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Present address: 1Veterinary Officer, Tamil Nadu. 3,6Senior
Scientist (bhakat.mukesh@gmail.com, gmondal1075
@gmail.com), 2,4,7Principal Scientist (vsrainandri@gmail.com,
tushar@gmail.com, guptaak2009@gmail.com), 5Ph.D Scholar
(drpr06@gmail.com), Artificial Breeding Research Centre.
In India contribution of 20.81% crossbred cows for
improvement of milk production cannot be ignored, but
major constrain has witnessed during use of crossbred bulls,
as poor semen quality, freezability and poor libido are one
of the major reasons of culling in crossbred bulls (Khatun
et al. 2013). To overcome such problems and to maximize
the efficiency of breeding bulls researcher have
administered hormones (Gauthaman et al. 2003, Sieme et
al. 2004 and Ramchander et al. 2004), fed zinc (Osman et
al. 2000, Kendall et al. 2000) and Ayurvedic treatment
(Brown 2000, Giuliano and Allard 2001). Defective
secretion of reproductive hormones leads to lack of libido,
poor semen quality and poor semen freezability in adult
animals. Treatment of bulls with Ayurvedic product and
hormone act through hypothalamic-gonadal-axis to improve
the libido and semen production performance has been
attempted so far by different researchers (Severiano et al.
2007, Baskaran and Dubey 2004). Besides, genetical and
climatic factors one of the major contributor of subfertility
problem in crossbred bulls is mineral deficiency especially
Indian Journal of Animal Sciences 86 (6): 53–00, June 2016/Article
Improvement of sexual behavior and semen quality by therapeutic
approach and Zinc supplementation on Karan Fries
M SABHAPATI1, V S RAINA2, M BHAKAT3, T K MOHANTY4, P R SHIVAHRE5, G MONDAL6 and A K GUPTA7
ICAR-National Dairy Research Institute, Karnal, Haryana 132 001 India
Received: 5 September 2015; Accepted: 20 October 2015
ABSTRACT
Sub-fertility is one of the major concerns in crossbred bulls as 42.98% bulls are getting culled due to sub
fertility problems therefore, a study was planned on 18 Karan Fries bulls producing poor quality semen or not
mounted.They were randomly grouped into 3 treatment groups of 6 animals each and 2 normal bulls were kept as
control. Sexual behavior and semen quality were studied during pre therapy (1 month), therapy (90 days) and post
therapy (90 days) period. Data were analyzed using ANOVA. During therapy period, first group treated with 10 g
of herbal product orally, second group with 0.0082 mg of GnRH intramuscularly at 10 days interval and third group
with 80 ppm of zinc for ninety days for each group. The results depicted that herbal and GnRH treatment significantly
reduced the reaction time, whereas herbal, GnRH treatment and zinc supplementation significantly improved the
libido score and reduced% of non-mounts during the therapy and post-therapy period. The volume of semen and
level of testosterone improved in all treatments groups. Herbal, GnRH treatment and zinc supplementation increased
the average number of good and medium quality ejaculates per month per bull and maximum efficiency achieved
during the treatment period. There was increase in% live sperm and decrease in abnormality in herbal treated and
zinc supplemented group. Therefore, it can be concluded that GnRH therapy, herbal treatment and zinc
supplementation will be effective to overcome the sub fertility problem in crossbred bulls.
Key words: Crossbred bull, GnRH therapy, Herbal treatment, Semen quality, Sexual behavior, Zinc supplementation
zinc (Zn) as it plays an important role in improving male
fertility as zinc is a major component of various enzymes
especially 200 metallo-enzymes, which regulate several
cellular metabolic activities. Researchers (Kendall et al.
2000, Osman et al. 2000) have reported that higher
concentration of zinc has positive impact on semen quality,
may be due to presence of high zinc concentrations in male
accessory sex glands, seminal plasma and spermatozoa.
Therefore, the study was planned to overcome subfertility
problem in crossbred bull by therapeutic intervention using
hormone, Ayurvedic treatment and zinc supplementation.
MATERIALS AND METHODS
The study was carried out at Artificial Breeding Research
centre, ICAR-National Dairy Research Institute, Karnal.
