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Bisphenol AF Caused Reproductive Toxicity in Rats and Cineole Co-Treatment Exhibited Protective Effect
(Springer Heidelberg, 2024) Uyar, Ahmet; Cellat, Mustafa; Kanat, Ozgur; Etyemez, Muhammed; Kutlu, Tuncer; Deveci, Mehmet Zeki Yilmaz; Yavas, Ilker
Bisphenol AF (BPAF) is increasingly used and now found in products intended for human consumption. The protective effect of 1,8-cineole (CIN) against BPAF-induced reproductive toxicity was investigated. Four groups were created, with each group consisting of eight rats: control, BPAF (200 mg/kg), CIN (200 mg/kg), and BPAF + CIN groups. The results demonstrated that the BPAF group exhibited a decline in testosterone levels and a decrease in sperm parameters compared with the control. Additionally, higher levels of MDA were observed, along with lower levels of GSH and GPx activity. CAT activity also decreased slightly. Tnf-alpha, Nf-kappa B levels were significantly higher, and caspase-3 expression was elevated, while PCNA expression decreased. BPAF significantly increased tissue degeneration compared with the control. However, the BPAF + CIN group showed statistically significant improvements in sperm parameters, except for concentration. They also exhibited an increase in testosterone levels and an improvement in MDA and GSH levels compared with the BPAF group. However, GPx activity partially enhanced. Tnf-alpha and Nf-kappa B levels were significantly reduced, and caspase-3 levels declined while PCNA and Bcl-2 levels increased. The Johnsen Testicular Biopsy score showed a substantial increase. Overall, these results suggest that CIN co-treatment in rats enhanced reproductive health and exhibited antioxidant, antiapoptotic, and anti-inflammatory properties against BPAF-induced testicular damage.
Carvacrol showed a curative effect on reproductive toxicity caused by Bisphenol AF via antioxidant, anti-inflammatory and anti-apoptotic properties
(Pergamon-Elsevier Science Ltd, 2023) Uyar, Ahmet; Cellat, Mustafa; Kanat, Ozgur; Etyemez, Muhammed; Kutlu, Tuncer; Deveci, Mehmet Yilmaz Zeki; Yavas, Ilker
Bisphenol AF (BPAF) is an endocrine disruptor, and human exposure to these chemicals is growing in industrialized nations. BPAF has been demonstrated in studies to have toxic effects on reproductive health. This study examined the effects of oral exposure to BPAF on the reproductive system and the protective effects of carvacrol in rats. From 32 Wistar albino rats, four separate groups were set up for this purpose. Carvacrol 75 mg/kg and BPAF 200 mg/kg were administered by oral gavage method. Rat sperm parameters and serum testosterone levels were measured after 28 days of administration. The study looked at the MDA in the testis tissues, as well as CAT, GPx, and GSH as antioxidants parameters, NF-& kappa;B and TNF-& alpha; as inflammatory markers, caspase-3 and Bcl-2 as apoptosis parameters, and PCNA as cell proliferation markers. In addition, testis tissues underwent histological evaluation. As a result, in rats exposed to only BPAF, sperm counts declined, testosterone levels reduced, oxidative stress, inflammation, and apoptosis increased, and cell proliferation decreased. Furthermore, severe disruptions in tissue architecture and decreased spermatogenesis were reported. In contrast, sperm parameters improved, testosterone levels increased, oxidative stress and inflammation decreased, and apoptosis was prevented in the carvacrol-treated group compared to the BPAF-only group. It was also found that spermatogenesis was maintained, and structural abnormalities in testicular tissue were mostly avoided with an increase in PCNA expression. According to the findings, despite BPAF-induced testicular and reproductive toxicity, carvacrol had therapeutic potential due to its anti-inflammatory, antioxidant, cell proliferation-increasing, and anti-apoptotic activities.
