180 ASSESSMENT OF BULL SEMEN QUALITY LOADED IN NEW SensiTemp STRAWS USING SEMEN AND IN VITRO PRODUCTION TECHNOLOGIES

2016 ◽  
Vol 28 (2) ◽  
pp. 221
Author(s):  
D. Le Bourhis ◽  
S. Camugli ◽  
P. Salvetti ◽  
L. Schibler ◽  
E. Schmitt
Keyword(s):  
Sperm Motility ◽  
Semen Quality ◽  
Sperm Quality ◽  
Semen Analysis ◽  
Membrane Integrity ◽  
Computer Assisted ◽  
Cleavage Rate ◽  
Fluid Medium ◽  
And Control

SensiTemp, a new in vitro maturation (IMV) bull straw concept, presents the advantage of colour changing while the straw is thawed. The colour of frozen straws is blue and straws start to become white when the temperature reaches 33°C, with a complete change of colour at 37°C. The objective of this study is to assess sperm quality after thawing of semen frozen in SensiTemp from 2 bulls, by analysing, in experiment 1, sperm motility and membrane integrity using computer-assisted semen analysis (CASA) and flow cytometry (FC), and, in experiment 2, the in vitro embryo production (IVP) using IVP technologies [IVM, IVF, and in vitro culture (IVC)]. The ejaculates of 2 bulls, selected during preliminary experiments on high in vitro fertility, were harvested at CIA L’Aigle, France, and split ejaculates were frozen in experimental (SensiTemp) and conventional (control) straws. In experiment 1 after thawing semen from the 2 types of straws (5 pooled straws each; 2 replicates), motility was assessed using the IVOS CASA system (Hamilton Thorne Inc., Beverly, MA, USA) and membrane integrity was evaluated through FC with Cytosoft software (Millipore-Guava Technologies Inc., Hayward, CA, USA). In experiment 2, IVF was used to evaluate the non-toxicity of SensiTemp and control straws. Cumulus-oocyte complexes (COC; n = 1178; 4 replicates) collected from slaughterhouse ovaries were matured in IVM medium (TCM-199 with bicarbonate, Sigma-Aldrich, Saint Quentin Fallavier, France; 10 µg mL–1 FSH-LH, Reprobiol, Liège, Belgium; and 10% FCS, Thermo Fisher, Illkirch, France) for 22 h. After fertilization, presumptive zygotes of each group (SensiTemp and control for each bull) were cultured in synthetic oviduct fluid medium (SOF, Minitube, Tiefenbach, Germany) with 1% estrous cow serum (ECS) and 0.6% BSA (Sigma-Aldrich, France) up to 8 days. All cultures were conducted at 38.5C in 5% CO2, and 5% O2. The cleavage and blastocysts rates were evaluated on Days 3 and 7, respectively, for each group. Embryo quality was recorded on Day 7 according to the IETS evaluation. Data from each bull were analysed separately using the chi-squared test (P < 0.05). In experiment 1, neither sperm motility from bull 1 (61.2 and 60.5%) and bull 2 (66.2 and 66.5%) nor membrane integrity from bull 1 (58.6 and 52.2%) and bull 2 (61.0 and 61.9%) were different between SensiTemp and control, respectively. Results from experiment 2 showed no difference (P > 0.05) in cleavage rate between SensiTemp and control for the 2 bulls: 92.1 and 91.7% for bull 1 and 94.2 and 94.6% for bull 2 respectively. The blastocysts rate on Day 7 did not differ (P > 0.05) among groups (47.5, 47.1 and 51.3, 50.4% for SensiTemp and control bull 1 and bull 2, respectively) nor the quality of embryos retrieved in the different groups: 25.4, 23.3, and 30.8, 29.6% in grade 1 embryo for SensiTemp and control bull 1 and bull 2, respectively. Those results demonstrate, in vitro, that the new SensiTemp straws were non-toxic and did not affect the semen quality after thawing nor did the SensiTemp straws affect the ability of sperm cells to fertilize oocytes and produce 8-day-old embryos.

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

Zygote ◽  
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.

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.

