scholarly journals The effect of L-glutamine and trehalose on dog sperm cryopreservation

2021 ◽  
Vol 90 (2) ◽  
pp. 201-206
Author(s):  
Caner Öztürk ◽  
Neşe Hayat Aksoy

This study aimed to test different doses of L-glutamine and trehalose in the canine semen diluent while determining their protective effects on spermatological and biochemical indices of the thawed samples. Semen samples were collected from three fertile dogs using the digital manipulation method. The mixed ejaculates were divided into five portions at 37 °C and diluted with additives. Five study groups were formed with L-glutamine (10 and 20 mM), trehalose (25 and 50 mM), and no additives (control). After the dilution, the semen samples were cooled for 1.5 h at 5 °C and frozen (-110 to -120 °C) in liquid nitrogen vapor. Then, they were stored at -196 °C. For spermatological evaluations, samples were thawed at 38 °C for 30 s. L-glutamine (20 mM) was found to be significantly different (P < 0.05) and led to higher percentages of motility, membrane integrity, and acrosome integrity compared to the control group. Considering the total oxidant status (TOS) assay, the lower values were determined in all the antioxidant groups compared to the control group (P < 0.05). Supplementing the semen extender with L-glutamine showed a higher total antioxidant status (TAS) concentration compared to the control group (P < 0.05). As a result of this study, a higher protective effect was found in all the spermatological evaluations after thawing the frozen semen samples, especially in the group containing L-glutamine (20 mM).

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.


2013 ◽  
Vol 2013 ◽  
pp. 1-10 ◽  
Author(s):  
P. Perumal ◽  
Kezhavituo Vupru ◽  
K. Khate

The present study was undertaken to assess the effect of melatonin (MT) on sperm motility, viability, total sperm abnormality, acrosomal and plasma membrane integrity, DNA abnormality, antioxidant profiles such as superoxide dismutase (SOD), catalase (CAT), glutathione (GSH) and total antioxidant capacity (TAC), enzymatic profiles such as aspartate amino transaminase (AST), alanine amino transaminase (ALT), and biochemical profiles such as malonaldehyde (MDA) production and cholesterol efflux. Total numbers of 30 ejaculates were collected twice a week from eight mithun bulls and semen was split into five equal aliquots, diluted with the TEYC extender. Group 1 has semen without additives (control) and group 2 to group 5 have semen that was diluted with 1 mM, 2 mM, 3 mM, and 4 mM of melatonin, respectively. These seminal parameters, antioxidant, enzymatic, and biochemical profiles were assessed at 5°C for 0, 6, 12, 24, and 30 h of incubation. Inclusion of melatonin into diluent resulted in significant (P<0.05) decrease in percentages of dead spermatozoa, abnormal spermatozoa, and acrosomal abnormalities at different hours of storage periods as compared with control group. Additionally, melatonin at 3 mM has significant improvement in quality of mithun semen than melatonin at 1 mM, 2 mM or 4 mM stored inin vitrofor up to 30 h. It was concluded that the possible protective effects of melatonin on sperm parameters are it prevents MDA production and preserve the antioxidants and intracellular enzymes during preservation.


2014 ◽  
Vol 2014 ◽  
pp. 1-9 ◽  
Author(s):  
P. Perumal

The present study was undertaken to assess the effect of superoxide dismutase (SOD) on sperm motility, and viability; total sperm abnormality; acrosomal and plasma membrane integrity; DNA abnormality; antioxidant profiles such as catalase (CAT), reduced glutathione (GSH), and total antioxidant capacity (TAC); enzymatic profiles such as aspartate amino transaminase (AST), and alanine amino transaminase (ALT); and biochemical profiles such as malondialdehyde (MDA) production and cholesterol efflux. Total numbers of 50 ejaculates were collected twice a week from eight mithun bulls and semen was split into four equal aliquots, diluted with the TEYC extender. Group 1: semen without additives (control), and group 2 to group 4: semen was diluted with 50 U/mL, 100 U/mL, and 150 U/mL of SOD, respectively. These seminal parameters, antioxidant, enzymatic, and biochemical profiles were assessed at 5°C for 1, 6, 12, 24, and 30 h of incubation. Inclusion of SOD into diluent resulted in significant (P<0.05) decrease in percentages of dead spermatozoa, abnormal spermatozoa, and acrosomal abnormalities at different hours of storage periods as compared with control group. Additionally, SOD at 100 U/mL has significant improvement in quality of mithun semen than SOD at 50 or 150 U/mL stored in in-vitro for up to 30 h. It was concluded that the possible protective effects of SOD on sperm parameters are that it prevents MDA production and preserves the antioxidants and intracellular enzymes during preservation.


