Canine Progesterone Testing

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 Take your breeding program to the next level with the Canine P4 Analyzer

The P4 Analyzer is a simple device made for humans but I was able to calibrate it to work in animals, I ran test with each lab machine mentioned below, and by working side by side to one another I was able to get the formula for each device, this machine is simple to calibrate, just with a move of an up and down button, It never comes in contact with liquids so no service is ever needed.

It took me over a year but was able to make it work, I was able to sometimes get to .5 n/g difference on each machine,  I was able to calibrate to an Elisa, a MiniVidas / Antech and also  Idexx, I have a conception rate of almost 98% now and for our c-sections I´m right on time, following the reverse progesterone from day 59 until they drop below levels to 3.0 n/g or less I know the bitch is ready and the puppies lungs are ready for life.

I have been breeding English bulldogs since 1996  and I have always wanted a progesterone machine that  I could use at home and not have to be transporting my females from place to place or having to call a 2 Vet at 2:00 am or 4:00 am, I also got tired of using the Target Test or the color changing test that were not always accurate, I also bought the Bio-vet machine that takes over an hour to get results and also having to mix reagents and calibrate every time I wanted to run a test and ended up wasting 5 wells in order to run one test so that was not very cost effective and very time consuming.

With the P4 Analyzer after you have serum separated from the blood, it takes 15 minutes of incubation time in the P4 Chamber and all you do is put the strip in the machine and get the results in seconds,so this is very fast easy and affordable.

Dog Progesterone Test-Information on Dog Progesterone – Canine Progesterone testing-In-house Progesterone testing

These machines are not made in USA, I ship them via UPS and takes about 3 to 7 business days from the day I ship to clear customs, all of our test kits are available for next day air if needed, we have all stock and consumables in Miami, Fl.

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 Canine Progesterone Reader is a portable fluorescence scanning instrument for measuring the concentration of target analyzes in canine blood, urine and other specimens which will be properly processed in accordance with appropriate Immunoassay Test procedures.

It is used as an aid in the screening of determination of the cause of infertility, track ovulation, diagnose an ectopic or failing pregnancy, monitor the health of a pregnancy.

Progesterone also known as P4 (Pregn-4-ene-3,20-Dione) is a C-21 Steroid hormone involved and the bitch heat cycle, Pregnancy (supports gestation) and embryo genesis of different species. Progesterone belongs to a class of hormones called progestogens, and is the major naturally occurring progestogen.

In mammals, progesterone, like all other steroid hormones, is synthesized from pregnenolone, which in turn is derived from Cholesterol.

Dog Progesterone level Chart

Canine P4 Progesterone Level Chart

Testing Kits and Progesterone Test For Dogs

Progesterone is essential for the regulation of the normal females reproductive functions. The Major physiological actions of progesterone are : a) in the uterus and ovary b) in the mammary gland: lubular-alveolar development in preparation for milk secretion; c) in the brain: neurobehavioral expression associated with sexual responsiveness and d) in the bone: prevent of bone loss.

During the follicular phase of the heat cycle, progesterone levels remain low following the LH surge and ovulation, luteal cells in the ruptured follicle produce progesterone in response to LH. During this, the luteal phase, progesterone transforms estrogen-primed endometrium from a proliferation to a secretory state. If conception occurs, the levels of progesterone are maintained at mid-luteal levels by the corpus luteum until about week six. At the time the placenta becomes the main source of progesterone and levels rise from approximately 50-280 ng/mL.

  • Improper timing of breeding  is often the cause of unsuccessful breeding, so progesterone test is always a good way to go specially if you have problem bitches or ones that will not accept the male.
  • The conception rate after a single breeding is highest if the bitch is bred 3-10 days prior to Day 1 of Di-estrus (D1).
  • After a single breeding, the number of live pups per CL is highest when the breeding occurs 4 days prior to D1 by a proven male. This would equate to the fifth day of estrus for an average, normal 9 day estrus.

Fertile male

The breeding male should be free of canine Brucellocis.

The ejaculate should contain at least 200 x 106 total cells after centrifuging is done

It should have 80 % motility

80% of the cells should have normal morphology.

Dog Breeding

If natural breeding on your bitch is performed, be active in the breeding process of your dogs. Observe the breeding to see if a tie occurred.

Optimal Progesterone level and and timing for optimal breeding.

This is important because there are several reasons why breeding may not occur, or the fertile period may be missed.

