Fine Structure and Development of Pilosporella chapmani (Microspora: Thelohaniidae) in the Mosquito,Aedes triseriatus (Say)1

ABSTRACT. New information on the life cycle and fine structure of Pilosporella chapmani, a microsporidium of the mosquito Aedes triseriatus, is presented. Pilosporella chapmani is shown to have two sporulation sequences, one of them being involved in transovarial transmission. One sequence, involving meiosis and production of a moniliform sporogonial plasmodium, occurs in the larval fat body, resulting in eight uninucleate, spherical, and fully developed spores. The other occurs in oenocytes of adult mosquitoes and results in isolated, binucleate, elongate, and thin-walled spores. Also, for the first time, metabolic products are shown to be expelled into the surrounding host tissues through the wall of the sporocyst.     

Life cycle,ultrastructure and molecular phylogeny of Hyalinocysta chapmani (Microsporidia: Thelohaniidae), a parasite of Culiseta melanura (Diptera: Culicidae) and Orthocyclops modestus (Copepoda: Cyclopidae)

The complete life cycle of the microsporidium Hyalinocysta chapmani is described from the primary mosquito host Culiseta melanura and the intermediate copepod host Orthocyclops modestus. Infections are initiated in larval C. melanura following the oral ingestion of uninucleate spores from infected copepods. Spores germinate within the lumen of the midgut and directly invade fat body tissue where all development occurs. Uninucleated schizonts undergo binary division (schizogony) followed by karyokinesis (nuclear division) to form diplokaryotic meronts. Merogony is by synchronous binary division. The onset of sporogony is characterized by the simultaneous secretion of a sporophorous vesicle and meiotic division of the diplokaryon resulting in the formation of eight ovoid meiospores enclosed within a sporophorous vesicle. Most infected larvae die during the fourth stadium and there is no evidence of a developmental sequence leading to vertical transmission. Hyalinocysta chapmani is horizontally transmitted to O. modestus via oral ingestion of meiospores. Infections become established within ovarian tissue of females and all parasite development is haplophasic. Uninucleate schizonts divide by binary division during an initial schizogonic cycle. Newly formed uninucleate cells produce a thin sporophorous vesicle and undergo repeated nuclear division during sporogony to produce a rosette-shaped, multinucleated sporogonial plasmodium with up to 18 nuclei. This is followed by cytoplasmic cleavage, sporogenesis, and disintegration of the sporophorous vesicle to form membrane-free uninucleate spores. Infected females eventually die and there is no egg development. The small subunit rDNA sequence of H. chapmani isolated from meiospores from C. melanura was identical to the small subunit rDNA sequence obtained from spores from O. modestus, corroborating the laboratory transmission studies and confirming the intermediary role of O. modestus in the life cycle. Phylogenetic analysis was conducted with closely related microsporidia from mosquitoes. Hyalinocysta chapmani did not cluster within described Amblyospora species and can be considered a sister group, warranting separate genus status.     

Epizootiology of Hyalinocysta chapmani (Microsporidia: Thelohaniidae) infections in field populations of Culiseta melanura (Diptera: Culicidae) and Orthocyclops modestus (Copepoda: Cyclopidae): a three-year investigation

The epizootiology, transmission dynamics and survival strategies employed by the microsporidium Hyalinocysta chapmani were examined in field populations of its primary mosquito host, Culiseta melanura and its intermediate copepod host, Orthocyclops modestus over a three-year period in an aquatic subterranean habitat. H. chapmani was enzootic and was maintained in a continuous cycle of horizontal transmission between each host. There were three distinct periods during the summer and fall when developing mosquito larvae acquired infections; each was preceded by or coincident with the detection of infected copepods. Results were corroborated in laboratory bioassays, wherein transmission was achieved in mosquito larvae that were reared in water and sediment samples taken from the site during the same time periods. The highest infection rates, ranging from 60% to 48%, were repeatedly observed during the first six weeks of larval development. These were coincident with the most sustained collections of infected copepods obtained during the year and highest levels of infection achieved in the laboratory transmission studies. The high prevalence rates of lethal infection observed in larval populations of C. melanura at this site are among the highest recorded for any mosquito-parasitic microsporidium and clearly suggest that H. chapmani is an important natural enemy of C. melanura. H. chapmani appears to overwinter in diapausing mosquito larvae but may also persist in copepods. The absence of vertical transmission in the life cycle of H. chapmani and the sole reliance on horizontal transmission via an intermediate host are unique survival strategies not seen among other mosquito-parasitic microsporidia. The epizootiological data suggest that this transmission strategy is a function of the biological attributes of the hosts and the comparatively stable environment in which they inhabit. The subterranean habitat is inundated with water throughout the year; copepods are omnipresent and C. melanura has overlapping broods. The spatial and temporal overlap of both hosts affords abundant opportunity for continuous horizontal transmission and increases the likelihood that H. chapmani will find a target host. It is hypothesized that natural selection has favored the production of meiospores in female host mosquitoes rather than congenital transfer of infection to progeny via ovarian infection as a strategy for achieving greater transmission success.     

