Swim-up of tammar wallaby (Macropus eugenii) spermatozoa in Biggers, Whitter and Whittingham (BWW) medium: maximisation of sperm motility, minimisation of impairment of sperm metabolism and induction of sperm hyperactivation

2017 ◽  
Vol 29 (2) ◽  
pp. 345 ◽  
Author(s):  
Minjie Lin ◽  
Xiyi Zhang ◽  
Ray N. Murdoch ◽  
R. John Aitken
Keyword(s):  
Sperm Motility ◽  
First Record ◽  
Tammar Wallaby ◽  
The Other ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Rapid Induction ◽  
Marsupial Species ◽  
Accurate Quantification ◽  
Better Than

A variety of media were compared for their ability to sustain the motility of tammar wallaby spermatozoa over an 8-h period following swim–up from coagulated semen. The study demonstrated that a modified Tyrode’s solution, Biggers, Whitter and Whittingham medium (BWW) was significantly better than any of the other assessed media in supporting wallaby sperm motility. After 8 h of incubation in BWW, motility was maintained at 79.3 ± 9.3%, with 77.0 ± 10.4% rapid and 65.7 ± 8.7% progressively motile spermatozoa. By contrast, motility was <10% at the same 8-h time point in all of the other media assessed. After 2 h of incubation in BWW, tammar spermatozoa consumed more oxygen than their counterparts in PBS (52.0 ± 2.7 vs 75.0 ± 6.6 μL per 108 spermatozoa per 2 h; P < 0.001). Motility was not enhanced in any of these media by the addition of 5 mM N-acetyl-D-glucosamine, the major energy substrate in wallaby semen. However, addition of dibutyryl cAMP and pentoxifylline in BWW resulted in the extremely rapid induction of hyperactivated motility in the entire sperm population. This burst of hyperactivated motility was entirely dependent on calcium in BWW and significantly inhibited by calmidazolium, a calmodulin inhibitor. A set of computer-assisted sperm analysis parameters were identified that permitted the accurate quantification of hyperactivation rates in this species. This is the first comparative analysis of media for harvesting and incubating marsupial spermatozoa and the first record of hyperactivated motility in any marsupial species.

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

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

The objective of this study was to optimize bovine sperm storage for up to 20 h between semen collection and sex sorting followedby cryopreservation. Two successive ejaculates were obtained from mature dairy bulls (Holstein, n = 5; Jersey, n = 3) via artificial vagina. Treatments were then applied to the neat semen to which antibiotics were added as recommended by Certified Semen Services (Columbia, MO). Nothing further was added to the control samples until staining with Hoechst 33342 for sorting. For Treatment 1, semen was diluted 9:1 with a MOPS solution resulting in 24 mM MOPS and similarly, Treatment 2 resulted in 24 mM MOPS +2% egg yolk. A subsample of each treatment and control was sorted by flow cytometry shortly after collection, and sperm then were frozen following standard processing procedures. The other subsample was stored at 15-18°C and sorted 20 h after collection followed by cryopreservation. pH measurements were made before staining samples for sorting. Samples were evaluated post-thaw for subjective progressive and total sperm motility, by computer-assisted sperm analysis (CASA, Berkeley, CA, USA), and by flow cytometry for sperm viability using propidium iodide and SYBR-14. Treatment 1 performed better than the control (Table 1), while results for Treatment 2 were similar to the control. Second ejaculates were superior to first ejaculates. pH measurements showed that addition of MOPS kept the pH about 0.2 units higher than the control, but pH declined similarly over time in all groups. While responses for the 20 h sort were numerically lower than the 0 h sort (P > 0.1), the majority of responses were acceptable for most, but not all bulls. In conclusion, storing sperm in 24 mM MOPS was beneficial. Surprisingly, 2% egg yolk negated the beneficial effect of MOPS, possibly due to increasing osmolarity by ∼15mOsM/kg due to pH adjustment. Addition of MOPS provided better results than the control for both the 0 h and 20 h sorts. Table 1.Main effect means of semen characteristics

17 Increased inbreeding levels negatively affect sperm kinetics and motility in Purebred Spanish horses

2021 ◽  
Vol 33 (2) ◽  
pp. 116
Author(s):  
Y. Pirosanto ◽  
A. Molina ◽  
M. Valera ◽  
J. Dorado ◽  
E. Terán ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Sperm Quality ◽  
Reproductive Traits ◽  
Study Groups ◽  
Inbreeding Coefficient ◽  
Computer Assisted ◽  
Effective Population ◽  
Mean Velocity ◽  
Sperm Analysis ◽  
Head Displacement

