Nonstructural Carbohydrate and Protein Effects on Rumen
Fermentation, Nutrient Flow, and Performance of Dairy Cows
ABSTRACT
Four multiparous Holstein cows fitted
with rumen and duodenal cannulas were
used in a 4 x 4 Latin square with
20-d periods. Four diets were formulated
for high and low rumen availabilities of
nonstructural carbohydrate and protein.
Cows were milked and fed three times
daily. Milk production averaged 39 kg/d
and was unaffected by treatment. Dry
matter intake and 4% FCM production
were increased by 1 kg/d for cows fed
the low rumen-available nonstructural
carbohydrate diets. Milk protein percent-
age was elevated when either the high
rumen-available nonstructural carbohy-
drate or high rumen-available protein
diets were fed. Low rumen-available pro-
tein diets increased duodenal passage of
total essential AA but did not increase
passage of Arg, ne, and Met. Passage of
bacterial N was highest (262 g/d) when
high rumen-available nonstructural car-
bohydrate was combined with high
rumen-available protein and was lowest
when high rumen-available nonstructural
carbohydrate was combined with low
rumen-available protein (214 g/d). Diets
with low rumen availabilities of protein
were not advantageous, possibly because
duodenal passage of one or more of the
limiting AA was not increased. Passage
of bacterial N to the duodenum was
highest when rumen availabilities of both
Received July 9, 1992.
Accepted November 9, 1992.
turrent address: Carl S. Akey Inc., C.S. 5002, Lewis-
burg, OH.
2To whom correspondence should be addressed.
1993 J Dairy Sci 76:1091-1105
J. M. ALDRICH,1 L. D. MULLER,? and G. A. VARGA
Department of Dairy and Animal Science
L. C. GRIEL, JR.
Department of Veterinary Science
The Pennsylvania State University
University Park 16802
nonstructural carbohydrate and protein
were high.
(Key words: rumen availability, non-
structural carbohydrate, protein, rumen
fermentation)
Abbreviation key: DIP = degradable intake
protein, EC = ear corn, HMSC = high mois-
ture shelled corn, NSC = nonstructural carbo-
hydrate, RANSC = rumen-available NSC,
RAP = rumen-available protein, UIP = un-
degradable intake protein.
INTRODUCTION
Supplementation of diets for dairy cows for
higher amounts of protein to the small intestine
is justified because high producing cows re-
quire more protein than can be provided by
rumen microbes (17). However, higher percen-
tages of rumen-undegradable intake protein
(UIP) that are fed to cows to increase the
amount of total protein flowing to the small
intestine has not always increased milk
production (37). Two plausible explanations
for a lack of response to increased UIP are that
1) microbial protein synthesis is reduced be-
cause substitution of UIP sources for degrada-
ble protein (DIP) sources deprives rumen mi-
crobes of adequate available N, and,
consequently, the supply of protein to the cow
may not be changed, or 2) higher UIP fails to
increase the intestinal supply of limiting AA.
Optimization of microbial growth may be a
more logical strategy than diet formulation
with higher percentages of UIP because rumen
microbes supply high quality protein to the
cow (3) and because end products of rumen
fermentation supply most of the host’s energy
requirement (33).
Microbial growth depends on a source of
rumen-available carbohydrate to supply ATP
1091
1092 ALDRICH ET AL

for biosynthesis of cell material (22). To
achieve optimal growth, the rate of ATP
production (carbohydrate fermentation) must
equal its utilization (protein synthesis), or fer-
mentation becomes uncoupled (12). This need
for balance implies that rumen availabilities of
carbohydrates and protein should be matched
to maximize microbial protein synthesis. This
hypothesis is supported by the work of
Herrera-Saldana et al. ( l l ) , who showed that
microbial protein synthesis was maximized
when starch and protein sources (barley and
cottonseed meal), which have relatively rapid
rates of digestion, were fed. Exact amounts of
rumen-available carbohydrate and protein that
maximize microbial yields are not known.
Hoover and Stokes (14) suggested that, when
high percentages of grain are fed to high
producing dairy cows, DIP may be more limit-
ing than carbohydrate for microbial growth.
The objective of this experiment was to
evaluate the effects of diets that vary in rumen-
available nonstructural carbohydrate (RANSC)
and protein (RAP)on bacterial protein synthe-
sis, rumen fermentation, flow of nutrients to
the small intestine, milk production, and milk
component yield.
MATERIALS AND METHODS
Four multiparous Holstein cows (50 rt 8
DIM at the onset of the trial) were used. Cows
were fitted with rumen and duodenal cannulas.
The latter was a Y-type plastisol cannula with
2.5 cm i.d. and 135°C internal flange angle
(provided by B. P. Glenn, USDA-ARS, Belts-
ville, MD). The cannula was placed approxi-
mately 15 cm anterior to the bile ampulla.
Cows were housed in individual tie stalls and
milked and fed three times daily at 0600, 1400,
and 2200 h.
The experimental design was a 4 x 4 Latin
square with 20-d periods. The four treatments
consisted of diets formulated to contain high
and low RANSC and high and low RAP. The
treatments were 1) high RANSC and high
RAP, 2) high RANSC and low RAP, 3) low
RANSC and high RAP, and 4) low RANSC
and low RAP. Ingredient composition of the
four diets is listed in Table 1. For high and low
RANSC, high moisture shelled corn (HMSC)
and dry ear corn (EC) were used, respectively;
for high and low RAP, canola meal and blood
meal were used, respectively. The EC was
ground using a 2.2-cm screen. The HMSC was
stored in a sealed silo, obtained every 4 to 5 d
from a nearby commercial dairy farm, rolled,
and stored in plastic containers at 4°C. Soy-
bean meal (48% CP) was added to all diets as
a source of RAP so that low RAP diets were
not grossly deficient in an available N source
for rumen microbes (26). High RANSC diets
contained more corn silage and more total

TABLE 1. Ingredient composition of experimental diets formulated for high and low xumen-available nonstructural
carbohydrate (RANSC) and high and low rumen-available protein (RAP) as a percentage of the DM.
High RANSC Low RANSC
High LOW High Low
Ingredient RAP RAP RAP RAP
Corn silage 23.5 37.1 14.0 24.3
Alfalfa-grass silage 25.2 16.0 26.0 20.7
High moisture shelled corn 31.7 30.7 . . . ...
Ear corn ... ... 39.5 38.8
Canola meal 7.0 ... 7.8 ...
Blood meal ... 4.8 . . . 4.3
soybean meal 7.3 5.7 7.4 6.2
Molasses 3.0 3.0 3.0 3.0
Urea .1 .1 .1 .1
Mineral-vitamin premix' 2.2 2.6 2.2 2.6