Comparison of different freezing techniques, extenders, and cryoprotectants on quality and fertility of cryopreserved Salmo trutta f. fario sperm
(Univ Estadual Maringa, Pro-Reitoria Pesquisa Pos-Graduacao, 2024) Bozkurt, Yusuf; Yavas, Ilker
There is still a lack of standardized methodology in the cryopreservation of Salmo trutta f. fario sperm. Thus, the present study was designed to compare current freezing protocols to improve and as well as to examine the post-thaw quality and fertilizing ability of cryopreserved sperm in Salmo trutta f. fario. Sperm samples were diluted at a 1:10 ratio in one of three extenders (Alsever's solution, glucose-based, and ionic-based) containing four types of cryoprotectant (DMSO, DMA, MeOH, glycerol) at 10% concentration and frozen in 0.1 mL pellets on surface of the dry ice (solid carbon dioxide, 79 degrees C) or in 0.25 mL straws 2 cm above of the liquid nitrogen (LN2) surface at a rate of similar to 30 degrees C for 10 min(-1) before storage in a mL cryotank (-196 degrees C). The frozen sperm cells in straws and pellets were thawed in a water bath at 25 degrees C for 30 s and at 20 degrees C for 6 s respectively. Fertilization was carried out using a ratio of 5x10(5) sperm/egg in both freezing methods. The glucose-based solution including glycerol produced the highest post-thaw progressive motility (62.5 +/- 1.24%), motility duration (57.2 +/- 0.46 s), and viability (56.4 +/- 1.57%) (p < 005) in the straw method. In breeding trials, similarly, sperm frozen-thawed with the glucose-based solution including glycerol produced the highest fertilization (54.2 +/- 0.36%) and hatching (30.6 +/- 0.28%) in the straw method. Fresh sperm used as control produced 82.6 +/- 0.45% and 78.4 +/- 1.27% fertilization and hatching respectively. It was concluded that sperm frozen in straws produced higher post-thaw sperm motility and fertility of eggs than those frozen in pellets. Also, the results suggest using of glycerol-supplemented glucose solution because of producing better results in both freezing techniques.
Cryopreservation of Brown Trout (Salmo trutta macrostigma) and Ornamental Koi Carp (Cyprinus carpio) Sperm
(Intech Europe, 2012) Bozkurt, Yusuf; Yavas, Ilker; Karaca, Fikret
[Abstract Not Available]
Cryopreservation of Nile Tilapia (Oreochromis niloticus) Sperm
(Intech Europe, 2016) Bozkurt, Yusuf; Yavas, Ilker
The main aim of this study is to determine the effect of the straw volume (0.25 vs. 0.5 mL) on Nile tilapia sperm quality after cryopreservation. Sperm was frozen according to conventional slow freezing procedure and diluted at ratio of 1: 3 with ionic extender containing 350 mM glucose and 30 mM Tris containing 10% dimethylacetamide. Diluted semen was equilibrated at 4 degrees C for 10 min and drawn into 0.25-mL or 0.5-mL plastic straws and sealed with polyvinyl alcohol. Samples were frozen 3 cm above of the liquid nitrogen surface and exposed to the liquid nitrogen vapor (approximate to-140 degrees C) for 10 min. After this, frozen sperm cells were kept into the liquid nitrogen container (-196 degrees C). The frozen sperm in different volume of straws were thawed in a water bath at 30 degrees C for 20 s (0.25-mL straws) or at 30 degrees C for 30 s (0.5-mL straws), respectively. Fertilization was conducted using 1 x 10(5) spermatozoa/egg ratio with each straw type. The findings of the present study indicated that cryopreservation of sperm in glucose-Tris-based extender using 0.5-mL straws improved post-thaw progressive motility, duration of progressive motility, and fertilization results (P<0.01). On the other hand, differences in term of post-thaw cell viability was not significant among the treatments (P>0.01). In conclusion, our results suggest that Nile tilapia sperm can be successfully cryopreserved in Tris-based extenders supplemented with glucose containing 10% dimethylacetamide in 0.5-mL straws.
