[THE MICROSPORIDIUM GLUGEA GASTEROSTEI VORONIN 1974 (MICROSPORIDIA: MARINOSPORIDIA) FROM THE THREE-SPINED STICKLEBACK GASTEROSTEUS ACULEATUS (ACTINOPTERYGII: GASTEROSTEIFORMES) AS AN INDEPENDENT SPECIES].

The microsporidium Glugea gasterostei from the three-spined stickleback Gasterosteus aculeatus was described as an independent species basing upon morphological and ecological traits of the parasite (Voronin, 1974), further supported by ultrastructural characters of its spores (Voronin, 1983). During the revision of microsporidia of the genus Glugea (Canning, Lom, 1986; Lom, 2002), the validity of this species was doubted and it was synonymized with G. anomala. Nevertheless, the molecular phylogenetic analysis performed in the present study showed the unique molecular haplotype of small subunit rRNA gene of G. gasterostei (Genbank accession number KM977990) and its close relatedness to G. anomala, G. atherinae and G. hertwigi (sequence similarity of 99.7 %). One of typical characters of G. gasterostei, as opposed to G. anomala, is the formation of xenomas on inner tissues and not on the surface of infected fishes. This feature is retained even after the infection of different host species. Taken together, these data confirm the validity of G. gasterostei as a separate species among closely related taxa that had diverged comparatively recently.

[Should the microsporidian spores be treated as dormant stages?].

Spores of bacteria, fungi, microsporidia and other protists are traditionally treated as dormant stages, intended to the long-term survival in the environment and to activation of parasitic forms during the infestation of a new host. However, in the process of examination of insect microsporidia at the molecular cellular levels and also at the level of organisms and populations, we came to a conclusion that spores are very active developmental stages with the entire potential directed to the rapid and successful infestation of new hosts during contact with the later. The work summarizes the original data demonstrating (1) the necessity of the rapid activation of microsporidian spores during host contact, (2) hopelessness of the long retaining of viability by spores of many microsporidia in the environment after leaving host organism; and (3) specific accumulation of metabolic ferments in "dormant" spores, but not in actively proliferating prespore developmental stages. On the basis of these data we conclude that microsporidian spores tend to shorten the period when they stay outside host organism to the maximal degree. The probability of host infestation within the limited time period increases due to diverse modes of transmission of pathogens, accumulation of maximally possible volume of infective spores, and the rapid mobilization of the extrusion apparatus.

[Crepidulospora--nomen novum for the junior generic homonym (preoccupied generic name) Crepidula]. / Crepidulospora--novoe nazvanie dlia mladshego gomonima, (preokkupirovannogo rodovogo imeni) Crepidula.

Crepidulospora nom. nov. is a replacement generic name for the genus Crepidula Simakova, Pankova et Issi, 2003 based on the type species Crepidula beklemishevi Simakova, Pankova et Issi, 2003 (Microsporida) from Anopheles beklemishevi. The name proposed by Simakova et al., 2003 is a preoccupied name, because it was already used for the gastropode Crepidula Lamarque 1899 (Echinospirida, Calibraeidae), a parasite of Mytilus. A valid name of the type species of the genus is now Crepidulospora beklemishevi (Simakova, Pankova, Issi, 2003) comb. n. Crepidulospora Simakova, Pankova et Issi nom. non. Type species: Crepidula beklemishevi Simakova, Pankova, Issi, 2003. Diagnosis. Sporogony is octosporoblastic. Sporogonal stages are in direct contact with host cell cytoplasm. 8 uninucleate spores, 4.2 x 2.2 mkm, are sandals-like. Polar tube is anisofilar, with 6-7 coils (2 + 4-5). Polaroplast is three-partite, with broad vesicular, vesicular and lamellar compartments. Microsporidia ilnfects larval adipose tissues. Type host: Anopheles beklemishevi (Diptera, Culicidae).

[Crepidula beklemishevi gen. et sp. n. and Dimeiospora palustris gen. et sp. n. (Microspora: Amblyosporidae)--new microsporidian genera and species from blood-sucking mosquitoes (Diptera: Culicidae) from the south of the western Siberia]. / Crepidula beklemishevi gen. et sp. n. i Dimeiospora palustris gen et sp. n. (Microspora: Amblyosporidae)--novye mikrosporidii iz krovososushchikh komarov (Diptera: Culicidae) iuga zapadnoi Sibiri.

