Theriogenology 63 (2005) 1529–1548

www.journals.elsevierhealth.com/periodicals/the

Reproductive performance in anestrous dairy

cows following treatment with two protocols
and two doses of progesterone
S. McDougalla,*, C.W.R. Comptona, D.W. Hanlonb,
P.J. Davidsonb, D.J. Sullivana, A.H. Gorea, F.M. Annissa
a
Animal Health Centre, P.O. Box 21, Morrinsville, New Zealand
b
Matamata Veterinary Services, 26 Tainui St., Matamata, New Zealand
Received 8 August 2003; accepted 30 March 2004

Abstract

The objectives of this study were to evaluate the effect using two doses of progesterone (P4)
releasing devices in two different programs on reproductive performance of anestrous dairy cows.
Cows (n = 1555) not detected in estrus by 10 d before the planned start of the seasonal breeding
program and in which no CL was palpable were treated with an intravaginal P4-releasing device
(‘Single’; 1.56 g of P4) or a modified device with triple the normal P4 dose (‘Triple’; 4.7 g of P4).
The devices were in place for either 6 d (‘Short’) or 8 d (‘Long’), with 1 mg estradiol benzoate (EB)
given 24 h after device removal. The ‘Long’ program also included treatment with 2 mg EB at device
insertion. The Long program resulted in a higher first service conception rate (RR = 1.18 (95%
CI = 1.03–1.33); P = 0.02), but had no effect on the 28-d, 56-d or final pregnancy rate compared to the
Short program. There were no effects of dose of P4 on any outcome. In conclusion, the Long
compared to the Short program, but not the dose of P4, improved first service conception rates in
anestrous cows.
# 2004 Elsevier Inc. All rights reserved.

Keywords: Anestrus; Progesterone; Dairy cattle; Conception rate

* Corresponding author. Tel.: +64 7 889 5159; fax: +64 7 889 3681.
E-mail address: smcdoug@ahc.co.nz (S. McDougall).

0093-691X/$ – see front matter # 2004 Elsevier Inc. All rights reserved.
doi:10.1016/j.theriogenology.2004.03.024
1530 S. McDougall et al. / Theriogenology 63 (2005) 1529–1548

1. Introduction

Anovulatory anestrus is a reproductive problem within the New Zealand dairy

industry with 20% of cows not detected in estrus by the start of the seasonal breeding
program [1]. Anestrus is associated with lower insemination, conception and pregnancy
rates as well as a greater probability of being culled for failure to conceive [1]. The
present treatment of anestrus is 5–6 d of intravaginal progesterone (P4) treatment
followed 1–2 d later by injection of 0.5–1 mg of estradiol benzoate im (EB) [1–3] which
results in approximately 90% of cows undergoing estrus and ovulation and 45% of the
inseminated cows conceiving to the first service [1]. However, this conception rate is
lower than for cycling herd mates that have an average conception rate of 55% [4]. One
reason for this lower conception rate may be insufficient duration or concentration of P4
‘priming’ following treatment with the currently available intravaginal P4-releasing
devices.
Peak luteal P4 concentrations increase with sequential luteal phases postpartum [5].
The conception rate to insemination also increases with the number of estrous
cycles postpartum [6,7] and with time postpartum [1]. A positive correlation between
serum P4 concentration in the previous diestrous phase and the probability of conception
has been demonstrated [8–11]. The P4 concentrations in the preceding luteal phase may
influence subsequent conception rate either by altering follicle wave dynamics or by
effects on the conceptus or uterus. In cycling cows, low plasma P4 concentrations
associated with exogenous P4 treatments result in ineffective suppression of LH release,
development of abnormally large and persistent dominant follicles, high estradiol
production, premature oocyte activation, and depression of subsequent conception rates
[12–14]. In contrast, if small follicles are ovulated, the resultant conception rate is lower
and plasma P4 concentration in the subsequent luteal phase reduced [15]. Progesterone
also has a direct role in modulating uterine secretion, essential for early conceptus
development [16]. Progesterone enhances the ability of the fetus to elongate and to
produce interferon t, the fetal signal essential for pregnancy recognition, and P4 restricts
oxytocin receptor formation in the endometrium, an essential precursor for luteolysis
[17]. The oviducts from cows with a persistent dominant follicle have different
biosynthetic activity than oviducts from cows without a persistent dominant follicles;
this may also affect fertility [18]. Hence, there is evidence that P4 concentrations in the
previous luteal phase can affect follicle development and subsequent fertility and that P4
concentration in the luteal phase following insemination may also affect probability of
conceptus survival.
Plasma or serum P4 concentrations can be increased by increasing the dose of
exogenous P4 delivered. For example, increasing the dose of P4 moulded into an
intravaginal P4-releasing device or increasing the number of devices inserted results in a
curvilinear increase in plasma P4 concentrations [19,20].
Cows not detected in estrus but found to have ovulated (i.e. a corpus luteum (CL) is
detected upon palpation) have poorer reproductive performance than herdmates detected in
estrus before the start of the seasonal mating program [21]. Treatment of these animals with
various combinations of P4, EB and PG has resulted in inconstant responses compared to
no treatment [3,22,23].
 S. McDougall et al. / Theriogenology 63 (2005) 1529–1548 1531

It was hypothesized that increasing the P4 dose, or modifying the program by extending
the duration of the P4 treatment and adding EB at device insertion, would enhance the
first service conception rate in anestrus dairy cattle.

