Microsporidian xenomas in fish seen in wider perspective

The history of understanding xenoparasitic complexes or xenomas provoked in the host cell by various protists and especially by microsporidia is outlined. Microsporidia have been known to produce xenomas in oligochaetes (e.g., genera Bacillidium, Burkea, Hrabyeia, Jirovecia, species of the collective group Microsporidium), crustaceans (e.g., Abelspora, Mrazekia), insects (e.g., Polydispyrenia, Thelohania) and poikilothermic vertebrates, mostly fish (Alloglugea, Amazonspora, Glugea, Ichthyosporidium, Loma, Microfilum, Microgemma, Neonosemoides, Pseudoloma, Spraguea, Tetramicra). An overview of characters of xenomas caused by species of these genera is presented. The study of microsporidia causing xenomas in fish offers an insight into cell pathology and is of interest since many of these species are important agents of diseases in commercial fish. Xenomas produced from a few types of target cell display a complete change of organisation of the host cell and differ, according to the agent, in their structure. Recent data show that proliferation of the parasite may have already started in the cells transporting the parasites to the final site of xenoma formation. However, these are preliminary revelations and most of the facets of the life cycle are still to be clarified. Curiously, xenoma-forming microsporidia do not seem to be strictly host specific. The salient features of fish microsporidian xenomas are discussed, such as role of the xenoma, whether its features are host- or microsporidium-dependent, development and demise of the xenoma in the course of time, and host reaction phenomena. The need of further research is emphasised.     

Microgemma vivaresi (Microsporidia: Tetramicridae): host reaction to xenomas induced in sea scorpions, Taurulus bubalis (Osteichthyes: Cottidae)

Xenomas caused by Microgemma vivaresi Canning, Feist, Longshaw, Okamura, Anderson, Tsuey Tse et Curry, 2005 were found in liver and skeletal muscle of sea scorpions, Taurulus bubalis (Euphrasen). All muscle xenomas examined were in an advanced stage of destruction. In developing xenomas found in liver, parasites were restricted to the centre of the cell, separated from a parasite-free zone by a nuclear network formed by branching of the host cell nucleus. Although xenomas were able to reach a size of several hundred microns, the surface remained a simple plasma membrane. Host reactions took the form of penetration by phagocytes and isolation by fibroblasts. Once the xenoma had been attacked, the nuclear profiles became pycnotic and the barrier between parasitized and parasite-free zones was lost. Parasite antigens cannot be exposed at the surface of intact xenomas, as the host does not recognise the enlarging cell as foreign. Breaches in the plasma membrane of the xenoma and leakage of parasite antigens are thought to be the stimuli for phagocyte entry into the cell, its isolation by fibroblasts and eventual granuloma formation.     

Myxobolus intrachondrealis sp. n. (Myxosporea: Myxobolidae), a parasite of the gill cartilage of the common carp, Cyprinus carpio

A species not identifiable with any of the about 23 Myxobolus species recorded from the common carp so far, was detected in the gills of one- and two-summer-old specimens of the common carp ((Cyprinus carpio L.) cultured in pond farms in Hungary. The strictly tissue-specific plasmodia of the parasite were located, surrounded by hyaline cartilage cells, in the chondrous substance of the terminal parts of the gill arches and in the cartilage structure ventrally connecting the gill arches. The spores of the parasite described as Myxobolus intrachondrealis sp. n. developed in globular or ellipsoidal plasmodia measuring 300-600 microm. By their elongated ellipsoidal shape and similarly elongated polar capsules the spores were well distinguishable from the hitherto described Myxobolus species parasitic in the common carp and also from the cartilage-parasitic Myxobolus species of other fishes.     

Studies on transmission and life cycle of Enteromyxum scophthalmi (Myxozoa), an enteric parasite of turbot Scophthalmus maximus

In order to elucidate the transmission and dispersion routes used by the myxozoan parasite Enteromyxum scophthalmi Palenzuela, Redondo et Alvarez-Pellitero, 2002 within its host (Scophihalmus maximus L.), a detailed study of the course of natural and experimental infections was carried out. Purified stages obtained from infected fish were also used in in vitro assays with explants of uninfected intestinal epithelium. The parasites can contact and penetrate loci in the intestinal epithelium very quickly. From there, they proliferate and spread to the rest of the digestive system, generally in an antero-posterior pattern. The dispersion routes include both the detachment of epithelium containing proliferative stages to the intestinal lumen and the breaching of the subepithelial connective system and local capillary networks. The former mechanism is also responsible for the release of viable proliferative stages to the water, where they can reach new fish hosts. The finding of parasite stages in blood smears, haematopoietic organs, muscular tissue, heart and, less frequently, skin and gills, suggests the existence of additional infection routes in transmission, especially in spontaneous infections, and indicates the role of vascular system in parasite dispersion within the fish. The very high virulence of this species in turbot and the rare development of mature spores in this fish may suggest it is an accidental host for this parasite. This may also question the existence of a two-host life cycle involving an actinosporean stage in this species. Further studies are needed to clarify this open point of the life cycle.     

