Brachiola vesicularum, N. G., N. Sp., a New Microsporidium Associated with AIDS and Myositis

Brachiola vesicularum, n. g., n. sp., is a new microsporidium associated with AIDS and myositis. Biopsied muscle tissue, examined by light and electron microscopy, revealed the presence of organisms developing in direct contact with muscle cell cytoplasm and fibers. No other tissue types were infected. All parasite stages contain diplokaryotic nuclei and all cell division is by binary fission. Sporogony is disporoblastic, producing 2.9 times 2 μm diplokaryotic spores containing 8-10 coils of the polar filament arranged in one to three rows, usually two. Additionally, this microsporidium produces electron-dense extracellular secretions and vesiculotubular appendages similar to Nosema algerae. However, the production of protoplasmic extensions which may branch and terminate in extensive vesiculotubular structures is unique to this parasite. Additionally, unlike Nosema algerae , its development occurred at warm blooded host temperature (37-38° C) and unlike Nosema connori , which disseminates to all tissue types, B. vesicularum infected only muscle cells. Thus, a new genus and species is proposed. Because of the similarities with the genus Nosema , this new genus is placed in the family Nosematidae. Successful clearing of this infection (both clinically and histologically) resulted from treatment with albendazole and itraconozole.     

Transfer of Nosema locustae (Microsporidia) to Antonospora locustae n. comb. based on molecular and ultrastructural data

Nosema locustae is a microsporidian parasite of grasshopper pests that is used as a biological control agent, and is one of the emerging model systems for microsporidia. Due largely to its diplokaryotic nuclei, N. locustae has been classified in the genus Nosema, a large genus with members that infect a wide variety of insects. However, some molecular studies have cast doubt on the validity of certain Nosema species, and on the taxonomic position of N. locustae. To clarify the affinities of this important insect parasite we sequenced part of the rRNA operon of N. locustae and conducted a phylogenetic analysis using the complete small subunit rRNA gene. Nosema locustae is only distantly related to the nominotypic N. bombycis, and is instead closely related to Antonospora scoticae, a recently described parasite of bees. We examined the ultrastructure of mature N. locustae spores, and found the spore wall to differ from true Nosema species in having a multi-layered exospore resembling that of Antonospora (one of the distinguishing features of that genus). Based on both molecular and morphological evidence, therefore, we propose transferring N. locustae to the genus Antonospora, as Antonospora locustae n. comb.     

Morphological and molecular investigations of Tubulinosema ratisbonensis gen. nov., sp. nov. (Microsporidia: Tubulinosematidae fam. nov.), a parasite infecting a laboratory colony of Drosophila melanogaster (Diptera: Drosophilidae)

A new species of microsporidia from Drosophila melanogaster was investigated by light and electron microscopy and by ribosomal RNA (rRNA) sequencing. This microsporidium and the previously described Nosema kingi and Nosema acridophagus have been transferred to the new genus Tubulinosema gen. nov. with the following characters: nuclei are in diplokaryotic arrangement during the life cycle. All stages are in direct contact with the host cell cytoplasm, slightly anisofilar polar tube with the last coils being smaller in diameter arranged in one or two rows on both sides of the diplokaryon and small tubuli on the surface of late meronts. Spores are oval or slightly pyriform. Thick endospore wall, thinner over anchoring disc. This new genus and the genus Brachiola have been placed in a new family Tubulinosematidae fam. nov. Phylogenetic analysis of small subunit rRNA sequences by different methods placed Tubulinosema spp. in one clade with the genus Brachiola forming its sister clade, which is distant from the clade containing the true Nosema spp. including Nosema bombycis.     

