<div style="text-align: justify;">This study was undertaken on three mature bulls each of Gir cattle and Surti buffalo breeds to evaluate the comparative efficacy of egg yolk based standard TFYG (Tris-citrate-fructose-yolk-glycerol) extender and egg yolk free soybean based commercial extenders Optixcell&reg; (IMV, France) and Andromed&reg; (Minitube, Germany) under split-sample technique. The ejaculates (9/bull) were extended @ 100&times;106 sperm ml-1 with three extenders and frozen using biofreezer following 4 hr of equilibration. The pooled means of progressively motile sperm observed (irrespective of extenders) at initial, pre-freeze and post-thaw stage in Gir bulls semen were 76.53&plusmn;0.53, 71.11&plusmn;0.53 and 39.86&plusmn;0.90% and in Surti buffalo 80.76&plusmn;0.39, 74.65&plusmn;0.45 and 40.35&plusmn;1.07%, respectively. The corresponding values for live sperm were 75.64&plusmn;0.76, 69.01&plusmn;0.97 and 47.99&plusmn;1.11 % for Gir and 80.90&plusmn;0.45, 75.76&plusmn;0.48 and 52.33&plusmn;0.86 % for Surti buffalo; and those of intact acrosome 94.29&plusmn;0.25, 90.29&plusmn;0.27 and 79.29&plusmn;0.33 % for Gir bulls, and 93.94&plusmn;0.21, 89.94&plusmn;0.23 and 78.95&plusmn;0.26 % for Surti buffalo semen, respectively. The HOS reactive sperm at initial, pre-freeze and post-thaw stage were 76.18&plusmn;0.74, 71.04&plusmn;0.76 and 27.90&plusmn;0.70 % for Gir, and 81.83&plusmn;0.35, 76.47&plusmn;0.39 and 27.83&plusmn;0.68 % for Surti bulls, respectively. The overall mean post-thaw incubation (37&deg;C) survival of spermatozoa observed at 60, 120 and 180 min were 28.40&plusmn;0.91, 17.78&plusmn;0.86 and 9.44&plusmn;0.72% for Gir bulls semen, and 28.01&plusmn;0.99, 18.40&plusmn;1.01 and 10.51&plusmn;0.93% for Surti buffalo semen, respectively. Optixcell was proved superior, and at par with TFYG, than the Andromed in maintaining greater motility, viability, morphology, acrosomal/plasma membrane integrity including post-thaw sperm longevity of cattle and buffalo spermatozoa with significant differences only in sperm motility and post-thaw longevity. The motile, live and HOST reactive sperm were significantly higher in buffalo semen than cattle at initial and pre-freeze stage, but not at post-thaw stage. The results showed that egg yolk free commercial Optixcell extender and egg yolk based TFYG extender were at par in terms of most of the sperm quality traits, hence any one of them can be preferred over Andromed for successful routine cryopreservation of cattle and buffalo semen.&nbsp;</div>

<div style="text-align: justify;">Efficacy of egg yolk based and egg yolk free soya bean milk based extenders for cryopreservation of Zebu cattle and buffalo semen</div>

Background: Semen extenders are crucially important when freezing animal semen to protect against sperm cell death and preserve the quality of the sperm cells. Semen diluents play a crucial role in the production process of freezing semen. Both pre-and post-freezing methods can be employed until the point of insemination. 
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Methods: This study aimed to compare the quality of sperm motility after freezing a newly produced semen Extender (MJ Ex2&trade;) with a conventional formula (Tris-egg yolk) using Computer Assisted Sperm Analysis (CASA). This was achieved by dividing the semen into two portions, diluting each portion with a different formula and then freezing both semen specimens using the same process. 
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Result: The results indicate that the fresh goat semen samples in this study had a cream color, an average volume of 1.10&plusmn;0.87 ml/time, a density of 4D (thick), a viscosity of 4oC (milky), a pH of 6.53&plusmn;0.14, a concentration of 17,203&plusmn;275.18 &times; 106/ml and a motility of ++++ (75.7&plusmn;0.13). The MJ Ex2&trade; extender diluent had a movement percentage of 76.72&plusmn;5.00, which was significantly higher than the Tris-egg yolk formula of 66.48&plusmn;2.89&nbsp; (P&lt;0.01). As for the percentage of forward movement, VAP, VSP, ALH and BCF values were not different. The motility characteristics of the Tris-egg Yolk formula had a LIN value of 50.19&plusmn;4.05, which was significantly high (P&lt;0.01). The percentage of motility and STR values of sperm diluted with MJ Ex2&trade; extender was 78.14&plusmn;5.04, which was significantly higher than the Tris-egg yolk formula 72.60&plusmn;7.93 (P&lt;0.05). The results of this study indicate that the MJ Ex2&trade; extender formula enhanced the quality of frozen goat semen. Considering the motility of the sperm; however, would suggest that other effects on those sperm cells would need to be further s to investigate.

