Partial Deoxygenation of Semen Extender Minimizes Post-thaw Damages and Improves Freezability of Crossbred Bull Spermatozoa

2021 ◽  
Author(s):  
Lhendup Bhutia ◽  
Abhishek Kumar ◽  
Rahul Katiyar ◽  
Vinod Gupta ◽  
M. Ramamoorthy ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Semen Quality ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Control Group ◽  
Progressive Motility ◽  
Group I ◽  
Group Ii ◽  
Semen Extender ◽  
Acrosomal Integrity

Background: Oxidative stress occurs when oxygen or oxygen derived oxidants exceed antioxidants and become responsible for poor post-thaw semen quality during the cryopreservation process. Therefore, the present study was designed to investigate the effects of dissolved oxygen (DO) levels (4, 6 and 8 ppm) in semen extender on freezability of crossbred bull spermatozoa. As the gap between 4 and 8 ppm existed in earlier studies, DO of 6 ppm in semen extender was further standardized and the effects were compared with other respective groups to contemplate any improvement in post-thaw semen quality.Methods: For the experiment, Tris-egg Yolk-Glycerol (TYG) extender was partially deoxygenated by nitrogen gassing @ 2-3 bubbles/sec at 34°C for 0, 16, 12 and 9 minutes to obtain DO levels of 11.7 ppm (Group-I/control), 4 ppm (Group-II), 6 ppm (Group-III) and 8 ppm (Group IV), respectively and collected semen samples were diluted with these extender groups to have 80×106 spermatozoa/ ml of the extender. Semen samples were evaluated for individual progressive motility (IPM), lipid peroxidation (LPO), total antioxidant capacity (TAC), plasma membrane integrity, acrosomal integrity and apoptotic changes at different stages of cryopreservation.Result: At the post-thaw stage, progressive motility was greater (p less than 0.05) in Group II compared to Group I and the least reduction from the post-dilution to the post-thaw stage was observed in Group II. In comparison to Group I, Groups II, III and IV showed lesser (p less than 0.05) MDA production with Group II having greater (p less than 0.05) TAC concentration than other groups at the post-thaw stage. A declining trend was observed in membrane integrity as DO levels increased from 4 ppm to 11.7 ppm. Acrosomal integrity did not differ among treatment groups, but, found to be higher (p less than 0.05) than the control group. Per cent viable spermatozoa was greater (p less than 0.05) in Group II than Group I and vice versa for necrotic spermatozoa as assessed by Annexin VFITC/PI staining. In conclusion, reducing the DO level to 4 ppm before cryopreservation improved the freezability by reducing oxidative stress and apoptotic changes while, above 4 ppm tended to lower it. An appreciable improvement in freezability can be seen at 6 ppm of DO, but, not up to that extent as observed at 4 ppm.

Effect of optimization of the levels of dissolved oxygen in semen extender on physico-morphological attributes and functional membrane integrity of crossbred bull spermatozoa

2018 ◽  
Author(s):  
Abhishek Kumar ◽  
J. K. Prasad ◽  
A. R. Mustapha ◽  
R. Mustapha ◽  
G. K. Das ◽  
...  
Keyword(s):  
Dissolved Oxygen ◽  
Semen Quality ◽  
Membrane Integrity ◽  
Control Group ◽  
Morphological Attributes ◽  
Progressive Motility ◽  
Group I ◽  
Functional Membrane ◽  
Group Ii ◽  
Semen Extender

The objective of present study was to investigate the effects of two different levels of dissolved oxygen (DO) i.e. 4 and 8 ppm in the extenders on physico-morphological attributes and functional membrane integrity of crossbred bull spermatozoa following cryopreservation. Only those ejaculates that had a mass activity of ³ 3+ and individual progressive motility of ³ 70% were utilized. Each of the ejaculates were divided and extended to have 80×106 spermatozoa/ mL of extender in three groups i.e. Group I (control), Group II (4 ppm) and Group III (8 ppm). Progressive motility (%) was higher (p Lass Than 0.05) in Group II and III as compared to Group I at both pre-freeze and post-thaw stage. Per cent live sperm and HOST reactive sperm were higher (p Lass Than 0.05) in Group II and in Group II and III as compared to Group I at pre-freeze and post-thaw stage, respectively. Sperm abnormality (%) was lower (p Lass Than 0.05) in Group II at post-thaw stage. Per cent acrosome intact sperm did not differ (p>0.05) among the groups at pre-freeze stage, but, were higher (p Lass Than 0.05) in Group II and III as compared to Group I at post-thaw stage. In conclusion, cryopreservation of semen having DO level of 4 ppm in semen extender yielded better results followed by 8 ppm in terms of semen quality parameters and functional membrane integrity of crossbred bull spermatozoa.

