Supplementation of Baicalin (BC) in extender improves structural and functional characteristics, total antioxidant capacity, and in vivo fertility of ram semen.

The effects of adding mixed chicken and Japanese quail egg yolks (EYs) to the cryodiluent on the quality of ram semen before freezing and post-thawing were evaluated. Additionally, the composition of chicken and quail egg EYs and their mixture were analyzed for results explanation. The semen was collected from rams (n = 5) and extended with cryodiluent containing the EY of chicken, quail or their mixture (1:1). The extended semen was chilled slowly to 5 °C within 2 h and equilibrated for 2 h, before frozen on the liquid nitrogen vapor and cryopreserved at −196 °C. The straws were evaluated before freezing and post-thawing for sperm motility, vitality and abnormality besides plasma-membrane and DNA integrities. The moisture, ash, protein, and fatty acid (FA) contents of chicken EY, quail EY and their mixture were analyzed. Sperm vitality, plasma membrane integrity and DNA integrity before freezing were significantly (P < 0.05) higher in quail EY than chicken EY and mixed EYs cryodiluent. The chicken EY extender significantly improved the vitality, plasma membrane and DNA integrities of post-thawed ram semen in comparison with quail EY or mixed EYs extenders. While, the post-thawing sperm abnormalities was lower (P ≤ 0.05) in quail EY than chicken EY and mixed EYs cryodiluent. The post-thawing sperm motion kinetics parameters were higher in quail EY than chicken EY and mixed EYs cryodiluent. The highest percentages of moisture, ash, saturated fatty acids (SFAs) and monounsaturated fatty acids (MUFAs) were detected in quail EY had. While, the highest percentages of fat, protein and polyunsaturated fatty acids (PUFAs) were detected in chicken EY. In conclusion, using of chicken EY can improve total motility, vitality, plasma membrane integrity and DNA integrity of cryopreserved ram semen. While, using of quail EY can improve sperm abnormalities and kinetic motion parameters of cryopreserved ram semen. Mixing chicken and quail EYs added no value for post-thawing ram semen parameters.

Effects of astaxanthin supplementation on the sperm quality and antioxidant capacity of ram semen during liquid storage

"The study was carried out to evaluate some microscopic parameters (mobility, concentration, viability), to assess the metabolic intensity of spermatozoa (Redox test), and sperm resistance test related with ram fertility and the quality of ram semen in different age groups. The study was conducted in a farm located in Cluj County, on 34 rams of Turcana Alba breed, grouped according to age into 4 batches. Weekly an ejaculate and the mentioned variables were measured. Semen samples were collected from each animal using the artificial vagina (AV). For sperm mobility, the best values were observed for 3 years old rams (X ± S = 88.4 ± 3.02). Variations in sperm viability showed some changes, but for all age groups were obtained values above those indicated in the literature. Assessment of sperm concentration revealed that rams in B6 (X ± S = 2.75 ±0.31) and B5 (X ± S = 2.7 ± 0.38) had the best values. Higher metabolic intensity rate in B6, B5, B3 groups was correlated with higher values of concentration and mobility in these age groups. Thus, the best values regarding sperm resistance were recorded for the rams aged 6 and 5 years, in which the average values were equal to 7022.22. With increase in age, ram showed increase percentages of motility and viability of sperm in all studied batches."

The present study determined the effects of conventional, and controlled freezing method adopting three freezing rates, viz. 20°C, 40°C and 60°C/min on quality of sperm (motility, viability and plasma membrane integrity), DNA integrity and plasma membrane protein profile of cryopreserved boar semen. Sixty sperm-rich fractions of ejaculates from six boars were utilized for freezing of semen with different freezing methods in lactose-egg yolk glycerol extender. Semen samples were evaluated for sperm motility, viability (Propidium Iodide assay), functional integrity of plasma membrane (HOST), DNA integrity (Acridine Orange stain) and plasma membrane protein profile (SDSPAGE) after equilibration and after freezing. The results revealed that the post thaw sperm motility, sperm viability, and plasma membrane integrity (HOST-reacted) were significantly higher in all the three controlled freezing methods (20°C, 40°C and 60°C/min) as compared to that in conventional method. In addition, the number of sperm plasma membrane protein loss was less in controlled freezing methods as compared to that in conventional freezing. However, the post thaw sperm DNA integrity did not influence by difference in freezing methods. No significant difference on the post thaw sperm characteristics was recorded among the three controlled freezing rates. All the sperm parameters assessed declined significantly after freezing as compared to that after equilibration irrespective of freezing method employed. In conclusion, controlled freezing methods conferred better post thaw sperm quality as compared to conventional method, and thus the freezing rates of either 20, 40 or 60°C/min could provide better freezability of boar semen.

