Optimization of a continuous real-time computerized semen analysis system for ram sperm motility assessment, and evaluation of four methods of semen preparation

1996 ◽  
Vol 8 (2) ◽  
pp. 219 ◽  
Author(s):  
WV Holt ◽  
MJ Palomo
Keyword(s):  
Real Time ◽  
Semen Analysis ◽  
Computer Assisted ◽  
Assessment And Evaluation ◽  
Analysis System ◽  
Set Up ◽  
Assisted Analysis ◽  
Ram Sperm ◽  
Preparation Techniques

Sampling conditions that affect the biological validity of computer-assisted analysis of ram sperm motion were examined using a continuous real-time computerized semen analysis system (Hobson Sperm Tracker). Search radius (SR, 10 settings) and minimum track point (MTP, 10 settings) were varied factorially to evaluate their effects on the inclusion of sperm subpopulations within derived datasets. Low SR (< 12 microns) or high MTP values (> 26 frames) precluded measurements of rapidly moving cells, whereas high SR (> 17 microns) and low MTP settings (< 22 frames) led to erroneous tracking and poor data quality. Suitable settings for these set-up parameters were derived and tested for their biological consistency with four methods of preparing ram semen for computerized analysis. The preparation techniques tested were: centrifugation through sucrose-based Ficoll and Percoll media, a swim-up technique and simple dilution in Tyrode's media. The 'selective' Percoll and swim-up methods generated sperm populations with significantly higher linear velocities and a lower tendency to deviate from linear trajectories than from either the Ficoll method or dilution technique. Deleterious effects of centrifugation were evident, particularly on sperm survival in vitro over several hours. It is concluded that computer-assisted semen analysis provides useful information about the behaviour of ram spermatozoa in vitro, but the measurement conditions must be defined carefully.

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.

scholarly journals Asthenozoospermia in Mice with Targeted Deletion of the Sperm Mitochondrion-Associated Cysteine-Rich Protein (Smcp) Gene

2002 ◽  
Vol 22 (9) ◽  
pp. 3046-3052 ◽  
Author(s):  
Karim Nayernia ◽  
Ibrahim M. Adham ◽  
Elke Burkhardt-Göttges ◽  
Jürgen Neesen ◽  
Mandy Rieche ◽  
...  
Keyword(s):  
Semen Analysis ◽  
Computer Assisted ◽  
Microscopical Examination ◽  
Structural Protein ◽  
Wild Type ◽  
Vitro Fertilization ◽  
In Vivo Experiments ◽  
Cysteine Rich Protein

ABSTRACT The sperm mitochondria-associated cysteine-rich protein (SMCP) is a cysteine- and proline-rich structural protein that is closely associated with the keratinous capsules of sperm mitochondria in the mitochondrial sheath surrounding the outer dense fibers and axoneme. To investigate the function of SMCP, we generated mice with a targeted disruption of the gene Smcp by homologous recombination. Homozygous mutant males on a mixed genetic background (C57BL/6J × 129/Sv) are fully fertile, while they are infertile on the 129/Sv background, although spermatogenesis and mating are normal. Homozygous Smcp−/− female mice are fertile on both genetic backgrounds. Electron microscopical examination demonstrated normal structures of sperm head, mitochondria, and tail. In vivo experiments with sperm of Smcp−/− 129/Sv mice revealed that the migration of spermatozoa from the uterus into the oviduct is reduced. This result is supported by the observation that sperm motility as determined by the computer-assisted semen analysis system (CASA) is significantly affected as compared to wild-type spermatozoa. In vitro fertilization assays showed that Smcp-deficient spermatozoa are able to bind to the oocyte but that the number of fertilized eggs is reduced by more than threefold relative to the wild-type control. However, removal of the zona pellucida resulted in an unaffected sperm-egg fusion which was monitored by the presence of pronuclei and generation of blastocyts. These results indicate that the infertility of the male Smcp−/− mice on the 129/Sv background is due to reduced motility of the spermatozoa and decreased capability of the spermatozoa to penetrate oocytes.

