INTRODUCTION

In Veterinary field, ultrasonography has become an important diagnostic tool

for evaluating the female reproductive system. With the help of ultrasound
technology, it is possible to view the entire reproductive system in a non-invasive
manner (Carrière et al. 2002).
Ultrasonography is considered a wonderful tool particularly , for the study of
reproductive functions viz. follicular dynamics, pregnancy diagnosis and
abnormalities of contents of tubular tract. In 1980, Palmer and Driancourt reported for
the first time, real time ultrasonography for diagnosis of pregnancy in the mare. This
report generated considerable interest among veterinarians and scientists about
ultrasonography and this new discovery contributed significantly towards
understanding of functioning of the uterus and ovaries of mare, cows and other
species. The ultrasonography is fast replacing the age-old procedure of rectal
palpation of genital tract for pregnancy diagnosis. This technique has been used and
advocated for the use in the academics and field practice.

Development of diagnostic ultrasound

Ultrasound is defined as any sound frequency above the normal hearing range
of the human ear; i.e. greater than 20,000 Hz. Sound waves in ultrasound devices are
typically produced by vibrations of specialised crystals (piezoelectric crystals) housed
in an ultrasound transducer, with the vibrations of the crystals themselves produced by
pulses of electric current. A proportion of the sound waves reflected back to the
transducer is converted to electric current and displayed as an echo on the ultrasound
viewing screen. The transducer, therefore, acts as both the sender and receiver of
echoes. The echoes are evident on the viewing screen as varying shades of gray (black
to white). Early applications of ultrasound as a diagnostic aid in medicine utilised
Amplitude or A-mode ultrasound. The first use of ultrasound as a diagnostic aid in
veterinary medicine was for the detection of pregnancy in sheep. Now a days,
Brightness (B) mode and Doppler are more commonly used than A-mode, and a
variety of applications have emerged using these techniques. The introduction of
computer systems to ultrasound machines has enabled the storage, processing and
presenting of large amounts of data, allowing the production of static two-dimensional
grey scale images and real-time imaging]. This real-time B-mode imaging is
currently the form of ultrasound most commonly used.

ULTRASONOGRAPHY MACHINE FOR VETERINARY APPLICATION

The application of real-time ultrasonography to study animal reproduction
represents a technological breakthrough that has revolutionised knowledge of
reproductive biology. New information generated through ultrasonic imaging has

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clarified the nature of complex reproductive processes in animals, including ovarian
follicular dynamics, corpus luteum function, and foetal development. Until recently,
the techniques used in studying patterns of follicular development involved the
measurement, counting and histological evaluation of ovaries of animals killed at
various stages during the oestrous cycle, or marking of follicles with ink, followed by
serial laparoscopy. In contrast, the development of ultrasonic probes that can be used
intrarectally to visualise ovaries has opened up new possibilities for examining the
dynamics of follicular growth and regression and provided a means for repeated,
direct, noninvasive monitoring and measuring of follicles within the ovary. Early
utilisation of ultrasound technology in the dairy industry has included applications
such as transvaginal follicular aspiration and oocyte recovery, and as a complementary
technology for embryo transfer procedures. In addition, early pregnancy diagnosis and
monitoring of foetal viability is a great advantage of ultrasonography.
Trans-rectal real-time B-mode ultrasonography is being routinely used as
research and diagnostic tool in bovine reproduction. Real-time ultrasonic imaging
provides a noninvasive technique to image directly, in situ, the internal and external
anatomy of reproductive organs and tissues, and to characterize reproductive events
(e.g., ovulation, transition of the uterus from a diestrous to an estrous echotexture).
The technique is noninvasive, relatively simple and effective. By using real-time
ultrasonography, it is possible to examine the individual ovary and follicular dynamics
on a daily basis. Ultrasonographic images reveal that the growth of follicles in bovine
ovaries occurs in a wave-like manner with two or three waves per estrous cycle.
Different types of structure develop on the ovary like corpora lutea, follicles, follicular
and luteal cysts and ovarian tumors, which are difficult to diagnose by manual per-
rectal palpation. These can easily and accurately be diagnosed by using
ultrasonography. To improve the reproductive performance of cow, it is necessary to
inseminate at perfect time to improve the chance of fertilization. For this, it is
necessary to correctly assess the onset of estrus and time of ovulation which can be
accomplished by rectal ultrasonography. Early pregnancy diagnosis, fetometry to
estimate fetal age and assessment of progression of fetal growth, diagnosis of
pregnancy disorders and fetal sexing can be accurately done by using ultrasonography.
The course of uterine involution and uterine disorders (endometritis, pyometra,
hydrometra) can be monitored by ultrasonography (Kahn and Ludlow, 1989).
The user needs to master the physical principles of ultrasonography in order to
perform a worthwhile examination with the instrument. The quality of the images
depends above all on the user's understanding of the interactions between the
ultrasound wave and the organ tissue, as well as proper use of the instrument's
controls (DesCôteaux et al. 2005).