Some reasons include: male – female incompatibility caused by physical problems in either the male or female.

Psychological problems.

Behavioral problems.

If there are constraints on the availability of the male, or the number of breeding’s possible, it is important that the cycle be monitored carefully to ensure the optimal time to breed.

Management of breeding’s using shipped, chilled semen or frozen semen require excellent timing of the fertile period.

Hormone assay for either progesterone or LH.

While vaginal cytology is fairly reliable, all other methods are unreliable except for hormone assay.

Hormone assays are the best way to determine the fertile period.

Relative progesterone concentrations during the proestrual/estrual period.

P4 (ng/ml) 

Relative time points

1.1-1.9 Before LH surge

2.1-2.9 Day of LH surge

3.1-3.9 Days 1-2 after the LH surge

4.0-8.0 Day of ovulation

10-80 2-3 weeks post-ovulation

Progesterone Testing

The initial progesterone rise during estrus in the bitch coincides with the LH peak.

Before the LH peak the progesterone is < 1.0 ng/ml. The P4 reads from 1.4 ng/ml

On the day of the LH peak, progesterone rises to around 1.5 – 2.0 ng/ml. Thereafter the progesterone is > 5 ng/ml.

Using the reader ovulation occurs at about 3.8-5.0 ng/ml (11.7 nmol/L) of progesterone (including the day of ovulation and the 24 hours it takes to ovulate) and the best days to breed is when they are about 17.6-24.1 ng/ml (54-74 nmol/L) of progesterone.

Progesterone levels and calculating more or less the day to start.

First of all the blood should be taken from the jugular vein, more of less 6 ML and put in a red cap blood container.

The normal early readings are: .1 – .2 .3 on normal machines the P4 reads from 1.4 ng/mL

Start to draw blood from your English bulldog bitch on the 7th day.

They can go back down to .2 or even .1 – this depends on the time of day the blood was drawn.

When the LH surge goes over 1.0 to 1.9 – you can breed your English bulldog in 5 to 6 days.

If the 5th day seems a little early – you would want to breed your bitch the 6th and 7th day – then skip a day and breed again.

If you draw blood and the level is .2 or .3 – then wait 3 days and draw blood again. 1.4

If you draw blood and the level is .7 or .8 – then you do it every other day

Once you start to see the level at:

2.0 to 2.9 ng/ml = breed in 4 to 5 days

3.0 to 3.9 ng/ml = breed in 3 to 4 days

4.0 to 8 .0 ng/ml = breed today or tomorrow – this is where the LH Surge Occurs.

Over 8.0 ng/ml with no white sells – then the bitch has ovulated

When you hit the level above 4.0 ng/mL this is day 1 of ovulation. Then it takes approximately 48 hours for the ova’s to mature enough to be fertilized…..then they can be fertilized for approximately another 48 hours.

When you breed – you breed 2 days in a row – then skip a day and breed another time.

When you have an English bulldog bitch that do not swell up or has a lot of discharge – then check their thyroid level, and they may have to go on thyroid pills for the rest of her life “Levothyroxine“ is the pill you would need to use.

So in my opinion and Conclusion on progesterone levels after many years of breeding here is what I do and I have a 95% conception rate.

If your bitch is at a level of progesterone of 5 ng/ml and you have good semen from a good stud you can start to breed her and semen should stay alive inside her for at least 5 to 7 days.

If your have semen that was shipped to you and it’s chilled you can start to breed when the progesterone level is at 8.0ng /ML to 12.0 ng/ ML semen should live at least 2 to 4 days inside her.

If you have frozen semen or from an older stud that the motility and quality is no good than you breed when the progesterone level hits at 17 ng/ml to 24ng ml that is when the eggs are fertile and semen will immediately enter the egg.

Canine Progesterone level Chart

Day Estrogen Level Progesterone Level Luteinizing Hormone (LH) Level When To Breed Based On Semen Type Whelping Indicators
-3 day High < 1 ng/ml Low
-2 day Moderate 2-3 ng/ml Surge
-1 day Low 3-4 ng/ml Low
0 (ovulation) Low 5 ng/ml Low
+1 day > 5 ng/ml Natural semen
+2 day > 5 ng/ml Fresh chilled semen
+3 day > 5 ng/ml Frozen semen
+2-3 weeks 10-80 ng/ml
+62 day +/- 1 2 ng/ml Temperature drops
+63 +/- 1 1 ng/ml Whelping
   < = Less than        > = Greater than

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