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.     

Sarcocystis sinensis is an ultrastructurally distinct parasite of water buffalo that can cause foodborne illness but cannot complete its life-cycle in human beings

In this study, we compared the morphology of Sarcocystis sinensis and Sarcocystis hominis, and assessed the infectiousness of S. sinensis for human volunteers. The cysts of S. sinensis were from water buffalo (Bubalus bubalis) and those of S. hominis were from cattle (Bos taurus). Transmission electron microscopy of S. sinensis cysts revealed that the cyst wall had leaning, finger-like protrusions measuring 1.44-5.08 μm in length and without invaginations on the tip surface of the protrusions. In contrast, the cyst wall of S. hominis had upright, finger-like protrusions measuring 9.43 μm×2.42 μm and with vesicle-like invaginations on the tip surface of the protrusions. Scanning electron microscopy revealed that surface of the protrusions was arranged as rectangles in S. sinensis, as compared to tongue-shaped in S. hominis. Other distinguishing features of S. sinensis include a thin ground substrate (GS) zone with microtubules and small, circle-like structures located at the base of the protrusions. Human volunteers, after consuming S. sinensis cysts, produced no sporocysts or oocysts in feces, suggesting that humans could not serve as definitive hosts for S. sinensis. By contrast, many sporocysts and oocysts were passed in feces of a human volunteer 11-29 days after ingestion of S. hominis cysts. These results showed that S. sinensis and S. hominis are separate species and S. sinensis cannot use human being as the definitive host.     

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.     

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.     

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.     

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).     

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.     

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.     

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.     

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.     

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.     

CRANIAL MEDIASTINAL CYSTS IN NINE CATS

Nine cats, from 11 to 17 years of age (mean 13.6 years of age), were diagnosed with a cranial mediastinal cyst. Thoracic radiographs in all cats were characterized by an increased soft tissue opacity in the cranial mediastinum confirmed to be a cyst by ultrasonography or necropsy. Ultrasonographically cysts appeared as an anechoic mass. A low-cellularity clear fluid was obtained on aspiration. The majority of the cats (n = 8) presented for unrelated conditions with no signs of respiratory distress. No treatment for the cyst was pursued except for drainage during ultrasonographic-guided aspiration in several cats. On follow-up of eight cats, none were symptomatic for the cyst from 3-45 months after diagnosis. Mediastinal cyst should be considered when a cranial mediastinal mass is evident radiographically in an older cat. The majority of feline cranial mediastinal cysts are benign with no need for treatment.     

Association of eosinophilic myositis with an unusual species of Sarcocystis in a beef cow.

The carcass of a mature cow had numerous, disseminated lesions typical of eosinophilic myositis. To elucidate the nature and possible cause of the lesions, histological sections were examined by light microscopy and selected areas were removed and processed for electron microscopy. The lesions were granulomatous in nature. Each granuloma contained at its centre an intact or ruptured sarcocyst associated with degenerate muscle fibers. Surrounding this was a layer of epithelioid cells and an intense accumulation of inflammatory cells, most of which were eosinophils. The primary cyst wall of the sarcocysts in these granulomas consisted of hair-like protrusions that featured many unusual electron-dense bodies. Sarcocysts with ultrastructures characteristic of Sarcocystis cruzi and Sarcocystis hirsuta were also present in muscle from the same animal, but these sarcocysts lacked any associated cellular responses. The eosinophilic myositis in this case appeared to be associated with sarcocystosis of an unknown species. Possibly, the inflammatory reaction was due to the host-parasite interaction in an unusual host.     

Experimental inoculation of domestic cats (Felis domesticus) with Sarcocystis neurona or S. neurona-like merozoites

Sarcocystis neurona is the parasite most commonly associated with equine protozoal myeloencephalitis (EPM). Recently, cats (Felis domesticus) have been demonstrated to be an experimental intermediate host in the life cycle of S. neurona. This study was performed to determine if cats experimentally inoculated with culture-derived S. neurona merozoites develop tissue sarcocysts infectious to opossums (Didelphis virginiana), the definitive host of S. neurona. Four cats were inoculated with S. neurona or S. neurona-like merozoites and all developed antibodies reacting to S. neurona merozoite antigens, but tissue sarcocysts were detected in only two cats. Muscle tissues from the experimentally inoculated cats with and without detectable sarcocysts were fed to laboratory-reared opossums. Sporocysts were detected in gastrointestinal (GI) scrapings of one opossum fed experimentally infected feline tissues. The study results suggest that cats can develop tissue cysts following inoculation with culture-derived Sarcocystis sp. merozoites in which the particular isolate was originally derived from a naturally infected cat with tissue sarcocysts. This is in contrast to cats which did not develop tissue cysts when inoculated with S. neurona merozoites originally derived from a horse with EPM. These results indicate present biological differences between the culture-derived merozoites of two Sarcocystis isolates, Sn-UCD 1 and Sn-Mucat 2.     

交叉感染后黄牛与水牛枯氏住肉孢子虫包囊超微结构的比较研究

选用4头7日龄奶牛和4头4～5月龄水牛,用水牛源孢子囊感染黄牛及黄牛源孢子囊感染水牛,同时设感染对照和不感染对照,对交叉感染后黄牛与水牛体内包囊的超微结构进行了比较研究,结果发现两者无结构区别,所有包囊的超微结构均与前人对黄牛和水牛枯氏住肉孢子虫包囊的描述一致,证实水牛与黄牛同是枯氏住肉孢子虫的中间宿主。作者还首次在枯民住肉孢子虫包囊的母细胞和缓殖子发现晶状体。     

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.     

Disseminated sarcocystosis in a cat with lymphosarcoma

Cysts of the protozoan Sarcocystis sp were found in skeletal and cardiac musculature in a 1.5-year-old cat with lymphosarcoma. The cat was FeLV-positive and had grossly visible neoplastic involvement of the spinal cord, mediastinum, bone marrow, and kidneys. Ultrastructural examination of the parasitic cyst wall suggested that the species in this case was different from that described in the only other reported case. It was hypothesized that immunosuppression from FeLV infection permitted an aberrant life cycle with encystment of Sarcocystis sp in this cat.     

Naturally occurring Sarcocystis neurona-like infection in a dog with myositis

Tissue stages similar to those of Sarcocystis neurona, the causative agent of equine protozoal myeloencephalitis, were identified in skeletal muscles of a dog. The dog, a 6-year-old Labrador retriever, was seropositive for Toxoplasma gondii infection and euthanized due to a history of polymyositis and progressive muscular atrophy. Histologically, 30, variably sized, microscopic, intracellular sarcocysts were observed in 60 sections of skeletal muscles taken from the neck, fore limbs and hind limbs. The cysts were only observed in inflamed skeletal muscles, but were mostly in myocytes at the periphery of areas infiltrated with leukocytes. Ultrastructurally, the cyst wall had villar protrusions consistent with sarcocysts. Immunohistochemistry with monoclonal S. neurona antibodies demonstrated positive labeling of zoites in merozoites or schizonts in the skeletal muscle interstitium, but no labeling of the sarcocysts. Initial PCR analysis with primers amplifying a genetic sequence encoding Apicomplexan 18s rRNA, and subsequent PCR analysis with differentiating primers indicated that the genetic sequences had 100% identity with sequences reported for S. neurona.