Scanning electron microscopic observation of the in vitro cultured protozoan, Perkinsus olseni, isolated from the Manila clam, Ruditapes philippinarum.

BACKGROUND: Perkinsosis is a major disease affecting the commercially important marine mollusk Ruditapes philippinarum (Manila clam) in Asian waters. In this study, we investigated the morphological characteristics of Perkinsus olseni, the causative agent of perkinsosis, cultured under laboratory conditions at different stages of its life cycle using a scanning electron microscope (SEM). RESULTS: The prezoosporangia formed after induction with Ray's fluid thioglycollate medium (RFTM) developed into zoosporangia. During this process, a discharge tube formed a porous sponge-like structure that detached before the zoospores were released; thus, this organelle operated as a bung. Liberated zoospores gradually transformed into immature trophozoites, during which detachment of the anterior flagella occurred, but the loss of the posterior flagella was not clearly observed in the present study. Mature trophozoites underwent schizogony by cleaving the cell forming some merozoites in schizonts, which were released by the rupturing of the cellular membrane of the schizont within a few days. CONCLUSIONS: Our morphological and ultrastructural studies contribute new information on the life cycle and propagation of P. olseni.

Native parasite affecting an introduced host in aquaculture: cardiac henneguyosis in the red seabream Pagrus major Temminck & Schlegel (Perciformes: Sparidae) caused by Henneguya aegea n. sp. (Myxosporea: Myxobolidae).

BACKGROUND: Henneguya Thélohan, 1892 (Myxobolidae) is one of the most species-rich genera of myxosporean parasites infecting fish. Although common in nature, there are few reports of these parasites causing important disease in aquaculture. In this paper, we describe a new species of Henneguya infecting Pagrus major (Temminck & Schlegel), a fish host introduced to the Mediterranean Sea from Japan in the late 1980s. RESULTS: Large plasmodia of the parasite were found in the bulbus arteriosus and in the ventricle of the infected fish. Spores were found mainly in the kidney and heart and were accompanied by melanized macrophages or vascular intimal proliferation mixed with a mild non-suppurative response, respectively. Comparisons of morphometric data for spore and polar capsule length and width, suggest a unique combination of features in the newly described species. Molecular analysis, based on 18S rDNA sequence of the parasite, followed by phylogenetic analysis, indicated that the parasite described here is a novel species of Henneguya, clustered with the marine congeneric species. CONCLUSIONS: Henneguya aegea n. sp. infects in aquaculture P. major, a host introduced as eggs to the Mediterranean from Japan. Despite the high host specificity of the myxobolid parasites, H. aegea n. sp. seems to be able to use P. major as a host and propagate successfully, causing morbidity and mortality. This could result in spillback of the new species from high density cultured non-native P. major to native fish hosts.

Phylogeny of Physarida (Amoebozoa, Myxogastria) Based on the Small-Subunit Ribosomal RNA Gene, Redefinition of Physarum pusillum s. str. and Reinstatement of P. gravidum Morgan.

Myxomycetes (also called Myxogastria or colloquially, slime molds) are worldwide occurring soil amoeboflagellates. Among Amoebozoa, they have the notable characteristic to form, during their life cycle, macroscopic fruiting bodies, that will ultimately release spores. Some 1,000 species have been described, based on the macroscopic and microscopic characteristics of their fruiting bodies. We were interested in Physarum pusillum (Berk. & M.A. Curtis) G. Lister, a very common species described with two variants, each bearing such morphological differences that they could represent two distinct species. In order to test this, we observed key characters in a large selection of specimens attributed to P.  pusillum, to its synonyms (in particular Physarum gravidum), and to related species. In addition, the small-subunit ribosomal RNA gene was obtained from seven of these specimens. Based on these data, we provide a comprehensive phylogeny of the order Physarida (Eukaryota: Amoebozoa: Conosa: Macromycetozoa: Fuscisporidia). Morphology and phylogeny together support the reinstatement of P. gravidum Morgan 1896 with a neotype here designated, distinct from P. pusillum, here redefined.

Phylogeny of Myxobolidae (Myxozoa) and the evolution of myxospore appendages in the Myxobolus clade.

