Vol. 1 Issue 5, pp: (99-107), August 2016. Available online at: http://www.prudentjournals.org/IRJAFS
International Research Journal of Agricultural and Food Sciences
Article Number: PRJA10909582
Copyright ©2016
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Author(s) agree that this article remain permanently open access under the terms of the Creative Commons
Attribution 4.0 International License.
Full Length Research Paper
Effect of gonadotrophin (Pergonal (R)) on semen
characteristics, hormonal profile and biochemical
constituents of the seminal plasma of mature West
African Dwarf rams.
Uche Nfa Egu1 and Ugochukwu Hart Ukpabi 1*
1
Department of Animal Science and Fisheries, Abia State University,
P.M.B. 7010, Umuahia Campus, Nigeria.
*Corresponding author. Email: [email protected]
Received 28 February, 2016; Accepted 27 July, 2016.
ABSTRACT
Twenty–four West African Dwarf rams aged 2.0 – 2.5 years were randomly distributed into 4 groups
of 6 animals per group and used to determine the effect of Pergonal ® on semen characteristics,
hormonal profile and biochemical constituents of seminal plasma. The doses were 0.00, 0.11, 0.22
and 0.33ml Pergonal (R) represented as T1, T2, T3 and T4 respectively. The group that received 0.00ml
Pergonal(R) served as the control. All the treatments were administered within 3days by intramuscular
injection. Rams that received 0.22ml of Pergonal had the highest percentage of live sperm cells
(80.20%) and normal sperm cells (77.30%) and the lowest percentage of dead sperm cells (19.80%).
The concentration of testosterone in the seminal plasma decreased from 0.95ng/ml in the control to
0.60ng/ml in T4 (0.33ml). There were significant differences (P<0.05) among the treatment groups in
urea, glycerophosphocholine, ascorbic acid, sodium, potassium, bicarbonate and fructose levels in
the seminal plasma. Pergonal had no significant (P>0.05) effect among the treatment groups in
luteinizing hormone level in the seminal plasma. The results of this study showed that up to 0.22ml
Pergonal may be used to enhance semen quality and biochemical constituents of the seminal plasma
of West African Dwarf rams.
Key words: Pergonal, Semen quality, Hormones, Seminal Plasma Constituents, WAD rams.
INTRODUCTION
The primary aim of induction of spermatogenesis
is to improve semen quality (Abu et al., 2006;
Ameh, 2004). Spermatogenesis involves the use
of follicle stimulating (FSH) and luteinizing
hormones (LH) (Abu et al., 2006). Most of these
preparations of FSH and LH are very expensive
perhaps because of the brand names. Some of
them require cold chain storage and often
deteriorate because of inadequate storage and
handling (Herbert et al., 2000). There is therefore
the need to examine some generic preparations
that could induce spermatogenesis in the
animals but at the same time are cheap, readily
available and easily managed under developing
countries conditions.
Pergonal® a fertility drug (Ferring Labs, USA)
also known as Humegon or Mentrophin and with
similar constituents as Plusset ® is a
gonadotrophin lyophilized in vials containing a
mixture of gonadotrophin consisting of FSH and
LH in a ratio of 1:1 (Dixon and Hopkins, 1996).
Follicle stimulating hormone and LH present in
Pergonal play vital roles in the initiation of
100
Int. Res. J. Agric. Food Sci.
spermatogenesis (Abu et al., 2006). Luteinizing
hormone stimulates the interstitial cells of Leydig
to produce androgens, mainly testosterone. The
androgens are secreted into the blood stream,
where they cause the development of secondary
sex characteristics of the male and the
development and maintenance of male
reproductive tract (Brackett, 2005). The
androgens suppress gonadotrophin releasing
hormone (GnRH), LH and FSH secretion by
negative feedback on the pituitary and
hypothalamus. Testosterone is also secreted
into the seminiferous tubule, where it is
necessary
for
the
maintenance
of
spermatogenesis (Garner and Hafez, 2000).
Follicle stimulating hormone reacts with
receptors on the Sertoli cells to cause production
of androgen-binding protein (ABP), conversion of
testosterone to dihydrotestosterone
and
estrogen, stimulation of spermatogenesis,
completion of sperm release (Spermiation) and
secretion of inhibitin (Osinowo, 2006). The
inhibitin secreted into the blood stream has a
negative feedback effect on the FSH but not on
the secretion of LH.
