Esterified glucomannan improves aflatoxin-induced damage of sperm parameters during liquid storage of ram semen at 5°C

The aim of the present work was to study the effects of aflatoxin (AF) on sperm parameters in rams, and to determine the protective efficiency of esterified glucomannan (EG) co-administered with AF up to 96. h of the liquid storage of ram semen at 5. °C. Thirty-two Merino rams (12-14. months old) were used. The animals were examined for their general health status. To ensure their adaptation to the environment and the new feeding regimen, a 15-day acclimatization programme was applied to the animals, prior to the start of the study. Experimental feeding was continued for ninety-two days. The experimental design consisted of four dietary treatments. The control group (C) was fed with commercial feed. The AF group was fed with commercial feed plus 250. μg/day of total AF. The EG group received commercial feed plus 2. g/day of EG. The AF + EG group was given commercial feed plus 250. μg/day of total AF and 2. g/day of EG. In the study, ejaculates were obtained from rams twice a week for 12. weeks, using an electro-ejaculator. After collected, the ejaculates were diluted with a skimmed milk extender, and stored at 5. °C. Sperm motility and rates of abnormal and nonviable spermatozoa were determined for the different treatment groups at 5. °C at 0, 24, 48, 72 and 96. h of liquid storage.During the first two weeks of the trial, the groups did not statistically differ from each other for sperm motility or rates of abnormal and nonviable spermatozoa at 0, 24, 48 and 96. h of storage. As from the third week, the short-term storage of semen produced statistically significant differences between the AF group and the other treatment groups for sperm parameters (p<. 0.05). The administration of aflatoxin was observed to have reduced sperm motility and to have increased the rates of abnormal and nonviable spermatozoa in comparison to the control group (p<. 0.05), while EG co-administered with AF was determined to have ameliorated the adverse effects of AF on sperm parameters, and this ameliorative effect continued throughout the short-term storage of semen. On the other hand, aflatoxin administration resulted in the deterioration of the sperm parameters in the following weeks, and the combined administration of EG + AF reversed this adverse effect, thus, bringing the sperm parameters closer to the values of the control group. This study demonstrated that, in rams, AF adversely affected sperm, biochemical and testis parameters, and that the combined administration of EG and AF reversibly improved these adverse effects. © 2014 Elsevier Inc

Relationship of blood and seminal plasma ceruloplasmin, copper, iron and cadmium concentrations with sperm quality in Merino rams

The aim of the current study was to investigate the concentrations of ceruloplasmin, copper, iron, zinc and cadmium concentrations in blood serum and seminal plasma obtained from Merino rams. In addition, their relationship with sperm parameters, fertility rate and litter size were also studied. Blood and ejaculate samples (6 replicates) were taken in October from 19 Merino rams, aged between 18 and 24 months. Ceruloplasmin, copper, iron, zinc and cadmium in blood serum and seminal plasma were determined. Sperm parameters including volume, mass motility, motility, concentration, Hos-test, viability, abnormal sperm and acrosome abnormality in semen, fertility rate and litter size were also evaluated. Highly positive correlation was found between blood ceruloplasmin and blood copper concentrations (r = 0.812, p 0.001), whereas negative correlation were determined between these parameters in seminal plasma (r = -0.195, p 0.05). Seminal plasma copper concentration was positively correlated with seminal plasma cadmium (r = 0.206, p 0.05) and seminal plasma iron (r = 0,305, p 0.01) concentrations. Negative correlation was determined between blood ceruloplasmin level and acrosomal defect (r = -0.443, p 0.05). Seminal plasma ceruloplasmin level was positively correlated with volume (r = 0.255, p 0.01) and negatively correlated with abnormal sperm (r = -0.186, p = 0.058) and acrosome abnormality (r = -0.213, p 0.05). Seminal plasma iron concentration was positively correlated with other abnormality (r = 0.257, p 0.01). Seminal plasma cadmium concentration was positively correlated with sperm abnormality (r = 0.207, p = 0.052) and other abnormality (r = 0.262, p 0.05) and negatively correlated with fertility rate (r = -0.449, p = 0.054). Blood cadmium concentration was negatively correlated with litter size (r = -0.579, p 0.01). In conclusion, blood and seminal plasma ceruloplasmin may be suggested to have positive influence regardless of copper with its antioxidant property whereas iron and cadmium have negative influence on sperm parameters and fertility in Merino rams. © 2015 Elsevier B.V

