INFLUENCE OF SEMEN QUALITY ON SPERM
SURVIVABILITY, SPERM DNA FRAGMENTATION
AND FERTILITY RATE OF THE MALE
DROMEDARY CAMELS
Zeidan, A.E.B.; E.A.A. Ahmadi ; A.M. Amer ;
T.M.M. Mahdy and A.L.I. Desoky.
Animal Production Research Institute, Dokki, Giza, Egypt.
Key Words: Dromedary camels, Semen quality, Blood, DNA
fragmentation, Conception rate.
ABSTRACT
Two experiments were carried out. Twelve Maghrebi camels at <6 to
10 years of age were used in the present study. Semen samples were
collected using an artificial vagina (AV) and divided into their qualities to
three groups (Poor ≥20-40% motile spermatozoa, Normal, ≥40-60% motile
spermatozoa and Good, ≥60-80% motile spermatozoa). Copulation time,
semen characteristics, sperm DNA fragmentation Index, sperm mensuration
and blood constituents of the male dromedary camels were recorded
(Experiment 1). Different semen qualities were diluted with Lactose-Yolk-
Citrate (LYC) extender and stored at 5°C for 3 days (Experiment 2).
Conception rates (%) of she-camels artificially inseminated with different
semen qualities were assessed.
The obtained results showed that, copulation time (min) and semenejaculate
volume (ml) were significantly (P<0.05) better in camels
containing Normal and Good quality than Poor quality of semen. Semen
colour was Thin creamy, Creamy and Creamy whereas semen consistency
was Semi-viscous, Viscous and Viscous for Poor, Normal and Good quality
of the dromedary camel semen, respectively. Furthermore, seminal pH value
and sperm mensuration (μm) of the dromedary camels were insignificantly
differences in different semen qualities. The percentage of sperm motility
and sperm-cell concentration (x106/ml) were significantly (P<0.05)
increased, while the percentages of dead spermatozoa, abnormal
spermatozoa, acrosome damage, sperm chromatin damage and sperm DNA
fragmentation Index were significantly (P<0.05) decreased of the camels
with Normal and Good quality than Poor quality of the dromedary camel
semen. Moreover, total protein, albumin and globulin (gm/100ml)
concentrations showed insignificantly higher, while total cholesterol
concentration (mg/100ml) was significantly (P<0.05) higher in the camels
containing Normal and Good quality than Poor quality of semen. On the
other hand, Sodium concentration (mg/100ml) and activity of aspartateaminotransaminase
(AST) and alanine-aminotransaminase (ALT) enzymes
increased significantly (P<0.05) in the camels having Poor quality compared
Egypt. J. of Appl. Sci., 35 (5) 2020 28-51
to those with Normal and Good quality of semen. Similarly, Potassium
(mg/100ml), Calcium (mg/100), Total phosphorus (mg/100ml), Zinc
(μg/100ml) and testosterone concentrations (ng/100ml) were significantly
(P<0.05) higher in the camels having Normal and Good quality than Poor
quality of semen (Experiment 1). The percentages of sperm motility and
sperm storagability of the diluted spermatozoa with LYC extender were
significantly (P<0.05) higher in the camels with Normal and Good quality of
semen, while the percentages of dead spermatozoa, abnormal spermatozoa,
acrosome damage and sperm chromatin damage and activity of AST and
ALT enzymes were significantly (P<0.05) lower in the camels having
Normal and Good quality than Poor quality of the camel semen stored at
5°C (Experiment 2). The advancement of storage times was decreased
significantly (P<0.05) semen quality in different qualities of semen with the
successive storage times at 5°C for 3 days. Conception rates (%) of shecamels
artificially inseminated with different qualities were significantly
(P<0.05) lower with Poor quality than Normal and Good quality of semen.
In conclusion, copulation time, semen characteristics, sperm mensuration,
sperm DNA fragmentation Index, blood constituents, sperm survivability
and fertility rates showed better for Normal and Good quality than Poor
quality semen.
INTRODUCTION
The camel (Camelus dromedaius) is an important livestock species
that can uniquely adapted to hot and arid environments. There is an urgent
need to develop food production resources in Africa and other developing
countries to solve the dramatic increasing in human population and
insufficient amount of meat and milk.
Maximizing the use of outstanding sires in artificial breeding depends
on many factors. Developing and extender that preserves the high fertility of
unfrozen semen for a long time would help increase the overall conception
rate, encourage more complete use of semen and make semen from
outstanding sires more widely available in management systems using fresh
semen (Foote and Bratton, 1960).
The blood constituents are the mirror which reflects the healthy
condition of animals. So, the biochemical studies under different fluctuating
climatic conditions are very important for clinicians in the field during
interpretation of their findings. Minerals and trace elements has long been
known to be important in animal nutrition as they may be dietary essential
and vital to enzyme processes of living cells or have some metabolic
activity, bone formation and reproductive performance (Zeidan et al., 2001
and Matter, 2019).
Media known to prolong the survival of fresh bovine spermatozoa
contain complex buffers (Foote and Bratton 1960), and varying levels of
egg yolk, glucose, glycine, glycerol, citrate and other salts (Foote and
29 Egypt. J. of Appl. Sci., 35 (5) 2020
Bratton 1960). It has long been recognized that the electrolyte composition,
buffering agent and osmotic pressure of the solution.
Achievement of the high reproductive activity partially depends on the
success of artificial insemination (AI) which in turn is dependent on semen
quality obtained and its capacity for dilution and storage with minimum loss
of fertilizing ability (Wilson, 1984). Generally, the live spermatozoa can be
prolonged for several days at chilled storage (2-5°C). However, satisfactory
fertility results are not always achieved after as little as one day of storage
(Murase et al.,1990 and Zeidan et al., 2001).
Semen characteristics were considered inconstant predictors of
reproductive efficiency especially with very poor quality (Rodriguez-
Martinez, 2003). Artificial insemination programs is focusing on the
development of accurate methods that have been able to predict field fertility
with frozen-thawed semen. Indeed, more sensitive methods are needed to
enable elimination of the low fertility in the high selected bull (Hallap et al.,
2005). Semen quality must consider not only sperm motility and viability
but also sperm functional competence like capacitation, sperm chromatin
damage and sperm DNA fragmentation Index. Studies in humans indicate
that the frequency of spermatozoa containing fragmented DNA may be a
new independent parameter of semen quality and fertility (Evenson et al.,
2002). A new procedure for the sperm DNA fragmentation Index has been
developed in human sperm (Frenandez et al., 2003) and in mouse
(Rodriguez et al., 2005) based on the sperm chromatin damage test (Enciso
et al., 2006).
The objective of the present work was to study the effects of
copulation time, semen characteristics, sperm mensuration, blood
constituents, sperm chromatin damage and sperm DNA fragmentation
Index in the different semen qualities of dromedary camels (Experiment
1). Evaluation of different semen qualities and sperm survivability of the
diluted camel semen with LYC extender during storage at 5°C for 3 days
in different qualities of semen was recorded (Experiment 2). Conception
rates (%) of she-camels artificially inseminated with different semen
qualities were also assessed.
MATERIALS AND METHODS
The experimental work was carried out in the Laboratory of Public
Health, Faculty of Veterinary Medicine, Zagazig University, Zagazig, and
Private Camel Farm, Marsa Motrouh Governorate, Egypt, during the period
from December, 2019 to October, 2020.
Two experiments were carried out. The objective of the first
experiment was to study the effects of copulation time, semen
characteristics, sperm mensuration, blood constituents, sperm chromatin
damage and sperm DNA fragmentation Index in different semen qualities in
the dromedary camels. After collection, semen was evaluated and the
Egypt. J. of Appl. Sci., 35 (5) 2020 30
ejaculates were diluted with LYC extender could be considered as the Poor
quality (consistently ≥20-40% motile spermatozoa, Normal ≥40-60% motile
spermatozoa and Good ≥60-80% motile spermatozoa). The second
experiment aimed to define the effects of semen quality on survivability and
activity of AST and ALT enzymes of the camel spermatozoa diluted with
LYC extender during storage at 5°C for 3 days. Conception rates (%) of shecamels
artificially inseminated with different semen qualities were recorded.
1. Materials:
1.1. Experimental animals :
Twelve Maghrebi camels (Camelus dromedarius) aging < 6-10 years
old and 500-600 kg live body weight, were used in the present study. All
camels were in healthy condition.
1.2. Feeding and management:
The rations offered to camels were calculated according to Banerjee
(1988). Clean fresh water was offered freely to all camels.
