scholarly journals Identification of the Capacitating Agent for Bovine Sperm in Egg Yolk-TEST Semen Extender

1989 ◽  
Vol 72 (10) ◽  
pp. 2700-2706 ◽  
Author(s):  
A. Ijaz ◽  
A.G. Hunter ◽  
E.F. Graham
Keyword(s):  
Egg Yolk ◽  
2014 ◽  
Vol 26 (1) ◽  
pp. 203 ◽  
Author(s):  
M. E. Kjelland ◽  
T. Stroud ◽  
H. E. Ayliffe ◽  
G. Dittami

The primary objective of this research was to determine whether an automated electronic cell counter using microfluidic technology (Moxi Z) could be effective for determining sperm concentrations. Validation consisted of comparing frozen-thawed conventional bovine sperm counts using the Moxi Z to Coulter Z2 counter results. Specific goals were to determine (1) if the Moxi Z could count intact motile sperm, (2) if differences in media containing egg yolk lipids would affect counting, and (3) evaluate sample concentration requirements for Moxi Z accuracy. Cryopreserved straws (n = 20) of conventional bovine semen (Brangus), were obtained from Stroud Veterinary Embryo Services Inc. Straws were thawed (~30 s, 38°C) in a water bath, dried, and cut open. Semen was expelled into 15-mL conical tubes (BD Falcon) using a straw plunger. Thereafter, samples were centrifuged (400 × g, 10 min). The supernatant was removed and replaced with PBS (media 1) or Tris A semen extender (20% vol/vol egg yolk; media 2). Sperm samples exhibited ~70% motility for the experiments, based on subjective microscopy by placing 10-μL sample volumes on 25 × 75 × 1-mm microscope slides (#2951–001, Thermo Scientific, Waltham, MA, USA) with 24 × 50-mm #1.5 coverslips (#M6047–9, Baxter, McGraw Park, IL, USA) and viewing with a Axio Scope.A1 microscope (Zeiss, Thornwood, NY, USA) using 10× (M Plan Apo NIR, Mitutoyo Am, Aurora, IL, USA) and 40× (LD-Plan NeoFluor, Zeiss) lenses (i.e. 100–400×). The protocol for each sample was to establish an initial concentration of ~2.5 × 106 cells mL–1 and perform a serial dilution assay, using modified Hank's Balanced Salt Solution (Sigma, St. Louis, MO, USA). Counts (n = 3) for each dilution level (e.g. 1.0, 0.8, 0.6, 0.4, 0.2, 0.1, 0.05, 0.025) were made simultaneously on each instrument, Moxi Z (v3.6 OS, Orflo) and Coulter Z2 (Beckman Coulter, Fullerton, CA, USA). Data points were compared to linear fitted curves. The Moxi Z gated range was set at 2.88 to 6.54 μ to approximate the count range of the Z2 system (2.91–6.46 μ). The sperm samples provided similar results on both the Moxi Z and the Z2 instruments. Specifically, data for the 2 media using serial dilutions of the sperm samples were analysed, and the Moxi Z counts correlated well with the Z2 counts for media 1 (R2 > 0.9469) and media 2 (R2 > 0.9771). Overall the Moxi Z can provide rapid bovine sperm counts (<15 s) using small sample volumes. The degree of precision and accuracy depends strongly on proper gate setting and on the ability to reduce sample debris. Increased size resolution will be realised by future improvements to the Moxi Z signal processing (v 4.0), as well as by slowing the system flow rate; thereby improving cell peak discrimination. The portability of the Moxi Z may be an asset for field work and will be tested in future research. The data and dilution ranges herein demonstrate the utility of the Moxi Z for diluted conventional semen and lower dose concentrations, such as that of sexed semen.


2012 ◽  
pp. 7-10
Author(s):  
Csilla Budai ◽  
István Egerszegi ◽  
József Rátky ◽  
András Kovács

The aim of our study was to examine how different gelatin concentrations affect ram semens viability in liquid storage at 5 oC for five days. Our hypothesis was if we add gelatin to the semen extender, than the viability of ram semen will be better in the extenders containing gelatin, than the control. We used two different semen extenders:1.5% UHT milk and 1.5% UHT milk + 5% egg yolk. We added 0; 0.5; 1.0; 1.5; 2.0% Dr. Oetker gelatin to the semen extenders. We stored the semen for five days at 5 oC and in every 24 hour we made sampling.We stained the smears with Kovács-Foote staining and evaluated them with light-microscope. We categorized the cells in five groups like: live and intact cells, live cells with injured acrosome, dead cells, live head with dead tail and live tail with dead head. We used one-way analysis of variance (ANOVA) to assign how gelatin concentration affects the number of the categorized cells. On the fifth day, the viability was the best in the following semen extenders: 1.5% fat UHT milk + 1.0% gelatin and 1.5% fat UHT milk + 1.5% gelatin, but it was not significant (p>0.05).


2020 ◽  
Vol 25 (2) ◽  
pp. 68
Author(s):  
Nurul Afzan Hilda Zakiya ◽  
A H Yanti ◽  
T R Setyawati

The use of liquid semen for artificial insemination program of Etawah crossbreed goat (PE) is an alternative to replace frozen semen which is constrained by limited and expensive facilities. Production of liquid semen is faster than frozen semen, but the viability of liquid semen which preserved with a standard extender such as tris egg yolk is very short. The purpose of this study was to determine the viability of PE goat semen in egg yolk tris substituted with energy sources such as glucose, galactose, and mannose and to determine the most efficient energy source for semen preservation. This research was conducted from August to September 2018 at the Artificial Insemination Center in Lembang, West Java. This study was designed in a randomized block design (RBD) consist of three experimental groups divided into five groups. Fresh semen of PE goats were preserved using extender which energy source has been modified. Results showed that using glucose in PE goat semen extender produced the best motility among other groups (64.29 ± 9.2%). The highest viability was found in extender with fructose substitution (86.76 ± 2.3%). The longest viability of liquid semen was found in the extender with glucose substitution. It lasted for six days.


