Fig. 4
a The percentage of granulocytes stimulated to undergo respiratory burst in calf groups after stimulation with fMLP, PMA and E. coli, as determined in the Bursttest® kit. Key: I – control group; II – experimental group; SD - standard deviation. Numerical results were presented as the arithmetic mean ± SD. The significance level was set at 0.05. Asterisks refer to statistically significant differences between the control group and the experimental group on the same sampling day at * p < 0.05; ** p < 0.01; A, B refer to statistically significant differences between day “0” and the consecutive sampling days in the experimental group at A – p < 0.05; B – p < 0.01. b. Dot plot cytogram showing the percentage of granulocytes stimulated to undergo respiratory burst in control and experimental calves on experimental day 30. Whole heparinised blood from control and experimental animals (control and HMB) was divided into four test tubes. The samples were combined with the washing solution (negative control), E. coli bacteria (opsonising activator), PMA (strong activator) or fMLP (weak activator) and incubated with dihydrorhodamine 123 in a water bath at a temperature of 37 °C. After incubation, cells were lysed and DNA staining solution was added. The percentages of granulocytes stimulated to undergo respiratory burst (conversion of dihydrorhodamine 123 to rhodamine 123) were gated. c. Mean fluorescence intensity (MFI) of granulocytes in calf groups after stimulation with fMLP, PMA and E. coli, as determined in the Bursttest® kit. Key: I – control group; II – experimental group; SD - standard deviation. Numerical results were presented as the arithmetic mean ± SD. The significance level was set at 0.05. Asterisks refer to statistically significant differences between the control group and the experimental group on the same sampling day at * p < 0.05; ** p < 0.01; *** p < 0.001; **** p < 0.0001; B, C, D refer to statistically significant differences between day “0” and the consecutive sampling days in the experimental group at B – p < 0.01; C – p < 0.001; D – p < 0.0001