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International Journal of Environment, Agriculture and Biotechnology (IJEAB) Vol-2, Issue-6, Nov-Dec- 2017

http://dx.doi.org/10.22161/ijeab/2.6.9 ISSN: 2456-1878

Productive and Reproductive Traits of Sheep


Fed Acacia saligna Leaves-Based Diets
Mohamed N. El-Gendy2, Mohamed M. Anwar2, Wael G. Fahmy1, Samir Z. El-
Zarkouny1,3, Nesrin M. Hashem1, Adel N. M. Nour El-Din1, Marwa F.A. Attia1, El-Saeed
A. El-wakeel2, Moustafa M. Zeitoun1,*
1
Department of Animal and Fish and Production, Faculty of Agriculture, University of Alexandria, Egypt.
2
Animal Production Research Institute, Agricultural Research Center, Dokki, Giza, Egypt.
3
Department of Arid Land Agriculture, Faculty of Meteorology, Environment and Arid Land Agriculture, KAU, KSA

Abstract Investigating effects of partial (50%) or total and crop residues. In Egypt, the primary constraints to
(100%) substitution of clover hay by tannins-rich plant livestock production are scarcity and fluctuating quantity
(Acacia saligna) on productive and reproductive and quality of the year-round feed supply. The gap
performance of ewe lambs was the main goal of this between available and required amounts of animal
study. Two experiments were conducted: first focusing on feedstuffs in Egypt was estimated to be 3.5 million tons.
digestibility and N balance using 9 Barky rams (live body There are two alternative strategies to overcome the
weight, 43 2.5 kg) where animals were randomly problem of feeds shortage in Egypt. The first is to
divided into 3 groups (n = 3); control (C), 50% Acacia maximize utilization of agricultural and industrial
(AS50%) and 100% Acacia (AS100%); second focusing on byproducts and there are several studies focused on this
productive and reproductive performance of ewe-lambs point. The second strategy is seeking alternative
(n=18) where animals were divided into three groups nonconventional feed resources e.g. tannins-rich plants or
(n=6); C, AS50% and AS100%. This experiment started 2 shrub legumes, which have high protein contents and are
months before mating and continued till weaning. Dry potentially promising to overcome nutrient deficiencies.
matter intake decreased (P<0.05) linearly due with The multipurpose trees and tannins-rich plants represent
treatment. The digestion coefficients of dry matter (DM), an important fodder reserve for ruminants in periods of
organic matter (OM), crude protein (CP), neutral feed scarcity and play vital role in bridging the wide gap
detergent fiber (NDF) and acid detergent fiber (ADF) between supply and demand of feeds [1]. Use of these
were lower (P<0.05) in treated than C. The nitrogen forages in animal diets may participate in reducing the
intake and urinary nitrogen were reduced (P<0.05) by shortage of animal feed resources, enhance the fertility
treatment, while fecal nitrogen increased (P<0.0%) with performance of animals and subsequently increase milk
treatment. No change was found in conception rates and meat production in tropic regions. The shrub foliages
among the three groups whereas fertility rates and already play an important role in ruminant feeding
lambing rates were higher in the treated compared to the systems in Mediterranean and many tropical
control. AS100% reduced (P<0.05) total protein and blood environments around the world [2- 5].
urea nitrogen (BUN) compared to other groups. No To efficiently use alternative feed resources like tannins-
differences in progesterone concentration were found rich plants, their effects on various aspects need to be
among groups. AS50% resulted in higher (P<0.05) milk considered. This needs more research to be done for fair
yield than other groups. Treatment decreased (P<0.05) judgment within different points of view. Therefore, the
milk fat percentage, whereas didnt change protein and objectives of this study were to assess effects of the
lactose. Therefore, partial replacement of acacia leaves partial or total replacement of clover hay by acacia leaves
in sheep diets could be beneficial for productive and as an alternative feed resource on the dry matter intake,
reproductive performance. nutrients utilization, ruminal fermentation profiles,
Keywords Feed additives, digestibility, rumen reproductive and productive performance of Barky sheep.
fermentation, blood metabolites, prolificacy.
II. MATERIAL AND METHODS
I. INTRODUCTION This study was carried out at the Nubaria Research
The major limitation to ruminants production in many Station, Animal Production Research Institute,
tropical regions is poor nutrition, which is characterized Agricultural Research Center, Ministry of Agriculture and
by low nitrogen and high fiber content in native grasses Land Reclamation, Dokki, (Exp.1) and the Research Farm

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International Journal of Environment, Agriculture and Biotechnology (IJEAB) Vol-2, Issue-6, Nov-Dec- 2017
http://dx.doi.org/10.22161/ijeab/2.6.9 ISSN: 2456-1878
of the Department of Animal and Fish Production, Acacia leaves were collected weekly during both
Faculty of Agriculture, University of Alexandria (Exp. 2), experimental periods in order to offer as fresh materials
Egypt. Two experiments were designed to investigate the for animals. Extra samples of acacia leaves were collected
potential replacement of clover (Trifolium alexandrina) monthly during a year to analyze total phenols (TP), total
hay by acacia (Acacia saligna, AS) leaves either partially tannins (TT) and condensed tannin (CT). The collection
(50%, AS50%) or totally (100%, AS100%). First experiment region was located at latitude
was designed to investigate digestibilty and nitrogen of 30.91and longitude: 29.68. The region climate was
balance trial. However, the second experiment focused on semi-arid Mediterranean climate, which is characterized
the productive and reproductive performance of ewe- by a brief mild and rainy winter and long warm summer
lambs. months with no rain. The chemical composition and
tannins content of the commercial concentrate mixture,
Acacia samples collection clover hay and acacia leaves for both experiments are
presented in Table 1.

