Publications by authors named "Brian D Whitaker"

4 Publications

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Cyanidin improves oocyte maturation and the in vitro production of pig embryos.

In Vitro Cell Dev Biol Anim 2020 Aug 4;56(7):577-584. Epub 2020 Aug 4.

Department of Animal and Pre-veterinary Studies, University of Findlay, 1000 North Main Street, Findlay, OH, 45840, USA.

The objective of this study was to reduce the negative effects of oxidative stress by decreasing the levels of reactive oxygen species (ROS) through supplementation of the major antioxidants present in elderberries: kuromanin and cyanidin. Oocytes (n = 1150) were supplemented with 100 or 200 μM of kuromanin or cyanidin during maturation, and then evaluated for ROS levels or fertilized and evaluated for penetration, polyspermic penetration, male pronucleus formation, and embryonic development. The ROS levels and incidence of polyspermic penetration were lower (P < 0.05) in oocytes supplemented with 100 μM cyanidin when compared with other treatments. Supplementation of 100 μM cyanidin increased (P < 0.05) MPN and blastocyst formation compared with other treatments. However, supplementation of 100 μM kuromanin did not have significant effects on the criteria evaluated, and supplementation of 200 μM kuromanin had significant (P < 0.05) detrimental effects for each criterion. Additional oocytes (n = 1438) were supplemented with 100 μM cyanidin during maturation and evaluated for glutathione, glutathione peroxidase, catalase, and superoxide dismutase activity. Supplementation of 100 μM cyanidin increased (P < 0.05) catalase activity and intracellular GSH levels compared with no supplementation of cyanidin. These results indicate that supplementing cyanidin during maturation reduces oxidative stress by reducing ROS levels and increasing GSH concentrations within the oocyte.
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http://dx.doi.org/10.1007/s11626-020-00485-yDOI Listing
August 2020

Melatonin and tannic acid supplementation improve fertilization and embryonic development in pigs.

Anim Reprod 2018 Aug 16;15(2):118-123. Epub 2018 Aug 16.

Department of Animal and Pre-veterinary Studies, University of Findlay, Findlay OH, 45840, USA.

The objective of this study was to determine the effects of melatonin supplementation during maturation and tannic acid supplementation during IVF on fertilization kinetics and early embryonic development. Experiment 1 determined the optimum concentration of melatonin supplemented to the oocytes for subsequent embryonic development. Oocytes (n = 400) were supplemented at 22 h of maturation with 0, 75, 100, or 150 nm melatonin and then subjected to IVF and embryo culture. After IVF, a portion of the embryos were evaluated for penetration, polyspermy, and male pronuclear (MPN) formation rates. Embryos were evaluated 48 h after IVF for cleavage and 144 h for blastocyst formation. There were no significant differences between treatment groups with respect to penetration and polyspermy. Supplementation of 150 nm melatonin produced a significantly greater (P < 0.05) percent of embryos with MPN compared to those supplemented with 75 nm or 100 nm. Supplementation of 150 nm melatonin produced significantly less (P < 0.05) embryos cleaved by 48 h after IVF while 75 nm melatonin supplementation had a significantly higher (P < 0.05) percentage of blastocyst formation by 144 h after IVF. Based on the optimal concentration of melatonin observed in experiment 1, experiment 2 determined the effects of supplementing 75 nm melatonin to the maturation media and 5.0 μg/ml tannic acid supplementation during IVF on oxidative stress, fertilization kinetics, and embryonic development. Oocytes (n = 720) were supplemented at 22 h of maturation with or without 75 nm melatonin and then fertilized with frozen-thawed sperm supplemented with or without 5 μg/ml tannic acid. Reactive oxygen species levels were measured in matured oocytes using 2',7'-dichlorodihydrofluorescein diacetate. Oocytes supplemented with 75 nm melatonin had significantly less (P < 0.05) reactive oxygen species generation and oocytes fertilized with sperm incubated with tannic acid had a significantly less (P < 0.05) incidence of polyspermic penetration compared to no supplementation. All treatment groups had significantly greater (P < 0.05) incidence of male pronuclear formation compared to oocytes not supplemented with melatonin and fertilized without tannic acid. Oocytes that were supplemented with melatonin and fertilized with sperm incubated with tannic acid had a significantly higher (P < 0.05) percentage of blastocyst formation by 144 h post-IVF compared all other treatment groups. Results indicate that supplementation of 75 nm melatonin during oocyte maturation and 5 μg/ml tannic acid during IVF leads to a decrease in oxidative stress, increase in IVF success and subsequent embryo development in pigs.
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http://dx.doi.org/10.21451/1984-3143-AR2016-937DOI Listing
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC8186873PMC
August 2018

Effects of anti-lipid peroxidases on frozen-thawed boar spermatozoa.

