1. OmniGen-AF® supplementation improves leukocyte responses and hematology in multiparous peripartum cows
C.R. Nightingale1, M.D. Sellers1, A.R. Pepper-Yowell1, J.D. Chapman2 and M.A. Ballou1
1Texas Tech University, Lubbock
2Phibro Animal Health Corporation, Quincy, IL
1/12/2019

2. Objective
To evaluate changes in indicators of immunological stress both pre- and postpartum in healthy multiparous dairy cows supplemented with OmniGen-AF from dry-off through 28 DIM
1/12/2019

3. Experimental Design
Location: Texas Tech U. & a commercial Holstein dairy
Design:
Control (CT) or basal diets (no OmniGen-AF) vs. basal diets + OmniGen- AF (OG, 56g/h/d)
Diets:
Diets fed as TMR’s (far-off, close-up negative DCAD, lactation)
OmniGen-AF pre-blended with wheat middlings to provide 56g/h/d in a lb./d feeding rate
OmniGen-AF blend was added on-farm to respective TMR’s after control cows were fed
Diets fed beginning at dry-off through 28 DIM
1/12/2019

4. Experimental Design Metrics: Neutrophil response Inflammatory Markers
L-selectin (CD62L) expression, oxidative burst capacity
Neutrophil to Lymphocyte ratio
Inflammatory Markers
Haptoglobin
Tumor Necrosis Factor-Alpha
Blood metabolites
NEFA, BHBA, blood urea nitrogen (BUN)
Milk & milk components: tested at 30 d intervals
Milk yield, 2x milking
Milk fat & milk protein (% and lb.)
Somatic cell count
1/12/2019

5. Experimental Design Metrics: Neutrophil response Inflammatory Markers
L-selectin (CD62L) expression, oxidative burst capacity
Neutrophil to Lymphocyte ratio
Inflammatory Markers
Haptoglobin
Tumor Necrosis Factor-Alpha
Blood metabolites
NEFA, BHBA, blood urea nitrogen (BUN)
Milk & milk components: tested at 30 day intervals
Milk yield, 2x milking
Milk fat & milk protein (% and kg.)
Somatic cell count
1/12/2019

6. Experimental Design Cow selection: Data collection:
Cows enrolled at dry-off using Dairycomp305 records based on parity (avg. 2.55), expected calving date (-60 ± 3 d) and previous production and health history
Only clinically healthy cows that successfully completed the entire trial period were used in the final statistical analyses (CT=24, OG=23)
Data collection:
Blood samples collected: at dry-off, mid-dry, calving and 14 and 28 DIM
Statistics: Linear mixed model with repeat measures using baseline (-60 d) measures as a covariate
1/12/2019

7. Figure 1. Neutrophil surface expression of L-selectin**
Reference:
J. Dairy Sci., vol. 96, E suppl 1 (abstract & poster) - OmniGen-AF supplementation improves leukocyte responses and hematology of multiparous peripartum cows.
L-selectin measures as surface bound protein. Used Ab to measure l-selectin (GMFI = geometric fluoresence intensity).
Graph illustrates the CD62L (L-selectin) profiles from OG and control cows from -30 to +28 days post-partum. OG cows had significantly greater levels pre-partum and at calving. These data mirror those reported by Burton et al. for the control cows and reflect the ‘rescue’ effect observed by Mullarky et al.
FYI: GMFI = geometric fluorescense intensity
**P<0.001
1/12/2019

8. Figure 2. Neutrophil oxidative burst capacity*
Reference:
J. Dairy Sci., vol. 96, E suppl 1 (abstract & poster) - OmniGen-AF supplementation improves leukocyte responses and hematology of multiparous peripartum cows.
Graph illustrates oxidative bursts of neutrophils. Oxidative bursts are the release of reactive oxygen species (ROS), chemical compounds used to kill pathogens. Although effective in destroying harmful pathogen, excessive production of ROS can damage healthy tissue (ie: mammary gland). These data suggest that OG is reducing or attenuating this release of ROS thereby potentially avoiding tissue damage while still effectively able to ‘kill’ pathogens.
To test for oxidative burst capacity, neutrophils were harvested from OG and control cows and incubated with E. coli until 50% of neutrophils produced an oxidative burst.
1/12/2019

