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Characteriztion of Quercus persica, Quercus infectoria and

Quercus libani as ruminant feeds

Article  in  Animal Feed Science and Technology · January 2008

DOI: 10.1016/j.anifeedsci.2007.02.009

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Animal Feed Science and Technology

140 (2008) 78–89

Characteriztion of Quercus persica,

Quercus infectoria and Quercus libani
as ruminant feeds
M. Yousef Elahi, Y. Rouzbehan ∗
Animal Science Department, Faculty of Agriculture, Tarbiat Modares University,
P.O. Box 14115-336, Tehran, Iran

Received 17 August 2006; received in revised form 7 February 2007; accepted 21 February 2007

Abstract

The nutritive value of three species of oak trees leaves was assessed by chemical analysis as well
as by an in situ and in vitro gas production method. The chemical composition (g/kg DM basis) of
three species, Quercus persica, Q. infectoria and Q. libani were 951, 927, 946 organic matter (OM);
115, 92, 123 crude protein (CP); 532, 540, 512 neutral detergent fiber (NDFom); 317, 300, 331 acid
detergent fiber (ADFom); 98, 103, 95 lignin (lignin(sa)); 8.9, 11.7, 19.2 water soluble carbohydrates
(WSC); 78, 115, 104 total phenols; 73, 109, 100 total tannins (TT); 14, 15, 12 condensed tannin (CT);
and 46, 87, 62 hydrolysable tannin (HT). Protein precipitable phenolics (PPP) were respectively
160, 190 and 230 (g/kg total phenols). Effects of tannins on in vitro gas production, in vitro organic
matter digestibility (OMD), metabolisable energy (ME) and effective DM digestibility (ED) were
assessed by polyethylene glycol (PEG) tannin bioassay. Among species, Q. Libani had the highest
gas production. This species also had the highest (P<0.05) increase in gas production, OMD and
ME due to the addition of PEG. Using this in situ method the soluble component (A), insoluble but
fermentable component (B), the degradation rate of B (c), the potential degradability (A + B) and the
effective degradability (ED) of the oak leaves were influenced (P<0.05) by species, PEG treatment

Abbreviations: ADFom, ash-free acid detergent fiber; CP, crude protein; CT, condensed tannin; DM, dry matter;
ED, effective degradability of DM; HT, hydrolysable tannins; lignin(sa), lignin determined by solubilisation of
cellulose with sulphuric; ME, metabolisable energy; MW, molecular weight; NDFom, ash-free neutral detergent
fiber; NTP, non-tannin phenol; OM, organic matter; OMD, OM digestibility; PEG, polyethylene glycol; PPP,
protein precipitable phenolics; TPH, total phenols; TT, total tannin; WSC, water soluble carbohydrates
∗ Corresponding author. Tel.: +98 21 44194911; fax: +98 21 44196524.

E-mail address: rozbeh y@modres.ac.ir (Y. Rouzbehan).

0377-8401/$ – see front matter © 2007 Elsevier B.V. All rights reserved.
doi:10.1016/j.anifeedsci.2007.02.009
 M. Yousef Elahi, Y. Rouzbehan / Animal Feed Science and Technology 140 (2008) 78–89 79

and interaction of species and PEG treatment. Adding PEG to Quercus species suggests that these
feeds have potential as small ruminant feeds.
© 2007 Elsevier B.V. All rights reserved.

Keywords: Oak leaves; Nutritive value; In situ degradability; In vitro gas production; Goats

