Sarcocystis Cruzi Research Articles

In vitro Excystation of Sarcocystis capracanis, Sarcocystis cruzi and Sarcocystis tenella (Apicomplexa)

Improved rates of in vitro excystation of sporozoites from sporocysts of Sarcocystis capracanis, Sarcocystis cruzi, and Sarcocystis tenella were obtained by pretreating sporocysts with an aqueous sodium hypochlorite (NaOCl) solution followed by incubation in excysting fluid (EF). After pretreatment with NaOCl, sporocysts were washed 4 times in Hanks' balanced salt solution and then incubated in various EF (pH 7.4) at 38.5 C in 5% CO2-95% air. Maximum rates of excystation (free sporozoites/(sporozoites in sporocysts + free sporozoites) X 100) for all 3 species of Sarcocystis occurred at 4 hr after incubation in EF. These rates were 17% for S. capracanis after incubation in EF containing 2% trypsin + 10% caprine bile; 90% for S. cruzi in 2% trypsin + 10% bovine bile; and 20% for S. tenella in 2% trypsin + 10% caprine bile. Only a 40% excystation rate occurred in sporocysts of S. cruzi that had been stored previously for 14 days in aqueous potassium dichromate. Excysted sporozoites of S. capracanis, S. cruzi, and S. tenella penetrated and developed to mature meronts in bovine pulmonary artery endothelial cells or bovine monocytes.

Outbreak of sarcocytosis in dairy cattle

Sixteen of 32 Friesian calves, 8 to 10 weeks old, died over 4 weeks. The calves were housed in pens previously used by dogs. Clinical signs included anorexia, pale mucous membranes, rapid weight loss, coughing and palpably enlarged superficial lymph nodes. At necropsy, calves were emaciated and had generalised enlargement of lymph nodes, pale mottling of skeletal muscles, excess peritoneal, thoracic and pericardial fluid and subpleural and subepicardial haemorrhages. Histologically there was a lymphadenitis, myositis, myocarditis, glomerulonephritis, interstitial pneumonia and encephalitis. Schizonts of a sporozoan parasite, presumably Sarcocystis cruzi were found in the endothelial cells of blood vessels in many organs.

Perturbed metabolism and hormonal profiles in calves infected with Sarcocystis cruzi

Plasma concentrations of growth hormone (GH), thyroid stimulating hormone (TSH), insulin (IN), thyroxine (T 4), and triiodothyronine (T 3) in addition to metabolic parameters [N balance (NB), urinary 3-methylhistidine (TMH), urinary creatinine (CR), and urinary hydroxyproline (HP)] were measured in 4-mo-old Holstein steers divided equally among groups infected with Sarcocystis (I), noninfected ad libitum fed (C), and noninfected pair fed to I (PF) (7 steers per treatment). Effects of infection on these parameters beyond those attributable to altered dietary intake were determined using orthogonal contrasts (effect of intake, C vs I + PF; effect of infection, PF vs I). NB was higher in C than I and PF (P<.05) and lower in I than PF (P<.02). Hydroxyproline and CR were influenced by intake (P<.05) and HP excretion was reduced in association with infection (P<.05). Reduced intake was associated with lowered mean basal plasma concentrations of GH, IN, T 3 and T 4 (P<.05). Infection further reduced (P<.001) plasma T 3 concentration. Triiodothyronine and T 4 responses following an intravenous bolus of thyrotropin releasing hormone (TRH) were measured. The magnitude of the responses in I and PF were lower than those observed in C (P<.05). Plasma T 3 responses were further reduced in association with infection (P<.05). Insulin responses to intravenous arginine infusion (ARG) were also low in association with reduced intake. Growth hormone responses to TRH or ARG were affected by the level of feed intake only. These data suggest that hormonal perturbations associated with the insult of infection further compromise metabolism and the direction of nutrient partitioning that would ordinarily be associated with developmental growth in young steers beyond those responses anticipated from solely the reduction of feed intake.

Hematologic values in calves infected with sarcocystis cruzi

Calves were inoculated with 2 × 10 5 Sarcocystis cruzi sporocysts. Red cell mass decreased dramatically between Days 21 and 35 post-infection and plasma volume increased concurrently, so that blood volume did not change significantly. Mild reticulocytosis and increased pyrimidine 5′ nucleotidase activity in erythrocytes occurred between Days 35 and 42. Antiglobulin tests with anti-bovine IgG, IgM and C3 were negative, with the exception of a positive test for C3 in 1 of 6 infected calves.