Cryopreservation of scaly carp (Cyprinus carpio) sperm: effect of different cryoprotectant concentrations on post-thaw motility, fertilization and hatching success of embryos
(Springer, 2014) Yavas, Ilker; Bozkurt, Yusuf; Yildiz, Cengiz
The aim of the present study was to determine the effect of various cryoprotectants on post-thaw sperm quality and fertilizing capacity of cryopreserved scaly carp (Cyprinus carpio) semen. The present study focused on freezing of scaly carp sperm utilizing a practical and inexpensive protocol for aquaculture. Semen was diluted with Kurokura's extender composing 3.6 g/l NaCl, 10 g/l KCl, 0.22 g/l CaCl2, 0.08 g/l MgCl2 and 0.2 g/l NaHCO3. The extender contained three different cryoprotectants (DMSO, DMA and egg yolk) at ratios of 5, 10 and 15 %. Semen was placed into 0.25-ml straws and exposed to liquid nitrogen vapor (-120 A degrees C) using an insulated box with an adjustable tray for 10 min and then plunged into liquid nitrogen (-196 A degrees C) tank. The thawing process was performed in a water bath at 40 A degrees C for 10 s. The results indicated that type of cryoprotectants and their concentrations are rather effective in scaly carp sperm cryopreservation on post-thaw sperm quality, while they are very important in order to obtain high fertilization rates. The highest fertilization rate was determined as 96.4 +/- A 0.15 % with 15 % egg yolk, while the highest hatching rate was determined as 99.3 +/- A 0.80 with 15 % DMA. In conclusion, the applied cryopreservation method for scaly carp sperm is suitable to fertilize high amounts of eggs.
Cryoprotective Effect of Vitamin E Supplementation of Different Extenders on Quality and Fertilizing Ability of Frozen-Thawed Brown Trout Sperm
(Mary Ann Liebert, Inc, 2021) Bozkurt, Yusuf; Yavas, Ilker; Bucak, Mustafa Numan; Kiran, Tugba Raika; Gul, Aziz
Vitamin E is one of the most powerful antioxidants for prevention of cell damage resulting from cryopreservation, but its efficacy for cryopreserving brown trout sperm is still unclear. In this work, the protective effect of vitamin E on quality, fertilizing capacity, and DNA damage of brown trout (Salmo trutta macrostigma) sperm after cryopreservation was evaluated. Sperm samples were diluted at the ratio of 1:10 with three different extenders (E): (E-I): 300 mM glucose, 10% egg yolk; (E-II): 33.3 mM glucose, 5.1 mM NaCl, 0.5 mM NaHCO3,, 15% DMA; and (E-III): 61.6 mM NaCl, 134.2 mM KCl, 1.9 mM CaCl2, 0.8 mM MgCl2, 2.3 mM NaHCO3 in distilled water. Each extender was supplemented with 10% DMSO and different concentrations of vitamin E at 0.1, 0.5, and 1.0 mM. Spermatozoa frozen without vitamin E (0 mM, control) and fresh sperm were also used. After dilution, the sperm was aspirated into 0.25 mL straws, frozen 3 cm above the liquid nitrogen (LN2) surface, and plunged into the LN2. Cell motility, viability, fertilization, and eyeing were determined in post-thawed samples. DNA damage was determined by the comet assay after cryopreservation. Supplementation of 1 mM vitamin E to all extenders exhibited the best cryoprotective effect in terms of sperm motility, duration of motility, viability, fertility, and DNA integrity against cryopreservation damage, compared with 0.1, 0.5, and control group (0 mM) (p < 0.05). The highest post-thaw motility (62.4% +/- 0.36%), fertilization (48.2 +/- 0.84), and the lowest DNA damage (7.245%) were obtained with the extender-II including 1.0 mM vitamin E (p < 0.05). Consequently, vitamin E positively affected the motility parameters, fertility, and DNA integrity, and the results suggest the addition of extenders with vitamin E as an antioxidant for the cryopreservation of brown trout sperm.
