Egg Yolk-sodium Citrate Research Articles

The Effect of Additional Tomato Extract in Diluting Sodium Citrate and Egg Yellow in Kampung Chicken on The Quality of Semen Cow Limousine

Undiluted sperm will reduce fertility, therefore dilution is performed to maintain the quality of spermatozoa during storage. Tomatoes contain carbohydrates, vitamin A, vitamin C, protein, and lycopene as antioxidants. Carbohydrates and antioxidants act as a source of energy to potentially prevent free radicals that can damage cells. This study aims to determine the effect of adding tomato juice in sodium citrate diluent and free-range chicken egg yolks to the semen quality of limousin cattle. This study used a completely randomized design (CRD) with 4 treatments and 5 replications. The treatment consisted of a solution of sodium citrate and egg yolk (P0), sodium citrate and egg yolk + 10% tomato extract (P1), sodium citrate and egg yolk + 20% tomato extract (P2), and egg yolk sodium citrate + 30% tomato extract. % (P3). Parameters observed were motility, viability and abnormalities of semen spermatozoa of Limousin cattle. Observations were made at 0, 24, 48, and 72 hours after cooling. The results showed that the addition of 10% tomato extract (P1) had a significant effect on the highest percentage of motility, namely 42.00 ± 2.73c for the 48th hour, the highest viability 62.10 ± 4.51c for the 72nd hour, and lowest abnormality 16.60 ± 1.81a until the 72nd hour. The addition of 10% tomato extract (P1) in sodium citrate diluent and egg yolk resulted in the best motility, viability, and abnormality percentage of Limousin cattle spermatozoa.

Cryopreservation of Indigenous bull semen (Zobawng) of Mizoram: Effect of fructose on extender

A total of 36 ejaculates were collected from three indigenous bulls (Zobawng) of Mizoram. The semen samples were evaluated to find out the effects of fructose i.e. Tris egg yolk citric acid fructose glycerol (TEYCAFG) and Egg yolk sodium citrate glycerol (EYSCG) extenders on freezability of indigenous bulls’ (Zobawng) semen of Mizoram in terms of sperm motility, per cent live sperm, HOSST reacted spermatozoa and acrosomal integrity. The mean sperm motility, per cent live sperm, HOSST and acrosomal integrity in TEYCAFG extender at pre and post freezing were (69.53±0.31 and 45.36±0.59%), (76.36±0.35 and 53.03±0.45%), (67.91±0.45 and 44.92±0.56%) and (75.17±0.58 and 63.25±0.56%) respectively, and in EYSCG extender were (45.67±0.48 and 33.08±0.36%), (54.56±0.41 and 38.08±0.28%), (47.86±0.44 and 34.61±0.40%) and (57.22±0.31 and 45.89±0.48%) respectively. The mean motility, live sperm, HOSST and acrosomal integrity in both TEYCAFG and EYSCG extenders were significantly (P

Unilateral intrauterine horn insemination of frozen semen in cats.

Frozen feline semen was prepared using two types of extenders, egg yolk Tris-fructose citric acid (EYT-FC) and egg yolk sodium citrate solution (EYC), and the semen qualities after thawing and the conception rates obtained by unilateral intrauterine horn insemination (UIUI) were investigated. Cats used in the experiment were six males and 11 females aged 2-12 years (the number of experimental cases was 17). For preparation of frozen semen, semen collected by the artificial vagina method was adjusted to I x 10(8) sperm/m/ and 7% glycerol, put in 250 microl straws, and then frozen using a cell freezer. The mean sperm motility after thawing was 30.0+/-9.7 (SE) % in the semen prepared with EYT-FC and 30.0+/-3.3% in the semen prepared with EYC. Four of seven animals were fertilized by UIUI using two straws in both extenders, and the conception rate was 57.1%. The mean ratios of number of kits to the number of ovulations in the inseminated side were 61.1+/-24.5% and 30.5+/-3.4% for EYT-FC and EYC, respectively, showing that the ratio tended to be higher in the semen prepared with EYT-FC. The above findings, comparing the two extenders for preparation of frozen feline semen, showed that EYT-FC is slightly superior to EYC. To increase conception and fertility rates, it may be important to increase the sperm count for insemination and to inseminate both uterine horns.

