A light microscopic comparison of the cysts of four species of Sarcocystis infecting the domestic reindeer (Rangifer tarandus) in northern Norway

Fresh preparations of micro-isolated sarcocysts from skeletal and cardiac muscle of 12 reindeer were examined by light microscopy. On the basis of cyst structure and cyst wall structure 4 Sarcocystis spp. could be differentiated. New names have been proposed for 2 previously unnamed Sarcocystis spp. of reindeer, and S. grueneri has been redefined.S. rangiferi n. sp. had macroscopic cysts in skeletal muscle measuring 2106×403 µm. The cyst wall protrusions were finger-like and measured 13.2×6.7 µm. The cysts were surrounded by a layer of fibrillar material.S. tarandi n. sp. had micro- to macroscopic cysts primarily in skeletal muscle, but a few cysts were found in the heart of one animal. In skeletal muscle the cysts measured 999×75µm; in the heart the cysts were shorter and wider. The cyst wall protrusions were fingerlike and measured 9.2×2.2 µm.S. grueneri had micro- to macroscopic cysts in cardiac muscle measuring 581×137 µm. The cyst wall was thin and relatively smooth with no visible protrusions.Sarcocystis sp. had micro- to macroscopic, slender cysts in skeletal muscle measuring 916×64 µm. The cyst wall had tightly packed, short, knob-like protrusions. The cysts of this species were previously classified as cysts of S. grueneri.     

First isolation of Sarcocystis hominis from cattle in Japan.

Sarcocystis hominis was first isolated from slaughtered cattle raised in Japan. Cysts were 1,220-4,460 x 80-384 microns in size and their wall was 3 to 6 microns thick and appeared radially striated in the histopathological sections because of the presence of palisade-like villar protrusions on the surface. The protrusions were 3.1-4.3 x 0.7-1.1 microns in size and had many microtubules in the core. Two cynomolgus monkeys, Macaca fascicularis, fed with the Sarcocystis cysts began to pass sporocysts, which measured a size of 14.3-15 x 9.5-10 microns, in the feces 10 days after ingestion.     

驴肉孢子虫包囊的超微结构及其实验感染

对昆明地区的26头驴进行了肌肉检查，在24头驴肌肉中发现了肉孢子虫包囊，其然自感染率为92．3％，光镜下包囊呈梭形，具有锥状和棍棒状的突起，透射电镜下突起内微管在顶端松散排列，向下延伸集结成束，斜伸入基质层，有的和母细胞的膜相连，用含有包囊的驴肉实验感染犬和猫各2只，感染后在犬粪便中发现孢子囊和卵囊，潜隐期为11－13 d，剖检在2只犬的小肠固有层中查到孢子囊和卵囊，猫粪便中一直未检到孢子囊和卵囊，剖检亦未发现，表明驴肉孢了虫的终末宿主要是犬，而是猫，研究认为驴全内的肉孢子虫仅有1种，鉴定为柏氏肉孢子虫（Sarcocystis bertrami).     

Sarcocystis infection in wild reindeer (Rangifer tarandus) from Hardangervidda in southern Norway: with a description of the cysts of Sarcocystis hardangeri n. sp

Fresh preparations of micro-isolated sarcosysts from skeletal muscle of 5 wild reindeer were examined by light microscopy. Slender, spindelshaped cysts measuring 821 × 60 µm, and having short knob-like cyst wall protrusions were found in all animals. In 1 animal cysts different in structure from the cysts of the 4 previously known Sarcocystis spp. of reindeer were found, These cysts are considered to be cysts of a new Sarcocystis sp. of reindeer, for which the name Sarcocystis hardangeri has been proposed.S. hardangeri n. sp. had macroscopic, ovoid to cylindrical cysts measuring 1667 (900–2570) × 819 (450–1575) µm. The cysts were surrounded by a 8–10 µm thick layer of fibrillar material. After removal of this layer, relatively few and irregularly spaced, slanting protrusions became visible. The 20–30 µm long protrusions were tongue-like, and were lying close to the surface of the cyst.Cysts of S. grueneri, S. rangiferi and S. tarandi were not demonstrated in the 5 wild reindeer examined.     

The prevalence and identity of Sarcocystis in beef cattle in New Zealand

Muscle tissue from the oesophagus and diaphragm of 500 beef cattle slaughtered in New Zealand was examined for Sarcocystis infection by microscopic examination of cysts isolated from muscle samples. All cattle were infected with Sarcocystis; based on light microscopy of cysts, 98% had thin-walled Sarcocystis cruzi cysts and 79.8% had thick-walled (Sarcocystis hirsuta/Sarcocystis hominis) cysts. Cysts were also collected for electron microscopy and transmission experiments. Thick-walled cysts could not be distinguished as S. hirsuta or S. hominis by light or electron microscopy. Thick-walled cysts were fed to three cats and one human volunteer; one cat shed sporocysts but not the human volunteer. Electron microscopy of the cysts revealed many features that have not been described previously.     

