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Theriogenology 71 (2009) 1333–1342

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Review
Postpartum anestrus in dairy cattle
A.T. Peter a,*, P.L.A.M. Vos b, D.J. Ambrose c
a
Department of Veterinary Clinical Sciences, School of Veterinary Medicine, Purdue University, West Lafayette, IN 47907, USA
b
Department of Farm Animal Health, Faculty of Veterinary Medicine, Utrecht University, Utrecht, The Netherlands
c
Agriculture Research Division, Alberta Agriculture and Rural Development, Edmonton, Alberta T6H 5T6, Canada
Received 18 June 2008; accepted 14 November 2008

Abstract
Fertility of the postpartum period is negatively influenced by the incidence of anestrus. The latter condition is characterized by
the absence of estrous behavior, which may be an indication of suboptimal conditions (e.g., inadequate peripartum nutrition) or
pathologic conditions (e.g., chronic debilitating diseases or uterine and ovarian diseases). Although initiation of ovarian follicular
growth in the postpartum period is generally not affected, subsequent development (deviation) and the fate of the dominant follicle
are the primary factors that affect reestablishment of ovarian cyclicity. Anestrus can be classified based on the three functional states
of follicular development; that is, follicle emergence, deviation, and ovulation. Prevention of anestrus is preferable to treatment and
can be achieved in part by maintaining a healthy periparturient period. To better understand the etiology of anestrus and its
prevention, research is urgently needed in the following three areas: the role of peripartum disease conditions that influence
reproduction, genes involved in ovulation, and the influence of proteins (e.g., leptin) that appear to be important links between
metabolic signals and the neuroendocrine axis.
# 2009 Elsevier Inc. All rights reserved.

Keywords: Dairy cow; Postpartum; Anestrus; Types of anestrus

Contents

1. Introduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1334
2. Puerperium and resumption of ovarian activity . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1334
3. Energy balance and postpartum reproduction . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1335
4. Anestrus and its types . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1335
5. Managing anestrus . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1337
6. Prevention. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1339
7. Future directions . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1339
8. Conclusions. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1340
References . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 1340

* Corresponding author. Tel.: +1 765 494 5808 fax: +1 765 496 1108.
E-mail address: petera@purdue.edu (A.T. Peter).

0093-691X/$ – see front matter # 2009 Elsevier Inc. All rights reserved.
doi:10.1016/j.theriogenology.2008.11.012
1334 A.T. Peter et al. / Theriogenology 71 (2009) 1333–1342
1. Introduction
The postpartum period plays a pivotal role in cattle
reproduction. The duration of postpartum anestrus has
an important influence on reproductive performance
[1]. It has been suggested that in high-yielding dairy
herds, there is increased incidence of anestrus [2–6].
Perhaps increased partitioning of energy to milk
production can result in anestrus by delaying resump-
tion of follicular activity. However, factors such as
limited energy intake, lower body reserves, and
postpartum diseases can also delay the return to
cyclicity. A trouble-free calving predisposes to prompt
resumption of postpartum ovarian activity [6]. Ideally,
this should be followed by a minimal period of negative
energy balance (NEB). The duration of postpartum
anestrus is not determined by emergence of follicular
waves, but rather by follicular deviation and/or the fate
of the dominant follicle. Under optimal conditions,
there is deviation in follicular growth, selection of a
dominant follicle, its growth to maturity, ovulation, and
subsequently luteolysis, resulting in reestablishment of
cyclical ovarian activity, with an opportunity for
becoming pregnant. Failure of any of these processes
prolongs postpartum anestrus. In addition, failure of
cows to express estrus and poor estrus detection
efficiency can increase the incidence of anestrus in a
herd; however, this review will not focus on these two
conditions.
Historically, anestrus was broadly classified into
physiologic and pathologic (clinical) types, with the
following representing the pathologic type: (1) inactive
ovaries (i.e., minimal follicular development, anovula-
tion, and absence of a CL); (2) silent ovulation (i.e.,
ovulation without behavioral estrus); (3) ovarian
hypofunction (i.e., persistent dominant follicle); (4)
cystic ovarian degeneration (i.e., follicular or luteinized
follicular cyst); and (5) persistent CL (i.e., lack of luteal
regression) [7]. Recently, anestrus has been classified
based on ovarian follicular and luteal dynamics [8].
Follicular wave dynamics involve three main morpho-
logic events: emergence, deviation, and dominance,
ending in anovulation or ovulation. Therefore, classi-
fication of anestrus or anovulation based on follicle
characteristics at emergence, deviation, and dominance
provides for a rational diagnosis and treatment of the
underlying physiologic condition.
This review focuses on events leading to the
initiation and maintenance of ovarian activity as they
relate to postpartum anestrous conditions, including the
role of negative energy balance. It also attempts to build
on the recent classification of anestrus (based on
follicular and luteal dynamics [8]) and summarizes the
management (treatment) and prevention of true
anestrus. The proposed classification is not regarded
as definitive; rather, it is intended to provide additional
perspective in the ongoing discussion [8]. Future
advances in ovarian follicular dynamics and, in
particular, findings from microarray technology and
gene expression studies [9,10] will undoubtedly test the
validity of this classification.
2. Puerperium and resumption of ovarian
activity
During the puerperium, the uterus involutes and the
hypothalamo-hypophyseal-ovarian axis resumes cycli-
cal secretions of gonadotropic/gonadal hormones,
leading to first postpartum ovulations and regular
estrous cycles. In the normal puerperium, these events
are completed within 6 wk after calving. Ninety percent
of cows have their first postpartum ovulation within this
period [11]; however, the interval from calving to
ovulation can be 3 wk longer in the present-day dairy
cow [12].
Subclinical and clinical postpartum uterine infection
affect ovarian activity. Postpartum contamination of the
uterine lumen is inevitable; persistence of pathogenic
bacteria commonly causes clinical disease or subclini-
cal endometritis. It has been hypothesized that uterine
disease suppresses hypothalamic gonadotropin-releas-
ing hormone (GnRH) and possibly pituitary luteinizing
hormone (LH) secretion and has localized effects (i.e.,
delaying folliculogenesis) on ovarian function [13–17].