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BASIC PRINCIPLES, PROBE INDICATIONS AND TERMINOLOGY

Probe characteristics and resolutions:

The probe is the most fragile component of the ultrasound apparatus. In the
veterinary market, usual probes have a frequency of 3.5, 5.0 or 7.5 MHz. The ability
of the instrument to distinguish between two structures located very close together
along the axis of the ultrasound beam is called its axial resolution (Figure 1). The axial
resolution is best when the groups of waves emitted have a short wavelength. Since
the number of cycles in each group of waves is set according to instrument design, the
only way to shorten their length is to use a probe with a higher frequency. For
example, the group of waves produced by a 7.5 MHz probe will be shorter than one
produced by a 3.5 MHz probe and will provide better axial resolution.
There are two types of probes: linear and sectorial (Figure 2). In
theriogenology, linear probes are preferred for transrectal ultrasound examinations of
the ovaries and uterus. This probe has a set row of crystals that are selected
electronically to form a rectangular image. The linear probe provides good resolution
for tissues located close to the probe. Sectorial probes have one or several crystals
whose position produces a beam in the shape of a pie slice. The advantage of the
sectorial probe is that it doesn't require a large surface of contact, and it scans a greater
overall surface. The disadvantage is that the visual field and the lateral resolution (i.e.
the ability of a system to differentiate between two adjacent structures) are more
restricted close to the probe. The sectorial probe is ideal for viewing the small
ruminant fetus by transabdominal ultrasound imaging and for ultrasound-guided
transvaginal aspiration of bovine follicles.

Figure 1.Axial and lateral resolutions of the linear probe

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 Figure 2.Types of probes (A = sectorial and B = linear)

The details contained in an ultrasound image (resolution) as well as the depth of the
tissue observed depend on the frequency and the focalization of the scanning beam.
With a lower frequency, tissue penetration will be deep, but the resolution will be
lower. A higher frequency enables better resolution, but beam attenuation will be
greater and it will not penetrate the tissue as deeply. Table I summarizes the
indications of the different types of probes used in bovine theriogenology.

Table I.
Summary of indications and characteristics of different types of probes used in
bovine Reproduction

Terminology and interpretation of images

The description of ultrasound images is based on an evaluation of the shape,
contour, size, and position of the structure being studied, as well as its echogenicity,
which depends on the amplitude of the echoes received. An echogenic structure
reflects the majority of soundwaves back to the probe and thus appears from white to
different shades of grey on the screen. An anechogenic structure does not produce
echoes; instead, it transmits the waves on to more deeply situated tissues.

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 An example of an anechogenic structure is follicular fluid, which appears
black on the screen. The terms hypoechogenic and hyperechogenic indicate
respectively a decrease and an increase in relative echogenicity in comparison with the
surrounding tissue, whereas the term isoechogenic is used to describe similar
echogenicity with the surrounding tissue.

The following sections will present the principal uses of ultrasonography in bovine
reproduction.

Ultrasonic imaging
The use of ultrasound as a diagnostic technique has made an important
contribution to the understanding of reproductive problems in farm animals. The
earlier methods such as those based on the Doppler principle have now been
superseded by real-time ultrasound machines with two-dimensional on-screen images.
The most practical type of instrument for use in cattle and buffalo is one based on
real-time B-mode ultrasound and equipped with a linear-array 5 or 7.5 MHz intra-
rectal probe.
The method is non-invasive and interactive, it can be used for imaging the
uterus, ovaries and related structures. With experience, pregnancy can be diagnosed as
early as 10 to 15 days after mating. However, some of those conceived will not
survive, due to the natural occurrence of embryonic deaths in about 20 per cent of
conceptions. In practice it is more common to perform diagnoses at 20 to 30 days after
mating, when the embryo and its heartbeat can be discerned. At later stages, the age
can also be deduced from the heart rate and length of the embryo.

