Archives of Animal Nutrition

ISSN: 1745-039X (Print) 1477-2817 (Online) Journal homepage: http://www.tandfonline.com/loi/gaan20

Effects of the replacement of concentrate and

fibre-rich hay by high-quality hay on chewing,
rumination and nutrient digestibility in nonlactating Holstein cows
Maria-Theresia Kleefisch, Qendrim Zebeli, Elke Humer, Iris Krger, Paul Ertl
& Fenja Klevenhusen
To cite this article: Maria-Theresia Kleefisch, Qendrim Zebeli, Elke Humer, Iris Krger, Paul Ertl
& Fenja Klevenhusen (2016): Effects of the replacement of concentrate and fibre-rich hay by
high-quality hay on chewing, rumination and nutrient digestibility in non-lactating Holstein
cows, Archives of Animal Nutrition, DOI: 10.1080/1745039X.2016.1253227
To link to this article: http://dx.doi.org/10.1080/1745039X.2016.1253227

Published online: 18 Nov 2016.

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Date: 23 November 2016, At: 03:42

ARCHIVES OF ANIMAL NUTRITION, 2016

http://dx.doi.org/10.1080/1745039X.2016.1253227

Eects of the replacement of concentrate and bre-rich hay

by high-quality hay on chewing, rumination and nutrient
digestibility in non-lactating Holstein cows
Maria-Theresia Kleescha, Qendrim Zebelia, Elke Humera, Iris Krgera, Paul Ertlb
and Fenja Klevenhusena
a

Institute of Animal Nutrition and Functional Plant Compounds, Department for Farm Animals and
Veterinary Public Health, University of Veterinary Medicine Vienna, Vienna, Austria; bInstitute of Livestock
Sciences, Department of Sustainable Agricultural Systems, BOKUUniversity of Natural Resources and Life
Sciences Vienna, Vienna, Austria
ARTICLE HISTORY

ABSTRACT

The aim of this study was to investigate the eects of high-quality

hay with an elevated sugar content alone or with graded amounts
of concentrate feed on chewing and ruminating activity, apparent
total tract digestibility (ATTD) and ruminal pH at dierent time
points after feeding in the free ruminal liquid (FRL) and the particle-associated ruminal liquid (PARL). Eight rumen cannulated nonlactating Holstein cows were arranged in a Latin square design in
four experimental runs lasting 25 d each. The four diets tested
were 60NQ (60% normal-quality hay + 40% concentrate), 60HQ
(60% high-quality hay + 40% concentrate), 75HQ (75% high-quality hay + 25% concentrate) and 100HQ (100% high-quality hay).
Normal and high-quality hays diered in sugar contents (11.3% vs.
18.7% in dry matter [DM]), neutral detergent bre (NDF; 57.7% vs.
46.3% in DM), acid detergent bre (ADF, 35.0% vs. 23.5% in DM)
and crude protein (CP, 11.3% vs. 23.5% in DM). Data showed that
ATTD of DM, CP, NDF and ADF was higher with the high-quality
hay diets. Time spent eating was reduced with high-quality hay.
However, time spent ruminating was longest in Group 100HQ. In
all groups, ruminal pH of FRL and PARL decreased with time after
the morning feeding. But 10 h later, pH of Group 100HQ was
higher again compared with the other groups. Considering the
average pH in FRL over all measured time points, cows in Groups
60NQ and 100HQ had higher pH values of 6.85 and 6.83, respectively. Regarding pH values in PARL, animals of Group 60NQ displayed the highest pH value (6.68), whereas the lowest value of
6.21 was found in Group 60HQ. Overall, results suggest that highquality hay maintains the diets structural eectiveness by stimulating rumination and stabilising ruminal pH while greatly improving ATTD. However, the structural eectiveness of the high-quality
hay gets impaired with increasing proportion of concentrate feed
in the diet.

CONTACT Fenja Klevenhusen

fenja.klevenhusen@vetmeduni.ac.at

2016 Informa UK Limited, trading as Taylor & Francis Group

Received 20 June 2016

Accepted 4 October 2016
KEYWORDS

Concentrates; cows;
digestibility; bre content;
forage; hay; mastication; pH;
rumination

M.-T. KLEEFISCH ET AL.