The mechanisms underlying the negative effects of
uterine infection on the reproductive tract may involve
the inflammatory response [18–20]. In cows with an
abnormal puerperium resulting in delayed uterine
involution, the resumption of ovarian activity is also
delayed. High circulating concentrations of prostaglan-
din F2a (PGF2a) in the first 3 wk postpartum due to
subclinical infection serve as a uterine signal, prevent-
ing premature onset of ovarian cyclicity until the
puerperal infection has been largely eliminated
[15,16,21]. In that regard, it is important for the uterus
to be free of infection to ensure functional/physiologic
follicular and luteal stages of the estrous cycle. Hence,
prolonged uterine secretion of PGF2a in cattle with
severe metritis has a role in prolonging postpartum
anestrus.
The anatomic and physiologic return of the genital
tract to the pregravid state and the events leading to the
optimal function of the hypothalamo-pituitary-ovarian
axis in a postpartum cow have an obligatory delay to
 A.T. Peter et al. / Theriogenology 71 (2009) 1333–1342
optimize fertility [22] and to direct energy toward milk
production [23]. Reestablishing pregnancy within 85 d
after calving requires supreme cooperation among the
involuting uterus, the hypothalamus, the pituitary, and
the ovaries, manifest as the resumption of ovarian
cyclicity, expression of estrus, and conception after a
timely breeding.
After parturition in dairy cows, sequential follicle-
stimulating hormone (FSH) increases (2 to 3 d duration)
are initiated in the first week postpartum [24], resulting
in emergence of the first postpartum follicular wave (3
to 5 follicles, 4 to 6 mm in diameter) within 10 to 14 d
after calving [25]. As FSH concentrations decline, one
follicle is selected for continued growth to become the
dominant follicle. This dominant follicle is believed to
suppress FSH secretion and the emergence of a new
wave, likely due to production of inhibin and estradiol.
The ability of this follicle to grow and ovulate depends
on the changes in the concentrations and the availability
of many growth factors within the follicle (e.g., insulin-
like growth factors and their binding proteins [26]), as
well as increased pulsatility of LH [27]. Another key
molecular event in the dominant follicle is differential
gene expression in granulosa cells, including increased
mRNA expression for LH receptor, 3-b hydroxysteroid
dehydrogenase, p450 side-chain cleavage, and p450
aromatase enzymes [28]. These changes within the
dominant follicle are necessary to stimulate steroido-
genesis, aromatase activity, and induce LH receptors on
granulosa cells. Ovulation, followed by the formation of
a CL, is a normal physiologic progression. In the
absence of ovulation, the dominant follicle either
becomes atretic or cystic; cows in which the dominant
follicle undergoes atresia have new follicular waves,
leading to the selection of new dominant follicles.
3. Energy balance and postpartum reproduction
Energy balance is the result of the energy ingested
(intake), minus the energy used (output) for main-
tenance and milk yield. Under normal prepartum
nutritional management, dairy cows rarely enter into
NEB prior to calving. After parturition, nutritional
requirements increase rapidly (due to milk production),
and most dairy cows enter a state of NEB. Cows in NEB
may preferentially divert nutrients away from repro-
duction [23,29,30], thereby limiting the number of
ovarian follicles, growth and maximum size of the
dominant follicle, delaying the first ovulation, interfer-
ing with ovulation, hindering the expression of estrus,
and lowering plasma progesterone concentrations. In
these cattle, ovulation is delayed by inhibition of LH
1335
pulse frequency and by suppressing blood concentra-
tions of glucose, insulin, and insulin-like growth factor-
1 (IGF-1), which reduce estrogen production by
dominant follicles [31].
In addition to energy deficits, increased feed intake
also suppresses reproduction by promoting steroid
hormone metabolism. Increased feed intake enhances
hepatic perfusion, thereby promoting clearance of
estradiol and progesterone [32], which can contribute
to anovulation [33], ovulation of larger dominant
follicles [34], multiple ovulations [35], poor luteal
function [36], and delayed luteal regression [6,12,37],
perhaps due to development of estrogen-inactive
dominant follicles [32,34], resulting in inadequate
endometrial PGF2a production. Furthermore, diets high
in crude protein, typically in excess of 16% to 17%,
support high milk yield but may be detrimental to
reproductive performance [38] due to elevated blood
urea concentrations [39].
4. Anestrus and its types
Anestrus is a broad term that indicates the lack of
expression of estrus (or absence of estrous signs),
despite efficient estrus detection. Anestrus is usually
characterized by a lack of ovarian progesterone
production. High-producing dairy cows have inherently
low expression of estrous signs [1], particularly during
the early postpartum period [40]. Therefore, overt signs
of estrus may not always precede an ovulation, in
particular the first postpartum ovulation [41]. Further-
more, in intensively managed modern dairy herds, the
efficiency of estrus detection can be extremely poor
[12], resulting in unobserved estrus. Thus, silent
ovulation (lack of overt signs of estrus) and unobserved
estrus (poor estrus detection efficiency [3]) can greatly
increase the incidence of anestrus. It is important to
distinguish between these conditions (physiology vs.
management). However, these two anestrous conditions
(silent ovulation and unobserved estrus) are not
discussed here because they are beyond the scope of
this review and have been described before [42,43].
Historically, anestrus was broadly classified into
physiologic and pathologic (clinical) types, with the
following four types representing the latter: silent
ovulation, cystic ovarian disease, ovarian hypofunction,
and ‘‘persistent’’ CL [7]. However, in this review, we
are attempting to classify anestrus in the context of
ovarian follicular and luteal dynamics.
The evaluation of ovarian follicular growth by
transrectal ultrasonography, coupled with measurement
of concentrations of reproductive hormones, has
1336 A.T. Peter et al. / Theriogenology 71 (2009) 1333–1342

Fig. 1. Schematic representation of types of anestrous conditions based on the physiology of ovarian follicular luteal dynamics. See the text for
details.