1. Introduction

Oak leaves and twigs are often grazed by ruminants or harvested for use as livestock
feed during feed shortages (Singh et al., 1996). Approximately 3 million ha of forest are
covered by various oak species, mainly dominated by Quercus persica, Quercus infectoria
and Quercus libani, in the north-west of Iran (Fatahi, 1995). In this region, oak leaves are an
important source of forage for goats during periods of the year when quality and quantity
of pasture herbages is limited. However, Quercus species have been reported to contain
high levels of tannins in both hydrolysable (Makkar, 2003) and condensed (Makkar et al.,
1991) forms. Therefore, the value of these leaves as feeds for ruminants is offset by their
potentially negative effects on protein utilization, and the risk of toxicity when intake is high
(Garg et al., 1992). Polyethylene glycol (PEG), a non-nutritive synthetic polymer, has a high
affinity to tannins and can make them inert by forming tannin–PEG complexes (Makkar et
al., 1995). PEG can also liberate protein from tannin–protein complexes (Barry and Manley,
1986) and/or prevent their formation. Therefore, PEG has been used to mitigate adverse
effects of tannins on rumen fermentation.
In situ rumen disappearance and in vitro gas production techniques are useful for rapid
screening of feeds to assess their potential as feed energy sources for ruminants (Preston,
1995). However, the gas production technique has generally proved to be more effective
than rumen in situ techniques to determine the nutritive value of feeds containing inhibitory
compounds, such as tannins. For example, Khazaal et al. (1994) showed that while physical
binding of tannins to substrate could be detected in a rumen in situ incubation, effects such
as toxicity to microbes and binding to enzymes are diluted and difficult to detect. In contrast,
the in vitro gas production technique, being a closed system with a limited supply of rumen
fluid, may more reliably detect effects of inhibitory compounds in feeds on their potential
ruminal digestion.
No information is available on Q. persica, Q. infectoria and Q. libani as ruminant feeds,
and so this study evaluated the chemical composition, in situ degradation and in vitro gas
production.

2. Materials and methods

2.1. Quercus species

Samples consisted of leaves of three indigenous Quercus species, being Q. persica, Q.

infectoria and Q. libani. Samples were harvested by hand during the summer at several loca-
tions in the NW of Iran. Branches were randomly sampled from at least 10 plants per species.
80 M. Yousef Elahi, Y. Rouzbehan / Animal Feed Science and Technology 140 (2008) 78–89

Leaves were removed from branches, pooled to five samples per species and air dried in the
shade to minimize changes in tannin content and activity (Makkar and Singh, 1991b).
Water soluble carbohydrate (WSC) was measured using the anthrone method (MAFF,
1982).

2.2. Chemical analysis

Standard methods as described in AOAC (1990) were used for determination of dry
matter (DM, method 930.15), ash (method 924.05) and N (method 984.13). Ash-free neutral
detergent fibre (NDFom) was determined using sodium sulfite according to the method of
Van Soest et al. (1991) and ash-free acid detergent fibre (ADFom, method 973.18) was
determined based on AOAC (1990). Lignin(sa) was determined by solubilisation of cellulose
with sulphuric acid as described by Robertson and Van Soest (1981).
Samples were analysed for total phenols (TPH), total extractable tannin (TT), con-
densed tannins (CT), hydrolysable tannins (HT) and protein precipitable phenolics (PPP),
as described below.

2.2.1. Total phenols

Dried plant material (200 mg) was extracted with acetone:water (10 ml; 70:30, v/v)
in ultrasonic bath for 20 min. Contents were centrifuged (4 ◦ C, 10 min, 3000×g) and
the supernatant was kept on ice until analysis. Total phenols were determined with the
Folin–Ciocalteau reagent and detected at 725 nm (Makkar, 2000). A calibration curve
was prepared using tannic acid (Merck GmbH, Darmstadt, Germany). Total phenols were
calculated as tannic acid equivalents and expressed as equiv. g/kg DM.

