Response of lactating dairy cows to high protein distillers grains or 3 other protein supplements.

Abstract

This study compared high protein dried distillers grains (HPDDG) with soybean meal (SBM), canola meal (CM), and dried distillers grains with solubles (DDGS) as protein supplements in dairy diets. A lactation trial used 12 multiparous cows averaging 78 d in milk at the start of the experiment in a 4 x 4 Latin square design with 28-d periods. Weeks 1 and 2 of each period were used for adjustment and wk 3 and 4 for data collection. Each treatment diet consisted of 55% forage and one of the 4 protein supplements in a concentrate mix. Total mixed diets averaged 15.3% crude protein, with 38% of the protein from one of the 4 protein supplements. Dry matter intake (24.4 kg/d) and crude protein intake (3.57 kg/d) were similar for all 4 diets. Milk production (31.8 kg/d), protein yield (1.05 kg/d), fat yield (1.29 kg/d), and protein percentage (3.31) were similar for all 4 treatment diets. Milk fat percentage was lower when fed DDGS (3.78) than when fed SBM or HPDDG (4.21), but similar with CM (4.07). Feed efficiency (1.44kg of energy-corrected milk/kg of dry matter intake) and nitrogen efficiency (0.29) were not affected by diet. Total milk nitrogen and true milk protein were highest when fed the HPDDG diet. Molar proportions of acetate, propionate, and the acetate to propionate ratio in ruminal contents and ruminal ammonia concentrations were similar for all diets. Arterial and venous concentrations of total essential AA tended to be lower when fed CM, reflecting lower concentrations of His, Ile, Leu, and Val when fed the CM diet. Extraction efficiency of AA from blood by the mammary gland indicated that Met was the first limiting AA when fed the SBM diet, whereas Lys was first limiting for the other diets. Phenylalanine was third limiting with all diets. Feeding HPDDG was equally as effective as feeding SBM, CM, and regular distillers grains as a protein supplement for lactating cows.

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Response of lactating dairy cows to high protein distillers grains
or 3 other protein supplements
K. A. Christen ,*1 D. J. Schingoethe ,*2 K. F. Kalscheur ,* A. R. Hippen ,* K. K. Karges ,† and M. L. Gibson †
* Dairy Science Department, South Dakota State University, Brookings 57007-0647
† Dakota Gold Research Association, Sioux Falls, SD 57104
ABSTRACT
This study compared high protein dried distillers
grains (HPDDG) with soybean meal (SBM), canola meal
(CM), and dried distillers grains with solubles (DDGS)
as protein supplements in dairy diets. A lactation trial
used 12 multiparous cows averaging 78 d in milk at the
start of the experiment in a 4 × 4 Latin square design
with 28-d periods. Weeks 1 and 2 of each period were
used for adjustment and wk 3 and 4 for data collection.
Each treatment diet consisted of 55% forage and one of
the 4 protein supplements in a concentrate mix. Total
mixed diets averaged 15.3% crude protein, with 38%
of the protein from one of the 4 protein supplements.
Dry matter intake (24.4 kg/d) and crude protein intake
(3.57 kg/d) were similar for all 4 diets. Milk production
(31.8 kg/d), protein yield (1.05 kg/d), fat yield (1.29
kg/d), and protein percentage (3.31) were similar for
all 4 treatment diets. Milk fat percentage was lower
when fed DDGS (3.78) than when fed SBM or HPDDG
(4.21), but similar with CM (4.07). Feed efficiency (1.44
kg of energy-corrected milk/kg of dry matter intake)
and nitrogen efficiency (0.29) were not affected by diet.
Total milk nitrogen and true milk protein were highest
when fed the HPDDG diet. Molar proportions of ace-
tate, propionate, and the acetate to propionate ratio in
ruminal contents and ruminal ammonia concentrations
were similar for all diets. Arterial and venous concen-
trations of total essential AA tended to be lower when
fed CM, reflecting lower concentrations of His, Ile, Leu,
and Val when fed the CM diet. Extraction efficiency
of AA from blood by the mammary gland indicated
that Met was the first limiting AA when fed the SBM
diet, whereas Lys was first limiting for the other diets.
Phenylalanine was third limiting with all diets. Feeding
HPDDG was equally as effective as feeding SBM, CM,
and regular distillers grains as a protein supplement for
lactating cows.
Key words: distillers grain , high protein distillers
grain , canola meal , lactating cow
INTRODUCTION
Dried distillers grains with solubles (DDGS) is the
primary coproduct of ethanol production from corn
and other grains. Several studies (e.g., Nichols et al.,
1998; Anderson et al., 2006; Kleinschmit et al., 2006)
demonstrated that DDGS can effectively replace soy-
bean meal (SBM) as a protein source, but there is less
evidence comparing DDGS with other protein supple-
ments. Mulrooney et al. (2009) reported similar milk
production when fed DDGS or canola meal (CM) as
the protein supplement, although milk yield and pro-
tein yield responses and extraction efficiency of AA by
the mammary gland (i.e., arteriovenous difference of
an AA across the mammary gland relative to arterial
concentration of the AA) indicated some benefits of
feeding a combination of CM and DDGS rather than
only one of them as the protein supplement.
Ethanol plants are attempting to improve the fer-
mentation of corn to ethanol and this is resulting in
new products (Robinson et al., 2008). One way this is
accomplished is through fractionation of the corn in
which the germ and the bran are removed before fer-
mentation. This new product, called high protein dried
distiller grains (HPDDG), is higher in CP and lower
in fat (44% CP, 3.4% fat) than DDGS (30% CP, 10%
fat) produced through traditional ethanol fermentation
processes (Robinson et al., 2008). The HPDDG is a
product that is closer in nutrient composition to CM
and SBM. Hubbard et al. (2009) recently reported that
feeding HPDDG increased milk production relative to
the feeding of an SBM-based diet, but HPDDG has not
been compared with CM. The objective of this study
was to determine the response in milk production to
diets that contained each of SBM, HPDDG, CM, or
DDGS as the supplemental protein.
MATERIALS AND METHODS
All procedures for this study were conducted under
approval of the South Dakota State University Animal
J. Dairy Sci. 93 :2095–2104
doi: 10.3168/jds.2009-2687
© American Dairy Science Association®, 2010 .
2095
Received September 1, 2009.
Accepted February 2, 2010.
1 Current address: Vita Plus Corp., Loyal, WI 54446.
2 Corresponding author: david.schingoethe@sdstate.edu

