The Professional Animal Scientist 27 (2011):257–261

©2011 American Registry of Professional Animal Scientists

CstageS of maturity
: Effect of grazing,
ASE
at harvest,
TUDY

and glycerol treatment on wheat

harvested as silage
M. S. Gadberry,* PAS, D. Philipp,†1 P. A. Beck,‡ PAS, E. L. Brown,* and J. A. Hawkins*
*Cooperative Extension Service, University of Arkansas, Little Rock 72204;
†Animal Science Department, University of Arkansas, Fayetteville 72701; and
‡Southwest Research and Extension Center, Hope, AR 71801

ABSTRACT a slight improvement of IVDMD (P = With the emphasis on renewable

The objective of this project was to
examine the effect of grazing restric-
tion, stage of maturity at harvest, and
addition of glycerol to wheat (Triticum
aestivum L.) harvested as silage. The
project design was a split-split plot, and
the first treatment level was nongrazed
wheat compared with wheat grazed just
before first hollow stem. The secondary
treatment structure was harvesting the
wheat at anthesis versus soft dough ma-
turity. The third treatment structure was
applying glycerol to the wheat at 0, 5, 10,
or 15% of DM before ensiling. Nongrazed
wheat produced more yield than did
grazed wheat (P = 0.04), and harvesting
at soft dough produced more yield than
did harvesting at anthesis (P < 0.001).
The change in NDF, ADF, and IVDMD
was greater for nongrazed wheat from
anthesis to soft dough maturity compared
with grazed wheat (P ≤ 0.06). Graz-
ing had no effect on postensiling forage
chemical composition. Silage pH reached
acceptable levels (4.0 to 4.2) but was
not affected by plant maturity. Glycerol
addition, however, appeared to result in
1
Corresponding author: dphilipp@uark.edu
0.04).
Key words: glycerol, grazing, wheat
silage
INTRODUCTION
Beef cattle producers with farms in
Arkansas that consist predominately
of warm-season perennial grasses have
the option of no-till planting cool-
season annual grasses or small grains
in autumn to establish forages for
grazing in late winter and spring. For-
age not grazed during this period may
be used at a later time, but this is not
practical because the nutritive value
of these forages quickly declines, re-
sulting in unacceptable animal growth
rates (Anderson, 2006). In the case of
wheat (Triticum aestivum L.), grain
yields may also be affected whenever
the effect of grazing is not minimized
by removing animals before first
hollow stem (Redmon et al., 1995).
If grain production is not the goal,
producers have the opportunity to
use the wheat crop for silage, without
excessively compromising the length
of the grazing period.
energy, large-scale biodiesel facilities
may produce quantities of glycerol
that exceed demand for traditional
uses. As a result, glycerol has been
studied as a potential source of energy
in ruminant diets. Rémond et al.
(1993) reported that glycerol was
rapidly fermented in the rumen and
increased propionate proportions, but
DMI was decreased (DeFrain et al.,
2004). Coffey et al. (2008) indicated
that 10% of DM in the feed ration
may be the upper limit for glycerol
inclusion in the diet to avoid reduced
intake in cattle. If glycerol can be
used by anaerobic microbes, it also
seems reasonable to presume that
glycerol could be included in silages,
hence extending its possible uses. This
could potentially improve silage qual-
ity if fermentable carbohydrates had
diminished because of increased plant
maturity and fiber levels.
The authors are not aware of previ-
ous studies in which glycerol was used
as a silage additive. Therefore, the
objectives of this study were to ex-
amine the effect of grazing restriction
and stage of plant maturity of wheat
on crop yield and evaluate the effects
of glycerol addition on changes in
258
chemical composition of wheat silage
preensiling and postensiling.
MATERIALS AND METHODS
Site Description
This project was conducted as an
on-farm producer demonstration in
central Arkansas (35°03′ N, 92°33′ W).
The farm site consisted of Gallion silt
loam and Roxana fine sandy loam soil
with 0 to 1% slope (NRCS, 2010).
On October 7, 2007, a locally avail-
able pasture wheat variety (Amazin
Grazin, Agri Coop Supply, Morrilton,
AR) was no-till drilled into 4.5 ha of
an existing bermudagrass [Cynodon
dactylon (L.) Pers.] sod of unknown
variety. Beginning on November 27,
2007, 53 fall calves were weaned onto
the site. In addition to the research
site of no-till drilled wheat, calves
had continuous access to an ad-
ditional 18 ha of wheat (var. Coker
9663, Agri Coop Supply, Conway,
AR) that had been broadcast after
disking on October 7, 2007, (fertil-
ized with N at 135 kg/ha and K2O at
100 kg/ha) and 38.5 ha of fallow crop
ground that produced winter annual
grasses (Bromus spp.) and volunteer
annual ryegrass (Lolium multiflorum
