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Theriogenology 72 (2009) 378385

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Progesterone concentrations, exogenous equine chorionic

gonadotropin, and timing of prostaglandin F2a treatment affect
fertility in postpuberal Nelore heifers
C.C. Dias a, F.S. Wechsler a, M.L. Day b, J.L.M. Vasconcelos a,*
a
Departamento de Producao Animal, Faculdade de Medicina Veterinaria e Zootecnia, FMVZ-UNESP,
CEP 18618-000, Botucatu, SP, Brazil
b
Department of Animal Sciences, The Ohio State University, Columbus, OH 43210, USA
Received 10 September 2008; received in revised form 4 March 2009; accepted 28 March 2009

Abstract
Two experiments were performed to test the hypothesis that elevated progesterone concentrations impair pregnancy rate to timed
artificial insemination (TAI) in postpuberal Nelore heifers. In Experiment 1, postpuberal Nelore heifers (n = 398) received 2 mg
estradiol benzoate (EB) and either a new progesterone-releasing intravaginal device containing 1.9 g of progesterone (CIDR) (first
use) or a CIDR previously used for 9 d (second use) or for 18 d (third use) on Day 0, 12.5 mg prostaglandin F2a (PGF2a) on Day 7,
0.5 mg estradiol cypionate (ECP) and CIDR withdrawal on Day 9, and TAI on Day 11. Largest ovarian follicle diameter was
determined on Day 11. The third-use CIDR treatment increased largest ovarian follicle diameter and pregnancy rate. Conception to
TAI was reduced in heifers with smaller follicles in the first- and second-use CIDR treatments, but not in the third-use CIDR
treatment. In Experiment 2, postpuberal Nelore heifers received the synchronization treatment described in Experiment 1 or
received 12.5 mg PGF2a on Day 9 rather than Day 7. In addition, 50% of heifers received 300 IU equine chorionic gonadotropin
(eCG) on Day 9. Heifers were either TAI (Experiment 2a; n = 199) or AI after detection of estrus (Experiment 2b; n = 125 of 202).
In Experiment 2a, treatment with eCG increased pregnancy rate to TAI in heifers that received PGF2a on Day 9 but not on Day 7 and
in heifers that received a first-use CIDR but not in heifers that received a third-use CIDR. Treatments did not influence reproductive
performance in Experiment 2b. In summary, pregnancy rate to TAI in postpuberal Nelore heifers was optimized when lower
concentrations of exogenous progesterone were administered, and eCG treatment was beneficial in heifers expected to have greater
progesterone concentrations.
# 2009 Elsevier Inc. All rights reserved.

Keywords: Cattle; Pregnancy; Proestrus; Progesterone; Timed artificial insemination

1. Introduction
Artificial insemination (AI) is the most effective
technique available to introduce new genetics into a
* Corresponding author. Tel.: +55 14 38117185;
fax: +55 14 38117180.
E-mail address: vasconcelos@fca.unesp.br (J.L.M. Vasconcelos).
beef cattle herd, and timed AI (TAI) protocols have been
developed to overcome challenges associated with
estrus detection failure. There is a need for development
of TAI protocols for Bos indicus breeds because of a
shorter duration of estrous behavior compared with that
of Bos taurus breeds [1].
Low TAI pregnancy rate was reported in Nelore
heifers in studies in which the estrous synchronization
protocol was initiated with estradiol administration and

0093-691X/$ see front matter # 2009 Elsevier Inc. All rights reserved.
doi:10.1016/j.theriogenology.2009.03.006
 C.C. Dias et al. / Theriogenology 72 (2009) 378385
insertion of a progesterone-releasing intravaginal
device containing 1.9 g of progesterone (CIDR) [2,3].
Carvalho et al. [4] compared the interval from CIDR
insertion and estradiol treatment to follicular wave
emergence in Bos indicus (3.1 d), Bos indicus  Bos
taurus (3.3 d), and Bos taurus (3.2 d) heifers. Although
the time of follicle emergence was similar among
groups, Bos indicus heifers had the smallest dominant
follicles at the end of the CIDR treatment. A lower
growth rate of dominant follicles in the Bos indicus
heifers appeared to be associated with greater circulat-
ing progesterone concentrations during CIDR treat-
ment. These results suggested that high concentrations
of progesterone during estrous synchronization protocol
may be detrimental to follicular development in Bos
indicus cattle. Because luteinizing hormone (LH)
secretion and pulse frequency are negatively associated
with progesterone concentration [5], perhaps endogen-
ous gonadotropic support for follicular development
may be compromised in progesterone-treated Bos
indicus heifers, as described by Carvalho et al. [4].
