Cytometric analysis of peripheral blood and of oxygen burst in neutrophilic granulocytes of healthy corps (Cyprinus carpio)

Flow cytometry is one of the most modern diagnostic techniques. It provides the potential for a rapid and multidimensional evaluation of properties of cells and their components. Present studies included the qualitative and q uantitative cytometric analysis of peripheral blood leukocytes and of oxyge n bursts in neutrophilic granulocytes of healthy carp in various stages of their individual development. Three populations of leukocyte line cells wer e distinguished, differing in their forward scatter (FSC) and side scatter (at 90 degrees, SSC) of the light beam. The most abundant pool of leukocyte s was present In the oldest carps (23 to 29 months old). The leukocytes wer e less numerous in the youngest carps (3 to 9 months of life) and were leas t numerous in carps of a moderate age (11 to 21 months old). Based on FSC/S SC characteristics, the distinguished leucocyte populations were found to c orrespond to lymphocytes/thrombocytes (low FSC, low SSC), granulocytes (hig h FSC, high SSC) and to monocytes (high FSC, low SSC). Cytometric analysis of the oxygen bursts in neutrophilic granulocytes of ca rp demonstrated increased values of the mean fluorescence channel in PMA st imulated nentrophilic granulocytes as compared to the values obtained for t he non-activated granulocytes or nentrophilic granulocytes which failed to become activated in response to the applied stimulator. Compared to the mea n fluorescence channel in cells not stimulated, by PMA, fluorescence intens ity was most pronounced in carps aging 11 to 21 months, less pronounced in the youngest carp (3 to 9 months old) and the least pronounced in the oldes t carps (23 to 29 months old). In the case of the analysis of the mean fluo rescence channel of a selected fraction of PMA-stimulated neutrophilic gran ulocytes as compared to the mean fluorescence channel of granulocytes which failed to respond by the stimulation reaction, the most pronounced fluores cence was noted to develop in the oldest carps, i.e. in carp aged 23 to 29 months. In the remaining carp the difference in the mean fluorescence chann el was significantly Simultaneously, the highest proportion of PMA-stimulat ed granulocytes was found to be present In 11 to 21-month-old fish, signifi cantly lower in the youngest (3 to 9-month-old) fish and the lowest in the oldest (23 to 29 months) fish.