Josep M. Amigó

Longevity and effects of temperature on the viability and polar-tube extrusion of spores of Glugea stephani, a microsporidian parasite of commercial flatfish.

Abstract Food contamination with respect to microsporidiosis in humans (associated with acquired immunodeficiency syndrome) and in marine fish farming deserves particular attention. For the first time, a study on the longevity and resistance to both heat and freezing of the spores of a vertebrate microsporidian, Glugea stephani, parasitizing a commercial flatfish was carried out. As judged on the basis of determinations of the extrusion rate, the resistance of the spores to temperature stress was remarkable. The extrusion rate, which can be directly related to infectivity, was always lower than the viability (membrane integrity). It should be pointed out that neither heat (60° C for 30 min) nor freezing (−19° C for 24 h) caused a complete reduction in the extrusion rate or viability. Consequently, the ingestion of poorly cooked or raw fish (even if previously frozen) represents a danger for aquaculture and, probably, for immunodepressed patients.

Tetramicra brevifilum (Matthews & Matthews, 1980) (Microsporida: Tetramicriidae) in a new fish host, Lophius budegassa (Spinola, 1807) in Spain

Tetramicra brevifilum, a microsporidian parasite of Scophthalmus maximus, was found in Lophius budegassa for the first time. This parasite was detected in 5 of 199 hosts captured in the coastal waters of Barcelona (Northwest Mediterranean), which enlarges the geographic distribution of this microsporidian. Affected fish did not show any external sign of disease, and cysts of T. brevifilum were found associated with the body musculature but were easily differentiated from those of Spraguea lophii, another microsporidian present in this host. A case of simultaneous infection by both T. brevifilum and S. lophii was found.

Comparative Study of Microsporidian Spores by Flow Cytometric Analysis

ABSTRACT. Spore suspensions of microsporidian parasites of fish (Microsporidium ovoideum, Glugea stephani, Glugea atherinae and Spraguea lophii) have been analyzed by flow cytometry. Spore nuclei were dyed either by propidium iodide or bis‐benzimide (Hoechst 33342). By observation of forward light scatter and fluorescence the four species could be distinguished and the mono‐ and diplokaryotic populations of S. lophii identified. Staining of DNA by bis‐benzimide was better and easier than propidium iodide. Forward light scatter and fluorescence values were characteristic of each species and remained unchanged throughout the year, so flow cytometry can be used for distinction of spores of some microsporidian parasites once their flow cytometric parameters are known. However, special care has to be taken in tool calibration and material preparation for analysis because of the high precision of the technique.

Ultrastructure and development of Microsporidium ovoideum (Thélohan, 1895) Sprague, 1977, a microspohdian parasite of the red band fish (Cepola macrophthalma L): Redescription of the organism and reassignment to the genus Microgemma, ralphs & matthews 1986

Summary A study of the ultrastructure and life cycle of Microsporidium ovoideum , a hepatic parasite of Cepola macrophthalma , has been carried out. This parasite caused xenomas up to 1–2 mm, which consisted of a hypertrophic cell, filled with different stages of the parasite and rests of the host cell organelles, which had a pychnotic hypertrophic nucleus. Merogonic stages consisted of round multinucleate plasmodia that divided by plasmotomy or exogenous budding, and were not enclosed by any membrane. Sporogonic plasmodia divided by exogenous budding to produce sporoblast mother cells which after a binary fission become sporoblasts. Sporogony occurs in direct contact with host cytoplasm. The spores, which are ovoid and uninucleate, show a posterior vacuole in the posterior third of the spore and an isofilar polar filament with 6–8 coils. The characteristic features permit a reassignation of this microsporidium into the genus Microgemma and give a new combination: Microgemma ovoidea .

The pathologic cycle of the infection of the microsporidian Microgemma ovoidea (Thèl., 1895) Amigó et al. 1996 in the liver of the Red Band fish (Cepola macrophthalma L.).

Abstract Pathologic study of the lesions caused by Microgemma ovoidea has shown that after the formation of the xenoma (stage 1), the parasitized cell is infiltrated by host macrophages (stage 2) and quickly encysted by the activity of fibroblasts that form a xenoma wall composed of collagenous fibers (stage 3). The phagocytic activity of the macrophages leads to the formation of a granuloma (stage 4) in which the cyst contents comprise macrophages filled with phagocytosed spores. This phagocytic activity is limited by the fact that some parts of the microsporidian spores, such as the spore walls, cannot be lysed by macrophages, which leads to the formation of fused giant cells containing nondigestible spore remnants. The final step in the process is healing (stage 5), in which some cells may start proliferating to regenerate the damaged area. Nevertheless, the host occasionally fails to control M. ovoidea infections. This failure can take two forms: bursting of the granuloma, or the appearance of secondary infections in granulomas, probably through parasitism of macrophages.