An alternative for increasing gonadotropic support of
dominant follicles before TAI is a longer interval from
luteal regression to the LH surge and TAI (i.e.,
proestrus) than that of traditional synchronization
protocols. Pregnancy rate to AI has been demonstrated to
be greater in cows that had a long proestrus [610]. Use of
equine chorionic gonadotropin (eCG) is a second
alternative to increase follicle development; eCG treat-
ment increased conception rate to TAI when given at the
end of a CIDR treatment in anestrous cows [11]. A third
alternative may be decreasing progesterone concentra-
tions during the synchronization protocol. The goal of the
current experiments was to determine whether treatments
that should increase gonadotropic support of follicular
development will increase fertility in Bos indicus heifers.
The overall hypothesis for these experiments was
that low pregnancy rates to TAI previously reported in
Nelore heifers were due to inhibition of LH secretion
and in turn development of the ovulatory follicle caused
by elevated systemic progesterone concentrations. The
objective of the first experiment was to compare the
effect of various concentrations of progesterone during
a synchronization protocol on pregnancy rate to TAI by
using CIDRs that were expected to deliver different
concentrations of progesterone. In the second experi-
ment, progesterone concentrations were altered by
either changing the timing of luteal regression during a
synchronization protocol and/or using CIDRs that
would deliver different doses of progesterone. In
addition, treatment with eCG was used to evaluate
the effect of an exogenous gonadotropic support. The
379
influence of these factors on follicular development was
assessed in both TAI and AI after estrus detection.
2. Materials and methods
2.1. Experimental design, location, and heifers
2.1.1. Experiment 1
This trial was conducted between November and
December 2005 in Mato Grosso do Sul, Brazil. Nelore
heifers (n = 525), 24 to 26 mo of age, with body
condition scores (BCSs) between 2.75 and 3.25 (1 to 5
scale [12]) were used. Heifers were maintained on two
pastures (Brachiaria humidcola) with mineral salt ad
libitum. Their ovaries were evaluated by ultrasono-
graphy for presence of a corpus luteum (CL; Aloka SSD
500 V, equipped with a 7.5-MHz linear array intrarectal
transducer; Aloka, Tokyo, Japan) 7 d before the
initiation of the protocol. Heifers with luteal tissue in
their ovaries (n = 398) were randomly assigned to
receive a new progesterone-releasing intravaginal
device containing 1.9 g progesterone (CIDR; Pfizer
Animal Health, Sao Paulo, SP, Brazil; new CIDR; first
use), a CIDR that had been used previously in a 9-d
synchronization protocol (second use), or a CIDR that
had been used in two previous 9-d synchronization
protocols (third use). All heifers were treated with 2 mg
estradiol benzoate (EB) im (Estrogin; Farmavet, Sao
Paulo, SP, Brazil) at CIDR insertion (Day 0 of the
experiment). Blood samples for progesterone analyses
were collected on Days 0 and 7, and prostaglandin F2a
(PGF2a) was administered (12.5 mg, im; Lutalyse;
Pfizer Animal Health) on Day 7. On Day 9, CIDRs were
withdrawn, and heifers received estradiol cypionate
(0.5 mg, im; estradiol cypionate (ECP); Pfizer Animal
Health). Heifers were bred by TAI on Day 11 (48 h after
CIDR withdrawal) by two technicians using frozen-
thawed semen from a single sire. Transrectal ultra-
sonography examinations were performed on Day 11 to
assess diameter of largest ovarian follicle at TAI
(defined as the average between horizontal and vertical
diameters). Heifers in which the largest ovarian follicle
was <6 mm were recorded as a missing observation for
this variable. Ultrasonography was also performed on
Day 13 to evaluate ovulation (absence of the largest
ovarian follicle previously measured at TAI) and on Day
41 to detect pregnancy. Reproductive variables that
were determined included ovulation, synchronization,
conception, and pregnancy rates. Ovulation rate was
defined as the proportion of all heifers that ovulated
between Days 11 and 13, including heifers that were
considered to have prematurely ovulated (presence of
380 C.C. Dias et al. / Theriogenology 72 (2009) 378385
multiple small follicles that were <6 mm in diameter at
TAI). Synchronized ovulation rate was defined as the
proportion of all heifers that ovulated between Days 11
and 13. Conception rate was calculated as the
proportion of synchronized heifers that were pregnant
on Day 41, and pregnancy rate was calculated as the
proportion of all heifers that were pregnant at that time.