2.2.2. Tannins
Condensed tannins were measured by the HCL-butanol method (Makkar, 2000). An
aliquot from the above acetone:water extract (0.5 ml; although this extract occasionally
needed diluting with the extractant, acetone:water, if final absorbance at 550 nm exceeded
0.6 absorbance units) plus HCL-butanol (3 ml) and ferric ammonium sulphate (0.1 ml)
reagents were heated in a boiling water bath for 60 min. Absorbance was read at 550 nm.
The colorimetric data (in absorbance units) were converted to leucocyanidin equivalents
based on the assumption that the colour yield of condensed tannins, E1%,550 nm , is 460
(Porter et al., 1986).
Non-tannin phenols (NTP) were determined using absorption to insoluble
polyvinylpyrrolidone. The insoluble polyvinylpyrrolidone (PVPP; 100 mg) was weighted
into 100 mm×12 mm test tubes. Distilled water, 1 ml, and then 1 ml tannin containing
extract were added and vortexed. The tube was kept at 4 ◦ C for 15 min, vortexed again,
then centrifuged (3000×g) for 10 min and supernatant collected. The phenolic content of
the supernatant was measured by the Folin-Ciocalteau reaction and this was accepted as the
NTP (Makkar, 2000).
Total tannins were calculated as the difference between TPH and NTP. Hydrolysable
tannins were analysed using Rhodanine assay according to Makkar (2000). The results
were expressed as gallotannin. Protein percipitable phenilics were determined according to
Makkar (2000), and results were expressed as tannic acid equivalent.
 M. Yousef Elahi, Y. Rouzbehan / Animal Feed Science and Technology 140 (2008) 78–89 81

2.3. Gas production

Rumen fluid for the in vitro digestibility tannin bioassay was obtained from three
healthy mature Markhoz male goats with live weight of 46 ± 1.5 kg fitted with perma-
nent 70 mm rumen cannulae that were fed a daily ration of a mixture of 800 g lucerne
hay and oak leaves (50:50) and 375 g concentrates (500 g/kg barely grain, 250 g/kg
wheat bran, 210 g/kg soybean meal and 40 g/kg minerals and vitamins premix which
contained (per kg) 185 g Ca, 104 g Mg, 2.25 g Co, 44.0 g Mn, 36.4 g Zn, 1.3 g I,
10,000,000 iu retinol, 2,000,000 iu vit. D3 and 40,000 iu ␤-tocopherol) divided into
equal meals at 8:00 and 16:00 h daily. Goats had free access to water throughout the
experiment.
Rumen fluid was obtained from the three goats in the morning before feeding (07:00),
flushed with CO2 , filtered through three layers of cheesecloth and mixed (1:2, v/v)
with an anaerobic mineral buffer solution as described by Makkar et al. (1995) and
revised by Makkar (2000). Preparation of an in vitro mineral buffer media for the gas
test was completed out as described by Menke and Steingass (1988). Reduced buffer
medium composition, per litre, was NaHCO3 , 70.0 g; NH4 HCO3 , 4.00 g; Na2 HPO4 , 5.7 g;
KH2 PO4 , 6.2 g; MgSO4 ·7H2 O, 0.6 g; Na2 S, 0.52 g; CaCl2 ·H2 O, 13.2 g; MnCl2 ·4H2 O,
10.00 g; CoCl2 ·6H2 O, 1.00 g and sodium resazurin, 0.01 g and, 60 ml freshly prepared
reduction solution containing 580 mg Na2 S·9H2 O and 3.7 ml 1 M NaOH. The mixture
was kept stirred, under CO2 flushing at 39 ◦ C, using a magnetic stirrer fitted on a hot
plate.
Effects of tannins on in vitro OM digestibility (OMD) were assessed by incubat-
ing approximately 375 mg (DM) of triplicate test feed samples with or without 750 mg
polyethylene glycol (PEG), molecular weight (MW, 6000, Merck Schuchardt, Hohenbrunn,
Germany). Feed samples were incubated in 100 ml glass syringes based on Menke and
Steingass (1988) procedures. The PEG tannin bioassay was according to Makkar et al.
(1995) as revised by Makkar (2000). Petroleum jelly was applied to the piston to ease
movement and prevent escape of gas. Syringes were pre-warmed (39 ◦ C) for 1 h before
addition of 30 ± 0.5 ml of rumen buffer mixture into each syringe, and incubated in a water
bath maintained at 39 ± 0.1 ◦ C as described by Blümmel and Orskov (1993). Syringes
were gently shaken every hour during the first 8 h of incubation. Gas production read-
ings (ml) were recorded at 2, 4, 6, 8, 10, 12, 16, 24, 48, 72 and 96 h for PEG treated and
untreated samples. Total gas values were corrected for blank and a hay standard with a
known gas production. Cumulative gas production data were fitted to the exponential equa-
tion Y = b (1 − e−ct ), where b is the gas production from the fermentable fraction (ml), c
the gas production rate constant for b, t the incubation time (h) and Y is the gas produced at
time t.
Feed OMD (g/kg OM) and metabolisable energy (ME) in MJ/kg DM were estimated
by equations of Menke and Steingass (1988), based on 24 h gas production (Gas, ml)
and CP content (g/kg DM) as: OMD (g/kg OM) = 148.8 + 8.89 GAS + 4.5 CP + 0. 651 XA
and ME (MJ/kg DM) = 2.20 + 0.136 GAS + 0.057 CP + 0.0029 CP2 . Where OMD is OM
digestibility, ME is metabolisable energy; CP is crude protein in g/100 g DM; XA ash in
g/100 g DM; and GAS the net gas production (ml). Net gas production data were converted
from 375 to 200 mg after 24 h of incubation.
82 M. Yousef Elahi, Y. Rouzbehan / Animal Feed Science and Technology 140 (2008) 78–89