Table.1: Chemical composition and tannins content of the concentrate mixture, clover hay and Acacia leaves
(Based on dry matter)
Item (%) Concentrate Clover hay Acacia leaves
OM 88.5 89.3 89.6
CP 14.1 12.5 13.2
EE 5.8 1.0 4.8
NDF 38.3 64.2 64.9
ADF 10.6 55.8 60.6
Tannins content
TP* - 10.2 42.1
TT* - 1.6 27.1
CT** - 2.0 72.0
OM: Organic matter, CP: Crude protein, EE: Ether extract, NDF: Neutral detergent fiber, ADF: Acid detergent fiber. *TP:
total phenols (eq-g tannic acid kg-1DM), *TT: total tannins (eq-g tannic acid kg-1DM), **CT: condensed tannins (eq-g
leucocyanidin kg-1DM).

Experiment 1 feces and urine were collected for the purpose of


Animals and experimental design measuring nutrients digestibility and N balance.
Nine mature Barky rams (mean live body weight 432.5 To measure apparent total tract nutrients digestibility, two
kg) were randomly divided into three equal groups (n = 3) representative samples of feces 10 % of the total quantity)
as control (C), AS50% and AS100% according to live body were collected daily from each animal; one of the two
weight. Animals were housed in well-ventilated shade and samples was sprayed with citric acid (10%) and stored
adapted to the treatment feeds for 4 weeks before being under -20C, the second was used for determination of the
subjected to the digestibility trial and had free access to feces DM. Immediately, at the end of collection period
fresh water. The animals were fed in groups on a stored feces samples during 5 days for each animal were
commercial concentrate feed mixture and roughages pooled, mixed well and a sample was obtained for further
according to NRC [6]. The acacia leaves were offered analyses. Representative portions of feces were dried in a
fresh daily as partial or total replacement to the clover hay forced air-oven at 50C for 48 h and then ground to pass
by 1.0 kg and 2.0 kg/h/d, respectively, while the clover through a 1 mm-screen and stored at -20C thereafter
hay was offered to the control and partial replacement until analysis.
group by 1.0 and 0.5kg/h/d, respectively. The commercial The urine was collected daily throughout the collection
concentrate feed mixture was offered to all groups by 0.75 period in plastic buckets containing 100 mL of H2SO4
kg/h/d for all groups. After familiarization to the diets, the (10%). Also, representative samples (10% of total
animals were kept in metabolic cages for 8 days of which volume) were collected daily from each animal and kept
3 days for adaptation and 5 days for samples collection. in dark bottles. At the end of collection period, urine
Experimental animals were offered concentrate and samples from each animal were mixed well and obtained
roughages individually in buckets twice daily at 8:00 and a sample, which kept under -20C until analysis. Upon
14:00 hr. During the collection period, animals were analysis, thawed urine samples were centrifuged at 2000
individually kept in metabolic cages and feed refusals, rpm for 20 min and sub-samples were analyzed for