1. Introduction
Concentrate feeds have long become an important part of dairy cow diets to meet their
high demands in energy and nutrients for milk production. However, commonly used
concentrate feeds largely contain human edible products like grains, thus their production and feeding to livestock competes with the human food availability (Ertl et al.
2016). Additionally, feeding starch-rich concentrate feeds like grains in large amounts
at the expense of bre-rich feeding concentrates poses risks for the cows health due to
their fast ruminal fermentation and acidogenic potential (Zebeli, Dijkstra, et al. 2008).
A viable and reasonable alternative to concentrate feeding in dairy cattle might be
the feeding of high-quality hay, i.e. a roughage rich in crude protein (CP) and energy in
form of water soluble carbohydrates that could ideally maintain milk production and at
the same time support ruminal health. High-quality hay is grown, harvested and dried
under favourable conditions. It is rich in energy due to its high contents of sugars that
can reach 20% and more of DM.
Some studies have shown the benecial eects of increased levels of sugars in forages
on animal production and animal nitrogen (N) use eciency (Lee et al. 2001; Merry
et al. 2006). However, the structural eectiveness of high-quality hay is likely to be
reduced due to lower bre (Tafaj et al. 2005) and higher content of sugars with high
ruminal degradability that may increase the acidogenic properties of the diet (Oba
2011). The structural eectiveness of forages in ruminants is determined by their
capability to induce intensive chewing and maintain proper rumen conditions, mainly
optimal pH for rumen functioning through saliva buering (Mertens 1997). The extent
to which such high-quality hay can be used in dairy cow rations to replace normal brerich hay to improve the nutritional value of the diet without impairing its structural
properties is of high interest with regard to ruminant health. The interaction of highquality hay with starchy concentrates has rarely been investigated (Tafaj et al. 2005);
thus, we were especially interested in possible synergistic eects of high-quality hay rich
in sugar and concentrate feed rich in starch on the ruminal pH.
Our hypotheses in this study were that high-quality hay with elevated sugar content
still provides enough structural eectiveness to induce a sucient chewing activity and
maintain ruminal pH. However, we assumed that adding concentrate feed as part of the
diet is likely to reduce the structural eectiveness and increases the risk for rumen
acidotic conditions due to increased amounts of easily fermentable carbohydrates in the
diet.
Therefore, the aim of this study was to evaluate the eects of feeding high-quality
hay alone or with graded amounts of concentrate feed on chewing activity, ruminal pH
of particle-associated ruminal liquid (PARL) and free ruminal liquid (FRL) and apparent total tract digestibility (ATTD) in a model feeding experiment with rumen cannulated non-lactating Holstein cows.

2. Materials and methods

2.1. Animals and diets
All procedures involving animal handling and treatment were approved by the institutional ethics committee of the University of Veterinary Medicine Vienna, Austria and

ARCHIVES OF ANIMAL NUTRITION

the national authority according to 26 of the Law for Animal Experiments,

Tierversuchsgesetz 2012, Bundesgesetzblatt I Nr. 114/2012 (permission of animal
experiment: Bundesministerium fr Wissenschaft, Forschung und Wirtschaft
68.205/0104 WF/II/3b/2014).
The trial was conducted at the Dairy Research Facilities in Kremesberg of the
University of Veterinary Medicine Vienna, Austria. Eight ruminally cannulated
(100 mm i.d.; Bar Diamond, Parma, ID) non-lactating and non-pregnant Holstein
cows (initial BW: 851 75 kg, mean SD) were used for the trial to enable frequent
sampling of rumen digesta. One week before starting the experiment, all cows were fed
common bre-rich hay only ad libitum and adapted to the feeding troughs. The cows
were kept in one group in a loose-housing stable with straw bedding and were arranged
in a double 4 4 Latin square design, balanced for carry-over eects. The experiment
was conducted in four consecutive runs, each run lasting 25 d, with the rst 6 d of
stepwise increasing the concentrate proportion in the concentrate fed groups starting
with 15% of dietary DM, followed by a 19-d long feeding period. Three of the four
dietary treatment groups contained the high-quality hay, with either 0% concentrate
(100HQ), 25% concentrate (75HQ) or 40% concentrate (60HQ). The fourth group
(60NQ), considered as control treatment, was fed with 60% normal bre-rich hay and
40% concentrate (Table 1). The bre-rich hay (second cut meadow hay beginning of
blooming; approximately 40% Dactylis glomerata, 30% other grasses, including Festuca
pratensis, Alopecurus pratensis and Arrhenatherum elatius, 20% clover and 10% herbs)
contained per kg DM 113 g sugars, 577 g neutral detergent bre (NDF), 350 g acid
Table 1. Ingredients and chemical composition of the experimental diets.
Experimental diets*
Diet composition [g/kg DM]
Normal hay
High-quality hay
Concentrates
Concentrate composition [g/kg DM]
Barley grain
Dried beet pulp
Rapeseed meal
Calcium carbonate
NaCl
Mineralvitamin premix
Urea
Chemical composition [g/kg DM]
Organic matter
Crude protein
Crude bre
Neutral detergent bre
Acid detergent bre
Non-bre carbohydrates
Nitrogen-free extractives
Starch
Sugars
Ether extract
Net energy lactation [MJ]

60NQ

60HQ

75HQ

100HQ

600

400

600
400

750
250

1000

730

221
5.00
4.00
20.00
20.00

745
139
90.0
4.00
2.00
20.00

745
139
90.0
4.00
2.00
20.00

921
159
212
424
245
330
539
182
84.2
10.7
6.20

925
192
159
359
180
360
558
178
129
16.0
7.24

921
208
180
398
201
300
518
111
150
14.8
7.11

915
235
213
463
235
200
454
0.00
187
12.8
6.89

Notes: *60NQ, 60% normal hay + 40% concentrate; 60HQ, 60% high-quality hay + 40% concentrate; 75HQ, 75% highquality hay + 25% concentrate; 100HQ, 100% high-quality hay; contained per kg: 190 g Ca, 50 g P, 30 g Mg, 90 g Na,
1.2 g Cu, 6 g Zn, 4 g Mn, 0.05 g Se, 0.05 g Co, 0.35 g I, 2.5 g vitamin E, 800,000 IU vitamin A, 80,000 IU vitamin D3.