It has not been determined if the administration
of hormone preparation for spermatogenesis
would induce any side effects on the hormonal
profile and seminal plasma constituents of the
West African Dwarf (WAD) rams. This study was
therefore carried out to determine the effects of
Pergonal administration on semen quality,
hormonal
profile
and
seminal
plasma
constituents of West African Dwarf rams.
MATERIALS AND METHODS
Experimental Animals and Their Management:
Twenty-four healthy, sexually matured West Africa
Dwarf rams aged 2 - 2.5 years were used for this
study. The animals were purchased from the local
markets and housed in clean pens constructed in
such a way that the rams could come outside
during the day for access to sunlight and forage.
The animals were dewormed and routine
inspection for cleanliness was carried. Freshly cut
forage (Panicum maximum, Aspilia africana and
Pennisetum purpureum) was supplied as basal
diet and a concentrate ration of growers’ mash was
used as supplement. The animals were fed twice
daily, in the morning and evening. Salt lick was
provided as mineral supplement. Water was given
ad libitum to the animals.
Experimental Design and Drug Administration
Twenty-four WAD rams were divided into 4
treatment groups consisting of 6 rams per group
with one ram per replicate in a Completely
Randomized Design (CRD). These groups were
assigned to 4 levels of Pergonal as treatments. The
levels of Pergonal were 0.00, 0.11, 0.22 and
0.33ml Pergonal® represented as T1 T2, T3 and T4
respectively. T1, which contained no Pergonal
served as the control.
Pergonal was supplied in 24 vials, each vial
containing FSH 75 I.U and LH 75 I.U. The content
of the first vial was dissolved in 1 ml of physiological
saline solution immediately prior to use, resulting in
a solution of Pergonal-follicle stimulating hormone
(75 I.U) plus Pergonal-luteinizing hormone (75 I.U)
per ml.
Group T1: Each ram received 1.00ml of
physiological saline for 3 days.
Group T2: Each ram received 0.03ml of Pergonal
on the first day. Second day, the group received
0.04ml of Pergonal while on the 3rd day, the group
also received 0.04ml of Pergonal giving a total of
0.11ml Pergonal injections within 3 days.
Group T3: Each ram received 0.06ml of Pergonal
on the first day. Second day, the group received
0.07ml of Pergonal. While on the 3rd day, the
group received 0.09ml of Pergonal, giving a total of
0.22ml of Pergonal injection within 3 days.
Group T4: Each ram received 0.11ml of Pergonal
on the first day. Second day, the group received
0.11ml, while on the third day, the group also
received 0.11ml of Pergonal, giving a total of
0.33ml of Pergonal injections within 3 days. All
treatments were administered intramuscularly on
the hind leg (thigh) of each ram using a one ml
syringe with 0.01 ml graduation.
The doses were based on the manufacturer’s
prescription and procedure (Wierschoz et al.,
1996)
Semen Collection and Evaluation
Semen collection was done by Electro-ejaculation
method (Noakes et al., 2001) after one week of
Pergonal administration and continued at 2 weeks
interval for 9 weeks. The semen was collected
between 8.00 A.M and 9.00 A.M. A transparent
Egu and Ukpabi 101
graduated tube immersed in a protective jacket
containing water at 37oC, with a funnel was used to
collect the semen. The animal’s hind limbs were
held up by an assistant, with shoulder and fore
limbs of the animal held tightly between the two
legs of the assistant. The Vaseline lubricated probe
was inserted gently into the rectum. The rhythmic
stimulation of the ampullae and sacral nerve plexus
caused erection and subsequently ejaculation
within few minutes. Two ejaculates were collected
from each ram and evaluated and the means used
as the results for semen characteristics.
Semen evaluation was done as promptly as
possible post collection as described by
Rodriguez-Martinez and Barth (2007) for
qualitative and quantitative parameters such as
semen volume, sperm concentration, dead sperm
percentage, live sperm percentage, sperm motility
and pH.
Semen Volume: The volume of semen collected
was measured in ml using the graduated collection
tube.
