There are many systems available for the breeder to calculate the most optimum time for inseminations, but most accurate by far is Blood Serum Progesterone levels attained by RIA (Radio Immuno Assay) or EIA (Enzyme Immuno Assay). CLONEusa highly recommends the RIA method as you do not have to test the bitch every day or even every other day which is very costly and time consuming. CLONE uses only this method because it is the most accurate and you can pinpoint the time of ovulation with only a couple of progesterone tests.

There is NO set progesterone value for breeding a bitch such as 8, 12, 18, 25 ng/ml etc. Some bitches must be bred at 6 ng/ml and some bitches as high as 50 ng/ml. Each bitch has to be treated as an individual and should be bred between 48 and 72 hours after you calculate the day of ovulation. The reason for this is that when a bitch ovulates her eggs are immature and can not be penetrated by sperm until they are mature, which in the dog takes between 40 - 60 hours.

CLONE was the first company to use progesterone testing in bitches, with over 25 years research and experience, show that the majority of bitches ovulate between 3 - 5 ng/ml and a few ovulate between 6 - 10 ng/ml. However,  there is only on average a 24 hour period of difference between these two groups of 3 -5 and 6 - 8ng/ml. (the earliest and the latest ovulating bitches) REMEMBER,  THIS IS NOT WRITTEN IN STONE........EVERY DOG IS DIFFERENT!!!   By testing the bitch to pinpoint her most fertile period and using a specialized technique and shipping system, the conception rate has paradoxically improved over the years.


                   Artificial Insemination.
FRESH CHILLED SEMEN for Artificial Insemination:

When using Chilled or Frozen Semen the first thing to know is that it should be inseminated directly into the uterus for the best success. This calls for special proceedures and training:

Since chilled and fresh semen live far longer than thawed frozen semen in the uterus the best time to breed with fresh chilled semen is 36 - 48 hours after the blood serum progesterone level shows ovulation as the semen can be inseminated in advance of the maturation of the ovum/eggs.

If you are going to breed twice, breed first time 24 - 36 hours after ovulation and
then again the next day. There is no reason to skip a day as you did with a
natural breeding, because you have pinpointed the time of the most fertile period.
If there are complications with shipping and arrangements due to holidays you
could also breed at 48 hours after ovulation and then again 24 hours later.

We HIGHLY recommend that unless you are VERY experienced and educated in artificial insemination that the procedure be done by your veterinarian or a veterinarian that is experienced in the proceedure and tests required prior to the proceedure.  Multiple options are available for Chilled Semen AI (Intrauterine Surgical insemination offers the highest success percentage with progesterone testing, however several vets and breeders are successful with vaginal insemination given the proper tests are performed to ensure the timing is right for insemination).

Trans-cervical inseminations which is by far the best method, but a difficult
method to master. Fiber Optical Insemination is also used in
certain places but demands an expensive investment in

Surgical Insemination Intrauterine this the second best method where a small
incision is made in the abd wall and the sperm is inserted
directly into the uterus. Can used where there is no skill of
Trans-cervical Insemination and no Fiber Optics available and
can be stressful for the bitch as it requires the female to be put
under anesthesia or deep sedation depending on the vet.
Vaginal Artificial Insemination performed when a special Insemination like
Trans-cervical or Surgical Insemination right into the uterus is
not necessary. * The sucess rate is not as high as the surgical
unless you are accurate on the ovulation time and window for
the eggs to mature. This can be acheived by doing the
recommended Progestrone serum and  LH Surge Tests along
with Vag swabs and checks thru your vet.
The insemination with an approved Chilled Semen Technique
can done this way and it is a simple performance where you
draw the semen or the extended chilled semen into the AI
straw, inserted in the opening of the cervix and the semen is
deposited there. With a gloved hand the "Fanning" process is
mimiced while the rear of the female is kept elevated for 10
minutes to let the semen run down.
FROZEN SEMEN for Artificial Insemination:

FROZEN Semen should ALWAYS inseminated directly into the uterus. With Frozen or chilled, timing is everything. This calls for special proceedures and training on both the Stud owner and their vet as well as the Inseminating Vet.  Frozen Semen must be kept at -190 degrees even during shipping and must be shipped in a specialized container (can be stored for extended periods of time in liquid nitrogen at a temp of -300 degrees at approved frozen sperm banks for a monthly rate).

