Alternation of Actinosporean and Myxosporean Phases in the Life Cycle of Zschokkella nova (Myxozoa)

ABSTRACT. Experimental evidence has been gathered to show that the life cycle of the myxozoan gallbladder parasite Zschokkella nova Klokacewa, 1914, which infects the fish Carassius carassius , has a complex life cycle with alternation of two hosts (fish and Oligochaeta) and two developmental phases (myxosporean and actinosporean). The gut epithelium of the oligochaete, Tubifex tubifex , exposed experimentally to Z. nova , obtained from C. carassius , became infected with organisms resembling Actinosporea. The spore structure and cube-like network of the interconnected spores is reminiscent of Siedleckiella silesica Janiszewska, 1952, although the spores are very different in size and number of sporoplasm nuclei. The life cycle of Z. nova resembles that of the whirling disease agent Myxosoma cerebralis described by Wolf and Markiw, which also alternates between fish and oligochaete hosts.     

Molecular relationships and phylogeny in a community of myxosporeans and actinosporeans based on their 18S rDNA sequences

The community of myxosporeans and actinosporeans inhabiting a typical Scottish highland stream and the outflow area of an adjacent salmon hatchery was analysed on the basis of their 18S rDNA sequences. Nine myxosporeans belonging to the genera Sphaerospora, Chloromyxum, Zschokkella, Myxidium, Hoferellus and Myxobilatus were identified from mature spores in different organs of the fish species present. Twelve actinosporean types belonging to the collective groups of neoactinomyxum, aurantiactinomyxon, raabeia, echinactinomyxon and synactinomyxon were found to be released from oligochaete worms collected from sediments. Twenty of the 21 sequences obtained from these myxozoans are new entries to the myxozoan database, and the genera Chloromyxum, Hoferellus and Myxobilatus were entered for the first time. Study of the molecular relationships between the different taxa and with other myxozoan sequences available showed that the myxosporeans inhabiting the urinary system clearly cluster together, independently of host species or spore morphology. However, the sequences of the two Sphaerospora species encountered show considerable differences from other members of this group and all other freshwater myxosporeans, and they were found to occupy an ancestral marine position. Three actinosporeans, i.e. Neoactinomyxum eiseniellae, Aurantiactinomyxon pavinsis and Raabeia 'type 3' were found to represent alternate life cycle stages of Chloromyxum sp., Chloromyxum truttae and Myxidium truttae, respectively (approximately 1400 identical base pairs each). Three other actinosporeans encountered (two echinactinomyxon and one raabeia type) showed over 92% sequence identity with myxosporeans from GenBank, whereas all other actinosporeans formed a closely related group devoid of any known myxosporeans.     

Trypanosoma avium: Fine Structure of All Developmental Stages*

SYNOPSIS. Thin sections of the following stages of Trypanosoma avium were examined in the electron microscope: Trypomastigote forms from the blood of a bird, large epimastigote forms developing from the former after 2 hours in vitro, small epimastigote and metacyclic trypomastigote forms developing after longer periods of cultivation in vitro. The general structure of all stages was similar to that which is already well known for the genus, with the following points being of particular interest: (1) In the large trypomastigote and epimastigote forms, and possibly also in the smaller forms, the flagellar sheath was attached to the pellicle, at least in places. In the large trypomastigote forms, this resulted in the drawing out of a “fin” or ridge of cytoplasm, particularly in the mid-region of the body, to form a true undulating membrane. (2) At least some of the individuals in the blood of a bird have 2 basal bodies, one of which is aflagellate, altho these individuals rarely if ever divide. The large epimastigote forms into which they transform in vitro develop 4 basal bodies (2 flagellate and 2 aflagellate) before dividing. (3) The chondriome is well-developed in all stages, extending thruout the body, even to the tip of the elongated posterior end of the form in the avian host. (4) A short cytostome, leading from the flagellar pocket, was seen in the hematozoic (blood-inhabiting) trypomastigote form but not in other stages. (5) It is suggested that the forward movement of the kinetoplast and basal body during the transformation from trypomastigote to epimastigote form is mediated by localized cytoplasmic movement, resulting in the “rolling-up” of the organism's hind end. It is further suggested that protein synthesis is reduced or even suppressed entirely in the small epi- and trypomastigote forms appearing at the end of the developmental cycle in vitro or in the insect host, such synthesis recommencing rapidly after re-entry into the vertebrate.     

