A new species of Sarcocystis (Apicomplexa: Sarcocystidae) from the black bear (Ursus americanus)

Infection with Sarcocystis species is common in herbivores but is rare in bears. Histological sections of 374 black bears (Ursus americanus) from Pennsylvania were examined for sarcocysts. In total, 3 sarcocysts were found in 3 bears, with 1 sarcocyst per section. Sarcocysts from 2 bears were considered a new species, Sarcocystis ursusi. Sarcocysts of S. ursusi n. sp. were microscopic and contained only bradyzoites. By light microscopy, the sarcocyst wall was thin (< 0.5 microm thick) and had minute serrations. Ultrastructurally, the serrations on the sarcocyst wall consisted of villar protrusions (Vp) that were mostly 0.5 microm long. The Vp had bundles of electron-dense microtubules that were as wide as long; these microtubules extended deep into the ground substance layer, a feature that distinguished this species from unnamed sarcocysts from black bear. Bradyzoites were 4.8-6.0 microm long. The sarcocyst from the third bear was structurally different from S. ursusi; its sarcocyst wall was approximately 2 microm thick and had finger-like villi on the cyst wall giving the sarcocyst wall a striated appearance.     

Light and electron microscopical study on sarcocysts from muscles of the rhesus monkey (Macaca mulatta), Baboon (Papio cynocephalus) and Tamarin (Saguinus(=Oedipomidas) oedipus).

Sarcocystis cysts from muscles of monkeys (baboon, tamarin and rhesus monkey) were studied by means of light and electron microscopy. The differences in the morphology of the cyst wall and parasites clearly indicate that the three monkey species examined were each parasitized by at least a specific Sarcocystis species being not identical with S. nesbitti or S. kortei. The large numbers of cysts found within the muscle fibres point out the important role that have these monkeys as intermediate hosts in the life cycle of Sarcocystis species, where the final hosts are still unknown.     

Ultrasonic assessment of pregnancy and fetal development in three species of macaque monkeys

Serial ultrasonic assessments of gestational sac (GS) and fetal biparietal diameter (BPD) were performed in the Japanese monkey (Macaca fuscata fuscata), rhesus monkey (Macaca mulatta), and crab-eating monkey (Macaca fascicularis). In the Japanese monkey, GS increased linearly for 3-8 weeks, whereas BPD increased in a linear-quadratic manner over 8 weeks to term. Ultrasonic assessments of spontaneously aborted fetuses with BPD growth retardation and diagnosis of a pelvic chocolate cyst also were reported.     

Sarcocystis speeri N. sp. (Protozoa: Sarcocystidae) from the opossum (Didelphis virginiana)

The North American opossum (Didelphis virginiana) is host to at least 3 species of Sarcocystis: Sarcocystisfalcatula, Sarcocystis neurona, and a recently recognized Sarcocystis sp. A new name, Sarcocystis speeri, is proposed for the third unnamed Sarcocystis. Immunodeficient mice are an experimental intermediate host for S. speeri. Sarcocystis speeri sporocysts are 12-15 x 8-10 microm in size, and its schizonts are found in many organs of mice. Sarcocysts of S. speeri are found in skeletal muscles and they are up to 5 mm long and filiform. By light microscopy, the sarcocyst wall is thin (<1 microm thick); ultrastructurally, the cyst wall is up to 1.8 microm thick and has characteristic steeple-shaped villar protrusions surmounted by a spire. Sarcocystis speeri schizonts are morphologically and antigenically distinct from schizonts of S. neurona, and S. speeri sporocysts were not infective to budgerigars (Melopsittacus undulatus).     

Neuronal ceroid-lipofuscin storage in a cynomolgus monkey (Macaca fascicularis)

The light and electron microscopical characteristics of non-age-related neuronal ceroid-lipofuscinosis in a young cynomolgus monkey (Macaca fascicularis) are described. The pigment in the neuronal cytoplasm was PAS-positive, sudanophilic, weakly acid-fast, and positive with Schmorl's and aldehydefuchsin stains for lipofuscin. Ultra-structurally, it appeared as membrane-bound, electron-dense aggregates within the cytoplasm of neurones of both brain and spinal cord.     

