Ultrastructure of Tricornia muhezae N. G., N. Sp. (Microspora, Thelohaniidae), a Parasite of Mansonia africana (Diptera: Culicidae) from Tanzania

ABSTRACT. Collections of Mansonia africana mosquito larvae were made at one site in N.E. Tanzania in 1985 and 1987 and from two additional sites, both within about 2 mi of the original one in 1987. An octosporous microsporidian, present at all three sites, was found in both years infecting from 7 to 22% of larvae. Spores (stained in Giemsa) measured 3.0 μ m × 0.25 μ m × 2.25 μ m × 0.26 μ m. Ultrastructurally, spores were seen to have an anterior rim surrounding a depressed area where the endospore was at its thinnest. In transmission electron microscopy section, the rim appeared as two processes into which all layers of the wall extended. At the posterior end all layers of the wall extended into a simple knob-like structure which could be interpreted as a section through a crest running longitudinally around the spore. The polar filament was anisofilar, with two anterior coils of greater diameter than the three posterior coils. Although most closely resembling the genera Amblyspora and Parathelohania in the family Thelohaniidae, the species in M. africana differs from the former, which has oval spores, broadly rounded at the ends, and from the latter, which has a prominent, ridged posterior extension to the spores. The new species has been placed in a new genus and the name Tricornia muhezae proposed.     

Taxonomic Criteria for Limax Amoebae,with Descriptions of 3 New Species of Hartmannella and 3 of Vahlkampfia*

SYNOPSIS. Seven species of limax amoebae were isolated into clonal, monoxenic cultures with Aerobacter aerogenes from material collected from freshwater habitats. Studies were made of their trophic structure, nuclear division, cyst structure, some aspects of cytochemistry, and other characteristics. Six new species are described: Vahlkampfia inornata, V. avara, V. jugosa, Hartmannella limacoides, H. vermiformis, and H. exundans. The well-known species Naegleria gruberi (Schardinger, 1899) is re-described on the basis of 8 strains; its flagellated phase was found to be biflagellate, with rare exceptions. A correlation exists between the manner of locomotion and the pattern of nuclear division in the limax amoebae in the family Vahlkampfiidae and those in the genus Hartmannella. Trophic amoebae of all species had a PAS-positive surface layer, altho results with H. vermiformis and H. exundans were less definite than with other species. All species except H. limacoides formed cysts in culture. The cyst walls of all cyst-forming species were strongly PAS-positive, but results of the zinc chloroiodide test for cellulose were negative with the method used. The genus Hartmannella Alexeieff, 1912, is re-defined to include those species which assume a simple, monopodial limax-like form during locomotion and have nuclear division similar to that of metazoan cells and to distinguish it from the genus Acanthamoeba Volkonsky, 1931.     

A description of seven Antarctic marine gymnamoebae including a new subspecies, two new species and a new genus: Neoparamoeba aestuarina antarctica n. subsp., Platyamoeba oblongata n. sp., Platyamoeba contorta n. sp. and Vermistella antarctica n. gen. n. sp

Seven marine gymnamoebae were isolated from different environments of seawater, slush (pack ice meltwater), and sediment in the Ross Sea area of Antarctica. All amoebae were isolated and maintained at temperatures below 4 degrees C. Growth, rate of locomotion, and general morphology were observed at an environmentally appropriate temperature (1 degrees C) and at room temperature (approximately 25 degrees C). Molecular (srDNA sequences) and microscopical techniques were used to identify the gymnamoebae and establish their phylogenetic affinities. Three isolates (S-131-2, SL-200, and W4-3) were assigned to a psychrophilic subspecies of Neoparamoeba aestuarina, N. aestuarina antarctica n. subsp., one isolate (S-205) was assigned to a new species of Platyamoeba, P. oblongata n. sp., two isolates (W51C#4 & W51C#5) were also assigned to a new species of Platyamoeba, P. contorta n. sp., and one isolate (S-241) was a novel psychrophilic gymnamoeba Vermistella antarctica n. gen. n. sp. Molecular and morphological results revealed that V. antarctica was not related to any described family of gymnamoebae. Strains S-205, W51C#4, and W51C#5 were capable of locomotion at room temperature, while strains SL-200, S-131-2, W4-3, and S-241 exhibited locomotion only below approximately 10 degrees C. Our results imply that the Antarctic environment is host both to cosmopolitan gymnamoebae that have acquired adaptations for existence at low environmental temperature and to apparently novel psychrophilic amoebae described here for the first time.     

