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.

72 EFFECT OF SINGLE-LAYER CENTRIFUGATION WITH EQUIPURE™ ON MOTILITY KINEMATICS OF FROZEN - THAWED DONKEY SPERM

2013 ◽  
Vol 25 (1) ◽  
pp. 183 ◽  
Author(s):  
I. Ortiz ◽  
J. Dorado ◽  
D. Acha ◽  
L. Ramirez ◽  
M. Urbano ◽  
...  
Keyword(s):  
Sperm Quality ◽  
Single Layer ◽  
Bottom Layer ◽  
Computer Assisted ◽  
Kinematic Parameters ◽  
Sperm Analysis ◽  
Progressive Motility ◽  
Head Displacement ◽  
Straight Line ◽  
Computer Assisted Sperm Analysis

Single-layer centrifugation (SLC) with EquipureTM Bottom Layer has been used to enhance the quality of stallion semen samples; however, no studies have been performed on donkeys. The aim of this study was to determine if SLC with EquipureTM Bottom Layer improves kinematic parameters on frozen–thawed donkey sperm. Semen was collected from 4 Andalusian donkeys by artificial vagina. Three ejaculates from each donkey were centrifuged with EquiproTM, supernatant was removed, and pellet was re-extended in the freezing medium GentTM to a final concentration of 200 × 106 spermatozoa per milliliter. Sperm were slowly cooled to 5°C for 2 h, loaded in 0.5-mL plastic straws, and frozen in liquid-nitrogen vapors. After at least one week of storage, straws were thawed in a water bath at 37°C for 30 s. After thawing, semen samples were divided in 2 aliquots: aliquot 1 was used as such (control) and aliquot 2 was processed by SLC using EquipureTM Bottom Layer. Computer-assisted sperm analysis was performed, and sperm kinematics total motility (%), progressive motility (%), curvilinear velocity (VCL; µm s–1), velocity straight line (VSL; µm s–1), velocity average path (VAP; µm s–1), linearity (LIN; %), straightness (STR; %), wobble (WOB; %), lateral head displacement (ALH; µm), and beat cross frequency (BCF; Hz) were statistically compared using GLM model between frozen–thawed semen samples processed or not with EquipureTM. Results were expressed as mean ± standard error. Significant differences (P < 0.05) were found between SLC-selected and unselected semen for total motility (77.44 ± 5.83 v. 58.89 ± 6.07), progressive motility (76.88 ± 4.52 v. 56.59 ± 5.44), VCL (137.50 ± 0.75 v. 133.0 ± 0.99), LIN (69.43 ± 0.31 v. 68.23 ± 0.41), STR (78.45 ± 0.29 v. 76.90 ± 0.37), WOB (85.06 ± 0.18 v. 83.91 ± 0.26), ALH (2.76 ± 0.01 v. 2.44 ± 0.01), and BCF (9.13 ± 0.05 v. 8.53 ± 0.06), respectively. No significant differences were observed for VSL (102.89 ± 0.70 v. 104.32 ± 0.95) and VAP (123.21 ± 0.71 v. 121.50 ± 0.98). Most of the computer-assisted sperm analysis parameters used in the present study have been previously identified as reliable markers of sperm motility in relation to sperm quality and fertility. It has also been reported that VCL appears to be critical for the formation of the sperm reservoir and penetration of the zona pellucida. In addition, other variables improved in the SLC-selected samples have been described as measure of progressivity (LIN, STR) and spermatozoa vigor (BCF, ALH). These preliminary results suggest an additional option for improving sperm quality in donkey semen doses. In conclusion, SLC with EquipureTM can be used to enhance kinematic parameters on frozen–thawed donkey sperm.

