Semen quality and fertility of liquid stored and frozen-thawed semen in crossbred pigs of North-Eastern India

Study was conducted to compare the semen quality and fertility of liquid stored semen for three days and frozen-thawed semen in the north-eastern region of India. For liquid semen, the semen ejaculates were extended in Beltsville Thawing Solution (BTS) extender and preserved at 17°C for three days. For cryopreservation, semen was diluted Lactose-egg yolk-glycerol extender and frozen in straw using programmable freezer with freezing rate of 40°C/min from -6 to -140°C. The preserved evaluated for sperm motility, viability, plasma membrane integrity and fertility. The results revealed that the liquid stored semen has maintained the sperm motility and viability up to day 3 without significant reduction. Similarly the plasma membrane integrity did not differ significantly up to day 2, but it was significantly (P less than 0.05) reduced on days 3 in liquid stored semen. After freezing and thawing, the mean sperm motility, viability and plasma membrane integrity were 58.25 ± 2.96%, 64.75 ± 2.47% and 47.06 ± 2.02%, respectively. These parameters were significantly (PP less than 0.01) lower as compared to the liquid stored semen from day 0 to day 3. After insemination with liquid semen, the farrowing rate was 77.7%, 80.76%, 73.07% and 69.8%, respectively from day 0, day1, day 2 and day 3. The pregnancy rate, farrowing rate and litter size did not differ significantly among different days of liquid storage. These parameters were significantly (PP less than 0.01) lower in frozen semen as compare to that of liquid stored semen. The study concluded that the liquid semen stored up to three days is more efficient than frozen-thawed semen in terms of preserving sperm quality and fertility.

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Correlations Amongst Functional and Morphological Attributes and Oxidative Markers of Fresh and Cryopreserved Semen of Gir and Murrah Bulls

THE INDIAN JOURNAL OF VETERINARY SCIENCES AND BIOTECHNOLOGY ◽

10.21887/ijvsbt.15.3.7 ◽

2020 ◽

Vol 15 (03) ◽

pp. 24-29

Author(s):

A. J. Dhami ◽

Tapasvi M Patel ◽

DV Chaudhari

Keyword(s):

Plasma Membrane ◽

Sperm Motility ◽

Semen Quality ◽

Sperm Quality ◽

Membrane Integrity ◽

Winter Season ◽

Correlation Coefficients ◽

Plasma Membrane Integrity ◽

Murrah Buffalo ◽

Oxidative Markers

This study was undertaken during the winter season on healthy mature Gir cattle and Murrah buffalo bulls (n=3 each). The semen samples (6 ejaculates/bull, total 36 ejaculates) collected in the morning using artificial vagina were evaluated for routine seminal attributes, including acrosomal and plasma membrane integrity. The samples were then diluted @ 100 million sperm/ml with tris fructose yolk glycerol extender without and with sericin @ 0.1, 0.25, 0.5 and 1.0% (w/v), filled in French mini-straws, and frozen in LN2 using biofreezer as per standard freezing protocol. Straws were thawed in water bath at 37°C for 30 sec and evaluated for post-thaw quality, viz., motility, viability, morphology, acrosome integrity and plasma membrane integrity (HOST). Lipid peroxidation (malondialdehyde - MDA production) and activities of enzymes superoxide dismutase (SOD) and glutathione peroxidase (GPx) were assessed as oxidative markers in seminal plasma of freshly diluted and frozen-thawed semen samples. Sericn at 0.5% level significantly (p less than 0.01) improved the post-thaw sperm quality with reduced oxidative stress in both the species. The breed-wise correlation coefficients (r) among sperm quality attributes and oxidative markers were studied in fresh and frozen-thawed semen of each species, and also for fresh with frozen-thawed semen. The findings revealed significant interrelationships amongst most of the attributes of fresh as well as post-thawed semen and also of fresh semen attributes with those of cryopreserved semen including oxidative markers in both the species. Sperm motility estimation in fresh, pre-freeze and post-thawed semen was a legitimately good indicator of quality of spermatozoa at various steps of semen processing/freezing, and its fertilizing potential. Thus, the sperm motility, HOS test and either MDA or SOD/GPx activity alone may be used as valuable and practical tools for routine assessment of bovine semen quality considering significant correlations found between them.

