Euplotespora binucleata n. gen., n. sp. (Protozoa: Microsporidia), a parasite infecting the hypotrichous ciliate Euplotes woodruffi, with observations on microsporidian infections in ciliophora

A new microsporidian species, Euplotespora binucleata n. gen., n. sp., from the brackish-water ciliate Euplotes woodruffi is described and defined on the basis of life history characteristics, light and electron microscopic features, and small subunit (SSU) ribosomal DNA (rDNA) sequencing. The life cycle of E. binucleata n. sp. probably has rather short merogonic and relatively long sporogonic phases. Some uninuclear meronts and sporonts, along with diplokaryotic sporoblasts and spores, were found in experimentally infected host cells. Such a peculiar life cycle has been induced experimentally in Euplotes eurystomus and constitutively microsporidian-free stocks of E. woodruffi. Spores of E. binucleata n. sp. are monomorphic, ovoid-cylindrical in shape, 3.44+/-0.17 x 1.65+/-0.22 microm in size, and characterized by a diplokaryotic condition and a large posterior vacuole. The polar tube is isofilar, 4.5-5.5 microm in length when ejected, and lacking a distinctive coiled region (half-coiled). The polaroplast is divided into two regions: the anterior part has a few lamellae close to the anchoring disc; and the posterior part is a rounded body (sack), about one-quarter of the spore length. Spores do not appear to cluster together as a group. Each spore is surrounded by a sporophorous membrane closely adjacent to the exospore layer. A phylogenetic analysis of SSU rDNA sequences by different methods placed E. binucleata n. sp. in a clade with representatives of the microsporidian genera Cystosporogenes and Vittaforma. Observations of microsporidia in several other ciliates are discussed in view of the microsporidian infection frequency in the phylum Ciliophora.     

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.     

Norlevinea n. g., a New Genus for Glugea daphniae (Protozoa: Microspore), a Parasite of Daphnia longispina (Crustacea: Phyllopoda)1

ABSTRACT. Norlevinea n. g. is established for microsporidia in which a uninucleate meront changes into a sporont by secreting a thin, membranous, sporontogcnetic and fragile sporophorous vesicle (pansporoblast membrane) in which four uninucleate sporoblasts are formed. In contrast to the genus Gurleya, the sporoblasts and later the spores are permanently joined into doublets, being laterally cemented by an electron-dense substance structurally identical to and continuous with the exospore layer. The polar filament is of the anisofilar type. The type species is Norlevinea daphniae (Weiser, 1947) n. comb., a parasite of the ovaries of Daphnia longispina occurring in several carp ponds in Czechoslovakia.     

Amazonspora hassar n. gen. and n. sp. (phylum Microsporidia,fam. Glugeidae), a parasite of the Amazonian teleost Hassar orestis (fam. Doradidae)

We describe the microsporidian Amazonspora hassar n. gen., n. sp. from the gill xenomas of the teleost Hassar orestis (Doradidae) collected in the estuarine region of the Amazon River. The parasite appeared as a small whitish xenoma located in the gill filaments near the blood vessels. Each xenoma consisted of a single hypertrophic host cell (HHC) in the cytoplasm of which the microsporidian developed and proliferated. The xenoma wall was composed of up to approximately 22 juxtaposed crossed layers of collagen fibers. The plasmalemma of the HHC presented numerous anastomosed, microvilli-like structures projecting outward through the 1-3 first internal layers of the collagen fibrils. The parasite was in direct contact with host cell cytoplasm in all stages of the cycle (merogony and sporogony). Sporogony appears to divide by plasmotomy, giving rise to 4 uninucleate sporoblasts, which develop into uninucleate spores. The ellipsoidal spores measured 2.69 +/- 0.45 x 1.78 +/- 0.18 microm, and the wall measured approximately 75 nm. The anchoring disk of the polar filament was subterminal, being shifted laterally from the anterior pole. The polar filament was arranged into 7-8 coils in a single layer in the posterior half of the spore, surrounding the posterior vacuole. The polaroplast surrounded the uncoiled portion of the polar filament, and it was exclusively lamellar. The spores and different life-cycle stages were intermingled within the cytoplasm of the HHC, surrounding the central hypertrophic deeply branched nucleus. The ultrastructural morphology of this microsporidian parasite suggests the erection of a new genus and species.     