Fine Structure of the Sporogonic Stages of Nosema parkeri

SYNOPSIS The fine structure of sporogonic stages of Nosema parkeri Krinsky, a microsporidan from the argasid tick, Ornithodoros parkeri Cooley, is described. Developmental changes in the spore coat and cytoplasmic organelles are discussed. As a sporoblast transforms into a spore, the organelles become more compact and the membranes surrounding them appear to become more taut. It is suggested that the polaroplast complex is involved in fluid transport during development of the spore. Organelles in the mature spore include 2 contiguous nuclei enveloped in a lattice containing ribosome-like particles, a polaroplast complex composed of laminar and saccular regions, and a coiled tubular polar filament attached to a polar sac. Sporogonic stages do not appear to have mitochondria, Golgi apparatus, or a posterior vacuole. The fine structure of the spore of N. parkeri is very similar to that of species of Nosema found in insects, crustaceans, and trematodes.     

Fine Structure of Cephalic Sense Organs in Heterodera glycines Males

Cephalic sense organs of Heterodera glycines males were examined in detail by electron microscopy. Each amphid basically consists of an amphidial gland, a nerve bundle, and an amphidial duct. The amphidial gland consists of a microvillous region, and laterally is closely associated with a large secretory cell. The nerve bundle penetrates the microvillous region, and further anteriorly individual nerve processes (dendrites) separate from one another, thus forming a sensilla pouch which is enveloped by the microvillous region of the gland. Anterior to the pouch, the cilia-like dendrites converge as they enter and eventually terminate in the amphidial duct. Heterodera glvcines males have the innervation basis for a full complement of sixteen papillae, although surface manifestations are present for only six minute inner labial papillae. In addition, four outer labial and four cephalic receptors terminate beneath the surface, and another two dendrite pairs end further posteriorly beneath the basal plate of the cephalic framework. Papillary receptors which terminate beneath the surface are probably mechanoreceptive, whereas inner labial papillae have pore-like openings to the exterior and may be chemoreceptive. Amphids and papillae of H. glycines are fundamentally similar to those of Meloidogyne incognita, although certain striking differences exist.     

Fine Structure of Fossilized Bacteria in Volyn Kerite

Ultrathin sectioning and cryofracture of fibrouskerite, sampled from 1.8–1.75 billion year old Volynsediments (Ukraine), revealed in bacteria-like bodies thepresence of structures similar to sheath, cell wall,periplasm, cytoplasm, septum, membranes, intramembraneparticles, poly--hydroxybutyrate inclusions. On thestrength of these data and also the fatty acid profilesof these microfossils, we concluded that fibrous keritesare biogenic formations, namely fossilized bacterial mats.     

Gametogenesis and Sporogony of Hepatozoon Mocassini (Apicomplexa: Adeleina: Hepatozoidae) In an Experimental Mosquito Host, Aedes Aegypti