Lam.). Because this project involved
observing the influence of grazing on
a producer farm in which the research
leaders had no control over cattle
management, there was no animal
care committee review of the project.
By observation, it appeared the farm
was implementing industry-accepted
beef cattle management practices
regarding animal health, performance,
and value improvement, such as de-
horning and castrating. Observations
of calf body condition while collect-
ing forage data suggested that these
cattle were not malnourished during
any phase of the study.
Before fall grazing, 3 grazing
exclosures measuring 2.4 × 4.9 m
were assembled on the no-till drilled
paddock. On February 28, 2008, a
second set of grazing exclosures with
the same dimensions was assembled
adjacent to the original 3 exclosures
to simulate calf removal just before
Gadberry et al.
first hollow stem. After the assembly
of the second grazing exclosure, 67
kg/ha of N and 100 kg/ha of K2O
equivalent rates were applied to the
fall and spring restriction areas.
Forage Samples and Analyses
Forage was harvested from approxi-
mately one-half of each of the 6 plots
(grazing exclosures) on April 22, 2008,
(anthesis) and the remaining one-
half the following May 7 (soft dough)
and weighed for yield determination.
The samples were then transferred to
facilities at the University of Arkan-
sas, Animal Science Department,
Fayetteville. Samples were placed
on a dry, clean concrete surface in a
covered barn and shredded using a
commercial lawn mower (Honda HRX
2172, Alpharetta, GA). Material was
collected in a bag attached to the
mower. Average particle length after
shredding was 1.8 cm.
Random samples from all treat-
ments were then monitored for
moisture concentration by repeatedly
placing subsamples in a microwave
oven to evaporate moisture. Dry mat-
ter was calculated by subtracting final
from initial weight. After the samples
reached a DM concentration of 40%,
large plastic bags were filled with
bulk samples, closed, and immedi-
ately transferred to the laboratory
for further processing. Samples were
prepared by adding predetermined
amounts of glycerol (0, 5, 10, and 15%
of DM) to the original sample mate-
rial. Target weight for each particular
sample was 500 g. Sample material
was weighed and thoroughly mixed in
large plastic buckets with the assigned
amounts of glycerol before mate-
rial was transferred to plastic bags
(Applica Consumer Products, Miami
Lakes, FL), cut to size, and closed
with a vacuum sealer (FoodSaver,
Signature series V 825, Tilia Inc., San
Francisco, CA). Sealed samples were
wrapped again with another bag and
vacuum sealed to prevent rupture and
to ensure near-anaerobic conditions
throughout the experiment. Samples
were then incubated at 20°C in dark-
ness for 45 d.
After the 45-d ensilage period, DM
concentration was determined by sep-
arating approximately 100 g of fresh
sample and drying in a forced-air
drying oven at 55°C until a constant
weight was achieved. Furthermore, the
pH of fresh samples was measured im-
mediately upon opening the ensilage
bags. For this, 30 g of fresh sample
was mixed with 300 mL of deionized
water and stirred occasionally until
pH was measured after 30 min had
elapsed. Remaining sample material
was transferred to a Ziploc bag (S.C.
Johnson and Son Inc., Racine, WI)
and stored at −15°C. Samples previ-
ously used for determination of DM
content were ground in a Wiley mill
(Thomas Scientific, Swedesboro, NJ)
to pass a 1-mm screen and stored at
room temperature until further chemi-
cal analysis.
Neutral detergent fiber and ADF
were determined according to the
procedures described by Van Soest
(1963) and Goering and Van Soest
(1970). Crude protein was determined
according to AOAC (1995).
In vitro DM disappearance was
determined by batch procedures out-
lined by ANKOM Technology Corp.
(Fairport, NY). Rumen fluid was
obtained from a ruminally cannulated
crossbred steer that was offered a
diet of 80% alfalfa (Medicago sativa
L.) and 20% concentrate at a main-
tenance level of intake. The steer was
adapted to the diet for 10 d before
the rumen fluid was collected.
Volatile fatty acids were analyzed
using an automated Hewlett Packard
5890 gas chromatograph (Hewlett
Packard Co., Palo Alto, CA) fitted
with a Nukol fused silica capillary col-
umn (30 m × 0.25 mm i.d. × 0.25 μm
film thickness; Supelco Inc., Bellefon-
te, PA), a 5 m × 0.25 mm i.d. fused
silica intermediate polarity guard
column (Supelco Inc.), and an flame
ionization detector. Chromatographic
peaks were qualified by comparing
retention times of sample compo-
nent peaks to those of a volatile acid
standard mix (Supelco Inc.). Fatty
acid amounts were quantified by using
a known amount of 2-ethyl butyrate
as an internal standard within each