allowed designation of three functionally critical activity in the early postpartum period [41]. The
follicular diameters during follicular growth: emer- reduction in LH pulse frequency may be the result of
gence (4 mm), deviation (9 mm), and ovulation increases in the negative feedback effect of estradiol on
(variable, from 10 to 20 mm). Classification of LH pulse frequency. This can occur due to increased
anovulatory conditions on the basis of these three availability of estradiol receptors in the hypothalamus
critical points is logical, aids in rational diagnosis, and or increased sensitivity of hypothalamus to the negative
assists in treatment of the underlying physiologic feedback effect of estradiol, as well as other factors
condition [8]. The four types of anestrus discussed in [21]. In addition, there may be suppression of GnRH
this review are shown in Fig. 1. pulses, and it can be hypothesized that there is
In type I anestrus, there is growth of follicles to decreased GnRH neuronal activity (similar to pre-
emergence without further deviation or establishment of puberal anestrus).
a dominant follicle. The pathophysiology of this
condition is not well understood, but it is presumed
to be due to extreme undernutrition. In that regard,
undernutrition and severe energy deficit may cause this
condition through a lack of essential LH support to
sustain follicular growth and dominance [44]. Ovaries
associated with this type of anestrus may fall under the
classic description of ‘‘inactive ovaries’’ [45]. Two
ultrasonographic examinations of the ovaries, approxi-
mately 7 d apart, reveal no substantial changes in the
follicular structures, accompanied by a characteristic
absence of a CL or cystic follicular structures [46]. This
condition is expected to occur in <10% of the dairy
Fig. 2. Proposed current working model that summarizes the effects
population in the postpartum period in a normal herd of negative energy balance on reproductive parameters in the early
[46]. A working model (Fig. 2) was proposed to explain postpartum period (DF, dominant follicle; P4 = progesterone). Mod-
the effects of NEB on the resumption of follicular ified from Roche and Diskin [41].
 A.T. Peter et al. / Theriogenology 71 (2009) 1333–1342
In type II anestrus, there is deviation and growth,
followed by either atresia or regression. In certain cases,
the regression or atresia occurs only after a follicle has
reached a dominant status. Regression of this follicle
results in the emergence of a new follicular wave 2 to 3 d
later. In these cases, there are sequential follicular
waves prior to first ovulation, which may be delayed for
a prolonged interval. Some follicles grow further and
regress prior to ovulation [47]. These cows may have
low LH pulse frequency (<1 per 3 to 4 h). These
dominant follicles produce very low peripheral estradiol
concentrations; hence, there is either insufficient
estradiol production or failure of positive feedback
from basal estradiol production by the dominant
follicle. Subsequent follicular waves emerge within 1
to 2 d after this follicle regresses. It is noteworthy that
there may be up to nine waves of follicular growth
before first ovulation can occur [47].
In type III anestrus, there is deviation, growth, and
establishment of dominant follicle, but it fails to
ovulate and becomes a persistent follicular structure.
A single follicular structure >8 mm in diameter was
observed in the absence of a CL or cyst in two
ultrasonographic examinations 7 d apart [46,48]. This
may be due to insensitivity of the hypothalamus to the
positive feedback effect of estradiol or to altered
follicular responsiveness to gonadotrophic support,
mediated via metabolic hormones (e.g., insulin-like
growth factor and insulin) [24]. Persistent follicular
structures may become follicular cysts or they may
luteinize (luteal cysts). The latter occurs in 10% to
13% of the cases [49]. Follicular cysts may either
regress or persist as an anovulatory structure.
Depending on its structural/functional status, this
anovulatory structure may or may not suppress the
emergence of a subsequent postpartum follicular
wave for a variable interval [50].
Type IV anestrus is due to a prolonged luteal phase.
These cows have normal estrus, ovulation, and
formation of a CL, with prolonged luteal function
due to a lack of luteal regression. A contributing factor
may be the lack of an estrogenic dominant follicle at
the expected time of luteal regression [8]. In that
regard, estradiol from a dominant follicle is believed
to induce the formation of uterine oxytocin receptors,
leading to pulsatile release of PGF2a [51–53]. Many
factors have been suggested to increase the risk of a
prolonged luteal phase [6], including parity, dystocia,
health problems during the first month of lactation,
heat stress, and perhaps ovulation soon after calving.
Uterine infection [17] or pyometra can prolong the life
of the CL [54].
1337
5. Managing anestrus
For type I and type II anestrus conditions, remedial
measures involve correcting the NEB, which is very
difficult to achieve through nutritional management.
However, recent studies indicate that a reduction in the
length of dry period or its elimination [55] can attenuate
the detrimental effects of NEB and potentially reduce
anestrus. Furthermore, feeding certain dietary fatty
acids during the dry period (without altering its length)
can also reduce the interval from calving to the first
postpartum ovulation [56].
Much has been discussed regarding type I and type II
anovulatory noncystic anestrous conditions [6,57–61].
The prevalence ranged from 11% to 38% by 50 to 60 d
after calving [6,57]. Transrectal examination as a
diagnostic method has its limitations and can result in
misdiagnosis, leading to inappropriate administration of
hormones. Although transrectal ultrasonography has
been used to assess ovaries [60], the physiologic and
endocrine status of the ovarian structures is best
determined by measurement of progesterone concen-
trations [58].
