Vol 7 No 1, p 83-86

EFFECTS OF DIFFERENT ESTROUS SYNCHRONIZATION PROTOCOLS ON ESTRUS AND

SUBSEQUENT FERTILITY IN CYCLING COWS
Amit Kumar*, JS Mehta, Swati Ruhil and GN Purohit
College Of Veterinary and Animal Science, RAJUVAS, Bikaner- 334 001, Rajasthan, INDIA
Department of Veterinary Gynaecology and Obstetrics
*dr.amit1172@gmail.com
ABSTRACT
Present study was conducted on 53 healthy cyclic cow of 2-5 parity and >90 day postpartum to evaluate the
efficacy and subsequent fertility of three estrus synchronization protocols i.e. CIDR-PG (n=13), Ovsynch (n=15), Heatsynch
(n=15). The animals of control group (n=10) remained untreated. After end of the protocol all the treated cows were
observed daily morning-evening for 7 days for the occurrence of signs of estrus and inseminated artificially 12 to 18 hrs after
the onset of estrus. Cohort development of developing follicles (5 mm) was monitored by ultrasonography and revealed
that mean total no. of follicles on the left, right and both the ovaries recorded were not significantly different between the
protocols. The mean diameter of the dominant follicle showed significant difference between the groups. Mean duration of
onset of estrus post protocol was shortest in ovsynch protocol (27.46±0.97 h) whereas mean duration of estrus was
significantly longer in CIDR-PG protocol (23.53±0.96 h). All the inseminated but not returned cows were scanned
ultasonographicaly or examined manually through rectum for the confirmation of pregnancy at 35 and 60 days of
pregnancy and revealed 60 conception rates in the heatsynch and ovsynch protocol whereas 54.54 in CIDR-PG based
protocol. It was concluded that among these estrous synchronization protocol heatsynch protocol was most effective for
induction of estrus and expression of signs of estrus while ovsynch protocol as most effective in term of conception rate.
Keywords: CIDR, estrous synchronization, Heatsynch, Ovsynch.

Combination of GnRH with the Prostaglandin F2
alpha (Prusley et al, 1995) and Progesterone (Patterson
et al, 1999) based synchronization programme has shown
a new direction in the estrus synchronization of cattle
with the manipulation of follicular development (Shukla
and Garg, 2012 ). Many of these estrus synchronization
protocols are not only able to synchronize the growth of
dominant follicles but also the time of luteal regression.
These protocols are very promising and have the potential
to enhance pregnancy rates and the success of artificial
insemination programs (Mahendran et al, 2001; Dudi
et al, 2017). This study was conducted to evaluate the
efficacy and subsequent fertility of three different estrous
synchronization protocols in cyclic cows.
Materials and Methods
A total 53 healthy cyclic cows of 2-5 parity and
>90 days postpartum were subjected to the three estrus
synchronization protocols i.e CIDR-PG (n=13), Ovsynch
(n=15), Heatsynch (n=15) and ten as untreated control
(C). All the cows had regular supplementation of vitamins
and mineral mixture for couple of weeks before the start
of protocols. In CIDR-PG group, EAZI Breed CIDR
(Pfizer limited, India) contained 1.38 g of progesterone
was inserted in all the cows irrespective of the stage of
estrous cycle for 7 days and administered cloprostenol
Ruminant Science
0.5 mg IM (Pragma, Intas Pharma) 1 day before
withdrawal of CIDR. In Ovsynch group, all cyclic cows
(n=15) were randomly administered buserelin 20 µg IM
(Receptal, Intervet, Holland) and considered as day 0.
On day 7th cloprostenol 0.5 mg (Pragma, Intas Pharma)
was given IM and again on day 9th second Receptal 20
µg was given IM. In Heatsynch protocol cyclic cows
(n=15) were given buserelin 20 µg IM and considered
day 0. On day 7th cloprostenol 0.5 mg was given IM and
on day 8th estrdiole velarate 10 mg (Progynon depot)
was given IM. Ultrasonography was done to record the
growth and number of follicles. After this, all the treated
cows were observed daily morning-evening for 7 days for
the occurrence of signs of estrus. Ten healthy, normal cyclic
animals of control group were remained untreated. These
animals were kept for spontaneous estrus by observing at
least 2-3 estrus cycles. After the onset of estrus, all the
cows were checked and parameters like duration of onset
of estrus post protocol, total duration of estrus and the
signs of estrus like vaginal edema, amount of mucous
discharge, restlessness and mounting, bellowing and
tonicity of estrus were recorded. All animals evidencing
estrus were inseminated with frozen semen of high fertility.