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International Journal of Environment, Agriculture and Biotechnology (IJEAB) Vol-2, Issue-6, Nov-Dec- 2017
http://dx.doi.org/10.22161/ijeab/2.6.9 ISSN: 2456-1878
Kjeldahl N [7]. Ruminal fluid samples were obtained PGF2 received a second injection of PGF2 (0.5 ml of
from each animal via a stomach tube before morning estroPLAN with the concentration of 125 g
feeding for two consecutive days. The ruminal fluid Cloprostenol) after 12 days from the first injection then
samples were separated from the feed particles through submitted to estrus detection twice daily using mature
four layers of cloth sheets and then stored under -20oC for teaser ram at 7.00 and 19.00 hr for 3 days. Ewe-lambs
VFA and ammonia N analyses. which were mounted by teaser rams were mated by fertile
rams. A blood sample was taken from each ewe-lamb at
Experiment 2 the days of injection, estrus, 5, 10 and 20 after estrus for
Animals and experimental design determination of BUN and progesterone.
Eighteen ewe-lambs (live body weight of 352.5 kg and Productive performance of ewes
age of 10 months) were divided into three equal groups Birth weight of the neonates were recorded at the day of
(n=6) according to the body weight; control (C), AS50% birth and their weaning weights were recorded when
and AS100% to study the reproductive and productive lambs reached the age of 4 months. Daily weight gain of
performance. Ewe-lambs were fed on a commercial lambs was calculated using weekly body weight for each
concentrate feed mixture, clover hay and acacia leaves lamb.
according to NRC [6]. The acacia leaves were collected Milk production was measured weekly postpartum and
and offered fresh daily. The Alexandria University for seventeen weeks (119 day). Daily milk yield for each
guideline for the ethics and use of experimental animals ewe was performed using weigh suckle-weigh technique
was approved. [8]. Lambs were separated from their dams at 14:00 hr the
Reproductive performance of ewe-lambs day before milking and in the day after. The lambs were
This study was started two months before mating and weighed at 6:00 hr and left to suckle their dams till
continued till lambing. The control ewe-lambs were fed satisfactions. They were weighed again and kept in closed
on a commercial concentrates feed mixture and clover hay pens till next milking at 14:00 hr. In the meantime their
(1.0 kg/h/d). The AS50% animals were fed a commercial dams were striped to estimate the stripping milk. The
concentrate feed mixture and clover hay (0.5 kg/h/d) and same procedure was followed again at 14:00 hr in the
acacia leaves (1.0 kg/h/d). The AS100% animals were fed a same day. The daily milk yield was calculated by
commercial concentrate feed mixture and acacia leaves summing the weight of suckled milk (differences between
(2.0 kg/h/d). lamb's weight before and after suckling) and the weight of
Ewe-lambs were fed the commercial concentrate mixture striped milk in both morning and afternoon milking. Milk
in groups, hay and acacia leaves in about two equal parts production was evaluated using a graduated cylinder (5
at 8:00 and 14:00 hr daily. The ewe-lambs had free access mL). The amount of milk obtained was adjusted for 24 hr
to fresh water through the experimental period. Blood on weekly basis. Milk samples (100 mL) from individual
samples were collected every month from the jugular ewes were taken for proximate analysis using Milk
vein, before access to feed and water in test tube without Analyzer (Milko Tester Instruments Inc., Bulgaria).
anti-coagulant. The samples were centrifuged at 3000 rpm Milk urea nitrogen (MUN) was determined by adding 2
for 20 min to get the serum for biochemical parameters mL trichloroacetic acid (10%) to 5mL of milk sample for
assay. Total protein and albumin were assayed by protein precipitation and left for 30 min then the mixture
colorimetric kits (Stanbio, Boerne, Texas, USA). Glucose was centrifuged at 3000 rpm for 20 min in order to
was assayed using colorimetric kits (Futura System, separate the supernatant which was used to determine
Formello, Rome, Italy) and blood urea nitrogen (BUN), milk urea nitrogen by colorimetric method (Stanbio,
creatinine and cholesterol were assayed by colorimetric Boerne, Texas, USA) [7].
kits (BioSystems, Costa Brava, Barcelona, Spain). Blood samples were collected every month until the end
Globulin concentration was calculated as the difference of the experiment from the jugular vein, before access to
between total protein and albumin. feed and water in test tube without anti-coagulant to get
After two months of the beginning of the experiment, all coagulated blood samples. The samples were centrifuged
ewe-lambs were injected with prostaglandin PGF2 (0.5 at 3000 rpm for 20 min and sera were harvested for
ml of estroPLAN with the concentration of 125 g biochemical assays.
Cloprostenol; Parnell Technologies, Alexandria, New Samples analysis
South Wales, Australia). The animals were submitted to The dry matter (DM) of feed and feces were analyzed by
estrus detection twice daily using mature teaser ram at drying at 105C for 24 h. The AOAC (2006) analytical
7.00 and 19.00 hr for 3 days, ewe-lambs which were procedures were used for the organic matter (OM)
mounted by teaser rams, were mated by fertile rams. The determination (No. 968.08) by ashing at 600 C for 2 h, N
ewe-lambs which was insensitive to the effects of first estimation (No. 988.05) by a Kjeldahl technique and ether

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International Journal of Environment, Agriculture and Biotechnology (IJEAB) Vol-2, Issue-6, Nov-Dec- 2017
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extract. Neutral detergent fiber (NDF) and acid detergent and their interaction on different variables that were tested
fiber (ADF) were determined according to the procedure in a repeated measurement design as follows: Yijk = + Ti
of Van Soest et al. [9] without sodium sulfite or + Pj + (TP)ij +Akt + eijk; Where: is overall mean, Ti is a
amylase and expressed with residual ash. Concentrations fixed effect of the treatment (i=1 to 2), Pj is a fixed effect
of NH3-N and total volatile fatty acid (VFA) were of the time (j=1 to 8), TPij is an interaction between
determined according to Preston [10] and Warner [11], treatment and time, Akt is random effect of the animal
respectively. (within treatment) and eijk is random error assumed to be
Total phenols (TP) were determined with the Folin- independent by and normally distributed with mean = 0
Ciocalteau reagent according to Makkar [12]. Total and variance = 2. The reproductive traits results were
tannins (TT) were determined as the difference in TP compared among studied groups using Chi-square.
before and after treatment with insoluble polyvinyl RESULTS
polypyrrolidone (PVPP) [13]. Condensed tannins (CT) Experiment 1.
were measured by the HCl- butanol method according to Data of the effect of partial (50%) and total (100%)
Makkar (2003). TP and TT were expressed as tannic acid replacements of clover hay by acacia leaves on dry matter
equivalents and condensed tannins were expressed as intake (DMI) and apparent total tract nutrient digestibility
leucocyanidin equivalent [12]. of Barky sheep are shown in Table (2). There was a linear
Statistical analysis reduction (P<0.01) in DMI by increasing acacia leaves
First experiment data were subjected to analysis of compared to the control. Percentages of reductions in
variance (ANOVA) using the general linear model DMI due to partial (50) and total (100%) replacement by
procedure (GLM) of SAS software package [14]. The acacia leaves were 9.2 and 28.5%, respectively compared
used model was: Yij= + Fi + eij where: is the overall to the control. The apparent total tract digestion
mean, Fi is the treatment effect, eij is the random error coefficients of dry matter (DM), organic matter (OM),
term. The significant differences between individual crude protein (CP), natural detergent fiber (NDF) and acid
means were identified by using Tukey test. detergent fiber (ADF) were significantly lower (P<0.01)
Second experiment data were analyzed using GLM of in treated than control animals.
SAS [14]. The model included effect of treatment, time,