'High RAP contained dicalcium phosphate, 27.2%; sodium bicarbonate, 21.9%; trace-mineralized salt, 21.9%;
limestone, 11.4%; CaSO4, 4.4%; Mg03, 4.4%; Se premix (200 ppm), 5.7%; vitamin A, 650,400 IUkg; vitamin D,
453.600 IUkg; vitamin E, 4760 I U k g , 3.6%. Low RAP contained dicalcium phosphate, 34.6%; sodium bicarbonate,
19.3%; trace-mineralized salt, 19.3%; limestone, 11.6%; CaS04, 3.9%; MgOg, 3.9%; Se premix (200 ppm), 4.6%;
vitamin A, 650,400 IUlkg; vitamin D. 453,600 IUkg; vitamin E, 4760 IUkg, 3.0%.

Journal of Dairy Science Vol. 76, No. 4, 1993

 NONSTRUCTURAL CARBOHYDRATE AND PROTEIN
forage than low RANSC diets (to maintain
similar NEL concentrations), and high RAP
diets contained more alfalfa-grass silage than
low RAP diets.
The first 3 d of each period were used to
adjust cows gradually to their new diets; 25,
50, and 75% of the current period's diet and
75, 50, and 25% of the previous period's diet
were offered on d 1, 2, and 3, respectively. On
d 4, cows received 100% of their experimental
ration. Diets were prepared by weighmg each
ingredient and blending in a drum-type mixer
(Data Ranger@';American Calan, Inc., North-
wood, NH). The amount offered was adjusted
daily to achieve 5 to 10% orts. The percen-
tages of the total daily allotment fed were 38,
35, and 27% at the morning, afternoon, and
evening feedings, respectively.
Diets were formulated to provide nutrients
for production of 40 kg of milk (21). Silages
were sampled once weekly for the first 2 wk of
each period and analyzed for CP. The amount
of corn (HMSC or EC) and soybean meal were
adjusted to maintain a ration CP of 17.5%
when the calculated CP in the ration was less
than 17.3% or greater than 17.8%. Amounts of
corn and soybean meal (Table 1) are the aver-
age amounts fed throughout the experiment. In
no case did a ration adjustment in the amount
of corn or soybean meal fed change by more
than .2 kg of DM.
Samples of the TMR and orts were obtained
daily during d 14 to 20 of each period, frozen
(-20°C) immediately, composited by cow,
dried at 55°C for 48 h, ground through a Wiley
mill (1-mm screen; Arthur H. Thomas,
Philadelphia, PA), and stored for subsequent
analyses. Dry matter intake was calculated for
d 17 to 20 of each period to correspond to
duodenal and fecal samplings.
At each milking during d 14 to 20 of each
period, milk weights were recorded, and sam-
ples were then taken. Samples were preserved
with 2-bromo-2-nitropropane-1,3 diol and ana-
lyzed for fat, protein, and SCC at the Pennsyl-
vania DHIA milk testing laboratory (Foss
605B Milko-Scan; Foss Electric, Hiller@d,
Denmark).
Duodenal digesta (approximately 500 ml)
were collected every 4 h on d 18 to 20 of each
period; collection time advanced 1 h each 24 h
for a total of 18 samples. Stainless steel diver-
sion tubes were inserted into the cannula dur-
1093
ing sampling to ensure that flow originated
from the abomasum. The pH was measured
immediately by glass electrode, and 200 ml of
digesta were added to a common container for
each cow and frozen (-2O'C). Samples were
thawed at room temperature (2133, and an
aliquot was centrifuged (1000 x g for 10 min
at 4°C) and analyzed for N H 3 N (5). The
remaining portion was lyophilized and ground
through a Wiley mill (1-mm screen).
Rumen samples were obtained on d 18 to
20 of each period at 0, 1.5, 3, 6, and 8 h after
the morning feeding. Grab samples were ob-
tained from the midventral, anterior, and dorsal
regions. In addition, 750 ml of liquid from the
ventral region were obtained and added to the
grab samples for a total volume of approxi-
mately 4 L. This sample was mixed by hand,
and a I-L subsample was processed through a
food processor (Kenmore@' model number
116965; Sears, Roebuck and Co., Chicago, IL)
using a chopper blade at the fastest speed.
Approximately 3 min were needed to load the
processor, which then was allowed to run for
an additional 30 to 60 s. Contents from the
processor were squeezed through two layers of
cheesecloth. The pH was measured immedi-
ately by glass electrode. A 250-ml aliquot was
preserved with 21 ml of 37% formaldehyde in
.9% saline solution and frozen (-20°C) for
subsequent isolation of bacteria. Another por-
tion (5 ml) was extracted, 1 ml of 25%
metaphosphoric acid and 1 ml of 2-ethyl bu-
tyric acid (internal standard) were added, and
the sample was frozen immediately (-20°C).
For analysis, the latter samples were thawed at
room temperature (2 1°C) and centrifuged
(lo00 x g for 20 min at 4"C), and a portion of
the supernatant was analyzed for NH3 N. A
second portion was centrifuged (3000 x g for
20 min at 4'C), and the supernatant was ana-
lyzed for VFA (8). For bacterial isolation,
samples were thawed slowly at 4°C and cen-
trifuged (lo00 x g for 10 rnin at 4°C) to
remove protozoa and feed particles; the super-
natant was drawn off and centrifuged (18,400
x g for 30 min at 4 T ) , washed with 25 ml of
3 5 % saline, and recentrifuged (18,400 x g for
30 min at 4°C). The supernatant was discarded,
and the bacterial pellet was washed twice with
25 ml of deionized water and frozen at -20°C.
Bacterial isolates then were lyophilized and
analyzed for DM, ash, N (l), and purines (38).
Journal of Dairy Science Vol. 76, No. 4, 1993
1094 ALDRICH ET AL
The N:purine ratio was used to calculate the
flow of bacterial N to the small intestine.
Fecal grab samples were obtained at every
second duodenal sampling time (9 samples per
period), frozen (-20°C) immediately, thawed at
the end of the period, composited by cow,
dried at 55°C and ground through a Wiley
mill (1-mm screen).
Blood was drawn (20 ml) from the coc-
cygeal vein at 0900 and 1700 h on d 19 and 20
of each period into heparinized tubes and cen-
trifuged (3000 x g for 15 min at 4'C), and the
plasma was frozen (-2O'C) for analysis of urea
N (Technicon Autoanalyzer@; Technicon
Corp., Tarrytown, NY).
Duodenal digesta flow and digestibilities
were determined using Yb-marked insoluble
DM. Ytterbium was bound to the insoluble
DM of each of the four TMR. Each diet was
soaked in tap water for 24 h with occasional
stirring. Soaked material was rinsed with tap
water, filtered through three layers of cheese-
cloth, and hand squeezed. Insoluble material
then was soaked in tap water in a ratio of 1:3
(wt/wt), and .6 g of YbCL3.6H20 were added/
100 g of wet material. After 48 h with occa-
sional stirring, the Yb-marked feed was
washed and filtered as previously described
and dried at 60'C for 48 h. After air equilibra-
tion, 120 g of Yb-marked insoluble DM were
weighed into 1-L plastic bags for dosing.
Marker was dosed into the rumen at 0600 and
1630 h on d 11 to 20 of each period. Measured
Yb concentration averaged 14.1 mg/g of in-
soluble DM across the four diets.
Diets, duodenal digesta, and feces were ana-
lyzed for DM, CP, and ash (l), ADF, NDF,
ADF CP, and NDF CP (25), and Yb (10).
Organic matter truly digested in the rumen was
calculated as OM intake minus the difference
between OM and bacterial OM flow to the
duodenum. An equivalent calculation was
made for N.
Protein solubility of diets was determined
using a borate-phosphate buffer (15). For anal-
ysis of nonstructural carbohydrate (NSC),sam-
ples were ground using a UDY@ cyclone mill
(1-mm screen; UDY Corp., Ft Collins, CO);
the procedure of Smith (30), modified to use
ferricyanide as the colorimetric indicator, was
used. Amino acid analysis was by the hydroly-
sis procedure described by Lynch et al. (18),
except that the hydrolysate was passed through
a Whatman filter (Number 541; Whatman In-
ternational, Ltd., Maidstone, England).
Journal of Dairy Science Vol. 76, No. 4, 1993
The four diets were evaluated in situ to
determine RANSC and RAP. Fresh samples
were ground with dry ice using a Wiley mill
(4-mm screen). An amount to yield 5 g of DM
was weighed into duplicate 10- x 20-cm poly-
ester bags (Ankom, Fairport, NY).Bags had a
mean pore size of 53 f 10 p , and sample size
(DM) to surface area ratio was 12.5 mg/cm2.
Bags were incubated in the rumen of 2 cows
per period (in situ diet was the same as the diet
fed) beginning on d 19 for 0, 1, 2, 3, 6, 12, 18,
24, 48, and 72 h after soaking at 39°C in
distilled water for 15 min. Two blank bags
were included in each incubation and used to
correct for material entering the bags. Bags
were inserted in reverse order and retrieved as
a group, rinsed in cold tap water, and washed
in a washing machine under the gentle cycle
for 10 min. Bags then were dried at 55°C for
48 h. The residues at 0 to 24 h were analyzed
for NSC and N as described previously. A
linear regression model was fitted to the log-
transformed residuals to determine disappear-
ance rates. Rumen availability was calculated
as
RANSC or RAP = a + {b X [kd/(k,j + kp)l)
where
a = 100 - antilog intercept (amount
soluble at 0 h),
b = 100-a,
k,j = digestion rate of b, and
k, = passage rate (assumed to be .07/h).
For the 16 regressions, r2 2 .93 except for one
replicate for CP disappearance (high RANSC
and low RAP; r2 = .86).
Data (except in situ data) were analyzed as
a 4 x 4 Latin square using the general linear
models procedure of SAS (27); cow, period,
and treatment were the effects in the model.
Single degree of freedom orthogonal contrasts
were used to determine the effect of RANSC,
RAP, and their interaction. Significance was
declared at P < .lo.
RESULTS AND DISCUSSION
In Situ Evaluation and Diet Composition
The in situ estimates of RANSC (as a per-
centage of NSC) was 17% higher for the high
 NONSTRUCTURAL CARBOHYDRATE AND PROTEIN 1095
TABLE 2. In situ estimates of rumen availability' of nonstructural carbohydrate (NSC) and protein for experimental diets
formulated for high and low rumen-available NSC (RANSC) and high and low rumen-available protein (RAP).
High RANSC Low RANSC
High Low High Low
Item RAP RAP RAP RAP SEM
RANSC, 9% of NSC 80.7 82.1 71.1 67.6 2.9
RAP, % of CP 65.7 52.4 65.2 51.1 3.6
RANSC:RAP 2.5 3.4 2.2 2.6 .I
lRumen availability calculated as RA = a + {b X kd(kd + k )]]where a = 100 - antilog of the intercept (soluble and
53 I(filterable); b = 100 - a; = digestion rate of b; a n 8 kp = passage rate (.07/h).

