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.     

Infrapopulations of Sclerocollum saudii Al-Jahdali, 2010 (Acanthocephala: Cavisomidae) in the rabbitfish Siganus rivulatus (Teleostei, Siganidae) from the Saudi coast of the Red Sea

In infrapopulations of helminth parasites, density-dependent effects, through some form of intra- and interspecific competition, play an important role in shaping and regulating the infrapopulations. The mechanisms responsible for these processes have often been observed in laboratory studies and rarely studied under natural conditions. Here, 24 natural infrapopulations (77-447 individuals) of the acanthocephalan Sclerocollum saudii Al-Jahdali, 2010 from the fish Siganus rivulatus consisted of cystacanths, newly excysted juveniles, immature and mature worms, distributed in a well-defined fundamental niche (anterior 60% of the intestine). Each of these stages exhibited a significantly different longitudinal distribution within this niche. In small infrapopulations, cystacanths and newly excysted juveniles were found in the sixth 10% of the intestine, immature worms in the fifth 10% and mature worms in the anterior 40% of the intestine. However, their proportions followed a clear ascending order in each infrapopulation, and the female-male ratios of both immature and mature worms were distinctly female-biased. In large infrapopulations, mature worms existed partially in the site of immature ones, where a differential mortality among immature females was constantly observed. However, the proportions of immature worms increased significantly and those of mature worms decreased significantly, the mean lengths of immature and mature females decreased dramatically and the female-male ratios were distinctly male-biased. The mean sizes of immature and mature males seemed stable through all infrapopulations. The distribution of mature males and females suggests intense male-male competition for access to females, and reveals that larger females are copulated prior to the smaller ones. The results are statistically significant and suggest that infrapopulation self-regulation is through density-dependent mechanisms, in which immature females may play a key role.     

A new species of Haplosporidium Caullery & Mesnil, 1899 in the marine false limpet Siphonaria lessonii (Gastropoda: Siphonariidae) from Patagonia

A new species of Haplosporidium Caullery & Mesnil, 1899 parasitising the pulmonate gastropod Siphonaria lessonii Blainville in Patagonia, Argentina, is described based on morphological (scanning and transmission electron microscopy) and sequence (small subunit ribosomal RNA gene) data. Different stages of sporulation were observed as infections disseminated in the digestive gland. Haplosporidium patagon n. sp. is characterised by oval or slightly subquadrate spores with an operculum that is ornamented with numerous short digitiform projections of regular height, perpendicular to and covering its outer surface. The operculum diameter is slightly larger than the apical diameter of the spore. Neither the immature nor mature spores showed any kind of projections of the exosporoplasm or of the spore wall. Regarding phylogenetic affinities, the new species was recovered as sister to an undescribed species of Haplosporidium Caullery & Mesnil, 1899 from the polychaete family Syllidae Grube from Japanese waters. The morphological characters (ornamentation of the operculum, spore wall structure, shape and size of spores, and the lack of spore wall projections) corroborate it as an as yet undescribed species of Haplosporidium and the first for the phylum in marine gastropods of South America. Siphonaria lessonii is the only known host to date.     

Henneguya adherens N. Sp. (Myxozoa, Myxosporea), Parasite of the Amazonian Fish, Acestrorhynchus falcatus

ABSTRACT. A new species of myxosporean from the gill filaments of the freshwater teleost fish, Acestrorhynchus falcatus collected in the Amazon river is described from light and transmission electron microscope observations. The mature spores (total length 32.3 [30.7–35.1] μ) and all developmental stages were found in the same sporogonic plasmodium. The ellipsoidal spore body consists of 2 unequal shell valves adhering together along the suture lines. Each valve, tapering as a caudal projection, forms a long tail (length 20.5 [18.0–21.7] μm). The tail was surrounded by a homogeneous sheath on its length. The polar capsules measuring 3.1 × 1.2 μm contain 3–4 coils of the polar filament. All surfaces of the immature and mature spores were surrounded by a closely adherent homogenous structural sheath, mainly thicker around the tails. The taxonomic affinities of this parasite to other species are discussed.     

