Morphology of Diplomonad Flagellates: Spironucleus torosa N. Sp. from Atlantic Cod Gadus morhua L., and Haddock Melanogrammus aeglefinus (L.) and Hexamita salmonis Moore from Brook Trout Salvelinus fontinalis (Mitchill)

ABSTRACT. A diplomonad flagellate, Spironucleus torosa n. sp. is described from Atlantic Cod Gadus morhua and haddock Melanogrammus aeglefinus . This is believed to be the 1st confirmed report of Spironucleus from a marine fish. Organisms swimming in the rectal lumen were broadly pyriform to elongate, and measured 10.5–18.6 μm long and 3.2–13.3 μm wide; other elongate organisms were attached to the rectal epithelium, via apical extensions appearing continuous with the microvilli. The posterior end of the body was extended into a caudal projection, on either side of which was a posteriolateral ring-shaped protrusion or torus, with a recurrent flagellum emerging from its centre. A symmetrical system of microtubules and lamellae, forming a "V" in protargol impregnated specimens, supported the flanges of the body surrounding the tori, the tori themselves and the caudal projection. Supranuclear microtubules were an inverted V to U shape in transverse section, and an electron dense band accompanied the cytostomes. Lightly staining homogenous cytoplasm was usually present in the anterior part of the body, the remainder being highly vacuolated with numerous dark granules. In swimming organisms, rough endoplasmic reticulum (RER) was present around the nuclei and cytostomes, and bacteria were occasionally seen in the cytoplasm. In "attached" organisms, RER was reduced, and bacteria were absent. Hexamita salmonis Moore from Salvelinus fontinalis was studied by light and scanning electron microscopy for comparison; its cytoplasm was not highly vacuolated. The two recurrent flagella emerged close together from the blunt posterior end of the body.     

Hexamastix dobelli n.sp., a New Trichomonad Parasitic in the Starred Tortoise

Hexamastix dobelli n. sp. (Trichomonadidae) is described from the large intestine of the starred tortoise, Testu-do elegans Schoepff. This is the first species of Hexamastix to be reported from a chelonian. The organism is pyriform, oval, or round in shape. There are five anterior flagella, of which two are long and three short and a single recurrent flagellum, which equals the short anterior flagella in length. The axostyle is very slender and projects for some distance outside the body. The nucleus contains three to six granules and has no endosome. The pelta is crescent-shaped and the parabasal body is rod-like. Perinuclear granules are present in the anterior half of the body. The parasite is 6.0 to 16.5 μ long, 2.0 to 10.3 μ wide and the diameter of nucleus is 2.5 μ.     

Ultrastructure of In Vivo-Produced Caryocysts Containing the Coccidian Caryospora bigenetica (Apicomplexa: Eimerüdae)

A cotton rat was inoculated orally with oocysts of Caryospora bigenetica from the feces of a rattlesnake. Sixteen days later the rat was euthanized, and portions of the scrotum, foot pad and muzzle were processed for histological sections and transmission electron microscopy. Sporozoites within caryocysts had typical coccidian features such as an anterior and posterior refractile body, centrally located nucleus, micronemes, rhoptries, a conoid, a micropore near the anterior refractile body, a posterior pore, amylopectin granules, lipid bodies, a Golgi-like body, a mitochondrion and subpellicular microtubules. The infected host cell was spherical and surrounded by a fibrous wall-like covering, 0.35–1.00 μm thick. This outer covering, when viewed in stained histological sections, was periodic acid-Schiff (PAS)-positive.     

The Structure of the Oocyst and the Excystation Process of Isospora marquardti sp. n. from the Colorado pika, Ochotona princeps

SYNOPSIS. Oocysts of Isospora marquardti sp. n. from the Colorado pika, Ochotona princeps , have spheroid oocysts, 30.5 (23–36) μm in diameter, and ovoid sporocysts, measuring 19.3 × 12.0 (17–22 × 10–14) μm. A polar body, 2 × 4 μm, a spheroid sporocyst residuum (8.3 μm in diameter), a Stieda body, and a distinct substiedal body (3 × 3 μm) are present. A micropyle and an oocyst residuum are absent. Excysted sporozoites, averaging 3.0 × 18.5 (2–4 × 15–20) μm, contain 2 refractile globules, 1 on each side of the nucleus with a prominent nucleolus.
The sporozoite excystation process using a trypsin-sodium taurocholate fluid is described.     

