Journal: Animals : an Open Access Journal from MDPI

Article Title: The Effects of β-Hydroxy-β-Methylbutyrate (HMB) on Chemotaxis, Phagocytosis, and Oxidative Burst of Peripheral Blood Granulocytes and Monocytes in Goats

doi: 10.3390/ani9121031

Figure Lengend Snippet: Gating strategy for analysing flow cytometry data. Granulocytes and monocytes were gated based on forward and side scatter (FSC/SSC) parameters. Each cell subset was analysed for the relative number of phagocytising cells and cells stimulated for respiratory burst (fMLP, PMA, or E. coli bacteria). fMLP: N-formyl-methionyl-leucyl-phenylalanine, PMA: 4-phorbol-12-β-myristate-13-acetate.

Article Snippet: 100 μL of whole heparinised blood chilled to 0 °C and 20 μL of chilled Escherichia coli bacteria (Orpegen Pharma, Heidelberg, Germany) were added to each of the two 5 mL test tubes (blue, Beckman Coulter, Fullerton, CA, USA) (negative control and experimental sample) and were shaken for around 3 s at low speed.

Techniques: Flow Cytometry, Bacteria

Journal: Animals : an Open Access Journal from MDPI

Article Title: The Effects of β-Hydroxy-β-Methylbutyrate (HMB) on Chemotaxis, Phagocytosis, and Oxidative Burst of Peripheral Blood Granulocytes and Monocytes in Goats

doi: 10.3390/ani9121031

Figure Lengend Snippet: Percentage of granulocytes stimulated to undergo respiratory burst in 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 control and experimental group within the same sampling day at * p < 0.05; ** p < 0.01; **** p < 0.0001; A, B, C, D refer to statistically significant differences between day “0” and the consecutive sampling days within experimental group at A: p < 0.05; B: p < 0.01; C: p < 0.001; D: p < 0.0001.

Article Snippet: 100 μL of whole heparinised blood chilled to 0 °C and 20 μL of chilled Escherichia coli bacteria (Orpegen Pharma, Heidelberg, Germany) were added to each of the two 5 mL test tubes (blue, Beckman Coulter, Fullerton, CA, USA) (negative control and experimental sample) and were shaken for around 3 s at low speed.

Techniques: Control, Standard Deviation, Sampling

Journal: Animals : an Open Access Journal from MDPI

Article Title: The Effects of β-Hydroxy-β-Methylbutyrate (HMB) on Chemotaxis, Phagocytosis, and Oxidative Burst of Peripheral Blood Granulocytes and Monocytes in Goats

doi: 10.3390/ani9121031

Figure Lengend Snippet: Dot plot cytogram showing the percentage of granulocytes stimulated to undergo respiratory burst in control and experimental goats 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 stimulus), PMA (strong stimulus), or fMLP (weak stimulus) and incubated with dihydrorhodamine 123 in a water bath at a temperature of 37 °C. After incubation, cells were lysed and the DNA staining solution was added. The percentages of granulocytes stimulated to undergo respiratory burst (conversion of dihydrorhodamine 123 to rhodamine 123) were gated.

Article Snippet: 100 μL of whole heparinised blood chilled to 0 °C and 20 μL of chilled Escherichia coli bacteria (Orpegen Pharma, Heidelberg, Germany) were added to each of the two 5 mL test tubes (blue, Beckman Coulter, Fullerton, CA, USA) (negative control and experimental sample) and were shaken for around 3 s at low speed.

Techniques: Control, Negative Control, Bacteria, Incubation, Staining

Journal: Animals : an Open Access Journal from MDPI

Article Title: The Effects of β-Hydroxy-β-Methylbutyrate (HMB) on Chemotaxis, Phagocytosis, and Oxidative Burst of Peripheral Blood Granulocytes and Monocytes in Goats

doi: 10.3390/ani9121031

Figure Lengend Snippet: Mean fluorescence intensity (MFI) of granulocytes in 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 control and experimental group within the same sampling day at **** p < 0.0001; D refer to statistically significant differences between day “0” and the consecutive sampling days within experimental group at D: p < 0.0001.

Article Snippet: 100 μL of whole heparinised blood chilled to 0 °C and 20 μL of chilled Escherichia coli bacteria (Orpegen Pharma, Heidelberg, Germany) were added to each of the two 5 mL test tubes (blue, Beckman Coulter, Fullerton, CA, USA) (negative control and experimental sample) and were shaken for around 3 s at low speed.