2. Materials and methods

Cows (n = 1883 from 17 dairy herds) not detected in estrus but calved >30 d at the
planned start of the seasonal breeding program were presented for veterinary examination
and assessment of body condition score (on a 1–10 scale; [24]) 10 d before the start of the
breeding program. The average interval from calving to the start of the breeding program
was 64 d (range 32–121 d).
Cows without a corpus luteum and without uterine or ovarian pathology upon rectal
palpation (n = 1555; 83% of not detected in estrus cows) were defined as anestrus and
assigned to treatment in a 2  2 factorial arrangement. The four treatments were
randomly applied with each sequential four cows presented for examination. The
treatments were either a Single or Triple P4-releasing device for a period of either 6 or 8
d. The P4-releasing device was a commercially available device (Cue-Mate Cattle
Device; Pfizer Animal Health, Auckland, New Zealand). The device consists of a
reusable nylon ‘wishbone’ onto which are threaded, two silastic, P4-impregnated ‘pods’
containing a total of 1.56 g of P4. The device was modified by cutting off the closed end
of four ‘pods’ and threading two of each of these onto each ‘arm’ of the wishbone before
the addition of two intact ‘pods’ to hold the ‘pods’ in place on the wishbone. Thus a total
of  4.7 g of P4 was incorporated into the Triple P4-releasing device. Modification of
the P4-releasing device resulted in an increase from 1.2  0.2 to 2.9  0.2 ng/mL
average (S.E.M.) plasma P4 concentrations over 8 d of insertion (P < 0.01; McDougall
et al., unpublished). The Long treatment also included 2 mg EB im at device insertion
and all cows were treated with 1 mg of EB im, 24 h after device removal. The insertion of
the P4-releaseing devices for the Short treatment occurred 2 d after the Long treatment
so that the final EB treatment occurred on the same day (Day 1) for all cows. Day 0 was
the start of the breeding program.
Those cows not detected in estrus but found to have a CL upon veterinary examination
were defined as cycling and alternately assigned at presentation to be treated either with a
Single or Triple P4-releasing device for a period of 8 d with injection of 2 mg EB im at
device insertion and 1 mg IM, 24 h after device removal.
Cows that were detected in estrus for an average 36 d (range = 23–51 among herds) after
the start of the breeding program were submitted for artificial insemination (AI). Following
this, bulls were placed with the cows in each herd for a further 63 d (range = 39–93 among
herds) so that the average herd breeding period was 99 d (range = 69–142 d). Every cow
was pregnancy tested by transrectal ultrasonography (Aloka 500 with a 5 MHz linear array
transducer; Medtel, Auckland New Zealand) approximately 10 weeks after the
commencement of the seasonal breeding program and again 6 weeks after bulls were
finally removed from the herd. The stage of gestation was estimated on the first occasion
that pregnancy was detected for each cow and from these data conception dates were
estimated. Where this estimated conception date was within 7 d of a recorded AI or
1532 S. McDougall et al. / Theriogenology 63 (2005) 1529–1548

herdowner-recorded bull breeding date, the recorded date was used as the conception date
for further calculations. If the estimated conception date was >7 d after the last recorded
breeding date, the estimated date was used for subsequent calculations.
Only one of the 17 herds did not routinely record milk production and composition
every 4–8 weeks. The production data closest to the start of the seasonal breeding program
was used for analysis. This was on average 12 d before the start of the breeding program
(range 30 d before to 42 d after start of the breeding program among herds).
Data collected for each enrolled cow included herd, breed (coded as Friesian (F) if >11/
16th Friesian, Jersey (J) if >11/16th Jersey or crossbred (XB; all others)), calving date, age
(coded as 2, 3, 4–5 and >5 years), milk volume, milk protein percentage, milk fat
percentage, date of the first herd test of the season, body condition score at the time of
initial examination, every AI and bull breeding date, culling reasons and date (if
applicable), and estimated conception date. Data were retrieved from the national data base
via the DairyWin or MindaPro programs (Livestock Improvement Corporation, Hamilton,
New Zealand) and from herdowner or veterinary practice records.
Blood samples were drawn from the tail vein of seven or eight cows randomly selected
from each group (i.e. Short Single P4, Short Triple P4, Long Single P4 and Long Triple P4)
within each of four herds on Day 13. Samples were drawn into evacuated glass tubes
containing lithium heparin anti-coagulant (Vacutainer, Becton Dickinson, Franklin Lakes,
NJ, USA), placed on ice and centrifuged at 1000  g within 4 h of collection. The plasma
was stored at 20 8C before analysis. Plasma P4 concentrations were determined using a
commercial I125 labeled RIA (Coat-A-Count; Diagnostic Products Corporation, Los
Angeles, CA, USA). Samples were analyzed within one assay and the coefficients of
variation were 19.8, 5.7 and 6.4% for quality control sera (n = 8) with mean P4
concentrations of 0.5, 3.0 and 4.2 ng/mL, respectively. The minimum detectable
concentration of the assay was 0.09 ng/mL.

2.1. Statistical analyses

The specific hypotheses being tested were: (i) that the Triple P4-releasing device would
increase first service conception rate compared to the Single P4-releasing device; and (ii)
that Long program would increase first service conception rate compared to the Short
program. Additionally, a number of other risk factors for reproductive performance were
examined including milk composition and body condition score.
A number of cows (n = 84) were lost to follow-up or excluded. Exclusion criteria
included pregnancy at initial examination (n = 1), death prior to initial pregnancy diagnosis
(n = 3), detection in estrus between initial examination and insertion of the Short P4-
releasing device (n = 18), the loss of the P4-releasing device before due day of removal (n =
53) or not being presented for EB treatment (n = 9). Cows that died or were culled between
the start of the breeding program and the first pregnancy diagnosis were only included in
the submission rate analysis.
The outcome variables examined were:

 Submission rate by Day 7 (% of cows; i.e. number of cows inseminated within 7 d of the
start of the breeding program/number of enrolled cows),
 S. McDougall et al. / Theriogenology 63 (2005) 1529–1548 1533

 Submission rate by Day 28 (% of cows; i.e. number of cows inseminated within 28 d of

the start of the breeding program/number of enrolled cows),
 Conception rate to first service (% inseminations; number of cows conceiving within 7 d
of the start of the breeding program/number of enrolled cows inseminated within 7 d of
start of the breeding program),
 Pregnancy rate by Day 28 (% of cows; number of cows conceiving within 28 d of the
start of the breeding program/number of enrolled cows),
 Pregnancy rate by Day 56 (% of cows; number of cows conceiving within 56 d of the
start of the breeding program/number of enrolled cows),
 Final pregnancy rate (% of cows; number of cows conceiving/number of enrolled cows),
and the
 Interval from the start of the breeding program to conception (days; interval from start of
the breeding program to conception).

The independent (explanatory) variables included:

 Dose of P4 (Categorical; coded as ‘Single’ P4 (reference category) and ‘Triple’ P4),

 ‘‘Duration’’ of P4 (Categorical; coded as ‘Short’ (reference category) and ‘Long’),
 Postpartum interval (PPI) (Continuous as days),
 Postpartum interval (PPI; days) (Categorical ordinal, coded as >76 d = Early, 64–76 d =
Early–mid, 50–63 d = Mid–Late, <50 d = Late),
 Postpartum interval (PPI; d) (Categorical ordinal, coded as 50 d = Early, <50 d =
Late),
 Milk fat % at first herd test (Continuous as a %),
 Milk volume at first herd test (Continuous as L/cow/d),
 Protein % at first herd test (Continuous as a %),
 Protein % at first herd test (Categorical ordinal, coded as >3.73% = High, 3.52–3.73% =
Med–High, 3.33–3.51% = Med–Low and <3.33% = Low),
 Protein % at first herd test (Categorical ordinal, coded as 3.76% = High, <3.76% =
Low),
 Fat to protein ratio (Continuous),
 Fat to protein ratio (Categorical ordinal, coded as <1.14 = Low, 1.14–1.22 = Med–Low,
1.23–1.34 = Med–High, >1.34 = High),
 Body condition score (Categorical ordinal, coded as 4 = High, <4 = Low),
 Breed code (Categorical, coded as Friesian = 1, Crossbred = 2, Jersey = 3),
 Age code (years; Categorical ordinal, coded as 2 = 2, 3 = 3, 4–5 = 4, >5 = 5).