Isospora thibetana N. Sp. (Apicomplexa, Eimeriidae), a Parasite of the Tibetan Siskin (Serinus thibetanus=Carduelis thibetanus) (Passeriformes, Fringillidae)

Tibetan siskins are birds native to the Himalayan region often imported into Italy for commercial purposes. Fecal examination of 45 imported subjects with clinical signs of diarrhoea revealed the presence of a large number of coccidian oocysls. After sporulation, accomplished by mixing feces with 2.5 % (w/v) acqueous K₂Cr₂O₇ at room temperature (22° C ± 1° C), exogenous stages of an Isospora species were revealed. The oocysts of this Isospora are spherical, have a bilayered colorless wall, and average 23.24 μm × 23.05 μm; oocyst residuum and micropyle are absent, while an oval polar granule is rarely present. The elliptical sporocysts average 18.44 μm × 10.97 μm and the Stieda body protrudes slightly from the end of the sporocyst. A spherical sporocyst residuum is present though it sometimes consists of scattered granules. The spindle-shaped sporozoites average 11.53 μm × 2.86 μm, and have two refractile bodies. The taxonomic position of the tibetan siskin is controversial. Some authors include this species in the genus Serinus , while others include it in the genus Carduelis. The coccidian species isolated from these tibetan siskins was, for this reason. compared with the Isospora species previously described both in the genus Carduelis and in the genus Serinus. As a result of this comparison a new species. Isospora thibetana , was named. In the intestine of dead subjects, oocysts were found only in the ileum where the mucosa was greatly thickened and presented a heavy leucocytic infiltration consisting mainly of lympho-monocytic cells. A similar infiltration was observed in liver and lungs as well.     

Isospora viridanae N. Sp., an Intranuclear Coccidian Parasite from the Canarian Skink, Chalcides viridanus (Lacertilia: Scincidae)

A caryotropic species of coccidium, Isospora viridanae n. sp. (Apicomplexa: Eimeriidae). has been found as a parasite in the Canarian skink, Chalcides viridanus Gravenhorst, 1851, from Tenerife, Spain, and is described here as a new species. Fully sporulaled oocysts of Isospora viridanae are spherical and measure 21.6 (1 7.6–23.4) nm in diameter. Mtcropyle, oocyst residuum and polar granule are ali absent. Oocysts of this coccidian, with a smooth bilayered wall, contain 2 ovoid sporocysts 13.2 (11.7–14.0) by 9.5 8.2–10 5) μm. A sporocyst residuum is present as well as a Stieda body and a substieda body. Most oocysts are found to be at the beginning of sporulanon when excreted and show 2 spherical sporoblasts. Sporulation is completed within 24 to 48 h at 23 × 2°C Sporozoites are 13–14 μm long and are about 2.5–3 μ m wide. Endogenous stages of schizogony and gamogony develop in the nuclei of epithelial cells from the small intestine of the skink. Comparisons with other species of the genus found in lacertilian hosts indicate that it is a new species.     

Eimeria ornata N. Sp. (Apicomplexa: Eimeriidae) from the Ornate Box Turtle, Terrapene ornata ornata (Reptilia: Testudines), in Texas

Eimeria ornata n. sp. is described from the feces of 6/16 (37.5%) ornate box turtles, Terrapene ornata ornata , in northcentral Texas. Endogenously sporulated oocysts are ellipsoid 17.9 × 15.7(16-21 × 14-18) μm, with a thin, single-layered wall; shape index 1.14 (1.0-1.3). A micropyle is absent but a polar granule was present in one third of the oocysts. An oocyst residuum was present, consisting of numerous small globules situated either in a distinct mass or scattered within the oocyst. The sporocysts are elongate, 11.1 × 5.4 (9-13 × 5-6) μm, with an indistinct Stieda body at 1 pole. A sporocyst residuum is present, consisting either as a compact mass or as scattered granules. The sporozoites are elongate, 9.5 × 2.0 (8-12 × 2) μm, in situ, with spherical anterior and posterior refractile bodies. The new species is distinguished from the similar Eimeria carri Ernst & Forrester, 1973, from eastern box turtles, T. Carolina , by slight differences in oocyst morphology and endogenous sporulation.     