Establishment of the new genus Paranosema based on the ultrastructure and molecular phylogeny of the type species Paranosema grylli Gen. Nov., Comb. Nov. (Sokolova, Selezniov, Dolgikh, Issi 1994), from the cricket Gryllus bimaculatus Deg

The ultrastructure of the microsporidian parasite Nosema grylli, which parasitizes primarily fat body cells and haemocytes of the cricket Gryllus bimaculatus (Orthoptera, Gryllidae) is described. All observed stages (meront, meront/sporont transitional stage ("second meront"), sporont, sporoblast, and spore) are found in direct contact with the host cell cytoplasm. Nuclei are diplokaryotic during almost all stages of the life cycle, but a brief stage with one nucleus containing an abundance of electron-dense material is observed during a "second merogony." Sporogony is disporous. Mature spores are ovocylindrical in shape and measure 4.5+/-0.16micromx2.2+/-0.07 microm (n=10) on fresh smears and 3.3+/-0.06 micromx1.4+/-0.07 microm (n=10) on ultrathin sections. Spores contain 15-18 coils of an isofilar polar filament arranged in one or two layers. Comparative phylogenetic analysis using rDNA shows N. grylli to be closely related to another orthopteran microsporidian, Nosema locustae, and to Nosema whitei from the confused flour beetle, Tribolium confusum. Antonospora scoticae, a parasite of the communal bee Andrena scotica, is a sister taxon to these three Nosema species. The sequence divergence and morphological traits clearly separate this group of "Nosema" parasites from the "true" Nosema clade containing Nosema bombycis. We therefore propose to change the generic name of N. grylli and its close relative N. locustae to Paranosema n. comb. We leave N. whitei in former status until more data on fine morphology of the species are obtained.     

Nosema algerae n. sp. (Cnidospora, Microsporida) a Pathogen in a Laboratory Colony of Anopheles stephensi Liston (Diptera, Culicidae)

SYNOPSIS. A new microsporidan, Nosema algerae n. sp., was found in a laboratory colony of Anopheles stephensi. The microsporidan infects a variety of tissues of both larvae and adults and is highly pathogenic for its host. The structure and life cycle of the microsporidan under the light and electron microscope is described and its relationship to other Nosema species infecting mosquitoes is discussed.     

Description of Scalithrium n. gen. (Cestoda,Tetraphyllidea) with Scalithrium minimum (Van Beneden, 1850) n. comb., a parasite of Dasyatis pastinaca (Elasmobranchii,Dasyatidae), as type species

Scalithrium gen. n. (Cestoda, Tetraphyllidea) is proposed with Scalithrium minimum (Van Beneden, 1850) n. comb., parasite of Dasyatis pastinaca (Elasmobranchii, Dasyatidae) as type-species. The new genus Scalithrium (Tetraphyllidea, Phyllobothriidae, Rhinebothriinae) is erected for several species previously included in the genus Rhinebothrium. These species have a scolex with four bothridia, the distal surface of which is divided by transverse septa in a single row of loculi. Scalithrium minimum (Van Beneden, 1850) n. comb. is redescribed from specimens collected from the type-host Dasyatis pastinaca in Tunisia and becomes the type-species of the new genus. After Braun (1900) Echeneibothrium variabile Van Beneden, 1850 is considered as type-species of the genus Echeneibothrium. Species of Rhinebothriinae to be transferred into the genus Scalithrium are discussed and a key is proposed for the eight species.     

Studies on ocular microsporidia.

Sera from six ocular microsporidiosis patients and eight individuals with no history of microsporidiosis were assayed by enzyme-linked immunosorbent assay (ELISA) and by Western blot immunodetection. Microsporidia used as antigen include Nosema corneum, Encephalitozoon hellem, Encephalitozoon cuniculi, and Nosema algerae. Three AIDS patients with known E. hellem infections displayed ELISA antibody titers to E. hellem ranging from 1:400 to 1:12,800. Two patients with unclassified microsporidial infections displayed highest antibody titers to N. algerae (1:1,600 and 1:3,200), a mosquito microsporidian which, reportedly, cannot infect man. A sixth patient with a known N. corneum infection displayed the same ELISA antibody titer (1:1,600) to all four microsporidia. Western blot patterns also were variable among the patient sera; however, the most intense and complex antibody-binding patterns corresponded with the higher ELISA antibody titers. Sera from eight HIV-seronegative individuals with no history of microsporidiosis reacted variably to the four microsporidia. These results suggest that diagnosis of microsporidiosis may depend upon direct detection of the organisms using species-specific antibodies or molecular probes rather than conventional serology.     

