Dr. Milo Wiltbank presented this for a DAIReXNET webinar on Wednesday, March 2, 2016. The full presentation recording can be found at http://bit.ly/1wb83YV.

1. Milo C. Wiltbank1, Paulo D. Carvalho1, Alex H. Souza1,
Paul M. Fricke1, Mateus Z. Toledo1, Roberto Sartori2,
Jose E. Santos3, Guillerme Pontes2, and R. D. Shaver1
1Department of Dairy Science, University of Wisconsin-
Madison
2Department of Animal Sciences, University of São
Paulo, Brazil
3Department of Animal Science, University of Florida,
USA
Potential benefits of nutrition on reproductive
performance of high-efficiency dairy cows.

2. What is the role of nutrition in improving
reproductive efficiency in today’s dairy herds?
Improving
Reproduction
Genetics
Nutrition
Management
Physiology
Genomics allow selection of cows that are
high for DPR and Cow conception rate.
Reproductive
management
programs are
available that drive
service rate and
also improve
fertility.
Cow comfort,
compliance, and
AI skills are all
essential for
reproductive
management.

3. Critical Periods for Nutritional
Effects on Reproduction
Dry Period
(last 3 wks)
Early Post-
partum (3 wks)
Pre-AI
(1 wk)
Pregnancy
What is the role of nutrition in improving
reproductive efficiency in today’s dairy herds?

4. Effect of Injectable Vitamin E on Incidence of Retained Fetal
Membranes and Reproductive Performance of Dairy Cows
G.C.S. Pontes*, P.L.J. Monteiro Jr*, A.B. Prata*, M.M. Guardieiro*, D.A.M.
Pinto*, G. Fernandes*, M.C. Wiltbank*,§, J.E.P. Santos¶, and R. Sartori*
*Department of Animal Sciences, University of São Paulo, ESALQ, Piracicaba,
Brazil 13418-900
§Department of Dairy Sciences, University of Wisconsin, Madison, 53706
¶Department of Animal Sciences, University of Florida, Gainesville, FL 32611-
0910
Vitamin E before calving

5. Three dairy farms in Brazil (n = 890 cows)
Cows received 280 (Farm 1), 390 (Farm 2), and 480
(Farm 3) IU of vitamin E/d as part of the grain mix
during dry period.
This was inadequate (< 50% of recommendations from
NRC) and 53% had inadequate serum Vitamin E.
Cows given nothing (Controls) or 1,000 IU of vitamin E
at 258±3, 265±3, and 272±3 d of gestation (~3, 2,
and 1 week before expected calving)
Methods

6. 2.00
2.25
2.50
2.75
3.00
3.25
3.50
3.75
4.00
α-tocopherol,μg/mL
Day relative to calving
0
0
0
0
0
0
0
0
0
0
ProbabilityofRFM
A
1000 IU Vitamin E
Vitamin E before calving
Vitamin E decreases as calving
approaches due to accumulation
in colostrum

7. 0.00
0.05
0.10
0.15
0.20
0.25
0.30
0.35
0.40
0.45
ProbabilityofRFM
Serum α-tocopherol, μg/mL
B
Vitamin E before calving
Cows with lower Vitamin E (a-tocopherol)
had higher probability of retained fetal
membranes (RFM = retained placenta).

8. Item Control Vitamin E Risk P
Cows, n
441 449 ---- ----
Retained placenta,%
20.1 13.5 1.68 0.007
Stillbirth, %
14.9 6.8 2.42 0.001
Death by 200 DIM ,%
5.4 2.4 2.28 0.07
Milk yield,kg (± SEM)
60 DIM 1,398 ± 28 1,400 ± 28 ---- 0.96
100 DIM 2,425 ± 47 2,388 ± 46 ---- 0.46
305 DIM 6,478 ± 109 6,413 ± 108 ---- 0.59
Vitamin E before calving

9. Item Control Vitamin E Risk P
Cows3 362 344 ---- ----
Days at 1st AI (± SE) 64.3 ± 1.2 62.1 ± 1.3 ---- 0.12
First postpartum AI
Pregnant d 31, % 38.1 42.1 0.85 0.30
Pregnant d 62, % 30.1 36.7 0.76 0.08
Pregnancy loss, % 20.5 12.5 2.30 0.01
All AIs to 200 DIM
Pregnant d 31, % 34.5 38.4 0.59 0.02
Vitamin E before calving

10. JDS 96:5879; 2013
Retrospective evaluation of 7 previous
studies (n = 408 cows total) that were fed
controlled-energy or high energy diets during
the far-off or close-up dry period.