Cryopreservation of collared peccary (Pecari tajacu L., 1758) epididymal sperm using extenders based on Tris and powdered coconut water (ACP®-116c)

Zygote ◽  
2018 ◽  
Vol 26 (4) ◽  
pp. 301-307 ◽  
Author(s):  
José A. B. Bezerra ◽  
Andréia M. Silva ◽  
Patrícia C Sousa ◽  
Lívia B. Campos ◽  
Érica C. G. Praxedes ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Semen Analysis ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Coconut Water ◽  
Computer Assisted ◽  
Epididymal Sperm ◽  
Sperm Plasma Membrane ◽  
Pecari Tajacu ◽  
Functional Membrane

SummaryThe aim of this study was to establish a functional freezing–thawing protocol for epididymal sperm of collared peccaries (Pecari tajacu L., 1758) by comparing different extenders. The epididymal sperm from 12 sexually mature males was recovered by retrograde flushing using Tris-based or coconut water-based (ACP®-116c) extenders. After initial evaluation, samples were diluted and frozen with the same extenders to which 20% egg yolk and 6% glycerol were added. After 2 weeks, thawing was performed at 37°C/60 s and sperm motility, vigour, morphology, functional membrane integrity, sperm viability, sperm plasma membrane integrity, and a computer-assisted semen analysis (CASA) were assessed. In addition, to evaluate the survival of frozen–thawed sperm, a thermal resistance test (TRT) was executed. Samples preserved using Tris were in better condition compared with those preserved using ACP®, showing higher values for most assessments performed, including CASA and the TRT (P<0.05). After determining Tris to be the better of the two extenders, additional samples were thawed using different thawing rates (37°C/60 s, 55°C/7 s, 70°C/8 s). Sperm thawed at 37°C/60 s had the greatest preservation (P<0.05) of viability (54.1 ± 5.9%) and functional membrane integrity (43.2 ± 5.4%), and had higher values for various CASA parameters. In conclusion, we suggest the use of a Tris-based extender added to egg yolk and glycerol for the cryopreservation of epididymal sperm obtained from collared peccaries. In order to achieve better post-thawing sperm quality, we suggest that samples should be thawed at 37°C/60 s.

105 Increasing fertility of interspecific hybrid males using biotechnological approaches

2021 ◽  
Vol 33 (2) ◽  
pp. 159
Author(s):  
A. Vetokh ◽  
A. Tadzhieva ◽  
B. Iolchiev ◽  
N. Volkova ◽  
V. Bagirov
Keyword(s):  
Dna Fragmentation ◽  
Interspecific Hybrid ◽  
Semen Quality ◽  
Sperm Quality ◽  
Semen Analysis ◽  
Differential Staining ◽  
Computer Assisted ◽  
Sperm Dna Fragmentation ◽  
Russian Science

The results of AI depend on many factors, with the quality of semen being one of the most important. Not all male hybrids can meet the requirements for semen quality, because they often have reduced fertility following cryopreservation. Thus, it is necessary to improve semen processing before use in AI. The aim of the study was to evaluate the effectiveness of using the “swim-up” flotation method to improve sperm quality of hybrid males of the Ovis genus. Semen from interspecific hybrid rams (1/4 Argali×3/4 Romanov, n=15; 1/8 Argali×7/8 Romanov, n=15) was freshly obtained, frozen–thawed, and processed by the swim-up method. Evaluation of sperm motility was determined using computer-assisted semen analysis. Statistical analysis was performed using SPSS vs.15.0 (ANOVA and t-test; SPSS Inc.). Semen was collected during the breeding season (October–December) via artificial vagina. Assessment of acrosome integrity was determined using differential staining with a Diachem diff-quick kit (NPF ABRIS+). The degree of sperm DNA fragmentation was determined using the acridine-orange test. The sperm freezing/thawing cycle was accompanied by sperm damage and an increase in the proportion of immobile sperm from 10 to 58%, with non-progressive movement increasing from 9 to 19.3%. The number of spermatozoa with abnormal morphology doubled, and the DNA fragmentation index increased from 16 to 26%. Use of the swim-up procedure allowed us to sort progressively motile spermatozoa. The content of progressively motile spermatozoa in the samples obtained from the supernatant was 86%, which was 2.3 times higher than in frozen–thawed sperm (P≤0,01). The obtained results show the effective use of the swim-up procedure to determine the quality of semen in hybrid rams. These studies were carried out with financial support from the Russian Science Foundation, grant No. 18-16-00079 and the Ministry of Science and Higher Education of the Russian Federation.

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.