Effect of cholesterol-loaded cyclodextrin on cryosurvival and fertility of cryopreserved carp (Cyprinus carpio) sperm
(Academic Press Inc Elsevier Science, 2015) Yildiz, Cengiz; Yavas, Ilker; Bozkurt, Yusuf; Aksoy, Melih
Addition of cholesterol-loaded cyclodextrin (CLC) to the diluents of mammalian semen increased stability and rigidity of phospholipid hydrocarbon chains of plasma membrane during sperm cryopreservation process. CLC has been tested successfully as ayoprotectant in various livestock sperm cryopreservation protocols but its efficacy for cryopreserving of fish sperm has not previously been tested. In the present study, different cholesterol loaded cyclodextrin concentrations were evaluated for the cryopreservation of carp (Cyprinus carpio) sperm. Sexually mature fish were induced to spermiation and ovulation with Ovopel. The extenders were prepared by using 300 mM glucose and 10% DMSO supplemented with different concentrations of CLC (0.5, 1.0, 1.5, 2.0, 2.5, and 3.0 mg per 120 x 10(6) spermatozoa) and without CLC (control). The pooled semen was diluted separately at a ratio of 1:3 (v/v) by using CLC extenders. Diluted semen placed into 0.25 ml straws were equilibrated at 4 degrees C for 15 min and frozen in liquid nitrogen vapor. Fertilization was conducted using a ratio of 1 x 105 spermatozoa/egg. Fresh sperm with no treatment showed the greatest sperm motility, duration of motility, viability, and fertilization results compared to the other tested cryopreserved and control groups (p < 0.05). Supplementation of 1.5 mg CLC to the extender showed the best cryoprotective effect for sperm motility, duration of motility, and viability against freezing damage in comparison to extenders containing 2.5 mg, 3.0 mg CLC, and control group (p < 0.05). Cryopreserved sperm containing 1.5 mg CLC provided greater result in term of fertilization success when compared to other extenders containing 0.5, 2.5, and 3.0 mg CLC or control (p <0.05). The amount of CLC effected post-thaw sperm quality and fertility as a dose-dependent manner. It is concluded that treatment of cholesterol-loaded cyclodextrin for carp sperm cryopreservation significantly improves cell cryosurvival and fertilization. (C) 2015 Elsevier Inc. All rights reserved.
Effect of different antioxidants on motility, viability and fertilizing capacity of cryopreserved scaly carp (Cyprinus carpio) semen
(Israeli Journal Of Aquaculture-Bamidgeh, 2014) Yavas, Ilker; Bozkurt, Yusuf; Yildiz, Cengiz
The present study investigated effect of different antioxidants such as taurine (25 mM, 50 mM, 75 mM and 100 mM), bovine serum albumine (BSA) (1%, 2.5%, 5%, 7.5%) and vitamin C (ascorbic acid) (2.8 mM, 5.6 mM, 8.4 mM and 11.2mM) on motility and fertilizing ability of cryopreserved scaly carp (Cyprinus carpio) semen. The results demonstrated that addition of taurine at doses of 50 mM and 75 mM and also BSA at doses of 2.5% and 5% improved fertilization ability of cryopreserved scaly carp semen. The highest post-thaw motility (42.4 +/- 1.5%), viability (67.8 +/- 2.6%), fertilization (59.3 +/- 0.5%) and also hatching rates (68.2 +/- 3.4%) were determined by supplementation of 75 mM taurine (p<0.05). These results suggest that supplementation of ionic extender with taurine is suitable for scaly carp semen cryopreservation.
Effect of different avian egg yolk types on fertilization ability of cryopreserved common carp (Cyprinus carpio) spermatozoa
(Springer, 2014) Bozkurt, Yusuf; Yavas, Ilker; Yildiz, Cengiz
In spite of the fact that egg yolk from different avian species has successfully been used as an additive for the cryopreservation of sperm in mammalian species, its efficacy for cryopreserving fish sperm has not previously been tested comparatively. Therefore, the present study was carried out to determine the effect of egg yolks from different avian species, namely domestic chicken (Gallus gallus domesticus), turkey (Meleagris gallopavo) and quail (Coturnix coturnix), on post-thaw motility and fertilization ability of cryopreserved common carp spermatozoa. Egg yolks from chicken, turkey and quail were analysed for moisture, total fat, protein, cholesterol and phospholipid profile. Total fat and cholesterol contents of the turkey egg yolk were higher than chicken and quail egg yolks (p < 0.05). Semen was frozen according to conventional slow freezing procedure. The extender contained 350 mM glucose, 30 mM Tris and 5 % glycerol supplemented with different ratios of avian egg yolk (10, 15 and 20 %). Semen was equilibrated at 4 A degrees C for 15 min and placed into 0.25-ml straws and frozen in liquid nitrogen vapour (for 10 min at -120 A degrees C) and finally stored in liquid nitrogen (-196 A degrees C) tank. The frozen spermatozoa were thawed in a water bath at 35 A degrees C for 30 s. Fertilization was conducted using a ratio of 1 x 10(5) spermatozoa/egg. Cryopreservation experiments resulted in higher post-thaw motility and fertilization rates. Mean post-thaw motility of cryopreserved spermatozoa was between 45 and 80 %, and fertilization rates, expressed as the percentage of eyed embryos, ranged from 70 to 95 %. In conclusion, the present study showed that turkey and quail egg yolks are suitable alternatives to the chicken egg yolk for the cryopreservation of common carp spermatozoa.
