Another chloromyxid lineage: molecular phylogeny and redescription of Chloromyxum careni from the Asian horned frog Megophrys nasuta

Infection with Chloromyxum careni Mutschmann, 1999 was found in the Asian horned frog Megophrys nasuta from Malaysia and Indonesia. Kidney was the only organ infected. Coelozoic plasmodia up to 300 μm were localized in Bowman's space, embracing the glomerulus from all sides, or rarely in lumina of renal tubules. Plasmodia are polysporic, containing disporic pansporoblasts. Myxospores observed by light microscopy are colorless, variable in shape and size, measuring 6.0-8.5 × 5.0-6.5 μm, composed of two symmetrical valves joined by a meridian suture, containing four pyriform polar capsules 3.0-4.0 × 2.5-3.0 μm and a single sporoplasm. Each valve possesses 14-24 (median 21) fine longitudinal ridges clearly visible only in scanning electron microscopy. Rarely, atypical spores with a markedly pointed posterior pole and only 6-10 surface ridges are present in plasmodia together with typical spores. Both small subunit (SSU) and large subunit (LSU) rRNA gene sequences possess extremely long GU-rich inserts. In all SSU and LSU rDNA-based phylogenetic analyses, C. careni clustered as a distinct basal branch to the Myxobolus+Myxidium lieberkuehni clade, out of the marine Chloromyxum clade containing Chloromyxum leydigi, the type species of the genus. These morphological and phylogenetic data suggest erection of a new genus for the C. careni lineage, but we conservatively treat it as a Chloromyxum sensu lato until more information is available.     

Fine structure of Chloromyxum menticirrhi n. sp. (Myxozoa) infecting the urinary bladder of the marine teleost Menticirrhus americanus (Sciaenidae) in Southern Brazil

A myxosporidian was found in the urinary bladder of the teleost Menticirrhus americanus Linnaeus, 1758 (Sciaenidae) collected from the South Atlantic coast of Brazil. Polysporic amoeboid plasmodia containing sporoblasts, developing pansporoblasts and spores were free in the bladder lumen. The prevalence of infection was 17.64% (15/85). Unfixed spores were spherical to subspherical, on average 10.5 μm long, 9.8 μm wide and 10.1 μm thick (n=25), and fixed spores measured 10.1×9.5×9.7 μm. The two spore valves were of equal size and each possessed prominent sutural lines and about 41 (37–45) surface ridges aligned parallel with the suture line. These ridges gave transverse sections a cog-wheel-like outline. The spores contained four pyriform polar capsules of equal size (3.20×2.0 μm) (n=25) (fixed), each with a polar filament having 3–4 (rarely 5) coils. The binucleate sporoplasm was irregular in shape, with granular matrix and randomly distributed dense bodies. The shape and dimensions of the spore, as well as the number, position and arrangement of the surface ridges, polar capsules and polar filament indicate that this is a new species, herein designated Chloromyxum menticirrhi. The gill, liver, gall bladder and intestine of the host showed no abnormalities.     

A new myxozoan from feral goldfish (Carassius auratus)

In February 2004, a mass die-off of common goldfish Carassius auratus L., presumptively caused by bacterial coldwater disease (Flavobacterium psychrophilum), occurred at Fern Ridge Reservoir, Oregon. A range of size classes was affected, but all mature fish were female and all fish were infected with a single myxozoan, Chloromyxum auratum n. sp. No histological changes were observed associated with the parasite. Infection was represented by mictosporic plasmodia and free-floating spores in the gall bladder. Parasite spores were nearly spherical, 13.6 microm long x 12.6 microm wide x 13.1 microm thick, and possessed 4 equal-sized polar capsules. Spores had a coglike appearance in apical view because of distinct ridges 2.1 microm high protruding from the valve cells. There were 6-9 extrasutural ridges per valve (15-20 ridges per spore), aligned along the longitudinal axis, with some branching, and convergence at both poles. Morphologically, spores identified most closely with Chloromyxum cristatum Léger, 1906; however, 18S rDNA sequence data indicated only 97.5% similarity over 2,076 bp with Chloromyxum cyprini, the only synonym of C. cristatum for which DNA data are available; additional sequence data may reveal the other synonyms to be distinct species. This is the first record of a species of Chloromyxum from goldfish.     

Species of the genus Chloromyxum Mingazzini (Myxozoa: Myxosporea) infecting burbot (Lota lota L.)

