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.

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.

21 EVALUATION OF SEMEN EXTENDERS FOR SHORT-TERM STORAGE OF RAM SEMEN AT 4°C

2017 ◽  
Vol 29 (1) ◽  
pp. 118
Author(s):  
M. Acharya ◽  
J. M. Burke ◽  
C. Hansen ◽  
R. W. Rorie
Keyword(s):  
Ethylene Glycol ◽  
Repeated Measures ◽  
Mixed Model ◽  
Egg Yolk ◽  
Computer Assisted ◽  
Dilution Factor ◽  
Sperm Analysis ◽  
Progressive Motility ◽  
Ram Sperm ◽  
Over Time

Preliminary studies found that progressive motility of ram sperm declined ~75% when stored at 4°C for 24 h, and continued to decline over time when using extenders supplemented with 5% egg yolk. The current study evaluated the effects of different combinations of extenders, ethylene glycol (EG), egg yolk, and penicillamine, hypotaurine, and epinephrine on ram sperm progressive motility during storage. Semen collected from 3 Katahdin and 2 Suffolk rams by electroejaculation was distributed across treatment combinations consisting of either TRIS citrate or milk extender supplemented with 5 or 20% (v/v) egg yolk, ± 1% ethylene glycol (EG) and ± 20 µM penicillamine, 10 µM hypotaurine and 2 µM epinephrine (PHE). For each semen collection, TRIS citrate extender was prepared from a 4× solution so that the TRIS, citric acid and fructose concentration were constant at 300, 94.7, 27.8 mM, respectively, regardless of semen dilution factor. A 4× milk extender was also used so that the extender contained 10% (w/v) milk powder, regardless of semen dilution factor. Both extenders were supplemented with 50 µg mL−1 of gentamicin. Semen was diluted in extender to a final concentration of 300 million sperm/mL in 1.5-mL tubes, and cooled to 4°C over a 2- to 3-h period. Semen was evaluated initially and daily for 3 days, using computer-assisted sperm analysis. Repeated-measures data were analysed using the mixed model (JMP 12.0 software; SAS Institute Inc., Cary, NC, USA) for main effects of extender, supplements, and their interactions. Nonsignificant interactions were removed from the model before reanalysis. Data are presented as LSMeans ± standard errors. Initially, sperm progressive motility averaged 41 ± 6.2% across treatments. After an initial decline, overall progressive motility did not change (P > 0.05) significantly (mean of 22.3 ± 1.6 and 23.05 ± 1.3% at 48 and 72 h, respectively). Over time and across treatment combinations, mean progressive motility was maintained to a greater extent (P < 0.01) by milk than TRIS-based extender (28.2 ± 1.1 v. 18.9 ± 1.1%, respectively). Across extenders, progressive motility of sperm was similar (P = 0.50) for 5 and 20% egg yolk (22.2 ± 1.4 v. 24.4 ± 1.4). Addition of 1% EG increased (P < 0.01) progressive motility (25.8 ± 1.05 v. 21.3 ± 1.05). Addition of PHE also increased (P < 0.01) progressive motility from 20.9 ± 1.04 to 26.3 ± 1.04%. There was an interaction between EG and % egg yolk, primarily due to an effect on sperm stored in TRIS citrate extender. Addition of 1% EG to extender containing 5% egg yolk improved (P < 0.01) progressive motility from 18.5 ± 1.5 to 26.9 ± 1.5%). Addition of 1% EG to TRIS citrate extender also increased (P < 0.05) progressive motility, from 14.6 ± 1.5 to 23.2 ± 1.5%. Results indicate that milk extender supplemented with 1% EG, PHE, and either 5 or 20% egg yolk is capable of maintaining progressive motility of ram semen at ~60% of its initial value when stored at 4°C for up to 72 h. Additional studies are needed to evaluate pregnancy rate after insemination of ewes with stored semen.

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.