12 COMPUTER-ASSISTED SPERM ANALYSIS OF FRESH EPIDIDYMAL CAT SPERMATOZOA AND THE IMPACT OF COOLED STORAGE (4°C) ON SPERM QUALITY

2008 ◽  
Vol 20 (1) ◽  
pp. 86
Author(s):  
M. Filliers ◽  
T. Rijsselaere ◽  
P. Bossaert ◽  
V. De Causmaecker ◽  
J. Dewulf ◽  
...  
Keyword(s):  
Reference Values ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Computer Assisted ◽  
Epididymal Sperm ◽  
Sperm Analysis ◽  
Computer Assisted Sperm Analysis ◽  
The Impact ◽  
Motility Parameters

Feline epididymal sperm is commonly used for in vitro fertilization. It also yields the opportunity to conserve genetic material from valuable males that suddenly die. Epididymal sperm quality parameters vary considerably among laboratories, implicating the need for objective evaluation methods. The aim of the present study was to describe reference values of computer-assisted sperm analysis (CASA) parameters of fresh epididymal cat sperm and to assess the effect of prolonged cooled storage (4�C) on various sample characteristics. Epididymides obtained from tomcats after routine orchiectomy (2–4 pairs/replicate) were sliced to release spermatozoa. The sperm suspension was placed on a 2-layer gradient and, after centrifugation, the sperm pellet was recovered. In Experiment 1 (20 replicates), sperm motility parameters were assessed immediately after retrieval (T0) using the Hamilton Thorne analyzer Ceros 12.1 (HTR; Hamilton Thorne Biosciences, Beverly, MA, USA). In Experiment 2, fresh (T0) sperm samples (4 replicates) were evaluated for motility parameters (HTR), acrosomal status (FITC-Pisum sativum agglutinin staining), morphology (eosin/nigrosin (E/N) staining), and membrane integrity (E/N and SYBR�-14-propidium iodide staining; Molecular Probes, Inc., Eugene, OR, USA). After addition (1:2) of a Tris-glucose-citrate diluent containing 20% egg yolk, samples were cooled and reassessed on Days 1 (T1), 3 (T3), 5 (T5), 7 (T7), and 10 (T10). Results were analyzed in a mixed linear model, with replicate as random factor and time as fixed effect (S-PLUS 7.0; Insightful Corp., Seattle, WA, USA). Results of Experiment 1 were as follows (mean � SD): motility (MOT): 80.8% � 23.5; progressive motility (PMOT): 69.9% � 23.2; velocity average pathway (VAP): 98.7 µm s–1 � 24.2; velocity straight line (VSL): 89.3 µm s–1 � 25.4; velocity curved line (VCL): 134.8 µm s–1 � 31.9; amplitude lateral head (ALH): 4.3 µm � 2.0; beat cross frequency (BCF): 34.6 Hz � 7.0; and straightness (STR): 89.6% � 6.6. In Experiment 2, MOT, PMOT, VAP, VSL, VCL, BCF, and the percentage of normal spermatozoa showed a decrease over time (P < 0.05) compared to fresh samples, starting from T1, T3, T5, T7, T5, T3, and T1, respectively. In contrast, STR, ALH, membrane integrity, and the percentage of acrosome-intact spermatozoa were not affected (P > 0.05) by cooled storage. To summarize, we have presented a set of reference values for CASA-parameters of fresh, epididymal cat spermatozoa. Cooled storage impaired most motility parameters and lowered the percentage of normal spermatozoa, but did not influence membrane integrity or acrosomal status. The effect of cooled storage on DNA fragmentation of sperm and its subsequent influence on in vitro embryo development require further investigation.

scholarly journals Computer-Assisted Sperm Analysis of Freezable and Nonfreezable Mithun (Bos frontalis) Semen

2014 ◽  
Vol 2014 ◽  
pp. 1-6 ◽  
Author(s):  
P. Perumal ◽  
S. K. Srivastava ◽  
S. K. Ghosh ◽  
K. K. Baruah
Keyword(s):  
Objective Analysis ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Progressive Motility ◽  
Head Displacement ◽  
Straight Line ◽  
Computer Assisted Sperm Analysis ◽  
Lateral Head ◽  
Motility Parameters