The objective of this research was to investigate the effect of astaxanthin supplementations of semen extender on the quality of Hu ram semen after up to five days of preservation at 4 °C. Semen samples were collected from five healthy Hu rams using an artificial vagina during breeding season (April to August 2023) and diluted with a basic extender supplemented with control (0), 1 µM, 2 µM, 3.5 µM, or 4.5 µM of AXT. Overall, 170 semen ejaculate samples (34 repetitions) from five healthy Hu rams were used in our research study. The results revealed that the addition of AXT (3.5 µM) significantly (p ≤ 0.05) increased the sperm kinematic indexes (T.M%, P.M%, MAD%, STR%, and LIN %), sperm viability, plasma membrane integrity, acrosome integrity, total antioxidant content (T-AOC), and mitochondrial membrane potential (MMP) of the Hu rams spermatozoa after up to five days of preservation at 4 °C. Contrary to that, the addition of the best concentration of AXT (3.5 µM) to the semen extender significantly (p ≤ 0.05) reduced the reactive oxygen species (ROS) and malondialdehyde (MDA) concentration of Hu ram semen. In conclusion, the results of the current study indicate that the addition of a semen extender with AXT improves the quality of Hu ram spermatozoa by increasing the total antioxidant capacity (T-AOC) and mitochondrial membrane potential (MMP). On the other hand, reducing free radicals induced oxidative (ROS) and per oxidative (MDA) damage to Hu ram semen.

Cryopreservation of ram semen is challenged by the high polyunsaturated fatty acid content in spermatozoa, which leads to increased oxidative stress and cellular damage. This study explored the potential of orally administered moringa oil (MO) or its microencapsulated form (MON) to protect ram spermatozoa during cryopreservation by assessing their effects on semen quality, antioxidant capacity, apoptosis, seminal metabolic enzyme activity, as well as molecular docking study. Fifteen Rahmani rams were randomly split into three groups (n = 5 per group) and fed a basal diet. The control group (CON) received 1 mL of distilled water orally, while the second and third groups received 2 mL of MO or 1 mL of MON, respectively, daily for 4 months. Semen samples were collected bi-weekly using an artificial vagina, pooled, extended and cryopreserved following standard protocols. Results demonstrated significantly higher (p < 0.05) post-thaw sperm viability, progressive motility and membrane integrity in the MO group compared to other groups after equilibration (at 5°C for 4 h), post-thawed ram semen (at 37°C for 30 s) or incubated at 37°C and 5% CO2 for 2 h. Regarding apoptotic sperm, the orally administered MO group had a significantly greater number of viable spermatozoa (p < 0.001) than the other groups. Although all treated groups had a lower percentage of early apoptosis than the control, MON administration resulted in a significant increase in the percentages of necrotic sperm compared to the MO group (p < 0.05). The TAC was highest and MDA was lowest (p < 0.05) in the MO group. Molecular docking analysis revealed the binding energies (kcal/mol) of bioactive compounds from MO including apigenin, ferulic acid and naringenin with three target proteins: ADAM17 (-4.47, -4.49 and -4.88, respectively), DNase1 (-4.42, -3.47 and -4.46, respectively) and SHBG (-6.38, -4.70 and -6.43, respectively). These findings indicate that orally administering MO has a more pronounced positive effect on Rahmani ram semen quality, apoptosis, and antioxidant status following cryopreservation compared to its microencapsulated form.

β-Carotene addition into a Tris-based diluent improves Hu ram sperm parameters after cryopreservation.

Refrigerated storage of ram sperm in presence of Trolox and GSH antioxidants: Effect of temperature, extender and storage time

Viability of bovine ivf embryos biopsied with microsection or microaspiration technique for sexing

