Stempellia milleri sp. n. (Microsporida: Nosematidae) in the Mosquito Culex pipiens quinquefasciatus Say*

Stempellia milleri sp. n. was found in blood cells and the adipose tissue of field-collected larvae of Culex pipiens quinquefasciatus Say. Its development is different from other Stempellia species described in that it is dimorphic, producing two types of spores in adipose cells. Both were studied with the electron microscope. One spore is thin-walled and uninucleate; the other is thick-walled and binucleate. The latter spores are produced from sporonts that are somewhat similar to those of Nosema. Sporoblasts and spores of S. milleri were compared to those of Stempellia magna Kudo, and Stempellia lunata Hazard and Savage in both light and electron microscope preparations. S. milleri was transmitted experimentally to Culex pipiens pipiens, C. p. quinquefasciatus, C. salinarius, C. tarsalis, and C. territans. Transmission of S. magna to its host and other mosquito species was not possible. Although, generally low numbers of test larvae became infected with the pathogen, heavy infections were seen occasionally in individual specimens of its natural host, C. p. quinquefasciatus. Species of Aedes, Anopheles, Culiseta, Psorophora, and Uranotaenia exposed to spores of S. milleri were not susceptible to the disease.     

Amblyospora trinus N. Sp. (Microsporida: Amblyosporidae) in the Australian Mosquito Culex halifaxi (Diptera: Culicidae)

ABSTRACT. A new species of Amblyospora , a parasite found in wild populations of the predacious Australian mosquito Culex halifaxi , was investigated with light and electron microscopy. This species was found to be heterosporous with two concurrent sporulation sequences in the host larvae, both arising from diplokaryotic meronts and ending with haploid spores. One sequence was dominant and involved meiosis to produce eight thick-walled, broadly oval meiospores in a sporophorous vesicle (SV). The other sequence involved nuclear dissociation to produce lanceolate, thin-walled spores in a subpersistent SV. Horizontal transmission to the mosquito host, by one or both of two distinctly different pathways (one via an intermediate host, the other by cannibalism of infected individuals) and by vertical transmission, are postulated but have not been demonstrated. A new species, Amblyospora trinus , is proposed and its affinities to other heterosporous microsporidia in mosquitoes are discussed.     

Ultrastructural and Cytochemical Observations on Sporogenesis of Myxobolus sp. (Myxosporida: Myxobolidae) from the Common Shiner Notropis cornutus*

SYNOPSIS. The structure and cytochemistry of spores of Myxobolus sp. from plasmodia which occur in the gill filaments of the common shiner Notropis cornutus were studied by light microscopy and by scanning and transmission electron microscopy. The thin-walled valves of the pyriform spores are thickened in the lateral sutural and apical regions. Mucous material is associated predominantly with the posterior end of many spores. The plasmodium is surrounded by a syncytial wall bounded by 2 membranes. Pinocytotic channels are formed by the inner membrane and numerous dense vesicles are pinched off at the distal ends of the channels. Sporogenesis is initiated by the envelopment of one vegetative cell by another. The larger, enveloped cell divides to form a disporous pansporoblast, which contains 2 pairs of capsulogenic and valvogenic cells and 2 binucleate sporoplasm cells. Each capsular primordium and connecting external tubule gives rise to a polar capsule which houses a helically coiled polar tubule. The apical end of each polar capsule is plugged by a stopper. The valvogenic cells surround the capsulogenic and posteriorly situated sporoplasm cells to form the spore valves. Iodinophilic (glycogen) inclusions were not seen in spores stained with iodine or Best's carmine. A darkly stained band was observed around the posterior region of most spores stained with Best's carmine. In the electron microscope large aggregates of β glycogen particles were seen in the cytoplasm of sporoplasm cells in mature spores.     

Ultrastructural Study of the Encystation and Excystation Processes in Naegleria sp.

