Immunohistochemistry, histopathology and ultrastructure of Gasterosteus aculeatus tissues infected with Glugea anomala

Abstract

Immunohistochemical and histopathological studies were conducted on a population of 3-spined sticklebacks Gasterosteus aculeatus (L.) from Loch Airthrey (Stirling, Scotland) naturally infected with the microsporean Glugea anomala (Moniez 1887). Of the 55 host specimens that were examined, 16 (29.09%) were infected, the intensity of infection ranging from 1 to 4 xenomas per fish, which were principally located within the central portion of the body lateral flank musculature. All 32 G. anomala xenomas examined were mature, their diameter ranging from 936 to 2232 µm, and their walls of presented a laminar structure. Subcutaneously situated xenomas protruded from the fish body surface, whilst xenomas encountered within the intestine were seen to cause distortion. Light and electron microscopical observations confirmed a host cellular reaction around the xenoma, seen by the presence of eosinophile granule cells (EGCs), and some neutrophils. The occurrences of rodlet cells among the intestinal epithelial cells, and in close proximity to the xenoma wall, were observed in certain specimens. Outside the xenoma wall, macrophage aggregates (MAs) were commonly encountered. Within the xenoma wall, the presence of eosinophile granular cells immunoreactive to the anti-serotonin serum was also recorded. Further immunohistochemical tests revealed that a high number of nerve fibres running along the white lateral muscle fibres were immunoreactive to bombesin-, galanin-, and leu-enkephalin-antisera. Nerve fibres containing bombesin- and leu-enkephalin-like substances were also observed in the connective inflammatory tissue around the protozoan cyst, while neurons in the spinal ganglia were immunoreactive to met-enkephalin, and serotonin antisera. The control for the specificity of immunohistochemical reactions was performed using preabsorption tests of each antiserum with the corresponding antigen, and no immunoreactivity was noticed. The data presented are discussed in relation to the occurrence of G. anomala, which alters the pattern of nerve fibres present in the host. Specifically, the protozoan induces a response in the stickleback nervous system, the reaction of which is revealed through the application of immunohistochemical techniques.