Sarcocystis sp. from cattle slaughtered in Japan

Sarcocystis sp. was detected from cattle slaughtered in Saitama Prefecture, Japan. The cysts were 3,400-4,400 x 198-238 microm in size and had the thick cyst wall which was 7 to 10 microm thick and provided with finger-like villar protrusions. The protrusions were 8-9.5 x 2-2.5 microm in size and had microtubules in the core.     

The structure and identity of macroscopically visible Sarcocystis cysts in cattle

Macroscopically visible Sarcocystis spp. cysts isolated from the skeletal muscle of slaughtered cattle were examined by light- and electronmicroscopy. Transmission experiments involving cats, dogs and a human volunteer were also carried out. The cysts could only be transmitted to cats which establishes them with a high degree of certainty as Sarcocystis hirsuta. The cyst wall (including protrusions) ranged from 3.3 to 7.0 micron in thickness and the individual cyst wall protrusions from 1.2 to 2.6 micron in width. Transmission and scanning electronmicroscopy revealed previously undescribed features of the cyst wall. It appears that, with increasing age, the cyst wall protrusions become larger and develop a highly irregular surface. Their attachments to the cyst wall are slender and widely spaced indicating that growth of the cyst continues without the formation of new protrusions. Within the protrusions the fibrils become disorganised and numerous osmiophilic granules appear. It is evident that major changes in the structure of sarcocysts can occur with age.     

Sarcocystis hardangeri and Sarcocystis rangi n. sp. from the domestic reindeer (Rangifer tarandus) in northern Norway

Fresh preparations of microisolated sarcocysts from striated muscle of several domestic reindeer from northern Norway were examined by light microscopy. In cardiac muscle, cysts of S. grueneri were found. In skeletal muscle, cysts of S. rangiferi, S. tarandi and S. tarandivulpes were found in all samples examined. In the abdominal muscles of some reindeer, one or two other types of cysts were found.Cysts of one type were macroscopic in size, and ovoid to cylindrical in shape. The cysts were surrounded by a 8–12 µm thick layer of fibrous material, and measured 1682×910 µm. The cysts had relatively few and irregularly distributed, 20–35 µm long, and 3–5 µm wide, linguiform cyst wall protrusions, which could only be seen after removal of the fibrous layer. These cysts were classified as cysts of S. hardangeri, a species previously described from wild reindeer in southern Norway.Cysts of the other type were long and slender, measuring 5460–12700 (8994 ± 2575) × 95–280 (180 ± 50) µm. The cysts had numerous very fine, flexible, hair-like cyst wall protrusions, which were 8–10 [xm long and less than 0.5 µm thick. These cysts are considered to belong to a new Sarcocystis species of reindeer, for which the name Sarcocystis rangi n, sp. is proposed. The reindeer is recorded as the intermediate host for 6 different species of Sarcocystis.     

The fox as definitive host for Sarcocystis sp. Gjerde, 1984 from skeletal muscle of reindeer (Rangifer tarandus). With a proposal for Sarcocystis tarandivulpes n. sp. as replacement name

Skeletal muscle of 5 wild reindeer was examined for sarcocysts and used for experimental infection of 6 foxes. Skeletal and cardiac muscle of another reindeer were only examined for sarcocysts. The skeletal muscle of all animals was infected with Sarcocystis sp.. In 2 of the animals cysts of S. hardangeri were also present. The single heart examined contained only cysts of S. grueneri.Four foxes given skeletal muscle containing apparently only cysts of Sarcocystis sp., started shedding Sarcocystis sporocysts, measuring on average 13.6×9.8 µm, after a prepatent period of 10–12 days. Two foxes given skeletal muscle containing cysts of both Sarcocystis sp. and S. hardangeri shed similar sporocysts, measuring on average 13.5×9.7 µm, after a prepatent period of 10–12 days.Based on the results from the present and previous investigations, Sarcocystis sp. is considered to have foxes (Vulpes vulpes and Alopex lagopus) and dogs (Ganis familiaris) as definitive hosts, becoming the second species of Sarcocystis with a known reindeer/Canidae life cycle. The name Sarcocystis tarandivulpes n. sp. is proposed as a replacement name for Sarcocystis sp. Gjerde, 1984 from skeletal muscle of reindeer.     