ABSTRACT. The sexual life cycle of the hemogregarine Hepatozoon mocassini was studied in Aedes aegypti , an experimental mosquito host, using transmission electron microscopy. Gamonts were observed leaving the host snake erythrocyte as early as 30 min after mosquitoes ingested infected blood, and some gamonts had penetrated the gut epithelial cells by this time. Six hours post-feeding, gamonts were identified within cells of the abdominal fat body. Twenty-four hours post-feeding, gamonts were often entrapped within the peritrophic membrane, but were no longer observed within the gut wall. Parasites pairing up in syzygy and undergoing sexual differentiatioe were observed within fat cells at this time, and by 48 hours post-feeding, well-developed macro- and microgametocytes as well as microgametes were discernible. Developing zygotes observed 3 days post-feeding were enclosed within a panoitophorous vacuole. By day 6, multinucleate oocysts with crystalloid bodies in the cytoplasm were seen. Sporazoites developing within sporocysts appeared by day 12. Seventeen days post-feeding, mature oocysts with sporocysts containing approximately 16 sporozoites were observed upon dissection of mosquitoes. Large crystalloid bodies no longer bound by rough endopbsmic reticulum were located anterior and posterior to the sporozoite nucleus. Free sporozoites were not observed.     

Fine Chloroplast Structure in Cucumber and Pea Leaves Developed under Red Light

Photosynthetic activity, the content of various photosynthetic pigments, and the chloroplast ultrastructure were examined in the leaves of cucumber (Cucumis sativus L.) and pea (Pisum sativum L.) plants of different ages grown under red light (600–700 nm, 100 W/m²). In pea leaves tolerant to red-light irradiation, chloroplast ultrastructure did not essentially change. In the first true leaves of cucumber plants susceptible to red-light irradiation, we observed a considerable increase in the number and size of plastoglobules, the appearance of chloroplasts lacking grana or containing only infrequent grana, and stromal thylakoids. In the upper leaves of 22-day-old cucumber plants, the chloroplast structure was essentially similar to that of the control chloroplasts in white light, and we therefore suppose that these plants have acclimated to red light.     

Fine Structure of a New Human Microsporidian, Encephalitozoon hellem, in Culture

Encephalitozoon hellem is a new human microsporidian isolated from corneal biopsies and conjunctival scrapings of three AIDS patients and cultured in Madin Darby canine kidney (MDCK) cells. Encephalitozoon hellem and Encephalitozoon cuniculi display different protein profiles with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and unique antibody binding patterns with murine antisera against Western blots of each organism. Developmental stages of E. hellem in culture are similar to E. cuniculi. Meronts are 1.3–2.7 μm in diameter, develop within a parasitophorous vacuole adjacent to the vacuolar membrane, divide by binary fission, and contain one or two discrete nuclei. Sporonts measure 2 × 3 μm, separate from the vacuolar membrane, and have a thickened outer membrane. Sporoblasts display a tri-layered wall and possess the earliest recognized polar filaments. Mature spores measure 1 × 1.5 μm and are more electron-dense than other stages. Each spore contains a single nucleus, a polar tubule with four to nine coils, thin electron-dense exospore and thick, electron-lucent endospore. Although E. hellem and E. cuniculi differ biochemically and immunologically, their fine structure and development are indistinguishable.     

Fine Structure of Cephalic Sense Organs in Meloidogyne incognita Males

Amphids, and the cephalic and labial papillae of Meloidogyne incognita males were examined in detail by electron microscopy. Each amphid basically consists of an amphidial gland, a nerve bundle and an amphidial duct. The gland is a broad microvillous organ with a narrow anterior process, which is closely associated with the amphidial duct. A posterior process of the gland contains secretory organelles and proceeds along the esophagus with the lateral cephalic nerve bundle. The nerve bundle penetrates the broad portion of the gland and, subsequently, individual nerve processes (dendrites) separate from one another, thus forming the sensilla pouch which is enveloped by the gland. Anterior to the pouch, the dendrites converge as they enter and eventually terminate in the amphidial duct. The external opening of the duct is a broad slit which separates the cheek, the outermost part of the lateral lip, from the remainder of the lip region. M. incognita males have six inner labial papillae and four outer cephalic papillae which are each innervated by two and one cilia, respectively. In labial papillae, the cilia appear to terminate at the base of a pore opening, whereas in cephalic papillae each cilium terminates beneath the labial cuticle.     

