Theriogenology 79 (2013) 882889

Contents lists available at SciVerse ScienceDirect

Theriogenology
journal homepage: www.theriojournal.com

Evaluation of three synchrony programs for pasture-based dairy heifers

S. McDougall a, *, F.M. Rhodes b, C.W.R. Compton a
a
b

Cognosco, Anexa Animal Health, Morrinsville, New Zealand

Hamilton, New Zealand

a r t i c l e i n f o

a b s t r a c t

Article history:
Received 3 June 2012
Received in revised form 20 December 2012
Accepted 23 December 2012

The objective was to evaluate the efcacy and economic benets of three synchrony
programs in 1137 heifers from 10 pasture-based dairy herds. Heifers were randomly
assigned to one of three treatments within each herd on Day
13 (Day 0 start of the
breeding program). They were treated with: (1) PGF2a on Days
13 and
2, with AI after
detection of estrus between Days 0 and 3 (Double PG); (2) GnRH, PGF2a, and GnRH on
Days
9,
2, and 0, respectively, with placement of an intravaginal progesterone
(P4)-releasing device between Days
9 and
2, and set time AI on Day 1 (GPG P4); or
(3) same as the GPG P4 group but with the set time AI on Day 0 (Cosynch P4). Plasma
P4 concentrations were determined on Days
20 and
13 to determine pubertal status.
The Cosynch P4 treatment had a higher (P < 0.05) conception rate to AI (57% vs. 47%
vs. 48% for Cosynch P4, GPG P4, and Double PG, respectively), 21-day in-calf rate (76%
vs. 72% vs. 63% for Cosynch P4, GPG P4, and Double PG), and a shorter median
interval from the start of the breeding program to conception (0, 14, and 19 days for
Cosynch P4, GPG P4, and Double PG). Heifers that had reached puberty before
breeding, compared with those that had not, had higher (P < 0.05) in-calf rates to AI (53%
vs. 47%) at 21 days (74% vs. 64%) and at 42 days (91% vs. 84%). Pubertal status was
associated with herd, breed, age, and body condition score at the start of mating (P <
0.05). A partial budget model demonstrated that, compared with the Double PG program,
there was an economic benet from the Cosynch P4 (mean, NZ$25.73; 95% condence
interval, 2.9950.69), but not the GPG P4 program (mean, NZ$
0.65; 95% condence
interval,
21.87 to 21.58). We concluded that the Cosynch P4 program resulted in the
highest fertility and economic benet of the three programs evaluated, and that reproductive response was affected by pubertal status.
2013 Elsevier Inc. All rights reserved.

Keywords:
Heifer
Synchrony
Ovsynch
Cosynch
Progesterone
Prostaglandin

1. Introduction
Synchronization of estrus and ovulation of dairy heifers,
in combination with AI, facilitates improving the rate of
genetic gain by shortening the generation interval and
obtaining replacements from dams with the highest
genetic merit in a herd [1]. However, the use of AI in dairy
heifers has historically been limited in New Zealand. For
example, in 2006 and 2007 only 118,226 yearlings were
mated using AI compared with 2,904,667 cows, from a total
* Corresponding author. Tel.: 64 7 889 5159; fax: 64 7 889 3681.
E-mail address: smcdougall@anexa.co.nz (S. McDougall).
0093-691X/$ see front matter 2013 Elsevier Inc. All rights reserved.
http://dx.doi.org/10.1016/j.theriogenology.2012.12.013

population of 5.3 million dairy cattle [2]. The practicalities

of inseminating heifers after either daily estrous detection
or some form of synchrony might be barriers to uptake of
the technology.
Various synchrony programs for dairy heifers have
previously been evaluated in pasture-based systems [36].
However, these programs generally included the use of
estradiol, which can no longer be used in food-producing
animals because of the European ban, or involved detection of estrus. Therefore evaluation of new programs was
required, including those that did not require estrous
detection and were practical under the extensive pasturebased management systems of New Zealand.