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.


2010 ◽  
Vol 22 (1) ◽  
pp. 337
Author(s):  
J. L. Anema ◽  
J. K. Graham ◽  
R. W. Lenz ◽  
G. E. Seidel

The objective of this study was to optimize bovine sperm storage for up to 20 h between semen collection and sex sorting followedby cryopreservation. Two successive ejaculates were obtained from mature dairy bulls (Holstein, n = 5; Jersey, n = 3) via artificial vagina. Treatments were then applied to the neat semen to which antibiotics were added as recommended by Certified Semen Services (Columbia, MO). Nothing further was added to the control samples until staining with Hoechst 33342 for sorting. For Treatment 1, semen was diluted 9:1 with a MOPS solution resulting in 24 mM MOPS and similarly, Treatment 2 resulted in 24 mM MOPS +2% egg yolk. A subsample of each treatment and control was sorted by flow cytometry shortly after collection, and sperm then were frozen following standard processing procedures. The other subsample was stored at 15-18°C and sorted 20 h after collection followed by cryopreservation. pH measurements were made before staining samples for sorting. Samples were evaluated post-thaw for subjective progressive and total sperm motility, by computer-assisted sperm analysis (CASA, Berkeley, CA, USA), and by flow cytometry for sperm viability using propidium iodide and SYBR-14. Treatment 1 performed better than the control (Table 1), while results for Treatment 2 were similar to the control. Second ejaculates were superior to first ejaculates. pH measurements showed that addition of MOPS kept the pH about 0.2 units higher than the control, but pH declined similarly over time in all groups. While responses for the 20 h sort were numerically lower than the 0 h sort (P > 0.1), the majority of responses were acceptable for most, but not all bulls. In conclusion, storing sperm in 24 mM MOPS was beneficial. Surprisingly, 2% egg yolk negated the beneficial effect of MOPS, possibly due to increasing osmolarity by ∼15mOsM/kg due to pH adjustment. Addition of MOPS provided better results than the control for both the 0 h and 20 h sorts. Table 1.Main effect means of semen characteristics


2006 ◽  
Vol 49 (1) ◽  
pp. 73-77 ◽  
Author(s):  
Alexandre Ninhaus-Silveira ◽  
Fausto Foresti ◽  
Yara Aiko Tabata ◽  
Marcos Guilherme Rigolino ◽  
Rosicleire Veríssimo-Silveira

Cryopreservation of semen from sex-reversed females of rainbow trout aims at rationalizing the production of stocks composed by 100% females. Semen from normal males (M) and two types of genotypic females (R and G), sex-reversed by the oral administration of 17alpha-methyltestosterone, were used. R was obtained by the fertilization of normal eggs with semen of sex-reversed females while G via gynogenetic reproduction. Semen was diluted in an extender solution (glucose 5,4 g, egg yolk 10 ml, dimetil sulfoxide 10 ml, water 80 ml) at 1:3 ratio (semen/extender), stored in straws of 0.5 ml and freezed in a dry container Cryopac CP-65, at -180ºC. Thawing was performed with water at 70ºC for 3 seconds. There were no significant fertilization rate differences (P>0.05) among thawed semen groups (M = 73.1±11.5%; R = 67.2±23.6%; G = 64±5.8%), confirming that the freezing methodology used was efficient to cryopreserve semen of all three trout groups.


2018 ◽  
Vol 51 (1) ◽  
pp. 21-28 ◽  
Author(s):  
Sakirat Opeyemi Adeyanju ◽  
James Olatinbo Daramola ◽  
Jimoh Alao Olanite ◽  
Olufiropo Samson Awokola

Abstract Soybean lecithin had been used as an alternative to egg yolk in domestic animal semen extender during cryopreservation due to its characteristic phospholipid content which played a major cryoprotective role. This composition of soybean lecithin informed the replacement of soybean with sunflower lecithin (SL) in the extender for the Kalahari Red (KR) buck semen cryopreservation in this study. Effect of different levels of SL on the quality of the KR buck semen during cryopreservation using slow freezing method was evaluated. Semen samples were collected from four KR bucks of between two and two and half of age using artificial vagina, evaluated for motility and then diluted in extenders containing different levels of SL (1.5%, 3.0% and 4.5%) as experimental group and 0% SL or 20% egg yolk as control. Semen parameters including motility, acrosome integrity (AcI), membrane integrity (MI), malondialdehyde (MDA) concentration, cholesterol level and seminal arginase activity were evaluated for. The results showed that motility, acrosome integrity (AI) and membrane integrity were comparable at 0%, (22.00 ± 4.58, 82.00 ± 3.51 and 96.00 ± 2.03); 1.5%, (23.00 ± 2.08, 87.00 ± 3.79 and 89.00 ± 2.08); 3.0%, (13.00 ± 2.52, 81.33 ± 0.41 and 76.67 ± 1.20) and 4.5% (11.00 ± 4.51, 85.33 ± 9.88 and 84.00 ± 8.50), respectively, after thawing. SL at 0% had the highest (P < 0.05) values for MDA, cholesterol and seminal arginase activity (1.10 ± 0.008 nmol/ml, 236.35 ± 4.08 mg/dl and 0.54 ± 3.3 E-3 units/mg protein, respectively). Our data suggest that 1.5% sunflower lecithin can be used in place of soy lecithin as a substitute for egg yolk during the cryopreservation of caprine semen.


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