ABSTRACT. An important aspect of the biology of Naegleria sp. is the differentiation processes that occur during encystation and excystation. We studied these using both fluorescence and transmission electron microscopy techniques. In the initial stages of encystation, the cisternae of the endoplasmic reticulum became densely filled with a fibrillar material. Vesicles with a similar content that appeared to be derived from the cisternae were also observed in close contact with the plasma membrane. As encystation progressed, the fibrillar material became localized on the surface of the amoeba. An irregular compaction was observed in some areas of the cyst wall, which contained thin extensions of the cyst wall fibrillar material. Completely formed cysts had two to three ostioles, each sealed by an operculum. The operculum contained two areas in which a differential compaction of the fibrillar structure was observed. When excystation was induced, small dense granules (DGs), which were in close contact with fibrillar material were observed in the cyst cytoplasm and in the peritrophic space. During excystation, the more compact component of the operculum moves to enable the pseudopod of the emerging trophozoite to penetrate the ostiole. Vacuoles containing a fibrillar material, probably derived from the cyst wall, were observed in the cytoplasm of the pseudopodia. Our results provide a platform for further studies using biochemical markers to investigate the origin of the cyst wall as well as the role of DGs during excystation in Naegleria .     

Hyalophysa Iwoffi sp.n. from the Fresh-water Shrimp Palaemonetes paludosus and Revision of the Genus Hyalophysa

SYNOPSIS. Hyalophysa Iwoffi sp. n. is described from the shrimp, Palaemonetes paludosus . Its life cycle is like that of other exuviotrophic apostomes. Hyalophysa Iwoffi differs from the type species, H. chattoni , in the details of its infraciliature and the shape of its macronucleus during the trophont stage. Emended diagnoses of the genus, Hyalophysa , and the species, H. chattoni , also are given.     

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.     

Functional morphology of the muscles in Philodina sp. (Rotifera: Bdelloidea)

Whole-mounts of Philodina sp., a bdelloid rotifer, were stained with fluorescent-labeled phalloidin to visualize the musculature. Several different muscle types were identified including incomplete circular bands, coronal retractors and foot retractors. Based on the position of the larger muscle bands in the body wall, their function during creeping locomotion and tun formation was inferred. Bdelloid creeping begins with the contraction of incomplete circular muscle bands against the hydrostatic pseudocoel, resulting in an anterior elongation of the body. One or more sets of ventral longitudinal muscles then contract bringing the rostrum into contact with the substrate, where it presumably attaches via adhesive glands. Different sets of ventral longitudinal muscles, foot and trunk retractors, function to pull the body forward. These same longitudinal muscle sets are also used in `tun' formation, in which the head and foot are withdrawn into the body. Three sets of longitudinal muscles supply the head region (anterior head segments) and function in withdrawal of the corona and rostrum. Two additional pairs of longitudinal muscles function to retract the anterior trunk segments immediately behind the head, and approximately five sets of longitudinal retractors are involved in the withdrawal of the foot and posterior toes. To achieve a greater understanding of rotifer behavior, it is important to elucidate the structural complexity of body wall muscles in rotifers. The utility of fluorescently-labeled phalloidin for the visualization of these muscles is discussed and placed in the context of rotifer functional morphology.     

Phylogenetic distance of Thelohania butleri Johnston, Vernick, and Sprague, 1978 (Microsporidia; Thelohaniidae), a parasite of the smooth pink shrimp Pandalus jordani, from its congeners suggests need for major revision of the genus Thelohania Henneguy, 1892

Thelohania butleri, a microsporidian that causes mortality and commercial losses in the smooth pink shrimp Pandalus jordani, is of taxonomic interest as a species resembling the poorly studied type species, Thelohania giardi, of the large, polyphyletic genus Thelohania. We examined the ultrastructure of T. butleri to confirm its identity and reconstructed phylogenies using ribosomal DNA to find the relationship of T. butleri with other Thelohania species in crayfish and ants. Light and transmission electron microscopy from specimens collected from the type locality, the Pacific coast of Canada, confirmed the identity and demonstrated a development similar to that of T. giardi, involving a series of binary fissions without formation of a plasmodium. Phylogenetic analyses consistently showed T. butleri to be distantly related to other Thelohania species, and closely related to species from marine decapods within a larger fish-parasitic clade. Together, features such as host group and habitat, developmental morphology, and phylogeny suggest T. butleri may be a closer relative to T. giardi than any other Thelohania species represented by DNA data so far, and thus imply species from crayfish and ants may not belong in this genus. Results also confirm that genus Thelohania and family Thelohanidae are in need of revision.     

