ABSTRACT An experiment was conducted with 24 healthy cyclic cows between 2 nd to 5 th lactation by under intensive system of rearing. The animals were randomly divided into Control (C) and Experiment (E) groups with 12 animals in each The control and experiment groups were again further subdivided in groups C 1 and C 2 &amp; E 1 and E 2 comprising six in each. Onset of oestrus was considered as day 0 for superovulatory treatment. The animals of groups C 1 and C 2 &amp; E 1 and E 2 were injected with 5 ml NSS (i/ml) &amp; 21μg GnRH (i/m) (Buserelin acetate) on day 8 of the oestrous cycle, respectively. Each of the experimental animals of groups C 1 and E 1 was superovulated with 400 mg FSH (i/m) in divided and equal doses at 12 h intervals for four days on day 10 to 13 of the oestrous cycle and consequently, each of the experimental animals of groups C 2 and E 2 was also superovulated with 2000 I.U.PMSG injection (i/m) as single dose on day 10 of the oestrous cycle. PGF 2 α 2ml was injected (i/m) at morning time on day 12 of oestrous cycle. At the superovulatory oestrus, all the animals were bred three times at 12 h intervals using frozen semen. The animals were examined per rectally on day 6 of the induced oestrus after first insemination to detect the superovulatory response. Flushing was performed on day 7 of induced oestrus to retrieve embryos from the superovulated cows. In the present experiment, superovulatory response in GnRH treated groups of cows (groups E 1 and E 2 ) were found to be better as compared to that of the animals of control groups but the duration of induced oestrus (h) recorded in groups C 2 (49.02 ± 0.07 h) and E 2 (50.40 ± 0.27 h) were found to be significantly higher (P&lt;0.05) than the value recorded in groups C 1 and E 1 