Reproductive performance is one of the key factors in livestock production. It is well known that reproductive traits are influenced by several genetic factors, such as the increase of individual inbreeding levels, which are associated with changes in sperm motility and shape in several species. In horses, the increase in inbreeding is a common problem because of the reduction in effective population size and the increase in selection intensity observed in several breeds. However, studies assessing the effect of high levels of inbreeding on the sperm quality of stallions are scarce. In the present study, we aimed to determine the effect of increased inbreeding levels and age on the sperm motility patterns of Purebred Spanish horses (PRE). We performed kinetic characterisation of 557 sperm samples of 82 PRE stallions aged between 3 and16 years, using computer-assisted sperm analysis (Androvision™, Minitube). We evaluated 5 parameters in 6 different fields per sample: curved line velocity (VCL, µm/s), velocity average path (VAP, µm/s), velocity straight line (VSL, µm/s), amplitude of lateral head displacement (ALH, µm), and beat-cross frequency (BCF, Hz). We determined the pedigree-based inbreeding coefficient (Fped) based on ∼300,000 PRE pedigree records to evaluate the inbreeding effect. Individuals were separated into 2 groups: highly inbred (n=339) and lowly inbred (n=218) according to an F value of 12.5%. Differences between groups were analysed using a generalized linear model. The analysis did not show significant differences (P&gt;0.05) in the variables analysed with respect to the age of stallions. However, VAP, VCL, and AHL were lower in highly inbred than in lowly inbred animals (P&lt;0.05), suggesting less velocity and amplitude of head displacement. In the case of BCF, no significant differences (P&gt;0.05) were observed between the two study groups. In conclusion, age did not affect sperm quality parameters in the age group of stallions analysed. In addition, we demonstrated that high inbreeding coefficient reduced the mean velocity and trajectory pattern of spermatozoa in PRE.

scholarly journals Effects of chilled storage and pH of activating solution on different motility parameters in burbot (Lota lota) sperm

2018 ◽  
Vol 63 (No. 11) ◽  
pp. 429-434
Author(s):  
Zoltán Bokor ◽  
Balázs Csorbai ◽  
Levente Várkonyi ◽  
Zsolt Szári ◽  
Ferenc Fodor ◽  
...  
Keyword(s):  
Sperm Motility ◽  
Sperm Quality ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Chilled Storage ◽  
Straight Line ◽  
H 26 ◽  
Computer Assisted Sperm Analysis ◽  
Motility Parameters ◽  
Activating Solution

The effects of a simple saline solution prepared using two different pH (4.4 and 8.5) on sperm motility in burbot were investigated. Results were recorded during a 96-hour chilled storage (4°C) in 24-hour intervals. Measurements were focused on the detailed characteristics of motility using 12 parameters obtained from the Computer-assisted Sperm Analysis (CASA). Significantly higher progressive motility (pMOT), distance average path (DAP), distance curved line, distance straight line (DSL), average path velocity (VAP), curvilinear velocity, straight line velocity, and beat cross frequency (BCF) were observed with the activating solution buffered at pH 8.5 in comparison with pH 4.4. Already after 24 h a significant reduction was measured in pMOT (0 h: 49 ± 24%, 24 h: 12 ± 7%). Similar decreasing tendency was recorded only after 72 h in DAP (0 h: 26 ± 4 µm/s, 72 h: 19 ± 9 µm/s), DSL (0 h: 21 ± 5 µm/s, 72 h: 17 ± 8 µm/s), VAP (0 h: 59 ± 9 µm/s, 72 h: 43 ± 21 µm/s), and BCF (0 h: 28 ± 2 Hz, 72 h: 18 ± 10 Hz). The response of different investigated CASA parameters to different treatments varied in our experiments. According to our studies, numerous burbot sperm motility parameters are sensitive to chilled storage and to low pH of the activating solution. Our results could support the effective sperm quality assessment and successful artificial propagation process in burbot.

scholarly journals Effects of Astaxanthin on Miniature Pig Sperm Cryopreservation

2018 ◽  
Vol 2018 ◽  
pp. 1-9 ◽  
Author(s):  
Eunjoo Lee ◽  
Daeyoung Kim
Keyword(s):  
Sperm Motility ◽  
Semen Parameters ◽  
Control Group ◽  
Computer Assisted ◽  
Miniature Pig ◽  
Oxidation Stress ◽  
Sperm Analysis ◽  
Semen Parameter ◽  
Boar Sperm ◽  
Sperm Plasma Membrane