2.4. In situ degradability

Six rumen-fistulated Markhoz goats (live weight 46 ± 1.5 kg) were used to determine the
rate of degradability of DM (AFRC, 1992) from oak leaves. Three goats which were fed the
PEG-treated diet were used to evaluate the PEG-treated oak leaves and three, which were
fed the non-treated diet, were used to assess the non-treated diet. PEG treatment consisted
of spraying lucerne hay and dried leaves with a solution of 90 g of PEG 6000 in 300 ml of
water per kg of DM (Ben Salem et al., 1999), which occurred when preparing the diets and
the in situ bag method. Goats were fed 1.18 kg/day of a ration consisting of lucerne hay, oak
leaves (mix of three oak spieces), wheat bran and barley grain (50:50) with a ratio of forage
to concentrate of 60:40 (DM basis), which was calculated to ME at their maintenance level.
Goats were adapted to the diet for 10 days.
In situ bags were made from a Dacron material (21 cm×10 cm) with a pore size of 45 ␮m
(AFRC, 1992). All samples of feeds were dried and milled through a 4.0 mm sieve. Then,
5 g of each sample was put in the in situ bags and incubated at the same time in the rumen
for 3, 6, 12, 24, 48, 72 and 96 h. In each goat, one bag was used for each time interval. Bags
were attached on semi-rigid stalks to ensure immediate insertion within the liquid of the
rumen contents while allowing free movement. After withdrawing the bags from the rumen,
they were washed in a washing machine for 1 h using cold water and dried for 48 h at 50 ◦ C.
The degradability value at t = 0 was obtained by washing two bags in a washing machine
for 1 h using cold water. For each bag, the residue was analyzed for DM. Degradability at
each incubation time was calculated by taking the values obtained from the three bags (i.e.,
n = 3). The ruminal degradability (Y) of DM at time (t) was obtained from an exponential
curve of the type:
Y = a + b(1 − e(−ct) )
This was fitted to the experimental data by iterative regression analysis (Ørskov and
McDonald, 1979). In this equation, the constant a represents the soluble and very rapidly
degradable component and b represents the insoluble but potentially degradable component
which degrades at a constant fractional rate (c) per unit time. The effective degradability
of DM in each species was then estimated (Ørskov and McDonald, 1979) by the equation:
effective degradability (g/kg DM) = a + bc/c + k. In this equation, k refers to the fractional
outflow rate of small particles from the rumen. A value of 0.05 fraction/h was used for k.