Examination of the cyst of Malamoeba locustae and of the spore of Mattesia dispora with the scanning microscope

The fine structure of the surface of cysts of the amoeba Malamoeba locustae and that of spores of the neogregarine Mattesia dispora have been examined with the scanning microscope. Freeze-drying was found to be the most convenient of the three methods tested for the preparation of the specimens. The surface of both mature and immature cysts of M. locustae is smooth, without a microrelief. By contrast, spores of M. dispora are equipped with a small, circular structure at the poles indicating the site of the future pore through which the sporozoites will emerge in the intestine of the new host. In addition, a flat relief forming small depressions is discernable on the surface of these spores at a high magnification.     

团毛菌目粘菌个体发育比较研究

利用有饲培养方法,对团毛菌目粘菌的盖碗菌属、团网菌属、半网菌属、变毛菌属及团毛菌属 7个种的个体发育进行了比较研究。描述了其原质团发育特征及小胞囊的特点,并将团毛菌目粘菌原质团划分为两种类型,即原始中间型和发达中间型。同时利用光学显微镜及电子显微镜观察描述了子实体的主要发育特征。结果表明,孢丝或孢子表面纹饰不明显的种类较原始,表面纹饰明显的种类为发达类型。     

Nadelspora canceri N. G., N. Sp., an Unusual Microsporidian Parasite of the Dungeness Crab, Cancer Magister

ABSTRACT. The microsporidium Nadelspora canceri n. g., n. sp., is described from the striated musculature of the Dungeness crab ( Cancer magister ) in Oregon, USA. The needle-shaped spores were rounded anteriorly, tapered to a posterior point and measured 7.1–11.8 × 0.2–0.3 μm in fixed preparations. The extremely narrow spore diameter prevented observation of morphological details at the light microscopic level and ultrastructural details of mature spores were difficult to resolve. Meronts were not observed and the monokaryotic merozoites and sporonts were not contained within either parasitophorous or sporophorous vesicles. Sporonts were disporoblastic and gave rise to monokaryotic sporoblasts that became narrow and elongate as they developed into immature spores with a developing polar filament. The nucleus was not clearly resolved in mature spores and may have been surrounded by the lamellar polaroplast. The polar filament was of nearly uniform diameter throughout most of its length and ended abruptly about three-fourths of the distance from the anterior end of the spore. Unusual spherical non-membrane bound granules surrounded the polar filament in a spiral arrangement. The new microsporidium resembles members of the family Mrazekiidiae, but differs in lacking a diplokaryon at any stage. It is probably most closely related to Baculea daphniae from which it differs primarily by spore shape and size. The familial relationships of the genus Baculea have not been determined and it is proposed to include it with Nadelspora in the new family Nadelsporidae.     

Ultrastructural study of the late stages of Loma salmonae development in the gills of experimentally infected rainbow trout

The main objective of this investigation was to examine the ultrastructural features of gills from rainbow trout experimentally infected with Loma salmonae to determine the morphological events that occur during the late stages of development of this parasite. Peripheral distribution of the mature parasites inside round xenomas was observed at weeks 5 and 6 postexposure (PE), but eventually the parasite occupied the entire xenoma. Degenerative changes were observed only in immature parasites at week 7 PE, and eventually an inflammatory reaction with a cellular infiltration was directed against mature spores. Round, flattened, and irregular shaped xenomas were observed at week 8 PE. The round xenomas showed a severe inflammatory response with disintegration of the xenoma membrane. This event was accompanied by eversion of polar tubes within the attacked xenoma and by the simultaneous presence of 2 tubular appendages, the type I and II tubules. Flattened xenomas were observed below the endothelium of gill lamella arteries. The irregular xenomas were located in the connective tissue of the gill filament and showed multiple projections occupied by spores. Both flattened and irregular xenomas showed no evidence of inflammatory reaction. An earlier proposed hypothesis is expanded to explain how L. salmonae is implanted beneath lamellar endothelium and within filament connective tissue.     

Ultrastructure of Enteromyxum leei (Diamant, Lom, & Dyková, 1994) (Myxozoa), an Enteric Parasite Infecting Gilthead Sea Bream (Sparus aurata) and Sharpsnout Sea Bream (Diplodus puntazzo)

ABSTRACT. The ultrastructure of the developmental stages of the myxozoan Enteromyxum leei parasitizing gilthead seabream ( Sparus aurata ) intestine and sharpsnout sea bream ( Diplodus puntazzo ) intestine and gallbladder are described. The earliest stage observed was a small dense trophozoite located among enterocytes. Proliferative stages, observed intercellularly in the epithelium of the intestine and gallbladder as well as in the lumen, possessed the typical cell-in-cell configuration throughout their development. Secondary cells were seen undergoing division within a common vacuolar membrane that also encompassed pairs of tertiary cells. Cytochemical studies showed that primary cells stored mainly lipids whereas secondary cells stored abundant β-glycogen granules. Sporogonic development resembled that described for other disporous myxozoans. Within sporogonic stages, nonsporogonic secondary cells were observed accompanying two developing spores. Mature spores had a binucleated sporoplasm in which glycogen stores were abundant and no sporoplasmosomes were found. Our observations are discussed in relation to our knowledge on other myxozoans of the genus Enteromyxum .     