Page 2

Care and Use Committee. Twelve multiparous Holstein
cows (78 ± 24 DIM) were used in a replicated 4 × 4
Latin square design with 28-d periods. Weeks 1 and 2
of each period were for adjustment to diets and wk 3
and 4 for data collection. Dietary treatments consisted
of 4 different protein sources (SBM, HPDDG, CM, and
DDGS) intended to supply equal amounts of protein
(approximately 38% of the total dietary protein) from
the respective supplemental protein sources. All diets
contained 27.5% alfalfa hay and 27.5% corn silage on
a DM basis (Table 1). The diets were formulated to be
isonitrogenous at 16.0% CP and isolipogenic at 4.7% fat
(Table 2). Energy Booster (Milk Specialties Company,
Dundee, IL) was added to all except the DDGS diet to
maintain approximately equal ether extract concentra-
tion in all diets. All other nutrients were formulated
to meet or exceed the requirements according to NRC
(2001). Cows were housed in a free-stall barn and in-
dividually fed diets as a TMR for ad libitum intake
once daily (0800 h) using Calan Broadbent feeder doors
(American Calan Inc., Northwood, NH). Feed intakes
were recorded daily.
Samples of forages, concentrates, SBM, HPDDG, CM,
and DDGS were collected for 2 consecutive days at the
end of each period and stored at −20°C until analysis.
Samples were composited by period and dried at 55°C
in a Despatch oven (Despatch Oven Co., Minneapo-
lis, MN) for 48 h. Composites were ground through a
4-mm screen in a Wiley mill (Arthur H, Thomas Co.,
Philadelphia, PA) and then through a 1-mm screen in
an ultracentrifuge mill (Brinkman Instruments Co.,
Westbury NY). Samples were corrected to 100% DM
by drying an aliquot of the composite at 105°C for 24 h
(method 930.15; AOAC, 2002). Samples were analyzed
for CP (method 990.03; AOAC, 2002), ether extract
with petroleum ether (method 930.39; AOAC, 2002),
and ash (method 942.05; AOAC, 2002). Neutral de-
tergent fiber was determined with sodium sulfite and
α-amylase (Van Soest et al., 1991), and ADF (Robert-
son and Van Soest, 1981) sequentially with an Ankom
Fiber Analyzer (Ankom Technology Corp., Fairport,
NY). Composition of the TMR was calculated based on
analyses and concentrations in the diets of the concen-
trate mix and the individual forages.
Cows were milked 3 times daily at 0600, 1400, and
2100 h and yields were recorded. Milk samples were
collected for each milking on 2 consecutive days during
the last 2 wk of each period. Samples were mixed by
gentle inversion and composited by weight correspond-
ing to the respective milking for each cow on sampling
day. These samples were sent to Heart of America DHI
laboratory (Manhattan, KS) for composition analysis
where fat, true protein, and lactose were determined
using mid-infrared spectroscopy (Bentley 2000 Infra-
red Milk Analyzer, Bentley Instruments, Chaska, MN;
method 972.16; AOAC, 2002). Concentrations of MUN
were determined using chemical methodology based on
a modified Berthelot reaction (ChemSpec 150 Analyz-
er, Bentley Instruments; Chaney and Marbach, 1962),
and SCC was determined using a flow cytometer laser
Journal of Dairy Science Vol. 93 No. 5, 2010
CHRISTEN ET AL.
2096
Table 1. Ingredient composition of diets1
Item, % of DM SBMHPDDGCM DDGS
Corn silage
Alfalfa hay
Corn ground dry
DDGS
SBM, 44% CP
HPDDG
Canola meal
Fat2
Dicalcium phosphate
Magnesium oxide
Limestone
Salt
Mineral–vitamin mix3
Vitamin A, D, E mix4
Vitamin E5
27.5
27.5
29.3
0.0
11.1
0.0
0.0
2.0
0.40
0.07
0.73
0.55
0.6
0.2
0.05
27.5
27.5
28.4
0.0
0.0
12.0
0.0
1.8
0.60
0.08
0.72
0.55
0.6
0.2
0.05
27.5
27.5
28.8
0.0
0.0
0.0
12.7
1.2
0.17
0.05
0.71
0.55
0.6
0.2
0.05
27.5
27.5
21.3
21.2
0.0
0.0
0.0
0.0
0.0
0.07
1.05
0.55
0.6
0.2
0.05
1SBM = soybean meal; HPDDG = high protein dried distillers grains with solubles; CM = canola meal;
DDGS = dried distillers grains with solubles. Dietary treatments consisted of 4 different protein sources (SBM,
HPDDG, CM, and DDGS) intended to supply equal amounts of protein (approximately 38% of the total di-
etary protein) from the respective supplemental protein sources.
2Energy Booster 100 (Milk Specialties Company, Dundee, IL).
3Guaranteed analysis: Mg, 10.0%; Mn, 1.7%; Zn, 2.6%; Cu, 4,712 mg/kg; Co, 119 mg/kg; I, 396 mg/kg; Se, 140
mg/kg; Fe, 4,640 mg/kg; vitamin A, 2,640,000 IU/kg; vitamin D, 528,000 IU/kg; vitamin E, 10,560 IU/kg.
4Guaranteed analysis: vitamin A, 5,500,000 IU/kg; vitamin D, 880,000 IU/kg; vitamin E, 2,200 IU/kg.
544,000 IU/kg.

