58 EFFECT OF MELATONIN IMPLANT ON BLANCA de RASQUERA BUCKS DURING SPRING ON SPERM MORPHOMETRY BEFORE AND AFTER CRYOPRESERVATION

2015 ◽  
Vol 27 (1) ◽  
pp. 122
Author(s):  
A. Tabarez ◽  
W. García ◽  
M. J. Palomo
Keyword(s):  
Egg Yolk ◽  
Sperm Head ◽  
Head Width ◽  
Computer Assisted ◽  
Sperm Cryopreservation ◽  
Sperm Analysis ◽  
Know How ◽  
Before And After ◽  
Sperm Morphometry ◽  
Computer Assisted Sperm Analysis

In order to improve sperm cryopreservation throughout the year and accelerate the process of preservation of this Catalonian goat breed in extinction danger, we proposed to assess the effect of melatonin implant application in Blanca de Rasquera males during spring on sperm head morphometry of fresh and thawed sperm. Therefore 8 bucks of 30 months old approximately were divided into 2 groups. In one of the groups, 2 melatonin implants (Melovine®, CEVA) were inserted into bucks 60 days before starting the collection of semen, and the other group was kept untreated. Briefly, fresh ejaculates from each group of 4 bucks were collected in spring, immediately mixed in equal quantities, and centrifuged twice (600 g for 10 min). Then the pellet was resuspended in a Tris-based medium containing 15% (v/v) of powdered egg yolk supplemented with 5% glycerol. Afterward, sperm samples were refrigerated at 5°C for 4 h before being frozen in LN vapour. Buck sperm head morphometry was analysed by computer-assisted sperm analysis (ISAS®) on fresh and thawed sperm previously stained with Diff Quick®. Data were analysed by GLM multivariate procedure (IBM SPSS, 2011; mean ± s.e., n = 6), showing significant differences among treatments in all the morphometric parameters except head perimeter and rugosity (Table 1). Our results suggest that melatonin application in bucks increases the ellipticity and elongation of fresh and thawed sperm, meanwhile the cryopreservation process reduces both parameters. Likewise melatonin implants increase significantly the head length only on thawed sperms as cryopreservation process increases the head width, area in sperms from implanted males and regularity only in sperms from nonimplanted bucks. These head changes on fresh and thawed sperm morphometry should be deeply investigated in order to know how they could affect sperm cryosurvival and fertility. Table 1.Effect of melatonin implant on Blanca de Rasquera bucks during spring on morphometry of fresh and thawed sperm This research was supported by INIA (RZ2009–00008–00–00), Generalitat de Catalunya (2009SGR0621 and CUR-DIUE), and FSE and Fundación Carolina.

74 EFFECT OF POWDERED EGG YOLK IN THE EXTENDER AND DONOR AGE ON FRESH AND THAWED SPERM MORPHOMETRY IN SMALL RUMINANTS

2015 ◽  
Vol 27 (1) ◽  
pp. 130
Author(s):  
M. J. Palomo ◽  
W. Garcia ◽  
A. Tabarez
Keyword(s):  
Small Ruminant ◽  
Small Ruminants ◽  
Egg Yolk ◽  
Sperm Head ◽  
Computer Assisted ◽  
Donor Age ◽  
Mean Values ◽  
Sperm Analysis ◽  
Sperm Morphometry ◽  
Ram Sperm