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 The value of the technique, as with progesterone assay, is in detecting non-
pregnant cows by 18 to 20 days, so that they can be observed more closely for a return
to heat and mated at the correct time. The method is now being used increasingly by
veterinarians in industrialized countries. The main limitations to its application in
developing countries at present, are the capital cost of equipment and the dearth of
veterinarians experienced in the method. However, there are indications that simpler,
more portable and possibly cheaper instruments will be developed in the future. If so,
this technique could become a valuable tool for veterinarians servicing smallholder
farmers.

Type of ultrasound machines

In Indian market many kinds of ultrasound machines are available. Many
institutions are using a machine”Vet scanner 200” from Pie-medical (The
Netherlands). Other machines used are from Toshiba (Japan), Aloka (Japan), Siemen
(Germany), Philips (USA), Madison (South Korea).

Types of ultrasonography:
In veterinary practice, four types of ultrasonography are in use viz. A- mode,
B-mode, M-mode and Doppler ultrasound.
A-Mode:
This kind of ultrasonography is used to know distance of a particular point in
the form of amplitude of returned sound beam. This is also known as amplitude
modality and is one-dimensional display of echo amplitude as per the distance or time.
Generally, it is displayed in the form of graph, which provides distance on horizontal
axis and amplitude on the vertical axis.
B-mode:
It is brightness mode, which provides two-dimensional image of the
tissue/organ scanned. It is most commonly used in veterinary practice as it provides
graphic image on the screen. Brightness of the tissue/organ is in proportion to the
amplitude of the reflected echoes returning to the transducer. Solid structures appear
bright (white), while fluid appears black. The image appears live due to continuous
updating of the image. Therefore, it is also termed as real time ultrasonography.
M-mode:
It is the motion modality, which provides tracing on the screen that can be
recorded on a thermal paper. It is used to scan heartbeats or moving structures/tissues.
It provides one-dimensional image on vertical axis versus time on the horizontal axis.
Doppler ultrasound:
It also works on ultrasound-beam reflections and is used for moving objects. If
the motion of the moving object is towards the transducer, then the frequency of the
sound beam is increased. If the motion is away from the transducer, then the

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frequency of the sound beam is decreased. It is of two kinds: 1) Continuous-wave
Doppler system and 2) Pulse-wave Doppler system. In continuous-wave Doppler
system, there are separate transmitting and receiving crystals, while in pulse Doppler
there is a single crystal for doing both the jobs (transmission and reception).
Doppler ultrasound is, generally, used for haemo-dynamics and is also used for
pregnancy diagnosis in pigs. It has its usefulness in advanced stages of pregnancy to
detect fetal pulse When an ultrasound beam encounters a moving object such as a red
blood cell in vascular flow, the frequency of the returning echo is altered. An increase
in frequency occurs when the object is travelling towards the transducer; this is known
as positive Doppler shift. An object travelling away results in reduced frequency and a
negative Doppler shift. The measurement of these alterations in the returning echo
allows the direction and velocity of the flow encountered to be determined.With this,
colour flow Doppler is used to screen uterine and testicular blood flow and in the
diagnosis of the ovarian cysts in cattle.

General description of the images

It is important to realize that the image on the ultrasound screen represents a
fine section of an organ, grossly resembling a weakly-magnified histological cut. The
probe therefore simulates the passage of a knife, slicing through an organ tissue from
top to bottom. Ultrasonography thus presents a flattened two-dimensional image of a
finely-cut section of tissue; whereas radiography is a two-dimensional superimposed
view of the entire thickness of an animal or of a limb under observation.
Ultrasound images are rapidly renewed and images are superimposed one over
the other as the probe moves across a tissue surface. The rapid succession of tissue
section views gives the impression that the structures are moving like an animated
cartoon. When interpreting sectional views of an organ on the screen, it is essential to
have a good appreciation of the three-dimensional shape of the organ in space.