Page 3

(Somacount 500, Bentley Instruments; method 975.16;
AOAC, 2002). An extra sample was taken one day
during the last week of each period for milk nitrogen
fractions. Samples were mixed by gentle inversion and
analyzed for total milk protein (method 991.20; AOAC,
2002), nonprotein nitrogen (method 991.21; AOAC,
2002), and noncasein nitrogen (method 998.05; AOAC,
2002), with true protein (method 991.23; AOAC, 2002)
and casein nitrogen (method 998.07; AOAC, 2002) cal-
culated by difference.
Body condition scores on a scale of 1 to 5 (Wildman
et al., 1982) and BW were recorded approximately 3 h
after feeding for 3 consecutive days at the start of the
experiment and at the end of each period. Blood was
collected at the end of each period from the coccygeal
artery and subcutaneous abdominal vein into vacuum
tubes containing heparin (Becton Dickinson and Co.,
Franklin Lakes, NJ). Plasma was obtained by centrifug-
ing at 500 × g and stored at −20°C until analyzed for
AA via HPLC (model 1100, Agilent Technologies Inc.,
Palo Alto, CA) with a PCX 5200 postcolumn deriva-
tizer (Pickering Laboratories Inc., Mountain View, CA)
as described by Mondina et al. (1972), Pickering (1989),
and Grunau and Swiader (1992). Extraction efficiencies
of AA by the mammary gland were calculated based
on the following equation: arteriovenous difference of
AA (g/L) × 100/concentration (g/L) in plasma of coc-
cygeal artery.
Samples of rumen fluid were obtained via an esopha-
geal tube fitted with a suction strainer on 2 consecutive
days at the end of each period approximately 3 h after
feeding. The first 150 mL was discarded to minimize
saliva contamination and then approximately 250 mL
of rumen fluid was collected. A 10-mL aliquot was
mixed with 2 mL of 25% (wt/vol) metaphosphoric acid
and frozen at −20°C until centrifuged at 20,000 × g
and analyzed for concentrations of ammonia-N (Weath-
erburn, 1976) and VFA. Concentrations of VFA were
2097
LACTATION RESPONSE TO 4 PROTEIN SUPPLEMENTS
Journal of Dairy Science Vol. 93 No. 5, 2010
Table 2. Nutrient composition of diets1
Item SBMHPDDG CMDDGS
Formulated, % of DM unless noted
CP
RDP2
RUP2
Ether extract
ADF
NDF
Forage NDF
NFC
NEL,2 Mcal/kg
Ca
P
Measured, % of DM unless noted
DM, %
CP
Ether extract
ADF
NDF
Forage NDF
Ash
NFC3
NEL,2 Mcal/kg
Ca
P
Mg
K
S
PSPS DM basis, mm4
>19.0
19.0–8.0
8.0–1.18
<1.18

16.0
11.1
4.9
4.7
19.0
27.3
22.9
46.4
1.59
0.9
0.4

68.8
15.4
4.7
16.2
27.4
22.7
6.22
42.4
1.54
1.01
0.33
0.35
1.00
0.19

15.3
21.7
38.3
24.7
16.0
10.2
5.8
4.7
19.2
28.0
22.9
47.0
1.59
0.9
0.4

68.8
15.0
4.6
16.6
31.1
22.7
6.27
40.5
1.56
0.89
0.32
0.33
0.89
0.21

14.9
21.5
36.4
27.2
16.0
11.0
5.0
4.7
20.5
29.4
22.9
45.1
1.59
0.9
0.4

69.6
15.6
4.5
17.7
28.9
22.7
6.71
41.5
1.56
1.05
0.35
0.35
0.93
0.22

16.2
21.8
34.9
27.1
16.0
10.1
5.8
4.7
19.6
30.3
22.9
45.1
1.59
0.9
0.4

69.2
15.3
4.1
17.0
31.2
22.7
6.22
40.8
1.58
1.00
0.35
0.34
0.97
0.26

17.3
21.4
28.4
32.0
1SBM = soybean meal; HPDDG = high protein dried distillers grains with solubles; CM = canola meal;
DDGS = dried distillers grains with solubles. Dietary treatments consisted of 4 different protein sources (SBM,
HPDDG, CM, and DDGS) intended to supply equal amounts of protein (approximately 38% of the total di-
etary protein) from the respective supplemental protein sources.
2Estimated values from NRC (2001).
3NFC = 100 − ether extract − CP − NDF − ash.
4PSPS = Penn State Particle Separator.