S. McDougall et al. / Theriogenology 79 (2013) 882889

Use of xed-time insemination (FTAI) removes the

requirement for estrous detection, but conception rates
have been reported to be signicantly lower using FTAI
after two injections of PGF2a 10 to 14 days apart (Double
PG) than for dairy heifers inseminated on detection of
estrus [7]. These low conception rates were attributed to
variations in time to estrus and ovulation depending on the
stage of the estrus cycle at PGF2a administration, and the
lack of efcacy of PGF2a in prepubertal animals [8,9].
Optimal synchrony of estrus and ovulation requires
control of follicle development and luteal function [10].
This can be achieved in dairy cows using programs
involving GnRH injections 9 days apart with PGF2a 2 days
before the nal GnRH treatment, with FTAI 16 hours after
the nal GnRH treatment (GPG or Ovsynch), but such
programs used in heifers do not achieve conception rates
comparable with AI after detection of estrus, because of
a lack of synchrony of estrus [11,12]. Insemination might
also occur coincident with the nal GnRH treatment
(Cosynch). The Cosynch program reduces the number of
handlings of cattle required, but has been reported to result
in lower conception rates compared with Ovsynch [13].
However, in beef heifers, a Cosynch program resulted in
pregnancy rates superior to a Double PG program [14].
Addition of progesterone (P4) to GPG programs also
resulted in signicantly increased conception rates after
FTAI in beef heifers [15], and improved synchrony of estrus
in dairy heifers [16].
Heifers that are prepubertal at the commencement of
synchrony programs are less likely to exhibit estrus [17], and
have lower conception and pregnancy rates than postpubertal heifers [18]. In prepubertal beef heifers, inclusion of
P4 in the synchrony program signicantly increased submission and pregnancy rates [17], and conception rates [19].
The aim of the current study was to evaluate the efcacy
and economic benets of three synchrony programs in
heifers from pasture-based dairy herds. The programs were
GPG or Cosynch, both with the addition () of P4, and
Double PG. It was hypothesized that more heifers would
conceive to AI and be pregnant by Day 21 of the breeding
program after a GPG P4 program than after Double PG,
and that conception rate to AI would be noninferior after
Cosynch P4 compared with GPG P4.
2. Materials and methods
2.1. Heifers and treatments
The study was conducted after approval from the Animal Ethics Committee of AgResearch Ruakura, Hamilton,
New Zealand.
Dairy heifers (N 1137) from 10 herds (mean  SD,
114  26 per herd) were enrolled on one calendar day for
each herd. All herds were spring-calving and located
within the Waikato region of New Zealand. Blood samples
(10 mL) were drawn from the tail vein into an evacuated
glass tube containing lithium heparin as an anticoagulant
(Vacutainer; Becton Dickson, Franklin Lakes, NJ, USA) on
Days
20 and
13 (where Day 0 start of the breeding
program) for evaluation of P4 concentration in plasma by
RIA (Coat-a-Count; DPC Corporation, Los Angeles, CA,

883

USA). Heifers were dened as prepubertal if the P4

concentration in both samples was <1.0 ng/mL, and
postpubertal if P4 concentration in one or both samples
was 1.0 ng/mL.
On Day
13, the body condition score (BCS) of each
heifer was assessed on a one to 10 scale [20], and tail paint
applied to aid estrus detection. Heifers were randomly
assigned, within sequentially presented blocks of three
heifers, to one of three treatment groups. Different color
tail paints were used for identication of each treatment
group, but all heifers within a herd were managed as
a single management group.
The three treatments were:
 Double PG: Heifers were treated with cloprostenol
(Ovuprost 500 mg im; Bomac Laboratories Ltd., Manukau
City, New Zealand) on Days
13 and
2, with AI upon
estrus detection between Days 0 and 3 (N 380; 33%);
 GPG P4: Heifers were treated with an intravaginal
P4-releasing device (Cue Mate; Bomac Laboratories Ltd.)
from Days
9 to
2, and gonadorelin (Ovurelin 100 mg
im; Bomac Laboratories Ltd.) on Day
9, 500 mg cloprostenol im on Day
2, and 100 mg gonadorelin on Day 0,
with FTAI on Day 1 (N 383; 34%); and
 Cosynch P4: Heifers were treated in the same manner
as for GPG P4 but with FTAI coincident with the nal
gonadorelin treatment (N 374; 33%).
The timing of the treatments was such that the second
cloprostenol treatment for the Double PG group coincided
with the cloprostenol treatment of the other two groups.
Treatments were given between 9:00 AM and 1:00 PM.
All heifers were assessed once daily from Days 0 to 3 for
loss of tail paint while yarded for drafting for AI. Herd
owners decided which of the heifers in the Double PG
group were in estrus and were to be inseminated each day
based on removal of tail paint or their own observations on
pasture. When a heifer had been inseminated, no more
estrous observations were conducted.
Experienced AI technicians attended each herd at
approximately midday on Days 0 to 3. In seven herds, one
technician undertook all inseminations, whereas in the
remaining three herds, two technicians undertook inseminations. Where two technicians were used, they undertook inseminations on each day of the program. Heifers in
the Double PG group detected in estrus between Days 0 and
3 were inseminated on the day of detection. Heifers in the
GPG P4 group in estrus at the time of the second GnRH
injection (Day 0) were recorded as in estrus, but not
inseminated until Day 1.
Frozen semen from 40 sires was used for AI in the study
(one to four sires were used in each treatment group). After
the last AI on Day 3, bulls were placed with the heifers for
a mean (SD) total duration of 80  10 days. The number of
bulls required for each management group was calculated
assuming that 50% of the heifers would conceive to rst
service and that those not conceiving would return to
estrus over a 5-day period on average 21 days later. Sufcient bulls were introduced such that no more than three
services per bull per day would occur on average.