Techniques of examination
For transrectal or transcutaneous ultrasound scanning in animals, no sedation
is required, as the procedure is totally non-invasive and well tolerated. Adequate
restraint is required and the scanner should be placed at a sensible distance from the
animal on the side opposite the operator’s rectalling arm. All precautions that apply
to palpation per rectum are applicable to transrectal scanning. All faeces from the
rectum should be evacuated prior to introduction of the transducer. It is often
advantageous to carry out a preliminary exploration of the topography of the
reproductive tract before commencing the ultrasonographic examination. The
transducer face must be lubricated with a suitable coupling medium and is usually
covered with a lubricated plastic sleeve before insertion in a cupped, lubricated hand
through the anal opening in large animals or by using a rod stick in small animals. It
is then progressed cranially along the rectal floor to overlie the reproductive tract.

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The transducer face must be pressed firmly against the rectal mucosa in order to
effect ultrasound transmission through the rectal wall into the abdominal viscera.
The probe is moved across the reproductive tract in a thorough and systemic manner.

Images of reproductive tract:

Ginther and his graduate and postgraduate associates (at Wisconsin, USA)
have been engaged in ultrasonographic imaging of reproductive tracts of farm
animals. There are many other scientists, who are employing this tool for diagnosis of
reproductive problems. In fact, there are many areas, where imaging through
ultrasonography has stepped in as an crucial tool.

Imaging the Bovine Ovary: -

A thorough reproductive examination in cattle includes visualization of the
major structures on both ovaries. Although rectal palpation can be an accurate method
for diagnosing pregnancy, rectal palpation is a poor method for resolving ovarian
follicles (Pieterse et al., 1990b). By contrast, ultrasonic imaging is a highly accurate
and rapid method for assessing ovarian structures.

(1) Ovarian follicles

(2) Anovulatory follicles
(3) Corpus Luteum
(4) Luteal cysts and Follicular cysts

Ultrasonographic Image of Ovary; there are many black circles inside the ovary. These
circles are “Follicles”. (Because the ultrasonic wave can pass through the fluid, any fluid-
containing parts look black. Meanwhile hard tissues such as bone look white.). Also we can
know the exact size of the organ by left-side scale or a measuring function usually available
in most of the machine.

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(1) Ovarian Follicles (Folliculogenesis): -
Folliculogenesis is the process of forming mature follicles capable of ovulation
from the pool of nongrowing, primordial follicles in the ovary (Spicer and
Echternkamp., 1986). Ovarian follicles are fluid-filled structures surrounded by an
inner layer of granulosa cells and an outer layer of theca cells. The oocyte is suspended
within the antrum by a specialized pedicle of granulosa cells, called the cumulus
oophorous. Because fluid absorbs rather than reflects ultrasound waves, fluid-filled
structures such as antral follicles appear as black circular structures surrounded by
echogenic ovarian tissue that are spherical to irregular in shape.
Most veterinary grade ultrasound scanners can resolve ovarian follicles with a
diameter of 2 to 3 mm or greater, and larger antral follicles can easily be tracked
during serial scanning sessions (Pierson and Ginther, 1988).
Ultrasonic monitoring of individually identified follicles has been found to be a
powerful tool for studying follicular dynamics during estrous cycle and pregnancy,
and the intraovarian relationship among dominant and subordinate follicles.
Through ultrasonography it is now established that follicular growth occurs in
wave-like patterns during each estrous cycle. It has been shown that follicular growth
and regression continue during early pregnancy, as well as in the postpartum anestrous
period. Ultrasound has also helped us to understand the influence of dominant follicles
on medium and small follicles.

(2) Diagnosis Of Anovulatory Follicles: -

During estrus, the ultrasonographic image of anovulatory follicles is almost
similar as that of normal follicle. If a follicle has luteinized, then the image will be a
mixture of fluid (black) and white echoic reflections in the fluid that indicate
luteinization of follicle (Ginther and Pierson, 1984b; Griffin and Ginther 1991).
These white hyperechoic images within follicular fluid are seen generally when mare
is out of estrus, which should definitely be suspected for luteinization of anovulatory
follicles. This kind of image is seen in early diestrum, later on the image appear
similar to corpus luteum. Now, computerized programs are available to score intensity
of follicular fluid that can help in diagnosis of normal and abnormal follicles.

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 Ultrasound image of bovine ovaries prior to emergence of a follicular wave
(note two small follicles [< 5 mm]; Panel A), during proestrus (note preovulatory follicle
[13 mm]; Panel B), and after development of a follicular cyst (note delamination of
granulose layer into the antrum; Panel C). Images were taken using a 7.5 Mhz
transducer (Lamb, 2001).

