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The effects of cadmium and diazinon administration on rat sperm motility parameters were performed using a Computer Assisted Semen Analyzer (CASA) system. CASA was made on semen samples collected 36 h following an intraperitoneal injection of physiological solution (control group), diazinon (20 mg/kg body wt), cadmium (2 mg/kg body wt), separately and in combination. Sperm kinematic measurements revealed significant decline in the percentage of motile spermatozoa (P

Abstract Background Different parameters are assessed as part of the semen analysis but a standard protocol for evaluation of goat semen is still missing. The aim of this study was to analyse two different factors affecting buck sperm quality in the post-collection period prior to adding the extender. Here we examined the effects of two handling temperatures (20 °C, 37 °C) and various examination time points (3–30 min) after semen collection. Results Examination time point had a significant influence on raw sperm viability (p 0.05), motility (p > 0.05), with the exception of fast moving sperm (p = 0.04), or on semen pH (p > 0.05). Conclusion Examination time point was identified as factor strongly influencing raw peacock buck semen after collection. Raw goat semen can tolerate room temperatures for at least 10 min without impacting overall semen quality. In order to obtain comparable results, semen samples should always be examined within 10 min after collection.

The effect of Alpha-lipoic acid (ALA) on lipid peroxidation, catalase activity and post-thaw semen quality of Achai bulls (Bos indicus) was studied. Semen samples (n=28), collected from adult Achai bulls (n=4) at weekly intervals, were diluted with Tris-citric acid egg yolk extender having various concentrations of ALA (0.00, 0.25, 0.50, 1.00 and 2.00mM) and frozen in liquid nitrogen (-196ºC). Post-thaw semen assessment revealed that the lipid peroxidation decreased, while catalase activity increased (P<0.05) in all extenders containing ALA as compared to control, except in extender containing 2.00mM in which catalase activity decreased (P > 0.05) than control. Moreover, post-thaw semen quality parameters i.e., sperm viability and plasma membrane integrity were higher (P<0.05) in extender having 0.25 and 0.50 mM ALA compared to control, whereas mitochondrial membrane potential was significantly improved only with 0.50mM ALA as compared to control and all other ALA concentrations used. However, acrosome and DNA integrity of spermatozoa increased in extender supplemented with 0.25, 0.50 and 1.00mM ALA (P<0.05) compared to control. Computer Assisted Semen Analysis (CASA) of post-thaw semen revealed that total motility (%) and curvilinear velocity (µm/s), progressive motility (%), progressive fast motility (%) and beat cross frequency (Hz), and straight-line velocity (µm/s) and average path velocity (µm/s) were higher (P < 0.05) for 0.50 and 1.00mM, 0.25 and 0.50mM and 0.50 and 2.00mM ALA concentrations, respectively, than control. In conclusion, the addition of 0.50mM ALA in Tris-citric acid, fructose, egg yolk glycerol extender can perform optimum among all ALA concentrations (0.00, 0.25, 1.00 and 2.00mM) used, in terms of minimizing the oxidative stress and improving the post-thaw semen quality of Achai bulls. [ABSTRACT FROM AUTHOR]

Abstract We analyzed seminal plasma composition of 27 shortnose sturgeon (Acipenser brevirostrum) from both captive and wild populations. Osmolality, pH, sodium, potassium, chloride, and calcium were measured as possible predictors of spermatozoa swimming velocity and percent motility, which are proxies for sperm quality. Principal components analysis resulted in two interpretable components that explained 74% of the variation in the data. The first principal component, PC1, was positively loaded with sodium and potassium, and negatively loaded with chloride. The second principal component, PC2, was positively loaded with calcium and osmolality, and negatively loaded with pH. There were significant differences in seminal plasma composition between wild and captive sourced sperm for both main components (PC1 p -value =.00229, PC2 p-value =.00304), with captive fish having lower sodium, potassium, and osmolality, and higher chloride than wild fish. Cluster analysis of PCA data resulted in three distinct clusters that were characterized by source (p -value =.00458). All subsets regressions showed that seminal plasma sodium content was the best single predictor for swimming velocity and percent motility of both captive and wild sourced sperm. These differences in seminal plasma composition could be contributing to the observed lower quality sperm from captive individuals in this species. Information presented here provides insight on shortnose sturgeon reproduction for both conservation and aquaculture programs. Highlights • Lower motility in spermatozoa from captive shortnose sturgeon compared to wild individuals. • Increased sodium, potassium, and osmolality in seminal plasma of wild shortnose sturgeon. • No significant effect of hormonal treatment on seminal plasma composition or spermatozoa kinematics in Acipenser brevirostrum • Seminal plasma sodium content is best single predictor of sperm kinematics & motility in wild and captive shortnose sturgeon. [ABSTRACT FROM AUTHOR]