Microsporidia parasitizing the adipose body of mosquito larvae of Anopheles beklemishevi and Aedes punctor has been studied. Two new genera of microsporidia are described based on lightmicroscopic and ultrastructural characteristics of spores and sporogony stages. The spore wall of Crepidula beklemishevi gen. n. et sp. n. is formed by two-membrane exospore, thick exospore, bilayer endospore and thin plasmolemma. Spores with single nucleus, polar filament anisofilar, with 6-7 coils (2+ 4-5), polaroplast consisting of three parts: macrochelicoidal, microhelicoidal and lamellar. Fixed spores 4.2 +/- 0.22 x 2 +/- 0.01 microns. The sporogony of Dimeiospora palustris gen. et. n. results in spore formation of two different types. Spores of the first type are oviform, with thick wall, single-nuclear, 6.1 x 4.9 microns. Spore wall with three layers, about 370 nm. Exospore electron-dense, subexospore moderately electrondense. Exospore and subexospore irregularly pleated on the almost spore surface and slightly thinner on anterior end only. Endospore electron-translucent. Polar filament anisofilar, with 9 coils (3 + 6). Polaroplas consists of three parts: lamellar, fine bubbled, and coarse bubbled. Spores of the second type broad-ovate, with apical pole narrower, distal pole concave, 4.6 x 3.7 microns. Spore wall with three layer, 355 nm. Exospore on the apical end irregularly pleated, consists of thin electrondense exospore, subexospore of variable electron density, endospore electron-translucent. Polar filament anisofilar, with 13 coils (3 + 10). Polaroplast has two parts: lamellar and vesicular.

Establishment of the new genus Paranosema based on the ultrastructure and molecular phylogeny of the type species Paranosema grylli Gen. Nov., Comb. Nov. (Sokolova, Selezniov, Dolgikh, Issi 1994), from the cricket Gryllus bimaculatus Deg.

The ultrastructure of the microsporidian parasite Nosema grylli, which parasitizes primarily fat body cells and haemocytes of the cricket Gryllus bimaculatus (Orthoptera, Gryllidae) is described. All observed stages (meront, meront/sporont transitional stage ("second meront"), sporont, sporoblast, and spore) are found in direct contact with the host cell cytoplasm. Nuclei are diplokaryotic during almost all stages of the life cycle, but a brief stage with one nucleus containing an abundance of electron-dense material is observed during a "second merogony." Sporogony is disporous. Mature spores are ovocylindrical in shape and measure 4.5+/-0.16micromx2.2+/-0.07 microm (n=10) on fresh smears and 3.3+/-0.06 micromx1.4+/-0.07 microm (n=10) on ultrathin sections. Spores contain 15-18 coils of an isofilar polar filament arranged in one or two layers. Comparative phylogenetic analysis using rDNA shows N. grylli to be closely related to another orthopteran microsporidian, Nosema locustae, and to Nosema whitei from the confused flour beetle, Tribolium confusum. Antonospora scoticae, a parasite of the communal bee Andrena scotica, is a sister taxon to these three Nosema species. The sequence divergence and morphological traits clearly separate this group of "Nosema" parasites from the "true" Nosema clade containing Nosema bombycis. We therefore propose to change the generic name of N. grylli and its close relative N. locustae to Paranosema n. comb. We leave N. whitei in former status until more data on fine morphology of the species are obtained.

[Impact of microsporidia on hormonal balance in insect hosts]. / Vliianie mikrosporidii na gormonal'nyi balans nasekomykh.

Microsporidia (M) is a phylum of protists parasitizing obligatory in animal cells. Long way of adaptation of M to intracellular parasitism resulted in establishment of quite close relationships between the parasite and its host. Different species of M induce in their hosts symptoms similar to those caused by misbalance of juvenile hormone (JH) and ecdysone. M infection leads to pathology of different hormone-dependent functions such as cell differentiation and specialization, molting, metamorphosis, diapause and reproduction of insects. The signs of hormonal dysfunction evidence for elevated titer of JH in M-infected insects. Two possible explanation of this could be offered: JH secretion by M or specific influence of the parasites on the insect endocrine systems. Impact on insect endogenous JH titer by M could be mediated by affection of secretory activity of corpora allata or by suppression of enzymatic degradation of JH. According to different hypotheses, insect hormonal status during microsporidiosis could be modified by a) insect host stress-reaction, b) exhaustion of insect host reserves, characteristic for acute phase of the disease, c) destruction of infected insect cells and tissues during mass sporogenesis of M. Data found in literature and provided by our experiments evidence for presence of JH analogues or juvenilizing substance in the extracts of M spores. From detailed examination of pathological process it is also seen that juvenilizing effect of M infection is usually restricted to the invaded regions of tissues (i.e. expressed locally) but not a systemic one. Ability of M to modify morpho-functional features of infected tissues at the level of hormonal regulation is undoubtfully a prominent adaptation for stabilizing "microsporidia-insect" parasite-host systems.

[Parasitic systems of Microsporidia: descriptions and terminology questions]. / Parazitarnye sistemy mikrosporidii: opisanie i voprosy terminologii.