Mass Motility: Using a sterile dropping pipette, a
drop of semen was placed on a warm slide while
the slide was observed under x10 magnification of
a light microscope. The warm slide was
obtained by placing the slide on a warm
chamber for 2 minutes. The motility estimate
was done by taking estimate of sperm waves
from three different apexes of the angle and
finding the average score (Umesiobi and
Iloeje, 1999; Rodriguez-Martinez and Barth,
2007).
Individual Motility: Using a dropping pipette, a
drop of semen was placed on the warm slide, two
drops of sodium chloride were added, and a cover
slide was placed while the slide was examined
under x40 magnification of the light microscope.
The motility estimate was done by taking estimates
from four different apexes of the angle and finding
the average according to Umesiobi and Iloeje,
(1999).
Sperm Concentration
A haemocytometer was used to determine the
sperm concentration. A red cell pipette was used to
suck up semen to 0.5ml mark. The semen was
diluted by sucking normal saline up to 1.01ml mark.
The pipette was gently rocked to ensure uniform
mixture. The first few drops were blown out. The
diluted semen was placed on a haemocytometer
slide and covered with cover slip. The slide was
then placed under the light microscope and viewed
under x40 magnification. Five squares were
counted and the average taken to get the semen
concentration. This is in line with the method of
Iheukwumere and Okere (1990).
Sperm Morphology
The morphology of the spermatozoa were
evaluated using Eosin-Nigrosin stain. Thin smears
of each collected semen sample were made on
slides with frosted ends on which the animal’s
details and date of collection were inscribed in
pencil. Smears were done by diluting a drop of the
individual’s semen sample with 2 – 3 drops of warm
Eosin-Nigrosin stain (pH 8.4) and drying the slide
by waving it in the air. All smears were left on a
warm stage to dry out completely. Slides were
thereafter viewed under the light microscope using
x200 magnification to find suitable area of good
quality on the smear to evaluate. Two hundred
sperm cells were counted with a counter and
evaluated as they came into view. Data were
recorded on a data capture sheet using the
classification of Rodriguez-Martinez and Barth
(2007) as adopted by the standard operating
procedure of the section of reproduction, Faculty of
Veterinary Science, Onderstepoort, University of
Pretoria. Normal sperm cells are those whose
acrosome are intact along with the neck, middle
piece and end piece. Whereas abnormal
spermatozoa are those whose acrosome, neck,
middle piece and end piece have been altered due
to injury or sperm ageing. These were observed
under the microscope during semen evaluation.
Live Sperm Percentage
A drop of semen was put on a clean warm slide; a
drop of eosin nigrosin stain was added. The two
drops were mixed using another clean slide and
observed under the microscope. The percentage
of live spermatozoa was assessed by identifying
those with intact cell membrane, from dye
exclusion or by hypotonic swelling (RodriguezMartinez and Barth, 2007).
Hormonal Assay
Blood samples (5ml each) were obtained with
needle and syringe by jugular vein puncture of the
24 rams on day 7 after the Pergonal injection, for
102
Int. Res. J. Agric. Food Sci.
testosterone, FSH and LH evaluation. Samples
were taken on the 7th day to allow the residual
effect of the drug to be observed. They were cooled
immediately in iced water and transferred to the
laboratory, refrigerated at 40C for 1 hour and the
serum separated by centrifugation at 5,000rpm for
10 minutes. The serum was stored immediately at
-200C
until
enzyme-immuno-assayed
for
testosterone, FSH and LH. The concentration of
these hormones in peripheral serum was
determined by enzyme-immune-assay (EIA)
system as described by Micallef et al. (1995) in
humans
Biochemical Constituents of Seminal Plasma
Semen samples used for estimation of
biochemical constituents of seminal plasma were
centrifuged at 15,000 rpm for 15 minutes.
Seminal plasma samples were immediately
subjected to laboratory analysis for the following
biochemical parameters; urea, non-protein
nitrogen,
glycerophosphocholine,
fructose,
ascorbic acid, sodium, potassium and
bicarbonate.
Sodium
and
potassium
concentrations were estimated with a flame
photometer on samples suitably diluted with
deionized water, while bicarbonate and urea
concentrations were determined according to the
method of Baker and Silverton (1986). Fructose
concentration in the plasma was determined
according to the procedure of Singgh (2004).
Ammonia concentration was determined
according to the procedure of Cheesbrough
(2004). Glycerophosphocholine, lactic acid and
ascorbic acid concentrations were determined
according to the method of Robert et al. (2000).