Table.2: Effect of partial and total replacement of clover hay by acacia leaves on apparent total tract nutrients
digestibility in Barky ewe lambs
Acacia
Item Control
AS50% AS100% P Value
Intake (g/h/d)
Clover hay 900.25.6 454.65.6 0 -
Acacia leaves 0 314.720.5 462.825.1 -
CFM 634.16.9 622.86.9 634.18.5 -
a b c
DMI g/d 1534.321.1 1392.221.1 1096. 925.9 <0.0001
Digestion coefficient (%)
DM 72.21.3a 67.01.3b 57.31.6c <0.0001
a b
OM 74.01.2 69.71.2 60.31.5c <0.0001
a b c
CP 68.91.6 55.71.6 38.02.0 <0.0001
NDF 73.01.5a 65.81.5b 52.61.9c <0.0001
ADF 68.82. 5a 57.82. 5b 35.53.0c <0.0001
Means in the same row with different superscripts significantly differ (P<0.05).
CFM : Concentrate feed mixture; DMI : Dry matter intake; DM : Dry matter; OM : Organic matter; CP :Crude protein; EE
:Ether extract; CF :Crude fiber; NDF : Neutral detergent fiber; ADF : Acid detergent fiber.

Table (3) presents data of the effects of acacia leaves (P<0.01) in nitrogen balance were found in animals given
replacement on nitrogen utilization, rumen NH3-N and partial (26%) and total (79.9%) acacia leaves compared
VFA concentration of Barky sheep. There were linear with control. There were no significant differences in the
(P<0.01) reductions in both nitrogen intake (NI) and concentrations of VFA and ammonia due to acacia
urinary nitrogen due to replacement compared to control. replacement.
In contrast, the fecal nitrogen increased (P<0.01) up to
48% with acacia replacement. Significant reductions
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International Journal of Environment, Agriculture and Biotechnology (IJEAB) Vol-2, Issue-6, Nov-Dec- 2017
http://dx.doi.org/10.22161/ijeab/2.6.9 ISSN: 2456-1878

Table.3: Effect of partial and total replacement of clover hay by acacia leaves on nitrogen utilization, rumen
NH3-N and VFA concentration in Barky sheep
Acacia
Item, g/d Control P Value
AS50% AS100%
N intake 32.300.26a 29.830.26b 24.100.26c <0.0001
b a
Fecal N 10.010.72 13.140.72 14.850.72a 0.0086
a ab b
Urinary N 9.900.76 7.520.76 6.740.76 0.0597
Nitrogen balance 12.390.51a 9.170.51b 2.510.51c <0.0001
VFA (meq/dl) 8.680.75 8.520.75 7.130.75 0.3066
NH3-N ( mg/dl) 14.350.64 13.060.78 13.410.64 0.4131
Means in the same row with different superscripts significantly differ (P<0.05).
VFA: Volatile fatty acids

Experiment 2.
The chemical composition of the total phenols (TP), total tannins (TT) and condensed tannins (CT) of acacia leaves
throughout the experimental period is presented in Figure 1.
30.00 Condnsed Tannins

Total Tannins
25.00
Total Phenol

20.00
g/kg DM

15.00

10.00

5.00

0.00

Month of the experiment


Fig.1: Total phenols, total tannins (eq-g tannic acid /kg DM) and condensed
tannins (eq-g leucocyanidin /kg DM) throughout the experimental period

The phenolic compounds like TP, TT and CT Table 4 shows data of the effects of partial and total
concentrations exhibited similar trends throughout the replacement of clover hay by acacia leaves on ewes
entire experimental period. They tended to decrease reproductive parameters. Results showed that number
gradually from February till June then increased slightly services per conception were similar (1.17) among
again to reach peaks in August and decreased thereafter groups. Likewise, conception rate have similar percentage
till December. Mean values of TP, TT and CT in acacia among groups, while fertility rate and lambing rate were
leaves were 100.4 g tannic acid kg-1 DM, 76.8 g tannic less in control than treated animals. The replacement of
acid kg-1DM and 36.1 g leucocyanidin kg-1 DM, acacia leaves has no significant effect on litter size.
respectively.