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Cryoprotective Effect of Glycerol Concentrations on Indian Red Jungle Fowl (Gallus Gallus Murghi) Spermatozoa

Avian Biology Research ◽

10.3184/175815618x15180876264262 ◽

2018 ◽

Vol 11 (2) ◽

pp. 80-88 ◽

Cited By ~ 7

Author(s):

Bushra Allah Rakha ◽

Muhammad Sajjad Ansari ◽

Shamim Akhter ◽

Elisabeth Blesbois

Keyword(s):

Plasma Membrane ◽

Semen Quality ◽

Sperm Quality ◽

Membrane Integrity ◽

Gallus Gallus ◽

Recovery Rates ◽

Plasma Membrane Integrity ◽

Red Jungle Fowl ◽

Frozen Semen ◽

Acrosome Integrity

Semen cryopreservation protocols for wild avian species need to be optimised in order to achieve optimum post-thaw sperm quality and fertility. The present study was designed to evaluate the cryoprotective effect of different glycerol concentrations (11%, 15% and 20%) on post-thaw quality, recovery rates, absolute livability index and fertility of Indian Red Jungle Fowl (Gallus gallus murghi) semen. Semen was collected from eight mature cocks and cryopreserved for storage at −196 °C. Frozen semen was thawed at 37 °C for 30 s and assessed for motility, plasma membrane integrity, viability and acrosome integrity at 0, 2 and 4 h incubation at 37 °C. Percentages of motility, plasma membrane integrity, viability and acrosome integrity were recorded higher (P<0.05) post-thaw at 0, 2 and 4 h at 37 °C with 20% glycerol compared to 15% and 11% glycerol. Likewise, recovery rates (%) of aforementioned parameters after cryopreservation and absolute livability index were observed highest (P<0.05) with 20% glycerol. By comparing values of R2 after multivariate regression analysis, least negative effects of hours of incubation were observed on semen quality in extenders with 20% glycerol followed by 15% and 11% glycerol. The fertility outcomes (number of fertile eggs, fertility [%], number of hatched chicks, percent hatch and hatchability of fertilised eggs) were recorded higher (P<0.05) with 20% glycerol followed by 15% and 11% glycerol. It is concluded that the concentration of 20% glycerol gives the best cryoprotection for quality and fertility of Indian Red Jungle Fowl semen.

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The effects of chronic administration of ghrelin on rat sperm quality and membrane integrity

Animal Biology ◽

10.1163/157075609x437682 ◽

2009 ◽

Vol 59 (2) ◽

pp. 159-168 ◽

Cited By ~ 11

Author(s):

Arash Kheradmand ◽

Majid Taati ◽

Homayoon Babaei

Keyword(s):