The influence of injectable sodium selenite on semen characteristics and testosterone concentration in Aardi goats

2015 ◽  
Vol 49 (6) ◽  
Author(s):  
A. Hajalshaikh ◽  
M. J. Al-Hassan ◽  
H. E. Mohamed
Keyword(s):  
Body Weight ◽  
Sodium Selenite ◽  
Semen Quality ◽  
Sperm Count ◽  
Plasma Concentrations ◽  
Control Group ◽  
Progressive Motility ◽  
Group I ◽  
Semen Characteristics ◽  
And Control

The objective of this study was to test the effects of sodium selenite (SS) injection on semen characteristics and testosterone plasma concentrations in male Aardi goats. Fifteen animals were assigned into three groups; control (group I, with no supplemental SS); group II; injected intramuscular with 0.1 mg/kg SS body weight; group III injected 0.05 mg/kg SS body weight. Blood samples were collected once weekly (week 0 and four weeks after treatment) in the morning (at 8 am). Semen was collected by electro-ejaculator on a weekly basis post SS supplementation. Sperm count, motility %, progressive motility characteristic; average path velocity (VAP), straight line velocity (VSL), curvilinear velocity (VCL), straightness (STR) and linearity (LIN) were analyzed. Significant differences were found between the two treated-groups and control after week four for motility and progressive motility. After five weeks, significant differences were found between groups, for motility and progressive motility. The live % showed significant differences after five weeks between treated and control groups. After five weeks, significant differences were found between the three groups, for motility and progressive motility. The live % showed significant differences after 5 weeks between different treatments. However, VAP, VSL, VCL, STR, and LIN showed non-significant differences, suggesting that bucks might already have a good motility characteristic. No significant effects of SS supplementation on testosterone plasma levels, and the reported values were 1.28; 1.35, and 1.36 ng/ml for control; group 1, and group 2; respectively. In conclusion, SS improved reproduction in goats <italic>via</italic> the enhancement of semen quality in Aardi goats.

scholarly journals Methyl Gallate Attenuates Doxorubicin-Induced Cardiotoxicity in Rats by Suppressing Oxidative Stress

Scientifica ◽  
2021 ◽  
Vol 2021 ◽  
pp. 1-12
Author(s):  
Akheruz Zaman Ahmed ◽  
Shakta Mani Satyam ◽  
Prakashchandra Shetty ◽  
Melanie Rose D’Souza
Keyword(s):  
Oxidative Stress ◽  
Normal Control ◽  
Wistar Rats ◽  
Group Iv ◽  
Control Group ◽  
Methyl Gallate ◽  
Group Iii ◽  
Group I ◽  
Female Wistar Rats ◽  
Group Ii

Doxorubicin-induced cardiotoxicity is the leading cause of morbidity and mortality among cancer survivors. The present study was aimed to investigate the cardioprotective potential of methyl gallate; an active polyphenolic nutraceutical, against doxorubicin-induced cardiotoxicity in Wistar rats. Twenty-four female Wistar rats (150–200 g) were divided into four groups (n = 6) which consist of normal control (group I), doxorubicin control (group II), test-A (group III), and test-B (group IV). Group III and group IV animals were prophylactically treated with methyl gallate 150 mg/kg/day and 300 mg/kg/day orally, respectively, for seven days. Doxorubicin (25 mg/kg; single dose) was administered through an intraperitoneal route to group II, III, and IV animals on the seventh day to induce acute cardiotoxicity. On the 8th day, besides ECG analysis, serum CK, CK-MB, LDH, AST, MDA, and GSH were assayed. Following gross examination of isolated hearts, histopathological evaluation was performed by light microscopy. A significant ( p  < 0.05) cardiac injury, as well as oxidative stress, was observed in doxorubicin control rats in comparison to normal control rats. Methyl gallate at both the doses significantly ( p  < 0.05) reduced doxorubicin-induced ECG changes, dyslipidaemia, and elevation of CK, CK-MB, LDH, AST, MDA and increased GSH level. Methyl gallate reversed the doxorubicin-induced histopathological changes in the heart. The present study revealed that methyl gallate exerts cardioprotection against doxorubicin-induced cardiotoxicity in female Wistar rats by suppressing oxidative stress. Our study opens the perspective to clinical studies for consideration of methyl gallate as a potential chemoprotectant nutraceutical in the combination chemotherapy with doxorubicin to limit its cardiotoxicity.

