Small animal vaginal cytology and breeding management Flashcards
Vaginal cytology can be used to visualize the response of stratified squamous epithelium of the vagina to….
Estrogen
Normal cell layer amount in vaginal cells when not under the influence of estrogen
2-4 cell layers thick
Number of cell layer in vaginal cells when under the influence of estrogen
> 20-40 cells thick
Acts as protection mechanism for the vagina against trauma (breeding)
Vaginal cytology technique in the dog
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)
Vaginal cytology samples should be observed at _____x magnification
10
Vaginal Cytology during proestrus
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
Vaginal cytology during estrus
Estrogen levels decreasing and progesterone increasing
> 90% Superficial epithelial cells
No WBC’s
Few RBC’s
Little or no debris in background of slide
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?
Estrus
Vaginal cytology during transition to diestrus
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
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?
Transitional, early diestrus
Vaginal cytology in diestrus
Influx of neutrophils and parabasalar cells
If more than occasional neutrophils are seen in a dog that is not in proestrus or diestrus, what is top ddx?
Pyometra
What is vaginal cytology useful for clinically?
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)
Vaginal cytology (does/does not) tell you when to breed the animal
Does NOT
Pre-Breeding Concerns in SA
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
Routine Breeding Management in SA
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
Dogs should be bred __-__ days until no longer exhibiting standing estrus
2-3
In the canine cycle the first rise of ____________ is correlated with LH peak
Progesterone
2 hormones necessary for the standing behavior in bitches
Progesterone rising and estrogen falling
In bitches ______ and ______ both peak at the onset of estrus
LH and FSH
Hormone profiles in SA patients can be used to track…
Prelutinization of follicles
Based on the amount of progesterone being released can help determine most fertile days
Modern standard for breeding management in SA
Progesterone
Progesterone levels in SA patients may be measure by…
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
Progesterone levels in SA should be sampled…
Every other day (M,W,F)
Progesterone level ranges and their significance
<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)
A dog has its progesterone level tested and come back at 2ng/mL. What stage of the cycle is she likely in?
LH peak (ovulation typically 24-48h after peak)
Time Frame for using
Progesterone when breeding
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
Peak fertility in dogs is typically ___-__ days from ovulation
3-4
Fertilization in SA occurs __-__ days from LH peak
3-6
Ovulation begins ___-___ hours after LH peak
24-48 (or beginning of first rise of progesterone ~2ng/mL)
In SA, ovulation takes __-___ hours to occur
24-48
LH Assays are helpful in conjunction with….
Progesterone levels
More definitive than progesterone
But requires daily samples and is highly liable
LH assays are more definitive than progesterone levels but…
Samples must be collected daily and it is highly liable
More useful for determining if ovarian remnants present)
In a dog that is spayed the LH assay should come back as being…
High since there is no negative feedback from the gonads
Progesterone testing starts on day ___-____ of the cycle
5-7
Cytology and vaginoscopy can be used as needed
Breeding based on scenario in SA
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
AI methods in SA
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
With good quality semen and a fertile bitch a ______ AI is sufficient
Vaginal
Pregnancy and whelping rates equivalent for Transcervical (TCI) and…
Surgical (SAI)
Canine Brucellosis transmission
Sex or the environment
Canine Brucellosis causes abortion due to…
Placentitis
In males Canine Brucellosis may lead to…
Orchitis/ epididymitis
Any dog being bred should be tested for…
Canine Brucellosis
Carriers often asymptomatic
Diagnosis of Canine Brucellosis
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
Interpretation of Canine Brucellosis testing
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
Feline Breeding Management
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
For 100% ovulation rate in queens they should be bred…
at least 4 times