All the inseminated but not returned cows were scanned
ultasonographicaly or examined through-rectally for the
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confirmation of pregnancy at 35 and 60 days of pregnancy. Parameters and expressing signs of induced estrus
Statistical analysis The three synchronization protocols had different
The data were analuzed using standard statistical durations of their course as either 7, 8 or 9 days. However,
procedure (Snedecor and Cochran, 1994). when the onset of estrus was considered just from the
end of the protocol, the mean duration was shortest in
Results and Discussion ovsynch and it was highly significantly shortest then rest
Cohort development of developing follicles two protocols (Table 2). Whereas, in the CIDR based
Cohort development of developing follicles (5 mm) protocol and heatsynch protocol the mean duration were
was monitored by ultrasonography and revealed the mean non significantly different with each other. This variation
total No. of follicles on the left, right and both the ovaries can be understood as 2 nd injection GnRH in ovsynch
recorded were not significantly different between the protocol not only hastens the folliculogenesis but also
the ovulatory process and thus this mean duration
protocols (Table 1). The mean diameter of the dominant
recorded was the shortest. Whereas in heatsynch protocol
follicle showed significant difference between the groups.
the last shot of injection estrogen must also have hasten
Heatsynch and ovsynch protocols recorded significantly
the onset of estrus but this significantly longer duration
(P<0.01) higher mean values than control and CIDR-
is because of this duration was counted from the end of
PG based protocol. These significantly higher values could
the protocol. Cavalieri et al (2005) also reported decreased
probably be resulted due to presence of second shot of
interval to estrus by using estradiol injection after the
GnRH or estradiol in the last phase of the protocol PGF2 alpha injection. In heatsynch, the protocol ended
enhanced follicular development bit faster than the other on 8th day whereas in ovsynch the protocol ended on 9th
groups. The exogenous estradiol had just supplemented day. Basically PGF2 alpha is responsible for the onset of
the endogenous production of estradiol from dominant next estrous cycle. Therefore one another manner of
follicle turning Graafian follicle which further enhanced calculation of duration of onset of estrus from the day of
the pre-ovulatory surge, which is responsible for final stage injection PGF2 alpha was also considered for all these
development of the dominant follicle as its one part. protocols. Thus these mean durations of onset of estrus
Likewise in ovsynch protocol 2nd injection of GnRH may from the day of injection of PGF2 alpha, when considered
be considered for the additional release of LH and FSH showed all together different picture from the former.
during the phase which may lead to further enhancement Here this mean duration became the highly significantly
of folliculogenesis than just CIDR-PG based group or longest in the Ovsynch protocol which otherwise from
control group. the end of the protocol was shortest. Buhecha et al (2015)
Table 1. Cohort development of developing follicles
No. Protocol Type No. of developing follicles Total No. of follicles Diameter of dominant
Lt Ovary Rt Ovary follicle(mm)
1. Heatsynch 3.20+0.17 (2-4) 3.33±0.18 (2-4) 6.53±0.27 (4-8) 13.93±0.26b (12-16)
2. Ovsynch 3.33±0.15 (2-4) 3.53±0.13 (3-4) 6.86±0.23 (5-8) 14.2±0.29 b (13-16)
3. CIDR-PG 3.07±0.17 (2-4) 3.23±0.20 (2-4) 6.30±0.20 (5-7) 12.15±0.45a (9-15)
4. C 3.1±0.23 (2-4) 3.4±0.16 (3-4) 6.5±0.26 (5-7) 12±0.53 a (9-14)
Mean values with difference superscripts differ highly significantly (P<0.01).
Table 2. Parameters of induced estrus.
S. No. Protocol Onset of estrus after the Onset of estrous after Estrous duration (h) Diameter of Graafian
end of protocol (h) injection of PG follicle (mm)
1 Heatsynch 37±1.12b (31-46) b
61±1.12 (55-70) 18.46±0.47a (15-22) 19.4±0.93 (14-25)
2 Ovsynch 27.46±0.97a (20-34) 75.46±0.97a (68-82) 18.40±0.58a (14-22) 19.06±0.95 (15-25)
3 CIDR-PG 36.84±1.82b (23-48) 60.84±1.82 b (47-72) 23.53±0.96b (18-31) 18.3±0.67 (15-24)
4. C - - 20.5±0.58a (18-23) 18.2±0.74 (15-21)
Mean values with difference superscripts differ highly significantly (P<0.01)

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Table 3. Insemination done and conception rate
S.N. Protocol Cows showed estrus Cows inseminated
1. Heatsynch 15 (100%) 15 (100%)
2. Ovsynch 15 (100%) 15 (100%)
3. CIDR-PG 13 (100%) 11 (84.61%)
4. C 10 (100%) 9 (90%)
also found longest duration of onset of estrus after
injection of PGF2 alpha in Ovsynch protocol. Zarkouny
(2010) concluded that dairy heifers synchronized with
GnRH and PGF2 alpha should be inseminated when
estrus is observed and heifers that did not exhibit estrus
up to 48 hours after PGF2 alpha injection should be
given a second GnRH injection and fixed time
inseminated 16-20 hours later.