Development of Edhazardia aedis (Kudo, 1930) N. G., N. Comb. (Microsporida: Amblyosporidae) in the Mosquito Aedes aegypti (L.) (Diptera: Culicidae)

A microspondium of the mosquito Aedes aegypti (L.), identified as Nosema aedis Kudo, 1930, was found to be a heterosporous species with 3 sporulation sequences. Usually, I sequence developed in a parental generation host individual that was infected per os as a larva and the other 2 developed concurrently in a filial host larva that was infected transovarially. Under some conditions there were deviations from the parental host-filial host alternation. The 1st sporulation sequence was diplokaryotic (diploid in a particular sense) throughout; the other 2 arose from diplokaryotic meronts, developed concurrently and ended with haploid spores. Haplosis in 1 case was by means of dissociation of the diplokaryon. In the other case it was by meiosis. Conflicting reports about whether the members of the diplokaryon in the latter sequence separate and undergo meiosis individually or coalesce and undergo meiosis as I nucleus were resolved in favor of the latter idea. A new genus in family Amblyosporidae was created to contain this species. which then became Edhazardia aedis (Kudo. 1930) n. g., n. comb.     

Development of Edhazardia aedis (Kudo, 1930) n. g., n. comb. (Microsporida: Amblyosporidae) in the mosquito Aedes aegypti (L.) (Diptera: Culicidae)

A microsporidium of the mosquito Aedes aegypti (L.), identified as Nosema aedis Kudo, 1930, was found to be a heterosporous species with 3 sporulation sequences. Usually, 1 sequence developed in a parental generation host individual that was infected per os as a larva and the other 2 developed concurrently in a filial host larva that was infected transovarially. Under some conditions there were deviations from the parental host-filial host alternation. The 1st sporulation sequence was diplokaryotic (diploid in a particular sense) throughout; the other 2 arose from diplokaryotic meronts, developed concurrently and ended with haploid spores. Haplosis in 1 case was by means of dissociation of the diplokaryon. In the other case it was by meiosis. Conflicting reports about whether the members of the diplokaryon in the latter sequence separate and undergo meiosis individually or coalesce and undergo meiosis as 1 nucleus were resolved in favor of the latter idea. A new genus in family Amblyosporidae was created to contain this species, which then became Edhazardia aedis (Kudo, 1930) n. g., n. comb.     

Nosema chrysorrhoeae n. sp. (Microsporidia), isolated from browntail moth (Euproctis chrysorrhoea L.) (Lepidoptera, Lymantriidae) in Bulgaria: characterization and phylogenetic relationships

A new microsporidian parasite Nosema chrysorrhoeae n. sp., isolated in Bulgaria from the browntail moth (Euproctis chrysorrhoea L.), is described. Its life cycle includes two sequential developmental cycles that are similar to the general developmental cycles of the Nosema-like microsporidia and are indistinguishable from those of two Nosema spp. from Lymantria dispar. The primary cycle takes place in the midgut tissues and produces binucleate primary spores. The secondary developmental cycle takes place exclusively in the silk glands and produces binucleate environmental spores. N. chrysorrhoeae is specific to the browntail moth. Phylogenetic analysis based on the ssu rRNA gene sequence places N. chrysorrhoeae in the Nosema/Vairimorpha clade, with the microsporidia from lymantriid and hymenopteran hosts. Partial sequences of the lsu rRNA gene and ITS of related species Nosema kovacevici (Purrini K., Weiser J., 1975. Natürliche Feinde des Goldafters, Euproctis chrysorrhoea L., im Gebiet von Kosovo, FSR Jugoslawien. Anzeiger fuer Sch?dlingskunde, Pflanzen-Umweltschutz, 48, 11-12), Nosema serbica Weiser, 1963 and Nosema sp. from Lymantria monacha was obtained and compared with N. chrysorrhoeae. The molecular data indicate the necessity of future taxonomic reevaluation of the genera Nosema and Vairimorpha.     