C Barman

G Kadirvel

P K Bharti

S Doley

S Kumar

V Singh

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111 June 2014  ⎜  Volume 27  ⎜  Issue 1
Indian Journal of Hill Farming
Effect of Presuperovulatory Treatment of Buserelin on
Superovulatory Response and Embryo Recovery in Cattle under
Subtropical Conditions of Meghalaya
S. KUMAR1*, C. BARMAN1, S. DOLEY1, G. KADIRVEL1, P.K. BHARTI1, V. SINGH2
Received 23.5.2014, Revised 15.6.2014, Accepted 28.6.2014
ABSTRACT
An experiment was conducted with 24 healthy cyclic cows between 2nd to 5th lactation by under
intensive system of rearing. The animals were randomly divided into Control (C) and Experiment (E)
groups with 12 animals in each The control and experiment groups were again further subdivided in
groups C1 and C2 & E1 and E2 comprising six in each. Onset of oestrus was considered as day 0 for
superovulatory treatment. The animals of groups C1 and C2 & E1 and E2 were injected with 5 ml NSS
(i/ml) & 21μg GnRH (i/m) (Buserelin acetate) on day 8 of the oestrous cycle, respectively. Each of
the experimental animals of groups C1and E1 was superovulated with 400 mg FSH (i/m) in divided
and equal doses at 12 h intervals for four days on day 10 to 13 of the oestrous cycle and consequently,
each of the experimental animals of groups C2 and E2 was also superovulated with 2000 I.U.PMSG
injection (i/m) as single dose on day 10 of the oestrous cycle. PGF2α 2ml was injected (i/m) at morning
time on day 12 of oestrous cycle.  At the superovulatory oestrus, all the animals were bred three times
at 12 h intervals using frozen semen. The animals were examined per rectally on day 6 of the induced
oestrus after first insemination to detect the superovulatory response. Flushing was performed on day
7 of induced oestrus to retrieve embryos from the superovulated cows. In the present experiment,
superovulatory response in GnRH treated groups of cows (groups E1 and E2) were found to be better
as compared to that of the animals of control groups but the duration of induced oestrus (h) recorded
in groups C2 (49.02
 ± 0.07 h) and E2 (50.40
 ± 0.27 h) were found to be significantly higher (P<0.05)
than the value recorded in groups C1 and E1.
Keywords: Follicle Stimulating Hormone, GnRH, PMSG
1Division of Livestock Production, ICAR Research Complex for NEH Region, Umroi Road, Umiam-793103, Meghalaya
2Dept. of Animal Reproduction, Gynaecology & Obstetrics,  College of Veterinary Sciences & A.H, DUVASU, Mathura-281001, U.P
*Corresponding author’s E-mail: suresh_vet079@rediffmail.com
INTRODUCTION
Superovulation is a key step in the embryo
transfer technology in cattle and requires
administration of a gonadotropin preparation that
mimics the effect of follicle stimulating hormone
(FSH). It is an important and integral part of the
fast developing multiple ovulation and embryo
transfer (MOET) programmes. Superovulation is
done by application of gonadotropin like porcine
pituitary FSH (Barman et al. 2012) with or without
luteinizing hormone in mithun cows or pregnant
mare serum gonadotropin (Saumande and Chupin
1981) with or without luteinizing hormone in cattle.
The success of embryo transfer technique depends
largely on the reliable method for the induction of
superovulation to harvest maximum number of
normal fertile ova or embryos. Although various
advances have been achieved in superovulation
technique, it still remains a weak link in the chain
of events influencing the final outcome of embryo
transfer in cattle. Beneficial effects of
prostaglandins as well as GnRH have been well
documented and are now being used for
improvement of reproductive efficiency in large
ruminants (Kumar et al. 2010). Therefore, the
present study was conducted to determine the effect
of presuperovulatory treatment of buserelin on
Indian Journal of Hill Farming 27(1):111-114 Available online at www.kiran.nic.in
112 June 2014  ⎜  Volume 27  ⎜  Issue 1
Indian Journal of Hill Farming
superovulatory response and embryo recovery in
cattle under subtropical conditions of Meghalaya
in order to understand their reproductive physiology.
MATERIALS AND METHODS
The present experiment was conducted at dairy
unit, ICAR Research Complex for NEH Region,
Umiam, Meghalaya. Twenty four healthy cyclic
cows between 2nd and 5th lactation, free from genital
abnormalities on per rectal examination, having no
previous history of infectious diseases were selected
for the present experiment. Experimental animals
were maintained under intensive system of rearing
