Effect of different levels of egg-yolk on freezability of Jakhrana buck semen

Author(s):  
Narendra Kumar ◽  
B. Rai ◽  
Chetna Gangwar ◽  
S. A. Lone ◽  
Anshuman Kumar ◽  
...  

The present study was designed to determine the effect of different levels of egg-yolk on freezability of Jakhrana buck semen. Six healthy Jakhrana bucks (BW=30 ± 2kg, age=12 ± 0.5 month) were used for semen collection. These bucks were maintained under semi-intensive system at Jakhrana Unit of C.I.R.G. Makhdoom, Mathura. A total of 48 ejaculates (6 bucks × 8 replicates) were collected twice a week using artificial vagina. Each ejaculate was divided into 4 groups (G1, G2, G3 and G4). The G1, G2, G3 and G4 were extended with Tris-egg yolk-citric acid- fructose-glycerol (TEYCFG) extenders containing 5, 10, 15 and 20% egg yolk level, respectively. Each ejaculate was evaluated for sperm motility, viability, abnormality, and hypo-osmotic swelling (HOS) response and acrosome integrity before and after freezing. At pre-freeze stage no significant (P>0.05) difference in sperm motility and viability was found among all groups. Sperm abnormality was significantly (P<0.05) higher in G4 as compared to other groups (G1, G2, G3). The HOS response and acrosomal integrity was significantly (P<0.05) higher in G1, G2 and G3 as compared to G4. However, no significant (P>0.05) difference was observed in HOS response and acrosomal integrity among G1, G2 and G3. At post thaw stage, sperm motility, viability and HOS response was significantly (P<0.05) higher in G1 and G2 as compared to G3 and G4. Sperm abnormality was significantly (P<0.05) lower in G2 as compared to other groups. The acrosomal integrity was significantly (P<0.01) higher in G1 and G2 as compared to G3 and G4. It is concluded that 10% egg yolk in Tris based extender may be the best for successful cryopreservation of Jakhrana buck semen.

2006 ◽  
Vol 18 (2) ◽  
pp. 160 ◽  
Author(s):  
S. Mosaferi ◽  
A. Niasari-Naslaji ◽  
N. Bahmani ◽  
A. A. Gharahdaghi ◽  
A. Abarghani ◽  
...  

Disaccharides have been used as an extender for dromedary camel semen (Bravo et al. 2000 Anim. Reprod. Sci. 62, 173-193). More recently we have investigated the effect of different concentrations of lactose extender on the viability of Bactrian camel spermatozoa (Mosafer et al. 2005 Reprod. Fertil. Dev. 17, 197). Considering the osmolality (316.1 � 1.48 mOsm/kg) and pH (7.4 � 0.03) of Bactrian camel semen (Mosaferi et al. 2005 Theriogenology 63, 92-101), the objective of this study was to investigate the effect of osmolality of sucrose extender on the viability of Bactrian camel spermatozoa. Sucrose at the concentrations of 9, 10, 11, 12, and 13% with osmolalities of 292, 331, 356, 386, and 410 mOsm/kg, respectively, were prepared. All extenders contained 20% egg yolk and antibiotics, with pH adjusted to 6.9. Semen was collected from camels with a sound history of semen quality and fertility (n = 3) using a modified artificial vagina and divided into different treatments after mechanical reduction of semen viscosity (3). Progressive forward motility of spermatozoa was examined at the time of semen collection and at 4, 12, and 24 h after incubation at 4�C. Data were analyzed using the GLM procedure in SAS/STAT after arcsin transformation. At the time of semen dilution, the progressive forward motility of spermatozoa was greater at osmolality of 331 (23%) compared with 292 (1%), 386 (6%), and 410 (3.5%) mOsm/kg (P < 0.05). No progressive forward motility of spermatozoa was noticed after 4 h incubation at 4�C at osmolalities of 292, 386, and 410 mOsm/kg. At this time, a significant decrease (P < 0.05) of progressive forward motility occurred at osmolalities of 331 (4%) and 356 (0.5%) compared with that of the time of dilution. After 12 and 24 h incubation at 4�C, no progressive forward motility of spermatozoa was detected in any of these extenders. In conclusion, 10% sucrose (331 mOsm/kg) at the adjusted pH of 6.9 was the most suitable concentration of this disaccharide for preserving Bactrian camel semen for less than 4 h under chilled conditions.