A new cave-inhabiting Ptomaphaginus beetle from Sarawak (Leiodidae: Cholevinae)

Abstract. A new troglophilic species, Ptomaphaginus chapmani , is described from caves in Sarawak. it has the very unusual combination of very reduced eyes but functiobnal wings and may be interpreted as a phylogenetic relict, with characters suggesting a New World origin for this Oriental genus.     

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.     

AUTOINHIBITION OF SPORE GERMINATION IN NOSTOC SPONGIAEFORME (CYANOPHYCEAE)1

Medium in which cultures of Nostoc spongiaeforme Ag. have sporulated contains one or more substances which inhbit the germination of spores (akinetes) of this organism. Germination of spores occurs rapidly in the absence of these substances, but is virtually completely suppressed in their presence.     

Synthesis of poly(A)-containing RNA in outgrowing spores of Bacillus subtilis

The synthesis of poly(A)-containing RNA in outgrowing spores of Bacillus subtilis was studied. A significant amount of RNA puls-labelled with 3H-uridine is polyadenylated. With the beginning of RNA synthesis in outgrowing spores labelled poly(A)-containing RNA was detected. The amount of poly(A)-RNA during the outgrowth and first cell division remains constant. Besides poly(A)-RNA the synthesis of tRNA and rRNA occurs. These results indicate a simultaneous activation of synthesis of tRNA, rRNA as well as of poly(A)-containing RNA during outgrowth of B. subtilis spores.     

144 Use of Fourier Transform Infrared (FT IR) Diffuse Reflectance Spectroscopy and Hamming Distances to Study the Phycocolloid Chemosystematics of the Red Algae (Rhodophta)

Coalescence in seaweeds is known to occur in the laboratory among young and older sporelings and in the field between neighboring conspecific clumps. However, because spores and germlings are difficult to study in the field, it is as yet unknown at which stage of population development coalescence is most important. Since many seaweeds disperse aggregated propagules, often with a sticky mucilagous envelope around the spores, aggregated recruitment and coalescence might be more important at early stages of population establishment than among fully grown, well established clumps. Using recruitment plates maintained during several experimental times in the field, we are evaluating the above idea with mid-intertidal populations of Mazzaella laminarioides. During high fertility seasons, close to 45% of the spores settling within or at close (<1 m) distances of the bed exhibited aggregated recruitment, forming groups of 2 to 150+spores. The probability of aggregated recruitments is a function of dispersal distance and spore density. The number of sporelings produced is a function of spore density and coalescence. Highest after-recruitment mortality (first 15 days) occurs among solitary recruits, followed by sporelings conformed by small number of spores (2–4). Approximately 50% of the spores recruited, isolated or in group, coalesce within these 15 days, gradually forming massive sporelings with increasingly larger basal areas. Thus, after recruitment, sporelings may disappear (die), survive or coalesce. These three alternatives are integrated in a new demographic model for coalescing seaweeds (supported by grant FONDECYT 1020855).     

The heat resistance of bacterial spores and its relationship to the contraction of the forespore protoplasm during sporulation

The heat resistance of spores of 11 bacterial species is shown to correlate with the average decrease in volume of the protoplasm of spores that occurs during sporulation and that is measured from the stage in the development of the forespore at which the cortex can first be observed.     