Our aim was to reduce heterogeneity and microbiological contamination risk on small ruminant semen cryopreservation by replacement of fresh egg yolk by pasteurized powdered egg yolk, assessing simultaneously the effect of the donor age (1 year v. 2 years old) on sperm head morphometry of fresh and thawed sperm. Briefly, fresh ejaculates from 8 rams and from 8 bucks were collected in autumn during 2 consecutive years. Immediately after collection, ejaculates from each species were mixed in equal quantities, and pooled semen was centrifuged twice (600 g for 10 min). Then the pellet was split into 2 aliquots and resuspended in an extender containing 15% (v/v) of fresh or powdered egg yolk supplemented with 5% glycerol in a Tris-based medium. Afterward, sperm samples were refrigerated at 5°C for 4 h before being frozen in nitrogen liquid vapours. Buck and ram sperm-head morphometry was analysed by computer-assisted sperm analysis (ISAS®) on fresh and thawed sperm previously stained with Diff Quick®, and the data were analysed using ANOVA (mean ± s.e., n = 6). From ram sperm studies, no differences were found between fresh and post-thaw sperm, neither between egg yolk-based extenders or donor ages, showing the following mean values of head length (8.4 ± 0.0 μm), width (4.9 ± 0.0 μm), area (34.2 ± 0.1 μm2), perimeter (23.4 ± 0.1 μm), ellipticity (1.7 ± 0.0), elongation (0.2 ± 0.0), rugosity (0.8 ± 0.0), and regularity (0.9 ± 0.0). Likewise buck semen did not show significant differences on sperm-head dimensions after cryopreservation, only on head-shape parameters as ellipticity, elongation, and regularity between fresh and thawed sperm from 2-year-old donors, independently of egg yolk used as cryoprotectant. However, the age of the buck had a significant effect on all assessed morphometric parameters in fresh and thawed sperms, except regularity (Table 1). Our results suggest that, from a morphometric point of view, the powdered egg yolk can be used satisfactory on small ruminant sperm cryopreservation, but in goats, the head changes due to the donor age should be considered. Table 1.Effect of fresh v. powdered egg yolk (EY) in the extender and donor age on fresh and thawed buck-sperm morphometry Research supported by INIA (RZ2009–00008–00–00), Generalitat de Catalunya (2009SGR0621 and CUR-DIUE), and FSE and Fundación Carolina.

scholarly journals Effects of Three Semen Extenders, Breeding Season Month and Freezing–Thawing Cycle on Spermatozoa Preservation of Portuguese Merino Sheep

Animals ◽  
2021 ◽  
Vol 11 (9) ◽  
pp. 2619
Author(s):  
Margarida Fernandes ◽  
Pablo Rodríguez Hernández ◽  
João Simões ◽  
João Pedro Barbas
Keyword(s):  
Breeding Season ◽  
Interaction Effect ◽  
Egg Yolk ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Progressive Motility ◽  
Before And After ◽  
Semen Processing ◽  
Computer Assisted Sperm Analysis ◽  
Fresh Semen

This study aimed to evaluate and compare the effect of three semen extenders (S-EXT) on 22 spermatozoa (SPZ) parameters (subjective and computer-assisted sperm analysis evaluations), before and after semen cryopreservation throughout different months of the breeding season in the Portuguese Merino breed. According to the multivariable model, the SPZ viability (alive %), kinetics subjective individual motility, total motility, total progressive motility and its subpopulations, and beat cross frequency) were higher in the egg yolk-based S-EXT improved by Estação Zootécnica National (Portugal) than in Ovixcell® or Andromed® extenders. All the differences were only observed in thawed semen, except for total motility and total progressive motility, in which Ovixcell® also showed the poorest results on fresh semen. An interaction effect between S-EXT and semen processing was observed on 72.3% (17/22) of the evaluated parameters, evidencing a variable cryoprotective action between S-EXT. The SPZ viability was poorer in the onset of the breeding season (end of April/early May) than in the previous middle breeding season (November/early December), suggesting the influence of a short anoestrous season on ejaculate quality, even though the volume and SPZ concentration of the ejaculates remained stable throughout the experiment. Additionally, S-EXT x semen processing x month interaction effect on 59.1% (13/22) of the evaluated parameters evidenced the importance of SPZ time collection in a natural environment to cryopreserve ram’s semen. We concluded that, overall, the egg yolk-based S-EXT provided a greater value to the cryopreservation of Merino rams´ semen. Nevertheless, the causes of the interaction effect between S-EXT, semen processing and/or month on several SPZ parameters should be addressed, including SPZ molecular research in new studies, in order to improve egg yolk-based as well as in egg yolk-free-based S-EXT.

scholarly journals Evaluation of Bali Cattle Semen Quality During Cryopreservation with Coconut Water-Based Extenders

2021 ◽  
Vol 10 (4) ◽  
pp. 329-334
Keyword(s):  
Semen Quality ◽  
Egg Yolk ◽  
Coconut Water ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Progressive Motility ◽  
Different Types ◽  
Before And After ◽  
Computer Assisted Sperm Analysis ◽  
Bali Cattle