(3)Corpora Lutea: -
The CL is a transient endocrine gland that forms after ovulation from the
tissues that previously constituted the ovarian follicle. The ultrasonic characteristics of
corpus luteum and luteal dynamics have been studied in heifers, mares, and lIamas.
Corpus luteum appears as distinctly echogenic area within the ovarian stroma. Two
types of luteal morphology have been described in cow; corpus luteum with and
without a fluid-filled central cavity. The central cavities appear ultrasonically as
echogenic (black) to hypoechogenic areas surrounded by relatively hyperechogenic
luteal tissue.

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 Ultrasound image of bovine ovary with corpus luteum(CL)and follicle(F)

Table: Ultrasonographic classification of CLs based on appearance compared with

plasma progesterone concentrations:

Ultrasonographic n
Plasma progesterone concentration
CL classification
<1 ng/ml 1-4ng/ml >4ng/ml
n % n % n %
CL not detected 30 21 70 9 30 0 0
Evolving CL 25 5 20 16 64 4 16
Mid-cycle CL 44 1 2.3 4 9.1 39 88
.6

Table: Ultrasonographic detection of functional CLs based on appearance compared

with plasma progesterone concentrations:

Ultrasonographic CL n Plasma progesterone concentration

classification
<1 ng/ml 1 ng/ml
N % n %
CL not detected 30 21 70 9 30
CL detected 69 6 8.7 63 91.3

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 Table: Ultrasonographic classification of CLs based on appearance compared with
plasma progesterone concentrations:

Ultrasonographic CL n Plasma progesterone concentration

classification

4 ng/ml >4 ng/ml

n % n %
Evolving CL 25 21 84 4 16
Mid-cycle CL 44 5 11.4 39 88.6

The close relationship between luteal size and progesterone concentration in some
species indicate that ultrasonic assessment of the corpus luteum is a valuable adjunct to
Plasma progesterone assay as an index of luteal reaction (Kastelic et al., 1990a).

Diagnosis of luteal and follicular ovarian cysts by per-rectum and linear-array

ultrasonography in dairy cows: -
Ultrasonograms of cysts diagnosed as luteal had grey echogenic patches along
the inner cyst wall and within the antrum of cyst. In contrast, ultrasonograms of
follicular cysts had an anechogenic antrum and a smooth, thin wall with few or no
grey patches.
Sensitivity and specificity of palpation per-rectum for diagnosis of type of
ovarian cysts were low 43.3 and 64.7%, respectively. In contrast, sensitivity and
specificity of ultrasonography were considerably higher 86.7 and 82.3 %, respectively.

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Table-: Agreement between palpation per rectum and linear-array ultrasonography for
diagnosis of luteal and follicular cysts in cows.

Type of cyst diagnosis No No. (%) correct diagnosis relative to

by method . progesterone classification*
Palpation Ultrasonogra Palpatio Ultrasonagraph Palpation and
phy n y Ultrasonagraph
y
Luteal Luteal 14 - - 12 (85.7)
Follicular Follicular 13 - - 10 (76.9)
Luteal Follicular 5 1 (20.0) 4 (80.0) -
Follicular Luteal 15 1 (6.7) 14 (93.3) -
Agreement between methods (14+ 13)/47x100=57.4%

Kappa coefficient = 0.2

* Luteal cyst (>0.5 ng/ml); follicular cyst (0.5 ng/ml).

Table: Agreement between clinician A and clinician B when diagnosing luteal and
follicular cysts by linear-array ultrasonography.

Type of cyst diagnosis by n No. (%) correct diagnosis relative to

clinicians progesterone classification*
Clinician A Clinician Clinician Clinician B Clinician A and
B A Clinician B
Luteal Luteal 28 - - 25 (89.3)
Follicular Follicular 12 - - 11 (91.7)
Luteal Follicular 1 1 (100.0) 0 (0.0) -
Follicular Luteal 6 3 (50.0) 3 (50.0) -
Agreement between methods (28+ 12)/47x100=85.1%

Kappa coefficient = 0.7

These data suggest that linear-array ultrasonography is more effective than palpation
per-rectum for diagnosing type of ovarian cyst in cow.
A definite diagnosis of luteal cysts with ultrasonography and subsequent
treatment with Prostaglandin F2 (PGF2) results in shorter interval to first estrus
after treatment or treatment of cysts either GnRH alone or on injection of PGF2, 9
days after GnRH treatment, but former treatment is better. This management practice
of ultrasonography enhances the reproductive efficiency.