RANSC than for the low RANSC diets (81.4

vs. 69.4%), and RAP (as a percentage of pro-
tein) was 26% higher for the high RAP diets
than for the low RAP diets (65.5 vs. 51.8%;
Table 2). The ratio of RANSC:RAP was 2.5
and 2.6 for the high RANSC and high RAP
diet and the low RANSC and low RAP diet,
respectively. The high RANSC and low RAP
diet had the highest ratio (3.4), and the low
RANSC and high RAP diet had the lowest
0123 6 12 18 24 ratio (2.2). The amount of NSC solubilized at
0 h was considerably higher for the high
RANSC diets. Disappearance rates of NSC
were higher for the high RAP diets than for the
80
low RAP diets (Figure lA), but the reason for
B this difference is unknown. For N digestion,
both rates and rumen availabilities were hgher
for the high RAP diets than for the low RAP
diets (Figure 1B).
The suggested percentage of UIP in diets
for high producing cows is 6.0 to 6.3% of
ration DM (21). These diets averaged 6.1 and
8.5% UIP for the high and low RAP diets,
respectively, when UIP was calculated from
0' I
0123 6 12 18 24 RAP measured in situ (Table 3). Diets initially
Hour
were formulated (for the high and low RAP
diets, respectively) to be less than or greater
Figure 1. In situ disappearance of A) nonstructural than NRC (21) recommendations for UIP. All
carbohydrate (NSC) and B) N for diets containing high three protein sources used in the diets had
mrnen-available NSC (RANSC) and high rumen-available
protein (RAP) (7).high RANSC and low RAP (V), low higher in situ UIP (blood meal, 90.4%; canola
RANSC and high RAP (0).and low RANSC and low RAP meal, 41.6%; and soybean meal, 32.4%; in situ
(0). Rates of disappearance (percentage per hour) of NSC data for individual ingredients are not shown)
and N were 11.9 and 5.2 for high RANSC and high RAP, than estimated when the diets originally were
8.0 and 1.8 for high RANSC and low RAP, 12.8 and 6.1
for low RANSC and high RAP, and 8.0 and 1.8 for low formulated. Therefore, UIP for all diets was
RANSC and low RAP, respectively. slightly higher than expected.
Journal of Dairy Science Vol. 76, No. 4, 1993
I096 ALDRICH ET AL

