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Getting started with frozen semen

Ask questions, and get some insight into frozen semen, it's preparation and use.

Moderator: Jos

Getting started with frozen semen

Postby koolkas2 » Thu Aug 30, 2012 1:27 pm

Hello,

Shipping fresh cooled semen has besome a huge problem this year in my area, to a point where I’m considering buying a liquid nitrogen tank and start breeding frozen next year.

I’m just unsure of which size of tank I will need for up to 10-12 mares bred/year, considering I might offer storage to a couple other small breeders? I would look into frozen with LFG contracts and would use a timed AI protocol with 2 doses/cycle if it helps assessing the capacity needed… I’m looking at the MVE tanks. What else should I look into to get started? Canes/goblets, etc?

Thanks!
koolkas2
Neonate
 
Posts: 9
Joined: Tue Jun 26, 2012 9:52 am

Re: Getting started with frozen semen

Postby Jos » Thu Aug 30, 2012 6:00 pm

For a small breeder, the XC Millenium 20 will probably prove to be adequate. This has 6 canisters and good duration of holding time (which means that nitrogen costs are kept at a minimum). One will typically dedicate a canister to each separate stallion (i.e. not mix stallions in a canister) in order to avoid mixing up semen ( :shock: ), although one can put dividers into the canister to make 2 or 4 divisions (although 2 is probably the most in a 20L tank, which has smaller diameter canisters). We have found the best prices and service to be from SemenTanks.com
Image
I will add one "don't" as far as a primary tank is concerned, and that is don't purchase a cheap or used tank (OK, maybe that counts as 2 don'ts!!). All tanks can fail, but the older they are or the cheaper they are, the more likely failure is. There are some cheap imported tanks available that have thinner metal, and the vacuum between the inner and outer layers of the tank is your insulation that keeps your nitrogen (and therefore the semen!) cold. If that vacuum is lost, so is your tank and semen!! It's fine to have an old or cheap tank as a backup tank or a dewar for moving notrogen or semen, but I would recommend a new brand-name (such as MVE) tank be used for your primary tank.

Another don't is don't keep your tank on concrete. Doing so will speed up the possibility of oxidation of the tank (they're made of aluminum, so they don't "rust", but, like steel, aluminum will oxidize, so think of it as "aluminum rust"). Obviously that will then speed up the possibility of vacuum failure and tank/semen loss. A "do" as far as your tank is concerned is do keep it in a place where there is high traffic - somewhere that you walk by every day. This will (a remind you to dip the tank for nitrogen levels (see below); and (b) increase the chances of seeing a tank "frosting up" on the outside, which indicates tank failure (and therefore may give you enough time to get the semen into another tank).

Dip your tanks weekly and record the resulting depth. You should see a predictable decline. If you see a sudden increase, that suggests a problem with the tank. You should refill your tank when there is between one-quarter and one-half LN2 remaining. Theoretically one can go as low as 1" of nitrogen remaining and not damage semen, but I wouldn't recommend that!!! :shock: To dip your tanks, you will require a dipstick. This can be a plastic one purchased for the function from SemenTanks.com or whoever, or you can get a wooden yardstick from the hardware store and paint one side black. The purpose-made one is easier, but the home-made one is cheaper! :D

Often semen will be shipped in canes and goblets already, but it is useful to have a bag of spares on hand in case the straws come "bulk". They are cheap, so not a huge expense there!

A Styrofoam cooler box is useful to have, as that will allow you to pour nitrogen into it and then dump semen straws in there for sorting if needed. You will also need hemostats or forceps to handle the straws with.

To thaw the semen, you will need a suitable container for the warm water, a thermometer to get the water temperature correct, and a timer.

For the insemination you will require syringes and pipettes. If using the half-ml straws, the "Universal" pipette from Minitube is useful, for which you will also require a stylette (which is reusable).

