Cryopreservation of Indian Red Jungle Fowl (Gallus Gallus Murghi) Spermatozoa: A First Approach

2017 ◽  
Vol 10 (1) ◽  
pp. 6-11 ◽  
Author(s):  
Bushra Allah Rakha ◽  
Muhammad Sajjad Ansari ◽  
Iftikhar Hussain ◽  
Maqsood Anwar ◽  
Shamim Akhter ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Gallus Gallus ◽  
Freeze Thaw ◽  
Red Jungle Fowl ◽  
Frozen Semen ◽  
Sperm Plasma Membrane ◽  
Acrosomal Integrity

The experiment was designed to show the capacity of Indian Red Jungle Fowl ( Gallus gallus murghi) sperm to be cryopreserved. The changes in characteristics of Indian Red Jungle Fowl spermatozoa during different phases of cryopreservation with the glycerol cryoprotectant and straw packaging were recorded. Semen from eight mature cocks were pooled, diluted at 37 °C in a Tselutin poultry extender, cooled to 4 °C in 2 h (0.275 min−1) and equilibrated for 10 min after the addition of 11% glycerol. Cooled semen was filled in 0.5 mL French straws, kept over liquid nitrogen vapour for 10 min and plunged into liquid nitrogen. Frozen semen was thawed at 37 °C for 30 s. Sperm motility, plasma membrane plasticity, livability, and acrosomal integrity were assessed at the stage of pre-dilution (37 °C), post-dilution, cooling, equilibration and freeze-thawing. The experiment was repeated five times. Sperm motility did not change at post-dilution and cooling and averaged 83.3±3.1%. However, motility declined ( P<0.05) to 66.0±2.4% and 45.0±6.5% at post-equilibration and thawing stages, respectively. The sperm plasma membrane plasticity at the pre-dilution stage was 74.8±1.4% and decreased ( P<0.05) to 62.3±1.7%, 51.7±1.7%, and 42.0±1.2% at the stages of post-dilution, cooling and freeze-thawing, respectively. A decline ( P < 0.05) in acrosomal integrity was observed at post-cooling (83.8±2.4%), equilibration (80.8±0.8%) and thawing (73.8±2.4%). It is concluded that Indian Red Jungle sperm seem to adapt quite well to the freeze-thaw process that mainly alters the motility capacities and much less the acrosomal integrity. To our knowledge, this is the first ever report on cryopreservation of Indian Red Jungle Fowl spermatozoa.

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.

Cryoprotective Effect of Glycerol Concentrations on Indian Red Jungle Fowl (Gallus Gallus Murghi) Spermatozoa

2018 ◽  
Vol 11 (2) ◽  
pp. 80-88 ◽  
Author(s):  
Bushra Allah Rakha ◽  
Muhammad Sajjad Ansari ◽  
Shamim Akhter ◽  
Elisabeth Blesbois
Keyword(s):  
Plasma Membrane ◽  
Semen Quality ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Gallus Gallus ◽  
Recovery Rates ◽  
Plasma Membrane Integrity ◽  
Red Jungle Fowl ◽  
Frozen Semen ◽  
Acrosome Integrity

Semen cryopreservation protocols for wild avian species need to be optimised in order to achieve optimum post-thaw sperm quality and fertility. The present study was designed to evaluate the cryoprotective effect of different glycerol concentrations (11%, 15% and 20%) on post-thaw quality, recovery rates, absolute livability index and fertility of Indian Red Jungle Fowl (Gallus gallus murghi) semen. Semen was collected from eight mature cocks and cryopreserved for storage at −196 °C. Frozen semen was thawed at 37 °C for 30 s and assessed for motility, plasma membrane integrity, viability and acrosome integrity at 0, 2 and 4 h incubation at 37 °C. Percentages of motility, plasma membrane integrity, viability and acrosome integrity were recorded higher (P<0.05) post-thaw at 0, 2 and 4 h at 37 °C with 20% glycerol compared to 15% and 11% glycerol. Likewise, recovery rates (%) of aforementioned parameters after cryopreservation and absolute livability index were observed highest (P<0.05) with 20% glycerol. By comparing values of R2 after multivariate regression analysis, least negative effects of hours of incubation were observed on semen quality in extenders with 20% glycerol followed by 15% and 11% glycerol. The fertility outcomes (number of fertile eggs, fertility [%], number of hatched chicks, percent hatch and hatchability of fertilised eggs) were recorded higher (P<0.05) with 20% glycerol followed by 15% and 11% glycerol. It is concluded that the concentration of 20% glycerol gives the best cryoprotection for quality and fertility of Indian Red Jungle Fowl semen.