Debris cloaking in Endomychidae: a new species from Peru (Goleoptera)

Adults and immature stages of the mychothenine endomychid Bystus decorator sp. nov. from Peru are described. All life stages occur together on rotting logs and feed on fungal spores. Larval debris cloaking and morphological adaptations associated with it are discussed in detail. Some larval setae are apically clubbed and barbed along the shafts to facilitate particle adhesion. Pronation of the thoracic lateral lobes bring the setae into contact with the substrate to gather particles of debris. The anatomy and functional morphology of the thoracic musculature is considered in the context of cloaking behaviour. Variations in debris cloaking behaviours among mychothenine endomychids are discussed and compared to other debris-cloaking insects.     

Ultrastructure of Enteromyxum leei (Diamant, Lom, & Dyková, 1994) (Myxozoa), an enteric parasite infecting gilthead sea bream (Sparus aurata) and sharpsnout sea bream (Diplodus puntazzo).

The ultrastructure of the developmental stages of the myxozoan Enteromyxum leei parasitizing gilthead seabream (Sparus aurata) intestine and sharpsnout sea bream (Diplodus puntazzo) intestine and gallbladder are described. The earliest stage observed was a small dense trophozoite located among enterocytes. Proliferative stages, observed intercellularly in the epithelium of the intestine and gallbladder as well as in the lumen, possessed the typical cell-in-cell configuration throughout their development. Secondary cells were seen undergoing division within a common vacuolar membrane that also encompassed pairs of tertiary cells. Cytochemical studies showed that primary cells stored mainly lipids whereas secondary cells stored abundant beta-glycogen granules. Sporogonic development resembled that described for other disporous myxozoans. Within sporogonic stages, nonsporogonic secondary cells were observed accompanying two developing spores. Mature spores had a binucleated sporoplasm in which glycogen stores were abundant and no sporoplasmosomes were found. Our observations are discussed in relation to our knowledge on other myxozoans of the genus Enteromyxum.     

Ultrastructure of Myxobolus brycon n. sp. (Phylum Myxozoa), parasite of the piraputanga fish Brycon hilarii (Teleostei) from Pantanal (Brazil)

Light and electron microscopy studies of a myxosporean, parasitizing the gill filaments of the freshwater fish Brycon hilarii (Valenciennes, 1850) (Characidae) collected in the Paraguay River (18°49'S, 57°39'W) (Pantanal), Brazil, is described. This parasite produces spherical to ellipsoidal polysporic histozoic plasmodia (Pmd) (up to ～180 μm in diameter) delimited by a double membrane and with several pinocytic channels. The plasmodial cyst contained the youngest developmental stages at the cortical periphery and immature and mature spores more internally. The Pmd developed near the cartilaginous structure of the gill filament, forming a prominent deformation where the gill lamellae disappear. Pyriform spores measured 6.9±0.6 (range 6.5-7.2) μm long, 4.2±0.5 (range 3.9-4.8) μm wide, and 2.5±0.7 (range 1.9-2.8) μm thick. The spores composed of two equal shell valves (～70 nm thick), adhering together along the straight suture line, surrounded two equal symmetric and elongated to pyriform polar capsules (PC) 4.2±0.6 (range 3.8-4.7) × 1.9±0.6 (1.7-2.5) μm; each PC contained a coiled polar filament with eight or nine (rarely 10) turns and a binucleated sporoplasm cell. Dense irregular masses were observed among the polar filaments coils. An intercapsular appendix was not observed. The sporoplasm contained several globular sporoplasmosomes randomly distributed among an extensive rough endoplasmic reticulum system with numerous vesicles and cisternae. Based on the morphological and ultrastructural differences and specificity of the host, we establish the new species, Myxobolus brycon n. sp.     