TABLE 3. Chemical composition of experimental diets formulated for high and low rumen-available nonstructud
carbohydrate (RANSC) and high and low rumen-available protein (RAP).
High RANSC Low RANSC
High Low High Low
Item RAP RAP RAP RAP SEM'
DM, % 60.6 53.7 68.2 61 .O 1.4
NEL.' McaVkg of DM 1.64 1.66 1.61 1.61 . .
(70of DM)
CP 17.5 17.3 17.6 17.8 .I
Soluble CP 5.9 5.4 5.4 5.1 .2
UIP~ 6.0 8.2 6.1 8.7 . . .
ADF 21.9 20.1 21.2 21.1 .3
NDF 34.2 34.4 35.6 35.6 .4
Forage NDF 25.1 26.5 21.0 23.0 . . .
NSC4 35.8 37.7 35.0 34.6 .5
Ash 6.2 5.3 6.0 5.6 .1
ADF-Insoluble CP 1.1 .9 1.2 1 .o <.1
NDF-Insoluble CP 2.9 3.6 2.9 3.7 .1
IStandard error of the pooled treatment means (n = 16).
'Estimated from NRC (21).
3Calculated as ( 1 0 0 - rumen-available protein)/100 x CP. Rumen-available protein measured in situ.
4Nonstructural carbohydrate

The NSC content was 2 percentage units
higher for the high RANSC (36.8%) than for
the low RANSC (34.8%) diets (Table 3). Acid
detergent-insoluble CP was low for all diets,
and NDF-insoluble CP was higher for the low
RAP diets than for the high RAP diets, most
likely because of the blood meal in the low
RAP diets. Blood meal contained 37% NDF-
insoluble CP.
Processing corn resulted in relatively small
particle sizes; 67 and 88% of the particles were
less than or equal to 3.35 mm for HMSC and
EC, respectively (Table 4). Because processing
can affect rumen digestion rate and availability
of corn (9).the objective was to minimize the
differences in particle size of the two forms of
corn; however, the rolled HMSC had a higher
proportion of larger particles (66 vs. 48%
21.17 mm) than did the ground EC.
of OM digested in the rumen was similar
across diets; however, the higher flow of OM
to the small intestine for cows fed the low
RANSC diets did not result in compensatory
digestion of OM in the lower gastrointestinal
tract because the quantity of OM apparently
digested in the total tract was higher for cows
fed the high RANSC diets (15.9 vs. 15.1 kg/d;
P < .01). Postruminal OM digestion tended ( P
e .15) to be higher (3.9 vs. 3.3 kg/d) for cows
fed the low RAP diets, apparently because of
TABLE 4. Particle size distribution of rolled high moisture
shelled corn (HMSC) and ground ear corn (EC).
Screen
size HMSC EC SEI
- (46 retained on screen)' -
~

Intake and Duodenal flow of OM, 13.35 66.5 87.8** 2.6

NSC, and Fiber Components 22.38 45.6 21.6** 2.3
21.17 65.5 47.8** .6
Cows fed the low RANSC diets consumed <.43 15.0 8.1 3.8
<.I 1 3.7 .9 1.6
slightly more DM and OM (1 kg/d; P c .OS)
than those fed the high RANSC diets (Table 'Standard error of difference (n = 2).
5 ) , and OM passage to the small intestine was T o m was oven-dried at 60'C for 24 h after process-
1.4 kg/d higher (12.8 vs. 11.4 kg) when cows ing.
were fed the low RANSC diets. The proportion **Differs from HMSC (P < .01).

Journal of Dairy Science Val. 76, No. 4. 1993

 NONSTRUCTURAL CARBOHYDRATE AND PROTEIN 1097
TABLE 5. Effect of amount of rumen-available nonstructud carbohydrate (RANSC) and rumen-availableprotein (RAP)
on DMI and duodenal passage, and digestibilities of OM and nonstructud carbohydrate (NSC).
Hieh
" RANSC Low R A N K
Effect (P <)
High Low High Low
Item RAP RAP RAP RAP SEM Cl P2 C x P3
BW, kg 710 717 710 712 5 NS4 NS NS
DMI, kg/d 25.0 24.9 26.7 25.3 .5 .08 NS NS
OM
Intake. kg/d 23.4 23.5 25.1 23.9 .5 .08 NS NS
Passage to duodenum, kg/d 11.6 11.2 12.6 13.0 .7 .09 NS NS
Digested in rumen apparently, % 50.6 52.3 49.8 45.8 2.1 .13 NS NS
Digested in rumen truly, % 62.7 62.5 61.3 56.9 3.3 NS NS NS
Digested postruminally, kg/d 3.7 3.8 2.8 4.1 .4 NS .I5 NS
Total tract digestion, kg/d 15.5 16.2 15.3 14.9 .3 .07 NS NS
% of OM intake 66.4 68.7 61.1 63.0 .8 .01 NS NS
NSC
Intake, kg/d 8.9 9.4 9.3 8.8 .3 NS NS NS
Passage to duodenum, kg/d 1.8 2.1 2.6 3.1 .4 .04 NS NS
Digested in rumen apparently, % 80.4 78.3 71.9 65.3 3.3 .02 NS NS
Total tract digestion, kg/d 8.3 8.7 8.3 7.9 .2 NS NS .IO
% of NSC intake 92.6 92.6 89.5 89.8 .6 .01 NS NS
IC = The RANSC effect.
2P = The RAP effect.
3C x P = The interaction between RANSC and RAP.
4P > .10