EFFECT OF DIFFERENT CRYOPROTECTANTS (GLYCEROL, METHANOL AND DIMETHYL SULFOXIDE) ON POST-THAW QUALITY, VIABILITY, FERTILIZATION ABILITY AND DNA DAMAGE OF CRYOPRESERVED NILE TILAPIA (Oreochromis niloticus) SPERMATOZOA
(Cryo Letters, 2019) Bozkurt, Yusuf; Yavas, Ilker; Bucak, Mustafa Numan; Yeni, Deniz
BACKGROUND: Cryopreservation of sperm from different fish species requires different protocols. Therefore, it is necessary to perform studies to establish reliable procedures for each species. OBJECTIVE: Experiments were designed to analyse the effect of different types of cryoprotectants on post-thaw motility, viability and fertility as well as cryoresistance of cryopreserved Nile tilapia (Oreochromis niloticus) sperm. MATERIALS AND METHODS: Sperm samples were diluted with an ionic extender containing glycerol (Gly), methanol (MeOH) and dimethyl sulfoxide (DMSO) at ratios of 5, 10 and 15 % respectively. Diluted samples were aspirated into 0.25 ml French straws and frozen 3 cm above the surface of liquid nitrogen (LN) in a styrofoam box and stored in a LN tank. DNA damage was evaluated with the comet assay technique following cryopreservation. RESULTS: Supplementation of extender with 10% glycerol gave the highest motility rate compared with the other cryoprotectant groups (P<0.05). Differences in terms of post-thaw motility duration, cell viability and fertilization rates were not significant among treatments (P>0.05). Although Gly gave the best score (5.0 +/- 0.1%, P>0.05) at the concentration of 10%, 5% Me2SO caused significant DNA damage (15.0 +/- 1.0%, P<0.05) with the comet test. CONCLUSION: Gly or MeOH are more suitable cryoprotectants than DMSO for the cryopreservation of Nile tilapia sperm.
Effect of different plant extracts on microbial population in the frozen African Catfish (Clarias gariepinus) semen
(C M B Assoc, 2023) Secer, Faik Sertel; Yavas, Ilker; Cantekin, Zafer; Bozkurt, Yusuf; Yavas, Tugba Korkmaz; Atanasoff, Alexander
Fish sperm cryopreservation has been attempted on roughly freshwater and marine species since 1953. This study sought to assess the potential of various plant extracts to function as natural antimicrobial agents in the frozen semen of African catfish (Clarias gariepinus). Diluted sperm was packaged in 0.25ml straws and left for 10min equilibration at 4 degrees C. Following equilibration, the straws were exposed to liquid nitrogen vapor for 10 min and plunged into the liquid nitrogen (-196 degrees C) and then thawed in a water bath at 35 degrees C for 20s. Sperm samples were put into sterile 1.5 ml tubes immediately after thawing and the microbial count was detected with classical microbiological culture method. In the results of microbiological analyses, these tree plant extracts especially Echinacea purpurea were found highly effective for decreasing bacterial contamination levels of African catfish (C. gariepinus) semen. These plant extracts may have the potential for antibacterial effect, and they can be useful for the dilution of semen.
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.