Trickle and single infection with Discocotyle sagittata (Monogenea: Polyopisthocotylea): effect of exposure mode on parasite abundance and development

Experimental infection of rainbow trout Oncorhynchus mykiss (Walbaum) with the monogenean Discocotyle sagittata (Leuckart, 1842) allowed comparison between trickle and single exposure, two infection modes demonstrated to occur in the wild. Both types of infection resulted in mean larval attachment success around 50%, which was significantly dependent on dose of infective larvae used (P < 0.0001), but was not affected by mode of infection (P = 0.244). Worms recovered from fish exposed to the same number of oncomiracidia but different mode of infection differed in their rate of development. The developmental stage attained by parasites was significantly affected by number of infective larvae used (P = 0.005), and by the interaction between dose and mode of infection (P = 0.026), suggesting competition among attached larvae. Statistical analysis demonstrated that in the early stages of infestation, worm distribution over the gill arches can be explained by the relative amount of water flowing over them. One, two and three months post-infection parasite numbers were comparable (P = 0.805), but their observed distribution gradually decreased in gill arches III and IV and increased in gill arch I, suggesting that parasites migrate after initial attachment. These results reproduce phenomena observed in the field, indicating that the experimental infection system could be employed to study infection dynamics and host-parasite interactions under controlled conditions.     

Proliferative renal myxosporidiosis in spawning coho salmon (Oncorhynchus kisutch) in British Columbia and Washington

An unidentified myxosporean parasite (CKX) is described from the kidney of approximately 80% of spawning coho salmon Oncorhynchus kisutch (Walbaum) in British Columbia, Canada and Washington, United States of America. Morphological features were described using light and electron microscopy. Sequencing of polymerase chain reaction (PCR) amplified 18S ribosomal RNA gene and in situ hybridisation were used to further characterise CKX. The parasite occurred with a focal distribution within tubule epithelial cells, the tubule lumen and the interstitium as primary cells containing from one to at least 16 secondary cells. Luminal stages were degenerate and sporogony was not observed. In situ hybridisation using a digoxygenin-labelled DNA probe confirmed CKX to be the source of DNA used in PCR analyses. CKX 18S rDNA sequences were most similar (97%) to those of Sphaerospora oncorhynchi. Phylogenetic analysis revealed similarities among the 18S rDNA sequences of CKX, S. oncorhynchi and Myxidium lieberkuehni. CKX is hypothesised to be the abortive extrasporogonic development of a Sphaerospora sp. or Myxidium sp. occurring in immune-incompetent spawning and senescent salmon.     

Sphaerospora elwhaiensis sp.n. (Myxosporea: Sphaerosporidae) from landlocked sockeye salmon Oncorhynchus nerka (Salmoniformes: Salmonidae) in Washington State, USA

A new species of sphaerosporid myxosporean, Sphaerospora elwhaiensis sp. n., is described from kidney of non-anadromous sockeye salmon (kokanee) Oncorhynchus nerka (Walbaum) from Lake Sutherland in the northern Olympic Peninsula, Washington, USA. Infection with the parasite was detected in 45% of 177 kokanee examined over 5 years. While conforming to the morphological criteria by which members of the genus are defined, the parasite is distinguished from congeners in salmonids of western North America by a unique combination of valvular sculpting of the myxospore, the relatively large size of the myxospore and monosporous development within the pseudoplasmodium. In addition, nucleotide sequences of the parasite's small and large subunit ribosomal RNA gene are unique. Phylogenetic analyses of these sequences suggested that the parasite is most closely related to freshwater Myxidium spp. and Zschokkella spp. The molecular data have provided further evidence for a polyphyletic association previously recognized among members of the genus and emphasize the need for a taxonomic revision of Sphaerospora Thélohan, 1892 and related genera.     