2. Methods:
2.1. Camel semen collection:
Eight ejaculates were collected and evaluated from each camel during
the breeding season according to Abd El-Raouf et al. (1975) at 0.8:00 and
10.00 a.m. using an artificial vagina (AV) as the method described by
Zeidan (2002). The AV was filled with water at 50-55°C and the
temperature inside the inner liner was stabilized at 45-50°C. The ejaculates
were usually comes in fractions. Fresh camel semen that has a jelly-like
consistency is left for liquefaction for about 30-60 minutes to make the
sperm attained motility.
2.2. Semen extension :
Semen samples were collected, pooled and evaluated for each camel
and then diluted with lactose-yolk-citrate (LYC) extender (2.9g sodium
citrate dehydrate, 0.04g citric acid anhydrous, 1.25g lactose and 10ml eggyolk
per 100ml distilled water, 500 I.U/ml penicillin and 500 μg
Streptomycin sulphate) according to Musa et al. (1992). Semen extension
was carried out by adding the appropriate volume of the semen slowly to the
extender as the method described by Salisbury et al. (1978). The dilution
rate was 1 ml semen : 3 ml extender accoding to Musa et al. (1992).
Semen samples were immediately diluted with LYC extender and kep
at 25-30ºC for liquefaction in waterbath for 45 mins, where semen samples
were shaked throughly at this time. Thereafter, the mixture was transported
in glass containers to a cooled chamber cabinet at 5ºC for 3 days.
2.3. Chilling of semen at 5ºC :
The test tubes containing extended semen for each camel were placed
in a 500ml beaker containing water at 30ºC with a thermometer in order to
facilitate periodic check of the temperature during cooling period. Another
test tubes containing extended semen only were placed in the beaker to
31 Egypt. J. of Appl. Sci., 35 (5) 2020
maintain the extended temperature similar to that of semen (all the test tubes
were covered with dark plastic sheath). The beaker was placed in a
refrigerator and gradually cooled till their temperature reached to 5ºC during
a period of 1.5-2.0 hours according to Musa et al. (1992). The cooled
spermatozoa were kept at 5ºC for up to 3 days. After each storage time
(0,1,2 and 3 days), percentages of sperm motility, dead spermatozoa,
abnormal spermatozoa, acrosome damage and chromatin damage of
spermatozoa were recorded.
1. Copulation time (minutes):
Duration of copulation was measured from the time of penile
intromission into the artificial vagina until withdrawal as the method
described by Bravo et al. (2000).
2. Semen characteristics:
2.1. Semen colour:
Semen colour was determined by direct visual examination from the
collecting tube (Bravo et al., 2000).
2.2. Semen consistency:
Semen consistency was qualified as viscous when semen did not drop
from a Pasteur pipette, semi-viscous when some semen dropped from the
Pasteur pipette to glass slide and liquid when semen was fluid and dropped
readily from the Pasteur pipette according to Bravo et al. (2000).
2.3. Semen-ejaculate volume (ml):
Semen-ejaculate volume was determined using a conical graduated tube.
2.4. Hydrogen-ion concentration (pH):
Seminal pH value in the male dromedary camels was measured using
universal indictor paper and standard commercial stains according to
Karras (1952).
2.5. Sperm motility (%):
Generally, camel sperm motility (%) was detected as an oscillatory
motion the flagellum but not progressive due to the viscous materials
according to Campbell et al. (1956). With regard to extended semen,
percentage of sperm motility was determined using one drop of the extended
semen after each storage period. The drop of the extended semen was
covered by a warmed cover slip and immediately examined using high
power magnification (400x).
2.6. Storagability (%):
Storagability (%) of the cooled camel spermatozoa refers to the
percentage of original motile spermatozoa still motile after 3 days of storage
time at 5oC as the method described by Yassen and El-Kamash (1970).
2.7. Dead spermatozoa (%) :
The eosin/nigrosin staining procedure was carried out by dissolving
1.67gm eosin and 10.00 gm nigrosin in distilled water up to 100 ml
according to Hackett and Macpherson (1965).
Egypt. J. of Appl. Sci., 35 (5) 2020 32
2.8. Abnormal spermatozoa (%) :
The percentage of abnormal spermatozoa (%) was determined in the
same smears prepared for live/dead spermatozoa ratio (Watson, 1975).
2.9. Acrosome damage of spermatozoa (%) :
The percentages of acrosome damage (%) of spermatozoa was
estimated according to Watson (1975).
2.10. Sperm-cell concentration (x106/ml):
Sperm-cell concentration (x106/ml) was counted using
haemocytometer according to Khan (1971).
2.11. Chromatin damage of spermatozoa (%):
Toluidine blue staining was performed as the method described by
Erenpreiss et al. (2004). Smears were fixed in ethanol-acetic acid glactial
(3:1, v/v) for 1 min and 70% ethanol for 3 mins. Smears were hydrolyzed
for 20 mins in 1 Mm HCL, rinsed by distilled water and air-dried. One
droplet of 0.025% Toluidine blue in Mcllvaine buffer. Smears were
evaluated with light microscope magnification (x1000). Percentage of
chromatin damage was estimated by evaluating 300 spermatozoa in each
smear. Spermatozoa stained as green to light blue were considered to have
normal chromatin, while those stained dark blue to violet were considered to
have damaged chromatin.
2.12. Sperm DNA fragmentation Index (%):
Single and double-stranded sperm DNA fragmentation Index can be
measured using the alkaline and neutral comet assays. The procedure was
simultaneously performed in semen samples on two distinct slides.
Different semen qualities were washed with HEPES/MOPS buffer. Then,
25-μl aliquotes of spermatozoa was arranged on two pretreated slides of
agarose gel and placed on a cold plate at 5°C for 4 minutes. Thereafter,
the cover slips were carefully isolated, and the slides were washed with an
excess of lysing buffer for 30 minutes, followed by a 10 minute wash in
Tris-borate EDTA (TBE). In the neutral comet assay, electrophoresis was
performed with a TBE arrangement of 20 V (1 V/cm) for 12 minutes and
30 seconds, with a subsequent wash with 0.9% NaCl for 2 minutes. The
slide was washed for 3 minutes at 5°C and electrophoresis was then
performed in 0.03M NaOH at 20V (1 V/cm) for 4 minutes. Then, both
slides were incubated in a neutralizing solution for 5 minutes with TBE
for 2 minutes. The slides were dried in a graded series of ethyl alcohol
solutions (75%, 85%, and 100%) for 2 minutes each. Finally, 300 Poor,
Normal and Good quality of spermatozoa were assessed according to
Enciso et al. (2011).
3. Sperm mensuration (μm):
Mensuration of spermatozoa was measured using calibrated eye-piece
micrometer scale using Eosin-Nigrosin stain according to Campbell et al.
(1956). Every pixel of the micrometer scale was represented of 0.085 μm
33 Egypt. J. of Appl. Sci., 35 (5) 2020
when an oil immersion lens (x100). The parameters of the mensuration of
spermatozoa included length and width of sperm head as well as length and
width of sperm tail according to Banaszewska et al. (2011).
4. Blood serum constituents:
Blood samples were collected pre-slaughter from jugular vein in the
non-heparinized vacutainer tube for each camel in different ages and
centrifuged for 15 minutes at 8000 RCF. Serum samples were taken weekly
and stored at -20°C until analysis. Total protein, Albumin, Globulin, Total
cholesterol, Sodium, Potassium, Calcium, Total phosphorus, Zinc and
Testosterone concentrations and activity of AST and ALT enzymes were
recorded. Total protein, albumin and total cholesterol concentrations were
determined colourimetrically according to the method described by Tietz
(1982). Globulin level was calculated by subtraction of albumin content
from the total protein content. Testosterone hormone concentration (T2) was
determined by Radioimmunoassay Technique (RIA) of Coa-Ab-Cont Kits
(Diagnostic Products Corporation-Los Anglos, USA) according to
Abraham (1977). Sodium, Potessum, Calcium, and Total Phosphorus
concentrations were determined colourimetrically according to Tietz (1982).
Zinc concentration was determined using 5P9 Atomic Absorption
Spectrophotometry (Pye Unicam) as the method described by Willis (1960).
5. Biochemical analysis in seminal plasma (U/106 spermatozoa):
After each storage times (0, 1, 2 and 3 days), the cooled semen was
centrifuged for 15 minutes at 8000 RCF. Seminal plasma was separated and
stored at-20°C until assay of enzymes. Activity of aspartate
aminotransaminase (AST) and alanine-aminotransaminase (ALT) enzymes
(Spectroliv-UV Auto, LIU-2602, Labomed, USA) were recorded as the
method described by Reitman and Frankle (1957).