The purpose of this study is to evaluate the effects of astaxanthin added to freezing buffer on semen parameters, total sperm oxidation stress after postthawing of boar sperm, and lipid peroxidation (LPO) which is caused by reactive oxygen species (ROS) in sperm membrane. Varying concentrations of astaxanthin (0, 10, 50, 100, and 500 μM) were used in the freezing buffer during cryopreservation to protect the DNA of thawed miniature pig sperm. Semen parameter was measured using computer-assisted sperm analysis (CASA) for sperm motility, and then ROS rate and oxidative stress of boar sperm were determined using fluorescence-activated cell sorting (FACS). Sperm motility was higher (p<0.05) in the astaxanthin group than in the control group. Sperm motility and the number of progressive motile sperm were higher (p<0.05) in the astaxanthin 500 μM group than in the control group. In ROS evaluation, the astaxanthin group had lower intracellular O2 and H2O2 in viable sperm. Yo-Pro-I/HE and PI/H2DCFDA staining as revealed using flow cytometry was lower in astaxanthin groups than in the other groups. As a result, we found that astaxanthin could protect the sperm plasma membrane from free radicals and LPO during boar sperm postthawing.

Chlamydia trachomatis-induced death of human spermatozoa is caused primarily by lipopolysaccharide

2003 ◽  
Vol 52 (3) ◽  
pp. 193-200 ◽  
Author(s):  
S. Hosseinzadeh ◽  
A.A. Pacey ◽  
A. Eley
Keyword(s):  
Chlamydia Trachomatis ◽  
Sperm Motility ◽  
Polymyxin B ◽  
Probit Analysis ◽  
Human Spermatozoa ◽  
Similar Proportion ◽  
Computer Assisted ◽  
Osmotic Swelling ◽  
Potent Effect ◽  
Sperm Analysis

Elementary bodies (EBs) of Chlamydia trachomatis serovar E are more toxic to sperm than those from serovar LGV. In this study, lipopolysaccharide (LPS) was prepared from the EBs of both serovars and incubated with human spermatozoa at concentrations that matched the LPS concentration of EBs. The effects of EBs and LPS on sperm motility, viability and acrosomal status were then determined. Sperm motility was measured by computer-assisted sperm analysis and the hypo-osmotic swelling test was used to determine the proportion of dead cells. Acrosomal status was examined using a standard mAb assay. Over a 6 h incubation, LPS from both serovars resulted in a marked reduction in sperm motility (and a concomitant increase in the proportion of dead spermatozoa) in a manner similar to that seen in response to EBs of serovar E. In addition, when sperm were incubated with a range of doses of EBs and LPS, probit analysis revealed that the greater spermicidal effects of EBs from serovar E (when compared with serovar LGV) were not observed when sperm were incubated with LPS from the two serovars. This suggests that the more potent effect of EBs of serovar E cannot be explained entirely by differences in the composition of LPS. Interestingly, Escherichia coli LPS was required in doses 500 times more concentrated than chlamydial LPS in order to kill a similar proportion of sperm, suggesting that bacterial LPSs may differ in their spermicidal properties. However, that chlamydial LPS was spermicidal was demonstrated by the use of polymyxin B (a polycationic antibiotic known to neutralize LPS effects), confirming that the effects observed were primarily a result of LPS activity.

Application of Computer-assisted Sperm Analysis in Selecting the Suitable Solution for Common Carp,Cyprinus carpioL., Sperm Motility

2013 ◽  
Vol 44 (3) ◽  
pp. 466-472 ◽  
Author(s):  
Beata Irena Cejko ◽  
Beata Sarosiek ◽  
Radosław Kajetan Kowalski ◽  
Sławomir Krejszeff ◽  
Dariusz Kucharczyk
Keyword(s):  
Common Carp ◽  
Sperm Motility ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Suitable Solution ◽  
Computer Assisted Sperm Analysis

scholarly journals Nitric oxide stimulates human sperm motility via activation of the cyclic GMP/protein kinase G signaling pathway

Reproduction ◽  
2011 ◽  
Vol 141 (1) ◽  
pp. 47-54 ◽  
Author(s):  
Erica Miraglia ◽  
Federico De Angelis ◽  
Elena Gazzano ◽  
Hossain Hassanpour ◽  
Angela Bertagna ◽  
...  
Keyword(s):  
Nitric Oxide ◽  
Protein Kinases ◽  
Sperm Motility ◽  
Soluble Guanylate Cyclase ◽  
Human Sperm ◽  
Computer Assisted ◽  
No Donor ◽  
Sperm Analysis ◽  
Cgmp Analog ◽  
Dependent Protein

