scholarly journals Chlorogenic Acid Improves Quality of Chilled Ram Sperm by Mitigating Oxidative Stress

Animals ◽  
2022 ◽  
Vol 12 (2) ◽  
pp. 163
Author(s):  
Yanhu Wang ◽  
Liuming Zhang ◽  
Tariq Sohail ◽  
Yan Kang ◽  
Xiaomei Sun ◽  
...  

The purpose of this study was to investigate whether the addition of chlorogenic acid (CGA) to a sheep semen extender could improve the quality of chilled sheep sperm. Ejaculates (n = 80) were collected from five Hu rams with an artificial vagina. The ejaculates were mixed and divided into five equal parts, diluted with a CGA-free Tris–egg yolk extender (control), or supplemented with 0.2, 0.4, 0.8, and 1.2 mg/mL. The sperm kinematic parameters (viability, progressive motility), functional integrity of plasma membrane and acrosome, adenosine triphosphate (ATP) concentration and antioxidant parameters (Catalase (CAT), Superoxide dismutase (SOD) activity, total antioxidant capacity (T-AOC), ROS level and Malondialdehyde (MDA) content) were evaluated during storage of the semen. The results indicated that: PM, plasmatic membrane integrity and acrosomal integrity in 0.8 mg/mL CGA were higher (p < 0.05) from day 1 to 5. The ROS level in CGA groups was lower than the control (p < 0.05). CAT, SOD, ATP, and T-AOC were highest at 0.8 mg/mL concentration within 1 to 5 days. The above results indicated that the right concentration of CGA improved the quality of Hu ram sperm during chilling storage.

Animals ◽  
2020 ◽  
Vol 10 (9) ◽  
pp. 1508 ◽  
Author(s):  
Sameh Abdelnour ◽  
Mahmoud Hassan ◽  
Amer Mohammed ◽  
Ahmad Alhimaidi ◽  
Naif Al-Gabri ◽  
...  

The cryopreservation process adversely affects sperm function and quality traits, causing some changes at biochemical and structural levels, due to mechanical, thermal, osmotic, and oxidative damage. Supplementation with curcumin nanoparticles could prevent and even revert this effect and could enhance the post/thawed sperm quality in the rabbit. The study amid to explore the effect of curcumin (CU) and curcumin nanoparticles (CUNPs) supplementation in semen extender on post/thawed rabbit sperm quality. Twelve fertile, healthy rabbit bucks were included, and the ejaculates were collected using artificial vaginas. Rabbit pooled semen was cryopreserved in tris-yolk fructose (TYF) extender without any supplement (control group) or extender supplemented with CU at levels of 0.5, 1 or 1.5 µg/mL (CU0.5, CU1.0, and CU1.5, respectively) or CUNPs at levels of 0.5, 1, 1.5 (CUNPs0.5, CUNPs1.0, and CUNPs1.5, respectively) and was packed in straws (0.25 mL) and stored in liquid nitrogen (−196 °C). Results revealed that CUNPs1.5 had a positive influence (p < 0.05) on post-thawing sperm progressive motility, viability, and membrane integrity as compared with the other groups. Percentages of dead sperm, abnormalities, early apoptotic, apoptotic, and necrotic sperm cells reduced (p < 0.05) in CUNPs1.5 as compared to other treatments. Using 1.5 µg/mL of CUNPs significantly improved total antioxidant capacity (TAC), GPx, while MDA and POC reduced (p < 0.05) in CU1.5 in comparison with other groups. SOD values were enhanced (p < 0.05) in CUNPs1.0 and CUNPs1.5 in relation with other treatments. Conclusively, the addition of curcumin and its nanoparticles to the extender can improve the post-thawed quality of rabbit sperm via redox signaling and reduce the apoptosis process.


Author(s):  
Jiří Šichtař ◽  
Ondřej Šimoník ◽  
Petra Folková ◽  
Adéla Dokoupilová ◽  
Radko Rajmon ◽  
...  