884

S. McDougall et al. / Theriogenology 79 (2013) 882889

Pregnancy diagnosis was conducted in all herds

(mean  SD) 69  2 days and 109  5 days after the start of
the breeding program. In seven herds, additional pregnancy diagnosis took place 139  6 days after the start of
the breeding program. The stage of gestation, in days, was
estimated at each examination if a heifer was diagnosed
pregnant.
The date of birth and breed of individual heifers was
retrieved from an electronic database (LIC, Hamilton, New
Zealand). These data were used to calculate the age at the
start of the breeding program (mean  SD 14.3  0.4
months), and to classify heifers as Friesian (N 575; 51%),
Jersey (N 159; 14%), or other (N 403; 35%).
2.2. Denitions and statistical methods
There were six primary outcome variables in this study.
The in-calf to AI rate was dened as the proportion of
enrolled heifers that conceived to AI. The conception rate to
AI was dened as the proportion of heifers conceiving to AI
and was calculated only for heifers that were submitted for
AI between Days 0 and 3. It was possible that some heifers
in the Double PG group might have conceived to the rst
natural mating after the introduction of bulls, but natural
matings were not recorded, and so these heifers were
excluded from this analysis. The 21-day and 42-day in-calf
rates were dened as the proportion of enrolled heifers
pregnant at 21 or 42 days from the start of the breeding
program, respectively. The nal in-calf rate was dened as
the proportion of heifers present at the nal pregnancy test
that were pregnant at the nal pregnancy test. Finally, the
median interval in days from the start of the breeding
program to conception was calculated by survival analysis
methods. For heifers in the GPG P4 group, the reproductive performance measures were calculated using the
rst day of AI as the start point (Day 1 in the study), so as
not to bias the results for this group compared with those
in the other two groups that started 1 day earlier (Day 0 in
the study).
Potential predictor variables evaluated were treatment
group, herd of origin, age in months at the start of breeding
(categorized into quartiles), pubertal status at commencement of the study, breed category, observation of estrus,
and BCS (categorized into quartiles).
Balance of treatment groups for age, breed, pubertal
status, and BCS of heifers was compared using chi-square
tests. Differences between treatment groups in the categorical outcome variables were compared using two-sided
Pearson chi-square test with Holm adjustment for multiple
testing [21] to control the overall type I error rate. Multivariable logistic regression using a forward stepwise addition method was used to model the effect of treatment
group, and other variables associated by univariate analysis
(P < 0.2), on the outcomes. Treatment group and herd of
origin were tted as xed effects and forced into each
model. Other variables were tested for signicance and as
potential confounders and included if their Wald test
probability value was P < 0.05, or if their inclusion changed
the coefcients for the treatment effects by 15% or more.
Interactions between treatment groups and other signicant variables were also tested. Preliminary nal models

were checked by plotting the deviance residuals against the

tted values and the Cook statistics against the standardized leverages, and if no values were of concern, were
declared nal. The mean effects of the other nontreatment
variables in the nal models were also estimated to
describe the change in outcome (on a percentage scale)
associated with that variable. The noninferiority hypothesis
was tested using differences between the means and the
condence intervals of the differences estimated from the
nal model, with the other included variables set to their
median value. The effect on the time to conception of
treatment group and potential confounding variables were
tested by KaplanMeier survival analysis using the log-rank
test, and survival curves plotted.
A variable of secondary interest was pubertal status as
dened by plasma P4 concentrations. Associations were
evaluated between pubertal status and herd of origin,
breed, and age and BCS at Day
13 using chi-square analysis with adjustment for multiple testing, as previously
described. The pubertal status (post vs. prepubertal) of
heifers was modeled by multiple logistic regression models
and effects of signicant variables estimated as previously
described.
2.3. Power statistics
The rst hypothesis was that more heifers would
conceive to AI and be pregnant by Day 21 of the breeding
program after a GPG P4 program than after Double PG. It
was assumed that submission rates would be 70% and
100%, and conception to AI rates would be 60% and 55%, for
the Double PG and GPG P4 groups, respectively. Thus it
was expected that approximately 42% and 55% of heifers
would be pregnant to AI in the Double PG and GPG P4
groups, respectively. To detect this 13% difference,
approximately 250 heifers per treatment group were
required (a 0.05; a 0.2). To allow for loss to follow-up,
approximately 300 heifers per treatment group were
enrolled.
The second hypothesis was that pregnancy rates by Day
21 would be noninferior for the Cosynch P4 compared
with the GPG P4 group. It was assumed that the
conception rate of the two groups would be the same (55%)
and an acceptable difference between the two groups
would be 10% (a 0.05; b 0.2). This required 420 heifers
per treatment group (Pass 2008; www.ncss.com).
2.4. Partial budget
A partial budget model was prepared to estimate the
economic effect of synchronization of estrus of heifers using
the GPG P4 or Cosynch P4 programs compared with the
Double PG program. The model was built using input data
and distributions from the current study, other published
data, or the opinions of the authors (Table 1), and the
method of stochastic Monte Carlo simulation in the Microsoft Excel add-in @Risk (Version 5; Palisade Corporation,
Ithaca, NY, USA). This simulation modeling takes account of
the variability of model inputs by assigning each a specic
distribution, and generates a distribution for the outcomes
from which mean economic effects are estimated.

