Small animal vaginal cytology and breeding management

Question 1

Vaginal cytology can be used to visualize the response of stratified squamous epithelium of the vagina to….

Answer 1

Estrogen

Question 2

Normal cell layer amount in vaginal cells when not under the influence of estrogen

Answer 2

2-4 cell layers thick

Question 3

Number of cell layer in vaginal cells when under the influence of estrogen

Answer 3

> 20-40 cells thick
Acts as protection mechanism for the vagina against trauma (breeding)

Question 4

Vaginal cytology technique in the dog

Answer 4

Start with swab perpendicular to the ground
Put swab all the way up until resistance is felt
Then angle swab to ~45-90 degrees and advance further
Twist the swab in one direction and then pull it out maintaining the 45 degree angle

WANT TO AVOID URETHRAL SAMPLE (false positives) (do not enter initially at a 45 degree angle)

Roll sample onto slide in one direction, make ~3 lines on the slide

Stain with diff quick

Scan slide at 10x magnification (rarely need to go above)

Question 5

Vaginal cytology samples should be observed at _____x magnification

Answer 5

10

Question 6

Vaginal Cytology during proestrus

Answer 6

Estrogen is at HIGHEST

Superficial epithelial cells are cornified (like corn flakes)- large angular cells with a small or absent nucleus

Red blood cells - due to diapedesis from uterus

White blood cells - PMN’s - variable in number - decreasing in amount over
time

Debris in the background

Question 7

Vaginal cytology during estrus

Answer 7

Estrogen levels decreasing and progesterone increasing

> 90% Superficial epithelial cells
No WBC’s
Few RBC’s
Little or no debris in background of slide

Question 8

You are observing a vaginal cytology from a female dog. You observe >90% superficial epithelial cells with little debris in the background of the slide and 1-2 RBC’s. What stage of estrus is the dog in?

Answer 8

Estrus

Question 9

Vaginal cytology during transition to diestrus

Answer 9

Vaginal cytology - abrupt change to parabaslar cells, influx of neutrophils

Vulvar swelling decreases

Vulvar discharge disappears - sometimes see scant purulent discharge
early (normal)

Progesterone peaks at 3 weeks
Prolactin - luteotropic

Question 10

A dog presents to you with scant purulent discharge and minimal vulvar swelling. On vaginal cytology you observe large amounts of neutrophils that are non-toxic and parabasalar cells. What phase of the estrus cycle is this dog in?

Answer 10

Transitional, early diestrus

Question 11

Vaginal cytology in diestrus

Answer 11

Influx of neutrophils and parabasalar cells

Question 12

If more than occasional neutrophils are seen in a dog that is not in proestrus or diestrus, what is top ddx?

Answer 12

Pyometra

Question 13

What is vaginal cytology useful for clinically?

Answer 13

Influence of estrogen (superficial epithelium becomes cornified)

Is there inflammation present? (neutrophils)

Determine accurate due date
-Switch from superficial epithelial cells to parabasal cells (57 1/2 days after shift)

Question 14

Vaginal cytology (does/does not) tell you when to breed the animal

Answer 14

Does NOT

Question 15

Pre-Breeding Concerns in SA

Answer 15

Health testing?

General Health Care
 Vaccine status
 Internal and external parasite control
 Diet, activity level, BCS score

Discuss upcoming breeding
 Plan well in advance!
 Have back-up plans

Question 16

Routine Breeding Management in SA

Answer 16

Identify first day of vulvar discharge
Start counting = Day 1

Identify first day of standing estrus

Breed every 2-3 days until no longer stands

Average scenario:
Breed on Day 10, 12, and 14
Out of estrus

Question 17

Dogs should be bred __-__ days until no longer exhibiting standing estrus

Answer 17

2-3

Question 18

In the canine cycle the first rise of ____________ is correlated with LH peak

Answer 18

Progesterone

Question 19

2 hormones necessary for the standing behavior in bitches

Answer 19

Progesterone rising and estrogen falling

Question 20

In bitches ______ and ______ both peak at the onset of estrus

Answer 20

LH and FSH

Question 21

Hormone profiles in SA patients can be used to track…

Answer 21

Prelutinization of follicles
Based on the amount of progesterone being released can help determine most fertile days

Question 22

Modern standard for breeding management in SA

Answer 22

Progesterone

Question 23

Progesterone levels in SA patients may be measure by…

Answer 23

Semi-quantitative Enzyme-linked immunosorbent assay

Quantitative methods preferred:
 Radioimmunoassay – RIA (hard to find these days)
 Chemoluminescence

 Others exist, but are not consistent
 IDEXX Catalyst machine has not been independently validated and currently is found to be highly INACCURATE above 3ng/mL

Question 24

Progesterone levels in SA should be sampled…

Answer 24

Every other day (M,W,F)

Question 25

Progesterone level ranges and their significance

Answer 25

<1ng/mL- No luteal tissue present 2ng/mL- LH peak 5ng/mL- ovualtion (large range can also look for double in level within 48h period)

Question 26

A dog has its progesterone level tested and come back at 2ng/mL. What stage of the cycle is she likely in?

