Flow cytometry analyses of phagocytic and respiratory burst activities and cytochemical characterization of leucocytes isolated from wrasse (Labrus bergylta A.)

• Flow cytometry detection of phagocytic and respiratory burst activity of isolated leucocytes from wrasse (Labrus bergylta A.).
• Phenotypic characterization of leucocytes by cytochemical staining.
• Many eosinophils containing red granules were present among leucocytes isolated from head kidney, peripheral blood and spleen.

We have isolated leucocytes from peripheral blood (PBL), head kidney (HKL) and spleen (SL) of wrasse (Labrus bergylta A.) and studied the innate immune responses phagocytosis and respiratory burst using flow cytometry. Further, we have characterized the phenotypic properties of the leucocytes by cytochemical staining. We could differentiate between several subsets of leucocytes; lymphocytes, monocytes/macrophages, neutrophils, eosinophils, basophils and small leucocytes that might be precursor or immature cells. One striking observation was the eosinophils which were present among HKL, PBL and SL. The neutrophils had rounded, bean shaped or bi-lobed nuclei and resembled neutrophils in Atlantic cod (Gadus morhua L.) and lumpsucker (Cyclopterus lumpus L.), but were different from the polymorphonucleated neutrophils in Atlantic salmon (Salmo salar L.) and humans. Basophils were observed, but they were rare. Phagocytosis and respiratory burst activities were detected among different cell types. Highest phagocytic activity was observed among monocytes/macrophages and small leucocytes. Several different subtypes had ability to perform an oxygen-dependent degradation of microbes, measured as respiratory burst activity. Knowledge of the basic properties of wrasse's leucocytes and innate immunology can benefit further studies on its adaptive immune responses.

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