Abstract
Difructose dianhydride IV (DFA-IV) is produced from levan, which is a natural polysaccharide that belongs to the fructan family, through the activity of levan fructotransferase (LF) derived from microorganisms. Recently, DFA-IV has been expected to have diverse applications in the food and medical industry. Here, we examined the potential application of DFA-IV forin vitro fertilization (IVF) in pigs. In the assessment of acrosomal integrity during incubation, intact acrosomal or viable spermatozoa were highly sustained in 0.1% or 0.25% DFA-IV (69.8%-70.8%,P<0.05). Reactive oxygen species (ROS) levels during sperm incubation decreased following the addition of DFA-IV, and 0.1%-0.5% DFA-IV in particular significantly decreased ROS production relative to that seen with no addition or 0.75% DFA-IV. Total fertilization (mono+ polyspermic oocyte) rate was significantly higher in the addition of 0.1% DFA-IV (94.2%) than with other concentrations (71.8%-86.7%,P<0.05). When using reduced IVF times and lower sperm numbers, we found that addition of 0.1%–0.5% DFA-IV significantly increased the fertilization rate (P<0.05). Fertilized oocytes treated with 0.1% DFA-IV exhibited higher embryonic development and blastocyst formation than those treated with other concentrations (P<0.05). Consequently, the addition of DFA-IV during IVF improved fertilization and embryonic development, suggesting the possible use of novel sugars for enhancement of assisted reproductive technology (ART) in mammals.
Highlights
Sugar is an essential component of the sperm extender because it provides an energy source for sperm viability and maintains the osmotic pressure of the freezing diluent[1,2]
In this study, we examined the potential application of Difructose anhydride (DFA)-IV during in vitro fertilization (IVF) to examine whether this carbohydrate could be used as a new alternative sugar in assisted reproductive technology (ART) in mammals
The broad O–H stretching of DFA-IV was observed around 3,415 cm–1, and weak C–H vibration was observed at 2901 cm–1[16]
Summary
Sugar is an essential component of the sperm extender because it provides an energy source for sperm viability and maintains the osmotic pressure of the freezing diluent[1,2]. 10 January 2017, Epub 20 April 2017 CLC number: R715, Document code: A The author reported no conflict of interest Chemical synthesis of DFA from natural fructans has been shown to have low reaction selectivity, and the product is difficult to separate and purify. Such chemical synthesis causes environmental pollution; enzymatic synthesis using microbes or plants has been considered as a favorable alternative for the preparation of DFA[11]. Levan has been reported to have many applications, the physiologic function of DFA-IV has not been extensively studied to date
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