6. Fertility rate (%):
Semen was deposited into the uterus using an insemination gun to the
internal cervical. Insemination has to be done using a porcine rubber
insemination tube heated to 38°C to prevent cold shock.
She-camels were injected intramuscularly with 3000 IU of human
chorionic gonadotropin (hCG) in the from of Proface (1ml-ampules packed
by EPICO, Egypt, under licence from I.F. Serna, Rome, Italy) to induce
ovulation (Anouassi et al., 1994). She-camels were artificially inseminated
approximately 48 hrs post hCG injection.
She-camels were divided into three groups as follows:
Group 1 (n-17) : She-camels were artificially inseminated with Poor quality
semen (2ml) containing 100x106 motile spermatozoa.
Group 2 (n-18) : She-camels were artificially inseminated with Normal
quality semen (2ml) containing 100x106 motile
spermatozoa.
Egypt. J. of Appl. Sci., 35 (5) 2020 34
Group 3 (n-18) : She-camels were artificially inseminated with Good quality
semen (2ml) containing 100x106 motile spermatozoa.
Pregnancy rate was diagnosed at 60 days after the date of insemination.
3. Statistical analysis :
Data were statistically analyzed by one-way and two-way design
(ANOVA) using General Linear model (GLM) procedure of SAS (SAS,
2006). Duncan's multiple range test (Duncan, 1955) was used to detect
significant differences among means. Percentage values were transformed to
arc-sin values before being statistically analyzed. Conception rates were
analyzed by Chi-square test.
The following model used was as follows:
The first experiment :
Yij = μ + qi + ej
Yij = is the observed value of the dependent variable determined from a
sample
taken from each animal.
μ = is the overall mean.
qi = is the fixed effect of semen quality.
ej = is the residual error
The second experiment :
Yijk = μ + qi + Sj + (qi x Sj) + eijk
Yijk = is the observed value of the dependent variable determined from a
sample
taken from each animal.
μ = is the overall mean.
qi = is the fixed effect of semen quality.
Sj = is the fixed effect of storage time
(qiXSj) = is the first order interaction between semen quality and
storage time.
e ijk = is the residual error.
RESULTS AND DISCUSSION
The first experiment:
1. Copulation time (minutes):
Data presented in Table 1 showed that copulation time was
significantly (P<0.05) better in the dromedary camel semen having Normal
and Good quality than Poor quality of semen. It is generally increase of
steroid hormone secretion, consequently increase of testosterone
concentration which stimulate libido or copulation time, so this reflected on
semen quality. These results are partially agreement with those of Zeidan et
al. (2001) in the male dromedary camels.
2. Semen characteristics:
2.1. Semen colour :
Semen colour was Thin creamy, Creamy and Creamy in the camels
with Poor, Normal and Good semen quality, respectively (Table 1). The
35 Egypt. J. of Appl. Sci., 35 (5) 2020
difference colour of semen quality may be due to the different
concentrations of spermatozoa and semen consistency. Similar trends were
recorded by Zeidan et al. (2001) in the male dromedary camel.
2.2. Semen consistency:
Semen consistency was Semi-viscous, Viscous and Viscous for
camels with Poor, Normal and Good quality, respectively (Table 1).
Viscosity of semen is usually attributed to the presence of
mucopolysacharides (Garnica et al., 1993) which can be secreted from
bulbourethral glands or the prostate gland.
2.3. Semen-ejaculate volume (ml):
Semen-ejaculate volume showed significantly (P<0.05) higher of the
dromedary camels having Normal and Good quality than Poor quality of
semen (Table 1). Similar trends were recorded by Zeidan et al. (2001) and
Matter (2019) in the dromedary camel.
2.4. Hydrogen-ion concentration (pH):
Data presented in Table 1 showed that seminal hydrogen-ion
concentration (pH) value was insignificantly different in the dromedary
camel with different qualities of semen (Poor, Normal and Good).
2.5. Percentage of sperm motility:
The percentage of sperm motility was significantly (P<0.05) higher of
the male dromedary camel having Good quality than Poor and Normal
quality of semen (Table 1). Similarly, the percentage of sperm motility was
significantly (P<0.05) higher in the dromedary camel semen having Good
quality than Poor and Normal quality of semen. These results may be due to
the hypoactive of Leydig cells which are considered to be testosterone
hormone producing factor, so this reflected on semen quality. Similar trends
were recorded by Zeidan et al. (2001) and Matter (2019) in the male
dromedary camels.
2.6. Percentage of dead spermatozoa :
Data presented in Table 1 showed that the percentage of dead camel
spermatozoa was significantly (P<0.05) higher of the male dromedary camel
semen in the camels with Poor quality than Normal and Good quality of
semen. The highest (P<0.05) value of the percentage of dead spermatozoa of
the Poor quality of semen may be attributed to the disturbance in
spermatogenesis process or even death of spermatozoa which reflected on a
bad semen quality. Similar trends were recorded by Zeidan et al. (2001)
and Matter (2019) in the male dromedary camels.
2.7. Percentage of abnormal spermatozoa:
The percentage of abnormal spermatozoa was significantly (P<0.05)
higher of the male dromedary camel semen having Poor quality than
Normal and Good quality of semen (Table 1). The highest (P<0.05) value of
percentage of abnormal camel spermatozoa was recorded with Poor quality
and the lowest (P<0.05) value was recorded with Good quality of semen.
Egypt. J. of Appl. Sci., 35 (5) 2020 36
2.8. Percentage of acrosome damage:
The percentage of acrosome damage was significantly (P<0.05)
higher of the dromedary camel having Good quality than Normal and Poor
quality of semen (Table 1). The highest (P<0.05) value of the percentage of
acrosome damage of spermatozoa was recorded with Poor quality of semen
and the lowest (P<0.05) value was recorded with Good quality in the
dromedary camel semen.
2.9. Percentage of chromatin damage:
Data presented in Table 1 revealed that the percentage of
chromatin damage of spermatozoa in the dromedary camels was
significantly (P<0.05) decreased with the Normal and Good quality of
camels compared to those with Poor quality of semen. These results may
be attributed to the decrease of adenosine triphosphate which activated
apparently ability of resynthesizing. This was accompanied with
precipitation full in the rate of fructolysis, consequently increased
chromatin damage (Mann and Lutwak-Mann, 1981).
2.10. Sperm-cell concentration (x106/ml):
Sperm-cell concentration was significantly (P<0.05) higher of the
dromedary camel having Normal and Good quality than Poor quality of
semen (Table 1). The highest (P<0.05) value of sperm-cell concentration
(x106/ml) was recorded with Normal and Good quality of semen, while the
lowest (P<0.05) value was recorded with Poor quality of semen in the
dromedary camels. Similar trends were recorded by Zeidan et al. (2001)
and Matter (2019) in the male dromedary camels. These results may be due
to the hyperactive of Leydig cells which are increase of testosterone
hormone secretion which more in the camels having Normal and Good
quality than Poor quality of semen.
2.11. Sperm DNA fragmentation Index (%):
The percentage of sperm DNA fragmentation Index of the dromedary
camels was significantly (P<0.05) decreased in the camels having Normal
and Good quality of semen compared to those with Poor quality of semen
(Table 1). In fact sperm abnormalities could finally produce DNA
degradation as part of an apoptotic process to eliminate defective sperm
cells. Consequently, most of the morphologically abnormal and DNA
fragmented spermatozoa present in the ejaculate.
On the other hand, the morphological abnormalities might potentially
have a genetic origin. Sperm DNA damage could be considered as the
physiological basis of some morphological defects. Some investigators
confirmed that certain aspects of fertility including morphological
abnormalities are probably under genetic control (Chenoweth, 2005).
Moreover, as morphologically normal spermatozoa can also present
with DNA damage, the inclusion of the assessment of DNA fragmentation
in traditional semen analysis is likely to provide a more through evaluation
37 Egypt. J. of Appl. Sci., 35 (5) 2020
of semen quality and lead to a better ability to predict male fertility. A
combination of sperm chromatin Dispersion (SCD) test is likely to improve
semen quality assessment and lead to more informed decisions, when
predicting male reproductive efficiency (Enciso et al., 2011) in bull
spermatozoa. In a similar experimental fashion, the SCD test was used to
show the incidence of aneuploidies in spermatozoa containing
fragmemented DNA (Enciso et al., 2011). In addition, sperm DNA
fragmentation has been reported to have a negative effect on fertility and
embryo development (Garcia-Macias et al., 2007) in bull spermatozoa. The
relationship between morphological abnormalities and DNA fragmentation
suggests that the use of SCD for the simultaneous evaluation of sperm
morphology and DNA damage may be useful for the detection of these
defective cells (Enciso et al., 2011).