S. McDougall et al. / Theriogenology 79 (2013) 882889

885

Table 1
Assumptions used to develop a stochastic partial budget model (in New Zealand $) in the Microsoft Excel add-in @Risk to compare the economic benet of
estrus synchrony with articial breeding of pasture-grazed dairy heifers using either GPG P4a or Cosynch P4b with a Double PGc treatment program.
Income variables

Mean

Distribution

Extra days in milk GPG P4d

Extra days in milk Cosynch P4d
Mean milk production per day (kg milk solidse)
Milk payment next season ($ per kg milk solidse)
Percentage in calf to AI, Double PG
Percentage in calf to AI, GPG P4
Percentage in calf to AI, Cosynch P4
Percentage calves female
Extra AI heifer calves, GPG P4d
Extra AI heifer calves, Cosynch P4d
Male calf value
AI calf value (4 days of age)

3.9
6.4
1.2
5.50
31%
47%
57%
50%
8%
13%
$20.00
$200.00

RiskNormal(3.9, 1.5)
RiskNormal(6.4, 1.5)
RiskNormal(1.2, 0.12)
RiskUniform(5.65, 5.75)
RiskLogistic(0.31, 0.0136)
RiskLogistic(0.47, 0.0136)
RiskLogistic(0.57, 0.0135)
RiskBinomial(100, 0.5)

Cost variables

RiskUniform(15, 25)
RiskUniform(150, 250)

Mean

Drug and treatment costs per head

Drug costs, Double PG
Drug costs, GPG + P4
Drug costs, Cosynch + P4
Veterinary fees, Double PG
Veterinary fees, GPG + P4
Veterinary fees, Cosynch + P4

$7.36
$40.00
$40.00
$6.00
$11.00
$11.00

Overhead costs per head

Mean

Average number of heifers per group

Number of heifer yardings, Double PG
Number of heifer yardings, GPG P4
Number of heifer yardings, Cosynch P4
Grazier yarding fees
Percentage of graziers charging yarding fees
Visit and yarding fees plus travel, Double PG
Visit and yarding fees plus travel, GPG P4
Visit and yarding fees plus travel, Cosynch P4
Difference in overhead costs per head, GPG P4d
Difference in overhead costs per head Cosynch P4d

50
5
4
3
$50.00
25%
$222.50
$290.00
$277.50
$1.35
$1.10

Distribution

RiskUniform(25, 75)
RiskTriang(0.2, 0.25, 0.3)

Distribution is the @Risk formula used to model variability; otherwise the mean value was used. Income variables are shown.
a
PGF2 on Days
13 and
2 with AI on detection of estrus from Day 0 to 3 of the breeding program.
b
GnRH, PGF2 , and GnRH on Days
9,
2, and 0 of the breeding program, respectively, with insertion of an intravaginal progesterone-releasing insert for
7 days commencing on Day
9, and xed time AI on Day 1.
c
GnRH, PGF2 , and GnRH on Days
9,
2, and 0 of the breeding program, respectively, with insertion of an intravaginal progesterone-releasing insert
for 7 days commencing on Day
9, and xed time AI on Day 0.
d
Compared with Double PG.
e
Milk solids kg of protein plus kg of fat.

In seasonal calving systems with a xed start and end of

lactation, the maximum number of days in milk occurs
when an animal calves at the start of the calving program
(having conceived at the start of the breeding program) and
each day of delay to conception results in one day less in
milk. Hence, the additional number of days in milk in the
lactation after synchrony for the GPG P4 or Cosynch P4
heifers, relative to the Double PG heifers, was modeled by
subtracting the interval from the start of breeding to the
date of conception from the duration of the breeding
program for each heifer, and including the mean and SEM
of the differences between treatment groups in the model.
The economic benets were assessed in terms of extra
milk production, calculated from the estimated value of
milk solids (kg of milk protein plus kg of milk fat; the basis
of payment in the New Zealand dairy industry), mean daily
production of milk solids, and extra days in milk, plus the
value of additional AI calves, calculated from the estimated
percentage of extra female AI calves born and the value
of these calves at 4 days of age. Additional costs were

calculated from estimated treatment costs based on drug

costs and vet fees, plus management costs because of
yarding of heifers.
The means and 95% condence intervals of the distributions of the economic benet of each comparison were
reported and the treatment comparison declared signicantly different if the 95% condence interval of the
difference did not include zero. Spearman rank correlation
coefcients were examined to determine the variables with
greatest inuence on the net economic benet.
Data were managed in a custom-built Microsoft Access
database (Microsoft, Redmond, WA, USA), and analysis
conducted using R (Version 2.8.1; R Development Core
Team, 2008). Statistical signicance was declared for test
results with P < 0.05.
3. Results
Nine heifers were removed from the analysis because
they were infertile (freemartins, N 6), pregnant and

886

S. McDougall et al. / Theriogenology 79 (2013) 882889

aborted within 1 day after treatment with PGF2a (N 1), or

were missing at the nal treatment with PGF2a (N 2),
leaving data for analysis from 1137 heifers. Six heifers were
not present for the nal pregnancy diagnosis and were not
included in results for the nal in-calf rate.
Mean BCS of heifers at enrolment was 4.6, with 90% in
the range of 4.0 to 5.5 (inclusive). Paired plasma samples
were available from 1123 heifers; 671 heifers (60%) had
plasma P4 concentrations 1.0 ng/mL in one or both
samples.
There were no differences between treatment groups in
age category (P 0.71), breed category (P 0.81), pubertal
status (P 0.87), or BCS category (P 0.92).
3.1. Detection in estrus
A total of 248 of 380 heifers (65%) in the Double PG
group were detected in estrus by farmers and submitted for
AI over Days 0 to 3. The percentage of heifers detected in
estrus in this group was greater for those that were postcompared with prepubertal (87.3% vs. 34.2%; P < 0.01).
On Day 0, 107 of 380 (28%), 183 of 383 (48%), and 178 of
374 (48%) of heifers were observed at the time of yarding to
have their tail paint disturbed for the Double PG, GPG P4
and Cosynch P4 groups, respectively (P < 0.001).
3.2. Conception to AI
Results for the in-calf to AI rate and conception rate to AI
for the three treatment groups are summarized (Table 2).
For both outcomes, heifers in the Cosynch P4 group
achieved higher values than those in the GPG P4 group
(P < 0.05). In-calf to AI rate was also higher (P < 0.01) and
conception to AI tended to be higher (P 0.069) for heifers
in the Cosynch P4 group than for heifers in the Double PG
group.
The Cosynch P4 group was declared noninferior to the
GPG P4 group because the lower 95% condence interval
of the difference in conception rate to AI was greater than