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International Journal of Environment, Agriculture and Biotechnology (IJEAB) Vol-2, Issue-6, Nov-Dec- 2017
http://dx.doi.org/10.22161/ijeab/2.6.9 ISSN: 2456-1878
Table.4: Effect of partial and total replacement of clover hay by acacia leaves on some reproduction traits of
Barky ewes
Acacia
Item Control
AS50% AS100%
Number services per conception 1.17 1.17 1.17
Conception rate, % 83.33 (5/6) 83.33(5/6) 83.33(5/6)
Fertility rate, % 66.66 (4/6) 83.33(5/6) 83.33(5/6)
lambing rate, % 66.66 (4/6) 83.33(5/6) 83.33(5/8)
litter size 1.0 1.0 1.0
Abortion rate, % 20 0 0

Effects of partial and total replacement of clover hay by serum albumin than the control. Serum cholesterol and
acacia leaves on blood biochemical parameters in creatinine were not affected during either stage by the
pregnant ewes are shown in Table (5). There were no inclusion of acacia leaves.
significant changes in all serum biochemical parameters Data of the effects of partial and total acacia replacement
during premating period between control and treated on BUN concentration at PGF2 injection and estrus and
animals. However, BUN declined (P<0.05) by 23% in the at 5, 10 and 15 days post estrus are presented in Table (6).
100% acacia-treated ewes compared to control. While The BUN was reduced (P<0.05) at day 10 after estrus,
during pregnancy, serum total protein, globulin, glucose, while there were no changes (P>0.05) in BUN
and BUN concentrations significantly decreased (P<0.05) concentrations at PGF2 injection, estrus, 5 and 15 days
by the increase of acacia leaves compared to the control. after estrus due to partial and total replacement by acacia
Contrariwise, results revealed that partial and total leaves.
replacements exhibited higher (P<0.05) concentration of

Table.6: Effect of partial and total replacement of clover hay by acacia leaves on blood urea nitrogen (BUN) concentrations
(mg/dl) at PGF2 injection and estrus and at days 5, 10 and 20 post estrus in ewes
Acacia
Item Control
AS50% AS100% P value
Day of PGF2 injection 21.081.1 23.581.1 23.821.1 0.1895
Day of heat 19.251.68 18.801.68 15.561.84 0.3107
Day 5 after heat 20.801.52 20.751.39 17.031.39 0.1317
a a b
Day 10 after heat 20.201.59 22.451.45 15.481.59 0.0199
Day 20 after heat 21.221.79 22.481.79 22.001.79 0.8822
Means in the same row with different superscripts significantly differ (P<0.05).

Table 7 illustrates effects of partial and total replacement significant differences (P>0.05) among groups. Typically,
of clover hay by acacia leaves on progesterone progesterone profile exhibited minimum values at the
concentration at PGF2 injection, estrus, 5, 10 and 20 days onset of heat and started to increase by time thereafter.
after estrus. Results indicated that there were no
Table.7: Effect of partial and total replacement of clover hay by acacia leaves on progesterone concentration
(ng/ml) in Barky ewes
Acacia
Item Control P value
AS50% AS100%
Day of PGF2 injection 4.900.68 3.900.68 4.600.68 0.5785
Day of heat 1.280.56 1.970.65 1.030.56 0.5598
Day 5 after heat 2.160.57 1.900.57 2.600.57 0.6888
Day 10 after heat 4.500.29 4.380.29 4.740.32 0.7094
Day 20 after heat 5.780.69 4.100.69 5.680.62 0.1959
Means in the same row with different superscripts significantly differ (P<0.05).
AS: Acacia saligna

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International Journal of Environment, Agriculture and Biotechnology (IJEAB) Vol-2, Issue-6, Nov-Dec- 2017
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Effects of partial and total replacement of clover hay by improvement due to partial substitution of acacia leaves
acacia leaves on milk yield, milk composition and milk was 11.8%, while the total substitution of acacia leaves
urea nitrogen (MUN) in lactating ewes are presented in decreased milk yield by 15.3% compared to the control.
Table 8. The results revealed that there is a linear Both AS50% and AS100% decreased (P<0.05) milk fat
reduction in DMI with increasing proportions of acacia percentage compared to control, while milk protein,
leaves in ewes diets. The AS50% group produced higher lactose, SNF and MUN were not affected (P>0.10) by
(P<0.05) milk yield than control and AS100%. The acacia replacement compared to control.

Table.8: Effect of partial and total replacement of clover hay by acacia leaves on Dry matter intake, milk yield, milk
composition and milk urea N (MUN) in lactating Barky ewes
Acacia P value
Item Control
AS50% AS100% Treatment Week TW
DMI g/d 1750 1685 1620 - - -
Milk yield g/d 509.922.0b 570.119.7a 431.722.0c <0.0001 <0.0001 0.9893
Fat % 5.470.22a 4.770.19b 4.460.22b 0.0033 <0.0001 0.7655
Protein % 3.880.07 3.850.06 4.010.07 0.2269 0.2671 0.1262
SNF % 10.700.19 10.710.18 11.060.19 0.3673 0.2772 0.1397
Lactose % 5.830.10 5.810.10 6.020.10 0.2907 0.3160 0.1106
MUN mg/dl 20.190.95 18.500.82 19.070.93 0.3768 0.1999 0.8854
Means in the same row with different superscripts significantly differ (P<0.05).
DMI: Dry Matter Intake, SNF: Solid Not Fat, MUN: Milk Urea Nitrogen.