362 STORAGE OF BOVINE SPERM FOR 20 H BETWEEN SEMEN COLLECTION AND SEXING

2010 ◽  
Vol 22 (1) ◽  
pp. 337
Author(s):  
J. L. Anema ◽  
J. K. Graham ◽  
R. W. Lenz ◽  
G. E. Seidel
Keyword(s):  
Flow Cytometry ◽  
Egg Yolk ◽  
Computer Assisted ◽  
Ph Measurements ◽  
Sperm Analysis ◽  
Bovine Sperm ◽  
Semen Collection ◽  
Dairy Bulls ◽  
And Control ◽  
Better Than

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

90 EFFECT OF EGG YOLK ON THE KINEMATICS AND ACROSOME MEMBRANE INTEGRITY OF COOLED-REWARMED CANINE SPERMATOZOA

2010 ◽  
Vol 22 (1) ◽  
pp. 204
Author(s):  
J. Dorado ◽  
M. J. Galvez ◽  
M. R. Murabito ◽  
S. Demyda ◽  
L. J. De Luca ◽  
...  
Keyword(s):  
Semen Analysis ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Computer Assisted ◽  
Mean Values ◽  
Head Displacement ◽  
Healthy Dogs ◽  
Acrosome Integrity ◽  
Digital Manipulation ◽  
Lateral Head

Tris-egg yolk-based diluents provide adequate cryoprotection for the sperm of most species. This study was conducted to compare the ability of Tris-glucose extender containing 2 different concentrations of egg yolk to maintain sperm motility and acrosome integrity of canine spermatozoa during 72 h of preservation. For this purpose, a total of 20 ejaculates from 4 clinically healthy dogs (2 Spanish Greyhound, 1 German Pointer, and 1 Crossbreed) were collected by digital manipulation. The sperm-rich fraction of each ejaculate was divided into 2 aliquots. Then, they were diluted in Tris-based extender and centrifuged at 700g for 8 min. Sperm pellets were resuspended in either Tris buffer added to 20% (EY20) or 10% centrifuged egg yolk (EY10) and cooled to 5°C over 72 h. The effects of these extenders on motility and acrosome integrity were assessed objectively using a computer-aided semen analyzer (Sperm Class Analyzer, Microptic SL, Spain) and Spermac® staining, respectively. Each cooled-rewarmed semen sample was evaluated after 24, 48, and 72 h of preservation. Sperm motion parameters shown by computer-assisted semen analysis (CASA) are progressively motile (PMS) and motile spermatozoa (MS), curvilinear velocity (CLV), average path velocity (APV), progressive speed (SLV), and lateral head displacement (LHD). Data were statistically analysed by ANOVA. Dependent variables expressed as percentages were arsine-transformed before analysis. Differences between mean values were evaluated by the Duncan method. Data were presented as mean ± SEM. Differences were considered significant when P < 0.05. Analyses were performed using the statistical package SPSS 12.0. A total of 98 172 motile sperm trajectories were analyzed by CASA: 52 259 in EY20 and 45 913 in EY10. After 24, 48, and 72 h of preservation, MS and PMS were statistically higher (P < 0.01) in EY20. No significant differences were found for LHD using either extender over a 72-h period. No significant differences were observed for CLV using either extender during the first 2 days. At Day 3, CLV data were significantly higher (P < 0.01) in EY20. Similarly, from Day 2, APV was significantly higher (P < 0.001) in EY20. After 24 h of preservation, SLV was statistically higher (P < 0.001) in EY10, whereas the opposite tendency was found at Day 3. No significant differences were observed for SLV using either extender after 48 h of preservation. During the first 2 days, acrosome integrity was statistically higher (P < 0.001) in EY20. At hour 72, higher acrosome integrity (P < 0.001) was observed in EY10. In conclusion, we have observed that the EY20 extender provided higher motility after 72 h of chilled preservation; however, the acrosome membrane integrity was better preserved in EY10.

scholarly journals Effect of the addition of sodium caseinate on the viability of cryopreserved buffalo semen

2020 ◽  
pp. 2209-2218
Author(s):  
Fernando Evaristo da Silva ◽  
Jaqueline Candido Carvalho ◽  
Camila de Paula Freitas Dell'Aqua ◽  
Frederico Ozanam Papa ◽  
Marc Roger Jean Marie Henry ◽  
...  
Keyword(s):  
Cell Damage ◽  
Membrane Integrity ◽  
Sodium Caseinate ◽  
Egg Yolk ◽  
Freezing Process ◽  
Computer Assisted ◽  
Semen Cryopreservation ◽  
Sperm Analysis ◽  
Frozen Semen ◽  
Buffalo Semen

