Theriogenology 86 (2016) 1268–1274

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Theriogenology
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Luteolysis in Bos indicus cows on Days 5 and 7 of estrous

cycle with varying doses of PGF2a
Marcos Vinicius C. Ferraz Junior a, Alexandre V. Pires a, b, *,
Marcos Vinicius Biehl b, Marcelo Henrique Santos a, Daniel M. Polizel a,
Delci D. Nepomuceno b, Roberto Sartori b, João Bosco Barreto Filho c,
José Renato S. Gonçalves d, Michael L. Day e
a
Department of Nutrition and Animal Production, Faculty of Veterinary and Animal Science, University of São Paulo, Pirassununga,
São Paulo, Brazil
b
Department of Animal Science, College of Agriculture “Luiz de Queiroz” (ESALQ), University of São Paulo, Piracicaba,
São Paulo, Brazil
c
Department of Veterinary, Federal University of Lavras, Lavras, Minas Gerais, Brazil
d
Experimental Station Hildegard Georgina Von Pritzelwitz, Fazenda Figueira, Londrina, Paraná, Brazil
e
Department of Animal Sciences, The Ohio State University, Columbus, Ohio, USA

a r t i c l e i n f o a b s t r a c t

Article history: The aim of this study was to evaluate luteolysis using three doses of PGF2a on Day 5 or Day
Received 7 July 2015 7 of the estrous cycle in nonlactating Nellore (Bos indicus) cows. Cows (n ¼ 323) were
Received in revised form 14 April 2016 assigned within date of estrus (Day 0 of estrous cycle) to receive 12.5, 25.0, or 50.0 mg of
Accepted 18 April 2016
PGF2a on either Day 5 or Day 7 of the estrous cycle in a 3  2 factorial arrangement. Blood
samples for progesterone (P4) concentrations were collected at 0, 24, 48, and 72 hours after
Keywords:
PGF2a to assess luteolysis (L). Luteolysis was defined on the basis of P4 concentrations at
Luteolysis
72 hours using either less than 0.5 ng/mL (L0.5) or less than 1.0 ng/mL (L1.0) as the cut off.
PGF2a dose
Fixed time artificial insemination Luteolysis was considered “partial” when P4 concentration declined within 24 hours after
PGF2a but failed to decline further or, in some cases, increased. Incidence of luteolysis was
less (P < 0.01) on Day 5 than Day 7 of the estrous cycle (17.3 vs. 47.6% and 30.4 vs. 77.2%;
for L0.5 and L1.0, respectively). Dose of PGF2a increased (P < 0.01) L1.0 (12.5 mg ¼ 38.9%;
25.0 mg ¼ 52.3%; and 50.0 mg ¼ 70.4%). Incidence of partial luteolysis for cows on Day 5
(57.1%) was greater (P < 0.01) than that on Day 7 (19.1%) of the estrous cycle and was more
prevalent (P < 0.01) with lower doses of PGF2a (12.5 mg ¼ 49.1%; 25.0 mg ¼ 37.4%; and
50.0 mg ¼ 27.8%). In conclusion, both days of the estrous cycle and doses of PGF2a influ-
enced the incidence of complete and partial luteolysis in Nellore cows and should be an
important consideration when devising estrus synchronization programs in this species.
Ó 2016 Elsevier Inc. All rights reserved.

1. Introduction receptors in the early CL [2,3], luteolysis from exogenous

Prostaglandin F2a is a hormone that triggers luteolysis in
cows, inducing the end of luteal phase [1]. However, despite
the presence of similar concentrations of high-affinity PGF2a
* Corresponding author. Tel.: 55 (19) 3429-4247; fax: 55 (19) 3429-
4215.
E-mail address: pires.1@usp.br (A.V. Pires).
PGF2a does not usually occur until 5 to 6 days after estrus
[4–9]. An important aspect of synchronization programs that
use GnRH to synchronize emergence of a new follicular wave
through induction of ovulation of the dominant follicle is to
ensure lysis of the CL 5 to 7 days later in the Ovsynch- and/or
CO-Synch–based protocols [10,11]. The mechanisms of
action of PGF2a in newly formed CL are not completely
understood [6,7,9].