Data Analysis
Data obtained on semen characteristics,
hormonal
assay
and
seminal
plasma
constituents of WAD rams were subjected to
analysis of variance (ANOVA) using the
technique of Steel and Torrie (1980). Significant
treatment means were separated using
Duncan’s New Multiple Range Test as described
by Obi (1990).
RESULTS AND DISCUSSION
The results of gonadotrophin administration on
semen characteristics of West African Dwarf
rams are presented in Table 1. Rams on T3
recorded the highest semen volume of 1.10ml
and this differed significantly (P<0.05) from rams
on T1 (0.50) and T2 (0.50) which were similar
(P>0.05) to each other in semen volume. There
was no significant difference (P>0.05) between
rams on T3 (1.10) and those on T4 (0.80) in
semen volume. Rams on T4 were also similar
(P>0.05) to rams on T1 and T2 in semen volume.
The lowest semen volumes were observed in
rams on T1 (0.05ml) and T2 (0.50ml). The highest
semen volume in this study (1.10ml) was less
than the mean ejaculate volume (1.25 +0.7ml)
reported by Iheukwumere and Okere (1990) in
Yankasa rams of similar ages. Iheukwumere et
al. (2008) noted that methods of semen
collection, season of the year, breed, age, body
weight of animals, scrotal circumference and
frequency of semen harvest can affect ejaculate
volume in rams.
TABLE 1. Effect of Gonadotrophin on Semen Characteristics of WAD Rams.
Treatment (Pergonal, ml)
T1
T2
T3
Parameters
0.00
0.11
0.22
Semen volume (ml)
0.50b
0.50b
1.10a
Mass motility
3.00a
3.00a
3.30a
a
b
Individual motility (%)
85.00
60.00
90.00a
b
a
pH
8.00
10.00
8.00b
Sperm concentration (x10 6/ml)
0.32b
0.50a
0.79a
a
a
Proportion of live sperm cells (%)
74.52
78.70
80.20a
a
a
Proportion of normal sperm cells (%)
72.32
75.25
77.30a
b
b
Proportion of dead sperm cells (%)
25.48
21.30
19.80b
ab Means
SEM:
within row having different superscript are significantly different
Standard error of mean
(P<0.05)
T4
0.33
0.80ab
2.00b
70.00ab
10.00a
0.61a
60.10b
50.25b
39.90a
SEM
0.14
0.31
6.88
0.58
0.10
4.59
6.25
4.41
Egu and Ukpabi 103
Rams on T3 recorded the highest value for mass
motility (3.30). The mean value for mass motility
in rams on T1 was similar (P> 0.05) to the values
for mass motility in rams on T2 and T3 except T4
and this differed significantly (P<0.05) from rams
on T4 (2.00). However, there were no significant
differences (P>0.05) among rams on T3 (3.30),
T2 (3.00) and T1 (3.00) which were similar to each
other in mass motility. Mass motility ranges from
1 to 5 (Brackett, 2005).
Rams on T3 had significantly increased individual
motility than rams on T1, T2 and T4. Rams on T2
(60.00%) were similar (P>0.05) to rams on T 4
(70.00%). However, there were no significant
differences (P>0.05) among rams on T1
(85.00%), T3 and T4. The highest score in
individual motility obtained in this study (90.00%)
was higher than 85.0 ± 7.5% reported by
Iheukwumere et al. (2001) in Yankasa rams of
similar ages. This could be attributed to high
capacity for induction of spermatogenesis and
improvement of fertility by Pergonal injection.
Sperm motility is a critical indicator of semen
quality and fertility potential because it is required
for penetration of cervical mucus, transport
through the female genital tract, and penetration
through the Corona radiata and Zona pellucida
before oocyte fertilization (Iheukwumere, 2005).
Sperm motility is also affected by frequency of
semen collection (Iheukwumere et al., 2008).
The group of rams on T2 and T4 recorded the
highest values in pH (10.00) each and this
differed significantly (P<0.05) from rams on T 1
and T3 which were similar (P<0.05) to each other
in semen pH (8.00). The highest semen pH
values obtained in this study were higher than
the normal range (7-8) as also reported by
Meacham (2002) in humans. The measured pH
can depend on the length of time since
ejaculation and it tends to increase shortly after
ejaculation as a result of loss of CO2 (Meacham,
2002). Semen maintains its pH near neutral in
the acidic vaginal environment, providing the
sperm with the opportunity to enter the neutral
pH of cervical mucus (Meacham, 2002).
