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Breeding Soundness Examination of Bulls

By Sylvia J. Bedford-Guaus, DVM, PhD, DACT

In bulls, assessment of libido is often not possible during a routine breeding soundness
examination (BSE). However, if possible, the bull should be observed serving cows to allow
assessment of his desire to breed, ease of mounting, ability to achieve erection and extend the
penis, and presence of penile deviation or other abnormalities that may prevent successful
service. Libido and serving capacity tests (scoring the number of services achieved during a
set time that a bull is in a pen with a restrained cow) have been devised but are time
consuming and difficult to standardize under field conditions. In addition, the results are
difficult to interpret in light of the variety of stocking conditions used, eg, single versus
multiple bulls or small paddocks versus large ranges.

BSE forms have been developed and should be used to ensure the systematic completion of
the examination and accurate reporting of results. The bull should be restrained in a chute for
the examination.

 Body condition should be scored, and a general physical examination conducted with
special attention paid to the feet, legs, eyes, and sheath. The inguinal rings and
internal genitalia should be palpated per rectum to detect any abnormalities, eg,
seminal vesiculitis.
 The scrotum should be palpated to evaluate the testes, epididymides, spermatic
cords, and scrotal skin. The testes should be symmetrical in size, smooth, resilient,
and freely movable within the scrotum. Cryptorchidism is considered an undesirable
heritable trait and renders a bull unsatisfactory for breeding even though his semen
quality might be acceptable. In ruminants, the epididymides run caudomedially along
the testes in a dorsoventral orientation, with the tail most ventrad. The epididymis
should have no palpable masses. The measurement of scrotal circumference (SC) is
considered a reliable predictor of paired testis weight, which in turn provides an
accurate estimate of daily sperm production and quality. The SC should be measured
at its maximal diameter, and size depends greatly on bull age and breed. As a general
rule, SC should be ≥30 cm for bulls <15 mo old, 31 cm for bulls 16–18 mo, 32 cm for
bulls 19–21 mo, 33 cm for bulls 22–23 mo, and 34 cm for bulls ≥2 yr old. However,
readers should consult the latest published breed-specific average SC measures.

Although not used routinely in practice, additional techniques to evaluate the scrotal contents
include thermography and ultrasonography. Bull testes must be 2°–6°C (35.6°–42.8°F) cooler
than core body temperature for optimal sperm production, and increases in scrotal
temperature negatively affect semen quality. Infrared thermography can be used to evaluate
scrotal temperature, which should yield a left to right symmetry with a 6° to 4°C (42.8° to
39.2°F) temperature gradient from top to bottom. Abnormal thermograms are almost always
associated with poor sperm quality and have been correlated with decreased pregnancy rates.
Conversely, evaluation of the testes by ultrasonography does not appear to improve the
predictive value of standard evaluation techniques (physical examination, palpation, SC
measurement, sperm evaluation). Therefore, it appears that the value of ultrasonography as a
tool in BSE in bulls rests on assessment and characterization of grossly detectable lesions of
the testes.

For the purpose of a routine BSE, semen is most often collected via electroejaculation; semen
can be collected with an artificial vagina (AV) in bulls trained to use one (eg, those in bull
studs). The penis usually extends during electroejaculation and should be examined at that
time for any abnormalities. If it is not extended during electroejaculation, it should be gently
exteriorized for examination by grasping the glans with a cotton gauze and, if necessary, by
putting pressure on the sigmoid flexure immediately caudal to the scrotum. Preputial wash
samples may be taken for isolation and culture of Campylobacter fetus venerealis (using
Clark’s medium) or Tritrichomonas foetus (using Diamond’s medium or a commercially
available diagnostic medium kit), in particular when investigating subfertility in bulls ≥4 yr
old.

The electroejaculator consists of a rectal probe that has a series of linear banded
electrodes connected to a variable current and voltage source. The bull is restrained in a
chute, the rectum is emptied, and the entire lubricated probe is inserted rectally with the
electrodes oriented ventrally. A hand-operated rheostat permits intermittent pulses of current
to be given as the voltage is gradually increased. The response varies considerably, but it is
common to use 2- to 4-sec pulses repeated at 5- to 7-sec intervals. After a variable number of
such stimulations, erection and protrusion of the penis may be seen, followed by a flow of
seminal fluid, or the bull may ejaculate into the sheath without protruding the penis. The
semen may be collected by any convenient method; typically a rubber AV cone inserted
within a plastic cylinder attached to an 18-in. handle, with a test tube attached to the cone, is
used. In some bulls, ejaculation is seen only after a final series of momentary pulses at 1- to
2-sec intervals. Older bulls usually require a higher voltage for ejaculation. In some large
bulls, the probe may not reach the correct areas for stimulation; having two or more probe
sizes is recommended if BSEs are to be done on a variety of sizes of bulls. The semen
collected via electroejaculation is not representative of a full ejaculate in regard to volume
and concentration, and thus the sperm production potential of the bull is based on the
evaluation of SC. Conversely, sperm quality (motility and morphology) does not differ from
that in an ejaculate collected with an AV.