Here, we show the effects of sexual steroids, progesterone, testosterone, or estradiol on motility parameters of boar sperm. Sixteen commercial seminal doses, four each of four adult boars, were analyzed using computer assisted sperm analysis (CASA). Mean values of motility parameters were analyzed by bivariate and multivariate statistics. Principal component analysis (PCA), followed by hierarchical clustering, was applied on data of motility parameters, provided automatically as intervals by the CASA system. Effects of sexual steroids were described in the kinematic subpopulations identified from multivariate statistics. Mean values of motility parameters were not significantly changed after addition of sexual steroids. Multivariate graphics showed that sperm subpopulations were not sensitive to the addition of either testosterone or estradiol, but sperm subpopulations responsive to progesterone were found. Distribution of motility parameters were wide in controls but sharpened at distinct concentrations of progesterone. We conclude that kinematic sperm subpopulations responsive to progesterone are present in boar semen, and these subpopulations are masked in evaluations of mean values of motility parameters. © 2017 International Society for Advancement of Cytometry [ABSTRACT FROM AUTHOR]

Here we show the effects of repeated electroejaculation (EE) on mean values of motility, mitochondrial functionality, and expression of active caspases on goat sperm obtained by EE. Evaluations were done using CASA and flow cytometry. A strategy for identification of kinematic sperm subpopulations, when individual data of sperm are not provided by the CASA system, is provided. Fifty semen samples, five of each of ten adult creole goats, were obtained by electroejaculation. Mean values of total motility, progressive motility and flow cytometry evaluations were compared among EEs. Relationships among mean values of variables were investigated using Spearman correlation and principal component analysis (PCA). For identification of kinematic sperm subpopulations, PCA followed by hierarchical clustering was applied on data of the intervals provided automatically by the CASA system. Total motility does no change after repeated EE. Mean values of motility parameters and molecular markers were unrelated in multivariate space, but bivariate correlations were found. Values in upper and lower intervals defined clearly the sperm subpopulations, which had motility parameters changing over time. Taken together, our results show that repeated EE does not affect mean values of total motility, that molecular markers are not related with motility parameters, and that it is possible to identify kinematic sperm subpopulations when individual data, of motility parameters, are not provided by the CASA system. [ABSTRACT FROM AUTHOR]

Tall fescue possesses heat, drought, and pest resistance conferred to the plant by its mutualistic relationship with the ergot alkaloid producing fungal endophyte, Neotyphodium coenophialum. The objec-tive of this study was to evaluate the impact of ergot alkaloid consumption on growth, scrotal circumference (SC), and semen quality. The SC measurement and percentage of motile and normal sperm were used to determine if a bull passed the breeding soundness exam (BSE) requirements. Bulls (n = 14) between 13 and 16 mo of age exhibiting >32 cm SC and having passed a BSE were assigned to 1 of 2 dietary treatments account-ing for BCS and BW. Bulls were fed the treatment diet containing toxic tall fescue seed (E+; 0.8 μg of ergo-valine and ergovalanine/g DM) or the control diet con-taining endophyte-free nontoxic tall fescue seed (E-) for 126 d. Blood samples were collected and BSE and BCS accessed at the start of the test (d 0) and every 21 d to the end of test (d 126). Weights were obtained on d 0 and d 126. Semm prolactin (PRL) concentrations were affected by treatment x day interactions (P = 0.04) veri-fying the effectiveness of the E+ diet. Bulls consuming the E+ diet exhibited declining PRL concentrations from 250 ± 52.1 ng/mL on d 0 to 30.6 ± 46.9 ng/mL by d 126 whereas bulls receiving the E- ration maintained serum PRL concentrations greater than or equal to 226.7 ± 50.4 ng/mL across the 126-d study. Body condition score (P = 0.4) and BW (P = 0.4) were not different between treatments. No difference due to treatment was observed for the percentage of bulls passing a standard BSE exam (P = 0.6) and no treatment effect was observed for any semen characteristic measured by computer-assisted semen analysis (CASA; P > 0.2). The SC was nega-tively affected by treatment x day interaction (P = 0.04) with E- bulls exhibiting a larger SC at d 126 compared with E+ bulls of 36.7 ± 0.8 versus 34.3 ± 0.8 cm, respec-tively. Within treatment, E+ bulls exhibited a decrease in SC (P = 0.0001) with ad 0 SC of 37.3 ± 0.8 cm and dropping to 34.3 ± 0.8 by d 126. Theoretically, reduced SC would negatively impact semen quality, but this was not observed. However, CASA and BSE evaluation data are consistent with recent reports indicating that bulls grazing E+ tall fescue exhibited only subtle, if any, dif-ferences on semen characteristics. [ABSTRACT FROM AUTHOR]