Page 4

measured using a 6890 gas chromatograph (Hewlett-
Packard, Anondale, PA) equipped with a column (0.25
mm i.d. × 15 m; Nukol, 17926 to 01C, Supelco Inc.,
Bellenfonte, PA). The split ratio in the injector port
(250°C) was 100:1 with a flow of 1.3 mL/min of He.
Column and detector temperatures were maintained at
130 and 225°C, respectively.
Means of DMI, milk yield, and milk composition were
used in statistical analysis. Analysis of variance was
conducted using the MIXED procedure of SAS (SAS
Institute Inc., Cary, NC): Y = treatment + square +
period + cow (square), where cow (square) was the
random variable. The treatment × square and treat-
ment × period interactions were tested and removed
because they were not significant. Least squares means
of the treatments were calculated and separation was
determined using PDIFF. Significance was declared at
P < 0.05 and tendencies noted at P < 0.10.
RESULTS AND DISCUSSION
Nutrient Composition
All diets (Table 2) were slightly lower in CP (15.3%)
than the anticipated 16% CP, which was acceptable
because with a protein evaluation study such as this
it was important that diets not contain excess protein.
Analyses of ingredients (Table 3) indicated where these
minor discrepancies occurred. The alfalfa hay contained
less protein than preliminary analyses indicated. Also,
the CP in the HPDDG (44.6%) was lower than values
used in formulation (46%). The ether extract content of
the DDGS used was on the low end of the fat content of
typical DDGS, which is why the DDGS diet contained
slightly less ether extract (4.1%) than the other diets
(4.6%).
Particle Size
Particle size of diets was determined on all treatment
diets using the Penn State Particle Separator during
wk 3 and 4 of each period (Table 2; Kononoff et al.,
2003). Each sieve was dried to determine the percent
on a DM basis, as recommended by Kononoff et al.
(2003). The amount of DM retained on the top sieve of
the particle separator was between 14.9 and 17.3%. The
middle sieve was low in all diets, ranging from 21.4 to
21.8%. The screen and pan had from 28.4 to 38.3% on
the screen and 24.7 to 32.0% in the pan. All diets were
similar in particle size distribution and within normal
expected ranges.
DMI, Milk Production, and BW
Dry matter intake (24.8 kg/d), milk production (31.8
kg/d), ECM (34.7 kg/d), and feed efficiency (1.44)
were similar for all diets (Table 4). This was similar to
results by Liu et al. (2000), where there was no change
in milk production when feeding DDGS versus a blend
of SBM, DDGS, and fishmeal. However, Nichols et al.
(1998) and Anderson et al. (2006) observed an increase
in milk production when feeding DDGS versus SBM-
based control diets. But, those studies did not balance
the fat content of all diets; thus, part of their response
might have been attributable to the increased fat con-
tent of the DDGS diets. Hubbard et al. (2009) observed
increased milk production when HPDDG was fed in
place of SBM. Piepenbrink et al. (1998), Brito and
Journal of Dairy Science Vol. 93 No. 5, 2010
CHRISTEN ET AL.
2098
Table 3. Composition of forages and concentrate mixes for treatment diets1
Item, % of DM unless noted
Ingredient Concentrate mix
AlfalfaCorn silage SBMHPDDG CM DDGSCornSBMHPDDG CM DDGS
DM, %
CP
EE2
ADF
NDF
NFC3
Ash
Ca
Mg
P
K
S
88.6
19.5
1.41
32.1
42.6
27.1
9.41
1.59
0.38
0.27
1.80
0.23
47.4
7.00
2.50
21.2
40.1
46.3
4.05
0.20
0.25
0.22
0.60
0.11
88.0
50.6
1.49
5.55
9.08
31.5
7.35
0.56
0.30
0.71
2.46
0.42
90.4
44.5
3.42
10.1
28.7
21.9
1.93
0.01
0.11
0.43
0.39
0.86
92.4
43.7
3.77
16.4
22.0
21.4
9.13
1.44
0.59
1.16
1.38
0.84
89.9
29.8
9.57
5.18
26.1
29.5
5.06
0.04
0.37
0.89
1.18
1.02
86.2
8.44
2.30
2.37
9.66
78.6
1.00





87.7
18.1
4.72
3.44
10.4
59.0
7.81





88.2
17.2
4.20
4.21
18.6
52.5
7.52





88.5
18.4
4.17
6.64
13.8
55.5
8.11





88.6
17.9
3.40
5.18
18.8
51.6
8.31





1SBM = soybean meal; HPDDG = high protein dried distillers grains with solubles; CM = canola meal; DDGS = dried distillers grains with
solubles. Dietary treatments consisted of 4 different protein sources (SBM, HPDDG, CM, and DDGS) intended to supply equal amounts of
protein (approximately 38% of the total dietary protein) from the respective supplemental protein sources.
2EE = ether extract.
3NFC = 100 − EE − CP − NDF − ash.