0093-691X/$ – see front matter Ó 2016 Elsevier Inc. All rights reserved.
http://dx.doi.org/10.1016/j.theriogenology.2016.04.067
 M.V.C. Ferraz Junior et al. / Theriogenology 86 (2016) 1268–1274
Studies reported that luteolysis occurred in more than
90% in cows that received a single (25 mg) dose of PGF2a
after Day 8 of the estrous cycle [11]. In contrast, a single
(25 mg) dose of PGF2a in the first 4 days of the cycle did not
induce luteolysis [6,7]. However, the incidence of luteolysis
induced by a double dose of PGF2a (50 mg) around Day 5 of
the estrous cycle is not well established [4,5,7]. In addition,
studies have demonstrated that a transient decrease in
circulating progesterone (P4) concentration in cows does not
always indicate complete luteolysis when PGF2a is admin-
istered around Day 5 of the estrous cycle [5,6]. This response
is referred to as partial luteolysis and is characterized as a
decrease in P4 concentration during the first 24 hours after
PGF2a, but after that, no further decline or, in some cases, a
rebound in systemic concentrations occur [5,6,12].
Ensuring complete luteolysis and total decrease in
circulating P4 at the time of artificial insemination (AI) is
essential for optimizing results in fixed timed AI (FTAI)
protocols [13]. To induce complete luteolysis in recently
formed CL, several strategies have been studied, including
the use of a greater dose of PGF2a and/or multiple doses of
PGF2a given at varying intervals [6,7].
In recent years, incidence of luteolysis during the first
days of the estrous cycle has been investigated in multiple
experiments [6,7]. All these studies have used Bos taurus
cows as the animal model. The objective of the present
study was to investigate the efficacy of differing PGF2a
doses to induce luteolysis on Day 5 or Day 7 of the estrous
cycle in Nellore cows.
2. Materials and methods
The experiment was performed in the Experimental
Station Agrozootécnica Hildegard Georgina Von Pritzel-
wiltz (23 340 2500 S; 50 580 1700 W), in Londrina, Paraná,
Brazil, which belongs to the Fundação de Estudos Agrários
Luiz de Queiroz.
Nonlactating, cyclic Nellore cows (n ¼ 323) with no
uterine or ovarian abnormalities (based on a transrectal
scan of the reproductive tract by ultrasonography) were
used. Groups of approximately 80 cows at random stages of
their estrous cycle received 25.0 mg of PGF2a (dinoprost
tromethamine–Lutalyse, Zoetis, São Paulo, SP, Brazil).
Estrus detection was performed twice daily for 5 days;
aided by vasectomized bulls fitted with chin-ball markers.
Cows detected in estrus were assigned, on the day of
estrus, to receive one of three doses of PGF2a on either Day 5
or Day 7 of the ensuing estrous cycle. Doses were 12.5, 25.0,
or 50.0 mg of PGF2a given on Day 5 or Day 7 of the estrous
cycle. The 3  2 factorial arrangement resulted in six treat-
ments: Day 5/12.5PGF2a (n ¼ 54), Day 5/25.0PGF2a (n ¼ 54),
Day 5/50.0PGF2a (n ¼ 53), Day 7/12.5PGF2a (n ¼ 54), Day
7/25.0PGF2a (n ¼ 53), and Day 7/50.0PGF2a (n ¼ 55).
Ovulation was confirmed by detection of CL by ultra-
sonography (US) on Day 5 or Day 7, and CL diameter was
recorded when PGF2a was given (0 hours) and at 96 hours
to assess morphologic luteolysis. Estrus detection was
performed twice daily, aided by the use of vasectomized
bulls fitted with chin-ball markers for a period of 120 hours.
Blood samples (10 mL) were collected via a coccygeal
