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Help fractionating semen

Equine-Reproduction.com Bulletin Board » General Stallion Questions » Help fractionating semen « Previous Next »


Author Message
 

Worldwide
Neonate
Username: Wweb

Post Number: 1
Registered: 05-2007
Posted on Wednesday, May 16, 2007 - 03:02 pm:   Edit Post Delete Post    Move Post (Moderator/Admin Only)

We recently found out our stallions seminal plasma is toxic to his sperm - thereby making his shipped semen not motile @ 24hrs. A vet did a test ejaculate with only the sperm rich portion and it lived wonderfull. We tried this at home.... This is what we were told - Have 2 cups one for clear and one for the "milky Portion" - The person holding the AV will say "pulsing" when the pulsing begins.
We did this - the collecter switched cups when I said pulsing but upon evaluation it was not very milky in color and the concentration was only 20 m/ml -- we tried again 1 hour later and the collecter switched before I said "pulsing" and we still did not get most of the sperm rich potion - only 60 m/ml (stallion will produce 160-180m/ml on a regular daily collection schedule) Can anyone help with our technique. How much seminal plasma is expelled, can they ejaculate the sperm rich portion before the pulsing is felt? How can we tell when the Gel is being released?
Thank YOu!!
 

Jos
Board Administrator
Username: Admin

Post Number: 1313
Registered: 10-1999
Posted on Wednesday, May 16, 2007 - 04:02 pm:   Edit Post Delete Post    Move Post (Moderator/Admin Only)

An easier technique that we use is that rather than using an open-ended AV, use a disposable liner and make sure that the liner extends beyond the AV far enough to place ones hand easily. This is going to work better with the open-ended AV's such as the INRA (French) AV or the Colorado AV. It can be done with a Missouri, but (as you will understand in a moment) is less reliable.

As the stallion is ejaculating, you should - even in a normal situation - have the glans of the penis close to, at or even past the end of the AV. This, in the case of fractionation, will allow you to observe the ejaculatory spurts.

As the stallion is ejaculating, the person holding the AV, or an assistant, has hold of the section of the disposable liner distal to the AV between the AV and the collection bottle. It is easy to identify each ejaculatory spurt, and after 3 spurts the handler pinches off the disposable sleeve, having allowed the first three spurts to descend into the collection bottle. The subsequent spurts are on the AV side of the handlers "pinch" and are discarded after the stallion has removed his penis from the AV (having been allowed to complete ejaculation!).

Incidentally, you reference the gel in the last line - this is not the same as the seminal plasma. Is the negative response that the sperm are exhibiting in response to the seminal plasma or the gel? If the gel, you have less of an issue, as that is ejaculated right at the end of the ejaculation.
 

Worldwide
Neonate
Username: Wweb

Post Number: 2
Registered: 05-2007
Posted on Wednesday, May 16, 2007 - 10:35 pm:   Edit Post Delete Post    Move Post (Moderator/Admin Only)

We use a Missori AV. - I am SO confused....I was under the impression that an ejaculate has 3 parts - 1)pre ejaculate, including seminal plasma, 2)sperm rich portion, and 3) the gel. If we are collecting the first 3 spurts isnt that the pre-ejaculate which includes the seminal plasma?

How do you determine the spurts in the Missori AV?

Our stallion is usually squirting prior to being deflected into the av, I wait until I feel the pulsing and tell them to collect that portion but it seems the sperm rich portion is coming prior to the pulsing-

I can not tell when the gel portion is being released after the pulsing - will it appear different to the assistant? We got some gel in the second collection but it seemed to easily separate off once the dose was in a syringe it floated to the top and I expelled it from the sample.

Does the pulsing have anything to do with the collecting of fractioned semen?

I am unsure if the Gel or seminal plasma is the bigger problem as when the vet collected it they began by placing one cup at the end of the AV - When the vet felt pulsing she said "pulsing" at which point the assistant switched the cup,- then she said "good" - indicating not to collect the gel. How she knew that with her back to him and just a hand on the base of his penis I have no idea but they got exactly what they needed (only the sperm rich portion) and made it look easy.

I have thought perhaps a centifuge would be easier but the sperm would still come into contact with the seminal plasma.

Thanks for your help - we need to figure out how to do this as it is the ony way we can ship him.

Jen
 

Jos
Board Administrator
Username: Admin

Post Number: 1315
Registered: 10-1999
Posted on Wednesday, May 16, 2007 - 11:36 pm:   Edit Post Delete Post    Move Post (Moderator/Admin Only)

You're right in that an ejaculate comprises of three parts. You're wrong in what those three parts comprise of. :-)
  • Pre-ejaculate - the clear fluid that drips from the stallions penis prior to breeding/collecting;
  • Sperm-rich fraction - self explanatory in content; consists of about the first three ejaculatory spurts (note "spurts" not "drips" as in pre-ejaculate!);
  • Tail-fraction - consists primarily of the secondary sex gland secretions which will include seminal plasma and gel fraction.

Fractionating the ejaculate is more difficult with the Missouri AV - its' one of the reasons we're not really keen on them... You will have to make sure that the stallion's penis is well through the heated lumen of the AV and into the triangular distal end, with the penile opening hard against the end. That way you can (should) see the ejaculatory spurts as they deposit out of the end of the AV. Note that this is in fact pretty much where the stallion's penis should be at the time of ejaculation. If the penis is not clear of the end of the AV there is distinct risk of heat shock to the sperm.

Pulsing at the base of the penis is moderately reliable as an indicator of ejaculation, but I'm guessing you're watching the sperm rich fraction go flying by you onto the ground and then make a point of catching the first part of the tail fraction of the ejaculate... :-)

Fractionation is easy once you get the hang of it, and it's a lot less fiddly than centrifugation for something like this.



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