Page 5

Broderick (2007), and Sanchez and Claypool (1983) all
observed similar milk production when feeding CM in
place of SBM; however, Brito and Broderick (2007) and
Sanchez and Claypool (1983) reported an increase in
DMI when CM was fed in place of SBM. Mulrooney et
al. (2009) observed similar production when DDGS or
CM was fed and slightly higher production with blends
of the 2 protein supplements. Body weights (684 kg;
Table 4), BW changes, BCS (3.47), and BCS changes
were similar across treatments. No treatment effects on
these parameters were expected.
Milk Composition
Milk fat percentage (Table 4) was higher with the
SBM and HPDDG diets than with the DDGS diet,
whereas with the CM diet it was similar to all other
diets. The lower fat content observed when DDGS was
fed with adequate forage fiber as in this experiment
was somewhat unexpected because Kalscheur (2005)
observed that milk fat content was lower only in diets
supplemented with wet or dried distillers grains with
solubles diets that contained less than 50% forage and
22% forage NDF. The similar milk fat percentage with
the SBM and CM diets agreed with others (Sanchez
and Claypool, 1983; Piepenbrink et al., 1998; Brito and
Broderick, 2007; Mulrooney et al., 2009). When Hub-
bard et al. (2009) fed HPDDG in place of SBM, milk
fat yields were higher because of higher milk production
coupled with numerically higher fat test.
Milk protein content was higher in the SBM, HP-
DDG, and CM diets than in the DDGS diet, but pro-
tein yield was similar across all diets because of the
numerically higher milk yield when DDGS was fed.
Milk protein content is seldom affected by feeding
distillers grains with solubles unless protein is limit-
ing in the diet (Schingoethe et al., 2009), which may
have occurred in this study because the diets contained
marginal amounts of protein, especially RDP (NRC,
2001). Then the Lys limitation in DDGS may cause a
slight decrease in milk protein content (Nichols et al.,
1998; Kleinschmit et al., 2006). Considering the limita-
tions of calculating estimated balances of RDP (NRC,
2001), all 4 diets were low but positive for estimated
RDP balance (359, 104, 247, and 133g/d for the SBM,
HPDDG, CM, and DDGS diets, respectively), with the
2 distillers product diets having the lowest RDP (Table
2) and RDP balance. Estimated balances for RUP (−8,
208, 9, and 87 g/d, respectively) are even less reliable
because accurate data are not available on HPDDG
or on some of the other dietary ingredients. Thus, the
above-estimated RDP and RUP balances illustrate that
2099
LACTATION RESPONSE TO 4 PROTEIN SUPPLEMENTS
Journal of Dairy Science Vol. 93 No. 5, 2010
Table 4. Dry matter intake, milk yield, milk composition, and BW (LSM) from lactating dairy cattle fed
treatment diets1
Item
Diet2
SEMP-value SBMHPDDGCM DDGS
DMI, kg/d
Milk yield, kg/d
ECM,3 kg/d
ECM/DMI
N efficiency4
Fat, %
Fat, kg/d
Protein, %
Protein, kg/d
Lactose, %
Lactose, kg/d
MUN, mg/dL
SCC, 103 cells/mL
BW, kg
BW, kg of change/d
BCS5
BCS, change/d
24.1
31.7
35.1
1.48
0.299
4.21a
1.33
3.33a
1.04
4.71
1.50
9.85
535
683
−0.65
3.45
0.0004
24.6
31.2
34.8
1.45
0.300
4.21a
1.31
3.36a
1.05
4.73
1.48
9.21
359
681
−0.93
3.47
0.07
25.2
31.7
34.6
1.38
0.272
4.07ab
1.28
3.33a
1.05
4.72
1.50
9.83
359
686
−1.04
3.47
0.03
23.6
32.7
34.3
1.46
0.296
3.78b
1.24
3.23b
1.05
4.75
1.55
9.81
324
684
−0.88
3.47
0.06
0.85
1.59
1.74
0.09
0.02
0.16
0.08
0.09
0.05
0.06
0.09
0.45
0.45
0.63
0.95
0.64
0.29
0.02
0.53
0.08
0.99
0.58
0.65
0.50
0.21
0.56
0.11
0.87
0.35
211
17.9
0.14
0.08
0.03
a,bValues within rows with different superscripts differ (P < 0.05).
1Least squares means.
2SBM = soybean meal; HPDDG = high protein dried distillers grains with solubles; CM = canola meal;
DDGS = dried distillers grains with solubles. Dietary treatments consisted of 4 different protein sources (SBM,
HPDDG, CM, and DDGS) intended to supply equal amounts of protein (approximately 38% of the total di-
etary protein) from the respective supplemental protein sources.
3ECM = 0.327 × milk (kg) + 12.95 × fat (kg) + 7.20 × protein (kg).
4N in milk/N intake.
5BCS on scale of 1 to 5, where 1 = emaciated and 5 = overly fat (Wildman et al., 1982).