Bulls were kept in loose housing system and provide 1 h
exercise in the bull exerciser, 1 day before semen collection
to maximize the libido of bulls and to ensure quality semen
production (Singh 2014). Karan Fries bulls (18) that are
not fulfilling the criteria of possessing more than 60%
progressive motility; not exceeding more than 20% total
abnormality and 4% primary abnormalities with post thaw
motility of 40% for more than 3 months were selected.
Karan Fries bulls (18) were randomly grouped into 3
treatments of 6 animals each. Experimental period was
54 SABHAPATI ET AL. [Indian Journal of Animal Sciences 86 (6)
x
categorized as pre-therapy, therapy and post therapy. During
therapy period following regimen was followed
(P<0.01) as compared to the pre-therapy and during therapy
period, may be due to summer stress during post therapy
period. Herbal and GnRH treatment significantly (P<0.01)
reduced the reaction time during therapy (29.96 and 41.10
sec) and post therapy period (30.90 and 33.89 sec) as
compared to the pre-therapy period (44.41 and 62.10 sec).
In similar line Hadi (1970) reported the use of Tentex Forte
in Nagpuri Murrah buffaloes and Sahiwal bulls Baskaran
and Dubey (2004) reported that use of Buserelin acetate in
Holstein Friesian and Jersey bulls reduced reaction time.
The decrease in reaction time in case of zinc
supplementation is in consonance with finding reported by
Osman et al. (2000) in buffalo bulls may be due to increased
level of circulatory testosterone, which is also evident after
GnRH treatment (Jimenez et al. 2007).
Herbal treatment, GnRH injection and zinc
supplementation significantly (P<0.01) improved the libido
score from pre-therapy period to therapy period and it
reached highest during post therapy period. This might be
due to the increased testosterone level mediated through
hormonal-gonadal-axis, resulted in decreased number of
mounts before ejaculation (Sieme et al. 2004). The
improvement of reaction time and libido score due to use
of herbal product may be due to androgen like activity of
the herbs as Giuliano and Allard (2001) reported that
presence of protodioscin (Brown 2000) and dopamine in
the herbal product improved the mounting activity and
erection. Not mount frequency (3.13%) encountered while
semen collection in control animals during post therapy
period, but it was not found during pre-therapy period and
during therapy period, may be due to better environmental
condition as compared to post therapy period.
In T2, T3 and T4 group, the not mount frequency was
reduced. Reduction of non mount frequency using herbal
treatment may be due to presence of sexually enhancing
chemicals protodioscin and dopamine in T. terrestris and
Mucuna plants, which stimulated the mounting activity
followed by better erection (Gauthaman et al. 2003).
Whereas, in GnRH treatment and zinc supplementation
group circulating testosterone level increased from 1.58 ng
during pre therapy period to 2.79 ng during therapy period.
In similar line Osman et al. (2000) reported reduction of
testosterone production in case of zinc deficiency.
Treatment No. of Period of Treatments
groups animals observation
Control (T1) 2 7 months Normal fertile bulls
(control)
Herbal (T2) 6 7 months 5 g speman powder + 5 g
Tentex forte daily for 90
days (orally)#
GnRH (T3) 6 7 months 0.0084 mg of Buserelin
acetate (Synthetic
GnRH)* for 90 days at 7
days interval (Intra
muscularly after semen
collection)
Zinc (T4) 6 7 months 80 PPM extra zinc
feeding daily for 90 days
(In feed)#
* Gynarich Intra muscular injection immediately after the end
of semen and blood collection at weekly intervals; # inorganic
zinc (zinc sulphate) and ayurvedic powder mix were given along
with concentrate mixture for 90 days.
Immediately after semen collection, semen was
evaluated for volume, sperm concentration
(hemocytometer), mass activity, eosin-nigrosin staining and
HOST. Sexual behavior parameters like reaction time, libido
score and per cent non mount activity were evaluated as
per Anzar et al. (1993).
Statistical analysis: The per cent of semen parameter
and other different parameters were calculated by proportion
using descriptive statistics. The semen production records
were subjected to statistical analysis using ANOVA. Group
comparison, percentage data arcsine transformations were
also done for data analysis (Snedecor and Cochran 1994).