M.-T. KLEEFISCH ET AL.

detergent bre (ADF), 35.3 g acid detergent lignin (ADL) and 113 g CP; whereas the
high-quality hay (Lolium perenne as main grass, mixed equal proportions of 1st and 2nd
cut, beginning of ear emergence) contained per kg DM 187 g sugars, 463 g NDF, 235 g
ADF, 14.8 g ADL and 235 g CP. Both hays had a similar stem length of 1012 cm. As
the CP content of the high-quality hay was very high, two dierent concentrate
mixtures in the same meal form were formulated having equal amounts of sugars
and starch to adjust the ratio of energy to nutrients among diets 60NQ and 60HQ.
Concentrate feed of diets 60HQ and 75HQ contained per kg DM 128 g CP, 59.2 g ash,
203 g NDF, 97.8 g ADF, 17.9 g ADL, 445 g starch and 41.0 g sugars. Concentrate feed of
diet 60NQ contained per kg DM 228 g CP, 61.0 g ash, 196 g NDF, 89.0 g ADF, 10.3 g
ADL, 454 g starch and 41.0 g sugars. Although total NDF was similar between the two
concentrates, beet pulp, rich in soluble NDF, was included in the concentrate for the
high-quality diets at the expense of rapeseed meal to provide less protein and to reduce
the concentrates disguising eect on forage NDF structural eectiveness.
Cows had free access to hays oered in separate feeding troughs (two troughs with
normal-quality hay and four troughs with high-quality hay), which were equipped with
electronic scales and computer-regulated access gates (Insentec B.V., Marknesse, The
Netherlands), enabling accurate electronic recording of the individual hay intake after
each visit to the trough. Fresh hay was continuously provided during the day. In
addition, the cows had access to concentrates starting from 10:00 h in a computercontrolled feeding station (Delaval GesmbH, Eugendorf, Austria), which enabled controlled intake of the concentrates for each cow. The daily allowance of concentrates was
set based on intake of hay of the previous day in order to maintain the required
concentrate to hay ratio for each cow. Free access to water and mineral stones (H.
Wilhelm Schaumann GmbH & Co. KG, Baden, Austria) was constantly provided.
2.2. Eating and chewing activity
Daily feed intake was recorded automatically by individual cow transponders.
Rumination and eating activities were recorded using noseband sensors (RumiWatch
System, ITIN + HOCH GmbH, Ftterungstechnik, Liestal, Switzerland) over four
consecutive days for each cow at the beginning of the feeding period. The system is
integrated in a halter and consists of a vegetable oil-lled silicone tube connected to a
pressure sensor, which records pressure changes in the oil-lled tube caused by the
cows jaw movements at a frequency of 10 Hz. Raw data were stored on the internal
storage device for the whole 4-d recording period, then downloaded via universal serial
bus on a computer and analysed using RumiWatch converter 0.7.3.11 (ITIN + HOCH
GmbH, Ftterungstechnik, Liestal, Switzerland).
2.3. Sampling of ruminal uid
To obtain ruminal uid over the course of 10 h, samples were collected from each cow
on 2 d, between d 13 and 18 of each experimental run. On the rst day, samples were
obtained before the morning feeding at 06:00 h (0 h), 4 and 8 h after the morning
feeding and on the second day, 2, 6 and 10 h after the morning feeding, respectively.
First, solid rumen digesta (approximately 200 g) was manually taken from the dorsal

ARCHIVES OF ANIMAL NUTRITION

rumen sac and used to obtain the PARL fraction of the ruminal liquid, using the
technique described earlier (Zebeli, Tafaj, et al. 2008). FRL was taken using an aspiration tube (Ruminator; T. Geishauser, Wittibreut, Germany) from the ventral rumen sac.
The pH was immediately measured using a pH electrode (Mettler Toledo SevenGo
Portable pH Meter SG2; Mettler-Toledo GmbH, Schwerzenbach, Switzerland) which
was calibrated with buers of pH 4.0 and 7.0.
2.4. Determination of ATTD of nutrients
To determine the ATTD, each cow received pre-weighed 20 g of the external marker TiO2
twice daily at 08:00 and 17:00 h through the rumen cannula for the last 9 consecutive days of
each run. During the last 7 days of each run, 200 g fresh faecal grab samples were collected
rectally of each cow at 09:00 and 17:30 h and stored at 4C. After the last day of sampling,
faeces were pooled by animal, mixed thoroughly and 1000 g were freeze-dried and ground
through a 0.5-mm screen (Ultra Centrifugal Mill ZM 200, Retsch, Haan, Germany). Dry
matter (DM) was determined by oven drying at 100C for 4 h. The approach for the
photometrical determination of the TiO2 concentration followed the specications of
Brandt and Allam (1987) and the modication of Glindemann et al. (2009).
2.5. Nutrient analyses
Feed samples were collected weekly. Samples were dried at 65C in a forced-air oven for
48 h. Afterwards, the samples were ground through a 0.5-mm screen (Ultra Centrifugal
Mill ZM 200, Retsch, Haan, Germany) before analyses. DM was determined by oven
drying at 100C for 24 h. Ash was analysed by combustion in a mue furnace over
night at 580C. Organic matter (OM) was calculated as
OM g=kg DM 1000 g=kg DM  Ash g=kg DM:
NDF, ADF and ADL were determined exclusive of residual ash following the ocial
analytical methods of (VDLUFA 1997 [1976]) using the Fibre Therm FT 12 (Gerhardt
GmbH & Co. KG, Knigswinter, Germany) with heat-stable -amylase in case of NDF.
Amount of non-brous carbohydrates (NFC) was calculated as (all nutrients as g/
kg DM):
NFC g=kg DM 1000  NDF CP Ether extract EE Ash:
The amount of nitrogen-free extractives (NfE) was calculated as (all nutrients as g/
kg DM):
NfE g=kg DM OM  EE CP Crude fibreCF:
Crude bre (CF)was analysed according to the ocial analytic method of VDLUFA
(19882012 [1976]). CP was analysed following the Kjeldahl method (VDLUFA 2007
[1976]) and EE using the Soxhlet extraction system (Extraction System B-811, Bchi,
Flawil, Switzerland). Starch in concentrates was analysed polarimetrically after treatment with hydrochloric acid. Sugar in hays and concentrates was determined by