0.10, the a priori dened acceptable difference (difference
(Coynsch P4)
(GPG P4) 0.11; 95% condence interval,
0.020.19). At the univariate level, the in-calf rate to AI

varied among herds and was higher in pubertal compared

with prepubertal heifers (53% vs. 47%; P 0.05), but only
treatment group and herd were signicant in the nal
model. Conception rate to AI also varied among herds
(range, 37%63%) and was higher in heifers from which tail
paint was removed 1 to 48 hours before AI compared with
0 to 48 hours after AI (266/497 [53.5%] vs. 73/186 [39.2%];
P < 0.001).
3.3. Pregnancy rates
Results for the 21-day, 42-day, and nal in-calf rates for
the three treatment groups are summarized (Table 2).
Heifers in the Cosynch P4 group had higher 21- and
42-day in-calf rates than heifers in the Double PG group
(P < 0.05), but did not differ from those in the GPG P4
group. The 21-day in-calf rate varied among herds (range,
53%80%). In the nal models, heifers that had reached
puberty before breeding, compared with those that had
not, had higher in-calf rates at 21 days (74% vs. 64%; P <
0.001) and 42 days (91% vs. 84%; P 0.03).
The nal in-calf rates were not different between
treatment groups, but varied among herds. For all nal
models, there were no signicant herd by treatment
interactions or pubertal status by treatment interactions.
Survival curves for the interval from the start of the
breeding program to conception differed between the
treatment groups (P < 0.001; Fig. 1). The median interval
from the start of breeding to conception was shorter for the
heifers in the Cosynch P4 than the GPG P4 and Double
PG groups (P < 0.05), and tended to be shorter for heifers in
the Cosynch P4 group than the GPG P4 group (P 0.06;
Table 2).
3.4. Pubertal status
Overall, 671 of 1123 heifers (60%) were categorized as
postpubertal, but there was considerable variation among
herd of origin (range, 9%93%). Factors associated with
pubertal status were herd of origin (P < 0.001), breed
category (P < 0.001), age (P 0.002), and BCS at the start of
the breeding program (P < 0.001; Table 3).

Table 2
Number (%) of dairy heifers conceiving to AI or pregnant after three treatment protocols for synchronization of estrous and ovulation.
Treatment
group

In-calf to AIa

Conception
rate to AIb

21-Day
in-calf ratec

42-Day
in-calf rated

Final
in-calf ratee

Interval to
conception (days)f

Double PGg
GPG P4j
Cosynch P4k

119/380h (31.3)
180/383i (47.0)
213/374l (57.0)

119/248h,i (48.0)
180/383h (47.0)
213/374i (57.0)

238/380h (62.6)
274/383i (71.5)
284/374i (75.9)

313/380h (82.4)
335/383h,i (87.5)
333/374i (89.0)

347/376 (92.3)
352/381 (92.4)
358/374 (95.7)

19h
14h
0i

Proportion of all heifers conceiving to AI.

Proportion of heifers articially bred conceiving to AI.
c
Proportion of all heifers pregnant after 21 days of the breeding program.
d
Proportion of all heifers pregnant after 42 days of the breeding program.
e
Proportion of heifers examined that were pregnant at the end of the breeding program.
f
Median interval from start of breeding to conception.
g
PGF2 on Days
13 and
2 with AI on detection of estrus from Day 0 to 3 of the breeding program.
h,i, and l
Within a column, means without a common superscript differed (P < 0.05).
j
GnRH, PGF2 , and GnRH on Days
9,
2, and 0 of the breeding program, respectively, with insertion of an intravaginal progesterone-releasing insert
for 7 days commencing on Day
9, and xed time AI on Day 1.
k
GnRH, PGF2 , and GnRH on Days
9,
2, and 0 of the breeding program, respectively, with insertion of an intravaginal progesterone-releasing insert for
7 days commencing on Day
9, and xed-time AI on Day 0.
b

S. McDougall et al. / Theriogenology 79 (2013) 882889

887

4. Discussion

Fig. 1. KaplanMeier survival curves of cumulative time to conception from

the start of the breeding program (Day 0) for dairy heifers treated with three
synchronization protocols: (1) PGF2a on Days
13 and
2, with AI after
detection of estrus between Days 0 and 3 (Double PG); (2) GnRH, PGF2a, and
GnRH on Days
9,
2, and 0, respectively, with placement of an intravaginal
progesterone (P4)-releasing device between Days
9 and
2, and set time
AI on Day 1 (GPG P4); or (3) same as the GPG P4 group but with the set
time AI on Day 0 (Cosynch P4). There were differences among treatment
groups (P < 0.001).