vessel at 0, 24, 48, and 72 hours after PGF2a administration.
1269
Blood samples were centrifuged for 15 minutes at 1800  g.
Serum was frozen at 20  C for further analysis. Proges-
terone concentrations were measured by chemilumines-
cent assay using commercial IMMULITE 1000 kits (Siemens
Healthcare Diagnostics, Deerfield, IL, USA). All the analyses
were performed in the Nutrition and Animal Reproduction
Laboratory–LZT/ESALQ/USP. The coefficients of variation
were 2.7% and 3.6% for low and high adjuster, respectively.
Luteolysis was determined to have occurred when P4
concentrations were less than 1.0 ng/mL (L1.0) or less than
0.5 ng/mL (L0.5) 72 hours after PGF2a. When P4 concen-
trations declined during the first 24 hours after treatment
but did not decline further, or in some cases increased, this
was classified as partial luteolysis. If P4 concentration did
not decrease during the 24 hours after PGF2a, this was
classified as failure of luteolysis.
A randomized experimental design in a 3  2 factorial
arrangement (day of estrous cycle  PGF2a dose) was used.
The variables were analyzed after running normality (Sha-
piro–Wilk) and homogeneity of variance (Welch) tests.
Variables with non-normal distribution were submitted to
either logarithmic (log10 [x þ 1]) or quadratic (SQRT [x þ 1/
2]) transformation before analysis. Variables such as CL
diameter at 0 and 96 hours after PGF2a administrated and
the interval to estrus were analyzed using the MIXED pro-
cedure. The same procedure with repeated measures over
time analyses was used to evaluate P4 concentrations at 0,
24, 48, and 72 hours after PGF2a in each day of the estrous
cycle (Day 5 and Day 7), separately, according to PGF2a dose.
Three analyses of P4 concentrations in each day of the
estrous cycle (Day 5 and Day 7) were performed. The first
analysis considered all cows, the second, only cows with
complete luteolysis, and the third considering only cows
with partial luteolysis. The GLIMMIX procedure was used for
variables with binomial distribution, such as L0.5, L1.0, par-
tial luteolysis rate, no luteolysis, and incidence of estrus.
The effect of relative drop in circulating P4 concentration
(100  [P4 concentration at 24 hours after PGF2a  100]/P4
concentration at 0 hour at PGF2a) on estrous behavior was
analyzed by logistic regression (LIFETEST procedure)
comparing PGF2a doses or days of the estrous cycle when
PGF2a treatments were employed. The oddsratio statement
was used to compare difference among doses of PGF2a.
Logistic regression curves were done using the coefficient
generated by Interactive Data Analysis of SAS
(logit ¼ intercept þ slop  [drop in P4]). Probability curve
was made by Y ¼ (EXP [logit]/1 þ EXP [logit])  100. Sig-
nificant differences were considered when P < 0.05. All
procedures used were from SAS 9.3 statistical software.
3. Results
There was no interaction between PGF2a doses and day
of the estrous cycle in any of the analyzed variables. Dose of
PGF2a did not alter the time of estrus after PGF2a
(70.2  1.76 hours), and 50.4% of the cows that exhibited
estrus showed estrus within 72 hours after PGF2a (Fig. 1).
However, the mean time of estrus was later (P < 0.01) in
cows treated on Day 5 than those on Day 7 (76.0  3.12 vs.
66.0  2.29 hours, respectively). Cows treated with PGF2a
on Day 7 were more likely to exhibit estrus (P < 0.01) than
1270 M.V.C. Ferraz Junior et al. / Theriogenology 86 (2016) 1268–1274