scholarly journals Fertilitas Semen Beku dalam Tris Kuning Telur dan Skim yang Diberi Omega-3 pada Sapi Simmental dengan Ransum Berimbuhan Seng dan Selenium Minimal

2018 ◽  
Vol 19 (2) ◽  
pp. 251 ◽  
Author(s):  
Asep Kurnia ◽  
Soeparna Soeparna ◽  
Raden Iis Arifiantini ◽  
Rahmat Hidayat
Keyword(s):  
Plasma Membrane ◽  
Liquid Nitrogen ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
The Other ◽  
Omega 3 ◽  
Nitrogen Vapor ◽  
Frozen Semen ◽  
Sperm Plasma Membrane ◽  
The Individual

This study aims to examine the quality of frozen semen in Tris egg yolk (TEY) extender and skimmed milk extender with or witout omega-3. A total of 18 Simmental bulls belong to Lembang Artificial Insemination Centre were divided into three groups. Each was fed with standard feed (R1), standard feeds supplemented with minimal Se and Zn (R2) and standard feed supplemented with maximal of Se and Zn concentration. Semen were collected using an artificial vagina and were evaluated macro- and microscopically. The semen then were divided into four tubes and each diluted with Skimmed, SkimmedOmega 3, TEY or TEY-O. The semen was then packed into a 0.25 ml straw and equilibrated at 5 oC for 4 hours, then frozen above liquid nitrogen vapor, and stored in liquid nitrogen container (-196 oC). The qualities of frozen semen were evaluated on the motility, individual score, viability and integrity of the plasma membrane of sperms. Sperm motility of bulls fed with standard feed (R1) in TEY extender and R3 in TEY and TEY-Omega-3 extender were higher (p <0.05) compared to the other combinations. No difference was found on the individual score. The viability of sperms in bulls fed with standard feed in SkimmedOmega-3 extender was higher than the other treatments and the highest sperm plasma membrane integrity was demonstrated by sperm in bull feeding with R2 in TEY extender.

71 BOVINE SPERM DEATH KINETICS: CHANGES IN MOTILITY, ACROSOMES, AND PLASMA MEMBRANE

2013 ◽  
Vol 25 (1) ◽  
pp. 183
Author(s):  
M. Ahmad ◽  
N. Ahmad ◽  
A. Riaz ◽  
M. Anzar
Keyword(s):  
Plasma Membrane ◽  
Statistical Analysis ◽  
Sperm Motility ◽  
Propidium Iodide ◽  
Female Reproductive Tract ◽  
Semen Sample ◽  
Membrane Asymmetry ◽  
Progressive Motility ◽  
Bovine Sperm ◽  
Acrosomal Integrity