scholarly journals Effect of Curcumin on Oxidative Stress in a Model of Turpentine Induced Acute Experimental Inflammation

2017 ◽  
Vol 45 (1) ◽  
pp. 65-74 ◽  
Author(s):  
Andrei CONEAC ◽  
Meda Sandra ORASAN ◽  
Daniel Corneliu LEUCUTA ◽  
Nicoleta DECEA ◽  
Miuta FILIP ◽  
...  
Keyword(s):  
Oxidative Stress ◽  
Antioxidant Capacity ◽  
Total Antioxidant Capacity ◽  
High Performance ◽  
Control Group ◽  
Group Iii ◽  
Group I ◽  
Total Antioxidant ◽  
Experimental Inflammation ◽  
Group Ii

Curcumin, a natural phenolic compound is an anti-tumor agent with anti-inflammatory and anti-oxidant properties. The aim of this research was to evaluate oxidative stress levels, the antioxidant activity and Curcumin concentrations by high performance liquid chromatography (HPLC) in an acute experimental inflammation induced by Turpentine oil (intramuscular 0.6 mg kg-1 body weight) and to compare a prophylactic versus a therapeutic regimen of Curcumin (oral suspension of 150 mg Curcumin kg-1 rat weight). Sixteen adult male Wistar rats were assigned to four groups: Control, Group I (Curcumin only), Group II (Curcumin administration, then induced inflammation after 1 hour) and Group III (induced inflammation then Curcumin administration after 2 hours). Oxidative stress was assessed by measuring serum malondialdehide and carbonylated proteins, while systemic and local total antioxidant capacity was determined by ABTS. Local tissue changes (muscle, kidney, liver) were analysed using histopathology. Results showed that acute inflammation significantly increased lipid peroxidation in Groups II and III compared to Control and Group I. A significantly reduced total antioxidant capacity (ATBS) was present in serum and kidney in Group II, also in muscle and kidney in Group III. ABTS levels were significantly increased only in the liver tissue of the animals in Groups II and III with induced inflammation as compared to Group I. This study proved the potential of Curcumin in reducing oxidative stress in both prophylactic and therapeutic regimens.

scholarly journals Neuroprotective effects of Zingerone against carbon tetrachloride (CCl4) induced brain mitochondrial toxicity in Swiss albino mice

2018 ◽  
Vol 10 (2) ◽  
pp. 548-552
Author(s):  
Mohammad Firoz Alam
Keyword(s):  
Oxidative Stress ◽  
Carbon Tetrachloride ◽  
Antioxidant Enzyme ◽  
Brain Mitochondria ◽  
Control Group ◽  
Mitochondrial Toxicity ◽  
Swiss Albino Mice ◽  
Group I ◽  
Albino Mice ◽  
Group Ii

The present study targeted the brain mitochondria dysfunction in Swiss albino mice through carbon tetrachloride intoxication and its treatment with Zingerone. It is proposed that brain mitochondria is the main organelle responsible for oxidative stress by producing  reactive oxygen species (ROS). Swiss albino mice were divided into four groups; Group-1 was control; Group-2 was carbon tetrachloride (CCl4) toxic (1.5mg kg-1 bm i.p two days in a week.); Group-3 was pretreated with Zingerone (100 mg kg-1 b.m)  a day before  the administration of CCl4 and Group-4 was only Zingerone (100 mg kg-1 bm) given orally for 15days once in a day. At the end of the experiment mice were sacrificed and mitochondria were isolated from brain. Isolated brain mitochondria were further analyzed for oxidative stress marker. Thiobarbituric acid reactive substance (TBARS) content was increased significantly by CCl4 administration in Group-II as compared to the control Group-I, while the antioxidant (GSH) and other antioxidant enzyme GPx , GR, and CAT was depleted significantly in CCl4 treated Group-II as compare to control Group-I. Zingerone protected the  toxicity of brain mitochondria by reducing the lipid peroxidation and enhancing the antioxidant enzyme in Group-III and there was no significant changes were noticed in Group-IV as  compared to Group-I. Overall results showed the potential effects of Zingerone in protecting the neuronal cell loss by oxidative stress. Thus, the  present study indicated that the Zingerone may be used as the potential therapeutic tools for the prevention of CCl4 induced brain mitochondrial toxicity.  

