Livestock Science 191 (2016) 22–28

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Livestock Science
journal homepage: www.elsevier.com/locate/livsci

Variation of TMR particle size and physical characteristics in

commercial Iranian Holstein dairies and effects on eating behaviour,
chewing activity, and milk production
M. Esmaeili a, M. Khorvash a, G.R. Ghorbani a, S.M. Nasrollahi b,n, M. Saebi a
a
Department of Animal Sciences, Isfahan University of Technology, Isfahan 84156-83111, Iran
b
Young Researchers Club, Khorasgan (Isfahan) Branch, Islamic Azad University, Isfahan, Iran

art ic l e i nf o a b s t r a c t

Article history: Evidence is limited on the effect of physical form of total mixed rations (TMR) on management of group-
Received 19 October 2015 housed dairy cows in commercial dairy herds. The objective of this study was to consider the variation of
Received in revised form TMR particle size (PS) and physical characteristics in commercial dairies and its effects on eating be-
14 June 2016
haviour, chewing activity, and milk production. Fifteen dairy herds, feeding TMR, were each studied for
Accepted 4 July 2016
7 consecutive days. In each herd, a group of multiparous (3.57 0.8 parity; mean7 SD), high producing
cows (47 74 kg milk/d) in mid-lactation (1097 12 DIM) was studied. Herds were visited for 7 con-
Keywords: secutive days for sampling fresh and refusal feed from the test group to assess particle size distribution,
Particle size dry matter, and chemical composition. Chewing activity and milk production were monitored for the
TMR
group. The 15 herds were divided to 3 classes according to geometric mean particle size of TMR: short,
Feeding behaviour
medium, and coarse, with average lengths of 4.6, 5.4, and 6.7 mm respectively. The classes were sig-
Milk production
Dairy farm nificantly different in geometric mean particle length (GMPL) and physically effective neutral detergent
fibre1.18 (peNDF1.18) (P o0.01), but physically effective neutral detergent fibre8 (peNDF8) was similar
between medium and coarse classes. Amount of TMR retained on the 19-mm sieve of Penn State Particle
Separator was highest for dairy farms with medium GMPL (Po 0.01). Diurnal changes in particle size of
refusals and chewing activity were similar among classes; the only difference observed was that daily
rumination activity in the medium class tended to be greater than for the short and coarse classes
(P ¼0.10). Milk fat percentage and yield as well as fat-to-protein ratio (F:P) were higher in the classes
with medium and coarse TMR PS (Po 0.01). Although GMPL and peNDF1.18 of TMR were correlated
(P o0.05) only with milk F:P (r ¼0.53 and 0.64, respectively), peNDF8 was correlated (Po 0.05) with both
milk F:P (r ¼ 0.56) and chewing activity (r ¼0.60). The 19-mm TMR fraction was the only variable that
correlated (Po 0.05) with milk fat percentage (r¼0.51) along with a tendency to be correlated (P¼ 0.09)
with chewing time. Interestingly, grain percentage of TMR was negatively correlated (P o 0.05) with both
milk F:P (r ¼  0.58) and chewing time (r ¼  0.68). In conclusion, commercial dairy farms considered in
this study had a high variation in TMR PS, and peNDF8 and the 19-mm fraction of the Penn State Particle
Separator were good indicators for predicting bio-physical responses of cows.
& 2016 Elsevier B.V. All rights reserved.

1. Introduction
Physical form of the diet is a double-edged sword when con-
sidering dairy cow profitability. Diets with fine particle size (PS)
and low NDF may induce acidosis, milk fat depression, laminitis,
and displaced abomasum; diets with extra coarse particles induce
sorting, fill the rumen, and reduce DMI and milk production
n
Corresponding author.
E-mail addresses: smnasrolahi@gmail.com,
smnasrolahi@ut.ac.ir (S.M. Nasrollahi).
(Heinrichs and Kononoff, 2002; Cherney and Cherney, 2003, Nas-
rollahi and Khorvash, 2014). Physically effective NDF (peNDF) was
developed to adjust the chemical value of fiber (NDF) for physical
form (i.e., particle size) (Mertens, 1997). Numerous studies de-
scribe the effect of peNDF of the diet on dairy cow health and
profitability (Mertens, 1997; Teimouri Yansari et al., 2004; Zebeli
et al., 2012). Interaction with forage source (Bhandari et al., 2007,
2008), grain fermentability (Krause et al., 2002a, 2002b; Nasrollahi
et al., 2012, 2014), and NDF source (Alamouti et al., 2009) have
been previously documented along with some recommendations
(Mertens,1997; Zebeli et al., 2008).

http://dx.doi.org/10.1016/j.livsci.2016.07.003
1871-1413/& 2016 Elsevier B.V. All rights reserved.