The present study was undertaken to assess the motility and velocity parameters of sperm of freezable and nonfreezable ejaculates by computer-assisted sperm analyser (CASA) such as Hamilton-Thorne Semen Analyser IVOS 11 in mithun semen. Fifty ejaculates (twenty-five ejaculates each for freezable and nonfreezable semen ejaculates) were collected from ten matured mithun bulls. CASA parameters, motility parameters such as forward progressive motility (FPM) (%), nonprogressive motility (NPM) (%), total motility (TM) (%), and static sperms (SM) (%); velocity parameters such as curvilinear velocity (VCL) (μm/sec), straight line velocity (VSL) (μm/sec), average path velocity (VAP) (μm/sec), linearity (LIN) (%), straightness (STR) (%), wobble (WOB) (%), amplitude of lateral head displacement (ALH) (μm), and beat/cross-frequency (BCF) (Hz) were measured by CASA analyser. The result revealed that these parameters varied significantly (P<0.05) between the freezable and nonfreezable ejaculates and freezable ejaculates have significantly (P<0.05) higher value than nonfreezable ejaculates. It was concluded that most of the CASA parameters were significantly lower in nonfreezable ejaculates than in freezable ejaculates in mithun and confirmed that the CASA was effective for a quick and objective analysis of motility and velocity parameters in mithun semen.

scholarly journals Evaluation of outer dense fiber-1 and -2 protein expression in asthenozoospermic infertile men

2015 ◽  
Vol 24 (2) ◽  
pp. 79-83
Author(s):  
Silvia W. Lestari ◽  
Dwi A. Pujianto ◽  
Purnomo Soeharso ◽  
Evelyn Loanda
Keyword(s):  
Protein Expression ◽  
Sperm Motility ◽  
Computer Assisted ◽  
Fibrous Sheath ◽  
Sperm Analysis ◽  
Straight Line ◽  
The Mean ◽  
Computer Assisted Sperm Analysis ◽  
And Control ◽  
Infertile Patients

Background: Most of male infertility are caused by defect in sperm motility (asthenozoospermia). The molecular mechanism of low sperm motility in asthenozoospermic patients has not been fully understood. Sperm motility is strongly related to the axoneme structure which is composed of microtubules and supported by outer dense fiber (ODF) and fibrous sheath (FS) protein. The objective of this study was to characterize the ODF (ODF1 and ODF2) expression in asthenozoospermic infertile male and control normozoospermic fertile male.Methods: Asthenozoospermic samples (n=18) were collected from infertile patients at Andrology Lab, Cipto Mangunkusumo Hospital Jakarta and control were taken from normozoospermic fertile donor (n=18). After motility analyses by computer-assisted sperm analysis (CASA), semen were divided into two parts, for Western blot and for immunocytochemistry analysis. Antibody against ODF1 and ODF2 protein were used in both analyses.Results: Analysis of ODF1 protein expression showed bands with molecular weight of ~30 kDa and ODF2 ~85 kDa. The mean band intensity of ODF1 and ODF2 protein were lower in the asthenozoospermic group (AG) compared to normozoospermic group (NG). Moreover, both ODF proteins were less intense and less localized in the AG than NG. Sperm motility was lower in AG, compared to control NG, i.e. average path velocity (VAP) = 32.07 ± 7.03 vs 37.58 ± 8.73 µm/s, p = 0.455; straight line velocity (VSL) = 24.17 ± 6.90 vs 27.61 ± 4.50 µm/s, p = 0.317 and curvilinear velocity (VCL) = 45.68 ± 7.91 vs 55.55 ± 16.40 µm/s, p = 0.099.Conclusion: There is down-regulation of ODF1 and ODF2 protein expression and less-compact localization in AG sperm compared to the NG. These changes might have caused disturbances in the sperm motility as observed in this study.

scholarly journals Optimization of CASA-Mot Analysis of Donkey Sperm: Optimum Frame Rate and Values of Kinematic Variables for Different Counting Chamber and Fields

Animals ◽  
2020 ◽  
Vol 10 (11) ◽  
pp. 1993
Author(s):  
Sabrina Gacem ◽  
Jaime Catalán ◽  
Anthony Valverde ◽  
Carles Soler ◽  
Jordi Miró
Keyword(s):  
High Resolution ◽  
Sperm Motility ◽  
Frame Rate ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Head Displacement ◽  
Straight Line ◽  
Counting Chamber ◽  
Computer Assisted Sperm Analysis ◽  
Lateral Head