scholarly journals The Effect of Intratesticular Injection of Human Adipose-Derived Stem Cell (hADSC) on Testicular Oxidative Stress and Spermatogenesis Process in the Varicocele Rat Model

2021 ◽  
Author(s):  
Safendra Siregar ◽  
Bambang Sasongko Noegroho ◽  
Ricky Adriansjah ◽  
Akhmad Mustafa ◽  
Ananta Bonar
Keyword(s):  
Oxidative Stress ◽  
Rat Model ◽  
Wistar Rats ◽  
Histopathological Examination ◽  
Therapy Group ◽  
Intervention Group ◽  
Testicular Tissue ◽  
Control Group ◽  
Group I ◽  
Group Ii

Abstract Introduction: Varicocele is the predominant cause of male infertility and was found in 19% - 41% of men with primary infertility and 45% - 81% of men with secondary infertility. Human adipose Derived Stem Cells (hADSC) can suppress oxidative stress in some oxidative injury model. Therefore, this study would like to investigate the effect of intratesticular hADSC injection on MDA level and spermatogenesis process by histopathological examination in the varicocele rat model.Method: This is an experimental study. A total sampling of 9 male Wistar rats were divided into three groups. Group I consist of 1 Wistar rats without any treatment or model (sham group), group II consist of 4 Wistar rats with varicocele model without hADSC therapy (control group), and group III consist of 4 Wistar rats with varicocele model and were given injections of 1.0x106 hADSC cells intratesticularly 30 days after model was made (therapy group). Testicular tissue was harvested for evaluation. Results: In all varicocele model rats (group II and III), the result of MDA level in therapy group (2.53 mol/liter) was significantly lower than the MDA level in control group (4.43 mol/liter) (p = 0.01). On histopathological examination, the average Johnson's Score in the therapy and control group was 9,77 and 9,18, respectively. The analysis showed Johnson’s score in the intervention group was significantly higher (p = 0.018). Conclusion: Intratesticular injection of hADSC can help reduce MDA levels and improve spermatogenesis process, which is damaged by varicoceles.

Comparison of Extenders for Liquid Storage of Indian Red Jungle Fowl (Gallus Gallus Murghi) Spermatozoa

2016 ◽  
Vol 9 (3) ◽  
pp. 207-212 ◽  
Author(s):  
Bushra Allah Rakha ◽  
Muhammad Sajjad Ansari ◽  
Iftikhar Hussain ◽  
Maqsood Anwar ◽  
Shamim Akhter ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Semen Quality ◽  
Membrane Integrity ◽  
Storage Period ◽  
Gallus Gallus ◽  
Quality Parameters ◽  
Liquid Storage ◽  
Red Jungle Fowl ◽  
Semen Extender ◽  
Acrosomal Integrity

This study was designed to evaluate a range of avian semen extenders for liquid storage of Indian Red Jungle Fowl ( Gallus gallus murghi) spermatozoa at 5 °C. Semen was collected from 8 mature trained cocks and processed in the Beltsville Poultry, Turkey, Lake, EK, Tselutin Poultry and Chicken semen extenders for storage at 5 °C. Semen quality parameters viz, motility (%), plasma membrane integrity (%), livability (%) and acrosomal integrity (%) were assessed at 0, 3, 6, 24 and 48 hours of storage. A time dependent decrease was observed in motility, plasma membrane and acrosomal integrity in all experimental extenders during the storage period. However, the Turkey semen extender was found significantly ( P<0.05) superior for protecting all aforementioned semen quality parameters compared to the Beltsville Poultry, Lake, EK, Tselutin Poultry and Chicken semen extenders. It is concluded that the Turkey semen extender can be used efficiently for the liquid storage of Indian Red Jungle Fowl spermatozoa at 5 °C.

scholarly journals Butaphosphan and Cyanocobalamin Supplementation in Semen Extender on Chilled Boar Sperm Quality and Life Span

2020 ◽  
Vol 7 ◽  
Author(s):  
J. Suwimonteerabutr ◽  
S. Chumsri ◽  
P. Tummaruk ◽  
Morakot Nuntapaitoon
Keyword(s):  
Plasma Membrane ◽  
Life Span ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Control Group ◽  
Plasma Membrane Integrity ◽  
Progressive Motility ◽  
Boar Sperm ◽  
Acrosome Integrity ◽  
Semen Extender