The synergistic effect of trehalose and low concentrations of cryoprotectants can improve post-thaw ram sperm parameters

The objective of this study was to compare the effects of different concentrations of two different cryoprotectants (glycerol, G and ethylene glycol, EG) and trehalose (T), added to the semen extender, on post-thaw ram sperm parameters. Ejaculates, collected from 6 Merino rams, were pooled and evaluated at 37 °C. The pooled samples were divided into six equal aliquots, and diluted in Tris-based extenders containing 5% G, 3% G + 60 Mm T, 1.5% G + 100 Mm T, 5% EG, 3% EG + 60 mM T, and 1.5% EG + 100 Mm T. Subsequently, the samples were cooled to 5 °C, frozen in 0.25-ml French straws, and stored in liquid nitrogen (LN2). Frozen samples were thawed individually, at 37 °C for 25 s in a water bath, for evaluation. Sperm motility was assessed using a phase-contrast microscope with a warm stage maintained at 37 °C. Acrosome integrity (FITC/PNA-PI), sperm viability (SYBR-14/PI), mitochondrial activity (JC-1/PI), DNA damage (COMET assay) and DNA fragmentation (TUNEL test) were determined. The group of samples diluted in an extender containing 5% of glycerol (Group 5% G) displayed higher percentages of subjective motility, viability and mitochondrial activity of sperm, compared to the other groups (P ;lt; 0.05). On the other hand, Group 3% G + 60 mM T yielded the second-best results for subjective motility, viability and mitochondrial activity of sperm, when compared to the other groups. The post-thaw sperm parameters of Group 3% G + 60 Mm T did not show any statistically significant difference from those of Group 5% G. There were no statistically significant differences between the groups for acrosome integrity (P ;gt; 0.05). The results of the COMET assay showed that the use of low concentrations of cryoprotectants in combination with trehalose decreased sperm DNA damage. Accordingly, Group 1.5% G + 100 mM T and Group 3% EG + 60 mM T benefited from a significantly stronger cryoprotective effect on DNA integrity, in comparison to Group 5% G (P ;lt; 0.05). According to the results of the TUNEL test, the combined use of low concentrations of cryoprotectants with trehalose decreased sperm DNA damage, when compared to the use of 5% glycerol, but this difference was statistically insignificant (P ;gt; 0.05). In conclusion, G and EG concentrations can be reduced by adding various amounts of T (60 mM, 100 mM) to the semen extender. The addition of 5% of glycerol and 3% G + 60 mM T to the semen extender did not yield statistically different post-thaw sperm parameters, when compared for protection against cryoinjury. Post-thaw sperm parameters can be improved by the supplementation of the semen extender with 3% G + 60 mM T. Thus, we recommend the use of freezing extenders containing low cryoprotectant concentrations (3% G) combined with trehalose to avoid the high level of toxic and osmotic damage caused by 5% G. © 2020 Elsevier Inc

Decreasing glycerol content by co-supplementation of trehalose and taxifolin hydrate in ram semen extender: Microscopic, oxidative stress, and gene expression analyses

This study aimed to evaluate the comparative effects of taxifolin hydrate and trehalose on the quality of frozen-thawed ram spermatozoa for the first time. Ejaculates collected from six mature rams were pooled, and divided to eight equal aliquots to extend them with different concentrations of glycerol (%5 and %3), taxifolin hydrate (10, 100, and 500 µM), and trehalose (60 mM) as eight groups (G5T0, G5T10, G5T100, G5T500, G3T0, G3T10, G3T100, and G3T500). After freeze-thawing process of cryopreservation, microscopic and oxidative stress parameters, and gene expression levels were investigated for understanding of possible impacts of taxifolin hydrate and trehalose. The study showed that G3T10 resulted in the highest post-thawed viability and mitochondrial activity. Moreover, all extenders with taxifolin hydrate reduced DNA fragmentation in comparison to G5T0, but DNA damage was prevented at the highest rate in presence of G5T10. The level of LPO significantly decreased in the groups G5T500 and G3T100, and the expression levels of NQO1, GCLC, and GSTP1 genes significantly increased in the groups G5T100, G5T500, G3T10, and G3T100 compared to the group G5T0. Finally, co-supplementation of tris-based extender having 3% glycerol with 60 mM trehalose and 10 µM taxifolin hydrate in cryopreservation extender may be recommended to improve the quality of post-thawed ram spermatozoa. However, further in vivo and in vitro studies are suggested to evaluate fertility rates of frozen-thawed ram spermatozoa co-supplemented with trehalose and taxifolin hydrate. © 2020 Elsevier Inc