In order to optimize the donkey sperm motility analysis by the CASA (Computer Assisted Sperm Analysis)-Mot system, twelve ejaculates were collected from six jackasses. Capillary loaded chamber (CLC), ISAS®D4C depths 10 and 20 µm, ISAS®D4C Leja 20 and drop displacement chamber (DDC), Spermtrack® (Spk) depths 10 and 20 µm were used. Sperm kinematic variables were evaluated using each chamber and a high-resolution camera capable of capturing a maximum of 500 frames/second (fps). The optimum frame rate (OFR) (defined according to curvilinear velocity—VCL) was dependent on chamber type. The highest OFR obtained was 278.46 fps by Spk20. Values for VCL, straight-line velocity (VSL), straightness (STR), amplitude of lateral head displacement (ALH) and beat cross frequency (BCF) were high in DDC and 10 µm depth. In both DDC 10 and 20 µm, the sperm velocities (VCL, VSL, VAP) and ALH values decreased significantly from the centre to the edges, while Wobble and BCF increased. No defined behavior was observed along the CLC. However, all the kinematic variables had a higher value in a highly concentrated sample, in both chamber types. In conclusion, analyzing a minimum of nine fields at 250 fps from the centre to the edges in Spk10 chamber using a dilution of 30 × 106 sperm/mL offers the best choice for donkey computerised sperm motility analysis.

16 COMPUTER-ASSISTED SPERM ANALYSIS SPERM PARAMETERS OF BOS TAURUS CRIOLLOS BULL SEMEN v. THOSE OF OTHER BOS TAURUS BREEDS IN ARGENTINA

2008 ◽  
Vol 20 (1) ◽  
pp. 88
Author(s):  
M. G. Lüssenhoff ◽  
G. Larraburu ◽  
R. Cavia ◽  
A. Garcia Guerra ◽  
G. M. Brogliatti
Keyword(s):  
Sperm Motility ◽  
Sperm Morphology ◽  
Bos Taurus ◽  
Hybrid Vigor ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Bull Semen ◽  
Significant Difference ◽  
Computer Assisted Sperm Analysis ◽  
Motility Parameters

Criollos is the oldest cattle breed in America and the world at large. The origin dates back to the first cattle brought by C. Columbus. They are well known for their toughness and longevity. Their genetic variability is another advantage to be taken into account in crossbreeding because it ensures high hybrid vigor. The objectives of the present study were to determine computer-assisted sperm analysis (CASA) motility parameters and sperm morphology of Criollos bull semen (brought from Parque Nacional Los Glaciares, Patagonia, Argentina) v. other Bos taurus semen. A total of 29 ejaculates of different adult bulls (Angus and Hereford) and 25 ejaculates of Criollos bulls were evaluated in an artificial insemination center between December 2004 and December 2005. Semen collection was done by electroejaculation (EE) in all breeds. Fresh semen was diluted in a semi-defined semen extender (Andromed; Minitüb, Tiefenbach, Germany) and frozen in an automatic freezer (Digicool, IMV, Paillette Crista, France). Stain morphology was done using eosin/nigrosin, and evaluated in an optic microscope with phase contrast. Parameters of volume, total concentration (CONC), motility %, motile progressive sperm %, velocity average path (VAP, μm s–1), velocity straight line (VSL, μm s–1), curvilinear velocity (VCL, μm s–1), and linearity (LIN, %) were determined by CASA (HTM-ceros 12.1, Berkley, CA, USA). The results obtained were analyzed statistically with a one-way ANOVA test and are summarized in Table 1. Results from the sperm motility analysis indicate that there were no significant differences (P > 0.05) in CONC, motility %, motile progressive %, or VSL between Criollos and other B. taurus bull semen. VAP and VCL rates were significantly higher in Criollos bull semen than in other B. taurus semen, and LIN was lower. Also, there was a significant difference in volume collected between both breeds. Regarding sperm morphology, Criollos bull semen is at the maximum limit acceptable for head defects (detached heads: 15%). These results suggest that Criollos bull semen is not different from other Bos taurus semen; however, it does show different velocity and linearity rates. Table 1. CASA sperm motility parameters for semen from Criollos v. other Bos taurus bulls This research was supported by Centro Genetico Bovino Eolia S.A.

