scholarly journals Tetrazolium Salt WST-8 as a Novel and Reliable Chromogenic Indicator for the Assessment of Boar Semen Quality

Animals ◽  
2020 ◽  
Vol 10 (12) ◽  
pp. 2293
Author(s):  
Yu-Hsin Chen ◽  
Chean-Ping Wu ◽  
Hsiu-Lien Lin ◽  
Ren-Bao Liaw ◽  
Yung-Yu Lai ◽  
...  
Keyword(s):  
Cell Viability ◽  
Semen Quality ◽  
Sperm Quality ◽  
Reduction Rate ◽  
Sperm Parameters ◽  
Cell Counting ◽  
Computer Assisted ◽  
Tetrazolium Salt ◽  
Reliable Prediction ◽  
Boar Semen

A tetrazolium salt, 2-[2-methoxy-4-nitrophenyl]-3-[4-nitrophenyl]-5-[2,4-disulfophenyl]-2H-tetrazolium (WST-8), has been used widely to determine cell viability; however, its application in the field of reproduction is still limited due to this assay merely providing information regarding cell viability. The aim of this study was to correlate the WST-8 reduction rate with various sperm quality-related parameters (i.e., sperm viability, motility, progressive motility, acrosome integrity and mitochondria integrity) in order to provide a rapid, reliable and affordable assessment for boar semen quality evaluation. Using different ratios of active/damaged sperm cells, we first validated our sample preparations by standard flow cytometry and computer-assisted sperm analysis. Further analyses demonstrated that the most efficient experimental condition for obtaining a reliable prediction model was when sperm concentration reached 300 × 106 cells/mL with the semen/cell-counting kit-8 (CCK-8®) ratio of 200/10 and incubated time of 20 min. Under this set up, the WST-8 reduction rate (differences on optic density reading value, ΔOD at 450 nm) and sperm parameters were highly correlated (p < 0.01) for all sperm parameters evaluated. In the case of limited semen samples, a minimal semen concentration at 150 × 106 cells/mL with the semen/CCK-8® ratio of 200/20 and incubation time for 30 min could still provide reliable prediction of sperm parameters using the WST-8 assay. Our data provide strong evidence for the first time that the WST-8 assay could be used to evaluate boar semen quality with great potential to be applied to different mammalian species.

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.

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.

scholarly journals Efficacy of potassium permanganate and turmeric as antimicrobial agents on the bacterial load and quality of boar semen during preservation at 15 °C

2020 ◽  
Vol 90 (5) ◽  
pp. 443-451
Author(s):  
Nang Kanna Manpoong ◽  
Kutubuddin Ahmed ◽  
Dilip Kumar Bhattacharya ◽  
Dipak Kumar Sarma ◽  
Nekibuddin Ahmed ◽  
...  
Keyword(s):  
Antimicrobial Agents ◽  
Semen Quality ◽  
Sperm Quality ◽  
Bacterial Load ◽  
Conception Rate ◽  
E Coli ◽  
Bacillus Spp ◽  
Boar Semen ◽  
Conventional Antibiotics

The present study was conducted to evaluate the efficacy of different natural antimicrobials agents (KMnO4 and Turmeric) in comparison with conventional antibiotics against the bacterial load and in relation to the quality of boar semen in Modena extender for up to 120 hours of preservation at 15 °C. A total of 56 ejaculates, 14 from each of four Hampshire crossbred boars maintained within the ICAR-AICRP on Pigs, in Guwahati, Assam, India, were utilized in the study. Thirty-two ejaculates, 8 from each of four boars were used to study the effect of antimicrobial agents on semen quality during preservation at 15 °C in Modena extender. A total of 9 different bacterial types were identified from 46 bacterial isolates, obtained from 24 fresh semen samples viz. Staphylococcus aureus (24%), E. coli (22%), Bacillus spp. (13%), Citrobacter spp. (9%), Pseudomonas spp. (9%), Staphylococcus epidermidis (9%), Klebsiella spp. (6%), Streptococcus spp. (6%) and Proteus spp. (2%). The overall sensitivity of the recovered isolates to Gentamicin, Ampicillin, Enrofloxacin, Cloxacillin, Streptomycin, Penicillin, Amoxycilln, Ofloxacin and Tetracyclin were 89, 39, 37, 48, 74, 52, 56, 76 and 63% respectively. The mean sperm motility, intact acrosome, HOST-reacted spermatozoa and bacterial load differed significantly (P˂0.01) between antimicrobial agents (Gentamicin, KMnO4 and Turmeric) and preservation periods (0, 48, 72, 96 and 120 hours). Sperm quality based on Gentamicin was found to be best, followed by Turmeric and KMnO4 during preservation at 15 °C. The conception rate for the semen preserved for 0, 24, 48, 72, 96 and 120 hours of preservation was 83.33, 80.00, 75.00, 66.66, 66.66 and 50.00% respectively. In the present study, the preserved semen with ascending bacterial load containing Gentamicin did not affect the conception rate.

