A novel flush technique to simulate natural dispersal of spermatozoa in the female reproductive tract and expedite motility assessment of fresh ejaculated Merino (<i>Ovis aries</i>) sperm

Abstract

Motility is an important criterion in the assessment and quantification of the quality of ejaculated and epididymal sperm samples. Ovine sperm spermatozoa are particularly susceptible to cryodamage, and shortening the interval from collection to cryostorage may potentially minimize the negative effects of cryopreservation, thereby improving the post-thaw viability of ram spermatozoa. The use of the swim-up technique (SUT) in quantifying spermatozoa motility is well documented, especially for the isolation of highly motile spermatozoa for assisted reproductive purposes. However, this technique is time consuming and involves a swim-up period of 10 minutes before the motility of a sample is recorded by using computerassisted sperm analysis (CASA) software such as the Sperm Class Analyser® (SCA®). The novel flush technique (FT) allows for capturing of sperm motility tracks via the SCA® system shortly after semen collection, that is, within a minute. This study compared fresh ejaculated sperm motility traits by using the SUT and FT. Motility evaluations were performed using 45 semen samples obtained from 15 adult Merino rams (Ovis aries) at weekly intervals. Motility recordings were captured at 100 frames per second, using a calibrated 20 µm deep Leja slide. The percentage total motile spermatozoa of samples subjected to the FT was 89.2%, which was significantly higher than that recorded by the SUT (83.9%). The results also indicated that the wobble (WOB) parameter showed significantly higher values when using the FT, and parameters curvilinear velocity (VCL) and amplitude of the lateral head displacement (ALH) indicated significantly higher values when using the SUT. Establishing the ideal spermatozoa concentration and analysis of sperm subpopulation motility characteristics would assist in the optimization of the FT, and its use in CASA motility analysis of ovine spermatozoa. Standardization of CASA analysis of ovine semen samples, which would enable the selection of quality spermatozoa samples for use in field insemination (fresh samples) or in vitro fertilization programs, and laparoscopic AI cryopreserved samples warrants further investigation.

Quantitative sperm characteristics of Tankwa goats with special reference to hyperactivated motility

“…Spermatozoa were exposed to four pre-warmed (37 °C) media, namely phosphate buffered saline (PBS) (control), BO sperm wash (containing 10 mM caffeine, as proposed by Brackett and Oliphant (1975)), 4% lignocaine in PBS (G. van der Horst, personal communication, 2016) and 5 mM procaine hydrochloride in PBS (McPartlin et al, 2009). According to the flush technique (Boshoff et al, 2018), 2 µl of each medium was used to flush 1 µl semen into the chambers of a Leja slide (Leja Products B.V., Nieuw-Vennep, The Netherlands), while keeping the slide on a heated stage. Sperm motility was evaluated for each semen sample five minutes after exposure to the media.…”

Aqueous leaf extract ofMoringa oleiferareduced intracellular ROS production, DNA fragmentation and acrosome reaction in Human spermatozoa in vitro

“…In other in vivo studies, however, sperm qualitative characteristics in male rats remained unchanged following treatment with MO (Awodele et al, 2012;El-Sheikh et al, 2016). Sperm functional capacity in mammals depends strictly on their morphology, progressive motility, intact plasma and acrosome membranes (Boshoff et al, 2018). To the best of our knowledge, this is the first scientific report to evaluate the effects of aqueous MO leaf extract on the physiological functions of human spermatozoa in vitro.…”

Origin, Migration, and Reproduction of Indigenous Domestic Animals with Special Reference to Their Sperm Quality

“…Table 4 and Table 5 include macroscopic, microscopic, and some sperm functional characteristics of various indigenous versus exotic breeds of pig, cattle, ram, and goat [ 32 , 33 , 34 , 35 , 36 , 37 , 38 , 39 , 40 ]. By and large, there are many similarities when several sperm parameters of these breeds are compared for a given species.…”