scholarly journals Effect of inbreeding depression on bull sperm quality and field fertility

2017 ◽  
Vol 29 (4) ◽  
pp. 712 ◽  
Author(s):  
Jesús Dorado ◽  
Rosa Morales Cid ◽  
Antonio Molina ◽  
Manuel Hidalgo ◽  
Julia Ariza ◽  
...  
Keyword(s):  
Inbreeding Depression ◽  
Sperm Morphology ◽  
Semen Quality ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Calving Interval ◽  
Bull Semen ◽  
Head Displacement ◽  
Highly Active ◽  
And Function

The present study investigated the effect of inbreeding depression on sperm quality using automated and objective methods and subsequent effects on beef bull field fertility. Individual inbreeding coefficient (F) values and field fertility data were determined using a dataset of AI bulls belonging to the Spanish Retinta Breeders Association (Asociación Nacional de Criadores de Ganado Vacuno Selecto de Raza Retinta (ANCRE)). Animals were clustered in two groups according to the F values as follows: (1) a high inbreeding group (HI; F ≥ 13.5%, mean 16.3); and (2) a non-inbreeding group (NI; F = 0%). In total, 17 different assessments were performed in both experimental groups, including evaluation of sperm morphology, acrosomal and DNA status, sperm plasma membrane integrity and function (hypo-osmotic swelling test), 10 kinetic parameters and the structure of sperm subpopulations. Sperm morphology, acrosomal and DNA status and osmotic tolerance were similar in both groups. Three velocity parameters (curvilinear velocity, straight line velocity and average path velocity) and the amplitude of lateral head displacement were higher in HI (P < 0.05). Cluster analysis of kinematic parameters revealed three different sperm subpopulations (sP1, sP2 and sP3), with the proportion of the sP1 population (highly active but non-progressive spermatozoa) being significantly (P < 0.05) higher in the HI group. Field fertility was assessed using two calving record datasets. In a smaller database including only bulls evaluated in the present study, there was a significant increase in the calving interval of cows sired with HI bulls. Conversely, in an extended genetic analysis of the ANCRE database, inbreeding only explained a small part of the variation in calving interval, and the results of regression analysis were not significant among bulls. The findings of the present study suggest that high inbreeding levels have a moderate effect on bull semen quality, with an increased percentage of highly active but non-progressive spermatozoa, but only when F values reached a certain threshold. This motility pattern could explain, in part, the higher calving interval produced by inbred bulls under field conditions.

scholarly journals Macro- and microelements in serum and seminal plasma as biomarkers for bull sperm cryotolerance

2021 ◽  
Vol 63 (1) ◽  
Author(s):  
Maja Zakošek Pipan ◽  
Petra Zrimšek ◽  
Breda Jakovac Strajn ◽  
Katarina Pavšič Vrtač ◽  
Tanja Knific ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Semen Quality ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Quality Characteristics ◽  
Freezing And Thawing ◽  
Additional Tool ◽  
Bull Semen ◽  
Bull Sperm ◽  
Fresh Semen

ABSTRACT Background Wide variation in fertility rates is observed when using frozen bull semen, even when the bulls have met quality standards for semen production. Therefore, a simple and reliable test to assess the freezing potential of bull semen based on the analysis of fresh semen or blood would be of great value. Attention is now turning to assessment of seminal plasma components such as proteins and elements. In the present study, the concentrations of macro- and microelements in fresh bull semen plasma and in serum and their correlation with quality characteristics of fresh semen and with semen quality after freezing and thawing were determined. Ejaculates were collected from 30 mature bulls, and semen volume, concentration, sperm motility, morphology, tail membrane integrity, plasma membrane permeability and DNA fragmentation were determined on the day of collection and after freezing and thawing. The concentrations of macroelements (Na, Mg, K and Ca) and microelements (Cu, Fe, Zn and Se) were determined in the seminal plasma and serum. The semen samples were classified into satisfactory and unsatisfactory groups according to the fresh semen quality. Results Zinc and Se levels measured in serum were associated with almost all fresh and frozen-thawed semen quality characteristics, while Fe levels were associated only with acrosomal defects in fresh semen. Zinc and Fe levels in fresh seminal plasma were associated with various quality characteristics of fresh and frozen-thawed semen, while Se level in fresh seminal plasma was not associated with any of the semen quality characteristics. Conclusions Microelements were shown to be useful as biomarkers involved in the analysis of bull sperm quality and could be used as an additional tool to predict bull semen quality after freezing and thawing. Our results confirm that the analysis of Zn and Se levels in serum and Zn, Cu and Fe levels in fresh seminal plasma can provide information to discriminate between bull semen samples with spermatozoa with high or low cryotolerance.