EFFECT OF DIFFERENT THAWING RATES ON MOTILITY AND FERTILIZING CAPACITY OF CRYOPRESERVED GRASS CARP (CTENOPHARYNGODON IDELLA) SPERM
(Taylor & Francis Ltd, 2011) Yavas, Ilker; Bozkurt, Yusuf
Cryopreservation of spermatozoa has been well developed in many fish species for resource conservation and aquaculture practices. There are limited data on the effect of cryopreservation on grass carp (Ctenopharyngodon idella) spermatozoa. This research was carried out to investigate the effect of different thawing procedures on motility and fertilizing capacity of frozen! thawed grass carp semen as preliminary data to design future cryopreservation experiments. Semen was collected by abdominal stripping from adult male grass carp. Collected semen was diluted with extender containing 350 mM glucose, 30 mM Tris and 5% glycerole (pH 8.0). Diluted samples were packaged in 0.25 ml straws and left to equilibrate for 30 min at4 degrees C. Following equilibration, the straws were exposed to liquid nitrogen vapour for 10 min and plunged into the liquid nitrogen (-196 degrees C). For thawing, the straws were removed from liquid nitrogen and immersed in 30, 35 and 40 C water for 10, 20 and 30 seconds. In fertilization experiments spermatozoa:egg ratio was 1x10(5):1. The highest mean motility (83.4 +/- 2.1) (p<0.05) and fertilization rate (85.6 +/- 2.8) (P<0.05) was determined from semen thawed at 35 C for 30 sec. The results indicate that thawing rates significantly influenced motility and fertilizing ability of cryopreserved grass carp semen.
Effect of exogenous Melatonin administration on Spermatogenesis in chronic unpredictable stress rat model
(Univ Zulia, Facultad Ciencias Veterinarias, 2023) Gokcek, Ishak; Aydin, Leyla; Cellat, Mustafa; Yavas, Ilker; Kutlu, Tuncer
This study investigated the hormonal, inflammatory, oxidant-antioxidant, and histopathological effects of exogenous Melatonin administration on Spermatogenesis in rats' chronic unpredictable stress model (CUSM). In the study, stress caused a decrease in follicle stimulating-hormone (FSH), luteinizing hormone (LH), Testosterone, Melatonin, Glutathione (GSH), Glutathione peroxidase (GSH-Px), catalase, interleukin 10 (IL-10) levels and motility, and an increase in Corticosterone, nuclear factor kappa beta (NF-kB), tumor necrosis factor-alpha (TNF-alpha), interleukin 1 beta (IL-1 beta), interleukin 6 (IL-6), abnormal sperm, dead/live sperm ratio and exogenous Melatonin reduced inflammatory cytokines and oxidative stress and improved spermatological parameters (P<0.05). Melatonin also partially corrected stress-induced changes in testicular morphology. As a result, using Melatonin in rats with CUSM may be effective in improving spermatological parameters through anti-inflammatory and antioxidant mechanisms.
Effect of Exogenous Melatonin Chronic Unpredictable Stress Model on Spermatogenesis in Rats
(Wiley, 2022) Gokcek, Ishak; Aydin, Leyla; Cellat, Mustafa; Yavas, Ilker
[Abstract Not Available]
Effect of Extender Compositions, Glycerol Levels, and Thawing Rates on Motility and Fertility of Cryopreserved Wild African Catfish (Clarias gariepinus) Sperm
(Israeli Journal Of Aquaculture-Bamidgeh, 2017) Bozkurt, Yusuf; Yavas, Ilker
The aim of this study was to determine the effect of different extenders, glycerol levels, and thawing rates on post-thaw sperm motility and fertilization ability of cryopreserved African catfish (Clarias gariepinus) sperm. Having determined the main spermatological properties (volume, motility, motility duration, spermatozoa concentration and pH), the pooled ejaculates were diluted with 3 different extenders containing different glycerol levels (5, 10 and 15%) respectively. Dilution ratio was 1:10 and the diluted sperm was packaged in 0.25 ml straws and left for 10 min equilibration at 4 degrees C. Following equilibration, the straws were exposed to liquid nitrogen vapor for 10 min and plunged into liquid nitrogen (-196 degrees C), and then exposed to different thawing rates (30 degrees C/20s and 40 degrees C/20s) to determine sperm motility and post-thaw motility duration. The highest post-thaw sperm motility, motility duration, and fertilization rate was 85%, 81s, and 95% respectively when sperm was frozen with the extender (ACSE 3) containing 15% glycerol (p < 0.05). The protocol reported in this study can be successfully used for cryopreservation of African catfish sperm.