137 Assessment of fertilizing ability of Merino ram semen cold stored up to 48h by heterologous IVF of bovine oocytes

2019 ◽  
Vol 31 (1) ◽  
pp. 194 ◽  
Author(s):  
D. A. Galarza ◽  
M. Ladrón de Guevara ◽  
P. Beltrán-Breña ◽  
M. J. Sánchez-Calabuig ◽  
A. López-Sebastián ◽  
...  
Keyword(s):  
Semen Analysis ◽  
Skim Milk ◽  
Semen Parameters ◽  
Computer Assisted ◽  
Cleavage Rate ◽  
Straight Line ◽  
Fertilizing Ability ◽  
Pronuclear Formation ◽  
Bovine Oocytes

The use of cold-stored ram semen has been applied in sheep AI programs, because it preserves its fertilizing ability similar to fresh. Besides, the heterologous IVF has been successfully employed to assess semen fertilizing ability in several species. Hence, we aimed to evaluate the fertilizing ability of ram semen cold stored up to 48h at 5°C by assessing heterologous IVF using bovine oocytes. Fifteen pools of 3 normospermic Merino ram (2-7 years) ejaculates were collected using artificial vagina, diluted to 200×106 spermatozoa mL−1 with ultra-heat-treatment-based extender (skim milk-6% egg yolk) and cold stored up to 48h. In vitro matured zona-intact bovine oocytes were subjected to heterologous IVF using fresh semen (FS, n=707), semen cold stored to 24h (CS24, n=832) or semen cold stored to 48h (CS48, n=611). In parallel, homologous IVF (control, n=1356) and parthenogenesis (parth control non-fertilized oocytes, n=334) were performed. Ram non-selected and selected (BoviPure, Nidacon International, Mölndal, Sweden) semen parameters were evaluated by computer-assisted semen analysis. Sperm-oocyte interaction was assessed at 2.5h post-insemination (hpi) by evaluating the number of bound spermatozoa, whereas penetration and polyspermy were evaluated after 12 hpi. Presumptive zygotes were fixed and stained with Hoechst 33342 at 18, 20, 22, 24 and 26 hpi to assess pronuclear formation using phase contrast and confocal microscopy. Cleavage rate was evaluated in all groups at 48 hpi. Data obtained from 5 replicates were analysed using one-way ANOVA. Data was expressed as mean±standard error of the mean. In terms of sperm storage time, non-selected semen showed a significant decrease (P&lt;0.05) for CS24 and CS48 compared with FS on progressive motility [SPM (%): 52.30±4.1 and 36.9±5.5v. 71.3±1.6] and straight-line velocity (mm s−1: 132.2±6.1 and 109.7±6.3v. 176.7±4.3), respectively. However, selected semen showed a decrease (P&lt;0.05) only for CS48 when compared with CS24 or FS on SPM (35.6±3.9v. 56.1±6.91 and 59.3±2.6) and straight-line velocity (83.5±4.4v. 105.3±6.5 and 110±2.0), respectively. No differences were observed between heterologous IVF groups in all parameters evaluated. Homologous IVF showed a higher percentage of penetration only when compared with heterologous FS group (44.4±6.8v.12.5±4.5%; P&lt;0.01). The polyspermy was higher in heterologous CS24 group when compared with homologous IVF (11.4±3.4v. 3.8±2.2; P&lt;0.05). The homologous IVF group, as expected, showed the higher percentage of pronuclear formation at 18 hpi compared with heterologous IVF with FS (67.3±5.8v. 35.2±5.6%), CS24 (72.1±4.5 v. 37.2±5.7%) and CS48 (63.0±6.0 v. 27.0±5.6%), respectively (P&lt;0.001). Likewise, cleavage rate was higher in homologous group compared with heterologous IVF and parthenogenetic groups for FS (78.3±2.6.8v. 46.3±3.2 and 7.0±2.3%), CS24 (78.4±2.6 v. 48.3±3.2 and 4.9±2.0%), and CS48 (78.4±3.3 v. 43.3±3.5 and 4.3±1.2%), respectively (P&lt;0.001). In conclusion, Merino ram semen cold stored up to 48h maintains its fertilization ability to the same extend as fresh and can be used for sheep crossbreeding programs.

scholarly journals 12PREDICTION OF BULL FERTILITY BY COMPUTER ASSISTED SEMEN ANALYSIS