and they were examined regularly to ascertain their
health condition and strict hygienic measures were
taken in the shed. The experimental animals were
divided in two groups viz. Control (C) and
Experiment (E) comprising twelve in each. The
control group was again further subdivided in
groups C1 and C2 comprising six in each and
similarly experiment group (E) was also further
subdivided into E1 and E2 comprising six in each.
Onset of oestrus was considered as day 0 for
superovulatory treatment. The animals of groups
C1 and C2 & E1 and E2 were injected with 5 ml NSS
(i/ml) & 21μg GnRH (i/m) (Buserelin acetate) on
day 8 of the oestrous cycle, respectively. Each of
the experimental animals of groups C1and E1 was
superovulated with 400 mg FSH (i/m) (Folltropin,
Veterpharma Inc, Canada) in divided and equal
doses at 12 h intervals for four days on day 10 to
13 of the oestrous cycle. The treatment schedule
for superovulation with FSH 400 mg was as 50/50,
50/50, 50/50 and 50/50 mg respectively and
consequently, each of the experimental animals of
group C2 and E2 was also superovulated with 2000
I.U.PMSG injection (i/m) (Folligon, Intervet
International Gmbh, Germany) as single dose on
day 10 of the oestrous cycle. PGF2á 2ml
(Cloprostenol, Intervet International Gmbh,
Germany) was injected (i/m) at morning time on
day 12 of oestrous cycle.  At the superovulatory
oestrus, all the animals were bred three times at 12
h intervals using frozen semen. The animals were
examined per rectally on day 6 of the induced
oestrus after first insemination to detect the
superovulatory response. Flushing of uterine horns
for non-surgical embryo collection using modified
Dulbecco’s phosphate buffer saline (mDPBS) with
1% heat treated oestrus cow serum was performed
on day 7 of induced oestrus to retrieve embryos
from the superovulated cows according to the
method described by Neto et al. (2005). Data
obtained from the experiment were statistically
analyzed as per the method described by Snedecor
and Cochran (2007).
RESULTS AND DISCUSSION
The results of the present experiment are being
presented in Table 1 and 2. Animals responding to
superovulatory treatment in groups C1, C2, E1 and
E2 were 100%, 83.33%, 100% and 83.33%,
Table 1: Effect of presuperovulatory treatment of buserelin on superovulatory response and embryo
recovery in cattle
Treatment groups No. of Oestrus Mean Total Viable Anovulatory Degenerated Unfertilized
animals response ovulation embryos embryos follicles embryos ova
(%) rate recovery (Mean ± SE) (Mean ± SE) (Mean ± SE)  (Mean ± SE)
(Mean ± SE) (Mean ± SE)
C1
(5 ml NSS+FSH: 6 100.00   6.83NS±1.01  4.17NS±0.91    2.5NS±0.80   0.67NS±0.49   0.67NS±0.49  1.0NS±0.44
400 mg)
C2
(5 ml NSS+PMSG 6 83.33  5.33NS±1.11  1.83NS±0.79   1.67NS±0.65   1.83NS±0.79   0.17NS±0.16   0.5NS±0.49
2000 I.U.)
E1
(21 μg Buserelin 6 100.00  8.17NS±1.30  4.83NS±1.22   2.33NS±0.20    0.67NS±0.20   1.67NS±0.40  0.15NS±0.16
acetate + FSH:
400 mg)
E2
(21 μg Buserelin  6 83.33   5.5NS±1.06   2.83NS±1.64   1.83NS±1.16   0.83NS±0.30   0.83NS±0.54 0.17NS±0.16
acetate +PMSG
 2000 I.U.)
 NS: Non-significant
113 June 2014  ⎜  Volume 27  ⎜  Issue 1
Indian Journal of Hill Farming
respectively. 100 percent oestrus response was
observed in animals of groups C1 and E1.These
findings were in close agreement with the earlier
report of Barman et al. (2012). However, the report
of Mathur et al. (2006) on oestrus response
following superovulation in Frieswal cows was 75
per cent only.
In the present experiment, superovulatory
response in GnRH treated groups of cows were
found to be better as evidenced by apparently  lower
number of palpable follicles as well as apparently
higher numbers of corpora lutea as compared to
that of the animals of control groups. This might
be due to increased growth and maturation of
follicles by combined effect of GnRH and FSH/
PMSG (Nilchuen et al. 2011) or due to follicle
luteinisation and possible ovulation followed by the
emergence of new follicular wave (Schmidt et al.
1996) or also due to the involvement of some factors
including progesterone profile of donor at the time
of superovulation, increased number of follicular
population and endogenous hormonal milieu (Arora
et al. 1996).
The numbers of unovulated follicles obtained
in animals of group C2 were apparently higher as
compared to groups C1, E1 and E2 but analysis of
variance revealed non-significant differences in
respect of anovulatory follicles between GnRH
(buserelin acetate) treated and control groups. This
might be due to insufficient endogenous LH surge