The aim of this research was to evaluate the Bali cattle semen quality during cryopreservation with different types of extenders in term of live, total motility, progressive motility, and abnormality of post-thawed bull sperms. The treatments were AndroMed® (T0), Tris-based egg yolk diluent (T1), Tris-based coconut water diluent (T2), Coconut water egg yolk diluent (T3). Bulls’ semen was collected from two adult Bali cattle maintained at the semen production facility at Bali Artificial Insemination Center, Tabanan Bali. The age of the bulls were 6 years old. Sperm live, total motility, progressive motility, and abnormality were analyzed with computer assisted sperm analysis (CASA) post-dilution, before and after thawed. The study was replicated five times, and data were analyzed using analysis of variance (ANOVA). The results showed that AndroMed ® and Tris-based egg yolk had significantly higher sperm live, total motility, progressive motility and abnormalities of spermatozoa for post-dilution, after equilibration and post thawed than Tris egg yolk coconut water and Coconut water egg yolk diluent. It was concluded that AndroMed® and Tris-based egg yolk can be considered as the best suitable extender for Bali cattle sperm cryopreservation. Coconut water had a deleterious effect when supplemented with 20% in tris and egg yolk.

scholarly journals Evaluación de la motilidad espermática a través del sistema C.A.S.A de semen caprino criopreservado bajo diferentes medios diluyentes

Respuestas ◽  
2013 ◽  
Vol 18 (2) ◽  
pp. 16-27
Author(s):  
Leonardo Hernández-Corredor ◽  
Alexander Nivia-Osuna ◽  
Daniel Hernández-Villamizar ◽  
Jorge Alexander Rubio-Parada ◽  
Armando Quintero-Moreno
Keyword(s):  
Lateral Displacement ◽  
Egg Yolk ◽  
Computer Assisted ◽  
Complete Medium ◽  
Sperm Analysis ◽  
Straight Line ◽  
Nitrogen Vapor ◽  
Computer Assisted Sperm Analysis ◽  
The University ◽  
Casa System

 El estudio evaluó la motilidad espermática y su efecto postdescongelación en semen caprino, en dos medios comerciales (Andromed® y TwoStep®) y diferentes protocolos de congelación (medio completo, con adicción del 10% de yema de huevo, semen centrifugado y sobrenadante seminal), se utilizaron machos de la raza alpina de la Universidad Francisco de Paula Santander Ocaña, el semen fue colectado con electroeyaculador, una vez los medios terminados y parte de los contenidos seminales enteros o centrifugados mezclados, se estabilizó por 2 horas, se envasó en pajillas de 0,5 cc y se congela en vapores de nitrógeno por 10 minutos, las pajillas se llevaron al laboratorio de Andrología de la Universidad del Zulia y por medio del sistema C.A.S.A.(Computer Assisted Sperm Análisis) se evaluaron los parámetros de motilidad como velocidad curvilínea (VCL), velocidad rectilínea (VSL), velocidad lineal (VAP), índice de linealidad (LIN), índice de rectitud (STR), índice de oscilación (ALH), Amplitud media del desplazamiento lateral de la cabeza del espermatozoide (BCF), los datos fueron analizados por medio del procedimiento GLM de SAS versión 9.0; los mejores índices de motilidad (VCL, ALH, BCF) fueron expresados enel tratamiento de contenido seminal centrifugado en medio Andromed®. (p≤0,001))La mejor progresividad espermática (VSL,LIN,STR)se presentó el tratamiento de Semen completo de caprino, criopreservado en medio comercial TwoStep®. ABSTRACT  The study evaluated the effect sperm motility and sperm post-thawing in goats, two commercial means (Andromed ® and Two Step ®) and different freezing protocols (complete medium with 10% addition of the egg yolk, semen centrifuged supernatant and seminal ), we used males of the Alpine race of the University Francisco de Paula Santander Ocaña, semen was collected with electroejaculator once finished media and part of the whole and centrifuged seminal contents mixed, stabilized by two hours, packed in 0.5 cc straws and frozen in nitrogen vapor for 10 min, the straws were taken to the laboratory of Andrology at the University of Zulia and through CASA system (Computer Assisted Sperm Analysis) were evaluated motility parameters such as curvilinear velocity (VCL), straight line velocity (VSL), linear velocity (VAP), linearity index (LIN), straightness index (STR) Oscillation Index (ALH ) average amplitude of the lateral displacement of the sperm head (BCF), the data were analyzed by the GLM procedure of SAS version 9.0, the highest rates of motility (VCL, ALH, BCF) were expressed in the treatment of seminal content centrifugation Andromed ® medium. (p ≤ 0.001)) The best progressive sperm (VSL, LIN, STR) will present the full Semen treatment goats, cryopreserved at Two Step ® commercial medium.Keywords: semen, buck, Andromed, Two step.