The lowest value within a categorical variable is the reference category unless stated
otherwise.
For anestrus cows, logistic regression was performed for each outcome variable (except
final pregnancy rate and the start of the breeding program to conception interval). The
independent (explanatory) variables included dose (i.e. Single P4 versus Triple P4),
program (i.e. Short versus Long), herd, age code, breed, body condition score at the time of
enrolment, milk fat to protein ratio, milk protein percentage and postpartum interval.
1534 S. McDougall et al. / Theriogenology 63 (2005) 1529–1548

Initially, all explanatory variables were screened by univariate analyses (x2 or logistic
regression), and then variables associated (P < 0.25) were offered to forward and reverse
stepwise logistic regression models using likelihood ratio for variable inclusion/exclusion.
The main effects of dose and program were always included in the models, even if not
significant. The dose by program interaction was explicitly tested for within each analysis.
It was only significant in the Day 56 pregnancy rate model. The validity of the models was
assessed using the Hosmer–Lemeshow test and counting the list of outliers (i.e. values >2
standard deviations from expected) to ensure that <5% of values were outliers. The results
from the logistic regression analyses were converted from odds ratios to relative risks (RR)
using the methodology of Zhang and Kai [25] as the adjusted odds ratio derived from the
logistic regression is not a good approximation of relative risk when the outcome of interest
(pregnancy in this study) is not rare. A relative risk of >1 and <1 indicates that the risk
factor increases or decreases the risk of conception or pregnancy, respectively.
Every enrolled cow in two herds finally conceived (Herds 1 and 6) resulting in unstable
coefficient estimates with attempts to model the final pregnancy rate of anestrus cows using
logistic regression. Consequently, the final pregnancy rate was analyzed using the Mantel–
Haenszel technique independently for each of the main effects listed above stratified by
herd. This was done as herd was significant in an initial x2 analysis and regarded as a
potentially confounding factor.
The median interval from the start of the breeding program to conception for each
treatment was estimated using Kaplan–Meir survival analysis. The probability of
conception was formally tested using forward stepwise Cox’s proportional hazards survival
analysis with the same independent variables as above. The data are presented as median
days, standard error and 95% confidence intervals (95% CI) (from the Kaplan–Meir
analysis) and the hazards ratio (HR) with P-values derived from the final Cox’s model. The
validity of the Cox’s model was evaluated by visualizing the Log–Log hazard versus time
plots to ensure that the hazards assumption was not violated. The partial residuals and the
coefficients with and without each case (i.e. the DfBeta’s) were plotted and no violations of
proportional hazards assumptions or extreme outliers were detected.
The numbers of not detected in estrus but CL-positive cows enrolled was low. Attempts
to model reproductive outcomes using both logistic regression and Mantel–Haenszel
techniques resulted in unstable models and a failure in convergence in logistic regression
models. Data for these cows were presented as raw data and relative risks from the x2
analysis with dose as the explanatory variable.
The Day 13 plasma P4 concentration was analyzed in a general linear model with the
main effects of dose (Single P4 versus Triple P4) and program (Short versus Long) as fixed
effects, with herd, age code (2, 3, 4–5, and >5 years) and conception to first service within
7 d of the start of the breeding program initially included as random effects and with milk
fat percentage, milk protein percentage and fat to protein ratio initially included as
covariates. Initially, univariate (i.e. one-way ANOVA and linear regression) analysis was
used to analyze the relationship between plasma P4 and the effect in question. Those that
were found to be associated (i.e. P < 0.2) and those thought important in the design (i.e.
herd, P4 dose and program and conception to first service) were initially included in the
model. A manual, backward model building process was used and effects not significant
(i.e. P > 0.1) were removed. All first order interactions of remaining effects were tested and
 S. McDougall et al. / Theriogenology 63 (2005) 1529–1548 1535

removed if not significant (P > 0.1). Plasma P4 data are presented as least square means
and the standard errors of the difference (S.E.D.). The P4 concentration was arbitrarily
defined as low and high for concentrations of <1 ng/mL and >1 ng/mL, respectively. The
proportion of cows with low P4 for each dose (Single P4 versus Triple P4) and program
(Short versus Long) of treatment as well as conceiving or not conceiving to first
insemination was examined using x2.
The program SPSS (v11.5; SPSS Inc., Chicago, IL, USA) was used for data analysis.

3. Results

A total of 1883 cows were initially presented as not detected in estrus (Table 1). The
level of prevalence of cows not detected in estrus averaged 31.2% (range 9.1–52.5% among
herds; Table 1).

3.1. Reproductive performance of anestrus cows

3.1.1. 7-d submission rate

There was no effect of program or dose of P4 on the 7-d submission rate (Tables 2 and
3). The 7-d submission rate varied among herds (average = 90.4%, range = 74.0–98.7%, P
< 0.01) and was lower in cows with high (4) body condition score compared to cows in
low (<4) body score (Table 3).

Table 1
Descriptive data for enrolled herds
Herd code Total cows (n) Enrolled cows (n) Not detected in estrus (%)a Anestrus (%)b
1 340 31 9.1 93.5
2 372 80 21.5 75.0
3 467 122 26.1 86.1
4 523 157 30.0 80.3
5 370 103 27.8 95.1
6 179 39 21.8 69.2
7 253 59 23.3 71.2
8 431 182 42.2 84.6
9 360 88 24.4 94.3
10 160 84 52.5 82.1
11 142 67 47.2 80.6
12 195 82 42.1 92.7
13 469 167 35.6 90.4
14 386 101 26.2 79.2
15 638 297 46.6 67.3
16 343 105 30.6 86.7
17 510 119 23.3 92.4
a
Percentage of all cows in the herd not detected in estrus.
b
Percentage of not detected in oestrus cows diagnosed as anestrus (i.e. no CL was detected upon palpation of
the ovaries).
1536 S. McDougall et al. / Theriogenology 63 (2005) 1529–1548

Table 2
Submission (Sub), conception (Con) and pregnancy (Preg) rates (number and %) for anestrus cows treated with a
Single or Triple P4-releasing device (Dose) for either a ‘Short’ or ‘Long’ duration (Program)
Outcome Dose Program
Single Triple Short Long
No. % No. % No. % No. %
Sub D7 707/784 90.2 700/771 90.8 699/777 90.0 708/778 91.0
Con S1 273/707 38.6 277/700 39.6 248/699 35.5 302/708 42.7
Sub D28 757/784 96.6 742/771 96.2 743/777 95.6 756/778 97.2
Preg D28 467/784 59.6 450/771 58.4 447/777 57.5 470/778 60.4
Preg D56 605/753 80.3 589/726 81.1 590/739 79.8 604/740 81.6
Preg Final 615/667 92.2 598/648 92.3 605/654 92.5 608/661 92.0
PSB-cona 21 20–22 21 20–22 21 20–22 20 19–21
a
Median interval (and 95% confidence intervals) of the days from the planned start of the seasonal breeding
program (PSB) to conception from Kaplan–Meier analysis.