Table 1 : Copulation time, semen characteristics and DNA
Fragmentation Index of the different semen qualities in the
male dromedary camels (Means±SE).
Items Semen quality
Poor Normal Good
Copulation time (min) 2.65±0.08b 5.16±0.07a 5.74±0.04a
Semen colour Thin Creamy Creamy Creamy
Semen Consistency Semi-viscous Viscous Viscous
Semen-eja. volume (ml) 4.08±0.03b 6.24±0.07a 6.75±0.08a
Hydro. ion concen. (pH) 8.02±0.06a 7.65±0.13a 7.59±0.12a
Sperm motility (%) 28.75±0.29c 57.92±0.36b 75.11±0.89a
Dead spermatozoa (%) 70.68±0.89a 23.86±0.39b 21.94±0.22b
Abnormal spermatozoa (%) 43.51±0.61a 15.14±0.12b 13.52±0.13b
Acrosome damage (%) 16.93±0.18a 7.19±0.18b 5.96±0.11b
Chromatin damage (%) 6.48±0.11a 2.87±0.02b 2.14±0.06b
Sperm-cell concentration (x106/ml) 216.23±8.15c 374.81±11.69b 392.56±2.17a
DNA Fragmentation Index (%) 19.62±0.46a 5.17±0.29b 3.98±0.24b
a-c Within a row, within semen quality, means with different superscripts letters differ
significantly (P<0.05).
3. Sperm mensuration (μm):
Sperm mensuration measurements (μm) of the dromedary camels
was insignificantly affects with different semen qualities of the
dromedary camels (Table 2). Total length of the camel spermatozoa was
insignificantly higher in the camels having Good quality of semen than
both Poor and Normal quality of semen. Similar trends were reported by
Zeidan (1999) in the dromedary camels.
Fertilizing ability of spermatozoa depends on the status of their
shapes and sizes, which affect the course of the acrosomal reaction and
sperm penetration of the ovum (Kondracki et al., 2011). Generally, male
age is one of the significantly causes of variation of spermatozoa
morphology and morphometric dimensions (Banaszewska et al., 2011).
Egypt. J. of Appl. Sci., 35 (5) 2020 38
Table 2 : Effects of semen quality on sperm mensuration (μm)
measurements of the male dromedary camels (Means±SE).
Sperm mensuration (μm) Semen quality Significance
Poor Normal Good
Head of spermatozoa:
Head length 6.02±0.08 6.13±0.12 6.17±0.14 NS
Head width 2.60±0.04 2.81±0.06 2.83±0.05 NS
Head breadth 3.16±0.06 3.25±0.07 3.29±0.11 NS
Head shape index 0.43 0.45 0.45 NS
Tail of spermatozoa
(Mid, Main and End piece):
Tail length 45.73±0.61 45.80±0.68 45.92±0.72 NS
Tail width 1.38±0.02 1.54±0.05 1.57±0.06 NS
Tail breadth 1.69±0.06 1.76±0.08 1.83±0.05 NS
Tail shape index 7.59 7.47 7.44 NS
Total length 51.75±0.74 51.93±0.82 52.09±0.91 NS
Head shape index = Width/length ratio.
Tail shape index = Tail length / Head length ratio.
4. Blood serum constituents:
4.1. Total protein, albumin and globulin concentrations (gm/dI):
Data presented in Table 3 showed that total protein, albumin and
globulin concentrations (gm/dI) in the blood serum were not significant
with different semen qualities of the male dromedary camels. Total
protein, albumin and globulin concentrations of the blood serum were
insignificantly higher in the camels having Normal and Good quality
than Poor semen quality. The decrease of total protein with Poor semen
quality may be attributed to low semen characteristics and adaptability of
camels which represented the potent stimulus of semen quality. Similar
trends were reported by Amin et al. (2007) of the dromedary camels.
4.2. Total cholesterol and testosterone concentrations (mg/100ml):
Total cholesterol and testosterone concentrations (mg/dI) in the blood
serum of the male dromedary camels were significantly (P<0.05) increased
with Normal and Good quality of semen compared to those with Poor
quality of semen (Table 3). The highest (P<0.05) values of the total
cholesterol and testosterone concentrations in the blood serum of the male
dromedary camels were recorded with Good semen quality than either
Normal or Poor semen quality, while the lowest (P<0.05) values were
recorded in the camels having Poor semen quality. Khan and Kohli (1973)
showed higher values in cholesterol concentration of the male camels before
and during rut. It was ranged from 49.8 to 85.3mg/100ml of with a mean
value of 65.58mg/100ml. Moreover, blood serum testosterone of the male
dromedary camels was significantly higher which is parallel to increase of
sexual activity. This reflect the fact that the high androgen in the male camel
with the direct cause of the characteristics of its sexual behavior in the
camels having Normal and Good quality of semen. These results are
partially agreement with those of Bedrak et al. (1983) and Zeidan et
al.(2001) in the male dromedary camel.
39 Egypt. J. of Appl. Sci., 35 (5) 2020
4.3. Mineral concentrations (mg/100ml):
Sodium concentration (mg/100ml) in the blood serum of the male
dromedary camels was significantly (P<0.05) increased in the camel
having Poor quality of semen compared to those with Normal and Good
quality of semen (Table 3). The highest (P<0.05) value of blood sodium
concentration of the male dromedary camels was recorded with Poor
semen quality. While, the lowest (P<0.05) value was recorded with Good
semen quality. These results may be attributed to the combined effects of
both absorption and reabsorption of sodium and chloride from the
alimentary tract and Kidney, under the effect of aldosterone hormone
which had higher levels with Poor quality of semen and this was
accompanied by increase of plasma sodium level (Yagil and Etzion,
1979).
However, Potassium, Calcium and Total phosphorous
concentrations (gm/100ml) and Zinc concentrations (μg/100ml) in the
blood serum of the male camels were significantly (P<0.05) higher of the
camels having Normal and Good quality of semen than Poor quality of
semen (Table 3). The decrease in potassium concentration in the camels
having Poor semen quality may be due to lower semen characteristics
than Normal or Good semen quality of the male dromedary camels.
Similarly, the increase of Calcium, Total phosphorus and Zinc
concentrations may be high feed efficiency and metabolic rate of the
camels having Normal and Good semen quality, consequently high
semen quality compared to those with camels having Poor semen quality
(Table 3). Similar findings were reported by Zeidan and Abbas (2004)
and Amin et al. (2007) of the male dromedary camel.
4.4. Enzymatic activity (U/L):
Activity of AST and ALT enzymes in blood serum of the male
dromedary camels was significantly (P<0.05) higher of the camel having
Poor quality of semen than Normal and Good quality of semen (Table 3).
Similarly, the activity of the blood serum AST and ALT enzymes was
significantly (P<0.05) increased in the camels having Normal semen
quality compared to those with Good semen quality (Table 3). In general,
the blood enzymes are easily and often influenced by the external
condition including feeding practices, type of shelter and many other
aspects of the herd management, since they are ultimately related to
metabolism especially environmental ones when measuring the enzyme
activity in any animal the adaptability of the camels having Normal and
Good semen quality was significantly better than camels with Poor
semen quality. Similar trends were recorded by Zeidan and Abbas
(2004) and Matter (2019).
Egypt. J. of Appl. Sci., 35 (5) 2020 40
Table 3: Effects of different semen quality on blood serum
constituents of the male dromedary camels (Means±SE).
Items Semen quality
Poor Normal Good
Total protein (gm/100ml) 6.84±0.21a 7.03±0.23a 7.52±0.32a
Albumin (gm/100ml) 3.50±0.16a 3.62±0.15a 3.74±0.19a
Globulin (gm/100ml) 3.34±0.15a 3.41±0.19a 3.78±0.21a
Total cholesterol (mg/100ml) 30.72±18.23b 82.35±20.16a 84.13±34.11a
Sodium (mg/100ml) 175.19±6.18a 142.15±5.11b 123.17±3.48c
Potassium (mg/100ml) 1.62±0.13c 11.85±0.38b 13.68±0.42a
Calcium (mg/100ml) 4.12±0.19c 9.43±0.35b 12.16±0.40a
Total phosphorus (mg/100ml) 2.16±0.14c 5.38±0.26b 9.14±0.28a
Zinc (μg/100ml) 87.11±2.14c 113.45±4.23b 128.50±5.18a
Aspartate-aminotransaminase (U/L) 46.12±0.73a 34.60±0.51b 28.35±0.38c
Alanine- aminotransaminase (U/L) 62.13±0.81a 50.22±0.64b 41.53±0.47c
Testosterone concentration (ng/ml) 0.98±0.12c 3.91±0.15b 6.02±0.19a
a-c : Within a rows, within semen quality, means with different superscripts letters
differ significantly (P<0.05).