Effects of Frozen Semen Extender for Goats (MJ Ex2&trade; and Tris-egg Yolk) on Sperm Motility Quality after Freezing

<div>Background: Taurine supplementation in tris egg yolk citrate (TEYC) extender can improve antioxidant defense and reduce motility degeneration rate in semen of Surti buck preserved at refrigeration temperature. Present study has evaluated antioxidant effect of different concentrations of taurine in TEYC extender on oxidative stress, antioxidant defense and motility degeneration rate in Surti buck semen preserved at refrigerated temperature. 
 
Methods: Four Surti bucks (age&gt;2years) were selected. Total 64 ejaculates (16 per buck) were collected during a period of 8 week. Post-collection they were pooled and stored at refrigeration temperature after dividing into 5 groups based on different taurine concentrations in TEYC extender viz. 0 mM (control T1), 25 mM (T2), 50 mM (T3), 75 mM (T4) and 100 mM (T5) taurine maintaining final concentration of 200 x 106 sperm/ml (pH 6.5-6.8). Evaluation of motility degeneration rate at 24, 36 and 48 hours and glutathione as well as lipid peroxidation (MDA) at 0 and 48 hours was done. 
 
Result: Supplementation of taurine @50 mM in TEYC extender caused post-chilling significant increase in reduced glutathione (GSH) levels, lowering of lipid peroxidation (in terms of MDA production) and reduction of motility degeneration rate (MDR)% in semen of Surti bucks. It improved antioxidant defense thereby maintaining good quality of semen.&nbsp;</div>

​Evaluation of Antioxidant Potential of Taurine in Tris Egg Yolk Citrate Extender in Surti Buck Semen Preserved at Refrigerated Temperature

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 &plusmn; 2kg, age=12 &plusmn; 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 &times; 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&gt;0.05) difference in sperm motility and viability was found among all groups. Sperm abnormality was significantly (P&lt;0.05) higher in G4 as compared to other groups (G1, G2, G3). The HOS response and acrosomal integrity was significantly (P&lt;0.05) higher in G1, G2 and G3 as compared to G4. However, no significant (P&gt;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&lt;0.05) higher in G1 and G2 as compared to G3 and G4. Sperm abnormality was significantly (P&lt;0.05) lower in G2 as compared to other groups. The acrosomal integrity was significantly (P&lt;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.&nbsp;

Effect of different levels of egg-yolk on freezability of Jakhrana buck semen 
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<div>Background: Egg yolk (EY) is well known to be toxic for buck spermatozoa, which creates restrictions of its use in cryopreservation. Therefore, this study is to compare the effect of different levels of gum Arabic (GA) in an extender on quality and fertility of cryopreserved buck sperm. 
 
Methods: Each ejaculate of six bucks was frozen in Tris with one of concentrations of GA which contained 3, 6, 9 and 12 gm/ 100 ml in place of the EY. Control was Tris extender containing 2.5% of EY.&nbsp; 
 
Result: A percentages of total motile sperm (54.92%; P&lt;0.05) and progressively motile sperm (26.22%; P&lt;0.05) of semen was frozen in Tris containing 9% of GA. Similar to control group, the pregnancy rate of does inseminated with extender containing 9% (50.0%) were significantly higher than those of does inseminated with extender containing 6% (8.33%), 3% (0.0%) and 12% (0.0%). Semen evaluation and fertility rate were similar when replacing the EY with GA in the Tris cryodiluent, after cryopreservation of buck semen. The present study shows that high motility rate of frozen semen and acceptable pregnancy rate can be obtained when using GA in place of EY for cryopreserving the buck sperm.</div>

Using Gum Arabic in Place of Egg Yolk as Cryoprotectant for Cryopreservation of the Buck Semen