The aim of this study was to evaluate the effect of clarified egg yolk addition to semen extender, and the semen collection sequence on the quality of frozen-thawed semen in dogs. Semen was collected from 6 dogs in a time interval of 24 hours. As parameter of the quality of frozen-thawed (F-T) semen, the motility by computer assisted sperm analysis (CASA) and plasma membrane integrity by hypo-osmotic swelling test (HOS) were evaluated. All kinematic parameters of sperm motility were higher in F-T samples containing the whole in comparison to the clarified egg yolk. The sequence of semen collection affected sperm movement characteristics of native as well as F-T semen, but it was not possible to determine whether the fresh semen from the 1st or 2nd collection is of higher quality. All motility parameters of sperms frozen with extender containing the whole egg yolk were significantly higher in the case of the 2nd collection. The situation was not so clear in the case of clarified egg yolk addition, but the velocity values were higher in F-T samples from the 2nd collection. In contrast to proven differences in motility, the effect of the addition of clarified egg yolk and the sequence of semen collection were not projected at all on the quality of plasma membrane of canine sperms evaluated by HOS test.


2022 ◽  
Vol 43 (2) ◽  
pp. 841-854
Author(s):  
Lucas Emanuel Ferreira Canuto ◽  
◽  
Lorenzo Garrido Teixeira Martini Segabinazzi ◽  
Endrigo Adonis Braga de Araújo ◽  
Luis Fernando Mercês Chaves Silva ◽  
...  

Cooling and freezing processes cause physical and chemical damage to sperm by cold shock and oxidative stress. This study aimed to evaluate the effect of two antioxidants on sperm parameters of cooled and frozen-thawed ram semen diluted in an egg yolk-based extender. Semen was collected from 30 rams and processed in two consecutive experiments to test the inclusion of different concentrations of quercetin and butylated hydroxytoluene (BHT) in an egg yolk-based semen extender. Dimethyl sulfoxide (DMSO) was added as a solvent to the semen extender in a ratio of 1 mL DMSO for 90 mg of quercetin and 1 mL DMSO for 880 mg of BHT. After collection, semen was diluted at 200 × 106 motile sperm/mL (control) and split into different groups in each experiment. In experiment 1, semen was diluted with the extender containing quercetin (Q5, 5 μg/mL; Q10, 10 μg/mL; Q15, 15 μg/mL) or DMSO alone (DMSO1, 0.055 μL DMSO per mL; DMSO2, 0.165 μL DMSO per mL). In experiment 2, semen was diluted with the extender with BHT (BHT1, 0.5 μg/mL; BHT2, 1 μg/mL; BHT3, 1.5 μg/mL) or DMSO alone (DMSO3, 0.375 μL DMSO per mL; DMSO4, 1.125 μL DMSO per mL). After dilution, the semen was divided into two aliquots. Treated ram sperm samples were also subjected to different storage methods. The first set of samples was cooled at 5 °C for 24 h, whereas the second set of samples was frozen-thawed. Sperm motility parameters and plasma membrane integrity (PMI) were evaluated immediately after dilution (0h) and 24 h after cooling and in the frozen-thawed samples via computer-assisted sperm analysis and epifluorescence microscopy, respectively. The inclusion of quercetin or BHT did not affect sperm motility parameters or PMI of fresh, cooled, or frozen-thawed sperm in this study (P < 0.05). However, further studies are needed to test the effects of these antioxidants on the fertility of cryopreserved ram semen.


2020 ◽  
Vol 7 (2) ◽  
pp. 235-241
Author(s):  
Pankaj Kumar Jha ◽  
M Golam Shahi Alam ◽  
Farida Yeasmin Bari

The effect of freezing methods and diluents types on post-thaw sperm quality of Bangladeshi ram semen was studied. Two freezing methods and three diluents was tested as pooling effects (freezing methods or diluents) on post-thaw sperm parameters; sperm motility (SM), viability (SV), plasma membrane integrity (SPMI) and acrosome integrity (SAI), respectively. From selected ten rams, eight ejaculates were used for each freezing group (freezing methods × diluents). Semen samples were diluted by using two-steps for hand-made tris-based diluents (20% egg yolk): D1 (7% glycerol) and D2 (5% glycerol), and one-step dilution for commercial diluents: D3 (Triladyl®) at 35°C. After 4h of equilibration of temperature at 5°C, diluted semen samples was aspirated into 0.25 mL straws, and sealed. Straws were frozen in liquid nitrogen (LN2) vapour using two methods: F1 (manually in Styrofoam box, using three-steps method; +5°C to -80°C at -11.33°C/min, -80°C to -120°C at -26.66°C/min, and -120°C to -140°C at - 13.33°C/min) and F2 (programmable bio-freezer, using two-steps method; +5°C to -100°C at - 20°C/min and -100°C to -140°C at -10°C/min). Two semen straws from each batch were evaluated (37°C for 20 sec) for sperm parameters. In pool effects between freezing methods; SAI differed significantly (P < 0.001). The SM (56%) and SV (72%) were observed competitive. However, SPMI (67.58 ± 2.02%) and SAI (76.13 ± 1.42%) were higher in F1. Among diluents, SM (P < 0.006), SV (P < 0.008), SPMI (P < 0.012) and SAI (P < 0.019) differed significantly. The SM (61.25 ± 1.80%), SV (77.13 ± 1.47%), SPMI (68.31 ± 1.91%) and SAI (74.75 ± 1.64%) were highest in D3. In conclusion, the combination of manual freezing (three-steps) and handmade tris-based diluents (20% egg yolk, 5% glycerol) is suitable and sustainable method for cryopreservation of ram semen. Res. Agric., Livest. Fish.7(2): 235-241,  August 2020