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Imaging The Bovine Uterus: -
In cow (Pierson and Ginther, 1988) uterine echo texture was characteristically
dark during the period corresponding to follicular dominance (estrus), reflecting an
extensive degree of endometrial edema.
Trans-rectal ultrasonography has permitted assessment of changes in the shape
of the uterus during various reproductive states in the heifer (Pierson and Ginther,
1988). under luteal dominance, the uterine horns of cows were maximally curled,
during follicular dominance the uterine horns were less curled in cattle.
It was concluded that ultrasonography is a valid and repeatable technique for
quantifying uterine activity (Griffin and Ginther, 1992). Thus ultrasonography
provides a tool for immediate sequential assessment of uterine contractibility without
invading the uterus or attaching or inserting monitoring devices (e.g., electrodes). This
technique also has been used to assess the role of ovarian steroid on uterine activity.
Uterine cysts are ultrasonically visible as immobile, usually compartmentalized, non-
echogenic structures with well defined borders (Ginther and Pierson, 1984a).
Ultrasonic imaging has provided a much needed for continuing assessment of
uterine cysts without interfering with their development. Fetal remnants (e.g. fetal
bones, mummified fetuses) are ultrasonically detectable as brightly echogenic
structures with the uterine lumen (Ginther and Pierson, 1984a). Many smaller bone
remnants (e.g. 6 mm) are difficult to detect with trans-rectal palpation, and their
identification and removal is greatly facilitated by ultrasonography. The ultrasonic
appearance of some bovine uterine abnormalities has been described. Clearly,
ultrasonic imaging is an effective tool for diagnosing and uterine pathology and
classify its relationship to fertility and for evaluating treatment regimens.

Uterine Pathology: -
The use of ultrasound for post-partum or post-estrual examination will allow early
detection of potential uterine pathology. Ultrasound will allow differentiation of
(a) Uterine cysts (Focal fluid filled areas)
(b) Pyometra (Fluid filled with increased echogenisity)
(c) Dead fetus (No heart beats)
(d) Resorbing fetus (Poorly delineated gestational sacks with or with without fetal
remains)
(e) Premature placental separation (fluid between uterine wall and placental
tissue).

Endometritis / other fluid inside the reproductive tract: -

Ultrasonography is being used for differential diagnosis of endometritis from
other kind of fluid in the tubular tract. The diagnosis of endometritis is based on
echogenicity of fluid in the uterus. Normal estrus secretion appears anechoic. The
fluid is also observed in postpartum uterus for few days of parturition. The
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echogenicity of lochia deceases slowly over first few weeks. Immediate after
parturition, the cellular component of lochia may give echoic reflections. The outline
of fluid accumulation in the endometritis can be easily identified (Kahn, 1994). The
secretion/fluid in endometritis is comparatively echoic as it contains debris due to
cellular damage. However, the degree of change varies from occasion floccular echoic
spotted bright intensity in comparison to surrounding uterine wall. As in most of
endometritis cases, the organisms are anaerobic which produces bubbles that in turn to
produce floccular reflections. Adam’s et al., (1987), suggested that endometritis could
be diagnosed in a better way from mid to late diestrum phase.

Sonogram of Endometritis

Pyometra: -
Pyometra needs differentiation from endometritis and an experienced operator
can do it easily as the secretion of pyometra cases is thick and the ultrasonic images
show dilation of uterus, while it is not so in endometritis. The pyometra cases can be
differentially diagnosed with more echoic spot in the fluid and echogenicity increases
as the probe/transducer is moved ventrally i.e. base/bottom of uterus, which is due to
more sediment on the ventral aspect of uterus. Operator must differentiated image of
high viscosity urine in the bladder, which can be confused with a slightly cranially
dilated uterus. It is suggested that the pyometra should only be diagnosed if two
completely separate closely opposed hollow organ can be demonstrated. It is also
advised to differentiate amniotic fluid of later stages of gestation with echoic image of
pyometra. One should look for floating images of amniotic membrane in normal
pregnancy (Ginther, and Pierson, 1984a).