In order to identify unreliable data in a dataset of motility parameters obtained from a pilot study acquired by a veterinarian with experience in boar semen handling, but without experience in the operation of a computer assisted sperm analysis (CASA) system, a multivariate graphical and statistical analysis was performed. Sixteen boar semen samples were aliquoted then incubated with varying concentrations of progesterone from 0 to 3.33 μg/ml and analyzed in a CASA system. After standardization of the data, Chernoff faces were pictured for each measurement, and a principal component analysis (PCA) was used to reduce the dimensionality and preprocess the data before hierarchical clustering. The first twelve individual measurements showed abnormal features when Chernoff faces were drawn. PCA revealed that principal components 1 and 2 explained 63.08% of the variance in the dataset. Values of principal components for each individual measure- ment of semen samples were mapped to identify differences among treatment or among boars. Twelve individual measure ments presented low values of principal component 1. Confidence ellipses on the map of principal components showed no statistically significant effects for treatment or boar. Hierarchical clustering realized on two first principal components produced three clusters. Cluster 1 contained evaluations of the two first samples in each treatment, each one of a different boar. With the exception of one individual measurement, all other measurements in cluster 1 were the same as observed in abnormal Chernoff faces. Unreliable data in cluster 1 are probably related to the operator inexperience with a CASA system. These findings could be used to objectively evaluate the skill level of an operator of a CASA system. This may be particularly useful in the quality control of semen analysis using CASA systems. [ABSTRACT FROM AUTHOR]

In this study the effect of six activating buffers on the movement parameters of sperm were determined and short-term storage of semen in TLP buffer was attempted (0.292 g NaCl; 0.012g KCl; 0.011g CaCl2; 0.004g MgCl2; 0.105g NaHCO3; 0.002g NaH2PO4; 50 mL; pH 8.6). Sperm was collected from five orfe individual, and spermiation was stimulated by means of an intraperitoneal Ovopel injection. The basic parameters of spermatozoa motility were measured after the semen was diluted with six different activating solutions, previously used successfully in other fish species. The motility analysis was conducted on a Crismas apparatus. Additionally, short-term storage of semen in TLP buffer was attempted. Subsequently, motility parameters were verified after 0 (Control), 24 and 120 h of storage at 4°C. It has been found that Lahnsteiner's buffer (100 mM NaCl, 10 mM Tris, 0.5% albumin, 199 mOsmkg-1) was found to be the most effective in sperm activation. In this paper, the spermatozoa motility of colourful ide is indicated for the first time. Finally, there was a successful attempt at short-term sperm storage for five days. For artificial insemination, it is very important to select the most effective solution to stimulate sperm motility. Data regarding sperm manipulation of orfe are scarce, so the aim of the study was to determine the basic sperm quality parameters of the colour ide form, i.e. Leuciscus idus aberr orfus. [ABSTRACT FROM AUTHOR]