Genera Myxobolus Bütschli, 1882 and Henneguya Thélohan, 1892 (Myxobolidae) are specious myxozoan genera. They comprise nearly half of overall known myxozoan species diversity. A typical spore feature of Henneguya is the presence of two caudal appendages of the spore valves, which distinguishes them from species of the genus Myxobolus. Several Myxobolus spp., however, were reported to show aberrant spores with Henneguya-like caudal appendages. We found such aberrant spores in Myxobolus tsangwuensis and Myxobolus wulii. We studied the ultrastructure of M. wulii and Myxobolus oralis spores with caudal appendages by transmission electron microscopy (TEM). TEM of these aberrant spores revealed that their caudal appendages have the same ultrastructure as the appendages of Henneguya spp. Small caudal appendages of M. wulii spores observed only on TEM suggested that this character may be often overlooked and more Myxobolus species potentially have the ability to express the caudal appendages on the myxospore. In order to trace the evolution of this character, we performed broad phylogenetic analysis of all species of the family Myxobolidae which are available in GenBank including nearly 300 taxa. We found at least eight independent evolutionary origins of spores with two appendages, three origins of a single appendage and 12 apparent secondary losses of the spore projections. Therefore, genus Henneguya with typical two-tailed myxospores is polyphyletic, however a majority of its species has a common ancestor and groups in the second largest subclade of the Myxobolus clade. We also mapped the biological characteristics (host, site of infection and environment) of Myxobolidae species on the phylogenetic tree. We revealed an evident host-associated evolutionary pattern in all parts of the Myxobolus clade with a distinct and species-rich subclade containing almost exclusively species infecting species of the Order Cypriniformes.

First Myxozoan Infection (Cnidaria: Myxosporea) in a Marine Polychaete from North America and Erection of Actinospore Collective Group Saccimyxon.

In a survey of marine annelids for myxosporean infection in Charleston Harbor, South Carolina, we collected 3,214 polychaetes from 21 families and found infections in 6 spionid individuals. Based on gross morphology and COI sequencing, all infected spionids were identified as Streblospio benedicti. Infection prevalence was 0.8% (6/734) of that species of spionid, and 0.2% of all 3,214 polychaetes examined. Pansporocysts contained 8 actinospores and developed in the tegument of the annelid host. This is the first myxozoan infection recorded from this polychaete species, second in the family, and the first marine myxozoan found in the Americas. It is the first marine species found to develop in the tegument of its annelid host; a site of development observed only once before, in Ceratonova shasta infections of its freshwater sabellid polychaete host. Mature actinospores were morphologically simple, truncated ellipsoids, lacking processes or ornamentation, 9.0 ± 0.5 µm × 6.0 ± 0.4 µm. Their sack-like shape was similar to 9 of the 12 actinospores described previously from polychaetes; 10/12 had been and ascribed originally to the morphological collective group Tetractinomyxon despite 9 of these having few similarities to the original description of this group. We propose to name the simple, spherical to ellipsoidal spore morphotype Saccimyxon to encompass both our novel actinospore and the 9 other sack-like polychaete actinospore types in the literature. In the present study, 18S rDNA sequencing demonstrated that the myxozoans that infected the 6 spionids were genetically the same species (type sample 1,737 nucleotides, GenBank accession number MH791159) and was not >95% similar to any sequence in GenBank. Phylogenetic analysis showed that the myxozoan species we encountered is basal to the kudoids and thus likely to have a morphologically simple myxospore stage with fewer than 4 valves. However, without a genetic match, the presumptive vertebrate host remains unknown.

Supplementary studies on Henneguya guanduensis (Cnidaria: Myxosporea) infecting gills and intestine of Hoplosternum littorale in Brazil: Ultrastructural and molecular data.

Henneguya guanduensis was originally described as a gill parasite of the Hoplosternum litoralle, a siluriform fish belonging to the Callichthyidae family. A study was undertaken with 58 specimens of H. litoralle taken from Batalha River in the state of São Paulo. The fish were collected and examined searching for lesions and/or myxosporean plasmodia. The prevalence of infection was 9.31% in the gills and 5.17% in the intestine. The mature spores had elongated bodies with polar capsules of inequal size and a caudal length greater than body length. Morphological characteristics identified the parasite as H. guanduensis. Molecular analysis of the SSU rDNA partials sequences resulted in a 1796 bp and 1712 bp for gills and intestine repectively, demonstrating significant genetic differences with previously described species of Henneguya and 99.7% similarity to each other when aligned. Phylogenetic analysis comparing the SSU rDNA sequence of H. guanduensis with closest species as indicated by BLASTn Max Score showed H. guanduensis as sister species of H. loretoensis in a subclade compose by species that parasites fishes from Amazon basin. This is the first report of the finding of Henneguya guanduensis spores in the intestine of Hoplosternum littorale.