The ultrastructure of three microsporidia from winter moth,Operophtera brumata (L.), and the establishment of a new genus Cystosporogenes n.g. for Pleistophora operophterae (Canning, 1960)

Summary The ultrastructure of three species of microsporidia in winter moths, Operophtera brumata (L.), has been used to consolidate taxonomic assessments previously based on light microscopy. The characters formerly used to assign Nosema operophterae Canning, 1960 to a new genus Orthosoma Canning, Wigley & Barker, 1983, namely that the nuclei are isolated and that sporoblasts are separated from ribbon-shaped multinucleate (2, 4, 8 or rarely 12 nuclei) sporonts, were upheld at the ultrastructural level. Development was in contact with the cell cytoplasm but all stages, which must have included meronts, had an electron dense surface coat. Nosema wistmansi Canning, Wigley & Barker, 1983, was found to be ultrastructurally typical of the genus Nosema Naegeli, 1857. An unusual feature of this species was the close association of cysternae of host endoplasmic reticulum with the surface of meronts, an association lost in sporogony. Pleistophora operophterae (Canning, 1960) has been transferred, on ultrastructural criteria, to a new genus Cystosporogenes n.g. Nuclei are isolated; all stages develop in a vesicle bounded by an envelope of enigmatic origin; this envelope persists around the spores as a sporophorous vesicle; division of the sporont within this vesicle is by budding and the number of sporoblasts, and therefore spores, is variable up to about 60.Microsporidia which undergo multisporous sporogony in sporophorous vesicles are now distributed among seven genera. These are: Glugea Thélohan, 1891; Pleistophora Gurley, 1893; Pseudopleistophora Sprague, 1977; Vavraia Weiser, 1977; Baculea Loubès & Akbarieh, 1978; Polydispyrenia Canning & Hazard, 1982 and Cystosporogenes n.g. New genera would appear to be needed for Pleistophora sp. of Sandars & Poinar (1976) and Pleistophora sp. of Percy, Wilson & Burke (1982). ac]19840404     

Small Subunit Ribosomal DNA Phylogeny of Various Microsporidia with Emphasis on AIDS Related Forms

ABSTRACT. Phylogenetic analysis of the small subunit ribosomal DNA of a broad range of representative microsporidia including five species from humans ( Enterocytozoon bieneusi, Nosema corneum, Septata intestinalis, Encephalitozoon hellem and Encephalitozoon cuniculi ), reveals that human microsporidia are polyphyletic in origin. Septata intestinalis and E. hellem are very similar to the mammalian parasite E. cuniculi . Based on the results of our phylogenetic analysis, we suggest that S. intestinalis be designated Encephalitozoon intestinalis . Furthermore, analysis of our data indicates that N. corneum is much more closely related to the insect parasite Endoreticulatus schubergi than it is to other Nosema species. This finding is supported by recent studies which have shown a similarity between E. schubergi and N. corneum based on the origin and development of the parasitophorous vacuole. Thus these opportunistic microsporidian parasites can originate from hosts closely or distantly related to humans. Finally, the phylogeny based on small subunit ribosomal DNA sequences is highly inconsistent with traditional classifications based on morphological characters. Many of the important morphological characters (diplokaryon, sporophorous vesicle, and meiosis) appear to have multiple origins.     

On the hoplonemertean, Kameginemertes parmiornatus (Iwata, 1957) gen. n., comb. n. from Sagami Bay, Japan

The hoplonemertean Amphiporus parmiornatus Iwata,1957, from Sagami Bay, Japan, has beenredescribed and named Kameginemertes gen. n. asa member of the reptantic Polystilifera. Anatomicallythe new genus shows affinity to the genus Drepanophorus belonging to the EureptantiaAequifurcata, but because of differences in thecerebral sensory organs, nervous system,blood-vascular system, and rhynchodaeal wall, a newgenus is proposed for it.     