New species of Parathelohania (Microsporidia) in larva of the Anopheles aquasalis (Diptera: Culicidae) in Venezuela

The symptoms of Anopheles aquasalis larvae naturally infected by a microsporidium, and the ultrastructure of the infecting spores is described. The larvae were maintained under laboratory conditions in salt concentrations of 10 g/l and 20 g/l of water. Daily recordings of the mortality of the larvae were made and the dates of change of instar were noted. Infected larvae were processed for transmission electronic microscopy using conventional methods at pH 7.2 and 260 mOsm/l. The infection by the microsporidia was positively correlated with an increase in the mean duration of the fourth instar of 2.88 to 6.33 days in 10 g/l of salt and of 2.47 to 6.14 days in 20 g/l of salt. Larval mortality also increased by approximately 50% during this instar in both salt concentrations. Development time and survival were not affected during the other immature stages. The mature spores found in the intestines of infected larvae were barrel shaped and measured approximately 2.6 x 2.4 mm. The exospore has a collar shaped prolongation at the posterior end of the spore. The spores are uninuclear with a posterior vacuole. The polar filament is anisofilar with nine rings, five with a diameter of 58 nm each and four with a diameter of 23 nm each. The polarplast is lamellate, and more tightly packed in the apical region. The reduction of the survival of A. aquasalis larvae infected with the microsporidia, and the increase in the development time suggest that this parasite might have a potential as a biological control of this pest. The microsporidium described here has similar characteristics to that of the genus Parathelohania. I suggest that the microsporidium found in A. aquasalis represents a new species and I propose the name Parathelohania aquasalensis. This is the first report of a microsporidium from a dipteran in Venezuela.     

Description of Hyalinocysta expilatoria n. sp., a microsporidian parasite of the blackfly Odagmia ornata

Hyalinocysta expilatoria n. sp. is described from a larva of Odagmia ornata collected in Sweden. Infection was restricted to the adipose tissue which was transformed into a syncytium. The earliest stage observed was diplokaryotic merozoites, which mature directly into diplokaryotic sporonts. Each sporont produces a sporophorous vesicle (pansporoblast), which persists, also enclosing mature spores. Usually nuclear divisions result in a plasmodium with 8 nuclei, which fragments into 8 sporoblasts, each of which develops into a spore without further division. Occasionally an aberrant number of spores (2, 4, 6) is formed. The spores are pyriform with a flattened area at the posterior pole. Spores in sporophorous vesicles with 8 spores are 4.0–6.0 μm long, in vesicles with 4 spores 4.0–5.0 μm, and in vesicles with 2 spores 7.0–8.0 μm. In some vesicles the spores develop asynchronously, and 2, 4, or 6 mature spores are found together with 6, 4, or 2 immature. There was also a small number of vesicles with supernumerary spores, less than 8 normally developed. The 325–350 nm thick spore wall is composed of three layers. The polar filament is anisofilar with 7 coils in a single layer. The anterior 5–6 coils are wide, the posterior 2-1 thin. The angle of tilt of the anterior filament coil is approximately 50°. The spore has a single nucleus. The sporophorous vesicle is delimited by a thin membrane, also visible in haematoxylin stained preparations. Vesicles with mature spores are void of metabolic inclusions.     

Ultrastructural data on the spore of Myxobolus maculatus n. sp. (phylum Myxozoa), parasite from the Amazonian fish Metynnis maculatus (Teleostei)

Light and electron microscopy studies of a myxosporean, parasitic in the intertubular interstitial tissue of the kidney of the freshwater teleost fish Metynnis maculatus Kner, 1860 (Characidae) from the lower Amazon River (Brazil), are described. We observed polysporic histozoic plasmodia delimited by a double membrane and with several pinocytic channels and containing several life cycle stages, including mature spores. The spore body was of pyriform shape and was 21.0 microm long, 8.9 microm wide and 7.5 microm thick. Elongated-pyriform polar capsules were of equal size (12.7 x 3.2 microm) and contained a polar filament with 14 or 15 coils. The spore features fit those of the genus Myxobolus. Densification of the capsular primordium matrix, which increased in density from the inner core outwards, differentiating at the periphery into small microfilaments measuring 45 nm each, and tubuli arranged in aggregates and dispersed within the capsular matrix of the mature spores, are described. Based on the morphological differences and specificity of the host, we propose the creation of a new species named Myxobolus maculatus n. sp.     