2020 ◽  
Vol 32 (2) ◽  
pp. 145
Author(s):  
C. Souza ◽  
F. Brandão ◽  
J. Santos ◽  
V. Alfradique ◽  
V. Brair ◽  
...  

The cryopreservation process causes oxidative stress to the sperm cell, and the addition of antioxidants to the extender for semen freezing helps sperm protection. This study assessed the effect of L-carnitine (LC) concentrations (0, 5, or 10mM LC) on two ram semen extenders (Tris-egg yolk or the commercial optiXcell IMV medium (IMV Technologies)) for semen cryopreservation. Four Santa Inês rams were used during the breeding season. After semen collection, macroscopic and microscopic evaluations were performed, and a pool of semen was formed. The semen was diluted, and the final concentration was 100×106 per 0.25-mL straw. Cryopreservation was performed with a cooling rate of 0.25°C min−1 until 5°C, and the freezing rate used was 20°C min−1 from 5 to −120°C. After the freezing-thawing process and throughout incubation (38°C in 5% CO2) in Fert-Tyrode's albumin lactate pyruvate medium, every 1h for up to 3h, several parameters were evaluated: sperm kinetics, hypo-osmotic test, plasma membrane integrity, capacitation status, and lipid peroxidation level. We did not find any protective effect of LC on plasma membrane integrity, hypo-osmotic test, and capacitation status. The sperm kinetics values throughout incubation showed that Tris extender promoted better indices of staight-line velocity, linearity, wobble, and straightness than IMV extender along incubation, regardless of the presence of LC. There were no benefits of the LC addition throughout the incubation, and 10mM was deleterious to few parameters (amplitude of lateral head displacement, linearity, and wobble) compared with the control (0mM) in the Tris extender group. The plasma membrane integrity analysis revealed no differences (P&gt;0.05) among the groups. The average number of intact cells (hypo-osmotic) was higher in Tris extender groups supplemented with 10mM LC at 1h and 5mM LC at 2h compared to the respective extender IMV groups. Regarding capacitation status, the Tris 5mM LC group had more acrosome-reacted cells when compared with the IMV 5mM LC group at 2h. At 3h, the percentage of acrosome-reacted cells was higher in the Tris 0-mM group when compared with the IMV 0-mM group. Regardless the presence of LC, IMV had higher (P&lt;0.05) lipoperoxidation than the Tris treatments. In conclusion, LC supplementation in semen extender had no beneficial effect on freezing-thawing ram sperm and throughout incubation for up to 3h, with no difference in each time point evaluated. Under the conditions of this study, the use of Tris extender was superior to IMV extender for ram sperm. Financial support for this work came from the Fundação de Amparo à Pesquisa do Estado do Rio de Janeiro (Young Scientist Program of Our State; E-26/203.168/2017).


2012 ◽  
Vol 57 (No. 8) ◽  
pp. 377-381 ◽  
Author(s):  
M. Ziaullah ◽  
A. Ijaz ◽  
M. Aleem ◽  
A.K. Mahmood ◽  
H. Rahman ◽  
...  

