Vaginal Cytology 12

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PROCEDURE  12-1   Obtaining Vaginal Samples

PURPOSE 1 Ovary

To obtain samples from the vagina for cytological evaluation

INDICATIONS
1. To determine whether a bitch with a hemorrhagic vulvar
discharge is “in heat”
2. To assess the degree of estrogen influence in a breeding
bitch during a heat cycle
3. To determine the first day of diestrus for timing parturition
Body of
4. To differentiate between mucoid, septic, and nonseptic uterus
vulvar discharges

CONTRAINDICATIONS AND CONCERNS

Vagina
1. Vaginal cytology can determine the degree of estrogen
influence, but does not predict ovulation date. Urethral opening
2. Poor technique can result in vaginal swabs that are not
representative of true superficial vaginal cytology.
Vestibule

SPECIAL ANATOMY
Clitoris
The caudal portion of the vagina is the vestibule, which Clitoral
extends from the vulvar labia to the cingulum, a narrow- fossa

ing just anterior to the urethral papilla. The vestibule is Vulva

angled steeply dorsocranially.
Anatomy of the caudal portion of the reproductive tract in a bitch.
     

1 When the vulvar lips (labia) are parted, the clitoris can
be visualized within the ventral commissure of the vul-
var lips. When passing swabs or a scope, it is important
to begin its insertion near the dorsal vulvar commissure
Colon to avoid the sensitive clitoral fossa.
  

Vestibule

Urethral
opening

Vestibule
Cingulum

Vagina Clitroral
fossa
Clitoris Bladder Clitoris
Clitoral fossa

Anatomy of the clitoral fossa and clitoris.

  

189 
 190 CHAPTER 12  Vaginal Cytology

PROCEDURE  12-1 Obtaining Vaginal Samples—cont’d

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3. The urethral tubercle is located on the ventral wall (floor) Gently part the vulvar lips and insert the swab at the
of the anterior vestibule. dorsal commissure of the vulva.
  
EQUIPMENT 2
• Cotton-tipped swabs
• Otoscope cone
• Glass microscope slides
• Syringe with saline
  

Inserting the swab at the dorsal commissure of the vulva.

  

Equipment needed to make vaginal cytology slides from a bitch. Advance the swab dorsally and angle slightly cranially
until the swab goes over the ischial arch, then advance
slightly cranially.
TECHNIQUE   

Moisten a cotton-tipped swab with saline. 3A

  

Moistening a cotton-tipped swab with saline.

  

PROCEDURE  12-1 Obtaining Vaginal Samples—cont’d
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3B
The swab is advanced craniodorsally until the swab goes over the
ischial arch, then it is advanced slightly cranially.
Alternative technique: If the dog is large enough, pass
an otoscope cone up through the vestibule into the va-
gina and use the otoscope cone as a speculum. Pass the
cotton swab up through the speculum so that it comes
into contact with the dorsal wall of the posterior vagina.
The advantage to using a speculum is that you sample
cells only from the vagina and not from the vestibule.
Cells in the vagina are more responsive to changing
hormone levels than cells in the vestibule.
  
4A
PROCEDURE 12-1  Obtaining Vaginal Samples 191
4B
An otoscope cone can be used as a speculum for vaginal cytology
swab collection, bypassing the vestibule.
5. Gently roll the swab against the dorsal vaginal surface,
then pull it straight out.
6. Roll the swab on a glass slide, allow to air dry, and stain
with Diff-Quick or Wright–Giemsa stain.
COMPLICATIONS
None.
RESULTS
Vaginal cytology normally varies with the stage of the
estrus cycle and the degree of estrogen influence.
A.  During proestrus there are small, round para-
basal cells with large darkly stained nuclei as well
as slightly larger intermediate cells and red blood
cells.
1A
Vaginal smear from a bitch in proestrus contains red blood cells,
parabasal cells, and intermediate cells. (Courtesy Dr. Klaas Post,
University of Saskatchewan.)
 192 CHAPTER 12  Vaginal Cytology
PROCEDURE  12-1 Obtaining Vaginal Samples—cont’d
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B. During estrus the proportion of superficial mature
cornified cells increases. Superficial cells are po-
lygonal, with a small round nucleus that becomes
pyknotic over time. Eventually the cells become
anuclear.
  
1B
Vaginal cytology during estrus consists primarily of superficial
mature cornified epithelial cells. (Courtesy Dr. Klaas Post, University
of Saskatchewan.)
2. Vaginal cornification of 50% to 60% has been recom-
mended as the best time to begin sequential progesterone
testing to determine the optimal days for breeding. An
initial rise in progesterone (>2 ng/ml) is seen at the time
of the luteinizing hormone (LH) peak, which typically
precedes ovulation by 48 hours.
  
When ovulation occurs, vaginal cytology reveals almost exclu-
sively superficial mature cornified epithelial cells and the pro-
gesterone level is further increased (>4 ng/ml). The ovulated
eggs take approximately 48 hours to mature, so the optimal
time for fertilization is 2 or 3 days following ovulation.
At the onset of diestrus there is an abrupt transition of
vaginal cytology from late estrus (80% to 100% super-
ficial mature cornified cells) to diestrus (80% to 100%
parabasal and intermediate cells plus neutrophils). On-
set of cytological diestrus is usually 4 to 6 days following
ovulation, suggesting that it is too late to breed during
the current cycle. Parturition, if a bitch becomes preg-
nant, typically occurs 58 (±1) days after the first day of
cytological diestrus which corresponds to 62 to 64 days
after ovulation.
Diestrus is characterized by an abrupt transition of vaginal cytology
to predominantly parabasal and intermediate cells plus neutrophils.
(Courtesy Dr. Klaas Post, University of Saskatchewan.)
  
4. When vaginal cytology performed in a spayed female
dog that is showing signs of estrus (bloody discharge,
attractive to males) demonstrates estrogen influence
(cornified cells), this suggests that an ovarian remnant
may be present.