If energy requirements meet the demands of produc-
tion, hormonal treatment can be used for these ‘‘static
follicles’’ [57]. In general, exogenous progestins are
considered appropriate for noncyclic or anestrous
postpartum cows [62]. The development of controlled
intravaginal progesterone-releasing devices, or an intra-
vaginal progesterone insert (IPI), hastened resumption of
ovarian follicular activity. The IPI facilitated hormone
treatments and circumvented delivery problems asso-
ciated with feeding or injecting progestins. At the end of
IPI treatment, hormones such as equine chorionic
gonadotropin (Fig. 3 [63]), estradiol, or PGF2a have
been given to maximize the response (resumption of
follicular activity). In a recent study [64], 64.2% (571 of
889) of cows classified as ‘‘subestrus or ovarian
hypofunction’’ group (akin to type I and type II anestrus
discussed in this review) responded to 9 d of IPI treatment
followed by an injection of PGF2a. Furthermore,
estradiol or GnRH analogues have been used prior to
IPI treatment [57]. Various combinations of GnRH
analogues and PGF2a have been used to initiate ovarian
cyclicity. Another approach is the use of timed-
insemination protocols (e.g., the Ovsynch program
[60]). In the various hormonal treatments listed above,
responses to treatments are not uniform either across
herds or across cows within herds; they appeared to be
dependent on factors influencing the prevalence of
anestrus (e.g., age, body condition, and postpartum
interval). Therefore, we concluded that no particular
1338 A.T. Peter et al. / Theriogenology 71 (2009) 1333–1342
Fig. 3. Use of IPI and equine chorionic gonadotropin (eCG) enables the cow to have required-size dominant follicle with adequate estradiol. This
allows the cow to exhibit estrous behavior, LH surge, followed by ovulation of a healthy oocyte (0 = time of deviation). Modified from Schmitt [63].
treatment can be recommended unequivocally for all
herds.
It is noteworthy that cows in the very early
postpartum period or those with low reserves of body
fat are unlikely to respond to hormonal treatments.
Although exogenous progestins are considered the most
appropriate therapy for noncyclic or anestrous (types I
and II) postpartum dairy cows, any hormonal method to
induce ovulation should be done in conjunction with
correction of management problems. Pretreatment of
anestrous cows with progesterone for 5 to 9 d is a
prerequisite for the concomitant expression of estrus at
first ovulation. However, the need for additional
exogenous hormones to ensure ovulation of a dominant
follicle is dependent on energy status, body condition
score, and the postpartum interval (which regulate LH
pulse frequency). The combination of an IPI and
estradiol have been reported [65]. In certain cases,
anestrous cows that failed to respond to the initial
treatment responded favorably to a second IPI treat-
ment. These cows received an IPI treatment for 6 d,
received 0.5 mg estradiol benzoate 1 d after the removal
of IPI, and were treated with another IPI for 6 d,
followed by its removal and treatment with 0.5 mg
estradiol benzoate on Day 7 [66].
Cows with persistent ovarian follicles can be
successfully treated with progesterone, GnRH, and
PGF2a and subsequently timed-inseminated [48].
Estradiol benzoate, given in association with proges-
terone, reduced the persistence of dominant follicles in
both cycling and anestrous cows but delayed subsequent
follicular development in some anestrous cows [67].
For follicular cysts, agents that induce the release of
LH from the anterior pituitary (e.g., GnRH) or have LH-
like action (e.g., hCG) can be used. Regardless of the
type of agents used, if cysts become luteinized, luteal
regression must be induced with PGF2a. A combination
of GnRH and PGF2a can also be used (Day 0, GnRH;
Day 7, PGF2a; and Day 9, GnRH), with or without
timed-insemination [68–70]. Other researchers have
treated cows with progesterone per se [71] or with
concurrent GnRH followed by PGF2a [69]. Progester-
one treatment can reestablish hypothalamic function
and normal cyclicity in cows with follicular cysts;
presumably, it can induce estrogen receptor a in the
mediobasal hypothalamus, thereby facilitating a GnRH/
LH surge in response to follicular estrogen [72]. The
incorporation of progesterone into a timed-insemination
protocol for reproductive management of cows with
cystic ovarian follicles has considerable potential [73]
and must be evaluated under field conditions to
determine its efficiency and usefulness. Before these
treatments can be recommended, further studies are
needed with larger herds to determine the economics of
their use, which has to be considered in relation to the
reduction in open interval in these cows [74]. It is
noteworthy that regulatory policies may prohibit the use
of these hormones in lactating dairy cows. For luteal
cysts, PGF2a or its analogues are the treatment of
choice; they induce regression of luteinized cysts, with
estrus occurring within 8 d in 87% to 96% of treated
cows [49].
For the fourth type of anestrus, PGF2a is used to
terminate the life of the CL and initiate the estrous
cycle. If the persistence of CL is due to pyometra,
treatment with PGF2a will also be beneficial [75]. In
certain cases, depending on the severity and amount of
collection of purulent material in the uterus, uterine
 A.T. Peter et al. / Theriogenology 71 (2009) 1333–1342
lavage and/or intrauterine or systemic antibiotic
treatments are recommended [3,76,77].
Although many hormonal treatments are possible for
certain anestrous conditions, the application of hormo-
nal therapy for the treatment of anestrus should be
approached with caution. It is inappropriate to use
hormones in an attempt to correct management
deficiencies. Furthermore, incorrect hormonal treat-
ment can transform a cow from one form of anestrus to
another [64]. To establish/modify hormone therapies,
factors affecting both the incidence of the different
types of anestrus and the estrus response after a specific
treatment should be continually monitored at the farm
level [64].
6. Prevention
High milk production and excellent fertility con-
tribute to the profitability of dairy herds. Although
reproductive performance of a high-producing dairy
cow has been questioned [78], there is evidence that
fertility is not necessarily impaired in cows with high
milk production [35,79], provided that the herd
environment is conducive and the peripartum health
of the animal is sustained [79]. Acceptable levels of
fertility can be achieved with cows of high genetic
merit, provided that estrus is detected efficiently, the
cows calve in moderate body condition, and they are fed
to minimize the loss of body condition after calving
[80].
Optimal reproductive function in the postpartum
period requires prevention of metabolic diseases in the
periparturient period [29] and rapid intake of substantial
dry matter after calving to meet the huge nutrient
demands of milk production. This process of metabolic
adaptation in early lactating cows should not be
hampered, as this can lead to suboptimal reproductive
function [81].
The first action must be to prevent deep NEB and the
development of hepatic lipidosis. Shortening or
eliminating the dry period may improve energy status
of cows and increase reproductive efficiency [55].