Homing of Gyrodactylus salaris and G. derjavini (Monogenea) on different hosts and response post-attachment

In natural European waters, the congeneric monogeneans Gyrodactylus derjavini Mikailov, 1975 and G. salaris Malmberg, 1957 are primarily found on brown trout Salmo trutta L. and Atlantic salmon Salmo salar L., respectively. Interestingly, rainbow trout, Oncorhynchus mykiss (Walbaum), originating from North America, is as susceptible as brown trout to G. derjavini. However, the mechanisms involved in this host specificity are poorly understood but may include behavioural, mechanical and chemical factors affecting parasite attraction, attachment, feeding, reproduction and host responses. In the present laboratory work, this question has been studied. Detached parasites (either G. derjavini or G. salaris) were offered a choice in small aquaria between fry of rainbow trout, Atlantic salmon and carp Cyprinus carpio L. Within 48 hours more than 90% of G. derjavini colonised rainbow trout and left salmon almost uninfected. Some parasites were found on carp. During the same time span, more than 60% of G. salaris attached to salmon, the rest infected rainbow trout and none were found on carp. Following attachment, the parasites need appropriate stimuli to initiate feeding and reproduction but even such a successful specific colonisation can be followed by a host response. Both humoral and cellular elements have been suggested to participate in these reactions but in the present work it was demonstrated by immunoblotting and immunocytochemistry that no antibodies in host mucus and host plasma bound to any parasite structures or epitopes.     

Fine structure of Henneguya pilosa sp. n. (Myxozoa: Myxosporea), parasite of Serrasalmus altuvei (Characidae), in Brazil

Henneguya pilosa sp. n., a new species of myxosporean from the gill filaments of the white piranha, Serrasalmus altuvei Ramirez, 1965 (Characidae), a freshwater teleost fish collected in the Zoological Garden of the city of Teresina (Piauí), Brazil, is described from light and transmission electron microscope observations. This myxosporean produced small plasmodia (up to 0.2 mm in diameter), each one containing all life-cycle stages of the parasite, including numerous spores. The spores, laterally compressed, averaged 54.2 (52.3-56.0) microm in total length and consisted of two unequal valves adhering together along the suture line and two caudal processes. The spore body measured 21.1 (20.0-23.1) microm in length, 5.9 (5.5-6.3) microm in width, and 2.2 (1.9-2.6) microm in thickness. The two equal ellipsoidal polar capsules of 7.4 (7.1-7.6) microm long and 1.2 (1.0-1.3) microm wide possessed a polar filament with 11-12 (rarely 13) turns. All surfaces of the spores were covered with a tightly adherent complex network of numerous densely ramified granulo-fibrillar masses, the longest measuring 1.5 microm long, observed around the caudal processes. The prevalence of infection was 30%. The taxonomic affinities of this parasite with other of the same genus in freshwater South American fish species are discussed.     

The development of Myxobolus pavlovskii (Myxozoa: Myxobolidae) includes an echinactinomyxon-type actinospore

Echinactinomyxon-type actinospores were found in a mixed-species oligochaete culture originating from the Temperate Water Fish Hatchery near Budapest, Hungary. On the basis of DNA sequence analysis, the actinospores were identified as Myxobolus pavlovskii (Akhmerov, 1954), the 18S rDNA sequence from myxospores of which is available in GenBank. Silver carp Hypophthalmichthys molitrix (Valenciennes) fry specimens were successfully infected by cohabitation with the echinactinomyxon-releasing oligochaetes, which confirmed the molecular data congruence. The echinactinomyxons and the myxospores that developed in the gills of exposed fish fry were analysed morphologically and on DNA basis. The infected gill tissue was examined histologically. As typical characters of M. pavlovskii, numerous small plasmodia were observed in the epithelia of gill lamellae. Plasmodia contained thousands of myxospores with polar capsules unequal in size and witl; large intercapsular processes. The 18S rDNA sequence from actinospores and those from myxospores originating from the experimentally infected fish were identical. The oligochaete species releasing actinospores was morphologically determined as Limnodrilus sp. This is the first record of an echinactinomyxon as an alternate stage within the genus Myxobolus.     