2005 ◽  
Vol 17 (2) ◽  
pp. 197 ◽  
Author(s):  
S. Mosaferi ◽  
A. Niasari-Naslaji ◽  
A.A. Gharahdaghi ◽  
A. Abarghani ◽  
A. Ghanbari ◽  
...  

Lactose has been used widely as a semen extender for camels although in the absence of evidence illustrating its suitablility. Considering the osmolarity (316.1 ± 1.48 mOsm/kg) and pH (7.4 ± 0.03) of Bactrian camel semen (Mosaferi S et al. 2004 Theriogenology, in press), the objective of this study was to investigate the effect of osmolarity and pH of lactose extender on the viability of Bactrian camel spermatozoa. In Experiment I, with pH adjusted to 6.9, the effect of lactose concentrations of 9, 10, 11, 12, and 13% with an osmolarity of 290, 333, 350, 376, and 419 mOsm/kg, respectively, on the viability of spermatozoa was investigated. In Experiment II, with lactose fixed at 10%, the effect of extender with pH of 5.9, 6.9, 7.5, 7.9, and 8.9 on the viability of spermatozoa was examined. All extenders contained 20% egg yolk. In both experiments, semen was collected from camels with a sound history of semen quality and fertility (n = 3), using a modified artificial vagina, and divided into different treatments after mechanical removal of semen viscosity (Mosaferi et al. 2004). Progressive forward motility of spermatozoa was examined at the time of semen collection and at 4, 12, and 24 h after incubation at 4°C. Data were analyzed using the GLM procedure in SAS/STAT (SAS Institute, Inc., Cary, NC, USA) after arcsine transformation. At the time of semen dilution, the progressive forward motility of spermatozoa was greater at osmolarity of 290 (28.5%), 333 (34%), and 350 (31%) compared to 376 (13.5%) and 419 (1%) mOsm/kg (P < 0.05). The same trend in the progressive forward motility of spermatozoa was noticed after 4 h of incubation at 4°C; although a significant decrease (P < 0.05) occurred at 290 (11%), 333 (18 %) and 350 (16%). After 12 and 24 h of incubation at 4°C, the progressive forward motility of spermatozoa was less than 10% at 333 and 350 mOsm/kg (P < 0.05). At the time of semen dilution, the progressive forward motility of spermatozoa was greater (P < 0.05) at pH 6.9 (35.5%) and 7.5 (18%) compared to pH 5.9 (0%), 7.9 (7.5%) and 8.9 (2.5%). The same trend in the progressive forward motility of spermatozoa was observed after 4 h incubation at 4°C; although, a significant decrease (P < 0.05) occurred at pH 6.9 (15%) and 7.5 (9%) at this time. After 12 h incubation at 4°C, the progressive forward motility of spermatozoa was less than 5% at pH 6.9 and 7.5 (P < 0.05). In conclusion, 10% (333 mOsm/kg) and 11% (350 mOsm/kg) lactose, at the adjusted pH of 6.9, were the most suitable concentrations of lactose extender for preserving Bactrian camel semen for less than 4 h after which the viability of spermatozoa deteriorated significantly in this extender. The authors wish to thank the director and station staff of Bactrian Camel Research Center at Jahadabad, Meshkinshahr, Ardabil, for kind provision of facilities and assistance throughout the experiment.


Reproduction ◽  
2004 ◽  
Vol 127 (5) ◽  
pp. 547-556 ◽  
Author(s):  
R E Spindler ◽  
Y Huang ◽  
J G Howard ◽  
P Wang ◽  
H Zhang ◽  
...  