A Fine Structure Study of Physarum polycephalum During Transformation from Sclerotium to Plasmodium: A Six-Stage Description

ABSTRACT. Physarum polycephalum is classified presently as a sarcodinid in the class Eumycetozoea. It produces a sclerotial dormant stage consisting of a crustose deposit containing nucleated spherules of cytoplasm enclosed within a honey-comb-like matrix of organic walls. When rehydrated, the sclerotium reverts to a plasmodium: 1) the spherules become increasingly vacuolated, 2) electron-dense granules become dispersed within expanding vacuoles, and 3) pseudopodial extensions develop from the periphery of the spherule cytoplasm, penetrating the fragmenting walls, and making interconnections with surrounding spherules, eventually leading to a fully reticulated plasmodium. Six stages are identified during reversion from sclerotium to plasmodium in laboratory cultures, and their successive appearance was mapped over time. The six stages are: 1) sclerotial stage with crenulated nuclei, 2) cytoplasmic activation with smooth nuclear envelopes, 3) initiation of pseudopodial protrusions, 4) pseudopodial penetration into or across walls, 5) cytoplasmic interconnections among spherules with wall disintegration, and 6) fully formed cytoplasmic network as plasmodium. Cytochrome c oxidase activity, expressed per unit protein content of the homogenate, remains fairly constant throughout the developmental sequence, whereas acid phosphatase activity, expressed per unit protein concentration, is somewhat lower in the sclerotium than in subsequent stages of development after hydration.     

Fine Structure of the Head and Cervical Region of Ceramonema carinatum (Chromadorida: Ceramonematidae)

Structure of the head and cervical region of Ceramonema carinatum (Chromadorida: Ceramonematidae) was described from transmission electron microscopy of serial transverse and longitudinal sections of two females. An unbroken massive cortical layer encompasses the head, except where three thin liplets surround the mouth. A large flask-shaped buccal cavity, with simpler less dense cuticle identical with that of the pharynx, abuts the body cuticle just within the mouth. Myoepithelial ceils constituting the buccal and pharyngeal regions were described. Sixteen head sensilla, the amphids, and dorsal and ventral internal sensilla were identified and described, each with associated sheath and socket cells. Ultrasturcture of the head was compared with that of other nematodes. Arrangement of sensilla resembled that of Monhysterida and Rhabditida with some significant variations, such as prominent longitudinally arranged intracellular organelles containing many microtubules associated with the amphids. The buccal cavity was almost entirely pharyngeal in character. A well-developed system of structural fibrils and abundant hemidesmosomes were notable features.     

The Early Stages of Speciation in Amazonian Forest Frogs: Phenotypic Conservatism Despite Strong Genetic Structure

Phylogeographic perspectives incorporating multiple classes of characters, especially those relating to sexual signals, are promising for the elucidation of recent evolutionary mechanisms driving speciation. Here, forest frogs were used as a model system to access distinct stages in the process of evolutionary differentiation. We studied 280 individuals assigned to three species: Allobates paleovarzensis, A. nidicola and A. masniger. Samples were collected at 20 localities arranged in two study systems, along the middle Amazon and the lower Madeira Rivers, in Central Amazonia. Mantel tests, analyses of molecular variance, and the spatial distribution of haplogroups indicated that the distribution of genetic variability, as inferred from a mitochondrial DNA marker, was determined by a combination of isolation-by-distance effects and the transposition of large Amazonian rivers. These two factors had contrasting relative influences in each of the study systems, which also differed regarding the estimated time of the major cladogenetic events. Pronounced population genetic structure was observed. However, multivariate discriminant function analyses revealed that the phenotypic (morphological and acoustic) divergence was loosely related with genetic differentiation and did not successfully predict assignment of individuals to genetic groups. The observed distribution of genetic variability showed the important role of genetic drift in the diversification of the mitochondrial marker studied. The phenotypic conservatism among populations was surprising in view of the high genetic structuring observed, and indicates a prevailing role of stabilizing selective forces in the process of sexual signal and morphological differentiation.     