Coccidia of Brazilian Mammals: Eimeria Marajoensis N. Sp. (Apicomplexa: Eimeriidae) From the Anteater, Tamandua Tetradactyla (Xenarthra: Myrmecophagidae)

ABSTRACT Feces from a juvenile specimen of the anteater Tamandua tetradactyla from Ponta de Pedras, Marajó, Pará, northern Brazil, contained three different coccidial oocysts: Eimeria tamanduae Lainson, 1968; E. corticulata Lainson & Shaw, 1990; and a third species previously unrecorded and described here as Eimeria marajoensis n. sp. Oocysts of the latter parasite are spherical to subspherical, 13.9 ± 1.5 times 13.4 ± 1.4 (11.1-16.5 times 11.1-16.5) μm, shape index (length/width) 1.0 (1.0-1.2). the oocyst wall is a single, Colorless layer about 0.6-1.0 μm thick with no striations or micropyle. There is no oocyst residuum, but a single, round, oval or irregularly shaped polar granule of about 0.75-2.5 μm is consistently present. the sporocysts are broadly ellipsoidal, 7.1 ± 0.7 ± 5.3 ± 0.6 (6.0-8.8 times 4.0-5.7) μm, shape index 1.3 (1.2-1.5), with a delicate wall bearing a minute stieda body. No sub-stieda body was visible. the sporocyst residuum consists of some 10-20 rounded granules, lying between the two slightly curved sporozoites which measure approximately 6.5 times 2.0 μm. Sporocyst refractile bodies were not discernablc.     

Heterovesicula cowani N. G., N. Sp. (Heterovesiculidae N. Fam.), a Microsporidian Parasite of Mormon Crickets, Anabrus simplex Haldeman, 1852 (Orthoptera: Tettigoniidae)

ABSTRACT. Heterovesicula cowani , n. g., n. sp., is a dimorphic microsporidium described from the adipose tissue of the Mormon cricket, Anabrus simplex Haldeman. Proliferation of the microsporidium is by karyokinesis of uninucleate and binucleate cells to form binucleate and tetranucleate cells, respectively. These cells will undergo binary fission (merogony). Ultimately, the meronts undergo karyokinesis without subsequent cytokinesis producing spherical multinucleate plasmodia that are transitional to 2 types of sporogony. Transitional to disporoblastic sporogony, a fragile interfacial envelope delaminates from the plasmodium with morphogenesis to a monfiliform plasmodium consisting of fusiform binucleate diplokaryotic sporonts. These undergo karyokinesis to form tetranucleate diplokaryotic sporonts that undergo cytokinesis during disintegration of the plasmodium into isolated binucleate sporonts. Transitional to octosporoblastic sporogony, multinucleate plasmodia disintegrate into short monofiliform plasmodia of diplokaryotic sporonts which then segregate while undergoing gradual nuclear dissociation (haplosis by nuclear dissociation). These undergo two sequences of karyokinesis and subsequent multiple fission to form eight uninucleate (haploid) sporoblasts in a fusiform arrangement within a persistent envelope. Binucleate spores are ovocylindrical, about 5.4 × 1.7 μm (fresh), with an isofilar polar filament singly coiled about 11 turns. Uninucleate spores are ovoid to slightly pyriform, 4.0 × 1.7 μm (fresh), with an isofilar filament singly coiled about 9 turns. A new family, Heterovesculidae, is proposed for the new genus.     

Sphaerospora epinepheli N. Sp. (Myxosporea: Sphaerosporidae) Observed in Grouper (Epinephelus malabaricus)

Sphaerospora epinepheli n. sp. is described from grouper, Epinephelus malabaricus , in cage-cultured and wild fish collected from both coastal lines of southern Thailand. Subspherical to spherical spores and mono- or disporous pseudoplasmodia were observed in the lumen of kidney tubules. Pseudoplasmodia were round to elongate, size range 15.6–22.9 μm (length) × 8.4–21.6 μm (width). Spores were 7.8–10.0 μm (length) × 12.3–14.5 μm (thickness), and 7.0–9.5 μm (width) with two spherical polar capsules of equal size measuring 2.9–4.4 μm in diameter and containing polar filaments with six or seven windings. Two uninucleate sporoplasms showed iodine vacuoles. Blood stages, similar to C-blood protozoans observed from freshwater fish in Europe, were found from peripheral blood smears of grouper. Ultrastructural studies of blood stages showed a similar structure to unidentified mobile protozoans from the blood of carp. Electron dense bodies were observed in the cytoplasm of the primary cell blood stages. Infected proximal-tubular epithelial cells showed highly vacuolated cytoplasm and pycnotic nuclei.     