Ultrastructural development of the sarcocyst of Sarcocystis rauschorum (Apicomplexa: Sarcocystidae) in the varying lemming Dicrostonyx richardsoni

The development of the sarcocyst of Sarcocystis rauschorum in its intermediate host was studied. Lemmings were orally administered sporocysts of S. rauschorum obtained from snowy owls (Nyctea scandiaca). Beginning at 9 days postinoculation (DPI) and at various intervals to 84 DPI, skeletal muscle tissue taken from the infected lemmings was examined by electron microscopy. At 9 DPI the sarcocysts contained few metrocytes and the cyst wall was flat. The metrocytes underwent endodyogeny, and within a few days the cyst wall of the rapidly growing sarcocyst developed numerous tubulovesicular invaginations into the electron-dense layer, and the wall had a few irregular infoldings. By 21 DPI, banana-shaped bradyzoites appeared, and by 84 DPI the mature cysts were filled with bradyzoites in groups subdivided by septa and by deep infoldings of the cyst wall. The fine structure of the wall remained simple throughout maturation, with no conspicuous invagination or protrusion. The sarcocyst produced in response to S. rauschorum is unlike those from many species of Sarcocystis, which have complex walls that change markedly as the sarcocysts mature; however, its simple appearance is similar to other species that have rodents as intermediate hosts and raptorial birds as definitive hosts.     

Sarcocystis mephitisi n. sp. (Protozoa: Sarcocystidae), Sarcocystis neurona-like and Toxoplasma-like infections in striped skunks (Mephitis mephitis)

Two structurally distinct types (A, B) of microscopic sarcocysts were found in muscles of 4 of 5 feral skunks. Type A sarcocysts had sarcocyst walls of up to 6 microm thick. The villar protrusions (Vp) on the sarcocyst wall were up to 5 microm long. The Vp were constricted at the base, expanded in the middle, and had a blunt tip. Numerous microtubules were present in the Vp and in the granular layer. Bradyzoites were up to 11 microm long and up to 3.2 microm wide. Based on the distinctiveness of the Vp, a new name, Sarcocystis mephitisi is proposed for type A sarcocysts. Type B sarcocysts had a relatively thin (approximately 1-2 microm thick) sarcocyst wall and the Vp were slender and tapered toward the tip. These sarcocysts were structurally similar to S. neurona sarcocysts. A Toxoplasma gondii-like tissue cyst was found in a section of tongue of 1 of the 4 skunks.     

An electron microscopic study of the macro- and microcysts of coccidia in the genus Sarcocystis from buffalo

Two sarcosporidian species from muscles of the water buffalo examined in Azerbaijan were studied. The one making macrocysts was identified as S. fusiformis, whereas the other producing microcysts appeared to be not S. levinei as it might have been expected. Indeed, the microcysts found in the muscles of the buffalo in the slaughterhouse [correction of slaughtery] of Baku differed essentially in morphology of the cyst wall from that of S. levinei, sooner resembling that of S. cruzi from cattle. The cyst wall of these microcysts was seen to make deep invaginations towards the ground substance of cysts. From these invaginations some curved channels were grown outlined with a filamentous material. The described pattern of cyst wall is not characteristic of the S. cruzi sarcocyst. No specification was achieved, and the sarcocysts found in the bubaline muscles are designated as Sarcocystis sp. from the buffalo.     

Effects of frozen storage on the structure of sarcocysts in pig muscle and implications in taxonomic studies

The morphology of the cyst wall of Sarcocystis has unique characteristics that can be used in species identification. To find a suitable way to preserve Sarcocystis cyst samples for species identification, by light microscopy and electron microscopy, we recorded the morphological changes in the cysts of Sarcocystis suihominis and Sarcocystis miescheriana from pig muscle, induced by storage at -20 degrees C. Comparisons were made between fresh cysts and those subjected to frozen storage for periods of 3 days, 20 days and 30 days. Results: cyst wall of the two Sarcocystis species appeared unaffected by storage. There was no obvious change in the length, nor in the width of the protrusions after storage (P>0.05), but the structure of the bradyzoite in the sarcocyst was in many cases disintegrated at -20 degrees C in 20 days for S. miescheriana and 30 days for S. suihominis. To our knowledge this is the first report that Sarcocystis cyst in muscle can be stored at -20 degrees C before and remain suitable for ultrastructural morphological study. Consequently, this paper proposes freezing as a convenient storage method for samples used in taxonomic studies of Sarcocystis.     