Extent and timing of alterations in structures and functions of sperm after its placement in the female reproductive tract are important for successful fertilization. To our knowledge, the few reports are available on the kinetics of alterations in bovine sperm structures and functions during pathway to their death. Therefore, the present study was conducted to determine the changes in motility, acrosome and plasma membrane asymmetry in fresh and frozen–thawed semen during incubation at 37°C over the period of 24 h. Semen was collected from 3 breeding beef bulls, pooled, and considered as one replicate (total replicates = 5). Each pooled semen sample was diluted in Tris-citric acid egg yolk glycerol extender (pH 6.8), cooled to +4°C over 90 min, and then cryopreserved by a programmable cell freezer. Fresh (pooled semen) and frozen–thawed semen were incubated at 37°C for 24 h. Each semen sample was evaluated for sperm motility with computer-assisted semen analysis and acrosomal integrity and plasma membrane asymmetry using fluorescein isothiocyanate-peanut agglutinin/propidium iodide and Annexin V/propidium iodide assays, respectively, at 0, 2, 4, 6, 12, and 24 h of incubation at 37°C, with a flow cytometer. Statistical analysis was conducted using PROC MIXED model in statistical analysis system as 2 (semen types) × 6 (times) factorial model, using time as repeated measure. Progressive motility was higher (P < 0.05) in fresh than in frozen–thawed semen until 6 h. Progressive motility declined (P < 0.05) below the threshold level (i.e. 30%) much later (12 h) in fresh as compared with frozen–thawed semen (2 h). However, acrosomal integrity and plasma membrane asymmetry deteriorated (P < 0.05) below threshold at the same time interval (2 h) in both fresh and frozen–thawed semen. Viable sperm (AN–/PI–) remained higher (P < 0.05) during the first 6 h in fresh than in frozen–thawed semen and declined (P < 0.05) below the threshold at 12 h in fresh and at 6 h in frozen–thawed semen. In fresh semen, the necrotic sperm (AN–/PI+) population increased (P < 0.05) over time and reached maximum (97%) at 24 h. In frozen–thawed semen, a mixed population of late apoptotic (53%) and necrotic (34%) sperm was found at 24 h. In conclusion, the alterations in sperm motility, acrosomes, plasma membrane integrity, and asymmetry were slower in fresh than in frozen–thawed semen. Fresh sperm followed necrosis and frozen–thawed sperm underwent necrosis and apoptosis-like pathways, respectively. This study was supported by the Canadian Commonwealth Scholarship Program by the Canadian Bureau for International Education (CBIE), and Agriculture and Agri-Food Canada.

187 EXPLOITATION OF IN VITRO CAPACITATION FOR NANOPARTICLE INCORPORATION WITHIN MAMMALIAN SPERMATOZOA

2016 ◽  
Vol 28 (2) ◽  
pp. 224
Author(s):  
L. Myles ◽  
C. Durfey ◽  
P. Ryan ◽  
S. Willard ◽  
J. Feugang
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Surface Membrane ◽  
Milk Powder ◽  
Noninvasive Evaluation ◽  
Control Group ◽  
Body Tissues ◽  
Sperm Plasma Membrane

Migration and interactions of mammalian gametes occur in deep body tissues after mating, rendering difficult any in situ noninvasive evaluation of their performances with current methods. In our effort to develop an effective and real-time in vivo imaging approach, we have successfully labelled porcine gametes with self-illuminating bioluminescent and red-shifted quantum dot nanoparticles (QD) in our previous studies (Feugang et al. 2012 J. Nanobiotechnol. 10, 45; Feugang et al. 2015, J. Nanobiotechnol. 13, 38). The present effort aimed at investigating whether QD could be incorporated into spermatozoa through induced in vitro capacitation, which increases sperm plasma membrane fluidity. Fresh extended boar semen was placed on top of a Percoll gradient and centrifuged. Purified motile spermatozoa were collected and washed with pre-warmed PBS. Pelleted spermatozoa were resuspended in the modified Tris-buffered medium with BSA fraction-V (1 mg mL–1; modified Tween medium B with milk powder and BSA). Sperm aliquots (108) were supplemented or not (control) with QD only (QD+; 1 nM), QD+caffeine (2 mM), or QD+heparin (10 µg mL–1); with caffeine and heparin being used as routine capacitant agents in fertilization media. All aliquots were incubated at 38.5°C, under 5% CO2 for 0.5, 1, or 3 h. Spermatozoa were then analysed for motility characteristics and imaged for confirmation of QD-sperm interactions (bioluminescence emission) and localization (transmission electron microscope; TEM). Motility data of 5 replicates were analysed with ANOVA-2, and P < 0.05 was set as threshold of significance. Total sperm motility (TSM) significantly improved with the presence of either or both QDs and capacitant agents after 0.5 and 1 h incubations. With exception of the QD+heparin, all other groups had significantly decreased TSM after 3 h of incubation, when compared with TSM at 0.5 and 1 h. Higher proportions of progressive and rapid (≥45 µm s–1) spermatozoa were observed in the presence of both capacitant agents (P < 0.05), and only QD+heparin maintained greater proportions after 3 h. Sperm straight-line velocity significantly increased in the QD+caffeine at 0.5 h and in both QD+caffeine and QD+heparin thereafter. Sperm straightness data were increased by both caffeine and heparin during incubations. Strong bioluminescence signals were observed in spermatozoa incubated with QDs compared to the background signal seen in the control group. The TEM images revealed consistent surface membrane attachment of QDs in all QD+ groups, whereas transmembrane and intra-spermatic localizations were visible in both QD+caffeine and QD+heparin groups. We concluded that supplementations of medium containing QDs with caffeine or heparin allow the crossing of sperm plasma membrane by QD. No toxic effect of QD on sperm motility was observed, which confirmed our previous report using a similar ratio of QDs over spermatozoa. Exploration of efficient incorporation of QD into spermatozoa as a promising approach for noninvasive molecular imaging is still ongoing, as well as further sperm viability assessments. Supported by the NIH grant #5T35OD010432 and USDA-ARS Biophotonics Initiative grant #58–6402–3-0120.