Three different species of the genus Chloromyxum Mingazzini were found in burbot (Lota lota L.) collected in south-west Bohemia (Czechoslovakia). Comparison with existing records revealed that one species could be identified as C. pseudomucronatum Kashkovskiy, 1966. Found in the urinary bladder, it had subspherical spores with fine surface ridges and polysporic plasmodia. The two other myxosporea were established as new species: C. lenorae n. sp. was found in the kidney, renal corpuscles, renal tubules and interstitium, and had ellipsoid spores with surface ridges barely perceptible in the light microscope but clearly revealed by transmission electron microscopy. In the polysporic plasmodia, spores developed in pansporoblasts. C. reticulatum n. sp. was found in the gall bladder. It had polysporic plasmodia and spherical spores (average size 8.1 m in diameter) with a unique surface structures: elevated crests marking off irregular fields which appeared as a reticulum. In five of the fish infected with C. lenorae, bloodstream myxosporean stages of an extrasporogonic cycle were found. Further research is needed to determine whether they are stages of Sphaerospora cristata Shulman, 1962, a species also found in two of the burbot examined, or stages belonging to the Chloromyxum life cycle.     

Phylogenetic relationships amongst Chloromyxum Mingazzini, 1890 (Myxozoa: Myxosporea), and the description of six novel species from Australian elasmobranchs

Six novel species of Chloromyxum Mingazzini, 1890 are described using a whole evidence approach combining morphometric and molecular data, together with features of their biology. Elasmobranchs were collected in Australian waters, from the Great Barrier Reef, Queensland, off Lizard and Heron Islands; from Moreton Bay, southeast Queensland; off Hobart, Tasmania; and from the Tamar River, Launceston, Tasmania. The novel species proposed here are: Chloromyxum hemiscyllii n.sp. from Hemiscyllium ocellatum; Chloromyxum kuhlii n.sp. from Neotrygon kuhlii; Chloromyxum lesteri n.sp. from Cephaloscyllium laticeps; Chloromyxum mingazzinii n.sp. from Pristiophorus nudipinnis; Chloromyxum myliobati n.sp. from Myliobatis australis; and Chloromyxum squali n.sp. from Squalus acanthias. A seventh species from Squalus acanthias is also reported but due to limited material is not formally described. Molecular phylogenetic analyses revealed that the genus Chloromyxum is polyphyletic, and species from elasmobranchs form a well-supported sister clade, with the type species Chloromyxum leydigi, to all other congeneric species clustering within the freshwater myxosporean clade. Morphological analysis showed that elasmobranch-infecting species are predominantly pyriform shaped, have clearly thickened spore apex and possess caudal filaments, compared to other Chloromyxum species which are generally spherical or subspherical, and lack caudal filaments. These morphological and phylogenetic data provide further support for the erection of new genera, but we conservatively consider the species described in this study and other elasmobranch-infecting Chloromyxum species as Chloromyxum sensu strictu, whilst the freshwater teleost infecting and amphibian infecting species we will assign as Chloromyxum sensu lato, until more comprehensive data are available.     

The Fine Structure of Rhynchocystis pilosa (Sporozoa, Eugregarinida)

SYNOPSIS. The trophozoite of Rhynchocystis pilosa obtained from the seminal vesicles of the earthworm Lumbricus terrestris was studied by light and electron microscopy. The trophozoite's cortical organization is particularly interesting because of its unusual evaginations and associated fibrillar structures. The pellicle is formed by 2 concentric membranes elevated into 60–70 alternating primary and secondary ridges extending posteriad. Numerous long ‘hairs’ or cytopilia originate along the primary ridges and each contains a system of fibrils originating from an underlying longitudinal myoneme. Longitudinal rows of pores lie between adjacent pollicular ridges. Three systems of fibrils lie in the cortex of the trophozoite. A longitudinal myoneme consisting of 12–18 fibrils lies below each primary pellicular ridge. Circular myonemes lie below the pellicle in a parallel array along the length of the organism. Each myoneme consists of 4–8 fibrils structurally similar to those of the longitudinal myonemes. Pairs of fine filaments also lie in the inner pellicular membrane along the apex of each ridge. The trophozoite's anterior end is modified as an attachment organelle consisting of 30–35 delicate pellicular folds which originate at the base of an anterior papilla. The folds extend approximately 15 μ posteriad where they become continuous with the primary pellicular ridges. The nucleus lies in the cytoplasm near the posterior level of the attachment organelle and is surrounded by a double membrane perforated by numerous pores. The cytoplasm contains numerous small vesicles which may be found in dense aggregations. These aggregations often occur in proximity to Golgi complexes and certain membrane-bound bodies. Mitochondria are abundant in the cytoplasm as are large, ovoid paraglycogen bodies. Occasionally layers of granular membranes are arranged parallel to the surface of the paraglycogen bodies but also occur thruout the cytoplasm.     

