Reprod Dom Anim 47 (Suppl. 3), 34–44 (2012); doi: 10.1111/j.1439-0531.2012.02032.
x
ISSN 0936-6768
Use of Ultrasound in the Reproductive Management of Dairy Cattle
LA Quintela, M Barrio, AI Peña, JJ Becerra, J Cainzos, PG Herradón and C Dı́az
Faculty of Veterinary Medicine, University of Santiago de Compostela, Lugo, Spain
Contents
The significant decrease in fertility observed in dairy cattle
during the last few decades and increasing requirements by the
farmers have made a regular control of reproduction indis-
pensable to urgently identify and solve potential problems
affecting reproductive efficiency. Traditionally, the main diag-
nostic methods used for reproductive control in cattle included
rectal palpation, inspection of vaginal discharge and vaginos-
copy. Since the 1990s, the use of ultrasound (US) has become a
common diagnostic method as a result of the new advances
made in the development of US scans: smaller size, high level
of autonomy, high image quality and accessible prices.
Ultrasound improves accuracy in the diagnoses of stages of
the oestrous cycle, ovarian and uterine pathologies, and
pregnancy diagnosis. In addition, it facilitates the diagnosis
of alterations during pregnancy (embryo mortality, foetal
malformations, etc.) and helps determining foetal sex from day
55 of pregnancy.
Introduction
The use of ultrasound (US) as a diagnostic tool for
reproductive control in cattle was not very common
until the mid-1990s, when small portable, affordable
scanners, with a good quality image first became
available on the market. Although US has been used
in veterinary practice for more than 30 years (Horder
et al. 1996), especially on small animals and horses, its
application on cattle was limited because of the difficulty
of moving the heavy low-autonomy US scanners around
cattle farms.
During the last ten to fifteen years, the advent of
lighter and smaller US machines on the market has
encouraged the use of US by cattle practitioners more
and more, and consequently, the increasing demand for
equipment and knowledge has in turn stimulated the
development of ever-improving US equipment. As a
result, portable US scanners currently available on the
market are of excellent quality, provide a high degree of
autonomy and are reasonably priced, being fully suit-
able for cattle practitioners.
As field practitioners were acquiring more knowledge
and expertise in the use of US, they progressively
abandoned the erroneous concept that the main advan-
tage, if not the only one, of using US on cows was the
possibility of early pregnancy diagnosis. The main appli-
cations of ultrasonography in the field of cattle reproduc-
tion will be reviewed in this paper and, when appropriate,
its accuracy compared with that of rectal palpation.
Exploration of the Uterus in the Non-Pregnant
Cow
The exploration of the uterus in the non-pregnant
female aims to either determine the stage of the oestrous
cycle of the cow or diagnose pathologic conditions of
the uterus (Beal et al. 1992). The histology of the uterus
determines its ecographic appearance, as the submucosa
can be visualized as an anechoic layer beneath the
endometrium. During the follicular phase, under the
influence of oestrogens, the blood vessels in the submu-
cosa become hyperaemic, the endometrium becomes
oedematous, increasing its thickness, and the anechoic
vascular zone is more easily visualized (Griffin and
Ginther 1992). The content of the uterine lumen may
also vary during the oestrous cycle, being typically
absent during the luteal phase, whereas a small amount
of fluid may be present during the follicular phase
(Barlund et al. 2008). Nevertheless, the stage of the
oestrous cycle is better determined through the obser-
vation of the ovaries (Fig. 1).
Diagnosis of pathologic conditions of the uterus
Ultrasound examination of the uterus may help to
identify pathological conditions (Fig. 1). One of the
most frequent alterations that cause substantial eco-
nomic losses is uterine inflammation. In a recent study,
using cytological methods for the diagnosis, Gilbert
et al. (2005) found a prevalence of endometritis of 53%
of dairy herds in the USA. Using US and uterine
cytology, a study (Quintela et al. 2010) carried out in
north-west Spain indicated an incidence of endometritis
of 45.7%.