Ultrastructure of vegetative cells and resting cysts, and live observations of the encystation and excystation processes in Diophrys oligothrix Borror, 1965 (Protista, Ciliophora).

Ciliated protists can form cysts to resist unfavorable environmental conditions and then excyst when environmental conditions become favorable. This study used electron and light microscopy to investigate the structure of vegetative cells and resting cysts, as well as the encysting and excysting processes of Diophrys oligothrix. For the first time, ampules were revealed beneath the pellicle in the genus Diophrys, and their extrusome types differed between Diophrys species. Membrane-packed discs of diverse shapes were found in the cytoplasm just beneath the pellicle around the cytopharynx and were separated by rows of microtubule units. Beneath the discs, some double-layer microtubule structures were detected as well. During encystment, the ventral ciliature was folded in a ventral cavity of the cell, and the caudal cirri were retracted directly into the cyst in a separate cavity on the dorsal side. In the resting cysts, high autophagic activity occurred, possibly including digestion of membrane-packed discs and ampules. Two macronuclear nodules kept their basic shape, although the chromatin aggregation and fusion region were observed in ultrathin sections. The cyst wall contained two layers, namely, the ectocyst, and endocyst. In mature cysts, basal bodies and ciliary shafts were observed, demonstrating that D. oligothrix forms non-kinetosome-resorbing cysts. The process of excystment occurred in two modes, either with or without participation of a contractile vacuole.

Morphology and phylogeny of Ameson portunus n. sp. (Microsporidia) infecting the swimming crab Portunus trituberculatus from China.

Ameson portunus n. sp. is a new microsporidian species that infects the skeletal muscle of Portunus trituberculatus, a pond-reared swimming crab from China. This parasite was characterized using morphological and molecular phylogenetic data. Light and transmission electron microscopy revealed that this microsporidian experienced disporogonic and polysporogonic (chain-like) life cycles. Mature uninucleate spores appeared ovoid, measured 1.4±0.06×1.0±0.07µm on ultrathin sections, and exhibited no dimorphism. The isofilar polar filament was coiled in 8-9 turns. Of these coils, 5-9 were arranged in large regular outer layers; the remaining coils (0-3 coils) were situated internally. According to phylogenetic analyses based on the small subunit (SSU) rDNA gene, A. portunus n. sp. belonged to the group comprising Ameson spp. and Nadelspora canceri. The result of comprehensive analysis of ultrastructural features, molecular phylogenetic data, host and geographical differences among known species supports the establishment of a new Ameson species for this parasite. Ameson portunus n. sp. is the first Ameson species described from the coasts of East Asia.

Redescription of Chloromyxum ellipticum Li & Nie, 1973 (Myxosporea: Chloromyxidae) infecting the gall bladder of grass carp Ctenopharyngodon idellus Valenciennes, 1844, supplemented by morphological and molecular characteristics.