Plasma Membrane ◽

Treatment Group ◽

Sperm Motility ◽

Sperm Quality ◽

Sperm Concentration ◽

Membrane Integrity ◽

Chronic Administration ◽

Control Group ◽

Plasma Membrane Integrity ◽

Significant Difference

AbstractAlthough ghrelin acts as a modulator of feeding behavior and energy metabolism in the central nervous system, recent studies have implicated the peripheral actions of ghrelin in reproductive tissues. Here, we investigated the effects of chronic administration of ghrelin on the motility, plasma membrane integrity and concentration of rat spermatozoa. 45-d male Wistar rats were scheduled for the study and were divided into control and treatment groups. In the treatment group, 1 nmol of ghrelin was administered as sc injection for 10 consecutive days or vehicle (physiological saline) to the control rats. Sperm collection was achieved by killing of the rats on days 15, 25 and 50 after first injection. Total sperm motility and forward progressive movement did not exhibit significant difference during the experiment, although, there was a tendency for greater motion rate on d 15 and 25 in the treated rats compared to the control group. Plasma membrane integrity (HOS-reacted spermatozoa) was significantly higher in the treated animals, especially on day 15 as well as day 25, because of possible antioxidant properties of ghrelin. This value was statistically higher on day 15 than that of day 25 (P <0.05). Likewise, there was a significant correlation between the FPM (P <0.0001, r = 0.79) and TSM (P <0.01, r = 0.52) with the HOS test percentage in the treatment group. It was not observed statistically difference in the sperm concentration between groups during all of the experimental days. In conclusion, chronic administration of ghrelin (similar to induced by energy deficiency such as fasting) increased the integrity of sperm membrane, however, the sperm motility and concentration did not display any alterations.

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Effects of cryopreservation on semen quality and the expression of sperm membrane hexose transporters in the spermatozoa of Iberian pigs

Reproduction ◽

10.1530/rep-07-0118 ◽

2007 ◽

Vol 134 (1) ◽

pp. 111-121 ◽

Cited By ~ 32

Author(s):

S Sancho ◽

I Casas ◽

H Ekwall ◽

F Saravia ◽

H Rodriguez-Martinez ◽

...

Keyword(s):

Electron Microscopy ◽

Plasma Membrane ◽

Glucose Transporter ◽

Semen Quality ◽

Sperm Quality ◽

Membrane Integrity ◽

Membrane Receptors ◽

Plasma Membrane Integrity ◽

Sugar Transporters ◽

Hexose Transporters

This study evaluated the effects of cooling, freezing and thawing on the plasma membrane integrity, kinetics and expression of two sugar transporters glucose transporter-3 and -5 (GLUT-3 and GLUT-5) in spermatozoa from Iberian boars. Semen samples were collected twice weekly from eight young, fertile Iberian boars of the ‘Entrepelado’ and ‘Lampiño’ breeds. The samples were suspended in a commercial extender and refrigerated to 17 °C for transport to the laboratory (step A), where they were further extended with a lactose–egg yolk-based extender and chilled to 5 °C (step B) prior to freezing in the presence of glycerol (3%). Spermatozoa were assessed for plasma membrane integrity and sperm motility at each of the steps, including post-thaw (step C). Aliquots were also prepared for immunocytochemical localisation of the sugar transporters (fixed and thin smears for transmission and scanning electron microscopy levels respectively) and for SDS–PAGE electrophoresis and subsequent western blotting, using the same antibodies (rabbit anti-GLUT-3 and anti-GLUT-5 polyclonal antibodies). The results showed lower percentages of progressively motile spermatozoa at step C in both breeds, while the percentage of live spermatozoa was significantly lower only in the ‘Entrepelado’ breed. The results obtained from electron microscopy clearly showed that Iberian boar spermatozoa expressed the hexose transporters, GLUT-3 and GLUT-5. The pattern of expression, in terms of location and concentration, was characteristic in each case but, in the case of isoform GLUT-5, it remained constant during the different steps of freezing–thawing protocol. These results indicate that cryopreservation affects the status of sperm cells of Iberian boars by altering the distribution of some membrane receptors and decreasing the percentage values of parameters linked to sperm quality.

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Effect of platelet activating factor (PAF) supplementation in semen extender on viability and ATP content of cryopreserved canine spermatozoa

Polish Journal of Veterinary Sciences ◽

10.2478/v10181-010-0019-y ◽

2010 ◽

Vol 13 (4) ◽

pp. 571-579 ◽

Cited By ~ 5

Author(s):

W. Kordan ◽

M. Lecewicz ◽

R. Strzeżek ◽

A. Dziekońska ◽

L. Fraser

Keyword(s):