scholarly journals Effect of Supplementation of Polyvinylpyrrolidone in Extender on Buffalo Semen Parameters

2020 ◽  
Vol 53 (1) ◽  
Author(s):  
Bushra Ismail Khan ◽  
Shamim Akhter ◽  
Sanwal Aslam ◽  
Rabea Ejaz
Keyword(s):  
Sperm Motility ◽  
Semen Quality ◽  
Membrane Integrity ◽  
Bubalus Bubalis ◽  
Semen Parameters ◽  
Suction Pump ◽  
Bull Semen ◽  
Buffalo Semen ◽  
Live Sperm ◽  
And Control

The current study was planned to evaluate the supplementation of Polyvinylpyrrolidone in extender on cryopreservation of Nili-Ravi buffalo bull semen. The semen samples were collected from Nili-Ravi buffalo (Bubalus bubalis) bull kept at SPU Qadirabad, District Sahiwal, Pakistan. Qualifying semen ejaculates having motility >60%, volume >5-6ml and concentration >0.5 billion/ ml were diluted 50 × 106 motile sperm ml approximately at 37°C in Tris-citric acid extender supplemented with different concentrations of PVP (0.01, 0.05, 0.1mM). The extender without PVP was kept as control. Semen was stored at 4°C for a period of 2 h and kept at 4°C for 4h. Semen was filled in 0.5 ml French straws using suction pump at 4°C, plunged and stored in liquid nitrogen (-196°C). Semen straws were rewarmed at 37°C for 30 seconds and assessed for sperm motility, plasma membrane integrity (PMI), dead sperm percentage and the live sperm percentage. The data on the role of PVP on different parameters of semen quality were analyzed by using ANOVA and RCBD. Higher percentage (P< 0.05) of sperm motility (66.1±7.51 and 59.4±10.72) and PMI (72.9±5.39 and 75.7±6.5) was observed in extenders having 0.05 mM and 0.1mM PVP compared to extenders having 1.5mM PVP and control. The percentage acrosomal integrity was observed greater (P< 0.05) in extended semen containing 0.1mM (68.2±0.50) PVP compared to extenders having 0.01 and control.

scholarly journals Bacteriospermia and Sperm Quality of Cryopreserved Bull Semen Used in Artificial Insemination of Cows in South Wollo Zone, Ethiopia

2020 ◽  
Vol 2020 ◽  
pp. 1-11
Author(s):  
Abadi Amare Reda ◽  
Gizat Almaw ◽  
Solomon Abreha ◽  
Wedajo Tadeg ◽  
Belege Tadesse
Keyword(s):  
Bacterial Contamination ◽  
Semen Quality ◽  
Sperm Quality ◽  
Bacterial Load ◽  
Bacterial Count ◽  
Normal Morphology ◽  
Bull Semen ◽  
Significant Difference ◽  
Processing And Storage

The objectives of this trial were to estimate prevalence of bacteriospermia, to determine the bacterial load, and to isolate the types of bacteria as well as to assess the association between bacterial load and sperm quality traits in cryopreserved bull semen in field conditions in the South Wollo Zone. A total of 309 cryopreserved straws of semen from the Holstein Friesian (HF)-cross bull (n = 180 straws) and pure Jersey bull (n = 129 straws) were investigated. Bacteriological assessments of the presence of aerobic bacteria, estimation of bacterial count and bacterial isolation, as well as semen quality were performed. Aerobic bacterial contamination was prevalent in 38.8% of the semen straws. No significant difference in the prevalence of bacteriospermia was observed among bulls although the HF-cross bull had a higher prevalence (40.0%). But, significant difference in prevalence of bacteriospermia was found among semen ejaculates of the same bull. The risk of bacteriospermia in the HF-cross bull was higher (Odds ratio = 1.86, 95% CI = 0.168–20.26) compared to Jersey although not significant. Overall average bacterial load of 50.38 ± 16.29 colony-forming units (CFU)/ml (from nil to 1318.20 CFU/ml) was found. No significant difference in bacterial count among bulls and their ejaculates was observed. Moreover, correlation analysis revealed that the proportions of motility, live, and normal morphology were negatively influenced by an increase in the bacterial contamination of semen. In this study, three isolates of coagualse-negative Staphylococcus species and one isolate of Corynebacterium species were found. Average percentages of sperm motility (48.35 ± 1.23), live (66.08 ± 1.0), and normal morphology (80.62 ± 1.24) were observed. It was concluded that cryopreservation does not guarantee the quality of semen from bacterial contamination. Hence, meticulous care should be adopted to prevent contamination of semen by bacteria during collection, transportation, processing, and storage times.

