Abstract
The aim of our research was to examine the ability of density gradient preparation BoviPure® and swim up method on bull sperm separation and in vitro embryo production (IVP) systems. Frozen/thawed semen from six Simmental bulls was pooled and treated using both methods. The sperm motility, concentration, membrane activity, membrane integrity and acrosomal status were evaluated and compared before and after sperm processing using BoviPure® and swim up methods. We also evaluated and compared cleavage rates, embryo yield and quality between the methods. There were significant differences (P < 0.05) between the sperm characteristics before and after BoviPure®, but not after swim up method. However, there were significant differences for sperm results among those two mentioned methods. A total of 641 oocytes were matured and fertilized in vitro and cultured in SOFaaBSA. The percentage of cleavage (Day 2) and the percentage of hatched embryos (Day 9) were similar for both methods. However, embryo production rate (Day 7) was significantly higher using BoviPure® method (P < 0.05). Also, total cell number and embryo differential staining (inner cell mass and trophectoderm cells) of Day 7 morulas and blastocysts showed that BoviPure® treated sperm displayed higher quality embryos compared to swim up method (P < 0.05). Our results indicate that BoviPure® method has an enhanced capacity in sperm selection for in vitro embryo production when compared with swim up method. So, we concluded that BoviPure® could be considered as a better alternative to swim up method for separating bull spermatozoa from frozen/thawed semen for IVP of bovine embryos.
Highlights
Mammal spermatozoa have very expressive heterogeny in morphology, motility and nuclear stability
Total cell number and embryo differential staining of Day 7 blastocysts showed that BoviPure® treated sperm displayed better quality embryos compared to swim up method (P < 0.05)
Membrane activity and acrosome status evaluated by hypoosmotic swelling test (HOS), SYBR-14/propidium iodide (PI) and EthD-1/FITC-PSA tests showed a higher percentage of live and acrosome intact spermatozoa obtained after BoviPure® method compared to swim up
Summary
Mammal spermatozoa have very expressive heterogeny in morphology, motility and nuclear stability. Cervical mucus represents a barrier which allows only migration of progressively motile spermatozoa with normal morphology and high nuclear stability [1]. Frozen bull spermatozoa after thawing have lower percentage of progressive motility (30–70%), but percentage of morphologically normal spermatozoa in thawed ejaculate is equal to fresh semen [2]. Sperm separation procedures are able to significantly improve the sperm quality with higher rate of progressive motility and morphologically normal spermatozoa. In the in vitro production of embryos, sperm separation methods have very important role. Such selection of spermatozoa separates motile sperm from nonmotile, removes seminal plasma, cryo-
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