Techniques: Fluorescence, Control, Standard Deviation, Sampling

Journal: Animals : an Open Access Journal from MDPI

Article Title: The Effects of β-Hydroxy-β-Methylbutyrate (HMB) on Chemotaxis, Phagocytosis, and Oxidative Burst of Peripheral Blood Granulocytes and Monocytes in Goats

doi: 10.3390/ani9121031

Figure Lengend Snippet: Percentage of monocytes stimulated to undergo respiratory burst in 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 control and experimental group within the same sampling day at * p < 0.05; A and B refer to statistically significant differences between day “0” and the consecutive sampling days within experimental group at A: p < 0.05; B: p < 0.01.

Article Snippet: 100 μL of whole heparinised blood chilled to 0 °C and 20 μL of chilled Escherichia coli bacteria (Orpegen Pharma, Heidelberg, Germany) were added to each of the two 5 mL test tubes (blue, Beckman Coulter, Fullerton, CA, USA) (negative control and experimental sample) and were shaken for around 3 s at low speed.

Techniques: Control, Standard Deviation, Sampling

Journal: Animals : an Open Access Journal from MDPI

Article Title: The Effects of β-Hydroxy-β-Methylbutyrate (HMB) on Chemotaxis, Phagocytosis, and Oxidative Burst of Peripheral Blood Granulocytes and Monocytes in Goats

doi: 10.3390/ani9121031

Figure Lengend Snippet: Mean fluorescence intensity (MFI) of monocytes in 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 control and experimental group within the same sampling day at **** p < 0.0001; D refer to statistically significant differences between day “0” and the consecutive sampling days within experimental group at D: p < 0.0001.

Article Snippet: 100 μL of whole heparinised blood chilled to 0 °C and 20 μL of chilled Escherichia coli bacteria (Orpegen Pharma, Heidelberg, Germany) were added to each of the two 5 mL test tubes (blue, Beckman Coulter, Fullerton, CA, USA) (negative control and experimental sample) and were shaken for around 3 s at low speed.

Techniques: Fluorescence, Control, Standard Deviation, Sampling

Journal: Animals : an Open Access Journal from MDPI

Article Title: The Effects of β-Hydroxy-β-Methylbutyrate (HMB) on Chemotaxis, Phagocytosis, and Oxidative Burst of Peripheral Blood Granulocytes and Monocytes in Goats

doi: 10.3390/ani9121031

Figure Lengend Snippet: Dot plot cytogram showing the percentage of phagocytic granulocytes in control and experimental goats on experimental day 30. Whole heparinised blood from control and experimental animals was incubated for 10 minutes with an FITC-labelled E. coli in an ice bath at a temperature of 0 °C (negative control) or in a water bath at a temperature of 37 °C (control and HMB). The percentages of granulocytes with ingested E. coli (FITC) bacteria were gated.

Article Snippet: 100 μL of whole heparinised blood chilled to 0 °C and 20 μL of chilled Escherichia coli bacteria (Orpegen Pharma, Heidelberg, Germany) were added to each of the two 5 mL test tubes (blue, Beckman Coulter, Fullerton, CA, USA) (negative control and experimental sample) and were shaken for around 3 s at low speed.

Techniques: Control, Incubation, Negative Control, Bacteria

Journal: Animals : an Open Access Journal from MDPI

Article Title: The Effects of β-Hydroxy-β-Methylbutyrate (HMB) on Chemotaxis, Phagocytosis, and Oxidative Burst of Peripheral Blood Granulocytes and Monocytes in Goats

doi: 10.3390/ani9121031

Figure Lengend Snippet: Dot plot cytogram showing the percentage of phagocytic monocytes in control and experimental goats on experimental day 30. Whole heparinised blood from control and experimental animals was incubated for 10 minutes with FITC-labelled E. coli in an ice bath at a temperature of 0 °C (negative control) or in a water bath at a temperature of 37 °C (control and HMB). The percentages of monocytes with ingested E. coli (FITC) bacteria were gated.

Article Snippet: 100 μL of whole heparinised blood chilled to 0 °C and 20 μL of chilled Escherichia coli bacteria (Orpegen Pharma, Heidelberg, Germany) were added to each of the two 5 mL test tubes (blue, Beckman Coulter, Fullerton, CA, USA) (negative control and experimental sample) and were shaken for around 3 s at low speed.

Techniques: Control, Incubation, Negative Control, Bacteria