Data of the effects of partial and total replacement of serum albumin, glucose and creatinine concentrations due
clover hay by acacia leaves on blood biochemical to the acacia leaves substitution at either level.
parameters in the lactating Barky ewes are shown in Contrariwise, serum cholesterol concentration was higher
Table 9. Serum total protein and globulins concentrations (P<0.05) in the AS100% than in C and AS50% ewes. BUN
declined (P<0.05) due to partial substitution of acacia concentrations were similar in C and AS50% (23.87 vs.
leaves (AS50%) but not in the AS100% ewes compared to 22.59 mg/dl) being greater (P<0.05) than that in AS100%
the control. There were no significant differences on (20.39 mg/dl) ewes.

Table.9: Effect of partial and total replacement of clover hay by acacia leaves on blood biochemical parameters in lactating
Barky ewes
Item Acacia P value
Control AS50% AS100% G T TxG

Total protein, (g/dl) 6.690.18a 5.950.16b 6.390.18ab 0.013 0.031 0.938


Albumin (g/dl) 4.100.07 3.950.07 4.010.07 0.298 <0.0001 0.443
a b
Globulin (g/dl) 2.590.17 1.990.16 2.380.17ab 0.043 0.002 0.985
Glucose (mg/dl) 45.871.76 44.351.64 42.171.88 0.300 <0.0001 0.0100
Creatinine (mg/dl) 0.990.05 1.030.04 1.010.05 0.830 0.051 0.275
ab b
Cholesterol (mg/dl) 63.192.47 56.672.30 66.162.47a 0.029 0.011 0.006
a a
BUN (mg/dl) 23.870.81 22.590.73 20.390.79b 0.014 <0.0001 0.089
Means in the same row with different superscripts significantly differ (P<0.05).
AS: Acacia saligna; BUN: Blood Urea Nitrogen

Table 10 presents data of the effects of acacia Similar trend was found with weaning weight as the
replacement on birth weight, weaning weight and average lowest weaning weight was found on lambs born of
daily gain of Barky lambs born of treated ewes. The AS100% ewes (15.59 kg) representing 22.7% weight loss,
highest birth weight was found in control-ewes lambs however AS50% - ewes lambs had less weaning weight by
(3.85 kg) which was comparable to those lambs born of 14.7% compared to C-ewes lambs. Although the partial
AS50% (3.09 kg) ewes. However, AS100% ewes lambs replacement of acacia leaves decreased the weaning
recorded the lowest (P<0.05) birth weight (2.97 kg). weight by 14.7% but the difference was not statistically

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International Journal of Environment, Agriculture and Biotechnology (IJEAB) Vol-2, Issue-6, Nov-Dec- 2017
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significant. Average daily gain of lambs decreased by non-significant difference between C and AS50%.
13.6 and 22.7% in AS50% and AS100%, respectively, with
Table.10: Effect of partial and total replacement of clover hay by acacia leaves on birth weight, weaning weight and
average daily gain (ADG) of Barky lambs
Acacia
Item Control
AS50% AS100% P value
Birth weight, kg 3.850.26a 3.090.20ab 2.970.26b 0.043
a ab b
Weaning weight, kg 20.171.21 17.200.94 15.591.21 0.033
ADG, g/d 145.779.19a 125.927.12ab 112.679.19b 0.042
Means in the same row with different superscripts significantly differ (P<0.05).
ADG: Average Daily Gain. Means in the same row with different superscripts significantly differ (P<0.05).