Please cite this article as: Esmaeili, M., et al., Variation of TMR particle size and physical characteristics in commercial Iranian Holstein
dairies and effects on eating behaviour, chewing activity, and milk production. Livestock Science (2016), http://dx.doi.org/10.1016/j.
livsci.2016.07.003i
 M. Esmaeili et al. / Livestock Science 191 (2016) 22–28 23

Factors reducing forage and TMR PS in commercial settings are Table 1

many, including harvesting, storing, removing from storage, Ingredients and chemical characteristics of TMRa, and the DM and pH of corn silage
fed dairy farms in the study (n¼ 15).
chopping, and mixing (Heinrichs, et al., 1999; Nasrollahi and
Khorvash, 2014). The geometric mean particle length (GMPL) and Variable Mean Minimum Maximum SD CV %
distribution of TMR particles are influenced by these factors, along
with botanical characteristics of forages and climate condition TMR ingredients % on DM basis
(such as temperature, humidity, and rainfall). Accordingly, final PS Lucerne hay 15.7 8.0 23.8 3.96 25.28
Maize silage 19.9 15.5 24.6 2.28 11.47
of TMR fed to dairy cows might vary due to the equipment and Concentrate 64.4 58.0 71.0 3.42 9.60
time used at each step of PS reduction, as well as different forage Beet pulp 6.2 3.5 9.1 1.53 24.85
sources, maturity, and climatic conditions. Inadequate mean PS of Grain 34.6 26.0 44.4 4.18 12.07
TMR, as well as exposure to an inconsistent diet, can lead to de- Ground barley 16.4 0 29.4 8.91 54.31
Ground corn 18.2 5.9 35.1 8.7 47.70
pressed rumen pH and an increased risk of ruminal acidosis
TMR chemical analysis
(DeVries et al., 2008; Zebeli et al., 2012). Understanding the opti- DM, % 47.1 37.3 55.0 4.13 8.77
mum TMR PS allows producers to manage factors affecting TMR PS CP, % of DM 17.0 15.5 18.4 0.85 5.00
that they can control. EE, % of DM 4.5 3.5 5.4 0.64 14.19
The majority of research into ration physical form has been in Ash, % of DM 7.5 5. 9 8.5 0.78 10.36
NDF, % of DM 30.2 27.2 33.0 1.67 5.52
controlled experiments (Nasrollahi et al., 2015). Though this re- ADF, % of DM 18.0 16.4 20.6 1.27 7.06
search has been central to understanding the relationship between NFCb, % of DM 40.7 37.7 43.8 1.84 4.52
feed PS and cow health and performance, there is limited data on Silage parameters
the effect of TMR PS at the herd level under commercial condi- pH 4.10 3.87 4.39 0.15 3.67
DM, % 22.1 19.3 26.9 1.96 8.90
tions. Indeed, particular effects of group feeding, such as compe-
tition at the feed bunk and sorting particles between dominant a
National Research Council (NRC, 2001), Cornell Net Carbohydrate and Protein
and subordinate cows (Hosseinkhani et al., 2008), herd-to-herd System (CNCPS) version 5.0 (Fox et al., 2000), and Cornell-Penn-Miner Dairy (CPM-
and day-to-day variation in nutrient composition of TMR (Sova Dairy) Version 3.08.01 (http://www.cpmdairy.com/In-dex.html) were the systems
et al., 2014), as well as stressors, such as heat and cold, waiting used for ration formulation among dairy farms.
b
NFC ¼ 100  [CP þ(NDF  NDICP) þfat þash].
time to milking, and the distance between the resting area and
bunk, may influence dairy cow responses to PS in commercial
settings. It may not be possible to consider all these variables in a animals with high milk production (47.3 73.69 kg/d).
controlled experiment. Leonardi and Armentano (2007) compared
group and individual feeding of dairy cows and reported cows in 2.3. Housing and feeding management
group feeding situations were more prone to sort feed due to
competition. Cows were housed in the farms' normal grouping systems, with
To our knowledge there are limited studies considering the no feeding and management changes through the study period
variation in TMR PS across commercial dairy farms. Therefore the (Table 1). Cows on all farms were housed in freestalls with a ratio
objectives of the study were 1) to consider the variation in TMR PS of 1.25 stalls/cow. The groups were stable throughout the study
and other physical characteristics as well as fermentability on period with no cows added or removed. Cows were fed 3–4 times
commercial dairy farms and 2) to consider the relationship be- daily and rations were prepared in feed mixer wagons of 6,000 kg
tween TMR physical characteristics and cow response in com- capacity with 3 horizontal augers. Mixing time averaged 15 min/
mercial settings. We hypothesized that variation among Iranian batch, depending on manual control of PS and the time required
dairy herds in TMR PS would affect eating behaviour, chewing, and for delivering feed throughout the length of the bunk. During
milk component production of dairy cows. chopping and mixing, lucerne hay was followed by concentrate
mix, maize silage, beet pulp (tempered;  25% DM), and then
water. The amount of water added was different on each farm and
2. Materials and methods was based solely on a management decision by each dairy. Barley
and corn were used as grain sources and ground using hammer
2.1. Location mills with 2–3-mm screen size.