The objective of the present study was to determine the effect of butaphosphan and cyanocobalamin supplementation in semen extender on chilled boar sperm quality and life span. A total of 35 ejaculates of boar semen were included. The semen was diluted with Beltsville thawing solution extender supplemented with different concentrations of butaphosphan and cyanocobalamin [0 (control), 0.1, 0.2, 0.3, 0.4, and 0.5%] in the diluted semen. The semen samples were evaluated using a computer-assisted sperm analysis system to determine sperm motility and sperm kinetic parameters (i.e., the curvilinear velocity, VCL; straight line velocity, VSL; average path velocity, VAP; linearity, LIN; straightness, STR; amplitude of lateral head, ALH; wobble, WOB; and beat cross frequency, BCF). Additionally, sperm viability, acrosome integrity, mitochondrial activity, and plasma membrane integrity were evaluated after 4 (day 0), 72 (day 3), 120 (day 5), and 168 (day 7) h of storage using SYBR-14–ethidium homodimer-1 (EthD-1), EthD-1, JC-1, and the short hypo-osmotic swelling test, respectively. The analyses were carried out by using the general linear mixed model (MIXED) procedure of SAS. The statistical models for each data set included group, day after storage, and interaction between group and day after storage. The boar was included as a random effect. On day 0 after storage, progressive motility, VCL, VSL, VAP, and plasma membrane integrity of boar sperm in 0.3% of butaphosphan and cyanocobalamin supplementation were greater than those in the 0.4 and 0.5% groups (P &lt; 0.05). On day 3 after storage, total motility and progressive motility, VCL, VSL, VAP, LIN, WOB, BCF, and plasma membrane integrity in 0.3% of butaphosphan and cyanocobalamin supplementation were significantly greater than those in the control group (P &lt; 0.05). The total motility and progressive motility, VAP, and WOB in 0.3% of butaphosphan and cyanocobalamin supplementation were greater than those in the control group on day 5 after storage (P &lt; 0.05). No effects of butaphosphan and cyanocobalamin supplementation on acrosome integrity and mitochondria activity were found on days 3, 5, and 7 after storage. However, the motility and progressive motility and the values for all sperm kinetic parameters except ALH in 0.3% of butaphosphan and cyanocobalamin supplementation were greater than those in the control group on day 7 after storage (P &lt; 0.05). In conclusion, 0.3% of butaphosphan and cyanocobalamin supplementation in semen extender improved sperm motility, sperm activity, morphology, and life span in chilled boar sperm.

Evaluation and correlation of oxidative stress and haemato-biochemical observations in horses with natural patent and latent trypanosomosis in Punjab state of India

2018 ◽  
Vol 63 (4) ◽  
pp. 733-743 ◽  
Author(s):  
Rahul Parashar ◽  
Lachhman Das Singla ◽  
Mayank Gupta ◽  
Suresh Kumar Sharma
Keyword(s):  
Oxidative Stress ◽  
Negative Control Group ◽  
Negative Control ◽  
Control Group ◽  
Group Iii ◽  
Group I ◽  
Group Ii ◽  
Latent Group ◽  
Biochemical Indices ◽  
And Oxidative Stress

Abstract The haemato-biochemical indices and oxidative stress markers in horses naturally infected with Trypanosoma evansi were evaluated by analyzing the level of these parameters between T. evansi infected (microscopically positive patent group and PCR positive latent group) and infection free horses. To compare the hemato-biochemical indices and oxidative stress indicators, horses were divided into three categories based on diagnostic test employed and positive results obtained. These included Romanowsky stained slide positive group (Group I; n = 6), PCR positive group (group II; n = 28) and negative control group (group III, n = 30), revealing parasitologically positive patent, molecular positive latent and disease free status of horses. A significant reductions in total erythrocytes count (TEC, P = 0.01), haemoglobin (Hb, P = 0.01) and packed cell volume (PCV, P = 0.04) was noticed both in group I and group II while significant neutrophilia and lymphocytopenia was observed in group I when compared to negative control group. Substantial increase in creatinine (CRTN, P = 0.032) and gamma glutamyl transferase (GGT, P = 0.012) in group I while significant decrease in glucose (GLU, P = 0.04) and iron (Fe, P = 0.01) were noticed in both group I and group II in comparison to group III. A significant difference in lipid peroxides (LPO, P = 0.01) with highest level in patent group I (15.33 ± 0.53) followed by PCR positive latent group (14.09 ± 1.66) indicates higher lipid peroxidation in erythrocytes and oxidative stress in decreasing order when compared with infection free control horses (9.83 ± 0.97). Catalase (CAT, P = 0.01) was significantly lower in parasitological (0.82 ± 0.14) and molecular positive cases (1.27 ± 0.35) in comparison to control group (3.43 ± 0.96). The levels of superoxide dismutase (SOD, P = 0.01), reduced glutathione (GSH, P = 0.01) and ferric reducing antioxidant power (FRAP, P = 0.01) were significantly lower in parasito-molecular positive cases as compared to infection free control horses. An inverse correlation of RBC count with LPO and GSH and a direct correlation with catalase, SOD and FRAP was revealed. Overall, the observed substantial decreases in the oxidative parameters like catalase CAT, SOD, GSH and FRAP activities with remarkably elevated levels of LPO indicate high exposure of erythrocytes to oxidative damage in T.evansi infected horses.

