J. Dairy Sci.

101:7907–7929
https://doi.org/10.3168/jds.2018-14580
© American Dairy Science Association®, 2018.

Supplemental Table S1. Characteristics of cows enrolled in the experiment according to

treatment (mean ± SD)

Treatment1
Item Short -70 Short -180 Long -70 Long -180
Lactation prepartum
Mean 1.90 ± 0.94 1.83 ± 1.10 1.89 ± 0.96 2.00 ± 1.16
Primiparous, n 12 13 11 12
Multiparous, n 17 16 17 16
Body weight, kg 708.0 ± 86.5 745.2 ± 68.0 749.6 ± 86.0 724.1 ± 81.4
Body condition score, 1 to 5 3.64 ± 0.39 3.69 ± 0.36 3.63 ± 0.43 3.61 ± 0.33
Gestation length, d 277.5 ± 4.4 276.9 ± 4.7 275.8 ± 5.4 274.8 ± 5.1
Prepartum
Days dry 55.2 ± 16.0 55.1 ± 13.9 52.9 ± 8.3 53.1 ± 9.8
Days on experiment 44.1 ± 6.3 43.8 ± 6.1 44.1 ± 6.2 43.3 ± 6.1
Days fed negative DCAD 20.9 ± 4.3 20.4 ± 4.7 43.5 ± 6.0 43.0 ± 6.5
305-d milk, kg/d 34.0 ± 6.0 33.2 ± 4.9 32.3 ± 5.5 33.5 ± 4.5
Calf
Singleton female, n 11 8 10 14
Singleton male, n 16 19 15 14
Twin, n 2 2 3 0
1
From day -42 to -22 relative to calving, cows in treatments Short -70 and Short -180 were fed a diet with a DCAD of +110 mEq/kg, whereas cows in Long -70
were fed a containing -70 mEq/kg and cows in Long -180 were fed a diet containing -180 mEq/kg.

1
J. Dairy Sci. 101:7907–7929
https://doi.org/10.3168/jds.2018-14580
© American Dairy Science Association®, 2018.

Supplemental Table S2. Effects of level of dietary cation-anion difference (DCAD) on measures of energy status from d -42 to -22
relative to calving, blood acid-base status and concentrations of minerals in Holstein cows1
Treatment, DCAD as mEq/kg P-value2
Item +110 -70 -180 Treatment Linear Quadratic
Cows, n 58 28 28 --- --- ---
DM intake, kg/d 11.5 ± 0.3 10.7 ± 0.4 10.2 ± 0.4 0.01 0.02 0.55
Intake, Mcal NEL/d 16.7 ± 0.5 15.5 ± 0.6 14.8 ± 0.7 0.02 0.01 0.48
NEL balance, Mcal/d 1.2 ± 0.5 -0.1 ± 0.6 -0.5 ± 0.7 0.03 0.02 0.36
Body weight, Kg 752.8 ± 12.5 759.0 ± 15.5 734.8 ± 17.0 0.44 0.26 0.42
Body condition, 1 to 5 3.73 ± 0.06 3.67 ± 0.07 3.69 ± 0.08 0.65 0.66 0.49
Whole blood3
pH 7.426 ± 0.011 7.396 ± 0.015 7.397 ± 0.014 0.11 0.13 0.24
Total CO2, mM 27.8 ± 0.6 27.7 ± 0.8 24.6 ± 0.8 0.001 < 0.001 0.17
Saturation of O2, % 59.6 ± 2.8 58.6 ± 3.9 67.5 ± 3.9 0.15 0.06 0.34
pCO2, mm Hg 40.6 ± 1.0 43.4 ± 1.4 38.2 ± 1.4 0.02 0.07 0.02
pO2, mm Hg 31.6 ± 2.1 30.9 ± 2.9 35.8 ± 2.8 0.34 0.17 0.46
Bicarbonate, mM 26.6 ± 0.6 26.4 ± 0.8 23.6 ± 0.8 0.004 0.001 0.25
Base excess, mM 2.12 ± 0.69 1.59 ± 0.95 -1.17 ± 0.94 0.01 0.004 0.47
Ionized Ca, mM 1.245 ± 0.012 1.227 ± 0.017 1.248 ± 0.017 0.55 0.72 0.28
Na, mM 140.6 ± 0.3 140.7 ± 0.4 140.7 ± 0.4 0.96 0.88 0.85
K, mM 4.05 ± 0.08 4.11 ± 0.10 4.05 ± 0.10 0.83 0.89 0.54
Glucose, mM 3.65 ± 0.06 3.69 ± 0.09 3.75 ± 0.09 0.63 0.35 0.95
Urine pH 8.05 ± 0.06 6.52 ± 0.07 5.48 ± 0.08 < 0.001 < 0.001 < 0.001
1
From day -42 to -22 relative to calving, cows in treatments Short -70 and Short -180 were fed a diet with a DCAD of +110 mEq/kg, whereas cows in Long -70
were fed a containing -70 mEq/kg and cows in Long -180 were fed a diet containing -180 mEq/kg.
2
Treatment = effect of treatment (+110 vs. -70 vs. -180 mEq/kg); Linear = orthogonal polynomial contrast for the linear effect of DCAD; Quadratic = orthogonal
polynomial contrast for the quadratic effect of DCAD.
3
Analysis performed in jugular whole blood sampled at 250 d of gestation, approximately 25 d before calving. pO2 = partial pressure of O2; pCO2 = partial
pressure of CO2.