Microfilum lutjani N. G. N. Sp. (Protozoa Microsporida), a gill parasite of the golden African snapper Lutjanus fulgens (Valenciennes, 1830) (Teleost Lutjanidae): developmental cycle and ultrastructure

Microfilum lutjani n. g., n. sp. (Microsporida) was found on the gill filaments of Lutjanus fulgens (Teleost) inhabiting the coasts of Senegal. This microsporidium forms xenomas distinguished by the microvilli covering the plasma membrane. At all stages of development individuals have isolated nuclei and are in direct contact with the host cytoplasm. Merogony is binary and sporogony is tetrasporoblastic. The spore (4.75 x 2.60 microns) is characterized by a manubrium inserted on a laterally offset anchoring disc and extending into a very short, noncoiled polar filament (no longer than 500 nm) in the form of a hook. This type of polar filament has not been described previously in the Microsporida.     

Microfilum Lutjani N. G. N. Sp. (Protozoa Microsporida), A Gill Parasite of the Golden African Snapper Lutjanus Fulgens (Valenciennes, 1830) (Teleost Lutjanidae): Developmental Cycle and Infrastructure

ABSTRACT Microfilum lutjani n. g., n. sp. (Microsporida) was found on the gill filaments of Lutjanus fulgens (Teleost) inhabiting the coasts of Senegal. This microsporidium forms xenomas distinguished by the microvilli covering the plasma membrane. At all stages of development individuals have isolated nuclei and are in direct contact with the host cytoplasm. Merogony is binary and sporogony is tetrasporoblastic. the spore (4.75 times 2.60 μm)) is characterized by a manubrium inserted on a laterally offset anchoring disc and extending into a very short, noncoiled polar filament (no longer than 500 nm) in the form of a hook. This type of polar filament has not been described previously in the Microsporida.     

Pyrotheca hydropsycheae N. Sp., a Microsporidian Parasite of Caddis Fly Larvae,Hydropsyche siltalai Döhler, 1963 (Trichoptera,Hydropsychidae)1

ABSTRACT. Pyrotheca hydropsycheae n. sp. is described from caddis fly larvae, Hydropsyche siltalai Döhler, 1963. All stages were found in oenocytes and fat body cells. Meronts were uni- or binucleate with simple surface membranes. The sporogonic stages were recognized ultrastructurally by the separation of an envelope, the sporophorous vesicle, from their surfaces. Mature sporogonial plasmodia were tetranucleate and gave rise by longitudinal fission to four uninucleate elongate sporoblasts with polar nuclei. Spores were lageniform with an inflated posterior end, containing the polar tube coils and the nucleus, and a narrow anterior section comprising two-thirds of the length, containing the polaroplast and straight part of the polar tube. The polaroplast consisted of an anterior region of loosely packed membranes arranged as partitions at angles to one another and a posterior region of increasingly closely packed parallel membranes. The spore wall consisted of an electron-dense exospore with a fuzzy coat and a thin electron-lucent endospore. All four spores derived from a sporont faced in the same direction in the sporophorous vesicle. Spores measured 8.7 μm long and extruded polar filaments were about 20 μm.     

Fine structure of the microsporidan Abelspora portucalensis gen.n., sp.n. (Microsporida) parasite of the hepatopancreas of Carcinus maenas (Crustacea,Decapoda)

A new species of a microsporidan, Abelspora portucalensis, was found in the hepatopancreas of Carcinus maenas, forming white xenomas. Each xenoma seems to consist of an aggregate of hypertrophic host cells in which the parasite develops and proliferates. This cytozoic microsporidan being characterized by one uninucleate schizont giving rise to two sporonts, each originating two sporoblasts, resulting in two spores within a persistent sporophorous vacuole (pansporoblast) should be included in a new family Abelsporidae. In fresh smears most spores were 3.1–3.2 μm long and 1.2–1.4 μm wide. Fixed, stained, and observed in SUS mature spores measured 3.1 ± 0.08 × 1.3 ± 0.06 μm (n = 25 measurements). Spore cytoplasm was dense and granular, polyribosomes were arranged in helicoidal tape form. The polar filament was anisofilar and consisted of a single coil with 5–6 turns. The anchoring disc and and the anterior zone of the filament are surrounded by the polaroplast composed of two usual zones. In the anterior zone, the membrane of the polar filament is in continuity with the membranes of the polaroplast. The appearance of a microsporidan with described nuclear divisions in life cycle, spores shape and size, polaroplast and polar filament morphology and identity of the host suggests that we may erect a new genus Abelspora and a new species A. portucalensis (Portugal = Portucalem).     