Using videotaped specimens to test quality control in a computer-assisted semen analysis system

2000 ◽  
Vol 73 (3) ◽  
pp. 636-640 ◽  
Author(s):  
William R. Boone ◽  
Jeffrey M. Jones ◽  
Sander S. Shapiro
Keyword(s):  
Quality Control ◽  
Semen Analysis ◽  
Computer Assisted ◽  
Test Quality ◽  
Analysis System

scholarly journals A Review on Animals Semen Characteristics: Fertility, Reproduction and Development

2019 ◽  
pp. 1-9
Author(s):  
Farzad Moradpour
Keyword(s):  
Structural Integrity ◽  
Current Knowledge ◽  
Semen Analysis ◽  
Kinetic Property ◽  
Genetic Material ◽  
Computer Assisted ◽  
Semen Characteristics ◽  
The Individual

In this research, the goal of review was summarizing the current knowledge of the methods available to assess in vitro quality of frozen-thawed bovine spermatozoa also, a review on animal’s semen characteristics: fertility, reproduction and development after AI with that semen. Artificial insemination (AI) is the first generation reproductive biotechnology that has made a deep contribution to the genetics improvement in several animals. A fertile ejaculate must meet certain semen characteristics quality standards, such as: normal morphology, active energy metabolism, progressive motility, structural integrity and functionality of the membrane, penetration capacity and optimum transfer of genetic material. The percentage of total motile spermatozoa in normal canine ejaculates is between 70 to 90%. By the way, there are a lot of parameters that able to change on the composition and structure of various sperm plasma member domains, such as change temperature and sensitive to any theirs environments in vivo and vitro (tropical climates), season also nutrition. Computer-assisted semen analysis (CASA) is primarily used to obtain accurate and objective kinetic sperm measurements that gives extensive information about the kinetic property of the ejaculate based on measurements of the individual sperm cells.

scholarly journals Two Protocols of Cryopreservation of Goat Semen with the Use of Computer-Assisted Semen Analysis System

2007 ◽  
Vol 76 (4) ◽  
pp. 601-604 ◽  
Author(s):  
R. Kozdrowski ◽  
A. Dubiel ◽  
W. Bielas ◽  
M. Dzięcioł
Keyword(s):  
Citric Acid ◽  
Semen Analysis ◽  
Egg Yolk ◽  
Computer Assisted ◽  
Semen Cryopreservation ◽  
Velocity Amplitude ◽  
Head Displacement ◽  
Analysis System ◽  
Thawing Procedure ◽  
Lateral Head

The objective of the study was a comparison of two protocols of goat semen cryopreservation with the use of computer-assisted semen analysis system. Twenty ejaculates obtained with electroejaculation method were assessed. Each ejaculate was divided in half and frozen according to two protocols. In protocol I semen was centrifuged in order to remove its plasma and diluted in Tris buffer extender containing glucose, citric acid and glycerol with 20% addition of egg yolk. Protocol II did not include removal of plasma and the extender contained 1.5% egg yolk. It was shown that the removal of semen plasma improved motility of goat spermatozoa following freezing/thawing with respect to the following motility indicators: motility, average path velocity, amplitude of lateral head displacement at p < 0.05, and straight velocity, straightness and linearity at p < 0.01. In conclusion, the removal of semen plasma through centrifugation improved motility properties of goat semen following the freezing/thawing procedure.

scholarly journals Real-Time Single Cell Monitoring: Measurement and Counting of Motile Sperm Using LCR Impedance-Integrated Microfluidic Device

Micromachines ◽  
2019 ◽  
Vol 10 (10) ◽  
pp. 647 ◽  
Author(s):  
Chalinee Phiphattanaphiphop ◽  
Komgrit Leksakul ◽  
Rungrueang Phatthanakun ◽  
Apirak Suthummapiwat
Keyword(s):  
Single Cell ◽  
Real Time ◽  
Low Cost ◽  
Semen Analysis ◽  
Sperm Concentration ◽  
Thin Metal Film ◽  
Computer Assisted ◽  
Cell Detection ◽  
Motile Sperm ◽  
Single Cell Detection

In this research, we aimed to count the ratio of the number of motile to immotile sperm for patients with infertility problems based on a low-sperm-concentration examination. The microfluidic system consists of two series of applications: The conventional separation of motile sperm and the proposed inductance (L), capacitance (C), and resistance (R) or LCR impedance sperm counter. In the experiment, 96% of motile sperm were isolated from nonmotile sperm in the first part and transported to the second part to count and calculate real-time sperm concentration. A pair of microelectrodes composed of thin metal film were integrated between microchannels, resulting in a peak signal for LCR single-cell detection, as well as the estimated total sperm concentration. A minimum of 10 µL of the sperm sample was completely analyzed with an accuracy of 94.8% compared with the standard computer-assisted semen analysis (CASA) method. This method could be applied for low-cost sperm separation and counting in the future.

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