Motility of Fasciola hepatica miracidia assessed with a computer-assisted sperm analyser

2014 ◽  
Vol 89 (4) ◽  
pp. 453-457 ◽  
Author(s):  
A. Villa-Mancera ◽  
A. Reynoso-Palomar ◽  
J. Olivares-Pérez ◽  
S. Ortega-Vargas ◽  
I. Cruz-Mendoza ◽  
...  
Keyword(s):  
Fasciola Hepatica ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Straight Line ◽  
Different Temperatures ◽  
Motion Parameters ◽  
Computer Assisted Sperm Analysis ◽  
Incubation Temperatures ◽  
Hatching Temperature ◽  
Motility Parameters

AbstractThe motility parameters of Fasciola hepatica miracidia were assessed at different temperatures and times post-hatching using computer-assisted sperm analysis. Eggs were incubated at 22°C or 25°C for 14 days. Five motion parameters were evaluated at different incubation temperatures up to 10 h post-hatching. No differences were observed in the percentage that hatched after incubation at the two different temperatures. However, the straight-line velocity of miracidia following incubation at 22°C was significantly different from that observed at 25°C (P< 0.01). All miracidium motion parameters at different post-hatching temperatures showed an overall decrease at the end of the experiment. Those miracidia hatching from eggs incubated at 25°C had a higher velocity of 1673.3 μm/s compared with 1553.3 μm/s at 22°C. Velocity parameters increased as the post-hatching temperature increased from 22°C to 37°C.

Different computer-assisted sperm analysis (CASA) systems highly influence sperm motility parameters

2013 ◽  
Vol 80 (7) ◽  
pp. 758-765 ◽  
Author(s):  
S. Boryshpolets ◽  
R.K. Kowalski ◽  
G.J. Dietrich ◽  
B. Dzyuba ◽  
A. Ciereszko
Keyword(s):  
Sperm Motility ◽  
Computer Assisted ◽  
Sperm Analysis ◽  
Computer Assisted Sperm Analysis ◽  
Motility Parameters

scholarly journals Post-Thaw Sperm Quality and Functionality in the Autochthonous Pig Breed Gochu Asturcelta

Animals ◽  
2021 ◽  
Vol 11 (7) ◽  
pp. 1885
Author(s):  
José Néstor Caamaño ◽  
Carolina Tamargo ◽  
Inmaculada Parrilla ◽  
Felipe Martínez-Pastor ◽  
Lorena Padilla ◽  
...  
Keyword(s):  
Genetic Resource ◽  
Sperm Quality ◽  
Genetic Material ◽  
Mitochondrial Activity ◽  
Computer Assisted ◽  
Kinematic Parameters ◽  
Linear Mixed Effects ◽  
Sperm Analysis ◽  
Computer Assisted Sperm Analysis ◽  
Analysis System

Genetic resource banks (GRB) preserve the genetic material of endangered, valuable individuals or genetically relevant breeds. Semen cryopreservation is a crucial technique to reach these goals. Thus, we aimed to assess the sperm parameters of semen doses from the native pig breed Gochu Asturcelta stored at the GRB of Principado de Asturias (GRB-PA, Gijón, Spain), focusing on intrinsic and extrinsic (boar, season) factors. Two straws per boar (n = 18, 8–71 months of age) were thawed, pooled, and assessed after 30 and 150 min at 37 °C by CASA (computer-assisted sperm analysis system; motility and kinematic parameters) and flow cytometry (viability, acrosomal status, mitochondrial activity, apoptosis, reactive oxygen species, and chromatin status). The effects of age, incubation, and season on post-thawing quality were determined using linear mixed-effects models. Parameters were on the range for commercial boar breeds, with chromatin status (SCSA: fragmentation and immaturity) being excellent. Incubation decreased sperm quality and functionality. The boar age did not have a significant effect (p > 0.05), but the between-boar variability was significant (p < 0.001). The season significantly affected many parameters (motility, kinematics, viability, acrosomal status, mitochondrial activity), especially after 150 min of incubation. In general, samples collected in spring and summer showed higher quality post-thawing, the lowest in winter. In conclusion, the sperm doses from the Gochu Asturcelta breed stored at the GRB-PA showed excellent chromatin status and acceptable characteristics after thawing. Therefore, boar and seasonal variability in this autochthonous breed could be relevant for cryobank management.

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