Study of the cuspal ridges of the upper first molars in a modern Japanese population

Materials used were dental casts of the upper first molars of modern Japanese subjects, comprising 29 males and 25 females. Their molar occlusal surfaces were photographed by moiré contourography using the standard trigonal plane. The ridges of a cusp, comprising a central ridge and mesial and distal accessory ridges, were identified from the patterns of the moiré fringes. The central ridge was observed in all cusps except for the hypocone in both sexes. Frequencies of the mesial and distal accessory ridges of trigonal cusps were over 90% except for the distal accessory ridge of the metacone, and those of the hypocone were under 25% in both sexes. These values were generally higher in males than in females, especially for the distal accessory ridge of the metacone. The running pattern of the cuspal ridges showed little difference between sexes. The oblique ridge which was higher than the central groove formed a saddle-like structure. This ridge was observed in all materials, but its heights and structural components varied remarkably. In this study, the distal accessory ridge of the metacone was found to be incorporated into the oblique ridge in about 13% of cases. Variability in the running pattern of the ridges within a single cusp was highest in the hypocone and lowest in both the paracone and protocone. The results obtained are considered to represent the stability or reductive tendency of cusps in the upper first molars.     

Scanning Electron Microscope Observations on Some Life History Stages of Carchesium polypinum (Ciliata,Peritrichia)1

SYNOPSIS. Sessile zooids, and mobile telotrochs and microgamonts of Carchesium polypinum (Protozoa, Ciliata, Peritrichia), were examined by scanning electron microscopy. The results were compared to earlier light and electron microscope studies in order to investigate structural changes concerned with adaptation and differentiation. Telotrochs and microgamonts always had a contracted peristome and usually had a long phalange of cilia. Striae around the contracted buccal apparatus in all 3 stages were convoluted and often had thickened margins; those in telotrochs and microgamonts had oral-aboral ectoplasmic cross-connections. Nonbuccal striae of telotrochs and microgamonts varied in structure and height differences between epiplasmic peaks and alveoli surface membranes. The number of striae were constant in all 3 stages. Pellicular pore structure did not vary in any of the stages examined and resembled parasomal sacs located near buccal structures. Fully relaxed sessile zooids had ectoplasmic ridges coursing from polykinety kinetosomes and cilia to an area in front of the ciliated portion of the haplokinety; these ridges were interpreted to be the interkinetal fibers. Telotroch bands of sessile zooids consisted of 2 or 3 parallel ectoplasmic ridges which circled the aboral region and contained structures resembling pores. Telotroch bands in telotrochs and microgamonts had 2 enlarged, parallel ectoplasmic ridges circling the aboral region; telotroch band cilia were found between these ridges. In addition, a fold-like, ectoplasmic structure extended beyond the 2 ridges and was located between the telotroch band cilia and the aboral ridge. The epiplasmic shelf surrounding the stalk in sessile zooids was enlarged in telotrochs, and cilia were seen in the scopula depression. No scopula organelle was seen in any microgamont.     

Ceratomyxa hungarica n. sp. and Chloromyxum proterorhini n. sp. (Myxozoa: Myxosporea) from the freshwater goby Proterorhinus marmoratus (Pallas)

When studying the parasite fauna of freshwater gobies Proterorhinus marmoratus collected from the reaches of the River Danube around Budapest, two species of Myxosporea were recovered, a renal and a gall-bladder form. Previously no myxosporeans had been reported from this fish species. The spores and pseudoplasmodia of the parasite described as Ceratomyxa hungarica n. sp. were found in the convoluted tubules of the kidney and in the cavity of Bowman's capsule. The pseudoplasmodia were loosely attached to the wall of the tubules, causing their distention. Within each pseudoplasmodium two spores were formed. In the case of Chloromyxum proterorhini n. sp. only spores floating freely in the contents of the gall-bladder were found. Since Ceratomyxa species are typically marine fish parasites, Proterorhinus marmoratus, a fish species which has adapted to freshwater, appears to have retained some of its marine myxosporean fauna.     