Redescription of the sarcocysts of Sarcocystis rileyi (Apicomplexa: Sarcocystidae)

The intermediate hosts for Sarcocystis rileyi (Stiles 1893) Minchin 1913 are ducks (Anas spp.), and the striped skunk (Mephitis mephitis) is its definitive host. The structure of sarcocysts from an experimentally infected shoveler duck (Anas cylpeata) fed sporocysts from an experimentally-infected M. mephitis was studied and compared with type specimens from a naturally infected duck. The experimentally infected duck was killed 154 d after feeding sporocysts. By light microscopy the sarcocyst wall was 3-5 microm thick with indistinct villar protrusions. Ultrastructurally, the sarcocyst wall was a type-23 cyst wall with anastomosing villar protrusions that were up to 7.5 microm long. The villar projections contained filamentous structures. The bradyzoites were 12-14 microm long. Structurally, the sarcocyst from the naturally infected and experimentally infected ducks appeared similar.     

Some monoclonal anti-human lymphocyte and class II MHC antigen antibodies do not stain snap frozen Macaca fascicularis lymphocytes or tissue sections.

Experiments were designed to stain snap frozen and fixed Macaca fascicularis peripheral blood mononuclear cell (PBMC) preparations and colon sections assuming that monoclonal mouse anti-human lymphocyte and class II major histocompatibility complex antigen antibodies would cross-react with the monkey counterparts to the human antigens. Most of the monoclonals used in this study did not stain the monkey frozen and fixed tissues in immunoenzymatic protocols despite adequate staining of control human tissues. The fact that snap frozen Macaca fascicularis PBMCs and tissue sections were not always stained is of practical importance for experimental design in Macaca fascicularis using anti-human reagents.     

Ultrastructural studies of Sarcocystis cruzi (Hasselmann, 1926) Wenyon, 1926 infection in cattle (Bos taurus): Philippine cases

This paper documents the first report of Sarcocysti s cruzi infection in domesticated cattle (Bos taurus) in the Philippines. Fusiform-shaped microscopic sarcocysts (183-578 microns long and 20-98 microns wide) with distinct septae were found in the skeletal, striated and heart muscle. The sarcocyst wall or parasitophorous vacuolar membrane, 1.37-2.75 microns thick consisted of closely-packed villar protrusions 80-400 nm in dm. Middle and distal segments of VP were bent approximately 90 degrees parallel to the cyst wall surface. The villar core lacked microtubules, and at some points, the distal ends of the VP collectively formed conical tufts. Primary cyst wall had numerous 70-100 nm bubble-like undulations, and the ground substance was 0.25-0.5 micron in thickness. The ultrastructure of S. cruzi cyst wall typifies the Type 7 sarcocyst wall, and bears close similarities with the Philippine and the Vietnam strain of bubaline Sarcocystis levinei.     

Sarcocystis dubeyella n. sp. and Sarcocystis phacochoeri n. sp. (Protozoa: Sarcocystidae) from the Warthog (Phacochoerus aethiopicus) in South Africa

ABSTRACT Sarcocystis dubeyella n. sp. and S. phacochoeri n. sp. from muscle fibers of the skeletal musculature of two warthogs in South Africa are described by light and and electron microscopy. Sarcocystis dubeyella sarcocysts are macroscopic (up to 12 mm long and 1 mm wide), with a parasite-induced encapsulation of the host muscle fiber in which the plasma membrane of the latter remained unaltered. The sarcocyst wall is characterized by evenly arranged, irregularly semicircular or rectangular villar protrusions (5.0 T. 2.8-11.0 μm) with indented margins and no specific content. Sarcocystis phacochoeri formed filiform microcysts (up to 4 mm long and 0.13 mm wide). Its cyst wall is provided with tightly packed, molarlike villar protrusions (1.6-3.3 T. 1.7-3.3 μm), with smooth margins, hollow on one side, and with longitudinal condensations of the fine granular matrix at various locations in the interior.     

Blockage of urine by intravesical ejaculate in cynomolgus monkeys

BACKGROUND: The present communication reports intravesical semen coagulation and formation of a larger precipitate in two Cynomolgus monkeys. METHODS: Ultrasound of the urinary bladder and light microscopy of intravesical coagulates. RESULTS: These monkeys suffered from complete blockage of urine output and surgery was required to remove the sperm mass. Microscopic examination of the urine revealed millions of sperm as a cause of the mass and the blockage of urine output. CONCLUSIONS: Retrograde ejaculation of sperm may cause coagulation of ejaculates in the bladder of the cynomolgus monkey Macaca fascicularis. However, involvement of sperm mass in blockage of urine passage has not been described in this species.     