Uterine inflammations can be classified as metritis,
when they occur during the first 21 days after parturi-
tion, and local and general symptoms are present.
Endometritis beyond 21 days postpartum is character-
ized by local findings only (Sheldon et al. 2009).
Although postpartum metritis or clinical endometritis
can be detected through direct exploration of the
external genital organs or by rectal palpation and ⁄ or
vaginoscopy, ultrasonography is useful in the diagnosis
of uterine diseases (Barlund et al. 2008).
To determine the real usefulness of US for diagnosing
uterine infections, we are going to recall the common
methods used for an endometritis diagnosis. In several
studies (Kasimanickan et al. 2004; McDougall et al.
2007; Barlund et al. 2008) the results obtained when
using vaginoscopy, rectal palpation, US or a metricheck
have been compared with those obtained by uterine
cytology (considered the gold standard test for diagnos-
ing endometritis). All of them concluded that cytology
allows us to detect some endometritis that could not be
detected by any of the other methods, and that US
should be the second method of choice. Rectal palpation
and animal inspection is subjective, often fails to
account for normal events and variability in uterine
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Ultrasound in the Reproductive Management
Fig. 1. Ultrasound images of uterus of non gravid female. 1) Luteal phase; 2) Estrous phase; 3) Estrous phase; 4) Mucómetra; 5) Mucopurulent
endometritis; 6) Purulent endometritis1
involution (Lewis 1997) and is not useful for identifying
subclinical processes. Rectal palpation or the casual
observation of a genital flow is not sensitive enough for
diagnosing endometritis (Miller et al. 1980; De Kruif
et al. 1982). Vaginoscopy and Metricheck are not very
useful for diagnosing endometritis, as the absence of a
purulent vaginal discharge does not exclude the prob-
lem, and it also might lead to an erroneous diagnosis of
endometritis instead of vaginitis. Nevertheless, the
presence of purulent mucus in the vagina is still too
frequently used to diagnose endometritis (Sheldon and
Dobson 2004).
Kasimanickan et al. (2004), compared the use of
rectal palpation, US and cytology to diagnose endome-
tritis in 228 cows. They concluded that diagnoses based
on rectal palpation were not related to cow fertility,
whereas those based on the use of US or cytology
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showed a high correlation. Mateus et al. (2002) reported
that the amount of fluid detected in the uterine lumen by
US was positively correlated to the existence of bacterial
growth in the fluid samples. However, endometrial
cytology and US showed a low correlation, probably
because the two methods evaluate different parameters
(Kasimanickan et al. 2004); whereas the first one deter-
mines the cellular response and the second evaluates the
clearance mechanism of the uterus. The proportion of
endometritis not diagnosed by US varied between 10%
and 20%, depending on the use of a 5% or 10% cut-off
point for PMN in the cytology (Quintela et al. 2010).
Other uterine conditions that can be detected by US
include mucometra, pyometra, abscess of the uterine
wall. The ecographical difference between mucometra
and pyometra is the ecogenicity of the uterine content.
Whereas the sterile mucus has an anechoic appearance,
36 LA Quintela, M Barrio, AI Peña, JJ Becerra, J Cainzos, PG Herradón and C Dı́az
Fig. 2. Ultrasound images of ovary. 1) Dominant follicle; 2) Cohort of follicles; 3) Cavitary corpus luteum; 4) Non cavitary Corpus luteum;
5) Follicular cysts; 6) Luteinized cysts1
the purulent content shows a certain degree of ecoge-
nicity, depending on the amount of purulent material in
the mucus (Quintela et al. 2006). Abscesses in the
uterine wall will appear to have a hyperechoic content
surrounded by a wall of certain ecogenicity (Quintela
et al. 2009).