A technique for the evaluation of sperm penetrating ability and quality of bovine semen processed in an extender made with brackett-oliphant medium and egg yolk

Egg yolk-sodium citrate (EYC) semen extender was compared with an extender made of Brackett-Oliphant medium and egg yolk (BOEY). Ejaculates were divided into equal portions, processed and frozen. Semen was thawed and evaluated for quality. Additional semen was thawed, stained with Hoechst 33342 and the spermatozoa capacitated, after which they were coincubated with zona-free hamster oocytes to determine their penetrating ability. Sperm penetration of non-compressed, unfixed oocytes was evaluated using an optical sectioning technique on a standard research microscope. Sperm penetration was considered successful if a fluorescing sperm head was observed within the living oocyte in a hanging drop of fertilization medium. There were small differences in percentage of secondary abnormalities and percentage of progressive motility immediately after thawing between spermatozoa extended in EYC or BOEY diluent. There were no differences due to by extender composition in percentage of spermatozoa with intact acrosomes or percent of progressively motile after a 3 h incubation at 37 °C, nor the percentage of spermatozoa with head abnormalities. While there were significant correlations between all seminal quality characteristics, no quality measurements were correlated to percentage of oocyte penetration. The new penetration evaluation method allowed for examination of the fertilized oocytes using fluorescent microscopy initially and again after re-incubation for further development.

Studies on lactic dehydrogenase and sorbitol dehydrogenase release in relation to deep freezing of buffalo semen in certain extenders

Forty seminal ejaculates from five mature buffalo bulls (n=8 from each bull), exhibiting more than 70% initial sperm motility, were frozen in the following three extenders: egg-yolk sodium citrate glycerol (EYCG); tris-egg yolk glycerol (TYG); and citric acid whey glycerol (CAWG). The extenders were evaluated for the release of intracellular enzymes, lactic dehydrogenase (LDH) and sorbitol dehydrogenase (SODH) from the spermatozoa during freezing (in fresh semen, after dilution and after equilibration) and post freezing (24 h and 7 d after freezing. It was found that the release of LDH and SODH enzymes was significantly lower in TYG than in EYCG and CAWG extenders. The most critical stage, at which the enzyme release was maximal, was between equilibration and 24 h post freezing in all three extenders.

A new antibiotic combination for frozen bovine semen, 3. Evaluation of fertility

Field fertility (nonreturn rate) studies were performed independently by three artificial insemination organizations to evaluate bovine semen processed for freezing using the antibiotics gentamicin, tylosin and Linco-Spectin at concentrations of 500 ug, 100 ug, and 300/600 ug, respectively, per milliliter of neat semen and per milliliter of nonglycerol portion of the extender. The antibiotic combination including penicillin, dihydrostreptomycin, polymyxin B sulfate, with/without Linco-Spectin (500 units/ml, 2000 ug/ml, 1000 units/ml and 300/600 ug/ml, respectively) was used as the control treatment. Results indicated no significant effect on seminal quality as measured by field fertility under the conditions of these experiments using heated whole milk or egg yolk-sodium citrate seminal extenders. Use of the new antibiotic combination has been adopted by Certified Semen Services.

Influence of Glycerol Concentration and Freezing Rate on Post-thaw Motility of Bovine Spermatozoa in Continental Straws

Semen from each of six bulls was used to study the effects of percent glycerol and rate of freezing on spermatozoa extended in egg yolk-sodium citrate extender and packaged in .25-ml Continental1 straws. Treatments were arranged factorially and included 5, 6, 7, 8, 9, 10, or 11% (volume/volume) glycerol; freezing rates (time from 5 to —130C) of 1, 2, 3.5, 7, 12, or 20min; and post-thaw incubation times of 0 and 2h at 38C. Post-thaw motility was similar for spermatozoa frozen in 2 to 20min, and all means were greater than for spermatozoa frozen in 1min. Post-thaw motility of spermatozoa frozen in 7% glycerol did not differ from that with 5 to 9% but was superior to that with 10 or 11% glycerol. Although glycerol and freezing rate interacted, 7% glycerol provided survival equal to or greater than that with the other percents of glycerol, regardless of freezing rate.