Fine Structure of the Endogenous Stages of Eimeria labbeana. I. The First Generation Merozoites

SYNOPSIS. The fine structure of the 1st generation merozoites of Eimeria labbeana from the ileal mucosa of artificially infected pigeons ( Columba livia ) was investigated and described. The 1st generation merozoites which appeared between 36-48 hr after infection averaged 4.4 × 2.1 μm in size. The 3-membraned pellicle was irregular in texture and harbored a single micropore, and many micropore-like invaginations. Closely apposed to the inner pellicular membrane were seen 22 microtubules, each 22–25 nm in diameter. An apical vesicle, 50 nm in diameter, seen at the anterior extremity, was connected with the common duct of the micronemes. The conoid consisted of 9 spiral elements, each 30 × 25 nm. The paired organelle (rhoptries) varied in length (1.4–2.2 μm), and the ductules (23 nm diameter) were composed of 2 inner tubules, each 6 nm in diameter. A unit membrane enveloped the partially alveolar and differentially osmiophilic interior of the bulbous regions of the rhoptries. The "rod-like structure"was found to be tubular and represented the common duct of the micronemes.     

Fine Structure of Mummified Cells of Microorganisms Formed under the Influence of a Chemical Analogue of the Anabiosis Autoinducer

Under the influence of alkyl hydroxybenzene (C₆-AHB) added to cell suspensions at concentrations of (1–5) × 10^–3M, the cells of Saccharomyces cerevisiae, Micrococcus luteus, and Thioalkalivibrio versutusunderwent dramatic changes in the ultrastructural organization of cell membranes, cytoplasm, and inclusions. In yeast suspension, the first changes were observed after 15 min in the structure of pocket-like invaginations in the cytoplasmic membrane (CM): they were shortened and thickened. In the subsequent 30 to 60 min, CM ruptures were formed in the regions devoid of intramembrane protein particles and in the pocket-like invaginations. After 24 h, complete disintegration of the intracellular membrane structures and conglomeration of the ribosomal part of the cytoplasm occurred. Similar changes were observed on the exposure of gram-positive and gram-negative bacteria to AHB. However, the cell wall in all the microorganisms studied was not destroyed, and in Micrococcus luteusit was even thickened. These mummified forms were preserved as morphologically intact but nonviable cells for more than three years of observations. By their ultrastructural characteristics, these mummified forms of microorganisms were similar to the fossilized microorganisms discovered by us in fibrous kerite. The concept of micromummies was formulated. AHB are supposed to play an important role in the process of fossilization of microorganisms in nature.     

On the Cytology and Fine Structure of the Neogregarine Syncystis aeshnae Tuzet and Manier, 1953 (Apicomplexa, Syncystidae)

The neogregarine Syncystis aeshnae Tuzet and Manier, 1953, was obtained from two species of dragonfly larvae, Aeshna grandis and Libellula quadrimaculata , collected in southern Sweden. Merogony, gamogony, and sporogony were observed in the adipose tissue, which was destroyed, and the haemocoele. Merozoites had a traditional apical complex. Gamonts associated in pairs did not unite conoid-to-conoid, and the border between gamonts was not found to break down. Each association produced 32 spores in bilobed gametocysts. Spores were elongate, oval, 4.0–5.8 × 8.0–10.0 μm in fixed and stained condition, each with 4+4 terminal and four equatorial thread-like projections. The thick-walled spore is considered the sporocyst. Species identification, ultrastructural cytology, and problems associated with using the terms oocyst and sporocyst are discussed.     

Fine Structure of the Neogregarine Farinocystis tribolii Weiser, 1953. Developmental Stages in Merogony.