Table 1. Effect of grazing restriction and stage of wheat plant maturity on yield and preensiling chemical
composition
Nongrazed
Item Anthesis
Yield, kg/ha 2,035
Moisture, % 60.4
CP, % 11.0
NDF, % 63.3a
ADF, % 33.9a
IVDMD, % 70.6b
a–c
Least squares means within a row with different superscripts differ (P < 0.05).
sample. Lactic acid concentrations
were determined according to proce-
dures described by Gloster and Harris
(1962). In addition, the lactate-to-
acetate ratio and total organic acid
concentration were calculated.
Statistical Analyses
Forage yield and preensiling chemi-
cal composition were analyzed as
a split plot within a randomized
complete block design. The whole-plot
unit was grazed versus nongrazed and
the subplot unit was anthesis versus
soft dough stage of maturity. Forage
quality was initially analyzed as a
split-split plot within a randomized
complete block design; however, the
effect of grazing or any interaction
with grazing was not significant (P >
0.05) for any of the forage chemical
composition measures. As a result,
forage chemical composition data
were analyzed as a split plot within
a randomized complete block. The
whole-plot unit was stage of matu-
rity and the subplot unit was level of
glycerol addition before ensiling. Data
analysis was completed using PROC
MIXED (SAS Inst. Inc., Cary, NC).
RESULTS AND DISCUSSION
Preensiling Responses
The response of yield and forage
chemical composition before ensiling
is presented in Table 1. Forage yield
was affected independently by graz-
Influence of grazing and glycerol addition on wheat silage
Grazed
Soft dough Anthesis Soft dough
3,408 1,709 2,672
50.4 62.9 50.1
10.9 12.3 11.7
56.9c 59.9b 57.4c
30.7b 31.3b 31.0b
74.9a 72.8a 73.6a
ing (P = 0.04) and stage of maturity
at harvest (P < 0.001). Grazed wheat
and removal before first hollow stem
produced on average 530 kg less DM
per hectare in comparison with wheat
that was never grazed. However, aver-
aged across the 2 stages of maturity,
the grazed wheat still produced more
than 2,000 kg/ha of harvested for-
age. The initial forage harvest oc-
curred during anthesis, whereas the
second harvest occurred 2 wk later
when wheat was in soft dough stage.
Averaged across grazing treatments,
this 2-wk delay in harvest resulted
in a 1,168-kg increase in forage per
hectare.
Forage moisture at ensiling was not
affected by grazing (P = 0.69) or the
interaction of grazing with stage of
maturity (P = 0.60); however, the
moisture content of wheat harvested
at anthesis contained 62% moisture at
ensiling compared with 50% moisture
at ensiling for wheat harvested at
soft dough (P < 0.01). Both levels of
moisture were acceptable for ensil-
ing forages where 55% moisture is a
commonly accepted target for grass
silages.
None of the forage chemical con-
stituents measured were affected
independently by grazing. However,
an interaction between grazing restric-
tion and stage of maturity at harvest
was detected for NDF (P = 0.03),
ADF (P = 0.06), and IVDMD (P <
0.01). The change in quality charac-
teristics measured from anthesis to
soft dough was greater in nongrazed
259
P-value
Graze ×
SE Graze Maturity maturity
281.5 0.04 <0.001 0.14
2.38 0.69 <0.01 0.60
0.60 0.19 0.55 0.64
1.07 0.13 <0.01 0.03
1.02 0.18 0.04 0.06
0.38 0.18 0.001 <0.01
forages compared with grazed forages.
For instance, the NDF content was
6.4 percentage units lower at the soft
dough stage for nongrazed wheat but
only 2.5 percentage units lower for
grazed wheat. Harvesting at the soft
dough stage resulted in a significant
(P < 0.05) reduction in NDF and
ADF, whereas IVDMD increased.
Crude protein was not affected by
stage of maturity (P = 0.55) or its
interaction with grazing restriction
date (P = 0.64).
Several authors have addressed the
effect of maturity on ruminal digest-
ibility. Beck et al. (2009) reported
that maturity of wheat forage at har-
vest, preservation method, and their
interaction did not affect (P ≥ 0.15)
ADG in calves when wheat was fed as
20 or 40% of the diet. Furthermore,
these diets did not affect (P ≥ 0.13)
passage rate, ruminal retention time,
or fecal output. However, DM and
NDF digestibility of 40% boot-stage
wheat forage were greater (P < 0.01)
than those of diets containing forage
harvested in dough stage. There is
sufficient evidence that digestibility
decreases with increasing plant matu-
rity. Coblentz et al. (2000) reported a
reduction in DM disappearance of 14
percentage units from boot to dough
stage in wheat forage.
In our study, IVDMD increased by
4.3 percentage units preensiling under
a nongrazed treatment. It is not clear
why IVDMD slightly increased in this
situation, especially given opposing
evidence and the general assumption
260 Gadberry et al.

