Basics of sperm function and sperm functional tests; Sperm functionality as it relates to challenges in the female reproductive system; Which tests? Sperm mucous penetration, hypo-osmotic swelling test, hyperactivation, acrosome reaction, assessment of centrioles; Do these tests inform us about potential male fertility? By Gerhard van der Horst (PhD, PhD) and Liana Maree (PhD).
Thursday 1 July 2021.
Webinar #11: Do sperm functional tests inform us about potential fertility in humans?microptic2021-07-01T13:43:37+02:00
Dear colleagues
Now i have listened the 11 th webbinar by Prof Van der Horst. I would like to know, the penetration test using seminal plasma and it it goes on small ruminants to evaluate sperm functionality.
2- Hosh test also evaluate leve sperm. I don´t hear, because soun is low.
3- Acrossome reacted. I didn´t understand, the number of 40 % cover.
Best regards
João Pedro Barbas
(EZN/Portugal)
jpbarbas@gmail.com
1- I would like to know, the penetration test using seminal plasma and if it it goes on small ruminants to evaluate sperm functionality. What functionalities tests did you suggest for small ruminants functionality semen evaluation.?
Below professor Gerhard van der Horst’s answer:
Yes in principle sperm mucous penetration can be done in domestic animals but remember two crucial issues.
1st- The concentration of sperm in semen of for example ruminants is extremely high. Accordingly no CASA can be done like with humans where we analyse sperm movement in semen without dilution. Accordingly one has to make a modification for domestic animals. Perform Differential centrifugation like 45% to 90% PureSperm or equivalent with half of the semen. After centrifugation only clean (no sperm) seminal plasma will remain on top. Now dilute original semen (other half of sample) with clean seminal plasma so that the sperm concentration is less than 25 million/mL. The analyse normal CASA.mot
2nd- One now has to develop cut-off points in terms of those kinematics used for human but of course with ROC curves or with frequency diagrammes to determine the new values for a particular species or breed. The reason is that the values are not known. Another good reason to do this is that many believe that for cryopreservation in animals like rams the seminal plasma may provide additional protein support during freezing and so it is good that this dilution is done and also the values known in seminal plasma.
2- Hosh test also evaluate leve sperm?. I don´t hear, because sound of webinar is somewhat low.
I suppose you mean HOS test. Indeed, this test is used in order to study the integrity and functionality of the sperm membranes, so according to this, all sperms that react to the hypoosmotic solution (they roll their tails) are considered alive. In contrast, sperms with straight tail are dead (please find more information in WHO 5th edition manual). One of the advantages of the stain we use for vitality analysis, called BrightVit, is that it combines eosin-nigrosine technique together with HOS test, a you see in the image shown during the webinar. So it is possible to study both the staining of sperm heads (red/white) and the tail rolling.
3- Acrosome reacted. I didn´t understand, the number of 40 % cover. What means?
Regarding acrosome, according to WHO 5th edition manual, for a sperm to be considered normal, acrosome needs to cover between 40-70% of the sperm head. If it is smaller or bigger it could arise some problems.
We hope we have solved your doubts