EFFECT OF EXTENDER SUPPLEMENTED WITH BORON ON POST-THAW MOTILITY, VIABILITY, DNA DAMAGE AND FERTILIZATION ABILITY OF CRYOPRESERVED BROWN TROUT (Salmo trutta macrostigma) SPERMATOZOA
(Cryo Letters, 2019) Bozkurt, Yusuf; Yavas, Ilker; Gul, Aziz; Bucak, Mustafa Numan; Yeni, Deniz; Avdatek, Fatih
BACKGROUND: Boron has been considered as an essential nutrient for decreasing lipid peroxidation and improving antioxidant mechanism in different animal species. On the other hand, its effect on quality or DNA damage following cryopreservation process in fish sperm is still unclear. OBJECTIVE: Experiments were designed to analyse the effect of an ionic based extender supplemented with boron on post-thawed motility, viability, fertility and DNA integrity of cryopreserved brown trout (Salmo trutta macrostigma) sperm. MATERIALS AND METHODS: Sperm samples were cryopreserved with the ionic extender containing different boron concentrations (0.1, 0.2, 0.3 and 0.4 mM) using a controlled freezer at two different freezing rates (FR-I: 10 degrees C min(-1) from +4 degrees C to -40 degrees C and FR-II: 15 degrees C min(-1) from +4 degrees C to -40 degrees C). Sperm motility, viability, fertilization, eyeing and DNA fragmentations were determined in post-thawed samples. RESULTS: Freezing rate-I provided significantly higher fertilization and eyeing rates compared to freezing rate-II (p<0.05). Higher post-thaw motility (62.8 +/- 1.4%) and fertilization (75.2 +/- 0.9%) rates were obtained with the 0.4 mM boron concentration at freezing rate-I. CONCLUSION: Supplementation of the extender with boron increased fertilization and eyeing rates and also decreased DNA damages at both freezing rates.
Effect of Extender Supplemented with Different Sugar Types on Post-thaw Motility, Viability and Fertilizing Ability of Cryopreserved Common Carp (Cyprinus carpio) Spermatozoa
(Israeli Journal Of Aquaculture-Bamidgeh, 2016) Bozkurt, Yusuf; Yavas, Ilker; Yildiz, Cengiz
The influence of various sugar types supplemented to the extender on post-thaw motility, viability, and fertilizing capacity of cryopreserved common carp (Cyprinus carpio) semen were investigated. The results indicated that types of sugar significantly influenced motility, motility duration, and viability rates (P<0.05). Glucose, maltose, sucrose, and trehalose provided higher motility compared to the sugar-free control in post-thaw samples. Trehalose provided highest progressive motility duration, and higher viable sperm rates were obtained with all sugar types except xylose. Xylose exhibited the lowest post-thaw progressive motility duration (35.2 +/- 1.4s). The mean highest fertilization (78.2 +/- 1.4%) and eyed egg rates (94.3 +/- 1.5%) were determined using trehalose in the extender, and differences between the treatments in the fertilization and eyeing rates were significant (P<0.05). Finally, the present study showed that sugars, especially maltose and trehalose (disaccharide), improved post-thaw spermatozoa motility and fertility in common carp semen.
Effect of Glycerol on Fertility of Cryopreserved Grass Carp (Ctenopharyngodon idella) Sperm
(Israeli Journal Of Aquaculture-Bamidgeh, 2011) Bozkurt, Yusuf; Yavas, Ilker; Ogretmen, Fatih; Sivasligil, Bugra; Karaca, Fikret
This research investigated the effect of adding glycerol to different ionic extenders on motility and fertilizing ability of frozen-thawed sperm of grass carp (Ctenopharyngodon idella) under hatchery conditions. Semen was collected by abdominal stripping from adult males and diluted to a ratio of 1:3 in Kurokura I or Kurokura II extender containing 10%, 15%, or 20% glycerol. Diluted semen was packaged in 0.5-ml straws and left to equilibrate for 30 min at 4 degrees C. Following equilibration, the straws were exposed to liquid nitrogen vapor for 10 min and plunged into liquid nitrogen (-196 degrees C). Straws were thawed in a water bath at 30 degrees C for 10 s to determine the post-thaw motility and movement duration. The spermatozoa: egg ratio in fertilization experiments was 1 x 10(5):1. The highest mean fertilization rate (95.63 +/- 0.52; p>0.05) was obtained with semen frozen in Kurokura I extender containing 10% glycerol. Results indicate that grass carp sperm can be successfully cryopreserved with ionic extenders containing glycerol at 10-20%.

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