Context The conflicting findings regarding the impact of equilibration time on post-thawed sperm quality underscore the need for further research to evaluate the impact of equilibration time and cooling rate on post-thaw sperm quality of ram semen. Aims The current study aimed to assess the combined impact of cooling rates and pre-freezing equilibration times on post-thaw sperm quality in Kail ram semen (n = 5). Methods Semen collection was performed using an artificial vagina at 42°C. The pooled semen was divided into equal aliquots and subjected to either slow cooling (SC, −0.27°C/min) or moderate cooling (MC, −0.36°C/min) rates, transitioning from 37°C to 4°C. Equilibration times of 0, 4, 8, and 12 h were employed before freezing. Key results Semen samples undergoing the SC rate and equilibrated for 4 h exhibited higher (P &amp;lt; 0.05) percentages of progressive motile (PM), rapid progressive (RP), and medium progressive (MP) sperm compared with the MC rate. However, total motility remained unaffected by the cooling rate (P &amp;lt; 0.05). Semen equilibrated for 4 h demonstrated higher (P &amp;lt; 0.05) percentages of PM and RP sperm, as well as improved kinematics (curvilinear velocity, average path velocity, and straight-line velocity) compared with other equilibration times. Nevertheless, equilibration time had no (P &amp;gt; 0.05) impact on the amplitude of the lateral head displacement for semen samples subjected to the MC rate. Notably, the cooling rate did not affect post-thaw sperm kinematics, plasma membrane integrity, or live-sperm percentage (P &amp;gt; 0.05). Semen samples equilibrated for 4, 8, and 12 h exhibited a higher (P &amp;lt; 0.05) percentage of sperm with intact plasma membrane and viability than did those equilibrated for 0 h. Conclusions In conclusion, slow cooling rate and a 4 h equilibration period were shown to be optimal for preserving post-thaw sperm quality in Kail rams. Implications The findings highlighted the combined effect of cooling rate and equilibration time on post-thaw sperm quality for optimising sperm cryopreservation protocols in the context of ram semen.

Effects of astaxanthin supplementation on the freezability, lipid peroxidation, antioxidant enzyme activities and post-thawing fertility of ram semen

Liquid nitrogen vapor is comparable to liquid nitrogen for storage of cryopreserved human sperm: evidence from the characteristics of post-thaw human sperm

This study aimed to evaluate the effect of Marjoram leaves (Oregano vulgar) as a feed supplement on sexual efficiency and semen quality in Awasian pollination rams. The study was conducted on ten rams in one of the private fields in the Babylon governorate. The rams were divided randomly into two groups. First group rams are regarded as a control group (group A) that feeds on exceptional concentric food for pollination rams. The second group consists of 5 rams (group B) provided with the same concentric food plus a supplement of 3 mg/kg of body weight of Oregano vulgar leaves (fresh) 3 times daily for 49 days before the pollination season (April and May). Semen samples were collected at the end of 49 days by artificial vagina to evaluate the volume of ejaculate, sperm number, concentration, motility, viability, and deformities, and to measure the sperm pleomorphic parameters of the head, nucleus, and acrosome. The antioxidant status of seminal plasma was evaluated by measuring superoxide dismutase (SOD), glutathione peroxidase (GPX), and total antioxidant capacity (TAC). ELISA tests were used on blood serum samples to measure the hormone levels of luteinizing hormone (LH), testosterone, and estrogen. The study demonstrated a significant increase in motility, the viability of sperm, and decreasing deformities. There was no significant variation in perimorphic parameters of the head, nucleus, and acrosome sperm after using the fresh leaves of Oregano vulgar as supplementation to Awasion sheep. However, there was an increase in the antioxidant enzymes (SOD, GPX, and TAC) in seminal plasma and an increase in LH, testosterone, and estrogen in the blood serum of experimental group B. These findings show that using fresh leaves of Oregano vulgar improved the sexual efficiency of pollinated rams and maintained the physiology and perimorphic parameters of semen and sperm.

The aim of the study was to evaluate the seasonal variation in semen quality of Dorper rams using different semen collection techniques. The study was carried out from January 2012 to January 2013. A general management programme for health control was followed, with water being provided ad libitum throughout the trial, and all rams being fed a 2.5 kg maintenance diet per day. Eleven mature Dorper rams, recording a mean body weight of 69.6 ± 9.2 kg and mean age of 18 ± 4.7 months, were used in the trial. A group of six rams were trained for semen collection with the aid of the artificial vagina (AV), while in the remaining five rams, semen was collected using the electro ejaculator (EE). Immediately after collection, ejaculates were evaluated macroscopically and microscopically for semen volume, semen colour, semen pH, semen wave motion, sperm motility, sperm cell concentration, sperm viability and morphology. The results of the trial generally showed that semen in Dorper rams may be collected using the AV or EE methods throughout the year. However, an overall significant better semen quality collected by the AV versus the EE collection method was recorded. Generally, semen of significantly higher quality was recorded in summer, autumn and spring (both collection techniques). The tendency in the current trial was that the EE technique of semen collection was the less reliable method. Consequently the AV is recommended as the more acceptable method of semen collection in the Dorper. Winter is not generally recommended for semen collection, especially when using the EE.Keywords: Artificial vagina, electro-ejaculator, season, semen quality

The protective effect of egg yolk from different avian species during the cryopreservation of Karayaka ram semen