Present investigation was carried out to study the bacterial load of different semen extender in frozen thawed semen samples of Sahiwal bulls maintained at Artificial Breeding Research Centre, ICAR-NDRI, Karnal, Haryana. Twenty four ejaculates from four bulls were collected during morning hours using sterilized artificial vagina and the ejaculates were evaluated. Semen samples showing more than 70 per cent progressive motility and having more than 800 million sperms per ml were used for further processing and freezing. The samples were split into six aliquots and extended in six different extenders [Conventional egg yolk (CEY), egg yolk of hen supplemented with omega enriched diet (group I), Egg yolk of hen supplemented with herbal feed (group II), 1% soya lecithin (SY1), commercially available Bioxcell (SY2) and Optixcell (LP)]. One ml of post thaw semen (four straws) was serially diluted (ten-fold) with normal saline, loaded in sterilized nutrient agar plate and incubated at 37&deg;C for 48 hours. Number of colonies were counted in duplicate and multiplied by dilution factor. The Standard plate counts (CFU/ml) of frozen thawed semen samples extended in different extenders were analyzed by one way analysis of variance. The results revealed that bacterial load of SY1, SY2 and LP was significantly (p&lt;0.05) different from egg yolk based extenders. The highest microbial load of frozen thawed semen samples was recorded in group II (646.30&plusmn;71.65) egg yolk extender and lowest in optixcell (91.67&plusmn;6.72) extender. Microbial load in all the extenders are well within the standard limit (5000CFU/ml) of frozen thawed semen sample. Lower bacterial load in present finding may be due to maintenance of better HACCP protocol for hygiene during preparation, semen collection and semen processing.

Microbial load of frozen thawed Sahiwal semen extended in egg yolk, soya lecithin and liposome based extender&nbsp;

<div style="text-align: justify;">The present study envisaged the effects of different levels of dietary copper supplementation on performance, egg quality parameters, serum and yolk cholesterol and triglyceride levels, and yolk fatty acid profile in laying hens. Total 96 Lohman laying hens (38 weeks old) were randomly divided into four groups (n=24 each). Experimental diets were prepared by adding copper (0, 200, 250 or 300 mg/kg) to basal diet and feeding was lasted 12 weeks. Dietary copper supplementation decreased egg weight and feed intake but did not affect other performance parameters compared to the control diet. Shape index, shell weight, shell thickness, yolk color, yolk index, albumen index, and haugh unit were not influenced by dietary copper supplementation, whereas shell strength increased due to the inclusion of copper in the basal diet. Copper supplementation did not affect the fatty acid profile of the egg yolk. The yolk cholesterol ratio decreased by 7.22% in the presence of 250 mg/kg copper supplementation compared to control diet. Therefore, high levels of dietary copper supplementation in the diet of laying hens can be beneficial for improving shell strength and reducing the cholesterol content of the yolk.&nbsp;</div>

Effects of dietary copper supplementation on performance, egg quality parameters, yolk cholesterol and fatty acid profiles in laying hens

Background: Cryopreservation of bovine semen is essential in artificial insemination programs. Traditional extenders using egg yolk and fructose have limitations in biosecurity and performance. Plant-based alternatives like soy lecithin and longan sugar may offer improvements. 
 
Methods: This study evaluated the effectiveness of extenders by replacing 20% egg yolk with 1% soy lecithin and substituting fructose with longan sugar. Semen from Angus, Charolais and tropical crossbred bulls was divided into four treatments: T1 (Egg yolk + fructose), T2 (Egg yolk + longan sugar), T3 (Lecithin + fructose) and T4 (Lecithin + longan sugar). Post-thaw semen quality was assessed using computer-assisted sperm analysis (CASA) in a 2&times;2 factorial randomized complete block design. Conception rates were recorded after artificial insemination. 
 
Result: Sperm quality in T1, T2 and T3 was significantly higher than T4 (P&lt;0.05). Kinematic parameters showed no significant differences among T1-T3 but were superior to T4 (P&lt;0.05). Lecithin groups showed more morphological abnormalities than egg yolk groups (P&lt;0.05). T4 was excluded from insemination due to substandard post-thaw quality. Pregnancy rates were highest in T3 (67.21%), followed by T1 (53.29%) and T2 (32.73%) (P&lt;0.05). The T3 extender (lecithin + fructose) maintained high semen quality and achieved the best conception rate. Lecithin combined with longan sugar is not recommended due to adverse interactions affecting sperm preservation.

Using Lecithin and Longan Sugar Instead of Egg Yolk and Fructose in Extender for Bull Semen