The second experiment:
1. Percentage of motile camel spermatozoa:
The percentages of motile and storagability of the diluted camel
spermatozoa with LYC extender were significantly (P<0.05)
increased of the camels having Normal and Good semen quality
compared to these with Poor semen quality during storage at 5°C.
Similarly, Good quality of the diluted camel spermatozoa with LYC
extender showed significantly (P<0.05) higher of the percentages of
sperm motility and sperm storagability than Normal quality of
spermatozoa stored at 5°C (Table 4). These changes in sperm motility
and sperm sperm storagability are consistent with the idea that there
is a transition of semen quality from high with Good quality of semen
to low with poor quality of semen. Similar trends were reported by
Zhang et al. (2001) in bull spermatozoa.
It is of interest to note that the prolongation of storage time at 5°C
decreased significantly (P<0.05) percentage of motile camel spermatozoa
(Table 4). In all semen qualities, percentage of motility of the cooled
camel spermatozoa was significantly (P<0.05) maintained during the first
day and then decreased significantly (P<0.05) as time of storage increase.
These results are in agreement with those of Zhang et al. (2001) in bull
spermatozoa. These findings may be due to that the increase of sperm
motility in the first day of storage causes an increase in sperm metabolic
activity, consequently increase of lactic acid production which in turn
exerts a toxic effect of the sperm cells. This phenomenon may be
attributed to decrease in the content of adenosine triphosphate which
activated spermatozoa apparently ability of resynthesizing. This was
accompanied with a precipitous fall in the rate of fructolysis (Mann and
Lutwak-Mann, 1981).
41 Egypt. J. of Appl. Sci., 35 (5) 2020
Table 4 : Mean percentages of motile and storagability of the
dromedary camel spermatozoa during storage at 5°C for up
to 3 days (Means±SE).
Storage time
(days)
Semen quality Mean
Poor Normal Good
0 36.19±0.28 54.82±0.60 68.23±0.74 53.08±0.60a
1 30.25±0.24 49.16±0.52 65.11±0.70 48.17±0.53b
2 21.38±0.18 43.67±0.51 60.24±0.65 41.76±0.50c
3 8.92±0.11 28.14±0.32 45.69±0.52 27.58±0.37d
Overall mean 24.18±0.19C 43.94±0.50B 59.81±0.62A 42.64
Storagability (%) 24.64 54.33 66.96 51.95
A-C Values with different superscripts within a row are significantly different (P<0.05).
a-d Values with different superscripts within a column are significantly different
(P<0.05).
2. Percentage of dead camel spermatozoa:
The percentage of dead camel diluted spermatozoa with LYC
extender was significantly (P<0.05) decreased in the camels having
Normal and Good quality of semen compared to those with Poor quality
of semen during storage at 5°C for 3 days (Table 5). These findings may
be due to bad semen quality, consequently Poor quality of semen.
The prolongation of storage time at 5°C increased significantly
(P<0.05) the percentage of dead camel spermatozoa with all different
semen qualities (Table 5). The percentage of dead spermatozoa increased
significantly (P<0.05) as time of storage increase. Shannon and Curson
(1972) found that dead spermatozoa were a source of amino acid oxidase
which cause the production of H202, consequently, increased the
percentage of dead spermatozoa as the time of storage increase. Similar
trend was reported by Zeidan (2002) in the dromedary camel
spermatozoa.
Table 5 : Mean percentages of dead camel spermatozoa during
storage at 5°C for up to 3 days (Means±SE).
Storage time
(days)
Semen quality Mean
Poor Normal Good
0 61.13±0.72 42.18±0.38 21.87±0.13 41.72±0.39d
1 64.71±0.75 45.76±0.52 24.93±0.17 45.13±0.53c
2 76.18±0.81 50.65±0.63 35.74±0.28 54.19±0.64b
3 87.22±0.93 65.18±0.76 50.16±0.61 67.52±0.84a
Overall mean 72.31±0.82A 50.94±0.65B 33.17±0.26C 52.14
A-C Values with different superscripts within a row are significantly different (P<0.05).
a-d Values with different superscripts within a column are significantly different
(P<0.05).
3. Percentage of abnormal camel spermatozoa:
The percentage of abnormal camel spermatozoa diluted with LYC
extender was significantly (P<0.05) decreased in the camels having
Normal and Good quality of semen compared to those with Poor quality
of semen during storage at 5°C. In addition, the percentage of diluted
Egypt. J. of Appl. Sci., 35 (5) 2020 42
camel spermatozoa with LYC extender showed significantly (P<0.05)
higher in the camel having Good quality than Normal quality of semen
stored at 5°C (Table 6).
It was a very potent and efficient endogenous radical scavenger. It
reacted with the highly toxic hydroxyl radical and provides protection
against oxidative damage to biomolecules. These results are in agreement
with those obtained by Zhang et al. (2001) in bull spermatozoa.
It is of interest to note that the prolongation of storage time at 5°C
increased significantly (P<0.05) the percentage of abnormal camel
spermatozoa with all different semen qualities (Table 6). These results
are in agreement with those of Zhang et al. (2001) in bull spermatozoa.
Table 6 : Mean percentages of abnormal camel spermatozoa during
storage at 5°C for up to 3 days (Means±SE).
Storage time
(days)
Semen quality Mean
Poor Normal Good
0 34.15±0.43 19.23±0.23 8.62±0.09 20.66±0.25d
1 41.26±0.57 20.69±0.26 9.51±0.11 23.82±0.30c
2 53.18±0.61 26.14±0.31 12.36±0.16 30.56±0.40b
3 64.59±0.84 35.92±0.46 17.83±0.18 39.44±0.52a
Overall mean 48.29±0.63A 25.49±0.34B 12.08±0.17C 28.62
A-C Values with different superscripts with a row are significantly different (P<0.05).
a-d Values with different superscripts with a column are significantly different
(P<0.05).
4. Percentage of acrosome damage of spermatozoa:
The percentage of acrosome damage of camel spermatozoa as
affected by semen quality of semen during storage at 5°C was significant
(P<0.05). The highest (P<0.05) value of the percentage of acrosome
damage of spermatozoa was recorded in the camel having Poor quality of
semen, while the lowest (P<0.05) value was recorded in the camels
having Normal and Good quality of semen during storage at 5°C (Table
7).
Jones and Stewart (1979) indicated that extension and cooling of
bull semen to 5°C caused acrosome swelling in about 50% of the
spermatozoa. Subsequent freezing and thawing caused considerable
ultrastructural changes to the acrosomes (disruption of the plasma and
outer acrosome membranes and dispersion of the acrosomal (disruption
of the plasma and outer acrosome membranes and dispersion of the
acrosomal contents) and middle pieces (breakage of the plasma
membrane and a reduction in the electron density of the mitochondrial
matrix) of a high proportion of spermatozoa.
Moreover, storage of semen at low temperatures caused structural
damage as a result of cold shock. The changes involved damage to the
plasma membrane over the acrosome and the outer acrosome membrane
and damage to the plasma membrane of the middle piece. These changes
are followed by a decrease in the proportion of spermatozoa with intact
43 Egypt. J. of Appl. Sci., 35 (5) 2020
acrosomes and an increase in the release of enzymes into the extracellular
medium. Therefore, the morphological characteristics of sperm
acrosomes and enzymes concentration in the extracellular medium with
initial motility gives the best indication so far of initial quality, especially
for frozen bull semen (Zeidan et al., 1998).
The prolongation of storage time at 5°C increased significantly
(P<0.05) the percentage of acrosome damage of the cooled camel
spermatozoa with all different semen qualities during storage at 5°C
(Table 7). These results are in agreement with those of Zhang et al.
(2001) in bull spermatozoa.
Table 7 : Mean percentages of acrosome damage of the camel
spermatozoa during storage at 5°C for up to 3 days
(Means±SE).