scholarly journals ESTUDO PRELIMINAR DE ALGUMAS CARACTERÍSTICAS DO SÊMEN CANINO CONGELADO

2000 ◽  
Vol 5 (1) ◽  
Author(s):  
R. R. WEISS ◽  
S. RODASKI ◽  
J. M. BÜCHELE ◽  
I. W. SANTOS ◽  
L. M. ALMEIDA
Keyword(s):  
Liquid Nitrogen ◽  
Final Concentration ◽  
Spermatozoa Motility ◽  
Frozen Semen ◽  
Preliminary Study ◽  
Acrosomal Integrity

Na crioconservação de sêmen canino, técnicas de congelamento precisam ser otimizadas com a finalidade de maximizar a viabilidade do sêmen após a descongelação. O ejaculado de um animal foi congelado em palhetas de 0,5ml utilizando como diluidores o Tris-citrato-frutose, Tris-citrato-glucose, com 6% de glicerol no volume final, e o diluente Merk. As amostras foram avaliadas imediatamente após descongelação quanto à motilidade e integridade de acrossomas. As mais altas taxas de motilidade e integridade acrossomal foram obtidas com o diluente Tris-citratoglucose, havendo diferenças significativas quando comparadas com os demais diluentes, demonstrando a variação na viabilidade do sêmen de um mesmo animal frente a diferentes diluidores. Preliminary study on some properties of canine frozen semen Abstract Current techniques for freezing canine semen ought to be improved in order to attain maximum viability from ejaculates. In the present study, three aliquots of an ejaculate from one dog has been diluted in three different extenders: TRIS-citrate-fructose, TRIS-citrate-glucose and Merck extender. Glycerol in the final concentration of 6% has been added to TRIS-citrate-fructose and TRIS-citrate-glucose extenders after the semen dilution. After the dilution, all three preparations were cooled first at 5ºC and then frozen in liquid nitrogen at – 196ºC. Spermatozoa motility and acrosomal integrity were assessed immediately after thawing. The values for the rate of motility and the proportion of spermatozoa displaying intact acrosome found in the TRIS-citrate-glucose preparation were significantly superior when compared with the values found for TRIS-citrate-fructose and the Merk extender.

scholarly journals MIG-23 is involved in sperm migration in Ascaris suum

2021 ◽  
Author(s):  
Xia Wang ◽  
Qiushi Wang ◽  
Ruijun He ◽  
Qi Zhang ◽  
Jin Shan ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Atpase Activity ◽  
Sperm Motility ◽  
Ascaris Suum ◽  
Nucleoside Triphosphate ◽  
Mitochondrial Activity ◽  
Major Sperm Protein ◽  
Sperm Activation ◽  
Sperm Migration ◽  
Sperm Plasma Membrane