An outbreak of sarcocystosis in dairy cattle.

Sixteen of 32 Friesian calves, 8 to 10 weeks old, died over 4 weeks. The calves were housed in pens previously used by dogs. Clinical signs included anorexia, pale mucous membranes, rapid weight loss, coughing and palpably enlarged superficial lymph nodes. At necropsy, calves were emaciated and had generalised enlargement of lymph nodes, pale mottling of skeletal muscles, excess peritoneal, thoracic and pericardial fluid and subpleural and subepicardial haemorrhages. Histologically there was a lymphadenitis, myositis, myocarditis, glomerulonephritis, interstitial pneumonitis and encephalitis. Schizonts of a sporozoan parasite, presumably Sarcocystis cruzi were found in the endothelial cells of blood vessels in many organs.

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.

Development of ox-coyote cycle of Sarcocystis cruzi.

ABSTRACTThe ox‐coyote cycle of Sarcocystis cruzi was studied by killing 38 calves between 4 and 153 days postinoculation (DPI) with 55 × 103‐5 × 108 sporocysts from the intestines of coyotes. At 4 DPI, a zoite was found within the lumen of a mesenteric lymph node artery. At 7 DPI, zoites were found in mononuclear cells and in endothelial cells in mesenteric arteries. First generation meronts (41.0 × 17.5 μm in diameter) occurred 7–26 DPI in mesenteric lymph nodes. At 19–46 DPI, second generation meronts occurred in kidneys, muscles, and other tissues: renal meronts were 19.6 × 11.0 μm, and intramuscular meronts were 25.0 × 11.1 μm. Merozoites were found in the peripheral blood 17 DPI and later at 24–46 DPI. They divided by endodyogeny in mononuclear cells. Sarcocysts were seen first in the heart at 45 DPI and contained one or two metrocytes. At 55 DPI, sarcocysts containing only metrocytes were found in striated muscles, heart, and in smooth muscles of the urinary bladder, rumen, omasum, abomasum, and small intestine. At 67, 87, 112, and 153 DPI, sarcocysts were found only in striated muscles and in the heart. At 67 DPI, sarcocysts were up to 360 μm long. They contained only metrocytes and were not infective to the dog. At 86 DPI, sarcocysts contained mostly bradyzoites, a few metrocytes, and were infective to a coyote. The thin‐walled sarcocysts grew to a maximum length of 800 μm and contained bradyzoites that were 10.9 × 3.0 μm. At 90 DPI, two mature sarcocysts were found in 2 of 73 sections of brain and spinal cord; hundreds of sarcocysts were present in sections of tongue and heart of this calf. Gametogony occurred in the small intestine of the coyote. Macro‐and microgamonts were found in goblet cells of the small intestines of coyotes 6 h after the ingestion of infected meat. Microgamonts were few and contained 3–11 slender gametes. Oocysts were seen at 12 h and sporulation was completed 9 DPI. The prepatent period in the coyote was 8 days. The ox‐coyote cycle is compared with ox‐dog cycle.

Early developmental stages of Sarcocystis cruzi in calf fed sporocysts from coyote feces.

The development of Sarcocystis cruzi was studied in an 11-day-old calf killed seven days postinoculation with 5 x 10(8) sporocysts from feces of coyotes. Uninucleate zoites were found in arteries of mesenteric lymph nodes but not in other organs. Zoites measured 4.9 x 3.0 (3.5-7.0 x 2.1-3.5) micrometers. Of the 36 zoites studied, 31 were in endothelial cells, four were in macrophages in the lumen of arteries, and one was free in the lumen of an artery. Infected endothelial cells were two to three times larger than uninfected cells. Zoites appeared structurally similar to sporozoites. The occurrence of zoites in macrophages suggests that sporozoites of Sarcocystis might use such cells to reach the site of their first merogony.

Development and ultrastructure of first-generation meronts of Sarcocystis cruzi in calves fed sporocysts from coyote feces.

