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STUDY THE IMPACT OF MONENSIN ,VITAMIN E AND DATE
PALM SEEDS EXTRACT
ON CERTAIN ARABI RAMS SPERM PARAMETERS.
Khairi,G.Alrikabi*, Kareem.H.Muhanna* , Ziad ,G.kushan**
*Technical Institute of Shatra,Thi-Qar-Iraq
**Ministry of Science and Technology,Thi-Qar, Iraq
(Received 3 February 2020,Accepted 9 September 2020)
Key words: Monensin , vitamin E ,dates palm seeds.
corresponding author: [email protected]
ABSTRACT
The aim of this study is to assess the impact of monensin, vitamin E and dates palm
seeds extract(DPSE) on some parameters of Arabi rams sperm .Twenty rams were
used in this study, randomly distributed in 4 groups, each group contains 5 animals.
which treated daily as the following : control (without addition), T1(50 ml of
(DPSE)/head), T2 (50 mg vit E / head ) and T3 (200 mg monensin /head).The
experiment continued for two months. Semen collected weekly by artificial vagina.
After collecting the semen the sperm parameters were evaluated in addition to
measurement the testosterone level and some antioxidative enzymes and oxidative
marker(MDA). The results showed that T1 and T2 exhibited enhancement in semen
volume,sperm concentration, individual motility and testosterone levels
while MDA levels are reduced and elevation of GPx and SOD as compared to control
and T3. It was concluded that DPSE and vit E have advanced effect on sperm
parameters of Arabi rams.
INTRODUCTION
Reproductive efficiency is one of the main pillars of the success of animal
breeding projects which enhanced through dissemination of high value genetic factors
to increase productivity and improve income of breeders(1).
It is possible to increase reproductive efficiency by manipulation with the
component of the the ration or supporting it with some additives. Researches have
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revealed that date seed contains high nutritional value, especially in term of their
content of fibers and antioxidant (2).
Date palm seed comprises various chemical compounds such as saturated
fatty acids (stearic and palmitic acid) , un saturated fatty acids( linoleic,oleic acids)
which could inhibit the 5 - α reductase enzyme , Zinc(Zn) ,Calcium(Ca)
,potassium(K)(3) , fat ,protein , , dietary fiber of ash . Also date seed contains high
level of phenolics , antioxidant(4), ferulic, sinapic, p- coumaric , and acids,
procyanidins and flavonoids (5; 6) and lipid, which is either in wax , fat or oil
form(7).
Vit E is another additives which is called as anti-sterility since it is required for
female reproductive system to function normally. (8), and (9), indicated that vitE is a
main chain - breaking anti-oxidant in sperm membrane because it can instantly
scavenge free radical such as hydrogen peroxide(H2O2), superoxide anion(•O2-), and
hydroxyl radicals(OH•) . Due to its lipid solubility, vitamin E is the main line of
protection from peroxidation of the polyunsaturated fatty acid of phospholipids in
sperm membrane (10). Monensin belongs to the ionophor group, which increases the
plasma membrane permeability of bacteria, causing them to burst and convert to
beneficial microbial protein. monensin are reducing the number of methanogenic
bacteria (11) which reduce the energy lost due to methane production. (12) and (13)
mention that monensen should cause an influx of Ca++, there being abundant Ca++
existent to induce capacitation.
MATERIALS AND METHODS
Animals and Collection of semen
These experiments were done in local field in Al-Nasir city, (Thi-qar province)
from 5-1-2018 to 5-3-2018 In this study 20 Arabi rams with average 3.5- 4 years old
and 56 ± 5 kg body weight were chosen and were held in pens with semi-slatted
floorings. Animals were fed a diet based on (14) recommended based on 60:40
forage to concentrate ad libitum with free access to water.The animals were divided
randomly into 4 groups (5 rams for each group) and they were treated daily as follow:
control(without addition), T1(50 ml of date palm seeds extract/head), T2 (50 mg vit
E/head ) and T3 (200 mg monensin /head). After completion of the preliminary period
(two weeks) for adaptation on new diet and training for semen collection, 10
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ejaculates (each six days) were collected from each ram every six days by using
artificial vagina as described by (15).