M.-T. KLEEFISCH ET AL.

analysing the amount of reduced sugars after inversion following the method of LuSchoorl (VDLUFA 2012 [1976]). Net energy for lactation (NEL) was calculated with
NEL values for concentrate feedstus reported in DLG Futterwerttabellen (1997).
Values for NEL from the two hays were estimated according to DLG (2011/2013) as
NEL MJ=kg DM 0:61 0:004q  57  Metabolisable energy ME MJ=kg DM;
where q = {ME [MJ/kg DM]/Gross energy (GE) [MJ/kg DM]} 100.
Therefore, GE in hay was calculated as
GE MJ=kg DM 0:0239  CP g=kg DM 0:0398  EE g=kg DM 0:0201  CF g=kg DM
0:0175  NfE g=kg DM DLG 2013

and the ME was calculated according to GfE (1998) as (all fractions as g/kg DM):
ME MJ=kg DM 13:69  0:01624  CF 0:00693  CP  0:0067  Ash:

2.6. Statistical analyses

Statistical analyses were performed using the MIXED procedure of SAS (version 9.4;
SAS Institute Inc., Cary, NC, USA). The linear mixed model for feed intake and
chewing and ruminating activity is shown as follows:
Yijkl Si RSij Tk Dl eijkl
where Yijkl is the dependent variable, is the overall mean, Si is the xed eect of square
(i, 12), R(Sij) is the xed eect of the jth run (j, 14) within square, Tk is the xed eect
of the kth diet (k, 14), Dl is the xed eect of the lth day (for feed intake l, 119; for
chewing and ruminating l, 14) considered as repeated measures. Dierent variance
covariance matrix structures (auto-regressive 1, unstructured, compound symmetry) of
repeated measures were separately modelled for each variable and the covariance
structure with the smallest Bayesian information criterion (BIC) was used (Littell
et al. 2000). The eijkl is the residual error assumed to be normally distributed.
For ATTD, the linear mixed model was
Yijkl Si RSij Tk CSli eijkl
where Yijkl is the dependent variable, is the overall mean, Si is the xed eect of square
(i, 12), R(Sij) is the xed eect of the jth run (j, 14) nested within square, Tk is the
xed eect of the kth diet (k, 14), C(Sli) is the random eect of the lth cow within the
ith square and eijkl is the residual error assumed to be normally distributed.
The linear mixed model for both pH in FRL and PARL is shown as follows:
Yijklmn Si RSij Tk DH lm TDH klm CSni eijklmn
where Yijklmn is the dependent variable, is the overall mean, Si is the xed eect of
square (i, 12), R(S)ij is the xed eect of the jth run (j, 14) nested within square, Tk
is the xed eect of the kth diet (k, 14), D(H)lm is the xed eect of the lth day D (l,

ARCHIVES OF ANIMAL NUTRITION

12) nested within the mth time (m, 010), TD(H)klm is the xed eect of the
interaction between the kth diet and lth day within the mth time and C(S)nj is the
random eect of the nth cow within the ith square. And eijklmn is the residual error
assumed to be normally distributed.
Multiple comparisons among treatments were evaluated by Tukeys test. The signicance level was set at p 0.05 and a trend was considered when 0.05 < p 0.10. In
addition, regarding the high-quality hay diets, for each parameter, the CONTRAST
statement of SAS was used to determine the signicance of a linear or quadratic eect
of increasing amounts of high-quality hay. Because the treatments of high-quality hay
diets were unequally spaced (i.e. 60%, 75%, 100%), we used the ORPOL function
(PROC IML of SAS) to estimate the appropriate coecients for the linear and quadratic
CONTRAST statements.

3. Results
3.1. Dietary eects on intake and apparent nutrient digestibility
As expected by design, concentrate and hay intake diered signicantly among the
experimental groups (Table 2). There was a linear eect of dietary high-quality hay
proportion on total dry matter intake (DMI) (p < 0.05); however, no dierence
occurred between Groups 60HQ and 60NQ. NDF intake was also aected by the
experimental design, with a linear eect of dietary high-quality hay proportion in the
HQ hay diets (p < 0.001). Due to the high NDF content of the bre-rich hay, total NDF
intake of Group 60NQ was higher than in Groups 60HQ and 75HQ but was still lower
than in Group 100HQ (p < 0.001).
Table 2. Treatment eects on feed intake and apparent total tract digestibility (ATTD).
p-Values

Experimental diets*

Dry matter intake [kg/d]

Hay
Concentrate
Total
NDF intake [kg/d]
Hay NDF
Concentrate NDF
Total NDF
ATTD [%]
Dry matter
Organic matter
Crude protein
NDF
Acid detergent
bre
NFC
Ether extract