Description of Hyalinocysta expilatoria n. sp., a microsporidian parasite of the blackfly Odagmia ornata

Hyalinocysta expilatoria n. sp. is described from a larva of Odagmia ornata collected in Sweden. Infection was restricted to the adipose tissue which was transformed into a syncytium. The earliest stage observed was diplokaryotic merozoites, which mature directly into diplokaryotic sporonts. Each sporont produces a sporophorous vesicle (pansporoblast), which persists, also enclosing mature spores. Usually nuclear divisions result in a plasmodium with 8 nuclei, which fragments into 8 sporoblasts, each of which develops into a spore without further division. Occasionally an aberrant number of spores (2, 4, 6) is formed. The spores are pyriform with a flattened area at the posterior pole. Spores in sporophorous vesicles with 8 spores are 4.0–6.0 μm long, in vesicles with 4 spores 4.0–5.0 μm, and in vesicles with 2 spores 7.0–8.0 μm. In some vesicles the spores develop asynchronously, and 2, 4, or 6 mature spores are found together with 6, 4, or 2 immature. There was also a small number of vesicles with supernumerary spores, less than 8 normally developed. The 325–350 nm thick spore wall is composed of three layers. The polar filament is anisofilar with 7 coils in a single layer. The anterior 5–6 coils are wide, the posterior 2-1 thin. The angle of tilt of the anterior filament coil is approximately 50°. The spore has a single nucleus. The sporophorous vesicle is delimited by a thin membrane, also visible in haematoxylin stained preparations. Vesicles with mature spores are void of metabolic inclusions.     

Occurrence of a new microsporidan: Enterocytozoon bieneusi n.g., n. sp., in the enterocytes of a human patient with AIDS

A new microsporidium is reported infesting the enterocytes of a Haitian patients with AIDS. The stages observed were diplokaryotic cells, sporogonial plasmodia, unikaryotic sporoblasts, and spores. Neither a sporophorous vesicle (pansporoblastic membrane) nor parasitophorous vacuole were differentiated around the developmental stages, which were in direct contact with the host cell cytoplasm. The polar tube (5-6 coils) was differentiated before fission of the sporogonial plasmodium. The mature spores measured 1.5 micron X 0.5 micron. The spore wall was very thin as the endospore was absent or poorly differentiated. The organism is named Enterocytozoon bieneusi n. g., n. sp. and is assigned to the suborder Apansporoblastina.     

Occurrence of a New Microsporidan: Enterocytozoon bieneusi n. g., n. sp., in the Enterocytes of a Human Patient with AIDS1

A new microsporidium is reported infesting the enterocytes of a Haitian patient with AIDS. The stages observed were diplokaryotic cells, sporogonial plasmodia, unikaryotic sporoblasts, and spores. Neither a sporophorous vesicle (pansporoblastic membrane) nor parasitophorous vacuole were differentiated around the developmental stages, which were in direct contact with the host cell cytoplasm. The polar tube (5-6 coils) was differentiated before fission of the sporogonial plasmodium. The mature spores measured 1.5 m?m × 0.5 m?. The spore wall was very thin as the endospore was absent or poorly differentiated. The organism is named Enterocytozoon bieneusi n. g., n. sp. and is assigned to the suborder Apansporoblastina.     

Glugea gasti sp. n., a Microspridan Pathogen of the Boll Weevil Anthonomus grandis1,2,3

SYNOPSIS. Glugea gasti sp. n., a microsporidan pathogen of Anthonomus grandis Boheman (the boll weevil), is described and a probable life cycle presented. The alimentary canal, and probably the mesenteron 1st, is the initial site of infection, altho the disease later becomes generalized thruout most body tissues. Binucleate sporoplasms initiate the 1st schizogonic phase, characterized by mono- and bi-nucleate schizonts. The 2nd schizogonic phase is characterized by mono-, bi- and quadrinucleate schizonts, by prolific multiplication, by the dense compact nuclei early in this phase, and late in this phase by larger schizonts with less dense vesicular nuclei. This phase terminates in formation of diplokarya. The sporogonic phase is characterized by combination of the 2 nuclei in the diplokaryon followed by nuclear divisions in a sequence closely resembling meiosis. Two sporoblasts are produced from each sporont. Mature spores in wet mounts by phase contrast were 4.3 ± 0.3 μ long by 2.3 ± 0.2 μ wide. The polar filament averaged 76 μ long. Mature spores were present about 24 hours after infection. Some observations are presented on an external filament extending from one pole of the spore to host tissue and other events during the process of spore morphogenesis.     