Storage time
(days)
Semen quality Mean
Poor Normal Good
0 16.50±0.09 8.23±0.05 3.17±0.02 9.30±0.06d
1 19.73±0.11 10.64±0.07 4.26±0.03 11.54±0.09c
2 28.91±0.17 17.32±0.10 6.15±0.05 17.46±0.12b
3 40.26±0.23 23.54±0.14 10.32±0.08 24.70±0.16a
Overall mean 26.35±0.19A 14.93±0.08B 5.97±0.03C 15.75
A-C Values with different superscripts within a row are significantly different (P<0.05).
a-d Values with different superscripts within a column are significantly different
(P<0.05).
5. Percentage of chromatin damage (%):
The percentage of chromatin damage of the diluted camel
spermatozoa with LYC extender was significantly (P<0.05) lower of
the camels having Normal and Good quality of semen than Poor
quality of semen during storage at 5°C for up to 3 days. Moreover,
the percentage of diluted camel spermatozoa with LYC extender was
significantly (P<0.05) increased of the camels having Good quality of
semen compared to those camels with Normal semen quality (Table
8). Similar trends were recorded by Zeidan et al. (2001) and Matter
(2019) in the dromedary camel spermatozoa.
There are many fluctuations in damage of DNA spermatozoa such
as imperfect of spermatogenesis process, apoptosis, reactive oxygen
species, in vitro handling, type of extender and cryopreservation stress
(Baiee et al., 2017). Lioyd et al. (2012) confirmed that sperm DNA
integrity was better in commercial diluent could be significantly
increased DNA fragmentation during at 5°C for 48 hours. Similar tends
were recorded by Matter (2019) in the dromedary camel spermatozoa.
It is of interest to note that the prolongation of storage time at 5°C
for 3 days was significantly (P<0.05) increased the percentage of
chromatin damage of the camel spermatozoa in all different semen
qualities (Table 8). These results are in agreement with those of Matter
(2019) in the male dromedary camel spermatozoa.
Egypt. J. of Appl. Sci., 35 (5) 2020 44
Table 8 : Mean percentages of chromatin damage of the camel
spermatozoa during storage at 5°C for 3 days (Means±SE).
Storage time
(days)
Semen quality Mean
Poor Normal Good
0 8.12±0.04 4.18±0.02 2.10±0.01 4.80±0.03d
1 10.13±0.05 5.76±0.04 2.89±0.01 6.26±0.04c
2 15.06±0.10 7.14±0.05 5.18±0.03 9.12±0.06b
3 20.72±0.12 11.64±0.06 8.39±0.06 13.58±0.10a
Overall mean 13.50±0.11A 7.18±0.06B 4.64±0.02C 8.44
A-C Values with different superscripts within a row are significantly different (P<0.05).
a-d Values with different superscripts within a column are significantly
different (P<0.05).
6. Enzymatic activity (U/106 spermatozoa):
Activity of AST and ALT enzymes in the camels with Good quality of
semen diluted with LYC extender was significantly (P<0.05) lower than
Normal and Poor quality of semen during storage at 5°C (Tables 9 and 10).
These findings may be explained by the amplified antioxidant enzymes activity,
consequently the ability of seminal plasma to reduce the oxidative stress was
significantly higher with Good quality of semen than Poor quality of semen. In
addition, the enzymatic activity in seminal plasma are good indicator of semen
quality because it measure sperm membrane stability (Corteel, 1980).
Furthermore, many studies have correlated with AST enzyme level in semen
and sperm concentration (Khokhar et al., 1987). Al-Daraji et al. (2002)
confirmed that cell membrane was damaged AST and ALT enzymes released
into the extracellular sperm medium. Graham and Pace (1967) found that
AST and ALT enzymes released into the extracellular medium may reflect the
incubation breakdown of the celluar sperm membrane during storage of semen.
It is of interest to note that the advancement of storage time at 5°C for 3
days increased significantly (P<0.05) the amount of AST and ALT enzymes
released into the extracellular medium in the all semen qualities (Tables 9 and
10). It appears that sperm damage during storage may be associated with
leakage of intracellular enzymes and increased sperm membrane permeability.
Table 9 : Activity of aspartate-aminotransaminase enzyme
(U/106 spermatozoa) of different semen qualities in
the dromedary camel during storage at 5°C for 3 days
(Means±SE).
Storage time
(days)
Semen quality Mean
Poor Normal Good
0 67.18±1.28 49.84±0.91 43.72±0.74 53.58±1.10d
1 73.63±1.39 55.13±1.10 48.57±0.86 59.11±1.23c
2 84.11±2.06 63.72±1.23 54.16±1.12 67.33±1.30a
3 92.43.2.13 74.50±1.42 62.25±1.20 76.39±1.31a
Overall mean 79.33±1.98A 60.89±1.18B 52.17±1.08C 64.10
A-C Values with different superscripts within a row are significantly different (P<0.05).
a-d Values with different superscripts within a column are significantly different
(P<0.05).
45 Egypt. J. of Appl. Sci., 35 (5) 2020
Table 10 : Activity of alanine-aminotransaminase enzyme (U/106
spermatozoa) of different semen qualities in the
dromedary camel during storage at 5°C for 3 days
(Means±SE).
Storage time
(days)
Semen quality Mean
Poor Normal Good
0 53.02±0.83 30.15±0.38 25.34±0.34 36.17±0.61d
1 58.19±0.96 35.24±0.48 29.50±0.36 40.97±0.76c
2 69.14±1.14 42.73±0.51 34.27±0.45 48.71±1.13b
3 85.93±1.98 55.16±1.13 42.65±0.78 61.24±1.18a
Overall mean 66.57±1.27A 40.82±0.75B 32.94±0.42C 46.77
A-C Values with different superscripts within a row are significantly different (P<0.05).
a-d Values with different superscripts within a column are significantly different
(P<0.05).
7. Fertility rate (%):
Conception rates of she-camels artificially inseminated with Poor,
Normal and Good quality of semen during the first day of storage at 5°C
were 11.76, 61.11 and 83.33%, respectively with significantly (P<0.05)
different (Table 11) Fertility rates were significantly (P<0.05) better in
the camels having Good quality than Normal or Poor quality of semen
which may due to high semen characteristics. Similar trends were
reported by Zhang et al. (2001) in bull spermatozoa .
Non of the available literature studied on the effects of semen
quality of the dromedary camels on fertility rates, during storage at 5°C.
Table 11 : Conception rates (%) in the dromedary she-camels
artificially inseminated with different cooled camel semen
qualities.
Semen quality No. of she-camel
inseminated
No. of she-camel
conceived
Conception rate (%)
Poor 17 2 11.76C
Normal 18 11 61.11B
Good 18 15 83.33A
a-c : Within a column, within semen quality, means with different superscripts letters
differ significantly (P<0.05).
In conclusion, copulation time, semen characteristics, blood
constituents, sperm DNA fragmentation Index were better in the camels
with Good quality of semen. In addition, sperm survivability and fertility
rates in the camels having Good quality of semen were significantly better
with Good quality than Normal and Poor quality of semen stored at 5°C.
REFERENCES
Abd El-Raouf, M.F.; M.R. Fatah El-Bab and M.M. Owaida, (1975). Studies
on reproduction in the camel (Camelus dromedaries). V. Morphology
of the testes in relation to age and season. J. Reprod. Fertile., 43: 109-
116.
Abraham, G.E. (1977). Handbook of Radioimmunoassay. Macel Dekker,
Amsterdam, Holland, 5: 591-656.
Egypt. J. of Appl. Sci., 35 (5) 2020 46
Al-Daraji, H.J.; B.T.O.Al-Tikriti and A.A. Al-Rawi (2002). Study of the
trails of endigenous roostert reard during summer months. Iraqian
J. of Agric Sci., 33: 223-278.
Amin, Alia S.A.; Abdoun, K.A. and Abdelatif, A.M. (2007). Seasonal
variation in blood constituents of one-humped camel (Camelus
dromedarius). Pakistan J. Biol. Sci., 10: 1250-1256.
Anouassi, A.; A. Tibary; M. Adnani and A. Sghiri (1994). Preovulatory
phase characterization in Camelus dromedarius and induction of
ovulation. Proc. of Comp. Organized by FIS in Niamy, Niger, pp.
159-167.
Baiee, F.H. ; H. Wahid ; Y. Rosnina ; O.M. Ariff ; N. Yimer ; H. Salman ;
A.A. Tarig and A.M. Khumran (2017). Hypo-osmotic swelling test
modification to enhance cell membrane integrity evaluation in
cryopreserved bull semen. Pakistan J. Tropical Agric. Sci., 40 (2):
257-268.