Sperm cryopreservation and artificial insemination are important management tools for giant panda breeding and the preservation of extant genetic diversity. This study examined the influence of freeze–thawing on sperm function, specifically capacitation. Sperm from nine giant pandas were assessed before and after rapid (− 40 and − 100 °C/min) cryopreservation by incubation in HEPES-buffered Ham’s F10 medium with and without the capacitation accelerators, 3-isobutyl-1-methylxanthine (IBMX) and dibutyryl cyclic AMP (dbcAMP). At 0, 3 and 6 h of exposure, aliquots were assessed for sperm motility traits and capacitation, defined as the proportion of sperm with intact acrosomes following exposure to solubilised zonae pellucidae (ursid or felid) or calcium ionophore subtracted from the proportion of sperm with intact acrosomes before exposure. Although mean±s.e.m. sperm motility post-thaw (56.1 ± 3.9% at 0 h) was less (P < 0.05) than pre-freeze (71.7 ± 6.0%), there was no difference (P > 0.05) in the proportion of acrosome-intact sperm (fresh, 93.0 ± 1.7% versus cryopreserved–thawed, 81.7 ± 4.7% at 0 h). Incidence of capacitation was greater (P < 0.05) in fresh sperm incubated with capacitation accelerators IBMX and dbcAMP (9 h: 50.9 ± 1.1) compared with fresh sperm incubated without accelerators (9 h: 41.2 ± 1.1%). Frozen–thawed sperm preincubated without accelerators underwent capacitation (49.6 ± 1.1%) to a greater extent (P < 0.05) compared with these fresh counterparts. Thawed samples with (9 h: 45.9 ± 1.4%) and without accelerators (9 h: 41.2 ± 1.1%) did not differ (P > 0.05) during the 9-h incubation. We conclude that giant panda spermatozoa (1) undergo capacitation in vitro with or without chemical accelerators and (2) withstand a rapid cryopreservation protocol, including retaining normal acrosomal integrity and functional capacitation ability.


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.


SPERMOVA ◽  
2020 ◽  
Vol 10 (2) ◽  
pp. 74-80
Author(s):  
Wilber Garcia ◽  
◽  
Edwar Maxi ◽  
Veronica Macedo ◽  
Elizabeth Mendoza ◽  
...  

The objective was to evaluate the effect of two methods of freezing on seminal quality and effect of seminal plasma as an inducer of ovulation in pregnancy percentages in inseminated alpacas. The semen collection was made post copula. After the collection of the ejaculates, motility and volume were assessed. The ejaculates with volume (≥1 ml) and motility (≥ 60%) were mixed (pool), 80 % was used (3 samples/pool) and 20 % (2 samples/pool) during the experiment. Then and diluted in Tris-base with 20% egg yolk. The samples were cooled 1.5 hours at 5 °C. At that temperature it was combined with the basic dilutor plus glycerol, obtaining a final concentration of 5% glycerol, then they were packed in 0.5 mL (13 x 106 spem/straw) straws to be frozen by horizontal and vertical methods. The seminal (semen + vaginal fluid) quality analysis was performed fresh and after cooling and thawing. Insemination was performed in two groups with thawed semen from two straws of the vertical method, the first group 20 females induced to ovulate with GnRH analog, the second group 20 females induced to ovulate with seminal plasma. When comparing, the results obtained of motility, viability, HOST and acrosomal integrity of fresh sperm, after the freezing process, decreased (p<0.05) compared to fresh and refrigerated samples. On the other hand, when comparing the freezing methods, the sperm values frozen by the vertical method were higher than those obtained by the horizontal method, with (p<0.05) in motility and HOST without (p>0.05) in acrosomal integrity and viability. The vertical semen freezing method can replace the horizontal method to obtain pregnancy.


2016 ◽  
Vol 28 (2) ◽  
pp. 148
Author(s):  
M. L. Mphaphathi ◽  
M. M. Seshoka ◽  
T. R. Netshirovha ◽  
Z. C. Raphalalani ◽  
T. C. Chokoe ◽  
...  