Preparation of Date Palm Seed Extract
Date fruits were obtained from the Qalat Sekar orchards in Thi-Qar province; The
seeds were picked, rinsed to dry and roasted well afterwards. The dry pits were
crushed into a fine powder and added distill water to make a 50 gm/L mixture which
was boiled till it becomes brownish in colour and finally filtered.(16).
Semen Processing
After the beginning of the treatments ,semen was collected with an artificial
vagina every six days , equipped with graded test tubes, sited in a thermo flask at
37°C, and transferred to the laboratory for assessment. (volume of semen, PH, count
of sperm, mass and individual motility, viability and sperm abnormality.
Table(1 ) Chemical analysis of diets and their content of metabolized energy
Composition ration
Dry matter 92.05
Crude protein 14.69
Ether extract 5.52
Crude fiber 8.37
Nitrogen free extract 59.57
Organic matter 88.15
ash 3.90
Metabolized energy mega joules /kg dry
matter
12.232
Metabolized energy based on values for ruminants(14)
Table(2) Primary feed materials and additives used in experimental diets
diet control T1 T2 T3
barley 31 31 31 31
Soybean 15 15 15 15
Wheat flour 10 10 10 10
bran 25 25 25 25
Ground corn cobs 15 15 15 15
Minerals and vitamins 3 3 3 3
salt 1 1 1 1
total 100 100 100 100
additives
DPSE ml/head 50
Vit E mg/head 50
Monensin mg/head 200
based on values for ruminants(14)
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Measurement of testicular Malondialdehyde (MDA Concentration):
Testicular lipid peroxidation product(MDA) was done by the way of Yashkochi
and Master (17).Reaction of MDA with thio-barbituric acid(TBA) in an acidic media
give a colored TBA_complex estimated by spectrophotometer at 520_535 nm in
contrast to blank and the value of MDA were stated as n mole MDA/mL semen.
Estimation the activity of testicular Super-oxide Dismutase :
Superoxide dismutase activity(SOD) was assessed rendering to Giannopolitis and
Ries(18). The optical absorbance was estimated at waves length (560)nm against
blank reagents. SOD = Reading (absorbances) of (SOD)/ mL semen.
Measurement activity of testicular Glutathione peroxidase:
Activity of glutathione peroxidase in semen was determined
spectrophotometrically using Ransel reagents from Randox Laboratory Ltd.
(Crumlen,UK). Glutathione is oxidized by cumene hydroxyperoxide and is directly
reduced to glutathione by glutathione reductase with a simultaneous oxidation of
NADPH to NADP+. The activities are stated as unit(U) /mL semen.
Measurement of CAT activity:
The method described by (19) was used for the determination of CAT activity in
the semen samples.
Statistical analysis:
Data were analyzed using SPSS-Version 19). Two ways ANOVA and least
significant differences (LSD) were performed to assess significant difference among
means P≤0.05.
RESULTS
The results in table (3) showed that DPSE and Vit E treatment presented high
significant difference(P≤0.05) as compared to control and monensin treatment in
semen volume (1.40±0.22 and 1.30±0.26 vs. 1.00±0.34 and 1.00±0.30 respectively)
and concentration (3.4±0.33 and 3.2±0.36 vs. 2.8±0.23 and 2.9±0.18 respectively)
DPSE exhibited higher significant difference(P≤0.05) in an individual
motility(88±3.14)as compared to control group(80±2.22) and non-significant
difference as compared to vit E and monensin group (85±3.24 and 82±2.18)
respectively. The results also revealed that DPSE and Vit E groups showed lower
significant difference(P≤0.05) in an abnormality compared to control and monensin
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groups (6.5±0.76 and 6.8±1.16 vs. 8±1.62 and 7.8±0.98 respectively) while there was
non-significant difference among treatment in PH, mass motility and viability.