2.1.2. Experiment 2
This experiment was conducted between November
2006 and January 2007 in Mato Grosso do Sul, Brazil.
Nelore heifers (n = 650), 24 to 26 mo of age, with BCSs
between 2.75 and 3.25 were observed for estrus for 21 d
before initiation of the synchronization program.
Heifers detected in estrus (n = 401) were used in the
experiment, as they were considered as to be
reproductively mature. Heifers were maintained on
two pastures (Brachiaria humidcola) with mineral salt
provided ad libitum. On Day 0, all heifers received 2 mg
EB im and were randomly assigned to receive either a
first-use, second-use, or third-use CIDR as described in
Experiment 1, PGF2a (12.5 mg, im) given on Day 7 or
Day 9, and either no eCG or 300 IU eCG im (Folligon;
Intervet, Sao Paulo, SP, Brazil) on Day 9.
Heifers from the two pastures had their CIDRs
withdrawn on Day 9 and were randomly assigned either
to receive ECP (0.5 mg, im) at this time, followed by
TAI on Day 11 (Experiment 2a; n = 199), or to be
artificially inseminated 12 h after detection of estrus
between Days 9 and 15 (Experiment 2b; n = 202). All
heifers were artificially inseminated by two technicians
using frozen-thawed semen from a single sire.
Pregnancy diagnoses were performed by transrectal
ultrasonography 30 d after TAI.
2.2. Blood samples and progesterone RIA analyses
2.2.1. Experiment 1
Blood samples for progesterone analyses were
collected from a coccygeal vessel into Vacutainer tubes
(Becton Dickinson Co., Franklin Lakes, NJ, USA).
Blood was allowed to clot at 4 8C for 24 h and
centrifuged at 1500  g for 15 min at room tempera-
ture. Serum was removed and frozen at 20 8C until
assays were performed. Serum progesterone concentra-
tions were determined using a solid-phase radio-
immunoassay kit containing antibody-coated tubes
and 125I-labeled progesterone (Coat-a-count; Diagnos-
tic Products Corporation, Los Angeles, CA, USA) that
had been previously validated in our laboratory [13].
The intra-assay coefficients of variation were 6.7% and
8.5% for assays 1 (Day 0) and 2 (Day 7), respectively,
and the interassay coefficient of variation was 5.4%.
Sensitivity was 0.01 ng/mL.
2.3. Statistical analyses
2.3.1. Experiment 1
Continuous dependent variables (i.e., progesterone
concentration on Day 7 and diameter of the largest
ovarian follicle on Day 11) were analyzed by least-
squares (PROC GLM; SAS Institute Inc., Cary, NC,
USA [14]) using a model that included treatment (first-
use, second-use, and third-use CIDRs), progesterone
concentration on Day 0, and the interaction. Progester-
one concentrations (Days 0 and 7) and largest ovarian
follicle diameter were tested for normality (Kolmo-
gorov-Smirnov test) and homogeneity of variances
(Brown-Forsythe test). Progesterone concentrations
were submitted to a log transformation due to
heterogeneity of variance. Specific mean comparisons
were made using the Tukey-Kramer test. A secondary
analysis was performed to understand the factors that
contribute to variation in largest ovarian follicle
diameter. To accomplish this, a model that included
treatment, progesterone concentration on Day 0,
progesterone concentration on Day 7, and the relevant
interactions, with BCS as a covariate, was used. The
GLM procedure of SAS was used for this analysis. The
model was reduced by sequentially removing non-
significant (P > 0.10) terms (backwards stepwise
regression), with the final model including treatment
and progesterone concentration on Day 0.