2
J. Dairy Sci. 101:7907–7929
https://doi.org/10.3168/jds.2018-14580
© American Dairy Science Association®, 2018.

Supplemental Table S3. Effects of duration (DUR) of prepartum feeding and level of dietary cation-anion difference (DCAD) on
measures of energy status in the last 21 d of gestation in Holstein cows1
Short Long P-value2
Item -70 -180 -70 -180 SEM DUR DCAD DUR x DCAD
DM intake, kg/d 11.1 9.7 10.5 9.6 0.5 0.38 0.005 0.45
Intake, Mcal NEL/d 16.0 14.0 15.1 13.9 0.7 0.43 0.007 0.51
NEL balance, Mcal/d 0.23 -2.16 -0.89 -1.90 0.72 0.49 0.007 0.27
Body weight
Kg 753.8ab 781.3a 771.6ab 747.5b 14.2 0.50 0.89 0.03
Change wk -6 to -1, kg/d 1.02a 0.44b 0.46b 0.70ab 0.20 0.37 0.29 0.01
Body condition
Score, 1 to 5 3.63 3.64 3.64 3.59 0.07 0.73 0.77 0.65
Change wk -6 to -1 -0.124 -0.178 -0.081 -0.221 0.063 0.99 0.06 0.40
1
Cows were fed the following diets according to treatment: Short -70 = diet containing +110 mEq/kg from 232 to 254 d of gestation, followed by a diet with -70
mEq/kg from 255 d of gestation to calving; Short -180 = diet containing +110 mEq/kg from 232 to 254 d of gestation, followed by a diet with -180 mEq/kg from
255 d of gestation to calving; Long -70 = diet containing -70 mEq/kg from 232 d of gestation to calving; Long -180 = diet containing -180 mEq/kg from 232 d of
gestation to calving.
2
DUR = effect of feeding duration (42 vs. 21 d); DCAD = effect of level of DCAD (-70 vs. -180); DUR x DCAD = effect of interaction between DUR and
DCAD.
a,b
Superscripts in the same row differ (P < 0.05) after adjustment by the method of Tukey.