Exploration of the Ovaries
Physiological ovarian structures
Ultrasonographic examination of normal ovaries during
the oestrus cycle allows us to visualize two evolving
structures: the follicle and the corpus luteum. Ovarian
follicles have spherical shape, contain anechoic fluid and
can be found in variable numbers and size (Griffin and
Ginther 1992). The advantage of US compared with
rectal palpation is that the subjectivity inherent in the
estimation of size and content can be eliminated, as
the follicular fluid can be visualized and the diameter
can be measured. This makes it possible to detect the
presence of a cohort of follicles, dominant follicles or
preovulatory follicles (Fig. 2).
To determine the stage of the oestrous cycle of a cow,
however, the visualization of a corpus luteum (CL) and
estimation of its age is as important as the visualization
of the ovarian follicles. Besides milk or blood proges-
terone determination, ultrasound is believed to be the
most accurate method for detecting the presence of a CL
in a bovine ovary. Compared with rectal palpation, US
examination has been shown to be a more sensitive and
specific method: 95% sensitivity and 100% specificity vs
95% and 95.7%, respectively, for rectal palpation
(Ribadu et al. 1994); 84% sensitivity and 71% specificity
vs 79% and 40%, respectively, for rectal palpation
(Bicalho et al. 2008). Such differences are likely to be
because of the fact that many CL do not protrude
enough from the ovarian surface to be palpated, and
also because haemorrhagic and cavitary CL with a soft
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Ultrasound in the Reproductive Management
consistency may be confused with follicles (Hanzen
et al., 2000; Quintela et al. 2006). The incidence of
cavitary CL ranges from 25.2% to 78.8% during
dioestrus and decreases with the progression of the
oestrous cycle (Kahn 2010). In a study carried out on 28
cows, Perez-Marı́n (2009) found that 28.6% of the CL
were cavitary 4 days post-ovulation (p.o.) and 50% of
the CL were cavitary 8 days p.o., this proportion
decrease after that period. Gnemmi (2004) also indicated
that about 50% of the corpora lutea are cavitary during
the first 10 days of the oestrous cycle, about 30%
maintain the cavity between 10 and 13 days p.o., and
only a small percentage of the CL are cavitary during
pregnancy. It can therefore be assumed that when a CL
has a cavity, it is likely to be in the first half of the
oestrous cycle, and if there is no cavity, the CL is more
likely to be in the second half. The size of the CL may
also be used to estimate its age. Perez-Marı́n (2009)
indicated that the CL increases in size until about day 8
p.o., maintains a constant size until about day 16 p.o.
and decreases after this period.
Knowledge about the ovarian cycle and follicular
dynamics together with US examination of the uterus
and ovarian structures (number and size of follicles,
existence of a cavity and size of CL) allows us to
estimate the stage of the oestrous cycle of a cow. At this
point, new technologies, such as colour Doppler avail-
able in many portable US scanners, may provide new
and interesting information. By studying the blood flow
to follicles and CL, it is possible to determine with a
certain degree of accuracy the time of ovulation and of
CL regression (Herzog and Bollwein 2007).
Diagnosis of ovarian pathologies
Cystic ovarian follicles (COF) are an important cause of
subfertility in dairy cattle, as they extend the calving
interval (Fourichon et al. 2000). COF are follicles with a
diameter of at least 2 cm that are present on one or both
ovaries in the absence of any active luteal tissue and that
clearly interfere with normal ovarian cyclicity (Van-
holder et al. 2006). There are two types of COF
depending on the wall thickness: follicular cysts have a
wall thickness <3 mm and luteinized cysts have a wall
thickness >3 mm (Tebble et al. 2001; Vanholder et al.
2006). Ultrasound examination of ovarian cysts allows
us to distinguish follicular and luteinized cysts much
more efficiently than rectal palpation (Douthwaite and
Dobson 2000) (Fig. 2).
On average, the positive predictive values for follic-
ular or luteal cysts diagnosed by palpation are 66%,
whereas by using ultrasonography they are 74% for
follicular cysts and 85% for luteal cysts (Hanzen, et al.,
2000).