Dependent binomial variables (ovulation, synchro-
nization, conception, and pregnancy rate) were ana-
lyzed by logistic regression (PROC LOGISTIC; SAS
Institute Inc., Cary, NC, USA [14]) using a model that
included treatment (first-use, second-use, and third-use
CIDRs), progesterone concentration on Day 0, the
interaction, and BCS as a covariate. For conception and
pregnancy rate, AI technician and the appropriate
interactions were also included in the model. Specific
mean comparisons were made using the Bonferroni test.
Secondary analyses were performed to understand the
relationships of treatment, progesterone concentrations
on Days 0 and 7, and largest ovarian follicle diameter
with dependent binomial variables. For ovulation rate,
the model included treatment, progesterone concentra-
tion on Day 0, progesterone concentration on Day 7, and
the relevant interactions (with BCS as a covariate). For
pregnancy rate, AI technician and interactions were
added to the model, and largest ovarian follicle diameter
was removed. For synchronization and conception
rates, largest ovarian follicle diameter and interactions
 C.C. Dias et al. / Theriogenology 72 (2009) 378385 381

were also added. Logistic regression was used for these
analyses, with the model reduced by sequentially
removing non-significant (P > 0.10) terms (backwards
stepwise regression).
In a final analysis, heifers were categorized based on
largest ovarian follicle diameter as previously described
by Gimenes et al. [15]. Categories were Class 1
(follicles smaller than 8.5 mm), Class 2 (between 8.5
and 10 mm), and Class 3 (larger than 10 mm).
Synchronized ovulation rate and conception rate were
analyzed using the Logistic procedure of SAS, with
treatment, largest ovarian follicle class, and the
interaction included in the model. Specific mean
comparisons were made using the least square means
(LSM) procedure.
2.3.2. Experiment 2
Dependent binomial variables, pregnancy rate after
TAI (Experiment 2a), percentage of heifers detected in
estrus, and conception rate of these heifers (Experiment
2b) were analyzed by logistic regression (PROC
LOGISTIC; SAS Institute Inc.) using a model that
included CIDR uses (first, second, or third), day of
PGF2a, eCG, AI technician, and the appropriate
interactions. The model was reduced by sequentially
removing non-significant (P > 0.10) terms (backwards
stepwise regression), with the final model for Experi-
ment 2a including CIDR uses, eCG, day of PGF2a,
CIDR uses by eCG, and day of PGF2a by eCG. For
Experiment 2b, the reduced model included CIDR uses,
day of PGF2a, and eCG. Specific mean comparisons
were made using the Bonferroni test.
For both experiments, probability values of a  0.05
were considered significant, whereas probability values
of 0.05 < a  0.1 were considered as tendencies.
3. Results
3.1. Experiment 1
3.1.1. Progesterone concentrations and largest
ovarian follicle diameter
On Day 0, concentration of progesterone did not
differ among treatments (2.8  2.4 ng/mL). On Day 7,
progesterone concentrations were lower (P < 0.05) in
the third-use CIDR group than in the first-use CIDR
group; with the second-use CIDR group intermediate
(P > 0.1, Table 1). Concentrations of progesterone on
Day 7 were not influenced by concentrations of
progesterone on Day 0. Diameter of the largest ovarian
follicle on Day 11 was greater (P < 0.05) for the third-
use CIDR treatment than for the first-use CIDR
treatment and tended (P < 0.10) to be greater than that
for the second-use CIDR treatment (Table 1). Addi-
tionally, serum progesterone concentrations on Day 0
were associated with largest ovarian follicle diameter
(Fig. 1). In the secondary analysis, progesterone
concentrations on Day 7 did not influence largest
ovarian follicle diameter on Day 11.
3.1.2. Reproductive parameters
Neither ovulation (73.6%) nor synchronized ovula-
tion rate (68.7%) were influenced by treatment or
progesterone concentration on Day 0 (Table 1). How-

Table 1
Serum progesterone concentration on Day 7, diameter of largest ovarian follicle at TAI, ovulation, synchronized rates of ovulation, conception
(synchronized heifers), and pregnancy related to CIDR treatments (Experiment 1).