Overfatness at calving reduces appetite, thereby
affecting energy intake during the early postpartum
period. The consequent lipolysis increases plasma
nonesterified fatty acid (NEFA) concentration; uptake
of NEFA by the liver is proportional to NEFA
concentrations in the blood. Hepatic NEFA can either
be oxidized or esterified. The primary esterification
product is triglyceride, which can either be exported as
part of a very low density lipoprotein or stored. In
ruminants, export occurs at a very slow rate relative to
1339
many other species. Therefore, under conditions of
elevated hepatic NEFA uptake and esterification,
triglycerol (TAG) accumulates. Because of inadequate
lipoprotein synthesis, TAGs are accumulated in liver
[82,83]. Hepatic lipidosis may not have a direct effect
on the function of the hypothalamo-utero-ovarian axis,
but it can reduce the immune competence [84],
increasing the incidence of infections and metabolic
diseases. For example, a higher incidence of metritis
was observed when fat mobilization was pronounced
[85]. Regarding the controversy between milk yield and
fertility, it is noteworthy that increased milk production
has no negative effect on fertility, as long as the amount
of TAG in the liver remains consistent [86]. Although
the detrimental effect of elevated levels of NEFA is not
known, a recent study suggested that postpartum cows
with ovulatory dominant follicles had lower plasma
NEFA compared with that of nonovulatory dominant
follicles [87]. Further research is needed to determine
the exact role of TAG and NEFA in the function of the
hypothalamo-utero-ovarian axis. In summary, it is
important to pay attention to heat stress [88], manage-
ment factors [31], high producers [4], abnormal
puerperium [89], metabolic diseases [29], pasture
intake [47], nutritional restrictions [90], and finally
uterine conditions (e.g., endometritis) [17].
7. Future directions
The LH-induced transition of a preovulatory follicle
to one that can ovulate is a complex multigene,
multistep process in which timing is of the essence.
Some of the genes involved in the process are known in
laboratory animals [91], and similar work has to be
conducted in cattle. Recent work identified novel genes
associated with dominant follicle development in cattle
[9,10]. Many genes involved in the process of follicular
development, selection, and ovulation remain to be
documented to elucidate the control of ovulation and the
molecular pathology associated with anovulation and
anestrus.
Another area of future study is the influence of leptin
in the resumption of ovarian follicular activity in the
postpartum period, in particular its potential role in feed
intake [92,93] and perhaps in the formation of follicular
cysts [94]. Substantially more studies are needed to
elucidate interactions between nutrition and reproduc-
tion in dairy cows, particularly between the somato-
tropic and gonadotropic axes [95].
Progress toward understanding the mechanisms
behind the effects of disease on reproduction has been
slow in the past; to understand this relationship
1340 A.T. Peter et al. / Theriogenology 71 (2009) 1333–1342
completely, research in this important area should
involve veterinarians and animal scientists [96].
8. Conclusions
During the past 50 years, there has been considerable
advancement in our ability to intervene in the
reproduction of cattle. However, fertility in dairy cattle
has consistently declined, despite increasing veterinary
intervention [54]. A major factor that determines the
initiation of ovarian activity and allows the animal to
maintain cyclicity is postpartum energy balance. There
may be other causes (e.g., postpartum uterine diseases
leading to anestrus). It is more effective to prevent
rather than treat anestrus. Although the appropriate use
of hormones does not have any known negative effects
on animal welfare or public health, the concern of
consumers regarding treatments must be considered
[97]. We need to focus on improving animal health and
fertility by promoting good management rather than by
relying on the widespread use of exogenous hormones.
High reproductive efficiency in the postpartum period
requires the prevention of metabolic diseases in the
periparturient period and high dry-matter intake after
calving to meet the demands of milk production.
Further research is needed in the area of postpartum
reproduction, particularly with respect to the molecular
pathology associated with anovulation that leads to
anestrus.
References
[1] Lucy MC. Fertility in high-producing dairy cows: reasons for
decline and corrective strategies for sustainable improvement.
Soc Reprod Fertil Suppl 2007;64:237–54.
[2] Berger BJ, Shanks RD, Freeman AE, Laben RC. Genetic aspects
of milk yield and reproductive performance. J Dairy Sci
1981;64:114–22.
[3] Etherington WG, Christie KA, Walton JS, Leslie KE, Wickstrom
S, Johnson WH. Progesterone profiles in postpartum Holstein
dairy cows as an aid in the study of retained fetal membranes,
pyometra and anestrus. Theriogenology 1991;35:731–46.
[4] Opsomer G, Mijten P, Coryn M, de Kruif A. Postpartum anoes-
trus in dairy cows: a review. Vet Q 1996;18:68–75.
[5] Opsomer G, Coryn M, Deluyker H, de Kruif A. An analysis of
ovarian dysfunction in high-yielding dairy cows after calving
based on progesterone profiles. Reprod Domest Anim 1998;33:
193–204.
[6] Opsomer G, Grohn YT, Hertl J, Coryn M, Deluyker H, de Kruif
A. Risk factors for post partum ovarian dysfunction in high
producing dairy cows in Belgium: a field study. Theriogenology
2000;53:841–57.
[7] Mwaanga ES, Janowski T. Anoestrus in dairy cows: causes,
prevalence and clinical forms. Reprod Domest Anim 2000;35:
193–200.
[8] Wiltbank MC, Gumen A, Sartori R. Physiological classification
of anovulatory conditions in cattle. Theriogenology 2002;57:
21–52.
[9] Zielak AE, Forde N, Park SD, Doohan F, Coussens PM, Smith GW.
Identification of novel genes associated with dominant follicle
development in cattle. Reprod Fertil Dev 2007;19:967–75.
[10] Evans AC, Forde N, O’Gorman GM, Zielak AE, Lonergan P, Fair
T. Use of microarray technology to profile gene expression
patterns important for reproduction in cattle. Reprod Domest
Anim 2008;43:359–67.
[11] Peter AT, Bosu WTK. Postpartum ovarian activity in dairy cows:
correlation between behavioral estrus; pedometer measurements
and ovulations. Theriogenology 1986;26:111–5.
[12] Ambrose DJ, Colazo MG. Reproductive status of dairy herds in
Alberta: a closer look. Proceedings of the 2007 Western Cana-
dian Dairy Seminar. Adv Dairy Technol 2007;19:227–44.