Page 6

using such simulated calculations based on currently
available NRC (2001) formulations can be inaccurate
and possibly of minimal value. Sanchez and Claypool
(1983) and Brito and Broderick (2007) observed no
change in milk protein percentages when feeding CM,
whereas Piepenbrink et al. (1998) observed an increase
in protein percentage when CM was fed and Mulrooney
et al. (2009) observed similar milk protein percentages
when fed DDGS, CM, or their blends. Milk urea nitro-
gen concentrations were similar across all treatments
but were low, likely because of the intended low CP in
the diets.
Milk Protein Fractions
Milk protein fractions (Table 5) were analyzed to
determine whether the AA profile of the protein source
fed affected milk protein. Total and true protein con-
tents were highest (P < 0.05) with the HPDDG diets at
3.43 and 3.28%, respectively. When fed the SBM, CM,
and DDGS diets, total and true protein contents were
similar (P > 0.05). The percentage of casein in the milk
was highest with HPDDG and lowest with the SBM
and DDGS diets (P < 0.05). With CM, casein content
was in the middle, which was similar (P > 0.05) to
the content with all other diets. This showed that the
increase in true protein could be partly attributable to
an increase in casein.
Rumen Parameters
The total VFA concentrations and proportions of
various VFA (Table 6) were not affected by the dietary
treatment (P > 0.07). This is in contrast with Nich-
ols et al. (1998), where the addition of DDGS to the
diet resulted in a decrease in total VFA. Piepenbrink
et al. (1998) observed decreased total VFA concentra-
tions when CM was included in the diet. The propor-
tions of acetate (68.3%), propionate (18.2%), butyrate
(9.99%), and isovalerate (1.37%) were not affected by
treatments, which indicated a similar fermentation pat-
tern among diets and was consistent with other studies
(Nichols et al., 1998; Anderson et al., 2006; Brito and
Broderick, 2007). There was a trend (P > 0.07) for
slightly lower concentrations of isobutyrate and valer-
ate with HPDDG and CM, but these changes did not
necessarily reflect the branched-chain AA composition
of the protein supplements (NRC, 2001), were not likely
biologically significant, and did not relate to RDP bal-
ance or AA composition of the protein supplements.
Ammonia concentrations in the rumen (3.9 mg/dL)
were unaffected by treatments but were relatively low,
as expected, because diets were low in protein (Roffler
and Satter, 1975).
Blood AA
Total essential AA concentrations in arterial plasma
(Table 7) tended to be lower when fed CM and con-
centrations in venous plasma were lower (P < 0.05)
with CM (Table 8), as also reported by Piepenbrink
and Schingoethe (1998). Leucine and Phe concentra-
tions were higher (P < 0.05) with HPDDG than with
SBM and CM and tended (P > 0.05) to be higher with
DDGS. There was a tendency for an increase in venous
concentrations of Met when HPDDG was fed as com-
pared with SBM, which was somewhat related to the
higher Met concentration in corn protein. There was no
effect of treatment on the venous or arterial concentra-
tions of Lys in this experiment. As expected based on
the AA composition of the protein supplements (NRC,
2001), Met concentration in plasma tended to be lowest
with the SBM diet whereas Lys tended to be lowest
with the 2 distillers product diets, which again reflected
Journal of Dairy Science Vol. 93 No. 5, 2010
CHRISTEN ET AL.
2100
Table 5. Protein fractionations of milk protein (LSM) from lactating dairy cattle fed treatment diets
Item
Diet1
SEM P-valueSBMHPDDG CMDDGS
Total CP,2 %
NPN,2 %
True protein,2 %
Casein N,2 %
% of total protein
True
Casein
3.36b
0.20
3.17b
2.65b

93.9
78.9
3.43a
0.19
3.28a
2.77a

94.4
79.2
3.33b
0.19
3.15b
2.67ab

93.3
79.7
3.29b
0.19
3.09b
2.57b

94.1
78.3
0.09
0.01
0.09
0.08

0.29
0.81
0.01
0.25
0.01
0.01

0.17
0.41
a,bValues within rows with different superscripts differ (P < 0.05).
1SBM = soybean meal; HPDDG = high protein dried distillers grains with solubles; CM = canola meal;
DDGS = dried distillers grains with solubles. Dietary treatments consisted of 4 different protein sources (SBM,
HPDDG, CM, and DDGS) intended to supply equal amounts of protein (approximately 38% of the total di-
etary protein) from the respective supplemental protein sources.
2N × 6.38.

Page 7

AA composition of the protein supplements. Arterio-
venous differences (Table 9) were essentially similar for
all diets.
Transfer efficiency and uptake to output ratios
consider only AA relative to secretion as milk pro-
tein (Kleinschmit et al., 2007). Extraction efficiency,
however, may be the ideal method for evaluating AA
requirements because it takes into account the entire
essential AA needs of the mammary gland and includes
AA extracted for all needs such as protein synthesis
and catabolism (Schingoethe, 1996; Nichols et al., 1998;
Kleinschmit et al., 2007). Also, the possibility of inac-
curacies when estimating mammary blood flow is not
a concern because mammary blood flow is not a part
of the calculations for extraction efficiency, whereas it
is included in the other 2 calculation methods. In this
2101
LACTATION RESPONSE TO 4 PROTEIN SUPPLEMENTS
Journal of Dairy Science Vol. 93 No. 5, 2010
Table 6. Ruminal VFA and ammonia (LSM) from lactating dairy cattle fed treatment diets1
Measurement
Diet
SEMP-valueSBM HPDDG CMDDGS
VFA, %
Acetate (A)
Propionate (P)
Isobutyrate
Butyrate
Isovalerate
Valerate
A:P
Total, mM/L
Ammonia, mg/dL