With frozen semen, the semen is deposited directly into the uterus and you breed a little bit later - that is approx. 60 - 72 hours after the blood serum progesterone level shows ovulation. This is the time when most of the eggs are mature and ready for fertilization. Thus the sperm can immediately penetrate the egg as the sperm are not depended upon to live an extended period of time. Since the cervix dilates at the optimum time of insemination, trans-cervical inseminations with the Norwegian Catheter is by far the best insemination method as the cervix can be palpated to see if it is dilated, thus confirming if the insemination is being performed at the optimum time. This palpation can not been done with any other method not even endoscopic inseminations.
I bet you are wondering "CAN I GET ALL OF THIS DONE IN TIME TO BREED?......YES, you will have enough time to get it all in if you and your vet are prepared. The ovulation for each female is individual and can vary several days, which is why the vag swabs along with the progesterone testing  are so important.  Progestrone testing is the one way breeders can know when a female dog is ovulating, thus determining the ideal time to breed.

PROGESTERONE TESTING (highly recommended for fresh, chilled & frozen):

Progesterone is a reproductive hormone that begins to increase in the bloodstream just prior to ovulation. For most bitches a progestrone reading of 2.0 nanograms indicates a surge in the Luteinizing Hormone (LH) from the pituitary gland, which triggers the release of the eggs from the follicles. When Progesterone levels reach 5.0 to 10.0 a bitch is then ovulating. 


                   * Progesterone levels from testing methods may very which is why a "quanative"
                     Progesterone level is recommended.The key is to get a baseline reading and see 
                     an increase from the level to know the female is ovulating.

                   * Within the first 5 days of Proestrus, take a progesterone reading to determine the
                      baseline level (proestrus is commonly refered to as the last days or end of the  
                      bleeding heat portion)

                   * At 50% cornification, or around the 4th or 5th day of Proestrus, start doing the 
                      progesterone testing every other day.

                   * Around the time of LH surge is occuring, the progesterone level will be increasing
                     to (each female may vary this is why the vag swabs are helpful) above 2.5ng/ml
                     which usually means the ovulation has occured, and breeding my commence
                     within 48-72 hrs (depending on the type of breeding).

                  * When the progesterone level has risen to the level of 10ng/ml the female is during 
                    her peak of fertility. PEAK IS CONSIDERED TO BE BETWEEN 5-20ng/ml on average.

                  * Progesterone will continue to remain elevated for 2-3 months in both pregnant and 
                    non-pregnant dogs.

Many Progesterone Kits Chart progesterone levels thru color changes and these
types are used by many vets.  IF YOU ARE PAYING FOR A STUD SERVICE......IT IS
rather than being told to start breeding in a certian number of days and breed 2-3
times to allow for error. Color kits will give levels in C1-C5 each group repersenting a
range of numbers. If the Quantitive Level  (exact measurement in numbers) is not
available at your vets check on the cost and expected turn around time to draw and
                 send it to another lab or location  for the numeric count. Otherwise check with other
                 vets in your area to see if they offer a numeric reading, most fertility specialists have
                 this in-house as well as the larger vet clinics.

                 In cases where it takes longer than a couple hours to obtain a quanitative
                 progesterone level (such as needing to send to another lab after collection requiring                     a 24hr period or longer to obtain results), "breeders must use combination of the
                 laboratory test  and  behavorial changes to help pinpoint the time". To accuratley
                 identify the time of ovulation, a series of progesterone tests are conducted to map
                 the rise of progesterone levels. Multiple testing becomes even more critical when 
                 chilled semen is being shipped to another state or country and when using frozen
                 FOR FERTILIZATION.

               OvuCHECK is the only approved color charted Progesterone Testing Kit        that is approved by the Synbiotics Corporation Canine Reproduction                                                                                   Specialists and is developed and manufactured by Synbiotics
            Corporation and is available for purchase by your vet.