Ultrastructure of the cysts of Sarcocystis tarandivulpes from skeletal muscle of reindeer (Rangifer tarandus tarandus)

The cysts of S. tarandivulpes were found to be limited by a unit membrane which has been called the cyst membrane. The surface of the cysts was covered by closely packed and hexagonally arranged knob-like protrusions. The protrusions were 0.6–1.2 μm long and had an elliptical cross section. At the base of and between the bases of the protrusions the cyst mem brane was raised into low anastomosing folds which delineated shallow compartments. Between the folds the cyst membrane formed small vesicle-like invaginations into the cyst. On the apical part of the protrusions the cyst membrane had a smooth contour and was underlined by 2 layers of electron-dense material. Cyst ground substance divided the interior of the cyst into compartments containing either metrocytes or cystozoites. Cystozoites undergoing endodyogeny were present among the nondividing cystozoites. Some new terms were introduced to denote structures at the border of the cyst. The old terms are reviewed and the structural resemblance between S. tarandivulpes and S. odocoileocanis from Odocoileus virginianus is discussed.     

Sarcocystis infection in the white-tailed deer (Odocoileus virginianus) in Montana: intensity and description of Sarcocystis odoi n sp

Sarcocystis infection was diagnosed in 27 of 36 (75%) samples of meat from white-tailed deer (Odocoileus virginianus) in Montana. Two structurally distinct thin- and thick-walled sarcocysts were found in the white-tailed deer; the thin-walled sarcocysts were those of S odocoileocanis. A new name, S odoi, was proposed for the thick-walled sarcocysts. Sarcocysts of S odoi were up to 1,050 microns long and 260 microns wide and contained a 6.5- to 12-microns thick wall. The ultrastructure of sarcocysts of S odocoileocanis was compared with that of S odoi. A cat fed meat containing thin- and thick-walled sarcocysts shed oocysts and sporocysts 24 days later. The sporocysts in the cat's feces were 13.3 X 9.6 microns and probably belonged to S odoi.     

The raccoon dog (Nyctereutes procyonoides) as definitive host for Sarcocystis spp. of reindeer (Rangifer tarandus)

A raccoon dog (Nyctereutes procyonoides; Family: Canidae), was given cardiac muscle of reindeer infected with S. grueneri, and started shedding Sarcocystis sporocysts 10 days post feeding. The sporocysts measured 13.9 (12.4–15.7) × 10.1 (9.2–11.2) µm, and were excreted for at least 16 days. The raccoon dog is thus an additional definitive host for S. grueneri (Yakimoff & Sokoloff, 1934) Gjerde, 1984.Another raccoon dog was given skeletal muscle infected with 4 species of Sarcocystis, none of which was S. grueneri. The raccoon dog started shedding Sarcocystis sporocysts on day 10 post feeding, and excreted sporocysts for at least 16 days. The sporocysts measured 14.0 (12.3–15.6) × 10.1 (9.2–11.2) µm, and are considered to be sporocysts of S. tarandivulpes Gjerde, 1984.This is the first record of the raccoon dog as an experimental definitive host for Sarcocystis.     

Ultrastructure of sarcocysts from water buffalo in India

The ultrastructure of sarcocysts of macro- and microscopic species of Sarcocystis was compared from naturally infected water buffalo from India. Grossly visible sarcocysts had walls consisting of cauliflower-like villar protrusions, typical of S. fusiformis. The sarcocyst wall of the microscopic species of Sarcocystis was 6.4 microns thick and consisted of tightly packed conical villar protrusions that were 9.6 microns long and 3.7 microns wide at the base. At approximately 3 microns above the base, the distal two-thirds of the villar protrusion became conical shaped and was bent laterally at an angle of 45 degrees to the sarcocyst surface. The granular layer beneath the villar protrusions was 0.9 microns thick. In S. levinei the granular layer was 1.9 microns thick, the villar protrusions were narrow and it had a highly undulating primary cyst wall. Whether the microscopic S. levinei-like sarcocysts of Indian and Malaysian water buffalo are distinct species of Sarcocystis will require further investigation.     

Morphology of Sarcocystis gigantea in experimentally-infected sheep

The development of the parasite and lesions was studied in 32 sheep killed 10 days to 47 months after inoculation with Sarcocystis gigantea sporocysts from cats. At 21-42 days post-inoculation (d.p.i.), there was a mild encephalitis, but organisms were not seen in the brain. Immature sarcocysts were detected from 40-84 d.p.i. The cyst wall was not measurable by light microscopy at 40 d.p.i., but was 1.5-2 microns thick at 84 d.p.i. At 119 d.p.i. both immature cysts containing only metrocytes, and mature cysts containing both metrocytes and merozoites, were present. These mature cysts did not have a secondary cyst wall. A mature cyst, 350 microns in length, was found in a sheep killed at 8 1/2 months p.i. At 10 m.p.i. cysts were up to 0.5 mm long and a secondary cyst wall was present. At 47 m.p.i. cysts were 2-5 X 4.5-7.5 mm, and were found only in the muscles of tongue, oesophagus, pharynx and flank.     