scholarly journals Effect of two freezing extenders on characteristic of fresh and frozen-thawed semen in endangered Old Kladruber stallions – A pilot study

2017 ◽  
Vol 62 (No. 6) ◽  
pp. 227-233 ◽  
Author(s):  
J. Šichtař ◽  
A. Nehasilová ◽  
O. Šimoník ◽  
F. Bubeníčková
Keyword(s):  
Egg Yolk ◽  
Quality Parameters ◽  
Computer Assisted ◽  
Kinematic Parameters ◽  
Fluorescence Staining ◽  
Sperm Analysis ◽  
Sperm Characteristic ◽  
Computer Assisted Sperm Analysis ◽  
Fresh Semen

The aim of the study was to evaluate the effect of two different extenders on sperm characteristic before equilibration and post-thaw in the endangered Old Kladruber stallions. Also, the response of individual stallions to the extenders used was tested. Semen was collected from six stallions every other day within one week. After centrifugation of the collected sperm-rich fraction, the supernatant was removed and sperm pellets were divided to two aliquots; these were diluted either with Gent (Minitube, Germany) or privately manufactured lactose-EDTA-egg yolk extender (Lact). Three cryopreserved insemination doses (IDs) from each extender (Gent and Lact) were prepared for each stallion from one collection (108 samples from six stallions in total). As a parameter of quality, the motility (computer assisted sperm analysis), viability (fluorescence staining), and morphology (eosin/nigrosine staining) were evaluated after dilution with freezing extenders (fresh) and after thawing (frozen-thawed). The different effects of chosen extenders on the quality of fresh semen were only manifested in higher kinematic parameters of sperm when the Lact extender was used. However, in frozen-thawed samples, the Gent extender yielded significantly better results in all of the evaluated parameters. The representation of sperm subpopulation was significantly influenced by extender in fresh as well as frozen-thawed samples; moreover, we found a significant effect of freezing on the distribution of these subpopulations. The response of individual stallions to chosen extenders was evident in the different quality of fresh as well as frozen-thawed IDs; Gent extender yielded better frozen-thawed IDs. Based on our results, among others describing quality parameters of ejaculate in endangered Old Kladruber stallions, we can recommend using Gent extender for the production of frozen-thawed IDs.  

scholarly journals Characteristics of the post-thawed Balinese bull semen extended in three different extenders and equilibration times

2019 ◽  
Vol 44 (2) ◽  
pp. 135
Author(s):  
A. S. Amal ◽  
R. I. Arifiantini ◽  
M. A. Setiadi ◽  
S. Said
Keyword(s):  
Semen Analysis ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Computer Assisted ◽  
Equilibration Time ◽  
Osmotic Swelling ◽  
Sperm Analysis ◽  
Bull Semen ◽  
Sperm Survival ◽  
Computer Assisted Sperm Analysis

The objectives of the present study were to compare and determine the best post-thawed characteristics of balinese bull sperm cryopreserved in three different extenders; animal based (Tris-clarified egg yolk (Tris-cEY)), and non-animal based extenders (Bioxcell® (lecithin based) and Optixcell® (liposome based)) in combination with three different equilibration times (30 minutes, 2 hours, 4hours). Thirty six ejaculates were collected from six Balinese bulls and frozen in three extenders (Tris-cEY, Bioxcell® and Optixcell®) after equilibration in three different times (30 minutes, 2hours and 4hours). Computer-assisted sperm analysis (CASA), hypo-osmotic swelling test (HOST) and eosin nigrosin staining were used in the post-thawed semen analysis. There was a significant interaction between equilibration time and extender type for sperm motility, viability and membrane integrity. Thirty minutes equilibration time had the lowest values (P<0.05) for all the evaluated parameters independent of extender type. Overall, semen extended in Tris-cEY, Bioxcell® and Optixcell® were similarly better when equilibrated at 4 hours (P>0.05). Moreover, post-thawed semen which were extended in Optixcell® for 2 hours equilibration showed a better motility compared with the other extenders (P<0.05). In conclusion, two hours equilibration of semen with Optixcell® is sufficient for semen freezing. Four hours equilibration has the best sperm survival, independent of the extender type.