Banaszewska, D.; S. Kondracki and A. Wysokinska (2011). Effect of age on
the dimensions and shape of spermatozoa of Large White Polish
boars. Arch. Tierz, 54 (5): 504-514.
Banerjee, G.C. (1988). "Feeds and Principles of Animal Nutritions". Mohan
Primalani of Oxford and IBH Publisthing Co. 66. Janpath New Delhi,
India.
Bedrak, E.; Rosenstrauch, A.; Kafka, M. and Friedlander, M. (1983).
Testicular steriodogenesis in the camel (Camelus dromedarius) during
the mating and the non-mating season. Gen. Comp. Endocrinol., 52:
255-271.
Bravo, P.W.; J.A. Skidmore and X.X. Zhao (2000). Reproductive aspects
and storage of semen in camelidae. Anim. Reprod. Sci., 62: 173-193.
Campbell, R.C.; H.M. Dott and T.D. Glover (1956). Nigrosin-Eosin as stain
for differentiating live and dead spermatozoa. J. Agric Sci., 48: 1-8.
Chenoweth, P.J. (2005). Genetic sperm defects. Theriogenology, 64:457-468.
Corteel, J.M. (1980). Effects of seminal plasma on survival and fertility of
spermatozoa kept in vitro. Reprod. Nutr. Dev., 1111-1123.
Duncan, D.B. (1955). Multiple range and multiple F-test. Biometrics. 11: 1-42.
Enciso, M.H. ; S.D.Cisale ; J. Johnston ; Sarasa J.L. Fernandez and J.
Gosalvez (2011). Major morphological sperm abnormalities in the
bull are related to sperm DNA damage. Theriogenology, 76: 23-32
Enciso, M. ; C. Lopez-Fernandez ; J.L. Fernandez ; P. Garcia ; A.
Goselbez and J.Gosalvez (2006). A new method to analyze boar
sperm DNA fragmentation under bright-field or fluoescance
microscopy. Theriognology, 65: 308-316.
Erenpreiss, J.; K.Jepson; A.Giwercman; L.Tsarev; J.Erenpreisa and
M.Spano (2004).Toluidine blue cytometry test for sperm DNA
47 Egypt. J. of Appl. Sci., 35 (5) 2020
conformation: Comparison with the flow cytometric sperm chromatin
structure and TUNEL assys. Hum. Reprod., 19: 2277-2282.
Evenson, D.P. ; K.L. Larson and L.K. Jost (2002). Sperm chro-matin
structure assay. Its clinical use for detecting sperm. DNA
fragmentation in male infertility and comparisons with other
techniques. Intern. J. Andrology, 23: 25-43.
Foote. R.H. and R.W. Bratton (1960). Survival of bovine spermatozoa stored
at 5 and 25°C in extenders containing varying levels of egg yolk,
glucose, glycine. Glycerol, citrate, and other salts. J. Dairy Sci., 43:
1322-1329.
Frenandez, J.L. ; L. Muriel ; M.T. Rivero ; V. Goyanes ; R. Vazquez and
J.G. Alvarez (2003). The sperm chromatin dispersion test a simple
method for the determiation of sperm DNA fragmentaion. Intern. J.
Andrology, 24: 59-66.
Garcia-Macias ;Paulino de Paz Martinez-Pastor ; Alvarez Gomes-Alwes ;
Bernardo Enrique Anel and Luis Anel (2007). DNA
freagmentation assessment by flow cytometry and Sperm-Bos-
Halomax (bright-field microscopy and fluorescence microsopy) in
bull sperm. Intern. J.Andrology, 30: 88-98.
Garnica, J.; R. Achata and P.W.Bravo (1993). Physical and biochemical
characteristics of alpaca semen. Anim. Reprod. Sci., 32: 85-90.
Graham, E.F. and M.M. Pace (1967). Some biochemical changes in
spermatozoa due to freezing. Cryobiology, 4: 75-81.
Hackett, A.J. and J.W. Macpherson (1965). Some staining procedures for
spermatozoa. Canadian Vet. J., 6: 55-62.
Hallap, T. ; S. Nagy ; M. Haard ; U. Jaakma ; A. Johannisson and H.
Rodriguez-Martinez (2005). Sperm chromatin stability in frozen
thawed semen is maintained over age in Al bulls. Theriogenology, 63:
1752-1763.
Jones, R.C. and D.L. Stewart (1979). The effects of cooling to 5 oC and
freezing and thawing on the ultrastructure of bull spermatozoa. J.
Reprod. Fertil., 56: 233-238.
Karras, W. (1952). Waermewasserbad und registratur, ihreentwicklung und
anwendung Deutschland tieraerzlt-ochenschr. 59. (Suppl. 2) 60-62
and 68-69.
Khan, A.A. (1971). Sexual behaviour of the male camel (Camelus
dromedaries) and some studies on semen. MVSc. Thesis, Bikaner
Univ. Undaipur, India.
Khan, A.A. and I.S. Kohli (1973). A note on variations in blood serum
cholesterol in camel (Camelus dromedarius) before and during rut.
Indian J. Anim. Sci., 43:1094-1095.
Khokhar, B.S.; M.Singh and K.C. Chaudhary (1987). Transaminase in
Cattle and buffalso semen in relation to fertility and seminal
Egypt. J. of Appl. Sci., 35 (5) 2020 48
characteristics during moderate and colder seasons. Anim. Reprod.
Sci., 13: 177-182.
Kondracki, S. ; A. Wysokinska ; M.Iwanina ; D.Banaszewska and D.
Sitarz (2011). Effect of sperm concentration in an ejaculate on
morphometric traits of spermatozoa in Duroc boars. Polish. J. Vet.
Sci., 14 (1): 35-40.
Lioyd, R.E.; Fazeli, A.; Watson, P.E. and Holt, W.V. (2012). The oviductal
protein, heat-shock 70-kDa protein 8, improves the longterm survival
of ram spermatozoa during storage at 17 oC in a commercial extender.
Reprod. Fertil. Dev., 24: 543 – 549.
Mann, T. and C. Lutwak-Mann (1981). Male Reproductive Function and
Semen. Springer-Verlag. Berlin, Heidelberg, New York, USA, pp.
264-268.
Matter, M.A.S. (2019). Reproductive and physiological studies on the male
dromedary camels during breeding and non-breeding seasons under
Egyptian condition. MSc. Thesis, Fac. Agric., Al-Azhar University,
Cairo, Egypt.
Murase, T.; K. Okuda and K. Sato (1990). Assessment of bull fertility using
a mucus penetration test and a human chorionic gonadotropin
stimulation test. Theriogenology, 34: 801-812.
Musa, B.; H. Sieme; H. Merkt and B.E.D. Hago (1992). Artificial
insemination in dromedary camels. Proc. 1st Intern. Camel Cong.,
Dubai, U.A.E. pp. 179-182.
Reitman, S. and M. Frankle (1957). A colorimetric method for determination
of semen oxaloacetic and glutamic pyruvic transaminases. Anim.
Clin. Pathol. J., 16: 28-56.
Rodriguez, S. ; V. Goyenes ; E. Segrelles ; M. Blasco ; J. Gosalvez and J.I.
Fernandez (2005). Critically short telomeres are associated with
sperm DNA fragmentation. Fertil. and Steril., 84: 843-845.
Rodriguez- Martinez, H. (2003). Laboratory semen assessment and prdiction
of fertility. Still utopia? Repord in Domestic Anim., 38: 312-318.
Salisbury, G.W.; N.L. Van Demark and J.R. Lodge (1978). Physiology of
Reproduction and Artificial Insemination of Cattle. WH Freeman and
Company, San Francisco, USA.
SAS (2006). SAS Users Guide: Statistical Analysis System. Inst. Inc. Cary, NC,
USA.
Shannon, P. and B. Curson (1972). Toxic effect and action of dead sperm on
diluted bovine semen. J. Dairy Sci., 55: 614-620.
Tietz, N.W. (1982). Fundamentals of Clinical Chemistry. Norbert Saunders
Company, Philadelphia, USA, pp. 401-421.
Watson, P.F. (1975). Use of a giemsa stain to detect changes in acrosomes of
frozen ram spermatozoa. Vet. Rec., 97: 12-15
49 Egypt. J. of Appl. Sci., 35 (5) 2020
Willis, J.W. (1960). The determination of metals in blood serum and tissues by
atamic absorption spertrophotometry. Spactroch. Acta, 16: 259-272.