What it is really important is accurately diagnosing the
cystic structure. In a study on 584 cows (Stevenson 2006),
51 were diagnosed with ovarian cysts. Following a more
detailed follow-up, they concluded that only six were really
ovarian cysts. In 19 cows, the cyst coexisted with a CL, in 12
animals the cyst was suspected to be a preovulatory follicle
as it responded to GnRH treatment, and in 14 cows the cyst
was suspected to be a cavitary CL. Ultrasonography
allows us to reduce some of the confusing factors: the
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presence of a CL can be easily detected, the exploration of
the uterus may show an oedematous endometrium typical
of cows with a preovulatory follicle, but not normally seen
in cows with ovarian cysts (Bartolome et al. 2005; Arch-
bald et al. 2009), and the size of the follicular structure can
be objectively determined.
Traditionally, follicular cysts were treated with
GnRH, and luteinized cysts with PGF2a. However,
GnRH very often induced luteinization of the follicular
cyst, and a subsequent treatment with PGF2a was
needed to remove the luteinized structure. Nowadays,
combined treatments, such as Ovsynch or Ovsynch plus
a progesterone slow release device, have become com-
mon treatments for ovarian cysts (Fricke and Wiltbank
1999; Bartolome et al. 2000), and differentiation be-
tween follicular or luteinized cysts has no relevance
(Archbald et al. 2009).
Even though granulose-theca cell tumours are the
most common ovarian tumours in cattle, they are rare
(<0.5%). This type of tumour arises from the sex cord
stromal tissue within the ovary and may be relatively
small, solid, and yellow to white, or large, filled with
cysts of varying sizes and weight up to 11.9–12.3 kg
(Roberts 1986). The ultrasonographic appearance will
vary depending on the tumour aspect. An image
resembling a honeycomb is often seen because of the
presence of fluid-filled cavitary lesions inside the tumour
(Quintela et al. 2006).
Pregnancy Diagnosis
Early pregnancy diagnosis is particularly important in
dairy herds with a high proportion of undetected heats
(Walsh et al. 2011; Holman et al. 2011). Many non-
pregnant cows do not show heat, or the heat is not
detected by the farmer, which significantly increases the
days open (Roelofs et al. 2010). An early pregnancy
diagnosis facilitates the detection of non-pregnant cows
and enables them to be treated for a new insemination
(Romano et al. 2006) (Fig. 3).
For an early pregnancy diagnosis, the uterus should
be systematically explored. Exploring the whole uterus is
of the utmost importance, as during early pregnancy,
the majority of the endometrial surface is not different
from that seen in a non-pregnant cow. The embryonic
vesicle can be seen as an anechoic zone where the lumen
of the uterine horn is slightly dilated. Its appearance can
be confused with that of a uterus during oestrus or
endometritis. Inside the embryonic vesicle, the embry-
onic mass can be located (Griffin and Ginther 1992;
Quintela et al. 2006). Once the embryo is visualized, its
viability can be confirmed by checking the heartbeat
frequency that should be approximately 145 beats ⁄ min
between 26 and 60 days of pregnancy (Bo and Caccia
2000). If this systematic sequence is followed, the
diagnosis will be accurate. However, such accuracy will
also depend on the gestational age. Before 26–27 days of
pregnancy, using 5 MHz probes, about 40–50% of the
embryos cannot be visualized (Gnemmi 2004), as they
are adhered to the uterine wall at this time, and
consequently, the diagnosis cannot be accurate. After
27 days of pregnancy, the reliability of the diagnosis is
high, as shown in Table 1 (Quintela et al. 2006;
38 LA Quintela, M Barrio, AI Peña, JJ Becerra, J Cainzos, PG Herradón and C Dı́az