SYNOPSIS. The fine structure of schizonts and free merozoites of the neogregarine Farinocystis tribolii Weiser, and their development in the fat body of larval Tribolium castaneum were studied.
The surface of a multinucleate schizont and that of a uninucleate merozoite is covered by a double-layered membrane. Rhoptries and micronemes are present. The cytoplasm is packed with ribosomes and also contains dark bodies. Mitochondria are of the vesicular type. The spherical nucleus of the schizont and merozoite contains a large nucleolus. The anterior end of the merozoite has a typical conoidal complex composed of a conoid and a polar ring with 22 subpellicular mirotubules projecting from it.
New findings are a membranous septum across the body of the merozoite at 2/3 of its length below the nucleus and a highly osmiophilic spiral structure in the perinuclear space close to the Golgi complex. In addition, we found some "developmental stages" of the latter structure.     

Fine structure of the epistome in Phoronis ovalis: significance for the coelomic organization in Phoronida

Abstract. The hypothesis of a common ancestry of the lophophorate taxa Brachiopoda, Bryozoa, Phoronida, and the Deuterostomia can be traced back to the late 19th century when Masterman recognized a tripartite organization of the body consisting of pro-, meso-, and metasome, along with coelomic body cavities in each compartment, as characteristic for Echinodermata, Pterobranchia, Phoronida, and Brachiopoda. This idea became quite popular under the name archicoelomate concept. The organization of the phoronids, and especially of their transparent actinotroch larva, has for a long time been used as a touchstone for the validity of this concept. As a coelomic lining can reliably be recognized only on the ultrastructural level, this technique has been applied for adults of Phoronis ovalis , which is assumed to be a sister species to all other phoronids. Phoronis ovalis contains only two coelomic compartments, a posterior coelom inside the trunk (metasoma), occupying the space between the trunk epidermis and the digestive epithelium, and an anterior lophophoral coelom inside and basal to the tentacular crown (mesosoma). There is no coelomic cavity inside the epistome (prosoma). This part of the body is filled with myoepithelial cells, which are continuous with the epithelial lining of the lophophore cavity. These cells form a lumenless bilayer and possess long, tiny myofilamentous processes, which are completely embedded in an extracellular matrix. A comparison with data on P. muelleri shows that there is no need to assume three different coelomic cavities in Phoronida, in contrast to the predictions of the archicoelomate concept. At least for this taxon, a correspondence to the situation in deuterostomes can hardly be found.     

The Fine Structure of Saprodinium dentatum Lauterborn, 1908 as a Representative of the Odontostomatida (Ciliophora)

The fine structure of the sapropelic ciliate Saprodinium dentatum is described based on phase-contrast microscopy, silver-staining techniques, cryo-fracture scanning electron microscopy, and thin sections. The study concentrates on a detailed analysis of the somatic cortex and the oral ciliature of this highly asymmetric, laterally compressed ciliate. The cell shape is dominated by a number of site-specific spines and the curving course of 10 somatic kineties (SK 1–10). The SK, composed of dikinetids, show an intrakinety differentiation that seems characteristic for other odontostomes as well. The anterior segment of the SK is mostly ciliated, followed by a non-ciliated segment in which the kinetosomes lack all typical fiber systems. Except for SK 4–6, the posterior segment is ciliated again, forming the spine kinetics associated with particular caudal spines. The anterior segment of SK 3 through SK 7 form the frontal band, which together with the two frontal kineties constitutes the main locomotory organelle for a ciliate that creeps on the substratum. A short kinety with inverse polarity, not seen in earlier light microscopical studies, was observed near the oral spine. We made particular effort to find a logical explanation for the observed association of the SK with the various caudal spines. The oral ciliature consists of nine adoral organelles located in a tripartite oral cavity. The absence of a paroral ciliature together with the position of the cytostome anterior to the adoral organelles may be the result of rotational movement of the oral apparatus during the evolution of these bizarre ciliates. Results are discussed with special reference to the phylogenetic relationship of the Odontostomatida to the Heterotrichida and no conclusive answer was found in this first electron microscopical study of an odontostomatid ciliate.