(Taneja et al. 1988).
The number of embryos recovered were found
to be apparently higher in animals of group E1 (4.83
± 1.22) which might be due to increased growth
and maturation of follicles caused by combined
effect of GnRH (Buserelin acetate) and FSH and
also due to genetic variation (Nilchuen et al. 2011)
but statistically found to be non-significant (P>0.05)
between the treated and control groups.
The duration of induced oestrus (h) recorded in
the present experiment were as 43.05 ± 1.15, 49.02
± 0.07, 44.25 ± 0.03 and 50.40 ± 0.27, respectively
for groups C1, C2, E1 and E2. Although apparently
higher value was observed in group E1 as compared
to the value recorded in group C1 but analysis of
variance revealed no significant difference (P>0.05)
between groups C1 and E1. Similar findings have
been also recorded by Bhuyan et al. (2006). The
value recorded in group E2 were found to be
apparently higher than the value recorded in group
C2 but no significant differences (P>0.05) were
found between the two respective groups in respect
of duration of induced oestrus. The present findings
were in close agreement with the earlier report of
Abdoon et al. (2000). The duration of induced
oestrus (h) recorded in groups C2 and E2 were found
to be significantly higher (P<0.05) than the value
recorded in groups C1 and E1. This might be due to
variations in duration of oestrus might be influenced
by the method of treatment, dose, age, breed,
season, cyclicity and stages of reproduction
(Mughal et al. 1998) or could be attributed to
Table 2: Effect of presuperovulatory treatment of buserelin on occurrence of induced and natural oestrus
in cattle
Treatment groups No. of Oestrus Duration of Interval from Duration of
animals response (%) induced induced oestrus natural oestrus
oestrus (hrs.) to the onset of (hrs.)
(Mean±SE) subsequent natural (Mean±SE)
oestrus (days)
(Mean±SE)
C1
(5 ml NSS+FSH: 400 mg) 6 100.00 43.05a±1.15 20.02NS±0.68 24.85NS±0.03
C2
(5 ml NSS+PMSG 2000 I.U.) 6 83.33 49.02b± 0.07 20.15NS±0.15 26.06NS±0.22
E1
(21 μg Buserelin acetate + FSH: 6 100.00 44.25ac±0.03 22.5NS±1.38 25.50NS±0.25
400 mg)
E2
(21 μg Buserelin acetate +PMSG 6 83.33 50.40b± 0.27 21.08NS±0.27 27.50NS±0.10
2000 I.U.)
Means with different superscripts in a column differ significantly (P<0.05), NS-non significant
114 June 2014  ⎜  Volume 27  ⎜  Issue 1
Indian Journal of Hill Farming
increased follicular growth caused by hormone
pregnant mare serum gonadotropin (Abdoon et al.
2000).
The mean time intervals from induced oestrus
to the onset of subsequent natural oestrus in the
present experiment were 20.02 ± 0.68, 20.15 ± 0.15,
22.5 ± 1.38 and 21.08 ± 0.27 days, respectively for
groups C1, C2, E1 and E2. Although analysis of
variance revealed no significant differences for time
intervals recorded from induced oestrus to the onset
of subsequent natural oestrus among the groups C1,
C2, E1 and E2 but apparently longer time interval
was recorded in group E1 (22.5
 ± 1.38 days)
followed by groups E2, C2 and C1. This may be
attributed to elevated concentrations of circulating
P4 due to more numbers of CL. The high
concentration of P4 exerts negative feedback effect
on LH secretion; thereby causing the attenuation
of follicular growth (Barman et al. 2012).
The mean duration of subsequent natural oestrus
(h) recorded in the present experiment were 24.85
± 0.03, 26.06 ± 0.22, 25.50 ± 0.25 and 27.50 ± 0.10
h, respectively for groups C1, C2, E1 and E2.
Although apparently higher values were observed
in group E2 followed by groups C2, E1 and C1 but
analysis of variance revealed no significant
differences for duration of subsequent natural
oestrus among different groups. The present
findings recorded in respect of duration of
subsequent natural oestrus were in close agreement
with the earlier report of Bhuyan et al. (2006).
ACKNOWLEDGEMENT
The authors wish to acknowledge the Director,
ICAR Research Complex for NEH Region, Umiam,
Meghalaya, for providing all necessary
infrastructure facilities for carrying out the present
experiment.
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Effect of Presuperovulatory Treatment of Buserelin on Superovulatory Response and Embryo Recovery in Cattle under Subtropical Conditions of Meghalaya

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Effect of Presuperovulatory Treatment of Buserelin on Superovulatory Response and Embryo Recovery in Cattle under Subtropical Conditions of Meghalaya

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