I want to know the damage that can be caused to the proportion of the compound when the sperm transport from the farm to the laboratory and What is the best way to save it?
Dear GACI,
In my opinion, the routine short-term preservation steps should be applied, as follows:
1- Dilute the semen (within TRIS, skimmed-milk, Na citrate based extenders) at low rates at 26-32 degrees C.
2- Lower the semen temperature further (immediately) down to +4 degrees C by slow cooling (e.g. within a glass material filled with water at room temp. by 2-4 h) and keep at that temp. (within a flask container) till the time of the insemination (within 1-4 days of collection depending on the species concerned).
2- Adding some glycerol, as cryoprotectant, at low concentrations (that may vary depending on the species), if applicable.
3- Adding antioxidants, as appropriate (e.g. Trehalose, SDS, BHT, etc.).
4- Adding some egg yolk (5% or 10%) depending on the species concerned,
5- Adding sugars as energy source (e.g. fructose, mannose, lactose, etc.),
6- Adding Antibiotics (Pen + Strep.),
P.S.; In summer, following almost 150-200 km of transportation at around 30 degrees C (in a pocket of the T-shirt, I transferred the neat ram semen within 1.5-2 h. Then I observed almost 15-20 % dead sperm. I presumed that almost half (10%) of that dead proportion might have been due to transport effect.
That's all.
Regards,
Omer
Hi Gaci,
it depends which species you are working with. You should use a refrigeration extender and dilute it properly (in the horse we recomend at least 1:3 dilution, maximum of 50x10 6 sperm/ml and the final volume being 20 ml for a dose). This depends on the species and largely varies... as does the refrigeration extender.
If you are going to use it immediately (in 30 minutes or so) room temperature should be fine. Otherwise I'd refrigerate it as Omer Ucar suggested. There are some styrofoam boxes in which you can place ice blocks and your doses in a different compartment and will allow for slow cooling and manteinance of the doses at 4 degrees. If you are doing boar semen, then you sholud transport it at 16 C, so you'd need a portable incubator set at this temperature.
If you are using it for experiments, your extender may vary so it does not interfere with your results (if you need your extender devoid of milk, or glucose or calcium lets say...) and as long as the transport is not too long, you could so it at room temperature as well.
I hope this helps,
Bea
dears omer and Beatriz Macias Thank you very much for your attention, your answers convincing.
Dear Gaci, both colleagues are correct. First of all, the species is the most important thing to know the best to maintain the semen quality. Then, is it is going to the lab for standard analysis previous to use in A.I., or for research. In fact, this is important, as saying by Beatriz, because you must assure that there will be non interference between the extender and the tests you are going to do. So, if you give us such information, we can give more advice.
Gaci
first dilute the semen sample,made buffering conditions with respective buffers,antioxidants and sugars, according to your species.Finally put that semen containing ampules in liquid nitrogen container,until that semen is used for further analysis
dear G.K
i got your question . you want to know that how to maintain sperm quality from the site of collection to the site of processing i.e. lab. it depends how much time it takes from the collection site to the lab. if it involves 3-4 hours then you will have to go for initial dilution at the time of collection with a tris based extender with glycerol at 7% level . put the container with diluted semen in a portable refrigerator at 5 degree C and start transporting it to the lab. the time taken in transport shall be counted as equillibration time. which is generally 4 hours for bull semen. if you are working with other species then you may have to change the protocol, extender, cryoprotectant etc. however the time of transport can be managed as equillibration time. i hope this may help you
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