2018 ◽  
Vol 19 (3) ◽  
pp. 287-291 ◽  
Author(s):  
Ashish Verma ◽  
Vivek Tripathi ◽  
Sahib T Singh ◽  
Chetan D Singh ◽  
Jaspreet S Gill

ABSTRACT Introduction Periodontitis is a common problem affecting a significant population of the world. For the assessment of oxidative stress of an individual, total oxidation status (TOS) and total antioxidant capacity (TAOC) are the significant biomarkers. Hence, we planned the present study to assess malondialdehyde (MDA), TOS, TAOC levels, and oxidative stress index (OSI) in generalized aggressive periodontitis (GP) and chronic periodontitis (CP) patients. Materials and methods The present study included assessment of 40 CP patients, 40 GP patients, and 40 healthy controls. Clinical assessment of all the subjects was done by measuring the probing depth (PD), clinical attachment (CL), gingival index (GI), gingival bleeding index (GBI), and plaque index (PI). Salivary and serum samples were taken and assessed by standard procedures as described previously in the literature. All the values were assessed and compared. Results Significant results were obtained while comparing all the periodontal parameters in between various study groups. Mean serum MDA levels in the CP, GP, and control group were found to be 0.68, 0.65, and 0.61 µM respectively. Statistically nonsignificant results were obtained while comparing the serum MDA levels in between the three study groups. Significant results were obtained while comparing the mean serum and salivary TOS values, TAOC values, and OSI in between various study groups. Conclusion In periodontitis patients, oxidative stress was significantly higher in comparison with healthy subjects. Clinical significance Oxidative parameters do play a significant role in the pathologic profile of periodontitis. How to cite this article Tripathi V, Singh ST, Sharma V, Verma A, Singh CD, Gill JS. Assessment of Lipid Peroxidation Levels and Total Antioxidant Status in Chronic and Aggressive Periodontitis Patients: An in vivo Study. J Contemp Dent Pract 2018;19(3):287-291.


2020 ◽  
Vol 32 (2) ◽  
pp. 139
Author(s):  
Z. Raphalalani ◽  
F. Ramukhithi ◽  
R. Ndhlala ◽  
K. Nephawe ◽  
T. Nedambale

The processes of semen cryopreservation and thawing affect sperm membrane integrity and motility and increases morphological defects as well as DNA damage. The most influential cause of this is oxidative stress. When endogenous antioxidant capacity of seminal plasma is reduced during the freeze-thawing process, plant extracts exhibiting strong antioxidant activity can be used as supplements for compensation. Baobab oil has gained interest because it is rich in powerful antioxidants, which could protect sperm cells from oxidative damage during cryopreservation. Our study aimed to assess the effects of baobab oil on post-thaw sperm quality parameters in an egg-yolk-based extender. Thirty semen ejaculates were collected from 15 Nguni bulls using an electro ejaculator. Semen samples were randomly allocated to control (no baobab oil), 20μL (1%), 50μL (2.5%), and 100μL (5%) baobab oil per millilitre extender. Following dilution, semen samples were loaded into 0.25-mL semen straws, equilibrated for 4h at 5°C, and transferred into a controlled rate programmable freezer. The frozen semen straws were stored in a liquid nitrogen tank (−196°C) until thawing. Semen straws were thawed (37°C/60 minutes) after 1 week of cryopreservation and analysed for (1) sperm motility using a computer-aided sperm analyser, (2) morphological defects and viability using eosin-nigrosin stain, (3) membrane integrity by hypo-osmotic swelling test, and (4) DNA integrity by terminal deoxynucleotidyl transferase dUTP nick end labelling assay. Data was analysed using analysis of variance. Treatment means were compared in relation to the control group by Dunnett's test. We found that supplementing semen extender with baobab oil at 1% significantly (P&lt;0.05) preserved sperm DNA integrity (88.3±3.7) and membrane integrity (74.0±4.2) when compared with the control group (71.7±3.7 and 55.8±4.4, respectively). Baobab oil supplementation either at 1% (5.9±0.5), 2.5% (7.2±0.5), or 5% (6.0±0.5) significantly reduced sperm morphological defects compared with control (9.5±0.5). Total motility (1% (72.7%), 2.5% (72.7%), 5% (71.9%), control (59.3%)) and viability (1% (79.1%), 2.5% (79.8%), 5% (77.8%), control (67.6%)) were also improved by supplementation; however, the difference was not significant. In conclusion, it was demonstrated that supplementing bull semen extender with 1% baobab oil protects sperm from morphological defects, maintains membrane integrity, as well as preserves sperm DNA. All the baobab oil supplementation levels preserved post-thaw bull-sperm quality parameters.