CIDR Serum P4 on Follicle diameter Ovulation rate,6 Synchronized Conception rate,8 Pregnancy rate,9
Day 7 4 at TAI,5 mm (n) % (n) ovulation rate,7 % (n) % (n) % (n)
First use1 3.0  1.9 A 10.1  2.0A (121) 70.5A (103/146) 64.5A (78/121) 37.2a (29/78) 20.5A (30/146)
Second use2 2.3  1.6A,B 10.6  2.3A,B, a (109) 72.7A (93/128) 67.9A (74/109) 37.8a (28/74) 22.7A (29/128)
Third use 3 2.0  1.2 B 11.0  2.1B, b (106) 78.2A (97/124) 74.5A (79/106) 53.2b (42/79) 35.5B (44/124)
Total 2.5  1.6 10.5  2.2 (336) 73.6 (293/398) 68.7 (231/336) 42.9 (99/231) 25.9 (103/398)
A,B
Values with different superscripts within a column are different (P < 0.05).
a,b
Values with different superscripts within a column tended to differ (P < 0.1).
1
CIDR devices with no previous use.
2
CIDR devices with 9 d of previous use.
3
CIDR devices with 18 d of previous use.
4
Raw means  SD for serum progesterone concentrations on Day 7 of the protocol.
5
Raw means  SD for diameter of largest ovarian follicle at TAI.
6
Percentage of all heifers that ovulated by 48 h after TAI, including heifers that ovulated between CIDR removal and TAI, and within 48 h after TAI,
reported as raw mean.
7
Percentage of heifers that ovulated within 48 h after TAI, reported as raw mean.
8
Percentage of synchronized ovulated heifers that became pregnant, reported as raw mean.
9
Percentage of all heifers pregnant after treatment with synchronization protocol, reported as raw mean.
382 C.C. Dias et al. / Theriogenology 72 (2009) 378385
Fig. 1. Effect (P < 0.05) of serum progesterone concentrations at the
initiation of a synchronization protocol on diameter of largest ovarian
follicle at TAI in postpuberal Nelore heifers. The pooled SEM value
for diameter of the largest ovarian follicle at TAI was 0.22 mm.
Fig. 2. Effect (P < 0.05) of serum progesterone concentration at the
initiation of a synchronization protocol on the probability of concep-
tion to TAI in postpuberal Nelore heifers.
ever, conception rate tended (P < 0.10) to be greater for
the third-use treatment than those for second-use and
first-use CIDR treatments. Heifers with greater pro-
gesterone concentrations on Day 0 had greater
(P < 0.05) conception rate (Fig. 2). Pregnancy rate
was also greater (P < 0.05) for the third-use treatment
than those for the second-use and first-use CIDR
treatments and in heifers with greater concentrations of
progesterone on Day 0 (Fig. 3). Neither BCS nor AI
technician were a significant source of variation. In the
secondary analyses, progesterone on Day 7 did not
affect ovulation, synchronization, conception, or preg-
nancy rate. Diameter of the largest ovarian follicle on
Fig. 3. Effect (P < 0.05) of serum progesterone concentration at the
initiation of a synchronization protocol on the probability of preg-
nancy to TAI in postpuberal Nelore heifers.
Fig. 4. Effect of largest ovarian follicle diameter at TAI (P < 0.05) on
the probability of synchronized ovulation in postpuberal Nelore
heifers.
Fig. 5. Effect of diameter of largest ovarian follicle at TAI (P < 0.05)
on the probability of conception to TAI in postpuberal Nelore heifers.
Day 11 influenced (P < 0.05) rates of synchronization
(Fig. 4) and conception (Fig. 5).
When heifers were divided into largest ovarian
follicle classifications (Class 1, 2, and 3), synchronized
ovulation rate increased (P < 0.05) with each largest
ovarian follicle class. Synchronized ovulation rate was
greatest for heifers with Class 3 largest ovarian follicle
(92.2%; 141 of 153), intermediate for heifers with Class
2 largest ovarian follicle (62.5%; 70 of 112), and least
for heifers with Class 1 largest ovarian follicle (28.2%;
20 of 71). A treatment by largest ovarian follicle class
interaction was detected (P < 0.05) for conception rate
(Table 2). In the first-use and second-use CIDR
treatments, conception rate was greater (P < 0.05) in
heifers with Class 3 largest ovarian follicle than those
for Class 1 or 2. In the third-use CIDR treatment, largest
ovarian follicle class did not affect conception rate.