When the mean duration of estrus was considered
it was highest in CIDR protocol then the rest two as well
as the control. Estrogen or GnRH can be said responsible
for hasting of the ovulatory process and thus the duration
of estrus was highly significantly shorter in the heatsynch
and ovsynch protocol. The mean diameter of the Graafian
follicle reveled no significant difference between any of
the groups explains that the protocols did not affected
matrix of the final stage development of Graafian follicle.
Expressing of various signs of estrus like vaginal
edema, amount of mucous discharge, restlessness,
bellowing and tonicity of estrus showed marked
enhancement difference in heatsynch and ovsynch
protocol than CIDR based protocol or control group.
Expression of various signs of estrus like vaginal edema,
amount of mucous discharge, restlessness, bellowing and
tonicity of estrus showed marked enhancement difference
due to extra administration of estradiol in heatsynch
(Colazo and Ambrose, 2013).
Fertility in the synchronized induced estrus
As all the animals taken in each group were cyclic,
thus all the protocols showed 100 induction of estrus
(Table 3) likewise only cyclic dairy cows were taken in
the control group also so the spontaneous occurring of
estrus was also 100 in control group (Mitharwal et al,
2016). Whereas Borakhatariya et al (2017) also obtained
100% estrus induction response during winter season.
Pursley et al (1995) also recommended that
synchronization protocols have major impact on managing
reproduction in lactating dairy cows. Using ovsynch
protocol they recorded 100% cows and 75% heifers
ovulated in the synchronized heat and 50% CR was
recorded in cows. However, they gave 100 µg GnRH
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Cows notinseminated Conception
Nil 9 (60%)
Nil 9 (60%)
2 (15.38%) (Due to purulentdischarge) 6 (54.54%)
1 (10%) (Due to purulent discharge) 6 (66.66%)
instead of 20 µg as in this study as 1st and 3rd shot in the
protocol. In the heatsynch protocol as well as in ovsynch
protocol all the animals showed normal estrus so all the
animals in these groups were inseminated artificially at
the appropriate time of estrus as per the observations of
the signs of estrus. However in CIDR group out of 15, 2
cows dropped CIDR in its course of implantation so those
two cows were excluded from the study. Out of 13, 2
cows showed purulent discharge indicative of infection
in reproductive system and thus were not inseminated
but put on treatment for the condition. In this group
fertility trial was based on the eleven cows inseminations.
Initially the animals taken did not show any sign of
reproductive infection. This means that application of
CIDR led to development of reproductive system infection
probably due to contamination in handling and
application procedures. Likewise, in control one cow
showed purulent discharge indicative of reproductive
infections and thus only nine normal control cows were
inseminated for fertility trials.
Pregnancy diagnosis of 35-60 days in non returned
inseminated cows reveled 60 conception rates (CR) in
the heatsynch and ovsynch protocol, 54.54 in CIDR-
PG based protocol (Table 3). Whereas the control group
recorded 66.66 as highest than any synchronization
protocol. 54 to 60 conception rates of the protocols are
quite acceptable and do not underscore the worth and
significance of heatsynch protocol. Control group
recorded higher conception rate because the estrus cycles
were spontaneous and not artificially interfered whereas
the artificial interference of estrus cycles definitely leads
to some fall in conception rates. Van clef et al (1996)
instead of cyclic cows as in this study, treated dairy heifers
with CIDR for 9 days and PGF2 alpha on day 7. They
recorded 85.1% heifers in estrus 48 to 72 hours after
CIDR removal and 46.4% CR to 1 st insemination. Sa
Filho et al (2010) recorded 81.6% induction of
synchronized heat and 48.7% CR in non suckled cycling
primiparous Bos indicus beef cows when treated with
CIDR for 6 days and then PGF2 alpha. However, unlike
this study they recorded very less 14.6% inseminations
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and 8.5% CR in the control group. That could probably
be due to poor detection of heat in the control group on
feature of beef breed. Rodgers et al (2012) stressed upon
the utility of estrus synchronization protocols and TAI
in commercial cow/calf production in the beef cows
recommended that inspite of cost of protocol, the
economic return in use of such protocols was
advantageous. Geary et al (1998) recorded 59% and 54%
CR in cyclic and non cyclic beef cow. When used
ovsynch protocol and 42% and 38% CR when used
synchro-mate-B in cyclic beef cows, respectively.
Conclusions
In conclusion, among these estrus synchronization
protocols ovsynch protocol was most effective in term of
induction of estrus and conception rate while heatsynch
protocol as most effective for expression of signs of estrus.
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