Microsporidian Parasites of Trematode Larvae from Aquatic Snails in West Malaysia

In a survey of microsporidian parasites of trematode larvae of Malaysian fresh-water snails, species of Nose-matidae were found 7 times in rediae from 425 Lymnaea rubiginosa, 6 times in rediae from 97 Indoplanorbis exustus and not at all in other snails. There were 3 species of Microsporida in all: one, found in Echinoparyphium dunni, Echinostoma hystricosum and Echinostoma malayanum was identified as Nosema eurytremae Canning; another, which was found in E. dunni and Echinostoma audyi, had paired nuclei (diplokaryon form) in sporogony only and was named a new species, Nosema vasicola sp.n.; the 3rd, from E. hystricosum, lacked diplokaryon nuclei throughout development, was considered to belong to a new genus and was named Unikaryon piriformis gen.n., sp.n. Nosema eurytremae was transmitted experimentally to Fasciola hepatica rediae in Lymnaea truncatula.     

Nosema helminthorum Moniez, 1887 (Microspora,Nosematidae):A Taxonomic Enigma1

ABSTRACT. The microsporidian parasite known as Nosema helminthorum Moniez, 1887, parasitic in the tapeworm Moniezia expansa (Rudolphi, 1810), has been shown by electron microscopy to have two cycles of development, one with isolated nuclei, the other with paired nuclei (diplokarya). Both merogony and sporogony of the two separate sequences take place in direct contact with the host cell cytoplasm and ultimately give rise to unikaryotic and diplokaryotic sporoblasts. Sporogony is disporoblastic. The nuclear condition of the spores was not seen. The sequences, corresponding to those of the genera Unikaryon and Nosema, may be part of a single dimorphic life cycle and, if so, the species will have to be transferred to a new genus.     

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.     

Description of Oculogryphus shuensis sp. n. (Coleoptera,Lampyridae), the first species of the genus in the Sino-Japanese realm,with a modified key to the subfamily Ototretinae

A new species of the lampyrid genus Oculogryphus Jeng, Engel, and Yang, O. shuensis sp. n. from China (Sichuan Province) is described and figured. The genus previously was known only from Vietnam, and the new species is the first representative of the genus in the Sino-Japanese zoogeographic realm. Some morphological variations of Oculogryphus and the allied genus Stenocladius are discussed and a modification to the most recent key to ototretine genera is proposed to accommodate Oculogryphus.     

Morphological and molecular studies of a microsporidium (Nosema sp.) isolated from the thee spot grass yellow butterfly, Eurema blanda arsakia (Lepidoptera: Pieridae)

A microsporidium possessing molecular and morphological characteristics of the genus Nosema was isolated from larvae of the thee-spot grass yellow butterfly, Eurema blanda arsakia. The complete rRNA gene sequences of the E. blanda isolate contained 4,428 base pairs (GenBank Accession No. EU338534). The organization of the rRNA genes is LSU rRNA-ITS-SSU rRNA-IGS-5S, which corresponds with that of Nosema species closely related to Nosema bombycis. Phylogenetic analysis based on rRNA gene sequences show that this isolate is closely related to Nosema bombycis, Nosema plutellae, Nosema spodopterae, and Nosema antheraeae. The ultrastructure of all developmental stages of this microsporidium confirmed its placement in the genus Nosema. The isolate was successfully propagated in cell lines IPLB-LD652Y (Lymantria dispar) and NTU-LY (Lymantria xylina) and, in the in vitro system, it was frequently found to develop in the nuclei of the host cells, a circumstance that seldom occurs in other Nosema species. An extra-cellular vegetative stage of this microsporidium was also observed in the culture medium after 14 days of infection. The ECMDFs might be released from disrupted host cells.     

Cinetozona pyriformis n. g., n. sp.: a Relative of the Ciliate Genera Urozona and Cinetochilum (Ciliophora, Scuticociliatida)

Cinetozona pyriformis n. g., n. sp. is a very small scuticociliate recently found in a freshwater artificial pond in Madrid, Spain. This new ciliate is here described using silver impregnation techniques. C. pyriformis bears a girdle of cilia near the equator ot the cell, similar to the genus Urozona Schewiakoff, 1889, while the oral structures resemble those in the genus Cinetochilum Perty, 1852. The ciliate's systematic position is discussed, and its inclusion in the family Cinetochilidae is proposed.