Fine structure and development of Sarcocystis aucheniae in llamas

Isolated cysts of Sarcocystis aucheniae of the llama (Lama glama) were fed to one dog and one cat. Only the dog excreted sporocysts, measuring 13.1-15.7 (15.0 +/- 0.54) X 9.0-11.3 (10.4 +/- 0.36) micron after 11 days for 21 days. A second cat, which had ingested meat of a llama containing macrocysts of S. aucheniae as well as sarcosporidial cysts visible only under a microscope also did not excrete sporocysts. The cysts of S. aucheniae are surrounded by a folded primary cyst wall forming cauliflower-like protrusions into the muscle fibre. The protrusions contain numerous microfilaments. In addition, the primary cyst wall forms numerous tiny vesicles. The parasitized muscle fibre is located in a large cavity within the normal muscle tissue. The cyst wall of S. aucheniae is similarly structured to that of S. gigantea of the sheep.     

Immunoglobulin subclass responses of wild brown rats to Sarcocystis singaporensis

Immunoglobulin subclass responses of wild brown rats (Rattus norvegicus) from southeastern Asia to the endemic cyst-forming coccidian Sarcocystis singaporensis were characterised. The antibody response of brown rats to wild-type parasites (high reproductive capacity) showed a Th1 profile during acute infection, namely elevated concentrations of parasite-specific IgG2b and IgG2c and absence of IgG1. Chronic infection (bradyzoite development) resulted in a mixed Th1/Th2 pattern whereby significant concentrations of IgG1 appeared. A primary infection with 1000 sporocysts eight days before challenge induced protection, accompanied by significant concentrations of IgA and IgG2, particularly IgG2a. Western blot analysis of rat sera, using sporozoite and bradyzoite-extracts as antigen, revealed that IgG1, IgG2a, and IgG2b predominantly recognised molecules between 70-80 kDa in one or the other stage. Some of the antibodies were possibly directed against a 79 kDa heat shock protein of sporozoites. An apparent unresponsiveness to molecules in the low molecular weight range, particularly of bradyzoite antigens, was observed. This was abrogated by infection of rats with an avirulent strain of S. singaporensis (low reproductive capacity) indicating that a parasite that was less adapted to its host provoked a stronger immune response. These results suggest the existence of an immune evasion strategy used by Sarcocystis and show that wild rodents may be suitable as immunological research objects, reflecting the natural situation.     

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 fine structure of the dorsal column nucleus and the nucleus of bischoff of the python (Python reticulatus)

Three types of neuronal perikaryal profiles were identified in the dorsal column nucleus and the nucleus of Bischoff of the python (Python reticulatus). Type I neuronal profiles are large (diameters 12–20 μm) with a deeply indented uncleus. The cisterns of rough endoplasmic reticulum (rER) are mostly randomly dispersed. Axosomatic synapses are few. Type II neuronal profiles (9–11 μm) have a smooth, round, or slightly oval nucleus. Several small stacks of rER are present. Type III neuronal profiles (8–10 μm) have little cytoplasm. The nuclear margin is irregular but not deeply infolded. The rER usually consists of a single long perinuclear ribosome-studded cistern. Two types of astrocytic profiles have been identified. Both types contain abundant filaments. Type I astrocytes are large cells, and the nucleus is very irregular in shape. Type II astrocytes are smaller and are found among the myelinated axons in the dorsal funiculus. Two classes of axon terminals have been identified. One class contains round synaptic vesicles (R profiles) and the other flattened vesicles (F profiles). Some R profiles are small (SR profiles), others are large (LR profiles). Some R profiles also contain a few large, dense-cored vesicles. The R and F profiles establish axodendritic and axoaxonal synapses, some of which are located in the synaptic glomeruli and others in the extraglomerular neuropil. In most of the axoaxonal synapses, the presynaptic element is an F profile and the post synaptic element an LR profile. Occasionally, LR profiles are presynaptic to F profiles. The findings in the python are compared with those of the dorsal column nuclei of the rat, cat, and monkey.     

Fine structure of the oocyst walls of Isospora serini and Isospora canaria and excystation of Isospora serini from the canary, Serinus canarius L.