Rams on T3 recorded the highest value for sperm
concentration (0.79 x106/ml) and this differed
significantly (P<0.05) from rams on T1 (0.32 x
106/ml). However, there were no significant
differences (P>0.05) among rams on T2 (0.50
x106/ml), T3 (0.79x106/ml) and T4 (0.61 x106/ml)
which were similar (P>0.05) to each other in
sperm concentration. The highest sperm
concentration obtained in this study (0.79
x106/ml) was similar to the highest sperm
concentration (0.77±0.07x106/ml) reported by
Iheukwumere et al. (2001) in Yankasa rams of
similar ages. This was within the normal range of
200 to more than 1,000 million spermatozoa/ml
reported by Rodriguez-Martinez and Barth
(2007). Normally an increase in the semen
collection frequency is associated with a
decrease
in
spermatozoa
concentration
(Iheukwumere and Okere, 1990; Arroita et al.,
2000).
There were no significant differences (P>0.05)
among rams on T1 (74.52%), T2 (78.70%) and T3
(80.20%) which were similar (P>0.05) to each
other in percentage of live sperm cells but rams
on T3 differed significantly (P<0.05) from rams on
T4 (60.10%). The highest percentage of live
sperm cells obtained in this study was within the
normal range (77.2 ± 9.7 to 79.4 ± 4.9%) as also
reported by Iheukwumere et al. (2001) in Nigeria
Yankasa rams. Percentage of normal sperm
cells followed the same pattern as live sperm
cells. Rams on T3 recorded the highest
percentage of normal sperm cells (77.30%) and
this differed significantly (P<0.05) from rams on
T4 (50.25%). However, there were no significant
differences (P>0.05) among rams on T1
(72.32%), T2 (75.25%) and T3 (77.30%) which
were similar (P>0.05) to each other in
percentage of normal sperm cells. The highest
percentage in normal sperm cells obtained in the
study (77.30%) was lower than the range (82.4 ±
3.8 - 87.0 ± 8.2%) as reported by Iheukwumere
et al. (2001) in Nigeria Yankasa rams. This could
be attributed to differences in breed. The West
African dwarf ram is the smallest of the Nigerian
breeds (Oni, 2002).
Rams on T4 recorded the highest percentage of
dead sperm cells (39.90%) and this differed
significantly (P<0.05) from rams on T1 (25.48%),
T2 (21.30%) and T3 (19.80%) which were similar
(P>0.05) to each other in percentage of dead
sperm cells. The lowest value in dead sperm
104
Int. Res. J. Agric. Food Sci.
cells was recorded by rams on 0.22ml
gonadotrophin (T3).
The observation that the group that received the
highest dose of Pergonal recorded the lowest
percentage of live sperm cells, normal sperm
cells and highest percentage of dead sperm cells
suggests that 0.33ml gonadotrophin/ram (T 4)
administered within 3 days in this study could
have deleterious effect on sperm cells.
The results of gonadotrophin administration on
hormonal profile of West African Dwarf rams are
shown in Table 2.
TABLE 2: Effect of Gonadotrophin on Hormonal Profile of WAD Rams.
Treatment (Pergonal, ml)
T1
T2
T3
Parameters
0.00
0.11
0.22
Follicle stimulating hormone (ng/ml)
10.10b
19.30a
10.35b
Luteinizing hormone (ng/ml)
4.20
4.18
4.18
Testosterone (ng/ml)
0.95a
0.75a
0.65b
T4
0.33
10.30b
4.18
0.60b
SEM
2.26
0.01
0.08
ab Means
in the same row with different superscripts are significantly (P<0.05) different.
SEM = Standard error of mean.
Rams on T2 recorded the highest FSH
concentration (19.30 ng/ml) and this differed
significantly (P<0.05) from rams on T3 (10.35
ng/ml), T4 (10.30ng/ml) and T1 (10.10 ng/ml)
which were similar (P>0.05) to each other in FSH
values. The observation in this study that the
FSH value in the rams treated with 0.11ml
gonadotrophin (T2) was higher (P<0.05) than in
the other groups that received higher doses of
the drug suggest that a high dose of the drug
such as 0.22ml or 0.33ml gonadotrophin/ram
within 3 days as in this study could excite
suppressive effects on the hypothalamus. This
observation is in agreement with the report of
Iheukwumere (2005) in goats.