11. JDS 96:5879; 2013
Controlled
Energy Diet
High Energy
Diet
P Value
Days to first
AI
86 Days 95 Days P = 0.72
Days from
Calving to
Pregnancy
157 Days 167 Days P = 0.04
BCS Loss
week 1-6
after calving
-0.30 -0.43 P = 0.04
NEL for first
4 weeks of
lactation
22.8 Mcal/d 22.4 Mcal/d N.S.

12. Critical Periods for Nutritional
Effects on Reproduction
Dry Period
(last 3 wks)
Early Post-
partum (3 wks)
Pre-AI
(1 wk)
Pregnancy
What is the role of nutrition in improving
reproductive efficiency in today’s dairy herds?
- Supplemental vitamin E before calving can reduce:
Retained Fetal Membranes, Stillbirth, Pregnancy Loss
- Dry Period Nutrition can alter calving and early post-
partum physiology AND subsequent fertility.

13. Primordial
Germ Cell
Primordial
Follicle
Activation
Preantral to
Antral Follicle
Antral Follicle
Development
Ovulation
Fertilization
Morula
Blastocyst
Potential Causes of
Early Embryo Degeneration
Alterations in oocyte due
to nutritional effects near
calving?
Poorer
Quality
Embryos?
~60 days

14. Relationships between fertility and postpartum
changes in body condition and body weight in
lactating dairy cows.
Journal of Dairy Science 97:1-18.
2014
Carvalho, PD, Souza, AH, Amundson, MC, Hackbart,
KS, Fuenzalida, MJ, Herlihy, MM, Ayres, H, Dresch, AR,
Vieira, LM, Guenther, JG, Grummer, RR, Fricke, PM,
Shaver, RD, Wiltbank, MC.

15. Double-Ovsynch
= Treats Anovular Cows
Sun Mon Tue Wed Thu Fri Sat
GnRH
PGF
GnRH
GnRH
PGF GnRH TAI

16. BCS
2.25 2.50 2.75 3.00 3.25 3.50
Probabilityofconception
20
25
30
35
40
45
50
< >
Ovsynch (n = 1,081)
Souza et al., 2008
Body Condition Score at
Time of AI
Double-Ovsynch (n = 1,103)
Carvalho et al., 2014
~40%
~50%
P = 0.03
P = 0.03

17. Experiment 2
• 1,887 lactating dairy cows from 2 farms in WI
Synchronized with Double-Ovsynch for 1st service
(Carvahlo et al., 2014. Relationships between fertility and
postpartum changes in body condition and body weight in
lactating dairy cows. Journal of Dairy Science 97:1-18.)
• BCS evaluated at calving and 21 d later using a 5 point scale
with 0.25 increments (Edmonson et al., 1989)
- BCS change = BCS@calving vs. BCS@21DIM
divided in 3 groups:
- Lost BCS (41.8%; 789/1887)
- Maintained BCS (35.8%; 675/1887)
- Gained BCS (22.4%; 423/1887)
17

18. Pregnancies/AI
0
20
40
60
80
100
40 d 70 d
Lost Maintained Gained
BCS change: P < 0.001
Parity: P < 0.001
789 789675 675
25%c
23%c
38%b
36%b
Pregnancies/AI(%)
84%a
78%a
423 423
BCS change: P < 0.001
Parity: P < 0.001

19. Expt 3 - % Body Weight change
%BodyWeightChange
fromfirstweek
-10
-8
-6
-4
-2
0
2
4
6
First Quartile
Second Quartile
Third Quartile
Fourth Quartile
A
Group P < 0.001
Week P < 0.001
Group x Week P < 0.001
Week Postpartum
0 1 2 3 4 5 6 7 8 9 10
Superovulation &
Embryo Evaluation
n = 72 donors
560 embryos
-8% of Body Weight
-4% of BW
~0% change
+4% change