145 ProAKAP4 concentrations in semen as a predictive tool of bull fertility: A preliminary study

2020 ◽  
Vol 32 (2) ◽  
pp. 199 ◽  
Author(s):  
I. Ruelle ◽  
N. Seregeant ◽  
D. Bencharif ◽  
F. Charreaux ◽  
C. Thorin ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Sperm Quality ◽  
Intrauterine Insemination ◽  
Human Reproduction ◽  
Sperm Parameters ◽  
Enzyme Linked Immunosorbent Assay ◽  
Computer Assisted ◽  
Predictive Tool ◽  
Progressive Motility ◽  
Preliminary Study

Recently, ProAKAP4 has been described as a pertinent indicator of sperm quality in humans, pigs, and stallions. In knockout mouse models lacking AKAP4 expression, the male mice were infertile. As high proAKAP4 levels were significantly correlated with a lower proportion of abortions in intrauterine insemination settings in human reproduction, proAKAP4 could be considered a pertinent new sperm parameter for assessing embryo quality. Our main goal was to assess the proAKAP4 concentrations in Holstein bull semen for comparison with the motility sperm parameters and fertility outcomes in post-thawed conditions. Straws issued from 52 ejaculates from 13 bulls, retrospectively identified with known nonreturn rates (NRR) as a fertility indicator, were provided by Evolution XY. Expression of ProAKAP4 and AKAP4 was assessed using enzyme-linked immunosorbent assay, western blotting, flow cytometry, and microscopy methods. Using the Bull 4MID kit (4BioDx), striking variations in proAKAP4 concentrations were observed independently of the classic sperm parameters that were measured using computer-assisted semen analysis. A mean proAKAP4 concentration of 44.42ng per 10 million spermatozoa was obtained through all our series. Interestingly, the variations in proAKAP4 concentrations were positively correlated with progressive motility and with the linearity coefficient parameter. Furthermore, the post-thawed concentrations of proAKAP4 were significantly higher in bulls with a higher NRR in a field study of more than 190 000 AI. We then demonstrated for the first time a correlation between the semen concentration of proAKAP4 and NRR (P=0.05) in bulls. Threshold values of proAKAP4 were then determined, with good values being between 25 and 60ngmL−1. Below 25ngmL−1, the sperm were of poor quality. The proportion of functional spermatozoa (i.e. spermatozoa expressing proAKAP4 in ejaculates) was assessed using flow cytometry. We observed that the cell debris and dead spermatozoa were never immunolabeled with proAKAP4 antibodies. On testis tissue sections, proAKAP4 was expressed only from the spermatids stages up to the ejaculated spermatozoa, being influenced by external factors and reflecting good spermatogenesis. Our preliminary study highlighted the pertinence of proAKAP4 in assessing sperm quality in bulls. It could be interesting to further analyse the effect of proAKAP4 level of expression on capacitation and IVF. As high levels of proAKAP4 were significantly correlated with fertility rates and with progressive motility, proAKAP4 could be proposed as a predictive marker of bull fertility and could be further investigated to evaluate the quality of invitro-produced embryos.

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.

Effects of low-density lipoproteins extracted from different avian yolks on boar spermatozoa quality following freezing–thawing

Zygote ◽  
2012 ◽  
Vol 22 (2) ◽  
pp. 175-181 ◽  
Author(s):  
Peng Wang ◽  
Yan-Feng Wang ◽  
Chun-Wei Wang ◽  
Shu-Hai Bu ◽  
Jian-Hong Hu ◽  
...  
Keyword(s):  
Semen Quality ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Egg Yolk ◽  
Quality Parameters ◽  
Low Density Lipoproteins ◽  
Low Density ◽  
Boar Sperm ◽  
Boar Semen ◽  
Avian Egg