Experimental infection with Sarcocystis medusiformis in sheep

The development of the parasite was studied in 48 sheep killed between 188 and 1132 days after experimental inoculation with Sarcocystis medusiformis sporocysts from cats. Immature sarcocysts were present at 188 days post inoculation (d.p.i.). At 331 d.p.i. macroscopic sarcocysts with an elongate fusiform appearance were seen in the laryngeal, abdominal and diaphragm musculature. The largest cyst measured 2 mm in length by 0.5 mm in width at 331 d.p.i.; histologically they contained metrocytes at the periphery of the cyst with more densely staining merozoites in the central region. By 443 d.p.i. typical 'thin' cysts 2-3.5 mm in length were seen in the flank and external thoracic muscles. By 765 d.p.i. sarcocysts were 5 mm in length. The ultrastructure of the cyst wall of these cysts resembled that of S. medusiformis. At 1132 d.p.i. sarcocysts measured 4 mm X 0.5 mm.     

Ultrastructural differentiation between sarcocysts of Sarcocystis hirsuta and Sarcocystis hominis

The ultrastructure of macroscopic (2-7 mm) Sarcocystis hirsuta sarcocysts from naturally infected cattle from New Zealand was compared with the ultrastructure of 222-day-old S. hominis in experimentally infected cattle in the United States. The villar protrusions of S. hirsuta were approximately 8 microns long, constricted at the base, expanded laterally in the mid-region and tapered distally. Some of the villar tips were folded to form two to four conical projections. The distal portion of the villar protrusions was bent at an angle of 45-90 degrees to the sarcocyst surface. The villar core contained numerous microfilaments and rows of electron-dense granules. The villar protrusions of S. hominis were cylindrical, oriented nearly perpendicularly to the sarcocyst surface, not constricted at their base and contained relatively few electron-dense granules. Although the sarcocysts of S. hirsuta were indistinguishable from those of S. hominis by light microscopy, they were distinguishable ultrastructurally.     

Isolation of Sarcocystis falcatula from the South American opossum (Didelphis albiventris) from Argentina.

Sarcocystis sporocysts from the intestines of four opossums (Didelphis albiventris) from Argentina were identified as Sarcocystis falcatula based on schizogonic stages and pathogenicity to budgerigars (Melopsittacus undulatus). Seven budgerigars fed sporocysts from the opossum feces died of acute sarcocystosis 8, 9, 11, 12, and 14 days after inoculation. Schizonts and merozoites found in the lungs and other organs of the budgerigars were identified as S. falcatula based on structure and immunoreactivity with S. falcatula-specific antibody. Sarcocystis falcatula was also isolated in bovine monocyte cell cultures inoculated with lung tissue from a budgerigar that died nine days after ingesting sporocysts. Two budgerigars inoculated subcutaneously with 1,000,000 culture-derived S. falcatula died 11 and 12 days post-inoculation. This is the first report of S. falcatula infection in South America.     

Coyote as a final host for Sarcocystis species of goats, sheep, cattle, elk, bison, and moose in Montana

Tissues (1 kg) from sheep, goats, cattle, moose, bison, or elk naturally infected with Sarcocystis species were fed to one to four Sarcocystis-free coyotes and the number of sporocysts in feces and intestines were counted. All 12 coyotes fed naturally infected tissues shed Sarcocystis in feces, with a prepatent period of 9 to 15 days. The four coyotes fed infected beef had 15, 25, 113, and 201 million sporocysts in their feces and intestines. The coyotes fed elk, moose, or bison had 2.5, 15, and 2.5 million sporocysts in their intestines, respectively. Sporocysts in feces of coyotes fed musculature of cattle, sheep, goats, and elk were structurally similar to those described previously from the feces of dogs. This is evidently the first report of the completion of life cycle of Sarcocystis species in moose and bison. Cross-transmission experiments indicated that one species of goat Sarcocystis completes its life cycle in both dog and coyote and that the ovine Sarcocystis is not transmissible to goats.     

Sarcocystis infections in Georgia swine

Tissues from 168 mature sows obtained at slaughter in northern and southern Georgia were examined for infection with Sarcocystis spp. Digestion techniques revealed zoites in 28 (16.5%) sows. Infected meat was fed to laboratory-reared dogs, cats, raccoons (Procyon lotor), and opossums (Didelphis marsupialis). Dogs and raccoons shed sporocysts (8.3 mum X 11.2 mum) 12 to 14 days after infection. Cats and opossums were refractory to infection. Sarcocystis-free pigs were infected with 50,000 sporocysts of swine-dog origin. Tissues from laboratory-infected swine were fed to dogs and raccoons. Both species shed sporocysts 14 days later. This is the 1st time in which a definitive host has been demonstrated for species of Sarcocystis occurring in North American swine. The raccoon constitutes a new definitive host for S suicanis Erber 1977. In contrast to previously reported low prevalences of Sarcocystis infections in swine, the relatively high prevalence reported here indicates that S suicanis may be of importance to swine producers in Georgia.