III. DISCUSSION volatilization and nitrous oxide emissions [24].


Feed intake and digestion Confirming these effects of CT-containing forages,
The reduction in DMI of sheep due to partial or total Powell et al. reported that the ratio of N excreted in feces
substitution of clover hay by acacia leaves hay might be and urine was highest for low-tannin and high-tannin
due to astringency, decreased palatability, which possibly birds foot trefoil treatment and lowest for the alfalfa
resulting in feeds avoidance and adverse effects on treatment [25]. Current study data revealed that the higher
digestion as reported previously [12, 15]. Moreover, fecal N excretion in the presence of active tannins was
Waghorn et al. [16] demonstrated that decreased ruminal matched by lower urinary N excretion. This shift can be
turnover and rate of digestion were more important than explained by the lower rumen degradability of
palatability in reducing intake of sheep fed diets nitrogenous compounds and is confirmed by effects of CT
containing high levels of CT. Other researchers suggested on rumen fluid ammonia and total N as previously stated
that high concentrations of tannins could reduce the intake [26, 27]. Volatile fatty acids production was not affected
in the following ways: (1) physical distension of the by acacia leaves replacement. The possible explanation
rumen, resulting in a decrease in dry matter digestion; (2) for this might be due to lack of effect of supplemented
hormonal response due to the binding of tannins to the gut tannins on rumen bacteria or to the adaptation of rumen
wall; (3) reduction of the diet palatability caused by its microorganisms to tannins [28].
astringency; and/or (4) binding of tannins to salivary and Reproductive performance
mucosal proteins [17, 18]. Conception rate was similar among treatments; however
The depression in apparent total tract digestibility of CP fertility and lambing rates were higher by about 25%
associated with replacement of clover hay by acacia when acacia leaves were replaced in ewes diets. These
leaves might have been due to the formation of complexes effects probably occur as a result of including tannins-rich
between tannins and dietary proteins and carbohydrates plants in ewes diets, which enhance the live body weight,
[19], as well as reducing rumen microbial proteolytic, body condition, and energy and protein intake and protein
ureolytic and cellulolytic enzyme activities, general absorption from the small intestine [29]. Moreover,
fermentative activities and cell multiplication [20]. increased plasma of essential amino acids principally
Tannins might also interfere with digestion by binding branched chain amino acids and plasma metabolic
microbial enzymes and this might explain why acacia- hormones especially insulin were found in tannin-fed
given animals in this study decreased apparent total tract animals [30]. Short periods of improved nutrient supply
digestibility of cell walls digestion [21]. Animals given before and during mating and reproduction have been
acacia in the current study tended to decrease N balance known to affect ovulation rate along with increased size
[22]. This is probably due to the presence of CT, high and number of follicle [31], reduce follicular atresia [32],
proportion of acid detergent insoluble nitrogen and high altered plasma gonadotrophin concentration [33] and
urinary N, which in turn was possibly attributed to an affect ovarian sensitivity to gonadotrophins [32]. A large
imbalance of high N relative to a low energy in the part of the dietary protein is hydrolyzed in the rumen to
rumen. The interaction of tannins with protein alters the ammonia, some of which is re-incorporated into microbial
partitioning of N within the sheep guts, shifting the route protein. Excess ammonia is absorbed from the rumen and
of excretion away from urine toward feces [3, 23]. This metabolized to urea in the liver, leading to increased
reduction in urinary N reduces volatile N losses after land plasma and uterine ammonia and urea concentrations [29]
application with dairy manure, which in turn reduces which may increase the number of early embryonic losses
environmental losses through nitrate leaching, NH3 [34]. In agreement with the present finding, subsequent

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International Journal of Environment, Agriculture and Biotechnology (IJEAB) Vol-2, Issue-6, Nov-Dec- 2017
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grazing experiments with sheep showed that CT in L. 75 g/day) [44]. When the level of acacia was doubled, the
corniculatus increased both ovulation rate and lambing beneficial effect of acacia disappeared since the average
percentage by 20-27% [29]. The improvement of the daily gain of lambs was 82 g/day. These findings raised
conception rate in the current study might be ascribed to the question as to whether the positive effect of 100 g
that BUN level was less than 20 mg/dL at the acacia is because of the interaction between acacia tannins
insemination day. The results of the BUN is confirmed by and soybean meal protein.
Butler who indicated that conception rate decreased when
serum urea nitrogen concentrations exceeded 20 mg/dL Blood metabolites
on the day of insemination and suggested that degradation Plant secondary compounds may affect blood parameters
of excessive amounts of dietary protein in the rumen by maintaining them [45], while others may decrease [46]
contributed to infertility [35]. or increase [47] plasma glucose concentration, or alter
Milk production and composition serum insulin concentration [48]. In agreement with our
Effects of tannins-rich feeds on milk fat and protein finding, Waghorn et al. and Ben Salem et al. reported that
composition varies markedly depending on the BUN was lower when sheep and goats were fed legumes
concentration of tannins present in the feeds. Condensed that contained tannins [44, 49]. However, reduced
tannins in high concentrations (e.g. total replacement by proteolysis in ewes receiving Acacia cyanophylla with
acacia) generally have adverse effects on animal concentrate could have been caused by effects of acacia
performance, while moderate concentrations (e.g. partial tannins on microbial proteolytic activity [50].
replacement by acacia) might have positive effects. The Additionally, others demonstrated that that BUN was also
negative effects of tannin-diets on milk yield and fat higher in the Bermuda grass hay-based ration compared
content might be due the reduction of feed intake, with the Lespedeza cuneata forage (23.1 mg CT/mg
decreased rate of digestion and development of soluble protein) in goats [51, 52].
conditioned aversion [36, 37]. Wang et al. reported that Solaiman et al. reported that BUN, albumin, creatinine,
tannins from Lotus corniculatus, which contained triglycerides and glucose were unaffected in the diets of
moderate amounts of CT (44.5 g/kg DM), fed to lactating goats consuming different levels of the CT containing
ewes increased milk yield, lactose and protein content and forage of sericea lespedeza (Lespedeza cuneata) [53],
decreased milk fat as found in the present study[38]. One however our data indicated elevation (P<0.05) of serum
reason for these effects could be an increase in albumin due to partial substitution of clover hay by acacia
metabolizable protein supply from the protein binding leaves, while total substitution decreased (P<0.05) serum
action of CT because effects of tannins on ruminant albumin. It is well known that serum albumin is a bio-
productivity depend on the quality and quantity of dietary indicator of the nutritional status of the animal and it
protein [28]. Protein protection by tannins from microbial decreases when animals are diseased. The glucose values
degradation in the rumen resulted in an increase of milk ranged from 61.2 to 65.6 mg/dL in the current study,
production in dairy cows [39], dairy goats [40] and sheep which were similar to previous studies [51, 53].
[41]. Contrariwise, reported no improvement in ewes Silanikove et al. demonstrated that the blood metabolic
milk yield when acacia leaves were supplemented into profile was examined in non-lactating and non-pregnant
their diets at 100 or 200g/day [42]. goats consuming Quercus calliprinos (oak), Pistacia
Parallel to our finding, Maamouri et al. indicated that lentiscus (pistacia) and Cerafonia siliqua (carob) leaves
milk protein content hasnt been affected significantly in [54]. Overall view of the data of this study revealed that
acacia-treated ewes [42]. On the contrary, these results the controversy in the literature with the current study
disagree with those of Wang et al. for dairy sheep [38] implying the ability of tannins to modulate the rumen
and Woodward et al. (1999) for dairy cows [39] who fermentation, nutrients utilization and performance
found increased milk protein content when tannin was efficiency of ruminants is probably due to the great
supplemented to the diet. The CT-containing plants can diversity in the structural features, ruminants species, and
protect dietary protein against degradation in the rumen consequently in the reactivity of these plant secondary
and increase N utilization, resulting in a reduction in compounds. The dose-dependent effect of tannins and
MUN concentration and nitrogenous waste excretion and proportions of rumen degradable protein are another
improved nutritional status of the animal. However, Ben major issues because of the difficulty in selecting
Salem et al. found that supplementing lambs fed on concentrations to positively affect a particular parameter
spineless cactus pads with small amount (i.e. 100 g/day) without conferring a negative response on others (e.g., in
of air-dried acacia and 200 g soybean meal significantly overall diet utilization).
increased their growth rate compared to those receiving
the same diet but without acacia leaves (102 g/day versus IV. CONCLUSION