8 THE USE OF ANNEXIN V MAGNETIC-ACTIVATED CELL SORTING TO SEPARATE APOPTOTIC SPERM FROM THE EJACULATE OF STALLIONS

2011 ◽  
Vol 23 (1) ◽  
pp. 110
Author(s):  
M. A. Coutinho da Silva ◽  
C. R. F. Pinto ◽  
J. M. Young ◽  
K. Cole
Keyword(s):  
Cell Sorting ◽  
Semen Quality ◽  
Sperm Quality ◽  
Cleveland Clinic ◽  
Membrane Integrity ◽  
Annexin V ◽  
Sperm Parameters ◽  
Control Group ◽  
Frozen Semen ◽  
Magnetic Activated Cell Sorting

Magnetic-activated cell sorting (MACS) has been used successfully in humans to remove apoptotic sperm from the ejaculate. Annexin V-conjugated microbeads recognise sperm with externalized phosphatidylserine, which is considered one of the features of apoptosis, and the labelled sperm is separated by MACS. The goals of the study were to determine if MACS can be used to separate apoptotic sperm from the ejaculate of stallions; and to determine if removal of apoptotic sperm improves the quality of stallion sperm. Our hypothesis was that MACS would improve semen quality by removing apoptotic sperm, resulting in samples with higher motility and viability. Two ejaculates from three different stallions of good fertility were used. Sperm were diluted with Tyrode’s albumin lactate pyruvate (TALP) and incubated with annexin V-conjugated microbeads for 15 min at 37°C. Control samples were incubated in the absence of annexin V microbeads. The suspension was then loaded into the separation column containing iron globes, which were fitted in a magnet (MiniMACS; Miltenyi Biotec Inc., Auburn, CA, USA). The effluent sample containing annexin-negative sperm was collected and then, the column was removed from the magnetic field and rinsed with TALP to collect the annexin-positive cells. Sperm viability, motility, morphology and caspase activation were determined in all three samples: control, annexin-negative, and annexin-positive. Data were evaluated by ANOVA and individual comparisons were performed by Tukey’s hsd test. Significance was set at P < 0.05 and data is presented as means ± SEM (Table 1). The main effect of stallion was significant only for sperm motility parameters. Sperm recovery rate following MACS was 46 ± 3%. In conclusion, the use of MACS was effective in removing apoptotic sperm from the ejaculate. The annexin-positive population displayed a higher proportion of sperm with activated caspases and lower membrane integrity and motility. However, removal of apoptotic sperm from the ejaculate did not improve sperm parameters in the annexin-negative group compared to control group. In addition, sperm morphology was not affected by MACS. Further studies are necessary to determine if MACS could be used successfully to improve sperm quality from subfertile stallions and frozen semen. Table 1.Sperm parameters following annexin V MACS (mean ± SEM) The authors are thankful to Mark Williams at Miltenyi Biotec Inc. for providing supplies; and Dr Ashok Agarwal at The Center for Reproductive Medicine, Cleveland Clinic, for scientific input.

23 Sperm quality of Pure Spanish stallions is affected by inbreeding coefficient and age

2020 ◽  
Vol 32 (2) ◽  
pp. 137
Author(s):  
Y. Pirosanto ◽  
M. Valera ◽  
A. Molina ◽  
J. Dorado ◽  
S. Demyda-Peyrás
Keyword(s):  
Sperm Motility ◽  
Inbreeding Depression ◽  
Negative Correlation ◽  
Semen Quality ◽  
Sperm Quality ◽  
Sperm Concentration ◽  
Inbreeding Coefficient ◽  
Semen Parameters ◽  
Computer Assisted