Sphaerospora ovophila N. Sp. and Myxobolus algonquinensis N. Sp. (Myxozoa, Myxosporea), Ovarian Parasites of Fish from Algonquin Park, Ontario, Canada

ABSTRACT. Sphaerospora ovophila n. sp. and Myxobolus algonquinensis n. sp., found in the ovary of the pumpkinseed ( Lepomis gibbosus ) and the golden shiner ( Noremigonus crysoleucas ) respectively from Algonquin Park, Ontario, are described using light and electron microscopy. Ovoid cysts of S. ovophila measured up to 500 pm in length. Monosporic pseudoplasmodia were ovoid or ellipsoid in shape and measured up to 12.5 pm in length. Spores were 7.2–8.4 pm long × 6.0–7.0 pm wide (in sutural plane) × 7.4–8.2 pm thick (perpendicular to sutural plane), with two subspherical polar capsules of equal size measuring 2.7–3.2 × 2.6–3.1 pm and each of which contained a polar filament with 6–7 coils. The spore had a straight sutural ridge which protruded slightly at the anterior end and contained two uninucleate sporoplasms. The spore valve had ornate folds on the posterior end. Cysts of M. algonquinensis ranged from ovoid to elongated ellipsoid in shape and measured up to 800 pm in length. Mature spores measured 13.G15.7 pm long × 10.1–12.1 pm wide × 5.0–6.9 pm thick. with two pyriform polar capsules of equal size measuring 5.1–5.5 pm × 2.5–2.9 pm, each of which contained a polar filament with 4–6 coils. The spores of M. algonquinensis had smooth valves, a straight sutural ridge and a distinct small intercapsular appendix.     

Dictyostelium discoideum fruiting bodies observed by scanning electron microscopy.

The fruiting bodies of Dictyostelium discoideum, fixed in a vapor of 25% glutaraldehyde, were observed by scanning electron microscopy. The surface of the basal disk had rootlike ripples, sometimes with concentric rings of riblike striations at its periphery. The surface of the stalk had pleated and twisted ridges along its entire length, but no lateral projections. The spores were randomly orientated and had a crinkled surface. The surface of the lower part of the papilla had flattened ridges running horizontally. A slime coat remained in the region of transition between the stalk and sporangium, but not on the spores.     

Myxosoma funduli Kudo (Myxosporida) in Fundulus kansae: Ultrastructure of the Plasmodium Wall and of Sporogenesis*

SYNOPSIS Ultrastructure of the plasmodium wall and of sporogenesis were studied in Myxosoma funduli Kudo infecting the gills of Fundulus kansae (Garman). Plasmodia were located within the lamellar tissues adjacent to sinuses and capillaries. The plasmodium wall consisted of a single unit membrane which was continuous with numerous pinocytic canals extending into the parasite ectoplasm. The plasmodium membrane was covered by a surface coat of almost uniform thickness which prevented direct parasite-host cell contact. Numerous generative cells and cell aggregates, representing early stages of spore development, were seen in immature plasmodia. Later stages of spore development, including mature spores, were observed in older plasmodia. Sporogenesis was initiated by envelopment of one generative cell, the sporont, by a 2nd, nondividing cell, the envelope cell. The sporont and its progeny proceeded through a series of divisions until there were 10 cells, all compartmentalized within the envelope cell. Subsequently, the 10 cells became structurally differentiated and arranged into two 5-celled spore-producing units, each consisting of 1 binucleate sporoplasm and 2 capsulogenic cells, all surrounded by 2 valvogenic cells. Observations of later developmental stages revealed the major events of capsulogenesis, valvogenesis, and sporoplasm maturation, which occurred concomitantly during spore construction.     

Life Cycle of Culicospora magna (Kudo, 1920) (Microsporida: Culicosporidae) in Culex restuans Theobald with Special Reference to Sexuality1

The life cycle of Culicospora magna (Kudo, 1920) Weiser, 1977, consists of two major developmental sequences that alternate in host individuals of successive generations, each of the sequences starting with a sporoplasm and ending with spores. The first sequence occurs in larval, pupal, and adult stages of a parental generation of the host mosquito, Culex restuans Theobald; it begins with a sporoplasm from an ingested uninucleate spore and progresses through stages in gametogony, plasmogamy, nuclear association, merogony, karyogamy, and disporous sporulation with production of binucleate spores that discharge sporoplasms into the oocytes. The second sequence occurs in egg and larval stages of a filial generation of the same host species; it begins with the binucleate sporoplasm that entered the egg, includes stages in merogony, nuclear dissociation, and mictosporous sporulation, and ends with uninucleate spores. These spores are released into the environment following death of the host and are capable of infecting new parental generation host individuals. The life cycle is conceived as an alternation of generations related to haploidy and diploidy in the nuclei, the transition from haploidy to diploidy occurring with nuclear association and the transition from diploidy to haploidy occurring with nuclear dissociation.     