The traditional taxonomy of the genus Chloromyxum Mingazzini, 1890 has been intensively challenged to be paraphyletic by recent ribosomal DNA (rDNA)-based phylogenetic analysis. Undersampling to get rich sequence data to infer more scientific phylogenetic relationships makes scientists conservatively assign all non-marine elasmobranch-infecting species as Chloromyxum sensu lato. Although complex ridge pattern on the spore surface observed by scanning electron microscopy was thought to be critical for the identification of Chloromyxum species, insufficient data also prevent this ultrastructural data to be a valid taxonomic feature for this genus. It is especial for Chloromyxum species to be reported in China. Molecular and ultrastructural characteristics are yet available for all 22 Chloromyxum species recorded in China. During the investigation of the diversity of coelozoic fish myxosporeans, Chloromyxum ellipticum Li & Nie, 1973 was found to highly infect the gall bladder of Ctenopharyngodon idellus Valenciennes, 1844 in Poyang Lake watershed of Jiangxi province, Eastern China. Here, we redescribed it by the currently recommended holistic approach of combining morphological, ultrastructural, and molecular characteristics. Mature spores were found floating free in the gall bladder, but no plasmodium observed. Spores are typical freshwater teleost-infecting Chloromyxum species, spherical or subspherical in lateral view, measuring 7.7 ± 0.08 µm (6.9-9.1) in length, 6.3 ± 0.09 µm (5.6-7.6) in width, and 5.8 ± 0.20 µm (5.2-6.3) in thickness. Four pyriform polar capsules, located at the anterior end of the spores, were equal in size, 3.3 ± 0.06 µm (2.2-4.1) long and 2.1 ± 0.03 µm (1.7-2.5) wide. Polar filaments coiled with four to five turns. Two equal spore valves are symmetrical, with 10-16 surface extrasutural ridges per valve, aligned along the longitudinal axis. The obtained partial 18S rDNA of C. ellipticum did not match any sequences available in GenBank. Phylogenetic analysis showed that C. ellipticum clustered firstly with Chloromyxum legeri with robust nodal support and grouped then with urinary system of freshwater teleost-infecting Chloromyxum clade, rather than other gall bladder of freshwater teleost-infecting clade.

Host Diet Affects the Morphology of Monarch Butterfly Parasites.

Understanding host-parasite interactions is essential for ecological research, wildlife conservation, and health management. While most studies focus on numerical traits of parasite groups, such as changes in parasite load, less focus is placed on the traits of individual parasites such as parasite size and shape (parasite morphology). Parasite morphology has significant effects on parasite fitness such as initial colonization of hosts, avoidance of host immune defenses, and the availability of resources for parasite replication. As such, understanding factors that affect parasite morphology is important in predicting the consequences of host-parasite interactions. Here, we studied how host diet affected the spore morphology of a protozoan parasite ( Ophryocystis elektroscirrha ), a specialist parasite of the monarch butterfly ( Danaus plexippus ). We found that different host plant species (milkweeds; Asclepias spp.) significantly affected parasite spore size. Previous studies have found that cardenolides, secondary chemicals in host plants of monarchs, can reduce parasite loads and increase the lifespan of infected butterflies. Adding to this benefit of high cardenolide milkweeds, we found that infected monarchs reared on milkweeds of higher cardenolide concentrations yielded smaller parasites, a potentially hidden characteristic of cardenolides that may have important implications for monarch-parasite interactions.

Myxobolus chushi n. sp. (Myxozoa:Myxosporea) parasitizing Schizothorax niger (Heckel), a native cyprinid fish from Wullar Lake in Kashmir Himalayas.

In the study, a new species, Myxobolus chushi n. sp. infecting gills of wild specimens of Schizothorax niger (Heckel) inhabiting Wullar Lake in Kashmir Himalayas, (J&K) India has been described based on morphology of the myxospore and using partial 18S rDNA sequencing. Pathological changes in the gills have been studied with the help of histological sections stained with Luna's method. Twenty fish specimens were examined, out of which four had oval, white plasmodia in gills measuring 2.0×0.5mm. The myxospores were spherical to ovoidal in shape with slightly attenuated posterior end, measuring 11.17±0.23 (10.60-11.40)µm in length and 9.14±0.06 (8.80-9.20)µm in width, having a prominent pore at the anterior end. The polar capsules were pyriform in shape, measuring 4.25±0.15 (4.00-4.40)µm in length and 2.38±0.27 (2.00-2.65)µm in width having polar filaments forming coils up to 5 in number. Parietal folds 9 in number present on the posterior part of the shell. The intensity of infection was recorded to be moderate as indicated by gill plasmodial index (GPI=2). The plasmodium was located in the vascular network occupying whole of the gill lamella therefore typed as intralamellar vascular type, LV3. Analysis of 18S small subunit (SSU) rDNA sequence of the isolate demonstrated 90% homogeneity with M. sp. KLT-2014 infecting scales of Labeo rohita from Myanmar and 89% with M. dermiscalis infecting scales of Labeo rohita from India.