Fig. 3. Ultrasound images of pregnancy: 1) 29 days of pregnancy; 2) 34 days of pregnancy; 3) 44 days of pregnancy; 4) 51 days of pregnancy;
5) Twin pregnancy; 6) 75 days of pregnancy1

Table 1. Accuracy of ultrasound for pregnancy diagnosis, in function of pregnancy days

Days of pregnancy n Sensitivity (%) Specificity (%) Accuracy + (%) Accuracy ) (%) Total Accuracy (%)

24–26 205 100 36.36 84.4 100 85.71

27–29 752 99.72 86.28 92.1 99.5 94.6
30–35 1399 100 88.53 91.6 100 94.9
36–40 1029 99.83 89.15 92.9 99.7 95.4
41–45 918 100 89.64 94.3 100 96.2
46–50 756 100 84.65 93.3 100 95.1
51–60 866 100 84.34 94.1 100 95.5
>60 267 100 84.62 96.4 100 97.0
Total 6192 99.95 87.31 93.11 99.89 95.29

Romano et al. 2006). The accuracy of the diagnosis is
higher in negative than positive cases because of the
high incidence of embryo mortality in dairy cattle
during the second month of pregnancy, estimated at an
average of 12.8% between 27 and 51 days of pregnancy
(Santos et al. 2004). It accounts to some extent for the
false positives observed during early pregnancy diag-
noses.
US examination enables the detection of twins. The
presence of a higher amount of fluid in the uterine

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Ultrasound in the Reproductive Management
lumen, more alantoid membranes than expected for the
gestational age, together with the visualization of a
hyperechoic zone, which would correspond to the
anastomosis of alantoid membranes, and of two CL,
suggest the presence of a twin pregnancy (Quintela et al.
2009). The diagnosis of twins is important for dairy
farms as the risks of pregnancy losses, freemartinism or
dystocia are higher than in single pregnancies (Eddy
et al., 1991; Silva del Rio et al., 2009).In our recent study
(using personal data) on 802 twin pregnancies, the
proportion of gestation loss was 30.7%. This problem
has acquired more relevance during the last few years, as
it has been observed that as milk production increased the
percentage of twin pregnancies also increased. Lopez
et al. (2005) observed that in cows producing
<30 kg ⁄ day, the proportion of multiple ovulations was
0%, whereas in cows producing more than 30 kg ⁄ day,
about 51.6% of the cows had double ovulation. This
effect relates to a lower serum concentration of steroid
hormones, as a consequence of an increased blood flow to
the liver in high producing dairy cows (Wiltbank et al.
2000). In fact, in the 1980s the twinning rate was
Fig. 4. Ultrasound images of alterations of pregnancy: 1) Embryonic dead; 2) Fetus with ascites; 3) Fetus with umbilical cord dilatation; 4) Fetus
without ribs and lungs1
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approximately 1.5–2% (Kinsel et al. 1998), whereas in
the late 1990s it was around 4% (Silva del Rio et al. 2007).
Diagnosis of Alterations in Pregnancy
One of the most frequent alterations in pregnancy is
embryonic or foetal mortality. An embryo resorption
can be ultrasonographically recognized by the following
characteristics: the amount of alantoid fluid is lower
than expected for the gestational age; there is a presence
of membrane fragments freely floating in a cloudy fluid,
an absence of embryo ⁄ foetus, an absence of heartbeat.
The existence of one or several of these signs are
indicative of gestation loss (Gnemmi 2004; Quintela
et al. 2006) (Fig. 4).
Foetal malformations can be detected after approx-
imately 55–60 days of pregnancy. Most large organs of
the head, thorax, abdomen and pelvis were visible with
ultrasound during the first 4 months (Rajamahendran
et al. 1994). Alterations of the normal anatomy, as
for example, ascites, umbilical cord alterations, etc., are
indicative of reduced foetal viability, so it may be
40 LA Quintela, M Barrio, AI Peña, JJ Becerra, J Cainzos, PG Herradón and C Dı́az
appropriate to induce abortus to avoid increasing the
days open.