the higher amount of feed protein escaping
rumen fermentation in these diets (Table 8).
The percentages of NSC digested in the
rumen (P < .02) and the percentages dqested
in the total tract (P < .01) were higher for cows
fed the high RANSC diets than for those fed
the low RANSC diets (Table 5). The amount
of NSC digested postruminally was higher for
cows fed the low RANSC diets than for those
fed the high RANSC diets (1.9 vs. 1.3 kg/d; P
e .07). The higher total tract apparent digesti-
bility of NSC for the high RANSC diets com-
pared with the low RANSC diets (92.6 vs.
89.7%, respectively) was due to the higher
rumen digestibility of the high RANSC diets.
A RANSC by RAP interaction existed on the
quantity of NSC digested in the total tract.
This interaction may have been caused by
slight but nonsignificant (P > .lo) differences
in NSC intake among diets.
The intake of NDF was higher (P < .01) for
the low RANSC diets than for high RANSC
diets (9.2 vs. 8.5 kg/d; Table 6), which
reflected higher NDF content of the low
RANSC diets and higher DMI (Table 5 ) .
Digestion of NDF (50.6 vs. 55.2%) and ADF
(49.1 vs. 55.4%) in the rumen were affected
adversely by the high RAP diets compared
with the low RAP diets (P e .08). The high
RAP diets appeared to promote a more vigor-
ous fermentation, as evidenced by the higher
total VFA concentration (Table 7). Degrada-
tion of NSC proceeded at a faster rate (Figure
1A) for the high RAP dets than for the low
RAP diets (12.3 vs. 8.0%/h). Rumen pH also
was lower for the high RAP diets at 6 and 8 h
postfeeding (Figure 2), which would be ex-
pected to correspond to the time of maximal
fiber digestion (16). The higher rate of NSC
digestion with the high RAP diets could have
reduced fiber digestion because of its effect on
rumen pH or because of competition for sub-
strate between fiber- and NSC-digesting organ-
isms (13). Differences in fiber digestibility be-
tween the high and low RAP diets also may
have been the result of differences in the rela-
tive proportions of corn and alfalfa-grass
silages in these diets (Table 1). Alfalfa con-
tains a higher amount of lignin and has a faster
rate, but lower extent, of digestion than do
grasses (31).
Intake sometimes is reduced when a more
readily fermentable NSC source is substituted
Journal of Dairy Science Vol. 76, No. 4, 1993
1098 ALDRICH ET AL
TABLE 6. Effect of amount of rumen-available nonstructural carbohydrate (RANSC) and rumen-available protein (RAP)
on intake, duodenal passage, and digestibilities of ADF and NDF.
High RANSC Low RANSC
Effect ( P <)
High Low High Low
Item RAP RAP RAP RAP SEM C1 P2 c x P3
ADF
Intake, kg/d 5.5 5.0 5.7 5.3 .2 NS4 .07 NS
Passage to duodenum, kg/d 2.8 2.2 2.8 2.4 .2 NS .02 NS
Digested in rumen apparently, 96 48.5 56.2 49.7 54.6 3.0 NS .08 NS
Digested postruminally, kg/d -.5 -.5 -.9 -.8 .I .05 NS NS
Total tract digestion, kg/d 2.1 2.3 2.0 2.1 .2 NS NS NS
% of ADF intake 39.4 45.5 35.6 39.4 2.1 .06 .06 NS
NDF
Intake, kg/d 8.5 8.6 9.5 9.0 .2 .01 NS NS
Passage to duodenum. kg/d 4.3 3.7 4.5 4.1 .2 NS .08 NS
Digested in rumen apparently, t 48.8 55.8 52.4 54.6 1.7 NS .04 NS
Digested postruminally. kg/d -.7 -.7 -1.2 -1.0 .2 .ll NS NS
Total tract digestion, kg/d 3.5 4.1 3.8 3.9 <.1 NS .01 .05
% of NDF intake 41.5 47.3 40.7 44.4 .8 .06 .01 NS
'C = The RANSC effect.
zP = The RAP effect.
3C x P = The interaction between RANSC and RAP.
4P .2 .lo.

for a less fermentable source (4, 20). Whether
this reduction is because of a lower rumen pH
(20) or because of the palatability of the NSC
source (barley) is uncertain, although the
reduction in intake on barley diets observed by
Casper et al. (4) was not associated with a
reduction in rumen pH. In contrast, Herrera-
Saldana et al. (11) observed no differences in
intake when barley replaced milo in diets for
lactating cows.
Rumen pH, NH3 N, VFA,
and Plasma Urea N
Rumen pH was significantly lower (6.28 vs.
6.39) when cows were fed the high RAP diets
( P < .01, Table 7). Cows fed the low RANSC
diets also had lower (P < .07) rumen pH, but
the magnitude of the difference was small. The
change in pH over time postfeeding (Figure 2)
indicates that the low RANSC diets had
smaller fluctuations than the high RANSC
diets, which is particularly evident between 1.5
and 6 h postfeeding. The difference in pH at
these hours may have been the result of the
higher fraction of soluble NSC (Figure 1A) for
the high RANSC diets than for the low
Journal of Dairy Science Vol. 76, No. 4, 1993
RANSC diets. This fraction is assumed to be
degraded rapidly in the rumen. At 6 and 8 h
postfeeding, rumen pH was affected more by
protein availability when the high RAP diets
produced lower rumen pH.
The high RAP diets also stimulated higher
total VFA (P < .08) concentrations in the
rumen (145.8 vs. 137.1 pmoVml; Table 7). In
continuous culture fermenters, total VFA
production was increased linearly as DIP
(peanut meal) increased (32). Although
RANSC had no effect on acetate concentra-
tions, propionate concentrations tended to be
higher (30.7 vs. 27.0 pmoVml; P = .16), and
acetate to propionate ratios were lower (3.08
vs. 3.49), for cows fed the high RANSC diets
(P < .03). Cows fed the high RAP diets also
had lower (3.12 vs. 3.45; P < .07) ratios
because of higher concentrations of propionate
(31.3 vs. 26.4 pmoVml; P < .07). These ratios
may have resulted from the considerable quan-
tities of propionate derived from the degraded
protein (35). Butyrate concentrations were
lower (16.3 vs. 18.8 pmoVml; P < .06), and
isovalerate concentrations were higher (2.2 vs.
1.9 pmoVm1; P < .04),when cows were fed
 NONSTRUCTURAL CARBOHYDRATE AND PROTEIN 1099
Production of C4 and C5 VFA would be
expected to be higher when more DIP sources
are fed; however, measured concentrations re-
flect both production and utilization by rumen
6.5 - microbes; therefore, isobutyrate and valerate,
but not isovalerate, increased when the high
6.4 -
I RAP diets were fed. Others, also (4, 20) noted
Q
6.3 - variable effects of carbohydrate and protein
6.2 - sources on concentrations of the minor VFA.
When a less fermentable NSC source (corn)
6.1 - was fed, butyrate concentrations were higher
6.0 - than when a more fermentable NSC source