60NQ

60HQ

75HQ

100HQ

SEM

Diet

Linear
contrasts

7.74c
4.75a
12.5c

7.83c
4.92a
12.7bc

9.77b
3.16b
12.9ab

13.2a
0.00c
13.2a

0.144
0.087
0.204

<0.001
<0.001
0.001

<0.001
<0.001
0.017

0.169
0.014
0.625

6.10a
0.00d
6.08a

0.073
0.017
0.083

<0.001
<0.001
<0.001

<0.001
<0.001
<0.001

0.154
0.008
0.501

4.47b
0.928b
5.38b

3.64c
0.997a
4.63d

4.53b
0.647c
5.18c

Quadratic
contrasts

65.1b
67.8b
66.7b
59.6c
53.2c

73.6a
76.1a
72.1a
71.9b
67.0b

74.1a
76.5a
73.0a
77.1a
73.9a

73.5a
75.9a
75.5a
81.0a
78.1a

1.17
1.06
1.15
1.27
1.66

<0.001
<0.001
<0.001
<0.001
<0.001

0.915
0.786
0.009
<0.001
<0.001

0.607
0.608
0.665
0.088
0.089

79.7b
44.0a

84.1a
42.3a

80.5ab
38.8a

68.3c
20.0b

1.24
4.00

<0.001
0.003

<0.001
0.003

0.153
0.196

Notes: *60NQ, 60% normal hay + 40% concentrate; 60HQ, 60% high-quality hay + 40% concentrate; 75HQ, 75% highquality hay + 25% concentrate; 100HQ, 100% high-quality hay; SEM, standard error of the mean; linear and
quadratic contrasts were calculated for high-quality hay proportions in the high-quality hay containing diets; NDF,
neutral detergent bre; NFC, Non brous carbohydrates; a,b,cmeans in the same row with dierent superscripts dier
signicantly (p < 0.05).

M.-T. KLEEFISCH ET AL.

The ATTD of DM, OM, CP, NDF and ADF was lower in Group 60NQ than with the
high-quality hay groups (p < 0.001; Table 2). However, Group 100HQ showed with
20.0% and 68.3% the lowest digestibility of EE and NFC, respectively, compared to the
other groups (p < 0.001; Table 2). The ATTD of CP, NDF and ADF showed a linear
improvement with increasing proportion of high-quality hay in the diet (p < 0.001;
Table 2), whereas the ATTD of NFC (p < 0.001) and EE (p = 0.003) linearly decreased.

3.2. Dietary eects on time spent eating and ruminating

The time spent eating and ruminating diered depending on the diet type (Figure 1; Table 3).
The total daily time spent eating was reduced when the normal bre-rich hay was replaced by
high-quality hay (p < 0.001), whereas the time spent ruminating was longest in Group 100HQ
and shortest in Group 60HQ (p < 0.001). About 100 boli more per day were regurgitated in
Group 100HQ compared to the other groups (p < 0.001). Chews per bolus did not dier
between Groups 100HQ and 60NQ, but inclusion of concentrate in the HQ hay diets reduced
the chewing numbers per bolus (linear eect p < 0.001). Taken the chewing activity of eating
and ruminating together, the shortest time spent chewing was observed in Group 60HQ
where animals chewed approximately 125 min less than cows receiving diets 60NQ and
100HQ (p < 0.001).
A similar pattern was observed when calculating the time of eating and total chewing
per kg of DMI. The longest time spent eating was found in Group 60NQ (p < 0.001).
Time spent ruminating per kg DMI was signicantly shorter in Group 60HQ compared
with Groups 60NQ and 100HQ (p < 0.001) and there was a linear eect of the highquality hay proportion on time spent ruminating per kg DMI (p = 0.003).
Numbers of eating chews per kg DMI were higher in Group 60NQ compared to the
high-quality hay diets (p < 0.001). No dierences in numbers of ruminating chews per
kg DMI were found between Groups 60NQ and 100HQ, but numbers were lower when
concentrate replaced part of the high-quality hay (p = 0.001).

Figure 1. Ruminating activity over 24 h.

60NQ: 60% normal hay + 40% concentrate; 60HQ: 60% high-quality hay + 40% concentrate; 75HQ: 75% highquality hay + 25% concentrate; 100HQ: 100% high-quality hay. Dierences were found between diets
(p < 0.001) and time points (p < 0.001), but no diet time interaction occurred.

ARCHIVES OF ANIMAL NUTRITION

Table 3. Treatment eects on eating, ruminating and total chewing activity.

p-Values

Experimental diets*

Number of bolus [n/d]

Chews per bolus
Chews per min
Time [min/d]
Eating
Ruminating
Total chewing
Time [min/kg of DMI]
Eating
Ruminating
Total chewing
Chews [n/kg DMI]
Eating
Ruminating
Total chews
Time [min/kg of hay NDF# intake]
Eating
Ruminating
Total chewing
Chews [n/kg hay NDF intake]
Eating
Ruminating
Total chews
Time [min/kg of total NDF intake]
Eating
Ruminating
Total chewing
Chews [n/kg total neutral detergent bre
intake]
Eating
Ruminating
Total chewing

Linear
Quadratic
60NQ 60HQ 75HQ 100HQ SEM Diet contrasts contrasts
369b
348b
364b
470a 25.6 <0.001 <0.001
0.265
0.153
49.1a 39.4c 44.2b 48.6a 2.01 <0.001 <0.001
b
b
b
a
55.5
58.3
62.8
1.70 <0.001 <0.001
0.834
58.1
225a
380ab
605a

