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.     

Molecular and ultrastructural characterization of Andreanna caspii n. gen., n. sp. (Microsporida: Amblyosporidae), a parasite of Ochlerotatus caspius (Diptera: Culicidae)

A new genus and species of microsporidia, Andreanna caspii n. gen., n. sp. is described from the mosquito, Ochlerotatus caspius (Pallas) based on ultrastructural morphology, developmental characteristics, and comparative sequence analyses of the small subunit (SSU) ribosomal DNA (rDNA). Parasite development is confined to fat body tissue and infected larvae appear swollen with dull white masses within the thorax and abdomen. Meronts have diplokaryotic nuclei and are delineated by a simple plasmalemma contiguous with the host cell cytoplasm. Merogony occurs by synchronous binary division followed by cytokinesis. Diplokaryotic sporonts undergo meiosis and synchronous nuclear division forming sporogonial plasmodia with two, four and eight nuclei enclosed within a persistent sporophorous vesicle. Cytokinesis of sporogonial plasmodia results in the formation of eight uninucleate spores. The episporontal space of early sporonts is filled with a homogeneous accumulation of electron dense granular inclusions and ovoid vesicles of various dimensions, transforming into an interwoven matrix during the initial phase of sporogenesis. Spores are oval, uninucleate and measure 4.8 ± 0.3 × 3.1 ± 0.4 μm (fixed). The spore wall is 260 μm thick with an irregular exospore consisting of two layers (150-170 μm) and a thinner endospore (90-100 μm). The anchoring disk is well developed and is contiguous with a lamellar polaroplast that occupies the anterior third of the spore and possess more narrow lamellae on the posterior end. The polar filament is gradually tapered and arranged in a single row consisting of six coils ranging from 180 to 150 μm in diameter. The posterior vacuole (posterosome) is moderately sized and filled with a matrix of moderate electron density. Phylogenetic analysis of SSU rDNA from A. caspii and 30 other species of microsporidia including 11 genera parasitic in mosquitoes using maximum parsimony, neighbor joining and maximum likelihood methods showed A. caspii to be a sister group to the clade containing all of the Amblyospora species, including Culicospora, Edhazardia and Intrapredatorus, as well as Culicosporella and Hyalinocysta thus providing strong support for establishment of Andreanna as a separate genus.     

Ultrastructure ofNosema marucae sp. n. (Microspora,Nosematidae), a pathogen ofMaruca testulalis (Lepidoptera,Pyralidae)

The development ofNosema marucae was followed in its host, the legume pod borerMaruca testulalis. The meronts had an irregular cell body and were binucleated, with the large diplokaryon occupying most of the cell. The sporonts contained many rough endoplastic reticula. The sporoblasts had a granular cytoplasm with a large number of ribosomes, and nuclear division and cellular cleavage were prevalent, particularly from 84 to 120 h after inoculation. Sporoblasts and young spores were evident from 108 to 144 h after inoculation.In the mature spores, the exocuticle had a rough and sculptured surface and the endocuticle thinned considerably at the apical end. The anchoring disc was thick at the point of attachment. The polar tube had 12–15 loops, and the polaroplast was multilayered with two distinct regions and occupied almost half of the spore. The diplokaryon was centrally located in the cell; the nuclear membranes of the diplokarya were separated by an electron-dense region about 0.6 nm thick.     

Amblyospora sp. (Microspora, Amblyosporidae) infecting nerve ganglia of Culex pipiens (Diptera, Culicidae) from Egypt.

A species of Amblyospora-infecting neurones of Culex pipiens is described. Diplokaryotic meronts, which divided by binary fission, were distinguished at the electron microscope level by their unthickened plasma membranes. Sporonts with an electron-dense surface coat gave rise to eight uninucleate sporoblasts within a sporophorous vesicle, cytoplasmic division occurring at the quadrinucleate or octonucleate stages. Indications that nuclear fusion and chromosome reorganization occurred in merogony and sporogony were obtained by light microscopy but meiosis was not detected at the ultrastructural level. Spores were typical of Amblyospora, being ovoid when fresh, truncate when stained, and having an exospore of two membranous layers subtended by a thick amorphous layer, an electron-lucent endospore, an anisofilar polar filament, and a polaroplast comprised of an anterior region of close-packed lamellae and a posterior region of expanded sacs. The metabolic products in the sporophorous vesicle took the form of large globules, small globules with electron-dense borders, and fine granules. These were depleted in mature sporophorous vesicles, though a surface layer of fine granules on the spores may have been derived from them. Many stages were degenerate and it is suggested that C. pipiens may be an accidental host in which the parasite could develop suboptimally in nervous tissue only. Infections in larvae hatched from eggs in the laboratory indicate that vertical transmission occurs.     