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 Sonogram of bovine uterus with pyometra

Ovarian tumors:
Ultrasonic images of granulosa cell tumor appear like a superovulatory ovary
with multiple follicles; however more vesicular structures are counted in this type of
tumors than normal ovarian follicles. In an ovary, few follicles (up to 5 or10)can be
seen at one time, while in granulose cell tumors more number of vesicles of higher
diameter (more than 10 mm) is seen in ultrasonographic image. Occasionally one
large sized vesicle is seen in granulose cell tumor that will be of a size that can not be
normal follicle (more than 50 mm), while in other cases many small vesicles, closely
associated can be seen. Other kind of tumors is less frequently scanned so far.

Granulosa cell tumor

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Transrectal ultrasonographic findings in various reproductive problems

Type of case Ultrasonographic finding

Endometrits Moderate to large amount of anechoic fluid in lumen alongth with

"snowy" echogenic particles. Uterine wall is thickened.
Uterine wall is thickened and uterine body is distended. Enormous
Post-partum
amount of anechoic fluid is present in the lumen along with with
Metritis
echogenic particles.
Uterus distended and have thickened uterine wall. The viscous
Pyometra fluid inside the lumen contained diffused, echogenic particles,
floating in it. Sonographic image resembles with uterine tissue.
Uterus is thin walled which is visible as white streak and appears
Mucometra to be filled with echogenic particles in the enormous anechoic
fluid present inside it.
Follicular Imaged as thin walled (≤2mm) non-echogenic structures visible
cyst over the surface of the ovary. Size is ≥25 mm

Anechoic central cavity bordered by a distinct wall (2-5 mm) of

Luteal cyst
luteinized tissue imaged over the ovarian surface. Size >25 mm.

Hypoechogenic membrane loosed their tense appearance and

begins to appear as floating with in the anechoic uterine lumen.
The conceptus, with membranes may be imaged in any area of
Abortion
birth canal accompanied by anechoic fluid. CL generally
maintains good size and echogenicity for a number of days post
abortion.
Uterine wall were thickened and bone fragments were seen as
dense echogenic areas shadowing the tissue below. Surrounding
Macerated
and between the bones purulent fluid was present with its typical
fetus
appearance of a non echogenic background containing many
small echogenic particles.
Uterine fluids were not evident in contrast to the snowy
Mummified appearance of the lumen of uteri in which macerated foetus were
foetus seen. Uterine wall is thickened. Fetal parts are rarely identified
and mostly appeared as a poorly defined echogenic mass.
Medium or thick band of hypoechoic streak is observed along the
Adhesions
uterine wall.
In 2 of the cases CL was imaged as distinctly echogenic areas
with in the ovarian stroma while in rest of the cases, on
Anestrus sonographic scanning, no echogenic area was imaged in the
ovarian stroma and small follicles (2-5 mm diameter) was visible
on the ovarian surface.
Ovarian A pus filled cavity is observed over the ovary imaged as fine dots
abscess of echogenic particles in the anechoic fluid matrix.

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Early Embryonic Loss:
Embryonic death, as indicated by cessation of heart beats. The embryonic mass
increased in echogenicity and was maintained with a gradually decreasing fluid
volume until the occurrence of estrus 17 days after death. Pregnancy loss contributes
to reproductive inefficiency because fertility assessed at any point during pregnancy is
a function of both conception rate and pregnancy loss. Conception rates at 28 to 32 d
post AI in lactating dairy cows range from 40 to 47% (Pursley et al., 1997; Fricke et
al., 1998) whereas, conception rates in dairy heifers are nearly 75% (Pursley et al.,
1997). Similarly, pregnancy loss in lactating dairy cows is greater than that in dairy
heifers (20 vs. 5%; Smith and Stevenson.,1995). Although the specific factors
responsible for early embryonic loss in dairy cows are not known, they may be similar
to those factors responsible for reduced conception rates.
Of cows diagnosed pregnant at 28 d post AI, 10 to 16% experience early
embryonic loss by 56 d post AI (Mee et al.1994; Vasconcelos et al.; 1997; Fricke et
al., 1998). Although the rate of pregnancy loss is significant in studies using
ultrasound to assess the rate of loss, the technique of ultrasound itself has not been
implicated as a cause of embryonic death in cattle (Ball and Logue., 1994.; Baxter and
Ward., 1997). Furthermore, ultrasound is a much less invasive technique for early
pregnancy diagnosis than is rectal palpation (Paisley et al., 1978; Vaillancourt et al.,
1979) and may minimize the rare incidence of palpation-induced abortions.