Table 2. Effects of plant maturity and level of glycerol addition1 on nutritive value (in % of DM) in wheat after 45
d of ensilage in vacuum-sealed plastic bags

Anthesis Soft dough P-value

Item 0 5 10 15 0 5 10 15 SE Maturity Glycerol Maturity × glycerol

Moisture, % 65.0 63.6 62.4 61.6 52.2 51.0 50.8 50.5 1.78 0.03 0.08 0.80
CP, % 11.1 10.6 10.2 10.7 11.8 11.6 11.4 11.4 0.73 0.44 0.42 0.85
NDF, % 63.8 61.7 59.5 58.9 58.7 57.0 54.5 53.7 1.02 <0.01 <0.001 0.99
ADF, % 36.1 35.8 33.9 33.8 34.0 32.6 31.6 30.9 0.89 0.01 <0.001 0.59
IVDMD, % 80.8 82.4 83.7 83.6 81.4 82.5 82.5 82.2 0.96 0.66 0.04 0.43
1
Glycerol was added at levels of 0, 5, 10, and 15% of DM for each sample.

that plant tissue digestibility de-
creases with increasing maturity. One
possible explanation may be that as
the digestibility of the wheat inflores-
cence should increase between anthe-
sis and soft dough stages, so should
the whole-plant digestibility, given the
relatively high inflorescence-to-stem
weight ratio during the grain-filling
phase. Zebrowski (1999) reported that
in late milk stage, the wheat inflores-
cence accounts for 85% of the weight
of stem plus inflorescence. These
authors removed the leaves before re-
cording the weights, but other authors
(Siddique et al., 1989) reported a
sharp increase between terminal spike-
let and anthesis, with weight ratios
changing from 20 to 50%. However, it
is not clear why differences of IVD-
MD observed under nongrazed wheat
in our study were not present under
the grazed treatment, unless grazing
influenced plant maturity, altering
the IVDMD response. As previously
noted, fiber measures were lower and
CP numerically greater in the grazed
wheat.
Postensiling Responses
The results are summarized in
Tables 2 and 3. Grazing had no effect
on any postensiling chemical constitu-
ents measured and was removed from
the postensiling chemical composi-
tion analysis. The effect of preensil-
ing moisture differences resulted in
a significant difference (P = 0.03) in
postensiling moisture level between
the 2 stages of maturity at harvest.
Moisture level tended to differ (P =
0.08) by level of glycerol addition and
averaged 58.6, 57.3, 56.6, and 56.0%
moisture for 0, 5, 10, and 15% glyc-
erol, respectively. Crude protein was
not affected by maturity (P = 0.44),
glycerol addition (P = 0.42), or their
interaction (P = 0.85). The effect
of maturity on NDF (P < 0.01) and
ADF (P = 0.01) remained significant
after preservation. The addition of
glycerol affected (P < 0.001) NDF
and ADF concentrations. This in
part can be attributed to the fact
that glycerol does not contain struc-
tural carbohydrates and any addition
should result in a reduction in mea-
sures of plant fiber. However, calcu-
lated levels of NDF and ADF at dif-
ferent levels of glycerol addition were