Fig. 2. Probability of estrus according to relative drop (%) in progesterone (P4)

Fig. 1. Estrus distribution according with PGF2a doses (12.5, 25.0, or 50.0 mg
of dinoprost tromethamine) administered on Days 5 (d5) or 7 (d7) of the
estrous cycle in Nellore cows.
those treated on Day 5 (45.7% vs. 23.7%, respectively, Table 1).
Increasing PGF2a dose led to greater (P < 0.01) estrus
expression (17.7%, 34.5%, and 51.3%, respectively, Table 1).
Moreover, when the association between the relative drop
in circulating P4 concentration between 0 and 24 hours
after PGF2a and estrous behavior was analyzed, there was
a positive relationship between relative drop in P4 and
estrous behavior, independent of dose of PGF2a (Fig. 2A) or
day of treatment (Fig. 2B). In addition, the relative drop in
concentration to PGF2a doses (12.5, 25, or 50 mg of dinoprost tromethamine)
and Days 5 (d5) or 7 (d7) of the estrous cycle. (A) Probability of estrus for each
PGF2a dose: 12.5 mg of PGF2a (probability of estrus ¼ exp [2.4129 þ 0.0250 
drop P4]/1 þ exp [2.4129 þ 0.0250  drop P4]; P ¼ 0.0003); 25 mg of PGF2a
(probability of estrus ¼ exp [2.2585 þ 0.0307  drop P4]/1 þ exp
[2.2585 þ 0.0307  drop P4]; P  0.0001); and 50 mg of PGF2a (probability of
estrus ¼ exp [1.6592 þ 0.0276  drop P4]/1 þ exp [1.6592 þ 0.0276  drop
P4]; P ¼ 0.0005). (B) Probability of estrus for each day of the estrous cycle: Day
5 (Probability of estrus ¼ exp [2.1521 þ 0.0285  drop P4]/1 þ exp
[2.1521 þ 0.0285  drop P4]; P  0.0001). Day 7 (Probability of estrus ¼ exp
[2.4592 þ 0.0336  drop P4]/1 þ exp [2.4592 þ 0.0336  drop P4];
P < 0.0001). Relative drop in P4 concentration ¼ 100  (P4 concentration at
24 hours after PGF2a  100)/P4 concentration at 0 hours (at PGF2a). a,bEqua-
tions of 12.5 and 50 mg of PGF2a were different (P ¼ 0.0164).
P4 concentration from 0 to 24 hours after PGF2a with
50 mg of PGF2a increased (P ¼ 0.0164) the probability of
estrus behavior than 12.5 mg of PGF2a.
Diameter of the CL at the time of PGF2a was greater
(P < 0.01) on Day 7 than that on Day 5 (15.3  0.20 vs.

Table 1
Main effects of doses of PGF2a (12.5, 25.0, or 50.0 mg dinoprost tromethamine) or days of the estrous cycle (Day 5 or Day 7) of reproductive variables of
Nellore cows.

Variables Day Dose P-value

Day 5 Day 7 12.5 25.0 50.0 Day Dose DD 12.5  25 12.5  50 25  50
n 161 162 108 107 108 d d d d d d
Estrus detection, % 24 46 18 35 51 <0.01 <0.01 0.63 0.04 0.05 0.05
CLb, mm 13 15 14 14 14 <0.01 0.29 0.50 d d d
CLa, mm 12 9 12 11 10 <0.01 <0.01 0.80 0.03 <0.01 0.24
L1.0, % 30 77 39 52 70 <0.01 <0.01 0.65 0.03 <0.01 0.01
L0.5, % 17 48 23 34 40 <0.01 0.02 0.40 0.15 <0.01 0.21
Partial luteolysis, % 57 19 49 37 28 <0.01 <0.01 0.26 0.03 <0.01 0.23
No luteolysis, % 12 4 12 10 2 0.96 0.96 0.95 d d d