Isospora atrata N. Sp. (Apicomplexa, Eimeriidae): a New Coccidium Isolated from Carduelis atrata (Passeriformes, Fringillidae)

ABSTRACT Large numbers of coccidian oocysts belonging to the genus Isospora were obtained from the intestinal contents of 98 Carduelis atrata imported into Italy from South America during the months of August through December 1994. The oocysts are sub-spherical and average 21 × 20.3 μm (19.4–23.5 μm × 18.5–22 μm), have a bilayered wall, and an oval polar granule (rarely two). The sporocysts are elliptical and measure 18.8 μm × 10.3 μm (17.5–18.94 μm × 9.5–11.0 μm). The Stieda body protrudes slightly from the end of the sporocyst. A large sporocyst residuum is present, consisting of many granules that may be in a compact mass or scattered. Since this Isospora sp.does not resemble any other species of Isospora previously described from birds of the genus Carduelis , it has been named Isospora atrata n. sp. after the host. Disseminated asexual stages were found in mononuclear cells derived from formalin-fixed post mortem material, suggesting this coccidian may represent an Atoxoplasma -like parasite. Four coccidia-free Serinus canarius L. cohabitated for a long period (4 mo) with infected C. atrata but oocysts were never found in the stool of these birds.     

Abundance of Naked Amoebae in Sediments of Hiroshima Bay, Seto Inland Sea of Japan

ABSTRACT The present paper provides the first data on naked amoebae from sediments of Hiroshima Bay. Three stations in the inner part of the bay were sampled over a three-month period. Abundance of naked amoebae ranged from 1,019 to 45,561 cells/g dry sediment. Results indicate: (i) surface sediment populations in most cases were higher than subsurface populations; (ii) there was some evidence of temporal variation with counts generally increasing from March to May: and (iii) the site located near Hiroshima City had fewer amoebae on several occasions than the other two sites. There was a negative exponential relationship between acid-volatile sulfide concentration and abundance of amoebae. Most amoebae were small with the average size ranging from 6.6–14 μm. Morphotype 1, amoebae that extend lobose pseudopodia or subpseudopodia during normal locomotion, were dominant (40–100% of enumerated amoebae). Morphotypes 2 and 3 (limax amoebae) were found in lower numbers than the other two morphotypes. The proportion of amoebae occupied by Morphotype 4 (fan-shaped or discoidal-flattened amoebae) was higher at a lower total abundance.     

Henneguya adherens N. Sp. (Myxozoa, Myxosporea), Parasite of the Amazonian Fish, Acestrorhynchus falcatus

ABSTRACT. A new species of myxosporean from the gill filaments of the freshwater teleost fish, Acestrorhynchus falcatus collected in the Amazon river is described from light and transmission electron microscope observations. The mature spores (total length 32.3 [30.7–35.1] μ) and all developmental stages were found in the same sporogonic plasmodium. The ellipsoidal spore body consists of 2 unequal shell valves adhering together along the suture lines. Each valve, tapering as a caudal projection, forms a long tail (length 20.5 [18.0–21.7] μm). The tail was surrounded by a homogeneous sheath on its length. The polar capsules measuring 3.1 × 1.2 μm contain 3–4 coils of the polar filament. All surfaces of the immature and mature spores were surrounded by a closely adherent homogenous structural sheath, mainly thicker around the tails. The taxonomic affinities of this parasite to other species are discussed.     