The study was conducted to evaluate the potential cryoprotective effect of butylated hydroxytoluene (BHT) through post-thaw evaluation of canine semen and its optimal inclusion level. Ejaculated canine semen was extended in TRIS-glucose egg yolk extender containing various concentrations of BHT (0.5, 1.0, 1.5, 2.0, and 2.5mM). Semen was frozen at &minus;196&deg;C using 200 &times; 10<sup>6 </sup>spermatozoa per 0.5 ml straws and post-thaw evaluation was carried out in terms of sperm motility, viability, plasma membrane integrity, and acrosomal integrity through phase-contrast microscope, supravital staining, hypo-osmotic swelling test, and normal acrosomal ridge, respectively. BHT was found to improve (P &gt; 0.005) all post-thawed semen quality parameters at an inclusion level of 1.0mM in the extended semen. However, higher concentrations than this were found to have detrimental effects. &nbsp;


2016 ◽  
Vol 47 (2) ◽  
pp. 60-67
Author(s):  
P. Folková ◽  
J. Šichtař ◽  
O. Šimoník ◽  
A. Dokoupilová ◽  
R. Rajmon

Abstract The aim of the study was to evaluate the effect of repeated semen collection and the substitution of normal egg yolk with clarified egg yolk to commercially produced semen extender on qualitative parameters of frozen-thawed canine semen. Two semen collections were scheduled in a 24-hour interval and in each of six dogs, three 1st and three 2nd collections were performed. The frozen-thawed sperm samples were prepared either with clarified or normal egg yolk and motility and viability were evaluated. The effect of the sequence of semen collection was demonstrated by significant differences in motility and also in viability of sperms both in native and frozen-thawed ejaculate. The percentage of viable sperms was significantly higher in samples from the 2nd compared to the 1st collection. This trend was the same also in motility except in native ejaculate. The addition of clarified egg yolk was beneficial for higher survival of sperms immediately after thawing and also after 30 min of incubation, compared to samples with normal egg yolk. Sperm motility evaluated after thawing was higher in samples with clarified egg yolk, without an apparent connection with semen collection sequence. The decrease of values of the qualitative parameters of sperms observed in the period of 30 min of incubation was significantly slowed down when clarified egg yolk was used. This was especially obvious in samples from the 2nd collection.


Author(s):  
M.M. AYBAZOV ◽  
◽  
A.N. SHEVCHENKO ◽  
M.I. SELIONOVA ◽  
T.V. MAMONTOVA

Numerous studies have proved the necessity of egg yolk in synthetic media to dilute semen before its cryopreservation. However, at the same time, it has been demonstrated that its use can adversely affect the quality of frozen-thawed sperm. The present study compares the main quality parameters of ram sperm frozen using TRIS-based diluent with egg yolk and two egg yolk-free diluents (OvixCell® and AndroMed®). A slower deterioration in the kinematic performance of sperm cryopreserved in TRIS diluent with native egg yolk confirmed higher cryoprotective performance compared to commercial extenders containing no egg yolk. Significantly higher total and progressive motility was observed in TRIS-based medium with egg yolk (P< 0.05). This advantage was maintained after four hours of incubation and became more significant at the end of cultivation (after six hours) (P< 0.01). Thus, ram sperm frozen in egg yolk medium retained better motility than in egg yolk-free extenders, which allows predicting its higher bioavailability. Assessment of some semen parameters using CASA showed that there were no significant differences in motility between the three extenders immediately after thawing the straws. When assessed two hours after thawing, a diluent containing egg yolk (TRIS-based) was found to have higher results for some of the examined traits than phospholipid diluents.


2018 ◽  
Vol 51 (1) ◽  
pp. 21-28 ◽  
Author(s):  
Sakirat Opeyemi Adeyanju ◽  
James Olatinbo Daramola ◽  
Jimoh Alao Olanite ◽  
Olufiropo Samson Awokola