Partial luteolysis, circulating P4 concentration was less than 1.0 ng/mL 24 h after PGF2a treatment, but it was greater than 1.0 ng/mL 48 h after treatment.
Abbreviations: CLa, CL diameter at 96 h after treatment with PGF2a; CLb, CL diameter before PGF2a treatment (Day 5 or Day 7); Day 5, fifth day of the estrous
cycle; Day 7, seventh day of the estrous cycle; D  D, interaction between days and doses; L0.5, luteolysis 0.5 (cows with P4 < 0.5 ng/mL at 72 h after PGF2a);
L1.0, luteolysis 1 (cows with P4 < 1.0 ng/mL at 72 h after PGF2a); n, number of cows in each treatment.
 M.V.C. Ferraz Junior et al. / Theriogenology 86 (2016) 1268–1274
12.5  0.18 mm; respectively, Table 1), but did not vary
between cows treated with different PGF2a doses on the
same day of the estrous cycle. Diameter of the CL at
96 hours after PGF2a (CLa) was greater (P < 0.01) on Day 5
than Day 7 and cows that received 12.5 mg of PGF2a
(11.6  0.36 mm) had greater (P < 0.01) CLa diameter than
cows that received 25.0 or 50.0 mg of PGF2a (10.7  0.37
and 10.1  0.37 mm, respectively, Table 1).
The incidence of luteolysis was greater (P < 0.01) on Day
7 than that on Day 5 (L1.0–77.2% vs. 30.4%; L0.5–47.6% vs.
17.3%, respectively, Table 1) and increasing the dose of
PGF2a increased (P < 0.01) L1.0. For L0.5, incidence of
luteolysis was greater (P < 0.01) with 50.0 mg of PGF2a than
that with 12.5 mg of PGF2a (39.8% vs. 23.0%, respectively,
Table 1). The incidence of partial luteolysis was greater
(P < 0.01) on Day 5 (57.1%) than that on Day 7 (19.1%) of the
estrous cycle. A greater percentage of cows treated with
12.5 mg of PGF2a had partial luteolysis (Table 1).
Circulating P4 concentration decreased 24 hours after
PGF2a treatment, on Day 5 and Day 7 of the estrous cycle,
and the magnitude of decrease was also influenced by the
dose of PGF2a on both days. At 48 and 72 hours after PGF2a
treatment, there was a rebound on circulating P4 concen-
trations in cows treated on Day 5 of the estrous cycle with
any dose of PGF2a and in cows treated on Day 7 of the
estrous cycle with 12.5 mg of PGF2a (Fig. 3A, B).
Either at Day 5 or Day 7 of the estrous cycle, there was
no difference between PGF2a treatments on circulating P4
concentration if only cows with partial luteolysis were
considered in the analyses (Fig. 3C, D). In addition, there
was no difference in P4 concentration in cows with com-
plete luteolysis that had been treated either on Day 5 or
Day 7 of the estrous cycle (Fig. 3E, F).
Considering cows that showed estrus, six of them were
classified as having partial luteolysis. All of these cows were
from the Day 5 group: one was from Day 5/12.5PGF2a, one
from Day 5/25.0PGF2a, and four from Day 5/50.0PGF2a
treatment. This group of cows had circulating P4 concen-
tration above 1 ng/mL at 0 hours. Progesterone concentra-
tion dropped below 1 ng/mL at 24 and 48 hours after PGF2a
administration, however, P4 rebounded above 1 ng/mL at
72 hours after PGF2a.
4. Discussion
In the present study, the efficacy to induce luteolysis
of varying doses of PGF2a that are used in standard estrus
synchronization protocols was evaluated at relatively
early stages (Day 5 and Day 7) of the estrous cycle. Pri-
mary findings from this experiment were that on Day 5,
even the greatest dose of PGF2a was only effective in
approximately 50% of cows. Furthermore, on Day 7, it
appears that the 12.5-mg PGF2a dose was insufficient to
ensure complete luteolysis in most cows. Each of these
findings has important implications for synchronization
of estrus in Nellore females, and the most appropriate
approaches to take as new programs are investigated and
developed.