The experiment was conducted in Isfahan, Iran, between Jan-
uary and April 2012. The area is located at 32°39′N and 51°41′E at
an altitude of 1,600 m. Annual mean rainfall is 200 mm, and the
climate is dry and hot. During the experiment, average daily
temperatures were between 4.6 and 14 °C, and rainfall was be-
tween 3.5 and 34.8 mm per month.
2.2. Farm and cow selection
Fifteen commercial dairy farms were selected from 200 herds
according to size (biggest) and management criteria such as
feeding TMR, housing in freestall barns, and milking with an au-
tomated system. Animals were cared for according to the guide-
lines of the Iranian Council of Animal Care (1995). This study was
conducted in a manner that avoided unnecessary discomfort to the
animals. All animals were under the care of a veterinarian. For
each farm, a group of cows (83 738) was subjected to sampling
and data collection for 7 d. The cows in selected groups were
multiparous (parity of 3.54 70.79), mid-lactation (109 712 DIM)
2.4. Particle size measurement, average DMI, and chemical analysis
Samples of fresh feed were taken at the time of feeding for 6 d,
and refusals were sampled at 1, 2, 3, and 4 h post feeding for 2 d.
Duplicate samples were collected at these times to assess particle
size and the DM and chemical content of the ration. Fresh feed was
sampled from 5 different areas of the feed bunk (100 g/area), at
the beginning, middle, and end of TMR delivery, and within
minutes of delivery to ensure accurate representation of the TMR.
All TMR samples were composited and a final 2 kg sample was
obtained for each day. Refusals were sampled subsequent to bunk
clean-up and were mixed to obtain a representative sample (2 kg).
All samples were frozen at  10 °C until processing. Average DMI
was calculated by the daily difference between the amount of TMR
fed and refusals based on DM divided by the number of cows in
each group.
Samples for PS separation were thawed and separated using
the 3-screen (19, 8, and 1.18 mm) Penn State Particle Separator
(PSPS; Kononoff, 2002) with a horizontal sieving frequency of

Please cite this article as: Esmaeili, M., et al., Variation of TMR particle size and physical characteristics in commercial Iranian Holstein
dairies and effects on eating behaviour, chewing activity, and milk production. Livestock Science (2016), http://dx.doi.org/10.1016/j.
livsci.2016.07.003i
24 M. Esmaeili et al. / Livestock Science 191 (2016) 22–28

1.1 Hz or approximately 1.1 shake per second. The PSPS separated Table 2
the particles into 4 fractions: long ( 419 mm), medium ( o19 and Herd-to-herd variation in geometric mean particle length (GMPLa) of TMR fed on
15 commercial dairy farms (n ¼15).
Z8 mm), short (o 8 and Z1.18 mm), and fine (o1.18 mm) par-
ticles. After separation, the DM of each separated fraction was Variable Mean Minimum Maximum SD CV %
determined by oven-drying at 55 °C for 48 h. Particle size dis-
tribution (%) was calculated on a DM basis by dividing each dried GMPL, mm 5.34 4.12 6.64 0.78 14.59
fraction weight by the total dried sample weight. Physically ef- % of DM retained on screen
19 mm 8.23 3.60 14.80 3.57 43.41
fective NDF is calculated as dietary NDF content (%DM) multiplied 8 mm 32.52 24.90 39.60 4.69 14.42
by physical effectiveness factor determined as the DM proportion 1.18 mm 45.16 39.00 57.60 5.86 12.99
of particles retained on 2 sieves (19 and 8 mm; pef 8) or 3 sieves Pan 13.71 2.30 21.90 6.66 48.55
(19, 8, 1.18 mm; pef 1.18), respectively, of the PSPS (Lammers et al., peNDF8b 12.43 10.23 15.89 1.66 13.38
peNDF1.18b 26.06 22.84 30.63 2.45 9.42
1996; Kononoff, 2002). Samples taken for DM and chemical ana-
lysis were oven-dried at 55 °C for 48 h and then ground to pass a
Geometric mean particle length as calculated by ASAE (1995; method S424.1).
through a 1-mm screen (Brinkmann Mill, Brinkmann Instruments b
peNDF8, peNDF1.18¼ physically effective NDF calculated as dietary NDF
Co., Westbury, NY). Dried TMR samples were analyzed for CP using content (% DM) multiplied by physical effectiveness factor determined as the DM
the Kjeldahl method (Kjeltec 1030 Auto Analyzer, Tecator, Höga- proportion of particles retained on 2 sieves (19 and 8 mm; pef8) or 3 sieves (19, 8,
1.18 mm; pef1.18), respectively, of the PSPS (Lammers et al., 1996; Kononoff, 2002).
näs, Sweden; AOAC, 2002, method 955.04), ether extract (EE;
AOAC, 2002, method 920.39), ash (AOAC, 2002; method 942.05),
and NDF using heat stable α-amylase (100 μL/0.5 g of sample) and Table 3
Chemical composition and physical characteristics of TMR fed on dairy farms with
sodium sulfite (Van Soest et al., 1991). Neutral detergent insoluble different TMR particle size (three classes of short, medium, and coarse; n ¼ 5).
crude protein (NDICP) was measured by analyzing the content of
CP in NDF. Non-fiber carbohydrate (NFC) was calculated as 100  Variable TMR PS class P value
[CP þ(NDF  NDICP) þEE þash].
Short Medium Coarse