Oocysts of Isospora serini and Isospora canaria, from the canary Serinus canarius, were broken, added to a cell suspension, fixed in Karnovsky's fluid, and studied in the electron microscope. The oocyst wall of each species had an electron-lucent inner layer, a more osmiophilic middle layer and an outer layer of electron-lucent (I. serini) or electron-dense material interspersed with some electron-lucent material (I. canaria). A few, relatively large lipid-like bodies were present in the outer or middle layer of the oocyst wall of I. canaria. As many as 9 membranes were present in the oocyst wall of I. canaria and 3 in that of I. serini. When exposed to a trypsin-sodium taurocholate fluid, sporozoites of I. serini excysted from 5-month-old sporocysts in vitro, but not from sporocysts stored for more than 6 months. No excystation occurred in 15-month-old I. canaria sporocysts. Similarities and differences in excystation between I. serini and other Isospora, Eimeria, and Sarcocystis species are discussed.     

Reduction of transmission stages concomitant with increased host immune responses to hypervirulent Sarcocystis singaporensis, and natural selection for intermediate virulence.

Parasite virulence (pathogenicity depending on inoculum size) and host immune reactions were examined for the apicomplexan protozoan Sarcocystis singaporensis. This parasite is endemic in southeastern Asia and multiplies as a proliferation (merozoite) and transmission stage (bradyzoite) in rats. Virulence in wild brown rats of parasites freshly isolated in the wild (wild-type) was surprisingly constant within the endemic area and showed an intermediate level. In contrast, serially passaged parasites either became avirulent or virulence increased markedly (hypervirulence). Production of transmission stages was maximal for the wild-type whereas numbers were significantly reduced for hypervirulent and avirulent (shown in a previous study) parasites. Analyses of B and T cell immunity revealed that immune responses of WKY rats to the transmission stage were significantly higher for hypervirulent than for wild-type parasites. These results suggest that it is the immune system of the host that is not only responsible for reduction of transmission stages in individual rats, but also could act as a selective force that maintains intermediate virulence at the population level because reduction of muscle stages challenges transmission of S. singaporensis to the definitive host. Collectively, the presented data support evolutionary theory, which predicts intermediate rates of parasite growth in nature and an ‘arms race’ between host immunity and parasite proliferation.     

Fine structure of the down feather during its early development

The down feather of the chick embryo has been examined by electron microscopy during three distinct stages of its early development; the presumptive stage, represented by dorsal skin of an area from which the feather organ will arise; the thickening stage, during which areas of the basal epidermis form spurs projecting into the mesenchyme, and the latter condenses under a thickened area of the epidermis; the elevation stage, at which time the basal epidermis flattens, the entire epidermis increases in thickness, and the underlying mesenchyme becomes more compact. As development proceeds the rough endoplasmic reticulum of the epidermal cells dilates, but during the elevation stage begins to flatten, and Golgi is observed with increasing frequency. The mitochondria do not appear to differ except for those in the periderm during the presumptive stage, in which case they reveal a vacant matrix and irregular cristae. Evidence is presented for actual contact between basal epidermal spurs and filopodia of cells within the mesenchyme, some of which contain numerous vesicles. The basal epidermal spurs are also seen in intimate association with collagen and anchor filaments and a network of reticulin. Evidence is also presented for the presence of neuronal elements within the mesenchyme during the thickening stage. Cross sections of cell processes within the condensations of the mesenchyme resemble unmyelinated nerve fibers, and cross sections of filopodia similar to arborizing axons abound at and within the basal lamina of both the thickening and elevation stages. Further support for the presence of nerve fibers within the mesenchyme comes from positive staining results with Bodian's and Ungewitter's methods. This comparative study of three stages of early development of the feather organ serves as a basis for more detailed investigations of each stage.     

A pathogenic new species of Eimeria from the pygmy rabbit, Brachylagus idahoensis, in Washington and Oregon, with description of the sporulated oocyst and intestinal endogenous stages

