Artificial Insemination

• Surgical AIs with dual sires
• Unsuccessful frozen AI
• Predicting success for first time frozen AI
• Frozen AI for a maiden bitch
• Timing a bitch bred via AI
• AI as a solution to slip ties mating

Surgical AIs with dual sires

Q: What is the success rate of obtaining pups from both sires when doing a surgical AI with dual sires?

A: While I personally have not done a surgical AI with dual sires, many in the Canine Reproduction group that I run on Facebook have. Their greatest success rates come from injecting the semen of Dog A in one uterine horn Dog B semen in the other. By doing this they are giving the semen maximum opportunity to fertilize the eggs without competing with each other. 

Gayle did a somewhat successful and unplanned dual-sire breeding using frozen semen and trans-cervical inseminations. The semen from both males was very poor, with motility below 20%. The first TCI was done on Day 6 after the LH peak. Since the semen was so bad, she decided to use the second sire the next day so another TCI was done on Day 7 post LH. She got 3 pups from the first TCI and one from the second.

The usual challenge with dual-sire breedings is that one dog’s semen is better than the other. If the breeding with the weaker semen isn’t optimized in some way, the pups are most likely going to be sired by the stronger dog. Timing the breeding closely and giving the weaker semen a boost by getting it closer to the eggs at the right time, using surgical or trans-cervical inseminations can improve the success of dual-sire breedings.

Dual-sire breedings also require paternity DNA testing of both sires, dam and all pups, as well as additional registration fees.

Unsuccessful frozen AI

Q: This resulted in no pregnancy. What happened? (Maiden Bitch)

• Jan 21/21 *Blood start
• Jan 28  *0.5
• Feb 01  *3.8
• Feb 02  *6.6
• Feb 04  *TCI fresh chilled

Semen was great quality

A: I actually have more questions than suggestions! In order to help you it is important to know what unit of measure was used for the progesterone, was it nmols or ng? Then, you did not run further progesterone testing to even know that she was over 10ng which is one of the lower numbers that is able to sustain a pregnancy. There is a distinct possibility that your girl could have stalled out, and never gotten higher, so this is why I always do two things. First, I make sure that I save 0.25-0.5cc of serum in a red top tube, label and freeze so that I can run an LH test to be sure I know when she ovulated. Then, second, I run additional progesterone numbers. I run one on the day she is bred and another a day or two later just as insurance. When doing a TCI you are placing that semen right where it was supposed to go so I actually look for a higher number, 20ng+ to do the insemination on my girls. IF she does not go higher, I would be looking for possible cysts in the reproductive organs that can negatively impact hormones and the ability to become pregnant and maintain a pregnancy to term.

Semen quality, you say it was great quality, but you did not share any numbers. The quantity of sperm matters as does the forward motility, and the number of abnormal sperm. Semen quality is comprised of various components, and it is important to know what each of them is.

Predicting success for first time frozen AI

Q: I have a 6 1/2 yr bitch never whelped before. Going to try frozen. Been screen tested and in perfect health. What do you think the odds are of getting pregnant? 

A: Given your scenario, this is what I would do. First, working with my reproductive veterinarian, I would run a full bloodwork panel, including T3 and T4. You want to make sure that she is in optimum health. It sounds like you have already done your breed health testing and that she has passed with flying colors, so that is great!

Once she comes into season, I would start testing perhaps a bit earlier, just in case, starting on day 4-5. I would also be prepared to run a significant number of progesterone tests because you want to make sure you catch the LH surge and ovulation. With every blood draw for progesterone, I make sure that I save 0.25-0.5cc of serum in a red top tube, label and freeze, so that I can run an LH test to be sure I know when she ovulated. Then, I run additional progesterone numbers that others might not. I run one on the day she is bred and another a day or two later just as insurance. The reason for this is that I want to know she is rising high enough to sustain a pregnancy. Remember when you are doing a frozen breeding you have a very small window as the frozen semen does not have a long life once unthawed and inserted. It is actually a maximum of only 12-24 hours, so you need precision timing, which is why all the blood draws for progesterone and LH.

Semen, especially frozen, is a fragile thing and you only have one shot with SI, unless you are doing two TCI’s with additional frozen. Therefore, you want to work with your vet to make sure that you look at the post thaw report, and factor in that it will probably drop from what it was initially. If you have any doubts as to the motility, abnormal sperm or quantity post thaw I would request another straw to make sure you give every opportunity for conception.