3.2. Experiment 2
3.2.1. Experiment 2a
Interactions of day of PGF2a and eCG treatment
(P < 0.05) and CIDR uses and eCG treatments
(P < 0.05) were detected for TAI pregnancy rate
(Table 3). The eCG treatment enhanced (P < 0.05)
pregnancy rate in heifers that received PGF2a on Day 9
 C.C. Dias et al. / Theriogenology 72 (2009) 378385 383

Table 2
Conception rate (synchronized heifers) related to the diameter of largest ovarian follicle at TAI classes within each CIDR treatment (Experiment 1).
CIDR Conception rate,4 % (n) P Value
5 6 7
Class 1 Class 2 Class 3
1 a a
First use 9.1 (1/11) 24.0 (6/25) 52.4 (22/42)b <0.05
Second use2 0 (0/4)a 21.7 (5/23)a 48.9 (23/47)b <0.05
Third use 3 40.0 (2/5)a 54.5 (12/22)a 53.8 (28/52)a >0.10
Class = classification of largest ovarian follicle at TAI.
a,b
Values with different superscripts within a row are different (P < 0.05).
1
CIDR devices with no previous use.
2
CIDR devices with 9 d of previous use.
3
CIDR devices with 18 d of previous use.
4
Percentage of heifers that ovulated within 48 h after TAI that became pregnant, reported as raw means.
5
Heifers with follicles <8.5 mm at TAI.
6
Heifers with follicles between 8.5 and 10.0 mm at TAI.
7
Heifers with follicles >10.0 mm at TAI.

but not in those that received PGF2a on Day 7. The eCG (40.1%; 81 of 202) were not affected by the CIDR,
treatment also increased pregnancy rate in heifers PGF2a, or eCG treatments.
receiving the first-use CIDR (P < 0.05); tended to
increase pregnancy rate with a second-use CIDR 4. Discussion
(P < 0.1); but had no influence on pregnancy rate in
the third-use CIDR treatment. The TAI pregnancy rate was increased when either
progesterone was decreased during the synchronization
3.2.2. Experiment 2b protocol or eCG was administered. None of the
Overall rates of estrus (61.9%; 125 of 202), manipulations negatively influenced fertility if the
conception (64.8%; 81 of 125), and pregnancy heifers were permitted to exhibit estrus and AI was
performed based on detection of this physiologic end
Table 3 point.
Pregnancy rate in postpuberal Nelore heifers at TAI protocol, with In the first experiment, pregnancy rate to TAI was
PGF2a on Days 7 or 9, with or without an eCG treatment, and with greater in heifers treated with the third-use CIDR than
CIDR treatments of varying previous uses (Experiment 2a). with other CIDR treatments. In addition, concentrations
Treatments Pregnancy at TAI,6 % (n) of progesterone were different on Day 7. These
Without eCG7 With eCG8
differences were similar to those of previous findings
in our laboratory [16]. A statistical relationship between
PGF2a progesterone concentration on Day 7 and pregnancy
Day 7 1 33.3 (16/48)A 40.4 (21/52)A
Day 9 2 26.4 (14/53)A 50.0 (23/46)B
rate was not evident, suggesting that the influence of the
CIDR CIDR on progesterone at this time was not a primary
First use3 21.9 (7/32)A 50.0 (18/36)B determinant of fertility. Because progesterone concen-
Second use4 24.3 (9/37)a 42.4 (14/33)b trations after Day 7 would be largely determined by type
Third use 5 43.7 (14/32)A 41.4 (12/29)A of CIDR (due to luteal regression), it is logical to
A,B
Values for the same variable with different superscripts within a presume that the primary effect of the type of CIDR
row are different (P < 0.05). would be emphasized from Days 7 to 9. That
a,b
Values for the same variable with different superscripts within a row
gonadotropic support of the follicle may have been
tended to differ (P < 0.1).