In January 2003, fecal samples from 13 live pygmy rabbits, Brachylagus idahoensis (Merriam, 1891), were collected at the Oregon Zoo in Portland, Oregon, and sent to the University of New Mexico (UNM), Albuquerque, New Mexico, to be examined for coccidia. In July 2004, 14 more fecal samples were collected and sent to UNM, 6 from some of the same rabbits and 8 from 16 other rabbits (4 were pooled samples from siblings). In addition, tissue sections from 3 dead rabbits (2 from the Oregon Zoo, 1 from Washington State University) also were examined. Two of 4 (50%) pooled fecal samples and 8 of 17 (47%) 1-rabbit samples were positive for a single species of Eimeria, which we describe here as a new species. Sporulated oocysts were subspheroidal, 25.6 x 23.8 (22-28 x 21-27) microm, with a length:width (L:W) ratio of 1.1 (1.0-1.2). A micropyle (approximately 2 microm wide) and 0-1 polar granules were present, but an oocyst residuum was absent. Sporocysts were ellipsoidal, 13.4 x 8.1 (11-16.5 x 7.5-9) microm, with a L:W ratio of 1.7 (1.3-2.2), and they had a Stieda body and sporocyst residuum. Tissue sections showed a heavy infection of the villous epithelial cells of the proximal and mid-small intestine with coccidial endogenous stages, but no stages were found in liver hepatocytes. Meronts with approximately 46 (26-70) merozoites per infected cell appeared to be fully developed and were subspheroidal, 14.8 x 13.9 (13-18 x 10.5-16.5) microm. Developing macro- and microgamonts were indistinguishable from each other and were spheroidal to subspheroidal, 10.4 x 9.5 (9-11 x 7.5-10.5) microm. Mature macrogamonts were spheroidal to subspheroidal, 14.2 x 13.7 (12-17 x 11-16) microm, and mature microgamonts were smaller and subspheroidal, 11.9 x 10.8 (10.5-13 x 9-12) microm. This eimerian seems to be extremely pathogenic to young pygmy rabbits, and given the precarious nature of this unique genetic population, it appears to be an emerging pathogen that deserves immediate further study.     

Fine structure and development of phloem sieve tube content

Summary The content of the mature sieve tube ofAcer phloem consists of a network of 80–100 å fibrils. The development of these fibrils from the fibrous masses found in the developing sieve tube is traced and their possible functional significance in translocation discussed.     

Fine structure and development of the silver and golden cuticle in butterfly pupae

Pupae of the butterflies Danaus chrysippus and Helioconius charitonius display characteristic patterns of golden spots, while the pupae of the genera Euploea and Amauris exhibit metallic lustre over most of their surface; E. core and midamus more golden, A. ochlea and niavius more silvery. The absolute reflectance exceeds 80% at wavelengths longer than 550 nm, but drops more or less steeply at shorter wavelengths (shown by microspectrophotometry for E. core and A. ochlea; in all species this effect is caused by constructive interference of the incident light at Multiple Endocuticular Thin Alternating Layers (METAL cuticle). Dense, cuticular D layers alternate with clear, watery C layers and form over 200 double layers. The thickness of the D layers is fairly constant throughout the stack, whereas the C layers systematically increase and decrease in thickness, thus causing the broad bandwidth of the reflector. Connecting filaments, traversing the C layers in zig zag course, probably secure the mechanical stability of the arrangement. After drying, the C layers have vanished and the lustre is lost; the cuticle is now perfectly transparent, except for D. chrysippus, where it is partly transparent and partly yellow. The metallic reflectance develops between 20 and 30 hr after pupal ecdysis, starting with blue colours which change via green to gold or silver. About half a day before emergence of the imago, the reflection fades again via the opposite colour sequence. Coincident with these colour changes, the METAL cuticle is being deposited and decomposed, respectively. The deposition zone immediately above the apical epidermal microvilli consists of about three helicoidal lamellae as in normal, non-reflecting cuticle. The METAL cuticle is formed abruptly at the outer border of the deposition zone, possibly during condensation of the cuticular microfibres. The periodicity it is suggested is controlled either directly by the epidermal cells or indirectly via appropriate self-assembling processes.     

Eimeria tenella: further studies on the development of the oocyst

AIM: The present study investigated the processes of macrogametogenesis and oocyst formation of Eimeria tenella (Xiamen strain), including the formation of wall-forming body1 (WFB1) and wall-forming body 2 (WFB2), the club-shape body and the origin of the residual body during the transformation from a macrogamete to an oocyst. METHOD: Transmission electron microscopy was used to follow ultrastructural changes of the organelles during parasite development. Frozen section techniques and special staining were used to determine the chemical composition of the club-shape body. RESULTS: Electron lighter WFB1 appeared earlier than the electron denser WFB2 during the process of cyst wall formation. WFB2 appeared to play a key role in cyst wall formation, whereas WFB1 may have a limited role in the wall-forming process. When two last generation merozoites entered the same host cell simultaneously, one of them grew well, but the other one was developmentally retarded, and became a residual body. Our study indicates that the content of the club-shape body are lipoidal in nature, not amyolpectin as suggested previously, because they stained black by Sudan black-B. CONCLUSIONS: During of macrogametogenesis and oocyst formation of E. tenella (Xiamen strain), WFB2 plays a major role in cyst wall formation. The residual bodies come from the undeveloped macrogametes. The club-body is lipoid; and lipometabolism is important energy resource in E. tenella development.