Foetal Sex Determination
Diagnosis of foetal sex could increase the price of a
pregnant cow, in which case it would be profitable. It
will also help dairy farmers in deciding whether or not to
retain pregnant cows already earmarked for culling,
depending on the sex of the foetus. The sexing of twins is
Fig. 5. Genital tubercle in 51 days bovine fetus (1). Ultrasound images
of 65 days male fetus (2) and 65 days female fetus (3)1
also advantageous as it allows selective termination of
unwanted pregnancies, such as those involving a male
and female foetus, which would otherwise bear a
freemartin (Rajamahendran et al. 1994).
The external genitalia are initially formed on the
caudoventral surface of the abdominal wall, between the
hind limbs (Des Côteaux et al. 2010). These primordial
structures are seen as poorly defined elevations and
constitute the genital tubercle (GT) (Fig. 5), the uro-
genital fold and the genital swellings, which are devel-
oped in the male or female embryos. In males, the GT
becomes elongated to form the penis, the urogenital fold
encloses the penis to form the prepuce and the genital
swellings become enlarged to develop into the scrotum.
In females, the GT forms the clitoris, the genital
swellings completely disappear and the urogenital folds
develop to enclose the GT, forming the labia (Noden
and De la Hunta 1985; Des Côteaux et al. 2010).
The GT is a reliable indicator for bovine foetal gender
determination between days 55 and 60 of gestation
(Curran et al. 1989; Shunichi et al. 1994). When the GT
is located immediately caudal to the abdominal attach-
ment of the umbilical cord, the foetus is male. On the
contrary, when the GT is located towards the base of the
tail, the foetus is female (Fig. 5) (Ali 2004). According to
Ali (2004), the best time for foetal gender determination
is between days 56 and 98 of gestation, although the
breed and age of animals should be taken in consider-
ation. Accuracy of early foetal sex determination by US
assessment of the relative location of the GT is high
when US examination is properly timed (Muller and
Wittkowski 1986; Curran et al. 1989; Wideman et al.
1989; Shunichi et al. 1994; Ali 2004; Quintela et al.
2011). The period of GT migration is a vital parameter
and must be verified to avoid male foetuses being
incorrectly classified as females because of GT migration
after examination (Santos et al. 2005).
Early studies reported that foetal gender could be
determined between days 73 and 120 post-insemination
by US visualization of the scrotum and mammary
glands of male and female foetuses (Muller and Witt-
kowski 1986). However, the possibility of gender deter-
mination decreases as pregnancy progresses after the
first trimester. In advanced pregnancy, two situations
occur that may hamper gender determination. Firstly, as
the foetus grows it becomes more difficult to move the
transducer relative to the foetus to attain the desired
ultrasound image. Secondly, the gravid horn is more
likely to have descended ventrally into the abdominal
cavity, so that imaging of the foetal gender is impossible
without retraction of the gravid horn (Beal et al. 1992).
Colour Doppler ultrasound
As described above, the black and white US has been
helping us to understand the reproductive physiology
and hence the development of reproduction manage-
ment systems in the cow. To gain more precise infor-
mation on the physiological status of the ovaries, the
local blood flow in individual ovarian follicles and CLs
can be observed using colour Doppler US (Herzog and
Bollwein 2007) This technique is based on the Doppler
effect (Johann Christian Doppler, 1903) that establishes
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Ultrasound in the Reproductive Management
that when an US bunch with a given frequency finds a
flow of erythrocytes, part of the US bunch is reflected
towards the probe with modified frequency, higher if the
interface is approaching to the probe and lower if it is
moving away. Therefore, by measuring the apparent
change of frequencies between emitter and receiver, the
blood flow velocity can be determined (Herzog and
Bollwein 2007).
A colour Doppler image shows the area where a
blood flow is detected, so the amount of colour
represents the volume of blood present in the examined
structures rather than its flow rate.
Current availability of portable high-quality colour
Doppler US scans has raised interest on the use of this
technology for reproductive management of cattle.