Frozen AI for a maiden bitch

Q: I have a 3.5 year old maiden bitch I would like to breed at her next cycle. I am having difficulty finding a live stud that can bring to the breeding what I'd like to improve in my bitch, and still not be a heavy line breeding. I do have frozen available that would be a great match, but I have always heard that it's best to do a live cover breeding for a maiden bitch. I'd love to hear your thoughts on using frozen for a maiden bitch.  

A: I have a “love-hate” relationship with frozen semen, and I am in the middle of doing a frozen TCI/SI with my own maiden Gordon Setter girl as I respond to this. If given the preference I would personally always use frozen on a proven bitch, as that way I am sure that she is capable of getting pregnant, carrying to term, and is a good mother. However, sometimes you just need to use what is best for the breeding. When this is the case, you want to ‘cross all your T’s’ and ‘dot all of your I’s’ to do whatever is necessary to be successful. I would start with a full veterinary exam, blood work including blood chemistry, thyroid T3 & T4. This gives you a good baseline to know that you do not have anything hidden that could cause problems. I would also want to know that she and her mother have normal regular cycles, as fertility can be hereditary.

 Doing this breeding, I would work with a reproductive veterinarian who will also be able to run LH surge testing for ovulation and run regular progesterone testing as well. The reason for this is that you have a super small window for breeding, only 8-10hrs, so you have to get it right. For my Gordon Setter we ran progesterone every day, saving 0.25cc-0.5cc of serum labeled and frozen so we could run LH testing when we saw her progesterone jump. By doing this we were able to accurately know the date of ovulation/LH surge. We counted 5-6 days post LH surge and scheduled a TCI, prepared to flip to SI if there were any issues in the TCI. We did the TCI successfully on day 5 ¾ from LH surge, and ran another progesterone early in the day, with a result of 26.4ng. This let us know that the eggs were ready, and the timing was optimal. Now we cross our fingers! These tips will help you evaluate your situation so that you can make a good decision.

Timing a bitch bred via AI

Q: I have a 4 year old bitch who was bred via AI and according to the vets progesterone tests for correct timing. The semen used had a good count and motility, yet she did not conceive. Are there any tests or procedures that are used for the bitches? Is TCI the next recommended procedure?

A: You do not say if your girl has previously been bred and carried a litter. So, at this point I have more questions than answers. This is important as her age of 4 years old can be a factor if she has not yet had a successful pregnancy and carried to term. While you did progesterone testing, did you also save serum each day and freeze it in order to do LH timing too? I think we often breed too early, and so I now wait until my progesterone numbers are between 10-22ng to breed, and sometimes even higher depending on the number of days past ovulation and LH. Did you run complete bloodwork on her, including T3 and T4? Also did you ask for a guarded vaginal culture? After she was bred, did you run any further progesterone to see if she was able to maintain a high enough level to stay pregnant? These are things that can help pinpoint any possible problems. I have a girl who never ever got pregnant if there was any extender used on the AI. When I did an AI with her, it always had to be side-by-side, and I mention this simply as something to consider.

If getting her pregnant is the ultimate goal, I would make sure that I do all of the steps that I mentioned above, and most likely I would do a Surgical Implant (SI) if this was my girl. Doing an SI allows your veterinarian to inspect the uterus and the ovaries for any physical issues, including cysts. 

AI as a solution to slip ties mating

Q: Question on slipped tie mattings. I have a female that has never had issues with studs and ties...but trying a new stud with her and he slipped out every time! Had them together every other day over what is her normal 6 days of mating (days 10-15) Finally had a 2-min tie at the very end but that was after he attempted and slipped out 2-3 times an hour earlier. His knot did swell at every attempt as it was in that moment he would "slip.” What are the chances she is pregnant? There was a total of 10 slips. Follow up, can males be prone to this? As I have another female who is coming into heat who did have a litter with him back in early winter after they did finally tie back then (short ones - like 6-8 min ones)...but now I'm concerned with him that he's going to slip with her as well this time around? Thoughts? 

A: My first question is whether or not your vet did a full morphology, semen analysis on the stud dog, as well as a wellness check to make sure he could get the job done? Slip ties frequently result in pregnancy, so I am not overly worried about that. What is most important is his ability to plant the semen in her so that it has a fighting chance to fertilize the eggs. That is where the tie comes in as it stimulates the female’s muscles to pulse, helping them along their way and also plugging the exit! I would try holding the girl so that she is positioned across your knees and give your male an assist so that he can penetrate and tie with her. Optionally, you can collect him and do artificial insemination (AI).