Ultrastructure of Tachyzoites, Bradyzoites and Tissue Cysts of Neospora caninum

. Dividing tachyzoites of Neospora caninum were 4x3 μm and had ultrastructural characteristics typical for the cyst-forming coccidia. Unusual ultrastructural characteristics of fully-formed tachyzoites included no micropores, 8–12 anterior and 4–6 posterior rhoptries, and a few posterior micronemes. Most tachyzoites were located free in the host cell cytoplasm; only a few occurred within a parasitophorous vacuole. Parasite multiplication appeared to be rapid because most organisms were in various stages of endodyogeny. Neural tissue cysts of N. caninum were 24.3 × 19.2 μm and contained 50–200 bradyzoites (7.3 × 1.5 μm), which lacked micropores. The cyst wall was 0.74–1.12 μm thick and consisted of the primary cyst wall (the parasitophorous vacuole membrane) and a thick granular layer with electron-dense vesicles.     

Endogenous Stages of the Life Cycle of Isospora rivolta in the Dog

SYNOPSIS. Coccidia-free beagle puppies were experimentally infected with a cloned culture of Isospora rivolta oocysts. The endogenous stages were found in the posterior 1/2 of the small intestine, and rarely in the cecum and colon. Maximum numbers of all stages occurred just anterior to the ileocecal valve. Endogenous stages were found in the distal third of the villi, predominantly parasitizing subepithelial cells of the lamina propria; however, stages were occasionally present in epithelial cells. The number of asexual generations could not be determined from their structure, but evidence based on oocyst production suggested that there were at least 2 asexual generations. The schizonts were 17–24 by 12–25 μ and contained 4–24 merozoites, the most common number being 4 or 8. Schizonts with mature merozoites were found as early as 72 hr, but were present in maximum numbers at 96 hr. Merozoites had slender curved bodies and were 10.5–13.4 by 2.3–3.0 μ. Mature gamonts were found by 144 hr. Mature microgametocytes were 13.4 by 8.7 μ and contained 50–70 microgametes. Microgametes had slightly curved tapering bodies (5.8–6.4 by 0.6 μ) with 2 posteriorly directed flagella 11–14 μ long. Mature macrogametes had reticular cytoplasm and a uniformly large nucleus and nucleolus.
The prepatent period was 142–146 hr. The patent period was 13–23 days with an average of 19 days.     

Ultrastructure of Sporozoites and Zoites of Hammondia heydorni

The ultrastructure of sporozoites and zoites of Hammondia heydorni was studied in cultured bovine cells. In addition to ultrastructural features typical of coccidian parasites, H. heydorni sporozoites and zoites contain rhoptries that are located posteriorly as well as anteriorly. Also, sporozoites contain a posteriorly located crystalloid body (1.2 μm in diameter); a small crystalloid body (0.5 μm in diameter) was occasionally seen in the anterior end. Zoites resulting from the 1st division of endodyogeny contain a posteriorly located crystalloid body, which is absent in zoites formed by subsequent divisions. Zoites contain posteriorly located amylopectin granules and a relatively large anterior vacuole which is not present in sporozoites. During penetration, the host cell plasmalemma ballooned laterally around the sporozoite creating a large cavity, which later disappeared. Sporozoites and zoites undergoing cell penetration usually exhibit partially empty anterior rhoptries; no changes occur in posterior rhoptries. Lysosomes fuse with the par-asitophorous vacuole surrounding killed sporozoites but not live sporozoites.     