Nitric oxide (NO), a modulator of several physiological processes, is involved in different human sperm functions. We have investigated whether NO may stimulate the motility of human spermatozoa via activation of the soluble guanylate cyclase (sGC)/cGMP pathway. Sperm samples obtained by masturbation from 70 normozoospermic patients were processed by the swim-up technique. The kinetic parameters of the motile sperm-rich fractions were assessed by computer-assisted sperm analysis. After a 30–90 min incubation, the NO donor S-nitrosoglutathione (GSNO) exerted a significant enhancing effect on progressive motility (77, 78, and 78% vs 66, 65, and 62% of the control at the corresponding time), straight linear velocity (44, 49, and 48 μm/s vs 34, 35, and 35.5 μm/s), curvilinear velocity (81, 83, and 84 μm/s vs 68 μm/s), and average path velocity (52, 57, and 54 μm/s vs 40, 42, and 42 μm/s) at 5 μM but not at lower concentrations, and in parallel increased the synthesis of cGMP. A similar effect was obtained with the NO donor spermine NONOate after 30 and 60 min. The GSNO-induced effects on sperm motility were abolished by 1H-[1,2,4]oxadiazolo-[4,3-a]quinoxalin-1-one (a specific sGC inhibitor) and mimicked by 8-bromo-cGMP (8-Br-cGMP; a cell-permeating cGMP analog); the treatment with Rp-8-Br-cGMPS (an inhibitor of cGMP-dependent protein kinases) prevented both the GSNO- and the 8-Br-cGMP-induced responses. On the contrary, we did not observe any effect of the cGMP/PRKG1 (PKG) pathway modulators on the onset of hyperactivated sperm motility. Our results suggest that NO stimulates human sperm motility via the activation of sGC, the subsequent synthesis of cGMP, and the activation of cGMP-dependent protein kinases.

94 ADDITION OF REDUCED GLUTATHIONE TO THAWING MEDIUM IMPROVED THE SPERM MOTILITY AND REDUCED ROS GENERATION IN FROZEN OVINE AND CAPRINE SPERMATOZOA

2006 ◽  
Vol 18 (2) ◽  
pp. 155 ◽  
Author(s):  
J. C. Gardón ◽  
J. A. Rodriquez ◽  
J. Gadea
Keyword(s):  
Sperm Motility ◽  
Reduced Glutathione ◽  
Ovis Aries ◽  
Capra Hircus ◽  
Ros Generation ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Spermatozoa Motility ◽  
Curvilinear Trajectory ◽  
Motility Parameters

The processes of cooling and freezing/thawing produce physical and chemical stress on the sperm membrane, and this stress is associated with oxidative stress and reactive oxygen species (ROS) generation that further reduce sperm viability and fertilizing ability. It is known that the process of freezing is associated with a significant reduction of the intracellular reduced glutathione (GSH) content. The aim of these experiments was to investigate the effects of addition of GSH to thawing extenders on motility parameters and ROS generation in frozen-thawed ovine and caprine spermatozoa. Frozen spermatozoa from eight rams (Ovis aries) and eight bucks (Capra hircus) (generously provided by Ovigen, Zamora, Spain) were thawed in a water bath at 37�C for 30 s and resuspended in sperm-TALP medium (Parrish et al. 1986 Theriogenology 25, 591-600) without (control) and with addition of 1 mM or 5 mM GSH. After 30 min of incubation at 37�C, sperm motility was evaluated using a computer-assisted sperm analysis (CASA) system (SCA, Microptic, Barcelona, Spain). The recorded parameters of motility were: % total, % progressive, curvilinear velocity, straight-line velocity, average path velocity, linearity of the curvilinear trajectory, straightness, amplitude of lateral head displacement, wobble of the curvilinear trajectory and beat cross frequency. Another set of sperm samples was incubated in the presence of (0.7 �M) 22,72-dichlorodihydrofluorescein diacetate (Gadea et al. 2005 J. Androl. 26, 396-404) to estimate production of ROS by flow cytometry. Data were analyzed by two-way ANOVA, considering the specific sperm treatment (GSH addition) and the males as the main variables. In ram frozen spermatozoa, all of the motility parameters were significantly improved when the medium was supplemented with GSH (P < 0.01) with even better results when 5 mM GSH was used. As an example, progresive motility increased from 31.16% (control) to 39.17 and 43.97%, respectively, for 1 and 5 mM GSH. Despite of the male effect detected (P < 0.01), all eight rams studied presented a similar pattern (interaction P > 0.05). The generation of ROS was significantly reduced when GSH was added (6.23a for control vs. 5.32b and 3.85c for 1 and 5 mM, respectively; P < 0.01). In buck frozen spermatozoa, % motility and progressive motility were significantly higher in GSH groups than in the control (P < 0.01), with no differences between 1 and 5 mM GSH. However, for the other motility parameters, the differences were not significant, which probably could be related to differences in the pattern shown by different animals (interaction of buck by treatment P < 0.05). ROS generation was significantly reduced when GSH was added (7.50a for control vs. 4.32b and 2.70b for 1 and 5 mM, respectively; P < 0.01). The addition of GSH to the thawing medium had a positive influence on the parameters studied in both species, increasing the motility patterns and reducing the ROS generation. In conclusion, we can assume that the addition of reduced glutathione to the thawing medium exerts a protective effect on spermatozoa functionality. This work was supported by AGL-2003-03144.