2.5. Chewing activity TMR chemical analysis

DM, % 49.17 1.57 48.67 1.57 43.7 7 1.57 0.06
Chewing activity was monitored visually for all cows in each CP, % of DM 17.3 7 0.40 16.9 7 0.40 16.9 70.40 0.79
EE, % of DM 4.69 7 0.30 4.477 0.30 4.42 7 0.30 0.80
group over a 24-h period except for milking time and the time
Ash, % of DM 7.4 7 0.37 7.5 7 0.37 7.7 7 0.37 0.83
cows were waiting for milking. Eating and ruminating activities NDF, % of DM 29.5 7 0.69 31.4 7 0.69 29.7 7 0.69 0.16
were noted by an observer at 5-min intervals, and each activity ADF, % of DM 17.3 7 0.54 18.7 70.54 17.9 7 0.54 0.24
1
was assumed to persist for the entire 5-min interval (Colenbrander NFC , % of DM 41.2 7 0.82 39.8 7 0.82 41.2 7 0.82 0.39
et al., 1991). At each visit, the number of cows with each activity GMPL2, mm 4.55 7 0.11a 5.36 7 0.11b 6.08 7 0.10c o0.01
% of DM retained on screen
were noted and the percentage of cows for each activity was cal- 19 mm 6.63 7 0.60a 10.38 7 0.63b 7.08 7 0.58a o0.01
culated (number of cows with each activity/total number of cows 8 mm 31.377 0.85a 30.92 7 0.89a 35.6970.83b o0.01
in group, multiplied by 100). At the end of 24-h, total time used for 1.18 mm 42.157 0.93a 43.86 7 0.97a 50.03 7 0.91b o0.01
each activity was calculated by multiplying the average percentage Pan 19.84 7 0.81c 14.83 7 0.84b 6.487 0.79a o0.01
peNDF83 11.43 7 0.27a 12.74 70.28b 12.99 7 0.26b o0.01
of each activity by the total time of measuring (24 h).
peNDF1.183 24.07 70.31a 26.25 7 0.32b 27.78 70.30c o0.01

2.6. Milk production and body condition score a–c

In each group 20 cows with milk production closest to the
group mean (according to previous herd records) were selected
and monitored for milk production. The number of selected cows
was constant across the farms and was based on the availability of
recording and sampling. Cows were milked 3–4 times daily in a
milking parlor. Milk yield was recorded, and samples were col-
lected into vials, preserved at 4 °C, and analyzed within 24 h for
fat, true protein, and lactose content (134 BN Foss Electric,
Hillerød, Denmark). Body condition score (BCS) for each cow in
each group was assessed by 2 observers according the method
described by Ferguson et al. (1994).
2.7. Statistical analysis
Prior to analyses, data outcomes were screened for normality
using the UNIVARIATE procedure of SAS (SAS Institute, 2002).
According to measured GMPL, the 15 dairy farms were equally
(n ¼5) divided into 3 classes (coarse, medium, and short) and
further analyses were made accordingly.
Geometric mean particle length, peNDF8, peNDF1.18, and dis-
tribution of PS were analyzed in a model that included diet PS as a
fixed effect and forage-to-concentrate ratio as a covariate. The
general linear models procedure of SAS (SAS Institute, 2002) was
used. The forage-to-concentrate ratio was used as a covariate to
adjust the effect of PS on physical characteristics of TMR. It re-
mained in the model for all physical characteristics variables
Means within a row with different superscripts differ (P o 0.05).
1
NFC¼ 100  [CP þ(NDF  NDICP) þ fatþ ash].
2
Geometric mean particle length as calculated by ASAE (1995; method
S424.1).
3
peNDF8, peNDF1.18¼ physically effective NDF calculated as dietary NDF
content (% DM) multiplied by physical effectiveness factor determined as the DM
proportion of particles retained on 2 sieves (19 and 8 mm; pef8) or 3 sieves (19, 8,
1.18 mm; pef1.18), respectively, of the PSPS (Lammers et al., 1996; Kononoff, 2002).
reported in Table 3.
Data on refused particles retained on PSPS sieves across time
and chewing activity were analyzed using the mixed procedure of
SAS (SAS Institute, 2002), with a fixed effect of forage PS and a
random effect of dairy farm, and forage percentage in the ration as
a covariate. Repeated measurements of the change in particles
retained on PSPS sieves across time were analyzed by including a
REPEATED statement, as well as terms for time and interaction of
treatment by time.
Milk production and composition data were analyzed using the
mixed procedure of SAS (SAS Institute, 2002) with a fixed effect of
forage PS and random effect of dairy farm. Average DMI, parity
number, BCS and grain percentage were considered as covariate
(when they were significant). BCS and grain percentage were used
as covariables to adjust the real effect of PS on milk production.
Pearson correlation coefficients were used to evaluate re-
lationships between the TMR physical characteristics and dairy
cow responses (group mean). For all models, all values were LSM
and effects were considered significant at Pr0.05 and tendencies
at Pr0.10.