Inbreeding depression, a genetic condition produced by the mating of close-related individuals, has been associated with a reduction of fertility in several species. However, a loss in sperm quality was also associated with age. In horses, the few existing reports have described a tendency of both parameters to produce a negative effect on sperm quality. However, those reports were performed using a subjective evaluation of sperm motility. In the present study, a total of 692 ejaculates from 86 Pure Spanish stallions (PRE), aged between 3 and 22 years, were evaluated using a computer-assisted methodology to determine the effect of inbreeding in four semen parameters: free-gel volume (V), sperm concentration (C, by haemocytometer), and total (TM) and progressive (PM) sperm motility (by Spermvision sperm class analyser; Minitube). The inbreeding coefficient (F) was estimated using 300 000 PRE pedigree records approximately (minimum pedigree depth, eight equivalent complete generations; range, between 1 and 30.1%). Stallion, age, ejaculate, and season of semen collection were the variables included in the statistical model (general linear model), with ejaculate and season being the variables with a major effect (by variance components analysis). Our results showed that sperm concentration (r=−0.18; P&lt;0.0001) and volume (to a lesser extent) were reduced with advancing age, both showing a major decline after 15 years of age. To the contrary, sperm motility was not affected by age of the stallion. We also found a negative correlation between the inbreeding coefficient and ejaculate volume (r=−0.14; P&lt;0.001), with a marked decrease seen when F was between 7 and 20%. Also, a negative correlation was observed in PM (r=−0.08; P&lt;0.05), although to a lower extent. Conversely, C and TM were not affected by inbreeding depression (P&gt;0.05). In conclusion, our results demonstrated that high levels of inbreeding can compromise severely the sperm quality of the PRE stallion, which, subsequently, may have a negative influence on fertility. Ongoing studies using genomic data will help to detect genetic variants associated with stallion semen quality and how it is influenced by inbreeding in specific genomic regions.

Relationship between semen quality and seminal plasma cholesterol, triacylglycerols and proteins in the domestic cat

2019 ◽  
Vol 22 (10) ◽  
pp. 882-889 ◽  
Author(s):  
María F García ◽  
Romina Nuñez Favre ◽  
María C Stornelli ◽  
Ramiro Rearte ◽  
María C García Mitacek ◽  
...  
Keyword(s):  
Seminal Plasma ◽  
Sperm Morphology ◽  
Semen Quality ◽  
Sperm Count ◽  
Sodium Dodecyl ◽  
Sperm Concentration ◽  
Membrane Integrity ◽  
Domestic Cat ◽  
Sds Page ◽  
The Relationship

Objectives The current study aimed to evaluate the relationship between specific seminal plasma components – cholesterol (CHOL), triacylglycerols (TAG) and total protein (PROT) concentrations – and semen quality in cats. A further aim was to determine the relationship between specific seminal protein bands and semen quality. Methods Thirteen toms, 2–5 years of age, were included. Semen collection was performed by electroejaculation every 4 weeks. Fifty-eight ejaculates were assessed for motility, velocity, volume, sperm concentration, total sperm count, viability, acrosome integrity, plasma membrane integrity and sperm morphology. Samples were divided into two groups: good semen quality (GSQ) and poor semen quality (PSQ). After evaluation, seminal plasma was separated from the sperm by centrifugation and stored at −20°C. CHOL, TAG and PROT concentrations were then assessed and seminal plasma protein profile was determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Results Seminal plasma CHOL and TAG concentrations, motility, velocity, sperm concentration, total sperm count and sperm morphology were significantly higher in GSQ cats compared with PSQ cats ( P <0.01). Moreover, seminal plasma SDS-PAGE analysis showed an identifiable extra band exclusively in the GSQ group. Conclusions and relevance Data obtained in this study showed that seminal plasma CHOL and TAG concentrations and specific protein bands could be used to improve semen evaluation in toms. In this sense, the 14 kDa protein band could be a valuable marker for semen quality in the cat and should be further investigated. However, more studies are necessary to determine its relationship with fertility.

Evaluation of maize grain and polyunsaturated fatty acid (PUFA) as energy sources for breeding rams based on hormonal, sperm functional parameters and fertility

2012 ◽  
Vol 24 (5) ◽  
pp. 669 ◽  
Author(s):  
Sellappan Selvaraju ◽  
Priyadarshini Raju ◽  
Somu Bala Nageswara Rao ◽  
Subbarao Raghavendra ◽  
Sumantha Nandi ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Polyunsaturated Fatty Acid ◽  
Semen Quality ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
In Vitro Fertilisation ◽  
Functional Parameters ◽  
Maize Grain ◽  
Pufa Group ◽  
Different Sources

The objective of the present study was to elucidate the effect of different sources of dietary energy (maize vs polyunsaturated fatty acid (PUFA) on semen functional parameters and fertility of adult rams. Eighteen adult rams were divided into two groups (maize and PUFA, n = 9). The main energy source for the rams in the maize group was coarsely ground maize grain, whereas in the PUFA group it was sunflower oil (rich in 18 : 2 linoleic acid, an omega-6 acid). The ration was fed for a minimum period of 60 days and thereafter semen was collected for evaluation. The proportion of progressive forward motility was significantly (P < 0.05) higher in the PUFA group compared with the maize group. Sperm lipid peroxidation as measured by malondialdehyde formation (µM per 1 × 109 spermatozoa) was significantly (P < 0.05) higher in the PUFA group compared with the maize group. When the semen was diluted with Tris–egg yolk–citrate buffer and incubated for 24 h at 4°C, the proportions of plasmalemma integrity, the sperm subpopulation positive for functional membrane and acrosomal integrities, and mitochondrial membrane potential were significantly (P < 0.05) higher in PUFA-fed than in maize-fed animals. The different sources of energy did not influence the serum and seminal plasma IGF-I levels. The cleavage rate (percentage) did not differ significantly between PUFA- (45.4 ± 4.91) and maize- (44.63 ± 6.8) fed animals. In conclusion, PUFA feeding influenced sperm quality by altering or stabilising membrane integrity. The present study indicates that PUFA may improve semen quality but did not improve in vitro fertilisation.