306 THE EFFECT OF BoviPURE GRADIENT ON BISON SPERM CRYOPRESERVATION

2010 ◽  
Vol 22 (1) ◽  
pp. 309
Author(s):  
O. A. Bogle ◽  
C. Lessard ◽  
R. B. McCorkell ◽  
T. Grafton ◽  
G. P. Adams
Keyword(s):  
Disease Status ◽  
Egg Yolk ◽  
Computer Assisted ◽  
Gradient System ◽  
Separation And Purification ◽  
Freezing And Thawing ◽  
Sperm Analysis ◽  
Progressive Motility ◽  
Before And After ◽  
Wood Bison

Endemic tuberculosis and brucellosis threaten Canada’s wood bison population. For the purposes of developing a procedure to harvest pathogen-free sperm from bison, a sperm separation and purification product, BoviPure, was tested to determine its effect on bison sperm viability before and after cryopreservation. BoviPure (NidaCon International AB, Mölndal, Sweden) is a density centrifugation gradient system that contains trypsin and is designed to separate motile from non-motile sperm and remove infectious pathogens. Spermatozoa were expelled from the caudal epididymis of bison testicles collected at slaughter and diluted in 3 to 5 mL of TCM-199 medium (Gibco, Burlington, Ontario, Canada). After 1 h of incubation at 37°C, 1 to 1.5 mL of diluted sample (approximately 300 × 106 sperm) was placed in (1) a BoviPure gradient containing trypsin (n = 18 samples), (2) a BoviPure gradient without trypsin (n = 10 samples), or 3) left untreated (control; n = 20). Gradients were centrifuged at 300g for 20 min. The sperm pellet was resuspended in 5 mL of TCM-199 medium and recentrifuged at 500 g for 10 min. Total and progressive motility were estimated using computer-assisted sperm analysis (CASA). Gradient-treated and untreated sperm samples were diluted in an egg yolk extender (Triladyl; Minitube Canada, Ingersoll, Ontario, Canada), placed in 0.5-mL straws (50-200 × 106 spermatozoa per straw), frozen, and stored in liquid nitrogen for a minimum of 24 h. Semen straws were thawed by plunging in a 37°C water bath for 30 s, and motility measurements were taken immediately by CASA. Total and progressive motility were compared among groups by analysis of variance. Before cryopreservation, total motility and progressive motility were not affected by gradient processing. Total motility was 85±5.1%, 80±4.8%, and 77 ± 3.7% for untreated, BoviPure with trypsin, and BoviPure without trypsin, respectively (mean ± SEM), and progressive motility was 74 ± 2.7%, 77 ± 3.8%, and 68 ± 4.7%, respectively. After freezing and thawing, total motility decreased compared to unfrozen samples (P < 0.0001) but was not different among groups (37 ± 3.6%, 33 ± 3.7%, and 29 ± 4.8% for untreated, BoviPure with trypsin, and BoviPure without trypsin, respectively). Similarly, progressive motility decreased compared to unfrozen samples (P < 0.0001) but was not different among groups (26 ± 3.6%, 21 ± 3.7%, and 14.5 ± 4.9% for untreated, BoviPure with trypsin, and BoviPure without trypsin, respectively). We conclude that the BoviPure gradient with or without trypsin does not influence total motility or progressive motility of bison sperm either before or after cryopreservation and has potential as a method of harvesting pathogen-free sperm from wild animals of unknown disease status. This study was supported by Canadian Adaptation and Rural Development in Saskatchewan.

scholarly journals Qualitative ultrastructural changes and morphometry of deccani sheep spermatozoa preserved with egg yolk citrate extender