Table 3
Relative risk (RR), 95% confidence intervals (95% CI, lower and upper CI = LCL and UCL, respectively) and P-
value for the final model of 7-d submission rate of anestrus cows treated with a Single or Triple P4-releasing device
(Dose) for either a ‘Short’ or ‘Long’ duration (Program)
RR 95% CI P
LCL UCL
P4 dosea 1.01 0.97 1.03 0.66
Programb 1.01 0.98 1.04 0.44
BCS 4c 0.92 0.83 0.99 0.001
a
Reference category: Single P4 device.
b
Reference category: Short (6D + EB) program.
c
Reference category: BCS <4.

Table 4
Relative risk (RR), 95% confidence intervals (95% CI, lower and upper CI = LCL and UCL, respectively) and P-
values for the final model of first service conception rate for anestrus cows treated with a Single or Triple P4-
releasing device (Dose) for either a ‘Short’ or ‘Long’ duration (Program)
Category RR 95% CI P
LCL UCL
P4 dosea 1.00 0.85 1.17 0.95
Programb 1.18 1.03 1.33 0.023
Age group (years)c 2 1.51 1.23 1.82 0.000
3 1.37 1.08 1.69 0.006
4+5 1.52 1.22 1.83 0.000
Fat to protein ratiod Low 1.16 0.92 1.42 0.204
Med–low 0.99 0.77 1.23 0.902
Med–high 1.25 1.01 1.49 0.039
a
Reference category: Single P4 device.
b
Reference category: Short (6D + EB) program.
c
Reference category: age >5 years.
d
Reference category: milk fat to protein ratio ‘high’ (i.e. >1.34).
 S. McDougall et al. / Theriogenology 63 (2005) 1529–1548 1537

Table 5
Relative risk (RR), 95% confidence intervals (95% CI, lower and upper CI = LCL and UCL, respectively) and P-
values for the final model of 28-d submission rate for anestrus cows treated with a Single or Triple P4-releasing
device (Dose) for either a ‘Short’ or ‘Long’ duration (Program)
Category RR 95% CI P
LCL UCL
P4 dosea 1.011 0.997 1.018 0.80
Programb 0.997 0.969 1.015 0.11
Age group (years)c 3 1.045 1.016 1.056 0.010
4+5 1.025 0.987 1.044 0.15
>5 1.034 1.005 1.048 0.027
a
Reference category: Single P4 device.
b
Reference category: Short (6D + EB) program.
c
Reference category: age 2 years.

3.1.2. Conception rate to first service

The first service conception rate was higher following the Long than Short program
(P = 0.02) but was not affected by the dose of P4 (P = 0.95; Tables 2 and 4). First service
conception rate tended to vary among herds (average = 38.8%, range = 26.3–55.4%,
P = 0.08), was higher in 2, 3 and 4 to 5-year-old cows relative to cows >5 years (P < 0.001;
Table 4). Medium–high fat to protein ratios resulted in higher first service conception rates
than high fat to protein ratios (P = 0.04; Table 4).

3.1.3. 28-d submission rate

There was no effect of program (P = 0.11) or dose (P = 0.80; Tables 2 and 5) of P4 on the
28-d submission rate. The 28-d submission rate varied among herds (average = 95.7%,
range = 81.5–100.0%; P < 0.01) and was higher in 3-year-old and >5-year-old cows
compared to 2-year-old cows (Table 5).

3.1.4. 28-d pregnancy rate

The 28-d pregnancy rate was unaffected by either the program (P = 0.31) or dose
(P = 0.60) of P4 (Tables 2 and 6). Herds varied in 28-d pregnancy rate (average = 59.0%,

Table 6
Relative risk (RR), 95% confidence intervals (95% CI, lower and upper CI = LCL and UCL, respectively) and P-
values for the final model of 28-d pregnancy rate for anestrus cows treated with a Single or Triple P4-releasing
device (Dose) for either a ‘Short’ or ‘Long’ duration (Program)
RR 95% CI P
LCL UCL
P4 dosea 1.025 0.931 1.115 0.60
Programb 1.049 0.954 1.139 0.31
BCSc 0.828 0.719 0.938 0.002
a
Reference category: Single P4 device.
b
Reference category: Short (6D + EB) program.
c
Reference category: High (4) body condition score.
1538 S. McDougall et al. / Theriogenology 63 (2005) 1529–1548

Table 7
Relative risk (RR), 95% confidence intervals (95% CI, lower and upper CI = LCL and UCL, respectively) and
P-values for 56-d pregnancy rate for anestrus cows treated with a Single or Triple P4-releasing device (Dose) for
either a ‘Short’ or ‘Long’ duration (Program)
Category RR 95% CI P
LCL UCL
P4 dosea 0.986 0.926 1.038 0.63
Programb 1.010 0.949 1.060 0.74
Breedc Friesian 1.128 0.964 1.213 0.107
Crossbred 1.160 1.024 1.228 0.027
Calving to start of breedingd <50 d 0.892 0.791 0.976 0.008
50–63 d 0.961 0.870 1.031 0.305
64–76 d 1.005 0.925 1.064 0.899
BCSe 1.086 1.025 1.133 0.008
Milk protein %f 1.084 1.016 1.135 0.019
a
Reference category: Single P4 device.
b
Reference category: Short (6D + EB) program.
c
Reference category: Jersey.
d
Rerence category: calved >77 d at the start of the breeding program.
e
Reference category: Low (<4) body condition score.
f
Reference category: Low (<3.76) % milk protein.

range = 35.2–86.2%; P < 0.001). The 28-d pregnancy rate increased with increasing
interval from calving to start of the breeding program (P < 0.001) and with the protein
percentage of milk at herd test (P < 0.01), but decreased in cows with low versus high body
condition score (P < 0.01; Table 6).