Three parasitic systems of Microsporidia are described: the system of monoxenic Vairimorpha mesnili with paraxenic hosts presented lepidopteran and hymenopteran species; the system of dixenic Amblyospora sp. with metaxenic hosts presented bloodsucking mosquitoes and crustaceans and the system of Metchnikovella sp. as parasite of other obligate parasite. The last case is characterized by very intimate interrelations between hyperparasite (microsporidian species), obligate parasite--host of Microsporidia (gregarine) and hyperhost--host of gregarine (polychaeta). This hyperparasite system is exclusive case of parasitic systems. Parasitic and hyperparasitic systems reflects a population level of host-parasite interactions. On biocenotic level many other organisms such as predators, vectors and reservators of invasion stages of Microsporidia affect parasitic systems giving a chance to one of the members of the system (to the host or to the parasite). These organisms form epiparasitic system. In all cases of the parasitic systems there are two-way communications between parasites and their hosts. In systems on biocenotic level--parasitic consortium--members of epiparasitic systems acts on parasitic systems, but members of parasitic systems don't affect epiparasitic systems.

[Refinement of the diagnosis of Issia globifera (Microsporidia: nosematidae) based on ultrastructural data]. / Utochnenie diagnoza Issia globulifera (Microsporidea, nosematidae) na osnove dannykh po ul'trastrukture mikrosporidii.

An ultrastructure of a microsporidian belonging to the aberrant and poorly studied genus Issia is discribed in this paper. The first description of Issia globulifera (Issi, Pankova, 1983) was made on the basis of light microscopic analysis of midgut smears of the infected insects. Both late merogonic and sporogonic stages can be met on the sections from midguts of Anopheles messae. Meronts are ameba-like cells (3.0-3.5 x 2.0-2.5 microns) forming conglomerates of cells without any regular organization. Meronts and sporonts directly contact with the host cell cytoplasm; they are surrounded by numerous mitochondria. Sporonts divide into 2 sporoblasts, but remain connected with each other by posterial ends. In the vicinity of the contact the paramural bodies (structures, presumably participating in the formation of the exospore) can be seen. Sporoblast morphogenesis is accompanied by the rapid grow of electron dense layer in the region of the contact of two sister sporoblasts. Due to such intensive growth the sporoblasts are finally located at sharp angle to each other. Envelopes of sporonts and sporoblasts possess typical 2-membrane structure. Their nuclei lay in pairs like in Nosema species. Sporoblasts (unlike meronts and sporonts) are surrounded by the electron lucid zone of the cytoplasm with numerous tubule-like and spherical inclusions. Host mitochondria are closely adjacent to the periphery of this zone. The formation of characteristic spherical structures (globules), in which the posterior ends of two (in rare cases of 1 or 4) sister spores are downsinked, starts immediately after the sporont division. Globules rapidly increase in size during the sporogenesis. The homogenous globule (6-8 microns), which is essentially larger than a spore, seems to represent the modified exospore; it is not limited by any visible membrane. Fine structure of spores (2.5-3.0 x 2.0-2.3 microns) is typical for the nosems: spherical polar disk; lamellas polaroplast; nuclei located along the long spore axis; basal part of a polar tube forming a fork. A thin (25-40 nm) heterophilar polar filament forms 12 rows of various structure (8 + 4) packed in two rows. The posterior vacuole was no revealed. Endospore is very thick up to 250-400 nm in mature spores. The exospore covering the 2/3-4/5 of the spore length passes into the globule on the posterior end. The refined diagnosis of Issia globulifera: type host species--Anopheles messae (larvae, pupae); tissue localization: mitgut epithelium; terra typica: Siberia, Tomsk region, Ob river basin; developmental stages: multicellular merogonial plasmodium, sporont gives rise to 2 (rarely 1 or 4) sporoblasts connected with each other by a "globule", a specific formation homologous to an exospore of other microsporidians; globule (6-8 microns) possesses a homogenous inner structure of average electron density; sporophorous vesicle not formed; spore structure is typical for Nosematidae. Embeddings, negatives (## 2301, 2303, 2308, 2311-2313, 2532, 2533) and photos of type material in the collection of All-Russian Institute for Plant Protection.

[Parasitophorous vacuole of microsporidia]. / O parazitofornoi vakuoli mikrosporidii.

The comparative analysis of the ultrastructure of various types of parasitophorous vacuoles (PV) induced by microsporidians is given. The data on the occurrence of PV in the hosts belonging to different systematic phyla are summarised. It is concluded, that the formation of PV around microsporidians might take place either in certain parasite species or in the special type of the invaded cells, or could be connected with the development in the unspecific host. The variety of fine structure of PV might be explained by an extremely broad range of hosts (from protists to mammalians), with different level of development of their immune system (defence reactions). Three basic types of PV are proposed according the organization of their envelopes (walls): (1) a single membrane originated from the host cell plasmalemma (hosts: Aves and Mammalia); (2) a single membrane derived from the host ER (hosts: Pisces); (3) a single- or double-membrane host ER (hosts: protists, invertebrates and animals of other systematic groups). It was assumed that the formation of the PV around microsporidians reflects more primitive host-parasite interactions, than the development of the parasite in a direct contact with the host cell cytoplasm.