2.5. Statistical analysis

Data on chemical and tannin composition were subjected to analysis using the Gen-
eral Linear Model (GLM) procedure of SAS (2001), based on the statistical model:
Yij = μ + Si + eij , where Yij is the general observation on chemical composition and tan-
nin composition, μij the general mean, Si the ith effect of oak species on the observed
parameters and eij the standard error term. Means were tested using Duncan test. For in
situ, in vitro gas production and digestibility estimates data, the following statistical model
was fitted:
Yijk = μ + Si + Pj + (SP)ij + Eijk
 M. Yousef Elahi, Y. Rouzbehan / Animal Feed Science and Technology 140 (2008) 78–89 83

where Yijk is the observation on in situ, gas production and digestibility estimates, Si the ith
effect of oak species and Pj the jth effect of PEG on the observed parameters. The (SP)ij
term represents ith and jth interaction effects of species and PEG on in situ and gas in
vitro production, and Eijk the standard error term common for all observations. When the
interaction between species and PEG, either in gas production or in situ degradability, was
significant (i.e., P<0.05) the differences between control and PEG within oak species were
examined using a T-test.

3. Results

3.1. Chemical composition and phenolics

Quercus infectoria leaves had the lowest OM, CP and ADF (Table 1), but contained the
highest NDFom and lignin(sa) versus other species. In all Quercus spp., the level of HT was
high. Among the oak leaves, Q. infectoria had the highest TPH, TT, CT and HT content
(P<0.05). However, the biological value of tannins as PPP (g/100 g of total phenolics) was
higher (P<0.05) in Quercus libani versus other species.

3.2. Gas production

Gas production characteristics (i.e., b, c, ME and OMD) during the fermentation period
are in Table 2. The volume produced from the insoluble but fermentable (i.e., b) part of
the feed ranged from 35.9 to 36.8 ml, respectively. These gas volumes were produced at
a rate varying from 0.059 to 0.067 h−1 for the untreated samples and a rate varying from
0.054 to 0.060 h−1 for treated samples. Although Q. libani leaves have higher (P<0.05)

Table 1
Chemical composition and phenolics (g/kg DM or as stated) of oak species
Q. persica Q. infectoria Q. libani S.E.M.
Chemical composition
DM 939 b 943 a 946 a 0.4
OM 951 a 927 c 946 b 0.9
CP 115 b 92 c 123 a 0.1
ADFom 317 ab 300 b 331 a 0.7
NDFom 532 a 540 a 512 b 0.4
Lignin(sa) 98 b 103 a 95 b 1.3
WSC 8.9 b 11.7 b 19.2 a 0.51
Phenolics
TPH 78 c 115 a 104 b 0.2
TT 73 c 109 a 100 b 0.2
CT 14 b 15 a 12 c 0.05
HT 46 c 87 a 62 b 0.1
PPP 16 c 19 b 23 a 0.4
Means in the same row with different letters differ (P<0.05).
 84
M. Yousef Elahi, Y. Rouzbehan / Animal Feed Science and Technology 140 (2008) 78–89
Table 2
Gas production characteristics and estimated parameters and in situ dry matter disappearance in oak species treated (PEG) and non-treated with PEG (control)
Q. persica Q. infectoria Q. libani Significance