3
J. Dairy Sci. 101:7907–7929
https://doi.org/10.3168/jds.2018-14580
© American Dairy Science Association®, 2018.

Supplemental Table S4. Effects of duration (DUR) of prepartum feeding and level of dietary cation-anion difference (DCAD) on
blood concentrations of minerals and metabolites in Holstein cows prepartum1
Short Long P-value2
Item -70 -180 -70 -180 SEM DUR DCAD DUR x DCAD
Blood iCa, mM3 1.236 1.272 1.230 1.271 0.013 0.78 0.002 0.83
Plasma tCa, mM4 2.41 2.40 2.39 2.36 0.03 0.13 0.34 0.73
Plasma tMg, mM4 0.75 0.72 0.75 0.76 0.01 0.25 0.66 0.27
Plasma tP, mM4 1.74b 1.82ab 1.83a 1.77ab 0.05 0.52 0.66 0.008
Blood Na, mM3 143.0 143.6 143.2 143.5 0.4 0.99 0.27 0.77
Blood K, mM3 4.00 4.03 4.15 4.13 0.09 0.12 0.95 0.76
Plasma glucose, mM4 3.58 3.59 3.62 3.64 0.06 0.88 0.65 0.11
Plasma NEFA, mM4 0.260ab 0.314a 0.275ab 0.239b 0.027 0.20 0.76 0.05
Plasma BHB, mM4 0.693 0.732 0.699 0.666 0.030 0.21 0.95 0.15
1
Cows were fed the following diets according to treatment: Short -70 = diet containing +110 mEq/kg from 232 to 254 d of gestation, followed by a diet with -70
mEq/kg from 255 d of gestation to calving; Short -180 = diet containing +110 mEq/kg from 232 to 254 d of gestation, followed by a diet with -180 mEq/kg from
255 d of gestation to calving; Long -70 = diet containing -70 mEq/kg from 232 d of gestation to calving; Long -180 = diet containing -180 mEq/kg from 232 d of
gestation to calving.
2
DUR = effect of feeding duration (42 vs. 21 d); DCAD = effect of level of DCAD (-70 vs. -180); DUR x DCAD = effect of interaction between DUR and DCAD
(Short -70 plus Long -180 vs. Short -180 plus Long -70).
3
Jugular blood sampled on d -10 and -6 relative to calving and analyzed for concentrations of iCa, Na, and K.
4
Blood sampled by puncture of coccygeal vessels on d -14, -12, -10, -8, -6, -4, and -2 relative to calving and plasma analyzed for concentrations of tCa, tMg, tP,
glucose, NEFA, and BHB.
a,b
Superscripts in the same row differ (P < 0.05) after adjustment by the method of Tukey.

4
J. Dairy Sci. 101:7907–7929
https://doi.org/10.3168/jds.2018-14580
© American Dairy Science Association®, 2018.

Supplemental Table S5. Effects of duration (DUR) of prepartum feeding and level of dietary cation-anion difference (DCAD) on
measures of acid-base balance in Holstein cows postpartum1
Short Long P-value2
Item3 -70 -180 -70 -180 SEM DUR DCAD DUR x DCAD
Blood pH 7.451 7.456 7.446 7.455 0.004 0.50 0.09 0.59
Total CO2, mM 30.4 30.6 29.8 31.2 0.4 0.90 0.05 0.12
Saturation of O2, % 61.4 65.3 63.0 62.4 1.3 0.59 0.20 0.08
PCO2, mm Hg 41.6 41.4 41.3 42.2 0.6 0.66 0.50 0.37
PO2, mm Hg 31.2 32.5 32.0 31.5 0.6 0.88 0.43 0.11
Bicarbonate, mM 29.2ab 29.3ab 28.6b 30.0a 0.4 0.94 0.04 0.08
Base excess, mM 5.17 5.52 4.56 6.01 0.41 0.87 0.02 0.15
1
Cows were fed the following diets according to treatment: Short -70 = diet containing +110 mEq/kg from 232 to 254 d of gestation, followed by a diet with -70
mEq/kg from 255 d of gestation to calving; Short -180 = diet containing +110 mEq/kg from 232 to 254 d of gestation, followed by a diet with -180 mEq/kg from
255 d of gestation to calving; Long -70 = diet containing -70 mEq/kg from 232 d of gestation to calving; Long -180 = diet containing -180 mEq/kg from 232 d of
gestation to calving.
2
DUR = effect of feeding duration (42 vs. 21 d); DCAD = effect of level of DCAD (-70 vs. -180); DUR x DCAD = effect of interaction between DUR and DCAD
(Short -70 plus Long -180 vs. Short -180 plus Long -70).
3
Jugular blood sampled and analyzed on d 0, 1, 2, 3, 4 postpartum.
a,b
Superscripts in the same row differ (P < 0.05) after adjustment by the method of Tukey.