Table (3) In vivo Effect of DPSE ,Vitamin E and Monensin on Some Sperm
Parameters of Arabi Rams(means±SE).
parameters control DPSE vitE monensin LSD
volume 1.00±0.34
b
1.40±0.22
a
1.30±0.26
a
1.00±0.30
b
0.30
PH 6.8±1.2
a
7.0±1.3
a
7.0±0.8
a
6.9±0.70
a
NS
Concentration
mil/ml
2.8±0.23
b
3.4±0.33
a
3.2±0.36
a
2.9±0.18
b
0.3
Mass motility% 90±4.5
a
95±4.4
a
92±6.4
a
92±7.20
a
NS
Individual
motility%
80±2.22
b
88±3.14
a
85±3.24
ab
82±2.18
b
6.0
Viability%
90±4.23
a
95±3.19
a
95±6.30
a
92±4.05
a
NS
Abnormality% 8.0±1.62
a
6.5±0.76
b
6.8±1.16
b
7.8±0.98
a
1.0
Different small letters vertically denotes significant (P<0.05) between groups
Results in Table (4) revealed that DPSE and Vitamin E were significantly (P <
0.05)higher than control and monensin group in level of testosterone (16.5±1.24 and
16.0±2.20 vs. 12.3±1.13 and 13.1±2.18 respectively), GPx(3.97±0.88 and 3.76±0.67
vs. 1.97±0.97 and 2.60±0.47 respectively) and SOD (20.65±1.66 and 18.92±1.16 vs.
16.15±2.60 and 17.32±1.24 respectively) while there was a low significant
difference(P≤0.05) in the level of MDA (3.22±0.45 and 3.02±0.32 vs. 5.17±0.78 and
4.32±1.13 respectively ) , Non-significant differences among treatments in the level of
CAT.
Table(4) In vitro Effect of DPSE ,Vitamin E and Monensin on Some Sperm
Parameters of Arabi Rams(means±SE).
parameters control DPSE Vit E monensin LSD
Testosterone
ng/mL
12.3±1.13
b
16.5±1.24
a
16±2.20
a
13.1±2.18
b
3.7
MDA(nomL/
mL)
5.17±0.78
a
3.22±0.45
b
3.02±0.32
b
4.32±1.13
a
1.1
GPx(U/mL) 1.97±0.97
b
3.97±0.88
a
3.76±0.67
a
2.60±0.47
b
1.16
SOD(U/mL) 16.15±2.60
b
20.65±1.66a 18.92±1.16a 17.32±1.24
b
1.6
CAT(U/mL)
4.11±0.40
a
3.54±0.75
a
3.62±1.42
a
4.50±1.04
a
NS
Different small letters vertically denotes significant (P<0.05) between groups
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DISCUSSION
Daily oral treatment with pits extract of DPSE and vit E cause a significant
increase in most sperm characteristics such as volume, sperm concentration and
individual motility with reduction of abnormalities of Arabi rams spermatozoa. This
findings are confirmed by (20) who established that the relative estimation between
control and treated groups show that administration of date palm pits suspension
enhanced sperm count, motility, morphology . Improvement in sperm parameters in
DPSE treatment may be referred to its content of of estrogenic constituents as
gonad_stimulating compound that increase male fertility (21) as well, date seed
contains a substantial amount of food ingredients such as proteins and mineral that
improve sperm quality (22).
These findings of vit E treatment were in agreement with the observations of
(23) who studied the impact of dietary vit E on sperm motility and kinetic traits of
ram sperm, the positive effect of vitamin E is related to its biological functions that
prevent lipoprotein oxidation of sperm membrane(24). vitamin E keeps and decreases
gaps in the helix of mitochondria of mid piece of spermatid during late
spermatogenesis that may be responsible for improved motility (25). (26) found that
vitamin E regulates phospholipase A2 activity that plays a pivotal role in stabilization
of viable sperm by interaction with phospholipids. Feeding vitamin E may elicit direct
stimulation of steroidogenic enzyme , modification of cAMP second_messenger
function and / or interfering with cell membranes properties that could lead to
maintain the conformation of cell membrane (27).