2017 ◽  
Vol 9 (3) ◽  
pp. 1515-1521 ◽  
Author(s):  
Madishetti Rajashri ◽  
Komati Ramchandra Reddy ◽  
Gangineni Aruna Kumari ◽  
Nagireddy Nalini Kumari ◽  
Surabhi Kesharwani ◽  
...  
Keyword(s):  
Qualitative Evaluation ◽  
Egg Yolk ◽  
Sperm Head ◽  
Ultrastructural Changes ◽  
Head Width ◽  
Electron Microscopic ◽  
Main Site ◽  
Initial Dilution ◽  
Liquid Storage ◽  
Sperm Morphometry

The present investigation was aimed to study the sequential changes in the sperm cell deterioration dur-ing liquid storage of Deccani sheep breed semen from dilution to 48 h of storage along with its seminal characteris-tics and sperm morphometric measurement. Thus the two Deccani adult rams (aged 2 years), were selected (six ejaculates/each ram) and the collected semen was diluted with Egg yolk citrate extender (EYC) (final concentration - 400 million spermatozoa/0.2 ml semen).Seminal characteristics were assessed along with sperm morphologi-cal changes by Electron microscopy immediately after dilution, at 24 and 48 h of storage, respectively. Sperm morphometry was analysed by Image analysis.The percentage of Individual motility, Live spermatozoa, Acrosomal integrity and Hos-test reactive sperm decreased significantly (P<0.05) from 80.41 to 49.16%, 82.75 to 51.25%, 94.16 to 83% and 76 to 48.58%, respectively during liquid storage of semen from initial dilution to 48 h of storage. The sperm head length (μm), Head width, sperm head area (μm2), sperm head perimeter (μm), mid piece length (μm), proximal mid piece width (μm), distal midpiece width (μm), volume of mid piece (μm3) and acrosomal cap length (μm) were 7.80, 4.33, 26.84, 20.63, 14.03, 0.74, 0.51, 4.54 and 5.24, respectively. Electron microscopic qualitative evaluation revealed that the main site of injury is the apical ridge of ram spermatozoa when stored at 5ºC. The electron density of the mitochondria reduced indicating concomittant depletion of ATP and loss of motility resulting in reduction of fertility.

scholarly journals Effect of Cholesterol and Equex-STM Addition to an Egg Yolk Extender on Pure Spanish Stallion Cryopreserved Sperm

2013 ◽  
Vol 2013 ◽  
pp. 1-6 ◽  
Author(s):  
Lidia Gil ◽  
Iván Galindo-Cardiel ◽  
C. Malo ◽  
N. González ◽  
C. Álvarez
Keyword(s):  
Sperm Quality ◽  
Egg Yolk ◽  
Computer Assisted ◽  
Pig Liver ◽  
Sperm Analysis ◽  
Cryopreserved Sperm ◽  
Fluorescence Test ◽  
Acrosome Integrity ◽  
Computer Assisted Sperm Analysis ◽  
Economic Improvement

Cholesterol and Equex-STM are frequently added to different commercial and experimental extenders improving postthawing sperm quality. Doses of 125–150 mM of cholesterol from pig liver and 0.5–0.7% of Equex-STM were evaluated in a standard eggyolk extender (Martin et al., 1979). Six ejaculates per stallion from six pure Spanish stallions (6–8 years old) were collected in Martin's extender (B) and different mixtures of 125 mM-0.5% (I), 125 mM-0.7% (II), 150 mM-0.5% (III), and 150 mM-0.7% (IV) were added to original Martin's extender. Samples were frozen in 0.5 mL straws (100×106 spermatozoa) and thawed (21 s., 37∘C water bath). After thawing the following parameters were evaluated: viability (V), motility (computer assisted sperm analysis, CASA; % nonprogressive NP; % progressive MP), hipoosmotic swelling test (HOST), acrosome integrity (A), fluorescence test (FL), and resistance test (RT). Sperm quality was significantly affected by stallion (in the parameters V, VI, NP, MP, HOST, A, FL, and RT), extraction (VI, NP, MP, HOST, A, and FL), and the different combinations of Equex-STM-cholesterol (FL). We concluded that 0.5% of Equex-STM mixed with 125 mM of cholesterol has obtained better sperm quality results than those of original Martin's extender, showing a simple and economic improvement of this home-made practical seminal extender.

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