Please cite this article as: Esmaeili, M., et al., Variation of TMR particle size and physical characteristics in commercial Iranian Holstein
dairies and effects on eating behaviour, chewing activity, and milk production. Livestock Science (2016), http://dx.doi.org/10.1016/j.
livsci.2016.07.003i
 M. Esmaeili et al. / Livestock Science 191 (2016) 22–28 25

3. Results Table 4
Changes in geometric mean particle length over time (h relative to feeding) and
peNDF in diets of dairy farms divided into short, medium, and coarse TMR particle
3.1. Ration and silage characteristics
size classes (n ¼5).

Ration ingredients and chemical composition of diets fed by Variable TMR PS class P value
each dairy farm are shown in Table 1. Lucerne hay and maize si-
lage were the 2 forage sources used in the diets and constituted Short Medium Coarse

about 36% of diets DM. The amount of lucerne hay used was more GMPL , mm1

variable (CV ¼25.3 vs. 11.5%) and lower on average (15.7 vs. 19.9%) 0h 4.59 7 0.24a 5.317 0.23b 6.23 7 0.23c 0.001
than maize silage. Beet pulp and grain were used on all farms, with 1h 5.08 7 0. 311a 6.067 0.310b 6.80 7 0.309b o0.01
inclusion at about 6% and 35%, respectively. The amount of grain 2h 5.27 70.420a 6.23 7 0.418a 7.277 0.417b 0.02
3h 5.60 7 0.514a 6.78 7 0.512a,b 7.577 0.519b 0.06
used had a relatively high variation (CV ¼ 12.1%) with a minimum
4h 6.65 7 0.880a 7.79 7 0.878a,b 9.38 7 0.875b 0.11
of 26 and maximum of 44%. Variation in chemical composition peNDF82
(CV ¼4.5 to 14.2%) was lower than variation in ingredients 0h 11.077 0.65a 13.08 70.63b 13.54 7 0.62b 0.04
(CV ¼9.6 to 54.3%). Diet DM averaged 47% with a wide range 37– 1h 12.007 0.88a 14.60 70.79b 14.26 7 0.78b 0.10
55%; CV ¼8.8%). The proportion of EE and ash in the diet was more 2h 12.84 7 1.05 15.22 71.03 14.83 7 1.03 0.26
3h 13.26 7 1.14 16.28 71.13 15.50 7 1.12 0.19
varied than other nutrients. The pH and DM of silage were rela- 4h 14.88 7 1.48 19.377 1.47 17.53 71.46 0.13
tively consistent across dairy farms with CV of 3.7% and 8.9%, re- peNDF1.182
spectively. These results suggest that conditions for ensiling corn 0h 23.86 7 0.74a 26.26 7 0.73b 27.73 70.72b 0.008
silage were relatively similar across farms. 1h 24.387 0.65a 26.80 7 0.64b 28.25 7 0.63b o0.01
2h 24.527 0.68a 26.87 7 0.68b 28.34 7 0.68b o0.01
3h 24.72 70.65a 27.137 0.65b 28.40 7 0.65b o0.01
3.2. Particle length and physically effective fiber 4h 25.20 7 0.66a 28.187 0.66b 28.74 70.65b o0.01