3.5. Partial budget

The Cosynch P4 program had an economic benet
compared with the Double PG program, but there was no
benet from the GPG P4 program (Table 4). Additional
income from greater milk yield because of earlier calving
and more AI calves in the Cosynch P4 compared with the
Double PG program accounted for the difference between
the two groups. Costs for the two P4-based treatments were
very similar. The variables most highly correlated with the
net benet of the Cosynch P4 program were the additional days in milk in the subsequent lactation (rs 0.83)
through earlier conception pattern, daily milk production
(rs 0.35), and value of an articially-sired calf (rs 0.3).

Table 3
Percentage of heifers (N 1123) that were postpubertal, based on
progesterone concentrations in two plasma samples, classied by breed,
age, and body condition score at the start of the breeding program.
Variable

Level

Pubertal (%)

Breed

Friesian
Jersey
Other
12.2-14.2
14.3
14.5
14.8-15.9
3.0-4.0
4.5
5.0
5.5-7.5

45.1a
86.7b
70.0c
52.5a
59.1a
64.0b,c
65.9b,c
37.9a
66.8b
72.3b
74.3b

Age (mo)

Body condition score

ac
Within an end point, means without a common superscript differed
(P < 0.05).

This study demonstrated that synchronization of heifers

using a combination of GPG and P4 with FTAI coincident
with the nal GnRH treatment (Cosynch P4) resulted in
superior reproductive and economic performance compared with a similar program but with FTAI delayed by
approximately 24 hours or to a Double PG program.
The conception rate to AI of 57% for heifers in the
Cosynch P4 group in the current study was comparable
with the 53% reported after a previous synchrony program
with FTAI in New Zealand [6], and was superior to results
for the GPG program in Holstein heifers in the United States
when heifers detected in estrus before the nal GnRH were
inseminated early and overall conception rates were 46%
[11]. The 21-day in-calf rate of 76% for the Cosynch P4
heifers was also better than the 32-day pregnancy rate of
60% reported for dairy heifers in Florida after an 8-day
Cosynch program using P4 inserts from Days 0 to 5 [22].
The higher conception rates to AI in the Cosynch P4
group compared with heifers in the GPG P4 group in
the current study were comparable with the nding of
Lima et al. [23], in which a Cosynch P4 protocol resulted
in improved reproductive outcomes compared with an
Ovsynch P4 protocol in dairy heifers in Florida.
In the current study, heifers observed to have lost tail
paint during the 48 hours before AI had higher conception
rates to AI than those with loss of tail paint during the 48
hours after AI. The interval from the end of a synchrony
program to estrus was affected by the size of the dominant
follicle at the end of the program, with a larger dominant
follicle associated with a shorter interval to estrus [24]. It is
not clear if the shorter interval from the end of the
synchrony program to estrus was also associated with
a shorter interval to ovulation. After a synchrony program,
the interval between onset of estrus and ovulation
appeared to be reasonably consistent at 1.3 days in a range
of synchrony systems [25]. Thus it is likely that heifers in
the current study that were in estrus earlier relative to Day
0, also ovulated sooner. This might partly explain the higher
conception rates in the Cosynch P4 program compared
with the GPG P4 program. In the latter program, nearly
half of the heifers had been detected in estrus by Day 0, yet
by design were not inseminated until 24 hours later. So, for
this group of heifers, insemination might have occurred too
late for optimum fertility. The interval between estrus and
ovulation has been reported as 38.5  3.0 hours (mean 
SEM) in dairy heifers [26]. Because sperm capacitation
takes 4 to 12 hours, logically insemination should occur
12 to 18 hours before ovulation to ensure functional sperm
are present at the time of ovulation [27]. In one study,
heifers in estrus in the 24 hours preceding FTAI had higher
conception rates than those in estrus 25 to 40 hours before
FTAI (76% vs. 50%) [28]. In another study, conception rates
to FTAI were optimized when insemination occurred 6 to
22 hours before ovulation [29].
Of the heifers in the Double PG group, 65% were
detected in estrus between 2 and 6 days after the nal
treatment with PGF2a. This appeared to be lower than in
some previous reports, in which 86% were detected in
estrus over 5 days after the nal treatment [30]. This lower

888

S. McDougall et al. / Theriogenology 79 (2013) 882889

Table 4
Partial budget, in New Zealand dollars per heifer, on the use of two programs for synchronization of estrous compared with a double PGF2 program.
Treatment Group

GPG P4a
Cosynch P4b

Additional income

Additional costs

Net benet

Extra milk production

Value of AI calves

Total

Treatment costs

Management costs

Total

Mean

95% CI

26.68
43.78

14.40
23.40

41.08
67.18

40.36
40.36

1.35
1.10

41.71
41.46

0.65
25.73

21.87 to 21.58
2.9950.69

Abbreviation: CI, condence interval.

a
GnRH, PGF2 , and GnRH on Days
9,
2, and 0 of the breeding program, respectively, with insertion of an intravaginal progesterone-releasing insert
for 7 days commencing on Day
9, and xed time AI on Day 1.
b
GnRH, PGF2 , and GnRH on Days
9,
2, and 0 of the breeding program, respectively, with insertion of an intravaginal progesterone-releasing insert for
7 days commencing on Day
9, and xed-time AI on Day 0.