[13] Bosu WT, Peter AT. Evidence for a role of intrauterine infections
in the pathogenesis of cystic ovaries in postpartum dairy cows
1987;28:725–36.
[14] Bosu WT, Peter AT, DeDecker RJ. Short-term changes in serum
luteinizing hormone, ovarian response and reproductive perfor-
mance following gonadotrophin releasing hormone treatment in
postpartum dairy cows with retained placenta. Can J Vet Res
1988;52:165–71.
[15] Peter AT, Bosu WTK. Relationship of uterine infections and
folliculogenesis in dairy cows during early puerperium. Ther-
iogenology 1988;30:1045–51.
[16] Peter AT, Bosu WTK, Gilbert RO. Absorption of Escherichia
coli endotoxin (lipopolysaccharide) from the uteri of postpartum
dairy cows. Theriogenology 1990;33:189–94. 1011–1014; 34.
[17] Mateus L, da Costa LL, Bernardo F, Silva JR. Influence of
puerperal uterine infection on uterine involution and postpartum
ovarian activity in dairy cows. Reprod Domest Anim 2002;37:
31–5.
[18] Sheldon IM, Dobson H. Postpartum uterine health in cattle.
Anim Reprod Sci 2004;82-83:295–306.
[19] Herath S, Dobson H, Bryant CE, Sheldon IM. Use of the cow as a
large animal model of uterine infection and immunity. J Reprod
Immunol 2006;69:13–22.
[20] Williams EJ, Fischer DP, Noakes DE, England GC, Rycroft A,
Dobson H, et al. The relationship between uterine pathogen
growth density and ovarian function in the postpartum dairy cow.
Theriogenology 2007;68:549–59.
[21] Sheldon IM, Noakes DE, Rycroft AN, Pfeiffer DU, Dobson H.
Influence of uterine bacterial contamination after parturition on
ovarian dominant follicle selection and follicle growth and
function in cattle. Reproduction 2002;123:837–45.
[22] Bilodeau-Goeseels S, Kastelic JP. Factors affecting embryo
survival and strategies to reduce embryonic mortality in cattle.
Can J Anim Sci 2003;83:659–71.
[23] Bauman DE, Currie WB. Partitioning of nutrients during preg-
nancy and lactation: a review of mechanisms involving home-
ostasis and homeorhesis. J Dairy Sci 1980;63:1514–29.
[24] Beam SW, Butler WR. Effects of energy balance on follicular
development and first ovulation in postpartum dairy cows. J
Reprod Fertil Suppl 1999;54:411–24.
[25] Rajamahendran R, Taylor C. Characterization of ovarian activity
in postpartum dairy cows using ultrasound imaging and proges-
terone profiles. Anim Reprod Sci 1990;22:171–80.
[26] Fortune JE, Rivera GM, Yang MY. Follicular development: the
role of the follicular microenvironment in selection of the
dominant follicle. Anim Reprod Sci 2004;82–83:109–26.
 A.T. Peter et al. / Theriogenology 71 (2009) 1333–1342
[27] Canfield RW, Butler WR. Energy balance and pulsatile LH
secretion in early postpartum dairy cattle. Domest Anim Endo-
crinol 1990;7:323–30.
[28] Bao B, Garverick HA. Expression of steroidogenic enzyme and
gonadotropin receptor genes in bovine follicles during ovarian
follicular waves: a review. J Anim Sci 1998;76:1903–21.
[29] Ferry J. Clinical Management of Anestrus. In: Youngquist RS,
editor. Current Therapy in Large Animal Theriogenology. WB
Saunders; 1997. p. 285–9.
[30] Leroy JL, Vanholder T, Van Knegsel AT, Garcia-Ispierto I, Bols
PE. Nutrient prioritization in dairy cows early postpartum:
mismatch between metabolism and fertility? Reprod Domest
Anim 2008;43:96–103.
[31] Butler WR. Nutritional interactions with reproductive perfor-
mance in dairy cattle. Anim Reprod Sci 2000;60–61:449–57.
[32] Sangsritavong S, Combs DK, Sartori R, Armentano LE, Wilt-
bank MC. High feed intake increases liver blood flow and
metabolism of progesterone and estradiol-17b in dairy cattle.
J Dairy Sci 2002;85:2831–42.
[33] Walsh RB, Kelton DF, Duffield TF, Leslie KE, Walton JS,
LeBlanc SJ. Prevalence and risk factors for postpartum anovu-
latory condition in dairy cows. J Dairy Sci 200X;90:315-324.
[34] Sartori R, Haughian JM, Shaver RD, Rosa GJM, Wiltbank MC.
Comparison of ovarian function and circulating steroids in
estrous cycles of Holstein heifers and lactating cows. J Dairy
Sci 2004;87:905–20.
[35] Lopez H, Caraviello DZ, Satter LD, Fricke PM, Wiltbank MC.
Relationship between level of milk production and multiple
ovulations in lactating dairy cows. J Dairy Sci 2005;88:2783–93.
[36] Villa-Godoy A, Hughes TL, Emery RS, Chapin LT, Fogwell RL.
Association between energy balance and luteal function in
lactating dairy cows. J Dairy Sci 1988;71:1063–72.
[37] Petersson K-J, Gustafsson H, Strandberg E, Berglund B. Aty-
pical progesterone profiles and fertility in Swedish dairy cows. J
Dairy Sci 2006;89:2529–38.
[38] Tamminga S. The effect of the supply of rumen degradable
protein and metabolisable protein on negative energy balance
and fertility in dairy cows. Anim Reprod Sci 2006;96:227–39.
[39] Butler WR. Nutritional effects on resumption of ovarian cycli-
city and conception rate in postpartum dairy cows. Occ Publ Br
Soc Anim Sci 2001;26:133–45.
[40] Harrison RO, Ford SP, Young JW, Conley AJ, Freeman AE.
Increased milk production versus reproductive and energy status
of high producing dairy cows. J Dairy Sci 1990;73:2749–58.
[41] Roche JF, Diskin MD. Resumption of reproductive activity in the
early postpartum period of cows. Occ Publ Br Soc Anim Sci
2001;26:31–42.
