Study of bovine sperm motility in shear-thinning viscoelastic fluids

Extent and timing of alterations in structures and functions of sperm after its placement in the female reproductive tract are important for successful fertilization. To our knowledge, the few reports are available on the kinetics of alterations in bovine sperm structures and functions during pathway to their death. Therefore, the present study was conducted to determine the changes in motility, acrosome and plasma membrane asymmetry in fresh and frozen–thawed semen during incubation at 37°C over the period of 24 h. Semen was collected from 3 breeding beef bulls, pooled, and considered as one replicate (total replicates = 5). Each pooled semen sample was diluted in Tris-citric acid egg yolk glycerol extender (pH 6.8), cooled to +4°C over 90 min, and then cryopreserved by a programmable cell freezer. Fresh (pooled semen) and frozen–thawed semen were incubated at 37°C for 24 h. Each semen sample was evaluated for sperm motility with computer-assisted semen analysis and acrosomal integrity and plasma membrane asymmetry using fluorescein isothiocyanate-peanut agglutinin/propidium iodide and Annexin V/propidium iodide assays, respectively, at 0, 2, 4, 6, 12, and 24 h of incubation at 37°C, with a flow cytometer. Statistical analysis was conducted using PROC MIXED model in statistical analysis system as 2 (semen types) × 6 (times) factorial model, using time as repeated measure. Progressive motility was higher (P < 0.05) in fresh than in frozen–thawed semen until 6 h. Progressive motility declined (P < 0.05) below the threshold level (i.e. 30%) much later (12 h) in fresh as compared with frozen–thawed semen (2 h). However, acrosomal integrity and plasma membrane asymmetry deteriorated (P < 0.05) below threshold at the same time interval (2 h) in both fresh and frozen–thawed semen. Viable sperm (AN–/PI–) remained higher (P < 0.05) during the first 6 h in fresh than in frozen–thawed semen and declined (P < 0.05) below the threshold at 12 h in fresh and at 6 h in frozen–thawed semen. In fresh semen, the necrotic sperm (AN–/PI+) population increased (P < 0.05) over time and reached maximum (97%) at 24 h. In frozen–thawed semen, a mixed population of late apoptotic (53%) and necrotic (34%) sperm was found at 24 h. In conclusion, the alterations in sperm motility, acrosomes, plasma membrane integrity, and asymmetry were slower in fresh than in frozen–thawed semen. Fresh sperm followed necrosis and frozen–thawed sperm underwent necrosis and apoptosis-like pathways, respectively. This study was supported by the Canadian Commonwealth Scholarship Program by the Canadian Bureau for International Education (CBIE), and Agriculture and Agri-Food Canada.

Download Full-text

Effects of shear thinning on migration of neutrally buoyant particles in pressure driven flow of Newtonian and viscoelastic fluids

Journal of Non-Newtonian Fluid Mechanics ◽

10.1016/s0377-0257(99)00074-9 ◽

2000 ◽

Vol 90 (2-3) ◽

pp. 159-185 ◽

Cited By ~ 39

Author(s):

P.Y. Huang ◽

D.D. Joseph

Keyword(s):

Viscoelastic Fluids ◽

Shear Thinning ◽

Pressure Driven Flow ◽

Neutrally Buoyant ◽

Pressure Driven

Download Full-text

Numerical study of driven flows of shear thinning viscoelastic fluids in rectangular cavities

Journal of Non-Newtonian Fluid Mechanics ◽

10.1016/j.jnnfm.2016.01.009 ◽

2016 ◽

Vol 229 ◽

pp. 59-78 ◽

Cited By ~ 7

Author(s):

Swapnil Dalal ◽

Gaurav Tomar ◽

Pradip Dutta

Keyword(s):

Numerical Study ◽

Viscoelastic Fluids ◽

Shear Thinning

Download Full-text

Involvement of signaling pathways in bovine sperm motility, and effect of ergot alkaloids

In Vitro Cellular & Developmental Biology - Animal ◽

10.1007/s11626-009-9191-8 ◽

2009 ◽

Vol 45 (8) ◽

pp. 483-489 ◽

Cited By ~ 13

Author(s):

Hehai Wang ◽

Michael L. Looper ◽

Zelpha B. Johnson ◽

Rick W. Rorie ◽

Charles F. Rosenkrans

Keyword(s):

Signaling Pathways ◽

Sperm Motility ◽

Ergot Alkaloids ◽

Bovine Sperm

Download Full-text

113 Effect of antioxidants on motility and fertility of liquid-stored bovine sperm

Reproduction Fertility and Development ◽

10.1071/rdv32n2ab113 ◽

2020 ◽

Vol 32 (2) ◽

pp. 183

Author(s):

Y. Honkawa ◽

T. Fujikawa ◽

N. Miura ◽

C. Kubota

Keyword(s):