169b
311c
480b

183b
340bc
523ab

186b
420a
607a

17.9a
30.7a
48.6a

12.8b
23.7b
36.5b

13.7b
25.8ab
39.6b

13.7b
30.5a
44.2ab

1169a
1818a
2987a
49.8a
84.4a
134a

0.168
<0.001
0.002

0.408
0.621
0.949

0.001
0.004
0.002

0.347
0.003
0.013

0.311
0.930
0.665

804b
1354b
2157c

842b
833b 71.5 <0.001
1490b 1829a 129.2 <0.001
2333b 2661ab 184.3 <0.001

0.613
0.001
0.008

0.481
0.963
0.796

45.7ab
84.1a
130a

40.7b
74.4ab
115a

3.03 <0.001
5.59 0.005
8.00 <0.001

<0.001
0.004
<0.001

0.570
0.638
0.910

3258a 2836ab 2496b

5011a 4804ab 4320ab
8270a 7638ab 6817bc

1772c 192.5 <0.001

3943b 347.0 0.014
5715c 489.3 <0.001

<0.001
0.022
0.001

0.648
0.642
0.895

41.2a
70.8
112

29.2b
65.3
94.5

0.002
0.526
0.108

0.037
0.802
0.560

0.398
0.919
0.696

1779b 172.0 <0.001

3918 311.7 0.307
5697b 443.7 0.036

0.016
0.400
0.752

0.592
0.992
0.806

35.1ab
64.9
100

34.2ab
64.3
98.5

2693a 2205b 2093b

4193 3708 3709
6886a 5912ab 5805ab

29.1c
65.2b
94.3b

11.2 0.002
22.2 <0.001
30.1 0.002
1.11
2.10
3.00

2.67
5.06
7.22

Notes: *60NQ, 60% normal hay + 40% concentrate; 60HQ, 60% high-quality hay + 40% concentrate; 75HQ, 75% highquality hay + 25% concentrate; 100HQ, 100% high-quality hay; SEM, standard error of the mean; linear and
quadratic contrasts were calculated for high-quality hay proportions in the high-quality hay containing diets; sum of
eating and ruminating time; DMI, dry matter intake #NDF, neutral detergent bre; a,b,cmeans in the same row with
dierent superscripts dier signicantly (p < 0.05).

Relating the chewing activity to NDF intake from the forage source, animals in Group
100HQ spent 1020 min less eating per kg hay NDF intake than the other groups.
Numbers of eating and ruminating chews per kg hay NDF intake were lower in Group
100HQ (p < 0.001), whereas no dierence occurred between Groups 60HQ and 60NQ.
The eating time per kg of total NDF intake also was lower in Group 100HQ
compared with Group 60NQ (p = 0.001), with a linear decreasing eect of the highquality hay proportion (p < 0.05). More eating chews per kg total NDF were performed
in Group 60NQ compared to the high-quality hay diets (p < 0.001). The time and
numbers of chews spent ruminating per kg of total NDF intake, though, did not dier
between the experimental groups (p > 0.1; Table 3).
Regarding the temporal pattern of ruminating over the course of 24 h dierences
between diets became obvious (Figure 1; diet eect p < 0.001, time eect p < 0.001) but
no diet time interaction was signicant.

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M.-T. KLEEFISCH ET AL.

3.3. Dietary eects on pH of FRL and PARL

The measured pH-values in FRL and PARL depended both on the feeding regime
(p < 0.001) and the time of sampling (p < 0.001; Figure 2). Considering the mean pHvalues in FRL over all time points, cows receiving diets 60NQ and 100HQ had similar
ruminal pH of 6.85 and 6.83, respectively, both being signicantly higher than in the
other groups (p < 0.05). No dierence in FRL-pH was found between diets 60HQ and
75HQ (on average pH = 6.71). Regarding the sampling time points, pH-values in FRL
ranged from 7.19 as the highest values at 7:00 h in Group 60HQ to 6.15 in Group 60HQ
as the lowest value 10 h later. At 8 h after the morning feeding, the pH was signicantly
higher in animals fed diet 60NQ compared with diet 60HQ (6.8 vs. 6.4; p = 0.023).
However, at 10 h, no dierences between the concentrate containing diets in FRL-pH
appeared. In contrast, at 10 h, animals receiving diet 100HQ displayed with 6.7 the
signicantly highest FRL pH-value compared to all other experimental groups (p < 0.05;
Figure 2).
Regarding the average pH-values in PARL across all time points, animals in
Group 60NQ displayed the highest mean pH-value with 6.68 (p < 0.001), whereas
the lowest value of 6.21 was found in Group 60HQ. No dierences between groups
occurred at 0 h before the morning feeding and 2 h later. But 4 h after the morning
feeding, animals receiving any high-quality hay diet had a signicantly lower PARL
pH than Group 60NQ (p < 0.05), remaining like that until 8 h after the morning
feeding. At 10 h after the morning feeding, in Group 60NQ, the pH in PARL had
dropped to a level similar to values observed in Groups 60HQ and 75HQ, whereas
the pH in Group 100HQ increased again to 6.22 (signicant against Group 60HQ
and 75HQ, p < 0.001).

Figure 2. Dietary eect on the pH of the ventral free ruminal liquid (FRL) (panel a) and on the pH of
the particle associated liquid (PARL) in the rumen (panel b).
60NQ: 60% normal hay + 40% concentrate; 60HQ: 60% high-quality hay + 40% concentrate; 75HQ: 75% highquality hay + 25% concentrate; 100HQ: 100% high-quality hay; error bars indicate the standard error of the
mean.
Panel A: *60NQ vs. 60HQ: p < 0.05. 100HQ vs. others: p < 0.05; panel B: *60NQ vs. 60HQ: p < 0.05; **60NQ vs.
60HQ and 75HQ: p < 0.05; ***60NQ vs. others: p < 0.05; 100HQ vs. 60HQ and 75HQ: p < 0.001.