Multiple responses support that there exist significant
variation on Day 5 versus Day 7 and between doses on
both Day 5 and Day 7. More cows were detected in estrus
1271
when treated on Day 7 versus Day 5, and incidence of
estrus increased with dose of PGF2a. Variation in incidence
of estrus was strikingly similar to the variation between
day of the cycle and dose in the incidence of partial or
failed luteolysis. Similar relationships have been previ-
ously reported in Nellore [14]. The occurrence of estrus
in cows treated on Day 7 was approximately 10 hours
earlier than that in cows treated on Day 5. This was likely
the result of more fully developed dominant follicles in
the ovary when PGF2a was given on Day 7 versus Day 5
[15–17].
As expected, diameter of the CL present at the time of
PGF2a was less on Day 5 than that on Day 7, as it has been
demonstrated that in Bos indicus cattle, the CL continues to
increase in diameter up to Day 9 of the estrous cycle [18].
The inverse relationships between diameter of CL 96 hours
after PGF2a with day of the estrous cycle and dose of PGF2a
lend further evidence to those derived from using P4 con-
centrations. According to Cuervo-Arango et al. [7], when
luteolysis is complete, the CL regresses and become almost
imperceptible within 4 days. Moreover, when luteolysis is
incomplete or partial, diameter of the CL declines in most
animals during the first 12 hours after PGF2a treatment
[4,7]. In Gir cattle, structural regression of the CL occurred
at a rate of 0.36  0.04 cm2 per day [18] between Days 16
and 21 of the estrous cycle.
Data of our work also showed that higher doses
(50.0 mg) of PGF2a increased luteolysis rate in approxi-
mately 15% and 10% points on Day 5 and Day 7 of the
estrous cycle, respectively, when compared with the rec-
ommended dose (25.0 mg). These results corroborate data
from others [8], who described an increase in 20% points of
luteolysis when double dose of d-cloprostenol was
administrated 130 hours after ovulation. The greater
luteolytic effect of doubling the dose of PGF2a in FTAI pro-
tocols was also shown by Brusveen et al. [19] in dairy cows
submitted to a standard 7-day Ovsynch protocol. Complete
luteolysis was observed in 95.6% of cows treated with twice
the recommended dose, whereas the single dose resulted
in 84.6% of cows presenting complete luteolysis. The
authors also have shown that doubling the dose of PGF2a,
increased conception rates.
Our data corroborate the idea that the transition of the
CL, from being refractory to being responsive to PGF2a, oc-
curs around the fifth to sixth day on the estrous cycle [4], as
luteolysis was more evident at Day 7 compared with Day 5.
However, treatment with PGF2a was able to induce w30% of
luteolysis on Day 5. These results contrast with data pre-
sented by Nascimento et al. [5], who did not detect complete
luteolysis in any of the cows treated with one or two doses of
PGF2a 5 days after GnRH administration in nonlactating
Holstein cows. However, other authors showed that in cows
treated with PGF2a 120 hours after ovulation, it was possible
to induce luteolysis in w50% of the cows [4,7]. The lack of CL
regression after PGF2a challenge at the beginning of diestrus
has been extensively studied in ruminants [1,3,6,8], but
reasons are not completely understood. It has been
demonstrated that PGF2a exerts actions on early and mature
CL [9]. The lack of PGF2a-induced luteolysis in the early CL
may be due to specific changes in gene expression that may
prevent intraluteal PGF2a production [9]. Other possible
1272 M.V.C. Ferraz Junior et al. / Theriogenology 86 (2016) 1268–1274