If an AV is to be used, bulls are induced to mount a teaser animal (eg, restrained steer, cow,
or phantom), and the erect penis is directed into the AV by the collector as the bull mounts. In
preparing the AV, the temperature, which is a critical factor in stimulating ejaculation, is
maintained at 105°–107°F (40.5°–42°C). Temperatures up to 118°F (48°C) may assist
collections in untrained bulls. The AV should be lubricated with nonspermicidal jelly. The
typical volume of an ejaculate is 4–8 mL, and the concentration 1–1.5 billion sperm/mL. A
semen sample can be collected from some bulls by transrectal massage of the accessory sex
glands. With this technique, erection seldom occurs. After the rectum is completely emptied,
the vesicular glands are massaged with a backward motion until a few mL of fluid drop from
the sheath. The ampullae are then massaged, and an assistant collects the semen as for
electroejaculation. This method is not always successful, and the quality of the semen
emission is usually lower than the ejaculate collected by the other two methods.

Sperm concentration, ejaculate volume, and the total number of sperm in the ejaculate are
evaluated only if the sample has been collected with an AV. The semen sample should be
evaluated immediately for sperm motility. The materials that contact the sperm should be at
the same temperature as the sperm (to avoid temperature shock), clean, dry, and nontoxic.
Motility evaluation is best when the temperature of the semen is either maintained at ~37°C
(~98.6°F) during the short time before it is evaluated, or when the semen is gently warmed to
37°C before evaluation. Both gross motility (“swirl pattern”) in the semen and motility of
individual spermatozoa should be assessed; individual motility is the more accurate measure
and should be used for classification of the bull.

Gross motility is a function of sperm concentration and individual sperm motility and is
evaluated in an undiluted drop of semen placed on a slide without a coverslip and examined
at low power (~100×). The intensity of wave motion may be classified into four categories:
very good—intense swirling, rapid dark and light waves; good—slower swirling, waves not
as intense; fair—slow movement with fewer waves; or poor—very little or no swirl activity.
Individual motility is assessed in a sample diluted with warmed saline or semen extender. A
drop of diluted sperm is placed on a slide, covered with a coverslip, and examined at 200–
400×. The proportion of sperm that are moving progressively across the field of view is
estimated by finding multiple groups of ~10 sperm and estimating how many sperm are
progressive versus how many are not.

The environments in which BSE are done are variable, and there is a high chance of
temperature shock to sperm before they may be evaluated. Therefore, >30% individual
progressive motility, or fair gross motility, is considered acceptable. If the examination is
performed under optimal conditions, some associations (eg, the Canadian Bovine
Practitioners Association) recommend a minimum of 60% motility for a bull to be classified
as a satisfactory potential breeder.

Visual estimates of sperm motility are considered appropriate for bull classification in field
BSEs. However, in commercial artificial insemination centers, sperm motion evaluation is
performed using computer-assisted sperm analysis (CASA), which provides more objective
values as well as specific measures not only of percent motile sperm but also of velocities and
the track followed by each sperm. For instance, the combination of measures such as beat
cross frequency, linearity, average path velocity, straightness, and curvilinear velocity have
been correlated with bull fertility. Moreover, some of the motion measures evaluated by
CASA such as curvilinear velocity, straightness, and linearity correlate with sperm function
such as the acquisition of hyperactivated motility.
The presence of cells other than spermatozoa in the sample should be investigated
while estimating motility. RBCs, WBCs, and excess numbers of round epithelial cells and
developing forms of spermatozoa may indicate a genital tract abnormality. Careful evaluation
of the tract, especially the internal genitalia, may indicate the source of WBCs. The most
common cause is seminal vesiculitis. Round, immature germ cells and spermatozoa with
proximal cytoplasmic droplets may indicate immaturity or, alternatively, testicular
degeneration. For a better assessment of the types of cells present in semen, staining an air-
dried smear of fresh semen with Romanowski stain is recommended. This simple technique
can provide critical information, in particular when cells other than mature sperm are present
in the sample.

