gamma interferon gene knockout (KO) mice fed S. neurona sporocysts. Mice were examined for S. neurona infection histologically and immunohistochemically and by bioassay in KO mice. For bioassay, blood and tissue homogenates were inoculated subcutaneously into KO mice. Parasitemia was demonstrated by bioassay in KO mice 1-8 days after feeding sporocysts (DAFS). Sporozoites were seen in histologic sections of all regions of the small intestine and in cells in Peyer's patches of a mouse killed 6 hr after feeding sporocysts. At 1 DAFS, organisms were present in all regions of the small intestine and were also seen in mesenteric lymph nodes. At 3 DAFS, organisms had begun to invade extraintestinal tissues. Sarcocystis neurona was demonstrated histologically in mouse brain as early as 4 DAFS.     

Observations on the endogenous stages of Eimeria crandallis in domestic lambs (Ovis aries)

Lambs reared coccidia-free were inoculated orally with various numbers of sporulated oocysts of E. crandallis and were killed between 1 and 22 days after inoculation; tissues were examined histologically. Sporozoites were seen 1, 2 and 3 days after inoculation (DAI) in crypt epithelial cells in the mid-jejunum. Infected cells migrated into the lamina propria where the parasite within them developed into a firstgeneration meront containing about 250,000 merozoites at 10 DAI. A second generation of meronts was seen at 10–12 DAI, each containing up to about 10 merozoites, situated mainly at the bases of crypts in the jejunum and ileum but also in the caecum. From 11 DAI pro-gamonts were seen which were enveloped by the host cell nucleus and which divided in synchrony with the host cell for an undetermined number of generations. Mature gamonts began to develop from them by 16 DAI. Oocyst output began at 16 DAI and rose to a peak at about 22 DAI. Up to 10⁸ oocysts were produced per oocyst inoculated. They showed wide variation in size and colour.     

Observations on the endogenous stages of Eimeria crandallis in domestic lambs (Ovis aries)

Lambs reared coccidia-free were inoculated orally with various numbers of sporulated oocysts of E. crandallis and were killed between 1 and 22 days after inoculation; tissues were examined histologically. Sporozoites were seen 1, 2 and 3 days after inoculation (DAI) in crypt epithelial cells in the mid-jejunum. Infected cells migrated into the lamina propria where the parasite within them developed into a firstgeneration meront containing about 250,000 merozoites at 10 DAI. A second generation of meronts was seen at 10–12 DAI, each containing up to about 10 merozoites, situated mainly at the bases of crypts in the jejunum and ileum but also in the caecum. From 11 DAI pro-gamonts were seen which were enveloped by the host cell nucleus and which divided in synchrony with the host cell for an undetermined number of generations. Mature gamonts began to develop from them by 16 DAI. Oocyst output began at 16 DAI and rose to a peak at about 22 DAI. Up to 10⁸ oocysts were produced per oocyst inoculated. They showed wide variation in size and colour.     

Infectivity of sarcocystis spp. from bison, elk, moose, and cattle for cattle via sporocysts from coyotes

Bison bison (bison), Cervus canadensis (elk), Alces alces (moose), and Bos taurus (cattle) musculature containing Sarcocystis spp. cysts was fed to laboratory raised Canis latrans (coyotes), Sporocysts collected from the feces of coyotes fed musculature of each of the ruminant species were fed to four groups of three laboratory-raised domestic calves, respectively, to determine if Sarcocystis spp. was transmissible from wild to domestic ruminants and if so, to compare clinical signs of infection and morphologic features of cysts with those resulting from infection with Sarcocystis bovicanis. All calves fed sporocysts of Sarcocystis from coyotes that ate bison or cattle muscle had similar clinical signs and harbored morphologically similar parasites, suggesting that both bison and cattle are intermediate hosts for S. bovicanis and that this species is transmissible between the two ruminant species. All calves fed sporocysts from coyotes that ate elk muscle or moose muscle remained asymptomatic but one calf in each group had intramuscular cysts. The finding of relatively large numbers of intramuscular cysts in one calf fed sporocysts of elk origin and smaller numbers in one calf fed sporocysts of moose origin could represent either spurious natural infections or indicate low infectivity of Sarcocystis spp. from elk and moose to cattle.