Saetheriella amplicristata gen. n., sp. n., a new Orthocladiinae (Diptera: Chironomidae) from Tennessee

The generic diagnosis of the male imago of a new genus of the subfamily Orthocladiinae (Chironomidae), Saetheriella is given. The genus is characterized by protruding, hairy eyes; antenna with straight, apical seta; antepronotum reduced; squama without setae; Cu₁ distinctly downcurved; and anal point short, triangular, covered with microtrichia and with two strong, lateral setae. The genus is close to Gynnidocladius Sublette & Wirth, Unniella Sæther and Parakiefferiella Thienemann. The distribution of the genera shows evidence of a Gondwanian connection. The male imago of the only included species S. amplicristata sp. n. is described.     

Ultrastructure of Buddenbrockia allmani n. sp. (Myxozoa, Malacosporea), a parasite of Lophopus crystallinus (Bryozoa, Phylactolaemata)

Development of a new species of malacosporean myxozoan (Buddenbrockia allmani n. sp.) in the bryozoan Lophopus crystallinus is described. Early stages, represented by isolated cells or small groups, were observed in the host's body wall or body cavity. Multiplication and rearrangement of cells gave an outer cell layer around a central mass. The outer cells made contact by filopodia and established adherens junctions. Sporoplasmosomes were a notable feature of early stages, but these were lost in subsequent development. Typical malacosporean sacs were formed from these groups by attachment of the inner (luminal) cells by a basal lamina to the outer layer (mural cells). Division of luminal cells gave rise to a population of cells that was liberated into the lumen of the sac. Mitotic spindles in open mitosis and prophase stages of meiosis were observed in luminal cells. Centrioles were absent. Detached luminal cells assembled to form spores with four polar capsules and several valve cells surrounding two sporoplasms with secondary cells. Restoration of sporoplasmosomes occurred in primary sporoplasms. A second type of sac was observed with highly irregular mural cells and stellate luminal cells. A radially striated layer and dense granules in the polar capsule wall, and previous data on 18 rDNA sequences enabled assignment of the species to the genus Buddenbrockia, while specific diagnosis relied on the rDNA data and on sac shape and size.     

On the Cytology and Taxonomic Position of Nudispora biformis N. G., N. Sp. (Microspora, Thelohaniidae), a Microsporidian Parasite of the Dragon Fly Coenagrion hastulatum in Sweden

The microsporidium Nudispora biformis n. g., n. sp., a parasite of a larva of the damsel fly Coenagrion hastulatum in Sweden, is described based on light microscopic and ultrastructural characteristics. Merogonial stages and sporonts are diplokaryotic. Sporogony comprises meiotic and mitotic divisions, and finally eight monokaryotic sporoblasts are released from a lobed plasmodium. Sporophorous vesicles are not formed. The monokaryotic spores are oval, measuring 1.4–1.8 × 2.8–3.4 μm in living condition. The thick spore wall has a layered exospore, with a median double-layer. The polaroplast has two lamellar parts, with the closest packed lamellae anteriorly. The isofilar polar filament is arranged in 6 (to 7) coils in the posterior half of the spore. Laminar and tubular extracellular material of exospore construction is present in the proximity of sporogonial stages. In addition to normal spores teratological spores are produced. The microsporidium is compared to the microsporidia of the Odonata; its possible relations to the genus Pseudothelohania and to the Thelohania-like microsporidia are discussed. The new genus is provisionally included in the family Thelohaniidae.     

Significance of transovarial infections of Amblyospora sp. (Microspora:Thelohaniidae) in relation to parasite maintenance in the mosquito Culex salinarius

Adult females of Culex salinarius, transovarially infected with the microsporidium Amblyospora sp., showed no significant differences in overall fecundity, physiological longevity, and preoviposition periods when compared to healthy adults under laboratory conditions. Development times and survival rates for congenitally infected young to reproductive age were also indistinguishable from those of healthy controls. A significant reduction of 52% in egg hatch was observed for infected eggs when compared to healthy eggs. Prevalence rates of infection for progeny produced by infected females declined with each successive gonotrophic cycle and averaged 90%. Transovarial transmission is not sufficient for the maintenance of the microsporidium in a population of mosquitoes. An alternate host is suggested as a mechanism whereby the microsporidium can reenter a healthy mosquito population.     

Bilatorchis papillogenitalis n.g., n.sp. (Paramphistomidae: Orthocoelinae), a parasite of the red lechwe (Kobus leche Gray, 1850) from Zambia

Summary Bilatorchis papillogenitalis n.g., n.sp. from the red lechwe, Kobus leche Gray, 1850, from Zambia is described and illustrated. It is assigned to the subfamily Orthocoelinae Price & McIntosh, 1953 and distinguished from other members of the subfamily by the lateral and symmetrical position of the testes in the posterior third of the body between the caecal ends and the acetabulum, the intertesticular position of the ovary and Mehlis' gland and the limited extent of the caeca which end only at the level of the anterior border of the testes. A new type of genital atrium, papillogenitalis type is proposed for B. papillogenitalis, Calicophoron papillosum, Carmyerius parvipapillatus and, possibly, Orthocoelium parvipapillatum. The genus Cochinocotyle Gupta & Gupta, 1970 is moved to the subfamily Orthocoelinae.Supported by a scholarship grant under the 4th Education Project of the Philippines with the World Bank.Supported by a scholarship grant under the 4th Education Project of the Philippines with the World Bank.     