                       *NEW VET ACCOUNTS can be established w/ Order.
  Sameday and next day shipping available via Fed-Ex


After ovulation, the released eggs continue to go through maturing process which ends 2-3 days later. ONCE THE MATURATION PROCESS IS COMPLETED......EGGS ARE READY FOR FERTILIZATION!  As a result, the prime time to breed is 48 to 72 hours after ovulation!

Once Ovulation Occurs it takes between 48-72 hours for the eggs to become mature enough to fertilize and a male's sperm can survive safely up to 48 hours inside the female which is why it is important to be percise on your insemination date.

SYNBIOTICS CORP. PROTOCOL FOR Luteinizing Hormone (LH Surge) Testing Protocol

* Within the first 5 days of Proestrus, take a progesterone reading to determine a
              baseline level.

At 50% cornification, around the 4th or 5th day of the Proestrus (ed of bleeding heat),
              start doing the LH testing daily (if available) and Progesterone testing every other day.

A Positive Status-LH test result indicates the LH level in the sample is > 1ng/ml. 
              During this peak, progesterone levels are about 1.5 to 2.5 ng/ml. Around this time the
              progesterone will be increasing.

The day of the LH surge is counted as Day ).  The surge will last approximatley 24
               hours and triggers the ovulation.  OVULATION OCCURS 2 days  AFTER THE LH SURGE
   The Fertile Period is 4-7 days post LH SURGE and lasts 48-72 hours.

Do another Progesterone test 2-4 days post LH surge to e sure that the progesterone is indeed                 increasing thus confirming that the ovulation has occured.

Witness LH was designed to pinpoint the optimum time for canine
  breeding. Since inception, LH testing has proven variety of uses in
                                                                            multiple species.
                                                              During the canine estrus cycle, the LH surge may occur from days
                                                             3 to 28 (average days 3-12) after obserable signs of heat and
                                                             lasts approximately 24 hours.  Ovulation occurs 2 days after the
                                                             LH surge.  The ova/eggs then require and additional 2-3 days to
                                                             mature into a stage that will support fertilization.
                                                            The fertile period of the bitch falls between days 4 and 7 after the
                                                             LH surge with the most fertile days being 5 and 6 (the day of the
                                                             LH sure is counted as Day 0).  Determining the LH surge is
                                                            perhaps the most critical factor in reproductive efficiency.

This is a great fertility chart that was given to me by Dr. Mallard at Strafford Vet Fertility Clinic whom is known for being the "Reproduction Specialist" of the southwest. He and his staff are who sold us on the SYNBIOTICS Corporation Products and & Techniques.  We have had high success with our own females using Synbiotic products......they have assisted in the past 2 litters being large litters of 6 and 7 puppies  using  "Vaginal Artificial Insemination" (only recommended if your doing the insemination within 6 hrs or less of collection). 

This is chart used in assistance with the progesterone test results and the physical vaginal exam findings. It shows your female's heat cycle from start to finish and assists in determining the best times to start your progesterone testing. Further more, it assists you in coordinating your efforts in obtaining your semen and knowing what day(s) are the best to breed your female according to the chemical results (progesterone levels) and the physical findings of a vaginal exam and swab that your vet may perform (watching cornification of the cells). A series of PROGESTRONE tests are the most reliable and effective way of artificial insemination and can save you money on your breeding fee. If you know within a few hours of when your females eggs are going to drop you only need to breed her once surgically and we recommend twice (2) if doing Vaginal AI procedure. This can be done by separating the initial dose of sperm into 2 doses and giving one day apart or by purchasing an extra dose at half price (which will guarantee more sperm cells making the journey to the eggs and a greater chance of all eggs released to be fertilized).