Microgemma vivaresi n. sp. (Microsporidia, Tetramicridae), infecting liver and skeletal muscle of sea scorpions, Taurulus bubalis (Euphrasen 1786) (Osteichthyes, Cottidae), an inshore, littoral fish

The ultrastructure of a new microsporidian species Microgemma vivaresi n. sp. causing liver cell xenoma formation in sea scorpions, Taurulus bubalis, is described. Stages of merogony, sporogony, and sporogenesis are mixed in the central cytoplasm of developing xenomas. All stages have unpaired nuclei. Uninucleate and multinucleate meronts lie within vacuoles formed from host endoplasmic reticulum and divide by binary or multiple fission. Sporonts, no longer in vacuoles, deposit plaques of surface coat on the plasma membrane that cause the surface to pucker. Division occurs at the puckered stage into sporoblast mother cells, on which plaques join up to complete the surface coat. A final binary fission gives rise to sporoblasts. A dense globule, thought to be involved in polar tube synthesis, is gradually dispersed during spore maturation. Spores are broadly ovoid, have a large posterior vacuole, and measure 3.6 microm x 2.1 microm (fresh). The polar tube has a short wide anterior section that constricts abruptly, then runs posteriad to coil about eight times around the posterior vacuole with granular contents. The polaroplast has up to 40 membranes arranged in pairs mostly attached to the wide region of the polar tube and directed posteriorly around a cytoplasm of a coarsely granular appearance. The species is placed alongside the type species Microgemma hepaticusRalphs and Matthews 1986 within the family Tetramicridae, which is transferred from the class Dihaplophasea to the class Haplophasea, as there is no evidence for the occurrence of a diplokaryotic phase.     

Ultrastructural aspects of a new species,Vavraia mediterranica (Microsporidia,Pleistophoridae), parasite of the French Mediterranean shrimp,Crangon crangon (Crustacea,Decapoda)

The life cycle stages of a new species of the genus Vavraia (Microsporidia, Pleistophoridae), which parasitizes the shrimp Crangon crangon (Crustacea, Decapoda), were examined by light and electron microscopy. This parasite was monomorphic with polysporous sporogony and developed in the skeletal muscle of the host. The multinucleate sporogonial plasmodium divided by plasmotomy and multiple division into uninucleate sporoblasts. All stages were surrounded by a thick and amorphous dense coat external to the plasmalemma. This structure gradually became a merontogenetic sporophorous vacuole (MSV) where the sporonts developed into sporoblasts. The MSV was filled with episporontal granular secretory products and eventually contained up to 50 uninucleate spores. During spore morphogenesis, these episporontal granular products within the MSV became organized as episporontal tubular-like structures. In transverse sections, these structures showed a mean diameter of 1.0 microm, but disappeared during the final phase of the spore maturation. Mature spores were ellipsoidal to slightly pyriform and measured 2.30 x 1.41 microm. The polar filament was anisofilar and consisted of a single coil with six to seven turns (rarely five). This new species is named Vavraia mediterranica n. sp.     

Ultrastructure and Description of a New Species of Telomyxa (Microspora: Telomyxidae) from the Semiaquatic Beetle,Ora texana Champ. (Coleoptera: Helodidae)1

A new species of the uncommon microsporidian genus Telomyxa (Microspora: Telomyxidae) has been found parasitizing the larval fat body of the semiaquatic beetle, Ora texana. In this species, the sporogonic sequence results in the formation of sporocysts measuring 7.7 times 6.5 μm that contain two crested uninucleate spores (averaging 5.7 times 2.2 μm). The spores are essentially oblong/ovate, tapering toward the anterior end and remaining bound together after sporogony by a persistent accessory membrane or sporocyst. The two spores in the sporocyst are produced by an unusual morphogenetic sequence in which, after one mitosis, the binucleate sporont elongates, forming two lobes that fold toward one another and cleave along a central plane, forming two parallel sporoblasts. The general ultrastructural features of this process are described, and diagnostic characters of this new species of Telomyxa are presented.     