3.1.5. 56-d pregnancy rate

The 56-d pregnancy rate was unaffected by either the program (P = 0.74) or dose (P =
0.63; Tables 2 and 7) of P4. However, there was a dose by program interaction (P = 0.04),
whereby the Triple P4-Long group had a higher 56-d pregnancy rate than the Single P4-
Long group (P = 0.06), while there was no difference (P = 0.2) between the Single or Triple
P4-Short groups (81.7, 77.9, 79.0, and 84.3% 56-d pregnancy rates for Single P4-Short,
Triple P4-Short, Single P4-Long, Triple P4-Long groups, respectively). Herds varied in 56-
d pregnancy rate (average = 79.4%, range = 68.5–96.6%; P = 0.01). The 56-d pregnancy
rate was lower in cows with short (<50 d) compared with long (>76 d) intervals between
calving and the start of the breeding program (Table 7). Crossbred cows had a higher 56-d
pregnancy rate than Jersey cows (P = 0.03; Table 7). Cows with a high body condition score
had a higher 56-d pregnancy rate than cows in low body condition score (P = 0.01; Table 7).
Cows with high milk protein percentage at herd test had a higher 56-d pregnancy rate than
cows with low milk protein percentage (P = 0.02; Table 7).

3.1.6. Final pregnancy rate

The final pregnancy rate was unaffected by either the program (P = 0.78) or dose (P =
0.89; Tables 2 and 8) of P4. Final pregnancy rate varied among herds (average = 91.9%,
range = 83.7–100%; P < 0.05), was higher in cows with high milk protein percentage
 S. McDougall et al. / Theriogenology 63 (2005) 1529–1548 1539

Table 8
Relative risk (RR), 95% confidence intervals (95% CI, lower and upper CI = LCL and UCL, respectively) and
P-values for final model of final pregnancy rate for anestrus cows treated with a Single or Triple P4-releasing
device (Dose) for either a ‘Short’ or ‘Long’ duration (Program)
RR 95% CI P
LCL UCL
P4 dosea 1.002 0.966 1.028 0.89
Programb 0.995 0.957 1.022 0.78
Milk protein (%)c 1.050 1.010 1.068 0.021
Calving to start of breedingd 0.955 0.896 0.997 0.031
a
Reference category: Single P4 device.
b
Reference category: Short (6D + EB) program.
c
Reference category: Low (<3.76) % milk protein.
d
Reference category: calved 51 d at the start of the breeding program.

compared to low milk protein percentage (P = 0.02; Table 8) and was lower in late calving
cows (P = 0.03; Table 8).

3.1.7. Probability of conception with time (survival analysis)

The interval from the start of the seasonal breeding program to conception was
unaffected by the program (21  0 versus 20  0 median d (S.E.) from the planned start
of the seasonal breeding program to conception for Short versus Long program,
respectively; P = 0.56). Dose did not effect the interval from the start of the seasonal
breeding program to conception (21  0 versus 21  0 median d (S.E.) from planned
start of the seasonal breeding program to conception for Single versus Triple P4, P = 0.77).
The interval from the start of the seasonal breeding program to conception varied amongst
herds (P < 0.001) and was longer in cows with a short (<51 d) interval from calving to the
start of the seasonal breeding program late calving cows (Fig. 1). Cows with high milk

Table 9
Submission (Sub), conception (Con) and pregnancy (Preg) rates (number and %) for cows not detected in estrus by
7 d before the planned start of the seasonal breeding programme which were found to have a corpus luteum upon
transrectal ovarian palpation
Outcome P4 dose RRa 95% CI P
Single Triple
n % n %
Sub D7 141/162 87.0 145/166 87.3 1.00 0.92–1.08 0.93
Con S1 53/141 37.6 57/145 39.3 0.96 0.71–1.28 0.76
Sub D28 158/162 97.5 157/166 94.6 1.03 0.99–1.08 0.17
Preg D28 101/162 62.3 101/166 60.8 1.02 0.86–1.21 0.78
Preg D56 126/162 77.8 135/166 81.3 0.96 0.86–1.07 0.43
Preg Final 137/146 93.8 136/146 93.2 1.01 0.95–1.07 0.81
Cows were treated with either a Single or Triple P4-releasing device (P4 Dose).
a
Relative risk and 95% confidence interval around RR estimate.
1540 S. McDougall et al. / Theriogenology 63 (2005) 1529–1548

Fig. 1. (a) and (b) Median (and upper 95% confidence) interval from the planned start of the seasonal breeding
program to conception by quartiles of the interval from calving to the start of the breeding program ((a) upper
panel); and by quartiles of the milk protein percentage at first herd test ((b) lower panel). Bars with different
superscripts differ at P < 0.05.

protein percentage had a shorter interval from the start of the seasonal breeding program to
conception (Fig. 1).

3.2. Reproductive performance of not detected in estrus but CL-positive cows

A total of 328 (17.4%) of presented cows were detected with a CL upon examination.
The average herd prevalence of these cows was 16.4% but there was a wide range among
herds (4.9–32.7%). A total of 162 cows were treated with the Single P4 and 166 with the
 S. McDougall et al. / Theriogenology 63 (2005) 1529–1548 1541

Fig. 2. Frequency histograms of the plasma P4 concentrations (ng/mL) 13 d after the start of the seasonal breeding
program for anestrous cows treated with a Short or Long program and using a Single or Triple P4-releasing device.
There was no significant difference between the dose of P4 or the program.

Triple P4 dose. There were no significant differences between the Single and the Triple P4
dose for any of the measured reproductive outcomes (Table 9; all P > 0.2).

3.3. Day 13 plasma P4 concentrations

The average plasma P4 concentration was 4.3 (S.E. = 0.2) ng/mL (Fig. 2) and the P4
values were normally distributed. The P4 concentration was not affected by either P4 dose
(4.69 ng/mL versus 4.18 ng/mL (S.E.D. = 0.34) for Single versus Triple P4, P = 0.14) or
program (4.43 ng/mL versus 4.45 ng/mL (S.E.D. = 0.34) for Short versus Long, P = 0.95).
1542 S. McDougall et al. / Theriogenology 63 (2005) 1529–1548

Fig. 3. Milk protein percentage compared to plasma P4 concentration (ng/mL) 13 d after the start of the seasonal
breeding program for anestrous cows treated with a Short or Long program and using a Single or Triple
P4-releasing device. The lines are linear regression lines for each herd.

Herds varied in the mean P4 concentration (P < 0.01), P4 concentration was positively
associated with milk protein percentage at first herd test (P = 0.003; Fig. 3; univariate
R2 = 0.127). The P4 concentration was higher in cows in high body condition score than
in lower body condition score (P = 0.05; Fig. 4). Cows conceiving had a higher
P4 concentration than those not conceiving to the first insemination (5.2 ng/mL versus

Fig. 4. The estimated marginal mean (and standard errors) plasma P4 concentrations for cows in various body
condition scores at 13 d after the start of the seasonal breeding program for anestrous cows treated with a Short or
Long program and using a Single or Triple P4-releasing device. Bars with different superscripts differ at P < 0.05.
 S. McDougall et al. / Theriogenology 63 (2005) 1529–1548 1543

4.2 ng/mL (S.E.D. = 0.4) P4 for cows conceiving and not conceiving to first insemination,
respectively, P = 0.04). There was no interaction of dose or program with the conception to
first service (both P = 0.2). The final model adjusted R2 was 0.353.
More of the cows not conceiving had a low P4 concentration at Day 13 (i.e. <1 ng/mL)
than those conceiving (12/78 (15.4%) versus 1/41 (2.4%); P = 0.03). There were no
differences among dose (6/57 (9.5%) versus 9/54 (14.3%) of Single versus Triple P4,
respectively, P = 0.41) or program (8/62 (12.9%) versus 7/64 (10.9%) Short versus Long
program, respectively, P = 0.73) in the proportion of cows with low P4 concentrations.