If you are not surgically implanting, we recommend that you have your vet split the semen that we send to you in the vial so that you will have 2 separate breeding's to be used a day apart for maximum breeding results (or) purchase an additional dose of semen at half price to be sent and used a day later (this process ensures that the sperm are still strong and healthy enough to make the journey to the eggs--using the additional sperm works, however the sperms lifespan is aprox 36 hours and as you get closer to the expiration time your sperm may not be as active or possibly not active enough to make the's a chance you take by splitting the initial dose). Talk to your vet about what is the best option for you and their thoughts on what would be the best choice for your situation.
  OvuCHECK® Premate  
    -An Essential Canine Reproduction Tool
    -Determine Ovulation and Predict Whelping
    -Rapid, In-House Results in 45 Minutes
    -Maximize Litter Size through
    -Improved Ovulation Timing
     -Improve Procedure Scheduling
          WITNESS® LH
Luteinizing Hormone Diagnostic Test
-Determine Ovulation
-Improve Procedure Scheduling
-Increase Conception Rates
-Predict Whelping Dates
-Assess Ovarian Status

Eau d’ Estrus Odor attractant / Synthetic Pheromone #99-4960 (by Synbiotics Corp)- is used when a female in heat is not availale. It simulates the odor a female puts off which in turn excites the male usually producing a better semen specimen. In the event we do not have a female inheat available at time of collection........this is the next best thing. We apply this to the tail area of one of our females to simulate the actual heat and encourage the male to the next level of excitement.

                 "FRESH EXPRESS" EXTENDER #99-4350 (by Synbiotics Corporation) is used in
             conjunction with the chilling process as is a "Sperm Separating Serum (SSS) also by
             Synbiotics Corp. Semen Extenders provide an energy source and buffers that enhance the
             survival of your chilled sperm cells. FRESH EXPRESS Extender by Synbiotics Corporation 
                  is used by canine fertility specialists world wide and comes highly recommended and is only
                  vet obtained. We provide add a  high concentration ratio of this extender to ensure that the
             sperm remains healthy during transit to the awaiting female. Our dilution is 1-part Sperm
                  Rich Fraction (SRF) to 2-4 parts Extender for longer lasting specimen.

            SEMEN SEPARATING SOLUTION (SSS) #99-4950 (by Synbiotics Corp.) -The highest 
            level of success is achieved when the SRF (Sperm Rich Fraction) only is used! When using                     semen for artificial insemination with chilled or frozen techniques it is VERY IMPORTANT to
            remove the pre/post ejaculate (aka) "Seminal Plasma" and "Prostatic Fluid".

                   The inclusion of seminal plasma will, in most cases, decrease the survival time
                   and fertility of the sperm.

Typically dogs produce 0.5-2.0ml of SRF (sperm rich fraction) depending on their size.  Anything more is  the Pre/Post-ejaculate (aka) Seminal Plasma and Prostatic Fluid. Semen that is extracted and NOT IMMEDIATELY USED should have the Pre/Post Ejaculate portion removed to insure maximum sperm survival.

For this technical procedure the Semen Separating Solution (SSS) by Synbiotics Corp. It is used to separate the sperm plug from the excess fluids. This is done in 3 steps:

1) During the sterile collection process the pre/post ejaculate aka seminal plasma and Prostatic 
   Fluid is discarded keeping only the sperm rich portion (SRP) of the ejaculation. This is
   checked under microscope for "MOTILITY" and "SPERM CONCENTRATION".

2) This SRP of the Semen is slowly layered on top of the Semen Separating Solution  to form
   two distinct layers and is then placed in a clinical centrifuge at a 2000rpm or 100-150G for five
   minutes (faster spinning in a centrifuge causes damage to the sperm cells).  This process
   allows for complete separation of the plasma from the sperm without causing damage to the 
   sperm cells.

3) The Seminal Plasma and SSS is then gently removed using a pipette (as much as possible  
   without disturbing the sperm pellet) leaving only a soft pellet of sperm cells.

ONCE THE SPERM PLUG HAS BEEN SEPERATED it is mixed with the "FRESH EXPRESS Extender" and placed in the inner compartment of the shipping box and allowed to chill at
5 degrees Celsius for NO MORE THAN 1 hr prior to shipping (this initiates the chilling process and ensures the sperm do not warm and burn their energy).


1- "FRESH EXPRESS COLLECTION / INSEMINATION RECORD (provided by the Synbiotics Corp for each
  extender and separation serum used) will be included in each shipment. This Record is to be
completed by both the collecting veterinarian and the inseminating veterinarian a  
copy is to be returned to us (the stud owner) as verification that the sperm arrived in a usable form and
was evaluated and inseminated by a licensed veterinarian. A copy of this completed form will also be
placed onfile at our vets office.