RESULTS AND DISCUSSION
Improvement of semen quality and sexual behaviour
after Zinc supplementation, Ayurvedic and GnRH treatment
are presented in Tables 1, 2.
Sexual behavior: In T1 group, post therapy period
average reaction time (38.57 sec) was significantly higher
Table 1. Mean± SE values of sexual behavior during different therapy periods
Treatments Therapy period
Pre During Post
Reaction Libido % Non Reaction Libido % Non Reaction Libido % Non
time score mount time score mount time score mount
(Seconds) activity (Seconds) activity (Seconds) activity
Control (T1) 22.97 b ±4.62 8.67 a ±0.33 0 26.69 b ±1.64 9.18 a ±0.38 0 38.57 a ±5.87 7.70 b ±1.16 3.13
Herbal (T2) 44.41 a ±1.42 4.33 b ±0.50 16.67 29.96 b ±1. 90 6.60 a ±0.20 0.81 30.99 b ±6.36 6.47 a ±0.97 0
GnRH (T3) 62.10 a ±19.75 3.33 b ±0.76 33.33 41.10 a b ±6.34 5.21 a ±0.44 8.11 33.89 b ±10.21 6.14 a ±1.64 2.33
Zinc (T4) 32.63±8.97 2.33 b ±0.71 46.15 27.67±2.81 4.50 a ±0.45 7.77 33.13±12.43 4.87 a ±2.74 7.23
Mean bearing different superscript within row differ significantly (*P<0.05).
June 2016] THERAPEUTIC APPROACH AND ZINC SUPPLEMENTATION IN BREEDING BULLS 55
x
Seminal characteristics
The semen quality recorded for different treatment
groups in 3 different therapy periods are presented in Table
2. In control group volume, mass activity, individual
motility, sperm concentration, acrosome integrity and HOST
were nonsignificant (P>0.05) during 3 treatment periods
whereas the% live sperm count was significantly reduced
during therapy period and post therapy period as compared
to pre-therapy period. In herbal treatment group highest
individual motility, concentration, live per cent sperms,
acrosome integrity and HOST% was recorded during
therapy period. The decline in the values during post therapy
period for above parameters may be due to withdrawal of
herbal treatment. The improvement in semen quality may
be due to improved spermatogenesis functions of testes and
accessory sex gland like prostate and seminal vesicles along
with improvement of libido as reported by Pardanani et al.
(1976) after treatment with speman powder in oligospermic
men.
In GnRH treatment group all semen quality parameters
improved. The results are in agreement with Sieme et al.
(2004) and Ramchander et al. (2004) who also reported
significant improvement in semen volume achieved by
GnRH treatment. The improvement of semen quality may
be due to stimulatory effect of GnRH leads to release of
FSH and LH from anterior pituitary. LH act on leydig cells
to synthesize testosterone, which inturn improves the
functions of all sex glands. There is also a school of thought
that GnRH treatment restores seminiferous tubule function
either through FSH and LH or FSH and testosterone through
synthesis of androgen binding protein and secretion of
tubular fluid. In case of sperm concentration contradictory
finding was reported by Bhaskaran and Dubey (2005) may
be due to the variation in GnRH dose.
