Normal Cycle
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Canine Vaginal Cytology
Technique
- Moisten a cotton swab with 1 to 2 drops sterile saline. Open the vulvar lips, pull the vulva doraslly, insert the swab dorsally and posterior, then up and over the pelvic brim and into anterior vagina. If you do not pass the swab far enough, you will get vestibular cells and result in false cornification. If you pass the swab too ventral, you may enter the bladder and get a falsely non-cornified smear.
- Roll the swab firmly onto a slide.
- Stain the slide using DifQuik stain, 10 dips in A, 15 dips in B, and 20 dips in C. You may also use new methylene blue stain.
- Read the slide under low power first to establish the trend of cellularity and cell types. Move to a higher power to establish the cell types. View several fields to get an overall visual idea of the percentage of cornified cells.
Natural Breeding
![[IMAGE]](http://members.aol.com/tailwynde/ties1.jpg)
the male has turned and 'stepped over' so the dogs are rear to rear.
FRESH CHILLED SEMEN
Fresh chilled semen uses energy as it is cooled to 40'F (4'C) and eventually re-warmed to body temperature. The life in-utero of spermatozoa having experienced the chilling and subsequent warming process is 24 to 72 hours, necessitating a more precise manner of ovulation timing and breeding. When a veterinarian receives a fresh chilled sample, the package should immediately be opened. Attention should be paid to the "impression of coolness". The ice packs should be at least cold, if not still frozen. The package containing the semen should be removed from the packaging material. The tube should contain the extended semen in a liquid state. Unfortunately, occasional mishandling by the shipping company or by the shipper placing the semen package in a non-pressurized compartment of the airplane will cause the sample to arrive frozen. The freezing kills the sperm cell and renders the sample useless.
One drop of the sample should be placed on a warmed microscope slide. The rest of the sample should be refrigerated. Allowing the chilled sample to warm to room temperature only allows the sperm cells to speed up, using precious energy and shortening their life span.
In many cases, the semen will appear to be non-motile on the slide. However, as the semen drop warms gradually, side to side motility becomes noted. The continued warming eventually shows the cells to have achieved a normal forward progression. Do not warm the sample to fast. Micro-scope lights will over heat the semen and kill the sperm. If no motility is noted after fifteen minutes, the sample is most likely non-viable.
It is recommended that the refrigerated fresh chilled sample NOT be Warmed to room temperature or body temperature before insemination. Having the sample in the uterus as it warms makes maximum use of the conserved energy. All fresh chilled semen samples are handled in a similar manner.
PROCEDURE
The estrous cycle for the bitch should be monitored using serum progesterone assaying or some other reliable manner to ascertain that ovulation has occurred. Semen motility is evaluated as previously described.
The bitch is positioned with her rear elevated either manually or on a breeding ramp. Care should be taken to avoid pressure on the bitch's abdomen. The semen is placed into a large syringe from the collection tube. The syringe is then placed on an insemination rod of proper length to reach the cervical os. It is important that the semen be deposited at the entrance to the cervix so that the semen can be drawn into the uterus. With gloved hands the veterinarian gently inserts the insemination rod through the lips of the vulva at an upward 45-degree angle. The rod is gently passed over the pubis and along the dorsal median fold until it is parallel with the lumbar spine and localized in the area of the cervix. If resistance is encountered the rod should be gently twisted or withdrawn a short distance then re-advanced.
When the insemination rod is properly positioned the semen should be gently inseminated. The syringe is then removed from the rod. It is not necessary to push large amounts of air into the rod nor is it normal to get semen backflow if the rod is properly positioned and the bitch is in the estrus stage of estrous. Excess air "bubbling" through the semen is detrimental to the fragile plasma membrane of the head of the spermatozoa.
The bitch is then "feathered" digitally for one minute. The rear of the bitch is maintained in an elevated position for fifteen minutes to allow gravitational feeding of semen to the anterior vagina. The bitch owner is instructed to confine the bitch or restrict her activity, which includes elimination for two hours post-insemination.
ARTIFICIAL INSEMINATION IN A DOG
A Surgical Approach To Breeding
Artificial insemination in the dog: Modern dog and cat breeding practices often include the practice of Artificial Insemination. Presented on this page is an example of surgical insemination directly into the uterus of a dog. Another technique that can be used that does not involve surgery is the trans cervical insemination. This new procedure allows semen to be deposited directly into the cervix without anesthesia. Special training is needed with the trans cervical technique in order to be certain that the semen is deposited within the uterus; this is done using an endoscope. Some of the obvious reasons for utilizing AI (Artificial Insemination) are the following:
| easy access to sperm from selected donors |
streamlining time constraints | eliminating housing limitations | financial efficiency |
| eliminates distance limitations | greater assurance of a proper insemination |
overcoming uncooperative partners |
propagating traits from |
You and your veterinarian will want to discuss the pros and cons of AI well before the predicted time the female will come into heat. Proper Prior Planning Prevents Poor Performance! Do lots of homework so that you
know what is "out there", where you can get it, how much it costs to acquire the frozen sperm, and how your veterinarian will inseminate.
