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    Effect of Ascorbic Acid and Proline Amino Acid Supplementations on Cryosurvival and Fertility Rates of Cryopreserved Honey Bee (Apis mellifera) Semen
    (Mary Ann Liebert, Inc, 2022) Guel, Aziz; Yildiz, Cengiz; Cetin, Nurdan Coskun; Yalcin, Oguz Kaan
    This study investigated the effect of ascorbic acid (vitamin C) and proline amino acid alone or together on the quality and fertility of frozen/thawed honey bee spermatozoa. The experiments were designed to compare a single ascorbic acid, a single proline amino acid, and different combinations of ascorbic acid with proline amino acid on the cryopreservation of honey bee semen based on sperm motility, viability, intact membrane (hypo-osmotic swelling test), and fertility rates. Eight cryopreserved study groups comprised Control II with no supplement, along with groups with ascorbic acid (2 mg), proline 25 mM, proline 50 mM, proline 100 mM, and combination groups of both ascorbic acid (2 mg) and proline 25 mM, proline 50 mM, and lastly proline 100 mM groups, respectively. Using 50 mM proline in the tested groups had the greatest impact on sperm motility, viability, the percentage of spermatozoa with intact membrane, and fertility. The cryopreservation process caused a gradual decrease in motility, viability, intact membrane (p < 0.05), and fertility rates (p < 0.01) in all the tested research groups as against the fresh semen control group. Successful honey bee sperm cryopreservation and fertility are achievable when using an appropriate sperm freezing protocol and antioxidant. Proline amino acid as an antioxidant in semen extender had a more beneficial influence on sperm quality parameters and fertility. The success of cryopreservation with antioxidants is related to the chosen antioxidant in a dose-dependent manner.
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    THE EFFECT OF DIFFERENT PRESERVATION MEDIA AND TEMPERATURES ON SPERM QUALITY AND DNA INTEGRITY IN MOUSE CAUDA SPERMATOZOA
    (Cryo Letters, 2022) Yildiz, Cengiz; Coskun Cetin, Nurdan; Yavas, Ilker; Yalcin, Oguz Kaan; Yilmaz, Firdevs; Karaca, Fikret
    BACKGROUND: Mouse sperm can be stored for long or short-time periods. Nevertheless long-term storage leds to significantly reduced sperm quality and fertility because of cryodamage. Thus, in the storage of semen in mice, it is necessary to focus on media and temperatures that gives good results in short-term storage. OBJECTIVE: To determine favorable media for short-term storage of mice spermatozoa by evaluating progressive motility, viability, membrane function integrity, acrosome integrity and fragmented DNA rates at various storage temperatures. MATERIALS AND METHODS: Mouse spermatozoa were collected from epididymides of mature CD1 males and samples were stored at 24 degrees C and 4 degrees C for 60 h. RESULTS: Motility, viability and membrane function of mice spermatozoa were greatest when stored in KSOM media. Motility and viability were not different when stored at refrigerator or room temperature in KSOM compared to HTF or PBS mediums for 48 h, but were after 60 h. There wasn't any significant variation in terms of acrosome integrity in different preservation conditions. Fragmented DNA rates were similar in fresh sperm with KSOM and HTF media, while there was higher damage in PBS medium at 60 h. Overall, sperm parameters were affected significantly by the time of storage and type of preservation medium, and PBS extender was not suitable for mice spermatozoa at room and refrigerated temperatures as it caused the lowest progressive motility, viability, membrane function integrity and the highest DNA damage. CONCLUSION: Mice spermatozoa stored in KSOM retained the best sperm quality parameters both 24 degrees C and 4 degrees C for the first 48 h.