1- AKC FORM for AI using Fresh Extended Semen will be provided with each Stud Service.

1- Stud Service Agreement Contract (a copy to be returned to breeder within 7 days) this can be sent
   back with shipping container.

We are extremely fortunate to have a Bulldog Specialist and a Canine Reproductive Specialist as primary caregivers to our bulldogs and their needs. It defiantly makes a difference in the quality of the bulldogs and services that we offer.

We are a firm believer in the saying "healthy sperm makes better litters" (of course this also depends on how accurate your timing is on the female's end). WHEN PROVIDING FRESH, CHILLED, EXTENDED or FROZEN SEMEN SERVICES or if the ovulating female is not at our location we take special care of the collection and processing of semen.

We use only the highest rated Canine Reproduction Fertility Extender and Separator and is shipped in specialized Semen Shipping Containers. is sent in a specialized and lab approved package that has 2 layers of Styrofoam, inside you will find 2 chill packs that is designed to keep the vial at a consistent chilled temperature and an inner compartment that protects the semen vial from laying on the chill packs (which causes shock and freezing) as well as keeps it from getting broken during delivery. DUE TO THE INDIVIDUAL COST OF THESE SHIPPING CONTAINERS, THEY MUST BE RETURNED ($85 each) this allows us to keep your overall cost down and ensures that your semen will arrive to you in a temperature controlled environment.

There are only a few degrees difference in chilled and frozen temperatures, however there is a BIG difference between in the preparation process for sperm survival in chilled vs. frozen shipping. This could be the difference between successful and unsuccessful breeding. Is Semen that is prepared for Chilling is allowed to become to cold or frozen.....the sperm cells are shocked......and then die off. Thus making it overly important that approved shipping containers and materials are used.

Frozen Semen must be prepared and packaged entirely differently to ensure survival. Semen is frozen using Cryo- Extenders and Liquid Nitrogen in commercially available tanks and shipped in "dry dewars" which are small tanks. The temp for Frozen Semen MUST be maintained at -196 degrees Celsius until ready for use.
This is a unique process for collecting semen from a stud dog and express shipping the semen for the insemination of a bitch has been widely used in the United States, Europe and other countries for over 20 years by Reproduction Specialists, Fertility Clinics and Veterinarians. This process makes it easy for breeders to provide the purchasing party with the highest quality semen thus increasing a higher success rate with artificial insemination proceedures.

The semen is collected, preserved with the "FRESH EXPRESS " special solution developed by Synbiotics Corp., packed in a "FRESH EXPRESS" Specialized Shipping box  made to regulate the tempurature and protect the vial during transport.  It is then shipped by the fastest means possible. The semen will be ready to be activated and inseminated in the bitch with an approved AI proceedure (performed by your vet) on it's arrival.

    Vet processed with Speciality formula used by Canine Fertility and Reproduction Specialists across the country

Synbiotics Corporation offers a wide selection of canine reproductive services to breeders and veterinarians across the globe. Using advanced technology developed through years of research by leading scientists and veterinary specialists, Synbiotics provides the best chance of conception available today. This system includes canine-specific freezing processes, storage and shipping methods designed to protect your dog's valuable semen.

Synbiotics currently trains, manages and supports two networks of veterinary service providers. Dog owners seeking knowledgeable medical expertise in canine reproduction are referred to the participating members in one of these two networks:

The Veterinary Referral Network for Canine Reproduction consists of over 1000 veterinarians with a special interest in canine reproduction and artificial insemination technology.

The Canine Freezing Center Network consists of veterinary facilities with advanced training in the art and science of freezing, thawing, and inseminating canine semen. Use of frozen semen allows dog breeders to preserve valuable genetic lines and to conduct long distance breeding without shipping the dogs themselves.

Through the success and support of this technology, Synbiotics Corporation maintains the largest canine semen storage bank in the world.    
Melanie Miller - (417) 425-7625   Certified Breeder of AKC & FCI  Registered English Bulldogs

This page was last updated: August 3, 2011