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International Journal of Environment, Agriculture and Biotechnology (IJEAB) Vol-2, Issue-6, Nov-Dec- 2017
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Partial (up to 50% of the forage) replacement of clover [10] Preston, T.R. (1995). Biological and chemical
hay by acacia leaves could be implemented in sheep analytical methods. In: T.R. Preston (Ed.) Tropical
feeding without detrimental effects on the utilization of Animal Feeding: A Manual for Research
nutrients, fermentation characteristics, reproductive and Workers. FAO, Rome, pp:191-264 (Chap. 9).
productive performance. [11] Warner, A.C.I. (1964). Production of volatile fatty
acids in the rumen. Methods of measurements. Nutr.
ACKNOWLEDGMENTS Abs. Rev. 34, 339.
The authors express their sincere gratitude to the farm [12] Makkar, H.P.S. (2003). Effects and fate of tannins in
crew for their endless help. ruminant animals, adaptation to tannins, and
strategies to overcome detrimental effects of feeding
CONFLICT OF INTEREST tannin-rich feeds. Small Rum. Res. 49, 241-256.
The authors declare that they have no competing interests. [13] Makkar, H.P.S., Blummel, M., Borowy, N.K.,
Becker, K. (1993). Gravimetric determination of
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Table.5: Effect of partial and total replacement of clover hay by acacia leaves on blood biochemical parameters in the
premating and pregnant Barky ewe-lambs
Item Premating period Pregnancy period P value
Control Control G T Tx
AS50% AS100% AS50% AS100%
G
Total protein 8.230.32 7.700.16 7.130.16 6.970.18 0.0002 0.509
8.560.32 8.290.34 A B B 0.004
(g/dl)
Albumin 4.250.14 4.120.13 4.320.05 4.580.05 4.430.05 0.001
4.080.13 B A A 0.002 0.002
(g/dl)
Globulin(g/d 3.380.17 2.540.17 2.560.19 <0.00
4.500.34 4.050.35 4.120.34 A B B 0.0006 0.524
l) 01
Glucose(mg/ 55.594.1 61.384.1 68.912.8 57.012.8 62.453.3
61.964.35 0.020 0.0009 0.089
dl) 5 5 6A 6B 2B
Creatinine
0.980.06 0.890.06 1.030.6 1.030.04 0.990.04 0.960.05 0.604 0.001 0.311
(mg/dl)
Cholesterol 34.932.5 41.612.5 47.811.7 47.121.7 48.362.0 <0.00 0.031
37.032.69 0.937
(mg/dl) 7 7 7 7 5 01 6
<0.00 0.035
BUN 26.101.3 22.681.38 19.991.3 20.690.6 20.130.6 13.130.7 <0.00
0 2
(mg/dl) 1A AB
1B 5A 5A 5B 01
1
AS: Acacia saligna; BUN: Blood Urea Nitrogen.

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