[A morphofunctional analysis of the hemocytes in the cricket Gryllus bimaculatus (Orthoptera: Gryllidae) normally and in acute microsporidiosis due to Nosema grylli].

Microsporidians (M) are supposed to be ancient eukaryotic parasites with a broad range of animal hosts, being especially abundant in Arthropoda. They are supposed to pass a long way of adaptation to parasitism, that usually means inhibiting or avoiding host immune reactions alongside with the reduction of pathogenicity. However M, unlike other eukaryotic obligate parasites, preserved a high pathogenicity, comparable with one of viruses, and thus they could be expected to possess a unique mode of interactions with their hosts. The goal of the present work is to assess how M influence the cellular immune response of an insect host. Experiments were performed on the host-parasite system Gryllus bimaculatus (Orthoptera, Gryllidae)--Nosema grylli (Microsporidia, Nosematidae); coccidia Adelina grylli--infected crickets were used to compare the host cellular response against two pathogens. Haemocytes (H) were observed using phase contrast and electron microscope. H smears were stained for a phenoloxidase (PO), esterase activities and "respiratory burst" reaction. Five H type can be distinguished in the cricket haemolymph. (1) Prohaemocytes, relatively small (13-30 microns) cells with large nuclei, are observed both in control and infected insects. (2) Plasmatocytes, round (30-35 microns in diameter) or fusiod (40-63 x 13-38 microns) cells, can hardly be distinguished from (1) ultrastructurally; during the coccidian infection of the cricket fat body these H infiltrate the infected organ and turn into amebocytes with laciniate nuclei, they usually contain electron dense granules, that release during the formation of a capsule around the coccidian oocytes. (3) Granulocytes (Gr, 20-33 microns in diameter) are cells with the extremely refractive cytoplasm when observed in phase contrast microscope, they contain vacuoles with typical crystal needle-like inclusions. The transitional forms between the mentioned above three cell types can be observed. The next two H types also observed on H monolayers are supposed to be the specialized forms of Gr: (4) coagulocytes, cells with the fragile cytoplasm that are easily disintegrated after a contact with a pathogen; they have been described in Orthoptera for the first time now; (5) spherulocytes, giant cells filled with electron lucid granules and small, often eccentrically located nucleus. Both H types were observed only after infection with A. gryllus in the vicinity of encapsulated oocysts. Infection with M does not cause such intensive concentration of haemocytes near the infected organ, or so abundant nodule formation, until the acute stage of the disease when M spores are liberated from the destroyed cells and contact the insect haemolymph. Thereafter, the number of granulocytes significantly increases. In the presence of M spores, haemocytes produce long cytoplasm protrusions and form clapms. Some spores adhere to the haemocyte surface and are phagocytized. Giant round cells loaded with spores, can be observed in the host lymph. They are surrounded by a sheath composed of flattened cells and resemble xenomas, described for fish microsporidiosis. A. grylli caused the increase in the quote of PO-positively stained cells up to 80% from 40-50% in control, that well corresponds to the host immune reactions activation and melanization of infected tissue, while microsporidiosis significantly reduced quote of PO+ cells. Carboxyl esterase activity expressed as quote of positively stained cells was 40-60% in naive and coccidia-parasitized samples, M decrease this number to 10-20%. "Respiratory burst" reaction, detected by reducing of NBT, did not alter significantly in microsporidia-infected insects. From the presented data in can be concluded: 1) M do not suppress such cellular reactions as a clamp formation and phagocytosis of spores, liberated from the infected tissue; 2) at the same time they suppress activities of enzymes involved in immune response.