Plasma Membrane ◽

Sperm Motility ◽

Mitochondrial Function ◽

Sperm Quality ◽

Membrane Integrity ◽

Platelet Activating Factor ◽

Computer Assisted ◽

Atp Content ◽

Plasma Membrane Integrity ◽

Semen Extender

Effect of platelet activating factor (PAF) supplementation in semen extender on viability and ATP content of cryopreserved canine spermatozoa The aim of this study was to investigate the effect of platelet activating factor (PAF) on the quality characteristics of cryopreserved canine spermatozoa. Cryopreserved semen of 5 mixed-breed dogs was treated with different concentrations of exogenous PAF (1 × 10-3M, 1 × 10-4M, 1 × 10-5M and 1 × 10-6M) and examined at different time intervals (0, 30, 60 and 120 min). Cryopreserved semen treated without PAF was used as the control. Sperm quality was evaluated for motility (computer-assisted semen analysis, CASA), mitochondrial function (JC-1/PI assay) and plasma membrane integrity (SYBR-14/PI assay and Hoechst 33258). Also, ATP content of spermatozoa was determined using a bioluminescence assay. Treatment of cryopreserved semen with 1 × 10-3 M PAF at 120 min of incubation resulted in significantly higher total sperm motility compared with the control. It was observed that PAF-improved total sperm motility was concurrent with enhanced sperm motility patterns after treatment of cryopreserved semen. Treatment of cryopreserved semen with PAF did not improve either sperm mitochondrial function or plasma membrane integrity, as monitored by different fluorescent membrane markers. Furthermore, ATP content of cryopreserved spermatozoa was significantly higher when PAF was used at a concentration of 1 × 10-3 M compared with the control and other PAF treatments, regardless of the incubation time. The findings of this study indicated that treatment with 1 × 10-3 M PAF at 120 min of incubation rendered better quality of cryopreserved canine semen, which was associated with improved sperm motility parameters and ATP content. It can be suggested that exogenous PAF addition is beneficial as a supplement for canine semen extender used for.

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Efficiency of Commercial Egg Yolk-Free and Egg Yolk-Supplemented Tris-Based Extenders for Dromedary Camel Semen Cryopreservation

Animals ◽

10.3390/ani9110999 ◽

2019 ◽

Vol 9 (11) ◽

pp. 999 ◽

Cited By ~ 5

Author(s):

Ayman Abdel-Aziz Swelum ◽

Islam M. Saadeldin ◽

Hani Ba-Awadh ◽

Mohsen G. Al-Mutary ◽

Abdullah F. Moumen ◽

...

Keyword(s):

Plasma Membrane ◽

Sperm Motility ◽

Sperm Quality ◽

Membrane Integrity ◽

Egg Yolk ◽

Dna Integrity ◽

Computer Assisted ◽

Dromedary Camel ◽

Plasma Membrane Integrity ◽

Semen Cryopreservation

This study compared the efficiency of commercial egg yolk-free (AndroMed, OPTIXcell) and egg yolk-supplemented (Triladyl, Steridyl) Tris-based extenders for semen cryopreservation in seven adult dromedary camels. The camel-specific extender SHOTOR was used as control. The collected semen samples were evaluated and diluted with SHOTOR, Triladyl, Steridyl, AndroMed, or OPTIXcell. The diluted semen was gradually cooled and equilibrated for two hours before liquid nitrogen freezing. Semen was evaluated prior to freezing and after freeze-thawing cycles for motility, kinetics, vitality, abnormality, plasma membrane integrity, and DNA fragmentation using computer-assisted sperm analysis. In pre-freezing evaluation, progressive sperm motility was higher in SHOTOR-diluted samples (21.54 ± 1.83) than in samples diluted with Steridyl, OPTIXcell, or AndroMed (15.76 ± 1.80, 17.43 ± 1.10, and 13.27 ± 1.07, respectively). Moreover, Triladyl and SHOTOR resulted in significantly (p < 0.05) better sperm vitality and DNA integrity than all other diluents, but Triladyl resulted in a significantly (p < 0.05) better plasma membrane integrity (87.77 ± 0.31) than SHOTOR (85.48 ± 0.58). In the post-thawing evaluation, Triladyl led to significantly (p < 0.05) higher sperm motility (38.63 ± 0.81%; p < 0.05) when compared to SHOTOR, Steridyl or AndroMed (35.09 ± 1.341%, 34.4 ± 0.84%, and 31.99 ± 1.48%, respectively), with OPTIXcell being the least efficient (28.39 ± 0.86%). Progressive sperm motility was the highest when using Triladyl. Post-thawing curvilinear, straight line and average path sperm velocities were highest with Triladyl and lowest with AndroMed. Triladyl led to the highest linearity coefficient and straightness sperm coefficient, while SHOTOR to the highest DNA and plasma membrane integrity. OPTIXcell and AndroMed resulted in poor post-thawing sperm vitality, while Steridyl was less efficient than Triladyl. The highest rate of sperm abnormalities was recorded with OPTIXcell and the lowest with SHOTOR or Triladyl. In conclusion, SHOTOR, Triladyl, Steridyl, AndroMed, and OPTIXcell can all be used for camel semen cryopreservation; however, SHOTOR and Triladyl provided the best post-thawing sperm quality. Based on our findings, Triladyl is the best commercially available extender for dromedary camel semen cryopreservation to date.

