The Effect of Sperm Concentration and Storage Vessel on Quercetin‐Supplemented Rabbit Semen During Chilled Storage

Abstract

Extending the shelf life of chilled rabbit spermatozoa is vital for the expansion of the farmed rabbit industry. This study evaluated the relationship between sperm concentration and packaging on in vitro quality of chilled rabbit semen over 96 h. Semen was collected from adult bucks (n = 4) and pooled at 37°C following evaluation. Pooled ejaculates were diluted with a Tris-based extender supplemented with 100 μm quercetin to a concentration of 15, 30 or 60 × 10(6) spermatozoa/ml, packaged into plastic tubes or 0.5-ml straws and stored at 15°C. Sperm quality was assessed by computer-assisted sperm Analysis [total motility (tMOT)] and flow cytometry [viability, acrosome integrity, H2 O2 production, plasma membrane disorder, apoptosis and DNA fragmentation index (DFI)] at 0, 48, 72 and 96 h. From 48 h, concentrations of 30 and 60 × 10(6) spermatozoa/ml reported the highest tMOT, irrespective of storage vessel (p < 0.05). Storage in straws reduced oxidative stress and improved plasma membrane stability. The %DFI, mean DFI and SD-DFI were increased in spermatozoa stored in tubes compared with straws (p < 0.05). Although the use of low sperm concentrations in artificial insemination doses would facilitate greater dispersion of genetically superior rabbit bucks, dilution to 15 × 10(6) spermatozoa/ml had a detrimental impact on motility. As such, chilled storage at 30 × 10(6) spermatozoa/ml may provide a suitable balance between motility and H2 O2 production to best maintain overall sperm function and should be evaluated in a large-scale AI trial.

Preservability of rabbit semen after chilled storage in tris based extender enriched with different concentrations of Propolis ethanolic extract (PEE)

“…and apeginin-7-glucoside (1.113 mg/g extract) that have scavenging antioxidant activity against free radicals interpret the benefit beyond the enrichment of rabbit semen tris-based extender to overcome the lipid peroxidation process induced via chilling period [54][55][56][57][58][59]. This elongates the period of chilling in rabbit extended semen above 72 h in agreement with Di Lorio et al [9] and Johinke et al [10].…”

Semen supplementation with palmitoleic acid promotes kinematics, microscopic and antioxidative parameters of ram spermatozoa during liquid storage

“…Researches showed that in case of using the ram semen within the short time after collection for artificial insemination, to avoid from freezing/thawing induced injuries, using cooled and diluted semen would be a practical alternative (King et al., ). The improvement in semen storage techniques requires in‐depth knowledge of the biochemical processes occurring during semen collection, dilution and processing (Johinke, de Graaf, & Bathgate, ; Yoshida, ).…”

Supplemented stallion seminal plasma can improve impaired motility due to the dilution effect in chilled Asian elephant sperm

“…In this study, only sperm motility was affected by semen dilution, but not for sperm viability and acrosome integrity. There had been demonstrated in chilled semen that (i) the sperm plasma membrane was more resistant to osmotic stress than the mechanisms responsible for sperm motility, as seen in bull (Liu & Foote, ), boar (Fraser, Gorszczaruk, & Strzezek, ) and dog (Songsasen et al., ); (ii) the sperm viability and acrosome integrity were not differ among different extenders, while the motility was significantly affected, as seen in boar (Kaeoket, Srisowanna, Wichaidit, Chanapiwat, & Manee‐In, ) and donkey (Dorado et al., ); (iii) the sperm plasma membrane was altered after dilution only when final sperm concentration was 20 × 10 6 sperm/ml or less, as observed in bull (Garner et al., ); (iv) the sperm plasma membrane was not affected by semen dilution until 96 hr of cooled storage as shown in rabbit (Johinke et al., ). These indicated that the sperm motility was more susceptible to various conditions, than their plasma membrane.…”

“…A high dilution ratio could result in detrimental effects on semen quality, such as sperm motility, viability or acrosome integrity. This has been demonstrated in boar (Lipensky, Lustykova, Frydrychova, Rozkot, & Vaclavkova, ; Weitze, Le Thi, & Waberski, ), bull (Garner, Thomas, & Allen, ; Garner et al., ), stallion (Hayden et al., ; Varner, Blanchard, Love, Garcia, & Kenney, ), tomcat (Prochowska, Niżański, Ochota, & Partyka, ) and rabbit (Johinke, de Graaf, & Bathgate, ). This might due to the decreased concentration of seminal plasma (SP) (Maxwell & Johnson, ).…”