One method of insemination is to infuse the semen and associated nutrient fluids directly into the deep vaginal area of the bitch. The bitch then will have her rear quarters raised for a few minutes in an attempt to have gravity assist with the dispersal of the semen as deep into the vaginal area and as close to the cervical opening as possible. Then the bitch needs to remain confined for 6 to 8 hours after the insemination. The individual sperm cells need to pass through the cervix from the vagina and into the body of the uterus. From there, they disperse further up the two "Horns" of the uterus. The uterine horns are long hollow tubes, lined by the richly vascular endometrium, and extend from the body of the uterus upward almost to each kidney. At the end of these two tubes the ovaries are situated such that when they produce eggs, the eggs travel through a tiny opening into the far end of each uterine horn. (Just another wonder of nature!) If the eggs encounter sperm cells in their travels DOWN the uterine horns, there is a chance that the egg will be fertilized by some lucky sperm cell going UP the uterine canal and then another whole series of miracles take place that just might end up as a puppy!
Whether your veterinarian chooses to do the vaginal insemination or chooses to surgically implant the sperm directly into the body of the uterus is a matter for discussion well prior to the time a decision MUST be made. The following photos will take you through the surgical procedure. From start to finish, the surgical Artificial Insemination procedure takes only a few minutes. Light general anesthesia is required. Absolute sterile procedure is an important factor. The insemination fluid must be kept warm and implanted within the uterus as soon as possible after collection from a male who is present, or soon after shipped, frozen semen is thawed.
Incision is made in the midline abdomen.
The uterus is located.
The body of the uterus is isolated and the horns are inspected.
A tiny stab incision is made in the body of the uterus.
The semen sample is drawn into a sterile syringe.
The end of the catheter is inserted inside the uterus.
The semen sample is gently infused toward each uterine horn.
The incision in the uterus is sutured closed. Usually only one suture is required.
The abdominal wall is closed.
The skin is closed with subcutaneous sutures. Sometimes there is almost no bleeding at all.
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Now the real fun starts! Sure, tell THAT to Mama.
Pyometra In The Dog
Normal Pyometra
Here's a real nasty problem . . . pyometra. The term means pus in the uterus. Any time a veterinarian is presented with a dog or cat suffering from pyometra the condition is considered serious and immediate surgery is nearly always indicated. This pus formation in the uterus results from infection, hormone imbalance or mucous buildup inside the uterus. Most dogs and cats suffering from pyometra are presented because of loss of energy, increased thirst and poor appetite. Plus a good tip-off would be a foul smelling, purulent (means pus) vaginal discharge. Most of the cases of pyometra I've seen in dogs occurred about six weeks after the bitch's last heat cycle. They may not look it on the outside, but on the inside these dogs are really sick! If that swollen, enlarged uterus happens to rupture internally, the dog will rapidly go into endotoxic shock and whatever the veterinarian does may not be enough to save the dog
Normally, even in a large dog, the uterine horns aren't much thicker than a pencil. When pyometra is present the uterus looks and feels more like a stuffed venison sausage. I've removed eight-pound uteruses that should have weighed no more than eight ounces!
For whatever reason, if your female dog or cat hasn't been spayed be alert for pyometra. The 
condition is more probable in females eight years or older and who experience infrequent or irregular heat cycles or episodes of false pregnancy. Poor appetite, increased thirst, poor stamina and vaginal discharge are cardinal signs. And some patients' white blood cell count can go from a normal of 9,000 all the way up to 75,000. X-rays often reveal two large sausage-like structures in the abdomen. It's time for surgery! These patients should almost always be operated on right now, not after work, not in the morning, not after a few days of antibiotics "to build her up."
The surgery is not a minor procedure. A patient with a uterus swollen with a foul and putrid soup, is simply carrying a bucket of poison that would eventually kill the cat or dog. Many of these patients require I.V. fluid therapy, antibiotics and nutritional support post-operatively. These pyometra patients, once recovered, act like puppies once their near death experience is over!
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| The skin in the midline of the abdomen is incised. | The abdomen is entered and the uterus identified | The uterus is very gently pulled through the incision |
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| If there is any leakage of infected contents, life threatening peritonitis may result | The ovary, ligament and blood vessels are carefully identified | The ovary is inspected to be certain that the entire ovary is excised |
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After separating the two uterine horns the body of the uterus is visualized |
The uterus is removed from the dog after careful ligation of the uterine tissues assures a clean separation |
The abdomen is carefully sutured closed layer by layer |
Radiographic
The fetal skeleton becomes opaque 43-46 days after the LH peak making radiographic diagnosis of pregnancy possible 45-48 d after LH peak.