9. Figure 3. Plasma haptoglobin concentrations**
Reference:
J. Dairy Sci., vol. 96, E suppl 1 (abstract & poster) - OmniGen-AF supplementation improves leukocyte responses and hematology of multiparous peripartum cows.
Haptoglobin (Hp) is an acute phase protein produced in response to any inflammation process (infection, stress, burns, injury, etc.) or an immunological challenge (calving). Hp is a measure of systemic inflammation. In this study Nightingale et al., observed Hp response to be less in the OG fed cows verses the controls at d 14 post-partum, suggesting that OG may be attenuating the inflammation response.
1/12/2019

10. Figure 4. Tumor necrosis factor-alpha
#P<0.10, **P<0.048 # **
Reference:
J. Dairy Sci., vol. 96, E suppl 1 (abstract & poster) - OmniGen-AF supplementation improves leukocyte responses and hematology of multiparous peripartum cows.
Tumor Necrosis Factor alpha, involved in systemic inflammation, is a member of a group of cytokines that stimulate the acute phase reaction. Produced chiefly by macrophages and other immune cells, CD4+lymphocytes, NK cells and neurons. TNF-alpha primary role is regulation of immune cells. TNF an endogenous pyrogen (fever inducer), apoptoic cell death, cachexia (loss of body mass) and inflammation.
1/12/2019

11. Milk and Milk Components
Control
OmniGen-AF
Milk, lb.
First Test
104.3
103.0
Avg. All Tests
98.3
98.8
Fat, %
3.82
3.74
3.37
3.38
Fat, lb.
3.98
3.30
3.32
Protein, %
2.95
2.94
2.92
Protein, lb.
3.06
2.88
No differences were detected between the control or OmniGen-AF fed cows for milk or milk components. The ‘average all tests’ for milk and components is calculated from 5 consecutive test-days (30 through 150 DIM).
1/12/2019

12. Milk and Milk Components
Control
OmniGen-AF
Milk, kg.
First Test
47.41
46.82
Avg. All Tests
44.68
44.90
Fat, %
3.82
3.74
3.37
3.38
Fat, kg.
1.81
1.50
1.51
Protein, %
2.95
2.94
2.92
Protein, kg.
1.39
1.31
No differences were detected between the control or OmniGen-AF fed cows for milk or milk components. The ‘average all tests’ for milk and components is calculated from 5 consecutive test-days (30 through 150 DIM).
1/12/2019

13. Milk Yield, lb. www.theOmiGenDifference.com Reference:
J. Dairy Sci., vol. 96, E suppl 1 (abstract & poster) - OmniGen-AF supplementation improves leukocyte responses and hematology of multiparous peripartum cows.
1/12/2019

14. Milk Yield, kg. www.theOmiGenDifference.com Reference:
J. Dairy Sci., vol. 96, E suppl 1 (abstract & poster) - OmniGen-AF supplementation improves leukocyte responses and hematology of multiparous peripartum cows.
1/12/2019

15. Somatic Cell Count * * Test-day 120 , SCC’s differed P = 0.024
Reference:
J. Dairy Sci., vol. 96, E suppl 1 (abstract & poster) - OmniGen-AF supplementation improves leukocyte responses and hematology of multiparous peripartum cows.
No differences in SCC were detected across all test-days (CT=131,000 cell/ml vs. OG=54,000 cell/ml, P=0.065), however SCC’s at test-day 120 were different (P=0.024).
* Test-day 120 , SCC’s differed P = 0.024
1/12/2019

16. Summary
Cows fed OmniGen-AF (OG) from dry off through 28 day in milk were observed to have a more functional innate immune response during the transition period as measured by:
A greater surface expression of neutrophil L-selectin (P < 0.001)
A lower neutrophil to lymphocyte ratio (P<0.001)
An elevated plasma concentrations of tumor necrosis factor-alpha (P < 0.10)
Haptoglobin concentrations were lower (P < 0.002) at 14 DIM in OG cows, suggesting an attenuation of the inflammation response associated with parturition
Milk or milk components were not different. Overall SCC’s tended (P < 0.065) to be lower for OG cows and were different (P < 0.024) at 120 DIM
1/12/2019