Answer 26

LH peak (ovulation typically 24-48h after peak)

Question 27

Time Frame for using Progesterone when breeding

Answer 27

Ovulation begins 24-48 hours after LH peak or the first rise in progesterone (~2ng/ml) Ovulation takes 24-48 hours -wave of ovulation over time for multiple follicles Two meiotic divisions must occur to be fertile -primary oocytes to secondary oocytes 24-48hours 3-6 days from LH peak – Fertilization 3-4 days from ovulation– peak fertility

Question 28

Peak fertility in dogs is typically ___-__ days from ovulation

Answer 28

3-4

Question 29

Fertilization in SA occurs __-__ days from LH peak

Answer 29

3-6

Question 30

Ovulation begins ___-___ hours after LH peak

Answer 30

24-48 (or beginning of first rise of progesterone ~2ng/mL)

Question 31

In SA, ovulation takes __-___ hours to occur

Answer 31

24-48

Question 32

LH Assays are helpful in conjunction with....

Answer 32

Progesterone levels More definitive than progesterone But requires daily samples and is highly liable

Question 33

LH assays are more definitive than progesterone levels but...

Answer 33

Samples must be collected daily and it is highly liable More useful for determining if ovarian remnants present)

Question 34

In a dog that is spayed the LH assay should come back as being...

Answer 34

High since there is no negative feedback from the gonads

Question 35

Progesterone testing starts on day ___-____ of the cycle

Answer 35

5-7 Cytology and vaginoscopy can be used as needed

Question 36

Breeding based on scenario in SA

Answer 36

Natural– EOD as long as she is standing -Shipped chilled– Error on the early and not late -semen lasts for 5-7 days Frozen– need to catch on most fertile days - d 3-4 post ovulation or 5-7 post LH -Semen will last 12-24 hours, if lucky

Question 37

AI methods in SA

Answer 37

Vaginal - Large animal AI pipette - Labial folds to just cranial to the illium ensures semen deposition in cranial vaginal vault Transcervical (TCI) - Endoscope-guided catheritization of the cervix and deposition of semen directly in the uterus Surgical (SAI) -Laparotomy with semen deposition via injection within the uterine lumen With good quality semen and a fertile bitch a vaginal AI is sufficient Pregnancy and whelping rates equivalent for TCI and SAI

Question 38

With good quality semen and a fertile bitch a ______ AI is sufficient

Answer 38

Vaginal

Question 39

Pregnancy and whelping rates equivalent for Transcervical (TCI) and...

Answer 39

Surgical (SAI)

Question 40

Canine Brucellosis transmission

Answer 40

Sex or the environment

Question 41

Canine Brucellosis causes abortion due to...

Answer 41

Placentitis

Question 42

In males Canine Brucellosis may lead to...

Answer 42

Orchitis/ epididymitis

Question 43

Any dog being bred should be tested for...

Answer 43

Canine Brucellosis Carriers often asymptomatic

Question 44

Diagnosis of Canine Brucellosis

Answer 44

Multiplex from cornel- Current gold standard PCR (best for active infection) Culture- VERY DIFFICULT but one of the gold standards Agar gel immunodiffusion test (confirmatory) IFA

Question 45

Interpretation of Canine Brucellosis testing

Answer 45

False negatives: Early in disease process - < 4-6 weeks after treatment False positives: Lack of specificity of tests Cross-reactivity with other antigens If negative on routine slide test: No other test needed If positive results on slide test: AGID to rule out false positive or PCR If positive with clinical signs: Culture of Blood/ discharge or PCR If negative with clinical signs: 3 monthly tests required to rule out false negative

Question 46

Feline Breeding Management

Answer 46

Natural breedings the norm Copulation necessary for ovulation - unless GnRH/hCG given Multiple breedings ensure complete ovulation (4 for 100% ovulatory rate) Bring queen to male’s territory – allow time for acclimation

Question 47

For 100% ovulation rate in queens they should be bred...

Answer 47

at least 4 times