Ultrastructural description of Myxobolus cuneus (Myxosporea) in the skeletal muscle and kidney of tropical farmed fish Piaractus mesopotamicus (Characiformes: Characidae).

This study characterizes by transmission electron microscopy (TEM) and morphometric features the myxozoan Myxobolus cuneus (Myxosporea) in Piaractus mesopotamicus and reports the skeletal muscle and kidney as site of infection. The register was based in 21 young fish from intensive fish farming in Southeast Brazil and the spores were analyzed in fresh-mounted slides of the infected organs stained with Toluidine blue and processed as usual for TEM. It differs from Myxobolus cunhai from the fish host and different polar capsule size, and from Myxobolus serrasalmi on the pyriform spore shape and an oval macrospore, differently to that reported in this study. Morphometric characteristics and TEM study confirmed the present material as M. cuneus.

A novel paramyxean parasite, Marteilia octospora n. sp. (Cercozoa) infecting the Grooved Razor Shell clam Solen marginatus from Galicia (NW Spain).

Protozoan parasites of the genus Marteilia have been detected in marine bivalves and other invertebrates around the world, associated in some cases with mass mortalities. The present paper reports the characterization of the Marteilia sp. protozoan infecting the digestive gland of the Grooved Razor Shell clam (Solen marginatus) from Galicia (NW Spain), proposing a novel species in the genus: Marteilia octospora n. sp. Morphological and molecular techniques were used for the description of this parasite. Tissue imprints were essential in the study to confirm the presence of 8 spores per sporangium, a number never reported in other species from this genus. An ultrastructural study revealed that the size and number of dense granules, free in the mature sporangia, were quite different from granules in other Marteilia spp. Another morphological difference is the absence of a layer of concentric membranes found surrounding the mature spore in other species. In addition, concentric membranous structures observed in the different stages of the parasite have never been mentioned in other species of genus Marteilia. Moreover, molecular analysis of the rDNA intergenic spacer (IGS) and the internal transcribed spacer (ITS-1) showed differences with the sequences available for other Marteilia spp.

Characterisation of 20S Proteasome in Tritrichomonas foetus and Its Role during the Cell Cycle and Transformation into Endoflagellar Form.

Proteasomes are intracellular complexes that control selective protein degradation in organisms ranging from Archaea to higher eukaryotes. These structures have multiple proteolytic activities that are required for cell differentiation, replication and maintaining cellular homeostasis. Here, we document the presence of the 20S proteasome in the protist parasite Tritrichomonas foetus. Complementary techniques, such as a combination of whole genome sequencing technologies, bioinformatics algorithms, cell fractionation and biochemistry and microscopy approaches were used to characterise the 20S proteasome of T. foetus. The 14 homologues of the typical eukaryotic proteasome subunits were identified in the T. foetus genome. Alignment analyses showed that the main regulatory and catalytic domains of the proteasome were conserved in the predicted amino acid sequences from T. foetus-proteasome subunits. Immunofluorescence assays using an anti-proteasome antibody revealed a labelling distributed throughout the cytosol as punctate cytoplasmic structures and in the perinuclear region. Electron microscopy of a T. foetus-proteasome-enriched fraction confirmed the presence of particles that resembled the typical eukaryotic 20S proteasome. Fluorogenic assays using specific peptidyl substrates detected presence of the three typical peptidase activities of eukaryotic proteasomes in T. foetus. As expected, these peptidase activities were inhibited by lactacystin, a well-known specific proteasome inhibitor, and were not affected by inhibitors of serine or cysteine proteases. During the transformation of T. foetus to endoflagellar form (EFF), also known as pseudocyst, we observed correlations between the EFF formation rates, increases in the proteasome activities and reduced levels of ubiquitin-protein conjugates. The growth, cell cycle and EFF transformation of T. foetus were inhibited after treatment with lactacystin in a dose-dependent manner. Lactacystin treatment also resulted in an accumulation of ubiquitinated proteins and caused increase in the amount of endoplasmic reticulum membranes in the parasite. Taken together, our results suggest that the ubiquitin-proteasome pathway is required for cell cycle and EFF transformation in T. foetus.

Twelve myxosporean species of the family Myxobolidae infecting freshwater fishes of the River Nile, Egypt, with the description of four novel species.