Distribution of Hatschekia pagellibogneravei (Copepoda: Hatschekiidae) on the gills of Pagellus bogaraveo (Teleostei: Sparidae) from Madeira, Portugal

A population of the gill parasite Hatschekia pagellibogneravei (Hesse, 1878) was studied on one of its sparid fish hosts, the blackspot seabream, Pagellus bogaraveo (Brünnich), off the coast of Madeira Island, Portugal, northeast Atlantic. Very high infection levels of this copepod were detected, with no significant seasonal differences. Abundance was negatively correlated with fish size. There were significant differences in the distribution of this copepod among the gill arches of the host, which seem to be best explained by differences in water flow within the gill habitat.     

The early events of Brachiola algerae (Microsporidia) infection: spore germination, sporoplasm structure, and development within host cells

Brachiola algerae (Vavra et Undeen, 1970) Lowman, Takvorian et Cali, 2000, originally isolated from a mosquito, has been maintained in rabbit kidney cells at 29 degrees C in our laboratory. This culture system has made it possible to study detailed aspects of its development, including spore activation, polar tube extrusion, and the transfer of the infective sporoplasm. Employing techniques to ultrastructurally process and observe parasite activity in situ without disturbance of the cultures has provided details of the early developmental activities of B. algerae during timed intervals ranging from 5 min to 48 h. Activated and nonactivated spores could be differentiated by morphological changes including the position and arrangement of the polar filament and its internal structure. The majority of spores extruded polar tubes and associated sporoplasms within 5 min post inoculation (p.i.). The multilayered interlaced network (MIN) was present in extracellular sporoplasms and appeared morphologically similar to those observed in germination buffer. Sporoplasms, observed inside host cells were ovoid, contained diplokaryotic nuclei, vesicles reminiscent of the MIN remnants, and their plasmalemma was already electron-dense with the "blister-like" structures, typical of B. algerae. By 15 min p.i., the first indication of parasite cell commitment to division was the presence of chromatin condensation within the diplokaryotic nuclei, cytoplasmic vesicular remnants of the MIN were still present in some parasites, and early signs of appendage formation were present. At 30 min p.i., cell division was observed, appendages became more apparent, and some MIN remnants were still present. By two hours p.i., the appendages became more elaborate and branching, and often connected parasite cells to each other. In addition to multiplication of the organisms, changes in parasite morphology from small oval cells to larger elongated "more typical" parasite cells were observed from 5 h through 36 h p.i. Multiplication of proliferative organisms continued and sporogony was well underway by 48 h p.i., producing sporonts and sporoblasts, but not spores. The observation of early or new infections in cell cultures 12-48 h p.i., suggests that there may also exist a population of spores that do not immediately discharge, but remain viable for some period of time. In addition, phagocytized spores were observed with extruded polar tubes in both the host cytoplasm and the extracellular space, suggesting another means of sporoplasm survival. Finally, extracellular discharged sporoplasms tightly abutted to the host plasmalemma, appeared to be in the process of being incorporated into the host cytoplasm by phagocytosis and/or endocytosis. These observations support the possibility of additional methods of microsporidian entry into host cells and will be discussed.     

Electron microscopic study of a new microsporean Microsporidium epithelialis sp. n. infecting Tubifex sp. (Oligochaeta)

The cytology of a new microsporean parasite Microsporidium epithelialis sp. n. from the intestinal epithelial cells of the freshwater oligochaete Tubifex sp. (Tubificidae) is described. The microsporean occurred together with an actinosporean of the genus Triactinomyxon, which was found between the epithelial cells. The merogonic and sporogonic stages (mature spores included) of the microsporean parasite are monokaryotic. An individual sporophorous vesicle surrounds each spore. The fixed and stained spore has an average dimension of 1.9-2.5 x 0.9-1.2 microm. The spores are oval with a characteristic surface layer, showing ornamentation-like projections, which are in close contact to the exospore. A short polar filament forming three to four coils traverses the polaroplast with two lamellar layers. The ultrastructure and other characteristic features of this microsporean parasite are distinct from those of the microsporean species described so far from oligochaetes.     

The morphology of Trianchoratus aecleithrium Price et Berry, 1966 (Dactylogyridae, Monogenea) from a new host Trichogaster trichopterus trichopterus (Pallas, 1777)

The monogenean parasite Trianchoratus aecleithrium Price et Berry, 1966 is comparatively described from the gills of Trichogaster trichopterus trichopterus from aquarium in Czechoslovakia. T. trichopterus trichopterus represents a new host record for this parasite.     