Abstract Soybean lecithin had been used as an alternative to egg yolk in domestic animal semen extender during cryopreservation due to its characteristic phospholipid content which played a major cryoprotective role. This composition of soybean lecithin informed the replacement of soybean with sunflower lecithin (SL) in the extender for the Kalahari Red (KR) buck semen cryopreservation in this study. Effect of different levels of SL on the quality of the KR buck semen during cryopreservation using slow freezing method was evaluated. Semen samples were collected from four KR bucks of between two and two and half of age using artificial vagina, evaluated for motility and then diluted in extenders containing different levels of SL (1.5%, 3.0% and 4.5%) as experimental group and 0% SL or 20% egg yolk as control. Semen parameters including motility, acrosome integrity (AcI), membrane integrity (MI), malondialdehyde (MDA) concentration, cholesterol level and seminal arginase activity were evaluated for. The results showed that motility, acrosome integrity (AI) and membrane integrity were comparable at 0%, (22.00 ± 4.58, 82.00 ± 3.51 and 96.00 ± 2.03); 1.5%, (23.00 ± 2.08, 87.00 ± 3.79 and 89.00 ± 2.08); 3.0%, (13.00 ± 2.52, 81.33 ± 0.41 and 76.67 ± 1.20) and 4.5% (11.00 ± 4.51, 85.33 ± 9.88 and 84.00 ± 8.50), respectively, after thawing. SL at 0% had the highest (P < 0.05) values for MDA, cholesterol and seminal arginase activity (1.10 ± 0.008 nmol/ml, 236.35 ± 4.08 mg/dl and 0.54 ± 3.3 E-3 units/mg protein, respectively). Our data suggest that 1.5% sunflower lecithin can be used in place of soy lecithin as a substitute for egg yolk during the cryopreservation of caprine semen.


2014 ◽  
Vol 26 (1) ◽  
pp. 145
Author(s):  
K. Ogata ◽  
B. Sarentonglaga ◽  
M. Yamaguchi ◽  
A. Sasaki ◽  
Y. Kato ◽  
...  

Trans-cervical insemination (TCI) with cryopreserved semen offers a potentially effective approach for breeding canids with specific genetic traits, such as guide dogs for the blind. However, there are technical difficulties in canine sperm cryopreservation, such as the composition of semen extender. The aim of this study was to evaluate the effects of glutathione (GSH) as an antioxidant in the semen extender to improve the quality of frozen-thawed dog sperm. A Tris-egg yolk-citrate extender containing 15.7 mg mL–1 of TRIS, 8.8 mg mL–1 of citric acid, 14.1 mg mL–1 of lactose, 25.4 mg mL–1 of raffinose, 1% (vol/vol) antibiotics, and 20% (vol/vol) egg yolk in ultra-pure water was used as the base medium. Twelve ejaculates were collected from 7 dogs. Each ejaculate was divided into 2 to 5 aliquots and extended with base extender supplemented with 0, 2.5, 5, 7.5, and 10 mM GSH as first dilution. The extended semen was equilibrated for 3 h at 4°C. An equal volume of second extender was added to obtain a final concentration of 6.5% glycerol and sperm per milliliter. The sperm samples were loaded in straws and frozen at 6 cm above the surface of LN2 for 15 min in a styrene foam box and plunged into the LN2. The frozen semen was thawed for evaluation. The motility of sperm was estimated with a phase-contrast microscope and the motile patterns were classified into the following grades: progressively motile at a high speed (+++), progressively motile at a moderate and low speed (++), motile without progression (+), and immotile (–). Then, the sperm motility index (SMI) was determined from the following formula as described previously (Iritani et al., 1975), with some modifications: the percentage of (+++) sperm + the percentage of (++) sperm × 0.75 + the percentage of (+) sperm × 0.5. Sperm motility and the SMI were determined at 0, 1, 2, 3, 4, 12, and 24 h after thawing. Acrosome status was evaluated at 4 h after thawing. Lipid peroxidation (LP) levels at 0 and 12 h after thawing were used to examine the antioxidant ability of GSH. Trans-cervical insemination was carried out on 5 bitches to evaluate the fertility of GSH-treated sperm. The TCI were performed nonsurgically with a laparoscope and deposited 2 mL of semen through a catheter. Each bitch was inseminated 1 to 2 times during oestrus. Data were analysed using ANOVA with the Tukey-Kramer method. We found that the rate of (+++) sperm in the 5 mM GSH group was higher than that in the 0 mM group from 1 to 24 h after thawing (P < 0.05). The SMI was higher in the 5 and 7.5 mM GSH groups than in the 0 mM group (P < 0.05). There were no significant differences in the control and 2.5 and 10 mM GSH groups. Long-term survival was increased in the 5 mM GSH group. Acrosome integrity was higher in the GSH-treated group. The level of LP was lower in the GSH-treated groups at 0 h after thawing (P < 0.05). Trans-cervical insemination with the 5 mM GSH-treated semen resulted in the delivery of 5 pups from 2 bitches. These results indicate that the cryopreservation with 5 mM GSH can improve the motility, viability, and fertility of frozen-thawed canine sperm by its antioxidant effects on the sperm membrane.


Sign in / Sign up

Export Citation Format

Share Document