Crepidula beklemishevi gen. et sp. n. and Dimeiospora palustris gen. et sp. n. (Microspora: Amblyosporidae)--new microsporidian genera and species from blood-sucking mosquitoes (Diptera: Culicidae) from the south of the western Siberia

Microsporidia parasitizing the adipose body of mosquito larvae of Anopheles beklemishevi and Aedes punctor has been studied. Two new genera of microsporidia are described based on lightmicroscopic and ultrastructural characteristics of spores and sporogony stages. The spore wall of Crepidula beklemishevi gen. n. et sp. n. is formed by two-membrane exospore, thick exospore, bilayer endospore and thin plasmolemma. Spores with single nucleus, polar filament anisofilar, with 6-7 coils (2+ 4-5), polaroplast consisting of three parts: macrochelicoidal, microhelicoidal and lamellar. Fixed spores 4.2 +/- 0.22 x 2 +/- 0.01 microns. The sporogony of Dimeiospora palustris gen. et. n. results in spore formation of two different types. Spores of the first type are oviform, with thick wall, single-nuclear, 6.1 x 4.9 microns. Spore wall with three layers, about 370 nm. Exospore electron-dense, subexospore moderately electrondense. Exospore and subexospore irregularly pleated on the almost spore surface and slightly thinner on anterior end only. Endospore electron-translucent. Polar filament anisofilar, with 9 coils (3 + 6). Polaroplas consists of three parts: lamellar, fine bubbled, and coarse bubbled. Spores of the second type broad-ovate, with apical pole narrower, distal pole concave, 4.6 x 3.7 microns. Spore wall with three layer, 355 nm. Exospore on the apical end irregularly pleated, consists of thin electrondense exospore, subexospore of variable electron density, endospore electron-translucent. Polar filament anisofilar, with 13 coils (3 + 10). Polaroplast has two parts: lamellar and vesicular.     

Description of Auriculotrema lechneri n. gen., n. sp. (Digenea: Choanocotylidae), a parasite of freshwater turtles (Testudines: Pleurodira: Chelidae) from Queensland,Australia

Auriculotrema lechneri n. gen., n. sp. is described from the small intestine of Emydura krefftii and Elseya latisternum from northern Queensland, Australia. The new species strongly resembles species of Choanocotyle in body shape, ventral incision of the oral sucker, structure of the cirrus sac, and location of the genital pore. The distinctive taxonomic feature is the presence of 2 winglike projections extending beyond the lateral margins of the oral sucker, in contrast to the extremely large, expanded oral sucker diagnostic of Choanocotyle spp. Auriculotrema n. gen. is the second genus included in the formerly monotypic Choanocotylidae Jue Sue and Platt, 1998.     

Leptoconops nosopheris sp. n. (Diptera: Ceratopogonidae) and Paleotrypanosoma burmanicus gen. n., sp. n. (Kinetoplastida: Trypanosomatidae), a biting midge--trypanosome vector association from the Early Cretaceous

Leptoconops nosopheris sp. n. (Diptera: Ceratopogonidae) is described from a blood-filled female biting midge in Early Cretaceous Burmese amber. The new species is characterized by a very elongate terminal flagellomere, elongate cerci, and an indistinct spur on the metatibia. This biting midge contained digenetic trypanosomes (Kinetoplastida: Trypanosomatidae) in its alimentary tract and salivary glands. These trypanosomes are described as Paleotrypanosoma burmanicus gen. n., sp. n., which represents the first fossil record of a Trypanosoma generic lineage.     

Mizelleus chauhani, n. sp., a branchial parasite of Wallago attu (Bloch)

The authors describe a new species of the genus Mizelleus Jain, 1957. Its characteristic features include the number of head organs, the presence of pharyngeal glands, the shape of the copulatory complex, the structure of the egg and the shape of the dorsal transverse bar, the ventral hamuli and the ventral transverse bar.     

Notes on Empididae (9), Heleodromia haenii sp.n., an Aquatic Empidid (Diptera) from France

Heleodromia haenii sp.n., a new aquatic dance fly (Diptera, Empididae, Hemerodromiinae) from France is described and figured. It is a member of the subgenus Illiesiella and readily distinguished from its congeners by the possession of S‐shaped, upright distal part of the periandrium.     

Riethia manauara n. sp., an Amazonian Chironomid (Diptera: Chironomidae) from Brazil

One new species of Riethia Kieffer, Riethia manauara n. sp., is described and figured as male, pupa and larva. The generic diagnosis for pupae and larvae are emended. The specimens were collected from water systems in the Amazon Rainforest in northern Brazil.