Gill parasites of Cephalopholis argus (Teleostei: Serranidae) from Moorea (French Polynesia): site selection and coexistence

The distribution and coexistence of gill ectoparasites of 121 specimens of Cephalopholis argus Bloch et Schneider, caught between October 1994 and October 1995, were investigated. Adults of the monogenean Benedenia sp. and copepod Hatschekia sp., the larval caligid copepod Caligus sp. (copepodite and chalimus stages), and praniza larvae of the isopod Gnathia sp. were found. All species were aggregated within the host population. Infracommunities were poor, with only 40.5% of fish infected by two parasite species. Only two individual fish harboured all the parasite species observed at the component community level. Prevalences were less than 50% and mean intensities were low (less than 6 parasites/host). No dominant parasite species were observed in the host population. The spatial distribution of each parasite species was studied on different partitions of the gill arches. Adult parasite stages that are mobile showed much overlap in their distribution, whereas temporarily attached larvae of Caligidae were more site specific. Copepodite and chalimus larvae showed niche restriction that is probably due to gregarious behaviour. Positive associations between caligid larvae reflected intraspecific interaction for site and/or resources. Each of the Caligus sp. larval stages prefers specific sites, as do the adults, which occur exclusively in the buccal cavity of the host. Infracommunities were too poor and too few to induce processes of interspecific competition.     

Soil-borne spores as a source of inoculum for wheat bunt (Tilletia caries)

Wheat bunt ( Tilletia caries ) has previously been controlled in the UK by routine seed treatment with organomercury. Recent examples are presented of unusual levels of infection due to soil-borne inoculum. The recent infections may be due to unusually dry weather, favouring the survival of spores in the soil, or to the presence of a more persistent strain of T. caries.     

Gyrodactylus bohemicus sp. n. (Monogenea: Gyrodactylidae) from Oncorhynchus mykiss (Walbaum) and Salvelinus fontinalis (Mitchill) (Clupeiformes: Salmonidae) in Czechoslovakia.

Gyrodactylus bohemicus sp. n (Gyrodactylidae: Monogenea) is described from the fins, skin and gill filaments of Oncorhynchus mykiss (Walbaum) and Salvelinus fontinalis (Mitchill) (Clupeiformes: Salmonidae) from a trout farm in Czechoslovakia. G. bohemicus sp. n. is most closely related to G. thymalli Zitnan, 1960 and G. magnus Konovalov, 1967 in the shape of the anchors, the ventral and the dorsal bar, but can be distinguished from both these species by the shape of the marginal hook proper.     

The microecology of dactylogyrids (Monogenea: Dactylogyridae) on the gills of wild spotted rose snapper Lutjanus guttatus (Lutjanidae) from Mazatlan Bay, Mexico

The spatial distribution and coexistence of monogenean dactylogyrids was assessed on the gills of 63 specimens of wild spotted rose snapper, Lutjanus guttatus (Steindachner), caught in the Mazatlan Bay, Sinaloa, Mexico. Five species are reported: Euryhaliotrema perezponcei Garcia-Vargas, Fajer-Avila et Lamothe-Argumedo, 2008, Euryhaliotrematoides sp., Haliotrematoides spinatus Kritsky et Mendoza-Franco, 2009, H. plectridium Kritsky et Mendoza-Franco, 2009, and H. guttati Garcia-Vargas, Fajer-Avila et Lamothe-Argumedo, 2008. All except E. perezponcei and H. guttati represent new geographical records for the Pacific coast. The most prevalent dactylogyrid species was E. perezponcei (100%), H. plectridium and H. spinatus had > 80% prevalence, and H. guttati and Euryhaliotrematoides sp. had the lowest prevalence. The mean abundance of H. plectridium and E. perezponcei was close to 60 parasites/fish, whereas Euryhaliotrematoides sp. and H. guttati had the lowest abundance. The dactylogyrid species exhibited a tendency for attachment to gill arch 2: 25% attachment occurring on gill arch 1, 30% on 2, 27% on 3 and 18% on 4, and showed a significant preference for the central sector of the gill (42%). Haliotrematoides plectridium had a preference for attachment to gill arches 2 and 3 and the central sector. Haliotrematoides spinatus tended to settle on the gill arches 2 and 3 and had a preference for the central sector. Euryhaliotrema perezponcei tended to settle on the gill arches 1 and 2 and the anterior gill sector. Euryhaliotrematoides sp. and H. guttati did not show a preference for any gill arch or sector. The intraspecific aggregation was stronger than the interspecific aggregation, indicating that all the dactylogyrid species on spotted rose snapper were aggregated, and there was no evidence of competition among the species.