Coccidian Parasites (Apicomplexa: Eimeriidae) of Nerodia spp. (Serpentes: Colubridae), with a Description of a New Species of Eimeria

Eimeria conanli n. sp. (Apicomplexa: Eimeriidae) is described from intestinal contents and feces of Nerodia erythrogaster transversa and N harteri harteri from northcentral Texas. Oocysts of the new species are ellipsoid in shape. 17.9 × 13.0(15–21 × 12–15) μm, with a smooth, thin, single-layered wall; shape index 1.4 (1.2–1.5). One to several (usually 2) polar granule(s) and an oocyst residuum are present, but a micropyie is absent. Sporocysts are elongate, 12.9 × 5.2 (13–15 × 5–6) -m, apparently without a true Stieda body structure. Each sporoeyst contains an ellipsoid residuum, 3.9 × 3.2 (3–6 × 2–4) μm, and elongate sporozoites, 11.4 × 2.5 (10–14 × 2–3) μm in situ, each with a spherical or subspherical anterior refractile body and spherical to ellipsoid posterior refractile body. In addition to the new species, oocysts of 4 previously described eimerians from colubrid snakes were found in these hosts.     

Spironucleus vortens N. Sp. from the Freshwater Angelfish Pterophyllum scalare: Morphology and Culture

ABSTRACT. A new diplomonad flagellate, Spironucleus vortens n. sp., is described from the intestinal lumen of the freshwater angelfish, ( Pterophyllum scalare ), bred in Florida. Live organisms are pyriform, and measure 12.5–20.5 μm long by 5.0–11.2 üm wide. Scanning electron microscopy shows that the trophozoite bears two compound lateral longitudinal ridges, each originating posterior to three emerging anterior flagella, and continuing posteriorly to the emergence of the posterior flagellum. Each ridge comprises a broad central part, surrounded by a peripheral ridge. At the opening of the flagellar pocket, the broader right peripheral ridge crosses to the other side of the body, and then back again. The posterior end of the body bears two papillae. Transmission electron microscopy shows that the compound lateral ridges are supported by microtubules, and bear microfibrillar structures in discrete longitudinal plaques. The serendipitous growth of S. vortens in a culture system with lip tumor tissue, facilitated axenic cultivation in a modified TYM medium (trypticase, yeast extract, maltose). The flagellate is now routinely maintained in an axenic TYI-S-33 medium (trypticase, yeast extract, iron serum), and is stabilized in the cryopreserved state. Spironucleus vortens is an aerotolerant anaerobe that can be cultured at 25° C, 28° C and 30° C.     

The ultrastructure of zoospores ofAphanomyces euteiches and of their encystment and subsequent germination

Summary The ultrastructure ofAphanomyces euteiches during the periods of zoospore motility, encystment, and germination has been studied. The motile spore has two heterokont flagella inserted laterally into the groove of the zoospore body where each is attached to a kinetosome. The kinetosomes and flagella are anchored into the zoospore body by rootlets comprised of two rows of microtubules with up to 12 microtubules in the outer row and are attached by fine threads to a striate fiber bundle. Secondary microtubules are attached at right angles at regular intervals along the rootlets. An unidentified body, 1.25m in diameter, containing helical fibers 16 nm in diameter is present in each zoospore. This body is situated near the two kinetosomes on the side of the pyriform nucleus opposite the contractile vacuole. The Golgi complex is between the nucleus and the contractile vacuole. The latter is surrounded by a 0.5–1.0m wide zone of Golgi proliferated vesicles. Ribosomes are generally absent from this region. Endoplasmic reticulum containing tubules within the expanded cisternae are also present. Vesicles with striated electron opaque inclusions and vesicles containing a granular cortex and center that developed in previous stages of zoosporogenesis were also present. During encystment of the zoospore the latter vesicles disappear. The two flagella are shed at this time leaving a membrane-bounded granular knob protruding from each of the kinetosome terminal plates. The contractile vacuole becomes disorganized and the zoospore assumes a spherical shape. Cyst wall deposition begins immediately and is completed in 30 minutes. The spore begins to germinate 1 hour following initiation of encystment with the appearance of a bulge in the cyst wall which elongates into a germ tube. Mitotic nuclear division follows.Research supported by the College of Agricultural and Life Sciences Station Project No. 1281.Research assistant and Professor. The advice and assistance of G. A. deZoeten, G. R.Gaard, and S.Vicen are most gratefully acknowledged.     