4. Discussion

The original hypothesis that a program with a longer duration of P4 treatment and
additional EB at device insertion would increase the first service conception rate was
accepted. However, the dose of P4 (i.e. Single versus Triple P4) did not effect the first
service conception rate. Additionally, neither the dose nor the program affected any of the
other outcome variables studied either in anestrus or cycling cows.
One explanation of the enhanced conception rate with the Long compared to the Short
program may be related to follicle wave dynamics. The Long, but not Short program
included EB at device insertion. Treatment with EB or GnRH concurrent with insertion
of a P4-releasing device results in atresia or ovulation of the dominant follicle and
emergence of a new follicle wave in both anestrus and cycling cows [26–29]. Removal of
a dominant follicle present at the commencement of treatment prevents ovulation of a
follicle that has had an extended period of dominance. Extended periods of dominance
are associated with depressed conception rates [13] related to high estradiol levels within
the follicle and structural changes to the oocyte [30]. Treatment with either EB or GnRH
at the time of insertion of a P4 device for 7 d or 8 d resulted in an increase in conception
rate compared to no EB or GnRH [31,32]. Conversely, an alternative hypothesis would
be that the average size of the ovulated follicle will be smaller following the Short than
the Long program. Ovulation of small follicles has been associated with smaller CL and
a reduced conception rate in cycling cows [15]. However, detailed studies in anestrus
cows have suggested that ‘persistence’ of the dominant follicle does not occur following
treatment with exogenous P4 in anestrus cows as it does in cycling cows [33]. Thus, the
possibility that the increased conception rates seen are related solely to the duration of
the P4 treatment, not mediated by changes in follicle dynamics, cannot be ruled out on
the basis of the current study. The direct effect of different durations of P4 treatment in
the absence of EB at device insertion needs to be undertaken to test this hypothesis
directly.
Previous descriptive studies have demonstrated a positive correlation between serum P4
concentration in the preceding diestrous phase and the probability of conception [8–11].
No relationship was found between the dose of exogenous P4 preceding insemination and
the conception rate in the present study. This may be due to an insufficient dose of P4
delivered or a less than optimal plasma P4 profile over time. The Triple P4-releasing device
used in the current study did increase plasma P4 concentrations during the period of
treatment, but concentrations were still below those seen in normal cycling cows in the
1544 S. McDougall et al. / Theriogenology 63 (2005) 1529–1548

luteal phase (McDougall et al.; unpublished). The temporal patterns of concentrations of

P4 following exogenous P4 treatment do not mimic those of the natural luteal phase.
Manipulation of the P4 profile by use of multiple PG treatments or PG treatment followed
by insertion of two intravaginal P4-releasing devices resulted in different P4 profiles and
different follicular wave patterns between treatments [34]. This suggests that the shape of
the P4 profile as well as the peak concentration of P4 may be important in follicle wave
control.
The previously reported positive relationship between preceding luteal phase P4
concentration and conception rate may not be a direct causal relationship. If the observed
relationship were due to some unrecorded underlying factor that was positively correlated
both with serum P4 concentration and conception rate, but that had effects not mediated via
P4, ‘confounding’ may have occurred in the previous studies. Hence P4 may be acting as a
‘proxy variable’ for other factors associated with conception rate. For example, LH pulse
frequency is influenced by nutritional status [35,36] and P4 concentration is correlated with
LH pulse frequency via LH-driven P4 release from the small luteal cells [37]. Progesterone
concentration is also correlated with weight gain [10] and reduced in underfed animals
[38]. Progesterone production may also be mediated via GH and/or insulin-like growth
factor because receptors for these have been found in large luteal cells [39,40] and GH
increases secretion of P4 from luteal cells [41]. However, GH, insulin-like growth factor,
leptin and other metabolic regulators may also directly effect conception rate and fetal
survival independent of P4 [42].
The relationship between preceding P4 and conception rate also appears to be
influenced by whether the P4 is of endogenous or exogenous origin. Where cows were
treated twice with PG 14 d apart, there was a positive relationship between subsequent
conception rate and the P4 concentration 3 d preceding the second PG treatment [11].
However, where the same PG regime was used but with the addition of an intravaginal
P4-releasing device 8 d after the initial PG treatment for 7 d, the relationship was
reversed [11].
One of the potential mechanisms by which the P4 concentration in the preceding luteal
phase could influence subsequent conception rate may be via enhancement of subsequent
luteal function and hence enhanced embryo growth and survival. Cows conceiving had
higher average P4 concentrations on Day 13 and were less likely to have low (i.e. <1 ng/
mL) P4 concentrations than those not conceiving to first insemination. However, neither
the dose nor the program length affected the plasma P4 concentration or the probability of
having a low P4 concentration on Day 13 in the subsequent luteal cycle. This is despite the
fact that the first service conception rate was higher in anestrus cows treated with the Long
versus the Short program before insemination. It appears that the positive effect of the Long
program may have been via pathways that were independent of P4 concentrations in the
subsequent luteal phase. The dose of P4 and program length were not related to P4
concentration even in the initial univariate models. Inclusion of the other effects (e.g. herd,
BCS, and protein percentage) did not lead to colinearity and hence mask some effect of
treatment. Additionally, studies which have increased serum/plasma P4 concentrations
post insemination, either by providing additional exogenous P4 directly in anestrus cows
[43] or indirectly by inducing a second CL by gonadotropin treatment in cycling cows,
have also failed to enhance conception or pregnancy rates in some [44,45] but not all
 S. McDougall et al. / Theriogenology 63 (2005) 1529–1548 1545