a–c
There were high variabilities among TMR physical form and CV Means within a row with different superscripts differ (P o 0.05).
1
was more than 10% in almost all items (Table 2). The greatest Geometric mean particle length as calculated by ASAE (1995; method S424.1)
with a sieving frequency of 1.1 Hz or approximately 1.1 shake per second.
variabilities were observed for longer particles (419 mm) and 2
peNDF8, peNDF1.18¼ physically effective NDF calculated as dietary NDF
material retained on the pan ( o1.18 mm). The GMPL and peNDF8 content (% DM) multiplied by physical effectiveness factor determined as the DM
were more variable than peNDF1.18, which had the lowest varia- proportion of particles retained on 2 sieves (19 and 8 mm; pef8) or 3 sieves (19, 8,
tion (CV ¼9.4%). 1.18 mm; pef1.18), respectively, of the PSPS (Lammers et al., 1996; Kononoff, 2002).
As intended, TMR on dairy farms in the 3 classes were similar
in chemical composition, but different in physical characteristics Table 5
(Table 3). Means of GMPL were 4.6, 5.4, and 6.1 mm for short, Chewing behaviour on dairy farms divided in short, medium, and coarse TMR
particle size classes (n¼ 5).
medium, and long classes, respectively (Po 0.01). The amount of
particles 4 19 mm was highest on dairy farms with medium Variable TMR PS class (mean7 SEM) P value
GMPL, but particles retained on the 8-mm and 1.18-mm sieves
were greater for the coarse class (Po0.01). The particles retained Short Medium Coarse
on the bottom pan of the PSPS were significantly decreased with
%
increasing GMPL in the TMR. Amount of peNDF8 was similar be- Eating 23.4 7 0.89 23.1 71.02 24.37 0.85 0.61
tween the medium and coarse classes and was more than the Rumination 32.4 7 0.61 34.8 7 0.68 33.77 0.61 0.07
short class (Po 0.01). In contrast, peNDF1.18 followed the same Total chewing 55.8 7 1.17 58.0 7 1.34 58.0 7 1.11 0.35
pattern as GMPL and was 24.1%, 26.3% and 27.8% for short, medium Min day  1
Eating 337 714.1 3337 16.8 3507 14.1 0.61
and long classes, respectively (P o0.01).
Rumination 4677 8.8 501 79.8 486 7 8.8 0.07
Total chewing 803 718.8 8357 21.4 836 7 17.9 0.35
3.3. Change in particle characteristics and chewing activity over time

Data on the change in GMPL, peNDF8, and peNDF1.18 during Table 6

the first 4 h after morning feeding are shown in Table 4. Regardless Milk production and composition on dairy farms divided into short, medium, and
coarse TMR particle size classes.
of treatment, the GMPL and peNDF in the refusal increased as time
increased (P o0.05). Although the 3 classes of short, medium, and Variable TMR PS class (mean 7 SEM) P value
coarse were different in GMPL of TMR at the beginning of the day,
these differences were not consistent during the day. At 1 h post Short (n¼ 97) Medium Coarse
(n ¼ 100) (n¼ 100)
feeding, the classes of medium and coarse had similar GMPL, and
at 2 h short and medium were similar. The condition of peNDF8 in Milk (kg/d) 41.8 7 2.26 46.17 2.29 44.4 7 2.26 0.30
refusal at various times after feeding was rather different relative 3.5% FCM (kg/ 31.7 7 0.90 33.57 0.92 32.8 7 0.91 0.30
to GMPL. Although farms with short PS had lower peNDF8 com- d)
pared with medium and long classes at feeding (Po 0.04) and Fat% 2.84 7 0.08a 3.177 0.08c 3.187 0.08c 0.003
Fat yield (kg/d) 1.147 0.08a 1.43 7 0.08c 1.40 7 0.08c 0.001
tended (P ¼0.10) to be lower at 1 h after feeding, this difference
Protein % 2.96 7 0.07 2.92 70.07 2.95 7 0.07 0.85
was not detectable after 2 h and was larger than initial GMPL. Protein yield 1.22 7 0.07 1.32 7 0.07 1.30 7 0.07 0.44
Changes in peNDF1.18 of refusal were similar between dairy farms (kg/d)
with different PS and increased with increasing GMPL of TMR and Fat-to-protein 0.94 7 0.02a 1.0770.02c 1.05 7 0.02c o 0.0001
ratio
with the progression of time.
Lactose % 4.69 7 0.04 4.79 70.04 4.73 70.04 0.06
Chewing activity was not affected by TMR PS and cows on Total solids % 11.93 7 0.14 12.29 70.14 12.317 0.14 0.09
average spent 340, 492, 825 min/d for eating, rumination, and SNF % 8.86 7 0.11 8.80 70.11 8.78 7 0.11 0.79
total chewing, respectively (Table 5). There was a tendency for
a,c
enhanced rumination on dairy farms that fed medium TMR PS Means within a row with different superscripts differ (Po 0.05).

relative to short and long (P ¼07).

Please cite this article as: Esmaeili, M., et al., Variation of TMR particle size and physical characteristics in commercial Iranian Holstein
dairies and effects on eating behaviour, chewing activity, and milk production. Livestock Science (2016), http://dx.doi.org/10.1016/j.
livsci.2016.07.003i
26 M. Esmaeili et al. / Livestock Science 191 (2016) 22–28

Table 7
Correlation coefficients (%) between physical characteristics, forage, and grain content of TMR (% of DM), and animal response variables on dairy farms with different TMR
particle size (n ¼ 15).