Nowadays, this technology is mainly used for research
purposes, but with the generation of new knowledge, in
the future, it may become a useful tool for the cattle
practitioner.
Blood flow changes in the ovaries
Colour Doppler US is a useful technique for evaluat-
ing ovarian vascular function, allowing a visual
observation of the blood flow in a delimited area in
the wall of preovulatory follicles (Brannstrom et al.,
1998) or within the corpus luteum (Miyazaki et al.,
1998; Acosta et al. 2002). This imaging has facilitated
haemodynamic studies and reproducible measurements
(Collins et al., 1991; Brannstrom et al., 1998; Acosta
et al. 2002).
The blood flow in follicles and CLs has a narrow
relationship with the growth, atresia and ovulation of
the follicle and with CL functionality.
Follicular growth: Acosta et al. (2005) found that a
high percentage of the small follicles with high blood
flow before deviation derived towards the second
large follicle or the largest follicle, and after deviation,
the blood flow was reduced in the second large follicle
and increased in the largest follicle. Before deviation,
the two types of follicles had a similar diameter, but it
was clearly different after deviation. The atretic
follicle was characterized by a lack of detectable
blood flow and a progressive decrease in diameter
(Acosta et al. 2003). These data suggest that follicular
growth, selection and atresia are closely associated
with changes in blood supply to an individual follicle.
Ovulation: Acosta et al. (2003) described changes in
the local blood flow in the outer teca layer of mature
follicles. Either the area of detectable blood flow in the
follicle wall and the time averaged maximum velocity
were small until the LH peak, when they increased and
kept high until ovulation. There is a narrow relation-
ship between the local blood flow in the follicular wall,
and the structural and functional changes occurring
during the ovulatory process.
Corpus luteum: The CL is one of the most highly
vascularized organs and receives the greatest rate of
blood flow per unit of tissue of any organ in the body
(Wiltbank et al. 1988). During its initial development
(48–120 h), a gradual increase in the blood flow
around the CL matches with an increase in its volume
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and in the serum progesterone concentration (Acosta
et al. 2003), which suggests that local blood flow in
the CL is related to its potential to produce and
release progesterone.
In the process of spontaneous luteolysis, blood flow in
the CL was observed to increase during the days 16–17
of the oestrous cycle, concomitant with an increase in
plasma levels of 14-dihydro-15-keto-prostaglandin F2a
(PGFM) and was followed by a decrease in plasma
progesterone concentration 1 day after (Shirasuna et al.
2008; Miyamoto et al. 2005; Shirasuna 2010).
Applications of Doppler Colour to Reproduction in Cattle
Functionality of the Corpus Luteum: Correct diag-
nosis of the presence of a CL and estimation of its
functional status are important in the reproductive
management of dairy cows. Luteal blood flow (LBF)
is more informative about luteal function during the
bovine oestrous cycle than is the assessment of luteal
size (LS) (Herzog and Bollwein 2007) In the luteal
growth phase LBF, LS and P4 increase simulta-
neously; during the luteal static phase, the three
parameters are high, however, during the regression
phase LBF and P4 decrease parallel, whereas LS
decreases later (Herzog et al. 2011). During the static
phase of the oestrous cycle and at the onset of CL
regression, the luteal size is similar and using only this
parameter may cause diagnosing errors.
Pregnancy diagnosis: Diagnosis of pregnancy via
transrectal palpation is possible as early as Day 35
after oestrus (Romano et al. 2007). When using B
mode US, to get a high accuracy, pregnancy diagnosis
is recommended to be carried out around day 28 after
AI, although the embryo might be seen a few days
earlier (Nation et al. 2003; Quintela et al. 2006). By
using colour Doppler US, pregnancy diagnosis can be
carried out earlier by evaluating the LBF, as non-
pregnant animals can be identified based on the
decrease in CL blood flow associated with luteolysis.