The effects of cadmium and diazinon administration on rat sperm motility parameters were performed using a Computer Assisted Semen Analyzer (CASA) system. CASA was made on semen samples collected 36 h following an intraperitoneal injection of physiological solution (control group), diazinon (20 mg/kg body wt), cadmium (2 mg/kg body wt), separately and in combination. Sperm kinematic measurements revealed significant decline in the percentage of motile spermatozoa (P<0.05), significant higher amplitude of lateral head displacement (ALH) (P<0.01) with significant decline in beat cross frequency (BCF) (P<0.001) after the diazinon administration. Cadmium-treated males exhibited significantly lower percentage of motile spermatozoa (P<0.01) and percentage of spermatozoa with progressive motility (P<0.001) with significant decrease in straightness (STR) and BCF (P<0.05). Simultaneous exposure to cadmium and diazinon led to significant increase in ALH (P<0.05) and significant decrease in BCF (P<0.001). Cadmium and diazinon exerted deleterious effect inducing spermatozoa motility alterations which could be subsequently negatively related to male fertility. [ABSTRACT FROM AUTHOR]

Abstract: Computer-assisted semen analysis (CASA) is primarily used to obtain accurate and objective kinetic sperm measurements. Additionally, AI centers use computer-assessed sperm concentration in the sample as a basis for calculating the number of insemination doses available from a given ejaculate. The reliability of data is often limited and results can vary even when the same CASA systems with identical settings are used. The objective of the present study was to develop a computer-based training module for standardized measurements with a CASA system and to evaluate its training effect on the quality of the assessment of sperm motility and concentration. A digital versatile disc (DVD) has been produced showing the standardization of sample preparation and analysis with the CASA system SpermVision™ version 3.0 (Minitube, Verona, WI, USA) in words, pictures, and videos, as well as the most probable sources of error. Eight test persons educated in spermatology, but with different levels of experience with the CASA system, prepared and assessed 10 aliquots from one prediluted bull ejaculate using the same CASA system and laboratory equipment before and after electronic learning (e-learning). After using the e-learning application, the coefficient of variation was reduced on average for the sperm concentration from 26.1% to 11.3% (P ≤ 0.01), and for motility from 5.8% to 3.1% (P ≤ 0.05). For five test persons, the difference in the coefficient of variation before and after use of the e-learning application was significant (P ≤ 0.05). Individual deviations of means from the group mean before e-learning were reduced compared with individual deviations from the group mean after e-learning. According to a survey, the e-learning application was highly accepted by users. In conclusion, e-learning presents an effective, efficient, and accepted tool for improvement of the precision of CASA measurements. This study provides a model for the standardization of other laboratory procedures using e-learning. [Copyright &y& Elsevier]

Objective: To evaluate the in vitro effect of benzo[a]pyrene on sperm hyperactivation and acrosome status in normozoospermic semen samples of nonsmokers analyzed by computer-assisted semen analysis (CASA). Design: Experimental in vitro study. Setting: Andrology laboratory. Patient(s): Thirteen proven fertile, normozoospermic, and nonsmoking men. Intervention(s): Spermatozoa were washed free of seminal plasma and were treated with different concentrations of benzo[a]pyrene and compared with controls treated with medium alone. The benzo[a]pyrene concentrations were: 100, 50, 25, and 12.5 μg/mL. Main Outcome Measure(s): Effect of varying concentrations of benzo[a]pyrene on sperm hyperactivation and acrosomal reaction. Result(s): A statistically significant increase in sperm hyperactivation was observed in presence of benzo[a]pyrene at concentrations of ≥50 μg/mL. The result of the acrosome halo test showed that concentrations of benzo[a]pyrene ≥25 μg/mL statistically significantly decreased the percentage of halo formation, indicating an inappropriate (false) acrosome reaction. Conclusion(s): Benzo[a]pyrene statistically significantly affected sperm functional competence as evidenced by increased hyperactivation as well as premature acrosomal reaction. [Copyright &y& Elsevier]

Purpose To determine the efficiency of sperm motion characteristics as predictors for normal (2PN) and polypronulceate (PPN) zygotes in IVF. Methods A retrospective cohort analysis for a total of 230 couples undergoing IVF treatment in a single infertility center. Result(s) Subsequent to semen analysis and hemizona assay, unexpected fertilization failure would appear to have occurred only extremely rarely (1/236, 0.4%). The rate of PPN, however, did arise and appeared to be related to certain sperm motion characteristics, such as lateral head displacement and concentration of progressive motile sperm. Interestingly, the patients featuring a high PPN rate (>20%) was associated with a greater pregnancy rate than those featuring a low PPN rate (<20%). Conclusion The sperm motion characteristics examined herein could be utilized to predict the rate of PPN in IVF. In order to enhance the rate of 2PN and maintain the relative high rate of clinical pregnancy, an efficient method needs further investigation and development. [ABSTRACT FROM AUTHOR]