Moth olfactory trichoid sensilla exhibit nanoscale-level heterogeneity in surface lipid properties

Chemical force microscopy (CFM) based on tapping mode Atomic force microscopy (AFM) utilized with topographic and phase-shift analyses was used to investigate the topography and surface chemical properties, respectively, of the long trichoid sensilla on the antennae of male Helicoverpa zea. AFM topographic imaging revealed regular series of step-ridges along nearly the entire length of each sensillum, except for the basal ca. 1/3 portions, which were devoid of such ridges. Inter-ridge regions were flat, with regularly spaced pores, ca. 30 nm in diameter populating these planar areas. Many pores exhibited a raised dome that often nearly completely spanned the depression, with only the edges of the depressed portion of the pore still visible. Some pores were observed also along the bases of the ridges. CFM probing of the surface for chemical interactions with the SiO₂ hydrophilic tip revealed consistently diminished hydrogen bonding of the ridge edge areas with the tip than along the flat planar inter-ridge regions. Surfaces of domes over the pores also tended to have less hydrogen bonding with the tip than the planar surfaces. Functionalizing the CFM tip by bonding octadecyl-hydrocarbon to it eliminated these surface chemical-CFM tip interactions and no differences in tip interaction with the sensillar surfaces were observed. Trichoid sensilla from the male antennae of a second species, Utethesia ornatrix, did not exhibit similar heterogeneity between ridge edges versus planar areas with regard to hydrogen bonding with the SiO₂ hydrophilic tip. Pores on U. ornatrix sensilla occurred only along the bases of ridges on their trichoid sensilla. We suggest that the surface lipids of the H. zea sensilla are distributed in a chemically heterogeneous fashion to aid adsorption and transport of aldehyde pheromone component molecules through the pores into the sensillum lumen, possibly through solubilization in an epicuticular lipid layer. The trichoid sensilla of U. ornatrix do not exhibit such surface chemical heterogeneity, and this species-difference may be due to the usage by U. ornatrix of hydrocarbon molecules rather than aldehydes for their sex pheromone components.     

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.     

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.     

东北薄荷的化学型

东北薄荷的化学型桂新,周自新（安徽中医学院中药系合肥２３００３８）（南京市卫生防疫站南京２１０００３）周荣汉（中国药科大学植物化学分类研究室南京２１００３８）ＣｈｅｍｏｔｙｐｅｓｏｆＭｅｎｔｈａｓａｃｈａｌｉｎｅｎｓｉｓＫｕｄｏ￥Ｃｈｏｕ－Ｘｉｎ（Ａ...     

Studies on the Pollen Morphology of Chinese Ephedra

Pollen morphology of 11 species of Chinese Ephedra has been studied All of them were examined under the light microscope and scanning electron microscope. Pollen grains are long ellipsoidal or ellipsoidal in shape. The long axis is 40 to 67 μm . and the short one is 21 to 38 pm . No aperture is present. The ridges vary from 5 to 16. The width of the ridge is 2.6–6.1 μm , Hyaline line is distinct or indistinct, or absent, and in some species it branches. According to the type of pollen grains of Ephedra divided by steeves and Barghoorn (1959), it is easy to separate type A from type D, but it is difficult to distinguish type B from type C. Thus we combine type B with type C into one type (type BC). E. intermedia, E. siniea, E. equisetina belong to type A, and E. likiangensis, E. likiangensis f. mairei and E. minute to type D. All other species belong to type BC. Type A plants distribute in Northwest, Northeast, and North China, type D in Southwest China and Tibet, and type BC is found almost over China except in the regions of lower valley of Yangtze River and in the valley of the Pearl River. It is noted that the pollen grains of Ephedra and spores of Schizeae are easily confused sometimes in pollen analysis. When we compare their morphology, two main different po- ints are found: 1. The end of pollen grain of Ephidra is sharper, but that of the spore of Schizeae is more flat. 2. The ridges of pollen grain of Ephedra come nearer each other towards the ends, and the ribs of spores of Schizeae are parallel through the ends.