Sperm motility acquisition during maturation is essential for successful fertilization.Extracellular adenosine-5'-triphosphate (ATP) level mediation by MIG-23, which is a homolog of human ecto-nucleoside triphosphate diphosphohydrolase (E-NTPDase), was required for major sperm protein filament dynamics and sperm motility in the nematode Ascaris suum. MIG-23 was localized on the sperm plasma membrane. During sperm activation, mitochondrial activity was increased dramatically, and a large amount of ATP was produced and stored in refringent granules (RGs). In addition, a portion of the produced ATP was released to the extracellular space through ATP channels, which were composed of innexins and localized on the sperm plasma membrane. Spermatozoa, instead of spermatids, hydrolyzed exogenous ATP and processed ecto-ATPase activity. MIG-23 contributed to the ecto-ATPase activity of spermatozoa. MIG-23 activity was interrupted, spermatozoa also decreased their ATP hydrolysis activity. Blocking MIG-23 activity resulted in an increase in the depolymerization rate of MSP filaments in pseudopodia, which eventually affected nematode sperm migration. Overall, our data imply that MIG-23, which contributes to the ecto-ATPase activity of spermatozoa, regulates sperm migration by modulating extracellular ATP levels.

21 Effect of egg yolk extracted low-density lipoprotein on cryopreserved Nguni bull semen

2019 ◽  
Vol 31 (1) ◽  
pp. 136
Author(s):  
M. M. Tshabalala ◽  
K. A. Nephawe ◽  
M. L. Mphaphathi ◽  
C. M. Pilane ◽  
T. L. Nedambale
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Sperm Quality ◽  
Egg Yolk ◽  
Semen Parameters ◽  
Dna Integrity ◽  
Low Density ◽  
Bull Semen ◽  
Frozen Semen ◽  
Live Sperm

Egg yolk has been reported to have a beneficial effect on sperm quality following cryopreservation, and this led to its widespread use in semen extenders. However, egg yolk contains substances that inhibit respiration of sperm cells and diminish their motility rate. Moreover, it also contains low-density lipoproteins (LDL) that have a protective effect on sperm during the cryopreservation process. The objective of this study was improve cryopreservation of Nguni bull semen using egg yolk low-density protein. A total of 25 ejaculates were collected from 5 Nguni bulls aged 4 to 5 years using an electroejaculator during the natural breeding season. Collected raw semen samples were transported to the laboratory and evaluated for sperm motility, viability, plasma membrane, acrosome, and DNA integrity before dilution. Semen was randomly diluted with a sodium citrate extender supplemented with 20% egg yolk (control) and with 6, 8, 10, and 12% LDL concentrations. The diluted semen sample groups were equilibrated for 4h at 5°C. Following equilibration, semen was loaded into 0.25-mL straws and frozen in a controlled-rate programmable freezer. The groups of semen straws were then plunged into LN and transferred into LN tanks (−196°C) for storage. The frozen semen straws per treatment group were thawed at 37°C and evaluated for sperm motility, viability, plasma membrane, acrosome, and DNA integrity. Data were analysed with ANOVA using Stata V12 statistical software (StataCorp., College Station, TX), and treatment means were separated using Fisher’s protected t-test at the significant level of P&lt;0.05. Sperm motility and membrane integrity were significantly higher (P&lt;0.05) in frozen-thawed semen diluted with 8% LDL compared with the other concentrations. However, 6 and 8% LDL resulted in a significantly higher (P&lt;0.05) live sperm, DNA, and acrosome integrity. Frozen-thawed semen diluted with 10 and 12% LDL resulted in the lowest percentages of sperm motility, live sperm, plasma membrane, acrosome, and DNA integrity following cryopreservation. In conclusion, extender containing 8% LDL resulted in improved Nguni bull semen parameters such as sperm motility, viability, plasma membrane, acrosome, and DNA integrity following cryopreservation. Further studies are required to determine the fertilizing capacity of semen diluted and frozen with LDL in vitro and in vivo.