We purified two proteins with molecular masses of approximately 50 kDa and 80 kDa with N-terminal sequences similar to those of alpha1-antitrypsin (a1AT) and transferrin indicating that they are identical to or highly homologous to these proteins. Proteins from human follicular fluid were purified after ammonium sulfate fractionation followed by water dialysis and High Performance Liquid Chromatography. The fraction of peak 3 showed a single band on electrophoresis and its N-terminal amino acid sequence was similar to that of human serum transferrin. The fraction of peak 10 proved to be a glycoprotein and its N-terminal amino acid sequence was similar to that of human serum a1AT. There are indications that transferrin may be involved in the fertilization process. Sperm motion was assessed employing computer-assisted semen analysis. The addition of purified protein to prepared sperm samples from normospermic men significantly increases the straight-line velocity (VSL), the amplitude of lateral head displacement (ALH) and the number of progressively motile sperm. a1AT does not seem to have a stimulatory effect on sperm motility. [Copyright &y& Elsevier]

Excessive alcohol consumption has been associated with impaired reproductive function by causing the inhibition of penile tumescence and ejaculatory capability. Alcohol intoxication has also been implicated in impaired spermatogenesis and an increase in sperm structural anomalies. The aim of this study was to determine the direct effects of alcohol on sperm motility and morphology in vitro. Semen samples from 67 subjects were prepared using density centrifugation. Ethanol was added, at concentrations in serum equivalent to social, moderate and heavy drinking, to the medium in which the spermatozoa were cultured. Sperm motility was assessed using computer assisted semen analysis and morphology was assessed by Tygerberg strict criteria after 0, 15, 30, 60, 120, 180 and 240 min exposure. Each concentration of ethanol produced significant decreases in the percentage progressive motility, straight line velocity and curvilinear velocity. The amplitude of lateral head displacement was also depressed by 300 and 500 mg dL-1 of ethanol. A significant decrease in the number of spermatozoa with normal morphology and an increase in irreversible tail defects were observed after exposure to 300 mg dL-1 ethanol. When alcohol is added directly to sperm, at concentrations equivalent to those in serum after moderate and heavy drinking, damaging effects are observed in both sperm motility and morphology. [ABSTRACT FROM AUTHOR]

Two hundred and seven patients with male infertility were investigated. Total sperm concentration and percent progressive motility by SQA IIB showed high correlations with those of conventional manual method. Percent of normal morphology showed a significant correlation among these techniques. The sperm motility index (SMI) and total functional sperm concentration (TFSC) demonstrated high correlations with any variables of manual analysis. Only velocity and amplitude of lateral head displacement showed significant correlations with the variables obtained by SQA IIB, especially with SMI and TFSC. It was suggested that SQA IIB could be a useful instrument in the clinical practice of infertility as a screening test for semen quality. [ABSTRACT FROM AUTHOR]

Abstract: In order to obtain a more standardised semen motility evaluation, Varkens KI Nederland has introduced a computer assisted semen analysis (CASA) system in all their pig AI laboratories. The repeatability of CASA was enhanced by standardising for: 1) an optimal sample temperature (39 °C); 2) an optimal dilution factor; 3) optimal mixing of semen and dilution buffer by using mechanical mixing; 4) the slide chamber depth, and together with the previous points; 5) the optimal training of technicians working with the CASA system; and 6) the use of a standard operating procedure (SOP). Once laboratory technicians were trained in using this SOP, they achieved a coefficient of variation of < 5% which was superior to the variation found when the SOP was not strictly used. Microscopic semen motility assessments by eye were subjective and not comparable to the data obtained by standardised CASA. CASA results are preferable as accurate continuous motility dates are generated rather than discrimination motility percentage increments of 10% motility as with motility estimation by laboratory technicians. The higher variability of sperm motility found with CASA and the continuous motility values allow better analysis of the relationship between semen motility characteristics and fertilising capacity. The benefits of standardised CASA for AI is discussed both with respect to estimate the correct dilution factor of the ejaculate for the production of artificial insemination (AI) doses (critical for reducing the number of sperm per AI doses) and thus to get more reliable fertility data from these AI doses in return. [Copyright &y& Elsevier]