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68 DIFFERENCE IN THAWING CURVE OF STALLION FROZEN SEMEN DILUTED IN 2 EXTENDERS

Reproduction Fertility and Development ◽

10.1071/rdv27n1ab68 ◽

2015 ◽

Vol 27 (1) ◽

pp. 127

Author(s):

C. P. Freitas-Dell'aqua ◽

C. Ramires Neto ◽

Y. F. R. Sancler-Silva ◽

P. M. Papa ◽

J. A. Dell'aqua ◽

...

Keyword(s):

Plasma Membrane ◽

Sperm Quality ◽

Membrane Integrity ◽

Freezing Process ◽

Freezing And Thawing ◽

Becton Dickinson ◽

Plasma Membrane Integrity ◽

Frozen Semen ◽

Group 2 ◽

Artificial Vagina

Commercial freeze extenders have different composition and ratio of cryoprotectors; freezing and thawing protocols are different for each extender. The aim of this experiment was to observe the effect of thawing curve in stallion frozen semen with 2 commercial extenders. Two ejaculates from each of 9 stallions of different breeds (Quarter Horses and Mangalarga Marchador) were used. Semen was collected using an artificial vagina, and the ejaculate was divided into 2 groups following the manufacture's protocol: group 1 (INRA), in which the semen was diluted 1 : 1 with the extender INRA 96TM (IMV, Paillette Crista, France) and group 2 (BC), in which the semen was diluted (1 : 1) with the extender Botu-SemenTM (Botupharma, Brazil). The samples of the 2 groups were centrifuged at 600 × g for 10 min, the supernatant was discarded, and the pellet was resuspended with INRA FreezeTM (group INRA, IMV) and with BotucrioTM (group BC, Botupharma) at the concentration of sperm 100 × 106 sperm mL–1. After this, the semen was packaged in 0.5-mL straws. For each group the freezing process was carried out according to the manufacturer's instructions. The straws were thawed in a water bath with 3 different thawing curves: 37°C for 30 s (37/30), 46°C for 20 s (46/20), and 75°C for 7 s (75/7) before analysis. The aim of these rates is to keep the semen in 37°C post-thaw. The sperm kinetic analysis was performed by computerized method (CASA, HTM-IVOS, IMV, USA) and the analysis of plasma membrane integrity by flow cytometer (BD LSR Fortessa, Becton Dickinson, Mountain View, CA, USA). Data of sperm kinetic and of plasma membrane integrity were compared among the 3 thawing curves for one extender using analysis of variance. Differences were considered significant at a probability level of 5%. No differences were observed in total motility (%, BC 37/30 = 72.8 ± 14.4; BC 46/20 = 70.0 ± 14.2; BC 75/7 = 70.3 ± 12.0 v. INRA 37/30 = 57.2 ± 19.1; INRA 46/20 = 50.0 ± 21.9; BC 75/7 = 58.8 ± 20.8), progressive motility (%, BC 37/30 = 36.9 ± 8.2; BC 46/20 = 34.4 ± 10.5; BC 75/7 = 33.6 ± 7.8 v. INRA 37/30 = 25.3 ± 12.7; INRA 46/20 = 21.9 ± 13.9; BC 75/7 = 28.9 ± 14.8), rapid sperm (%, BC 37/30 = 59.7 ± 16.4; BC 46/20 = 56.8 ± 17.1; BC 75/7 = 58.1 ± 14.9 v. INRA 37/30 = 38.3 ± 20.9; INRA 46/20 = 35.3 ± 22.9; BC 75/7 = 44.4 ± 23.8), and plasma membrane integrity (%, BC 37/30 = 49.1 ± 14.8; BC 46/20 = 43.1 ± 13.1; BC 75/7 = 46.7 ± 11.8 v. INRA 37/30 = 32.2 ± 10.7; INRA 46/20 = 29.6 ± 10.1; BC 75/7 = 37.4 ± 9.1) among the 3 thawing curves for INRA and BC groups. In this study, we can conclude there is no influence of the 3 tested thawing curves in sperm quality for stallion frozen semen with INRA Freeze and Botucrio extenders.