Two New Menosporine Gregarines, Hoplorhynchus acanthatholius N. Sp. and Steganorhynchus dunwoodyi N. G., N. Sp. (Apicomplexa: Eugregarinorida: Actinocephalidae) from Coenagrionid Damselflies (Odonata: Zygoptera)

ABSTRACT. Hoplorhynchus acanthatholius , n. sp. is described from Enallagma civile , the Civil Bluet damselfly. Trophozoites are solitary, lie in the mesenteron between the peritrophic membrane and the epithelium, and attain a maximum length of 850 μm. Epimerite ovoid to broadly ovoid; anterior margin bearing eight equidistant retroarcuate hooks; attached to protomerite by means of a vermicular stalk. Protomerite ovoid; deutomerite narrowly obvoid. Gametocysts spherical; diam 300 μm, sporulating by simple dehiscence in 48–72 h. Oocysts are characteristic of Menosporinae: smooth, biconical, crcscentic, uniform in size and shape. Steganorhynchus dunwoodyi , n. g., n. sp. is described from the damselfly Ischnura verticalis. The genus is characterized by an epimerite comprising an ovoid papilla enclosed in a retractable, globular sheath, borne on a long vermicular stalk. Trophozoites are solitary, lie in the mesenteron between the peritrophic membrane and epithelium, and attain a maximum length of 605 μm. Protomerite very broadly ovoid; deutomerite obvoid. Gametocysts spherical; diam 258 μm, sporulating by simple dehiscence in 48–72 h. Oocysts are characteristic of Menosporinae: smooth, biconical, crescentic, uniform in size and shape. The population dynamics of H. acanthatholius and S. dunwoodyi among damselfly populations in five Nebraska localities are presented.     

Myxosoma microspora n. sp. (Myxosporidia: Protozoa) parasitic in the gills of Mugil cephalus L.

A new histozoic species of myxosporidian, Myxosoma microspora n.sp., infecting the gill filaments of Mugil cephalus is described. Cysts measuring 0.5–1.0 mm in diameter were found attached to the gill filaments. Spherical or slightly oval, spores 4.8–5.2 μm in diameter, were present and possessed a thin outer mucous envelope which appeared as small conical protuberances at the ends of the equatorial axis. Polar capsules were pyriform in shape, equal in size and measured 1.6–2.0 × 1.0–1.2 μm; the polar filaments were 22–28 μm in length. There was a bean-shaped sporoplasm measuring 3.5 × 1.5 μm. No iodine vacuole was detected when the parasite was stained with Lugol's iodine.     

Meiosis of Primary Spermatocytes and Early Spermiogenesis in the Resultant Spermatids in Newt, Cynops pyrrhogaster in vitro

Dissociated spermatogenic cells were cultivated within the collagen matrix at low cell density. The largest cell type in the culture was identified as the primary spermatocytes by their size and the morphological characteristics revealed by ultra-thin sections. Chromosome analysis showed that about 90% of the cells examined were either in first or second meiosis. Within the collagen matrix, the fates of 282 single primary spermatocytes at meiotic stage in diakinesis or metaphase were followed. In a few days, most of them gave rise to four spermatids, passing through first and second meiotic divisions. About 80% of the spermatids formed motile flagella. They grew about 20–60 μm a day. The final state of the differentiation attained in our culture conditions was the spermatids with localized spherical nuclei and motile flagella, about 500 μm in length after 1-month's culture. Ultra-thin sections of the spermatids show that the rings, neck-pieces, and acrosomes developed in the cells.     

Fine structure of the respiratory organs of the Climbing perch, Anabas testudineus (Pisces: Anabantidae)