All seminal parameters were increased during zinc
supplementation except concentration (Table 2). The
experiment result was in compliance with the earlier
findings of Kendall et al. (2000). The increase in semen
volume after zinc supplementation may be due to
improvement of secretory function of accessory sex glands
as Zn stimulates spermatogenesis, primary and secondary
sex organ’s growth and development and function of
prostate gland in various species (Kumar et al. 2014). Zinc
supplementation increased the mass activity and% motile
spermatozoa during supplementation which was in
correspondence with the findings of Osman et al. (2000)
and Kumar et al. (2006). Zinc is an important element for
tail morphology and sperm motility and it is involved in
the catabolism of lipids in the sperm mid-piece that is the
principal source of energy required for the movement of
spermatozoa. Zinc supplementation significantly improved
(P<0.01) mean% of live sperm count during and post-
therapy periods as compared to the pre-therapy period. The
results are in consonance with Kumar et al. (2014). The
improvement in livability and membrane integrity is due
reduction of oxidative stress through production of
antioxidative enzymes, superoxide dismutase and
Table 2 Mean ± SE values of semen quality during different therapy periods in KF bulls
Treatment Therapy Period
Pre therapy During therapy Post therapy
Volume Mass Individual Conc. live Acrosome HOST Volume Mass Individual Conc. live Acrosome HOST Volume Mass Individual Conc. live Acrosome HOST
activity motility integrity activity motility integrity activity motility integrity
Control 4.00 2.44 48.75 925.00 88.00 a 85.88 47.25 3.78 2.36 48.33 791.03 82.54 b 84.10 46.13 3.65 2.55 49.68 864.52 83.07 b 84.29 48.35
(T1) ±0.37 ±0.11 ±2.80 ±44.32 ±0.96 ±1.20 ±1.10 ±0.22 ±0.13 ±2.45 ±43.54 ±0.92 ±0.84 ±1.56 ±0.21 ±0.11 ±2.65 ±32.971 ±0.71 ±0.84 ±1.40
Herbal 4.23 1.87 35.33 626.67 67.91 b 79.60 a b 37.93 4.66 1.79 37.38 666.80 79.40 a 82.26 a 40.19 5.06 1.58 31.75 589.58 76.43 a 78.71 b 35.88
(T2) ±0.38 ±0.21 ±4.15 ±58.73 ±1.88 ±1.42 ±2.07 ±0.17 ±0.08 ±1.67 ±25.31 ±0.84 ±0.57 ±0.96 ±0.18 ±0.10 ±1.97 ±29.921 ±0.92 ±0.63 ±1.03
GnRH 3.45 b 1.55 33.00 630.00 71.58 79.70 36.20 4.60 a 1.75 36.23 657.97 78.87 80.06 39.04 4.88 a 1.80 36.38 667.72 76.91 79.81 39.01
(T3) ±0.52 ±0.20 ±4.36 ±65.49 ±3.71 ±1.59 ±2.32 ±0.21 ±0.11 ±2.14 ±32.46 ±1.44 ±0.79 ±1.29 ±0.20 ±0.11 ±2.24 ±33.012 ±1.04 ±0.77 ±1.29
Zinc 3.93 1.71 a 35.00 a 685.71 a 59.41 c 79.14 a 37.00 b 4.39 1.84 a 37.79 a 682.63 a 81.73 b 80.75 a 40.13 a 4.88 1.36 b 28.20 b 549.33 b 75.21 a 76.61 b 33.89 b
(T4) ±0.70 ±0.24 ±4.76 ±64.29 ±2.227 ±1.63 ±2.42 ±0.164 ±0.11 ±2.07 ±31.88 ±1.06 ±0.77 ±1.24 ±0.19 ±0.11 ±2.269 ±33.71 ±1.23 ±0.77 ±1.26
Mean bearing different superscript within row differ significantly (*P<0.05).
56 SABHAPATI ET AL. [Indian Journal of Animal Sciences 86 (6)
x
glutathione peroxidase as Zinc act as co factor for the
production of antioxidative enzymes.
Zinc supplementation did not show significant
improvement (p>0.05) in sperm concentration, but
maintained the pre-therapy sperm concentration without
further decline during therapy period. However, withdrawal
of zinc supplementation significantly reduced the sperm
concentration in post-therapy period might be due to
disruption of spermatogenesis process.
SUMMARY
Sub-fertility problem in crossbred bulls is one of the
major concern of semen bank as there is ever increasing
demand of good quality semen of crossbred bulls. Therefore,
to overcome subfertility problem to harvest better quality
semen; herbal treatment, GnRH injection and zinc
supplementation will be effective to improve sexual
behavior and semen quality. However, there is need of better
understanding of the basic physiological role of herbal,
GnRH and zinc on male reproduction after therapeutic
intervention for better utilization of any one of the product
to achieve better result.
ACKNOWLEDGEMENT
The authors are thankful to the Director cum Vice-
Chancellor of National Dairy Research Institute, Karnal for
providing the facilities.
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