Variable GMPL1 peNDF82 peNDF1.182 S193 Forage Grain Milk fat Milk F:P4 Rumination Eating Chewing

GMPL 0.67** 0.83*** 0.22 0.10  0.51* 0.49T 0.53* 0.38 0.22 0.34
peNNF8 0.75*** 0.46T 0.61*  0.82*** 0.40 0.56* 0.48T 0.48T 0.60*
peNDF1.18 0.16 0.39  0.63* 0.38 0.64** 0.44 0.31 0.45T
S19 0.44T  0.29 0.51* 0.30 0.15 0.50* 0.46*
Forage %  0.75*** 0.26 0.41 0.22 0.78*** 0.71**
Grain %  0.43T  0.58*  0.33  0.62*  0.64**
Milk fat% 0.82*** 0.36 0.42 0.50T
Milk F:P 0.65** 0.30 0.54*

T
P r 0.10.
*
P r 0.05
**
P r0.01
***
P r 0.001
1
Geometric mean particle size length as calculated by ASAE (1995; method S424.1) with a sieving frequency of 1.1 Hz or approximately 1.1 shake per second.
2
peNDF8, peNDF1.18 ¼physically effective NDF calculated as dietary NDF content (% DM) multiplied by physical effectiveness factor determined as the DM proportion of
particles retained on 2 sieves (19 and 8 mm; pef8) or 3 sieves (19, 8, 1.18 mm; pef1.18), respectively, of the PSPS (Lammers et al., 1996; Kononoff, 2002).
3
Particles retained on the 19-mm sieve of PSPS ( 419 mm particles).
4
Ratio of milk fat to milk protein.

3.4. Milk production and composition
Milk production and 4% fat corrected milk (FCM) were not af-
fected by TMR PS on dairy farms, although cows fed medium and
coarse TMR produced numerically more milk and FCM (Table 6).
Milk fat yield and content were significantly affected by TMR PS;
cows fed with medium and coarse TMR had a greater fat percen-
tage and daily fat yield (P o0.01). Also, the ratio of milk fat-to-milk
protein (F:P) was greater in dairy herds fed with TMR with med-
ium and coarse than short PS (Po0.01). Lactose (P¼ 0.06) and
total solids (P¼ 0.09) tended to be greater on dairy farms fed with
coarse and medium rather than short TMR PS. Protein and SNF
were not affected by TMR PS in the current study.
3.5. Correlations among variables
Correlation coefficients between the important variables of this
study are shown in Table 7. The GMPL and peNDF1.18 correlated
with milk F:P, but not with chewing activity; peNDF8 correlated
with both milk F:P and chewing activity (P o0.01). The proportion
of particles 419 mm (r¼ 0.51) was the only variable that sig-
nificantly correlated with milk fat percentage (Po0.01) and ten-
ded to be correlated with eating (P ¼0.07) and total chewing time
(P ¼0.09). Other PSPS fractions and the percentage of alfalfa hay
and maize silage in the TMR (not reported in Table 7) were not
correlated with animal responses, except for silage percentage that
correlated with fat percentage (r ¼0.55 and P¼ 0.03). Interestingly,
grain percentage in the ration had a negative correlation (P o0.05)
with diet physical characteristics (ro  0.51) as well as eating
(r ¼ 0.62) and chewing activity (r ¼  0.64) and milk F:P
(r ¼ 0.58). Milk fat percentage did not correlate with chewing
activities, but milk F:P correlated (P o0.05) with rumination
(r ¼0.65) and total chewing time (r¼ 0.54).
4. Discussion
As expected, the physical form of TMR had a high variation
among dairy farms. The variation among dairy farms might be
attributed to differences in forage percentage, forage source, bo-
tanical characteristics of forages, as well as special practices con-
ducted by each dairy, such as using a silage face shaver. Sova et al.
(2014) suggested that increased surveillance of TMR composition,
not only of individual feed ingredients, may be helpful as a regular
component of feeding management to ensure delivery of TMR
with the intended nutrient composition to maintain production
and feed intake. Variation in TMR physical form may be proble-
matic for balancing rumen fermentation and buffering, which has
been shown to influence cow heath and profitability (Allen, 1997;
Enemark, 2009).
Rations in this study generally met recommendations for TMR
particle size distribution: 2–8% long particles, 30–50% medium and
short particles, and no more than 20% fine particles (Heinrichs and
Kononoff, 2002). The only discrepancy was in particles 4 19 mm,
which was slightly greater than recommended. The limitation for
particles 419 mm is to prevent sorting, which was usually re-
ported in previous studies for particles 419 mm (Leonardi et al.,
2005; DeVries et al., 2007, 2011). However, our previous in-
vestigation showed that this may not be the case for dairy farms in
Iran due to special conditions of the TMR and forages (Nasrollahi
et al., 2014). Indeed, coarse and palatable particles of immature
maize silage that were usually retained on the 19-mm screen in
Iranian dairy rations prevent sorting against 19-mm particles
(Kowsar et al., 2008; Akbari-Afjani., 2014; Nasrollahi et al., 2014).
The peNDF1.18 for all dairy farms was greater than the 21%
recommended by Mertens (1997). Dairy farms in the current study
used barley grain and cottonseed in the TMR, which contributed to
high levels of non-forage NDF in the diets. Due to this high amount
of non-forage NDF, the peNDF (expressed as pef  total NDF)
might have been higher than what is biologically effective in
motivating chewing and saliva, and preventing acidosis.
The most important observation of our study was improvement
in fat-related parameters of milk production with increasing TMR
PS along with numerically increasing milk yield. Increasing both
milk production and composition are the best outcome of good
management practices on dairy farms. This study shows that in-
creasing milk fat percentage along with preserving milk yield
could be achieved by increasing forage PS. Improvement in milk
fat with increasing TMR PS in the current study was in part in-
consistent with chewing activity data. In agreement with our data,
Kononoff et al. (2003) showed milk fat improvement without
improvement in chewing. They stated that factors such as the
acetate-to-propionate ratio and long-chain trans fatty acids are the
major components controlling milk fat production. Indeed, in-
creasing TMR PS along with homogenizing eating stabilized par-
ticle size in the rumen, allowing increased rumen fermentation
and acetate production and decreased trans fatty acid and pro-
pionate formation. These factors increase milk fat production
(NRC, 2001). Dairy farms with medium PS had the highest milk
production and milk fat percentage with a significant and