ARCHIVES OF ANIMAL NUTRITION

11

4. Discussion
4.1. Feed intake and apparent total tract nutrient digestibility
Using high-quality hay with elevated sugar content might be a feeding option to oer
enough protein as well as energy in form of sugars and at the same time to provide
enough brous structure to maintain a sucient chewing activity and thus saliva
production to stabilise ruminal pH. Fibre content is known to inuence DMI (Allen
1996); however, 46% of NDF in Diet 100HQ might still be comparable to the NDF
content in Diet 60NQ (42%) and no depression in DMI occurred. On the contrary,
DMI slightly improved linearly with an increasing proportion of high-quality hay. A
possible reason might be a high palatability of the high-quality hay in combination with
a high digestibility (Lee et al. 2002; Moorby et al. 2006). However, it needs to be
considered that in this study, the overall DMI was low because of the use of nonlactating cows.
Usually, the accumulation of sugars in forages is associated with a reduction of the
bre structural carbohydrates (Burns et al. 2007), thus improving DM digestibility
(Hoover and Stokes 1991), as it was the case in the present experiment. Water-soluble
carbohydrates, like sugars, are rapidly available for ruminal microbes and are degraded
faster (Hoover and Stokes 1991; Oba 2011). Our results are in agreement with Miller
et al. (2001) who also found a higher DM and bre digestibility with grasses high in
sugars compared to a control grass. Although NDF digestibility was improved in Group
60HQ compared to Group 60NQ, it was still considerably lower in comparison to
Group 100HQ. Two possible explanations exist. On the one hand, the NDF of the
concentrate might be less digestible than the NDF from the high-quality hay. On the
other hand, the lower ruminal pH found in Groups 60HQ and 75HQ 10 h after the
morning feeding could have adversely aected bre degradation. Also diets rich in
concentrate seem to favour the growth of amylolytic bacteria, whereas numbers of
cellulolytic bacteria have been shown to decrease (Fernando et al. 2010), thus lowering
the degree of bre degradation.
The ATTD of NFC and EE was lowest in cows eating high-quality hay only, possibly
because of their forage origin. The NFC content of the 100HQ diet was in general low
and NFC in forages are not composed of easily degradable starch as it is the case in
grains. However, sugars, which constituted the largest proportion of NFC in the highquality hay, were thought to be completely degraded (Hoover and Stokes 1991; Oba
2011), thus the lower NFC digestibility in Group 100HQ was unexpected. It might be
that part of the sugars was trapped in lignied bre and thus could not be targeted by
enzymes or get absorbed in the small intestine. Ether extracts from forages and grains
dier considerably in their constituents, with forages containing less triglycerides than
grains and seeds, but more nonsaponiable substances like waxes, chlorophyll and cutin
(Palmquist and Jenkins 2003). In contrast to nonsaponiable lipids, triglycerides are
considered to be completely metabolisable (Palmquist and Jenkins 2003).
4.2. Structural eectiveness of high-quality hay and the eects on ruminal pH
Traditionally, chewing and ruminating activities are considered important noninvasive
assessable parameters of structural eectiveness and thus ruminant health because both

12

M.-T. KLEEFISCH ET AL.

parameters are directly aected by cell-wall content (Welch and Smith 1969) and
particle size (Tafaj et al. 2005). However, as in the present study, the particle sizes of
Diets 60NQ and 60HQ (both hays had approximately 1012 cm length, the concentrate
was oered as meal) were similar, the diering responses of chewing activity among
these diets can mainly be attributed to other structural properties such as diering
contents of forage NDF. Welch and Smith (1969) reported that bre is the nutritional
component of forages that is related to chewing activity, usually reported in the
literature as minutes per kg of NDF intake. Accordingly and in agreement with others
(Beauchemin 1991; Armentano and Pereira 1997), the total time spent ruminating per
day increased with increasing NDF concentration in the diet revealing a similar
ruminating activity of animals fed the diets 60NQ and 100HQ either in min/d or
min/kg DMI, indicating a similar structural eectiveness of the diets. When part of
the high-quality hay was replaced by concentrate, however, the dietary NDF content
dropped and ruminating activity decreased signicantly, indicating a loss in structural
eectiveness. This is in agreement with other studies that found decreased chewing
activity and chewing activity per kg DMI when the forage-to-concentrate ratio
decreased (Sudweeks et al. 1975; Sudweeks 1977; Yang and Beauchemin 2009).
Although cows of Group 100HQ spent the shortest time ruminating and chewing per
kg hay NDF, no dierences between diets were found in time ruminating per kg total
NDF, supporting the results that concentrate as part of the other diets decreased the
overall time spent ruminating and chewing.
Interestingly, eating time related to hay NDF intake and numbers of chews per kg
hay NDF intake were shortest and lowest in Group 100HQ, possibly as a result of an
improved chewing eciency in response to feeding a forage-only diet. As reviewed by
De Boever (1990), there are several factors causing a more ecient chewing like e.g. a
greater jaw force (Hooper and Welch 1983) or a higher number of chews per time (Bae
et al. 1981), as observed in Group 100HQ. As demonstrated by Bae et al. (1981), cows
seem to be able to consume more hay daily by increasing chewing time and the speed of
chewing. However, contrasting to their ndings, we did not observe increased eating
time with increased amount of hay intake. Apparently, the dietary proportion of forage
is not the only parameter aecting feed intake but also the bre quality is of inuence
too. Eating time and number of eating chews were always longest and highest with diet
60NQ supporting the fact that the bre fraction of diet 60NQ was more lignied than
the NDF of the high-quality hay diets and needed more time for an ecient mastication
and cell wall breakdown, despite of the 40% concentrate in the ration.
An adequate chewing activity and secretion of alkaline saliva are crucial for neutralisation of ruminal fermentation acids to stabilise ruminal pH (Allen 1997; Mertens
1997). Investigations on the eect of hay with elevated sugar content on ruminal pH are
scarce, although it was shown in several studies that sugar supplementation in form of
mono- or disaccharides aects ruminal pH and can even increase it (reviewed by Oba
2011). Few studies investigated the eects of fresh grass with high sugar content on
ruminal pH (Lee et al. 2002; Taweel et al. 2005). For example, no eect on ruminal pH
was observed by Taweel et al. (2005) who oered high-sugar grass and concentrate in
the proportion 80:20 on DM basis to dairy cows. However, in general, they observed a
rather low pH of 5.86.1 and Lee et al. (2002) found a slightly increased pH in sheep
when oered high-sugar grass but they did not supply additional concentrate. As