Cryptosporidium anserinum sp. n. (Sporozoa) in a Domestic Goose Anser anser L., from Iowa

SYNOPSIS. Cryptosporidium anserinum sp. n. is described in the large intestinal mucosa of a domestic goose, Anser anser L., from Iowa. Ultrastructural studies revealed schizonts containing 4 merozoites and macrogametes and schizonts with characteristic attachment zones and double membranes. The only lesion associated with infection was shortening of the intestinal microvilli.     

Functional analysis of the PsbP-like protein (sll1418) in Synechocystis sp. PCC 6803

A recent proteomic analysis of the thylakoid lumen of Arabidopsis thaliana revealed the presence of several PsbP-like proteins, and a homologue to this gene family was detected in the genome of the cyanobacterium Synechocystis sp. PCC 6803 (Schubert M, Petersson UA, Haas BJ, Funk C, Schröder WP, Kieselbach T (2002) J Biol Chem 277, 8354–8365). Using a peptide-directed antibody against this cyanobacterial PsbP-like protein (sll1418) we could show that it was localized in the thylakoid membrane and associated with Photosystem II. While salt washes did not remove the PsbP-like protein from the thylakoid membrane, it was partially lost during the detergent-based isolation of PSII membrane fractions. In total cell extracts this protein is present in the same amount as the extrinsic PsbO protein. We did not see any significant functional difference between the wild-type and a PsbP-like insertion mutant.     

Merlinius paniculoides n. sp. from Italy (Tylenchina: Belonolaimidae)

Merlinius paniculoides n. sp. is described from a bisexual Italian population found in northern Italy. This species has a coarsely annulated cuticle with approximately 30 longitudinal striae in addition to the six lateral field incisures; consequently the cuticle appears tessellate. The female tail is subcylindrical with a bluntly, rounded, non-annulated terminus. Merlinius paniculoides shares the tessellate cuticular ornamentation and the offset labial region with M. lenorus (Brown, 1956) Siddiqi, 1970 and M. tartuensis (Krall, 1959) Siddiqi, 1970. It differs from M. lenorus by the higher c and lower c'' ratios (c = 16-19 vs. 12-15 and c'' = 2.2-2,8 vs. 3.7) and greater number of longitudinal striae (28-32 vs. 24). It differs from M. tartuensis by a fewer number of annuti from stylet knob base to lip constriction (8-9 vs. 21), shorter stylet (19-21 vs. 24 μm), presence of males, and higher c'' value (c'' = 2.2-2.8 vs. 1.9).     

Ultrastructural study of the epidermis in two free-living Platyhelminthes, Mesostoma viaregginum and M. productum (Rhabdocoela, Typhloplanida)

Abstract. The epidermis of the free-living typhloplanids Mesostoma viaregginum and M. productum (Mesostominae) is described. In both species, the epidermis has polarized cells with nuclei located at the basal part of the cell, whereas mitochondria are in the apical one. The epidermis is entirely covered by microvilli and locomotory cilia anchored in the cytoplasm by vertical and horizontal rootlets. Rootlets exhibit distinct length and periodic structure in the two species. Furthermore, in each species vertical and horizontal rootlets possess different periodic structure. The pattern of termination of microtubules in epidermal cilia is described for the first time in the Typhloplanida; central microtubules shift along one axonemal side, doublets 1 and 6–9 lose their microtubule B, and gradually peripheral doublets become singlets. Finally, an electron-dense material caps the tip of the cilia. This pattern of termination closely resembles that of Temnocephalida, Kalytorhynchia, and Dalyelliida examined so far, but differences exist.     

Light and electron microscope study of Thelohania solenopsae n. sp. (Microsporida: Protozoa) in the red imported fire ant,Solenopsis invicta

A new species of Microsporida, Thelohania solenopsae, is described from the red imported fire ant, Solenopsis invicta. The Thelohania infections are localized in the fat body of workers. Meronts causing infections of progeny are found in the ovaries of queens. Spores occur only in adult ants and only vegetative stages are present in larvae and pupae. Both uninucleate octospores (eight spores within a pansporoblast membrane) and binucleate free spores (spores developing in isolation) are formed.     