The aim of this study was to compare commercially available soybean based extenders, viz. Bioxcell&reg; and Optixcell&reg; (IMV, France) with standard Tris-citrate-fructose-egg yolk-glycerol (TFYG) extender for refrigeration preservation of buffalo semen. Forty eight good quality ejaculates (8/bull) having initial motility &gt;70% obtained at weekly interval in AV from 6 mature Surti buffalo bulls were split-diluted (at 34&deg;C keeping 100&times;106 sperm ml-1) in TFYG, Bioxcell and Optixcell extenders and stored in refrigerator at 5&deg;C for 72 hrs. The mean percentages of progressively motile spermatozoa observed on dilution (0-hr), and after 24, 48, 72 hrs of refrigeration preservation of semen at 5&deg;C were 78.54&plusmn;0.30, 68.30&plusmn;0.30, 59.38&plusmn;0.35 and 51.63&plusmn;0.47; live sperm 90.48&plusmn;0.19, 76.89&plusmn; 0.51, 69.51&plusmn;0.35 and 61.86&plusmn;0.45; morphologically normal sperm 93.85&plusmn;0.08, 90.70&plusmn;0.11, 89.08&plusmn;0.10 and 87.66&plusmn;0.10; intact acrosome 94.40&plusmn; 0.12, 88.22&plusmn;0.13, 84.60&plusmn;0.14 and 81.70&plusmn;0.17, and HOS reactive sperm 79.35 &plusmn;0.24, 65.99&plusmn;0.31, 57.69&plusmn;0.36 and 50.70&plusmn;0.43, respectively, all of which decreased gradually and highly significantly (P&lt;0.01) with increase in storage time. The results were significantly (P&lt;0.05) superior with Optixcell followed by TFYG extender than with the Bioxcell. The overall mean percentages of sperms with abnormal head, mid-piece and tail and those with different forms of acrosomal defects like swollen, ruffled, detached and denuded acrosome also followed the pattern of morphologically normal sperm and intact acrosomes. The values of all these traits increased significantly (P&lt;0.05) with increase in storage duration at 5&deg;C. The sperm protective ability of Optixcell extender was superior followed by TFYG and the least of Bioxcell. The Optixcell, a recently launched improved soybean milk based extender, excelled the Bioxcell extender and was statistically similar to TFYG in respect of all the sperm quality parameters to satisfactorily preserve the quality of buffalo semen at refrigeration temperature up to 3 days. 
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Evaluation of comparative efficacy of egg yolk and soya based extenders&nbsp;for refrigeration preservation (5&ordm;C) of buffalo semen

<div style="text-align:justify">Background: In order to find an alternative cryoprotectant for canine semen, different cryoprotectants, as dimethyl sulphoxide and ethylene glycol, dimethyle formamide either alone or in combination with glycerol have been evaluated on pooled semen of different breeds with different opinions. 
 
Methods: In the current study the effect of glycerol, DMF and their combination in Tris-citric acid-fructose egg yolk plasma extender on motility, viability, plasma membrane integrity (PMI), acrosome integrity (AI), inner mitochondrial membrane potential (IMMP), reactive oxygen species (ROS) and antioxidant enzymes in frozen-thaw semen of pug breed was compared. 
 
Result: Values for motility, viability, (PMI), AI were significantly (p&lt;0.05) high in TCFEYP-G compared to TCFEYP-DMF and TCFEYP-G+DMF at post thaw. However, HIMMP and MIMMP/MDA were non-significantly (P&gt;0.05) high and low in TCFEYP-G compared to TCFEYP-DMF and TCFEYP- G + DMF at post thaw, respectively. MDA concentration was significantly (p&lt;0.05) low in TCFEYP-G compared to TCFEYP-DMF and TCFEYP-G + TCFEYP-DMF extenders. However, H2O2, scavenging capacity of spermatozoa, cryopreserved in TCFEYP-G was significantly (p&lt;0.05) higher than in TCFEYP-DMF and TCFEYP-G + TCFEYP-DMF. However, there was no significant (p&gt;0.05) difference in superoxide free radical scavenging activity and nitrite concentration of spermatozoa among the three extenders. Activity of GPX, SOD and Catalase was also significantly (p&lt;0.05) higher in TCFEYP-G compared to TCFEYP-DMF and TCFEYP-G+DMF. However, values were reverse for GRE activity. This study concludes that glycerol is a healthier cryoprotectant to protect canine spermatozoa from cryo-injury during freezing-thawing process.</div>

Efficacy of egg yolk based and egg yolk free soya bean milk based extenders for cryopreservation of Zebu cattle and buffalo semen

Effects of Frozen Semen Extender for Goats (MJ Ex2™ and Tris-egg Yolk) on Sperm Motility Quality after Freezing

Evaluation of Antioxidant Potential of Taurine in Tris Egg Yolk Citrate Extender in Surti Buck Semen Preserved at Refrigerated Temperature

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

Using Gum Arabic in Place of Egg Yolk as Cryoprotectant for Cryopreservation of the Buck Semen

Microbial load of frozen thawed Sahiwal semen extended in egg yolk, soya lecithin and liposome based extender

Effects of dietary copper supplementation on performance, egg quality parameters, yolk cholesterol and fatty acid profiles in laying hens

Using Lecithin and Longan Sugar Instead of Egg Yolk and Fructose in Extender for Bull Semen

Evaluation of comparative efficacy of egg yolk and soya based extenders for refrigeration preservation (5ºC) of buffalo semen

Comparing Glycerol with Dimethylformamide and Combination for Cryopreservation of Pug Dog Semen in Egg Yolk Plasma Containing Extender