Embryo Development ◽

Sperm Motility ◽

Sperm Concentration ◽

Frozen Storage ◽

Egg Yolk ◽

P Value ◽

Motile Sperm ◽

Progressive Motility ◽

Bovine Sperm ◽

Liquid Storage

It is difficult to maintain sperm in liquid storage for a long time, compared with permanent frozen storage in liquid nitrogen. Antioxidants have been reported to improve the quality and fertility of liquid-stored semen. In this study, we investigated whether antioxidants can extend the motility and fertility of frozen-thawed sperm in liquid storage. Frozen-thawed semen from one Japanese black bull (one ejaculate) was diluted in Tris-citrate-fructose (TCF) diluent with 10% (v/v) egg yolk to a sperm concentration of 1×107 spermmL−1. The antioxidants β-mercaptoethanol (βMe) and glutathione (GSH) were added independently, at various concentrations (0.1, 0.5, 1, and 5mM) to sperm suspensions, and these preparations were compared with Control (no added antioxidant). Sperm suspensions were packaged in centrifuge tubes and placed at 17°C in air and monitored daily until sperm motility had stopped (up to 14 days). Sperm motility was analysed by the Sperm Motility Analysis System (SMAS; Ditect Co. Ltd), and the percentage of progressively motile sperm (straight-line velocity (VSL) of >25μm s−1; Grade A classified by WHO manual), compared with that recorded on Day 0 (100%), was determined each day. For evaluation of fertilizing ability, after incubation in liquid storage for 0, 3, 5, and 7 days, sperm were used for IVF with invitro-matured oocytes (30 oocytes per treatment, three replicates). Embryo development was recorded as the proportion of embryos that reached blastocyst by 8 days after IVF. Data for motility were analysed using one-way ANOVA with Tukey test, and embryo development using chi-squared test. A P-value<0.05 was considered statistically significant. At 7 days, the percentage of progressively motile sperm was significantly higher for 0.5, 1, and 5mM βMe than for Control (30.8%, 48.1%, and 50.3%, vs. 0%, respectively). Treatments with 1 and 5mM βMe maintained some sperm progressive motility for 14 days (9.5% and 14.5%). Treatment with GSH showed the same trend at 7 days (32.2%, 36.3%, and 13.7% for 0.5, 1, and 5mM, vs. 0% for Control); 1 and 5mM GSH maintained sperm progressive motility over 10 days (24.8% and 4.4%). In both antioxidant treatments, embryo development was achieved with sperm stored for up to 5 days (Day 0 vs. Day 5 for 0.1mM βMe: 17.6% vs. 13.8%; for 1.0mM GSH: 26.0% vs. 6.7%; for Control: 17.6% vs. 0%). In this study, antioxidants extended both motility and fertility of frozen-thawed bovine sperm in liquid storage. This result suggests the possibility of application to AI using liquid-stored bovine semen.

Download Full-text

Effects of Ionic Strength on Lateral Particle Migration in Shear-Thinning Xanthan Gum Solutions

Micromachines ◽

10.3390/mi10080535 ◽

2019 ◽

Vol 10 (8) ◽

pp. 535 ◽

Cited By ~ 2

Author(s):

Mira Cho ◽

Sun Ok Hong ◽

Seung Hak Lee ◽

Kyu Hyun ◽

Ju Min Kim

Keyword(s):

Ionic Strength ◽

Rheological Properties ◽

Viscoelastic Fluid ◽

Xanthan Gum ◽

Viscoelastic Fluids ◽

Shear Thinning ◽

Particle Migration ◽

Cell Counting ◽

Contour Length ◽

Wide Range

Viscoelastic fluids, including particulate systems, are found in various biological and industrial systems including blood flow, food, cosmetics, and electronic materials. Particles suspended in viscoelastic fluids such as polymer solutions migrate laterally, forming spatially segregated streams in pressure-driven flow. Viscoelastic particle migration was recently applied to microfluidic technologies including particle counting and sorting and the micromechanical measurement of living cells. Understanding the effects on equilibrium particle positions of rheological properties of suspending viscoelastic fluid is essential for designing microfluidic applications. It has been considered that the shear-thinning behavior of viscoelastic fluid is a critical factor in determining the equilibrium particle positions. This work presents the lateral particle migration in two different xanthan gum-based viscoelastic fluids with similar shear-thinning viscosities and the linear viscoelastic properties. The flexibility and contour length of the xanthan gum molecules were tuned by varying the ionic strength of the solvent. Particles suspended in flexible and short xanthan gum solution, dissolved at high ionic strength, migrated toward the corners in a square channel, whereas particles in the rigid and long xanthan gum solutions in deionized water migrated toward the centerline. This work suggests that the structural properties of polymer molecules play significant roles in determining the equilibrium positions in shear-thinning fluids, despite similar bulk rheological properties. The current results are expected to be used in a wide range of applications such as cell counting and sorting.

Download Full-text