ARCHIVES OF ANIMAL NUTRITION

13

expected, in the present study, the pH decreased with time after feeding. However, in
the time course of 10 h, only two animals experienced a FRL pH below 5.8, the value
considered as threshold for subacute ruminal acidosis SARA (Zebeli, Dijkstra, et al.
2008). These animals were fed diets 60HQ and 75HQ, respectively, and the low pHvalues were measured 8 h after the rst morning feeding. It might have been expected
that the lowest pH values would be observed in cows on a concentrate diet in
combination with the high-quality hay, as starch from concentrate and sugars from
forage are easily fermentable carbohydrates, although pH enhancing eects of sugars
have been reported too (Oba 2011; Gao and Oba 2016). The fast fermentation of starch
and sugars will lead to a quick production of volatile fatty acids (VFA) possibly causing
a decrease in ruminal pH. However, unlike starch, the addition of soluble NDF in form
of beet pulp has even been reported to have ruminal pH stabilising or enhancing eects
(Mahjoubi et al. 2009; Mojtahedi and Danesh Mesgaran 2011) and thus is not considered as attributable factor to the observed lower pH in Groups 60HQ and 75HQ in
this study. Changes in pH became especially obvious when measured in PARL. This
PARL deriving from the ruminal mat generally shows higher VFA concentrations and
lower pH than FRL (Tafaj et al. 2004; Klevenhusen et al. 2014). A low pH has been
shown to inhibit brolytic bacteria (Russel and Wilson 1996; Granja-Salcedo et al. 2016)
and thus bre degradation, which largely occurs in the bre mat (Tafaj et al. 2004).
Accordingly, the lower pH observed in PARL supports the results of the lower bre
degradability observed in the grain containing diets (linear eect of high-quality hay
proportion p < 0.001). The observed lower pH in Groups 60HQ and 75HQ can also be
supported by the chewing results, as less time per day was spent chewing and ruminating in those groups due to the decreased structural eectiveness of the diets.
In Group 60NQ, ruminal pH remained relatively stable, especially in PARL, despite
of the 40% concentrate in the diet and dropped later than with the high-quality hay
diets. This pH-prole and the digestibility results again indicate that the brous
carbohydrates from the normal hay were harder to digest than from the high-quality
hay and might need longer time to be fermented. Thus, a further drop in pH after 10 h
might be expected but has to be proven with indwelling pH sensors to reveal diurnal
changes. Although ruminal pH dropped in Group 100HQ with time after the rst
morning feeding similarly to the other treatments, it again increased considerably at
10 h after the morning feeding (6.7 in FRL and 6.2 in PARL). In contrast, the ruminal
pH values from all concentrate containing treatment groups were still below 6.4 in FRL
and below 5.9 in PARL at 10 h. Apparently, the structural eectiveness of the highquality hay was sucient to induce enough chewing and ruminating activity and with
them saliva production to neutralise the fermentation acids in contrast to the concentrate and thus starch containing diets.

5. Conclusions
Results suggest benecial eects of replacing common bre-rich hay and concentrate
with high-quality hay with elevated sugar content on OM, bre and protein digestibility. Although overall eating time was considerably reduced when high-quality hay
was oered, feeding 100% high-quality hay stimulated chewing and ruminating activities and improved chewing eciency, thus stabilising ruminal pH. Adding starch-rich

14

M.-T. KLEEFISCH ET AL.

concentrate to the high-quality hay diet, though, lowered the structural eectiveness of
the diet and resulted in diminished ruminating behaviour and decreased ruminal pH.
Further research with lactating dairy cows is needed to determine whether feeding highquality hay might be an alternative feeding strategy as opposed to concentrate feeding
to improve ruminal health without impairing metabolic health status and milk production parameters.

Acknowledgements
The authors are grateful to Dr W. Pohl, Dr J. Huber, E. Draxler, A. Kfer and the sta of the
dairy research station Kremesberg at the University of Veterinary Medicine Vienna for their
excellent collaboration. The skilful assistance of Anita Dockner, Melanie Wild, Sandra Eisen and
Sabine Leiner (Institute of Animal Nutrition and Functional Plant Compounds, University of
Veterinary Medicine, Vienna) with nutrient and TiO2 analyses is very much appreciated. The
high-quality hay was kindly provided by ARGE Heumilch Austria.

Disclosure statement
No potential conict of interest was reported by the authors.

Funding
This work was supported by The Austrian Federal Ministry of Agriculture, Forestry,
Environment and Water Management [Grant Number BMLFUW100928].

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