studies [46]. It appears that factors other than P4 concentration either before or after
insemination of P4-treated anestrus cows are required to explain the lower conception rates
commonly found in anestrus compared to cycling cows.
A number of explanatory variables other than P4 treatment were found to influence
the reproductive performance of P4-treated anestrus cows; these included herd, age,
body condition score before the planned start of the breeding program, the interval
between calving and the start of the breeding program, and milk composition. Herds
differed for all reproductive indices examined. Differences in farm-level management
including nutritional management, estrus detection and sire and cow genetics may all
impact on farm-level performance. Body condition score was negatively associated with
the 7-d submission rate, but positively associated with the 28-d, 56-d and final pregnancy
rate. That the 7-d submission rate was lower in better conditioned cows is not clear and
requires further investigation. Previous studies have demonstrated a positive relationship
between body condition around insemination and reproductive performance [47,48] and
between the interval from calving to the commencement of the breeding program and
reproductive performance [1,3]. Relationships between milk composition and
reproductive performance have also previously been demonstrated. Elevated milk fat
to protein (i.e. >1.5) ratios have been associated with reduced first service conception
rates, increased numbers of inseminations per conception and increased calving to
conception intervals [49]. In the current study, cows with high fat to protein ratios (i.e.
>1.34) had depressed first service conception rates. Positive relationships have been
demonstrated between milk protein percentage and submission and pregnancy rates in
studies in Australia [50], Ireland [51] and New Zealand [23]. In common with the
previous New Zealand study, the milk protein percentage was correlated with the 28-d
and 56-d pregnancy rate, but not the submission rates or first service conception rates.
The mechanism for the relationship between milk protein percentage and fertility is
unclear but appears to operate via pathways independent of expression of estrus. A novel
finding of the current study is that the luteal (Day 13) phase P4 concentration was
positively associated with both milk protein percentage and with body condition score.
The association between luteal phase P4 concentration, body condition score and milk
composition requires further investigation.
In conclusion, increasing the dose of P4 used to treat anestrus and not detected in estrus
but CL-positive cows did not enhance any measure of reproductive performance. However,
a program with a longer duration of progesterone treatment and with addition of EB at
P4-releasing device insertion significantly improved the first service conception rate.
The increase in conception rate was independent of the dose of P4 and of the concentration
of P4 in the subsequent luteal phase.

Acknowledgement

The support of the herd owners and staff in completing this project is gratefully
acknowledged. Pfizer Animal Health (NZ) LTD kindly provided the P4-releasing devices
as well as providing financial support for the project. T. O’Donnell (Dexcel, Hamilton,
New Zealand) kindly performed the P4 RIA assays.
1546 S. McDougall et al. / Theriogenology 63 (2005) 1529–1548

References

[1] Rhodes FM, McDougall S, Burke CR, Verkerk GA, Macmillan KL. Treatment of cows with an extended
postpartum anestrous interval. J Dairy Sci 2003;86:1876–94.
[2] McDougall S, Burke CR, Macmillan KL, Williamson NB. The effect of pretreatment with progesterone on
the oestrous response to oestradiol-17b benzoate in the post-partum dairy cow. NZ Soc Anim Prod 1992;
52:157–60.
[3] McDougall S. Reproductive performance of anovulatory anoestrus postpartum dairy cows following
treatment with two progesterone and oestradiol benzoate-based protocols, with or without resynchrony.
NZ Vet J 2001;49:187–94.
[4] Xu ZZ, Burton LJ, Macmillan KL. Reproductive performance of lactating dairy cows following oestrous
synchronisation with progesterone, oestradiol and prostaglandin. NZ Vet J 1996;44:99–104.
[5] Jochle W. The individual post parturient cow: theoretical aspects of the resumption of reproductive functions.
Proc Soc Dairy Cattle Vets NZVA 1994;11:177–90.
[6] Thatcher WW, Wilcox CJ. Postpartum estrus as an indicator of reproductive status in the dairy cow. J Dairy
Sci 1973;56:608–10.
[7] Rhodes FM, Clark BA, McDougall S, Macmillan KL. Insemination at the second of two induced oestrous
periods in anoestrous dairy cows increases conception rates to first service. NZ Vet J 1999;47:39–43.
[8] Folman Y, Rosenberg M, Herz Z, Davidson M. The relationship between plasma progesterone concentration
and conception in post-partum dairy cows maintained on two levels of nutrition. J Reprod Fertil 1973;
34:267–78.
[9] Holness DH, Sprowson GW, Sherward C, Geel G. Studies on plasma progesterone concentrations and
fertility in Friesland dairy cows during the post-partum period. J Agric Sci Camb 1981;97:649–55.
[10] Fonseca FA, Britt JH, McDaniel BT, Wilk JC, Rakes AH. Reproductive traits of Holsteins and Jerseys.
Effects of age, milk yield, and clinical abnormalities on involution of cervix and uterus, ovulation, estrous
cycles, detection of estrus, conception rates and days open. J Dairy Sci 1983;66:1128–47.
[11] Folman Y, Kaim M, Herz Z, Rosenberg M. Comparison of methods for the synchronization of estrous cycles
in dairy cows. 2. Effects of progesterone and parity on conception. J Dairy Sci 1990;73:2817–25.
[12] Sirois J, Fortune JE. Lengthening the bovine estrous cycle with low levels of exogenous progesterone: a
model for studying ovarian follicular dominance. Endocrinology 1990;127:916–25.
[13] Mihm M, Baguisi A, Boland MP, Roche JF. Association between the duration of dominance of the ovulatory
follicle and pregnancy rate in beef heifers. J Reprod Fertil 1994;102:123–30.
[14] Smith MW, Stevenson JS. Fate of the dominant follicle, embryonal survival, and pregnancy rates in dairy
cattle treated with prostaglandin F2 alpha and progestins in the absence or presence of a functional corpus
luteum. J Anim Sci 1995;73:3743–51.
[15] Vasconcelos JLM, Sartori R, Oliveira HN, Guenther JG, Wiltbank MC. Reduction in size of the ovulatory
follicle reduces subsequent luteal size and pregnancy rate. Theriogenology 2001;56:307–14.
[16] Geisert RD, Morgan GL, Short EC, Zavy MT. Endocrine events associated with endometrial function and
conceptus development in cattle. Reprod Fertil Dev 1992;4:301–5.
[17] Mann GE, Lamming GE, Robinson RS, Wathes DC. The regulation of interferon-t production and uterine
receptors during early pregnancy. J Reprod Fertil Suppl 1999;54:317–28.
[18] Binelli M, Hampton J, Buhi WC, Thatcher WW. Persistent dominant follicle alters pattern of oviductal
secretory proteins from cows at estrus. Biol Reprod 1999;61:137–44.
[19] Rathbone MJ, Bunt CR, Ogle CR, Burggraaf S, Macmillan KL, Burke CR, et al. Reengineering of a
commercially available bovine intravaginal insert (CIDR insert) containing progesterone. J Cont Rel 2002;
85:105–15.
[20] Macmillan KL, Taufa VK, Barnes DR, Day AM. Plasma progesterone concentrations in heifers and cows
treated with a new intravaginal device. Anim Reprod Sci 1991;26:25–40.
[21] McDougall S, Rhodes FM. Detection of a corpus luteum in apparently anoestrous cows by manual palpation,
transrectal ultrasonography and plasma progesterone concentrations. NZ Vet J 1999;47:47–52.
[22] Rhodes FM, McDougall S, Verkerk GA. Reproductive performance of dairy cows not detected in oestrus but
with a detectable corpus luteum, in response to treatment with progesterone, oestradiol benzoate and
prostaglandin F2a. NZ Vet J 2001;49:13–7.
 S. McDougall et al. / Theriogenology 63 (2005) 1529–1548 1547