Phycomycetes parasitizing the ovaries of blackflies (simuliidae)

The developmental stages of the fungus in the ovary and freshwater are discussed. Based on the evidence to date the simuliid host is infected in the early larval instars. Parasitism results in a destruction of the ovarian tissues with no or few eggs produced. The fungus replaces the ovarian tissue and produces massive numbers of spores which are released by the female fly at the time of oviposition. After oviposition, infected flies do not seek a blood meal. Longevity of the host does not appear to be affected by the presence of the fungus. The spores released by the fly during oviposition did not withstand desiccation and did not develop on the artificial media tested; development occurs only in water. Development of sporangia takes 5 to 7 days at 4°C, and 2 to 3 days at temperatures above 10°C. The developmental process in spores from S. verecundum was followed from the formation of sporangia to the opening of discharge tubes which zoospores were presumed to be released but not observed.     

QUALITATIVE AND QUANTITATIVE STUDIES OF THE SWIMMING BEHAVIOR OF HINCKSIA IRREGULARIS SPORES (PHAEOPHYCEAE)

The swimming behavior of spores of the brown alga Hincksia irregularis was analyzed using computer-assisted motion analysis. We distinguished five main swimming patterns: straight paths, search circles, orientation, gyration, and wobbling. We suggest different functional values for the individual swimming patterns. Straight paths, search circles, and orientation are different but all may be important in small-scale movements in the benthic boundary layer. As such, they could enable a spore to find a suitable microenvironment for germination and growth. Gyration occurs during the initial reversible phase of adhesion that can lead to settlement. Wobbling is typical for irritated or mechanically damaged spores and does not seem to be a typical pattern associated to settlement. The dominant swimming patterns changed with spore age (10 ± 5 to 60 ± 5 min of spore age), with young spores mainly swimming in straight paths and search circles and older spores in orientation and gyration. This change in swimming patterns can be quantified by speed (decrease over time) and rate of change of direction (increase over time). Based on these results, we suggest that computer-assisted motion analysis of the swimming behavior of H. irregularis spores can be used to develop bioassays with both ecological and technological relevance.     

An Examination of the Methods for Isolating Cellular Slime Molds (Dictyosteliida) from Soil Samples*

SYNOPSIS. Present methods for extracting cellular slime molds from soil samples are shown to be biased in favor of spores and microcysts. To alleviate this problem, a modified method is proposed together with a method for partitioning active (amebae) and inactive forms (spores and microcysts).     

Meiosis in a temperature-sensitive DNA-synthesis mutant and in an apomictic yeast strain (Saccharomyces cerevisiae).

It is shown that in the temperature-sensitive yeast mutant (Saccharomyces cerevisiae) spo 11 at the restrictive temperature of 34 degrees C. (1) premeiotic DNA synthesis is nearly completely blocked; (2) the nucleus enters meiotic prophase indicated by the formation of axial cores and polysynaptonemal complexes; (3) the kinetic apparatus functions normally at meiosis I and II; (4) early spore formation occurs in nearly all cells but it is variable and all spores eventually degenerate. It is concluded that chromosome replication is not a prerequisite for the functions listed above. The apomictic yeast strain 4117 produces 2 diploid spores. It is shown that a diploid which produces 2-spored asci, synthesized from 4117, no. 5, and an adenine requiring strain (1) has a normal meiotic prophase with abundant synaptonemal complexes; (2) has only one meiotic spindle; (3) has spores which form red clones more frequently than normal or u.v.-treated vegetative cells form ade/ade red sectors through mitotic recombination. It is concluded that this apomictic yeast has maintained meiotic prophase, but that one of the two meiotic divisions is suppressed.     

VALSA EUGENIAE IN RELATION TO THE SUDDEN-DEATH DISEASE OF THE CLOVE TREE (EUGENIA AROMATICA)

The sudden-death disease of the clove tree is invariably associated with the fungus Valsa eugeniae. The pathogenicity of this fungus has been investigated experimentally, and it has been shown that it is a primary parasite on mature clove trees, that saplings are resistant to it, and that seedlings are immune. It has been experimentally demonstrated that water-borne spores of the fungus can invade the absorbing and the fibrous roots of the clove tree. The slow-decline disease, which affects clove saplings only, is associated with a slow and progressive root-rot over a period of many years. Valsa is also invariably associated with this disease, which occurs only in areas which have been replanted after the previous stand has been killed by sudden death. It is thought that slow decline is the symptom-expression of Valsa attack on young trees when these still retain some measure of juvenile resistance to it.     