An electron microscopic study has been made of the three respiratory organs of climbing perch. The gill structure is similar to that of the other telcosts but the thickness of the water/blood barrier is much greater, being as great as 20 μm in some specimens. The increased thickness is due to a multilayered epithelium which is thinner (3.5–7 μm) over the marginal channel of the secondary lamellae. The other two main layers, basement membrane and pillar cell flange, are relatively thin (about 1 μm).
The pillar cells have a typical structure, but in certain regions they are contiguous with one another and line well-defined blood channels. Some of the columns of basement membrane material in such regions may be common to adjacent pillar cells.
The air-breathing organs are (a) the lining of the suprabranchial chambers , and (b) the labyrinthine plates attached to the dorsal region of branchial arches. Electron microscopy showed that their structure is well adapted for gas exchange, the air/blood barriers being only 0.12–0.3 μm, comprising an epithelial layer, basement membrane, and thin capillary endothelium. The many parallel blood channels of the respiratory islets of both organs are separated by pillar-like structures which differ from the pillar cells of the secondary lamellae. Thus the hypothesis that the air-breathing organs represent modified gills is not supported by this study.
The fine structure of the non-respiratory region of the air-breathing organs is similar to that of the skin, and includes chemoreceptor-like cells. Evidence concerning the possible homology of pillar cells with plain muscle cells is discussed.     

Morphological Comparison of Diophrys scutum (Dujardin, 1841) and Diophrys peloetes n. sp. (Hypotrichida, Ciliophora)

SYNOPSIS. Diophrys scutum , collected from four locations on the New Hampshire coast, ranged from 89–195 7mu; in length, 50–105 μ in width, and 68–88 μ in buccal cavity length. The end of the adoral zone of membranelles (AZM) extends 37–59 μ (average = 45.6 μ) posteriorly in a groove on the right side of the body. Dorsally are five rows of stiff cilia. The silverline system (Chatton-Llvoff technique) appears as a fine meshwork, entirely different from that found in Euplotes or Uronychia . There are two elongate macronuclei (Feulgen reaction) and several micronuclei. Diophrys peloetes n. sp., collected from one location in Alligator Harbor. Florida, ranged from 95–134 μ in length, 62–84 μ in width. and 60–80 μ in buccal cavity length. The terminal portion of the AZM extends posteriorly in a groove 30–44 7mu; (average = 36.8 μ) on the right side of the body. Dorsally are eight rows of stiff cilia. The details of the silverline system are similar to those of D. scutum .
There is insufficient difference in ranges of body length, width. and buccal cavity length to use these characters in separating the two species. However, a statistical analysis shows that the length of the portion of the AZM on the right side of the body in D. scutum is significantly different (longer) from that of D. peloetes. Furthermore, these two species differ not only in number of dorsal ciliary rows, but also in the number of cilia per row. The degree of difference in these two species is similar to that between closely related species in other hypotrich genera, and also to that between some varieties of Paramecium aurelia.     

Anterior adhesive areas and adjacent secretions in the parasitic flatworms Decacotyle lymmae and D. tetrakordyle (Monogenea: Monocotylidae) from the gills of stingrays

Abstract. The monogeneans Decacotyle lymmae and D. tetrakordyle (Monocotylidae: Decacotylinae), from gills of the dasyatid stingrays Taeniura lymma and Pastinachus sephen , respectively, have a single aperture for adhesive secretion on each side of the anterior ventrolateral region. Rod-shaped bodies (S1) and electron-dense spherical secretion (S2) exit through specialised ducts opening adjacent to one another within these apertures. The S1 bodies are 230 ± 11 nm wide and ≥4 μm long in D. lymmae and 240 ± 9 nm wide and ≥3.3 μm long in D. tetrakordyle . The S2 bodies have a diameter of 88 ± 7 nm in D. lymmae and 65 ± 6 nm in D. tetrakordyle . The apertures are unusual in being extremely small (internal diameter, 3–5 μm). Each aperture has a slit-like surface opening as small as 160 nm wide, surrounded by muscle fibres indicating that they may be opened and closed. The aperture is also surrounded and underlain by muscle fibres that may aid in secretion from, or even eversion of, the tissue within the aperture. Sensilla/cilia are also found within the apertures. Additional secretions from anteromedian and anterolateral glands (body glands), each containing granular secretions, occur in profusion and exit anteriorly and posteriorly to the position of the apertures, through duct openings in the general body tegument. These granular secretions do not appear to be associated with anterior adhesion. Both species show similarities in aperture, underlying tissue, sense organ, and secretion detail, in accordance with findings from other monogenean genera, and which supports the importance of such data for phylogenetic studies.     