[42] Allrich RD. Estrous behavior and estrus detection in cattle. Vet
Clin North Am Food Anim Pract 1993;9:249–62.
[43] Allrich RD. Endocrine and neural control of estrus in dairy cows.
J Dairy Sci 1994;77:2738–44.
[44] Jolly PD, McDougall S, Fitzpatrick LA, Macmillan KL, Entwis-
tle KW. Physiological effects of undernutriotn on postpartum
anoestrous cows. J Reprod Fertil 1995;49:477–92.
[45] Fielden ED, Harris RE, Macmillan KL, Shrestha SL. Some
aspects of reproductive performance in selected town-supply
dairy herds. NZ Vet J 1980;131–132:141–2.
[46] Markusfeld O. Inactive ovaries in high-yielding dairy cows
before service: aetiology and effect on conception. Vet Rec
1987;121:149–53.
[47] McDougall S, Burke CR, MacMillan KL, Williamson NB.
Patterns of follicular development during periods of anovulation
1341
in pasture-fed dairy cows after calving. Res Vet Sci 1995;58:
212–6.
[48] Lopez-Gatius F, Santolaria P, Yaniz J, Rutllant J, Lopez-Bejar M.
Persistent ovarian follicles in dairy cows: a therapeutic approach.
Theriogenology 2001;56:649–59.
[49] Peter AT. An update on cystic ovarian degeneration in cattle.
Reprod Domest Anim 2004;39:1–7.
[50] Sakaguchi M, Sasamoto Y, Suzuki T, Takahashi Y, Yamada Y.
Fate of cystic ovarian follicles and the subsequent fertility of
early postpartum dairy cows. Vet Rec 2006;159:197–201.
[51] Thatcher WW, Macmillan KL, Hansen PJ, Drost M. Concepts
for regulation of corpus luteum function by the conceptus and
ovarian follicles to improve fertility. Theriogenology 1989;31:
149–64.
[52] McCracken JA, Custer EE, Lamsa JC. Luteolysis: a neuroendo-
crine-mediated event. Physiol Rev 1999;79:263–323.
[53] Knickerbocker JJ, Thatcher WW, Foster DB, Wolfenson D,
Bartol FF, Caton D. Uterine prostaglandin and blood flow
responses to estradiol-17b in cyclic cattle. Prostaglandins
1986;31:757–76.
[54] Sheldon IM, Wathes DC, Dobson H. The management of bovine
reproduction in elite herds. Vet J 2006;171:70–8.
[55] Grummer RR. Strategies to improve fertility of high yielding
dairy farms: management of the dry period. Theriogenology
2007;68:5281–8.
[56] Colazo MG, Ambrose DJ, Hayirli A, Doepel L. Prepartum feed
restriction and fatty acid supplementation influence reproductive
performance of dairy cows. J Dairy Sci 2007;90:328.
[57] Rhodes FM, McDougall S, Burke CR, Verkerk GA, Macmillan
KL. Invited review: treatment of cows with an extended post-
partum anestrous interval. J Dairy Sci 2003;86:1876–94.
[58] Silva E, Sterry RA, Fricke PM. Assessment of a practical
method for identifying anovular dairy cows synchronized for
first postpartum timed artificial insemination. J Dairy Sci 2007;
90:3255–62.
[59] Walsh RB, Leblanc SJ, Duffield TF, Kelton DF, Walton JS,
Leslie KE. The effect of a progesterone releasing intravaginal
device (PRID) on pregnancy risk to fixed-time insemination
following diagnosis of non-pregnancy in dairy cows. Therio-
genology 2007;67:948–56.
[60] Stevenson JS, Tenhouse DE, Krisher RL, Lamb GC, Larson JE,
Dahlen CR, et al. Detection of anovulation by heatmount
detectors and transrectal ultrasonography before treatment with
progesterone in a timed insemination protocol. J Dairy Sci
2008;9:2901–15.
[61] Petersson KJ, Strandberg E, Gustafsson H, Royal MD, Berglund
B. Detection of delayed cyclicity in dairy cows based on
progesterone content in monthly milk samples. Prevent Vet
Med 2008;86:153–63.
[62] Yániz JL, Murugavel K, López-Gatius F. Recent developments
in oestrous synchronization of postpartum dairy cows with and
without ovarian disorders. Reprod Domest Anim 2004;39:86–93.
[63] Schmitt EJ. Overview of dairy reproductive physiology. In:
Presented at: European Dairy Symposium; 2000.
[64] López-Gatius F, Mirzaei A, Santolaria P, Bech-Sàbat G, Nogar-
eda C, Garcia-lspierto I, et al. Factors affecting the response to
the specific treatment of several forms of clinical anestrus in high
producing dairy cows. Theriogenology 2008;69:1095–103.
[65] McDougall S, Compton CW, Anniss FM. Effect of exogenous
progesterone and oestradiol on plasma progesterone concentra-
tions and follicle wave dynamics in anovulatory anoestrous post-
partum dairy cattle. Animal Reprod Sci 2004;84:303–14.
1342 A.T. Peter et al. / Theriogenology 71 (2009) 1333–1342
[66] McDougall S, Loeffler SH. Resynchrony of postpartum dairy
cows previously treated for anestrus. Theriogenology 2004;61:
239–53.
[67] 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.
[68] Bartolome JA, Archbald LF, Morresey P, Hernandez J, Tran T,
Kelbert D, et al. Comparison of synchronization of ovulation and
induction of estrus as therapeutic strategies for bovine ovarian
cysts in the dairy cow. Theriogenology 2000;53:815–25.
[69] Ambrose DJ, Schmitt EJ-P, Lopes FL, Mattos RC, Thatcher
WW. Ovarian and endocrine responses associated with the
treatment of cystic ovarian follicles in dairy cows with gonado-
tropin releasing hormone and prostaglandin F2a, with or without
exogenous progesterone. Can Vet J 2004;45:931–7.
[70] Crane MB, Melendez P, Bartolome J, de Vries A, Risco C,
Archbald LF. Association between milk production and treat-
ment response of ovarian cysts in lactating dairy cows using the
Ovsynch protocol. Theriogenology 2006;66:1243–8.