Raffinose and hypotaurine improve the post-thawed Merino ram sperm parameters

The aim of this study was to determine the effects of raffinose and hypotaurine on sperm parameters after the freeze-thawing of Merino ram sperm. Totally 40 ejaculates of five Merino ram were used in the study. Semen samples, which were diluted with a Tris-based extender containing 10. mM raffinose, 5. mM hypotaurine, 5. mM raffinose +2.5. mM hypotaurine (H + R) and no antioxidant (control), were cooled to 5 °C and frozen in 0.25. ml French straws and stored in liquid nitrogen. Frozen straws were then thawed individually at 37 °C for 25. s in a water bath for evaluation. The addition of raffinose led to higher percentages of subjective and CASA motilities (47.5 ± 12.2%, 46.3 ± 13.6%) compared to controls (38.8 ± 13.8%, 30.5 ± 11.7%, P 0.05). For the CASA progressive motility, 5. mM raffinose (20.12 ± 8.82%) had increasing effect in comparison to control (10 ± 7.94%, P 0.05) following the freeze-thawing process. Raffinose and hypotaurine led to higher viability (40.8 ± 4.68%, 40.8 ± 4.7%), high sperm mitochondrial activity (29.5 ± 5.4%, 27.3 ± 4.9%) and acrosome integrity (50.8 ± 8.1, 50.7 ± 4.4) percentages, compared to control groups (31.5 ± 3.5%, 9.5 ± 8.2%, 42.8 ± 7.3%, P 0.05). H + R group only led to high sperm mitochondrial activity when compared to control group. In the comet test, raffinose and hypotaurine resulted in lower sperm with damaged DNA (6.2% and 3.9%) than that of control (9.1%), reducing the DNA damage. For TUNEL assay, The TUNEL-positive cell was distinguished by distinct nuclear staining. Raffinose and H + R groups resulted in lower sperm with TUNEL-positive cell (1.5 ± 1.2% and 2.1 ± 0.9%) than that of control (4.9 ± 2.5%) (P 0.05). In conclusion, findings of this study showed that raffinose and hypotaurine supplementation in semen extenders provided a better protection of sperm parameters against cryopreservation injury, in comparison to the control groups. © 2013

Influence of lycopene and cysteamine on sperm and oxidative stress parameters during liquid storage of ram semen at 5 °C

Ejaculates were collected from six Merino rams with the aid of an artificial vagina twice a week. The ejaculates containing spermatozoa with ;gt;80% forward progressive motility and concentrations higher than 2 × 109 spermatozoa/ml were pooled. The present study included two experiments. In experiment 1, each pooled ejaculate was divided into four equal aliquots and diluted (37 °C) with the Tris based extender, containing 0 (control), 0.5, 1 and 2 mM lycopene, at a final concentration of approximately 400 × 106 sperms/ml (single step dilution), In experiment 2, cysteamine at concentrations of 0 (control), 0.5, 1 and 2 mM, was used as an additive in the extender, and the procedure explained above was applied for the division of aliquots and the dilution of semen. Diluted semen samples were kept in glass tubes and cooled from 37° to 5 °C in a cold cabinet, and maintained at 5 °C. Sperm and oxidative stress parameters were evaluated after 0, 24, 48 and 72 h of storage at 5 °C. The extender supplemented with 0.5 mM lycopene resulted in higher mitochondrial activity rate (p ;lt; 0.05) in comparison to the control group at 72 h of storage. Lycopene at 0.5 mM dose led to higher sperm motility rate (p ;lt; 0.05) when compared to 2 mM lycopene group at 72 h of liquid storage. As regards oxidative stress parameters, only 2 mM lycopene increased total glutathione levels (p ;lt; 0.05) at 0 h of storage. The extender supplemented with 1 mM cysteamine gave higher motility (p ;lt; 0.05) at 48 h compared to control. As regards oxidative stress parameters, 1 and 2 mM cysteamine at 48 h and 1 mM cysteamine at 72 h increased total glutathione levels (p ;lt; 0.05) compared to control groups. Cysteamine at 1 and 2 mM doses decreased lipid peroxidation (p ;lt; 0.05) at 0 h of liquid storage compared to control. Our data suggest that lycopene at 0.5 and 2 mM and cysteamine at 1 and 2 mM doses can be added to Tris based extender for improving the ram sperm motility, viability, mitochondrial activity and oxidative stress parameters during the liquid storage. © 2016 Elsevier B.V