2019 ◽  
Vol 207 (3-4) ◽  
pp. 187-196
Author(s):  
Levent Tumkaya ◽  
Adnan Yilmaz ◽  
Kerimali Akyildiz ◽  
Tolga Mercantepe ◽  
Zihni Acar Yazici ◽  
...  

The use of devices, including mobile phones, generating electromagnetic fields (EMF) is widespread and is progressively increasing. It has also been shown that EMF may have detrimental effects. This is the first study to investigate the postnatal biochemical and histological effects of prenatal exposure of rat livers to 1,800-MHz EMF at different time intervals in uteroplacental life. The 3 EMF groups of rats were exposed to 1,800-MHz EMF for 6, 12, or 24 h daily for 20 days. Unexposed rats served as control group. All rats were subjected to anesthesia, and on postnatal day 60, the livers were excised, and blood was collected for histological and biochemical analyses. Malondialdehyde levels were significantly higher in the exposed groups than the unexposed controls (p < 0.05). In contrast, EMF-exposed groups had lower liver tissue glutathione levels than controls (p < 0.05). Serum Ca2+, alanine transaminase, and aspartate aminotransferase levels were higher in EMF-exposed groups than controls (p < 0.05). In addition, liver tissue total oxidant status levels were increased (p < 0.05), and liver tissue total antioxidant status levels were decreased (p < 0.05) compared to the control group. Furthermore, in the EMF groups, extensive vacuolation and degeneration of the hepatocytes in the portal area, as well as those surrounding the sinusoids, were evident. Affected hepatocytes had polygonally shaped nuclei and vacuolic cytoplasm imparting eosinophilic staining. Loss of cellular membrane integrity and invaginations, as well as picnotic nuclei, was prominent. This study has shown that intrauterine liver damage caused by 1,800-MHz EMF exposure persists into puberty in rats.


Author(s):  
N. N. Malyutina ◽  
A. F. Bolotova ◽  
R. B. Eremeev ◽  
A. Zh. Gilmanov ◽  
D. Yu. Sosnin

Introduction. The overwhelming number of publications contains only data on the content of individual antioxidants, but not on the overall antioxidant activity of the blood in patients with vibration disease.The aim of the study was to determine the total antioxidant activity of blood serum in patients with vibration disease.Materials and methods. Th e main group consisted of 30 people diagnosed with “Vibration disease” of 1 degree (n=21) and 2 degrees (n=9). Th e control group consisted of 30 clinically healthy men, comparable in age with the main group (p=0.66). Th e total activity of antioxidant systems of blood plasma was evaluated photometrically using the test system “Total antioxidant status-Novo” (“Vector-best”, Russia).Results. The indicator of the total antioxidant status (TAS) was 1,038±0.232 mmol/l in the examined main group, against 1,456±0.225 mmol/l in the examined control group (p<0.000001). Th e coefficient of variation (CV) in patients with vibration disease was 22.35%, 1.45 times higher than in the control group (15.45%). In the main group there was a positive correlation between age and TAS (R=0.525), in the control group there was no such relationship (R=0.095). Th e degree of decrease depended on the severity of vibration disease.Conclusions. 1. The development of vibration disease is accompanied by a decrease in the antioxidant status of blood serum. 2. Th e degree of decrease in the antioxidant status of blood serum correlates with the severity of vibration disease. 3. Reduction of TAS can serve as a pathogenetic justification of the need to include drugs and/or biologically active additives with antioxidant activity in therapy


2017 ◽  
Vol 28 (01) ◽  
pp. 096-100
Author(s):  
Leyla Tekin ◽  
Mehmet Erdemli ◽  
Nazile Erturk ◽  
Zeynep Aksungur ◽  
Serdar Aktas ◽  
...  