Wilson, R.T. (1984). The camel. Longman, London, pp. 83-101.
Yagil, R. and Z. Etzion (1979). Seasonal changes in hormones and behaviour
in male camel. Refauh Vet., 36: 70 (Abstr.).
Yassen, A.M. and M.A. El-Kamash (1970). Storagability of buffalo bull
sperm in skim milk extenders. Alexandria, J. Agric. Res., 18: 7-12.
Zeidan, A.E.B. (1999). Effects of age on some reproductive traits of the male
one-humped camels (Camelus dromedarius). Zagazig Vet. J., 27: 126-
133.
Zeidan, A.E.B. (2002). Semen quality, enzymatic activities and penetrating
ability of spermatozoa into she-camel cervical mucus as affected by
caffeine addition. J. Camel Pract. and Res., 9: 153-161.
Zeidan, A.E.B and Abbas, H.E. (2004). Physiological and biochemical
changes in the male dromedary camels during rutting and nonbreeding
seasons. Zagazig Vet. J., 32: 37-48.
Zeidan, A.E.B.; M.N. El-Gaafary and F.E. El-Keraby (1998). Effects of new
packaging method for frozen-bull semen in pellets form on some
biochemical changes and conception rate. Proc. 1st Intern. Conf.
Anim. Prod. Health in Semi-Arid Areas, El-Arish, North Sinai, Egypt,
pp. 223-234.
Zeidan, A.E.B.; A.A.M. Habeeb ; E.A. Ahmadi; H.A. Amer and M.A. Abd
El-Razik (2001). Testicular and physiological changes of the male
dromedary camels in relation to different ages and seasons of the year.
Proc. 2nd Intern. Conf. Anim. Prod. and Health in Semi-Arid Areas,
El-Arish, North Sinai, Egypt., pp. 147-160.
Zhang, B.R.; M. Buhr ; T. Kroetsch and S.P.P. Leibo (2001). Glycine
betaine improves survival of fresh bovine spermatozoa. Reprod. Fertil.
Dev., 13: 187-192.
تأثیر نوعیة السائل المنوی عمى قدرة الحیوانات المنویة عمى البقاء حیه وتحطم
ومعدل الاخصاب فی ذکور الجمال العربیة DNA
علاء زیدان، السید أبو الفتوح أحمدی ، أحمد عامر، ، طارق مهدی ، أحمد لولی دسوقی.
معید بحوث الإنتاج الحیوانی – الدقی – الجیزة – مصر.
اجریت ىذه الد ا رسة عمى عدد 12 ذکر جمل مغربی عند عمر أکبرر مرن 6 حترى 10 سرنوات
عمى تجرربتین. وقرد ترم جمرس السرامل المنرو مرنیم باسرتهدام المیبرل اوصر ناعی. وقرد قسرمت الجمرال
- 40 % ، بیعیررة ≤ 40 - عمررى حسرر حرکررة الحیوانررات المنویررة إلررى ة ةررة مجررامیس )ضررعی ة ≤ 20
00 %( کررذلک تررم تررردیر لترررة الجمررائ، صرر ات السررامل المنررو ، دلیررل تح ررم - %60 ، جیرردة ≤ 60
ابعرراد الحیوانررات المنویررة ومکونررات الرردم لرری ذکررور الجمررال العربیررة )التجربررة ا ولررى(. ولرری ، DNA
وح ظرت عمرى درجرة حر ا ررة LYC التجربرة الةانیرة ترم ته یر نوعیرات السرامل المنرو المهتم رة بمه ر
Egypt. J. of Appl. Sci., 35 (5) 2020 50
°5 م لمردة 3 أیرام )التجربرة الةانیرة(. کرذلک ترم قیراس معردل اوهصرا )%( لمنرو الممرحرة اصر ناعیا
بنوعیات السامل المنو المهتم ة.
وقد أوضحت النتامج أن ىناک تحسن معنرو )عمرى مسرتو 0.05 ( لری لتررة الجمرائ )دقرام (
وحجررم قذلررة السررامل المنررو لرری نوعیررة السررامل المنررو ال بیعیررة والجیرردة عررن نوعیررة السررامل المنررو
الضرعی ة لمجمرال. کران لرون السرامل المنرو کریمری رقیر ، کریمری ، کریمری کمرا کانرت کةالرة السرامل
المنو شبو لزجة ، لزجة ، ل زجة لی نوعیة السامل المنو الضرعی ة ، ال بیعیرة ، الجیردة عمرى التروالی
لری السرامل المنرو وابعراد (pH) لری الجمرال العربیرة. ىرذا بالإضرالة إلرى أن قیمرة ا س الییردروجینی
لی الجمال العربیرة لرم تهتمر معنویرا براهت نوعیرة السرامل المنرو . زیرادة (μm) الحیوانات المنویة
النسربة الممویرة لحرکرة الحیوانررات المنویررة وترکیزىرا ) 610 /مرل( ، بینمرا انه ضرت النسربة الممویرة لکرل
لری DNA مرن الحیوانرات المن ویرة المیترة والشراذة وشرذوذ اوکروسروم وشرذوذ الکرومراتین ودلیرل تح رم
الحیوانرات المنویررة لمجمرال العربیررة معنویررا )عمرى مسررتو 0.05 ( لرری الجمرال ذات النوعیررة ال بیعیررة
والجیردة مرارنرة بنوعیرة السرامل المنرو الضرعی ة. ىرذا بالإضرالة إلرى أن ىنراک زیرادة ریرر معنویرة لری
بینمرا ا زد ، (gm/100ml) ترکیرز کرل مرن البرروتین الکمری ، اولبیرومین والجموبیرولین لری سریرم الردم
معنویرررا )عمرررى مسرررتو 0.05 ( لنوعیرررة السرررامل (mg/100ml) ترکیرررز کرررل مرررن الکولیسرررترول الکمررری
المنو ال بیعیة والجیدة عن نوعیة السامل المنرو الضرعی ة لری الجمرال. مرن ناحیرة أهرر کران ىنراک
ALT, AST ونشرا إنرزیم (mg/100ml) زیادة معنویرة )عمرى مسرتو 0.05 ( لری ترکیرز الصرودیوم
لری الجمرال ذات النوعیرة الضرعی ة مرارنرة بال بیعیرة والجیردة. بالمةرل کران ىنراک زیرادة معنویرة )عمرى
،(mg/100ml) الکالسررریوم ، (mg/100ml) مسرررتو 0.05 ( لررری ترکیرررز کرررل مرررن البوتاسررریوم
لررری الجمرررال (ng/100ml) والتستسرررترون (μg/100ml) الزنرررک ، (mg/100ml) ال وسررر ور الکمررری
ذات نوعیرة السرامل المنرو ال بیعیرة والجیردة عرن نوعیرة السرامل المنرو الضرعی ة )التجربرة ا ولرى(.
LYC زیرادة النسربة الممویرة لکرل مرن حرکرة الحیوانرات المنویرة وقردرتیا عمرى الح رظ والمه رة بمه ر
بدرجررة معنویررة )عمررى مسررتو 0.05 ( ، بینمررا انه ضررت النسرربة الممویررة لکررل مررن الحیوانررات المنویررة
بدرجرة معنویرة )عمرى ALT, AST المیتة والشراذة وشرذوذ ا کروسروم وشرذوذ الکرومراتین ونشرا إنرزیم
مسرتو 0.05 ( لری الجمرال ذات نوعیرة السرامل المنرو ال بیعیرة والجیردة عرن نوعیرة السرامل المنرو
الضعی ة والمح وظة عمى درجة ح ا ررة 5°م )التجربة الةانیرة(. مرس الترردم لری لتر ا رت الح رظ عمرى درجرة
حر ا ررة 5°م انه ضرت نوعیرة السرامل المنرو وذلرک لری نوعیرات السرامل المنرو المهتم رة بدرجرة معنویرة
)عمى مستو 0.05 ( وذلک لی لت ا رت الح ظ المتوالیرة عمرى درجرة حر ا ررة 5°م لمردة 3 أیرام. انه ضرت
نسرربة اوهصررا )%( لرری النررو الممرحررة اصرر ناعیا معنویرراع )عمررى مسررتو 0.05 ( بنوعیررة السررامل
المنو الضعی ة عن نوعیة السامل المنو ال بیعیة والجیدة.
51 Egypt. J. of Appl. Sci., 35 (5) 2020