Semen quality and fertility of liquid stored and frozen-thawed semen in crossbred pigs of North-Eastern India

2018 ◽  
Author(s):  
G Kadirvel ◽  
M K Kalita ◽  
Raju Kr Dewry ◽  
Ashok Kumar ◽  
Nripendra Mahanta ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Sperm Motility ◽  
Semen Quality ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Eastern Region ◽  
Plasma Membrane Integrity ◽  
Frozen Semen ◽  
North Eastern ◽  
Farrowing Rate

Study was conducted to compare the semen quality and fertility of liquid stored semen for three days and frozen-thawed semen in the north-eastern region of India. For liquid semen, the semen ejaculates were extended in Beltsville Thawing Solution (BTS) extender and preserved at 17°C for three days. For cryopreservation, semen was diluted Lactose-egg yolk-glycerol extender and frozen in straw using programmable freezer with freezing rate of 40°C/min from -6 to -140°C. The preserved evaluated for sperm motility, viability, plasma membrane integrity and fertility. The results revealed that the liquid stored semen has maintained the sperm motility and viability up to day 3 without significant reduction. Similarly the plasma membrane integrity did not differ significantly up to day 2, but it was significantly (P less than 0.05) reduced on days 3 in liquid stored semen. After freezing and thawing, the mean sperm motility, viability and plasma membrane integrity were 58.25 ± 2.96%, 64.75 ± 2.47% and 47.06 ± 2.02%, respectively. These parameters were significantly (PP less than 0.01) lower as compared to the liquid stored semen from day 0 to day 3. After insemination with liquid semen, the farrowing rate was 77.7%, 80.76%, 73.07% and 69.8%, respectively from day 0, day1, day 2 and day 3. The pregnancy rate, farrowing rate and litter size did not differ significantly among different days of liquid storage. These parameters were significantly (PP less than 0.01) lower in frozen semen as compare to that of liquid stored semen. The study concluded that the liquid semen stored up to three days is more efficient than frozen-thawed semen in terms of preserving sperm quality and fertility.

scholarly journals The effect of freezing on cryosurvival of yak sperm

2018 ◽  
Vol 15 (2) ◽  
pp. 215-218
Author(s):  
S Deori
Keyword(s):  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Artificial Insemination ◽  
Sperm Concentration ◽  
Egg Yolk ◽  
Mean Values ◽  
Frozen Semen ◽  
Sperm Abnormality ◽  
Semen Characteristics ◽  
Live Sperm

A study was carried out to study the effect of freezing on cryosurvival of yak semen. Artificial insemination in yak is still in infancy. Semen cryopreservation and use of artificial insemination can be applied in yak husbandry for conservation and rapid multiplication of superior germplasm. Semen was collected from four adult yak bulls using artificial vagina method managed under uniform conditions. A total of 40 ejaculates comprising of 10 ejaculates each bull were collected following twice a week schedule and evaluated for fresh semen characteristics. The fresh yak semen characteristics viz. ejaculate volume (ml), mass activity (0-4), initial sperm motility (%), sperm concentration (x 106/ml), live sperm (%), sperm abnormality (%) and intact acrosome (%) were 3.10 ± 0.18, 3.53 ± 0.96, 83.89 ± 2.87, 1180.22 ± 42.32, 77.63 ± 4.23, 8.45 ± 3.33 and 93.61 ± 3.78 respectively. The ejaculates were diluted (1:10) with Tris extender consisting of 6.4 ml glycerol and 20 ml of fresh egg yolk. Straws were equilibrated at 5°C for 4 hours followed by exposure to liquid nitrogen vapour for 10 minutes and finally transferred to liquid nitrogen container for storage. The cryosurvival rate was studied after 7 days of storage in liquid nitrogen. The frozen semen was thawed in warm water (37°C) for 30 seconds for evaluation. Mean values of postthaw sperm motility (%), live sperm (%) and intact acrosome (%) in yaks were 55.67 ± 4.67, 65.62 ± 3.23 and 89.26 ± 3.67 respectively. In conclusion, yak semen has a better cryosurvival while freezing in tris extender with 6.4 per cent glycerol and 20 per cent egg yolk following an equilibration period of 4h.SAARC J. Agri., 15(2): 215-218 (2017)

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