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Effects of the platelet-activating factor (PAF) on selected quality parameters of cryopreserved bull semen (AI) with reduced sperm motility

Polish Journal of Veterinary Sciences ◽

10.1515/pjvs-2016-0019 ◽

2016 ◽

Vol 19 (1) ◽

pp. 147-158 ◽

Cited By ~ 2

Author(s):

M. Lecewicz ◽

W. Kordan ◽

A. Majewska ◽

S. Kamiński ◽

A. Dziekońska ◽

...

Keyword(s):

Plasma Membrane ◽

Sperm Motility ◽

Sperm Quality ◽

Membrane Integrity ◽

Platelet Activating Factor ◽

Quality Parameters ◽

Sperm Viability ◽

Plasma Membrane Integrity ◽

Positive Effects ◽

Bull Semen

Abstract The aim of the study was to determine the effects of platelet-activating factor (PAF) on selected quality parameters of cryopreserved bull semen with reduced sperm motility used for artificial insemination. The aim of experiment 1 was to identify the optimal concentration of the phospholipid able to preserve sperm viability. Cryopreserved semen was treated with different PAF concentrations: 1×10-5M, 1×10-6M, 1×10-7M, 1×10-8M and 1×10-9M. The experiment demonstrated that PAF at concentration 1×10-9M increased most the sperm viability parameters (motility parameters, plasma membrane integrity and mitochondrial function) after 120 min of incubation of thawed semen at 37°C. Cryopreserved bull semen with reduced sperm motility (below 70%) was supplemented with PAF in a concentration of 1×10-9M. A statistically significant increase in sperm motility, percentage of linear motile spermatozoa and VSL value was observed after 120 min incubation of sperm with 1×10-9M PAF. Sperm supplementation with PAF also had positive effects on plasma membrane integrity and percentage of spermatozoa with preserved mitochondrial transmembrane potential, but the differences were not statistically significant. The results indicated positive effects of PAF supplementation at a concentration of 1×10-9M on the selected sperm quality parameters in cryopreserved bull semen with reduced motility.

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PENGARUH METODE PENCUCIAN SPERMATOZOA SAPI ACEH TERHADAP MOTILITAS, PERSENTASE HIDUP, DAN INTEGRITAS MEMBRAN PLASMA UTUH SPERMATOZOA (The Infuence of Aceh Bull Spermatozoa Washing Method on Spermatozoa Motility and Plasma Membrane Integrity of Intact Spermatozoa)

Jurnal Medika Veterinaria ◽

10.21157/j.med.vet..v9i2.3808 ◽

2015 ◽

Vol 9 (2) ◽

Author(s):

Listin Handayani ◽

Dasrul Dasrul ◽

Muslim Akmal ◽

Cut Nila Thasmi ◽

Hamdan Hamdan ◽

...