Indigenous bulls semen are important for conservation programs. The objectives of this study were to evaluate the effects of repeated freezing and thawing on sperm motility characteristics. Semen was collected from 4 Nguni bulls by means of electro ejaculator and stored in a thermo flask (37°C). Sperm total motility, progressive and nonprogressive motility, and velocity were assessed using computer-aided sperm analysis before and after freezing. Semen was then diluted with egg yolk citrate extender (fraction A), then followed by 12% of glycerol + egg yolk citrate extender (fraction B, Seshoka et al. 2012). Diluted semen samples were equilibrated for 4 h at 5°C. After the equilibration period, samples were loaded into 0.25-mL straws and transferred into a controlled rate programmable freezer. After the target temperature of –130°C was reached, semen straws were stored in a LN tank (–196°C). After 3 months of storage, straws were thawed at 15°C (first and second freezing and thawing followed the same process) for 5 min and further evaluated post-thawed at 0 and 15 min during incubation at 15°C. Treatment means were separated using Fisher’s protected t-test least. No significant differences were recorded between the raw semen total sperm motility percentage (93.2%) and first frozen-thawed at 0 min (82.6%), with the total sperm motility rate recovery of 88.6%. In addition, there was a marked decline recorded in sperm total motility during the first frozen-thawed at 15 min (77.6%), second frozen-thawed at 0 min (31.3%), and second frozen-thawed at 15 min (30.1%; P < 0.05). The sperm curvilinear velocity and average path velocity was reduced following first frozen-thawed (P < 0.05) but remained constant and stable between the treatment groups (P > 0.05). In conclusion, the freezing-thawing process did not reduce the Nguni bull total sperm motility during the first freezing and thawing process, compared with raw semen. However, a drastic decline was recorded during the second freezing-thawing processes, compared with raw semen.


2018 ◽  
Vol 15 (2) ◽  
pp. 215-218
Author(s):  
S Deori

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)


2017 ◽  
Vol 18 (3) ◽  
pp. 345
Author(s):  
Magfira Magfira ◽  
Raden Iis Arifiantini ◽  
Ni Wayan Karniani Karja ◽  
Sri Darwati

The successful of artificial insemination (AI) depends on the semen quality and extender. To minimize effect of cold shock during storage, extender is added with egg yolk. The objectives of this study were to compare the effectiveness of pure Low Density Lipoprotein (LDL) and egg yolk from domestic chicken and quail on motility and longevity of Merawang chicken sperm. The semen was collected by massage method from three Merawang roosters. Immediately after collection, semen was evaluated macroscopically and microscopically. Only semen demonstrated >70% motility and <20% sperm abnormality were used in this study. Semen divided into four aliquots and diluted with Lactate Ringer (LR) LDL chicken (RL-LDL-C), LR-LDL quail (LR-LDL-Q), LR- chicken Egg Yolk (LR-CEY), Ringer Lactate quail Egg Yolk (RL-QEY). Diluted semen than stored at 5oC. Sperm motility was examined twice a day and the longevity of sperm was determined every day until the sperm reach 0% motility. The motility of spermatozoa in the LR-LDL diluent differed from the sperm motility in the RL-QEY diluent at the 60th and 72th hour (P <0.05) poststorage. However, there was no difference in motility sperm in LR-LDL-C, RL-LDL-Q and RL-CEY. Additionally, there is no difference (P> 0.05) in spermatozoa longivity in the four diluents, with a range of longivities between 4.43 to 5.93 days. ABSTRAK Keberhasilan inseminasi buatan (IB) salah satunya bergantung pada kualitas semen dan pengencer yang digunakan. Dalam meminimalisir pengaruh cold shock saat penyimpanan, pengencer ditambahkan dengan kuning telur. Tujuan dari penelitian ini adalah untuk membandingkan efektivitas Low Density Lipoprotein (LDL) dan kuning telur yang berasal dari ayam kampung dan puyuh terhadap motilitas dan longivitas spermatozoa ayam. Koleksi semen dilakukan menggunakan metode pemijatan pada tiga ekor ayam merawang. Setelah semen dikoleksi, selanjutnya semen dievaluasi secara makroskopis dan mikroskopis. Semen yang menunjukkan motilitas 70% dan abnormalitas kurang dari 20% dibagi empat dan diencerkan menggunakan Ringer Laktat-LDLA (RL-LDLA), Ringer Laktat-(RL-LDLP), Ringer Laktatkuning telur ayam (RL-KTA), dan RL-kuning telur puyuh (RL-KTP). Semen yang telah diencerkan kemudian disimpan pada suhu 5oC. Motilitas spermatozoa diamati dua kali sehari sampai motilitas mencapai 0%. Motilitas spermatozoa dalam pengencer RL-LDLA berbeda dengan motilitas spermatozoa dalam pengencer RL-KTP pada jam ke-60 dan ke-72 (P<0.05) pascapenyimpanan. Akan tetapi tidak terdapat perbedaan motilitas spermatozoa dalam RL-LDLA, RL-LDLP dan RL-KTA. Longivitas spermatozoa dalam empat pengencer tidak terdapat perbedaan (P>0.05) dengan rentang longivitas antara 4,43 sampai 5,93 hari.