The use of cooled semen in artificial insemination operations results in higher pregnancy rates than the use of frozen semen. This result seems to be related to the more severe damage triggered by the freezing process than that observed during refrigeration. Due to its ability to bind to sperm-binding proteins and calcium ions, sodium caseinate has been studied as a substance capable of preventing early sperm capacitation, a significant cause of the decreased pregnancy rate resulting from the use of frozen semen. The first objective of this study was to evaluate whether a commercial egg yolk diluent developed for frozen bovine semen could be used for buffalo semen cryopreservation; the second objective was to investigate the effect of this diluent in combination with sodium caseinate during the procedures of buffalo sperm cryopreservation using flow cytometry and computer-assisted sperm analysis. In the first part of the study, comparing the results of spermatic kinetics and plasma and acrosomal membrane integrity, it was observed that the freezing process resulted in more cell damage than the cooling process. In the second part of the study, no effects of the addition of sodium caseinate to the egg yolk diluent were observed. From the results of the present study, it was possible to conclude that the egg yolk-based diluent was suitable for buffalo semen cryopreservation and that the addition of sodium caseinate did not decrease the harmful effects related to seminal cryopreservation.

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 The Quality of Frozen-thawed Canine Semen With Respect to Semen Extender Composition and Sequence of Ejaculate Collection in Dogs

2016 ◽  
Vol 64 (1) ◽  
pp. 169-175
Author(s):  
Jiří Šichtař ◽  
Ondřej Šimoník ◽  
Petra Folková ◽  
Adéla Dokoupilová ◽  
Radko Rajmon ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Computer Assisted ◽  
Time Interval ◽  
Sperm Analysis ◽  
Semen Collection ◽  
Movement Characteristics ◽  
Semen Extender

The aim of this study was to evaluate the effect of clarified egg yolk addition to semen extender, and the semen collection sequence on the quality of frozen-thawed semen in dogs. Semen was collected from 6 dogs in a time interval of 24 hours. As parameter of the quality of frozen-thawed (F-T) semen, the motility by computer assisted sperm analysis (CASA) and plasma membrane integrity by hypo-osmotic swelling test (HOS) were evaluated. All kinematic parameters of sperm motility were higher in F-T samples containing the whole in comparison to the clarified egg yolk. The sequence of semen collection affected sperm movement characteristics of native as well as F-T semen, but it was not possible to determine whether the fresh semen from the 1st or 2nd collection is of higher quality. All motility parameters of sperms frozen with extender containing the whole egg yolk were significantly higher in the case of the 2nd collection. The situation was not so clear in the case of clarified egg yolk addition, but the velocity values were higher in F-T samples from the 2nd collection. In contrast to proven differences in motility, the effect of the addition of clarified egg yolk and the sequence of semen collection were not projected at all on the quality of plasma membrane of canine sperms evaluated by HOS test.

Computer assisted sperm analysis of fresh and frozen-thawed buffalo semen and their interrelationship

2016 ◽  
Vol 50 (1) ◽  
Author(s):  
J. B. Patel ◽  
A. J. Dhami
Keyword(s):  
Sperm Motility ◽  
Computer Assisted ◽  
Motile Sperm ◽  
Mean Values ◽  
Sperm Analysis ◽  
Head Displacement ◽  
Straight Line ◽  
Buffalo Semen ◽  
Live Sperm ◽  
Fresh Semen

Sixty semen ejaculates from 10 mature bulls, 5 each of Jafarabadi and Mehsana breed, were studied for sperm motility and velocity parameters of fresh and frozen-thawed spermatozoa using computer assisted sperm analyzer (CASA). The mean values of motile and progressively motile spermatozoa observed in fresh semen of Jafarabadi and Mehsana bulls (79.77±1.62 and 61.80±1.85, and 78.90±1.22 and 61.37±1.58%) were highly significantly (P<0.01) reduced (51.20±1.57 and 33.20±1.45, and 52.10±1.70 and 34.30±1.54 %, respectively) in post-thawed semen. The average path velocity, straight line velocity and curvilinear velocity (μm/sec) of spermatozoa of Jafarabadi and Mehsana bulls noted in fresh semen were also reduced highly significantly (P<0.01) in frozen-thawed semen. Among the other velocity parameters, amplitude of lateral head displacement (μm), elongation (%) and medium motile sperm (%) increased, while beat-cross frequency (Hz), straightness (%), linearity (%), sperm area (μm<sup>2</sup>) and rapidly motile sperm (%) decreased significantly in post-thawed sperms when compared with the fresh sperm of both Jafarabadi and Mehsana bulls. The initial motility and live sperm per cent were significantly correlated with CASA traits of fresh and frozen-thawed semen, and all the sperm motility and velocity traits of fresh and frozen-thawed semen assessed by CASA were significantly interrelated among both the breeds. The interrelationships were stronger in Mehsana bulls as compared to Jafarabadi bulls.

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