In Vitro Cell Dev Biol Anim 2011 Jun 13;47(5-6):350-4. Epub 2011 Apr 13.

Department of Biology, Ferrum College, Ferrum, VA 24088, USA.

This study evaluated the effects of anti-lipid peroxidases when supplemented to the thawing and incubation media of frozen-thawed boar spermatozoa. Semen pellets were thawed and incubated in media with 1.0 mM α-tocopherol or diethylenetriamine. After 1 h, the acrosome reaction was induced using calcium ionophore A23187, and acrosomes were evaluated using Wells--Awa staining. The number of spermatozoa with fragmented DNA was evaluated using silver staining after single-cell gel electrophoresis. Membrane lipid peroxidation was measured by the end point generation of malondialdehyde. The diethylenetriamine-supplemented media had a higher (P < 0.05) percentage of acrosome-reacted spermatozoa (84.4 ± 4.1%) compared to the control (78.3 ± 4.2%) and α-tocopherol-supplemented (78.0 ± 3.9%). The control had a higher (P < 0.05) percentage of spermatozoa with fragmented DNA (59.3 ± 4.3%) compared to the DETA (28.7 ± 4.1%) and α-tocopherol supplementation (28.0 ± 3.8%). Spermatozoa supplemented with diethylenetriamine had higher amounts (P < 0.05) of malondialdehyde generated (3.60 ± 0.05 μM/10(7) cells) compared to the α-tocopherol supplementation (0.14 ± 0.05 μM/10(7) cells) and the control (0.12 ± 0.05 μM/10(7) cells). These results indicate that supplementing with either 1.0 mM diethylenetriamine or α-tocopherol during semen thawing and incubation protects against DNA fragmentation, and diethylenetriamine increases the percent of spermatozoa capable of completing the acrosome reaction that could induce membrane lipid peroxidation.
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http://dx.doi.org/10.1007/s11626-011-9403-xDOI Listing
June 2011

Exogenous gamma-glutamyl cycle compounds supplemented to in vitro maturation medium influence in vitro fertilization, culture, and viability parameters of porcine oocytes and embryos.

Theriogenology 2004 Jul;62(1-2):311-22

Department of Animal and Poultry Sciences, Virginia Polytechnic Institute and State University, Blacksburg, VA 24060-0306, USA.

High concentrations of intracellular glutathione (GSH) enhance in vitro production of porcine embryos. Objectives were: (1) to determine the effects of gamma-glutamyl cycle compound supplements to the IVM medium on IVF and IVC; and (2) to evaluate embryo viability. Porcine oocytes were matured in NCSU 23 medium supplemented with either l-cysteine (3.3 mM), l-cysteamine (150 P < 0.05microM), l-cysteine and l-cystemaine, l-glycine (1, 2.5, or 5 mM), l-glutamate (1, 2.5, or 5 mM), l-alpha-aminobutyrate (3.3mM), beta-mercaptoethanol (BME) (25 microM), l-cysteine and BME, or l-alpha-aminobutyrate and BME. Increases (P < 0.05) in GSH concentrations were observed using l-cysteine, 1.0 mM l-glutamate, l-alpha-aminobutyrate, and l-alpha-aminobutyrate with BME. Oocytes matured with l-alpha-aminobutyrate and BME had a lower (P < 0.05) occurrence of polyspermy during IVF compared to controls and a greater percentage (P < 0.05) of embryos reaching the blastocyst stage compared to other treatment groups. For Objective 2, oocytes were matured in NCSU 23 or NCSU 23 supplemented with l-alpha-aminobutyrate with BME. Embryo cell death was determined using an Annexin V-FITC assay. Supplementation had no effect on the time of cell death. Embryo mortality was increased (P < 0.05) from 24 to 42 h post-IVF, with the greatest occurrence around 36 h. In conclusion, supplementing l-alpha-aminobutyrate and BME into the IVM medium increased intracellular GSH concentrations, decreased the occurrence of polyspermy during IVF, and increased embryo development parameters during IVC, but did not affect cell death during embryo development. The onset of cell death occurred from 24 to 42 h post-IVF, with the greatest occurrence around 36 h post-IVF.
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http://dx.doi.org/10.1016/j.theriogenology.2003.10.014DOI Listing
July 2004
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