20. Embryo Characteristics
Quartile
P-
value
Fourth Q
Lost +
Third Q
Lost
Second Q
Maintain
First Q
Gain
CL (#) 18.4±2.6 18.4±1.7 19.0±1.7 16.0±2.0 0.67
Fertilized
(%)
76.9±7.1 77.0±6.6 77.6±7.6 78.4±7.1 0.99
Quality 1 &
2 (%)
38.0±8.7 61.3±8.2 60.6±9.4 63.4±8.6 0.14
Degenerate
(%)
35.2±8.5a 12.6±4.6b 14.5±6.3b 9.6±3.7b 0.02
Qual 1 & 2
of Fert (%)
48.4±9.5a 78.3±6.6b 72.6±9.5b 77.7±7.4b 0.05
Degenerate
of Fert (%)
46.9±9.6a 17.4±6.4b 24.8±9.3ab 16.2±7.0b 0.04
Summary: Cows that lose body weight during the
first 21 d after calving had much lower embryo
quality compared to cows that gain body weight.

21. Expt 3 - % Body Weight change%BodyWeightChange
fromfirstweek
-10
-8
-6
-4
-2
0
2
4
6
First Quartile
Second Quartile
Third Quartile
Fourth Quartile
A
Group P < 0.001
Week P < 0.001
Group x Week P < 0.001
Week Postpartum
0 1 2 3 4 5 6 7 8 9 10
Quality 1 & 2 of Fert
48%
78%
73%
78%
Limitations?
Did not manipulate the cows into the body weight change
groups but we let the cows divide themselves into groups.
Is this really BCS loss or early post-partum disease effects?

22. Critical Periods for Nutritional
Effects on Reproduction
Dry Period
(last 3 wks)
Early Post-
partum (3 wks)
Pre-AI
(1 wk)
Pregnancy
What is the role of nutrition in improving
reproductive efficiency in today’s dairy herds?
- Lower BCS near time of AI:
Reduces fertility (P/AI) but can be reduced with
Double-Ovsynch protocol
- Greater post-partum BCS loss:
Reduces fertility (P/AI) due to effects on early embryo

23. Primordial
Germ Cell
Primordial
Follicle
Activation
Preantral to
Antral Follicle
Antral Follicle
Development
Ovulation
Fertilization
Morula
Blastocyst
Causes of
Early Embryo Degeneration
Duration of Follicle Dominance
Progesterone environment
during follicle dominance
Nutritional Effects?
Why Embryos
Degenerate?

24. Objective: Evaluate association of individual
components of the diet on fertility traits of dairy
cattle.
Data: Complete diets were obtained and Dairy
Comp 305 backups for fertility and other traits
were evaluated in 50 dairy farms.

25. Diets in 50 herds vs Reproduction (DC305) records
Dietary Factor % Pregnant
at 150 DIM
P/AI at First
Service
P/AI at all
services
NDF = Non-
Detergent Fiber
r = 0.111
P = 0.48
r = 0.345
P = 0.025
r = 0.249
P = 0.11
NFC = Non-Fiber
Carbohydrate
r = -0.331
P = 0.03
r = -0.512
P = 0.0005
r = -0.476
P = 0.0015
Starch r = -0.156
P = 0.39
r = -0.348
P = 0.047
r = -0.194
P = 0.280
Fat r = 0.235
P = 0.135
r = 0.346
P = 0.02
r = 0.244
P = 0.12
Lysine, %
Metabolizable Protein
r = 173
P = 0.27
r = -0.003
P = 0.99
r = -0.079
P = 0.62
Methionine, %
Metabolizable Protein
r = 0.2814
P = 0.07
r = 0.104
P = 0.51
r = 0.0056
P = 0.972

26. 20.0
25.0
30.0
35.0
40.0
45.0
50.0
30.0 35.0 40.0 45.0 50.0
P/AIto1stpostpartumAI
Non Fiber Carbohydrate - NFC, %
Diets for 50 herds vs DC305 records
r = -0.51

27. Diets in 50 herds vs Reproduction (DC305) records
Dietary Factor % Pregnant
at 150 DIM
P/AI at First
Service
P/AI at all
services
NDF = Non-
Detergent Fiber
r = 0.111
P = 0.48
r = 0.345
P = 0.025
r = 0.249
P = 0.11
NFC = Non-Fiber
Carbohydrate
r = -0.331
P = 0.03
r = -0.512
P = 0.0005
r = -0.476
P = 0.0015
Starch r = -0.156
P = 0.39
r = -0.348
P = 0.047
r = -0.194
P = 0.280
Fat r = 0.235
P = 0.135
r = 0.346
P = 0.02
r = 0.244
P = 0.12
Lysine, %
Metabolizable Protein
r = 173
P = 0.27
r = -0.003
P = 0.99
r = -0.079
P = 0.62
Methionine, %
Metabolizable Protein
r = 0.2814
P = 0.07
r = 0.104
P = 0.51
r = 0.0056
P = 0.972