I'm not sure where you are located, but if in the USA, adding P&E or Biorelease Altrenogest to your program will make your timing a little easier. These products are available from BET Pharmacy in Lexington, KY. You will require a prescription for them.
We're always happy to try and help, but don't forget to check the articles section
of the website too, which has a search engine to help you look for answers!
:)
User avatar
Jos
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Re: Getting started with frozen semen

Postby koolkas2 » Thu Aug 30, 2012 8:25 pm

Wow, thanks for all these tips! The XC Millenium 20 is actually the one I was looking at, and found that sementanks.com had very good prices. An older tank as a backup and for nitrogen moving is a good idea! And I wouldn't have thought it was possible to plunge the straws in a styrofoam cooler filled with nitrogen to sort them! Are the straws always marked with the stallion info when you receive them?

If one canister is used per stallion, it means the tank would only be used for 6 stallions?! The 5ml straws are stored directly in the canister, right? I guess one can use canes/goblets marked with the stallion info if using .5ml straws, so it's possible to put more than one stallion in a canister? Might be a silly question, but if 2 goblets are used per cane, when the upper goblet is empty and removed, won't the straws in the lower goblet float out of it?

I do my own AI with fresh cooled and would like to do my own frozen AI once I'm confident handling and thawing the straws. Once thawed, do you empty the straws (.5 or 5ml) and mix the dose to draw it into an insemination pipette using an all plastic syringe? Is it easier/better to use a pipette with stylette?

Sorry, many questions here!

PS I'm in Canada :)
koolkas2
Neonate
 
Posts: 9
Joined: Tue Jun 26, 2012 9:52 am

Re: Getting started with frozen semen

Postby Jos » Fri Aug 31, 2012 11:08 am

Straws should be marked with the stallion's name and if frozen by a reputable facility will be. We have however received straws that were not labelled... (and it surprised us not at all when the post-thaw motility was in the single digits...) :(

One only needs to pour about 3 inches of nitrogen into the Styrofoam cooler - and make sure you do use a Styrofoam one, a plastic one will become brittle and may break! The thicker the cooler the better, as if it's a thin one, it will eventually freeze whatever is underneath it (so don't put it on the linoleum floor - and don't ask me how I know this either! :mrgreen: ).

Yes, 5 ml straws are stored directly in a canister. It is important therefore not to mix different stallions in that canister, although if you were to decant the straw to the Styrofoam box prior to thawing you could check for sure who it is - but it's better not to handle the straws more than one has to. With the 0.5 ml straws in canes/goblets, yes one can mix, but make sure you label the tabs on the top of the cane to make it easier. It is of course still better not to mix in a canister if you can avoid it...

Either try to draw 0.5 ml straws from the bottom goblet first, or else make sure that you leave the top goblet in place even when empty, as yes, the bottom straws may float out of the goblet if you don't.

If you use the Minitube insemination pipette I mentioned above, you do not need to pool semen as you can inseminate individual straws without removing the pipette from the cervix/uterus. If you are not using that pipette, then typically one would pool the semen, draw up into a pipette/syringe and go from there. If you are doing the inseminations by yourself, pooling the semen is easier. If you have helpers, using the Minitube pipette is easier.

As you are in Canada, you will be restricted to using daily-dose P&E, as the biorelease products from BET Pharmacy are not available there.
We're always happy to try and help, but don't forget to check the articles section
of the website too, which has a search engine to help you look for answers!
:)
User avatar
Jos
Senior Stallion or Mare
 
Posts: 3948
Joined: Sun Jun 10, 2012 12:11 am

Re: Getting started with frozen semen

Postby lotsofponies » Sun Oct 21, 2012 11:30 am

How many 5 ml straws will fit in a canister of the XC Millenium 20 ? And how much liquid nitrogen would be in the tank for it to be considered full ? I guess I'm asking do you fill it to the neck or half way up the body of the contain to consider it to be full?
lotsofponies
Weanling
 
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Joined: Mon Jun 18, 2012 1:10 pm

Re: Getting started with frozen semen

Postby Jos » Sun Oct 21, 2012 2:05 pm

Each canister will hold approximately 30 straws, depending upon how tightly one packs them. And the tank holds about 20.5 litres - and it's filled to the neck to be considered full.
We're always happy to try and help, but don't forget to check the articles section
of the website too, which has a search engine to help you look for answers!
:)
User avatar
Jos
Senior Stallion or Mare
 
Posts: 3948
Joined: Sun Jun 10, 2012 12:11 am


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