The freezability of spermatozoa after minimal pre-freezing exposure to glycerol or lactose

Bull spermatozoa were frozen in egg yolk-sodium citrate and egg yolk-Tris media after different lengths of exposure to a range of glycerol levels at 5 °C by placing 0.1 ml of extended semen in small holes on a dry ice block. Post-thaw motility was significantly greater after glycerol exposure of 10 sec before freezing (shortest time studied) as compared with 2 min of exposure or longer ( P < 0.05). After 6 min of glycerol exposure no further significant decline occurred. Thus, momentary exposure to glycerol provided the best conditions for sperm survival during freezing. It is proposed that immediately after glycerol addition dehydration of the spermatozoa occurs and by freezing the cells immediately at a rapid rate, intracellular ice formation and the prolonged influence of high solute concentration may be avoided. Similar studies were conducted by freezing sperm in the presence of a presumably nonpenetrating cryopreservative, lactose, without glycerol. Post-thaw motility was not influenced by the length of prefreezing exposure to lactose at 5 °C. The fact that only minimal exposure to both cryopreservatives is required for optimum survival supports the concept that penetration is unnecessary. The studies with glycerol suggest that penetration may be undesirable for bull spermatozoa.

A Comparison of Egg Yolk-Sodium Citrate, Egg Yolk-Sodium Citrate-Glycine, and Egg Yolk-Glycine as Semen Diluters

Summary The average length of time that a +3 motility was maintained in egg yolk-citrate-glycine, egg yolk-glycine, and egg yolk-citrate, was 14.7, 13.9, and 6.8 days, respectively, for bovine sperm; 6.8, 8.8, and 3.5 days, respectively, for buffalo sperm; and 17.2, 8.0, and 4.4 days for ram sperm. The egg yolk-sodium citrate-glycine mixture gave better visibility of the sperm in microscopic preparations.

The Effect of Glycerol Equilibration Time on the Freezing of Bovine Spermatozoa in Egg Yolk–Sodium Citrate and Skimmilk Semen Extenders1,2

The Effect of Glycerol Equilibration Time on the Freezing of Bovine Spermatozoa in Egg Yolk–Sodium Citrate and Skimmilk Semen Extenders1,2

Dilution of Bull Semen with Frozen Egg Yolk-Sodium Citrate

Summary By the use of a split-sample technique, the fertility of bovine semen was studied using four diluters: (a) freshly prepared control diluter, C; (b) freshly prepared diluter to which penicillin-streptomycin had beeen added, PS; (c) diluter prepared and frozen one month prior to use, FC; and (d) diluter to which penicillin-streptomycin had been added and then frozen one month prior to use, FPS . Results, based on 60- to 90-day non-returns to 1,558 first services were as follows: C, 60.5 per cent; PS, 58.3 per cent; FC, 61.8 per cent; and FPS, 63.3 per cent. An analysis of variance showed that there were no significant differences in conception rates.

The Effect of Varying Proportions of Egg Yolk and Sodium Citrate Buffer in Bull Semen Diluters upon Sperm Motility

Summary An investigation has been made of the egg yolk-sodium citrate buffer diluter for bull semen with the view of establishing optimum concentrations. It was found that 3 per cent sodium citrate dihydrate was superior to other concentrations tried, and this solution was nearly isotonic with fresh bull semen. Reducing egg yolk concentration to 20 per cent gave as good motility and livability as the commonly used 50 per cent. When egg yolk was reduced to 10 per cent the cold shock protective factor was still fully effective but motility was impaired slightly.