Response to cooling of pony stallion semen selected by glass wool filtration

Abstract

The aim of this study was to compare the sperm separation technique using filtration through glass wool compared with just diluted cooled semen. Eighteen ejaculates were collected from 6 pony stallions of the Brazilian pony breed. Evaluations were done on pH, osmolarity, total motility, membrane functionality (HOST), membrane integrity (CFDA/PI), morphology and mitochondrial viability (MTT) in fresh, 24 and 48 h of cooled semen at 5°C. After dilution, the half of the extended semen was cooled (control group). The other half was cooled after filtration trough glass wool (filtered group). Retained semen was considered the portion of cells that did not transpose glass wool barrier. Total motility from the control, filtered and retained groups after 24 h of cooling was 35.5%, 43.3% and 10% (p < .0001) respectively. Sperm membrane integrity percentage at the CFDA/PI test was 37.9%, 44.8% and 14.8% (p < .0001), on the control, filtered and retained groups respectively. The results confirmed that the passage of spermatozoa through glass wool increased the selection of spermatozoa from pony stallions with higher motility, mitochondrial viability and membrane integrity for cooling in milk extender up to 24 h. Moreover, it was not obtained higher sperm parameters to control after cooling 48 h under the conditions that the study was conducted.

Glass wool column filtration for stallion sperm selection: A comparative analysis with the single‐layer colloid centrifugation

“…This study aimed to assess the efficacy of GWCF as an alternative method to the Androcoll‐E™ centrifugation. The GWCF procedure was selected based on several studies that described its efficacy (Arzondo et al, 2012; Bussalleu et al, 2009; den Van et al, 1997; Hammadeh et al, 2001; Husna et al, 2016; Kim et al, 2010; Lee et al, 2009; Nani & Jeyendran, 2001; Pessoa et al, 2017; Sauer et al, 2012). In the horse, few promising reports were previously published (Casey et al, 1993; Sieme et al, 2003).…”

“…In buffalo semen, GWCF selection resulted in increased sperm motility in the filtrate (Husna et al, 2016). Finally, GWCF using pony semen samples improved motility, as well as mitochondrial and plasma membrane integrity (Pessoa et al, 2017). In addition to these findings, human sperm GWCF was associated with a decrease in DNA damage (Hammadeh et al, 2001; Sauer et al, 2012) and enhanced performance in sperm selection for ICSI procedures when compared to two‐layer density gradient centrifugation (den Van et al, 1997).…”

“…In buffalo semen, GWCF selection resulted in increased sperm motility in the filtrate (Husna et al, 2016). Finally, GWCF using pony semen samples improved motility, as well as mitochondrial and plasma membrane integrity (Pessoa et al, 2017).…”

“…In humans, previous studies showed a higher sperm vitality in suspensions recovered after GWC filtration (GWCF) when compared to pre-GWCF (Nani & Jeyendran, 2001). Later reports described a good sperm quality in boar (Basioura et al, 2021;Bussalleu et al, 2009), canine (Kim et al, 2010), bovine (Arzondo et al, 2012;Lee et al, 2009), buffalo (Husna et al, 2016) and pony (Pessoa et al, 2017) sperm after GWCF. In the study carried out in boar semen, GWCF resulted in improved sperm quality, as evidenced by a higher vitality, morphology and plasma and mitochondrial membranes integrity (Bussalleu et al, 2009).…”

Mammalian sperm capacitation: In vivo and in vitro juxtaposition

“…Also, some parameters such as apoptosis, DNA integrity, membrane maturation, and sperm ultrastructure must be considered notably due to the presence of centrifugation step in both techniques that generate ROS and has a detrimental effect on sperm quality (Oseguera-López et al, 2019). Based on those notions, several advanced methods and natural approaches to sperm selection have enormous potential use to shift from the old methods, such as magnetic-activated cell sorting (MACS) (Gil et al, 2013;Nadalini et al, 2014), hyaluronic acid (HA) binding, Zeta method, and glass wool filtration (Pessoa et al, 2017). For further insight and better understanding of those advanced techniques above, readers are addressed to read some literature reviewed by (Sharma et al, 2015), (Agarwal et al, 2014), and(Oseguera-López et al, 2019).…”