67.8
18.3
1.07a
10.2
1.44
1.20ab
3.74
49.7
4.50
68.9
17.8
0.90b
9.68
1.43
1.16b
3.95
52.6
3.93
69.0
18.0
0.92b
9.58
1.34
1.18b
3.87
54.8
4.49
67.4
18.6
0.95ab
10.5
1.28
1.33a
3.66
43.2
2.52
0.77
0.57
0.06
0.36
0.09
0.05
0.16
6.35
0.83
0.33
0.69
0.07
0.17
0.42
0.08
0.52
0.48
0.18
a,bValues within rows with different superscripts differ (P < 0.05).
1SBM = soybean meal; HPDDG = high protein dried distillers grains with solubles; CM = canola meal;
DDGS = dried distillers grains with solubles. Dietary treatments consisted of 4 different protein sources (SBM,
HPDDG, CM, and DDGS) intended to supply equal amounts of protein (approximately 38% of the total di-
etary protein) from the respective supplemental protein sources.
Table 7. Amino acid concentrations in plasma from the coccygeal artery (LSM) in lactating dairy cows fed
treatment diets1
Item
Diet
SEMP-value SBMHPDDG CMDDGS
Essential AA, μmol/dL
Arg
His
Ile
Leu
Lys
Met
Phe
Thr
Trp
Val
Total essential AA
Nonessential AA, μmol/dL
Ala
Asp
Asn
Glu
Gln
Gly
Pro
Ser
Tyr
Total nonessential AA

8.73
4.87a
11.90
15.37bc
8.01
1.83
3.65b
8.28
3.01
25.68a
91.27a

20.87
3.18
0.73
24.24
4.29b
27.04
6.26b
8.64
4.26b
105.53
7.56
5.31a
10.69
18.54a
7.50
2.18
4.40a
9.41
3.16
24.20a
92.98a

23.01
2.87
0.86
27.59
4.63b
27.07
8.67a
9.21
5.90a
117.72
8.17
4.05b
9.48
12.73c
7.73
2.02
3.54b
8.63
2.70
20.19b
79.24b

22.06
3.19
0.70
25.75
5.27a
26.75
5.66b
7.95
4.31b
107.59
8.95
5.09a
10.65
16.49ab
7.49
2.01
4.03ab
7.79
2.05
23.52ab
88.89ab

21.34
3.26
0.80
26.63
4.72ab
25.70
6.11b
8.43
4.98ab
108.05
0.66
0.40
0.86
1.25
0.55
0.15
0.19
0.66
0.26
1.68
4.83

1.26
0.29
0.07
1.25
0.22
1.85
0.53
0.53
0.35
4.49
0.41
0.03
0.13
0.01
0.82
0.34
0.01
0.23
0.58
0.03
0.08

0.54
0.22
0.22
0.11
0.01
0.92
0.01
0.35
0.01
0.19
a–cValues within rows with different superscripts differ (P < 0.05).
1SBM = soybean meal; HPDDG = high protein dried distillers grains with solubles; CM = canola meal;
DDGS = dried distillers grains with solubles. Dietary treatments consisted of 4 different protein sources (SBM,
HPDDG, CM, and DDGS) intended to supply equal amounts of protein (approximately 38% of the total di-
etary protein) from the respective supplemental protein sources.

Page 8

Journal of Dairy Science Vol. 93 No. 5, 2010
CHRISTEN ET AL.
2102
Table 8. Amino acid concentrations (μmol/dL) in plasma from the subcutaneous abdominal vein (LSM) in
lactating dairy cows fed treatment diets1
Item
Diet
SEMP-valueSBM HPDDGCM DDGS
Essential AA, μmol/dL
Arg
His
Ile
Leu
Lys
Met
Phe
Thr
Trp
Val
Total essential AA
Nonessential AA, μmol/dL
Ala
Asp
Asn
Glu
Gln
Gly
Pro
Ser
Tyr
Total nonessential AA

5.62
3.57a
7.89a
9.33a
3.27
0.67b
1.70b
5.77
2.69
20.48a
61.02a

17.29
1.95b
0.41b
17.54b
1.89b
24.32
4.39b
5.91
2.38b
81.97b
4.65
4.06a
6.56ab
11.80a
3.22
1.03a
2.44a
6.30
2.61
19.87a
61.54a

19.17
2.63a
0.59a
21.85a
2.11ab
26.87
6.56a
6.40
4.09a
96.30a
5.67
2.74b
4.93b
6.30b
2.96
0.87ab
1.66b
5.66
2.25
14.64b
47.71b

17.44
1.96b
0.43b
19.39ab
2.35a
25.57
3.87b
4.89
2.26b
83.13b
4.85
3.77a
6.39ab
9.87a
2.78
0.76ab
1.85b
5.66
2.93
17.86ab
56.48ab

17.49
1.93b
0.50ab
18.40ab
2.06ab
25.07
4.50b
5.41
2.78b
85.51ab
0.65
0.38
0.84
1.13
0.39
0.11
0.16
0.62
0.24
1.60
4.07

1.28
0.22
0.06
1.28
0.13
1.82
0.44
0.53
0.32
4.13
0.52
0.01
0.08
0.01
0.69
0.07
0.01
0.79
0.34
0.01
0.02

0.55
0.04
0.05
0.04
0.06
0.69
0.01
0.12
0.01
0.06
a,bValues within rows with different superscripts differ (P < 0.05).
1SBM = soybean meal; HPDDG = high protein dried distillers grains with solubles; CM = canola meal;
DDGS = dried distillers grains with solubles. Dietary treatments consisted of 4 different protein sources (SBM,
HPDDG, CM, and DDGS) intended to supply equal amounts of protein (approximately 38% of the total di-
etary protein) from the respective supplemental protein sources.
Table 9. Arteriovenous differences in AA plasma (LSM) from lactating dairy cows fed treatment diets1
Item
Diet
SEMP-valueSBMHPDDG CMDDGS
Essential AA, μmol/dL
Arg
His
Ile
Leu
Lys
Met
Phe
Thr
Trp
Val
Total essential AA
Nonessential AA, μmol/dL
Ala
Asp
Asn
Glu
Gln
Gly
Pro
Ser
Tyr
Total nonessential AA