Fig. 3. Progesterone (P4) concentration at 0, 24, 48, and 72 hours after treatment with PGF2a (12.5, 25, or 50 mg dinoprost tromethamine) on Days 5 or 7 of the
estrous cycle. (A) and (B) represent all cows in their respective groups (treated with PGF2a on Day 5 or 7 of the estrous cycle). (C) and (D) represent only cows without
complete luteolysis on Days 5 and 7 of the estrous cycle. (E) and (F) represent only cows with complete luteolysis on Days 5 and 7 of the estrous cycle. P valuedD,
dose of PGF2a; H, hours after PGF2a treatment; D  H ¼ interaction between dose and hour. Cow numbersd(A) 12.5-mg PGF2a: n ¼ 54; 25.0-mg PGF2a: n ¼ 54; and
50.0-mg PGF2a: n ¼ 53; (B) 12.5-mg PGF2a:n ¼ 54; 25.0-mg PGF2a: n ¼ 53; and 50.0-mg PGF2a: n ¼ 55; (C) 12.5-mg PGF2a: n ¼ 45; 25.0-mg PGF2a: n ¼ 41; and
50.0-mg PGF2a: n ¼ 26; (D) 12.5-mg PGF2a: n ¼ 21; 25.0-mg PGF2a: n ¼ 10; and 50.0-mg PGF2a: n ¼ 6; (E) 12.5-mg PGF2a: n ¼ 9; 25.0-mg PGF2a: n ¼ 13; and 50.0-mg
PGF2a: n ¼ 27; (F) 12.5-mg PGF2a: n ¼ 33; 25.0-mg PGF2a: n ¼ 43; and 50.0-mg PGF2a: n ¼ 49. **The behavior of responses to treatments with 25.0 and 50.0 mg of
PGF2a is similar, resulting in one line overlaying another.
 M.V.C. Ferraz Junior et al. / Theriogenology 86 (2016) 1268–1274
explanation to this resistance is the unavailability of
endothelin-1 [20] and the increase of prostaglandin dehy-
drogenase [21] in immature CL, compared with mature CL.
Moreover, it is possible that other factors such as experi-
mental design, breed, age, and feeding influence the luteo-
lytic response to PGF2a.
Several studies have shown that if PGF2a is adminis-
trated early in the cycle, it does not induce complete
luteolysis, it induces partial luteolysis [4–8]. The partial
luteolysis is characterized by a relatively rapid decrease in
circulating P4 concentration, followed by a rebound 48 to
72 hours after PGF2a treatment [4–8]. In our study, most
cows that did not have complete luteolysis, had partial
luteolysis, especially when treated on Day 5. Despite that,
partial luteolysis also occurred in cows treated on Day 7.
Increasing the dose of PGF2a either on Day 5 or Day 7 did
not affect circulating P4 concentration at 24, 48, and
72 hours after PGF2a administration in cows with partial
luteolysis. Data from other studies may help to explain this
observation. It is possible that initially StAR and NR5A1, that
are responsible for cholesterol transport, have their func-
tion diminished due to the action of PGF2a, but some
essential steroidogenic genes as LHCGR, CYP11A1, and
P450scc can hold their expression capacity [22]. Finally,
StAR can restore its function, and thus P4 production is
retrieved. It is also likely that with partial luteolysis, the
luteolytic cascade is interrupted before the production of
intraluteal PGF2a that is important to achieve complete
luteolysis [23]. Our study also showed that a small group of
cows had no change in circulating P4 24 hours after PGF2a
administration.
Achieving timely and complete luteolysis is crucial, as it
has been reported that cows exhibiting P4 concentration
greater than 0.1 [13] or greater than 0.5 ng/mL [11,24] at the
time of AI may have fertility impairment. When we were
more stringent and analyzed the L0.5, it was observed that
there was luteolysis in only 50% of cows treated with the
double dose of PGF2a on Day 7, which was the treatment
with the greatest response.
The six cows that showed estrus but were classified
with partial luteolysis had P4 concentration around
1.5 ng/mL on Day 5. After that, these cows remained with P4
concentration around 0.5 ng/mL for 2 days. We speculate
that these windows of low P4 concentration were enough
to allow the development of dominant follicle, and there-
fore, an estradiol peak. On the other hand, it is also possible
that we may have had false positive results at estrus
observation. According to Garcia [25], 5.8% of crossbred
Nellore cows showed signs of estrus while having elevated
circulating P4 concentration.
4.1. Conclusions
The use of PGF2a on Day 7 of the estrous cycle was more
efficient to induce luteolysis than that if given on Day 5 of
the estrous cycle. Furthermore, increasing the PGF2a dose
from 12.5 mg to 50.0 mg increased the percentage of cows
that experienced complete luteolysis. Consideration of
these data is crucial to interpret responses and designing
new estrus synchronization protocols. It is obvious that
failure of complete luteolysis is a potential contributor to
1273
variation in pregnancy rates in estrous synchronization
protocols and especially in FTAI programs.
Acknowledgments
This work was financially supported by the Fundação de
Amparo à Pesquisa do Estado de São Paulo (FAPESP 2012/
01345-9). The authors also thank Experimental Station
Hildegard Georgina Von Pritzelwitz (Fundação de Estudos
Agrários Luiz de Queiroz) for allowing the use of their cattle
and facilities during this study.
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