2004 ◽  
Vol 16 (2) ◽  
pp. 128 ◽  
Author(s):  
S. Cseh ◽  
T. Polichronopoulos ◽  
L. Solti
Keyword(s):  
Sperm Motility ◽  
Semen Analysis ◽  
Computer Assisted ◽  
Reproductive Process ◽  
Return Rates ◽  
Frozen Semen ◽  
Chi Square Analysis ◽  
Fertilizing Capability ◽  
Weighted Values

Sperm motility is clearly essential for fertilization both in vivo and in vitro. Motility is necessary for successful sperm transport, a step that is bypassed with in vitro fertilization. Recently, increasing attention has been paid to the objective evaluation and characterization of sperm motility more than simply determining the total proportion of motile spermatozoa. The purpose of computerassisted semen analysis (CASA) is to provide values for sperm concentration and sperm motility more rapidly and accurately than those obtained with traditional semen analyses methods. The objective of our experiment was to investigate the effect of specific aspects of sperm movement, such as the velocity of progression and the actual pattern of movement, to the fertilizing capability of sperm. Frozen semen samples of 10 HF breeding bulls were used in the study. For the motility analyses, Medealab CASA system (Medealab, Germany, Ver. 4.1) was used, and the velocity parameter of VCL (curvalinear velocity, μms−1), VSL (straight line velocity, μms−1), and VAP (average path velocity, μms−1) were evaluated and compared with the Day 30 and 75 non−return rates (NR30 and NR75). For every sample, a total of 10 fields were examined for 8s using a disposable 20 micron capillary chamber (CellVision, USA) giving a total of 1165 to 2831 cells evaluated. Chi square analysis, analyses of variance and linear correlation coefficient was applied to the statistical evaluation and comparison of the results. Data are based on weighted values. From the same batch of the analyzed frozen semen, a total of 8099 females were inseminated in more than 100 farms with a total of 6590 animals being positive for pregnancy at Day 30 and 4525 animals at Day 75. Within the bulls, differences were found in the values of NR30 and NR75 (P&lt;0.05). Our data indicate very strong differences between the males’ NR30 and NR75 values (NR30: 65.6%±13.04 to 79.6%±11.17; P&lt;0.001 and NR75: 37.8%±10.38 to 58.3%±15.53; P&lt;0.001) reflecting the individual differences in the fertilizing capability of the males. All velocity parameters show very high correlation with strong significance both non−return rates but the best values belong to VAP (NR30 and NR75; P&lt;0.02). Our data indicate that the bulls with lower VCL (25.51±33.04 to 79.54±58.03), VSL (11.35±19.45 to 36.36±35.71), and VAP (12.67±19.06 to 41.75±34.45) values showed lower fertilization rates both at NR30 and NR75. Computer and video technologies have advanced rapidly in recent years; thus the capability and accuracy of the latest versions of CASA systems are considerably better and they give more information about the different motion characteristics of spermatozoa. Because of the vital role of sperm motility in the reproductive process, such systems will enable us to move into a new era of diagnostic andrology and predict the fertilizing capability of semen. Supported by NKFP-Grants 4/040/2001 and 4/031/2001.

Effect of Panax notoginseng Extracts on Inferior Sperm Motility in vitro

1999 ◽  
Vol 27 (01) ◽  
pp. 123-128 ◽  
Author(s):  
Jung-Chou Chen ◽  
Ming-Xiong Xu ◽  
Leih-Der Chen ◽  
Yan-Nian Chen ◽  
Tsan Hung Chiu
Keyword(s):  
Sperm Motility ◽  
Sperm Count ◽  
Semen Analysis ◽  
Panax Notoginseng ◽  
World Health ◽  
Computer Assisted ◽  
Butanol Extract ◽  
The World ◽  
Health Organization

The purpose of this study was to investigate the effects of Panax notoginseng extracts on inferior sperm motility in vitro. Semen samples were collected from 23 patients with sperm motility between 20% and 40%. The sperm count was over 20 × 106/ml in accordance with the World Health Organization standard. 1.0 mg/ml and 2.0 mg/ml of Panax notoginseng extracts including aqueous extract, n-butanol extract, and polysaccharide fraction on sperm motility and progression were evaluated by computer assisted semen analysis. The results demonstrated that sperm motility as well as progression on inferior sperm motility were enhanced at 1 hour and 2 hours after incubation with all three types of extracts.

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

2010 ◽  
Vol 13 (4) ◽  
pp. 571-579 ◽  
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.

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

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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