Table 3. Effects of plant maturity and level of glycerol addition1 on organic acid composition in wheat after 45 d
of ensilage in vacuum-sealed plastic bags

Anthesis Soft dough P-value

Maturity ×
Item 0 5 10 15 0 5 10 15 SE Maturity Glycerol glycerol
pH 4.2 4.2 4.1 4.1 4.0 4.0 4.0 4.0 0.05 0.12 0.28 0.42
Acetate, Mmol/L 11.4b 10.3b 10.0b 11.7b 11.4b 14.3a 13.9a 11.9b 1.74 0.18 0.73 0.01
Propionate, 0.12c 0.16bc 0.14bc 0.12c 0.20ab 0.14bc 0.18bc 0.22a 0.06 0.30 0.81 <0.01
Mmol/L
Butyrate, Mmol/L 0.09 0.83 0.10 0.04 0.65 0.18 0.00 0.01 0.32 0.86 0.26 0.24
Lactate, Mmol/L 25.9 24.6 23.9 23.8 24.8 22.4 24.2 23.5 1.94 0.66 0.72 0.91
Lactate:acetate 2.4 2.5 2.5 2.2 2.4 1.6 1.8 2.1 0.39 0.27 0.39 0.10
Lactate, % of 68.9 68.8 70.0 66.4 66.9 59.7 62.7 65.3 4.86 0.31 0.63 0.40
total acids
a–c
Least squares means within a row with different superscripts differ (P < 0.10).
1
Glycerol was added at levels of 0, 5, 10, and 15% of DM for each sample.
 Influence of grazing and glycerol addition on wheat silage
lower than actual measures. In vitro
DM disappearance was singularly af-
fected (P = 0.05) by the addition of
glycerol. Average IVDMD across the
2 maturity stages was 81.1, 82.4, 83.1,
and 82.9 for 0, 5, 10, and 15% glyc-
erol, respectively. Neither organic acid
concentration nor pH was affected
independently by either stage of ma-
turity or glycerol addition. However,
acetate (P = 0.01), propionate (P <
0.01), and lactate-to-acetate ratio (P
= 0.10) were affected or tended to be
affected by the level of glycerol within
stage of maturity.
In general, data regarding the ef-
fects of glycerol as a silage additive
on forage chemical composition are
scarce. Therefore, it is difficult to
compare our results with previous
work and determine whether other
authors observed similar or contradic-
tory results. Based on our data, the
addition of glycerol may not strongly
influence the chemical composition of
silage.
IMPLICATIONS
Glycerol may become a readily
available by-product from the biodies-
el industry and could serve as a silage
additive. Our results suggested that
glycerol can be added to wheat silage
without negative effects regarding di-
gestibility, fiber content, and organic
acid composition. Furthermore, delay-
ing wheat harvest from anthesis to
soft dough may not result in reduced
nutritive value. However, field-scale
research including an economic analy-
sis is needed to test palatability of
glycerol-treated silage and resulting
animal performance.
ACKNOWLEDGMENTS
This project was funded by Cooper-
ative State Research, Education, and
Extension Service, Agriculture and
Food Research Initiative (National
Research Initiative) grant agree-
ment 2006-55101-17440. The authors
express appreciation to Schafer farms
(Toad Suck, AR) for the demonstra-
tion site.
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