Pleistophora finisterrensis n. sp., a microsporidian parasite of blue whiting Micromesistius poutassou

Pleistophora finisterrensis n. sp. is a microsporidian parasite of the hypoaxial musculature of the blue whiting Micromesistius poutassou (Risso). Foci of infection are between 3 and 6 mm in length and have no evident effects on adjacent muscle fibres. We found only a single type of spore (uninucleate, with mean dimensions of 4×2 µm in fresh preparations), contained within sporophorous vesicles (mean diameter 19 µm in fresh preparations; 150–250 spores per vesicle). All of the development stages of this microsporidian are monokaryotic. The meronts are initially uninucleate and bounded by a plasmalemma. Towards the end of merogony, meronts are multinucleate plasmodia with a well-defined surface coat. Sporogony is polysporous, with multinucleate sporonts, which likewise have a well-defined surface coat (about 130 nm thick), dividing by plasmotomy to give rise to uninucleate sporoblasts. The polar tube is isofilar and consists of 8–9 turns in the posterior half of spore. The polaroplast is made up of an anterior lamellar part and a posterior vesicular part.     

Development, ultrastructure, natural occurrence, and molecular characterization of Liebermania patagonica n. g., n. sp., a microsporidian parasite ot the grasshopper Tristira magellanica (Orthoptera: Tristiridae)

A new microsporidium, Liebermannia patagonica n. gen., n. sp., is described from midgut and gastric caecum epithelial cells of Tristira magellanica, an apterous grasshopper species of southern Patagonia, Argentina. L.patagonica is diplokaryotic, apansporoblastic, homosporous, and polysporoblastic. Transitional (from merogony to sporogony) stages and sporonts of L. patagonica were surrounded by host rough endoplasmic reticulum. The ovocylindrical spores measured 2.9 +/- 0.09 x 1.2 +/- 0.04 microm (fresh, n = 50), and they had an isofilar polar filament of only three coils and a cluster of tubules instead of a classical posterior vacuole. Prevalence was high (up to 80.6%) at the type locality for the four years sampled . Maximum likelihood , neighbor joining, maximum parismony analyses of the small submit rDNA all placed L.patagonica(Accession No. DQ 239917) in one with Orthosomella operophterae.     

Kabataia gen. n., a new genus proposed for Microsporidium spp. infecting trunk muscles of fishes

Based on a fine structural study, a new genus, Kabataia gen. n., is proposed for Microsporidium arthuri Lom, Dyková and Shaharom, 1990. It develops in trunk muscles of a South-East Asian freshwater fish, Pangasius sutchi. The genus has nuclei isolated throughout the cycle, merogony stages are multinucleate, sporogony proceeds in 2 steps: multinucleate sporont segments into sporoblast mother cells which produce 2 sporoblasts. Sporoblasts and early spores are characterized by a dense globule at the site of the posterior vacuole. Mature spores are of a rather variable shape. Their exospore is raised into small, irregular fields. The polaroplast is relatively small and its posterior part consists of flat vesicles with dense contents. The polar tube makes a small number (4 to 6) of turns. A congeneric species is Kabataia seriolae (Egusa, 1982) comb. nov. from cultured marine yellowtails Seriola quinqueradiata. Kabataia inflicts heavy damage on muscle tissue. The sarcoplasm within which Kabataia develops is reduced to an amorphous mass with tubule-like fibrils, microfibrils and small vesicles.     

Ultrastructural study on a novel microsporidian, Endoreticulatus eriocheir sp. nov. (Microsporidia, Encephalitozoonidae), parasite of Chinese mitten crab, Eriocheir sinensis (Crustacea, Decapoda)

A microsporidian pathogen, infecting the epithelial cells of the hepatopancreas of Chinese mitten crab, Eriocheir sinensis, was studied by electron microscopy. The detailed ultrastructure of life cycle of the pathogen including proliferative and sporogonic developmental stages are provided. All stages of the parasite are haplokaryotic and develop in a vacuole bounded by a single membrane in contact with host cell cytoplasm. Sporogenesis is synchronous with the same developmental stage in one vacuole. Sporogony shows a characteristic of multinucleate sporogonial plasmodia divided by rosette-like division, producing 4 or 8 sporoblasts. The mature spore is ellipsoidal, length (mean) 1.7 microm, width 1.0 microm, with a uninucleate in the center of the sporoplasm, 7 turns of the polar filament, a bell-like polaroplast of compact membranes and obliquely positioned posterior vacuole. The morphological characteristics of this novel microsporidian pathogen have led us to assign the parasite to a new species of Endoreticulatus, E. eriocheir sp. nov., that has not been reported previously from crab.