In pregnant cattle, active blood flow surrounding the
CL was continuously observed during the late luteal
stage (16–23 days after ovulation). However, in non-
pregnant cattle LBF clearly decreased at 19 days
after ovulation (Matsui and Miyamoto 2009; Utt
et al. 2009).
Ovarian cysts: Ultrasonographic differences between
follicular and luteinized cysts have been described
with B mode US, classifying the type of cysts
depending on the wall thickness. The use of colour
Doppler US helps to confirm the structure of the
follicular wall as it shows the location of blood vessels
in the wall thickness. Follicular cysts show the
presence of blood flow inside the wall, whereas
luteinized cysts show blood vessels surrounding the
wall (Matsui and Miyamoto 2009).
Embryo and foetal losses: An evaluation of the status
of CL blood flow could be used not only to diagnose
the pregnancy, but also early embryonic death.
Corpus luteum regression precedes embryo death
when it occurs before day 25, but if embryo mortality
occurs between days 25 and 40 of pregnancy, the CL
42 LA Quintela, M Barrio, AI Peña, JJ Becerra, J Cainzos, PG Herradón and C Dı́az

regression may delay up to 3 days (Kastelic et al.
1991). Thus, detection of a reduced LBF may predict
or confirm embryo mortality.
To the moment, studies in cattle indicated that LBF
was greater in pregnant than in non-pregnant cows
during dioestrus, but it was not a reliable method to
predict pregnancy loss in the individual cow, because
of substantial variation among cows (Herzog et al.
2011).
Blood flow to the follicle and fertility: In women, a
relationship has been established between the blood
flow in the preovulatory follicle and oocyte quality
(Palomba et al. 2006). Doppler ultrasonographic
characteristics of the preovulatory follicle 30 h after
hCG injection indicated that follicle blood flow was
greater in mares that became pregnant than in mares
that did not (Silva et al. 2006). In Holstein heifers
after treatment with PGF2a, GnRH after 36 h, and
AI 26 h after GnRH, Siddiqui et al. (2009) found that
blood flow at the time of AI was greater in the follicle
wall of the heifers that became pregnant than in
heifers that did not. It seems, therefore, that a direct
relationship exists between the blood flow in the
preovulatory follicle and the success of AI.
in dairy farms. Its high sensitivity and specificity for
detecting ovarian follicles, corpora lutea or cystic
structures, makes it useful for determining the stage of
the oestrous cycle of a cow or diagnosing ovarian
pathologies. Such information may be vital to make the
optimal treatment decisions in each particular case.
Ultrasound examination allows us to distinguish
between ovarian cysts and physiological structures.
Ultrasonographic examination of the bovine uterus
enables the detection of subclinical endometritis on a
farm.
Ultrasonography makes it possible to diagnose early
pregnancies with a higher level of accuracy than rectal
palpation. The detection of non-pregnant cows is of
utmost importance for reducing days open on dairy
farms, as one of the most critical problems in dairy
farms is inefficient heat detection. In addition, valuable
information on embryo viability, presence of twins,
normality of foetal development or foetal gender can be
obtained by US. Finally, colour Doppler ultrasound
can help to gain more precise information in determin-
ing the stage of the oestrous cycle of the cow, as well as
for early pregnancy diagnosis or diagnosis of embry-
onic loss.

Conflict of interest
Conclusions
None of the authors have any conflict of interest to declare.
Ultrasonography is a more accurate tool than rectal
palpation for reproductive control of cattle, particularly

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Submitted: 28 Dec 2012; Accepted: 3 Apr
2012
Author’s address (for correspondence): LA
Quintela, Faculty of Veterinary Medicine,
University of Santiago de Compostela,
27002 Lugo, Spain.
E-mail: luisangel.quintela@usc.es
All the figures are from our own work, and
have been taken with a portable US scan
Tringa linear, Esaote ⁄ Pie Medical, with a
transrectal linear probe of 5 MHz.
 2012 Blackwell Verlag GmbH