'.' ' 0 1.5 3

Hour
6 8
I (barley) was fed (4), which agrees with results
Figure 2. Rumen pH for cows fed diets containing high
rumen-available NSC (RANSC) and high rumen-available
protein (RAP) (V),high RANSC and low RAP (V). low
RANSC and high RAP (0).and low RANSC and low RAP
(0).
the high RANSC diets, and concentrations of
isobutyrate (1.2 vs. 1.1 pmoVml; P c .06) and
valerate (2.5 vs. 2.0 FmoVml; P e .03) were
higher when cows were fed the high RAP
diets.
of the present study; however, isovalerate con-
centrations were higher when the less ferment-
able NSC source was fed. When corn, barley,
soybean meal, or fish meal was fed, McCarthy
et al. (20) observed no effect on molar propor-
tions of butyrate, valerate, or isovalerate.
Rumen N H 3 N concentrations were lower
(9.6 vs. 13.6 mg/dl) when cows were fed low
RAP diets (P < .01) or high RANSC diets
(10.6 vs. 12.6 mg/dl, P < .07); however, the
pattern of NH3 N concentration across sam-
pling time was similar for all diets (Figure 3).
Russell et al. (26) found that N H 3 accumula-
tion was inversely proportional to the rate of
carbohydrate addition. Mahadevan et al. (19)

TABLE 7. Effect of amount of rumen-available nonstxuctural carbohydrate (RANSC) and rumen-available protein (RAP)
on rumen pH, NH3 N, VFA, and plasma urea N.
High RANSC Low RANSC
Effect (P <)I
High Low High Low
Item RAP RAP RAP RAP SEM C* Pi
PH 6.29 6.40 6.26 6.37 .01 .07 .01
NH3 N, mgldl 12.4 8.9 14.8 10.4 .9 .07 .o1
Plasma urea N, mddl 15.9 17.1 16.8 15.7 .7 NS4 NS
(pmoUml)
Total VFA 147.9 135.6 143.7 138.7 4.1 NS .08
Acetate (A) 90.7 86.5 90.7 90.7 1.9 NS NS
Propionate (P) 33.9 27.4 28.7 25.3 2.2 NS .07
A:P 2.9 3.3 3.3 3.6 .1 .03 .07
Butyrate 17.1 16.4 18.9 17.7 .7 .06 NS
Isobutyrate 1.2 1.1 1.2 1.1 <.1 NS .06
Valerate 2.7 2.0 2.3 2.0 .2 NS .03
Isovalerate 2.3 2.1 1.9 1.9 .1 .04 NS
'The interaction between RANSC and RAP was not significant (P > .lo).
2C = The RASNC effect.
3P = The RAP effect.
4P > .lo.

Journal of Dairy Science Vol. 76, No. 4, 1993

1100 ALDRICH ET AL.
25
20
- be the result of the contribution of endogenous
9
E" l 5
z-
10
z tine. Also, an interaction (P c .03) and a
5 efficiency (expressed either as grams of bac-
0 3
0 1.5 6 0
Hour
Figure 3. Rumen NH3 N concentration for cows fed
diets containing high rumen-available NSC (RANSC) and
high rumen-available protein (RAP) (V),high RANSC
and low RAP ('7). low RANSC and high RAP (e), and
low RANSC and low RAP (0).
suggested that the lower NH3 concentrations
resulting when higher energy diets were fed
were attributed to the increased availability of
soluble carbohydrates, which provided more a-
keto acids for NH3 fixation. Mean NH3 N
concentrations for all treatments were higher
than the 5 mg/dl suggested to be optimal for
maximum microbial protein synthesis (28);
however, concentrations approached this level
at 6 and 8 h postfeeding for the low RAP diets.
Despite the differences in rumen NH3 N con-
centrations across diets, no significant effect of
RANSC or RAP percentages existed on
plasma urea N concentration (Table 7).
Passage of N to the Small Intestine
and Bacterial Protein Synthesis
Intake of N was higher (739 vs. 696 g/d; P
c .04)on the low RANSC diets because of the
higher DMI of these diets; however, passage to
the small intestine was not affected by RANSC
(Table 8). Passage of N ( P c .15) and NAN (P
c .12) to the small intestine tended to be
elevated on the low RAP diets. The low
RANSC and low RAP diet had considerably
higher duodenal N flow than the other three
diets. The proportion of N truly digested in the
rumen was higher (58.1 vs. 45.2%; P < .04)
Journal of Dairy Science Vol. 76, No. 4, 1993
for the high RAP diets. The relative differ-
ences in RAP between diets measured in vivo
were similar to the in situ estimates (Table 2),
but the absolute values were lower, which may
N sources to the quantity of N in the duode-
num. A RANSC by RAP interaction (P c .03)
existed on bacterial N flow to the small intes-
RANSC effect (P c .OS) occurred for bacterial
terial N per kilogram of OM truly digested or
as NSC apparently digested in the rumen,
respectively). Bacterial yield (passage to the
duodenum) and efficiency were highest when
cows were fed the high RANSC and high RAP
diet. Bacterial yield was lower in our study
than that reported in other studies (1 1, 20) with
cows at similar or lower intakes. The N:purine
ratio was used to calculate flow of bacterial N
to the small intestine; these ratios (high
RANSC and high RAP, .77; high RANSC and
low RAP, .80; low RANSC and high RAP,
.79; and low RANSC and low RAP, .86) were
similar for all treatments (P > .lo) and similar
to ratios reported by McCarthy et al. (20)
(average of four treatments = .78). The differ-
ences in bacterial N flow compared with that
found by McCarthy et al. (20) likely were
because of a lower estimate in the present
study of OM flow to the small intestine,
which also tends to lower the estimates for
bacterial efficiency because of higher esti-
mated OM digestion in the rumen.
The effect of RANSC and RAP on passage
of bacterial N agrees with the results of
Herrera-Saldana et al. (1 1). who reported that
passage of microbial protein to the small intes-
tine was highest when starch and protein
degradabilities were synchronized for fast rates
of degradation (barley and cottonseed meal).
In the present trial, soybean and canola
meals were the primary ingredients used to
increase RAP in the high RAP diets, contribut-
ing 46% of the total CP. A major portion of
the N degraded from these two sources is true
protein, which would provide peptides, AA, or
both. Peptides and AA stimulated microbial
growth in vitro (2). Because more NSC was
digested when high the RANSC diets were fed
and because microbial growth is driven
by carbohydrate fermentation (2, 26), high
 NONSTRUCTURAL CARBOHYDRATE AND PROTEIN
RANSC and high RAP may have acted syner-
gistically to enhance bacterial protein synthe-
sis. On the high RANSC and low RAP diet,
the ratio of RANSC:RAP was 3.4 (Table 2);
the higher availability of NSC relative to RAP
could have resulted in uncoupled fermentation
and, hence, reduced bacterial yield and eff-
ciency. Nocek and Russell (22) predicted that a
diet formulated for high rumen availabilities of
both carbohydrate and protein digestion would
maximize microbial protein synthesis com-
pared with diets unmatched in rumen availabil-
ity of carbohydrate and protein or to those low
in both rumen-available carbohydrate and pro-
tein. Results of the present study, which only
considers NSC rather than total carbohydrate,
appear to support their hypothesis and conclu-
sion (22) that diets formulated for adequate
amounts of energy and protein will not neces-
sarily provide adequate substrate for maximum
microbial growth. Hoover and Stokes (14) and
Stokes et al. (32) examined the relationship
1101
between bacterial protein yield and NSC:DIP
ratio and found that highest yields were as-
sociated with the narrowest ratio. In the pres-
ent trial, NSC:DIP ratios (DIP = RAP, esti-
mated in situ) were 3.1, 4.2, 3.0, and 3.8 for
the high RANSC and high RAP, high RANSC
and low RAP, low RANSC and high RAP, and
low RANSC and low RAP diets, respectively.
Intake of NSC was similar between the high
and low RANSC diets, but differences existed
in the amount of NSC digested in the rumen,
which affected bacterial yield, depending on
the amount of RAP in the diet. Therefore,
bacterial yield may be more appropriately ex-
pressed as a function of RANSC rather than
NSC.
Total tract apparent digestibility of N was
higher ( P < .01) for cows fed the high RANSC
diets than for those fed the low RANSC diets
(67.5 vs. 62.0%; Table 8). More NSC was
digested postruminally when cows were fed
the low RANSC diets (1.9 vs. 1.3 kg/d).