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

Reproduction ◽  
2007 ◽  
Vol 134 (1) ◽  
pp. 111-121 ◽  
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.

scholarly journals Efecto de la composición racial sobre la calidad espermática de verracos

2018 ◽  
Vol 29 (3) ◽  
pp. 485 ◽  
Author(s):  
Anthony Valverde-Abarca ◽  
Mónica Madrigal-Valverde ◽  
Marlen Camacho-Calvo ◽  
Adones Zambrana-Jiménez ◽  
Leonardo López
Keyword(s):  
Quality Evaluation ◽  
Semen Quality ◽  
Sperm Quality ◽  
Active Movement ◽  
Racial Groups ◽  
Swine Farms ◽  
Progressive Motility ◽  
Head Displacement ◽  
Straight Line ◽  
Lateral Head

The assessment of the semen quality is an essential tool to improve the reproductive indexes in swine farms. The motility is the most important parameter that used in the sperm quality evaluation because it is associated with the energy of the spermatozoon. The aim of this research was to determine the influence of breed composition on reproductive variables of sperm quality, kinetics and semen motility in boars. During 2016, 240 ejaculates were collected from 63 boars with a mean age of 24.4±10.9 months. Six racial groups were identified: Duroc (D), Yorkshire (Y), Landrace (L), F1 Pietrain*Duroc (PD) and two genetic lines (LA and LB). The breeds with the highest ejaculate volume presented a higher total number of spermatozoa (P<0.05). The effect of the breed was significant (P<0.05) on sperm kinetics variables except for amplitude of lateral head displacement (ALH, μm). The Landrace breed presented the highest (P<0.05) percentage of static sperm (29.30±1.57). The most relevant differences (P<0.05) for total motility (MTOT) and progressive motility (MP), were presented between L and PD with values of 70.71±1.57; 77.48±1.09 and 51.80±1.97; 59.85±1.37% respectively. Adult boars (≥18 months) had higher volumes of ejaculate and total number of sperm than boars in the intermediate and young ages, however, for velocities (μm/s): curvilinear (VCL), straight line (VSL) and average path (VAP), adult boars only were different (P<0.05) of the boars in-between ages. Four sperm subpopulations (SP) were identified, SP1 (46.83%) characterized by a moderate speed, but of very progressive motility, SP2 (14.78%) with active movement, but not progressive, SP3 (8.45%) with low speeds and progressive path and SP4 (29.94%) with quick movement, but without progressivity.

scholarly journals Effect of whole and clarified egg yolk based extenders on post-thaw semen quality in bulls.

2021 ◽  
Author(s):  
Prosper Kamusasa ◽  
Eddington Gororo ◽  
Fungayi Primrose Chatiza
Keyword(s):  
Semen Quality ◽  
Sperm Quality ◽  
Egg Yolk ◽  
Quality Parameters ◽  
Normal Morphology ◽  
Semen Cryopreservation ◽  
Bull Semen ◽  
Inclusion Level ◽  
Whole Egg

Abstract This study was conducted to evaluate the comparative cryoprotective effects of whole egg yolk and clarified egg yolk on post thaw sperm quality parameters and to determine the optimum clarified egg yolk inclusion level (10-20%) in semen extenders for Mashona bull semen cryopreservation. It was shown that there was a significant decrease in sperm quality variables following cryopreservation. Semen quality increased with the concentration of clarified egg yolk, indicating a positive relationship between egg yolk LDL concentration and maintenance of in vitro sperm quality. The 20% clarified egg yolk (CEY20) extender treatment gave post-thaw motility, viability and normal morphology values which were comparable to the control (20% whole egg yolk, WEY20). The 10% clarified egg yolk concentration gave the least post-thaw quality values and the greatest proportion of defective spermatozoa. This experiment found no advantage of replacing whole egg yolk with up to 15% clarified egg yolk in Mashona bull semen cryopreservation. However, 20% clarified and 20% whole egg yolk performed similarly in the maintenance of post-thaw sperm motility, viability and normal morphology.

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