There were no significant differences (P>0.05)
among the treatment groups in luteinizing
hormone (LH) levels in the serum. Rams on the
control treatment recorded the highest value in
LH (4.20 ng/ml). Similarity (P>0.05) in LH values
among the treatment groups were an indication
that the gonadotrophin injections had no
deleterious effects on LH secretion. It is common
knowledge that LH as an interstitial cell
stimulating hormone (ICSH) stimulates the
interstitial cells of leydig to produce testosterone
which facilitates the process of spermatogenesis
(Herbert et al., 2000).
Rams on T1 recorded the highest testosterone
value (0.95ng/ml) and this differed significantly
(P<0.05) from the values in rams on T3
(0.65ng/ml) and T4 (0.60ng/ml) which were
similar (P>0.05). However, there was no
significant difference (P>0.05) between rams on
T1 and T2 (0.75ng/ml) in testosterone
concentrations. The lowest testosterone value of
(0.60ng/ml) was observed in ram on 0.33ml
Pergonal (T4). Testosterone values obtained in
this study were within the range of 0 – 20ng/ml
reported in sheep by King et al. (1993). In a
similar study, Herbert et al. (2000) had reported
differences in the serum testosterone levels that
showed slightly higher values for the Clomid®
treated groups than the control group but were
not significantly different (P>0.05). The
observation in this study that the testosterone
value in the control group (T1) was higher
(P<0.05) than in the groups that received
Pergonal injections suggest that the doses of
gonadotrophin administered in this study (0.11,
0.22 and 0.33ml gonadotrophin / ram) within 3
days could excite suppressive effects on the
hypothalamus that caused progressive decrease
in the testosterone levels. It has also been
reported that exogenous administration of
testosterone itself leads to an inhibitory effect on
the hypothalamus thus reducing the sperm
production process (Adamopoulous et al., 1990;
Li-Jian et al., 1997; Mungai et al., 1997).
The results of gonadotrophin administration on
biochemical constituents of seminal plasma of
WAD rams are presented in Table 3.
Egu and Ukpabi 105
TABLE 3: Effect of Gonadotrophin on Biochemical Constituents of the Seminal Plasma of WAD rams
Treatment (Pergonal, ml)
T1
T2
T3
T4
Parameters
0.00
0.11
0.22
0.33
SEM
Urea (mg/100ml)
30.00b
36.10b
40.00ab
42.00a
1.50
Ammonia (mg/100ml)
2.00
2.00
2.00
2.00
0.00
Lactic acid (mg/100ml)
40.00
40.00
40.00
40.00
0.00
Glycerophosphocholine (mg/100ml)
205.00b
220.00b 240.00ab 250.00a
4.69
b
b
a
a
Fructose (mmol/L)
480.00
480.00
500.00
500.00
5.77
Ascorbic acid (mg/100ml)
5.20b
5.60b
6.00b
8.00a
0.52
Sodium (mmol/L)
40.54b
43.15a
38.10b
40.28b
1.03
Potassium (mmol/L)
3.48b
5.45a
5.34a
5.37a
0.4
a
b
b
b
Bicarbonate (mmol/L)
21.30
20.65
20.70
20.83
0.15
abc
Means within row having different superscripts are significantly different (P<0.05)
SEM = Standard error of mean
The highest value for urea (42.00mg/100ml) was
recorded in rams on T4 and this differed
significantly (P<0.05) from rams on T1
(36.00mg/100ml) and T2 (36.00mg/100ml) which
were similar (P>0.05) to each other and also
similar (P>0.05) to rams on T3 (40.00mg/100ml) in
urea value. However, there was no significant
difference (P>0.05) between rams on T4 and T3 in
urea values. Cortada et al. (2000) reported that a
sharp increase in plasma urea level could result in
gonadal degeneration and infertility with reduced
sperm production and loss of libido.