SummaryLow-density lipoproteins (LDL) is known to protect boar sperm during freezing–thawing, but little information is known about the effects of LDL extracted from different avian egg yolks on post-thaw boar semen quality. The purpose of this study was to compare and analyze the effects of LDL at various concentrations and different species on boar sperm quality after freezing–thawing. LDL extracted from the yolk of hen egg, duck egg, quail egg, pigeon egg or ostrich egg was added to the extender at the concentrations of 0.06, 0.07, 0.08, 0.09 and 0.1 g/ml, respectively, and their effects on frozen–thawed boar sperm quality were assessed. According to all measured parameters, the results showed that sperm motility, acrosome integrity and plasma membrane integrity were 43.20%, 52.57% and 48.13%, respectively, after being frozen–thawed with 0.09 g/ml LDL extracted from pigeon egg yolk. All these quality parameters were higher than that of other groups (P < 0.05). In conclusion, our results confirmed that LDL extracted from pigeon egg yolk had the best cryoprotective effects on frozen–thawed boar sperm among all of the groups supplemented with LDL from five kinds of avian egg in extender. The optimum concentration of LDL extracted from pigeon egg in boar semen freezing extender was 0.09 g/ml.

Colloid single-layer centrifugation improves post-thaw donkey (Equus asinus) sperm quality and is related to ejaculate freezability

2015 ◽  
Vol 27 (2) ◽  
pp. 332 ◽  
Author(s):  
I. Ortiz ◽  
J. Dorado ◽  
D. Acha ◽  
M. J. Gálvez ◽  
M. Urbano ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Single Layer ◽  
Quality Parameters ◽  
Sperm Parameters ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Equus Asinus ◽  
The Relationship ◽  
Fresh Semen

The aim of this study was to determine whether colloid single-layer centrifugation (SLC) improves post-thaw donkey sperm quality and if this potential enhancement is related to ejaculate freezability. Semen from Andalusian donkeys was frozen following a standard protocol. SLC was performed on frozen–thawed semen and post-thaw sperm parameters were compared with uncentrifuged samples. Sperm quality was estimated by integrating in a single value sperm motility (assessed by computer-assisted sperm analysis), morphology and viability (evaluated under brightfield or fluorescence microscopy). Sperm freezability was calculated as the relationship between sperm quality obtained before freezing and after thawing. Ejaculates were classified into low, medium and high freezability groups using the 25th and 75th percentiles as thresholds. All sperm parameters were significantly (P < 0.01) higher in SLC-selected samples in comparison to uncentrifuged frozen–thawed semen and several kinematic parameters were even higher than those obtained in fresh semen. The increment of sperm parameters after SLC selection was correlated with ejaculate freezability, obtaining the highest values after SLC in semen samples with low freezability. We concluded that, based on the sperm-quality parameters evaluated, SLC can be a suitable procedure to improve post-thaw sperm quality of cryopreserved donkey semen, in particular for those ejaculates with low freezability.

scholarly journals Beneficial Influence of Soybean Lecithin Nanoparticles on Rooster Frozen–Thawed Semen Quality and Fertility

Animals ◽  
2021 ◽  
Vol 11 (6) ◽  
pp. 1769
Author(s):  
Lingwei Sun ◽  
Mengqian He ◽  
Caifeng Wu ◽  
Shushan Zhang ◽  
Jianjun Dai ◽  
...  
Keyword(s):  
Plasma Membrane ◽  
Semen Quality ◽  
Antioxidant Activities ◽  
Soybean Lecithin ◽  
Sperm Quality ◽  
The Other ◽  
Control Group ◽  
Mitochondrial Activity ◽  
Computer Assisted ◽  
The Impact

The present study aimed to investigate the impact of different concentrations (0%, 0.5%, 1.0%, 1.5%, and 2.0%) of nano-soybean lecithin (SL) in the extender on sperm quality, sperm motion characteristics, and fertility outcomes of post-thawed rooster semen. Adult Ross broiler breeder roosters (n = 20) were subjected to semen collections twice a week for three weeks. At each collection, semen samples were pooled and allocated into five treatments corresponding to different nano-SL concentrations (control, SL0.5, SL1.0, SL1.5, and SL2.0). Sperm parameters, including motility (collected using a computer-assisted sperm analysis system), plasma membrane and acrosome integrities, and mitochondrial activity were assessed. Sperm malondialdehyde (MDA) and antioxidant activities (total antioxidant capacity (TAC); superoxide dismutase (SOD); glutathione peroxidase (GPx)) were evaluated. The fertility and hatchability obtained with frozen–thawed rooster semen supplemented with the optimum nano-SL concentration were assessed after artificial insemination. The results showed that the addition of 1% nano-SL into the extender led to a higher semen motility in roosters, improved plasma membrane and acrosome integrities, and higher mitochondrial activity of post-thawed rooster semen in comparison to controls (p < 0.05). The MDA levels in the SL0.5 and SL1.0 groups were lower than the other groups (p < 0.05). TAC activities in SL0.5, SL1.0, and SL1.5 groups were significantly higher than those in the other groups (p < 0.05). It was observed that the concentration of SOD was higher in the SL1.0 group than in the other groups (p < 0.05). The activity of GPx was not influenced in any of the cases (p > 0.05). Moreover, the percentages of fertility and hatchability in the SL1.0 group were higher (56.36% and 58.06%) than those in the control group (42.72% and 40.43%). In summary, the addition of nano-SL to the extenders enhanced the post-thawed semen quality and fertility of roosters by reducing the level of oxidative stress. The optimum nano-SL concentration was 1.0%. These results may be beneficial for improving the efficacy of semen cryopreservation procedures in poultry breeding.