Radiation-induced translocations in spermatogonial stem cells of Macaca fascicularis and Macaca mulatta

3 adult monkeys, one Macaca fascicularis and two Macaca mulatta, were whole-body irradiated with 1 Gy gamma-rays (60 Co). Reciprocal translocations induced in spermatogonial stem cells were scored as translocation multivalents in primary spermatocytes from 7.5 to 27.5 months after exposure. The translocation yields ranged from 4.1% at the earliest to 1.8% at the latest sampling interval. No significant differences were observed in the responses of the individual animals. A decline in the translocation frequencies with time after treatment was found in all 3 animals. The present data are different from those reported for testicular X-irradiation of the rhesus monkey Macaca mulatta (van Buul, 1980; Lyon et al., 1976) in that the translocation yields are higher. They are consistent with the results reported for testicular gamma-irradiation of the crab-eating monkey Macaca fascicularis (Matsuda et al., 1984, 1985). In view of the present results it appears unlikely that a species difference exists within the genus Macaca in the sensitivity of spermatogonial stem cells to the induction of translocations by ionizing radiation.     

Sarcocystis nesbitti n. sp. from the Rhesus Monkey

SYNOPSIS. Sarcocystis nesbitti n. sp. was found in the skeletal muscles of 2 of 44 Macaca mulatta monkeys. The cysts were 100-1100 by 165 μ with a wall 1.2-1.6 μ wide and contained trophozoites 7–9 by 2–3 μ. Sarcocystis kortei was found in 1 of the rhesus monkeys, 1 Erythrocebus patas and 1 Cercopithecus mitis. Its cysts were 400-800 by 100-150 μ, with a wall 5-6 μ thick and trophozoites 9-14 by 3-4 μ.     

Structural and functional characterization of cyst cells in Sarcocystis sp. (Sporozoa, Apicomplexa)

By means of light and electron microscopy, the structural pattern of muscle cysts (sarcocysts) was examined for the four species of the genus Sarcocystis: S. muris (from murine skeletal muscles), Sarcocystis sp. and S. fusiformis (from, respectively, heart and skeletal muscles of buffalo), and S. ovifelis (from ovine tong muscles). The orderly fashion of the interior of the cyst is attained by partitition of its space into numerous compartments with the involvement of the intermediate filaments. These, in their turn, are bound to each other by thin filaments to make eventually a common filamentous net. The net limits separate groups of cells referred to as cyst zoites. The common net of filaments and microtubules (when present) may be regarded not only as the organizer of the cyst interior cytoskeleton, but also as the main mechanism of substance transportation in various directions: from the host cell to the sarcocyst, and within or outside the cyst. The role of dedifferentiation, proliferation and differentiation processes is suggested in the establishment of the fixed sequence of events throughout the unidirectional development of cyst cells and their interaction, from precystic meronts to cyst merozoites (gamonts). Special attention is paid to metrocyte morphogenesis and functioning. In the present work, metrocytes subjected to apoptosis were recognized. It is suggested that phenomenon of programmed cell death in metrocytes may be associated with the control of cell number in mature and ageing sarcocysts.     

Surgical excision of an ameloblastic odontoma in a cynomolgus monkey (Macaca fascicularis)

A surgical approach used for removal of maxillary tumors in humans was modified to excise a maxillary tumor in a cynomolgus monkey (Macaca fascicularis). This is an effective technique for excising tumors, recovering fractured canine teeth, or other health problems of the maxillary region in Macaca sp. The described approach limits facial disfigurement and enhances the probability of return to required function for future research utilization. This approach minimizes the extensive degree of post-operative intensive care.     

Proteolytic enzymes from recombinant Streptomyces lividans TK24

Different proteases from the culture fluids of recombinant Streptomyces lividans strains were isolated. Several individual proteases were separated and characterized. A chymotrypsin-chylike activity (CLA) was identified that specifically degrades a fusion protein between the alpha-amylase inhibitor from S. tendae (Tendamistat, HOE467) and proinsulin from the monkey Macaca fascicularis. The effective chemical inhibition of the degrading enzyme is demonstrated.