28. Critical Periods for Nutritional
Effects on Reproduction
Dry Period
(last 3 wks)
Early Post-
partum (3 wks)
Pre-AI
(1 wk)
Pregnancy
What is the role of nutrition in improving
reproductive efficiency in today’s dairy herds?
- Too high of carbohydrates in diet:
Reduces P/AI possibly due to elevated insulin
- Fat, particularly PUFA, can improve P/AI
- Methionine in diet can improve reproductive efficiency

29. Stops abnormal
Gene expression
Methionine
Nutritional Epigenomics

30. Material & Methods
 Holstein cows (n = 309)
 Milked twice daily (milk yield = 40.9 kg/d)
 Cows were blocked by parity and randomly
assigned to two treatments:
 Rumen-Protected methionine (RPM) - 2521 g
MP with HIS 2.72, LYS 6.93 & MET 2.34
(%MP)
 Control (CON) - 2521 g MP with HIS 2.72, LYS
6.93 & MET 1.87 % (% MP)

31. Feeding Treatments
RPM = 21.2 g of Smartamine M
+ 38.8 g Dry Distillers Grain
or CON = 60 g Dry Distillers Grain
RPM
CON
From DIM 30 to DIM 126

32. 26 - 3319 - 26
Days in milk
34-60 Double Ovsynch 61-68
28 32 47 615 33
Days after AI
Ultrasound
videos
AI
Amniotic Vesicle and Embryo Size
Pregnancy diagnosis

33. Amniotic vesicle size
Day 33 n Volume (mm3
)
Primiparous
Control 31 610.6 ± 38.6
RPM 36 596.0 ± 36.9
P-value 0.71
Multiparous
Control 35 472.3 ± 28.6
RPM 45 592.1 ± 46.0
P-value 0.05
Ellipsoid
Volume

34. Embryo size
Ellipsoid
volume
Crown-rump length
Abdominal diameter
Day 33 n
Crown-Rump
Length (mm)
Abdominal
Diameter (mm)
Volume (mm3)
Primiparous
Control 35 10.4 ± 0.24 5.6 ± 0.11 169.6 ± 7.7
RPM 38 10.9 ± 0.24 5.7 ± 0.16 191.9 ± 14.3
P-value 0.10 0.54 0.21
Multiparous
Control 36 10.5 ± 0.24 5.3 ± 0.12 160.5 ± 9.2
RPM 44 11.0 ± 0.22 5.9 ± 0.18 209.3 ± 15.6
P-value 0.27 0.03 0.01

35. Primiparous Multiparous
CON RPM P CON RPM P
P/AI
at day 28
63.5
(40/63)
66.7
(42/63)
0.35 67.1
(51/76)
66.7
(54/81)
0.48
P/AI
at day 32
58.7
(37/63)
60.3
(38/63)
0.43 58.4
(45/77)
62.2
(51/82)
0.31
P/AI
at day 47
56.5
(35/62)
57.1
(36/63)
0.47 55.8
(43/77)
61.7
(50/81)
0.23
P/AI
at day 61
54.8
(34/62)
56.5
(35/62)
0.43 54.0
(41/76)
59.7
(46/77)
0.23
Preg. Loss
28-61
12.8
(5/39)
14.6
(6/41)
0.37 19.6
(10/51)
6.1
(3/49)
0.03
Preg. Loss
32-61
5.6
(2/36)
5.4
(2/37)
0.50 8.9
(4/45)
0.0
(0/46)
0.04
Fertility of Synchronized Cows

36. Critical Periods for Nutritional
Effects on Reproduction
Dry Period
(last 3 wks)
Early Post-
partum (3 wks)
Pre-AI
(1 wk)
Pregnancy
What is the role of nutrition in improving
reproductive efficiency in today’s dairy herds?
- Improve Nutrition in Dry Period – Vit E, lower energy
- Reduce Post-partum BCS loss
- Improve nutrition near AI – Lower NFC, Increase PUFA
- Optimize amino acids to reduce pregnancy loss