The development of Sarcocystis cruzi Hasselmann (syn. S. fusiformis Railliet) meronts was studied in seven 7- to 10-day-old calves filled 4, 7, 11, 15, 22, 25 and 28 days postinoculation (DPI) with 5 x 10(7) sporocysts from feces of coyotes. No meronts were found 4 and 7 DPI. Young and intermediate meronts with 1-16 nuclei were found in endothelial cells of arteries in mesenteric lymph nodes, but not in kidneys 11 DPI. Mature meronts were noted in endothelial cells of arteries, arterioles, or capillaries of many organs of calves killed 15 to 25 DPI. No first-generation meronts were found 28 DPI. By electron microscopy, all stages of the first-generation merogony were found free within the host cell cytoplasm and not within a parasitophorous vacuole. The appearance of intranuclear spindles preceded the formation of merozoites by endopolygeny. Mature meronts measured 41.0 x 17.5 (34-50 x 15-24) microgram, contained approximately 100-350 merozoites, and had 2 to 4 relatively small residual bodies, 2.8 microgram in diameter. Merozoites measured 6.3 x 1.5 (5.5-7 x 1 microgram) and contained most of the organelles characteristically found in coccidian merozoites. Micropores were observed in merozoites, but not in young and intermediate meronts. Merozoites were seen free in the lumen of blood vessels, in intracellular areas, and free within the host cell cytoplasm.

Pathophysiological changes in urine and blood from calves experimentally infected with Sarcocystit cruzi.

Two 8-week experiments were conducted to determine the relationships between nutritional stress and pathophysiological changes in male Holstein calves infected with Sarcocystis cruzi. Calves were infected by oral inoculation with 200 000 S. cruzi sporocysts. In the first experiment weight gain reduction was greatest in inoculated calves during weeks 4 and 5 after inoculation. Feed intake was reduced during the 5th week. Erythrocyte count was reduced during week 5 and haemoglobin was reduced during week 6. The 24-h excretion of urinary and urea nitrogen from the inoculated calves was increased by treatment. In the 2nd experiments, both the feed-restricted and inoculated calves lost weight during weeks 4 and 5; feed intake was lower from week 5 to 8 inclusive. Urine volume from inoculated calves was lower during week 8. Lower urine excretions of sodium and potassium resulted from S. cruzi inoculation. There was a non-significant trend for higher urinary zinc excretion in the inoculated group during week 4. Urine nitrogen excretion from inoculated calves was higher during week 6. The urinary excretion of 3-methylhistidine from the inoculated calves was higher during week 4 and excretion of guanine was higher during weeks 4, 5 and 8. S. cruzi has several specific pathophysiological effects on calves beyond those induced by nutritional stress.

Fine Structure of Immature Cysts of Sarcocystis cruzi

Sarcocystis cruzi forms cysts in striated muscle of the bovine host following schizogony. The fine structure of the immature cyst within muscle fibers of the ventricular myocardium was studied in relation to its development and to the multiplication of parasites within it. The young cyst is enclosed by a cyst wall containing numerous small protuberances. Metrocytes within the cyst are irregular in shape and are separated from each other and the cyst wall by a thin layer of ground substance. The parasite multiplies by endodyogeny within the metrocyte. As the cyst enlarges, the host muscle fiber is disrupted and large protrusions are present in the cyst wall.

The prevalence of Sarcocystis spp in dogs and red foxes

Protozoan parasites of the genus Sarcocystis have been recognised for many years as intramuscular cysts of numerous vertebrates. It is only comparatively recently that the two-host nature of the life cycle has been recognised and that the intramuscular cysts are a stage in the developmental cycle of coccidian parasites of flesh eating mammals (Fayer 1974, Fayer and Johnson 1973, 1974, Rommel and others 1972, Dubey 1976). Carnivores ingest the intramuscular cysts from herbivores and presumably from other animals too and eventually shed sporulated tetrazoic sporocysts in their faeces. The cystic stages which occur in the flesh of herbivores are probably non-pathogenic but the earlier stages in which schizonts develop in vascular endothelium may be severely pathogenic. Sarcocystis cruzi, S ovicanis and S porcifelis are known to be severely pathogenic in cattle, sheep and pigs respectively (Dubey 1976). Observations on the prevalence of Sarcocystis spp in the faeces of working farm dogs, greyhounds and foxes (Vulpes vulpes) are recorded.