Evaluation of Artificial Insemination or Embryo Transfer Technique

Proper placement of semen or an embryo in the female tract is essential for
achieving high pregnancy rates when cattle are bred by artificial insemination or
embryo transfer. Transrectal ultrasonography can be used to identify the site of semen
or embryo deposition. Correct placement of semen or an embryo is performed by
using a metal bead which deposited semen or an embryo in the reproductive tract
(Beal et al., 1989). The position and direction of the highly-echogenic bead is guided
by ultrasonographic scanner. Ultrasound evaluation of the technique used by
inseminators or embryo transfer technicians allows the correct deposition of semen or
embryo.

RECENT ADVANCES IN ULTRASOUND

Advances in hardware and software

Recent advances in diagnostic ultrasound have resulted from concurrent
developments in the computer industry and a reduction in the size of component parts,
significantly influencing equipment design. Ultrasound machines are currently
available in a huge range of sizes. Battery operated hand-carried ultrasound scanners
are available]. Although initially intended for use in small animals, this type of
equipment is now increasingly being used in conservation projects for the
reproductive management of farm, wild and captive endangered species including
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elephants] and rhinoceros]. All of these machines are capable of B and M-mode real-
time imag-ing and many now also incorporate colour flow Doppler, increasing the
scope of the examinations that can be performed. All ultrasound machines allow
individual images to be captured and displayed. Combining computer technology with
medical ultrasound can help with displaying the data in a more appropriate fashion.
With this, advanced post-processing functions have given the operator greater ability
to optimise image quality, therefore allowing the production of vastly superior images
and Doppler traces. For example, three-dimensional ultrasound has been used in
horses to examine the reproductive tract]. Moreover, four-dimensional ultrasound,
where three-dimensional images are viewed in real-time, is now available.

Future Perspectives: -
(1) Colour Doppler technique: Vascular system of reproductive system can be
visualized in colour.
(2) Coloured USG – Better resolution & contrast.
(3) 3-Dimensional USG images are now available.

Diagnostic Limitations Of Ultrasonic Imaging: -

(1) Practically single ultrasound examination conducted at a given point in time as
opposed to serial ultrasound examinations.
(2) The physiological status of a follicle (e.g., dominant, subordinate, growing,
regressing) or corpus luteum cannot be determined during a single ultrasound
examination.
(3) Ultrasonic imaging aids in distinguishing anatomical attributes of a structure but
differ in little information regarding physiological or endocrine status.
(4) Practically single ultrasound examination conducted at a given point in time as
opposed to serial ultrasound examinations.
(5) The physiological status of a follicle (e.g., dominant, subordinate, growing,
regressing) or corpus luteum cannot be determined during a single ultrasound
examination.
(6) Ultrasonic imaging aids in distinguishing anatomical attributes of a structure but
differ in little information regarding physiological or endocrine status.

Conclusions:

The impact of real-time ultrasound on the study of animal reproduction has been
dramatic, and development of portable ultrasound machines has given clinicians an
added tool for diagnostic reproductive management. Ultrasound is commonly used to
monitor uterine anatomy, involution and pathology. In addition, it has been used to
detect pregnancy, study embryonic mortality, monitor foetal development, and
determine foetal sex. Recent advances in ultrasound technology in both hardware and
software have resulted in the production of superior images and the widespread use of
ultrasound. Another advantage of ultrasound is its real-time nature in examination,
allowing studies of moving structures.
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The procedure of ultrasonography is simple, but one needs thorough knowledge of
machine and location of desired organs. It takes a while to obtain good quality images.
For an experienced monographer, it is most convenient procedure. Unlike, X-rays, it
has no hazardous effect on the operator. So far no major undesired effects are
observed on the animal tissues/organs. It provides quick and reliable results. The B-
mode ultrasonography has proved a boon for veterinary practice of farm and pet
animals. Therefore, ultrasonography is becoming indispensable tool for diagnosis of
reproductive disorders and pregnancy diagnosis in farm and other animals.

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