Paranucleospora theridion n. gen., n. sp. (Microsporidia,Enterocytozoonidae) with a Life Cycle in the Salmon Louse (Lepeophtheirus salmonis,Copepoda) and Atlantic Salmon (Salmo salar)

ABSTRACT. Paranucleospora theridion n. gen, n. sp., infecting both Atlantic salmon (Salmo salar) and its copepod parasite Lepeophtheirus salmonis is described. The microsporidian exhibits nuclei in diplokaryotic arrangement during all known life‐cycle stages in salmon, but only in the merogonal stages and early sporogonal stage in salmon lice. All developmental stages of P. theridion are in direct contact with the host cell cytoplasm or nucleoplasm. In salmon, two developmental cycles were observed, producing spores in the cytoplasm of phagocytes or epidermal cells (Cycle‐I) and in the nuclei of epidermal cells (Cycle‐II), respectively. Cycle‐I spores are small and thin walled with a short polar tube, and are believed to be autoinfective. The larger oval intranuclear Cycle‐II spores have a thick endospore and a longer polar tube, and are probably responsible for transmission from salmon to L. salmonis. Parasite development in the salmon louse occurs in several different cell types that may be extremely hypertrophied due to P. theridion proliferation. Diplokaryotic merogony precedes monokaryotic sporogony. The rounded spores produced are comparable to the intranuclear spores in the salmon in most aspects, and likely transmit the infection to salmon. Phylogenetic analysis of P. theridion partial rDNA sequences place the parasite in a position between Nucleospora salmonis and Enterocytozoon bieneusi. Based on characteristics of the morphology, unique development involving a vertebrate fish as well as a crustacean ectoparasite host, and the results of the phylogenetic analyses it is suggested that P. theridion should be given status as a new species in a new genus.     

Morphological and molecular characterization of a new microsporidian species from the predatory mite Metaseiulus occidentalis (Nesbitt) (Acari,Phytoseiidae)

A new microsporidian species is described from the predatory mite Metaseiulus (formerly Typhlodromus or Galendromus) occidentalis (Nesbitt) (Acari, Phytoseiidae). The ultrastructure of this new species is presented together with the first molecular characterization for a microsporidium of mites. All stages of this new microsporidium are haplokaryotic and develop in direct contact with the host-cell cytoplasm. Sporogony is disporoblastic and spores are formed in eggs, immature stages, and adults of M. occidentalis. There are two morphological classes of spores, one with a short polar filament (3-5 coils) that measured 2.53 x 1.68 microm and one with a longer polar filament (8-9 coils) that measured 3.14 x 1.77 microm. Horizontal transmission of this new species occurs by cannibalism of eggs and other stages and perhaps involves the spores with the long polar filament. Spores with the short polar filament may play a role in autoinfection and vertical (transovarial) transmission that is highly efficient in transferring the microsporidium from adults to progeny. Analysis of the small subunit ribosomal DNA indicated that this species from M. occidentalis is most closely related to the Nosema/Vairimorpha clade of microsporidia. A conflict between the morphological and molecular data is discussed. The species is compared to previously described microsporidia of arachnids resulting in creation of Oligosporidium occidentalis n. sp. in the family Unikaryonidae.     

Ultrastructure of the microsporidian Inodosporus octospora (Thelohaniidae), a parasite of the shrimp Palaemon serratus (Crustacea, Decapoda)

A parasite of the muscle of the shrimp Palaemon serratus has been examined by light and electron microscopy. Development occurs among myofibrils and induces ultrastructural alterations of the muscle fibers causing white discoloration. This microsporidian is characterized by uninucleate, later diplokaryotic and di-diplokaryotic meronts. The mother cell develops by rosette-like budding into 8 uninucleate sporoblasts, each containing 3 tape-like filaments attached to the wall that is enclosed in a persistent sporophorous vacuole. Each sporoblast gives rise to a uninucleate spore that possesses 3 elongated tape-like filaments attached to the spore wall, like spore tails. The morphological characters of the spores, redescribed in the present study, suggested that the spores belonged to Inodosporous octospora. The possibility that in the future members of Inodosporus sp. may be considered a new parasite group is discussed.     