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Bones become visible at different times (see Rendano, Current Veterinary Therapy VIII, Kirk, ed.); for example, teeth are visible approximately 4 d before birth.
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After 50 days you can count the number, estimate size, and estimate the position of fetuses.
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At LSU we routinely do a radiographic 'puppy count' to determine how many pups there are.
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This makes the attendant at whelping much more aware if whelping is done, or if assistance is needed.
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No negative effects have been seen in fetuses that were radiographed during the latter stages of gestation.
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Fetal Death Signs of fetal death can be visible to the trained eye.
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An alteration of fetal skull bone alignment with overriding or extreme deformity is a sign of fetal death.
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Intra- or peri- fetal gas accumulations indicates fetal death.
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Abnormal fetal posture such as the "ball sign" which is the increased flexion or straightening of hind limbs.
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Can you see the pregnancy in this radiograph? No one can. The bitch is pregnant, however the uterine enlargement which is visible cannot be definitively diagnosed as a pregnancy. It may be a pyometra.
On the original radiograph (the one you will see in lab), the fetal skeletons are actually visible. A radiologist could probably see them in this picture. Why did I waste your time putting this poor image in the notes?.......I worked too hard on it not to.
Ultra-sound
Ultrasound Sector Scan of Canine Pregnancy at 25 days.
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(Yeager et. al. AJVR 53, 1992)
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Gestational sacs are visible as early as 18-20 days past the LH peak.
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There is usually little doubt when the black, hyperechoic gestation sac is observed, that the bitch is pregnant.
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Ultrasound is more accurate than palpation at all stages than palpation or radiography.
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The presence of fetal heartbeat and fetal movement can accurately establish fetal viability.
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The heartbeat is normally first seen more than 23- 25d after the LH peak. Fetal movement is seen after day 34-36 past LH peak 28.
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Ultrasound is especially useful between days d 33-45, because it is too late to palpate and too early for radiographs.
Counting the number of fetuses difficult, because you can never be sure if you are seeing another fetus when you move the probe, or if it is the same one.
Small litters may make it more difficult for ultrasound to be used to diagnose pregnancy.
Ultrasound has great value in obstetrical use in the determination of fetal viability.
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The normal heart rate in the fetus is 200 beats per minute.
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An increase or decrease in the fetal heart rate indicates fetal stress.
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The absence of a heartbeat indicates fetal death.
Natural Whelping
Stage I
- Stage I of parturition is the preparatory phase for whelping. It is averages approximately 6-12 hour long but can be as long as 36 hours.
- The bitch is usually restlessness and may show nesting behavior. She is nervous, panting, anorexic, and may tremble or shiver.
- You may see a temperature drop (to 99o F) about 24 h before stage II in approximately 85% of bitches. This temperature drop is related to prostaglandin release and the abrupt decline in progesterone.
- Can be identified with uterine monitoring.
Stage II
- Stage II is the active propulsive stage when the bitch pushes the puppies out. It lasts approximately 20 minutes to 1 hour per puppy but, no more than 2 hours should elapse between each puppy born.
- Stage II usually lasts a total of 3-6 hours but, may be as long as 24 hours total.
- The pup engages the cervix and anterior vagina, which initiates the Ferguson reflex (uterine contractions). The presentation of the puppies is 60% anterior in bitch and 60% posterior in queen.
- External signs do not always match uterine contractions when uterine contractions are monitored.
Stage III
- Stage III is the expulsion of the placenta.
- You may see pup-placenta-pup-placenta or pup-pup-placenta-placenta.
- There can be 6 hours between deliveries in the bitch, and the queen can go overnight between deliveries, but use 2 hours to diagnose a dystocia.
- Differentiation between resting in Stage III and completed parturition is difficult. You may need to take radiographs or ultrasound the bitch to be sure. Best to have films taken that last week of pregnacy!
Cesarean Surgery:
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| The dog is prepared for surgery. | The skin incision is made... | and the abdominal wall is incised. |
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| The uterus is located | and drawn gently to the outside. | The pup is located |
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| and the thin uterine wall is carefully | incised to expose the pup | within the placental membranes. |
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| The pup is removed from the uterus | along with the placental tissues | and the pup is placed onto |
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| clean towels for drying and clearing | of the airway to induce breathing. | The uterus is inspected carefully. |
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| In this case the dog is being spayed. The ovaries and uterus are removed. | Post op incision... the skin is secured by underlying sutures and glue. | The pups are encouraged to nurse as soon as possible. |
