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Title:The combinative effect of soluble suLeA glycan and specialized bovine semen extender, FSRD4+, on bull sperm quality
Author(s):Messer, Alex
Advisor(s):Miller, David J.
Contributor(s):Bahr, Janice; Knox, Robert
Department / Program:Animal Sciences
Discipline:Animal Sciences
Degree Granting Institution:University of Illinois at Urbana-Champaign
Degree:M.S.
Genre:Thesis
Subject(s):Glycans
suLeA
suLeX
Bull Sperm
FSRD4+
Motility
Viability
Semen Extender
Abstract:Artificial insemination is an invaluable tool that has been adopted by the cattle industry, primarily dairy, and continues to be a crucial reproductive technique. Likewise, cryopreservation is ubiquitously used in the U.S. as the preferred method to store bovine sperm due to convenience despite damage to sperm during processing and thawing. For this reason, one of our objectives was to determine if a liquid extender, FSRD4+, could extend the lifespan of sperm by days. Since longevity with minimal sperm damage was an ultimate goal, we also tested the effect on longevity by adding oviduct glycans to medium and extender. Before fertilization and while bound to the oviduct, sperm become quiescent and have prolonged viability and maintained motility. Additionally, sperm undergo capacitation to reach final maturation. Capacitation will allow the sperm to hyperactivate and penetrate the cumulus cells and the zona pellucida, and fertilize the oocyte. The sperm-oviduct binding and subsequent sperm reservoir formation is facilitated by glycan-lectin binding. In cattle, the Lewis A trisaccharide (LeA) in the oviduct binds sperm. We tested sulfated LeA, because it binds sperm with higher affinity, for its effects on sperm viability and motility compared to controls. Four variables were applied: temperature (5°C, 15°C, 22°C, 37°C), glycans (suLeA, suLeX, no glycan), medium (FSRD4+ and dmTALPC), and time (1, 24, 48, 72, 96, and 120 hr). Semen was collected and shipped overnight for experiments. Sperm were washed, incubated in their respective temperature, glycan, and media treatments, and analyzed for viability and motility. Viability was determined using fluorescent probes SYBR14 and propidium iodide (PI). Motility was determined as total motility where progressive and non-progressive motility were summed. Significance was determined by a p-value of ≤ 0.05. The results indicated viability had improved significantly in FSRD4+ extender had significantly higher viability than dmTALPC, both suLeA and suLeX performed similarly but had significantly higher viability than no-glycan treatment, and 15˚C and 22˚C had significantly higher viability than 5˚C and 37˚C. Similarly, sperm motility was significantly higher in FSRD4+ extender compared to dmTALPC. Sperm motility was higher with suLeX compared to suLeA and no-glycan, and suLeA compared to no-glycan. Over the 120 hr, 15°C storage yielded significantly more viable, motile sperm than 5˚C, 22°C, and 37˚C, despite it having significantly lower motility than 22°C and 37˚C at the 1 hr time point. To summarize, sperm in FSRD4+ extender, with suLeA, and at 15°C or 22°C had significantly higher viability, and sperm in FSRD4+ extender, with suLeX or suLeA, and at 15˚C had significantly higher motility compared to other treatments. In conclusion, FSRD4+ and soluble glycans significantly improved sperm viability and motility during long-term liquid storage with the best combination of medium, glycan, and temperature as FSRD4+ extender, suLeA, and 15˚C.
Issue Date:2018-11-27
Type:Text
URI:http://hdl.handle.net/2142/102807
Rights Information:Copyright 2018 Alex Messer
Date Available in IDEALS:2019-02-07
Date Deposited:2018-12


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