Please cite this article as: Esmaeili, M., et al., Variation of TMR particle size and physical characteristics in commercial Iranian Holstein
dairies and effects on eating behaviour, chewing activity, and milk production. Livestock Science (2016), http://dx.doi.org/10.1016/j.
livsci.2016.07.003i
 M. Esmaeili et al. / Livestock Science 191 (2016) 22–28
numerical difference from the short and coarse classes, respec-
tively. This effect may be related to the greater amount of material
retained on the 19-mm sieve in TMR of the medium class, because
the milk fat percentage in the current study was correlated with
this fraction, similar to a previous report (Kononoff, 2002).
The percentage of TMR retained on the 19-mm sieve of the
PSPS was a good indicator for maintaining cow health because of
the high and coincident relationship with milk fat and chewing.
The particle fraction 4 19 mm was the only variable that sig-
nificantly correlated with milk fat percentage. In an individual
feeding trial, Yang and Beauchemin (2007) observed a similar re-
lationship and reported that peNDF8 and the proportion of
particles 419 mm in TMR were more important variables than
peNDF1.18 regarding dairy cows’ response in chewing activity and
rumen fermentation. In agreement with this result, Nasrollahi
et al. (2012) showed grain source could interfere with peNDF1.18
due to retention on the 1.18-mm sieve of the PSPS without a
biological effect on improving chewing activity or rumen pH. Yang
and Beauchemin (2007) also reported the highest correlation be-
tween particles 419 mm and the proportion of acetate (r ¼0.53)
and propionate (r ¼  0.49), which may be the reason for the sig-
nificant relationship between particles 419 mm and milk fat
percentage in the present study. As result, parameters such as
peNDF8 and the proportion of particles 419 mm are the preferred
parameters for monitoring TMR physical form on commercial
dairy farms.
Grain percentage in the diet had a negative correlation with
dairy cow response regarding both milk F:P and chewing beha-
viour. This relationship was stronger than for diet physical char-
acteristics. Increasing the percentage of grain usually increased
diet fermentability as well as acid production in the rumen (Ei-
narson et al., 2004; Yang and Beauchemin, 2007, 2009), values
that are important in dairy cow behaviour, health, and milk fat
production (NRC, 2001; Stone, 2004).
5. Conclusions
Commercial dairy farms in this study had a high variation in
TMR PS, and cows on dairy farms which fed a TMR with coarse and
medium PS tended to have greater rumination activity and greater
milk fat percentage and yield. The peNDF8 and proportion of
particles 4 19 mm were better indicators for milk fat percentage
and chewing activity than peNDF1.18. Also, grain percentage in the
diet had a negative, but strong correlation with milk F:P and
chewing behaviour. There is a critical need to add the role of for-
age PS and grain level in modeling dairy cow nutrition against
milk fat depression in commercial dairy farms.
Acknowledgement
Authors thank Ali Sadeghi-Sefidmazgi (Isfahan University of
Technology, Iran) for his statistical assistance and D.J.R. Cherney
(Cornell University), C.M. Jones (Penn State University), E.K. Ins-
keep (West Virginia University), and M. Hutjens (University of Il-
linois) for their valuable editions. Also, we thank the workers,
engineers, and managers of Iranian dairy farms for helping us in
data collection; they are working in hard conditions with a small
amount of payment and producing milk for us.
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Please cite this article as: Esmaeili, M., et al., Variation of TMR particle size and physical characteristics in commercial Iranian Holstein
dairies and effects on eating behaviour, chewing activity, and milk production. Livestock Science (2016), http://dx.doi.org/10.1016/j.
livsci.2016.07.003i