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Evaluation Of Fertilizing Capacity Of Holstein Friesian Bull Semen Based On Fertility Associated Proteins And In Vitro Characteristics Of Spermatozoa

KrishiKosh

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Title Evaluation Of Fertilizing Capacity Of Holstein Friesian Bull Semen Based On Fertility Associated Proteins And In Vitro Characteristics Of Spermatozoa
 
Creator Krishnan, G.
 
Contributor Thangavel, A.
Loganathasamy, K.
Veerapandian, C.
Kumarasamy, P.
 
Subject Scrotal circumference
Fertility associated proteins
Sperm cell morphology
Plasma and functional membrane integrity
Mitochondrial membrane potential
Lipid peroxidation
 
Description The accurate evaluation of fertility of bulls used for AI is of utmost
importance. Hence, at present attention is being directed towards the assessment of
fertility associated proteins in the seminal plasma and sperm membrane as additional
tool along with breeding soundness examination to evaluate the breeding status of a
bull. In the present study, 17 adult Holstein Friesian breeding bulls maintained at
Central Frozen Semen Production and Training Institute, Hessarghatta, Bangalore
were utilized. The bulls were divided into 8 groups based on presence or absence of
55 kDa, 28 kDa and 24 kDa fertility associated proteins. Scrotal circumference (SC)
measurement was taken transversely at the greatest diameter of the scrotum, using a
scrotal measuring tape. SC was found to be increased positively from 32.33 ± 0.33 to
42.50 ± 0.50 cm as the age increased from 2 to 10 years respectively. Bulls with
higher scrotal circumference of 40.60 ± 1.57 cm (at 7 years) produced greater semen
volume (8.04 ± 0.43 ml). The concentration of sperm cell was recorded between
1,108.37 ± 55.54 to 1,113.91 ± 96.65 million/ml among the bulls positive for fertility
associated proteins. Whereas, bulls negative for fertility associated proteins (group
VIII) had sperm concentration of 1,089.88 ± 136.33 million/ml.
The per cent of sperm cells with progressive motility was significantly
(P < 0.01) higher in the groups with fertility associated proteins in fresh (56.33 ± 3.07
- 62.33 ± 3.24 %) and frozen-thawed semen (49.58 ± 3.40 - 57.67 ± 4.22 %) in
comparison with the group without fertility associated proteins (fresh semen: 37.00 ±
3.19 %; frozen-thawed semen: 30.50 ± 2.22 %). The sperm cell abnormalities was
significantly (P < 0.05) lower (8.88 ± 0.79 - 10.72 ± 1.02 %) in groups with fertility
associated proteins when compared to group VIII (14.47 ± 1.71 %) in fresh semen. In
frozen-thawed semen, sperm cell abnormalities were increased significantly
(P < 0.01) in all the group of bulls where the increase was significantly (P < 0.01)
lower (11.11 ± 0.80 - 12.73 ± 0.99 %) in the bulls with fertility associated proteins in
comparison with the bulls without fertility associated proteins (17.61 ± 2.02 %).
In the present study, the SDS-PAGE of seminal plasma and sperm membrane
protein revealed 13 protein bands with the molecular weight ranging from 6.5 to 205
kDa. The protein bands with molecular weight of 6.5, 14/15, 28, 45, 66 and 116 kDa
were present in all the bulls (100 %) in both seminal plasma and sperm membrane.
The seminal plasma proteins had bands with molecular weights of 26, 36 and 84 kDa
in 88.24 per cent of bulls. Also, 76.47 per cent of the bulls seminal plasma had 55
kDa protein band, whereas sperm membrane had 70.59 per cent of the bulls. The
seminal plasma and sperm membrane of all the bulls (100 %) had a heparin binding
protein with molecular weight of 14/15 kDa. In seminal plasma, the heparin binding
proteins of 24 and 28 kDa were present in 70.59 per cent and 64.71 per cent of the
bulls respectively. In the sperm membrane of 58.82 per cent and 47.06 per cent of the
bulls were positive for 24 kDa and 28 kDa heparin binding protein, respectively.
The per cent of sperm cells with intact plasma membrane was significantly
(P < 0.05) higher (61.43 ± 0.57 to 62.67 ± 0.70 %) in groups with fertility associated
proteins in comparison with the group VIII (58.01 ± 0.43 %) in frozen-thawed semen.
The functional membrane integrity was significantly (P < 0.05) higher in bulls with
fertility associated proteins (83.42 ± 0.83 - 83.67 ± 0.49 %) in comparison with the
groups without fertility associated proteins (75.78 ± 1.02 %) in fresh semen. The per
cent of sperm cells with functional membrane was decreased significantly (P < 0.01)
in the frozen-thawed semen of all the bulls. However, the functional membrane
integrity was significantly (P < 0.05) higher (67.19 ± 1.86 to 71.73 ± 2.61 %) in bulls
with fertility associated proteins when compared to group without fertility associated
proteins (59.88 ± 0.53 %) in frozen-thawed semen.
Bulls positive for fertility associated proteins had significantly (P < 0.05) more
(84.08 ± 0.37 to 84.41 ± 0.52 %) number of sperm cells with intact acrosome than the
bulls without fertility associated proteins (81.78 ± 1.02 %) in fresh semen. The postthaw
acrosome integrity was significantly (P < 0.01) reduced in all the bulls
irrespective of presence or absence of fertility associated proteins. Even though, the
decrease was significantly (P < 0.05) low in (77.16 ± 1.63 to 77.98 ± 1.66 %) in bulls
with fertility associated proteins in comparison with the bulls without fertility
associated proteins (71.73 ± 2.61 %) in frozen-thawed semen.
Bulls with fertility associated proteins had higher (97.84 ± 0.13 to 98.11 ±
0.22 %) per cent of intact sperm nuclear DNA when compared to bulls without
fertility associated proteins (93.67 ± 0.67 %) in fresh semen. Sperm chromatin
integrity was reduced significantly (P < 0.01) in frozen-thawed semen in all the bulls.
However, the decrease was significantly (P < 0.05) low (95.51 ± 0.49 to 94.91 ±
0.27 %) in bulls with fertility associated proteins than the bulls without fertility
associated proteins (90.84 ± 0.63 %).
The sperm cells of the bulls positive for fertility associated proteins traveled
significantly (P < 0.01) more distance (30.54 ± 0.45 to 31.67 ± 0.56 mm) in the
bovine cervical mucus when compared to the bulls negative for fertility associated
proteins (27.00 ± 0.56 mm). The MDA level was significantly (P < 0.01) low (0.382 ±
0.03 to 0.459 ± 0.04 μ mol/ml) in bulls with fertility associated proteins in
comparison with the bulls without fertility associated proteins (0.740 ± 0.05 μ
mol/ml). In the frozen-thawed semen, MDA level was increased significantly
(P < 0.01) in all the bulls. However, the increase was significantly (P < 0.05) lower
(0.638 ±0.02 to 0.833 ±0.12 μ mol/ml) in the bulls with fertility associated proteins
than group VIII (1.445 ±0.19 μ mol/ml).
It is concluded that bulls with larger scrotal circumference produced higher
semen volume. The bulls positive for fertility associated proteins had better semen
quality in comparison with bulls negative for fertility associated proteins. The fertility
associated proteins had preserved the structural and functional integrity of plasma
membrane and organelles during cryopreservation.
 
Date 2016-05-23T16:28:42Z
2016-05-23T16:28:42Z
2013
 
Type Thesis
 
Identifier http://krishikosh.egranth.ac.in/handle/1/66236
 
Language en
 
Format application/pdf
 
Publisher Tamil Nadu Veterinary and Animal Sciences University