Control +PEG Control +PEG Control +PEG S.E. Species PEG Species×PEG
Gas production
b 36.6 b 40.36 a 35.94 a 39.12 b 36.78 b 43.13 a 0.341 * * *
c 0.059 a 0.054 a 0.059 a 0.06 a 0.067 a 0.057 a 0.0011 * * *
ME 6.82 7.19 6.66 6.86 7.36 7.71 0.071 * * NS
OMD 441 468 430 450 466 501 3.7 * * NS
In situ disappearance of DM
A 153 b 161 a 160 b 179 a 201 b 203 a 0.3 * * *
B 207 b 232 a 252 b 301 a 281 b 319 a 0.3 * * *
A+B 360 b 393 a 418 b 480 a 485 b 518 a 0.3 * * *
C 0.024 b 0.041 a 0.03 a 0.032 a 0.023 a 0.028 a 0.0021 * * *
ED 220 b 254 a 264 b 292 a 293 b 313 a 0.2 * * *
b: insoluble but fermentable fraction (ml); c: rate constant of gas production during incubation (ml/h); OMD: organic matter digestibility (g/kg DM); ME: metabolisable
energy (MJ/kg DM); A: water-soluble fraction (g/kg DM); B: insoluble but fermentable fraction (g/kg DM); C: the degradation rate of B (/h); A + B: the potential
degradability (g/kg DM); ED: the effective degradability of dry matter calculated for an outflow rate of 0.05/h (g/kg DM). Means with different letters within species
differ (P<0.05); NS: non-significant.
 M. Yousef Elahi, Y. Rouzbehan / Animal Feed Science and Technology 140 (2008) 78–89 85

gas production characteristics (i.e., b, c, ME and OMD) than other species, the difference
is small. Addition of PEG increased (P<0.05) gas production characteristics b, OMD and
ME content in all Quercus species. However, responses in b and c due to addition of
PEG in vitro were influenced significantly (P<0.05) by interaction of species and PEG
treatment.

3.3. In situ DM disappearance and estimated parameters

Characteristics of the DM disappearance of the oak leaves are in Table 2. The soluble
component (A), insoluble but fermentable component (B), the degradation rate of B (c),
the potential degradability (A + B) and the effective degradability (ED) of the oak leaves
were influenced (P<0.05) by species, PEG treatment and interaction of species and PEG
treatment. However, variable responses in A, B, (A + B), c and ED due to addition of PEG
occurred due to interactions of species and PEG treatment. In the untreated and treated
samples, the DM disappearance characteristics of Q. libani were higher (P<0.05) than
those in other species.

4. Discussion

4.1. Chemical composition and phenolics

The variation in chemical composition among these species of Quercus may be partly
due to the genotypic factors that control accumulation of forage nutrients (Minson, 1990). In
our oak leaves, the CP content was more than 80 g/kg DM (range: 92–123 g/kg DM) which,
according to Norton (1998), should provide ruminal ammonia levels above the minimum
required by rumen microorganism to support optimum growth. Makkar and Singh (1991b)
reported a similar range of CP content in mature Q. incana, Q. semecarpifolia, Q. serraata,
Q. ilex and Q. glauca. However, Kamalak et al. (2004) obtained lower CP content in Q.
branti, Q. coccifera, Q. ceris, Q. libari and Q. infectoria than our Qercus spp. The ADFom
and lignin(sa) content of our oak species were lower than in Q. incana, Q. semecarpifolia,
Q. serraata, Q. ilex and Q. glauca (Makkar and Singh, 1991b), but higher than in Q. libari
(Kamalak et al., 2004). The level of NDFom in our experiment was lower than in Q. incana,
Q. semecarpifolia and Q. serraata than Q. libari (Kamalak et al., 2004). There was a
relationship between TPH and TT which is similar to findings of Makkar et al. (1993) who
noted a high positive correlation between content of total tannins and total phenolics. Levels
of TPH and total tannins in the experimental species were higher than in Q. incana (Singh et
al., 2005), Q. hartwissiana (Yildiz et al., 2005), Q. coccifera (Ben Salem et al., 2003, 2005),
similar to Q. rotundifolia (Khazaal et al., 1994), but lower than Q. coccifera (Khazaal et
al., 1994). Condensed tannins in our oak leaves were similar to Q. hartwissiana (Yildiz et
al., 2005), Q. coccifera (Khazaal et al., 1994) and Q. suber (Gasmi-Boubaker et al., 2005),
but lower than Q. coccifera (Ben Salem et al., 2003, 2005); Q. incana (Singh et al., 2005),
Q. cercis (Canbolat et al., 2005). The level of HT is high in Quercus spp. Similarly, some
groups have reported that oak leaves are rich in HT (Garg et al., 1992; Makkar, 2003).
However, others noted low levels of HT in oak leaves (Yildiz et al., 2005; Singh et al.,
86 M. Yousef Elahi, Y. Rouzbehan / Animal Feed Science and Technology 140 (2008) 78–89