Eimeria crotalviridis sp. n. from Prairie Rattlesnakes, Crotalus viridis viridis, in New Mexico with Data on Excystation of Sporozoites and Ultrastructure of the Oocyst Wall

SYNOPSIS Oocysts of Eimeria crotalviridis sp. n. are described from prairie rattlesnakes, Crotalus viridis viridis in New Mexico on the basis of light and electron microscopy and in vitro excystation of sporozoites. Sporulated oocysts of E. crotalviridis are elliptical, 26.4 × 22.3 (23–29 × 20–24) μm with ovoid sporocysts 11.7 × 8.1 (11–13 × 7–9) μm. A micropyle, micropyle cap and polar bodies are absent, but oocyst and sporocyst residua and Stieda and substieda bodies are present. Excysted sporozoites are 12.4 × 2.8 (11–13 × 2–3) μm and have 1 large posterior refractile body and a nucleus with a prominent nucleolus. Ultrastructurally, the oocyst wall has 2 layers, a thick, electron-dense, highly sculptured outer layer composed of a fine granular matrix and a thin, granular, osmiophilic inner layer, separated from the outer layer by at least one unit membrane. These layers are 441 (353–510) and 21.6 (19–29) nm thick, respectively. Within 15 min after exposure to a trypsin-sodium taurocholate fluid, sporozoites of E. crotalviridis excysted from 5-month-old sporocysts.     

Vibrio sp. as a potentially important member of the Black Band Disease (BBD) consortium in Favia sp. corals

Black Band Disease (BBD) is a well-described disease plaguing corals worldwide. It has been established that ecological and environmental stress factors contribute to the appearance and progression of the disease, believed to be caused by a diverse microbial consortium. We have identified and characterized Vibrio sp. associated with BBD in Eilat reef corals using both culture-dependent and -independent methods. Direct sampling using 16S rRNA gene clone libraries showed seasonal dynamics in the diversity of BBD-associated Vibrios . In the two sampling periods, BBD-associated Vibrio clones showed similarities to different groups: October samples were similar to known pathogens, while December samples were similar to general aquatic Vibrio sp. Cultured bacterial isolates of Vibrio sp. were highly homologous (≥99%) to previously documented BBD-associated bacteria from the Caribbean, Bahamas and Red Seas, and were similar to several known coral pathogens, such as Vibrio coralliilyticus . The proteolytic activity of Vibrio sp., as measured using casein- and azocasein-based assays, directly correlated with temperature elevation and peaked at 26–28 °C, with the microorganisms producing more proteases per bacterial cell or increasing the rate of proteolytic activity of the same proteases (potentially metalloproteases). This activity may promote coral tissue necrosis and aid in ensuing progression of the coral BBD.     

Eimeria filamentifera sp. n. from the Snapping Turtle, Chelydra serpentina (Linné), in Iowa

SYNOPSIS. Oocysts of Eimeria filamentifera sp. n. were isolated from 2 of 2 snapping turtles, Chelydra serpentina (Linné), in Iowa. The ovoid to ellipsoid oocysts of this species averaged 23.2 × 18.6 μm and had a length/width ratio ranging from 1.1 to 1.6. The ellipsoid sporocysts averaged 14.0 × 7.7 μm and had papillose, filament-bearing Stieda bodies. Oocyst and sporocyst residua were present, but micropyle and polar granule were not. This appears to be the 2nd species of Eimeria reported from snapping turtles, with E. chelydrae being the first.     

Sphaeromyxa maiyai sp. n. (Protozoa: Myxosporidea), Coelozoic Parasite of the Pacific Tomcod,Microgadus proxiimus*

SYNOPSIS. Sphaeromyxa maiyai sp. n. (Myxosporidea: Myxiidae) is described from the gall bladder of the Pacific tomcod Microgadus proximus Girard, from Newport, Oregon. The plasmodial stage is discoid averaging between 1 and 2 cm. The spore averages 27.6 × 5.6 μm in breadth, with pyriform polar capsules measuring 9.3 × 3.7 μm. Scanning electron microscopy reveals the presence of numerous longitudinal striations.     

Diplodinium (Ostracodinium) minorum sp. n., Ciliate from the Rumen of Domestic Sheep*

SYNOPSIS. Diplodinium (Ostracodinium) minorum sp. n., observed in rumen contents from several domestic sheep, resembles Diplodinium (O.) ypsilon and Diplodinium (O.) magnum. It differs from the latter 2 species primarily in the size and proportions of the body, size of the rectum and anus, and in the presence of a short right caudal lobe in many individuals. In the samples studied, D. (O.) minorum constituted from 0.06 to 0.67% of the total rumen ciliate population.