detection rate might have been related to lower sensitivity

of detection of estrus by the observers, a 1-day shorter
period of observation, or because only 60% of heifers in the
current study had reached puberty when the program
started.
Pubertal status had a signicant effect on reproductive
outcomes in the current study; the percentage of heifers
that were postpubertal at the start of the study decreased
with lower BCS and younger age at commencement of the
program. In a previous study, the risk of conception to rst
service was not associated with withers height, age, BCS, or
body weight, but the proportion of heifers that were
prepubertal was not dened and it is likely that most
heifers were postpubertal, hence the biophysical measures
might have been of lesser importance [31]. In the current
study, there were signicant effects of herd on all outcome
variables, likely associated with the percentage of animals
that were prepubertal in each herd. At a practical level, our
ndings demonstrated that optimal outcomes after
synchrony programs will be achieved in heifers that are
postpubertal, and as such should be 14.5 months old and
have a BCS >4.0.
In addition to improved reproductive performance, the
Cosynch P4 program had a signicant economic benet
compared with the Double PG program. One potential
benet of use of AI in heifers is a shortening of the generation interval with an associated faster rate of genetic gain.
In a previous assessment of the economic benets of articially inseminating heifers with or without synchrony, the
optimal economic decision was to select high genetic merit
heifers and undertake AI on detection of estrus [1].
However, in that analysis, the main benet modeled was
the value of the improved genetics of the calves, and it
ignored any potential benets that synchrony might have
in terms of earlier calving of the heifers with resultant
increased lactation length. The current study analyzed the
returns in terms of extra value associated with AI-sired
calves, and additional production in the rst lactation.
The economic benet of the Cosynch P4 program was
because of the 100% submission rate and the higher
conception rate that resulted in more AI-sired calves and
earlier calving, and thus a longer rst lactation length
compared with the other programs. Although the Double
PG program was less expensive, the poorer pregnancy rates
resulted in poorer economic outcomes relative to the
Cosynch P4 treatment. In populations of heifers with
fewer prepubertal animals, the Double PG system might be
more effective and less economically disadvantaged.

4.1. Conclusions
Synchrony of dairy heifers with GPG P4 and with FTAI
at the time of the nal GnRH injection (Cosynch P4)
resulted in the highest biological and economic outcomes
of the three programs tested. Pubertal status had a signicant effect on reproductive performance, irrespective of
synchrony program.
Acknowledgments
The contribution of the herd owners, staff, and the
graziers is gratefully acknowledged. The support of the
technical team of Laura Clausen, Catherine Sheehan, Aimee
Wilson, and Cathy Yanez is also gratefully acknowledged.
Progesterone RIA were undertaken by Angela Sheehan of
DairyNZ. Treatments and pregnancy testing were undertaken by the authors and by Dr Megan Clements and
Dr Geoff Plant of Anexa Animal Health.
The funding and product support provided by Bayer
Animal Health New Zealand Ltd. and by DairyNZ via an
On-farm innovation fund grant (OF811) is acknowledged.
Semen was provided by LIC, who were also helpful in
organizing inseminators for the study.
References
[1] Rendel JM, Xu ZZ, Burton LJ. Articial insemination of yearling
heifers: an economic analysis. Proc Soc Dairy Cattle Vet NZVA 1998:
25562.
[2] Anonymous. New Zealand dairy statistics 2010/11. Hamilton, NZ:
LIC and DairyNZ; 2011.
[3] Macmillan KL, Taufa VK, Day AM. Combination treatments for
synchronising oestrus in dairy heifers. Proc NZ Soc Anim Prod 1993;
53:26770.
[4] Hanlon DW, Williamson NB, Wichtel JJ, Steffert IJ, Craigie AL,
Pfeiffer DU. The effect of estradiol benzoate administration on
estrous response and synchronized pregnancy rate in dairy heifers
after treatment with exogenous progesterone. Theriogenology
1996;45:77585.
[5] Hanlon DW, Williamson NB, Wichtel JJ, Steffert IJ, Craigie AL,
Pfeiffer DU. Ovulatory responses and plasma luteinizing hormone
concentrations in dairy heifers after treatment with exogenous
progesterone and estradiol benzoate. Theriogenology 1997;47:
96375.
[6] Xu ZZ, Burton LJ. Reproductive performance of dairy heifers after
estrus synchronization and xed-time articial insemination. J Dairy
Sci 1999;82:9107.
[7] Macmillan KL, Curnow RJ, Morris GR. Oestrus synchronisation with
a prostaglandin analogue: 2. Systems in maiden heifers. NZ Vet J
1978;26:93103.
[8] Refsal KR, Seguin BE. Effect of stage of diestrus and number of
cloprostenol (ICI 80, 996) injections on intervals to estrus, LH peak
and ovulation in heifers. Theriogenology 1980;14:3748.