Myxosporidian parasites infecting fish are very dangerous parasites causing severe damage to a large number of economically important fishes especially in aquaculture. A survey of myxosporean parasites infecting four species of fishes from the River Nile in Egypt is conducted. One hundred and ninety-five out of 316 fish specimens with a percentage of 61.7% were found to be naturally infected with these parasites. Light microscopic examination of different tissues revealed the presence of 12 myxosporean species belonging to the family Myxobolidae. Four of the identified species are novel and the other eight species are redescribed. Myxidium sp.nov. a coelozoic species inhabiting the gallbladder of Labeo niloticus with its mature spores float free in bile was detected. These spores possess a fusiform, straight, or slightly crescentic shape with less pointed ends and two equal polar capsules. Three novel histozoic Myxobolus species infecting Oreochromis niloticus were identified. Myxobolus sp(1).nov. is a species inhabiting kidney tissue with ovoid spores exhibiting a small intercapsular appendix. Myxobolus sp(2).nov. and Myxobolus sp(3).nov. recovered from kidney and intestinal tissues. Spores of Myxobolus sp(2).nov. are elliptical in shape with an anterior end wider than posterior one. Their two polar capsules are ovoid to pyriform occupied nearly the first third of the spore body. Spores of Myxobolus sp(3).nov. are broader than long with nearly rounded or ovoid two polar capsules. Eight species of the recovered myxosporean parasites are redescribed, Myxobolus niloticus Fahmy et al., 1971 from pectoral, dorsal, and tail fins of L. niloticus, Henneguya suprabranchiae Landsberg, 1987, and Henneguya branchialis Ashmawy et al., 1989 are recovered from the gills and suprabranchial organ of the catfish Clarias gariepinus, respectively, Myxobolus naffari Abdel-Ghaffar et al., 1998 and Myxobolus imami Ali et al., 2002 are found in the kidney of Barbus bynni and L. niloticus, Myxobolus caudatus Ali et al. & Parasitol Res (2002) from Tail fin of B. bynni, Myxobolus fomenai Abdel-Ghaffar et al., 2008 from kidney and intestinal tissues of O. niloticus, Thelohanellus niloticus Abdel-Ghaffar et al., 2012 are observed in the gills of L. niloticus.

Nivicolous Stemonitales from the Austral Andes: analysis of morphological variability, distribution and phenology as a first step toward testing the large-scale coherence of species and biogeographical properties.

Nivicolous myxomycetes occur at the edge of spring-melting snow in mountainous areas. They are mostly considered cosmopolitan species morphologically and ecologically uniform across their entire distribution ranges. Thus, long-distance dispersal has been suggested to be the main mechanism shaping their ranges and geographical variability patterns. To test this hypothesis we conducted the first detailed analysis of morphological variability, occurrence frequency and phenology of nivicolous myxomycetes collected in the hitherto unexplored Austral Andes of South America (southern hemisphere = SH) in the comparative context of data from the northern hemisphere (NH). We used Stemonitales, the most representative and numerous taxonomic order in nivicolous myxomycetes, as a model. A total of 131 South American collections represented 13 species or morphotypes. One of them, Lamproderma andinum, is new to science and described here. Several others, L. aeneum, L. album, L. pulveratum, "Meriderma aff. aggregatum ad. int.", M. carestiae and "M. spinulosporum ad. int.", were previously unknown from the SH. Lamproderma ovoideum is reported for the first time from South America and Collaria nigricapillitia is new for Argentina. The fine-scale morphological analysis of all species from the study area and reference NH material demonstrated a high intraspecific variability in most of them. This suggests isolation and independent evolutionary processes among remote populations. On the other hand, the uniform morphology of a few species indicates that long-distance dispersal is also an effective mechanism, although not as universal as usually assumed, in some nivicolous myxomycetes. Analysis of nivicolous species assemblages also showed significant differences among major geographic regions in that the Stemonitales were significantly less common in the SH than in the NH. Furthermore, the occurrence of nivicolous species in summer and autumn, out of the typical phenological season, is recognized as a possible distinctive phenomenon for the SH populations.

In vitro cultivation of Hematodinium sp. isolated from Atlantic snow crab, Chionoecetes opilio: partial characterization of late developmental stages.