Learamoeba waccamawensis, N. G., N. Sp. (Heterolobosea: Vahlkampfiidae), a New Temperature-Tolerant Cyst-Forming Soil Amoeba

Soil from a roadside ditch at Lake Waccamaw, North Carolina, was tested for cyst-forming free-living amoebae, and water from the same ditch was tested for fecal coliform bacteria. Soil samples incubated at room temperature (21-23° C) yielded Acanthamoeba polyphaga, Amoeba sp., Hyperamaeba sp., Mayorella penardi. Naegleria gruberi, Naegleria minor , and unidentified ciliates, testaceans and slime molds. Incubation at 37-39° C yielded Acanthamoeba jacobsi, Platyamoeba schaefferi , and a new amoeba to be described herein. Fecal contamination of the ditch was confirmed by a direct membrane filtration technique that yielded a mean of 1,410 + 134 bacteria/ ml. The new amoeba is assigned to the class Heterolobosea Page & Blanton, 1985 on the basis of promitotic nuclear division, and a flagellated stage with a cytostome as seen in the genus Tetramitus. Amoebae studied in hanging drop preparations were flattened and irregular as described for the genus Stachyamoeba in the family Gruberellidae but changed to a lobose cylindrical form as described for species of Vahlkampfia in the family Vahlkampfiidae. A new genus and species, Learamoeba waccamawensis , is proposed for the amoeba described herein.     

Pellitidae n. fam. (Lobosea, Gymnamoebia) – a new family, accommodating two amoebae with an unusual cell coat and an original mode of locomotion, Pellita catalonica n.g., n.sp. and Pellita digitata comb. nov

Two species of amoebae, the marine Pellita catalonica n. g., n. sp. discovered in the Ebro Delta (Spain) and Nivå Bay (Denmark) and the freshwater Pellita digitata n. comb., previously known from the UK and Switzerland and now found in North-Western Russia, have a very thick (0.5–0.8 μm) cell coat consisting of a fuzzy fibrous basal layer, covered with the dense layer of complex pentagonal glycostyles. The cell coat is integrated with the cell membrane and entirely envelops the amoeba, like a typical glycocalyx. For purposes of locomotion and phagocytosis the cell produces short papilliform subpseudopodia that protrude through the cell coat and are covered solely by the cell membrane. In order to accommodate these unusual organisms we established a new family, Pellitidae n. fam., within the subclass Gymnamoebia sensu Page, 1987, order Euamoebida.     

Wittmannia antarctica N. G., N. Sp. (Nosematidae), a New Hyperparasite in the Antarctic Dicyemid Mesozoan Kantharella antarctica

ABSTRACT. Collections of the dicyemid mesozoan Kantharella antarctica were made in the Weddell Sea during the Antarctic Expedition of the research vessel RV Polarstern in 1990 and 1991. A diplokaryotic microsporidian was found infecting all nematogens from all the samples taken in both years. The infected cells contained all developmental stages. Merogony initially was monokaryotic and spoorogony of diplokaryotic sporonts was by multiple fission. The stained ovoidal spores measured between 4.3-6 μm X 1.7-2.3 μm. The ultrastructural findings come from 11 specimens of Kantharella antarctica that were cut in serial sections. All developmental stages were noteworthy because of the myelinosomes situated adjacent to each diplokaryon. Similarly conspicuous were some organelles in the spore: a prominent, extraordinarily electron dense anterior portion of the polaroplast and the posterior vacuole. The isofilar polar filament with a diameter of about 115 nm showed 9-11 coils. The great number of empty spore cases together with an extruded polar filament are indicative of an autoinfection. Though these characteristics resemble in part those of the genus Nosema from the family Nosematidae, the species in Kantharella antarctica differs from the former by its unusual development, life cycle and unusual host. Thus, this new species has been placed in a new genus and the name Wittmannia antarctica proposed.     

Isospora elmahalensis n. sp. (Apicomplexa, Eimeriidae), a Parasite of the White-Cheeked Bulbul (Pycnonotus leucogenys) in Saudi Arabia

Isospora elmahalensis n. sp. is described from the Saudi Arabian bird, Pycnonotus leucogenys , from the Elmahala valley. Sporulated oocysts of I. elmahalensis were spherical or nearly subspherical, 19.5–22.5 × 18.5–20 (21.34 ± 0.4 × 19.06 ± 0.5) μm. Oocysts lacked a micropyle, residuum, and polar granule. Sporocysts were ovoid, 14–17.5 × 7–12 (16.08 ± 1.05 × 9.9 ± 1.55) μm, and had a Stieda body and sporocyst residuum, but lacked a substiedal body. Sporozoites were elongated with a clear globule at one end. The host bird belongs to the order Passeriformes.     