1
PGF2a (12.5 mg, im) treatment on Day 7 of synchronization protocol. enhanced in the third-use CIDR treatment was
2
PGF2a (12.5 mg, im) treatment on Day 9 of synchronization protocol. reinforced by the increase in follicular diameter at
3
CIDR devices with no previous use. TAI that was detected in this treatment. Because the
4
CIDR devices with 9 d of previous use. interval from insertion of a CIDR and estradiol
5
CIDR devices with 18 d of previous use. treatment to follicular wave emergence was 3.1 d in
6
Percentage of all heifers that became pregnant after treatment with
synchronization protocol, reported as raw means. Bos indicus [4], and deviation occurred 1.6 d after wave
7
No eCG treatment on Day 9 of synchronization protocol. emergence, within diameter of the largest ovarian
8
eCG treatment (300 IU, im) on Day 9 of synchronization protocol. follicle of 5.9 mm at the beginning of deviation [17] or
384 C.C. Dias et al. / Theriogenology 72 (2009) 378385
6.2 mm [15], and this is when dominant follicles
acquire LH receptors [18], the ability of increased
gonadotropic support (due to decreased progesterone)
to enhance follicular maturation may have be opti-
mized.
Larger follicles at the time of AI and conception and
pregnancy rates to TAI were associated with high
progesterone concentrations on Day 0. In the current
study, the Day 0 progesterone concentration sample was
collected before the insertion of CIDR devices, and it is
related to the endogenous progesterone. We inferred
that heifers with greater progesterone concentrations at
the outset of the protocol were in later stages of the
estrous cycle, and thus, spontaneous luteal regression
occurred during the protocol and contributed to this
response.
Follicular diameter at TAI was related to synchro-
nization and conception rates across treatments.
Heifers with larger follicles were more likely to
ovulate at the appropriate time and conceive. Relation-
ships between largest ovarian follicle diameter and
synchronized ovulation heifers and conception rate
were revealed when heifers were categorized into
largest ovarian follicle diameter classes. Synchronized
ovulation rate increased as largest ovarian follicle
diameter increased, regardless of CIDR treatment.
Within the third-use CIDR treatment, largest ovarian
follicle classification did not influence conception rate,
whereas with first-use and second-use CIDRs, con-
ception was reduced in heifers with smaller ovarian
follicles at TAI. We inferred that if progesterone
concentrations during the later stages of the estrous
synchronization protocol are reduced, the range of
largest ovarian follicle diameters that results in
acceptable fertility is increased. We speculate that
when progesterone concentrations are reduced, gona-
dotropic stimulus is enhanced, most likely through LH
concentrations, improving follicle maturity and
increasing fertility.
In Experiment 2a, the effect of eCG was dependent
on timing of PGF2a administration (Days 7 or 9) and
CIDR type. The reduced fertility in heifers with high
progesterone concentrations (i.e., PGF2a on Day 9 or
first-use and second-use CIDRs) could be overcome by
administration of eCG at CIDR withdrawal.
Strategies to reduce circulating progesterone con-
centration during later stages of a TAI synchronization
protocol and/or increase gonadotropic stimulus
improved reproductive performance in postpuberal
Nelore heifers, and conversely, those that resulted in
greater progesterone concentrations in the later stages
of the TAI protocol negatively affected reproductive
performance. Timing of this adjustment in progesterone
concentrations may be important, as a recent report in
Nelore heifers demonstrated that treatment with PGF2a
concurrent with CIDR insertion did not improve
pregnancy rate using TAI [2]. The range of ovarian
follicle diameters that resulted in acceptable fertility
appeared to be extended in heifers with the lesser
progesterone concentrations. We speculate that LH
secretion in Nelore heifers is highly sensitive to
progesterone negative feedback and that use of low
concentrations of progesterone increases gonadotropic
support for follicular maturation. It also suggests that
synchronization protocols in postpuberal Nelore heifers
should be designed to reduce concentrations of
progesterone during maturation of the ovulatory
follicle.
In conclusion, postpuberal Nelore heifers need a
period of reduced progesterone concentrations, as
physiologically occurs in their estrous cycle, during
the later stages of a synchronization protocol. Further-
more, gonadotropin treatment may be useful to
optimize pregnancy rate to TAI in Nelore heifers
treated with progesterone devices that will increase
circulating progesterone to more than high physiologic
concentrations.
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