Author(s):  
Abdullah Baharum ◽  
R. Iis Arifiantini ◽  
Tuty Laswardi Yusuf

The aims of this study were to investigate the freezing capability of Pasundan bull spermatozoa in Tris-egg yolk (TEY), Tris-soy (TS), and AndroMed® as diluents. Semen were collected twice a week from four Pasundan bulls aged 3-5 years old using an artificial vagina and evaluated macro- and microscopically. Semen had ≥70% sperm motility, ≥800x106/mL sperm concentration, and less than 20% sperm abnormalities were divided into three parts and each of them diluted with TEY, TS, or AndroMed®. After an equilibration step at 5°C for four hours, diluted semen were packaged in 0.25 mL straw, frozen in liquid nitrogen for ten minutes and kept in liquid nitrogen container until examination. Motility test on fresh, diluted, equilibrated, and after-thawed semen was done using Androvision®. The results showed that after thawing motility of sperm diluted in AndroMed® (58.64±0.72%) was higher than in TEY (49.45±1.22%) and TS (39.34±6.33%). Sperm motility of Pasundan bulls diluted in these three diluents reduced around 33.27±2.45% during freezing process.


2018 ◽  
Vol 12 (1) ◽  
pp. 86-91
Author(s):  
Nameer Al-biaty ◽  
Safaa Abed ◽  
Hazim J. Alobaidi ◽  
Safaa Almaliki ◽  
Abbas F Kareem ◽  
...  

The aims of the study was to investigate the possibility of using an plant lecithin; commercial soybean oil (SO) directly in the components of semen extenders of Awassi rams, storage of semen in chilled technique and the effects of dilution, cooling and storage periods on semen quality. Semen was collected weekly from four Awassi rams by electro- ejaculator. Pooled samples were divided to six equal aliquots and diluted by Citrate egg yolk extender at 37˚C. Treatments were designed on the base of control extender containing 25% egg yolk and four treatments containing different addition of SO (12.5, 25, 37.5, and 50%) and combination treatment of 12.5% egg yolk + 12.5% SO. Treatment tubes were cooled to 5˚C and stored for three days. Semen was evaluated as raw, diluted, cooled and after storage in refrigerator (5˚C) for 1, 2 and 3 consecutive days. Results showed that there were no significant differences among all treatments in progressive motility, dead sperm %, abnormal sperm % and acrosome defect %, while pH were found to be significantly declined (p≤ 0.05) in control group. Significant effects of dilution, cooling, and storage period have been demonstrated with steadily significant deterioration (p≤ 0.05) for all studied characteristics when motility and pH declined while sperm abnormality, dead, and acrosome defect percentages increased. The results clearly indicated successful of using different levels of SO (plant source) as lecithin source instead of egg yolk (animal source of lecithin) without any impacts on the biological characteristics of Awassi ram semen and the process of dilution, cooling and storage periods have deterioration effects on semen quality.


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