[71] Hatler TB, Hayes SH, Anderson LH, Silvia WJ. Effect of single
injection of progesterone on ovarian follicular cysts in lactating
dairy cows. Vet J 2006;172:329–33.
[72] Gümen A, Wiltbank MC. An alteration in the hypothalamic
action of estradiol due to lack of progesterone exposure can
cause follicular cysts in cattle. Biol Reprod 2002;66:1689–95.
[73] Crane MB, Bartolome J, Melendez P, de Vries A, Risco C,
Archbald LF. Comparison of synchronization of ovulation with
timed insemination and exogenous progesterone as therapeutic
strategies for ovarian cysts in lactating dairy cows. Theriogen-
ology 2006;65:1563–74.
[74] De Vries A, Crane MB, Bartolome JA, Melendez P, Risco CA,
Archbald LF. Economics of comparison of timed artificial
insemination and exogenous progesterone as treatments for
ovarian cysts. J Dairy Sci 2006;89:3028–37.
[75] Farin PW, Estill CT. Infertility due to abnormities of the ovaries
in cattle. Vet Clin North Am Food Anim Pract 1993;9:291–308.
[76] Fazeli M, Ball L, Olson JD. Comparison of treatment of pyo-
metra with estradiol cypionate or cloprostenol followed by
infusion or non-infusion with nitrofurazone. Theriogenology
1980;14:339–47.
[77] Földi J, Kulcsár M, Pécsi A, Huyghe B, de Sa C, Lohuis JA, Cox
P, et al. Bacterial complications of postpartum uterine involution
in cattle. Anim Reprod Sci 2006;96:265–81.
[78] Dobson H, Smith R, Royal M, Knight CH, Sheldon I. The high-
producing dairy cow and its reproductive performance. Reprod
Domest Anim 2007;42:17–23.
[79] Windig JJ, Calus MP, Veerkamp RF. Influence of herd environ-
ment on health and fertility and their relationship with milk
production. J Dairy Sci 2005;88:335–47.
[80] Mayne CS, McCoy MA, Lennox SD, Mackey DR, Verner M,
Catney DC, et al. Fertility of dairy cows in Northern Ireland. Vet
Rec 2002;150:707–13.
[81] Jorritsma R, Wensing T, Kruip TA, Vos PL, Noordhuizen JP.
Metabolic changes in early lactation and impaired reproductive
performance in dairy cows. Vet Res 2003;34:11–26.
[82] Roberts CJ, Reid IM, Rowlands GJ, Patterson A. A fat mobilisa-
tion syndrome in dairy cows in early lactation. Vet Rec 1981;
108:7–9.
[83] van den Top AM, Wensing T, Geelen MJ, Wentink GH, van’t
Klooster AT, Beynen AC. Time trends of plasma lipids and
enzymes synthesizing hepatic triacylglycerol during postpartum
development of fatty liver in dairy cows. J Dairy Sci 1995;
78:2208–20.
[84] Wentink GH, Rutten VPMG, van den Ingh TSGAM, Muller
KE, Wensing T. Impaired specific immunoreactivity in cows
with hepatic lipidosis. Vet Immunol Immunopathol 1997;56:
77–83.
[85] Kaneene JB, Miller R, Herdt TH, Gardiner JC. The association of
serum nonesterified fatty acids and cholesterol, management and
feeding practices with peripartum disease in dairy cows. Prev Vet
Med 1997;31:59–72.
[86] Jorritsma R, Jorritsma H, Schukken YH, Wentink GH. Relation-
ships between fatty liver and fertility and some periparturient
diseases in commercial Dutch dairy herds. Theriogenology
2000;54:1065–74.
[87] Butler ST, Pelton SH, Knight PG, Butler WR. Follicle-stimulat-
ing hormone isoforms and plasma concentrations of estradiol
and inhibin A in dairy cows with ovulatory and non-ovulatory
follicles during the first postpartum follicle wave. Domest Anim
Endocrinol 2008;35:112–9.
[88] Roth Z, Meidan R, Shaham-Albalancy A, Braw-Tal R, Wolfen-
son D. Delayed effect of heat stress on steroid production in
medium-sized and preovulatory bovine follicles. Reproduction
2001;121:745–51.
[89] Kruip TAM, Wensing T, Vos PLAM. Characteristics of abnormal
puerperium in dairy cattle and the rationale for common treat-
ments. Occ Publ Br Soc Anim Sci 2001;26:63–79.
[90] Roberts AJ, Nugent 3rd RA, Klindt J, Jenkins TG. Circulating
insulin-like growth factor I, insulin-like growth factor binding
proteins, growth hormone, and resumption of estrus in postpar-
tum cows subjected to dietary energy restriction. J Anim Sci
1997;75:1909–17.
[91] Robker RL, Russell DL, Yoshioka S, Sharma SC, Lydon JP,
O’Malley BW, et al. Ovulation: a multi-gene, multi-step process.
Steroids 2000;65:559–70.
[92] Spicer LJ. Leptin: a possible metabolic signal affecting repro-
duction. Domest Anim Endocrinol 2001;21:251–70.
[93] Reist M, Erdin D, von Euw D, Tschuemperlin K, Leuenberger H,
Delavaud C, et al. Concentrate feeding strategy in lactating dairy
cows: metabolic and endocrine changes with emphasis on leptin.
J Dairy Sci 2003;86:1690–706.
[94] Smith GD, Jackson LM, Foster DL. Leptin regulation of
reproductive function and fertility. Theriogenology 2002;57:
73–86.
[95] Chagas LM, Bass JJ, Blache D, Burke CR, Kay JK, Lindsay DR,
et al. Invited review: new perspectives on the roles of nutrition
and metabolic priorities in the subfertility of high-producing
dairy cows. J Dairy Sci 2007;90:4022–32.
[96] Lucy MC. Reproductive loss in high-producing dairy cattle:
where will it end? J Dairy Sci 2001;84:1277–93.
[97] Refsal AO. To treat or not to treat: a proper use of hormones and
antibiotics. Anim Reprod Sci 2000;60–61:109–19.