51 CHANGING ROOSTER SPERM MEMBRANES TO FACILITATE CRYOPRESERVATION

2012 ◽  
Vol 24 (1) ◽  
pp. 137 ◽  
Author(s):  
K. M. Tarvis ◽  
P. H. Purdy ◽  
J. K. Graham
Keyword(s):  
Liquid Nitrogen ◽  
Sperm Motility ◽  
Membrane Damage ◽  
Membrane Integrity ◽  
Computer Assisted ◽  
Membrane Composition ◽  
Sperm Analysis ◽  
Unsaturated Lipids ◽  
Multiple Comparison Test ◽  
Nitrogen Vapor

Cryopreservation damages rooster sperm membranes. Part of this damage is due to membrane transitioning from the fluid to the gel state as temperature is reduced. Some of this damage may be prevented by increasing membrane fluidity at low temperatures by incorporating cholesterol or unsaturated lipids into the membrane. Different concentrations of cholesterol-loaded cyclodextrins (CLC) and lipid-loaded cyclodextrins (LLC) containing 1,2-dilinoleoyl-sn-glycero-3-phosphocholine; 1,2-dilinoleoyl-rac-glycerol; and 1,2-dilinolenoyl-sn-glycero-3-phosphocholine were added to rooster sperm to determine if they improved cryopreservation. Osmotic stresses when cryoprotectants (CPA) are added to the cells before freezing and when the CPA are removed from cells after thawing also cause membrane damage. To minimize this damage, low molecular weight CPA with high membrane permeability were tested to determine their effectiveness for cryopreserving sperm. Rooster semen was collected from several birds, pooled and diluted to 800 million sperm mL–1 at 5°C in Lake's Low Temperature diluent (LLT). Sperm were treated with either LLC (0.25, 0.5, 1, 1.5, 2, 4 and 6 mg mL–1) or CLC (0.5, 1 and 2 mg mL–1) for 30 min. The sperm were diluted 1:1 with LLT containing 18% CPA, resulting in final CPA concentrations of 9%. The CPA tested were glycerol (G), methylacetamide (MA), dimethylformamide (DMF), methylformamide (MF) and ethylene glycol (EG). The sperm were frozen in liquid nitrogen vapor and stored in liquid nitrogen. Straws were thawed in 5°C water and sperm motility and membrane integrity analysed immediately. Sperm motility was measured using computer-assisted sperm analysis (CASA) and membrane integrity was analysed by flow cytometry using propidium iodide to detect cells with damaged membranes. Data were analysed by ANOVA and means separated using Student–Newman–Keuls multiple comparison test. Addition of LLC and CLC did improve sperm cryosurvival rates (P > 0.05). Using G as the CPA resulted in higher percentages of motile (54%) and viable (58%) sperm than MA (47 and 52%; P < 0.05), whereas DMF, EG and MF resulted in less than 45% motile cells (P < 0.05). In conclusion, altering sperm membrane composition using CLC and LLC did not improve post-thaw motility or viability in rooster sperm. Although MA did not protect the rooster sperm from cryodamage as effectively as G, future assays will need to determine the fertilizing capacity of sperm frozen using these CPA. We thank CSU-CVBMS for funding support.

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

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

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

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