[23] McDougall S. Resynchrony of previously anoestrous cows and treatment of cows not detected in oestrus, but
with a palpable corpus luteum with prostaglandin F2a. NZ Vet J 2003;51:117–24.
[24] Macdonald KA, Macmillan KL. Condition score and liveweight in Jersey and Friesian cows. Ruakura
Farmers Conference, Hamilton, New Zealand 1993; 45:47–50.
[25] Zhang J, Kai FY. What’s the relative risk? A method of correcting the odds ratio in cohort studies of common
outcomes JAMA 1998;280:1690–1.
[26] Crowe MA, Goulding D, Baguisi A, Boland MP, Roche JF. Induced ovulation of the first postpartum
dominant follicle in beef suckler cows using a GnRH analogue. J Reprod Fertil 1993;99:551–5.
[27] Bo GA, Adams GP, Pierson RA, Mapletoft RJ. Exogenous control of emergence of follicle waves in cattle.
Theriogenology 1995;43:31–40.
[28] McDougall S, Williamson NB, Macmillan KL. GnRH induces ovulation of a dominant follicle in
primiparous dairy cows undergoing anovulatory follicle turnover. Anim Reprod Sci 1995;39:205–14.
[29] Roche JF, Austin EJ, Ryan M, O’Rourke M, Mihm M, Diskin MG. Regulation of follicle waves to maximize
fertility in cattle. J Reprod Fertil Suppl 1999;54:61–71.
[30] Mihm M, Curran N, Hytell P, Knight PG, Boland MP, Roche JF. Effect of dominant follicle persistence
on follicular fluid oestradiol and inhibin and on oocyte maturation in heifers. J Reprod Fertil 1999;116:
293–304.
[31] Verkerk GA, Taufa VK, Morgan S, Clark BA, Macmillan KL. Effects of oestradiol benzoate by injection at
CIDR insertion for the treatment of postpartum anovulatory anoestrus in dairy cows. Proc NZ Soc Anim Prod
1998;58:82–4.
[32] Xu ZZ, Burton LJ, McDougall S, Jolly PD. Treatment of noncyclic lactating dairy cows with progesterone
and oestradiol or with progesterone, GnRH, prostaglandin F2a and estradiol. J Dairy Sci 2000;83:464–70.
[33] Rhodes FM, Burke CR, Clark BA, Day ML, Macmillan KL. Effect of treatment with progesterone and
oestradiol benzoate on ovarian follicular turnover in postpartum anoestrous cows and cows which have
resumed oestrous cycles. Anim Reprod Sci 2002;69:139–50.
[34] Shaham-Albalancy A, Rosenberg M, Folman Y, Graber Y, Meidan R, Wolfenson D. Two methods of
inducing low plasma progesterone concentrations have different effects on dominant follicles in cows. J
Dairy Sci 2000;83:2771–8.
[35] Echternkamp SE, Ferrell CL, Rone JD. Influence of pre- and post-partum nutrition on LH secretion in
suckled postpartum beef heifers. Theriogenology 1992;18:283–95.
[36] Wright IA, Rhind SM, Whyte TK, Smith AJ. Effects of body condition at calving and feeding level after
calving on LH profiles and the duration of the post-partum anoestrous period in beef cows. Anim Prod
1992;55:41–6.
[37] Ursely J, Leymarie P. Varying response to luteinizing hormone of two luteal cell types isolated from bovine
corpus luteum. J Endocrinol 1979;83:303–10.
[38] Gombe S, Hansel W. Plasma luteinizing hormone (LH) and progesterone levels in heifers on restricted
energy intakes. J Anim Sci 1973;37:728–33.
[39] Lucy MC, Collier RJ, Kitchell ML, Dibner JJ, Hauser SP, Kriui GG. Immunohistochemical and nucleic acid
analysis of somatotropin receptor populations in the bovine ovary. Biol Reprod 1993;48:1219–27.
[40] Juengel JL, Niswender GD. Molecular regulation of luteal progesterone synthesis in domestic ruminants. J
Reprod Fertil Suppl 1999;54:193–205.
[41] Liebermann J, Schams D. Actions of somatotrophin on oxytocin and progesterone release from
microdialysed bovine corpus luteum in vitro. J Endocrinol 1994;143:243–50.
[42] Lucy MC. Reproductive loss in high-producing dairy cattle: where will it end? J Dairy Sci 2001;84:
1277–93.
[43] Rhodes FM, McDougall S, Morgan SR, Verkerk GA. Supplementing treated anoestrous dairy cows with
progesterone does not increase conception rates. NZ Vet J 2001;49:8–12.
[44] Helmer SD, Britt JH. Hormone secretion and characteristics of estrous cycles after treatment of heifers with
human chorionic gonadotropin or prostaglandin F2 alpha during corpus luteum formation. J Anim Sci
1987;64:782–9.
[45] Schmitt EJ, Diaz T, Barros CM, de la Sota RL, Drost M, Fredriksson EW, et al. Differential response of the
luteal phase and fertility in cattle following ovulation of the first-wave follicle with human chorionic
gonadotropin or an agonist of gonadotropin-releasing hormone. J Anim Sci 1996;74:1074–83.
1548 S. McDougall et al. / Theriogenology 63 (2005) 1529–1548

[46] Santos JEP, Thatcher WW, Pool L, Overton MW. Effect of human chorionic gonadotropin on luteal
function and reproductive performance of high-producing lactating Holstein dairy cows. J Anim Sci 2001;
79:2881–94.
[47] Loeffler SH, de Vries MJ, Schukken YH. The effects of time of disease occurrence, milk yield, and body
condition on fertility of dairy cows. J Dairy Sci 1999;82:2589–604.
[48] McDougall S, Loeffler SH. Resynchrony of postpartum dairy cows previously treated for anestrus.
Theriogenology 2003;61:239–53.
[49] Heuer C, Schukken YH, Dobbelaar P. Postpartum body condition score and results from the first test day milk
as predictors of disease, fertility, yield, and culling in commercial dairy herds. J Dairy Sci 1999;82:295–304.
[50] Morton J. The InCalf project. Progress Report No. 2. Dairy Research and Development Corporation,
Melbourne, Australia, 2000.
[51] Buckley F, O’Sullivan K, Mee JF, Evans RD, Dillon P. Relationships among milk yield, body condition, cow
weight, and reproduction in spring-calved Holstein-Friesians. J Dairy Sci 2003;86:2308–19.