3.14
1.29
3.81
6.02
4.71
1.18
1.95
2.50ab
0.39
5.18
29.99

3.89
1.24
0.32
6.71
2.44
2.61a
1.87
2.65
1.88
23.85
2.89
1.25
4.16
6.86
4.29
1.14
1.96
3.04a
0.76
5.48
31.63

3.90
1.24
0.25
5.74
2.46
1.13b
2.12
2.83
1.81
21.07
2.70
1.31
4.80
6.38
4.77
1.17
1.84
3.05a
0.50
5.50
31.93

3.58
1.22
0.29
6.25
2.93
1.47b
1.79
2.09
2.06
23.83
3.90
1.32
4.27
6.62
4.73
1.25
2.19
2.32b
0.32
5.65
32.43

3.78
1.34
0.31
7.33
2.68
1.08b
1.62
3.09
2.20
23.00
0.56
0.09
0.52
0.32
0.24
0.10
0.14
0.26
0.16
0.32
1.67

0.58
0.13
0.05
0.60
0.16
0.42
0.25
0.36
0.15
1.52
0.42
0.92
0.59
0.27
0.24
0.87
0.29
0.09
0.22
0.74
0.72

0.53
0.89
0.83
0.13
0.11
0.06
0.51
0.72
0.23
0.40
a,bValues within rows with different superscripts differ (P < 0.05).
1SBM = soybean meal; HPDDG = high protein dried distillers grains with solubles; CM = canola meal;
DDGS = dried distillers grains with solubles. Dietary treatments consisted of 4 different protein sources (SBM,
HPDDG, CM, and DDGS) intended to supply equal amounts of protein (approximately 38% of the total di-
etary protein) from the respective supplemental protein sources.

Page 9

study, extraction efficiency (Table 10) indicated that
Met was the first limiting AA by the mammary gland
with SBM, whereas Lys was first limiting with the
other diets. Phenylalanine was third limiting with all
diets but was less limiting (P < 0.05) with the HPDDG
diet than with the other diets. Likewise, Leu, Trp, and
Val were less limiting with the HPDDG diet. If Tyr
were considered to be an essential AA, as it is in most
animal tissues (Schingoethe, 1996), it was less limiting
with the HPDDG diet than with the other diets. Thus,
whereas the first limiting AA were essentially of the
same degree of limitation with all protein supplements
evaluated, there were some indications of improved AA
balance when fed HPDDG.
CONCLUSIONS
The recently available HPDDG is a high protein,
lower fat product of ethanol processing available to the
feed industry. When fed to lactating cows as the source
of supplemental protein, HPDDG was equal to SBM,
CM, and DDGS. Cows fed the HPDDG diet produced
the most casein in the milk protein and thus may have
provided the most desirable AA profile for casein pro-
duction. Extraction efficiency of AA from blood by the
mammary gland indicated that Lys was the first limit-
ing AA for HPDDG, CM, and DDGS diets, whereas
Met was first limiting for the SBM diet.
ACKNOWLEDGMENTS
The authors gratefully acknowledge the farm crew
at the South Dakota State University Dairy Research
Unit (Brookings) for care of the cows and assistance
with data collection. The authors also express appre-
ciation for partial funding by Dakota Gold Research
Association (Sioux Falls, SD) and to Poet Nutrition
(Sioux Falls, SD) for supplying the HPDDG and DDGS
used in this research.
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LACTATION RESPONSE TO 4 PROTEIN SUPPLEMENTS
Journal of Dairy Science Vol. 93 No. 5, 2010
Table 10. Extraction efficiencies1 (%) of essential AA (EAA) from blood to mammary gland (LSM) for
lactating dairy cows fed treatment diets2,3
EAA
Diet
SEMP-valueSBMHPDDG CM DDGS
Arg
His
Ile
Leu
Lys
Met
Phe
Thr
Trp
Val
Tyr
Total EAA
36.3 (6)
27.2b (9)
35.6 (7)
41.6b (5)
60.1 (2)
64.1 (1)
54.0a (3)
31.1 (8)
45.7a (4)
21.4b (10)
44.5a [5]
33.6b
39.2 (4)
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58.8 (1)
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44.2b (3)
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34.5b
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37.4a (7)
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51.1a (4)
63.3 (1)
58.5 (2)
52.2a (3)
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39.9a
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63.2 (2)
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30.6 (8)
45.4a (4)
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44.2a [5]
37.0ab
5.25
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3.54
3.08
2.81
3.99
3.04
3.17
3.26
2.00
3.10
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0.52
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0.17
0.01
0.49
0.16
0.03
0.51
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0.03
0.04
0.04
a,bValues within rows with different superscripts differ (P < 0.05).
1Extraction efficiencies = arteriovenous difference of AA (g/L) × 100/concentration (g/L) of AA in plasma of
coccygeal artery.
2SBM = soybean meal; HPDDG = high protein dried distillers grains with solubles; CM = canola meal;
DDGS = dried distillers grains with solubles. Dietary treatments consisted of 4 different protein sources (SBM,
HPDDG, CM, and DDGS) intended to supply equal amounts of protein (approximately 38% of the total di-
etary protein) from the respective supplemental protein sources.
3Numbers in parentheses indicate the apparent order of limiting AA. Numbers in brackets are for Tyr if it were
considered an essential AA.

Page 10

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