S. McDougall et al. / Theriogenology 79 (2013) 882889

[9] Tanabe TY, Hann RC. Synchronized estrus and subsequent conception in dairy heifers treated with prostaglandin F2a. I. Inuence of
stage of cycle at treatment. J Anim Sci 1984;58:80511.
[10] Patterson DJ, Kojima FN, Smith MF. A review of methods to
synchronize estrus in replacement beef heifers and postpartum
cows. J Anim Sci 2003;81:E16677.
[11] Schmitt EJ, Diaz PT, Drost M, Thatcher WW. Use of a gonadotropinreleasing hormone agonist or human chorionic gonadotropin for
timed insemination in cattle. J Anim Sci 1996;74:108491.
[12] Pursley JR, Wiltbank MC, Stevenson JS, Ottobre JS, Garverick HA,
Anderson LL. Pregnancy rates per articial insemination for cows
and heifers inseminated at a synchronized ovulation or synchronized estrus. J Dairy Sci 1997;80:295300.
[13] Pursley JR, Mee MO, Wiltbank MC. Synchronisation of ovulation in
dairy cows using PGF2a and GnRH. Theriogenology 1995;44:91523.
[14] Dahlen CR, Lamb GC, Zehnder CM, Miller LR, DiCostanzo A. Fixedtime insemination in peripuberal, lightweight replacement beef
heifers after estrus synchronization with PGF2alpha and GnRH.
Theriogenology 2003;59:182737.
[15] Martinez MF, Kastelic JP, Adams GP, Cook B, Olson WO, Mapletoft RJ.
The use of progestins in regimens for xed-time articial insemination in beef cattle. Theriogenology 2002;57:104959.
[16] Rivera H, Lopez H, Fricke PM. Use of intravaginal progesteronereleasing inserts in a synchronization protocol before timed AI
and for synchronizing return to estrus in Holstein heifers. J Dairy Sci
2005;88:95768.
[17] Lucy MC, Billings HJ, Butler WR, Ehnis LR, Fields MJ, Kesler DJ, et al.
Efcacy of an intravaginal progesterone insert and an injection of
PGF2alpha for synchronizing estrus and shortening the interval to
pregnancy in postpartum beef cows, peripubertal beef heifers, and
dairy heifers. J Anim Sci 2001;79:98295.
[18] Richardson AM, Hensley BA, Marple TJ, Johnson SK, Stevenson JS.
Characteristics of estrus before and after rst insemination and
fertility of heifers after synchronized estrus using GnRH, PGF2, and
progesterone. J Anim Sci 2002;80:2792800.
[19] McGowan MR, Carroll CL, Davies FJ. Fixed-time insemination of Bos
indicus heifers following the use of Syncro-Mate-B (Smb) to
synchronize estrus. Theriogenology 1992;37:1293300.
[20] Roche JR, Dillon PG, Stockdale CR, Baumgard LH, VanBaale MJ.
Relationships among international body condition scoring systems.
J Dairy Sci 2004;87:30769.

889

[21] Holm S. A simple sequentially rejective multiple test procedure.

Scand J Stat 1979;6:6570.
[22] Rabaglino MB, Risco CA, Thatcher MJ, Kim IH, Santos JEP,
Thatcher WW. Application of one injection of prostaglandin F2a in
the ve-day Co-Synch CIDR protocol for estrous synchronization and resynchronization of dairy heifers. J Dairy Sci 2010;93:
10508.
[23] Lima FS, Ayres H, Favoreto MG, Bisinotto RS, Greco LF,
Ribeiro ES, et al. Effects of gonadotropin-releasing hormone at
initiation of the 5-d timed articial insemination (AI) program
and timing of induction of ovulation relative to AI on ovarian
dynamics and fertility of dairy heifers. J Dairy Sci 2011;94:
49975004.
[24] Savio JD, Thatcher WW, Badinga L, de la Sota RL, Wolfenson D.
Regulation of dominant follicle turnover during the oestrous cycle
in cows. J Reprod Fertil 1993;97:197203.
[25] Martinez MF, Adams GP, Kastelic JP, Bergfel DR, Mapletoft RJ.
Induction of follicular wave emergence for estrus synchronization
and articial insemination in heifers. Theriogenology 2000;54:
75769.
[26] Saumande J, Humblot P. The variability in the interval between
estrus and ovulation in cattle and its determinants. Anim Reprod Sci
2005;85:17182.
[27] Vishwanath R, Nebel RL, McMillan WH, Pitt CJ, Macmillan KL.
Selected times of insemination with microencapsulated bovine
spermatozoa affect pregnancy rates of synchronized heifers. Theriogenology 1997;48:36976.
[28] Smith RD, Pomerantz AJ, Beal WE, McCann JP, Pilbeam TE, Hansel W.
Insemination of Holstein heifers at a preset time after estrous cycle
synchronization using progesterone and prostaglandin. J Anim Sci
1984;58:792800.
[29] Tenhagen BA, Kuchenbuch S, Heuwieser W. Timing of ovulation
and fertility of heifers after synchronization of estrus with
GnRH and prostaglandin F-2 alpha. Reprod Domest Anim 2005;
40:627.
[30] Kaim M, Rosenberg M, Folman Y. Management of reproduction in
dairy heifers based on the synchronisation of estrous cycles. Theriogenology 1990;34:53747.
[31] Donovan GA, Bennett FL, Springer FS. Factors associated with rst
service conception in articially inseminated nulliparous Holstein
heifers. Theriogenology 2003;60:6775.