EXTERNAL SPORE MORPHOLOGY AND TAXONOMIC AFFINITIES OF PHYLLOGLOSSUM DRUMMONDII KUNZE (LYCOPODIACEAE)

The external spore morphology of Phylloglossum drummondii was studied with the scanning electron microscope and shown to share a number of features with spores in the subgenus Urostachys in Lycopodium. Within Urostachys the affinities of P. drummondii were less evident: the foramino-fossulate distal face, pyramidal proximal surface, and subcircular outline of its spores are similar to spores in the section Phlegmaria while the weakly foveolate proximal surface and raised laesurae are characteristic of spores in the section Selago.     

“INTERNAL THECAE” OF EUNOTIA SOLEIROLII (BACILLARIOPHYCEAE): DEVELOPMENT,STRUCTURE AND FUNCTION AS RESTING SPORES1

The “double thecae” or “internal septa” of Eunotia soleirolii (Kütz.) Rabenh, are shown to represent the thecae of resting spores, as characterized by their physiology, as well as morphology. They differ from all resting spores of centric diatoms by the formation of both their valves as a result of unequal cell divisions; and, from the majority of centric spores by the presence of several girdle bands in both their thecae. Spore formation can be induced by high or low pH, high temperature (24 C), and iron, silica, phosphate or nitrate deficiencies, whereas low temperatures defer it. Spores do not germinate directly, but dormancy can be removed by dark treatments (–2 to 15 C) for a minimum of 4–5 wk. Longer dark treatments result in higher germination rates. At 15 C, a minimum of 2 mo is required and 4 mo is better. Heat treatments (27–42 C) are ineffective, but may shorten the dormancy-breaking subsequent cold period. Instances of secondary dormancy, as well as relative dormancy, were observed. Germination usually occurs in the light between 2 and 21 C. An equal division of the spore is followed by unequal divisions of both new cells with only the two resulting large cells being viable. The experiences in the laboratory aided the discovery of stages of spore germination in nature.     

Die Steuerung der Brandsporenkeimung und Sporidienbildung bei Tilletia caries (DC) Tul. durch Licht

Zusammenfassung Durch Licht wird bei Tilletia caries (DC) Tul. nicht nur die Keimungsgeschwindigkeit, sondern auch der Endwert der Keimung erhöht.Es konnte gezeigt werden, daß sowohl die Promycel- als auch die Sporidienbildung durch Licht gesteuert wird.Während es sich bei der Beeinflussung der Promycelbildung um eine induktive Lichtwirkung handelt, muß für eine maximale Förderung der Sporidienbildung während des entsprechenden Entwicklungsabschnittes Dauerlicht hoher Intensität geboten werden.Licht ist erst 3 Tage nach Sporenaussaat wirksam. Die Beeinflussung der Promycelbildung ist am stärksten, wenn das Licht zwischen dem 3. und 6. Tag geboten wird, während der Prozentsatz der Sporidienbildung um so größer ist, je später der Zeitpunkt der Belichtung liegt.Wie bei nahezu allen Photomorphosen bei Pilzen besitzt Blaulicht die stärkste Wirksamkeit. Grün ist zwar ebenfalls noch eindeutig wirksam, während zwischen dem Rotwert und der Dunkelkontrolle kein signifikanter Unterschied besteht.

---

The role of light in spore germination and sporidial production of the wheat smut fungi Tilletia caries (DC.) Tul.

Summary In Tilletia caries, light hastens the beginning of spore germination and increases the maximum percentage of spores which germinate.It could be shown, that light is required for two different steps—the formation of germ tubes and the sporidial development.The stimulation of the spore germination is a photoinductive process; but the best sporidial development occurs in continuous light of high intensity.There is a sensitive period in the response of spores to light. High germination is induced when the light exposure occurs between the 3rd and 6th day after the spores were sowed on the medium. Best sporidia production results when the spores were irradiated at the 5th day or later.As in nearly all photoresponses in fungi, blue light is more effective than green. No significant difference exists between the red light and dark experiments.