A sample of the semen should be fixed in a buffered formaldehyde saline solution to evaluate
sperm morphology. This is best done with the fixed, unstained sperm examined under high
power (1,000×) using a phase-contrast microscope. At least 100 sperm should be counted,
and the proportion of sperm showing different types of abnormalities noted. However, the
only number used in the evaluation of the bull is the proportion of normal sperm, which
should be >70%. Sperm plasma integrity, which is required for sperm function, can be
assessed with the use of a vital stain such as eosin. Eosin penetrates cells with a disrupted
sperm membrane, which appear pink on evaluation under light microscopy, in contrast to live
cells, which do not stain. Eosin is often combined with nigrosin, which provides a dark
background that allows better observation of live/dead sperm cells. An eosin-nigrosin–stained
smear can also be used to assess sperm morphology, although it is considered less reliable
than using phase-contrast microscopy of an unstained wet preparation. Other, more
sophisticated measures of sperm function are not used routinely in cattle BSEs, although they
may become more common in referral and artificial insemination centers. Such tests may
include combinations of fluorescent probes and flow cytometry to assess membrane integrity,
mitochondrial function, or sperm chromatin integrity, and functional assays such as ability of
sperm to undergo in vitro capacitation, zona binding, and/or homologous in vitro fertilization.

Bulls are classified as “satisfactory potential breeders” if they have no physical abnormalities
that would prevent breeding and if they meet the minimal qualifications for SC, sperm
motility, and sperm morphology. Bulls that do not meet these criteria are classified as
“unsatisfactory potential breeders” or, if the results are marginal or questionable, they are
considered “classification deferred” and a retest should be recommended.

Breeding Soundness Examination of Rams


By Sylvia J. Bedford-Guaus, DVM, PhD, DACT

In rams, the breeding soundness examination (BSE) is performed much as for bulls, using an
electroejaculator with a smaller (ram) probe. Body score is classified as 1–2 questionable
(underconditioned); 3–4 satisfactory; and 5 questionable (overconditioned). Any major
abnormalities of the external genitalia or lumps or irregularities of the testes or epididymides
render the ram unsatisfactory. Epididymal masses are commonly found to be sperm
granulomas caused by infection with Brucella ovis, which is a major cause of reproductive
loss in sheep. B ovis may also be associated with testicular atrophy. Scrotal circumference
should be ≥28 cm for rams 8–14 mo old (>36 cm is exceptional) and ≥32 cm for rams >14
mo old (>40 cm is exceptional). Scrotal circumference lower than satisfactory is
questionable. Motility of individual spermatozoa is evaluated as for bulls, with >70%
progressive motility being exceptional, >30% satisfactory, 10%–30% questionable, and 0%
unsatisfactory. The percentage of morphologically normal sperm should be >50%;
between 30% and 50% is questionable, and <30% unsatisfactory; >80% normal sperm is
exceptional. Presence of >5 WBCs per high power field is questionable (WBCs are correlated
with B ovis infection). An ELISA test for B ovis should be done on range rams and rams >9
mo old. A positive test renders the ram unsatisfactory. Suspect tests should be repeated. (Also
see Ram Management.)

Any ram with one unsatisfactory rating in any parameter is classified “unsatisfactory”; a ram
with a questionable rating in any parameter is “questionable.” For a ram to be classified
exceptional, he must have exceptional ratings in scrotal circumference, sperm motility, and
sperm morphology. All other rams are considered “satisfactory.”

BSE

Pemilihan pejantan:
1. Uji performan:berat badan, panjang badan, tinggi gumba, lingkar dada, libido,
kualitas semen dan penyakit
2. Uji progeny: keturunan

Penampungan semen
Sapi :12 bulan
Domba: 7 bulan
Metode:

1. Massage
2. Vagina buatan
3. Elektroejakulator

1. Penampungan rata-rata dilakukan setiap 2 minggu dengan bantuan pemancing betina,


sesama jantan atau pantom

2. Ternak dimandikan dan lokasi penampungan didesinfeksi terlebih dahulu

3. Sebelum penampungan, dilakukan false mounting 3-5 kali untuk meningkatkan libido

4. Siapkan vagina buatan dengan suhu mendekati suhu tubuh 45 C (35-55C), kemudian
diberikan pelicin pada vagina buatan

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