NUCLEAR MIGRATION AND ASYMMETRIC CELL DIVISION IN ONOCLEA SENSIBILIS SPORES: AN ULTRASTRUCTURAL AND CYTOCHEMICAL STUDY

A method of preparation for electron microscopy of fern spores in early stages of germination is presented. The cytochemistry and fine structure of Onoclea spores during the early stages of germination are described. The cytoplasm of the hydrated spore is filled with lipid droplets, protein granules and chloroplasts. During the early stages of development ribosomes and mitochondria increase in the area surrounding the central nucleus, and a new peripheral wall forms around the protoplast. Microtubules and large, branching mitochondria are associated with the nucleus during migration from its original central position in the spore to the proximal face and then to one end of the spore. There is no morphological polarization of cytoplasmic organelles of the spore before migration of the nucleus.     

Ultrastructure of neurosecretory cells in the frontal ganglion of the tobacco hornworm, Manduca sexta(L)

Ultrastructural evidence is presented to indicate the synthesis, storage, and packaging of neurosecretory material in the frontal ganglion of Mandaca sera. Two morphologically distinct stages in the development of NSG were observed in the NSC. The immature granule contains granular electron dense material and arises from dilations of the rough endoplasmic reticulum. In diapause pupae the immature granules accumulate, whereas in developing larvae and pupae, they fuse with the proximal face of the Golgi apparatus. The mature granule contains opaque electron dense material and is seen in close association with the distal cisternae of the Golgi apparatus in developing pupae and larvae. The mature granules presumably contain the material from the immature granules which has been chemically altered and condensed by the Golgi apparatus and probably represent the mature NSG.The second process in neurosecretory cells of developing larvae is cyclic and the various stages are described. A comparison of neurosecretion in photoperiodically induced diapause and developing (non-diapause) larvae and pupae is presented.     

A new microsporidian parasite, Potaspora morhaphis n. gen., n. sp. (Microsporidia) infecting the Teleostean fish, Potamorhaphis guianensis from the River Amazon. Morphological, ultrastructural and molecular characterization

A fish-infecting Microsporidia Potaspora morhaphis n. gen., n. sp. found adherent to the wall of the coelomic cavity of the freshwater fish, Potamorhaphis guianensis, from lower Amazon River is described, based on light microscope and ultrastructural characteristics. This microsporidian forms whitish xenomas distinguished by the numerous filiform and anastomosed microvilli. The xenoma was completely filled by several developmental stages. In all of these stages, the nuclei are monokaryotic and develop in direct contact with host cell cytoplasm. The merogonial plasmodium divides by binary fission and the disporoblastic pyriform spores of sporont origin measure 2.8+/-0.3 x 1.5+/-0.2 microm. In mature spores the polar filament was arranged into 9-10 coils in 2 layers. The polaroplast had 2 distinct regions around the manubrium and an electron-dense globule was observed. The small subunit, intergenic space and partial large subunit rRNA gene were sequenced and maximum parsimony analysis placed the microsporidian described here in the clade that includes the genera Kabatana, Microgemma, Spraguea and Tetramicra. The ultrastructural morphology of the xenoma, and the developmental stages including the spores of this microsporidian parasite, as well as the phylogenetic analysis, suggest the erection of a new genus and species.     

Biochemical genetics of bacterial sporulation

Summary The morphological and biochemical characters of twenty nine sporulation mutants were compared. Some of the predictions made on the basis of unidirectional pleïotropic interactions were confirmed, namely that the latest proteolytic enzymes, like elastase, are related to late morphological stages. From the cytological point of view, mutants blocked at various stages were described. Among the late mutants, both coatless mutants with normal but incomplete cortex and cortexless mutants with flexible spore coats were found. Particularly interesting is the class of abnormal late sporulation mutants which form normal mature heat-resistant spores at high frequencies, but, in addition, present various anomalies in the structure of the spore coats and various sporangial inclusions such as a spongy fibrous material, resembling the cortex, and either onion-like or rod-shaped inclusions, probably formed by spore coat components. The presence of these structures is related to the derepression of elastase activity and may reflect overproduction of spore components. Several mutants also contain abnormal, large, dark, membrane-bound mesosomes, either compact or loose, whose presence is related to the lack of oxidation of tetrazolium dyes. The morphological heterogeneity of mutant populations is also noted. These findings are discussed in relation to the theory of sequential gene activation.