Eimeria turcicus N. Sp. (Apicomplexa: Eimeriidae) from the Mediterranean Gecko, Hemidactylus turcicus (Sauria: Gekkonidae)

ABSTRACT. Eimeria turcicus n. sp. (Apicomplexa: Eimeriidae) is reported from the gall bladder of the Mediterranean gecko, Hemidactylus t. turcicus (Linnaeus, 1758) from the Houston Zoo, Texas, USA. Oocysts of this coccidian are elongate and cylindrical, 38.2 × 17.9 (35.2-40.8 × 16.8-20.0)μm, with a smooth, bilayered wall ∼ 1.6 μm thick; shape index 2.1 (1.9-2.3). A polar granule is present, but a micropyle and oocyst residuum are absent. Sporocysts are ovoid, 11.0 × 8.8 (10.0-12.0 × 8.0-9.4) μm, with a smooth, thin wall and suture; shape index 1.3 (1.1-1.4). Each sporocyst contains a residuum measuring 6.0 × 5.1 (4.8-8.0 × 4.8-6.4) μm, additional residual granules scattered among the sporozoites, and two sporozoites that are normally arranged head-to-tail within the sporocyst. Each sporozoite contains a single, ovoid, posterior refractile body and a central nucleus.     

Nadelspora canceri N. G., N. Sp., an Unusual Microsporidian Parasite of the Dungeness Crab, Cancer Magister

ABSTRACT. The microsporidium Nadelspora canceri n. g., n. sp., is described from the striated musculature of the Dungeness crab ( Cancer magister ) in Oregon, USA. The needle-shaped spores were rounded anteriorly, tapered to a posterior point and measured 7.1–11.8 × 0.2–0.3 μm in fixed preparations. The extremely narrow spore diameter prevented observation of morphological details at the light microscopic level and ultrastructural details of mature spores were difficult to resolve. Meronts were not observed and the monokaryotic merozoites and sporonts were not contained within either parasitophorous or sporophorous vesicles. Sporonts were disporoblastic and gave rise to monokaryotic sporoblasts that became narrow and elongate as they developed into immature spores with a developing polar filament. The nucleus was not clearly resolved in mature spores and may have been surrounded by the lamellar polaroplast. The polar filament was of nearly uniform diameter throughout most of its length and ended abruptly about three-fourths of the distance from the anterior end of the spore. Unusual spherical non-membrane bound granules surrounded the polar filament in a spiral arrangement. The new microsporidium resembles members of the family Mrazekiidiae, but differs in lacking a diplokaryon at any stage. It is probably most closely related to Baculea daphniae from which it differs primarily by spore shape and size. The familial relationships of the genus Baculea have not been determined and it is proposed to include it with Nadelspora in the new family Nadelsporidae.     

Fine Structure of the Feeding Stage of A Sorogenic Ciliate, Sorogena Stoianovitchae Gen. N., Sp. N.

SYNOPSIS. A new species of kinetophragminophoran ciliate, collected from dried vegetation and capable of forming an aerial sorocarp, is described and named Sorogena stoianovitchae gen. n., sp. n. This ciliate is a voracious predator that feeds on species of Colpoda , and, when the latter is depleted in numbers, aggregates to forms sorogens. Each sorogen rises into the air from the surface of the water, forming a secreted stalk with a sorus of cysts at its apex. the feeding stage of the ciliate resembles an Enchelys in that it has an apical, slit-like mouth surrounded by a lip, a somewhat dorso-ventrally flattened body, and meridional kineties. Its length ranges from 40–75 μm and width from 23–55 μm. It has a typical rhabdos type of cytopharynx, but no specialized oral ciliature. the somatic kineties are formed of rows of paired kinetosomes with associated microfibrils, the arrangement of which differs a little from that of other ciliates of this subclass. Sorogena has tentatively been placed in the order Haptorida although it lacks toxicysts, recognizable mucocysts, and clavate cilia. Its unique life cycle and some of the details of its fine structure indicate differences between Sorogena and other haptorids so profound that a new family, SOROGENIDAE, is created for it. the type species (PNG76-73) was collected on dry figs at the Wau Ecology Institute, Papua New Guinea.