Eimeria procyonis sp. n., an Isospora sp., and a Redescription of E. nuttalli Yakimoff and Matikaschwili, 1932 (Protozoa: Eimeriidae) from the American Raccoon (Procyon lotor)

SYNOPSIS. Thirty-two of 48 raccoons examined were infected with a previously undescribed species of Eimeria which is herein named E. procyonis. Of the 32 infected animals, 10 also harbored E. nuttalli and 1 had Isospora sp. oocysts.
The ellipsoid to ovoid oocysts of E. procyonis measured 23.4 × 18.0 (16–29 × 13–24) μm; its sporocysts measured 12.1 × 9.3 (11.5–15 × 7–10) μm, each containing a slightly flattened substiedal body. The sporocyst residuum consisted of numerous scattered granules each ∼1 μm in diameter. The oocyst wall was double-layered. The outer layer appeared rough and pitted, measuring 1.5 μm, except at the micropyle where it was 1 μm thick.
The oocysts of the Isospora sp. measured 16.8 × 13.7 (16–18.5 × 12.5–15.5) μm. The wall consisted of a single layer ∼0.5 μm thick. The sporocysts measured 11.2 × 9.1 (9.5–11.5 × 8–10) μm, and each contained 4 elongate sporozoites. The oocysts of E. nuttalli measured 17.5 × 13.6 (12-21 × 11-15) μm, with a smooth single-layered wall approximately 0.7 μm thick. The sporocysts measured 12.2 × 7.1 (9-13 × 5.5–11) μm. Each sporocyst had a thin, dark, Stieda body and the sporocyst residuum consisted of many fine granules.     

Discovery of Stretch Receptor Organs in Nematodes–Structure, Arrangement and Functional Analysis

Filamentous organs, termed metanemes and interpreted as stretch receptor organs, have been discovered in nematodes of the order Enoplida. They have not been found in species of 11 other nematode orders. According to light microscopical observations, a metaneme consists of a 5–15 μm long scapulus, a 20–400 μm long frontal and a 30–170 μm long caudal cuticularized filament; the latter is lacking in some taxa. Metanemes are arranged serially in the lateral epidermal cords; they are parallel or at an angle of 10–30° to the longitudinal body line. The function of metanemes is likely to coordinate the serpentine movement and to control the actual body volume.     

Actinocephalus carrilynnae N. Sp. (Apicomplexa: Eugregarinorida) from the Blue Damselfly, Enallagma civile (Hagen)

ABSTRACT. Actinocephalus carrilynnae , a new species of actinocephalid gregarine, is described from the blue damselfly, Enallagma civile . Trophozoites are unpaired, lying between the host's gut epithelium and peritrophic membrane, and attain a maximum length of at least 1,700 μ m. Protomerites are subspherical. Epimerites are globular, hemispherical with stub-shaped or truncated cone-shaped projections and are attached to the protomerite by means of a fluted stalk. Protomerite-deutomerite length ratio is 0.12 and relatively constant regardless of trophozoite length. Gametocysts are subspherical, 270–280 μ m in diameter, and undergo sporogenesis in 24–36 h, dehiscing by rupture. Spores are biconical, slightly crescent-shaped, and very uniform in size: 15 μ m long and 4–5 μ m wide. The parasite infects both adult and naiad hosts.     

Two New Species of Isospora from the Desert Skink (Chalchides ocellatus) from the Egyptian Desert

Isospora chalchidis n. sp. and Isospora eimanae n. sp. were isolated from the desert skink, Chalchides ocellatus , order Squamata, from Egypt. Sporulated oocysts of I. chalchidis n. sp. were spheroidal 19 × 19 (18–20.5 × 18–20.5) μm with a 2–layered walled. Oocysts lacked a micropyle and residuum and few polar granules were present. Sporocysts were lemon-shaped 12.2 × 6.5 (9.5–13 × 5–8) μm with a Stieda body and sporocyst residuum but without a substiedal body. Isospora eimanae n. sp. were spheroidal 18.5 × 18.5 (17–19.5 × 17–19.5) μm with a 2–layered wall. There were oocysts without residuum or polar granule, but with small micropyle at 1 end. Sporocysts were ovoid 12 × 8.5(11–13 × 7.5–9) μm with a small Stieda body and sporocyst residuum but without a substiedal body.