30 Bull sperm kinetics after semen cryopreservation in extender containing propagermanium

2019 ◽  
Vol 31 (1) ◽  
pp. 141
Author(s):  
T. E. Cruz ◽  
A. Martins Jr ◽  
F. N. Marqui ◽  
D. G. Souza ◽  
T. I. H. Berton ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Membrane Lipid ◽  
Free Radical Scavenger ◽  
Sperm Parameters ◽  
Radical Scavenger ◽  
Computer Assisted ◽  
Kinetics Parameters ◽  
Significance Level ◽  
Freezing Medium ◽  
Bull Sperm

There is a negative association between a plentiful production of oxygen reactive species and spermatozoa kinetics parameters. Thus, antioxidants have been added to the freezing medium to improve sperm quality due to their protective effect against membrane lipid peroxidation. Propagermanium (GE132) is an organometallic compound that has never been used in freezing medium despite its known antioxidant effect as a free radical scavenger. This study aimed to investigate the effects of different concentrations of GE132 added in a commercial freezing medium (CFM) on frozen-thawed sperm motion. Nine ejaculates of 3 Nellore bulls (3 replicates), collected by an artificial vagina, were evaluated, pooled, and divided into groups D (semen was diluted and kept at 33°C for 30min before cooling) and C (semen was cooled immediately after dilution). Both groups were submitted to the same experimental treatment, as follows: addition of 0, 500, and 1000µg mL−1 of GE132 in a CFM resulting in subgroups D0, D500, D1000, C0, C500, and C1000. The sperm samples were diluted to a final concentration of 30×106 spermatozoa per straw (0.25mL) and then cooled at 4°C for 5h before freezing. Sperm samples were assessed using a computer assisted sperm analyser at 5 and 60min post-thawing for total motility (TM;%), progressive motility (%), curvilinear velocity (μm s−1), velocity straight line (μm s−1), velocity average path (μm s−1), amplitude of the lateral head displacement (ALH; μm), beat cross frequency (Hz), linearity (LIN;%), and straightness (STR;%). Data were analysed using the R software package version 3.4.4 (2018; https://www.r-project.org/). An ANOVA was applied to assess statistical differences, and Tukey’s test was used to determine differences among subgroups. A significance level of P&lt;0.05 was adopted. No significant differences (P&gt;0.05) were observed among subgroups for all sperm parameters except for TM, in which C0 presented higher (P&lt;0.05) value (68.72±3.36) than D0 (54.67±5.59), D5 (57.10±2.34), and C10 (54.20±2.73), with similar results between D10 (59.5±4.22) and C5 (59.52±4.64). There were significant differences within subgroups when comparing 5 and 60min post-thawing for TM, ALH, LIN, and STR. Total motility decreased 17.2 and 9.9% in C5 and C10, respectively. Similarly, values of ALH decreased 0.2, 0.4, and 0.2µm in D0, D5, and C5, respectively. However, the increase in LIN was 11% in D10, whereas the values for STR increased in D10 (10%), C5 (6.3%), and C10 (7.3%). The addition of GE132 to the CFM did not enhance all the sperm parameters after cryopreservation except for a slight improvement in ALH, LIN, and STR over time and TM among groups. The lack of additive effect could be due to the presence of antioxidants in the CFM; therefore, further investigation with fluorescent probes using flow cytometry and free-antioxidants freezing medium could lead to a new approach for bull sperm freezing. We acknowledge Tairana AI Station, Master Fertility, and Botupharma, Brazil.

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