Keyword(s):

Plasma Membrane ◽

Semen Quality ◽

Sperm Quality ◽

Membrane Integrity ◽

Quality Parameters ◽

Control Group ◽

Plasma Membrane Integrity ◽

Spermatozoa Motility ◽

Sperm Washing ◽

Fresh Semen

This study aimed to determine the effect of sperm washing by swim up and centrifugation in isotonic medium on sperm quality of aceh bull. In this study, fresh semen from healthy male aceh bull aged 3-4 months was collected using artificial vagina. Immediately after semen collection, fresh semen quality was examined macroscopically and microscopically. Subsequently, sperm washing was performed by centrifugation and swim up in sperm washing medium. Group 1 (P0) as control group, cement washed with isotonic solution (andromed medium: saline solution) with ratio of 1:8. 2. Group 2 (P1), cement was separated by centrifugation method, group 3 (P2), all cement was separated by swim up method then examined the sperm quality sperm washing results. Each treatment was repeated 5 times. Quality parameters measured were the percentage of spermatozoa motility, sperm viability, and plasma membrane integrity intact spermatozoa. Data were analyzed with analysis of variance one-way pattern, followed by Duncan's multiple test. The results showed the mean ± SD percentage of sperm motility of each treatment group (P0; P1; P2) respectively amounted to 72.00±3.74, 66.40±4.77, and 73.60±3.29%. The percentage of viability was 72.00 ±3.74%, 66.40±2.88%, 71.80±2.17%. The percentage of plasma membrane integrity is intact spermatozoa was 68.20±1.79%, 57.20±3.77%, 69.00±2.00%. Results of this study showed that the percentage of motility, live spermatozoa and plasma membrane integrity intact after separation by swim-up method were significantly different (P <0.05) compared with no separation.Key words: spermatozoa quality, aceh bulls, centrifugation, swim up

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Factors affecting storage of Slovak native rabbit semen in the gene bank

Zygote ◽

10.1017/s0967199417000454 ◽

2017 ◽

Vol 25 (5) ◽

pp. 592-600

Author(s):

Barbora Kulíková ◽

Marta Oravcová ◽

Andrej Baláži ◽

Peter Supuka ◽

Peter Chrenek

Keyword(s):

Plasma Membrane ◽

Liquid Nitrogen ◽

Semen Quality ◽

Sperm Quality ◽

Objective Assessment ◽

Membrane Integrity ◽

Computer Assisted ◽

Quality Traits ◽

Plasma Membrane Integrity

SummaryIn this study, fresh and frozen–thawed semen of Nitra and Zobor rabbit breeds were evaluated for potential inter-breed or inter-male differences in sperm quality traits. Individual male semen from four rabbits of each breed were diluted (v:v; 1:1) in a freezing medium composed of a commercial diluent, 16% of dimethyl sulphoxide (DMSO), 4% of Ficoll 70 and 2% of sucrose and frozen in liquid nitrogen vapours before being plunged into liquid nitrogen. Different motility traits, viability and plasma membrane integrity of fresh and frozen–thawed semen were evaluated in vitro using computer-assisted sperm analysis and flow cytometry. To evaluate the sperm fertilization ability, artificial insemination of fresh and frozen–thawed sperm was performed. Our results showed the effect of breed (P ≤ 0.05) on frozen–thawed sperm viability and plasma membrane integrity. Moreover, individual variability in semen quality among the rabbits was revealed (0.31 to 0.71 among quality traits). Our results thereby confirmed that the cryopreservation procedure could not ensure comparable sperm post-thaw survival for different breeds or males. Nevertheless, correlations between numbers of fresh total motile and progressively moving sperm and several quality parameters measured post thawing were revealed. Therefore, we suggest that the objective assessment of fresh rabbit sperm motility may be an effective indicator of frozen–thawed semen quality. Consequently, regular semen assessment is required in order to preserve good-quality insemination doses from native breeds.

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