TABLE 8. Effect of amount of rumen-available nonstructural carbohydrate (RANSC) and rumen-available protein (RAP)
on intake, duodenal passage, and digestibility of N.'
High RANSC Low RANSC
Effect (P <)
High Low High Low
Item RAP RAP RAP RAP SEM C2 P3 C X
~~ ~ ~~ ~

N
Intake, g/d 700 691 755 722 17 .04 NSS NS
Passage to duodenum, g/d 555 580 566 648 32 NS .15 NS
Digested in rumen apparently, % 20.6 15.9 24.7 10.6 3.4 NS .03 NS
Digested in rumen truly, 7% 58.3 46.9 57.9 43.5 5.4 NS .04 NS
Total tract digestion, g/d 477 462 465 444 18 NS NS NS
% of N intake 68.3 66.9 61.9 62.1 1.3 .01 NS NS
Fecal NDF N, % of N 32.7 31.1 27.9 28.9 .9 .01 NS NS
Digestibility: % 89.6 89.7 89.5 89.2 .4 NS NS NS
NAN
Passage to duodenum, g/d 536 565 549 633 31 NS .12 NS
Bacterial N
Passage to duodenum, g/d 262 214 234 237 10 NS .04 .03
OMTD? g/kg 17.9 14.5 15.2 17.8 1.0 NS NS .01
NSCS digested, g/kg 36.7 29.3 35.5 42.7 2.7 .05 NS .03
'Least squares means.
2C = The RASNC effect.
3P = The RAP effect.
% x P = The interaction between RANSC and RAP.
5P > .IO.
6Total tract N digestibility corrected for NDF N (assuming that NDF N represents undigested feed N).
'The OM truly digested in the rumen.
8Nonstructural carbohydrate.

Journal of Dairy Science Vol. 76, No. 4, 1993

1102 ALDRICH ET AL.

Greater quantities of NSC may have been fer-
mented in the cecum, which would stimulate
microbial protein synthesis and, hence, fecal
output of N (23). When NDF N in feces was
assumed to represent undigested feed N [not
corrected for keratin (36)l and used to estimate
true digestibility, total tract digestibility of N
averaged 89.5% across treatments and was un-
affected by amount of RANSC or RAP (P >
.15).
Passage of AA t o the Small Intestine
The low RAP diets increased essential (P e
.Ol), nonessential (P < .06), and total AA (P e
.02) flows to the small intestine (Table 9). The
flow of Ile was lower (100 vs. 117 g/d) for
cows fed the low RAP diets (P e .Ol), and
flows of Arg and Met were similar across
treatments. The flows of all other individual
essential AA were higher when cows were fed
the low RAP diets. Amount of RANSC had no
effect on AA flow to the small intestine.
Generally, AA flows were lower than those
reported by McCarthy et al. (20) except for the
flow of Lys, which was slightly higher (252
and 260 vs. 246 and 228 gld) when cows were
fed the low RAP diets than when they were
fed diets supplemented with fish meal (20),
respectively. Blood meal supplied 29% of the
total CP in the low RAP diets. Lysine content
of blood meal is high relative to other protein
supplements (6) and, because of its low rumen
degradability (34), would be expected to de-
liver a substantial quantity of Lys to the small
intestine.
Schwab (29) determined that Lys was the
first-limiting AA for cows in peak and early
lactation when the proportional passage of Lys
to the small intestine was 12.2 to 12.6% of

TABLE 9. Effect of amount of rumen-available nonstructural carbohydrate (RANSC) and rumen-available protein (RAP)
on duodenal AA flow.
High RANSC Low RANSC
Effect ( P e)'
High Low High Low
AA RAP RAP RAP RAP SEM C2 P3

Essential
A% 132 132 128 139 4 NS4 NS
His 53 74 50 82 3 NS .01
Ile 120 96 113 104 4 NS .01
Leu 259 310 263 331 11 NS .01
LYS 212 252 201 260 7 NS .01
Met 48 49 47 50 2 NS NS
Phe 131 158 131 168 5 NS .01
Thr 142 153 139 161 5 NS .02
Val 153 184 149 192 5 NS .01
Total essential 1248 1408 1221 1485 45 NS .01
Nonessential
Ala 181 210 179 222 6 NS .01
ASP 296 318 286 330 10 NS .02
Glu 37 1 375 370 401 15 NS NS
GlY 169 181 179 182 9 NS NS
Pro 135 143 139 154 7 NS NS
Ser 142 155 139 162 5 NS .02
TYr 101 100 98 103 3 NS NS
Total nonessential 1395 1482 1389 1553 53 NS .06
Total AA 2643 2890 2610 3038 98 NS .02
lThe interaction between RANSC and RAP was not significant ( P > .lo).
2C = The RASNC effect.
3P = The RAP effect.
4P > .IO.