There were no significant differences (P>0.05)
among the treatment groups in ammonia and
lactic acid which recorded 2.00 (mg/100ml) and
40.00 (mg/100ml) respectively across the
treatments. Owen and Katz (2005) inferred that
an interaction between lactic acid and CO2
concentration can lead to pH changes.
Rams on T4 recorded the highest value
(250.00mg/100ml) in glycerophosphocholine and
this differed significantly (P<0.05) from rams on T1
(205.00mg/100ml) and T2 (220mg/100ml) which
were similar (P>0.05) to each other and also
similar (P>0.05) to rams on T3 (240.00mg/100ml).
However, there was no significant difference
(P>0.05) between rams on T4 and T3 in
glycerophosphocholine values. The level of
glycerophosphocholine in all the groups indicates
that sperm maturation was enhanced by the
administration of gonadotrophin injection (Kidd,
2005). Rams on T3 and T4 recorded the highest
fructose concentration in the seminal plasma
(500.00mg/100ml) and this differed significantly
(P>0.05) from rams on T1 and T2 which were
similar (P>0.05) to each other in fructose level
(48.00mg/100ml). This implies that gonadotrophin
may only affect fructose levels at higher doses.
Gonzales et al. (1993) reported a wide range of
136 – 628mg/100ml fructose for semen. Owen
and Katz (2005) reported that fructose is a
measure of seminal vesicle function, being a
source of energy for the sperm. Fructose is the
primary source of lactic acid in semen.
Ascorbic acid concentration increased with
increased levels of gonadotrophin administration.
The values ranged from 5.20 to 8.00 (mg/100ml).
The value for rams on T4 differed significantly
(P<0.05) from the values for rams on T3, T2 and
T1 which were similar (P>0.05) to each other.
Studies have shown that vitamin C plays a vital
role in increasing semen volume, sperm
concentration and motility in rams and goats
(Sonmez and Demirci, 2003; Fazeli et al., 2010)
and keeping them strong by protecting them from
free radicals (Dawson et al., 1992; Fazeli et al.,
2010). Results of this study indicate that the
administration of gonadotrophin in rams
enhanced the concentration of ascorbic acid in the
seminal plasma which is very vital in assessing
semen quality and fertility in male animals.
There were significant differences (P>0.05)
among the treatment groups in sodium,
potassium and bicarbonate levels in the seminal
plasma. Sodium and potassium were both highest
when T2 was administered. The highest values
were 43.15 (mmol/l) recorded for sodium and
5.45mmol/l or potassium. The lowest value for
106
Int. Res. J. Agric. Food Sci.
sodium was 40.28mmol/l recorded by rams on T4.
A positive and significant correlation has been
established between sodium concentration and
sperm concentration in rams (Akpa et al., 2013).
For potassium, the lowest value was 3.48mmol/l
recorded for rams on T1 indicating that the
administration of gonadotrophin increased this
element. Sperm concentration is also positively
correlated with potassium concentration in
seminal plasma. This trend was equally observed
in this study. Increasing potassium concentration
in the seminal plasma is negatively correlated with
progressive motility of sperm in rams and bucks
while sodium has the opposite effect (AbdelRahman et al., 2000; Akpa et al., 2013).
Rams on T1 recorded the highest bicarbonate
value (21.30mmol/l) and this differed significantly
(P<0.05) from rams on gonadotrophin treatments
which were similar (P>0.05) to each other. The
lowest value in seminal bicarbonate was
observed in rams on T2 (20.65mmol/l). The values
obtained in gonadotrophin treated groups ranged
from 20.65 to 20.83(mmol/l) and were similar to
the value (20.00mmol/l) recorded by Okamura et
al. (2006), who also inferred that sodium
bicarbonate in seminal plasma stimulates sperm
motility.
CONCLUSION
The results of this study indicate that the
administration of up to 0.33ml of gonadotrophin
enhanced sperm quality of WAD rams. The main
intention of the administration of gonadotrophin
was to stimulate spermatogenesis and improve
semen quality, a higher dosage tends to have a
suppressive effect on the number of live and
normal sperm cells and the concentration of
testosterone in the seminal plasma of WAD rams.
Though most of the values obtained fall within the
normal ranges for adult sheep, there is need to
constantly monitor blood and hormonal profiles of
West African dwarf rams under gonadotrophin
treatment for spermatogenesis.
CONFLICT OF INTEREST
The authors have declared no conflict of interest.
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