Relationship of blood and seminal plasma ceruloplasmin, copper, iron and cadmium concentrations with sperm quality in Merino rams

The aim of the current study was to investigate the concentrations of ceruloplasmin, copper, iron, zinc and cadmium concentrations in blood serum and seminal plasma obtained from Merino rams. In addition, their relationship with sperm parameters, fertility rate and litter size were also studied. Blood and ejaculate samples (6 replicates) were taken in October from 19 Merino rams, aged between 18 and 24 months. Ceruloplasmin, copper, iron, zinc and cadmium in blood serum and seminal plasma were determined. Sperm parameters including volume, mass motility, motility, concentration, Hos-test, viability, abnormal sperm and acrosome abnormality in semen, fertility rate and litter size were also evaluated. Highly positive correlation was found between blood ceruloplasmin and blood copper concentrations (r = 0.812, p < 0.001), whereas negative correlation were determined between these parameters in seminal plasma (r = -0.195, p < 0.05). Seminal plasma copper concentration was positively correlated with seminal plasma cadmium (r = 0.206, p < 0.05) and seminal plasma iron (r = 0,305, p < 0.01) concentrations. Negative correlation was determined between blood ceruloplasmin level and acrosomal defect (r = -0.443, p < 0.05). Seminal plasma ceruloplasmin level was positively correlated with volume (r = 0.255, p < 0.01) and negatively correlated with abnormal sperm (r = -0.186, p = 0.058) and acrosome abnormality (r = -0.213, p < 0.05). Seminal plasma iron concentration was positively correlated with other abnormality (r = 0.257, p < 0.01). Seminal plasma cadmium concentration was positively correlated with sperm abnormality (r = 0.207, p = 0.052) and other abnormality (r = 0.262, p < 0.05) and negatively correlated with fertility rate (r = -0.449, p = 0.054). Blood cadmium concentration was negatively correlated with litter size (r = -0.579, p < 0.01). In conclusion, blood and seminal plasma ceruloplasmin may be suggested to have positive influence regardless of copper with its antioxidant property whereas iron and cadmium have negative influence on sperm parameters and fertility in Merino rams. © 2015 Elsevier B.V

Lipid mixtures (from a liposome kit) and melatonin improve post-thawed Angora goat sperm parameters

Semen freezing and storing has been widely used in reproductive biotechnology, being applied to certain males of livestock breeds or animal species with economic value such as the Angora goat. The development of a semen extender with the cryoprotective agents can prevent the deterioration of sperm parameters after thawing. This study aimed to investigate lipid mixtures (from a liposome kit, Lps) and melatonin (Mel) at different doses to prevent the deterioration of sperm parameters and to provide the cryoprotective effects on sperm DNA. The Angora goat ejaculates were collected and pooled. They were divided into seven equal volumes, and each of them was diluted with the extenders of the experimental groups with additives (Lps 321.99 μg/mL, Lps 841.33 μg/mL, Mel 0.25 mM, Mel 1 mM, Lps 321.99 μg/mL + Mel 1 mM, Lps 841.33 μg/mL + Mel 0.25 mM) and no additives (control group). After the freeze-thawing process, motility, viability, acrosome integrity, DNA double-strand breaks, and abnormal DNA integrity were assessed for different extender groups. It was determined that the use of Lps alone at low dose or the combination of Lps and Mel had significant cryoprotective effects on motility, viability, acrosome integrity, and DNA damage in Angora goat sperm. This study will help us to understand the effects of Lps and Mel used alone or in combination at different doses and which doses give the optimum spermatological parameter rates following the freeze-thawing process, and hence it will shed light on further studies. © 2024 Society for CryobiologySelçuk Üniversitesi, SÜ: 21212007; Selçuk Üniversitesi, S