Journal of Dairy Science Vol. 76, No. 4, 1993

 NONSTRUCTURAL CARBOHYDRATE AND PROTEIN 1103
TABLE 10. Effect of amount of rumen-available nonstructural carbohydrate (RANSC) and rumen-available protein
(RAP) on milk production and composition.
High RANSC Low RANSC
Effect ( P c )
High Low High Low
Item RAP RAP RAP RAP SEM C1 P2 cx P3
Milk, kg/d 39.3 38.8 39.5 39.6 .7 NS4 NS NS
Milk fat, % 3.26 3.40 3.46 3.38 .07 NS NS NS
kg/d 1.26 1.31 1.36 1.34 .01 .01 NS .01
Milk protein, % 3.08 3.05 3.04 2.98 .02 .01 .05 NS
kg/d 1.20 1.17 1.19 1.18 .02 NS NS NS
4% FCM, kg/d 34.7 35.2 36.2 35.9 .3 .01 NS NS
SCC, x 1 @ / d 32.1 42.6 44.7 47.6 13.1 NS NS NS
IC = The RASNC effect.
*P = The RAP effect.
3C x P = The interaction between RANSC and RAP
4P > .IO.

essential AA flow. The low RAP diets and the
high RAP diets contained 17.5 to 17.9 and
16.5 to 17.0% Lys as a percentage of essential
AA flow, respectively. Methionine was
second-limiting or colimiting with Lys when it
constituted 3.8 to 3.9% of essential AA flow
(29). The low and high RAP diets contained
3.5 and 3.8% Met, respectively, as a percent-
age of essential AA flow. Based on those
criteria (29), Met may have been more limiting
than Lys for all diets.
Milk Production end Composition
Milk production averaged 39 kg/d across all
treatments and was unaffected (P > .lo) by
amount of RANSC or RAP (Table lo).
Production of 4% FCM was elevated by 1 kg/d
(P < .01) for cows fed the low RANSC diets.
Higher FCM for cows fed the low RANSC
diets resulted from marginally higher milk
production and fat percentages (P > .lo),
which resulted in higher fat yields (P < .Ol);
intakes of DM and NDF were higher when
cows were fed these diets and may have stimu-
lated higher FCM production. A RANSC by
RAP interaction (P < .01) existed on the yield
of milk fat, because milk fat percentage was
somewhat lower when the high RANSC and
high RAP and the low RANSC and low RAP
diets were fed.
Milk protein percentage was higher (P <
.01) when cows were fed the high RANSC
diets (3.07 vs. 3.01%) and when they were fed
the high RAP diets (3.06 vs. 3.02%; P < .OS).
An improvement in AA supply to the mam-
mary gland probably is not a plausible expla-
nation for the differences in milk protein per-
centages observed in the current study because
neither the high RANSC diets nor the high
RAP diets increased the flow of AA to the
small intestine. Increases in milk protein per-
centages with increasing RANSC or RAP may
have been related to slightly increased propi-
onate concentration in the rumen, which may
have increased glucose supply to the mammary
gland (7) or stimulated insulin production,
which, in turn, may have enhanced AA uptake
or protein synthesis by the mammary gland
(24).
The small but significant impacts of
RANSC and RAP on 4% FCM production and
composition may have reflected the marginal
but significant shifts in intake, rumen fermen-
tation end products, and nutrient flow to the
small intestine. The direction of these changes
varied by diet and may have been compensa-
tory in their effect on the milk production
variables. For example, intake increased when
cows were fed the low RANSC diets, but total
tract OM digestibility was lower. Improve-
ments in passage of bacterial N to the small
intestine when cows were fed the high RANSC
and high RAP diet were offset by decreased
passage of AA to the small intestine when the
high RAP diets were fed. Passage of Met and
Journal of Dairy Science Vol. 76, No. 4, 1993
1104 ALDRICH ET AL
Arg to the small intestine was not affected by
either RANSC or RAP and may have been the
limiting AA on all diets. Another reason for
the lack of major changes in milk production
resulting from differences in RANSC and RAP
might be that DMI was high enough on all
diets (25 kg/d) relative to milk production (39
kg/d), and adequate nutrients were available
for milk synthesis regardless of the amount of
RANSC or RAP.
CONCLUSIONS
A diet that contained 36% NSC, of which
80% was digested (high RANSC) in the ru-
men, and 17.5% CP, of which 66% (estimated
in situ; high RAP) was digested in the rumen,
resulted in the highest passage of bacterial N
to the small intestine. Decreasing either the
proportion of NSC or the proportion of protein
digested in the rumen reduced passage of bac-
terial N. Flow of AA to the small intestine was
increased when diets that contained less RAP
were fed, but these diets did not enhance milk
production or milk protein yield. Milk protein
percentage increased slightly, but significantly,
when cows were fed the high RANSC or high
RAP diets. Diets with elevated U P (low RAP)
probably were not advantageous because pas-
sage of bacterial N was lower, and those diets
failed to increase flows of Met, Arg, and Ile to
the small intestine. Overall, varying amounts
of RANSC and RAP did not dramatically in-
fluence performance in this short-term study.
ACKNOWLEDGMENTS
The authors thank Patrick Hillard and Terry
Cassidy for help with cow care and laboratory
work and gratefully acknowledge the financial
support of Nutrena Feeds (Elk River, MN),
Agway Inc. (Syracuse, NY) and Buzz Burhans,
Dairy Nutrition Associates, Inc. (Pawlet, VT).
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Journal of Dairy Science Vol. 76, No. 4, 1993