Purpose This study aimed to compare the protective effects of Hypericum perforatum (Hp) and quercetin, a flavonoid, against ischemia/reperfusion (I/R) injury in rat testes. Materials and Methods This study included 28 male Wistar albino rats that were divided into four groups. Except for the sham group, torsion was created by rotating both testes at an angle of 720 degrees clockwise for 2 hours. The Hp and quercetin groups received 25 mg/kg Hp and quercetin intraperitoneally 30 minutes before detorsion, respectively. Orchiectomy was performed for the measurement of markers of oxidative stress and histopathological examination. Results In the Hp and quercetin groups, malondialdehyde (MDA) and nitric oxide (NO) levels and total oxidant capacity were significantly lower, the glutathione level and total antioxidant status were significantly higher, and Johnsen's testis biopsy scores were significantly higher than in the torsion/detorsion group (p ˂ 0.001). The markers of oxidative injury were significantly lower (p ˂ 0.001) and total antioxidant status was significantly higher (p ˂ 0.001), except for glutathione (p = 0.62) in the Hp group than in the quercetin group. Johnsen's score between Hp and quercetin groups was not significantly different (p = 0.80). Conclusion Both Hp and quercetin have protective effects against I/R injury of the testes, but the protective effect of Hp was found to be stronger than that of quercetin.


2007 ◽  
Vol 61 (1) ◽  
Author(s):  
K. Pawłowska-Góral ◽  
M. Wardas ◽  
P. Wardas ◽  
A. Rzepecka-Stojko

AbstractThe influence of CCl4 on the activity of superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione reductase (GR), the value of the total antioxidant status (TAS), and the concentration of malonic dialdehyde (MDA) and glutathione (GSH) was monitored in plasma or whole blood of rabbits. The administration of CCl4 caused the increase of the SOD activity to approximately 150 % and the decrease in the activity of GPx and GR by about 50 %. These changes were accompanied with the increase in TAS value and MDA concentration and the decrease of GSH concentration. The effect of CCl4 was suppressed by the previous 7 days lasting or simultaneous administration of vitamin E. Oxidative stress caused by CCl4 was accompanied by the development of reactive oxygen forms, especially superoxide radical anion.


2011 ◽  
Vol 23 (1) ◽  
pp. 110
Author(s):  
M. A. Coutinho da Silva ◽  
C. R. F. Pinto ◽  
J. M. Young ◽  
K. Cole

Magnetic-activated cell sorting (MACS) has been used successfully in humans to remove apoptotic sperm from the ejaculate. Annexin V-conjugated microbeads recognise sperm with externalized phosphatidylserine, which is considered one of the features of apoptosis, and the labelled sperm is separated by MACS. The goals of the study were to determine if MACS can be used to separate apoptotic sperm from the ejaculate of stallions; and to determine if removal of apoptotic sperm improves the quality of stallion sperm. Our hypothesis was that MACS would improve semen quality by removing apoptotic sperm, resulting in samples with higher motility and viability. Two ejaculates from three different stallions of good fertility were used. Sperm were diluted with Tyrode’s albumin lactate pyruvate (TALP) and incubated with annexin V-conjugated microbeads for 15 min at 37°C. Control samples were incubated in the absence of annexin V microbeads. The suspension was then loaded into the separation column containing iron globes, which were fitted in a magnet (MiniMACS; Miltenyi Biotec Inc., Auburn, CA, USA). The effluent sample containing annexin-negative sperm was collected and then, the column was removed from the magnetic field and rinsed with TALP to collect the annexin-positive cells. Sperm viability, motility, morphology and caspase activation were determined in all three samples: control, annexin-negative, and annexin-positive. Data were evaluated by ANOVA and individual comparisons were performed by Tukey’s hsd test. Significance was set at P < 0.05 and data is presented as means ± SEM (Table 1). The main effect of stallion was significant only for sperm motility parameters. Sperm recovery rate following MACS was 46 ± 3%. In conclusion, the use of MACS was effective in removing apoptotic sperm from the ejaculate. The annexin-positive population displayed a higher proportion of sperm with activated caspases and lower membrane integrity and motility. However, removal of apoptotic sperm from the ejaculate did not improve sperm parameters in the annexin-negative group compared to control group. In addition, sperm morphology was not affected by MACS. Further studies are necessary to determine if MACS could be used successfully to improve sperm quality from subfertile stallions and frozen semen. Table 1.Sperm parameters following annexin V MACS (mean ± SEM) The authors are thankful to Mark Williams at Miltenyi Biotec Inc. for providing supplies; and Dr Ashok Agarwal at The Center for Reproductive Medicine, Cleveland Clinic, for scientific input.


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