2005). The protein precipitation assay (PPP) measures tannin activity as it depends on the
functional properties of tannins. Among oak leaves, the highest tannin activity was in the
Q. libani. The variations between our oak leaves and other oak species in the chemical
composition and phenolics contents is probably due to any or all of the vegetative stage
(Makkar and Singh, 1993), method of storage (Makkar and Singh, 1993), drying conditions
(Makkar and Singh, 1991a), species (Makkar and Singh, 1991b; Makkar et al., 1991) and
habitat (Goncalves-Alvim et al., 2004).

4.2. Gas production

The OMD in Q. libani were higher than those in other species, possibly because Q. libani
contains higher WSC and CP levels which are vital substrates for ruminal microorganisms
growth (Van Soest, 1994). Also, the lower NDFom and lignin(sa) content in Q. libani versus
other species may result in increased OMD (Van Soest, 1994; Makkar, 2003).
Increased gas production and OMD due to addition of PEG in vitro suggests a negative
influence of tannins on digestibility (Makkar, 2003). Inactivation of tannins through PEG
binding increases availability of nutrients resulting in increased microbial activity and gas
production (Makkar, 2003). Similar results were obtained by Khazaal et al. (1993) when
using PEG as the phenolic binding agent in conjunction with an in vitro gas technique for
assessment of secondary compounds in browse.
PEG treatment caused different increments in gas production among Quercus spp.,
with Q. libani the highest. These variable responses of gas production among our Quer-
cus spp. could be due to variations in tannin activity (PPP, Table 1) between Quercus
spp. Values obtained using a protein precipitation assay better relate to the nutritional
values of tannin-rich feeds (Hagerman and Butler, 1989; Makkar, 1989). Tannin activ-
ity has been reported to vary among forage species due to the in functionality of tannin
and chemical structure (Dalzell and Kerven, 1998), degree of polymerization (Schofield
et al., 2001), biochemical processes (Wong, 1973) and to the tannin structure–biological
activity relationship (Haslam, 1998). Similarly, Rubanza et al. (2005) reported variable
responses in gas production characteristics, due to addition of PEG, between Acacia
spp.

4.3. In situ degradability

On the basis of the DM disappearance characteristics with and without PEG, the species
were ranked as Q. libani > Q. infectoria > Q. persica. However, on the basis of the concen-
trations of parameters such as WSC, CP, NDFom, lignin(sa) and CT, the ranking would be
as Q. libani > Q. persica > Q. infectoria.
Effects of PEG had variable responses on the proportion centage of increased ED of
DM disappearance (115.5, 110.6 and 106.8 for Q. persica, Q. infectoria and Q. libani,
respectively). These results did not parallel with tannin activity (PPP, Table 1), suggesting
that this in situ DM disappearance method did not measure the biological effects of tannins
in these Quercus spp. A similar conclusion was reached by Vitti et al. (2005), who observed
that in situ results did not correlate with any of the phenolics compound data, and the
increased gas production, due to PEG inclusion in three Brazilian fodder legumes.
 M. Yousef Elahi, Y. Rouzbehan / Animal Feed Science and Technology 140 (2008) 78–89 87

5. Conclusion

On the basis of WSC, CP, NDFom, lignin(sa), CT, in vitro gas production and DM
degradation parameters, oak leaves of Q. libani species were judged to be nutritionally
best. Addition of PEG, which has a relatively low cost in Iran, inactivated effects of tannins
and increased gas production. It appears that a PEG/gas production assay is a useful first
screen for tannin-rich feeds. However, further research is needed to assess their impacts on
animal performance.

Acknowledgment

The authors are grateful to Prof. H. Makkar for his helpful suggestions and for correcting
the manuscript.

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