Hematodinium is a parasitic dinoflagellate of numerous crustacean species, including the economically important Atlantic snow crab, Chionoecetes opilio. The parasite was cultured in vitro in modified Nephrops medium at 0 °C and a partial characterization of the life stages was accomplished using light and transmission electron microscopy (TEM). In haemolymph from heavily infected snow crabs two life stages were detected; amoeboid trophonts and sporonts. During in vitro cultivation, several Hematodinium sp. life stages were observed: trophonts, clump colonies, sporonts, arachnoid sporonts, sporoblasts and dinospores. Cultures initiated with sporonts progressed to motile dinospores; however, those initiated with amoeboid trophonts proliferated, but did not progress or formed schizont-like stages which were senescent artefacts. Plasmodial stages were associated with both trophonts and sporonts and could be differentiated by the presence of trichocysts on TEM. Macrodinospores were observed but not microdinospores; likely due to the low number of Hematodinium sp. cultures that progressed to the dinospore stage. No early life stages including motile filamentous trophonts or gorgonlocks were observed as previously noted in Hematodinium spp. from other crustacean hosts. All Hematodinium sp. life stages contained autofluorescent, membrane-bound electron dense granules that appeared to degranulate or be expelled from the cell during in vitro cultivation.

[THE DESCRIPTION OF MYXOBOLUS PELECICOLA SP. NOV. (MYXOZOA: MYXOBOLIDAE) FROM PELECUS CULTRATUS (CYPRINIFORMES, CYPRINIDAE)].

Morphological study of myxosporean parasites Myxobolus pelecicola sp. n. from Pele- cus cultratus (L., 1758), is presented. Presporogenic stage not observed. Polysporous, el- lipsoidal. plasmodia up to 1.2 mm in length and 0.1 to 0.4 mm in width develop in the ske- letal musculature. Mature spores elongate oval or oval form in frontal view, lens-shaped in sutural view. Length of spores 16.5 ± 0.83 (14.9-18.0), width 12.1 ± 0.63 (11.6-13.3) (n = 50), thickness 7.8 ± 0.3 (7.4-8.3) µm (n = 10). Polar capsules pyriform, unequal, length of large 8.0 ± 0.44 (7.5-8.8), small 7.6 ± 0.26 (7.2-7.8), width of large 4.5 ± 0.22 (4.2-4.7), small 4.0 ± 0.21 (3.8-4.3) µm. Five to 6 distinct filament coils oriented obli- quely to capsule length, found in polar capsule. The length of extruded (in water) filament up to 130 µm. Large, but indistinct triangular intercapsular appendix present in spores. Single rodlet inclusion is very often observed in the posterior part of spore. Mucous enve- lope is absent. The type host of Myxobolus pelecicola sp. n. is sabrefish Pelecus cultratus (L., 1758). Type locality: Finnish Bay (60°05' N 29°55' E) and Ladoga lake (60°07' N 32°19' E), Russia.

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.

Using environmental niche models to test the 'everything is everywhere' hypothesis for Badhamia.

It is often discussed whether the biogeography of free-living protists is better explained by the 'everything is everywhere'(EiE) hypothesis, which postulates that only ecology drives their distribution, or by the alternative hypothesis of 'moderate endemicity' in which geographic barriers can limit their dispersal. To formally test this, it would be necessary not only to find organisms restricted to a geographical area but also to check for their presence in any other place with a similar ecology. We propose the use of environmental niche models to generate and test null EiE distributions. Here we have analysed the distribution of 18S rDNA variants (ribotypes) of the myxomycete Badhamia melanospora (belonging to the protozoan phylum Amoebozoa) using 125 specimens from 91 localities. Two geographically structured groups of ribotypes congruent with slight morphological differences in the spores can be distinguished. One group comprises all populations from Argentina and Chile, and the other is formed by populations from North America together with human-introduced populations from other parts of the world. Environmental climatic niche models constructed separately for the two groups have significant differences, but show several overlapping areas. However, only specimens from one group were found in an intensively surveyed area in South America where both niche models overlap. It can be concluded that everything is not everywhere for B. melanospora. This taxon constitutes a complex formed by at least two cryptic species that probably diverged allopatrically in North and South America.