Slight enhancement of the semen characteristics in monensin treatment was in
agreement with that obtained by (28). A possible mechanisms involve in the
monensin_mediated rise in LH liberation was proposed to be an influence at the level
of the Golgi apparatus of endocrine cell. In vitro researches with pituitary fragments
of perfused rat, proposed capability of monensin on redirecting hormone production
toward secretion more than storage(29).
The daily oral dealing with pit of date palms produced a noteworthy rise in the
level of testosterone in blood serum of Arabi rams, this agrees with (30)who stated
that a date palm pollen suspensions increase the level of estradiol and testosterone in
plasma which are found at high concentrations in seminal plasma and rat testis.Also,
(21) detected that DPSE increase the sperm concentration in guinea pigs and increase
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the level of testosterone in rats. DPSE have been involved in animals ration as growth
promoter, the effect has been ascribed to an enhancement in the plasma level of
testosterone(31). Intra_testicular testosterone levels designates that date palm
suspension changed leydig cell steroidogenesis. Though, the particular way of this
effect is unknown (32).
Treatment with date palm seeds and vit E cause reduction in the level of MDA
and increase GPx and SOD. (33) showed that date palm comprises estradiol and
flavonoids and other antioxidant constituents that improve the quality of sperm. The
therapeutic use of DPSE is an ancient practice. It has been suggested that its
beneficial therapeutic effects referred to their antioxidant constituents. Dates have
phenolic compounds mainly(cinnamic acids) and flavonoids(flavones, flavonols, and
flavanones) that provided antioxidant activities(34).)Our result was in accordance to
that obtained by (35). Oral administration of vitamin E has valuable effects on the
male reproductive system. The beneficial effects were apparent in the form of
increase of semen quality, antioxidants status and testosterone in mammals. Vit.E is a
lipid-soluble antioxidant that neutralize free radical and keeps cellular membrane
against O2 free radicals. It furthermore avoids lipid peroxidation and therefore
enhance functions of other antioxidants(36). It was concluded that DPS and vit E
enhanced sperm parameters of Arabi rams due to their antioxidant effect.
دراسة تأثیر الموننسین وفیتامین ھ وخلاصة نوى التمر فی بعص معاییر النطف للخراف
العرابیة
خیری غرکان عوید الرکابی*، کریم حمادی مھنى* ، زیاد کاطع کوشان**
* المعھد التقنی،الشطرة، ذی قار ، العراق.
** وزارة العلوم والتکنولوجیا، ذی قار ، العراق.
الخلاصة
اجریت ھذه الدراسة لتقییم تأثیر المونینسین وفیتامین ھ وخلاصة بذور النخیل فی بعض معاییر النطف
لدى الکباش العربیة. استخدم عشرون کبشاً عربیاً فی ھذه الدراسة ، وزعت عشوائیاً الى ٤ مجموعات ، کل
مجموعة تحتوی على ٥ حیوانات عوملت یومیا على النحو التالی: السیطرة(بدون اضافة)، المعاملة الاولى ( ٥٠
مل خلاصة بذور النخیل/رأس)،المعاملة الثانیة( ٥٠ ملغم فیتامین ھ/رأس) والمعاملة الثالثة( ٢٠٠ ملغم
موننسین/رأس). جمع السائل المنوی اسبوعیا باستخدام المھبل الاصطناعی. بعد جمع السائل المنوی تم تقییم
معاییر النطف وکذلک تقدیر مستوى ھرمون التستسترون وبعض انزیمات الاکسدة. تبین من النتائج ان المعاملتین
الاولى والثانیة اظھرتا تحسنا معنویا فی حجم السائل المنوی وترکیز الحیوانات المنویة والحرکة الفردیة
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ومستوى ھرمون التستسترون بینما کان ھناک انخفاض فی مستوى المالونیلدھاید وارتفاع مستوى الکلتوثایون
بیروکسیدیز والسوبر اوکساید دیسمیوتیز. یستنتج مما سبق بان خلاصة بذور النخیل وفیتامین ھ لھما تأثیر
محسن فی صفات نطف الکباش العرابیة.
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