Study on Keeping Quality and Cryopreservability of Buffalo Semen in Egg Yolk and Soybean Based Extenders

2017 ◽  
Vol 12 (3) ◽  
Author(s):  
D. V. Chaudhari ◽  
A. J. Dhami ◽  
K. K. Hadiya ◽  
J. A. Patel
Keyword(s):  
Egg Yolk ◽  
Progressive Motility ◽  
Keeping Quality ◽  
Buffalo Semen ◽  
Acrosome Integrity ◽  
Semen Extender ◽  
Refrigeration Storage ◽  
Surti Buffalo ◽  
Fresh Semen ◽  
Acrosomal Integrity

The study was aimed to compare efficacy of egg yolk based diluent (TFYG) with soybean based commercial diluents (Bioxcell® and Optixcell®, IMV, France) for refrigeration and cryopreservation (-196°C) of six Surti buffalo semen. Each qualified ejaculate (n=8/bull, >70% initial motility) was splitdiluted @ 100 ×106 sperm ml-1 at 34°C with 3 diluents. Part of each aliquot was filled in French mini straws and rest transferred to refrigerator for gradual cooling to 4-5°C. After 4 h of equilibration at 4-5°C in cold handling cabinet, the straws were frozen using programmable bio-freezer. The initial mean motility, acrosome integrity and hypo-osmotic reactive sperm per cent of fresh semen were 78.54±0.30, 94.40±0.20 and 79.35±0.42%, respectively. Sperm progressive motility at 24 hrs of refrigeration storage was 68.33±0.45, 66.25±0.46 and 70.31±0.46 %, and at 72 hrs of refrigeration storage 52.50±0.68, 48.54±0.84 and 53.85±0.75 % in TFYG, Bioxcell and Optixcell diluents, respectively. Acrosomal integrity and hypo-osmotic reactivity at 24 hrs of refrigeration storage were 88.15±0.18, 87.25±0.21, 89.27±0.20% and 66.04±0.50, 63.81±0.45, 68.10±0.46%, whereas at 72 hrs of refrigeration, the values were 81.67±0.23, 80.60±0.30, 82.83±0.27% and 51.35±0.60, 47.35±0.68, 53.40±0.68%, respectively, in above 3 diluents. The pre-freeze sperm progressive motility recorded in TFYG, Bioxcell and Optixcell diluents was 69.48±0.37, 68.02±0.49 and 70.94±0.38%, and post-thaw motility was 47.71±0.79, 44.38±0.85 and 49.90±0.90%, respectively. Per cent sperm acrosomal integrity and hypo-osmotic reactivity in TFYG, Bioxcell and Optixcell diluents were 89.54±0.18, 88.58±0.22, 90.52±0.21% and 67.96±0.32, 65.65±0.42, 70.23±0.37% at pre-freeze stage, whereas 76.83±0.23, 75.90 ±0.27, 78.50±0.25% and 45.02±0.84, 42.31±0.82, 47.81±0.90% at post-thaw stage, respectively. The post-thaw longevity after 60 minutes of incubation (37°C) was 35.52±0.79, 31.15±0.85, 37.19±0.81%, respectively, in above 3 diluents. Among the three semen diluent percent sperm progressive motility, acrosomal integrity and hypo-osmotic reactivity at particular stage were higher in Optixcell showing at par results in TFYG, whereas Bioxcell showed significantly (P less than 0.05) lower findings than other two diluents. Optixcell is a commercial product much costlier than TFYG, hence there is a need to develop local soybean based semen extender which can give comparable results with TFYG.

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